doc_id sent_index relation_id relation trigger trigger_offset arg_num arg_head arg_protein arg_domain arg_site arg_sugar arg_head_offset arg_base_np arg_base_np_offset arg_np arg_np_offset sent_text PSource NProtein NID SiteSource SiteName 29048990 3 0 gly glycosylation 604:616 arg1 strains 640:646 influenza strains 630:646 influenza strains 630:646 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 7 1 theme HCD/CID-MS/MS 1121:1133 arg1 spectrometry 1147:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 9 2 theme stalk 1467:1471 arg1 region 1473:1478 the stalk region 1463:1478 the stalk region (N28, N40, N304 and N498) 1463:1504 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 2 3 theme strain 516:521 arg1 properties 498:507 the antigenic properties 484:507 the antigenic properties of the strain 484:521 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 2 3 theme strain 516:521 arg1 range 449:453 a wide range 442:453 a wide range of biological activities 442:478 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 3 4 theme vaccines 701:708 arg1 development 677:687 the development 673:687 the development of improved vaccines 673:708 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 7 5 theme A/New 1247:1251 arg1 Caledonia/20/1999 1253:1269 A/New Caledonia/20/1999 1247:1269 A/New Caledonia/20/1999 1247:1269 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 7 5 theme A/New 1247:1251 arg1 strain 1239:1244 an egg-grown H1N1 reference strain 1211:1244 an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1211:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 1 6 theme glycosylation 190:202 arg1 neuraminidase 276:288 neuraminidase 276:288 neuraminidase (NA) 276:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 6 theme glycosylation 190:202 arg1 hemagglutinin 253:265 hemagglutinin 253:265 hemagglutinin (HA) 253:270 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 6 theme glycosylation 190:202 arg1 alterations 212:222 glycosylation profile alterations 190:222 glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA) 190:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 9 7 dep region 1473:1478 arg1 N498 1500:1503 N498 1500:1503 N498 1500:1503 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 7 dep region 1473:1478 arg1 N304 1491:1494 N304 1491:1494 N304 1491:1494 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 7 dep region 1473:1478 arg1 N40 1486:1488 N40 1486:1488 N40 1486:1488 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 11 8 theme influenza 1935:1943 arg1 vaccines 1945:1952 influenza vaccines 1935:1952 influenza vaccines 1935:1952 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 1 9 theme alterations 212:222 arg1 role 182:185 a significant role 168:185 a significant role 168:185 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 11 10 theme other 1777:1781 arg1 viruses 1783:1789 other viruses 1777:1789 other viruses 1777:1789 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 1 11 from glycoproteins 237:249 arg1 role 182:185 a significant role 168:185 a significant role 168:185 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 3 12 theme key 539:541 arg1 role 543:546 the key role 535:546 the key role of glycans in determining antigenicity 535:585 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 8 13 theme glycosylation 1318:1330 arg1 sites 1332:1336 glycosylation sites 1318:1336 glycosylation sites identified by primary sequence analysis 1318:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 5 14 theme potential 916:924 arg1 sites 940:944 potential glycosylation sites 916:944 potential glycosylation sites 916:944 Nonetheless, this sequence-based approach can only predict potential glycosylation sites. 29048990 6 15 theme predicted 1026:1034 arg1 sites 1050:1054 predicted glycosylation sites 1026:1054 predicted glycosylation sites 1026:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 2 16 theme biological 458:467 arg1 activities 469:478 biological activities 458:478 biological activities 458:478 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 7 17 theme strain 1239:1244 arg1 HA 1205:1206 HA 1205:1206 HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1205:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 6 18 theme occupation 1012:1021 arg1 confirmation 992:1003 experimental confirmation 979:1003 experimental confirmation of the occupation of predicted glycosylation sites 979:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 8 19 theme primary 1352:1358 arg1 analysis 1369:1376 primary sequence analysis 1352:1376 primary sequence analysis 1352:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 7 20 theme H1N1 1224:1227 arg1 Caledonia/20/1999 1253:1269 A/New Caledonia/20/1999 1247:1269 A/New Caledonia/20/1999 1247:1269 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 7 20 theme H1N1 1224:1227 arg1 strain 1239:1244 an egg-grown H1N1 reference strain 1211:1244 an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1211:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 1 21 from humans 148:153 arg1 study 110:114 The study 106:114 The study of influenza virus evolution in humans 106:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 2 22 theme antigenic 488:496 arg1 properties 498:507 the antigenic properties 484:507 the antigenic properties of the strain 484:521 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 4 23 theme viral 738:742 arg1 RNA 744:746 viral RNA 738:746 viral RNA 738:746 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 0 24 theme mass 87:90 arg1 spectrometry 92:103 tandem mass spectrometry 80:103 tandem mass spectrometry 80:103 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 1 25 theme seasonal 321:328 arg1 strains 343:349 both seasonal and pandemic strains 316:349 both seasonal and pandemic strains 316:349 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 26 from evolution 135:143 arg1 humans 148:153 humans 148:153 humans 148:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 6 27 theme experimental 979:990 arg1 confirmation 992:1003 experimental confirmation 979:1003 experimental confirmation of the occupation of predicted glycosylation sites 979:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 10 28 theme broad 1615:1619 arg1 microheterogeneity 1628:1645 a broad glycan microheterogeneity 1613:1645 a broad glycan microheterogeneity 1613:1645 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 7 29 theme HA 1205:1206 arg1 profile 1194:1200 the site-specific profile 1176:1200 the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1176:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 1 30 theme pandemic 334:341 arg1 strains 343:349 both seasonal and pandemic strains 316:349 both seasonal and pandemic strains 316:349 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 2 31 theme glycosylation 412:424 arg1 sites 426:430 glycosylation sites 412:430 glycosylation sites 412:430 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 1 32 theme virus 129:133 arg1 evolution 135:143 influenza virus evolution 119:143 influenza virus evolution in humans 119:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 2 33 theme Viral 352:356 arg1 drift 368:372 Viral antigenic drift 352:372 Viral antigenic drift 352:372 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 0 34 theme Site-specific 0:12 arg1 profile 28:34 Site-specific glycosylation profile 0:34 Site-specific glycosylation profile of influenza A (H1N1) 0:56 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 1 35 from study 110:114 arg1 humans 148:153 humans 148:153 humans 148:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 11 36 theme vaccines 1945:1952 arg1 improvement 1920:1930 the improvement 1916:1930 the improvement of influenza vaccines 1916:1952 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 8 37 theme glycan 1414:1419 arg1 structures 1421:1430 glycan structures 1414:1430 glycan structures 1414:1430 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 4 38 gly glycosite 792:800 arg2 glycosite 792:800 glycosite modifications 792:814 glycosite modifications 792:814 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 3 39 theme glycosylation 604:616 arg1 profiles 618:625 the glycosylation profiles 600:625 the glycosylation profiles of influenza strains 600:646 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 11 40 theme biological 1886:1895 arg1 activities 1897:1906 biological activities 1886:1906 biological activities 1886:1906 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 0 41 theme influenza 39:47 arg1 A 49:49 influenza A 39:49 influenza A (H1N1) 39:56 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 0 41 theme influenza 39:47 arg1 H1N1 52:55 H1N1 52:55 H1N1 52:55 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 10 42 gly glycosite 1711:1719 arg2 glycosite 1711:1719 each glycosite 1706:1719 each glycosite 1706:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 0 43 gly glycosylation 14:26 arg1 A 49:49 influenza A 39:49 influenza A (H1N1) 39:56 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 0 43 gly glycosylation 14:26 arg1 H1N1 52:55 H1N1 52:55 H1N1 52:55 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 6 44 theme sites 1050:1054 arg1 occupation 1012:1021 the occupation 1008:1021 the occupation of predicted glycosylation sites 1008:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 4 45 theme glycosite 792:800 arg1 modifications 802:814 glycosite modifications 792:814 glycosite modifications 792:814 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 7 46 theme mass 1142:1145 arg1 spectrometry 1147:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 9 47 theme protein 1570:1576 arg1 head 1531:1534 the globular head 1518:1534 the globular head (N71, N104, N142 and N177) of the protein 1518:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 48 used occupied 1582:1589 arg2 head 1531:1534 the globular head 1518:1534 the globular head (N71, N104, N142 and N177) of the protein 1518:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 48 used occupied 1582:1589 arg2 region 1473:1478 the stalk region 1463:1478 the stalk region (N28, N40, N304 and N498) 1463:1504 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 48 used occupied 1582:1589 arg2 four 1510:1513 four 1510:1513 four 1510:1513 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 48 used occupied 1582:1589 arg2 sequons 1452:1458 Four glycosylation sequons 1433:1458 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein 1433:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 3 49 theme strains 640:646 arg1 profiles 618:625 the glycosylation profiles 600:625 the glycosylation profiles of influenza strains 600:646 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 4 50 theme great 761:765 arg1 insight 767:773 great insight 761:773 great insight into the role of glycosite modifications in altering virulence and pathogenicity 761:854 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 11 51 theme different 1805:1813 arg1 strains 1825:1831 different influenza strains 1805:1831 different influenza strains 1805:1831 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 2 52 gly glycosylation 412:424 arg2 sites 426:430 glycosylation sites 412:430 glycosylation sites 412:430 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 6 53 gly glycosylation 1036:1048 arg2 sites 1050:1054 predicted glycosylation sites 1026:1054 predicted glycosylation sites 1026:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 10 54 theme present 1695:1701 arg1 compositions 1682:1693 glycan compositions 1675:1693 glycan compositions present on each glycosite 1675:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 1 55 theme viral 231:235 arg1 glycoproteins 237:249 the viral glycoproteins 227:249 the viral glycoproteins 227:249 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 7 56 theme egg-grown 1214:1222 arg1 Caledonia/20/1999 1253:1269 A/New Caledonia/20/1999 1247:1269 A/New Caledonia/20/1999 1247:1269 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 7 56 theme egg-grown 1214:1222 arg1 strain 1239:1244 an egg-grown H1N1 reference strain 1211:1244 an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1211:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 10 57 theme glycan 1675:1680 arg1 compositions 1682:1693 glycan compositions 1675:1693 glycan compositions present on each glycosite 1675:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 7 58 used utilized 1112:1119 arg2 we 1109:1110 we 1109:1110 we 1109:1110 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 6 59 theme few 1088:1090 arg1 strains 1092:1098 a few strains 1086:1098 a few strains 1086:1098 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 2 60 dep number 389:394 arg1 the 385:387 the 385:387 the 385:387 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 9 61 theme globular 1522:1529 arg1 head 1531:1534 the globular head 1518:1534 the globular head (N71, N104, N142 and N177) of the protein 1518:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 3 62 theme improved 692:699 arg1 vaccines 701:708 improved vaccines 692:708 improved vaccines 692:708 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 4 63 theme modifications 802:814 arg1 role 784:787 the role 780:787 the role of glycosite modifications in altering virulence and pathogenicity 780:854 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 1 64 theme significant 170:180 arg1 role 182:185 a significant role 168:185 a significant role 168:185 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 6 65 theme experimental 954:965 arg1 challenges 967:976 experimental challenges 954:976 experimental challenges 954:976 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 3 66 theme role 543:546 arg1 view 527:530 view 527:530 view of the key role of glycans in determining antigenicity 527:585 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 1 67 theme profile 204:210 arg1 neuraminidase 276:288 neuraminidase 276:288 neuraminidase (NA) 276:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 67 theme profile 204:210 arg1 hemagglutinin 253:265 hemagglutinin 253:265 hemagglutinin (HA) 253:270 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 67 theme profile 204:210 arg1 alterations 212:222 glycosylation profile alterations 190:222 glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA) 190:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 3 68 from requirement 653:663 arg1 view 527:530 view 527:530 view of the key role of glycans in determining antigenicity 527:585 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 1 69 from alterations 212:222 arg1 glycoproteins 237:249 the viral glycoproteins 227:249 the viral glycoproteins 227:249 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 70 theme influenza 119:127 arg1 virus 129:133 influenza virus 119:133 influenza virus evolution in humans 119:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 6 71 theme glycosylation 1036:1048 arg1 sites 1050:1054 predicted glycosylation sites 1026:1054 predicted glycosylation sites 1026:1054 Due to experimental challenges, experimental confirmation of the occupation of predicted glycosylation sites has only been carried out for a few strains. 29048990 11 72 from impact 1859:1864 arg1 activities 1897:1906 biological activities 1886:1906 biological activities 1886:1906 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 7 73 theme reference 1229:1237 arg1 Caledonia/20/1999 1253:1269 A/New Caledonia/20/1999 1247:1269 A/New Caledonia/20/1999 1247:1269 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 7 73 theme reference 1229:1237 arg1 strain 1239:1244 an egg-grown H1N1 reference strain 1211:1244 an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1211:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 3 74 theme glycans 551:557 arg1 role 543:546 the key role 535:546 the key role of glycans in determining antigenicity 535:585 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 1 75 from role 182:185 arg1 glycoproteins 237:249 the viral glycoproteins 227:249 the viral glycoproteins 227:249 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 75 from role 182:185 arg1 emergence 303:311 the emergence 299:311 the emergence of both seasonal and pandemic strains 299:349 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 76 theme evolution 135:143 arg1 study 110:114 The study 106:114 The study of influenza virus evolution in humans 106:153 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 8 77 theme sites 1332:1336 arg1 occupancy 1305:1313 experimentally the occupancy 1286:1313 experimentally the occupancy of glycosylation sites identified by primary sequence analysis 1286:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 0 78 theme tandem 80:85 arg1 spectrometry 92:103 tandem mass spectrometry 80:103 tandem mass spectrometry 80:103 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 8 79 gly occupancy 1305:1313 arg2 sites 1332:1336 glycosylation sites 1318:1336 glycosylation sites identified by primary sequence analysis 1318:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 5 80 theme sequence-based 875:888 arg1 approach 890:897 this sequence-based approach 870:897 this sequence-based approach 870:897 Nonetheless, this sequence-based approach can only predict potential glycosylation sites. 29048990 2 81 theme wide 444:447 arg1 range 449:453 a wide range 442:453 a wide range of biological activities 442:478 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 8 82 gly glycosylation 1318:1330 arg2 sites 1332:1336 glycosylation sites 1318:1336 glycosylation sites identified by primary sequence analysis 1318:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 1 83 gly glycoproteins 237:249 arg1 glycoproteins 237:249 the viral glycoproteins 227:249 the viral glycoproteins 227:249 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 5 84 gly glycosylation 926:938 arg2 sites 940:944 potential glycosylation sites 916:944 potential glycosylation sites 916:944 Nonetheless, this sequence-based approach can only predict potential glycosylation sites. 29048990 8 85 theme sequence 1360:1367 arg1 analysis 1369:1376 primary sequence analysis 1352:1376 primary sequence analysis 1352:1376 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 11 86 theme influenza 1815:1823 arg1 strains 1825:1831 different influenza strains 1805:1831 different influenza strains 1805:1831 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 2 87 theme activities 469:478 arg1 properties 498:507 the antigenic properties 484:507 the antigenic properties of the strain 484:521 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 2 87 theme activities 469:478 arg1 range 449:453 a wide range 442:453 a wide range of biological activities 442:478 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 4 88 theme RNA 744:746 arg1 analysis 726:733 Sequence-based analysis 711:733 Sequence-based analysis of viral RNA 711:746 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 2 89 theme antigenic 358:366 arg1 drift 368:372 Viral antigenic drift 352:372 Viral antigenic drift 352:372 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 0 90 theme glycosylation 14:26 arg1 profile 28:34 Site-specific glycosylation profile 0:34 Site-specific glycosylation profile of influenza A (H1N1) 0:56 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 1 91 theme strains 343:349 arg1 emergence 303:311 the emergence 299:311 the emergence of both seasonal and pandemic strains 299:349 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 9 92 gly glycosylation 1438:1450 arg2 sequons 1452:1458 Four glycosylation sequons 1433:1458 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein 1433:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 5 93 theme glycosylation 926:938 arg1 sites 940:944 potential glycosylation sites 916:944 potential glycosylation sites 916:944 Nonetheless, this sequence-based approach can only predict potential glycosylation sites. 29048990 9 94 dep head 1531:1534 arg1 N142 1548:1551 N142 1548:1551 N142 1548:1551 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 94 dep head 1531:1534 arg1 N104 1542:1545 N104 1542:1545 N104 1542:1545 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 94 dep head 1531:1534 arg1 N177 1557:1560 N177 1557:1560 N177 1557:1560 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 10 95 attach present 1695:1701 arg1 glycosite 1711:1719 each glycosite 1706:1719 each glycosite 1706:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 10 95 attach present 1695:1701 arg2 compositions 1682:1693 glycan compositions 1675:1693 glycan compositions present on each glycosite 1675:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 4 96 theme Sequence-based 711:724 arg1 analysis 726:733 Sequence-based analysis 711:733 Sequence-based analysis of viral RNA 711:746 Sequence-based analysis of viral RNA has provided great insight into the role of glycosite modifications in altering virulence and pathogenicity. 29048990 0 97 theme A 49:49 arg1 profile 28:34 Site-specific glycosylation profile 0:34 Site-specific glycosylation profile of influenza A (H1N1) 0:56 Site-specific glycosylation profile of influenza A (H1N1) hemagglutinin through tandem mass spectrometry. 29048990 9 98 from sequons 1452:1458 arg1 region 1473:1478 the stalk region 1463:1478 the stalk region (N28, N40, N304 and N498) 1463:1504 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 98 from sequons 1452:1458 arg1 four 1510:1513 four 1510:1513 four 1510:1513 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 9 98 from sequons 1452:1458 arg1 head 1531:1534 the globular head 1518:1534 the globular head (N71, N104, N142 and N177) of the protein 1518:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 29048990 11 99 theme glycosylation 1869:1881 arg1 impact 1859:1864 the impact 1855:1864 the impact of glycosylation on biological activities 1855:1906 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 8 100 theme structures 1421:1430 arg1 heterogeneity 1397:1409 the heterogeneity 1393:1409 the heterogeneity of glycan structures 1393:1430 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 10 101 theme glycan 1621:1626 arg1 microheterogeneity 1628:1645 a broad glycan microheterogeneity 1613:1645 a broad glycan microheterogeneity 1613:1645 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 7 102 theme tandem 1135:1140 arg1 spectrometry 1147:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 HCD/CID-MS/MS tandem mass spectrometry 1121:1158 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 8 103 gly heterogeneity 1397:1409 arg1 structures 1421:1430 glycan structures 1414:1430 glycan structures 1414:1430 We confirmed experimentally the occupancy of glycosylation sites identified by primary sequence analysis and determined the heterogeneity of glycan structures. 29048990 2 104 theme sites 426:430 arg1 location 400:407 location 400:407 location 400:407 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 2 104 theme sites 426:430 arg1 number 389:394 number 389:394 number 389:394 Viral antigenic drift can modify the number and location of glycosylation sites, altering a wide range of biological activities and the antigenic properties of the strain. 29048990 11 105 theme present 1726:1732 arg1 methodology 1734:1744 The present methodology 1722:1744 The present methodology 1722:1744 The present methodology can be applied to characterize other viruses, particularly different influenza strains, to better understand the impact of glycosylation on biological activities and aid the improvement of influenza vaccines. 29048990 3 106 theme influenza 630:638 arg1 strains 640:646 influenza strains 630:646 influenza strains 630:646 In view of the key role of glycans in determining antigenicity, elucidating the glycosylation profiles of influenza strains is a requirement towards the development of improved vaccines. 29048990 10 107 theme great 1656:1660 arg1 diversity 1662:1670 a great diversity 1654:1670 a great diversity of glycan compositions present on each glycosite 1654:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 1 108 dep alterations 212:222 arg1 neuraminidase 276:288 neuraminidase 276:288 neuraminidase (NA) 276:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 108 dep alterations 212:222 arg1 hemagglutinin 253:265 hemagglutinin 253:265 hemagglutinin (HA) 253:270 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 108 dep alterations 212:222 arg1 alterations 212:222 glycosylation profile alterations 190:222 glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA) 190:293 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 108 dep alterations 212:222 arg1 NA 291:292 NA 291:292 NA 291:292 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 1 108 dep alterations 212:222 arg1 HA 268:269 HA 268:269 HA 268:269 The study of influenza virus evolution in humans has revealed a significant role of glycosylation profile alterations in the viral glycoproteins - hemagglutinin (HA) and neuraminidase (NA), in the emergence of both seasonal and pandemic strains. 29048990 10 109 theme compositions 1682:1693 arg1 diversity 1662:1670 a great diversity 1654:1670 a great diversity of glycan compositions present on each glycosite 1654:1719 Our results revealed a broad glycan microheterogeneity, i.e., a great diversity of glycan compositions present on each glycosite. 29048990 7 110 theme site-specific 1180:1192 arg1 profile 1194:1200 the site-specific profile 1176:1200 the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999) 1176:1270 Herein, we utilized HCD/CID-MS/MS tandem mass spectrometry to characterize the site-specific profile of HA of an egg-grown H1N1 reference strain (A/New Caledonia/20/1999). 29048990 9 111 theme glycosylation 1438:1450 arg1 sequons 1452:1458 Four glycosylation sequons 1433:1458 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein 1433:1576 Four glycosylation sequons on the stalk region (N28, N40, N304 and N498) and four on the globular head (N71, N104, N142 and N177) of the protein are occupied. 25450502 0 0 theme recombinant 108:118 arg1 osteopontin 120:130 recombinant osteopontin 108:130 recombinant osteopontin 108:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 5 1 theme sialyl-Le 976:984 arg1 antigens 991:998 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 6 2 gly O-glycopeptides 1094:1108 arg2 O-glycopeptides 1094:1108 the generated O-glycopeptides 1080:1108 the generated O-glycopeptides 1080:1108 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 1 3 theme multiple 171:178 arg1 processes 211:219 multiple physiological and pathological processes 171:219 multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis 171:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 3 theme multiple 171:178 arg1 mineralization 250:263 mineralization 250:263 mineralization 250:263 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 3 theme multiple 171:178 arg1 inflammation 266:277 inflammation 266:277 inflammation 266:277 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 3 theme multiple 171:178 arg1 tumorigenesis 302:314 tumorigenesis 302:314 tumorigenesis 302:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 3 theme multiple 171:178 arg1 production 238:247 cytokine production 229:247 cytokine production 229:247 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 3 theme multiple 171:178 arg1 responses 287:295 immune responses 280:295 immune responses 280:295 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 4 theme immune 280:285 arg1 responses 287:295 immune responses 280:295 immune responses 280:295 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 7 5 theme novel 1355:1359 arg1 Y209 1366:1369 Y209 1366:1369 Y209 1366:1369 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 7 5 theme novel 1355:1359 arg1 one 1361:1363 a novel one 1353:1363 a novel one (Y209) 1353:1370 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 5 6 theme Fucα1-4 1046:1052 arg1 GlcNAc 1054:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 2 7 theme Post-translational 317:334 arg1 PTMs 351:354 PTMs 351:354 PTMs 351:354 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 2 7 theme Post-translational 317:334 arg1 modifications 336:348 Post-translational modifications 317:348 Post-translational modifications (PTMs) of OPN 317:362 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 4 8 theme OPN 780:782 arg1 regions 763:769 7 O-glycosylation regions 745:769 7 O-glycosylation regions of human OPN 745:782 We detected 28 O-glycopeptides from 7 O-glycosylation regions of human OPN, occupied by highly heterogeneous O-glycans. 25450502 0 9 theme osteopontin 120:130 arg1 sites 99:103 phosphorylation sites 83:103 phosphorylation sites of recombinant osteopontin 83:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 0 9 theme osteopontin 120:130 arg1 osteopontin 120:130 recombinant osteopontin 108:130 recombinant osteopontin 108:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 9 10 theme human 1624:1628 arg1 OPN 1630:1632 recombinant human OPN 1612:1632 recombinant human OPN 1612:1632 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 5 11 theme x/a 986:988 arg1 antigens 991:998 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 3 12 theme human 577:581 arg1 OPN 595:597 human recombinant OPN 577:597 human recombinant OPN 577:597 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 6 13 from information 1174:1184 arg1 sequence 1201:1208 the peptide sequence 1189:1208 the peptide sequence 1189:1208 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 3 14 gly O-glycopeptides 666:680 arg2 O-glycopeptides 666:680 O-glycopeptides 666:680 O-glycopeptides 666:680 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 7 15 theme mass 1311:1314 arg1 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 7 15 theme mass 1311:1314 arg1 spectrometry 1316:1327 reverse-phase liquid chromatography-tandem mass spectrometry 1268:1327 reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) 1268:1340 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 3 16 theme glycan 559:564 arg1 pattern 566:572 the overall glycan pattern 547:572 the overall glycan pattern of human recombinant OPN 547:597 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 5 17 theme T 902:902 arg1 Galβ1-3GalNAcα1- 914:929 Galβ1-3GalNAcα1- 914:929 Galβ1-3GalNAcα1- 914:929 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 17 theme T 902:902 arg1 antigens 904:911 T antigens 902:911 T antigens (Galβ1-3GalNAcα1-) 902:930 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 1 18 theme physiological 180:192 arg1 processes 211:219 multiple physiological and pathological processes 171:219 multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis 171:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 18 theme physiological 180:192 arg1 mineralization 250:263 mineralization 250:263 mineralization 250:263 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 18 theme physiological 180:192 arg1 inflammation 266:277 inflammation 266:277 inflammation 266:277 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 18 theme physiological 180:192 arg1 tumorigenesis 302:314 tumorigenesis 302:314 tumorigenesis 302:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 18 theme physiological 180:192 arg1 production 238:247 cytokine production 229:247 cytokine production 229:247 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 18 theme physiological 180:192 arg1 responses 287:295 immune responses 280:295 immune responses 280:295 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 10 19 theme Proteomics 1693:1702 arg1 part 1651:1654 part 1651:1654 part of a Special Issue entitled: Medical Proteomics 1651:1702 This article is part of a Special Issue entitled: Medical Proteomics. 25450502 9 20 theme further 1538:1544 arg1 research 1546:1553 further research 1538:1553 further research into the role of oligosaccharides and phosphorylation of recombinant human OPN 1538:1632 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 6 21 theme essential 1164:1172 arg1 information 1174:1184 essential information 1164:1184 essential information on the peptide sequence 1164:1208 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 10 22 theme entitled 1675:1682 arg1 Proteomics 1693:1702 a Special Issue entitled: Medical Proteomics 1659:1702 a Special Issue entitled: Medical Proteomics 1659:1702 This article is part of a Special Issue entitled: Medical Proteomics. 25450502 0 23 theme structural 14:23 arg1 characterization 25:40 Site-specific structural characterization 0:40 Site-specific structural characterization of O-glycosylation 0:59 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 7 24 theme phosphorylation 1227:1241 arg1 sites 1243:1247 26 phosphorylation sites 1224:1247 26 phosphorylation sites 1224:1247 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 4 25 theme human 774:778 arg1 OPN 780:782 human OPN 774:782 human OPN 774:782 We detected 28 O-glycopeptides from 7 O-glycosylation regions of human OPN, occupied by highly heterogeneous O-glycans. 25450502 7 26 theme chromatography-tandem 1289:1309 arg1 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 7 26 theme chromatography-tandem 1289:1309 arg1 spectrometry 1316:1327 reverse-phase liquid chromatography-tandem mass spectrometry 1268:1327 reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) 1268:1340 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 1 27 theme pathological 198:209 arg1 processes 211:219 multiple physiological and pathological processes 171:219 multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis 171:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 27 theme pathological 198:209 arg1 mineralization 250:263 mineralization 250:263 mineralization 250:263 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 27 theme pathological 198:209 arg1 inflammation 266:277 inflammation 266:277 inflammation 266:277 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 27 theme pathological 198:209 arg1 tumorigenesis 302:314 tumorigenesis 302:314 tumorigenesis 302:314 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 27 theme pathological 198:209 arg1 production 238:247 cytokine production 229:247 cytokine production 229:247 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 1 27 theme pathological 198:209 arg1 responses 287:295 immune responses 280:295 immune responses 280:295 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 0 28 theme Site-specific 0:12 arg1 characterization 25:40 Site-specific structural characterization 0:40 Site-specific structural characterization of O-glycosylation 0:59 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 9 29 theme recombinant 1612:1622 arg1 OPN 1630:1632 recombinant human OPN 1612:1632 recombinant human OPN 1612:1632 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 2 30 theme human 488:492 arg1 OPN 494:496 human OPN 488:496 human OPN 488:496 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 5 31 theme sialyl-T 953:960 arg1 antigens 962:969 sialyl-T antigens 953:969 sialyl-T antigens 953:969 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 3 32 theme recombinant 583:593 arg1 OPN 595:597 human recombinant OPN 577:597 human recombinant OPN 577:597 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 8 33 theme site-specific 1396:1408 arg1 characterization 1421:1436 a detailed, site-specific structural characterization 1384:1436 characterization 1421:1436 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 2 34 theme O-glycosylation 469:483 arg1 characterization 449:464 site-specific characterization 435:464 site-specific characterization of O-glycosylation in human OPN 435:496 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 4 35 theme heterogeneous 804:816 arg1 O-glycans 818:826 highly heterogeneous O-glycans 797:826 highly heterogeneous O-glycans 797:826 We detected 28 O-glycopeptides from 7 O-glycosylation regions of human OPN, occupied by highly heterogeneous O-glycans. 25450502 3 36 theme mass 685:688 arg1 MS 704:705 MS 704:705 MS 704:705 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 3 36 theme mass 685:688 arg1 spectrometry 690:701 mass spectrometry 685:701 mass spectrometry (MS) 685:706 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 3 37 theme OPN 595:597 arg1 pattern 566:572 the overall glycan pattern 547:572 the overall glycan pattern of human recombinant OPN 547:597 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 1 38 theme key 159:161 arg1 role 163:166 a key role 157:166 a key role 157:166 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 5 39 theme relevant 876:883 arg1 antigens 943:950 sialyl-Tn antigens 933:950 sialyl-Tn antigens 933:950 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 39 theme relevant 876:883 arg1 antigens 962:969 sialyl-T antigens 953:969 sialyl-T antigens 953:969 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 39 theme relevant 876:883 arg1 epitopes 885:892 functionally relevant epitopes 863:892 functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 863:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 39 theme relevant 876:883 arg1 antigens 991:998 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 39 theme relevant 876:883 arg1 antigens 904:911 T antigens 902:911 T antigens (Galβ1-3GalNAcα1-) 902:930 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 40 theme GlcNAc/Neuα2-3Galβ1-3 1024:1044 arg1 GlcNAc 1054:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 1 41 theme cytokine 229:236 arg1 production 238:247 cytokine production 229:247 cytokine production 229:247 Osteopontin (OPN) plays a key role in multiple physiological and pathological processes such as cytokine production, mineralization, inflammation, immune responses, and tumorigenesis. 25450502 0 42 theme O-glycosylation 45:59 arg1 characterization 25:40 Site-specific structural characterization 0:40 Site-specific structural characterization of O-glycosylation 0:59 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 0 42 theme O-glycosylation 45:59 arg1 identification 65:78 identification 65:78 identification of phosphorylation sites of recombinant osteopontin 65:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 3 43 theme O-glycopeptides 666:680 arg1 analysis 654:661 detailed structural analysis 634:661 detailed structural analysis of O-glycopeptides 634:680 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 2 44 theme site-specific 435:447 arg1 characterization 449:464 site-specific characterization 435:464 site-specific characterization of O-glycosylation in human OPN 435:496 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 5 45 contain carried 845:851 arg1 O-glycans 835:843 These O-glycans 829:843 These O-glycans 829:843 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg2 antigens 904:911 T antigens 902:911 T antigens (Galβ1-3GalNAcα1-) 902:930 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg3 part 857:860 part 857:860 part 857:860 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg2 antigens 943:950 sialyl-Tn antigens 933:950 sialyl-Tn antigens 933:950 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg2 antigens 962:969 sialyl-T antigens 953:969 sialyl-T antigens 953:969 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg2 antigens 991:998 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 45 contain carried 845:851 arg2 epitopes 885:892 functionally relevant epitopes 863:892 functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 863:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 3 46 theme lectin 607:612 arg1 array 614:618 a lectin array 605:618 a lectin array 605:618 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 6 47 theme O-glycopeptides 1094:1108 arg1 spectra 1069:1075 MS(3) spectra 1063:1075 MS(3) spectra of the generated O-glycopeptides 1063:1108 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 5 48 theme sialyl-Tn 933:941 arg1 antigens 943:950 sialyl-Tn antigens 933:950 sialyl-Tn antigens 933:950 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 8 49 theme structural 1410:1419 arg1 characterization 1421:1436 a detailed, site-specific structural characterization 1384:1436 characterization 1421:1436 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 5 50 dep antigens 991:998 arg1 Fucα1-3 1016:1022 Fucα1-3 1016:1022 Fucα1-3 1016:1022 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 5 50 dep antigens 991:998 arg1 [Neuα2-3Galβ1-4 1000:1014 [Neuα2-3Galβ1-4 1000:1014 sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc] 976:1060 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 6 51 theme peptide 1193:1199 arg1 sequence 1201:1208 the peptide sequence 1189:1208 the peptide sequence 1189:1208 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 8 52 theme OPN 1499:1501 arg1 sites 1490:1494 the phosphorylation sites 1470:1494 the phosphorylation sites of OPN 1470:1501 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 8 52 theme OPN 1499:1501 arg1 OPN 1499:1501 OPN 1499:1501 OPN 1499:1501 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 8 53 theme O-glycosylation 1441:1455 arg1 characterization 1421:1436 a detailed, site-specific structural characterization 1384:1436 characterization 1421:1436 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 7 54 theme reverse-phase 1268:1280 arg1 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 7 54 theme reverse-phase 1268:1280 arg1 spectrometry 1316:1327 reverse-phase liquid chromatography-tandem mass spectrometry 1268:1327 reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) 1268:1340 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 2 55 theme OPN 360:362 arg1 PTMs 351:354 PTMs 351:354 PTMs 351:354 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 2 55 theme OPN 360:362 arg1 modifications 336:348 Post-translational modifications 317:348 Post-translational modifications (PTMs) of OPN 317:362 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 6 56 theme backbone 1142:1149 arg1 cleavages 1117:1125 cleavages 1117:1125 cleavages of the peptide backbone 1117:1149 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 4 57 gly O-glycopeptides 724:738 arg2 O-glycopeptides 724:738 28 O-glycopeptides 721:738 28 O-glycopeptides 721:738 We detected 28 O-glycopeptides from 7 O-glycosylation regions of human OPN, occupied by highly heterogeneous O-glycans. 25450502 8 58 theme phosphorylation 1474:1488 arg1 sites 1490:1494 the phosphorylation sites 1470:1494 the phosphorylation sites of OPN 1470:1501 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 8 58 theme phosphorylation 1474:1488 arg1 OPN 1499:1501 OPN 1499:1501 OPN 1499:1501 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 6 59 theme MS 1063:1064 arg1 spectra 1069:1075 MS(3) spectra 1063:1075 MS(3) spectra of the generated O-glycopeptides 1063:1108 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 2 60 from characterization 449:464 arg1 OPN 494:496 human OPN 488:496 human OPN 488:496 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 4 61 theme O-glycosylation 747:761 arg1 regions 763:769 7 O-glycosylation regions 745:769 7 O-glycosylation regions of human OPN 745:782 We detected 28 O-glycopeptides from 7 O-glycosylation regions of human OPN, occupied by highly heterogeneous O-glycans. 25450502 7 62 theme liquid 1282:1287 arg1 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 RPLC-MS/MS 1330:1339 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 7 62 theme liquid 1282:1287 arg1 spectrometry 1316:1327 reverse-phase liquid chromatography-tandem mass spectrometry 1268:1327 reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) 1268:1340 Furthermore, 26 phosphorylation sites were identified by reverse-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), including a novel one (Y209). 25450502 0 63 theme sites 99:103 arg1 characterization 25:40 Site-specific structural characterization 0:40 Site-specific structural characterization of O-glycosylation 0:59 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 0 63 theme sites 99:103 arg1 identification 65:78 identification 65:78 identification of phosphorylation sites of recombinant osteopontin 65:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 6 64 theme generated 1084:1092 arg1 O-glycopeptides 1094:1108 the generated O-glycopeptides 1080:1108 the generated O-glycopeptides 1080:1108 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 3 65 theme detailed 634:641 arg1 analysis 654:661 detailed structural analysis 634:661 detailed structural analysis of O-glycopeptides 634:680 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 5 66 dep Fucα1-3 1016:1022 arg1 GlcNAc 1054:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc 1024:1059 These O-glycans carried, in part, functionally relevant epitopes such as T antigens (Galβ1-3GalNAcα1-), sialyl-Tn antigens, sialyl-T antigens, and sialyl-Le(x/a) antigens [Neuα2-3Galβ1-4(Fucα1-3)GlcNAc/Neuα2-3Galβ1-3(Fucα1-4)GlcNAc]. 25450502 2 67 theme biological 403:412 arg1 properties 414:423 biological properties 403:423 biological properties 403:423 Post-translational modifications (PTMs) of OPN significantly affect its structure and biological properties; however, site-specific characterization of O-glycosylation in human OPN has not been reported. 25450502 8 68 theme detailed 1386:1393 arg1 characterization 1421:1436 a detailed, site-specific structural characterization 1384:1436 characterization 1421:1436 We provide a detailed, site-specific structural characterization of O-glycosylation and identify the phosphorylation sites of OPN. 25450502 9 69 theme OPN 1630:1632 arg1 role 1564:1567 the role 1560:1567 the role of oligosaccharides 1560:1587 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 9 69 theme OPN 1630:1632 arg1 phosphorylation 1593:1607 phosphorylation 1593:1607 phosphorylation of recombinant human OPN 1593:1632 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 0 70 theme phosphorylation 83:97 arg1 sites 99:103 phosphorylation sites 83:103 phosphorylation sites of recombinant osteopontin 83:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 0 70 theme phosphorylation 83:97 arg1 osteopontin 120:130 recombinant osteopontin 108:130 recombinant osteopontin 108:130 Site-specific structural characterization of O-glycosylation and identification of phosphorylation sites of recombinant osteopontin. 25450502 6 71 theme peptide 1134:1140 arg1 backbone 1142:1149 the peptide backbone 1130:1149 the peptide backbone 1130:1149 MS(3) spectra of the generated O-glycopeptides showed cleavages of the peptide backbone and provided essential information on the peptide sequence. 25450502 3 72 theme structural 643:652 arg1 analysis 654:661 detailed structural analysis 634:661 detailed structural analysis of O-glycopeptides 634:680 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 9 73 theme oligosaccharides 1572:1587 arg1 role 1564:1567 the role 1560:1567 the role of oligosaccharides 1560:1587 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 9 73 theme oligosaccharides 1572:1587 arg1 phosphorylation 1593:1607 phosphorylation 1593:1607 phosphorylation of recombinant human OPN 1593:1632 These data lay the foundation for further research into the role of oligosaccharides and phosphorylation of recombinant human OPN. 25450502 3 74 theme overall 551:557 arg1 pattern 566:572 the overall glycan pattern 547:572 the overall glycan pattern of human recombinant OPN 547:597 In this work, we profiled the overall glycan pattern of human recombinant OPN using a lectin array and completed detailed structural analysis of O-glycopeptides by mass spectrometry (MS). 25450502 10 75 dep entitled 1675:1682 arg1 Medical 1685:1691 Medical 1685:1691 Medical 1685:1691 This article is part of a Special Issue entitled: Medical Proteomics. 25636227 8 0 gly O-glycosylation 1247:1261 arg2 sites 1263:1267 most previously known O-glycosylation sites 1225:1267 most previously known O-glycosylation sites 1225:1267 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 1 1 theme individual 189:198 arg1 glycoproteins 200:212 individual glycoproteins 189:212 individual glycoproteins 189:212 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 9 2 theme glycopeptides 1371:1383 arg1 total 1343:1347 a total 1341:1347 a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1341:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 0 3 theme sites 83:87 arg1 characterization 47:62 characterization 47:62 characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins 47:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 7 4 theme analytical 1139:1148 arg1 workflow 1150:1157 the analytical workflow 1135:1157 the analytical workflow 1135:1157 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 1 5 theme glycoproteins 200:212 arg1 challenge 225:233 a major challenge 217:233 a major challenge 217:233 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 1 5 theme glycoproteins 200:212 arg1 characterization 169:184 O-glycosylation-site characterization 148:184 O-glycosylation-site characterization of individual glycoproteins 148:212 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 4 6 theme O-glycan 624:631 arg1 chains 633:638 O-glycan chains 624:638 O-glycan chains 624:638 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 0 7 theme chorionic 98:106 arg1 gonadotropin 108:119 human chorionic gonadotropin 92:119 human chorionic gonadotropin 92:119 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 9 8 theme O-GalNAc-linked 1355:1369 arg1 glycopeptides 1371:1383 49 O-GalNAc-linked glycopeptides 1352:1383 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1352:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 4 9 used used 599:602 arg2 mixture 568:574 A mixture 566:574 A mixture of exoglycosidases 566:593 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 5 10 gly O-glycosylated 732:745 arg1 peptides 747:754 The O-glycosylated peptides 728:754 The O-glycosylated peptides 728:754 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 2 11 from difficult 358:366 arg1 studies 306:312 proteomic studies 296:312 proteomic studies 296:312 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 0 12 theme human 92:96 arg1 gonadotropin 108:119 human chorionic gonadotropin 92:119 human chorionic gonadotropin 92:119 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 0 13 from characterization 47:62 arg1 gonadotropin 108:119 human chorionic gonadotropin 92:119 human chorionic gonadotropin 92:119 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 0 13 from characterization 47:62 arg1 proteins 138:145 human-plasma proteins 125:145 human-plasma proteins 125:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 9 14 gly glycoproteins 1405:1417 arg1 glycoproteins 1405:1417 36 distinctive glycoproteins 1390:1417 36 distinctive glycoproteins in human plasma 1390:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 0 15 gly O-glycosylation 67:81 arg2 sites 83:87 O-glycosylation sites 67:87 O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins 67:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 3 16 theme proteins 526:533 arg1 proteome 556:563 the human-plasma proteome 539:563 the human-plasma proteome 539:563 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 3 16 theme proteins 526:533 arg1 sites 506:510 the O-glycosylation sites 486:510 the O-glycosylation sites of individual proteins 486:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 3 16 theme proteins 526:533 arg1 proteins 526:533 individual proteins 515:533 individual proteins 515:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 8 17 theme O-glycosylation 1247:1261 arg1 sites 1263:1267 most previously known O-glycosylation sites 1225:1267 most previously known O-glycosylation sites 1225:1267 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 6 18 theme LC-MS-MS 954:961 arg1 analysis 963:970 LC-MS-MS analysis 954:970 LC-MS-MS analysis 954:970 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 10 19 theme core 1550:1553 arg1 analysis 1538:1545 site analysis 1533:1545 site analysis of core 1 1533:1555 The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. 25636227 2 20 theme O-glycosylation-site 315:334 arg1 analysis 336:343 O-glycosylation-site analysis 315:343 O-glycosylation-site analysis 315:343 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 3 21 theme convenient 447:456 arg1 workflow 458:465 a rapid and convenient workflow 435:465 a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome 435:563 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 4 22 theme Ser 707:709 arg1 residues 718:725 the Ser or Thr residues 703:725 the Ser or Thr residues 703:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 5 23 theme liquid 790:795 arg1 LC-MS-MS 838:845 LC-MS-MS 838:845 LC-MS-MS 838:845 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 5 23 theme liquid 790:795 arg1 spectrometry 824:835 liquid chromatography-tandem mass spectrometry 790:835 liquid chromatography-tandem mass spectrometry (LC-MS-MS) 790:846 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 3 24 theme rapid 437:441 arg1 workflow 458:465 a rapid and convenient workflow 435:465 a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome 435:563 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 7 25 used used 1122:1125 arg2 gonadotropin 1098:1109 human chorionic gonadotropin 1082:1109 human chorionic gonadotropin (hCG) 1082:1115 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 7 25 used used 1122:1125 arg2 fetuin 1071:1076 Bovine fetuin 1064:1076 Bovine fetuin 1064:1076 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 7 25 used used 1122:1125 arg2 hCG 1112:1114 hCG 1112:1114 hCG 1112:1114 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 10 26 gly O-glycosylated 1572:1585 arg1 proteins 1587:1594 core 4 O-glycosylated proteins 1565:1594 core 4 O-glycosylated proteins 1565:1594 The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. 25636227 0 27 theme individual 5:14 arg1 proteins 16:23 individual proteins 5:23 individual proteins to proteomic samples 5:44 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 6 28 theme necessary 1019:1027 arg1 proteins 1054:1061 necessary for complex human-plasma proteins 1019:1061 necessary for complex human-plasma proteins 1019:1061 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 1 29 gly heterogeneity 250:262 arg1 structures 281:290 O-glycan core structures 267:290 O-glycan core structures 267:290 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 5 30 theme chromatography-tandem 797:817 arg1 LC-MS-MS 838:845 LC-MS-MS 838:845 LC-MS-MS 838:845 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 5 30 theme chromatography-tandem 797:817 arg1 spectrometry 824:835 liquid chromatography-tandem mass spectrometry 790:835 liquid chromatography-tandem mass spectrometry (LC-MS-MS) 790:846 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 10 31 theme O-glycosylated 1572:1585 arg1 proteins 1587:1594 core 4 O-glycosylated proteins 1565:1594 core 4 O-glycosylated proteins 1565:1594 The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. 25636227 1 32 theme major 219:223 arg1 challenge 225:233 a major challenge 217:233 a major challenge 217:233 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 1 32 theme major 219:223 arg1 characterization 169:184 O-glycosylation-site characterization 148:184 O-glycosylation-site characterization of individual glycoproteins 148:212 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 5 33 theme 203 Da 862:867 arg1 increase 874:881 the 203 Da mass increase 858:881 the 203 Da mass increase 858:881 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 6 34 theme human-plasma 1041:1052 arg1 proteins 1054:1061 necessary for complex human-plasma proteins 1019:1061 necessary for complex human-plasma proteins 1019:1061 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 10 35 theme site 1533:1536 arg1 analysis 1538:1545 site analysis 1533:1545 site analysis of core 1 1533:1555 The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. 25636227 5 36 theme mass 869:872 arg1 increase 874:881 the 203 Da mass increase 858:881 the 203 Da mass increase 858:881 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 8 37 theme known 1241:1245 arg1 sites 1263:1267 most previously known O-glycosylation sites 1225:1267 most previously known O-glycosylation sites 1225:1267 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 8 38 theme novel 1298:1302 arg1 sites 1304:1308 several novel sites 1290:1308 several novel sites 1290:1308 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 9 39 link O-GalNAc-linked 1355:1369 arg1 glycopeptides 1371:1383 49 O-GalNAc-linked glycopeptides 1352:1383 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1352:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 6 40 used used 896:899 arg2 Jacalin 884:890 Jacalin 884:890 Jacalin 884:890 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 9 41 theme human 1422:1426 arg1 plasma 1428:1433 human plasma 1422:1433 human plasma 1422:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 0 42 theme human-plasma 125:136 arg1 proteins 138:145 human-plasma proteins 125:145 human-plasma proteins 125:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 6 43 theme O-GalNAc 924:931 arg1 glycopeptides 933:945 O-GalNAc glycopeptides 924:945 O-GalNAc glycopeptides 924:945 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 2 44 from studies 306:312 arg1 difficult 358:366 difficult 358:366 difficult 358:366 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 3 45 theme human-plasma 543:554 arg1 proteome 556:563 the human-plasma proteome 539:563 the human-plasma proteome 539:563 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 4 46 theme Thr 714:716 arg1 residues 718:725 the Ser or Thr residues 703:725 the Ser or Thr residues 703:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 0 47 from sites 83:87 arg1 gonadotropin 108:119 human chorionic gonadotropin 92:119 human chorionic gonadotropin 92:119 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 0 47 from sites 83:87 arg1 proteins 138:145 human-plasma proteins 125:145 human-plasma proteins 125:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 0 48 dep characterization 47:62 arg1 proteins 16:23 individual proteins 5:23 individual proteins to proteomic samples 5:44 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 0 49 theme proteomic 28:36 arg1 samples 38:44 proteomic samples 28:44 proteomic samples 28:44 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 8 50 theme several 1290:1296 arg1 sites 1304:1308 several novel sites 1290:1308 several novel sites 1290:1308 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 0 51 from proteins 138:145 arg1 characterization 47:62 characterization 47:62 characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins 47:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 7 52 theme chorionic 1088:1096 arg1 hCG 1112:1114 hCG 1112:1114 hCG 1112:1114 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 7 52 theme chorionic 1088:1096 arg1 gonadotropin 1098:1109 human chorionic gonadotropin 1082:1109 human chorionic gonadotropin (hCG) 1082:1115 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 3 53 theme O-glycosylation 490:504 arg1 sites 506:510 the O-glycosylation sites 486:510 the O-glycosylation sites of individual proteins 486:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 3 53 theme O-glycosylation 490:504 arg1 proteins 526:533 individual proteins 515:533 individual proteins 515:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 9 54 from total 1343:1347 arg1 glycoproteins 1405:1417 36 distinctive glycoproteins 1390:1417 36 distinctive glycoproteins in human plasma 1390:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 3 55 gly O-glycosylation 490:504 arg2 sites 506:510 the O-glycosylation sites 486:510 the O-glycosylation sites of individual proteins 486:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 3 55 gly O-glycosylation 490:504 arg1 proteins 526:533 individual proteins 515:533 individual proteins 515:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 3 55 gly O-glycosylation 490:504 arg2 proteins 526:533 individual proteins 515:533 individual proteins 515:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 9 56 from glycoproteins 1405:1417 arg1 total 1343:1347 a total 1341:1347 a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1341:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 9 56 from glycoproteins 1405:1417 arg1 plasma 1428:1433 human plasma 1422:1433 human plasma 1422:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 9 56 from glycoproteins 1405:1417 arg1 glycopeptides 1371:1383 49 O-GalNAc-linked glycopeptides 1352:1383 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1352:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 6 57 theme individual 995:1004 arg1 proteins 1006:1013 individual proteins 995:1013 individual proteins 995:1013 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 9 58 theme blood 1334:1338 arg1 drop 1326:1329 only one drop 1317:1329 only one drop of blood 1317:1338 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 2 59 theme complexity 383:392 arg1 difficult 358:366 difficult 358:366 difficult 358:366 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 4 60 theme exoglycosidases 579:593 arg1 mixture 568:574 A mixture 566:574 A mixture of exoglycosidases 566:593 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 1 61 gly glycoproteins 200:212 arg1 glycoproteins 200:212 individual glycoproteins 189:212 individual glycoproteins 189:212 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 0 62 from gonadotropin 108:119 arg1 characterization 47:62 characterization 47:62 characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins 47:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 7 63 theme human 1082:1086 arg1 hCG 1112:1114 hCG 1112:1114 hCG 1112:1114 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 7 63 theme human 1082:1086 arg1 gonadotropin 1098:1109 human chorionic gonadotropin 1082:1109 human chorionic gonadotropin (hCG) 1082:1115 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 3 64 theme individual 515:524 arg1 proteins 526:533 individual proteins 515:533 individual proteins 515:533 In this work, we designed a rapid and convenient workflow for characterizing the O-glycosylation sites of individual proteins and the human-plasma proteome. 25636227 10 65 theme core 1565:1568 arg1 proteins 1587:1594 core 4 O-glycosylated proteins 1565:1594 core 4 O-glycosylated proteins 1565:1594 The approach described herein is simple, sensitive, and global for site analysis of core 1 through core 4 O-glycosylated proteins. 25636227 4 66 theme N-acetylgalacosamine 653:672 arg1 residue 683:689 an N-acetylgalacosamine (GalNAc) residue 650:689 an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues 650:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 5 67 theme O-glycosylated 732:745 arg1 peptides 747:754 The O-glycosylated peptides 728:754 The O-glycosylated peptides 728:754 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 4 68 attach attached 691:698 arg1 residues 718:725 the Ser or Thr residues 703:725 the Ser or Thr residues 703:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 4 68 attach attached 691:698 arg2 residue 683:689 an N-acetylgalacosamine (GalNAc) residue 650:689 an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues 650:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 2 69 theme proteomic 296:304 arg1 studies 306:312 proteomic studies 296:312 proteomic studies 296:312 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 7 70 theme Bovine 1064:1069 arg1 fetuin 1071:1076 Bovine fetuin 1064:1076 Bovine fetuin 1064:1076 Bovine fetuin and human chorionic gonadotropin (hCG) were used to test the analytical workflow. 25636227 1 71 theme O-glycosylation-site 148:167 arg1 challenge 225:233 a major challenge 217:233 a major challenge 217:233 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 1 71 theme O-glycosylation-site 148:167 arg1 characterization 169:184 O-glycosylation-site characterization 148:184 O-glycosylation-site characterization of individual glycoproteins 148:212 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 6 72 gly glycopeptides 933:945 arg2 glycopeptides 933:945 O-GalNAc glycopeptides 924:945 O-GalNAc glycopeptides 924:945 Jacalin was used to selectively isolate O-GalNAc glycopeptides before LC-MS-MS analysis, which is optional for individual proteins and necessary for complex human-plasma proteins. 25636227 1 73 theme O-glycan 267:274 arg1 structures 281:290 O-glycan core structures 267:290 O-glycan core structures 267:290 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 0 74 theme O-glycosylation 67:81 arg1 sites 83:87 O-glycosylation sites 67:87 O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins 67:145 From individual proteins to proteomic samples: characterization of O-glycosylation sites in human chorionic gonadotropin and human-plasma proteins. 25636227 9 75 theme distinctive 1393:1403 arg1 glycoproteins 1405:1417 36 distinctive glycoproteins 1390:1417 36 distinctive glycoproteins in human plasma 1390:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 4 76 theme GalNAc 675:680 arg1 residue 683:689 an N-acetylgalacosamine (GalNAc) residue 650:689 an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues 650:725 A mixture of exoglycosidases was used to partially remove O-glycan chains and leave an N-acetylgalacosamine (GalNAc) residue attached to the Ser or Thr residues. 25636227 2 77 theme sample 401:406 arg1 complexity 383:392 the complexity 379:392 the complexity of the sample 379:406 In proteomic studies, O-glycosylation-site analysis is even more difficult because of the complexity of the sample. 25636227 1 78 theme core 276:279 arg1 structures 281:290 O-glycan core structures 267:290 O-glycan core structures 267:290 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 25636227 5 79 theme mass 819:822 arg1 LC-MS-MS 838:845 LC-MS-MS 838:845 LC-MS-MS 838:845 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 5 79 theme mass 819:822 arg1 spectrometry 824:835 liquid chromatography-tandem mass spectrometry 790:835 liquid chromatography-tandem mass spectrometry (LC-MS-MS) 790:846 The O-glycosylated peptides could then be identified by using liquid chromatography-tandem mass spectrometry (LC-MS-MS) to detect the 203 Da mass increase. 25636227 9 80 gly glycopeptides 1371:1383 arg1 glycoproteins 1405:1417 36 distinctive glycoproteins 1390:1417 36 distinctive glycoproteins in human plasma 1390:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 9 80 gly glycopeptides 1371:1383 arg2 glycopeptides 1371:1383 49 O-GalNAc-linked glycopeptides 1352:1383 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma 1352:1433 Using only one drop of blood, a total of 49 O-GalNAc-linked glycopeptides from 36 distinctive glycoproteins in human plasma were identified unambiguously. 25636227 8 81 theme superior 1183:1190 arg1 sensitivity 1192:1202 superior sensitivity 1183:1202 superior sensitivity 1183:1202 The workflow indicated superior sensitivity by not only covering most previously known O-glycosylation sites but also discovering several novel sites. 25636227 1 82 theme structures 281:290 arg1 heterogeneity 250:262 the heterogeneity 246:262 the heterogeneity of O-glycan core structures 246:290 O-glycosylation-site characterization of individual glycoproteins is a major challenge because of the heterogeneity of O-glycan core structures. 27638310 0 0 theme rice 97:100 arg1 cells 102:106 suspension-cultured rice cells 77:106 suspension-cultured rice cells 77:106 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 10 1 theme plant 1568:1572 arg1 cells 1574:1578 plant cells 1568:1578 plant cells 1568:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 3 2 theme previous 496:503 arg1 reports 505:511 no previous reports 493:511 no previous reports of the O-glycosylation of rhGM-CSF from plant cells 493:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 0 3 theme suspension-cultured 77:95 arg1 cells 102:106 suspension-cultured rice cells 77:106 suspension-cultured rice cells 77:106 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 7 4 theme Pro 1145:1147 arg1 residues 1169:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro-to-hydroxyproline conversions occurred at Pro 2, Pro 6, and Pro 8 residues. 27638310 3 5 gly O-glycosylation 520:534 arg1 rhGM-CSF 539:546 rhGM-CSF 539:546 rhGM-CSF 539:546 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 3 6 from reports 505:511 arg1 cells 559:563 plant cells 553:563 plant cells 553:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 3 7 theme rhGM-CSF 539:546 arg1 O-glycosylation 520:534 the O-glycosylation 516:534 the O-glycosylation of rhGM-CSF from plant cells 516:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 2 8 link rice-cell-derived 279:295 arg1 rrhGM-CSF 307:315 rrhGM-CSF 307:315 rrhGM-CSF 307:315 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 8 link rice-cell-derived 279:295 arg1 rhGM-CSF 297:304 suspension-cultured rice-cell-derived rhGM-CSF 259:304 suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF) 259:316 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 6 9 theme glycopeptides 936:948 arg1 Analysis 907:914 Analysis 907:914 Analysis of pronase-digested glycopeptides 907:948 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 5 10 dep seven 830:834 arg1 to 827:828 to 827:828 to 827:828 Eight O-glycans comprising four arabinose residues with zero to seven galactose residues along with their relative quantities were analyzed. 27638310 7 11 theme Pro 1152:1154 arg1 residues 1169:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro-to-hydroxyproline conversions occurred at Pro 2, Pro 6, and Pro 8 residues. 27638310 3 12 from O-glycans 597:605 arg1 rrhGM-CSF 660:668 rrhGM-CSF 660:668 rrhGM-CSF 660:668 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 2 13 theme longer 331:336 arg1 time 354:357 a longer blood clearance time 329:357 a longer blood clearance time 329:357 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 6 14 theme pronase-digested 919:934 arg1 glycopeptides 936:948 pronase-digested glycopeptides 919:948 pronase-digested glycopeptides 919:948 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 3 15 theme O-glycosylation 520:534 arg1 reports 505:511 no previous reports 493:511 no previous reports of the O-glycosylation of rhGM-CSF from plant cells 493:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 9 16 theme biological 1474:1483 arg1 function 1485:1492 its biological function 1470:1492 its biological function 1470:1492 These findings indicate that O-glycans of rrhGM-CSF are essential for maintaining its structural stability and result in an extended in vivo half-life, but without affecting its biological function. 27638310 8 17 theme rrhGM-CSFs 1213:1222 arg1 preparation 1183:1193 The preparation 1179:1193 The preparation of deglycosylated rrhGM-CSFs 1179:1222 The preparation of deglycosylated rrhGM-CSFs revealed that deglycosylation greatly affects their α-helix structures. 27638310 9 18 theme rrhGM-CSF 1338:1346 arg1 O-glycans 1325:1333 O-glycans 1325:1333 O-glycans of rrhGM-CSF 1325:1346 These findings indicate that O-glycans of rrhGM-CSF are essential for maintaining its structural stability and result in an extended in vivo half-life, but without affecting its biological function. 27638310 5 19 theme arabinose 798:806 arg1 residues 808:815 four arabinose residues 793:815 four arabinose residues 793:815 Eight O-glycans comprising four arabinose residues with zero to seven galactose residues along with their relative quantities were analyzed. 27638310 2 20 theme rhGM-CSF 297:304 arg1 analysis 419:426 the analysis 415:426 the analysis of its N-glycans 415:443 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 20 theme rhGM-CSF 297:304 arg1 production 245:254 The production 241:254 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, 241:409 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 10 21 gly O-glycosylation 1527:1541 arg1 rhGM-CSF 1546:1553 rhGM-CSF 1546:1553 rhGM-CSF derived from plant cells 1546:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 3 22 gly O-glycosylation 608:622 arg2 sites 624:628 O-glycosylation sites 608:628 O-glycosylation sites 608:628 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 7 23 theme Pro 1163:1165 arg1 residues 1169:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro 2, Pro 6, and Pro 8 residues 1145:1176 Pro-to-hydroxyproline conversions occurred at Pro 2, Pro 6, and Pro 8 residues. 27638310 8 24 gly deglycosylated 1198:1211 arg1 rrhGM-CSFs 1213:1222 deglycosylated rrhGM-CSFs 1198:1222 deglycosylated rrhGM-CSFs 1198:1222 The preparation of deglycosylated rrhGM-CSFs revealed that deglycosylation greatly affects their α-helix structures. 27638310 3 25 from sites 624:628 arg1 rrhGM-CSF 660:668 rrhGM-CSF 660:668 rrhGM-CSF 660:668 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 2 26 theme yeast-derived 387:399 arg1 rhGM-CSF 401:408 yeast-derived rhGM-CSF 387:408 yeast-derived rhGM-CSF 387:408 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 4 27 theme O-glycans 720:728 arg1 presence 708:715 the presence 704:715 the presence of O-glycans comprising arabinose and galactose 704:763 Monosaccharide analysis revealed the presence of O-glycans comprising arabinose and galactose. 27638310 2 28 theme N-glycans 435:443 arg1 analysis 419:426 the analysis 415:426 the analysis of its N-glycans 415:443 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 28 theme N-glycans 435:443 arg1 production 245:254 The production 241:254 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, 241:409 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 10 29 from report 1513:1518 arg1 O-glycosylation 1527:1541 the O-glycosylation 1523:1541 the O-glycosylation of rhGM-CSF derived from plant cells 1523:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 3 30 theme plant 553:557 arg1 cells 559:563 plant cells 553:563 plant cells 553:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 7 31 theme Pro-to-hydroxyproline 1099:1119 arg1 conversions 1121:1131 Pro-to-hydroxyproline conversions 1099:1131 Pro-to-hydroxyproline conversions 1099:1131 Pro-to-hydroxyproline conversions occurred at Pro 2, Pro 6, and Pro 8 residues. 27638310 6 32 theme glycan 1050:1055 arg1 heterogeneity 1057:1069 glycan heterogeneity 1050:1069 glycan heterogeneity 1050:1069 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 0 33 theme O-glycosylation 14:28 arg1 sites 30:34 O-glycosylation sites 14:34 O-glycosylation sites 14:34 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 33 theme O-glycosylation 14:28 arg1 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 34 theme structural 119:128 arg1 role 130:133 their structural role 113:133 their structural role 113:133 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 2 35 link yeast-derived 387:399 arg1 rhGM-CSF 401:408 yeast-derived rhGM-CSF 387:408 yeast-derived rhGM-CSF 387:408 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 0 36 theme recombinant 39:49 arg1 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 10 37 attach derived 1555:1561 arg1 cells 1574:1578 plant cells 1568:1578 plant cells 1568:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 10 37 attach derived 1555:1561 arg2 rhGM-CSF 1546:1553 rhGM-CSF 1546:1553 rhGM-CSF derived from plant cells 1546:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 3 38 theme structural 641:650 arg1 role 652:655 their structural role 635:655 their structural role in rrhGM-CSF 635:668 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 6 39 attach attached 993:1000 arg2 O-glycans 969:977 the O-glycans 965:977 the O-glycans 965:977 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 6 39 attach attached 993:1000 arg1 Ser 1019:1021 Ser 9 1019:1023 Ser 9 1019:1023 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 6 39 attach attached 993:1000 arg1 Thr 1029:1031 Thr 10 1029:1034 Thr 10 1029:1034 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 6 39 attach attached 993:1000 arg1 Ser 1005:1007 Ser 5 1005:1009 Ser 5 1005:1009 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 6 39 attach attached 993:1000 arg1 Ser 1012:1014 Ser 7 1012:1016 Ser 7 1012:1016 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 9 40 theme structural 1382:1391 arg1 stability 1393:1401 its structural stability 1378:1401 its structural stability 1378:1401 These findings indicate that O-glycans of rrhGM-CSF are essential for maintaining its structural stability and result in an extended in vivo half-life, but without affecting its biological function. 27638310 10 41 theme first 1507:1511 arg1 This 1495:1498 This 1495:1498 This 1495:1498 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 10 41 theme first 1507:1511 arg1 report 1513:1518 the first report 1503:1518 the first report on the O-glycosylation of rhGM-CSF derived from plant cells 1503:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 5 42 theme galactose 836:844 arg1 residues 846:853 zero to seven galactose residues 822:853 zero to seven galactose residues along with their relative quantities 822:890 Eight O-glycans comprising four arabinose residues with zero to seven galactose residues along with their relative quantities were analyzed. 27638310 5 43 theme relative 872:879 arg1 quantities 881:890 their relative quantities 866:890 their relative quantities 866:890 Eight O-glycans comprising four arabinose residues with zero to seven galactose residues along with their relative quantities were analyzed. 27638310 0 44 theme GM-CSF 57:62 arg1 O-glycans 0:8 O-glycans 0:8 O-glycans 0:8 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 44 theme GM-CSF 57:62 arg1 sites 30:34 O-glycosylation sites 14:34 O-glycosylation sites 14:34 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 44 theme GM-CSF 57:62 arg1 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 8 45 theme deglycosylated 1198:1211 arg1 rrhGM-CSFs 1213:1222 deglycosylated rrhGM-CSFs 1198:1222 deglycosylated rrhGM-CSFs 1198:1222 The preparation of deglycosylated rrhGM-CSFs revealed that deglycosylation greatly affects their α-helix structures. 27638310 0 46 gly O-glycosylation 14:28 arg2 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 46 gly O-glycosylation 14:28 arg2 sites 30:34 O-glycosylation sites 14:34 O-glycosylation sites 14:34 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 46 gly O-glycosylation 14:28 arg1 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 0 47 theme human 51:55 arg1 GM-CSF 57:62 recombinant human GM-CSF 39:62 recombinant human GM-CSF 39:62 O-glycans and O-glycosylation sites of recombinant human GM-CSF derived from suspension-cultured rice cells, and their structural role. 27638310 2 48 contain has 325:327 arg2 bioactivity 372:382 the same bioactivity 363:382 the same bioactivity 363:382 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 48 contain has 325:327 arg1 rrhGM-CSF 307:315 rrhGM-CSF 307:315 rrhGM-CSF 307:315 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 48 contain has 325:327 arg1 rhGM-CSF 297:304 suspension-cultured rice-cell-derived rhGM-CSF 259:304 suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF) 259:316 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 48 contain has 325:327 arg2 time 354:357 a longer blood clearance time 329:357 a longer blood clearance time 329:357 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 10 49 theme rhGM-CSF 1546:1553 arg1 O-glycosylation 1527:1541 the O-glycosylation 1523:1541 the O-glycosylation of rhGM-CSF derived from plant cells 1523:1578 This is the first report on the O-glycosylation of rhGM-CSF derived from plant cells. 27638310 6 50 dep Ser 1005:1007 arg1 residues 1036:1043 residues 1036:1043 residues 1036:1043 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 2 51 theme clearance 344:352 arg1 time 354:357 a longer blood clearance time 329:357 a longer blood clearance time 329:357 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 3 52 theme O-glycosylation 608:622 arg1 sites 624:628 O-glycosylation sites 608:628 O-glycosylation sites 608:628 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 2 53 theme rice-cell-derived 279:295 arg1 rrhGM-CSF 307:315 rrhGM-CSF 307:315 rrhGM-CSF 307:315 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 53 theme rice-cell-derived 279:295 arg1 rhGM-CSF 297:304 suspension-cultured rice-cell-derived rhGM-CSF 259:304 suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF) 259:316 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 4 54 theme Monosaccharide 671:684 arg1 analysis 686:693 Monosaccharide analysis 671:693 Monosaccharide analysis 671:693 Monosaccharide analysis revealed the presence of O-glycans comprising arabinose and galactose. 27638310 2 55 theme blood 338:342 arg1 time 354:357 a longer blood clearance time 329:357 a longer blood clearance time 329:357 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 3 56 from cells 559:563 arg1 reports 505:511 no previous reports 493:511 no previous reports of the O-glycosylation of rhGM-CSF from plant cells 493:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 3 56 from cells 559:563 arg1 O-glycosylation 520:534 the O-glycosylation 516:534 the O-glycosylation of rhGM-CSF from plant cells 516:563 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 1 57 theme immunosuppressed 214:229 arg1 patients 231:238 immunosuppressed patients 214:238 immunosuppressed patients 214:238 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 2 58 theme suspension-cultured 259:277 arg1 rrhGM-CSF 307:315 rrhGM-CSF 307:315 rrhGM-CSF 307:315 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 2 58 theme suspension-cultured 259:277 arg1 rhGM-CSF 297:304 suspension-cultured rice-cell-derived rhGM-CSF 259:304 suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF) 259:316 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 1 59 from yeast 177:181 arg1 GM-CSF 154:159 Recombinant human GM-CSF 136:159 Recombinant human GM-CSF (rhGM-CSF) from yeast 136:181 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 1 59 from yeast 177:181 arg1 rhGM-CSF 162:169 rhGM-CSF 162:169 rhGM-CSF 162:169 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 1 60 theme Recombinant 136:146 arg1 GM-CSF 154:159 Recombinant human GM-CSF 136:159 Recombinant human GM-CSF (rhGM-CSF) from yeast 136:181 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 1 60 theme Recombinant 136:146 arg1 rhGM-CSF 162:169 rhGM-CSF 162:169 rhGM-CSF 162:169 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 3 61 from role 652:655 arg1 rrhGM-CSF 660:668 rrhGM-CSF 660:668 rrhGM-CSF 660:668 However, there are no previous reports of the O-glycosylation of rhGM-CSF from plant cells, and so this study investigated O-glycans, O-glycosylation sites, and their structural role in rrhGM-CSF. 27638310 8 62 theme α-helix 1276:1282 arg1 structures 1284:1293 their α-helix structures 1270:1293 their α-helix structures 1270:1293 The preparation of deglycosylated rrhGM-CSFs revealed that deglycosylation greatly affects their α-helix structures. 27638310 1 63 theme human 148:152 arg1 GM-CSF 154:159 Recombinant human GM-CSF 136:159 Recombinant human GM-CSF (rhGM-CSF) from yeast 136:181 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 1 63 theme human 148:152 arg1 rhGM-CSF 162:169 rhGM-CSF 162:169 rhGM-CSF 162:169 Recombinant human GM-CSF (rhGM-CSF) from yeast has been clinically applied to immunosuppressed patients. 27638310 6 64 gly glycopeptides 936:948 arg2 glycopeptides 936:948 pronase-digested glycopeptides 919:948 pronase-digested glycopeptides 919:948 Analysis of pronase-digested glycopeptides indicated that the O-glycans are partially attached to Ser 5, Ser 7, Ser 9, or Thr 10 residues, and glycan heterogeneity was confirmed at each site. 27638310 9 65 theme extended 1420:1427 arg1 half-life 1437:1445 an extended in vivo half-life 1417:1445 an extended in vivo half-life 1417:1445 These findings indicate that O-glycans of rrhGM-CSF are essential for maintaining its structural stability and result in an extended in vivo half-life, but without affecting its biological function. 27638310 2 66 theme same 367:370 arg1 bioactivity 372:382 the same bioactivity 363:382 the same bioactivity 363:382 The production of suspension-cultured rice-cell-derived rhGM-CSF (rrhGM-CSF), which has a longer blood clearance time and the same bioactivity as yeast-derived rhGM-CSF, and the analysis of its N-glycans have been reported recently. 27638310 9 67 theme in vivo 1429:1435 arg1 half-life 1437:1445 an extended in vivo half-life 1417:1445 an extended in vivo half-life 1417:1445 These findings indicate that O-glycans of rrhGM-CSF are essential for maintaining its structural stability and result in an extended in vivo half-life, but without affecting its biological function. 25094044 0 0 theme uridine 76:82 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 0 theme uridine 76:82 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 1 1 theme uridine 270:276 arg1 protein 261:267 a carrier protein 251:267 a carrier protein 251:267 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 1 theme uridine 270:276 arg1 5'-diphosphate-glucose 278:299 uridine 5'-diphosphate-glucose 270:299 uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 270:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 4 2 from site 660:663 arg1 UGGT1 734:738 UGGT1 734:738 UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens) 734:838 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 1 3 theme attachment 237:246 arg1 position 225:232 their position 219:232 their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 219:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 3 theme attachment 237:246 arg1 structures 204:213 glycan structures 197:213 glycan structures 197:213 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 4 dep 5'-diphosphate-glucose 278:299 arg1 UGGT1 336:340 UGGT1 336:340 UGGT1 336:340 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 4 dep 5'-diphosphate-glucose 278:299 arg1 glucosyltransferase 315:333 glycoprotein glucosyltransferase 302:333 uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 270:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 0 5 from structure 7:15 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 5 from structure 7:15 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 6 from bovine 163:168 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 2 7 gly glycosylated 440:451 arg1 polypeptides 453:464 newly synthesized glycosylated polypeptides 422:464 newly synthesized glycosylated polypeptides 422:464 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 3 8 attach attached 576:583 arg2 glycan 569:574 glycan 569:574 glycan attached to UGGT1 569:592 The structure of glycan attached to UGGT1, however, has not been investigated. 25094044 3 8 attach attached 576:583 arg1 UGGT1 588:592 UGGT1 588:592 UGGT1 588:592 The structure of glycan attached to UGGT1, however, has not been investigated. 25094044 4 9 theme Homo 826:829 arg1 sapiens 831:837 Homo sapiens 826:837 Homo sapiens 826:837 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 0 10 gly glycoprotein 62:73 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 10 gly glycoprotein 62:73 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 1 11 theme carrier 253:259 arg1 protein 261:267 a carrier protein 251:267 a carrier protein 251:267 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 11 theme carrier 253:259 arg1 5'-diphosphate-glucose 278:299 uridine 5'-diphosphate-glucose 270:299 uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 270:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 4 12 dep rat 754:756 arg1 norvegicus 766:775 Rattus norvegicus 759:775 Rattus norvegicus 759:775 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 0 13 theme glycoprotein 108:119 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 14 from glycosylation 29:41 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 14 from glycosylation 29:41 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 2 15 theme endoplasmic 473:483 arg1 reticulum 485:493 the endoplasmic reticulum 469:493 the endoplasmic reticulum 469:493 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 2 16 theme synthesized 428:438 arg1 polypeptides 453:464 newly synthesized glycosylated polypeptides 422:464 newly synthesized glycosylated polypeptides 422:464 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 2 17 theme glycosylated 440:451 arg1 polypeptides 453:464 newly synthesized glycosylated polypeptides 422:464 newly synthesized glycosylated polypeptides 422:464 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 1 18 theme mass 369:372 arg1 spectrometry 374:385 tandem mass spectrometry 362:385 tandem mass spectrometry 362:385 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 0 19 from porcine 154:160 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 20 theme Glycan 0:5 arg1 structure 7:15 Glycan structure 0:15 Glycan structure 0:15 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 3 21 theme glycan 569:574 arg1 structure 556:564 The structure 552:564 The structure of glycan attached to UGGT1 552:592 The structure of glycan attached to UGGT1, however, has not been investigated. 25094044 4 22 gly glycosylation 668:680 arg1 UGGT1 734:738 UGGT1 734:738 UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens) 734:838 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 22 gly glycosylation 668:680 arg2 structures 704:713 the glycan structures 693:713 the glycan structures (Hex5-8HexNAc2) 693:729 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 22 gly glycosylation 668:680 arg2 glycosylation 668:680 glycosylation 668:680 glycosylation (N269) 668:687 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 22 gly glycosylation 668:680 arg2 site 660:663 the site 656:663 the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens) 656:838 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 0 23 gly glycoprotein 108:119 arg1 glycoprotein 108:119 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 24 from human 175:179 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 4 25 dep Bos 802:804 arg1 taurus 806:811 Bos taurus 802:811 Bos taurus 802:811 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 26 theme glycan 697:702 arg1 Hex5-8HexNAc2 716:728 Hex5-8HexNAc2 716:728 Hex5-8HexNAc2 716:728 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 26 theme glycan 697:702 arg1 structures 704:713 the glycan structures 693:713 the glycan structures (Hex5-8HexNAc2) 693:729 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 2 27 theme folding 404:410 arg1 UGGT1 388:392 UGGT1 388:392 UGGT1 388:392 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 2 27 theme folding 404:410 arg1 sensor 412:417 a folding sensor 402:417 a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum 402:493 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 0 28 theme glycosylation 29:41 arg1 glycosylation 29:41 glycosylation 29:41 glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose 29:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 28 theme glycosylation 29:41 arg1 site 21:24 site 21:24 site 21:24 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 28 theme glycosylation 29:41 arg1 structure 7:15 Glycan structure 0:15 Glycan structure 0:15 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 1 29 theme glycan 197:202 arg1 structures 204:213 glycan structures 197:213 glycan structures 197:213 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 0 30 from glycoprotein 62:73 arg1 glycosylation 29:41 glycosylation 29:41 glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose 29:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 30 from glycoprotein 62:73 arg1 site 21:24 site 21:24 site 21:24 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 30 from glycoprotein 62:73 arg1 structure 7:15 Glycan structure 0:15 Glycan structure 0:15 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 1 31 theme glycoprotein 302:313 arg1 UGGT1 336:340 UGGT1 336:340 UGGT1 336:340 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 31 theme glycoprotein 302:313 arg1 glucosyltransferase 315:333 glycoprotein glucosyltransferase 302:333 uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 270:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 1 32 theme tandem 362:367 arg1 spectrometry 374:385 tandem mass spectrometry 362:385 tandem mass spectrometry 362:385 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 2 33 theme polypeptides 453:464 arg1 UGGT1 388:392 UGGT1 388:392 UGGT1 388:392 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 2 33 theme polypeptides 453:464 arg1 sensor 412:417 a folding sensor 402:417 a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum 402:493 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 4 34 theme glycosylation 668:680 arg1 site 660:663 the site 656:663 the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens) 656:838 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 34 theme glycosylation 668:680 arg1 structures 704:713 the glycan structures 693:713 the glycan structures (Hex5-8HexNAc2) 693:729 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 34 theme glycosylation 668:680 arg1 glycosylation 668:680 glycosylation 668:680 glycosylation (N269) 668:687 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 35 theme structures 704:713 arg1 site 660:663 the site 656:663 the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens) 656:838 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 35 theme structures 704:713 arg1 structures 704:713 the glycan structures 693:713 the glycan structures (Hex5-8HexNAc2) 693:729 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 4 35 theme structures 704:713 arg1 glycosylation 668:680 glycosylation 668:680 glycosylation (N269) 668:687 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25094044 0 36 gly glycosylation 29:41 arg2 glycosylation 29:41 glycosylation 29:41 glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose 29:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 36 gly glycosylation 29:41 arg2 site 21:24 site 21:24 site 21:24 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 36 gly glycosylation 29:41 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 36 gly glycosylation 29:41 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 1 37 gly glycoprotein 302:313 arg1 glycoprotein 302:313 glycoprotein glucosyltransferase 302:333 uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1) 270:341 Here we report glycan structures and their position of attachment to a carrier protein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferase (UGGT1), as detected using tandem mass spectrometry. 25094044 0 38 from rat 149:151 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 39 dep structure 7:15 arg1 glucosyltransferases 121:140 glycoprotein glucosyltransferases 1 108:142 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 0:180 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 2 40 gly glycosylated 538:549 arg1 transferase 504:514 the transferase itself 500:521 the transferase itself 500:521 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 0 41 theme ER-resident 50:60 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 41 theme ER-resident 50:60 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 42 from site 21:24 arg1 glycoprotein 62:73 the ER-resident glycoprotein 46:73 the ER-resident glycoprotein 46:73 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 0 42 from site 21:24 arg1 5'-diphosphate-glucose 84:105 uridine 5'-diphosphate-glucose 76:105 uridine 5'-diphosphate-glucose 76:105 Glycan structure and site of glycosylation in the ER-resident glycoprotein, uridine 5'-diphosphate-glucose: glycoprotein glucosyltransferases 1 from rat, porcine, bovine, and human. 25094044 2 43 from sensor 412:417 arg1 reticulum 485:493 the endoplasmic reticulum 469:493 the endoplasmic reticulum 469:493 UGGT1 acts as a folding sensor of newly synthesized glycosylated polypeptides in the endoplasmic reticulum, and the transferase itself is known to be glycosylated. 25094044 4 44 dep human 819:823 arg1 sapiens 831:837 Homo sapiens 826:837 Homo sapiens 826:837 In this study, we reveal the site of glycosylation (N269) and the glycan structures (Hex5-8HexNAc2) in UGGT1 obtained from rat (Rattus norvegicus), pig (Sus scrofa), cow (Bos taurus), and human (Homo sapiens). 25629924 2 0 gly glycosylated 241:252 arg1 protein 254:260 a highly glycosylated protein 232:260 a highly glycosylated protein 232:260 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 2 0 gly glycosylated 241:252 arg1 it 226:227 it 226:227 it 226:227 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 1 1 theme key 158:160 arg1 role 162:165 a key role 156:165 a key role 156:165 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 4 2 with structures 726:735 arg1 GlcNAc 751:756 terminal GlcNAc 742:756 terminal GlcNAc 742:756 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 5 3 from sites 959:963 arg1 compositions 938:949 16 compositions 935:949 16 compositions at five sites on the IgA heavy (H) chain 935:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 3 from sites 959:963 arg1 chain 986:990 the IgA heavy (H) chain 968:990 the IgA heavy (H) chain 968:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 3 from sites 959:963 arg1 compositions 817:828 nearly 30 glycan compositions 800:828 nearly 30 glycan compositions located at seven sites on the secretory component 800:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 3 from sites 959:963 arg1 compositions 885:896 six compositions 881:896 six compositions at a single site on the J chain 881:928 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 4 4 theme glycans 588:594 arg1 structures 618:627 biantennary structures 606:627 biantennary structures with one or more acidic Neu5Ac residues 606:667 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 4 4 theme glycans 588:594 arg1 majority 572:579 The majority 568:579 The majority of the glycans found 568:600 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 5 5 theme H 983:983 arg1 chain 986:990 the IgA heavy (H) chain 968:990 the IgA heavy (H) chain 968:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 7 6 theme O-linked 1171:1178 arg1 compositions 1187:1198 54 O-linked glycan compositions 1168:1198 54 O-linked glycan compositions located at the IgA1 hinge region (HR) 1168:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 1 7 theme immune 180:185 arg1 protection 187:196 immune protection 180:196 immune protection of the gut mucosa 180:214 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 5 8 theme glycan 810:815 arg1 compositions 817:828 nearly 30 glycan compositions 800:828 nearly 30 glycan compositions located at seven sites on the secretory component 800:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 0 9 from colostrum 69:77 arg1 A 56:56 secretory immunoglobulin A 31:56 secretory immunoglobulin A from human colostrum 31:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 0 9 from colostrum 69:77 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of secretory immunoglobulin A from human colostrum 0:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 7 10 theme O-glycopeptide 1290:1303 arg1 library 1305:1311 a theoretical O-glycopeptide library 1276:1311 a theoretical O-glycopeptide library 1276:1311 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 3 11 theme Glycopeptide 546:557 arg1 Finder 559:564 Glycopeptide Finder 546:564 Glycopeptide Finder 546:564 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 11 theme Glycopeptide 546:557 arg1 software 536:543 in-house software 527:543 in-house software (Glycopeptide Finder) 527:565 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 8 12 theme micro-heterogeneity 1403:1421 arg1 complexity 1382:1391 the complexity 1378:1391 the complexity of glycan micro-heterogeneity 1378:1421 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 6 13 theme nonspecific 1131:1141 arg1 proteases 1143:1151 nonspecific proteases 1131:1151 nonspecific proteases 1131:1151 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 5 14 theme heavy 976:980 arg1 chain 986:990 the IgA heavy (H) chain 968:990 the IgA heavy (H) chain 968:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 8 15 theme relative 1428:1435 arg1 quantitation 1437:1448 relative quantitation 1428:1448 relative quantitation of glycoforms for each glycosylation site on milk sIgA 1428:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 4 16 theme Neu5Ac 653:658 arg1 residues 660:667 one or more acidic Neu5Ac residues 634:667 one or more acidic Neu5Ac residues 634:667 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 5 17 theme secretory 860:868 arg1 component 870:878 the secretory component 856:878 the secretory component 856:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 4 18 theme truncated 708:716 arg1 structures 726:735 truncated complex structures 708:735 truncated complex structures with terminal GlcNAc 708:756 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 5 19 from sites 847:851 arg1 located 830:836 located 830:836 located 830:836 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 19 from sites 847:851 arg1 component 870:878 the secretory component 856:878 the secretory component 856:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 8 20 theme glycosylation 1473:1485 arg1 site 1487:1490 each glycosylation site 1468:1490 each glycosylation site on milk sIgA 1468:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 3 21 theme sIgA 425:428 arg1 glycosylation 408:420 the site-specific glycosylation 390:420 the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) 390:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 1 22 theme immunoglobulin 90:103 arg1 glycoprotein 125:136 a major glycoprotein 117:136 a major glycoprotein 117:136 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 22 theme immunoglobulin 90:103 arg1 sIgA 108:111 sIgA 108:111 sIgA 108:111 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 22 theme immunoglobulin 90:103 arg1 A 105:105 Secretory immunoglobulin A 80:105 Secretory immunoglobulin A (sIgA) 80:112 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 9 23 from glycosylation 1584:1596 arg1 complexes 1615:1623 large protein complexes 1601:1623 large protein complexes 1601:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 4 24 theme large 681:685 arg1 fraction 687:694 a large fraction 679:694 a large fraction 679:694 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 0 25 theme Site-specific 0:12 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of secretory immunoglobulin A from human colostrum 0:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 9 26 theme general 1539:1545 arg1 method 1547:1552 a general method 1537:1552 a general method for determining site-specific glycosylation in large protein complexes 1537:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 5 27 gly glycosites 768:777 arg2 glycosites 768:777 Multiple glycosites 759:777 Multiple glycosites 759:777 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 6 28 from extent 1075:1080 arg1 site 1104:1107 each site 1099:1107 each site 1099:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 0 29 theme immunoglobulin 41:54 arg1 A 56:56 secretory immunoglobulin A 31:56 secretory immunoglobulin A from human colostrum 31:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 0 30 gly glycosylation 14:26 arg1 A 56:56 secretory immunoglobulin A 31:56 secretory immunoglobulin A from human colostrum 31:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 8 31 theme comprehensive 1339:1351 arg1 report 1353:1358 the most comprehensive report 1330:1358 the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA 1330:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 6 32 from quantitation 1034:1045 arg1 site 1104:1107 each site 1099:1107 each site 1099:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 9 33 theme site-specific 1570:1582 arg1 glycosylation 1584:1596 site-specific glycosylation 1570:1596 site-specific glycosylation in large protein complexes 1570:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 7 34 theme IgA1 1215:1218 arg1 HR 1234:1235 HR 1234:1235 HR 1234:1235 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 7 34 theme IgA1 1215:1218 arg1 region 1226:1231 the IgA1 hinge region 1211:1231 the IgA1 hinge region (HR) 1211:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 4 35 theme terminal 742:749 arg1 GlcNAc 751:756 terminal GlcNAc 742:756 terminal GlcNAc 742:756 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 2 36 theme glycan 310:315 arg1 micro-heterogeneity 317:335 associated glycan micro-heterogeneity 299:335 associated glycan micro-heterogeneity 299:335 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 5 37 from site 910:913 arg1 compositions 938:949 16 compositions 935:949 16 compositions at five sites on the IgA heavy (H) chain 935:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 37 from site 910:913 arg1 chain 924:928 the J chain 918:928 the J chain 918:928 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 37 from site 910:913 arg1 compositions 817:828 nearly 30 glycan compositions 800:828 nearly 30 glycan compositions located at seven sites on the secretory component 800:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 37 from site 910:913 arg1 compositions 885:896 six compositions 881:896 six compositions at a single site on the J chain 881:928 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 38 theme located 830:836 arg1 compositions 817:828 nearly 30 glycan compositions 800:828 nearly 30 glycan compositions located at seven sites on the secretory component 800:878 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 1 39 theme mucosa 209:214 arg1 protection 187:196 immune protection 180:196 immune protection of the gut mucosa 180:214 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 8 40 from site 1487:1490 arg1 sIgA 1500:1503 milk sIgA 1495:1503 milk sIgA 1495:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 9 41 theme protein 1607:1613 arg1 complexes 1615:1623 large protein complexes 1601:1623 large protein complexes 1601:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 6 42 theme Site-specific 993:1005 arg1 heterogeneity 1007:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 9 43 gly glycosylation 1584:1596 arg1 complexes 1615:1623 large protein complexes 1601:1623 large protein complexes 1601:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 3 44 theme LC-MS 504:508 arg1 Finder 559:564 Glycopeptide Finder 546:564 Glycopeptide Finder 546:564 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 44 theme LC-MS 504:508 arg1 combination 489:499 a combination 487:499 a combination of LC-MS and LC-MS/MS 487:521 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 44 theme LC-MS 504:508 arg1 software 536:543 in-house software 527:543 in-house software (Glycopeptide Finder) 527:565 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 2 45 theme glycosylated 241:252 arg1 protein 254:260 a highly glycosylated protein 232:260 a highly glycosylated protein 232:260 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 2 45 theme glycosylated 241:252 arg1 it 226:227 it 226:227 it 226:227 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 3 46 attach isolated 430:437 arg1 n 461:461 n = 3 461:465 n = 3 461:465 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 46 attach isolated 430:437 arg1 colostrum 450:458 human colostrum 444:458 human colostrum (n = 3) 444:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 46 attach isolated 430:437 arg2 sIgA 425:428 sIgA 425:428 sIgA isolated from human colostrum (n = 3) 425:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 6 47 theme occupation 1085:1094 arg1 extent 1075:1080 the extent 1071:1080 the extent of occupation at each site 1071:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 47 theme occupation 1085:1094 arg1 heterogeneity 1007:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 47 theme occupation 1085:1094 arg1 quantitation 1034:1045 relative quantitation 1025:1045 relative quantitation 1025:1045 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 4 48 theme biantennary 606:616 arg1 structures 618:627 biantennary structures 606:627 biantennary structures with one or more acidic Neu5Ac residues 606:667 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 4 48 theme biantennary 606:616 arg1 majority 572:579 The majority 568:579 The majority of the glycans found 568:600 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 3 49 theme in-house 527:534 arg1 Finder 559:564 Glycopeptide Finder 546:564 Glycopeptide Finder 546:564 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 49 theme in-house 527:534 arg1 software 536:543 in-house software 527:543 in-house software (Glycopeptide Finder) 527:565 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 2 50 theme site-specific 267:279 arg1 glycosylation 281:293 its site-specific glycosylation 263:293 its site-specific glycosylation 263:293 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 7 51 link O-linked 1171:1178 arg1 compositions 1187:1198 54 O-linked glycan compositions 1168:1198 54 O-linked glycan compositions located at the IgA1 hinge region (HR) 1168:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 1 52 gly glycoprotein 125:136 arg1 glycoprotein 125:136 a major glycoprotein 117:136 a major glycoprotein 117:136 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 52 gly glycoprotein 125:136 arg1 A 105:105 Secretory immunoglobulin A 80:105 Secretory immunoglobulin A (sIgA) 80:112 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 8 53 theme glycan 1396:1401 arg1 micro-heterogeneity 1403:1421 glycan micro-heterogeneity 1396:1421 glycan micro-heterogeneity 1396:1421 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 7 54 gly O-glycopeptide 1290:1303 arg2 O-glycopeptide 1290:1303 a theoretical O-glycopeptide library 1276:1311 a theoretical O-glycopeptide library 1276:1311 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 5 55 theme IgA 972:974 arg1 chain 986:990 the IgA heavy (H) chain 968:990 the IgA heavy (H) chain 968:990 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 1 56 theme major 119:123 arg1 glycoprotein 125:136 a major glycoprotein 117:136 a major glycoprotein 117:136 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 56 theme major 119:123 arg1 A 105:105 Secretory immunoglobulin A 80:105 Secretory immunoglobulin A (sIgA) 80:112 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 3 57 theme LC-MS/MS 514:521 arg1 Finder 559:564 Glycopeptide Finder 546:564 Glycopeptide Finder 546:564 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 57 theme LC-MS/MS 514:521 arg1 combination 489:499 a combination 487:499 a combination of LC-MS and LC-MS/MS 487:521 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 57 theme LC-MS/MS 514:521 arg1 software 536:543 in-house software 527:543 in-house software (Glycopeptide Finder) 527:565 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 7 58 theme theoretical 1278:1288 arg1 library 1305:1311 a theoretical O-glycopeptide library 1276:1311 a theoretical O-glycopeptide library 1276:1311 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 8 59 gly glycosylation 1473:1485 arg2 site 1487:1490 each glycosylation site 1468:1490 each glycosylation site on milk sIgA 1468:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 8 60 theme glycoforms 1453:1462 arg1 quantitation 1437:1448 relative quantitation 1428:1448 relative quantitation of glycoforms for each glycosylation site on milk sIgA 1428:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 0 61 from glycosylation 14:26 arg1 colostrum 69:77 human colostrum 63:77 human colostrum 63:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 4 62 theme acidic 646:651 arg1 residues 660:667 one or more acidic Neu5Ac residues 634:667 one or more acidic Neu5Ac residues 634:667 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 6 63 gly heterogeneity 1007:1019 arg1 occupation 1085:1094 occupation 1085:1094 occupation at each site 1085:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 63 gly heterogeneity 1007:1019 arg1 composition 1055:1065 each composition 1050:1065 each composition 1050:1065 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 2 64 theme associated 299:308 arg1 micro-heterogeneity 317:335 associated glycan micro-heterogeneity 299:335 associated glycan micro-heterogeneity 299:335 Although it is a highly glycosylated protein, its site-specific glycosylation and associated glycan micro-heterogeneity have still not been fully elucidated. 25629924 1 65 theme Secretory 80:88 arg1 glycoprotein 125:136 a major glycoprotein 117:136 a major glycoprotein 117:136 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 65 theme Secretory 80:88 arg1 sIgA 108:111 sIgA 108:111 sIgA 108:111 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 1 65 theme Secretory 80:88 arg1 A 105:105 Secretory immunoglobulin A 80:105 Secretory immunoglobulin A (sIgA) 80:112 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 8 66 theme milk 1495:1498 arg1 sIgA 1500:1503 milk sIgA 1495:1503 milk sIgA 1495:1503 This represents the most comprehensive report to date detailing the complexity of glycan micro-heterogeneity with relative quantitation of glycoforms for each glycosylation site on milk sIgA. 25629924 4 67 theme complex 718:724 arg1 structures 726:735 truncated complex structures 708:735 truncated complex structures with terminal GlcNAc 708:756 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 6 68 from site 1104:1107 arg1 extent 1075:1080 the extent 1071:1080 the extent of occupation at each site 1071:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 68 from site 1104:1107 arg1 occupation 1085:1094 occupation 1085:1094 occupation at each site 1085:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 68 from site 1104:1107 arg1 heterogeneity 1007:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 68 from site 1104:1107 arg1 quantitation 1034:1045 relative quantitation 1025:1045 relative quantitation 1025:1045 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 5 69 theme Multiple 759:766 arg1 glycosites 768:777 Multiple glycosites 759:777 Multiple glycosites 759:777 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 1 70 from glycoprotein 125:136 arg1 milk 141:144 milk 141:144 milk 141:144 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 0 71 theme secretory 31:39 arg1 A 56:56 secretory immunoglobulin A 31:56 secretory immunoglobulin A from human colostrum 31:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 5 72 theme single 903:908 arg1 site 910:913 a single site 901:913 a single site on the J chain 901:928 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 5 73 theme J 922:922 arg1 chain 924:928 the J chain 918:928 the J chain 918:928 Multiple glycosites were identified with nearly 30 glycan compositions located at seven sites on the secretory component, six compositions at a single site on the J chain, and 16 compositions at five sites on the IgA heavy (H) chain. 25629924 6 74 theme relative 1025:1032 arg1 quantitation 1034:1045 relative quantitation 1025:1045 relative quantitation 1025:1045 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 0 75 theme A 56:56 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of secretory immunoglobulin A from human colostrum 0:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 7 76 theme hinge 1220:1224 arg1 HR 1234:1235 HR 1234:1235 HR 1234:1235 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 7 76 theme hinge 1220:1224 arg1 region 1226:1231 the IgA1 hinge region 1211:1231 the IgA1 hinge region (HR) 1211:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 3 77 theme site-specific 394:406 arg1 glycosylation 408:420 the site-specific glycosylation 390:420 the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) 390:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 4 78 with structures 618:627 arg1 residues 660:667 one or more acidic Neu5Ac residues 634:667 one or more acidic Neu5Ac residues 634:667 The majority of the glycans found are biantennary structures with one or more acidic Neu5Ac residues; however, a large fraction belonged to truncated complex structures with terminal GlcNAc. 25629924 3 79 theme human 444:448 arg1 n 461:461 n = 3 461:465 n = 3 461:465 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 79 theme human 444:448 arg1 colostrum 450:458 human colostrum 444:458 human colostrum (n = 3) 444:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 0 80 theme human 63:67 arg1 colostrum 69:77 human colostrum 63:77 human colostrum 63:77 Site-specific glycosylation of secretory immunoglobulin A from human colostrum. 25629924 7 81 from region 1226:1231 arg1 located 1200:1206 located 1200:1206 located 1200:1206 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 6 82 theme composition 1055:1065 arg1 extent 1075:1080 the extent 1071:1080 the extent of occupation at each site 1071:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 82 theme composition 1055:1065 arg1 heterogeneity 1007:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity 993:1019 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 6 82 theme composition 1055:1065 arg1 quantitation 1034:1045 relative quantitation 1025:1045 relative quantitation 1025:1045 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 7 83 theme located 1200:1206 arg1 compositions 1187:1198 54 O-linked glycan compositions 1168:1198 54 O-linked glycan compositions located at the IgA1 hinge region (HR) 1168:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 6 84 from heterogeneity 1007:1019 arg1 site 1104:1107 each site 1099:1107 each site 1099:1107 Site-specific heterogeneity and relative quantitation of each composition and the extent of occupation at each site were determined using nonspecific proteases. 25629924 3 85 theme = 463:463 arg1 n 461:461 n = 3 461:465 n = 3 461:465 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 85 theme = 463:463 arg1 colostrum 450:458 human colostrum 444:458 human colostrum (n = 3) 444:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 1 86 theme gut 205:207 arg1 mucosa 209:214 the gut mucosa 201:214 the gut mucosa 201:214 Secretory immunoglobulin A (sIgA) is a major glycoprotein in milk and plays a key role in mediating immune protection of the gut mucosa. 25629924 9 87 theme large 1601:1605 arg1 complexes 1615:1623 large protein complexes 1601:1623 large protein complexes 1601:1623 This strategy further provides a general method for determining site-specific glycosylation in large protein complexes. 25629924 7 88 theme glycan 1180:1185 arg1 compositions 1187:1198 54 O-linked glycan compositions 1168:1198 54 O-linked glycan compositions located at the IgA1 hinge region (HR) 1168:1236 Additionally, 54 O-linked glycan compositions located at the IgA1 hinge region (HR) were identified by comparison against a theoretical O-glycopeptide library. 25629924 3 89 gly glycosylation 408:420 arg1 sIgA 425:428 sIgA 425:428 sIgA isolated from human colostrum (n = 3) 425:466 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 25629924 3 89 gly glycosylation 408:420 arg1 study 383:387 this study 378:387 this study 378:387 In this study, the site-specific glycosylation of sIgA isolated from human colostrum (n = 3) was analyzed using a combination of LC-MS and LC-MS/MS and in-house software (Glycopeptide Finder). 26701617 10 0 from pastoris 1798:1805 arg1 production 1704:1713 production 1704:1713 production 1704:1713 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 0 from pastoris 1798:1805 arg1 design 1693:1698 design 1693:1698 design 1693:1698 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 1 theme expressed 1732:1740 arg1 PDGF-BB 1784:1790 homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB 1718:1790 homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications 1718:1837 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 7 2 theme mitogenic 1262:1270 arg1 activity 1272:1279 rhPDGF-BB mitogenic activity 1252:1279 rhPDGF-BB mitogenic activity 1252:1279 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 6 3 theme site-directed 920:932 arg1 mutagenesis 934:944 site-directed mutagenesis 920:944 site-directed mutagenesis 920:944 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 3 4 theme O-linked 612:619 arg1 glycans 621:627 O-linked glycans 612:627 O-linked glycans 612:627 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 8 5 theme PDGF-BBΔGly 1314:1324 arg1 mutant 1326:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. 26701617 8 6 theme wild-type 1388:1396 arg1 form 1398:1401 the wild-type form 1384:1401 the wild-type form 1384:1401 In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. 26701617 10 7 theme PDGF-BB 1612:1618 arg1 structure 1620:1628 PDGF-BB structure 1612:1628 PDGF-BB structure 1612:1628 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 1 8 theme heterologous 189:200 arg1 expression 210:219 heterologous protein expression 189:219 heterologous protein expression 189:219 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 6 9 theme first 994:998 arg1 time 1000:1003 the first time 990:1003 the first time 990:1003 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 10 10 dep expressed 1732:1740 arg1 human-type 1760:1769 human-type 1760:1769 human-type 1760:1769 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 10 dep expressed 1732:1740 arg1 unglycosylated 1742:1755 unglycosylated 1742:1755 unglycosylated 1742:1755 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 10 dep expressed 1732:1740 arg1 glycosylated 1771:1782 glycosylated 1771:1782 glycosylated 1771:1782 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 7 11 gly glycosylation 1217:1229 arg2 sites 1231:1235 glycosylation sites 1217:1235 glycosylation sites 1217:1235 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 10 12 theme pharmaceutical 1811:1824 arg1 applications 1826:1837 pharmaceutical applications 1811:1837 pharmaceutical applications 1811:1837 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 7 13 theme glycosylation 1217:1229 arg1 sites 1231:1235 glycosylation sites 1217:1235 glycosylation sites 1217:1235 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 10 14 theme glycosylation 1595:1607 arg1 effect 1585:1590 the effect 1581:1590 the effect of glycosylation on PDGF-BB structure and biological activity 1581:1652 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 2 15 theme unexpected 368:377 arg1 alterations 379:389 unexpected alterations 368:389 unexpected alterations in protein structure and function 368:423 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 6 16 theme major 1065:1069 arg1 residues 1031:1038 two threonine residues 1017:1038 two threonine residues at the C-terminus 1017:1056 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 6 16 theme major 1065:1069 arg1 sites 1087:1091 the major O-glycosylation sites 1061:1091 the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris 1061:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 5 17 theme secretion 806:814 arg1 process 816:822 the secretion process 802:822 the secretion process 802:822 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 7 18 theme protein 1182:1188 arg1 expression 1190:1199 heterogeneous protein expression 1168:1199 heterogeneous protein expression 1168:1199 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 1 19 theme Pichia 145:150 arg1 pastoris 152:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 1 19 theme Pichia 145:150 arg1 system 178:183 a widely used system 164:183 a widely used system for heterologous protein expression 164:219 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 0 20 from pastoris 129:136 arg1 Glycosylation 50:62 Glycosylation 50:62 Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 50:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 9 21 gly glycosylated 1463:1474 arg1 PDGF-BB 1476:1482 glycosylated PDGF-BB 1463:1482 glycosylated PDGF-BB 1463:1482 Furthermore, antiserum against PDGF-BBΔGly also recognized glycosylated PDGF-BB, indicating that protein immunogenicity was unaltered by glycosylation. 26701617 0 22 theme Human 79:83 arg1 Factor-BB 109:117 Recombinant Human Platelet-Derived Growth Factor-BB 67:117 Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 67:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 3 23 from effects 601:607 arg1 activity 651:658 PDGF-BB functional activity 632:658 PDGF-BB functional activity 632:658 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 0 24 theme Growth 102:107 arg1 Factor-BB 109:117 Recombinant Human Platelet-Derived Growth Factor-BB 67:117 Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 67:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 3 25 gly glycosylation 464:476 arg2 sites 478:482 glycosylation sites 464:482 glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 464:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 2 26 from alterations 379:389 arg1 structure 402:410 protein structure 394:410 protein structure 394:410 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 2 26 from alterations 379:389 arg1 function 416:423 function 416:423 function 416:423 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 2 27 theme recombinant 336:346 arg1 proteins 348:355 recombinant proteins 336:355 recombinant proteins 336:355 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 3 28 theme human 499:503 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 28 theme human 499:503 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 5 29 theme putative 837:844 arg1 sites 862:866 putative O-glycosylation sites 837:866 putative O-glycosylation sites 837:866 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 3 30 theme study 442:446 arg1 aim 430:432 The aim 426:432 The aim of this study 426:446 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 0 31 theme Platelet-Derived 85:100 arg1 Factor-BB 109:117 Recombinant Human Platelet-Derived Growth Factor-BB 67:117 Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 67:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 9 32 theme protein 1501:1507 arg1 immunogenicity 1509:1522 protein immunogenicity 1501:1522 protein immunogenicity 1501:1522 Furthermore, antiserum against PDGF-BBΔGly also recognized glycosylated PDGF-BB, indicating that protein immunogenicity was unaltered by glycosylation. 26701617 0 33 theme Functional 19:28 arg1 Characterization 30:45 Functional Characterization 19:45 Functional Characterization 19:45 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 3 34 theme glycosylation 464:476 arg1 sites 478:482 glycosylation sites 464:482 glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 464:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 4 35 contain contains 719:726 arg1 PDGF-BB 661:667 PDGF-BB 661:667 PDGF-BB secreted by P. pastoris 661:691 PDGF-BB secreted by P. pastoris is very heterogeneous and contains multiple isoforms. 26701617 4 35 contain contains 719:726 arg2 isoforms 737:744 multiple isoforms 728:744 multiple isoforms 728:744 PDGF-BB secreted by P. pastoris is very heterogeneous and contains multiple isoforms. 26701617 3 36 theme growth 522:527 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 36 theme growth 522:527 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 6 37 from C-terminus 1047:1056 arg1 sites 1087:1091 the major O-glycosylation sites 1061:1091 the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris 1061:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 6 37 from C-terminus 1047:1056 arg1 residues 1031:1038 two threonine residues 1017:1038 two threonine residues at the C-terminus 1017:1056 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 10 38 from PDGF-BB 1784:1790 arg1 pastoris 1798:1805 P. pastoris 1795:1805 P. pastoris 1795:1805 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 2 39 theme protein 394:400 arg1 structure 402:410 protein structure 394:410 protein structure 394:410 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 6 40 theme LC/MS 966:970 arg1 analysis 972:979 high-resolution LC/MS analysis 950:979 high-resolution LC/MS analysis 950:979 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 0 41 gly Glycosylation 50:62 arg1 Factor-BB 109:117 Recombinant Human Platelet-Derived Growth Factor-BB 67:117 Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 67:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 7 42 theme rhPDGF-BB 1252:1260 arg1 activity 1272:1279 rhPDGF-BB mitogenic activity 1252:1279 rhPDGF-BB mitogenic activity 1252:1279 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 6 43 theme threonine 1021:1029 arg1 sites 1087:1091 the major O-glycosylation sites 1061:1091 the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris 1061:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 6 43 theme threonine 1021:1029 arg1 residues 1031:1038 two threonine residues 1017:1038 two threonine residues at the C-terminus 1017:1056 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 2 44 theme proteins 348:355 arg1 application 321:331 pharmaceutical application 306:331 pharmaceutical application of recombinant proteins 306:355 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 10 45 theme PDGF-BB 1784:1790 arg1 production 1704:1713 production 1704:1713 production 1704:1713 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 45 theme PDGF-BB 1784:1790 arg1 design 1693:1698 design 1693:1698 design 1693:1698 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 7 46 theme sites 1231:1235 arg1 removal 1206:1212 the removal 1202:1212 the removal of glycosylation sites 1202:1235 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 3 47 theme possible 592:599 arg1 effects 601:607 possible effects 592:607 possible effects of O-linked glycans on PDGF-BB functional activity 592:658 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 8 48 theme unglycosylated 1299:1312 arg1 mutant 1326:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. 26701617 10 49 from production 1704:1713 arg1 pastoris 1798:1805 P. pastoris 1795:1805 P. pastoris 1795:1805 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 4 50 theme multiple 728:735 arg1 isoforms 737:744 multiple isoforms 728:744 multiple isoforms 728:744 PDGF-BB secreted by P. pastoris is very heterogeneous and contains multiple isoforms. 26701617 5 51 theme glycosylation 874:886 arg1 staining 888:895 glycosylation staining 874:895 glycosylation staining 874:895 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 3 52 theme PDGF-BB 632:638 arg1 activity 651:658 PDGF-BB functional activity 632:658 PDGF-BB functional activity 632:658 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 1 53 theme protein 202:208 arg1 expression 210:219 heterologous protein expression 189:219 heterologous protein expression 189:219 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 3 54 link platelet-derived 505:520 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 54 link platelet-derived 505:520 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 6 55 gly O-glycosylation 1071:1085 arg2 sites 1087:1091 the major O-glycosylation sites 1061:1091 the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris 1061:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 6 55 gly O-glycosylation 1071:1085 arg2 residues 1031:1038 two threonine residues 1017:1038 two threonine residues at the C-terminus 1017:1056 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 3 56 theme glycans 621:627 arg1 effects 601:607 possible effects 592:607 possible effects of O-linked glycans on PDGF-BB functional activity 592:658 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 57 theme functional 640:649 arg1 activity 651:658 PDGF-BB functional activity 632:658 PDGF-BB functional activity 632:658 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 6 58 theme O-glycosylation 1071:1085 arg1 residues 1031:1038 two threonine residues 1017:1038 two threonine residues at the C-terminus 1017:1056 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 6 58 theme O-glycosylation 1071:1085 arg1 sites 1087:1091 the major O-glycosylation sites 1061:1091 the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris 1061:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 0 59 from Factor-BB 109:117 arg1 pastoris 129:136 Pichia pastoris 122:136 Pichia pastoris 122:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 8 60 theme comparable 1362:1371 arg1 immunogenicity 1347:1360 the immunogenicity 1343:1360 the immunogenicity comparable to that of the wild-type form 1343:1401 In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. 26701617 7 61 theme heterogeneous 1168:1180 arg1 expression 1190:1199 heterogeneous protein expression 1168:1199 heterogeneous protein expression 1168:1199 Although O-glycosylation resulted in heterogeneous protein expression, the removal of glycosylation sites did not affect rhPDGF-BB mitogenic activity. 26701617 1 62 theme Yeast 139:143 arg1 pastoris 152:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 1 62 theme Yeast 139:143 arg1 system 178:183 a widely used system 164:183 a widely used system for heterologous protein expression 164:219 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 9 63 theme glycosylated 1463:1474 arg1 PDGF-BB 1476:1482 glycosylated PDGF-BB 1463:1482 glycosylated PDGF-BB 1463:1482 Furthermore, antiserum against PDGF-BBΔGly also recognized glycosylated PDGF-BB, indicating that protein immunogenicity was unaltered by glycosylation. 26701617 5 64 gly O-glycosylated 780:793 arg1 PDGF-BB 768:774 PDGF-BB 768:774 PDGF-BB 768:774 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 2 65 theme pharmaceutical 306:319 arg1 application 321:331 pharmaceutical application 306:331 pharmaceutical application of recombinant proteins 306:355 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 5 66 theme O-glycosylation 846:860 arg1 sites 862:866 putative O-glycosylation sites 837:866 putative O-glycosylation sites 837:866 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 10 67 from design 1693:1698 arg1 pastoris 1798:1805 P. pastoris 1795:1805 P. pastoris 1795:1805 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 0 68 theme Factor-BB 109:117 arg1 Glycosylation 50:62 Glycosylation 50:62 Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 50:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 1 69 theme used 173:176 arg1 pastoris 152:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris 139:159 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 1 69 theme used 173:176 arg1 system 178:183 a widely used system 164:183 a widely used system for heterologous protein expression 164:219 Yeast Pichia pastoris is a widely used system for heterologous protein expression. 26701617 6 70 theme P. 1118:1119 arg1 pastoris 1121:1128 P. pastoris 1118:1128 P. pastoris 1118:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 10 71 theme biological 1634:1643 arg1 activity 1645:1652 biological activity 1634:1652 biological activity 1634:1652 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 0 72 theme Pichia 122:127 arg1 pastoris 129:136 Pichia pastoris 122:136 Pichia pastoris 122:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 3 73 from sites 478:482 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 73 from sites 478:482 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 5 74 gly O-glycosylation 846:860 arg2 sites 862:866 putative O-glycosylation sites 837:866 putative O-glycosylation sites 837:866 We demonstrated that PDGF-BB was O-glycosylated during the secretion process and detected putative O-glycosylation sites using glycosylation staining and immunoblotting. 26701617 10 75 dep design 1693:1698 arg1 the 1689:1691 the 1689:1691 the 1689:1691 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 3 76 link O-linked 612:619 arg1 glycans 621:627 O-linked glycans 612:627 O-linked glycans 612:627 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 0 77 from Glycosylation 50:62 arg1 pastoris 129:136 Pichia pastoris 122:136 Pichia pastoris 122:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 3 78 theme platelet-derived 505:520 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 78 theme platelet-derived 505:520 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 79 theme recombinant 487:497 arg1 factor-BB 529:537 recombinant human platelet-derived growth factor-BB 487:537 recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris 487:573 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 3 79 theme recombinant 487:497 arg1 rhPDGF-BB 540:548 rhPDGF-BB 540:548 rhPDGF-BB 540:548 The aim of this study was to identify glycosylation sites on recombinant human platelet-derived growth factor-BB (rhPDGF-BB) secreted by P. pastoris, and investigate possible effects of O-linked glycans on PDGF-BB functional activity. 26701617 10 80 gly glycosylation 1595:1607 arg1 PDGF-BB 1612:1618 PDGF-BB structure 1612:1628 PDGF-BB structure 1612:1628 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 0 81 theme Glycosylation 50:62 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 0 81 theme Glycosylation 50:62 arg1 Characterization 30:45 Functional Characterization 19:45 Functional Characterization 19:45 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 6 82 from sites 1087:1091 arg1 rhPDGF-BB 1096:1104 rhPDGF-BB 1096:1104 rhPDGF-BB produced in P. pastoris 1096:1128 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 0 83 theme Recombinant 67:77 arg1 Factor-BB 109:117 Recombinant Human Platelet-Derived Growth Factor-BB 67:117 Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris 67:136 Identification and Functional Characterization of Glycosylation of Recombinant Human Platelet-Derived Growth Factor-BB in Pichia pastoris. 26701617 2 84 theme post-translational 231:248 arg1 modifications 250:262 post-translational modifications 231:262 post-translational modifications 231:262 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 2 84 theme post-translational 231:248 arg1 glycosylation 276:288 glycosylation 276:288 especially glycosylation 265:288 However, post-translational modifications, especially glycosylation, usually impede pharmaceutical application of recombinant proteins because of unexpected alterations in protein structure and function. 26701617 10 85 from effect 1585:1590 arg1 structure 1620:1628 PDGF-BB structure 1612:1628 PDGF-BB structure 1612:1628 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 10 85 from effect 1585:1590 arg1 activity 1645:1652 biological activity 1634:1652 biological activity 1634:1652 These findings elucidate the effect of glycosylation on PDGF-BB structure and biological activity, and can potentially contribute to the design and production of homogeneously expressed unglycosylated or human-type glycosylated PDGF-BB in P. pastoris for pharmaceutical applications. 26701617 4 86 theme P. 681:682 arg1 pastoris 684:691 P. pastoris 681:691 P. pastoris 681:691 PDGF-BB secreted by P. pastoris is very heterogeneous and contains multiple isoforms. 26701617 6 87 theme high-resolution 950:964 arg1 analysis 972:979 high-resolution LC/MS analysis 950:979 high-resolution LC/MS analysis 950:979 By site-directed mutagenesis and high-resolution LC/MS analysis, we, for the first time, identified two threonine residues at the C-terminus as the major O-glycosylation sites on rhPDGF-BB produced in P. pastoris. 26701617 8 88 gly unglycosylated 1299:1312 arg1 mutant 1326:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 the unglycosylated PDGF-BBΔGly mutant 1295:1331 In addition, the unglycosylated PDGF-BBΔGly mutant exhibited the immunogenicity comparable to that of the wild-type form. 26924641 3 0 theme mouse 743:747 arg1 plasma 749:754 mouse plasma 743:754 mouse plasma 743:754 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 6 1 theme sialylation 1494:1504 arg1 " 1505:1505 "branching sialylation" 1483:1505 "branching sialylation" 1483:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 3 2 from effect 683:688 arg1 N-glycosylation 724:738 N-glycosylation 724:738 N-glycosylation in mouse plasma and peritoneal fluid 724:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 3 theme spectrometry 575:586 arg1 profiling 623:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 1 4 theme inflammation 281:292 arg1 initiation 236:245 initiation 236:245 initiation 236:245 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 4 5 theme type 921:924 arg1 N-glycans 926:934 diantennary and triantennary complex type N-glycans 884:934 diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 884:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 3 6 theme time-of-flight 555:568 arg1 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 6 theme time-of-flight 555:568 arg1 spectrometry 575:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 511:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 7 theme -based 602:607 arg1 profiling 623:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 4 8 theme sialylation 984:994 arg1 levels 946:951 high levels 941:951 high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 941:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 8 theme sialylation 984:994 arg1 % 958:958 >95 % 954:958 >95 % 954:958 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 5 9 theme assisted 1097:1104 arg1 analysis 1087:1094 MS/MS structural analysis 1070:1094 MS/MS structural analysis 1070:1094 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 10 link α2-6-linked 1301:1311 arg1 NeuGc 1313:1317 α2-6-linked NeuGc 1301:1317 α2-6-linked NeuGc 1301:1317 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 11 theme linkage-specific 1109:1124 arg1 derivatization 1126:1139 linkage-specific derivatization 1109:1139 linkage-specific derivatization of sialic acids 1109:1155 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 1 12 dep initiation 236:245 arg1 the 232:234 the 232:234 the 232:234 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 6 13 link α2-3-linked 1461:1471 arg1 NeuGc 1473:1477 α2-3-linked NeuGc 1461:1477 α2-3-linked NeuGc 1461:1477 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 5 14 theme MS/MS 1070:1074 arg1 analysis 1087:1094 MS/MS structural analysis 1070:1094 MS/MS structural analysis 1070:1094 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 4 15 gly sialylation 984:994 arg1 fucosylation 1039:1050 core fucosylation 1034:1050 core fucosylation 1034:1050 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 16 theme galactosylation 964:978 arg1 levels 946:951 high levels 941:951 high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 941:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 16 theme galactosylation 964:978 arg1 % 958:958 >95 % 954:958 >95 % 954:958 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 5 17 gly disialylated 1220:1231 arg1 " 1265:1265 "branching sialylation" 1243:1265 "branching sialylation" 1243:1265 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 17 gly disialylated 1220:1231 arg1 antennae 1233:1240 disialylated antennae 1220:1240 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 7 18 theme biological 1772:1781 arg1 mechanisms 1783:1792 the biological mechanisms 1768:1792 the biological mechanisms involved 1768:1801 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 4 19 theme degree 1024:1029 arg1 levels 946:951 high levels 941:951 high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 941:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 19 theme degree 1024:1029 arg1 % 958:958 >95 % 954:958 >95 % 954:958 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 1 20 theme used 158:161 arg1 model 163:167 a widely used model 149:167 a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 149:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 1 20 theme used 158:161 arg1 peritonitis 134:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 4 21 theme core 1034:1037 arg1 fucosylation 1039:1050 core fucosylation 1034:1050 core fucosylation 1034:1050 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 0 22 theme fluid 80:84 arg1 N-glycosylation 86:100 plasma/peritoneal fluid N-glycosylation 62:100 plasma/peritoneal fluid N-glycosylation in mice 62:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 7 23 from changes 1636:1642 arg1 glycosylation 1647:1659 glycosylation 1647:1659 glycosylation 1647:1659 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 7 24 theme unexpected 1745:1754 arg1 velocity 1756:1763 an unexpected velocity 1742:1763 an unexpected velocity of the biological mechanisms involved 1742:1801 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 5 25 theme α2-6-linked 1301:1311 arg1 NeuGc 1313:1317 α2-6-linked NeuGc 1301:1317 α2-6-linked NeuGc 1301:1317 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 4 26 theme glycosylation 839:851 arg1 patterns 853:860 highly similar glycosylation patterns 824:860 highly similar glycosylation patterns 824:860 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 5 27 theme sialylation 1254:1264 arg1 " 1265:1265 "branching sialylation" 1243:1265 "branching sialylation" 1243:1265 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 27 theme sialylation 1254:1264 arg1 antennae 1233:1240 disialylated antennae 1220:1240 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 28 theme acids 1151:1155 arg1 derivatization 1126:1139 linkage-specific derivatization 1109:1139 linkage-specific derivatization of sialic acids 1109:1155 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 0 29 gly N-glycosylation 86:100 arg1 mice 105:108 mice 105:108 mice 105:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 0 30 theme plasma/peritoneal 62:78 arg1 N-glycosylation 86:100 plasma/peritoneal fluid N-glycosylation 62:100 plasma/peritoneal fluid N-glycosylation in mice 62:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 0 31 theme Acute 0:4 arg1 inflammation 12:23 Acute phase inflammation 0:23 Acute phase inflammation 0:23 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 6 32 dep together 1427:1434 arg1 with 1436:1439 with 1436:1439 with 1436:1439 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 6 33 theme α2-3-linked 1461:1471 arg1 NeuGc 1473:1477 α2-3-linked NeuGc 1461:1477 α2-3-linked NeuGc 1461:1477 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 5 34 theme disialylated 1220:1231 arg1 " 1265:1265 "branching sialylation" 1243:1265 "branching sialylation" 1243:1265 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 34 theme disialylated 1220:1231 arg1 antennae 1233:1240 disialylated antennae 1220:1240 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 1 35 theme Murine 111:116 arg1 model 163:167 a widely used model 149:167 a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 149:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 1 35 theme Murine 111:116 arg1 peritonitis 134:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 3 36 theme zymosan-induced 693:707 arg1 peritonitis 709:719 zymosan-induced peritonitis 693:719 zymosan-induced peritonitis 693:719 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 4 37 theme diantennary 884:894 arg1 N-glycans 926:934 diantennary and triantennary complex type N-glycans 884:934 diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 884:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 5 38 theme terminal 1365:1372 arg1 motif 1374:1378 the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1336:1378 the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1336:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 39 theme O-acetylated 1183:1194 arg1 acids 1203:1207 O-acetylated sialic acids 1183:1207 O-acetylated sialic acids 1183:1207 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 2 40 theme important 460:468 arg1 role 470:473 an important role 457:473 an important role 457:473 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 2 41 theme inflammatory 482:493 arg1 response 495:502 the inflammatory response 478:502 the inflammatory response 478:502 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 4 42 theme triantennary 900:911 arg1 N-glycans 926:934 diantennary and triantennary complex type N-glycans 884:934 diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 884:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 0 43 theme rapid 45:49 arg1 changes 51:57 rapid changes 45:57 rapid changes in plasma/peritoneal fluid N-glycosylation in mice 45:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 5 44 theme acids 1203:1207 arg1 presence 1171:1178 the presence 1167:1178 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 44 theme acids 1203:1207 arg1 " 1265:1265 "branching sialylation" 1243:1265 "branching sialylation" 1243:1265 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 44 theme acids 1203:1207 arg1 antennae 1233:1240 disialylated antennae 1220:1240 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 3 45 gly N-glycosylation 724:738 arg1 plasma 749:754 mouse plasma 743:754 mouse plasma 743:754 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 45 gly N-glycosylation 724:738 arg1 fluid 771:775 peritoneal fluid 760:775 peritoneal fluid 760:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 46 theme matrix-assisted 511:525 arg1 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 46 theme matrix-assisted 511:525 arg1 spectrometry 575:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 511:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 47 from N-glycosylation 724:738 arg1 plasma 749:754 mouse plasma 743:754 mouse plasma 743:754 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 47 from N-glycosylation 724:738 arg1 fluid 771:775 peritoneal fluid 760:775 peritoneal fluid 760:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 48 theme peritoneal 760:769 arg1 fluid 771:775 peritoneal fluid 760:775 peritoneal fluid 760:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 6 49 theme " 1505:1505 arg1 increase 1444:1451 an increase 1441:1451 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 4 50 theme high 941:944 arg1 levels 946:951 high levels 941:951 high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 941:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 50 theme high 941:944 arg1 % 958:958 >95 % 954:958 >95 % 954:958 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 3 51 theme mass 570:573 arg1 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 51 theme mass 570:573 arg1 spectrometry 575:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 511:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 6 52 theme branching 1484:1492 arg1 " 1505:1505 "branching sialylation" 1483:1505 "branching sialylation" 1483:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 5 53 theme branching 1244:1252 arg1 " 1265:1265 "branching sialylation" 1243:1265 "branching sialylation" 1243:1265 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 53 theme branching 1244:1252 arg1 antennae 1233:1240 disialylated antennae 1220:1240 the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1167:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 2 54 from changes 356:362 arg1 glycosylation 367:379 glycosylation 367:379 glycosylation of proteins 367:391 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 5 55 attach presence 1289:1296 arg2 NeuGc 1313:1317 α2-6-linked NeuGc 1301:1317 α2-6-linked NeuGc 1301:1317 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 55 attach presence 1289:1296 arg1 GlcNAc 1326:1331 the GlcNAc 1322:1331 the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1322:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 3 56 theme desorption/ionization 533:553 arg1 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 56 theme desorption/ionization 533:553 arg1 spectrometry 575:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 511:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 4 57 theme complex 913:919 arg1 N-glycans 926:934 diantennary and triantennary complex type N-glycans 884:934 diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 884:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 5 58 from presence 1289:1296 arg1 GlcNAc 1326:1331 the GlcNAc 1322:1331 the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1322:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 1 59 theme molecular 186:194 arg1 events 209:214 the molecular and cellular events 182:214 the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 182:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 6 60 theme significant 1383:1393 arg1 decrease 1395:1402 A significant decrease 1381:1402 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 1 61 theme cellular 200:207 arg1 events 209:214 the molecular and cellular events 182:214 the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 182:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 0 62 from N-glycosylation 86:100 arg1 mice 105:108 mice 105:108 mice 105:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 6 63 theme mice 1538:1541 arg1 N-glycomes 1524:1533 N-glycomes 1524:1533 N-glycomes of mice challenged with zymosan 1524:1565 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 1 64 theme responsible 216:226 arg1 events 209:214 the molecular and cellular events 182:214 the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 182:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 3 65 theme glycosylation 609:621 arg1 profiling 623:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 1 66 theme zymosan-induced 118:132 arg1 model 163:167 a widely used model 149:167 a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation 149:292 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 1 66 theme zymosan-induced 118:132 arg1 peritonitis 134:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis 111:144 Murine zymosan-induced peritonitis is a widely used model for studying the molecular and cellular events responsible for the initiation, persistence and/or resolution of inflammation. 26924641 7 67 theme substantial 1624:1634 arg1 changes 1636:1642 substantial changes 1624:1642 substantial changes in glycosylation 1624:1659 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 5 68 theme NeuGc 1313:1317 arg1 presence 1289:1296 the presence 1285:1296 the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1285:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 69 theme structural 1076:1085 arg1 analysis 1087:1094 MS/MS structural analysis 1070:1094 MS/MS structural analysis 1070:1094 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 3 70 theme qualitative 654:664 arg1 effect 683:688 the qualitative and quantitative effect 650:688 the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid 650:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 4 71 dep galactosylation 964:978 arg1 NeuGc 1004:1008 NeuGc 1004:1008 mostly NeuGc 997:1008 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 7 72 theme mechanisms 1783:1792 arg1 velocity 1756:1763 an unexpected velocity 1742:1763 an unexpected velocity of the biological mechanisms involved 1742:1801 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 4 73 theme medium 1017:1022 arg1 degree 1024:1029 a medium degree 1015:1029 a medium degree of core fucosylation (30 %) 1015:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 73 theme medium 1017:1022 arg1 % 1056:1056 30 % 1053:1056 30 % 1053:1056 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 6 74 theme fucosylation 1414:1425 arg1 decrease 1395:1402 A significant decrease 1381:1402 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 6 75 theme control 1579:1585 arg1 mice 1587:1590 control mice 1579:1590 control mice injected with PBS 1579:1608 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 4 76 with N-glycans 926:934 arg1 levels 946:951 high levels 941:951 high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %) 941:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 76 with N-glycans 926:934 arg1 % 958:958 >95 % 954:958 >95 % 954:958 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 6 77 theme core 1408:1411 arg1 fucosylation 1414:1425 (core) fucosylation 1407:1425 (core) fucosylation 1407:1425 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 0 78 from changes 51:57 arg1 N-glycosylation 86:100 plasma/peritoneal fluid N-glycosylation 62:100 plasma/peritoneal fluid N-glycosylation in mice 62:108 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 5 79 theme motif 1374:1378 arg1 GlcNAc 1326:1331 the GlcNAc 1322:1331 the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1322:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 5 80 theme sialic 1144:1149 arg1 acids 1151:1155 sialic acids 1144:1155 sialic acids 1144:1155 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 4 81 theme similar 831:837 arg1 patterns 853:860 highly similar glycosylation patterns 824:860 highly similar glycosylation patterns 824:860 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 0 82 theme phase 6:10 arg1 inflammation 12:23 Acute phase inflammation 0:23 Acute phase inflammation 0:23 Acute phase inflammation is characterized by rapid changes in plasma/peritoneal fluid N-glycosylation in mice. 26924641 2 83 theme inflammation 438:449 arg1 inflammation 438:449 inflammation 438:449 inflammation 438:449 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 2 83 theme inflammation 438:449 arg1 site 430:433 the site 426:433 the site of inflammation 426:449 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 6 84 theme NeuGc 1473:1477 arg1 increase 1444:1451 an increase 1441:1451 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 7 85 theme peritonitis 1707:1717 arg1 induction 1694:1702 induction 1694:1702 induction of peritonitis 1694:1717 Importantly, substantial changes in glycosylation were already observed 12 h after induction of peritonitis, thereby demonstrating an unexpected velocity of the biological mechanisms involved. 26924641 5 86 theme NeuGcα2-3-Galβ1-3-GlcNAc 1340:1363 arg1 motif 1374:1378 the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1336:1378 the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif 1336:1378 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 2 87 theme proteins 384:391 arg1 glycosylation 367:379 glycosylation 367:379 glycosylation of proteins 367:391 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 4 88 theme fucosylation 1039:1050 arg1 degree 1024:1029 a medium degree 1015:1029 a medium degree of core fucosylation (30 %) 1015:1057 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 88 theme fucosylation 1039:1050 arg1 galactosylation 964:978 galactosylation 964:978 galactosylation 964:978 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 88 theme fucosylation 1039:1050 arg1 % 1056:1056 30 % 1053:1056 30 % 1053:1056 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 4 88 theme fucosylation 1039:1050 arg1 sialylation 984:994 sialylation 984:994 sialylation 984:994 Our results show that both N-glycomes exhibit highly similar glycosylation patterns, consisting mainly of diantennary and triantennary complex type N-glycans with high levels (>95 %) of galactosylation and sialylation (mostly NeuGc) and a medium degree of core fucosylation (30 %). 26924641 2 89 gly glycosylation 367:379 arg1 proteins 384:391 proteins 384:391 proteins 384:391 Among these events, it is becoming increasingly evident that changes in glycosylation of proteins, especially in the plasma and at the site of inflammation, play an important role in the inflammatory response. 26924641 3 90 theme quantitative 670:681 arg1 effect 683:688 the qualitative and quantitative effect 650:688 the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid 650:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 5 91 theme sialic 1196:1201 arg1 acids 1203:1207 O-acetylated sialic acids 1183:1207 O-acetylated sialic acids 1183:1207 Moreover, MS/MS structural analysis, assisted by linkage-specific derivatization of sialic acids, revealed the presence of O-acetylated sialic acids as well as disialylated antennae ("branching sialylation") characterized by the presence of α2-6-linked NeuGc on the GlcNAc of the NeuGcα2-3-Galβ1-3-GlcNAc terminal motif. 26924641 6 92 located observed 1512:1519 arg2 decrease 1395:1402 A significant decrease 1381:1402 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 6 92 located observed 1512:1519 arg1 N-glycomes 1524:1533 N-glycomes 1524:1533 N-glycomes of mice challenged with zymosan 1524:1565 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 6 92 located observed 1512:1519 arg2 increase 1444:1451 an increase 1441:1451 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" 1381:1505 A significant decrease of (core) fucosylation together with an increase of both α2-3-linked NeuGc and "branching sialylation" were observed in N-glycomes of mice challenged with zymosan, but not in control mice injected with PBS. 26924641 3 93 theme peritonitis 709:719 arg1 effect 683:688 the qualitative and quantitative effect 650:688 the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid 650:775 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 94 theme laser 527:531 arg1 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 MALDI-TOF-MS 589:600 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 26924641 3 94 theme laser 527:531 arg1 spectrometry 575:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 511:586 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling 511:631 Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)-based glycosylation profiling, we investigated the qualitative and quantitative effect of zymosan-induced peritonitis on N-glycosylation in mouse plasma and peritoneal fluid. 27038031 8 0 theme glycosylation 1119:1131 arg1 Asn563 1091:1096 Asn563 1091:1096 Asn563 1091:1096 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 0 theme glycosylation 1119:1131 arg1 site 1133:1136 a high mannose-rich glycosylation site 1099:1136 a high mannose-rich glycosylation site that locates in the center of the IgM polymer 1099:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 6 1 theme N-linked 893:900 arg1 sites 902:906 the five N-linked sites 884:906 the five N-linked sites on the IgM heavy chains 884:930 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 9 2 theme peptide 1624:1630 arg1 profiling 1632:1640 global glycan or deglycosylated peptide profiling 1592:1640 profiling 1632:1640 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 7 3 contain had 970:972 arg1 Asn395 959:964 Asn395 959:964 Asn395 959:964 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 7 3 contain had 970:972 arg1 Asn171 939:944 Asn171 939:944 Asn171 939:944 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 7 3 contain had 970:972 arg1 Asn332 947:952 Asn332 947:952 Asn332 947:952 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 7 3 contain had 970:972 arg2 glycans 1001:1007 predominantly complex type glycans 974:1007 predominantly complex type glycans 974:1007 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 2 4 theme heavy 277:281 arg1 monomer 289:295 each heavy chain monomer 272:295 each heavy chain monomer 272:295 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 5 from antibody 203:210 arg1 humans 215:220 humans 215:220 humans 215:220 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 1 6 theme important 115:123 arg1 role 125:128 an important role 112:128 an important role 112:128 Glycosylation is known to play an important role in IgG antibody structure and function. 27038031 4 7 theme culture 577:583 arg1 conditions 585:594 culture conditions 577:594 culture conditions 577:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 4 8 gly glycopeptides 688:700 arg2 glycopeptides 688:700 HILIC-enriched tryptic and GluC glycopeptides 656:700 glycopeptides 688:700 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 8 9 theme polymer 1176:1182 arg1 center 1158:1163 the center 1154:1163 the center of the IgM polymer 1154:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 4 10 theme GluC 683:686 arg1 glycopeptides 688:700 HILIC-enriched tryptic and GluC glycopeptides 656:700 glycopeptides 688:700 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 5 11 from occupancy 707:715 arg1 IgM 787:789 the pentameric and hexameric IgM 758:789 the pentameric and hexameric IgM 758:789 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 7 12 theme complex 988:994 arg1 glycans 1001:1007 predominantly complex type glycans 974:1007 predominantly complex type glycans 974:1007 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 9 13 with mass 1512:1515 arg1 structure 1533:1541 quaternary structure 1522:1541 quaternary structure 1522:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 6 14 theme IgM 915:917 arg1 chains 925:930 the IgM heavy chains 911:930 the IgM heavy chains 911:930 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 8 15 theme IgM 1258:1260 arg1 forms 1262:1266 both the pentameric and hexameric IgM forms 1224:1266 forms 1262:1266 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 16 theme structures 1323:1332 arg1 abundance 1299:1307 relative abundance 1290:1307 relative abundance of the glycan structures 1290:1332 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 2 17 theme N-glycosylation 247:261 arg1 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 1 18 theme IgG 133:135 arg1 structure 146:154 IgG antibody structure 133:154 IgG antibody structure 133:154 Glycosylation is known to play an important role in IgG antibody structure and function. 27038031 5 19 theme sites 749:753 arg1 able 796:799 able 796:799 able 796:799 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 5 19 theme sites 749:753 arg1 occupancy 707:715 The occupancy 703:715 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM 703:789 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 4 20 gly N-glycosylation 476:490 arg1 IgM 532:534 recombinant pentameric and hexameric IgM 495:534 recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions 495:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 4 20 gly N-glycosylation 476:490 arg1 study 465:469 this study 460:469 this study 460:469 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 9 21 theme protein 1486:1492 arg1 site-heterogeneity 1442:1459 the site-heterogeneity 1438:1459 the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure 1438:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 9 22 gly site-heterogeneity 1442:1459 arg1 protein 1486:1492 a highly glycosylated protein 1464:1492 a highly glycosylated protein of high molecular mass with quaternary structure 1464:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 9 23 theme high 1497:1500 arg1 mass 1512:1515 high molecular mass 1497:1515 high molecular mass with quaternary structure 1497:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 3 24 theme connecting 333:342 arg1 J-chain 366:372 a connecting singly N-glycosylated J-chain 331:372 a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) 331:413 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 9 25 theme mass 1512:1515 arg1 protein 1486:1492 a highly glycosylated protein 1464:1492 a highly glycosylated protein of high molecular mass with quaternary structure 1464:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 9 26 gly glycosylated 1473:1484 arg1 protein 1486:1492 a highly glycosylated protein 1464:1492 a highly glycosylated protein of high molecular mass with quaternary structure 1464:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 5 27 theme putative 724:731 arg1 sites 749:753 all putative N-glycosylation sites 720:753 all putative N-glycosylation sites 720:753 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 9 28 theme quaternary 1522:1531 arg1 structure 1533:1541 quaternary structure 1522:1541 quaternary structure 1522:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 3 29 theme glycosylation 394:406 arg1 sites 408:412 51 glycosylation sites 391:412 51 glycosylation sites 391:412 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 0 30 theme Site-Specific 0:12 arg1 N-Glycosylation 14:28 Site-Specific N-Glycosylation 0:28 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 0:79 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 4 31 theme recombinant 495:505 arg1 IgM 532:534 recombinant pentameric and hexameric IgM 495:534 recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions 495:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 3 32 gly glycosylation 435:447 arg2 60 432:433 60 432:433 60 432:433 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 3 32 gly glycosylation 435:447 arg2 hexamer 423:429 a hexamer 421:429 a hexamer (60 glycosylation sites) 421:454 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 3 32 gly glycosylation 435:447 arg2 sites 449:453 60 glycosylation sites 432:453 60 glycosylation sites 432:453 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 1 33 theme antibody 137:144 arg1 structure 146:154 IgG antibody structure 133:154 IgG antibody structure 133:154 Glycosylation is known to play an important role in IgG antibody structure and function. 27038031 10 34 theme Graphical 1643:1651 arg1 Abstract 1653:1660 Graphical Abstract 1643:1660 Graphical Abstract 1643:1660 Graphical Abstract ᅟ. 27038031 0 35 theme Pentameric 45:54 arg1 N-Glycosylation 14:28 Site-Specific N-Glycosylation 0:28 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 0:79 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 0 36 gly N-Glycosylation 14:28 arg1 IgM 76:78 Hexameric Human IgM 60:78 Hexameric Human IgM 60:78 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 0 36 gly N-Glycosylation 14:28 arg1 Pentameric 45:54 Recombinant Pentameric 33:54 Recombinant Pentameric 33:54 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 0 37 theme IgM 76:78 arg1 N-Glycosylation 14:28 Site-Specific N-Glycosylation 0:28 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 0:79 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 0 38 theme Hexameric 60:68 arg1 IgM 76:78 Hexameric Human IgM 60:78 Hexameric Human IgM 60:78 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 6 39 theme glycosylation 828:840 arg1 differences 842:852 Distinct glycosylation differences 819:852 Distinct glycosylation differences 819:852 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 8 40 dep forms 1262:1266 arg1 both 1224:1227 both 1224:1227 both 1224:1227 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 3 41 dep connecting 333:342 arg1 N-glycosylated 351:364 N-glycosylated 351:364 N-glycosylated 351:364 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 2 42 theme known 197:201 arg1 IgM 180:182 Polymeric IgM 170:182 Polymeric IgM 170:182 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 42 theme known 197:201 arg1 antibody 203:210 the largest known antibody 185:210 the largest known antibody in humans 185:220 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 8 43 gly glycosylation 1119:1131 arg2 Asn563 1091:1096 Asn563 1091:1096 Asn563 1091:1096 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 43 gly glycosylation 1119:1131 arg2 site 1133:1136 a high mannose-rich glycosylation site 1099:1136 a high mannose-rich glycosylation site that locates in the center of the IgM polymer 1099:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 43 gly glycosylation 1119:1131 arg1 center 1158:1163 the center 1154:1163 the center of the IgM polymer 1154:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 44 dep pentamer 1346:1353 arg1 the 1342:1344 the 1342:1344 the 1342:1344 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 4 45 theme same 552:555 arg1 type 568:571 the same human cell type 548:571 the same human cell type 548:571 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 6 46 from sites 902:906 arg1 chains 925:930 the IgM heavy chains 911:930 the IgM heavy chains 911:930 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 8 47 from difference 1276:1285 arg1 abundance 1299:1307 relative abundance 1290:1307 relative abundance of the glycan structures 1290:1332 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 0 48 theme Human 70:74 arg1 IgM 76:78 Hexameric Human IgM 60:78 Hexameric Human IgM 60:78 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 4 49 theme cell 563:566 arg1 type 568:571 the same human cell type 548:571 the same human cell type 548:571 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 8 50 theme mannose-rich 1106:1117 arg1 Asn563 1091:1096 Asn563 1091:1096 Asn563 1091:1096 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 50 theme mannose-rich 1106:1117 arg1 site 1133:1136 a high mannose-rich glycosylation site 1099:1136 a high mannose-rich glycosylation site that locates in the center of the IgM polymer 1099:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 9 51 theme deglycosylated 1609:1622 arg1 profiling 1632:1640 global glycan or deglycosylated peptide profiling 1592:1640 profiling 1632:1640 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 4 52 theme IgM 532:534 arg1 N-glycosylation 476:490 the N-glycosylation 472:490 the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions 472:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 2 53 theme chain 283:287 arg1 monomer 289:295 each heavy chain monomer 272:295 each heavy chain monomer 272:295 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 54 gly N-glycosylation 247:261 arg2 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 54 gly N-glycosylation 247:261 arg2 five 232:235 five 232:235 five 232:235 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 3 55 with J-chain 366:372 arg1 total 382:386 a total 380:386 a total of 51 glycosylation sites 380:412 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 7 56 theme glycan 1025:1030 arg1 branching 1032:1040 glycan branching 1025:1040 glycan branching 1025:1040 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 8 57 theme IgM 1172:1174 arg1 polymer 1176:1182 the IgM polymer 1168:1182 the IgM polymer 1168:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 7 58 theme type 996:999 arg1 glycans 1001:1007 predominantly complex type glycans 974:1007 predominantly complex type glycans 974:1007 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 9 59 theme site-heterogeneity 1442:1459 arg1 characterization 1418:1433 characterization 1418:1433 characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure 1418:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 6 60 theme heavy 919:923 arg1 chains 925:930 the IgM heavy chains 911:930 the IgM heavy chains 911:930 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 8 61 theme pentameric 1233:1242 arg1 forms 1262:1266 both the pentameric and hexameric IgM forms 1224:1266 forms 1262:1266 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 62 theme hexameric 1248:1256 arg1 forms 1262:1266 both the pentameric and hexameric IgM forms 1224:1266 forms 1262:1266 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 5 63 theme N-glycosylation 733:747 arg1 sites 749:753 all putative N-glycosylation sites 720:753 all putative N-glycosylation sites 720:753 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 2 64 theme potential 237:245 arg1 sites 263:267 five potential N-glycosylation sites 232:267 five potential N-glycosylation sites 232:267 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 9 65 theme glycosylated 1473:1484 arg1 protein 1486:1492 a highly glycosylated protein 1464:1492 a highly glycosylated protein of high molecular mass with quaternary structure 1464:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 3 66 with pentamer 317:324 arg1 J-chain 366:372 a connecting singly N-glycosylated J-chain 331:372 a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) 331:413 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 9 67 theme molecular 1502:1510 arg1 mass 1512:1515 high molecular mass 1497:1515 high molecular mass with quaternary structure 1497:1541 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 5 68 gly N-glycosylation 733:747 arg2 sites 749:753 all putative N-glycosylation sites 720:753 all putative N-glycosylation sites 720:753 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 8 69 theme relative 1290:1297 arg1 abundance 1299:1307 relative abundance 1290:1307 relative abundance of the glycan structures 1290:1332 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 6 70 link N-linked 893:900 arg1 sites 902:906 the five N-linked sites 884:906 the five N-linked sites on the IgM heavy chains 884:930 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 4 71 theme pentameric 507:516 arg1 IgM 532:534 recombinant pentameric and hexameric IgM 495:534 recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions 495:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 5 72 gly occupancy 707:715 arg2 sites 749:753 all putative N-glycosylation sites 720:753 all putative N-glycosylation sites 720:753 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 3 73 theme sites 408:412 arg1 total 382:386 a total 380:386 a total of 51 glycosylation sites 380:412 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 0 74 theme Recombinant 33:43 arg1 Pentameric 45:54 Recombinant Pentameric 33:54 Recombinant Pentameric 33:54 Site-Specific N-Glycosylation of Recombinant Pentameric and Hexameric Human IgM. 27038031 4 75 theme hexameric 522:530 arg1 IgM 532:534 recombinant pentameric and hexameric IgM 495:534 recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions 495:594 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 8 76 theme glycan 1316:1321 arg1 structures 1323:1332 the glycan structures 1312:1332 the glycan structures 1312:1332 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 5 77 theme pentameric 762:771 arg1 IgM 787:789 the pentameric and hexameric IgM 758:789 the pentameric and hexameric IgM 758:789 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 2 78 theme Polymeric 170:178 arg1 IgM 180:182 Polymeric IgM 170:182 Polymeric IgM 170:182 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 78 theme Polymeric 170:178 arg1 antibody 203:210 the largest known antibody 185:210 the largest known antibody in humans 185:220 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 5 79 theme hexameric 777:785 arg1 IgM 787:789 the pentameric and hexameric IgM 758:789 the pentameric and hexameric IgM 758:789 The occupancy of all putative N-glycosylation sites on the pentameric and hexameric IgM were able to be determined. 27038031 6 80 theme Distinct 819:826 arg1 differences 842:852 Distinct glycosylation differences 819:852 Distinct glycosylation differences 819:852 Distinct glycosylation differences were observed between each of the five N-linked sites on the IgM heavy chains. 27038031 4 81 theme human 557:561 arg1 type 568:571 the same human cell type 548:571 the same human cell type 548:571 In this study, the N-glycosylation of recombinant pentameric and hexameric IgM produced by the same human cell type and culture conditions was site-specifically profiled by RP-LC-CID/ETD-MS/MS using HILIC-enriched tryptic and GluC glycopeptides. 27038031 2 82 theme largest 189:195 arg1 IgM 180:182 Polymeric IgM 170:182 Polymeric IgM 170:182 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 2 82 theme largest 189:195 arg1 antibody 203:210 the largest known antibody 185:210 the largest known antibody in humans 185:220 Polymeric IgM, the largest known antibody in humans, displays five potential N-glycosylation sites on each heavy chain monomer. 27038031 3 83 theme glycosylation 435:447 arg1 sites 449:453 60 glycosylation sites 432:453 60 glycosylation sites 432:453 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 3 83 theme glycosylation 435:447 arg1 hexamer 423:429 a hexamer 421:429 a hexamer (60 glycosylation sites) 421:454 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 8 84 theme high 1101:1104 arg1 Asn563 1091:1096 Asn563 1091:1096 Asn563 1091:1096 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 8 84 theme high 1101:1104 arg1 site 1133:1136 a high mannose-rich glycosylation site 1099:1136 a high mannose-rich glycosylation site that locates in the center of the IgM polymer 1099:1182 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 9 85 theme global 1592:1597 arg1 glycan 1599:1604 global glycan or deglycosylated peptide profiling 1592:1640 glycan 1599:1604 This study highlights the information obtained by characterization of the site-heterogeneity of a highly glycosylated protein of high molecular mass with quaternary structure, revealing differences that would not be seen by global glycan or deglycosylated peptide profiling. 27038031 7 86 from differences 1010:1020 arg1 branching 1032:1040 glycan branching 1025:1040 glycan branching 1025:1040 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 7 86 from differences 1010:1020 arg1 sialylation 1046:1056 sialylation 1046:1056 sialylation 1046:1056 While Asn171, Asn332, and Asn395 all had predominantly complex type glycans, differences in glycan branching and sialylation were observed between the sites. 27038031 8 87 with % 1210:1210 arg1 difference 1276:1285 a difference 1274:1285 a difference in relative abundance of the glycan structures between the pentamer and hexamer 1274:1365 Asn563, a high mannose-rich glycosylation site that locates in the center of the IgM polymer, was only approximately 60% occupied in both the pentameric and hexameric IgM forms, with a difference in relative abundance of the glycan structures between the pentamer and hexamer. 27038031 3 88 gly glycosylation 394:406 arg2 51 391:392 51 391:392 51 391:392 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 27038031 3 88 gly glycosylation 394:406 arg2 sites 408:412 51 glycosylation sites 391:412 51 glycosylation sites 391:412 IgM can exist as a pentamer with a connecting singly N-glycosylated J-chain (with a total of 51 glycosylation sites) or as a hexamer (60 glycosylation sites). 29273683 8 0 theme higher 1148:1153 arg1 affinities 1182:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 3 1 theme steroid-binding 247:261 arg1 activity 263:270 its steroid-binding activity 243:270 its steroid-binding activity 243:270 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 2 from sites 303:307 arg1 RCLs 339:342 RCLs 339:342 RCLs 339:342 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 2 from sites 303:307 arg1 loops 332:336 the reactive center loops 312:336 the reactive center loops (RCLs) of human and rat CBGs 312:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 7 3 theme human 736:740 arg1 CBG 742:744 human CBG 736:744 human CBG 736:744 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 4 4 theme CBG 400:402 arg1 RCL 387:389 the RCL 383:389 the RCL of human CBG 383:402 Proteolysis of the RCL of human CBG causes a structural change that disrupts steroid binding. 29273683 9 5 from presence 1298:1305 arg1 position 1356:1363 this conserved position 1341:1363 this conserved position 1341:1363 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 2 6 theme rat 164:166 arg1 CBGs 168:171 Human and rat CBGs 154:171 Human and rat CBGs 154:171 Human and rat CBGs have six N-glycosylation sites. 29273683 5 7 theme conserved 492:500 arg1 sites 518:522 conserved N-glycosylation sites 492:522 conserved N-glycosylation sites 492:522 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 11 8 from glycosylation 1634:1646 arg1 RCL 1663:1665 the RCL 1659:1665 the RCL 1659:1665 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 8 9 theme N-acetylglucosaminyltransferase-I-deficient 1002:1044 arg1 cells 1051:1055 N-acetylglucosaminyltransferase-I-deficient Lec1 cells 1002:1055 N-acetylglucosaminyltransferase-I-deficient Lec1 cells 1002:1055 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 10 theme CHO-S 1275:1279 arg1 cells 1281:1285 glycosylation-competent CHO-S cells 1251:1285 glycosylation-competent CHO-S cells 1251:1285 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 5 11 from N230 549:552 arg1 CBG 561:563 rat CBG 557:563 rat CBG 557:563 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 5 11 from N230 549:552 arg1 mutations 479:487 mutations 479:487 mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG 479:563 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 3 12 theme N-glycosylation 287:301 arg1 sites 303:307 N-glycosylation sites 287:307 N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs 287:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 11 13 gly glycosylation 1634:1646 arg1 RCL 1663:1665 the RCL 1659:1665 the RCL 1659:1665 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 11 13 gly glycosylation 1634:1646 arg1 N347 1651:1654 N347 1651:1654 N347 1651:1654 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 5 14 theme human 535:539 arg1 CBG 541:543 human CBG 535:543 human CBG 535:543 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 8 15 theme Lec2 1237:1240 arg1 cells 1242:1246 sialylation-deficient Lec2 cells 1215:1246 sialylation-deficient Lec2 cells 1215:1246 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 9 16 theme conserved 1346:1354 arg1 position 1356:1363 this conserved position 1341:1363 this conserved position 1341:1363 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 1 17 theme glucocorticoids 137:151 arg1 globulin 99:106 Corticosteroid-binding globulin 76:106 Corticosteroid-binding globulin (CBG) 76:112 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 1 17 theme glucocorticoids 137:151 arg1 carrier 126:132 a plasma carrier 117:132 a plasma carrier of glucocorticoids 117:151 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 3 18 theme reactive 316:323 arg1 RCLs 339:342 RCLs 339:342 RCLs 339:342 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 18 theme reactive 316:323 arg1 loops 332:336 the reactive center loops 312:336 the reactive center loops (RCLs) of human and rat CBGs 312:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 5 19 theme sites 518:522 arg1 mutations 479:487 mutations 479:487 mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG 479:563 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 8 20 gly glycosylation 1115:1127 arg2 site 1129:1132 only one glycosylation site 1106:1132 only one glycosylation site 1106:1132 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 20 gly glycosylation 1115:1127 arg2 one 1111:1113 one 1111:1113 one 1111:1113 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 6 21 dep human 652:656 arg1 activities 686:695 CBG steroid-binding activities 666:695 CBG steroid-binding activities 666:695 Inhibiting glycosylation by tunicamycin also markedly reduced human and rat CBG steroid-binding activities. 29273683 8 22 theme human 1058:1062 arg1 CBGs 1072:1075 human and rat CBGs 1058:1075 human and rat CBGs 1058:1075 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 5 23 theme steroid 573:579 arg1 binding 581:587 steroid binding 573:587 steroid binding 573:587 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 2 24 contain have 173:176 arg1 CBGs 168:171 Human and rat CBGs 154:171 Human and rat CBGs 154:171 Human and rat CBGs have six N-glycosylation sites. 29273683 2 24 contain have 173:176 arg2 sites 198:202 six N-glycosylation sites 178:202 six N-glycosylation sites 178:202 Human and rat CBGs have six N-glycosylation sites. 29273683 3 25 theme CBGs 362:365 arg1 RCLs 339:342 RCLs 339:342 RCLs 339:342 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 25 theme CBGs 362:365 arg1 loops 332:336 the reactive center loops 312:336 the reactive center loops (RCLs) of human and rat CBGs 312:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 10 26 theme neutrophil 1443:1452 arg1 elastase 1454:1461 neutrophil elastase 1443:1461 neutrophil elastase 1443:1461 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 2 27 gly N-glycosylation 182:196 arg2 six 178:180 six 178:180 six 178:180 Human and rat CBGs have six N-glycosylation sites. 29273683 2 27 gly N-glycosylation 182:196 arg2 sites 198:202 six N-glycosylation sites 178:202 six N-glycosylation sites 178:202 Human and rat CBGs have six N-glycosylation sites. 29273683 8 28 theme rat 1068:1070 arg1 CBGs 1072:1075 human and rat CBGs 1058:1075 human and rat CBGs 1058:1075 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 11 29 from sites 1711:1715 arg1 N-glycans 1692:1700 N-glycans 1692:1700 N-glycans at other sites 1692:1715 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 8 30 dep higher 1148:1153 arg1 fold 1160:1163 2-4 fold 1156:1163 2-4 fold 1156:1163 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 7 31 gly glycosylated 723:734 arg1 CBG 742:744 human CBG 736:744 human CBG 736:744 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 9 32 theme N-glycan 1329:1336 arg1 composition 1311:1321 composition 1311:1321 composition 1311:1321 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 9 32 theme N-glycan 1329:1336 arg1 presence 1298:1305 presence 1298:1305 presence 1298:1305 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 7 33 theme N238 949:952 arg1 deamidation 934:944 deamidation 934:944 deamidation of N238 934:952 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 3 34 theme CBG 228:230 arg1 Glycosylation 205:217 Glycosylation 205:217 Glycosylation of human CBG 205:230 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 0 35 theme Functional 0:9 arg1 implications 11:22 Functional implications 0:22 Functional implications of corticosteroid-binding globulin N-glycosylation. 0:74 Functional implications of corticosteroid-binding globulin N-glycosylation. 29273683 7 36 theme H-reduced 800:808 arg1 affinity 826:833 Endo H-reduced steroid-binding affinity 795:833 Endo H-reduced steroid-binding affinity 795:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 7 37 with Deglycosylation 698:712 arg1 N-glycan 773:780 only one N-glycan 764:780 only one N-glycan at N238 with Endo H-reduced steroid-binding affinity 764:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 8 38 theme CBG 1090:1092 arg1 mutant 1094:1099 a human CBG mutant 1082:1099 a human CBG mutant with only one glycosylation site at N238 1082:1140 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 9 39 dep presence 1298:1305 arg1 the 1294:1296 the 1294:1296 the 1294:1296 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 0 40 theme globulin 50:57 arg1 N-glycosylation 59:73 corticosteroid-binding globulin N-glycosylation 27:73 corticosteroid-binding globulin N-glycosylation 27:73 Functional implications of corticosteroid-binding globulin N-glycosylation. 29273683 11 41 theme other 1705:1709 arg1 sites 1711:1715 other sites 1705:1715 other sites 1705:1715 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 9 42 theme steroid 1394:1400 arg1 binding 1402:1408 the steroid binding 1390:1408 the steroid binding of CBG 1390:1415 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 8 43 with mutant 1094:1099 arg1 site 1129:1132 only one glycosylation site 1106:1132 only one glycosylation site 1106:1132 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 7 44 theme F-mediated 849:858 arg1 deglycosylation 860:874 PNGase F-mediated deglycosylation 842:874 PNGase F-mediated deglycosylation 842:874 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 11 45 theme N347 1651:1654 arg1 glycosylation 1634:1646 glycosylation 1634:1646 glycosylation of N347 in the RCL 1634:1665 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 7 46 theme human 749:753 arg1 CBG 755:757 human CBG 749:757 human CBG 749:757 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 1 47 theme Corticosteroid-binding 76:97 arg1 CBG 109:111 CBG 109:111 CBG 109:111 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 1 47 theme Corticosteroid-binding 76:97 arg1 globulin 99:106 Corticosteroid-binding globulin 76:106 Corticosteroid-binding globulin (CBG) 76:112 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 1 47 theme Corticosteroid-binding 76:97 arg1 carrier 126:132 a plasma carrier 117:132 a plasma carrier of glucocorticoids 117:151 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 7 48 theme steroid 902:908 arg1 binding 910:916 steroid binding 902:916 steroid binding 902:916 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 7 49 theme CBG 742:744 arg1 Deglycosylation 698:712 Deglycosylation 698:712 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity 698:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 4 50 theme human 394:398 arg1 CBG 400:402 human CBG 394:402 human CBG 394:402 Proteolysis of the RCL of human CBG causes a structural change that disrupts steroid binding. 29273683 7 51 gly Deglycosylation 698:712 arg1 CBG 742:744 human CBG 736:744 human CBG 736:744 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 7 51 gly Deglycosylation 698:712 arg1 CBG 755:757 human CBG 749:757 human CBG 749:757 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 10 52 theme protease 1603:1610 arg1 LasB 1612:1615 the Pseudomonas aeruginosa protease LasB 1576:1615 the Pseudomonas aeruginosa protease LasB 1576:1615 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 7 53 theme glycosylated 723:734 arg1 CBG 742:744 human CBG 736:744 human CBG 736:744 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 7 54 from N238 785:788 arg1 N-glycan 773:780 only one N-glycan 764:780 only one N-glycan at N238 with Endo H-reduced steroid-binding affinity 764:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 8 55 with CBGs 1072:1075 arg1 site 1129:1132 only one glycosylation site 1106:1132 only one glycosylation site 1106:1132 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 10 56 theme Pseudomonas 1580:1590 arg1 LasB 1612:1615 the Pseudomonas aeruginosa protease LasB 1576:1615 the Pseudomonas aeruginosa protease LasB 1576:1615 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 7 57 with N238 785:788 arg1 affinity 826:833 Endo H-reduced steroid-binding affinity 795:833 Endo H-reduced steroid-binding affinity 795:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 5 58 from N238 527:530 arg1 CBG 541:543 human CBG 535:543 human CBG 535:543 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 5 58 from N238 527:530 arg1 mutations 479:487 mutations 479:487 mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG 479:563 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 3 59 gly N-glycosylation 287:301 arg2 sites 303:307 N-glycosylation sites 287:307 N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs 287:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 8 60 theme Lec1 1046:1049 arg1 cells 1051:1055 N-acetylglucosaminyltransferase-I-deficient Lec1 cells 1002:1055 N-acetylglucosaminyltransferase-I-deficient Lec1 cells 1002:1055 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 7 61 dep indicating 886:895 arg1 does 876:879 does 876:879 does not 876:883 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 8 62 contain have 1143:1146 arg1 CBGs 1072:1075 human and rat CBGs 1058:1075 human and rat CBGs 1058:1075 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 62 contain have 1143:1146 arg1 mutant 1094:1099 a human CBG mutant 1082:1099 a human CBG mutant with only one glycosylation site at N238 1082:1140 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 62 contain have 1143:1146 arg2 affinities 1182:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 5 63 theme N-glycosylation 502:516 arg1 sites 518:522 conserved N-glycosylation sites 492:522 conserved N-glycosylation sites 492:522 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 8 64 theme steroid-binding 1166:1180 arg1 affinities 1182:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 higher (2-4 fold) steroid-binding affinities 1148:1191 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 4 65 theme RCL 387:389 arg1 Proteolysis 368:378 Proteolysis 368:378 Proteolysis of the RCL of human CBG 368:402 Proteolysis of the RCL of human CBG causes a structural change that disrupts steroid binding. 29273683 8 66 theme glycosylation-competent 1251:1273 arg1 cells 1281:1285 glycosylation-competent CHO-S cells 1251:1285 glycosylation-competent CHO-S cells 1251:1285 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 10 67 dep Pseudomonas 1580:1590 arg1 aeruginosa 1592:1601 aeruginosa 1592:1601 aeruginosa 1592:1601 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 7 68 theme PNGase 842:847 arg1 deglycosylation 860:874 PNGase F-mediated deglycosylation 842:874 PNGase F-mediated deglycosylation 842:874 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 4 69 theme steroid 445:451 arg1 binding 453:459 steroid binding 445:459 steroid binding 445:459 Proteolysis of the RCL of human CBG causes a structural change that disrupts steroid binding. 29273683 9 70 from composition 1311:1321 arg1 position 1356:1363 this conserved position 1341:1363 this conserved position 1341:1363 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 10 71 dep does 1555:1558 arg1 than 1550:1553 than 1550:1553 than 1550:1553 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 8 72 theme sialylation-deficient 1215:1235 arg1 cells 1242:1246 sialylation-deficient Lec2 cells 1215:1246 sialylation-deficient Lec2 cells 1215:1246 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 4 73 theme structural 413:422 arg1 change 424:429 a structural change 411:429 a structural change that disrupts steroid binding 411:459 Proteolysis of the RCL of human CBG causes a structural change that disrupts steroid binding. 29273683 3 74 theme center 325:330 arg1 RCLs 339:342 RCLs 339:342 RCLs 339:342 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 74 theme center 325:330 arg1 loops 332:336 the reactive center loops 312:336 the reactive center loops (RCLs) of human and rat CBGs 312:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 75 gly Glycosylation 205:217 arg1 CBG 228:230 human CBG 222:230 human CBG 222:230 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 76 theme human 348:352 arg1 CBGs 362:365 human and rat CBGs 348:365 human and rat CBGs 348:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 11 77 theme neutrophil 1749:1758 arg1 elastase 1760:1767 neutrophil elastase 1749:1767 neutrophil elastase 1749:1767 Moreover, while glycosylation of N347 in the RCL limits these activities, N-glycans at other sites also appear to protect CBG from neutrophil elastase or chymotrypsin. 29273683 6 78 theme steroid-binding 670:684 arg1 activities 686:695 CBG steroid-binding activities 666:695 CBG steroid-binding activities 666:695 Inhibiting glycosylation by tunicamycin also markedly reduced human and rat CBG steroid-binding activities. 29273683 5 79 gly N-glycosylation 502:516 arg2 sites 518:522 conserved N-glycosylation sites 492:522 conserved N-glycosylation sites 492:522 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 9 80 attach presence 1298:1305 arg1 position 1356:1363 this conserved position 1341:1363 this conserved position 1341:1363 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 9 80 attach presence 1298:1305 arg2 N-glycan 1329:1336 an N-glycan 1326:1336 an N-glycan 1326:1336 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 29273683 3 81 theme rat 358:360 arg1 CBGs 362:365 human and rat CBGs 348:365 human and rat CBGs 348:365 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 3 82 theme human 222:226 arg1 CBG 228:230 human CBG 222:230 human CBG 222:230 Glycosylation of human CBG influences its steroid-binding activity, and there are N-glycosylation sites in the reactive center loops (RCLs) of human and rat CBGs. 29273683 8 83 theme human 1084:1088 arg1 mutant 1094:1099 a human CBG mutant 1082:1099 a human CBG mutant with only one glycosylation site at N238 1082:1140 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 5 84 theme rat 557:559 arg1 CBG 561:563 rat CBG 557:563 rat CBG 557:563 We now show that mutations of conserved N-glycosylation sites at N238 in human CBG and N230 in rat CBG disrupt steroid binding. 29273683 10 85 theme human 1482:1486 arg1 CBG 1488:1490 human CBG 1482:1490 human CBG 1482:1490 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 0 86 theme corticosteroid-binding 27:48 arg1 N-glycosylation 59:73 corticosteroid-binding globulin N-glycosylation 27:73 corticosteroid-binding globulin N-glycosylation 27:73 Functional implications of corticosteroid-binding globulin N-glycosylation. 29273683 7 87 theme steroid-binding 810:824 arg1 affinity 826:833 Endo H-reduced steroid-binding affinity 795:833 Endo H-reduced steroid-binding affinity 795:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 1 88 theme plasma 119:124 arg1 globulin 99:106 Corticosteroid-binding globulin 76:106 Corticosteroid-binding globulin (CBG) 76:112 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 1 88 theme plasma 119:124 arg1 carrier 126:132 a plasma carrier 117:132 a plasma carrier of glucocorticoids 117:151 Corticosteroid-binding globulin (CBG) is a plasma carrier of glucocorticoids. 29273683 6 89 theme CBG 666:668 arg1 activities 686:695 CBG steroid-binding activities 666:695 CBG steroid-binding activities 666:695 Inhibiting glycosylation by tunicamycin also markedly reduced human and rat CBG steroid-binding activities. 29273683 0 90 theme N-glycosylation 59:73 arg1 implications 11:22 Functional implications 0:22 Functional implications of corticosteroid-binding globulin N-glycosylation. 0:74 Functional implications of corticosteroid-binding globulin N-glycosylation. 29273683 7 91 theme Endo 795:798 arg1 affinity 826:833 Endo H-reduced steroid-binding affinity 795:833 Endo H-reduced steroid-binding affinity 795:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 10 92 theme CBG 1488:1490 arg1 RCL 1475:1477 the RCL 1471:1477 the RCL of human CBG 1471:1490 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 2 93 theme N-glycosylation 182:196 arg1 sites 198:202 six N-glycosylation sites 178:202 six N-glycosylation sites 178:202 Human and rat CBGs have six N-glycosylation sites. 29273683 8 94 theme glycosylation 1115:1127 arg1 site 1129:1132 only one glycosylation site 1106:1132 only one glycosylation site 1106:1132 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 95 from N238 1137:1140 arg1 CBGs 1072:1075 human and rat CBGs 1058:1075 human and rat CBGs 1058:1075 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 8 95 from N238 1137:1140 arg1 mutant 1094:1099 a human CBG mutant 1082:1099 a human CBG mutant with only one glycosylation site at N238 1082:1140 When expressed in N-acetylglucosaminyltransferase-I-deficient Lec1 cells, human and rat CBGs, and a human CBG mutant with only one glycosylation site at N238, have higher (2-4 fold) steroid-binding affinities than when produced by sialylation-deficient Lec2 cells or glycosylation-competent CHO-S cells. 29273683 10 96 theme steroid-binding 1508:1522 arg1 capacity 1524:1531 its steroid-binding capacity 1504:1531 its steroid-binding capacity 1504:1531 We also demonstrate that neutrophil elastase cleaves the RCL of human CBG and reduces its steroid-binding capacity more efficiently than does chymotrypsin or the Pseudomonas aeruginosa protease LasB. 29273683 2 97 theme Human 154:158 arg1 CBGs 168:171 Human and rat CBGs 154:171 Human and rat CBGs 154:171 Human and rat CBGs have six N-glycosylation sites. 29273683 7 98 theme CBG 755:757 arg1 Deglycosylation 698:712 Deglycosylation 698:712 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity 698:833 Deglycosylation of fully glycosylated human CBG or human CBG with only one N-glycan at N238 with Endo H-reduced steroid-binding affinity, while PNGase F-mediated deglycosylation does not, indicating that steroid binding is preserved by deamidation of N238 when its N-glycan is removed. 29273683 9 99 theme CBG 1413:1415 arg1 binding 1402:1408 the steroid binding 1390:1408 the steroid binding of CBG 1390:1415 Thus, the presence and composition of an N-glycan in this conserved position both appear to influence the steroid binding of CBG. 25451932 0 0 theme membrane 70:77 arg1 targeting 79:87 cell membrane targeting 65:87 cell membrane targeting 65:87 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 1 1 theme essential 149:157 arg1 protease 140:147 a membrane-bound protease 123:147 a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure 123:223 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 1 1 theme essential 149:157 arg1 Corin 114:118 Corin 114:118 Corin 114:118 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 10 2 gly N-Glycosylation 1248:1262 arg2 Asn-1022 1267:1274 Asn-1022 1267:1274 Asn-1022 1267:1274 N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. 25451932 0 3 theme cell 65:68 arg1 targeting 79:87 cell membrane targeting 65:87 cell membrane targeting 65:87 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 6 4 theme juxtamembrane 848:860 arg1 domain 862:867 the juxtamembrane domain 844:867 the juxtamembrane domain 844:867 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 9 5 gly N-glycosylation 1186:1200 arg2 site 1202:1205 the N-glycosylation site 1182:1205 the N-glycosylation site 1182:1205 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 3 6 theme cell 363:366 arg1 expression 376:385 corin cell surface expression 357:385 corin cell surface expression 357:385 It has been shown that N-glycans are required for corin cell surface expression and zymogen activation. 25451932 9 7 theme protease 1214:1221 arg1 domain 1223:1228 the protease domain 1210:1228 the protease domain 1210:1228 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 9 8 theme N-glycosylation 1186:1200 arg1 site 1202:1205 the N-glycosylation site 1182:1205 the N-glycosylation site 1182:1205 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 6 9 from lysate 691:696 arg1 medium 714:719 conditioned medium 702:719 conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes 702:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 9 from lysate 691:696 arg1 analysis 656:663 Western analysis 648:663 Western analysis of corin proteins in cell lysate 648:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 7 10 from autocleavage 929:940 arg1 domain 960:965 the frizzled-1 domain 945:965 the frizzled-1 domain 945:965 Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. 25451932 3 11 theme surface 368:374 arg1 expression 376:385 corin cell surface expression 357:385 corin cell surface expression 357:385 It has been shown that N-glycans are required for corin cell surface expression and zymogen activation. 25451932 11 12 theme zymogen 1476:1482 arg1 activation 1484:1493 zymogen activation 1476:1493 zymogen activation 1476:1493 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 6 13 theme HL-1 755:758 arg1 cardiomyocytes 760:773 HL-1 cardiomyocytes 755:773 HL-1 cardiomyocytes 755:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 5 14 theme predicted 587:595 arg1 sites 613:617 the 19 predicted N-glycosylation sites 580:617 the 19 predicted N-glycosylation sites 580:617 In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. 25451932 11 15 theme membrane 1456:1463 arg1 targeting 1465:1473 the cell membrane targeting 1447:1473 the cell membrane targeting 1447:1473 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 6 16 theme conditioned 702:712 arg1 medium 714:719 conditioned medium 702:719 conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes 702:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 0 17 from sites 47:51 arg1 roles 9:13 Distinct roles 0:13 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 0:112 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 0 17 from sites 47:51 arg1 targeting 79:87 cell membrane targeting 65:87 cell membrane targeting 65:87 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 0 17 from sites 47:51 arg1 shedding 104:111 ectodomain shedding 93:111 ectodomain shedding 93:111 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 5 18 theme N-glycosylation 597:611 arg1 sites 613:617 the 19 predicted N-glycosylation sites 580:617 the 19 predicted N-glycosylation sites 580:617 In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. 25451932 2 19 theme extracellular 284:296 arg1 domains 298:304 its extracellular domains 280:304 its extracellular domains 280:304 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 10 20 theme different 1295:1303 arg1 sites 1305:1309 different sites 1295:1309 different sites 1295:1309 N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. 25451932 9 21 from location 1170:1177 arg1 domain 1223:1228 the protease domain 1210:1228 the protease domain 1210:1228 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 0 22 theme ectodomain 93:102 arg1 shedding 104:111 ectodomain shedding 93:111 ectodomain shedding 93:111 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 6 23 from shedding 832:839 arg1 domain 862:867 the juxtamembrane domain 844:867 the juxtamembrane domain 844:867 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 7 24 theme frizzled-1 949:958 arg1 domain 960:965 the frizzled-1 domain 945:965 the frizzled-1 domain 945:965 Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. 25451932 7 25 from Asn-231 900:906 arg1 N-glycosylation 881:895 N-glycosylation 881:895 N-glycosylation at Asn-231 881:906 Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. 25451932 1 26 theme natriuretic 174:184 arg1 peptides 186:193 natriuretic peptides 174:193 natriuretic peptides 174:193 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 3 27 theme zymogen 391:397 arg1 activation 399:408 zymogen activation 391:408 zymogen activation 391:408 It has been shown that N-glycans are required for corin cell surface expression and zymogen activation. 25451932 4 28 theme corin 486:490 arg1 biosynthesis 492:503 corin biosynthesis 486:503 corin biosynthesis 486:503 It remains unknown, however, how N-glycans at different sites may regulate corin biosynthesis and processing. 25451932 8 29 theme scavenger 1012:1020 arg1 domain 1031:1036 the scavenger receptor domain 1008:1036 the scavenger receptor domain 1008:1036 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 10 30 from Asn-1022 1267:1274 arg1 N-Glycosylation 1248:1262 N-Glycosylation 1248:1262 N-Glycosylation at Asn-1022 1248:1274 N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. 25451932 6 31 theme cell 686:689 arg1 lysate 691:696 cell lysate 686:696 cell lysate 686:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 0 32 theme Distinct 0:7 arg1 roles 9:13 Distinct roles 0:13 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 0:112 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 11 33 from sites 1403:1407 arg1 N-glycans 1380:1388 N-glycans 1380:1388 N-glycans at different sites 1380:1407 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 10 34 theme zymogen 1328:1334 arg1 activation 1336:1345 corin zymogen activation 1322:1345 corin zymogen activation 1322:1345 N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. 25451932 6 35 from Asn-80 809:814 arg1 N-glycosylation 790:804 N-glycosylation 790:804 N-glycosylation at Asn-80 790:814 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 0 36 theme N-glycosylation 18:32 arg1 roles 9:13 Distinct roles 0:13 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 0:112 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 4 37 from sites 467:471 arg1 N-glycans 444:452 N-glycans 444:452 N-glycans at different sites 444:471 It remains unknown, however, how N-glycans at different sites may regulate corin biosynthesis and processing. 25451932 6 38 theme proteins 674:681 arg1 medium 714:719 conditioned medium 702:719 conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes 702:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 38 theme proteins 674:681 arg1 analysis 656:663 Western analysis 648:663 Western analysis of corin proteins in cell lysate 648:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 8 39 from N-glycosylation 978:992 arg1 domain 1031:1036 the scavenger receptor domain 1008:1036 the scavenger receptor domain 1008:1036 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 2 40 gly N-glycosylation 255:269 arg2 sites 271:275 19 predicted N-glycosylation sites 242:275 19 predicted N-glycosylation sites 242:275 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 2 40 gly N-glycosylation 255:269 arg2 19 242:243 19 242:243 19 242:243 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 8 41 from Asn-1022 1045:1052 arg1 domain 1070:1075 the protease domain 1057:1075 the protease domain 1057:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 41 from Asn-1022 1045:1052 arg1 N-glycosylation 978:992 N-glycosylation 978:992 N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain 978:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 6 42 from proteins 674:681 arg1 lysate 691:696 cell lysate 686:696 cell lysate 686:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 43 theme corin 668:672 arg1 proteins 674:681 corin proteins 668:681 corin proteins in cell lysate 668:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 1 44 theme blood 210:214 arg1 pressure 216:223 blood pressure 210:223 blood pressure 210:223 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 0 45 theme different 37:45 arg1 corin 56:60 corin 56:60 corin 56:60 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 0 45 theme different 37:45 arg1 sites 47:51 different sites 37:51 different sites of corin in cell membrane targeting and ectodomain shedding 37:111 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 8 46 from Asn-697 997:1003 arg1 N-glycosylation 978:992 N-glycosylation 978:992 N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain 978:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 47 theme surface 1105:1111 arg1 targeting 1113:1121 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 11 48 theme cell 1451:1454 arg1 targeting 1465:1473 the cell membrane targeting 1447:1473 the cell membrane targeting 1447:1473 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 8 49 theme cell 1100:1103 arg1 targeting 1113:1121 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 7 50 gly N-glycosylation 881:895 arg1 Asn-231 900:906 Asn-231 900:906 Asn-231 900:906 Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. 25451932 7 50 gly N-glycosylation 881:895 arg2 Asn-231 900:906 Asn-231 900:906 Asn-231 900:906 Similarly, N-glycosylation at Asn-231 protected corin from autocleavage in the frizzled-1 domain. 25451932 5 51 gly N-glycosylation 597:611 arg2 sites 613:617 the 19 predicted N-glycosylation sites 580:617 the 19 predicted N-glycosylation sites 580:617 In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. 25451932 5 51 gly N-glycosylation 597:611 arg2 19 584:585 19 584:585 19 584:585 In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. 25451932 8 52 theme protease 1061:1068 arg1 domain 1070:1075 the protease domain 1057:1075 the protease domain 1057:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 3 53 theme corin 357:361 arg1 expression 376:385 corin cell surface expression 357:385 corin cell surface expression 357:385 It has been shown that N-glycans are required for corin cell surface expression and zymogen activation. 25451932 11 54 theme distinct 1418:1425 arg1 roles 1427:1431 distinct roles 1418:1431 distinct roles 1418:1431 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 9 55 theme site 1202:1205 arg1 location 1170:1177 the location 1166:1177 the location of the N-glycosylation site in the protease domain 1166:1228 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 9 55 theme site 1202:1205 arg1 critical 1238:1245 critical 1238:1245 critical 1238:1245 We also found that the location of the N-glycosylation site in the protease domain was not critical. 25451932 4 56 theme different 457:465 arg1 sites 467:471 different sites 457:471 different sites 457:471 It remains unknown, however, how N-glycans at different sites may regulate corin biosynthesis and processing. 25451932 6 57 from analysis 656:663 arg1 lysate 691:696 cell lysate 686:696 cell lysate 686:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 5 58 theme corin 548:552 arg1 mutants 554:560 corin mutants 548:560 corin mutants 548:560 In this study, we examined corin mutants, in which each of the 19 predicted N-glycosylation sites was mutated individually. 25451932 6 59 gly N-glycosylation 790:804 arg2 Asn-80 809:814 Asn-80 809:814 Asn-80 809:814 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 59 gly N-glycosylation 790:804 arg1 Asn-80 809:814 Asn-80 809:814 Asn-80 809:814 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 8 60 theme corin 1094:1098 arg1 targeting 1113:1121 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 2 61 theme Human 226:230 arg1 corin 232:236 Human corin 226:236 Human corin 226:236 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 6 62 from cells 745:749 arg1 medium 714:719 conditioned medium 702:719 conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes 702:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 62 from cells 745:749 arg1 analysis 656:663 Western analysis 648:663 Western analysis of corin proteins in cell lysate 648:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 8 63 theme receptor 1022:1029 arg1 domain 1031:1036 the scavenger receptor domain 1008:1036 the scavenger receptor domain 1008:1036 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 6 64 from cardiomyocytes 760:773 arg1 medium 714:719 conditioned medium 702:719 conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes 702:773 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 64 from cardiomyocytes 760:773 arg1 analysis 656:663 Western analysis 648:663 Western analysis of corin proteins in cell lysate 648:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 6 65 theme Western 648:654 arg1 analysis 656:663 Western analysis 648:663 Western analysis of corin proteins in cell lysate 648:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 8 66 theme zymogen 1127:1133 arg1 activation 1135:1144 zymogen activation 1127:1144 zymogen activation 1127:1144 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 6 67 theme HEK293 738:743 arg1 cells 745:749 transfected HEK293 cells 726:749 transfected HEK293 cells 726:749 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 10 68 theme corin 1322:1326 arg1 activation 1336:1345 corin zymogen activation 1322:1345 corin zymogen activation 1322:1345 N-Glycosylation at Asn-1022 may be switched to different sites to promote corin zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 zymogen 1127:1133 zymogen activation 1127:1144 zymogen activation 1127:1144 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 Asn-1022 1045:1052 Asn-1022 1045:1052 Asn-1022 in the protease domain 1045:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 corin 1094:1098 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg2 Asn-697 997:1003 Asn-697 997:1003 Asn-697 997:1003 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 cell 1100:1103 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 surface 1105:1111 corin cell surface targeting 1094:1121 corin cell surface targeting 1094:1121 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 domain 1031:1036 the scavenger receptor domain 1008:1036 the scavenger receptor domain 1008:1036 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg2 Asn-1022 1045:1052 Asn-1022 1045:1052 Asn-1022 in the protease domain 1045:1075 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 8 69 gly N-glycosylation 978:992 arg1 Asn-697 997:1003 Asn-697 997:1003 Asn-697 997:1003 Moreover, N-glycosylation at Asn-697 in the scavenger receptor domain and at Asn-1022 in the protease domain is important for corin cell surface targeting and zymogen activation. 25451932 11 70 theme different 1393:1401 arg1 sites 1403:1407 different sites 1393:1407 different sites 1393:1407 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 6 71 theme corin 826:830 arg1 shedding 832:839 corin shedding 826:839 corin shedding in the juxtamembrane domain 826:867 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 2 72 theme N-glycosylation 255:269 arg1 sites 271:275 19 predicted N-glycosylation sites 242:275 19 predicted N-glycosylation sites 242:275 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 6 73 theme transfected 726:736 arg1 cells 745:749 transfected HEK293 cells 726:749 transfected HEK293 cells 726:749 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 1 74 theme membrane-bound 125:138 arg1 protease 140:147 a membrane-bound protease 123:147 a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure 123:223 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 1 74 theme membrane-bound 125:138 arg1 Corin 114:118 Corin 114:118 Corin 114:118 Corin is a membrane-bound protease essential for activating natriuretic peptides and regulating blood pressure. 25451932 11 75 theme ectodomain 1500:1509 arg1 shedding 1511:1518 ectodomain shedding 1500:1518 ectodomain shedding 1500:1518 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 2 76 contain has 238:240 arg2 sites 271:275 19 predicted N-glycosylation sites 242:275 19 predicted N-glycosylation sites 242:275 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 2 76 contain has 238:240 arg1 corin 232:236 Human corin 226:236 Human corin 226:236 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 0 77 theme corin 56:60 arg1 corin 56:60 corin 56:60 corin 56:60 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 0 77 theme corin 56:60 arg1 sites 47:51 different sites 37:51 different sites of corin in cell membrane targeting and ectodomain shedding 37:111 Distinct roles of N-glycosylation at different sites of corin in cell membrane targeting and ectodomain shedding. 25451932 2 78 theme predicted 245:253 arg1 sites 271:275 19 predicted N-glycosylation sites 242:275 19 predicted N-glycosylation sites 242:275 Human corin has 19 predicted N-glycosylation sites in its extracellular domains. 25451932 6 79 from medium 714:719 arg1 lysate 691:696 cell lysate 686:696 cell lysate 686:696 By Western analysis of corin proteins in cell lysate and conditioned medium from transfected HEK293 cells and HL-1 cardiomyocytes, we found that N-glycosylation at Asn-80 inhibited corin shedding in the juxtamembrane domain. 25451932 11 80 theme corin 1523:1527 arg1 targeting 1465:1473 the cell membrane targeting 1447:1473 the cell membrane targeting 1447:1473 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 11 80 theme corin 1523:1527 arg1 activation 1484:1493 zymogen activation 1476:1493 zymogen activation 1476:1493 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 25451932 11 80 theme corin 1523:1527 arg1 shedding 1511:1518 ectodomain shedding 1500:1518 ectodomain shedding 1500:1518 Together, our results show that N-glycans at different sites may play distinct roles in regulating the cell membrane targeting, zymogen activation, and ectodomain shedding of corin. 26598643 7 0 attach derived 1348:1354 arg2 composition 1331:1341 glycan composition 1324:1341 glycan composition 1324:1341 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 7 0 attach derived 1348:1354 arg2 identity 1311:1318 Peptide identity 1303:1318 Peptide identity 1303:1318 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 7 0 attach derived 1348:1354 arg1 spectra 1412:1418 low-energy collision-induced dissociation fragment spectra 1361:1418 low-energy collision-induced dissociation fragment spectra acquired in multistage mode 1361:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 10 1 theme acute-phase 1727:1737 arg1 majority 1712:1719 the majority 1708:1719 the majority 1708:1719 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 10 1 theme acute-phase 1727:1737 arg1 proteins 1739:1746 acute-phase proteins 1727:1746 acute-phase proteins 1727:1746 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 15 2 theme human 2204:2208 arg1 O-glycoproteome 2223:2237 the human blood plasma O-glycoproteome 2200:2237 the human blood plasma O-glycoproteome 2200:2237 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 5 3 from O-glycans 1059:1067 arg1 proteins 1091:1098 human blood plasma proteins 1072:1098 human blood plasma proteins 1072:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 16 4 theme ProteomeXchange 2364:2378 arg1 PXD003270 2380:2388 ProteomeXchange PXD003270 2364:2388 ProteomeXchange PXD003270 2364:2388 The data were deposited to ProteomeXchange PXD003270. 26598643 5 5 theme fractionation 847:859 arg1 step 861:864 fractionation step 847:864 fractionation step 847:864 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 6 from majority 1047:1054 arg1 proteins 1091:1098 human blood plasma proteins 1072:1098 human blood plasma proteins 1072:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 14 7 theme identified 1978:1987 arg1 compositions 1998:2009 The identified O-glycan compositions 1974:2009 The identified O-glycan compositions 1974:2009 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 6 8 theme trap 1249:1252 arg1 spectrometer 1259:1270 an ion trap mass spectrometer 1242:1270 an ion trap mass spectrometer operated in positive-ion mode 1242:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 5 9 theme donors 690:695 arg1 plasma 672:677 pooled human blood plasma 653:677 pooled human blood plasma of healthy donors 653:695 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 2 10 theme O-glycosylated 323:336 arg1 proteins 338:345 O-glycosylated proteins 323:345 O-glycosylated proteins 323:345 The analysis, though, is quite challenging and time consuming, in particular for O-glycosylated proteins. 26598643 8 11 dep fragmented 1506:1515 arg1 pinpoint 1452:1459 pinpoint 1452:1459 To pinpoint the O-glycosylation sites 1449:1485 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 11 12 gly O-glycosylation 1775:1789 arg2 sites 1791:1795 11 novel O-glycosylation sites 1766:1795 11 novel O-glycosylation sites 1766:1795 Strikingly, also 11 novel O-glycosylation sites and regions were identified. 26598643 11 12 gly O-glycosylation 1775:1789 arg2 11 1766:1767 11 1766:1767 11 1766:1767 Strikingly, also 11 novel O-glycosylation sites and regions were identified. 26598643 5 13 theme interaction 882:892 arg1 chromatography 901:914 hydrophilic interaction liquid chromatography 870:914 hydrophilic interaction liquid chromatography 870:914 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 14 14 theme mono- 2039:2043 arg1 T-antigen 2091:2099 T-antigen 2091:2099 T-antigen 2091:2099 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 14 theme mono- 2039:2043 arg1 O-glycans 2080:2088 core-1 mucin-type O-glycans 2062:2088 mono- and disialylated core-1 mucin-type O-glycans (T-antigen) 2039:2100 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 15 theme disialylated 2049:2060 arg1 T-antigen 2091:2099 T-antigen 2091:2099 T-antigen 2091:2099 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 15 theme disialylated 2049:2060 arg1 O-glycans 2080:2088 core-1 mucin-type O-glycans 2062:2088 mono- and disialylated core-1 mucin-type O-glycans (T-antigen) 2039:2100 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 13 16 theme Proteinase 1907:1916 arg1 K 1918:1918 Proteinase K 1907:1918 Proteinase K 1907:1918 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 5 17 theme broad-specific 734:747 arg1 enzyme 749:754 a broad-specific enzyme 732:754 a broad-specific enzyme (Proteinase K) 732:769 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 17 theme broad-specific 734:747 arg1 K 768:768 Proteinase K 757:768 Proteinase K 757:768 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 18 theme short 980:984 arg1 core-1 997:1002 short mucin-type core-1 980:1002 short mucin-type core-1 980:1002 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 19 gly O-glycopeptides 955:969 arg2 O-glycopeptides 955:969 intact O-glycopeptides 948:969 intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins 948:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 20 contain carrying 971:978 arg2 core-1 997:1002 short mucin-type core-1 980:1002 short mucin-type core-1 980:1002 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 20 contain carrying 971:978 arg1 O-glycopeptides 955:969 intact O-glycopeptides 948:969 intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins 948:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 20 contain carrying 971:978 arg2 O-glycans 1011:1019 -2 O-glycans 1008:1019 -2 O-glycans 1008:1019 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 13 21 from context 1965:1971 arg1 beneficial 1946:1955 beneficial 1946:1955 beneficial 1946:1955 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 13 21 from context 1965:1971 arg1 use 1900:1902 the use 1896:1902 the use of Proteinase K 1896:1918 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 1 22 theme glycoproteins 166:178 arg1 relationships 149:161 structure-function relationships 130:161 structure-function relationships of glycoproteins, e.g. 130:184 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 4 23 gly O-glycosylation 579:593 arg1 glycoproteins 626:638 human blood plasma glycoproteins 607:638 human blood plasma glycoproteins 607:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 14 24 theme O-glycan 1989:1996 arg1 compositions 1998:2009 The identified O-glycan compositions 1974:2009 The identified O-glycan compositions 1974:2009 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 3 25 theme clinical 378:385 arg1 importance 409:418 their clinical and biopharmaceutical importance 372:418 their clinical and biopharmaceutical importance 372:418 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 7 26 theme Peptide 1303:1309 arg1 identity 1311:1318 Peptide identity 1303:1318 Peptide identity 1303:1318 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 5 27 theme human 660:664 arg1 plasma 672:677 pooled human blood plasma 653:677 pooled human blood plasma of healthy donors 653:695 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 1 28 theme Site-specific 71:83 arg1 analysis 99:106 Site-specific glycosylation analysis 71:106 Site-specific glycosylation analysis 71:106 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 12 29 gly O-glycosylation 1838:1852 arg2 23 1835:1836 23 1835:1836 23 1835:1836 In total 23 O-glycosylation sites could be pinpointed. 26598643 12 29 gly O-glycosylation 1838:1852 arg2 sites 1854:1858 23 O-glycosylation sites 1835:1858 23 O-glycosylation sites 1835:1858 In total 23 O-glycosylation sites could be pinpointed. 26598643 5 30 theme precipitation 786:798 arg1 step 800:803 a precipitation step 784:803 a precipitation step 784:803 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 4 31 gly glycoproteins 626:638 arg1 glycoproteins 626:638 human blood plasma glycoproteins 607:638 human blood plasma glycoproteins 607:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 5 32 theme vast 1042:1045 arg1 majority 1047:1054 the vast majority 1038:1054 the vast majority of O-glycans on human blood plasma proteins 1038:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 0 33 theme Site-specific 0:12 arg1 Analysis 30:37 Site-specific O-Glycosylation Analysis 0:37 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 0:69 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 8 34 gly O-glycosylation 1465:1479 arg2 sites 1481:1485 the O-glycosylation sites 1461:1485 the O-glycosylation sites 1461:1485 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 3 35 theme biopharmaceutical 391:407 arg1 importance 409:418 their clinical and biopharmaceutical importance 372:418 their clinical and biopharmaceutical importance 372:418 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 6 36 gly O-glycopeptide 1110:1123 arg2 O-glycopeptide 1110:1123 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 8 37 theme transfer 1532:1539 arg1 dissociation 1541:1552 electron transfer dissociation 1523:1552 electron transfer dissociation 1523:1552 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 15 38 theme structure-function 2303:2320 arg1 relationships 2322:2334 structure-function relationships 2303:2334 structure-function relationships 2303:2334 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 8 39 gly glycopeptides 1487:1499 arg2 glycopeptides 1487:1499 glycopeptides 1487:1499 glycopeptides 1487:1499 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 15 40 theme O-glycoproteome 2223:2237 arg1 population 2186:2195 the major population 2176:2195 the major population of the human blood plasma O-glycoproteome 2176:2237 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 6 41 theme O-glycopeptide 1110:1123 arg1 fractions 1125:1133 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 3 42 theme plasma 438:443 arg1 glycoproteins 445:457 many human blood plasma glycoproteins 421:457 many human blood plasma glycoproteins 421:457 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 4 43 theme blood 613:617 arg1 glycoproteins 626:638 human blood plasma glycoproteins 607:638 human blood plasma glycoproteins 607:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 0 44 theme Human 42:46 arg1 Proteins 61:68 Human Blood Plasma Proteins 42:68 Human Blood Plasma Proteins 42:68 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 5 45 theme blood 1078:1082 arg1 proteins 1091:1098 human blood plasma proteins 1072:1098 human blood plasma proteins 1072:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 3 46 theme human 426:430 arg1 glycoproteins 445:457 many human blood plasma glycoproteins 421:457 many human blood plasma glycoproteins 421:457 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 15 47 theme major 2180:2184 arg1 population 2186:2195 the major population 2176:2195 the major population of the human blood plasma O-glycoproteome 2176:2237 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 0 48 theme Plasma 54:59 arg1 Proteins 61:68 Human Blood Plasma Proteins 42:68 Human Blood Plasma Proteins 42:68 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 4 49 theme O-glycosylation 579:593 arg1 analysis 595:602 the site-specific O-glycosylation analysis 561:602 the site-specific O-glycosylation analysis of human blood plasma glycoproteins 561:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 6 50 theme liquid 1202:1207 arg1 chromatography 1209:1222 reversed-phase liquid chromatography 1187:1222 reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode 1187:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 5 51 dep end 649:651 arg1 digested 717:724 digested 717:724 digested using a broad-specific enzyme (Proteinase K) 717:769 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 2 52 gly O-glycosylated 323:336 arg1 proteins 338:345 O-glycosylated proteins 323:345 O-glycosylated proteins 323:345 The analysis, though, is quite challenging and time consuming, in particular for O-glycosylated proteins. 26598643 0 53 theme Proteins 61:68 arg1 Analysis 30:37 Site-specific O-Glycosylation Analysis 0:37 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 0:69 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 7 54 theme low-energy 1361:1370 arg1 spectra 1412:1418 low-energy collision-induced dissociation fragment spectra 1361:1418 low-energy collision-induced dissociation fragment spectra acquired in multistage mode 1361:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 11 55 theme novel 1769:1773 arg1 sites 1791:1795 11 novel O-glycosylation sites 1766:1795 11 novel O-glycosylation sites 1766:1795 Strikingly, also 11 novel O-glycosylation sites and regions were identified. 26598643 6 56 theme spectrometric 1158:1170 arg1 analysis 1172:1179 mass spectrometric analysis 1153:1179 mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode 1153:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 4 57 theme glycoproteins 626:638 arg1 analysis 595:602 the site-specific O-glycosylation analysis 561:602 the site-specific O-glycosylation analysis of human blood plasma glycoproteins 561:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 1 58 theme progression 229:239 arg1 context 193:199 the context 189:199 the context of antigenicity and disease progression 189:239 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 9 59 theme database 1581:1588 arg1 searches 1590:1597 database searches 1581:1597 database searches 1581:1597 Spectra were annotated by database searches as well as manually. 26598643 6 60 theme mass 1254:1257 arg1 spectrometer 1259:1270 an ion trap mass spectrometer 1242:1270 an ion trap mass spectrometer operated in positive-ion mode 1242:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 13 61 theme K 1918:1918 arg1 use 1900:1902 the use 1896:1902 the use of Proteinase K 1896:1918 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 13 61 theme K 1918:1918 arg1 beneficial 1946:1955 beneficial 1946:1955 beneficial 1946:1955 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 5 62 theme healthy 682:688 arg1 donors 690:695 healthy donors 682:695 healthy donors 682:695 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 1 63 theme antigenicity 204:215 arg1 context 193:199 the context 189:199 the context of antigenicity and disease progression 189:239 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 6 64 theme ion 1245:1247 arg1 spectrometer 1259:1270 an ion trap mass spectrometer 1242:1270 an ion trap mass spectrometer operated in positive-ion mode 1242:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 5 65 theme Proteinase 757:766 arg1 enzyme 749:754 a broad-specific enzyme 732:754 a broad-specific enzyme (Proteinase K) 732:769 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 65 theme Proteinase 757:766 arg1 K 768:768 Proteinase K 757:768 Proteinase K 757:768 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 66 from proteins 1091:1098 arg1 majority 1047:1054 the vast majority 1038:1054 the vast majority of O-glycans on human blood plasma proteins 1038:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 1 67 theme disease 221:227 arg1 progression 229:239 disease progression 221:239 disease progression 221:239 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 10 68 gly O-glycosylation 1632:1646 arg2 31 1629:1630 31 1629:1630 31 1629:1630 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 10 68 gly O-glycosylation 1632:1646 arg2 sites 1648:1652 31 O-glycosylation sites 1629:1652 31 O-glycosylation sites 1629:1652 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 1 69 gly glycoproteins 166:178 arg1 glycoproteins 166:178 glycoproteins 166:178 glycoproteins 166:178 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 5 70 theme liquid 894:899 arg1 chromatography 901:914 hydrophilic interaction liquid chromatography 870:914 hydrophilic interaction liquid chromatography 870:914 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 10 71 dep identified 1696:1705 arg1 majority 1712:1719 the majority 1708:1719 the majority 1708:1719 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 10 71 dep identified 1696:1705 arg1 proteins 1739:1746 acute-phase proteins 1727:1746 acute-phase proteins 1727:1746 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 13 72 from beneficial 1946:1955 arg1 context 1965:1971 this context 1960:1971 this context 1960:1971 Interestingly, the use of Proteinase K proved to be particularly beneficial in this context. 26598643 1 73 theme structure-function 130:147 arg1 relationships 149:161 structure-function relationships 130:161 structure-function relationships of glycoproteins, e.g. 130:184 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 5 74 theme mucin-type 986:995 arg1 core-1 997:1002 short mucin-type core-1 980:1002 short mucin-type core-1 980:1002 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 14 75 theme mucin-type 2069:2078 arg1 T-antigen 2091:2099 T-antigen 2091:2099 T-antigen 2091:2099 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 75 theme mucin-type 2069:2078 arg1 O-glycans 2080:2088 core-1 mucin-type O-glycans 2062:2088 mono- and disialylated core-1 mucin-type O-glycans (T-antigen) 2039:2100 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 7 76 theme glycan 1324:1329 arg1 composition 1331:1341 glycan composition 1324:1341 glycan composition 1324:1341 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 5 77 theme hydrophilic 870:880 arg1 chromatography 901:914 hydrophilic interaction liquid chromatography 870:914 hydrophilic interaction liquid chromatography 870:914 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 6 78 theme positive-ion 1284:1295 arg1 mode 1297:1300 positive-ion mode 1284:1300 positive-ion mode 1284:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 3 79 gly glycoproteins 445:457 arg1 glycoproteins 445:457 many human blood plasma glycoproteins 421:457 many human blood plasma glycoproteins 421:457 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 5 80 theme intact 948:953 arg1 O-glycopeptides 955:969 intact O-glycopeptides 948:969 intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins 948:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 12 81 theme O-glycosylation 1838:1852 arg1 sites 1854:1858 23 O-glycosylation sites 1835:1858 23 O-glycosylation sites 1835:1858 In total 23 O-glycosylation sites could be pinpointed. 26598643 5 82 theme pooled 653:658 arg1 plasma 672:677 pooled human blood plasma 653:677 pooled human blood plasma of healthy donors 653:695 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 15 83 theme new 2263:2265 arg1 insights 2267:2274 new insights 2263:2274 new insights 2263:2274 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 8 84 theme O-glycosylation 1465:1479 arg1 sites 1481:1485 the O-glycosylation sites 1461:1485 the O-glycosylation sites 1461:1485 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 7 85 theme dissociation 1390:1401 arg1 spectra 1412:1418 low-energy collision-induced dissociation fragment spectra 1361:1418 low-energy collision-induced dissociation fragment spectra acquired in multistage mode 1361:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 5 86 theme blood 666:670 arg1 plasma 672:677 pooled human blood plasma 653:677 pooled human blood plasma of healthy donors 653:695 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 1 87 theme glycosylation 85:97 arg1 analysis 99:106 Site-specific glycosylation analysis 71:106 Site-specific glycosylation analysis 71:106 Site-specific glycosylation analysis is key to investigate structure-function relationships of glycoproteins, e.g. in the context of antigenicity and disease progression. 26598643 6 88 theme mass 1153:1156 arg1 analysis 1172:1179 mass spectrometric analysis 1153:1179 mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode 1153:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 0 89 theme O-Glycosylation 14:28 arg1 Analysis 30:37 Site-specific O-Glycosylation Analysis 0:37 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 0:69 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 3 90 theme many 421:424 arg1 glycoproteins 445:457 many human blood plasma glycoproteins 421:457 many human blood plasma glycoproteins 421:457 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 4 91 theme human 607:611 arg1 glycoproteins 626:638 human blood plasma glycoproteins 607:638 human blood plasma glycoproteins 607:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 15 92 theme developed 2107:2115 arg1 workflow 2117:2124 The developed workflow 2103:2124 The developed workflow 2103:2124 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 14 93 gly disialylated 2049:2060 arg1 T-antigen 2091:2099 T-antigen 2091:2099 T-antigen 2091:2099 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 93 gly disialylated 2049:2060 arg1 O-glycans 2080:2088 core-1 mucin-type O-glycans 2062:2088 mono- and disialylated core-1 mucin-type O-glycans (T-antigen) 2039:2100 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 15 94 theme plasma 2216:2221 arg1 O-glycoproteome 2223:2237 the human blood plasma O-glycoproteome 2200:2237 the human blood plasma O-glycoproteome 2200:2237 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 7 95 theme multistage 1432:1441 arg1 mode 1443:1446 multistage mode 1432:1446 multistage mode 1432:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 4 96 theme plasma 619:624 arg1 glycoproteins 626:638 human blood plasma glycoproteins 607:638 human blood plasma glycoproteins 607:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 15 97 theme blood 2210:2214 arg1 O-glycoproteome 2223:2237 the human blood plasma O-glycoproteome 2200:2237 the human blood plasma O-glycoproteome 2200:2237 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 8 98 theme electron 1523:1530 arg1 dissociation 1541:1552 electron transfer dissociation 1523:1552 electron transfer dissociation 1523:1552 To pinpoint the O-glycosylation sites glycopeptides were fragmented using electron transfer dissociation. 26598643 0 99 theme Blood 48:52 arg1 Proteins 61:68 Human Blood Plasma Proteins 42:68 Human Blood Plasma Proteins 42:68 Site-specific O-Glycosylation Analysis of Human Blood Plasma Proteins. 26598643 14 100 theme core-1 2062:2067 arg1 T-antigen 2091:2099 T-antigen 2091:2099 T-antigen 2091:2099 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 14 100 theme core-1 2062:2067 arg1 O-glycans 2080:2088 core-1 mucin-type O-glycans 2062:2088 mono- and disialylated core-1 mucin-type O-glycans (T-antigen) 2039:2100 The identified O-glycan compositions most probably correspond to mono- and disialylated core-1 mucin-type O-glycans (T-antigen). 26598643 5 101 theme human 1072:1076 arg1 proteins 1091:1098 human blood plasma proteins 1072:1098 human blood plasma proteins 1072:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 6 102 theme Enriched 1101:1108 arg1 fractions 1125:1133 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions 1101:1133 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 4 103 theme site-specific 565:577 arg1 analysis 595:602 the site-specific O-glycosylation analysis 561:602 the site-specific O-glycosylation analysis of human blood plasma glycoproteins 561:638 Here, we report on the site-specific O-glycosylation analysis of human blood plasma glycoproteins. 26598643 10 104 theme O-glycosylation 1632:1646 arg1 sites 1648:1652 31 O-glycosylation sites 1629:1652 31 O-glycosylation sites 1629:1652 Overall, 31 O-glycosylation sites and regions belonging to 22 proteins were identified, the majority being acute-phase proteins. 26598643 5 105 theme glycopeptide 819:830 arg1 enrichment 832:841 a glycopeptide enrichment 817:841 a glycopeptide enrichment 817:841 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 5 106 theme plasma 1084:1089 arg1 proteins 1091:1098 human blood plasma proteins 1072:1098 human blood plasma proteins 1072:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 7 107 theme fragment 1403:1410 arg1 spectra 1412:1418 low-energy collision-induced dissociation fragment spectra 1361:1418 low-energy collision-induced dissociation fragment spectra acquired in multistage mode 1361:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 3 108 theme blood 432:436 arg1 glycoproteins 445:457 many human blood plasma glycoproteins 421:457 many human blood plasma glycoproteins 421:457 In consequence, despite their clinical and biopharmaceutical importance, many human blood plasma glycoproteins have not been characterized comprehensively with respect to their O-glycosylation. 26598643 15 109 theme population 2186:2195 arg1 identification 2137:2150 identification 2137:2150 identification 2137:2150 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 15 109 theme population 2186:2195 arg1 characterization 2156:2171 characterization 2156:2171 characterization 2156:2171 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 7 110 theme collision-induced 1372:1388 arg1 spectra 1412:1418 low-energy collision-induced dissociation fragment spectra 1361:1418 low-energy collision-induced dissociation fragment spectra acquired in multistage mode 1361:1446 Peptide identity and glycan composition were derived from low-energy collision-induced dissociation fragment spectra acquired in multistage mode. 26598643 15 111 dep identification 2137:2150 arg1 the 2133:2135 the 2133:2135 the 2133:2135 The developed workflow allows the identification and characterization of the major population of the human blood plasma O-glycoproteome and our results provide new insights, which can help to unravel structure-function relationships. 26598643 11 112 theme O-glycosylation 1775:1789 arg1 sites 1791:1795 11 novel O-glycosylation sites 1766:1795 11 novel O-glycosylation sites 1766:1795 Strikingly, also 11 novel O-glycosylation sites and regions were identified. 26598643 5 113 theme O-glycans 1059:1067 arg1 majority 1047:1054 the vast majority 1038:1054 the vast majority of O-glycans on human blood plasma proteins 1038:1098 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 26598643 6 114 theme reversed-phase 1187:1200 arg1 chromatography 1209:1222 reversed-phase liquid chromatography 1187:1222 reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode 1187:1300 Enriched O-glycopeptide fractions were subjected to mass spectrometric analysis using reversed-phase liquid chromatography coupled online to an ion trap mass spectrometer operated in positive-ion mode. 26598643 5 115 gly glycopeptide 819:830 arg2 glycopeptide 819:830 a glycopeptide enrichment 817:841 a glycopeptide enrichment 817:841 To this end pooled human blood plasma of healthy donors was proteolytically digested using a broad-specific enzyme (Proteinase K), followed by a precipitation step, as well as a glycopeptide enrichment and fractionation step via hydrophilic interaction liquid chromatography, the latter being optimized for intact O-glycopeptides carrying short mucin-type core-1 and -2 O-glycans, which represent the vast majority of O-glycans on human blood plasma proteins. 25533529 7 0 theme mice 1475:1478 arg1 model 1480:1484 the mice model 1471:1484 the mice model 1471:1484 In the mice model, the defibrinogenating effect of ATB-wt was stronger and lasting-longer than those of all the mutants. 25533529 4 1 theme enzyme 838:843 arg1 folding 845:851 the enzyme folding 834:851 the enzyme folding 834:851 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 1 2 theme Deinagkistrodon 166:180 arg1 venom 189:193 Deinagkistrodon acutus venom 166:193 Deinagkistrodon acutus venom 166:193 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 3 3 gly -deglycosylated 600:614 arg1 mutants 616:622 various Asn(229)-deglycosylated mutants 584:622 various Asn(229)-deglycosylated mutants 584:622 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 5 4 theme active 1108:1113 arg1 sites 1115:1119 the active sites 1104:1119 the active sites 1104:1119 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 1 5 theme acutus 182:187 arg1 venom 189:193 Deinagkistrodon acutus venom 166:193 Deinagkistrodon acutus venom 166:193 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 3 6 from Asn 528:530 arg1 mutations 494:502 mutations 494:502 mutations at Asn(77), Asn(81) and Asn(100) 494:535 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 6 7 theme other 1284:1288 arg1 mutants 1290:1296 other mutants 1284:1296 other mutants that cleaved only the α-subunit 1284:1328 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 0 8 theme recombinants 104:115 arg1 activities 81:90 folding and defibrinogenating activities 51:90 folding and defibrinogenating activities of acutobin recombinants from HEK293T 51:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 0 9 from Effects 0:6 arg1 activities 81:90 folding and defibrinogenating activities 51:90 folding and defibrinogenating activities of acutobin recombinants from HEK293T 51:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 3 10 from Asn 516:518 arg1 mutations 494:502 mutations 494:502 mutations at Asn(77), Asn(81) and Asn(100) 494:535 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 2 11 theme each 314:317 arg1 roles 305:309 the functional roles 290:309 the functional roles of each of the N-glycan 290:333 Here, we explore the functional roles of each of the N-glycan by site-directed mutagenesis. 25533529 5 12 theme similar 974:980 arg1 stabilities 990:1000 similar thermal stabilities 974:1000 similar thermal stabilities as ATB-wt 974:1010 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 0 13 theme acutobin 95:102 arg1 recombinants 104:115 acutobin recombinants 95:115 acutobin recombinants from HEK293T 95:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 5 14 theme gross 1068:1072 arg1 conformation 1074:1085 gross conformation 1068:1085 gross conformation 1068:1085 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 4 15 theme various 723:729 arg1 enzymes 751:757 various venom thrombin-like enzymes 723:757 various venom thrombin-like enzymes 723:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 8 16 theme acutobin 1796:1803 arg1 specificity 1781:1791 the fibrinogen-chain specificity 1760:1791 the fibrinogen-chain specificity of acutobin 1760:1803 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 4 17 theme homology 656:663 arg1 modeling 665:672 homology modeling 656:672 homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes 656:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 4 18 theme acutobin 677:684 arg1 modeling 665:672 homology modeling 656:672 homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes 656:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 4 19 from site 826:829 arg1 importance 764:773 the importance 760:773 the importance of a hydrophilic environment at each glycosylation site to the enzyme folding 760:851 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 4 20 theme glycosylation 812:824 arg1 site 826:829 each glycosylation site 807:829 each glycosylation site 807:829 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 5 21 theme chromogenic 948:958 arg1 substrate 960:968 the chromogenic substrate 944:968 the chromogenic substrate 944:968 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 1 22 gly N-glycosylation 210:224 arg2 sites 226:230 four N-glycosylation sites 205:230 four N-glycosylation sites with disialylated complex-typed glycans 205:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 1 22 gly N-glycosylation 210:224 arg2 four 205:208 four 205:208 four 205:208 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 3 23 from Asn 507:509 arg1 mutations 494:502 mutations 494:502 mutations at Asn(77), Asn(81) and Asn(100) 494:535 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 2 24 theme functional 294:303 arg1 roles 305:309 the functional roles 290:309 the functional roles of each of the N-glycan 290:333 Here, we explore the functional roles of each of the N-glycan by site-directed mutagenesis. 25533529 3 25 theme -deglycosylated 600:614 arg1 mutants 616:622 various Asn(229)-deglycosylated mutants 584:622 various Asn(229)-deglycosylated mutants 584:622 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 1 26 gly disialylated 237:248 arg1 glycans 264:270 disialylated complex-typed glycans 237:270 disialylated complex-typed glycans 237:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 6 27 theme fibrinogen-peptide 1379:1396 arg1 A 1398:1398 fibrinogen-peptide A 1379:1398 fibrinogen-peptide A 1379:1398 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 4 28 theme sequence 699:706 arg1 alignment 708:716 multiple sequence alignment 690:716 multiple sequence alignment 690:716 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 6 29 dep able 1363:1366 arg1 coagulated 1409:1418 coagulated 1409:1418 thus coagulated human plasma slower than the other mutants did 1404:1465 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 6 29 dep able 1363:1366 arg1 release 1371:1377 release 1371:1377 to release fibrinogen-peptide A 1368:1398 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 6 30 theme slower 1433:1438 arg1 plasma 1426:1431 human plasma 1420:1431 human plasma slower than the other mutants did 1420:1465 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 6 31 theme in vitro 1258:1265 arg1 subunits 1220:1227 all human fibrinogen subunits 1199:1227 all human fibrinogen subunits faster but less specifically in vitro 1199:1265 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 6 32 theme other 1449:1453 arg1 mutants 1455:1461 the other mutants 1445:1461 the other mutants did 1445:1465 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 4 33 theme multiple 690:697 arg1 alignment 708:716 multiple sequence alignment 690:716 multiple sequence alignment 690:716 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 4 34 theme venom 731:735 arg1 enzymes 751:757 various venom thrombin-like enzymes 723:757 various venom thrombin-like enzymes 723:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 1 35 contain contains 196:203 arg2 sites 226:230 four N-glycosylation sites 205:230 four N-glycosylation sites with disialylated complex-typed glycans 205:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 1 35 contain contains 196:203 arg1 Acutobin 131:138 Acutobin 131:138 Acutobin 131:138 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 1 35 contain contains 196:203 arg1 α-fibrinogenase 145:159 the α-fibrinogenase 141:159 the α-fibrinogenase from Deinagkistrodon acutus venom 141:193 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 0 36 theme N-glycosylation 18:32 arg1 site 34:37 single N-glycosylation site knockout 11:46 single N-glycosylation site knockout 11:46 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 5 37 theme thermal 982:988 arg1 stabilities 990:1000 similar thermal stabilities 974:1000 similar thermal stabilities as ATB-wt 974:1010 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 8 38 theme ATB-wt 1672:1677 arg1 pharmacokinetics 1639:1654 the pharmacokinetics 1635:1654 the pharmacokinetics of acutobin and ATB-wt in vivo 1635:1685 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 4 39 theme environment 792:802 arg1 importance 764:773 the importance 760:773 the importance of a hydrophilic environment at each glycosylation site to the enzyme folding 760:851 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 5 40 theme catalytic 919:927 arg1 activities 929:938 similar catalytic activities 911:938 similar catalytic activities for the chromogenic substrate 911:968 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 6 41 theme faster 1229:1234 arg1 subunits 1220:1227 all human fibrinogen subunits 1199:1227 all human fibrinogen subunits faster but less specifically in vitro 1199:1265 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 1 42 theme N-glycosylation 210:224 arg1 sites 226:230 four N-glycosylation sites 205:230 four N-glycosylation sites with disialylated complex-typed glycans 205:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 0 43 theme single 11:16 arg1 site 34:37 single N-glycosylation site knockout 11:46 single N-glycosylation site knockout 11:46 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 4 44 with modeling 665:672 arg1 enzymes 751:757 various venom thrombin-like enzymes 723:757 various venom thrombin-like enzymes 723:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 5 45 theme similar 911:917 arg1 activities 929:938 similar catalytic activities 911:938 similar catalytic activities for the chromogenic substrate 911:968 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 7 46 theme ATB-wt 1519:1524 arg1 stronger 1530:1537 stronger 1530:1537 stronger 1530:1537 In the mice model, the defibrinogenating effect of ATB-wt was stronger and lasting-longer than those of all the mutants. 25533529 7 46 theme ATB-wt 1519:1524 arg1 effect 1509:1514 the defibrinogenating effect 1487:1514 the defibrinogenating effect of ATB-wt 1487:1524 In the mice model, the defibrinogenating effect of ATB-wt was stronger and lasting-longer than those of all the mutants. 25533529 4 47 theme alignment 708:716 arg1 modeling 665:672 homology modeling 656:672 homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes 656:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 8 48 theme Asn 1724:1726 arg1 -glycan 1732:1738 the Asn(229)-glycan 1720:1738 the Asn(229)-glycan 1720:1738 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 4 49 gly glycosylation 812:824 arg2 site 826:829 each glycosylation site 807:829 each glycosylation site 807:829 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 0 50 theme knockout 39:46 arg1 site 34:37 single N-glycosylation site knockout 11:46 single N-glycosylation site knockout 11:46 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 8 51 from positions 1828:1836 arg1 N-glycans 1815:1823 the N-glycans 1811:1823 the N-glycans at positions 77, 81 and 100 1811:1851 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 8 51 from positions 1828:1836 arg1 crucial 1857:1863 crucial 1857:1863 crucial 1857:1863 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 0 52 from activities 81:90 arg1 HEK293T 122:128 HEK293T 122:128 HEK293T 122:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 4 53 theme hydrophilic 780:790 arg1 environment 792:802 a hydrophilic environment 778:802 a hydrophilic environment 778:802 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 0 54 theme site 34:37 arg1 Effects 0:6 Effects 0:6 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 0:129 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 1 55 with sites 226:230 arg1 glycans 264:270 disialylated complex-typed glycans 237:270 disialylated complex-typed glycans 237:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 3 56 theme S79A 568:571 arg1 mutant 573:578 the S79A mutant 564:578 the S79A mutant 564:578 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 1 57 theme disialylated 237:248 arg1 glycans 264:270 disialylated complex-typed glycans 237:270 disialylated complex-typed glycans 237:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 0 58 theme folding 51:57 arg1 activities 81:90 folding and defibrinogenating activities 51:90 folding and defibrinogenating activities of acutobin recombinants from HEK293T 51:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 8 59 dep acutobin 1659:1666 arg1 in vivo 1679:1685 in vivo 1679:1685 in vivo 1679:1685 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 6 60 theme fibrinogen 1209:1218 arg1 subunits 1220:1227 all human fibrinogen subunits 1199:1227 all human fibrinogen subunits faster but less specifically in vitro 1199:1265 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 3 61 theme recombinant 426:436 arg1 acutobin 438:445 recombinant acutobin 426:445 recombinant acutobin 426:445 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 0 62 from HEK293T 122:128 arg1 activities 81:90 folding and defibrinogenating activities 51:90 folding and defibrinogenating activities of acutobin recombinants from HEK293T 51:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 0 62 from HEK293T 122:128 arg1 recombinants 104:115 acutobin recombinants 95:115 acutobin recombinants from HEK293T 95:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 0 63 gly N-glycosylation 18:32 arg2 site 34:37 single N-glycosylation site knockout 11:46 single N-glycosylation site knockout 11:46 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 3 64 theme acutobin 438:445 arg1 mutants 415:421 single glycan-knockout mutants 392:421 single glycan-knockout mutants of recombinant acutobin 392:445 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 3 64 theme acutobin 438:445 arg1 ATB-wt 380:385 ATB-wt 380:385 ATB-wt 380:385 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 3 64 theme acutobin 438:445 arg1 wild-type 369:377 The wild-type 365:377 The wild-type (ATB-wt) 365:386 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 6 65 theme human 1203:1207 arg1 subunits 1220:1227 all human fibrinogen subunits 1199:1227 all human fibrinogen subunits faster but less specifically in vitro 1199:1265 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 5 66 dep conformation 1074:1085 arg1 the 1064:1066 the 1064:1066 the 1064:1066 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 7 67 theme defibrinogenating 1491:1507 arg1 stronger 1530:1537 stronger 1530:1537 stronger 1530:1537 In the mice model, the defibrinogenating effect of ATB-wt was stronger and lasting-longer than those of all the mutants. 25533529 7 67 theme defibrinogenating 1491:1507 arg1 effect 1509:1514 the defibrinogenating effect 1487:1514 the defibrinogenating effect of ATB-wt 1487:1524 In the mice model, the defibrinogenating effect of ATB-wt was stronger and lasting-longer than those of all the mutants. 25533529 3 68 theme various 584:590 arg1 mutants 616:622 various Asn(229)-deglycosylated mutants 584:622 various Asn(229)-deglycosylated mutants 584:622 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 8 69 theme fibrinogen-chain 1764:1779 arg1 specificity 1781:1791 the fibrinogen-chain specificity 1760:1791 the fibrinogen-chain specificity of acutobin 1760:1803 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 25533529 1 70 theme complex-typed 250:262 arg1 glycans 264:270 disialylated complex-typed glycans 237:270 disialylated complex-typed glycans 237:270 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 4 71 theme thrombin-like 737:749 arg1 enzymes 751:757 various venom thrombin-like enzymes 723:757 various venom thrombin-like enzymes 723:757 Based on homology modeling of acutobin and multiple sequence alignment with various venom thrombin-like enzymes, the importance of a hydrophilic environment at each glycosylation site to the enzyme folding could be rationalized. 25533529 3 72 theme Asn 592:594 arg1 mutants 616:622 various Asn(229)-deglycosylated mutants 584:622 various Asn(229)-deglycosylated mutants 584:622 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 2 73 theme site-directed 338:350 arg1 mutagenesis 352:362 site-directed mutagenesis 338:362 site-directed mutagenesis 338:362 Here, we explore the functional roles of each of the N-glycan by site-directed mutagenesis. 25533529 6 74 theme human 1420:1424 arg1 plasma 1426:1431 human plasma 1420:1431 human plasma slower than the other mutants did 1420:1465 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 0 75 theme defibrinogenating 63:79 arg1 activities 81:90 folding and defibrinogenating activities 51:90 folding and defibrinogenating activities of acutobin recombinants from HEK293T 51:128 Effects of single N-glycosylation site knockout on folding and defibrinogenating activities of acutobin recombinants from HEK293T. 25533529 5 76 theme sites 1115:1119 arg1 stability 1091:1099 stability 1091:1099 stability 1091:1099 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 5 76 theme sites 1115:1119 arg1 conformation 1074:1085 gross conformation 1068:1085 gross conformation 1068:1085 Remarkably, all the mutants showed similar catalytic activities for the chromogenic substrate and similar thermal stabilities as ATB-wt, suggesting that the glycan knockout did not affect the gross conformation and stability of the active sites. 25533529 6 77 theme SDS-PAGE 1131:1138 arg1 analyses 1140:1147 SDS-PAGE analyses 1131:1147 SDS-PAGE analyses 1131:1147 Although SDS-PAGE analyses revealed that ATB-wt and the D229-mutant degraded all human fibrinogen subunits faster but less specifically in vitro as compared with other mutants that cleaved only the α-subunit, ATB-wt and D229-mutant were not able to release fibrinogen-peptide A and thus coagulated human plasma slower than the other mutants did. 25533529 3 78 theme single 392:397 arg1 mutants 415:421 single glycan-knockout mutants 392:421 single glycan-knockout mutants of recombinant acutobin 392:445 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 1 79 from venom 189:193 arg1 Acutobin 131:138 Acutobin 131:138 Acutobin 131:138 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 1 79 from venom 189:193 arg1 α-fibrinogenase 145:159 the α-fibrinogenase 141:159 the α-fibrinogenase from Deinagkistrodon acutus venom 141:193 Acutobin, the α-fibrinogenase from Deinagkistrodon acutus venom, contains four N-glycosylation sites with disialylated complex-typed glycans. 25533529 3 80 theme glycan-knockout 399:413 arg1 mutants 415:421 single glycan-knockout mutants 392:421 single glycan-knockout mutants of recombinant acutobin 392:445 The wild-type (ATB-wt) and single glycan-knockout mutants of recombinant acutobin were prepared from HEK293T, demonstrating that mutations at Asn(77), Asn(81) and Asn(100) impaired the folding while the S79A mutant and various Asn(229)-deglycosylated mutants were correctly folded. 25533529 8 81 theme acutobin 1659:1666 arg1 pharmacokinetics 1639:1654 the pharmacokinetics 1635:1654 the pharmacokinetics of acutobin and ATB-wt in vivo 1635:1685 Taken together, all the glycans contribute to the pharmacokinetics of acutobin and ATB-wt in vivo, and the microenvironment around the Asn(229)-glycan appears to regulate the fibrinogen-chain specificity of acutobin while the N-glycans at positions 77, 81 and 100 are crucial for its folding. 26231935 3 0 dep AChE 671:674 arg1 i.e. 665:668 i.e. 665:668 i.e. 665:668 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 1 1 gly glycosylation 187:199 arg2 three 162:166 three 162:166 three 162:166 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 1 gly glycosylation 187:199 arg2 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 2 2 theme AChE 354:357 arg1 N-glycosylation 335:349 N-glycosylation 335:349 N-glycosylation of AChE 335:357 Several lines of evidence demonstrated that N-glycosylation of AChE affected the enzymatic activity, as well as its biosynthesis. 26231935 5 3 theme other 947:951 arg1 mutants 953:959 the other mutants 943:959 the other mutants 943:959 The mutants, AChE(T) (3N→3Q) and AChE(T) (N381Q), showed very minimal enzymatic activity, while the other mutants showed reduced activity. 26231935 4 4 theme protein 802:808 arg1 expression 810:819 AChE protein expression 797:819 AChE protein expression in the transfected cells 797:844 The mutation did not affect AChE protein expression in the transfected cells. 26231935 7 5 theme similar 1216:1222 arg1 composition 1231:1241 similar glycan composition 1216:1241 similar glycan composition 1216:1241 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 1 6 dep AChE 115:118 arg1 EC 121:122 EC 3.1.1.7 121:130 AChE; EC 3.1.1.7 115:130 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 7 7 theme glycan 1224:1229 arg1 composition 1231:1241 similar glycan composition 1216:1241 similar glycan composition 1216:1241 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 3 8 from activity 491:498 arg1 role 447:450 the role 443:450 the role of three N-glycosylation sites in AChE activity and glycan composition 443:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 7 9 theme N-glycosylation 1170:1184 arg1 mutants 1186:1192 various N-glycosylation mutants 1162:1192 various N-glycosylation mutants 1162:1192 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 7 10 gly N-glycosylation 1277:1291 arg2 sites 1293:1297 different N-glycosylation sites 1267:1297 different N-glycosylation sites 1267:1297 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 3 11 dep mutants 656:662 arg1 T 712:712 T 712:712 T 712:712 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 N381Q 695:699 N381Q 695:699 N381Q 695:699 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 N296Q 680:684 N296Q 680:684 N296Q 680:684 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 AChE 671:674 AChE 671:674 AChE(T) (N296Q) 671:685 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 N495Q 716:720 N495Q 716:720 N495Q 716:720 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 AChE 707:710 AChE 707:710 AChE 707:710 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 mutants 656:662 the single-site mutants 640:662 the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) 640:722 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 11 dep mutants 656:662 arg1 T 676:676 T 676:676 T 676:676 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 7 12 theme significant 1131:1141 arg1 difference 1143:1152 no significant difference 1128:1152 no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites 1128:1297 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 3 13 theme site-directed 528:540 arg1 mutagenesis 542:552 the site-directed mutagenesis 524:552 the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence 524:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 14 theme AChE 486:489 arg1 activity 491:498 AChE activity 486:498 AChE activity 486:498 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 15 theme N-glycosylation 461:475 arg1 sites 477:481 three N-glycosylation sites 455:481 three N-glycosylation sites in AChE activity and glycan composition 455:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 16 from mutagenesis 542:552 arg1 sequence 606:613 wild-type human AChE(T) sequence 582:613 wild-type human AChE(T) sequence 582:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 6 17 theme glycosylation 1029:1041 arg1 profile 1043:1049 the glycosylation profile 1025:1049 the glycosylation profile 1025:1049 By binding to lectins, Con A, and SNA, the glycosylation profile was revealed in those mutated AChE. 26231935 3 18 dep mutant 737:742 arg1 T 756:756 T 756:756 T 756:756 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 18 dep mutant 737:742 arg1 3N→3Q 760:764 3N→3Q 760:764 3N→3Q 760:764 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 18 dep mutant 737:742 arg1 AChE 751:754 AChE 751:754 AChE(T) (3N→3Q) 751:765 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 8 19 gly glycosylation 1319:1331 arg2 sites 1333:1337 the three glycosylation sites 1309:1337 the three glycosylation sites within AChE sequence 1309:1358 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 8 19 gly glycosylation 1319:1331 arg2 three 1313:1317 three 1313:1317 three 1313:1317 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 1 20 theme N-linked 178:185 arg1 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 5 21 theme reduced 968:974 arg1 activity 976:983 reduced activity 968:983 reduced activity 968:983 The mutants, AChE(T) (3N→3Q) and AChE(T) (N381Q), showed very minimal enzymatic activity, while the other mutants showed reduced activity. 26231935 1 22 contain possessing 151:160 arg1 Acetylcholinesterase 93:112 Acetylcholinesterase 93:112 Acetylcholinesterase (AChE; EC 3.1.1.7) 93:131 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 22 contain possessing 151:160 arg1 glycoprotein 138:149 a glycoprotein 136:149 a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species 136:226 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 22 contain possessing 151:160 arg2 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 8 23 theme enzymatic 1399:1407 arg1 activity 1409:1416 the enzymatic activity 1395:1416 the enzymatic activity 1395:1416 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 7 24 theme various 1162:1168 arg1 mutants 1186:1192 various N-glycosylation mutants 1162:1192 various N-glycosylation mutants 1162:1192 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 1 25 theme glycosylation 187:199 arg1 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 0 26 theme N-Glycosylation 6:20 arg1 Sites 22:26 Three N-Glycosylation Sites 0:26 Three N-Glycosylation Sites of Human Acetylcholinesterase 0:56 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 0 26 theme N-Glycosylation 6:20 arg1 Acetylcholinesterase 37:56 Human Acetylcholinesterase 31:56 Human Acetylcholinesterase 31:56 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 8 27 theme glycosylation 1319:1331 arg1 sites 1333:1337 the three glycosylation sites 1309:1337 the three glycosylation sites within AChE sequence 1309:1358 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 1 28 link N-linked 178:185 arg1 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 3 29 theme N-glycosylation 557:571 arg1 sites 573:577 N-glycosylation sites 557:577 N-glycosylation sites 557:577 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 1 30 gly glycoprotein 138:149 arg1 Acetylcholinesterase 93:112 Acetylcholinesterase 93:112 Acetylcholinesterase (AChE; EC 3.1.1.7) 93:131 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 30 gly glycoprotein 138:149 arg1 glycoprotein 138:149 a glycoprotein 136:149 a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species 136:226 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 6 31 theme Con 1009:1011 arg1 A 1013:1013 Con A 1009:1013 Con A 1009:1013 By binding to lectins, Con A, and SNA, the glycosylation profile was revealed in those mutated AChE. 26231935 3 32 from sites 477:481 arg1 activity 491:498 AChE activity 486:498 AChE activity 486:498 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 32 from sites 477:481 arg1 composition 511:521 glycan composition 504:521 glycan composition 504:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 33 theme sites 573:577 arg1 mutagenesis 542:552 the site-directed mutagenesis 524:552 the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence 524:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 2 34 theme enzymatic 372:380 arg1 activity 382:389 the enzymatic activity 368:389 the enzymatic activity 368:389 Several lines of evidence demonstrated that N-glycosylation of AChE affected the enzymatic activity, as well as its biosynthesis. 26231935 3 35 dep AChE 751:754 arg1 i.e. 745:748 i.e. 745:748 i.e. 745:748 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 2 36 theme Several 291:297 arg1 lines 299:303 Several lines 291:303 Several lines of evidence 291:315 Several lines of evidence demonstrated that N-glycosylation of AChE affected the enzymatic activity, as well as its biosynthesis. 26231935 1 37 theme mammalian 210:218 arg1 species 220:226 mammalian species 210:226 mammalian species 210:226 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 38 theme human 275:279 arg1 sequence 281:288 the human sequence 271:288 the human sequence 271:288 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 8 39 theme glycan 1425:1430 arg1 compositions 1432:1443 their glycan compositions 1419:1443 their glycan compositions 1419:1443 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 1 40 theme conserved 168:176 arg1 sites 201:205 three conserved N-linked glycosylation sites 162:205 three conserved N-linked glycosylation sites 162:205 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 0 41 theme Acetylcholinesterase 37:56 arg1 Sites 22:26 Three N-Glycosylation Sites 0:26 Three N-Glycosylation Sites of Human Acetylcholinesterase 0:56 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 0 41 theme Acetylcholinesterase 37:56 arg1 Acetylcholinesterase 37:56 Human Acetylcholinesterase 31:56 Human Acetylcholinesterase 31:56 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 1 42 theme sequence 281:288 arg1 sequence 281:288 the human sequence 271:288 the human sequence 271:288 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 1 42 theme sequence 281:288 arg1 residues 259:266 296, 381, and 495 residues 241:266 296, 381, and 495 residues of the human sequence 241:288 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 3 43 theme human 592:596 arg1 sequence 606:613 wild-type human AChE(T) sequence 582:613 wild-type human AChE(T) sequence 582:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 0 44 theme Human 31:35 arg1 Acetylcholinesterase 37:56 Human Acetylcholinesterase 31:56 Human Acetylcholinesterase 31:56 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 8 45 theme AChE 1346:1349 arg1 sequence 1351:1358 AChE sequence 1346:1358 AChE sequence 1346:1358 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 3 46 gly N-glycosylation 557:571 arg2 sites 573:577 N-glycosylation sites 557:577 N-glycosylation sites 557:577 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 7 47 theme N-glycosylation 1277:1291 arg1 sites 1293:1297 different N-glycosylation sites 1267:1297 different N-glycosylation sites 1267:1297 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 3 48 theme AChE 598:601 arg1 sequence 606:613 wild-type human AChE(T) sequence 582:613 wild-type human AChE(T) sequence 582:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 6 49 theme mutated 1073:1079 arg1 AChE 1081:1084 those mutated AChE 1067:1084 those mutated AChE 1067:1084 By binding to lectins, Con A, and SNA, the glycosylation profile was revealed in those mutated AChE. 26231935 0 50 theme Similar 65:71 arg1 Composition 80:90 Similar Glycan Composition 65:90 Similar Glycan Composition 65:90 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 7 51 theme binding 1091:1097 arg1 affinity 1099:1106 The binding affinity 1087:1106 The binding affinity with lectins 1087:1119 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 8 52 contain have 1360:1363 arg1 sites 1333:1337 the three glycosylation sites 1309:1337 the three glycosylation sites within AChE sequence 1309:1358 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 8 52 contain have 1360:1363 arg2 extent 1375:1380 different extent 1365:1380 different extent 1365:1380 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 3 53 theme sites 477:481 arg1 role 447:450 the role 443:450 the role of three N-glycosylation sites in AChE activity and glycan composition 443:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 4 54 theme transfected 828:838 arg1 cells 840:844 the transfected cells 824:844 the transfected cells 824:844 The mutation did not affect AChE protein expression in the transfected cells. 26231935 5 55 theme minimal 909:915 arg1 activity 927:934 very minimal enzymatic activity 904:934 very minimal enzymatic activity 904:934 The mutants, AChE(T) (3N→3Q) and AChE(T) (N381Q), showed very minimal enzymatic activity, while the other mutants showed reduced activity. 26231935 2 56 gly N-glycosylation 335:349 arg1 AChE 354:357 AChE 354:357 AChE 354:357 Several lines of evidence demonstrated that N-glycosylation of AChE affected the enzymatic activity, as well as its biosynthesis. 26231935 3 57 theme wild-type 582:590 arg1 sequence 606:613 wild-type human AChE(T) sequence 582:613 wild-type human AChE(T) sequence 582:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 58 from role 447:450 arg1 activity 491:498 AChE activity 486:498 AChE activity 486:498 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 58 from role 447:450 arg1 composition 511:521 glycan composition 504:521 glycan composition 504:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 1 59 dep Acetylcholinesterase 93:112 arg1 AChE 115:118 AChE 115:118 AChE; EC 3.1.1.7 115:130 Acetylcholinesterase (AChE; EC 3.1.1.7) is a glycoprotein possessing three conserved N-linked glycosylation sites in mammalian species, locating at 296, 381, and 495 residues of the human sequence. 26231935 3 60 theme site 732:735 arg1 mutant 737:742 all site mutant 728:742 all site mutant (i.e., AChE(T) (3N→3Q)) 728:766 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 5 61 theme enzymatic 917:925 arg1 activity 927:934 very minimal enzymatic activity 904:934 very minimal enzymatic activity 904:934 The mutants, AChE(T) (3N→3Q) and AChE(T) (N381Q), showed very minimal enzymatic activity, while the other mutants showed reduced activity. 26231935 7 62 theme different 1267:1275 arg1 sites 1293:1297 different N-glycosylation sites 1267:1297 different N-glycosylation sites 1267:1297 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 26231935 3 63 gly N-glycosylation 461:475 arg2 three 455:459 three 455:459 three 455:459 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 63 gly N-glycosylation 461:475 arg2 sites 477:481 three N-glycosylation sites 455:481 three N-glycosylation sites in AChE activity and glycan composition 455:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 4 64 from expression 810:819 arg1 cells 840:844 the transfected cells 824:844 the transfected cells 824:844 The mutation did not affect AChE protein expression in the transfected cells. 26231935 2 65 theme evidence 308:315 arg1 lines 299:303 Several lines 291:303 Several lines of evidence 291:315 Several lines of evidence demonstrated that N-glycosylation of AChE affected the enzymatic activity, as well as its biosynthesis. 26231935 8 66 theme different 1365:1373 arg1 extent 1375:1380 different extent 1365:1380 different extent 1365:1380 Although the three glycosylation sites within AChE sequence have different extent in affecting the enzymatic activity, their glycan compositions are very similar. 26231935 4 67 theme AChE 797:800 arg1 expression 810:819 AChE protein expression 797:819 AChE protein expression in the transfected cells 797:844 The mutation did not affect AChE protein expression in the transfected cells. 26231935 3 68 theme single-site 644:654 arg1 AChE 671:674 AChE 671:674 AChE(T) (N296Q) 671:685 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 68 theme single-site 644:654 arg1 AChE 707:710 AChE 707:710 AChE 707:710 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 68 theme single-site 644:654 arg1 mutants 656:662 the single-site mutants 640:662 the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) 640:722 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 68 theme single-site 644:654 arg1 N381Q 695:699 N381Q 695:699 N381Q 695:699 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 0 69 theme Glycan 73:78 arg1 Composition 80:90 Similar Glycan Composition 65:90 Similar Glycan Composition 65:90 Three N-Glycosylation Sites of Human Acetylcholinesterase Shares Similar Glycan Composition. 26231935 3 70 theme T 603:603 arg1 sequence 606:613 wild-type human AChE(T) sequence 582:613 wild-type human AChE(T) sequence 582:613 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 71 theme glycan 504:509 arg1 composition 511:521 glycan composition 504:521 glycan composition 504:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 3 72 from composition 511:521 arg1 role 447:450 the role 443:450 the role of three N-glycosylation sites in AChE activity and glycan composition 443:521 In order to determine the role of three N-glycosylation sites in AChE activity and glycan composition, the site-directed mutagenesis of N-glycosylation sites in wild-type human AChE(T) sequence was employed to generate the single-site mutants (i.e., AChE(T) (N296Q), AChET (N381Q), and AChE(T) (N495Q)) and all site mutant (i.e., AChE(T) (3N→3Q)). 26231935 7 73 with affinity 1099:1106 arg1 lectins 1113:1119 lectins 1113:1119 lectins 1113:1119 The binding affinity with lectins showed no significant difference between various N-glycosylation mutants, which suggested that similar glycan composition should be resulted from different N-glycosylation sites. 28516782 1 0 theme proteins 277:284 arg1 relationships 229:241 structure-function relationships 210:241 structure-function relationships of these extensively glycosylated proteins 210:284 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 8 1 theme hinge 1248:1252 arg1 region 1254:1259 the proline-rich hinge region 1231:1259 the proline-rich hinge region from HEK293-6E cell-derived IgA1 1231:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 6 2 from IgAs 955:958 arg1 contrary 968:975 the contrary 964:975 the contrary 964:975 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 7 3 theme individual 1167:1176 arg1 sites 1194:1198 the individual N-glycosylation sites 1163:1198 the individual N-glycosylation sites of each IgA subtype 1163:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 7 3 theme individual 1167:1176 arg1 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 3 4 theme IgA 513:515 arg1 isotypes 517:524 All three IgA isotypes 503:524 All three IgA isotypes 503:524 All three IgA isotypes were purified and subjected to biophysical and biochemical characterization. 28516782 5 5 theme complex-type 900:911 arg1 N-glycans 925:933 mainly complex-type biantennary N-glycans 893:933 mainly complex-type biantennary N-glycans 893:933 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 8 6 from IgA1 1289:1292 arg1 region 1254:1259 the proline-rich hinge region 1231:1259 the proline-rich hinge region from HEK293-6E cell-derived IgA1 1231:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 4 7 theme glycosylation 658:670 arg1 occupancy 672:680 glycosylation occupancy 658:680 glycosylation occupancy 658:680 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 8 8 theme cell-derived 1276:1287 arg1 IgA1 1289:1292 HEK293-6E cell-derived IgA1 1266:1292 HEK293-6E cell-derived IgA1 1266:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 6 9 with N-glycans 1004:1012 arg1 levels 1024:1029 high levels 1019:1029 high levels of sialylation, core-fucose, and the presence of branched structures 1019:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 7 10 theme major 1137:1141 arg1 differences 1143:1153 major differences 1137:1153 major differences between the individual N-glycosylation sites of each IgA subtype 1137:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 2 11 theme monomeric 316:324 arg1 IgA1 326:329 Here monomeric IgA1 311:329 Here monomeric IgA1 311:329 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 5 12 theme plant-produced 867:880 arg1 IgAs 882:885 plant-produced IgAs 867:885 plant-produced IgAs 867:885 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 4 13 theme antigen 637:643 arg1 binding 645:651 antigen binding 637:651 antigen binding 637:651 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 9 14 theme distinct 1590:1597 arg1 glycoforms 1599:1608 distinct glycoforms 1590:1608 distinct glycoforms 1590:1608 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 6 15 theme heterogeneous 990:1002 arg1 N-glycans 1004:1012 very heterogeneous N-glycans 985:1012 very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures 985:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 8 16 used occupied 1298:1305 arg2 region 1254:1259 the proline-rich hinge region 1231:1259 the proline-rich hinge region from HEK293-6E cell-derived IgA1 1231:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 8 17 theme numerous 1377:1384 arg1 modifications 1401:1413 numerous plant-specific modifications 1377:1413 numerous plant-specific modifications 1377:1413 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 1 18 theme A 138:138 arg1 potential 98:106 The full potential 89:106 The full potential of recombinant Immunoglobulin A as therapeutic antibody 89:162 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 7 19 theme subtype 1212:1218 arg1 sites 1194:1198 the individual N-glycosylation sites 1163:1198 the individual N-glycosylation sites of each IgA subtype 1163:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 7 19 theme subtype 1212:1218 arg1 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 8 20 from benthamiana 1355:1365 arg1 IgA1 1342:1345 IgA1 1342:1345 IgA1 from N. benthamiana 1342:1365 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 9 21 theme thermal 1621:1627 arg1 stability 1629:1637 the thermal stability 1617:1637 the thermal stability of IgAs 1617:1645 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 6 22 theme presence 1068:1075 arg1 levels 1024:1029 high levels 1019:1029 high levels of sialylation, core-fucose, and the presence of branched structures 1019:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 2 23 theme human 480:484 arg1 cells 496:500 human HEK293-6E cells 480:500 human HEK293-6E cells 480:500 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 0 24 theme IgA 75:77 arg1 Isotypes 79:86 Plant-Produced Human IgA Isotypes 54:86 Plant-Produced Human IgA Isotypes 54:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 8 25 link cell-derived 1276:1287 arg1 IgA1 1289:1292 HEK293-6E cell-derived IgA1 1266:1292 HEK293-6E cell-derived IgA1 1266:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 6 26 theme core-fucose 1047:1057 arg1 levels 1024:1029 high levels 1019:1029 high levels of sialylation, core-fucose, and the presence of branched structures 1019:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 6 27 theme sialylation 1034:1044 arg1 levels 1024:1029 high levels 1019:1029 high levels of sialylation, core-fucose, and the presence of branched structures 1019:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 1 28 theme therapeutic 143:153 arg1 antibody 155:162 therapeutic antibody 143:162 therapeutic antibody 143:162 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 0 29 theme Isotypes 79:86 arg1 N-Glycosylation 24:38 Site-Specific N-Glycosylation 10:38 Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes 10:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 5 30 theme site-specific 825:837 arg1 glycosylation 839:851 site-specific glycosylation 825:851 site-specific glycosylation 825:851 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 2 31 theme trastuzumab 397:407 arg1 variants 355:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants 311:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab 311:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 4 32 theme glycan 740:745 arg1 structures 747:756 glycan structures 740:756 glycan structures 740:756 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 2 33 theme IgG1 391:394 arg1 trastuzumab 397:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 1 34 theme recombinant 111:121 arg1 A 138:138 recombinant Immunoglobulin A 111:138 recombinant Immunoglobulin A 111:138 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 2 35 theme benthamiana 454:464 arg1 plants 466:471 glyco-engineered Nicotiana benthamiana plants 427:471 glyco-engineered Nicotiana benthamiana plants 427:471 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 6 36 theme structures 1089:1098 arg1 core-fucose 1047:1057 core-fucose 1047:1057 core-fucose 1047:1057 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 6 36 theme structures 1089:1098 arg1 sialylation 1034:1044 sialylation 1034:1044 sialylation 1034:1044 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 6 36 theme structures 1089:1098 arg1 presence 1068:1075 the presence 1064:1075 the presence of branched structures 1064:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 2 37 theme antibody 381:388 arg1 trastuzumab 397:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 0 38 theme HEK293 43:48 arg1 N-Glycosylation 24:38 Site-Specific N-Glycosylation 10:38 Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes 10:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 6 39 theme branched 1080:1087 arg1 structures 1089:1098 branched structures 1080:1098 branched structures 1080:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 2 40 theme glyco-engineered 427:442 arg1 plants 466:471 glyco-engineered Nicotiana benthamiana plants 427:471 glyco-engineered Nicotiana benthamiana plants 427:471 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 0 41 theme Plant-Produced 54:67 arg1 Isotypes 79:86 Plant-Produced Human IgA Isotypes 54:86 Plant-Produced Human IgA Isotypes 54:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 9 42 theme domain 1463:1468 arg1 unfolding 1442:1450 unfolding 1442:1450 unfolding of the CH2 domain of plant-produced IgA toward lower temperatures 1442:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 9 43 theme plant-produced 1473:1486 arg1 IgA 1488:1490 plant-produced IgA 1473:1490 plant-produced IgA toward lower temperatures 1473:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 9 44 theme differential 1539:1550 arg1 calorimetry 1561:1571 differential scanning calorimetry 1539:1571 differential scanning calorimetry 1539:1571 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 0 45 theme Human 69:73 arg1 Isotypes 79:86 Plant-Produced Human IgA Isotypes 54:86 Plant-Produced Human IgA Isotypes 54:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 2 46 theme IgA2m 346:350 arg1 variants 355:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants 311:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab 311:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 1 47 theme glycosylated 264:275 arg1 proteins 277:284 these extensively glycosylated proteins 246:284 these extensively glycosylated proteins 246:284 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 7 48 theme N-glycosylation 1178:1192 arg1 sites 1194:1198 the individual N-glycosylation sites 1163:1198 the individual N-glycosylation sites of each IgA subtype 1163:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 7 48 theme N-glycosylation 1178:1192 arg1 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 9 49 theme scanning 1552:1559 arg1 calorimetry 1561:1571 differential scanning calorimetry 1539:1571 differential scanning calorimetry 1539:1571 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 8 50 theme proline-rich 1235:1246 arg1 region 1254:1259 the proline-rich hinge region 1231:1259 the proline-rich hinge region from HEK293-6E cell-derived IgA1 1231:1292 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 6 51 theme HEK293-6E-produced 936:953 arg1 IgAs 955:958 HEK293-6E-produced IgAs 936:958 HEK293-6E-produced IgAs 936:958 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 8 52 theme mucin-type 1312:1321 arg1 O-glycans 1323:1331 mucin-type O-glycans 1312:1331 mucin-type O-glycans 1312:1331 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 2 53 theme IgA2m 332:336 arg1 variants 355:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants 311:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab 311:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 5 54 theme biantennary 913:923 arg1 N-glycans 925:933 mainly complex-type biantennary N-glycans 893:933 mainly complex-type biantennary N-glycans 893:933 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 2 55 theme IgA1 326:329 arg1 variants 355:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants 311:362 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab 311:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 6 56 theme high 1019:1022 arg1 levels 1024:1029 high levels 1019:1029 high levels of sialylation, core-fucose, and the presence of branched structures 1019:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 7 57 theme site-specific 1105:1117 arg1 analysis 1119:1126 The site-specific analysis 1101:1126 The site-specific analysis 1101:1126 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 3 58 theme biochemical 573:583 arg1 characterization 585:600 biophysical and biochemical characterization 557:600 biophysical and biochemical characterization 557:600 All three IgA isotypes were purified and subjected to biophysical and biochemical characterization. 28516782 7 59 gly N-glycosylation 1178:1192 arg2 sites 1194:1198 the individual N-glycosylation sites 1163:1198 the individual N-glycosylation sites of each IgA subtype 1163:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 7 59 gly N-glycosylation 1178:1192 arg1 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 7 59 gly N-glycosylation 1178:1192 arg2 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 3 60 theme biophysical 557:567 arg1 characterization 585:600 biophysical and biochemical characterization 557:600 biophysical and biochemical characterization 557:600 All three IgA isotypes were purified and subjected to biophysical and biochemical characterization. 28516782 1 61 theme Immunoglobulin 123:136 arg1 A 138:138 recombinant Immunoglobulin A 111:138 recombinant Immunoglobulin A 111:138 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 8 62 theme plant-specific 1386:1399 arg1 modifications 1401:1413 numerous plant-specific modifications 1377:1413 numerous plant-specific modifications 1377:1413 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 4 63 from differences 612:622 arg1 occupancy 672:680 glycosylation occupancy 658:680 glycosylation occupancy 658:680 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 4 63 from differences 612:622 arg1 assembly 627:634 assembly 627:634 assembly 627:634 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 4 63 from differences 612:622 arg1 binding 645:651 antigen binding 637:651 antigen binding 637:651 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 4 64 gly heterogeneity 762:774 arg1 glycosylation 779:791 glycosylation 779:791 glycosylation 779:791 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 9 65 theme IgAs 1642:1645 arg1 stability 1629:1637 the thermal stability 1617:1637 the thermal stability of IgAs 1617:1645 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 4 66 theme structures 747:756 arg1 terms 731:735 terms 731:735 terms of glycan structures and heterogeneity of glycosylation 731:791 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 2 67 theme HEK293-6E 486:494 arg1 cells 496:500 human HEK293-6E cells 480:500 human HEK293-6E cells 480:500 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 5 68 contain carry 887:891 arg1 IgAs 882:885 plant-produced IgAs 867:885 plant-produced IgAs 867:885 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 5 68 contain carry 887:891 arg2 N-glycans 925:933 mainly complex-type biantennary N-glycans 893:933 mainly complex-type biantennary N-glycans 893:933 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 4 69 theme heterogeneity 762:774 arg1 terms 731:735 terms 731:735 terms of glycan structures and heterogeneity of glycosylation 731:791 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 0 70 theme Site-Specific 10:22 arg1 N-Glycosylation 24:38 Site-Specific N-Glycosylation 10:38 Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes 10:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 9 71 from shift 1433:1437 arg1 unfolding 1442:1450 unfolding 1442:1450 unfolding of the CH2 domain of plant-produced IgA toward lower temperatures 1442:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 1 72 gly glycosylated 264:275 arg1 proteins 277:284 these extensively glycosylated proteins 246:284 these extensively glycosylated proteins 246:284 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 1 73 theme full 93:96 arg1 potential 98:106 The full potential 89:106 The full potential of recombinant Immunoglobulin A as therapeutic antibody 89:162 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 5 74 theme glycosylation 839:851 arg1 analysis 813:820 Mass-spectrometric analysis 794:820 Mass-spectrometric analysis of site-specific glycosylation 794:851 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 7 75 theme IgA 1208:1210 arg1 subtype 1212:1218 each IgA subtype 1203:1218 each IgA subtype 1203:1218 The site-specific analysis revealed major differences between the individual N-glycosylation sites of each IgA subtype. 28516782 2 76 theme Nicotiana 444:452 arg1 plants 466:471 glyco-engineered Nicotiana benthamiana plants 427:471 glyco-engineered Nicotiana benthamiana plants 427:471 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 2 77 theme anti-HER2 371:379 arg1 trastuzumab 397:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 the anti-HER2 antibody (IgG1) trastuzumab 367:407 Here monomeric IgA1, IgA2m(1), and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. 28516782 5 78 theme Mass-spectrometric 794:811 arg1 analysis 813:820 Mass-spectrometric analysis 794:820 Mass-spectrometric analysis of site-specific glycosylation 794:851 Mass-spectrometric analysis of site-specific glycosylation revealed that plant-produced IgAs carry mainly complex-type biantennary N-glycans. 28516782 9 79 theme CH2 1459:1461 arg1 domain 1463:1468 the CH2 domain 1455:1468 the CH2 domain of plant-produced IgA toward lower temperatures 1455:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 9 79 theme CH2 1459:1461 arg1 IgA 1488:1490 plant-produced IgA 1473:1490 plant-produced IgA toward lower temperatures 1473:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 0 80 gly N-Glycosylation 24:38 arg1 Isotypes 79:86 Plant-Produced Human IgA Isotypes 54:86 Plant-Produced Human IgA Isotypes 54:86 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 0 80 gly N-Glycosylation 24:38 arg1 HEK293 43:48 HEK293 43:48 HEK293 43:48 Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes. 28516782 6 81 gly sialylation 1034:1044 arg1 structures 1089:1098 branched structures 1080:1098 branched structures 1080:1098 HEK293-6E-produced IgAs, on the contrary, showed very heterogeneous N-glycans with high levels of sialylation, core-fucose, and the presence of branched structures. 28516782 8 82 theme N. 1352:1353 arg1 benthamiana 1355:1365 N. benthamiana 1352:1365 N. benthamiana 1352:1365 Moreover, the proline-rich hinge region from HEK293-6E cell-derived IgA1 was occupied with mucin-type O-glycans, whereas IgA1 from N. benthamiana displayed numerous plant-specific modifications. 28516782 4 83 theme glycosylation 779:791 arg1 heterogeneity 762:774 heterogeneity 762:774 heterogeneity of glycosylation 762:791 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 4 83 theme glycosylation 779:791 arg1 structures 747:756 glycan structures 740:756 glycan structures 740:756 While no differences in assembly, antigen binding, and glycosylation occupancy were observed, both systems vary tremendously in terms of glycan structures and heterogeneity of glycosylation. 28516782 9 84 theme IgA 1488:1490 arg1 domain 1463:1468 the CH2 domain 1455:1468 the CH2 domain of plant-produced IgA toward lower temperatures 1455:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 9 84 theme IgA 1488:1490 arg1 IgA 1488:1490 plant-produced IgA 1473:1490 plant-produced IgA toward lower temperatures 1473:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 28516782 1 85 theme structure-function 210:227 arg1 relationships 229:241 structure-function relationships 210:241 structure-function relationships of these extensively glycosylated proteins 210:284 The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. 28516782 9 86 theme lower 1499:1503 arg1 temperatures 1505:1516 lower temperatures 1499:1516 lower temperatures 1499:1516 Interestingly, a shift in unfolding of the CH2 domain of plant-produced IgA toward lower temperatures can be observed with differential scanning calorimetry, suggesting that distinct glycoforms affect the thermal stability of IgAs. 25628020 2 0 gly N-glycosylation 296:310 arg2 site 312:315 a novel N-glycosylation site 288:315 a novel N-glycosylation site 288:315 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 6 1 theme T4 1228:1229 arg1 production 1231:1240 T4 production 1228:1240 T4 production 1228:1240 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 6 2 theme thyroid 1065:1071 arg1 hormones 1073:1080 thyroid hormones 1065:1080 thyroid hormones 1065:1080 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 7 3 theme N-glycosylation 1282:1296 arg1 sites 1298:1302 the novel N-glycosylation sites 1272:1302 the novel N-glycosylation sites described in this study 1272:1326 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 7 3 theme N-glycosylation 1282:1296 arg1 candidates 1342:1351 useful candidates 1335:1351 useful candidates for functional analyses 1335:1375 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 0 4 theme human 76:80 arg1 evolution 82:90 human evolution 76:90 human evolution 76:90 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 0 5 from acquisition 4:14 arg1 proteins 60:67 conserved proteins 50:67 conserved proteins 50:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 7 6 theme human 1464:1468 arg1 lineage 1470:1476 the human lineage 1460:1476 the human lineage 1460:1476 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 1 7 theme various 163:169 arg1 signaling 258:266 signaling 258:266 signaling 258:266 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 7 theme various 163:169 arg1 trafficking 223:233 trafficking 223:233 trafficking 223:233 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 7 theme various 163:169 arg1 adhesion 245:252 cell adhesion 240:252 cell adhesion 240:252 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 7 theme various 163:169 arg1 processes 182:190 various biological processes 163:190 various biological processes 163:190 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 7 theme various 163:169 arg1 folding 211:217 protein folding 203:217 protein folding 203:217 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 3 8 dep Euarchonta 643:652 arg1 treeshrews 668:677 treeshrews 668:677 treeshrews 668:677 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 3 8 dep Euarchonta 643:652 arg1 primates 655:662 primates 655:662 primates 655:662 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 6 9 from production 1051:1060 arg1 humans 1085:1090 humans 1085:1090 humans 1085:1090 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 6 9 from production 1051:1060 arg1 thyroxine 1104:1112 thyroxine 1104:1112 especially thyroxine (T4) 1093:1117 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 2 10 theme novel 290:294 arg1 site 312:315 a novel N-glycosylation site 288:315 a novel N-glycosylation site 288:315 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 2 11 theme N-glycosylation 296:310 arg1 site 312:315 a novel N-glycosylation site 288:315 a novel N-glycosylation site 288:315 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 4 12 theme membrane-associated 724:742 arg1 protein 744:750 adipocyte plasma membrane-associated protein 707:750 adipocyte plasma membrane-associated protein (APMAP) 707:758 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 12 theme membrane-associated 724:742 arg1 APMAP 753:757 APMAP 753:757 APMAP 753:757 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 13 from protein 744:750 arg1 cluster 771:777 cluster 771:777 cluster of differentiation 166 (CD166/ALCAM) 771:814 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 13 from protein 744:750 arg1 thyroglobulin 831:843 thyroglobulin 831:843 thyroglobulin 831:843 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 3 14 theme novel 534:538 arg1 sites 556:560 112 novel N-glycosylation sites 530:560 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews) 530:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 3 15 gly N-glycosylation 540:554 arg2 sites 556:560 112 novel N-glycosylation sites 530:560 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews) 530:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 3 15 gly N-glycosylation 540:554 arg2 112 530:532 112 530:532 112 530:532 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 2 16 theme protein 348:354 arg1 structure 356:364 protein structure 348:364 protein structure 348:364 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 7 17 theme functional 1357:1366 arg1 analyses 1368:1375 functional analyses 1357:1375 functional analyses 1357:1375 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 1 18 theme biological 171:180 arg1 signaling 258:266 signaling 258:266 signaling 258:266 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 18 theme biological 171:180 arg1 trafficking 223:233 trafficking 223:233 trafficking 223:233 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 18 theme biological 171:180 arg1 adhesion 245:252 cell adhesion 240:252 cell adhesion 240:252 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 18 theme biological 171:180 arg1 processes 182:190 various biological processes 163:190 various biological processes 163:190 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 18 theme biological 171:180 arg1 folding 211:217 protein folding 203:217 protein folding 203:217 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 0 19 from sites 41:45 arg1 proteins 60:67 conserved proteins 50:67 conserved proteins 50:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 3 20 theme N-glycosylation 474:488 arg1 sites 490:494 2,534 N-glycosylation sites 468:494 2,534 N-glycosylation sites in 1,027 proteins 468:512 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 6 21 theme increased 1041:1049 arg1 production 1051:1060 the increased production 1037:1060 the increased production of thyroid hormones in humans, especially thyroxine (T4) 1037:1117 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 7 22 theme genetic 1400:1406 arg1 modifications 1408:1420 innovative genetic modifications 1389:1420 innovative genetic modifications for beneficial phenotypes acquired in the human lineage 1389:1476 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 4 23 theme plasma 717:722 arg1 protein 744:750 adipocyte plasma membrane-associated protein 707:750 adipocyte plasma membrane-associated protein (APMAP) 707:758 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 23 theme plasma 717:722 arg1 APMAP 753:757 APMAP 753:757 APMAP 753:757 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 24 from Asn-76 821:826 arg1 cluster 771:777 cluster 771:777 cluster of differentiation 166 (CD166/ALCAM) 771:814 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 24 from Asn-76 821:826 arg1 thyroglobulin 831:843 thyroglobulin 831:843 thyroglobulin 831:843 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 25 theme them 690:693 arg1 Asn-196 696:702 Asn-196 696:702 Asn-196 696:702 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 25 theme them 690:693 arg1 them 690:693 them 690:693 them 690:693 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 25 theme them 690:693 arg1 Three 681:685 Three 681:685 Three 681:685 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 3 26 gly N-glycosylation 474:488 arg2 sites 490:494 2,534 N-glycosylation sites 468:494 2,534 N-glycosylation sites in 1,027 proteins 468:512 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 3 27 from sites 556:560 arg1 proteins 568:575 91 proteins 565:575 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews) 565:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 0 28 theme novel 19:23 arg1 sites 41:45 novel N-glycosylation sites 19:45 novel N-glycosylation sites in conserved proteins 19:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 7 29 theme novel 1276:1280 arg1 sites 1298:1302 the novel N-glycosylation sites 1272:1302 the novel N-glycosylation sites described in this study 1272:1326 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 7 29 theme novel 1276:1280 arg1 candidates 1342:1351 useful candidates 1335:1351 useful candidates for functional analyses 1335:1375 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 3 30 from sites 490:494 arg1 proteins 505:512 1,027 proteins 499:512 1,027 proteins 499:512 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 2 31 theme site 312:315 arg1 acquisition 273:283 The acquisition 269:283 The acquisition of a novel N-glycosylation site 269:315 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 1 32 theme protein 203:209 arg1 folding 211:217 protein folding 203:217 protein folding 203:217 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 4 33 from Asn-91 761:766 arg1 cluster 771:777 cluster 771:777 cluster of differentiation 166 (CD166/ALCAM) 771:814 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 33 from Asn-91 761:766 arg1 thyroglobulin 831:843 thyroglobulin 831:843 thyroglobulin 831:843 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 3 34 theme common 624:629 arg1 ancestor 631:638 the last common ancestor 615:638 the last common ancestor of Euarchonta (primates and treeshrews) 615:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 1 35 theme N-linked 105:112 arg1 glycosylation 122:134 N-linked protein glycosylation 105:134 N-linked protein glycosylation 105:134 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 0 36 theme sites 41:45 arg1 acquisition 4:14 The acquisition 0:14 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution 0:90 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 3 37 theme human 438:442 arg1 set 463:465 the human glycoproteome data set 434:465 the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) 434:513 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 0 38 theme N-glycosylation 25:39 arg1 sites 41:45 novel N-glycosylation sites 19:45 novel N-glycosylation sites in conserved proteins 19:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 7 39 theme beneficial 1426:1435 arg1 phenotypes 1437:1446 beneficial phenotypes 1426:1446 beneficial phenotypes acquired in the human lineage 1426:1476 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 7 40 gly N-glycosylation 1282:1296 arg2 candidates 1342:1351 useful candidates 1335:1351 useful candidates for functional analyses 1335:1375 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 7 40 gly N-glycosylation 1282:1296 arg2 sites 1298:1302 the novel N-glycosylation sites 1272:1302 the novel N-glycosylation sites described in this study 1272:1326 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 3 41 theme glycoproteome 444:456 arg1 set 463:465 the human glycoproteome data set 434:465 the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) 434:513 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 0 42 theme conserved 50:58 arg1 proteins 60:67 conserved proteins 50:67 conserved proteins 50:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 6 43 theme thyroglobulin 1002:1014 arg1 Asn-76 992:997 the Asn-76 988:997 the Asn-76 of thyroglobulin 988:1014 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 0 44 from proteins 60:67 arg1 acquisition 4:14 The acquisition 0:14 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution 0:90 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 5 45 theme positive 936:943 arg1 selection 945:953 positive selection 936:953 positive selection 936:953 Molecular evolutionary analysis suggested that these sites were under positive selection during human evolution. 25628020 7 46 theme innovative 1389:1398 arg1 modifications 1408:1420 innovative genetic modifications 1389:1420 innovative genetic modifications for beneficial phenotypes acquired in the human lineage 1389:1476 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 1 47 theme protein 114:120 arg1 glycosylation 122:134 N-linked protein glycosylation 105:134 N-linked protein glycosylation 105:134 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 6 48 theme significant 1204:1214 arg1 decrease 1216:1223 a significant decrease 1202:1223 a significant decrease in T4 production 1202:1240 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 3 49 theme last 619:622 arg1 ancestor 631:638 the last common ancestor 615:638 the last common ancestor of Euarchonta (primates and treeshrews) 615:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 4 50 theme differentiation 782:796 arg1 cluster 771:777 cluster 771:777 cluster of differentiation 166 (CD166/ALCAM) 771:814 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 5 51 theme evolutionary 876:887 arg1 analysis 889:896 Molecular evolutionary analysis 866:896 Molecular evolutionary analysis 866:896 Molecular evolutionary analysis suggested that these sites were under positive selection during human evolution. 25628020 6 52 from site 1171:1174 arg1 removal 1132:1138 the removal 1128:1138 the removal of the glycan moiety from this site 1128:1174 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 2 53 contain have 321:324 arg1 acquisition 273:283 The acquisition 269:283 The acquisition of a novel N-glycosylation site 269:315 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 2 53 contain have 321:324 arg2 effect 338:343 significant effect 326:343 significant effect 326:343 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 5 54 theme human 962:966 arg1 evolution 968:976 human evolution 962:976 human evolution 962:976 Molecular evolutionary analysis suggested that these sites were under positive selection during human evolution. 25628020 0 55 gly N-glycosylation 25:39 arg2 sites 41:45 novel N-glycosylation sites 19:45 novel N-glycosylation sites in conserved proteins 19:67 The acquisition of novel N-glycosylation sites in conserved proteins during human evolution. 25628020 3 56 dep set 463:465 arg1 sites 490:494 2,534 N-glycosylation sites 468:494 2,534 N-glycosylation sites in 1,027 proteins 468:512 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 3 57 theme N-glycosylation 540:554 arg1 sites 556:560 112 novel N-glycosylation sites 530:560 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews) 530:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 6 58 from decrease 1216:1223 arg1 production 1231:1240 T4 production 1228:1240 T4 production 1228:1240 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 5 59 theme Molecular 866:874 arg1 analysis 889:896 Molecular evolutionary analysis 866:896 Molecular evolutionary analysis 866:896 Molecular evolutionary analysis suggested that these sites were under positive selection during human evolution. 25628020 6 60 theme moiety 1154:1159 arg1 removal 1132:1138 the removal 1128:1138 the removal of the glycan moiety from this site 1128:1174 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 4 61 theme adipocyte 707:715 arg1 protein 744:750 adipocyte plasma membrane-associated protein 707:750 adipocyte plasma membrane-associated protein (APMAP) 707:758 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 61 theme adipocyte 707:715 arg1 APMAP 753:757 APMAP 753:757 APMAP 753:757 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 2 62 theme significant 326:336 arg1 effect 338:343 significant effect 326:343 significant effect 326:343 The acquisition of a novel N-glycosylation site may have significant effect on protein structure and function, and therefore, on the phenotype. 25628020 3 63 theme Euarchonta 643:652 arg1 ancestor 631:638 the last common ancestor 615:638 the last common ancestor of Euarchonta (primates and treeshrews) 615:678 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 6 64 theme glycan 1147:1152 arg1 moiety 1154:1159 the glycan moiety 1143:1159 the glycan moiety 1143:1159 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 3 65 theme data 458:461 arg1 set 463:465 the human glycoproteome data set 434:465 the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) 434:513 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 1 66 theme important 145:153 arg1 role 155:158 an important role 142:158 an important role 142:158 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 3 67 theme 2,534 468:472 arg1 sites 490:494 2,534 N-glycosylation sites 468:494 2,534 N-glycosylation sites in 1,027 proteins 468:512 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 1 68 theme cell 240:243 arg1 folding 211:217 protein folding 203:217 protein folding 203:217 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 68 theme cell 240:243 arg1 adhesion 245:252 cell adhesion 240:252 cell adhesion 240:252 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 68 theme cell 240:243 arg1 processes 182:190 various biological processes 163:190 various biological processes 163:190 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 68 theme cell 240:243 arg1 trafficking 223:233 trafficking 223:233 trafficking 223:233 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 1 69 link N-linked 105:112 arg1 glycosylation 122:134 N-linked protein glycosylation 105:134 N-linked protein glycosylation 105:134 BACKGROUND: N-linked protein glycosylation plays an important role in various biological processes, including protein folding and trafficking, and cell adhesion and signaling. 25628020 3 70 theme human 595:599 arg1 lineage 601:607 the human lineage 591:607 the human lineage 591:607 RESULTS: We analyzed the human glycoproteome data set (2,534 N-glycosylation sites in 1,027 proteins) and identified 112 novel N-glycosylation sites in 91 proteins that arose in the human lineage since the last common ancestor of Euarchonta (primates and treeshrews). 25628020 6 71 theme hormones 1073:1080 arg1 production 1051:1060 the increased production 1037:1060 the increased production of thyroid hormones in humans, especially thyroxine (T4) 1037:1117 Notably, the Asn-76 of thyroglobulin might be involved in the increased production of thyroid hormones in humans, especially thyroxine (T4), because the removal of the glycan moiety from this site was reported to result in a significant decrease in T4 production. 25628020 7 72 theme useful 1335:1340 arg1 candidates 1342:1351 useful candidates 1335:1351 useful candidates for functional analyses 1335:1375 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 7 72 theme useful 1335:1340 arg1 sites 1298:1302 the novel N-glycosylation sites 1272:1302 the novel N-glycosylation sites described in this study 1272:1326 CONCLUSIONS: We propose that the novel N-glycosylation sites described in this study may be useful candidates for functional analyses to identify innovative genetic modifications for beneficial phenotypes acquired in the human lineage. 25628020 4 73 from Asn-196 696:702 arg1 Asn-91 761:766 Asn-91 761:766 Asn-91 in cluster of differentiation 166 (CD166/ALCAM) 761:814 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 73 from Asn-196 696:702 arg1 protein 744:750 adipocyte plasma membrane-associated protein 707:750 adipocyte plasma membrane-associated protein (APMAP) 707:758 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 73 from Asn-196 696:702 arg1 APMAP 753:757 APMAP 753:757 APMAP 753:757 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 25628020 4 73 from Asn-196 696:702 arg1 Asn-76 821:826 Asn-76 821:826 Asn-76 in thyroglobulin 821:843 Three of them, Asn-196 in adipocyte plasma membrane-associated protein (APMAP), Asn-91 in cluster of differentiation 166 (CD166/ALCAM), and Asn-76 in thyroglobulin, are human-specific. 27933952 0 0 theme Oral 107:110 arg1 Delivery 112:119 Oral Delivery 107:119 Oral Delivery 107:119 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 7 1 theme heparin 1135:1141 arg1 development 1143:1153 oral heparin development 1130:1153 oral heparin development 1130:1153 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 3 2 theme alkaline 482:489 arg1 depolymerization 491:506 alkaline depolymerization 482:506 alkaline depolymerization 482:506 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 6 3 from thrombosis 957:966 arg1 models 1001:1006 animal models 994:1006 animal models 994:1006 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 1 4 theme thromboembolic 240:253 arg1 disease 255:261 thromboembolic disease 240:261 thromboembolic disease 240:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 4 5 theme glycosylation 678:690 arg1 reaction 692:699 nonenzymatic glycosylation reaction 665:699 nonenzymatic glycosylation reaction 665:699 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 1 6 theme disease 255:261 arg1 prevention 226:235 the prevention 222:235 the prevention of thromboembolic disease 222:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 6 theme disease 255:261 arg1 treatment 209:217 the treatment 205:217 the treatment 205:217 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 6 7 theme therapeutic 925:935 arg1 effect 937:942 therapeutic effect 925:942 therapeutic effect 925:942 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 7 8 theme oral 1130:1133 arg1 development 1143:1153 oral heparin development 1130:1153 oral heparin development 1130:1153 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 6 9 theme animal 994:999 arg1 models 1001:1006 animal models 994:1006 animal models 994:1006 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 7 10 theme drug 1111:1114 arg1 candidate 1116:1124 a drug candidate 1109:1124 a drug candidate for oral heparin development 1109:1153 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 7 10 theme drug 1111:1114 arg1 EnoxaTD 1092:1098 EnoxaTD 1092:1098 EnoxaTD 1092:1098 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 3 11 contain has 404:406 arg1 enoxaparin 386:395 enoxaparin 386:395 enoxaparin 386:395 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 3 11 contain has 404:406 arg2 moiety 425:430 a reducing sugar moiety 408:430 a reducing sugar moiety 408:430 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 3 12 theme reducing 410:417 arg1 moiety 425:430 a reducing sugar moiety 408:430 a reducing sugar moiety 408:430 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 5 13 theme polysaccharide 753:766 arg1 development 773:783 polysaccharide drug development 753:783 polysaccharide drug development 753:783 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 4 14 from end-site 638:645 arg1 tetraoligomer 581:593 the tetraoligomer 577:593 the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin 577:659 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 4 15 theme enoxaparin 650:659 arg1 end-site 638:645 the end-site 634:645 the end-site of enoxaparin 634:659 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 4 15 theme enoxaparin 650:659 arg1 enoxaparin 650:659 enoxaparin 650:659 enoxaparin 650:659 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 0 16 theme End-Site-Specific 0:16 arg1 Conjugation 18:28 End-Site-Specific Conjugation 0:28 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid 0:68 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 4 17 theme LMWHs 556:560 arg1 activity 544:551 this end-site-specific activity 521:551 this end-site-specific activity of LMWHs 521:560 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 6 18 theme vein 952:955 arg1 DVT 969:971 DVT 969:971 DVT 969:971 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 18 theme vein 952:955 arg1 thrombosis 957:966 deep vein thrombosis 947:966 deep vein thrombosis (DVT) without bleeding in animal models 947:1006 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 19 theme active 869:874 arg1 enoxaparin 876:885 orally active enoxaparin 862:885 orally active enoxaparin 862:885 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 3 20 theme sugar 419:423 arg1 moiety 425:430 a reducing sugar moiety 408:430 a reducing sugar moiety 408:430 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 0 21 theme Enoxaparin 33:42 arg1 Conjugation 18:28 End-Site-Specific Conjugation 0:28 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid 0:68 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 1 22 theme choice 194:199 arg1 drug 186:189 the drug 182:189 the drug of choice for the treatment or the prevention of thromboembolic disease 182:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 22 theme choice 194:199 arg1 heparins 155:162 low molecular weight heparins 134:162 low molecular weight heparins (LMWHs) 134:170 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 22 theme choice 194:199 arg1 Heparin 122:128 Heparin 122:128 Heparin 122:128 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 3 23 theme polysaccharide 451:464 arg1 end-site 439:446 the end-site 435:446 the end-site of polysaccharide 435:464 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 3 23 theme polysaccharide 451:464 arg1 polysaccharide 451:464 polysaccharide 451:464 polysaccharide 451:464 In particular, enoxaparin, which has a reducing sugar moiety at the end-site of polysaccharide, is prepared by alkaline depolymerization. 27933952 0 24 theme Tetradeoxycholic 48:63 arg1 Acid 65:68 Tetradeoxycholic Acid 48:68 Tetradeoxycholic Acid 48:68 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 5 25 theme end-site-specific 706:722 arg1 important 739:747 important 739:747 important 739:747 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 5 25 theme end-site-specific 706:722 arg1 conjugation 724:734 The end-site-specific conjugation 702:734 The end-site-specific conjugation 702:734 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 6 26 theme tetraDOCA 891:899 arg1 EnoxaTD 912:918 EnoxaTD 912:918 EnoxaTD 912:918 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 26 theme tetraDOCA 891:899 arg1 conjugate 901:909 tetraDOCA conjugate 891:909 tetraDOCA conjugate (EnoxaTD) 891:919 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 27 contain had 921:923 arg2 effect 937:942 therapeutic effect 925:942 therapeutic effect 925:942 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 27 contain had 921:923 arg1 EnoxaTD 912:918 EnoxaTD 912:918 EnoxaTD 912:918 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 27 contain had 921:923 arg1 enoxaparin 876:885 orally active enoxaparin 862:885 orally active enoxaparin 862:885 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 27 contain had 921:923 arg1 conjugate 901:909 tetraDOCA conjugate 891:909 tetraDOCA conjugate (EnoxaTD) 891:919 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 5 28 theme polysaccharides 817:831 arg1 heterogeneity 800:812 the heterogeneity 796:812 the heterogeneity of polysaccharides 796:831 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 7 29 theme end-specific 1039:1050 arg1 conjugation 1052:1062 end-specific conjugation 1039:1062 end-specific conjugation 1039:1062 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 1 30 theme low 134:136 arg1 drug 186:189 the drug 182:189 the drug of choice for the treatment or the prevention of thromboembolic disease 182:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 30 theme low 134:136 arg1 LMWHs 165:169 LMWHs 165:169 LMWHs 165:169 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 30 theme low 134:136 arg1 heparins 155:162 low molecular weight heparins 134:162 low molecular weight heparins (LMWHs) 134:170 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 30 theme low 134:136 arg1 Heparin 122:128 Heparin 122:128 Heparin 122:128 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 0 31 theme Acid 65:68 arg1 Conjugation 18:28 End-Site-Specific Conjugation 0:28 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid 0:68 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 4 32 dep TetraDOCA 616:624 arg1 TD 627:628 TD 627:628 TD 627:628 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 1 33 theme molecular 138:146 arg1 drug 186:189 the drug 182:189 the drug of choice for the treatment or the prevention of thromboembolic disease 182:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 33 theme molecular 138:146 arg1 LMWHs 165:169 LMWHs 165:169 LMWHs 165:169 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 33 theme molecular 138:146 arg1 heparins 155:162 low molecular weight heparins 134:162 low molecular weight heparins (LMWHs) 134:170 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 33 theme molecular 138:146 arg1 Heparin 122:128 Heparin 122:128 Heparin 122:128 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 4 34 theme end-site-specific 526:542 arg1 activity 544:551 this end-site-specific activity 521:551 this end-site-specific activity of LMWHs 521:560 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 7 35 theme conjugation 1052:1062 arg1 importance 1025:1034 the importance 1021:1034 the importance of end-specific conjugation 1021:1062 Considering the importance of end-specific conjugation, these results suggest that EnoxaTD could be a drug candidate for oral heparin development. 27933952 5 36 gly heterogeneity 800:812 arg1 polysaccharides 817:831 polysaccharides 817:831 polysaccharides 817:831 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 2 37 theme Different 264:272 arg1 methods 274:280 Different methods 264:280 Different methods 264:280 Different methods are employed to prepare the LMWHs that are clinically approved for the market currently. 27933952 1 38 theme weight 148:153 arg1 drug 186:189 the drug 182:189 the drug of choice for the treatment or the prevention of thromboembolic disease 182:261 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 38 theme weight 148:153 arg1 LMWHs 165:169 LMWHs 165:169 LMWHs 165:169 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 38 theme weight 148:153 arg1 heparins 155:162 low molecular weight heparins 134:162 low molecular weight heparins (LMWHs) 134:170 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 1 38 theme weight 148:153 arg1 Heparin 122:128 Heparin 122:128 Heparin 122:128 Heparin and low molecular weight heparins (LMWHs) have been the drug of choice for the treatment or the prevention of thromboembolic disease. 27933952 4 39 theme acid 610:613 arg1 tetraoligomer 581:593 the tetraoligomer 577:593 the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin 577:659 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 4 40 theme deoxycholic 598:608 arg1 acid 610:613 deoxycholic acid 598:613 deoxycholic acid (TetraDOCA; TD) 598:629 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 4 40 theme deoxycholic 598:608 arg1 TetraDOCA 616:624 TetraDOCA 616:624 TetraDOCA 616:624 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 5 41 theme drug 768:771 arg1 development 773:783 polysaccharide drug development 753:783 polysaccharide drug development 753:783 The end-site-specific conjugation is important for polysaccharide drug development because of the heterogeneity of polysaccharides. 27933952 4 42 theme nonenzymatic 665:676 arg1 reaction 692:699 nonenzymatic glycosylation reaction 665:699 nonenzymatic glycosylation reaction 665:699 Focusing on this end-site-specific activity of LMWHs, we conjugated the tetraoligomer of deoxycholic acid (TetraDOCA; TD) at the end-site of enoxaparin via nonenzymatic glycosylation reaction. 27933952 0 43 theme Nonenzymatic 76:87 arg1 Glycosylation 89:101 Nonenzymatic Glycosylation 76:101 Nonenzymatic Glycosylation 76:101 End-Site-Specific Conjugation of Enoxaparin and Tetradeoxycholic Acid Using Nonenzymatic Glycosylation for Oral Delivery. 27933952 6 44 theme deep 947:950 arg1 DVT 969:971 DVT 969:971 DVT 969:971 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 27933952 6 44 theme deep 947:950 arg1 thrombosis 957:966 deep vein thrombosis 947:966 deep vein thrombosis (DVT) without bleeding in animal models 947:1006 This study also showed that orally active enoxaparin and tetraDOCA conjugate (EnoxaTD) had therapeutic effect on deep vein thrombosis (DVT) without bleeding in animal models. 26554003 4 0 theme ± 934:934 arg1 residues 938:945 ± 4 residues 934:945 ± 4 residues that could represent O-glycosites 934:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 3 1 theme particular 776:785 arg1 ADAM17 787:792 particular ADAM17 776:792 particular ADAM17 776:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 6 2 dep ex 1298:1299 arg1 vivo 1301:1304 vivo 1301:1304 vivo 1301:1304 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 0 3 theme site-specific 73:85 arg1 O-glycosylation 87:101 site-specific O-glycosylation 73:101 site-specific O-glycosylation 73:101 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 6 4 theme in 1334:1335 arg1 isoform 1285:1291 the GalNAc-T2 isoform 1271:1291 the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts 1271:1366 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 4 5 gly O-glycosites 968:979 arg2 O-glycosites 968:979 O-glycosites 968:979 O-glycosites 968:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 5 6 theme 12 1081:1082 arg1 shedding 1060:1067 shedding 1060:1067 shedding of at least 12 of these proteins 1060:1100 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 5 7 theme ADAM 1019:1022 arg1 assays 1033:1038 in vitro GalNAc-T enzyme and ADAM cleavage assays 990:1038 assays 1033:1038 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 1 8 theme cell 267:270 arg1 signaling 272:280 cell signaling 267:280 cell signaling 267:280 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 7 9 theme ectodomain 1461:1470 arg1 shedding 1472:1479 ectodomain shedding 1461:1479 ectodomain shedding 1461:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 5 10 theme cleavage 1024:1031 arg1 assays 1033:1038 in vitro GalNAc-T enzyme and ADAM cleavage assays 990:1038 assays 1033:1038 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 3 11 theme distinct 593:600 arg1 isoforms 644:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 2 12 theme juxtamembrane 341:353 arg1 region 355:360 the immediate extracellular juxtamembrane region 313:360 the immediate extracellular juxtamembrane region 313:360 Ectodomain shedding occurs in the immediate extracellular juxtamembrane region, which is also where O-glycosylation is often found and examples of crosstalk between shedding and O-glycosylation have been reported. 26554003 5 13 theme GalNAc-T 999:1006 arg1 enzyme 1008:1013 in vitro GalNAc-T enzyme and ADAM cleavage assays 990:1038 enzyme 1008:1013 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 2 14 theme Ectodomain 283:292 arg1 shedding 294:301 Ectodomain shedding 283:301 Ectodomain shedding 283:301 Ectodomain shedding occurs in the immediate extracellular juxtamembrane region, which is also where O-glycosylation is often found and examples of crosstalk between shedding and O-glycosylation have been reported. 26554003 4 15 dep shedding 861:868 arg1 included 901:908 included 901:908 included Ser/Thr residues within ± 4 residues that could represent O-glycosites 901:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 6 16 dep in 1334:1335 arg1 vivo 1337:1340 vivo 1337:1340 vivo 1337:1340 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 6 17 theme isogenic 1309:1316 arg1 models 1323:1328 isogenic cell models 1309:1328 isogenic cell models 1309:1328 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 0 18 theme systematic 2:11 arg1 study 13:17 A systematic study 0:17 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation 0:101 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 3 19 theme site-specific 551:563 arg1 O-glycosylation 565:579 site-specific O-glycosylation 551:579 site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 551:651 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 2 20 theme immediate 317:325 arg1 region 355:360 the immediate extracellular juxtamembrane region 313:360 the immediate extracellular juxtamembrane region 313:360 Ectodomain shedding occurs in the immediate extracellular juxtamembrane region, which is also where O-glycosylation is often found and examples of crosstalk between shedding and O-glycosylation have been reported. 26554003 2 21 theme extracellular 327:339 arg1 region 355:360 the immediate extracellular juxtamembrane region 313:360 the immediate extracellular juxtamembrane region 313:360 Ectodomain shedding occurs in the immediate extracellular juxtamembrane region, which is also where O-glycosylation is often found and examples of crosstalk between shedding and O-glycosylation have been reported. 26554003 6 22 theme Galnt2 1351:1356 arg1 knockouts 1358:1366 mouse Galnt2 knockouts 1345:1366 mouse Galnt2 knockouts 1345:1366 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 1 23 theme common 185:190 arg1 process 192:198 a common process 183:198 a common process that releases the extracellular domain from the cell and activates cell signaling 183:280 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 1 23 theme common 185:190 arg1 shedding 114:121 Regulated shedding 104:121 Regulated shedding of the ectodomain of cell membrane proteins by proteases 104:178 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 6 24 theme cell 1318:1321 arg1 models 1323:1328 isogenic cell models 1309:1328 isogenic cell models 1309:1328 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 4 25 theme Ser/Thr 910:916 arg1 residues 918:925 Ser/Thr residues 910:925 Ser/Thr residues within ± 4 residues that could represent O-glycosites 910:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 3 26 theme GalNAc-T 634:641 arg1 isoforms 644:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 7 27 theme wider 1414:1418 arg1 role 1420:1423 a wider role 1412:1423 a wider role of site-specific O-glycosylation in ectodomain shedding 1412:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 7 28 theme compelling 1388:1397 arg1 evidence 1399:1406 compelling evidence 1388:1406 compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding 1388:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 5 29 theme in 990:991 arg1 enzyme 1008:1013 in vitro GalNAc-T enzyme and ADAM cleavage assays 990:1038 enzyme 1008:1013 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 3 30 theme polypeptide 602:612 arg1 isoforms 644:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 7 31 from role 1420:1423 arg1 shedding 1472:1479 ectodomain shedding 1461:1479 ectodomain shedding 1461:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 1 32 theme Regulated 104:112 arg1 process 192:198 a common process 183:198 a common process that releases the extracellular domain from the cell and activates cell signaling 183:280 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 1 32 theme Regulated 104:112 arg1 shedding 114:121 Regulated shedding 104:121 Regulated shedding of the ectodomain of cell membrane proteins by proteases 104:178 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 3 33 theme O-glycosylation 565:579 arg1 potential 538:546 the potential 534:546 the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17 534:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 3 34 theme Disintegrin 705:715 arg1 subfamily 746:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily 699:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17 699:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 3 35 theme GalNAc-transferase 614:631 arg1 isoforms 644:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms 593:651 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 2 36 theme crosstalk 430:438 arg1 examples 418:425 examples 418:425 examples of crosstalk between shedding and O-glycosylation 418:475 Ectodomain shedding occurs in the immediate extracellular juxtamembrane region, which is also where O-glycosylation is often found and examples of crosstalk between shedding and O-glycosylation have been reported. 26554003 3 37 from subfamily 746:754 arg1 ADAM17 787:792 particular ADAM17 776:792 particular ADAM17 776:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 6 38 theme ex 1298:1299 arg1 isoform 1285:1291 the GalNAc-T2 isoform 1271:1291 the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts 1271:1366 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 0 39 theme modulation 22:31 arg1 study 13:17 A systematic study 0:17 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation 0:101 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 7 40 theme O-glycosylation 1442:1456 arg1 role 1420:1423 a wider role 1412:1423 a wider role of site-specific O-glycosylation in ectodomain shedding 1412:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 7 41 theme site-specific 1428:1440 arg1 O-glycosylation 1442:1456 site-specific O-glycosylation 1428:1456 site-specific O-glycosylation 1428:1456 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 1 42 theme extracellular 218:230 arg1 domain 232:237 the extracellular domain 214:237 the extracellular domain from the cell 214:251 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 0 43 theme ectodomain 50:59 arg1 shedding 61:68 ADAM-mediated ectodomain shedding 36:68 ADAM-mediated ectodomain shedding 36:68 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 7 44 gly O-glycosylation 1442:1456 arg1 ectodomain 1461:1470 ectodomain shedding 1461:1479 ectodomain shedding 1461:1479 The study provides compelling evidence for a wider role of site-specific O-glycosylation in ectodomain shedding. 26554003 4 45 theme processing 884:893 arg1 sites 895:899 the processing sites 880:899 the processing sites 880:899 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 0 46 theme ADAM-mediated 36:48 arg1 shedding 61:68 ADAM-mediated ectodomain shedding 36:68 ADAM-mediated ectodomain shedding 36:68 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 1 47 from cell 248:251 arg1 domain 232:237 the extracellular domain 214:237 the extracellular domain from the cell 214:251 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 3 48 theme ADAM 740:743 arg1 subfamily 746:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily 699:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17 699:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 6 49 theme GalNAc-T2 1275:1283 arg1 isoform 1285:1291 the GalNAc-T2 isoform 1271:1291 the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts 1271:1366 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 1 50 theme ectodomain 130:139 arg1 process 192:198 a common process 183:198 a common process that releases the extracellular domain from the cell and activates cell signaling 183:280 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 1 50 theme ectodomain 130:139 arg1 shedding 114:121 Regulated shedding 104:121 Regulated shedding of the ectodomain of cell membrane proteins by proteases 104:178 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 6 51 theme mouse 1345:1349 arg1 knockouts 1358:1366 mouse Galnt2 knockouts 1345:1366 mouse Galnt2 knockouts 1345:1366 Using TNF-α as an example, we confirmed that shedding mediated by ADAM17 is coregulated by O-glycosylation controlled by the GalNAc-T2 isoform both ex vivo in isogenic cell models and in vivo in mouse Galnt2 knockouts. 26554003 5 52 theme proteins 1093:1100 arg1 12 1081:1082 12 1081:1082 12 1081:1082 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 5 52 theme proteins 1093:1100 arg1 proteins 1093:1100 these proteins 1087:1100 these proteins 1087:1100 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 4 53 theme membrane 810:817 arg1 proteins 819:826 25 membrane proteins 807:826 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites 807:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 3 54 theme Metalloproteinase 721:737 arg1 ADAM 740:743 Metalloproteinase (ADAM) 721:744 Metalloproteinase (ADAM) 721:744 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 5 55 used used 985:988 arg2 We 982:983 We 982:983 We 982:983 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 1 56 theme cell 144:147 arg1 proteins 158:165 cell membrane proteins 144:165 cell membrane proteins 144:165 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 5 57 dep in 990:991 arg1 vitro 993:997 vitro 993:997 vitro 993:997 We used in vitro GalNAc-T enzyme and ADAM cleavage assays to demonstrate that shedding of at least 12 of these proteins are potentially coregulated by O-glycosylation. 26554003 3 58 theme ectodomain 667:676 arg1 shedding 678:685 ectodomain shedding 667:685 ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17 667:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 1 59 theme membrane 149:156 arg1 proteins 158:165 cell membrane proteins 144:165 cell membrane proteins 144:165 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 0 60 theme shedding 61:68 arg1 modulation 22:31 modulation 22:31 modulation of ADAM-mediated ectodomain shedding 22:68 A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. 26554003 3 61 theme proteases 759:767 arg1 subfamily 746:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily 699:754 the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17 699:792 Here, we systematically investigated the potential of site-specific O-glycosylation mediated by distinct polypeptide GalNAc-transferase (GalNAc-T) isoforms to coregulate ectodomain shedding mediated by the A Disintegrin And Metalloproteinase (ADAM) subfamily of proteases and in particular ADAM17. 26554003 4 62 theme ADAM17 854:859 arg1 shedding 861:868 ADAM17 shedding 854:868 ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites 854:979 We analyzed 25 membrane proteins that are known to undergo ADAM17 shedding and where the processing sites included Ser/Thr residues within ± 4 residues that could represent O-glycosites. 26554003 1 63 theme proteins 158:165 arg1 proteins 158:165 cell membrane proteins 144:165 cell membrane proteins 144:165 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 26554003 1 63 theme proteins 158:165 arg1 ectodomain 130:139 the ectodomain 126:139 the ectodomain of cell membrane proteins 126:165 Regulated shedding of the ectodomain of cell membrane proteins by proteases is a common process that releases the extracellular domain from the cell and activates cell signaling. 29190644 1 0 gly glycoprotein 185:196 arg1 butyrylcholinesterase 151:171 Human butyrylcholinesterase 145:171 Human butyrylcholinesterase (BChE) 145:178 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 1 0 gly glycoprotein 185:196 arg1 glycoprotein 185:196 a glycoprotein 183:196 a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents 183:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 3 1 from profile 440:446 arg1 systems 457:463 these systems 451:463 these systems 451:463 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 2 2 theme non-human 356:364 arg1 animals 404:410 animals 404:410 animals 404:410 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 2 theme non-human 356:364 arg1 plants 394:399 plants 394:399 plants 394:399 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 2 theme non-human 356:364 arg1 systems 377:383 non-human expression systems 356:383 non-human expression systems 356:383 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 5 3 gly glycosylation 840:852 arg2 site 854:857 populating glycosylation site ASN241 829:864 populating glycosylation site ASN241 829:864 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 5 3 gly glycosylation 840:852 arg2 ASN241 859:864 ASN241 859:864 ASN241 859:864 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 5 4 theme BChE 883:886 arg1 glycoforms 888:897 monomeric human BChE glycoforms 867:897 monomeric human BChE glycoforms 867:897 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 3 5 theme glycosylation 507:519 arg1 profile 521:527 the human glycosylation profile 497:527 the human glycosylation profile 497:527 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 1 6 theme nerve 269:273 arg1 agents 275:280 organophosphorus (OP) nerve agents 247:280 organophosphorus (OP) nerve agents 247:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 6 7 dep structure 991:999 arg1 the 987:989 the 987:989 the 987:989 Our simulations indicate that the structure and function of human BChE are significantly affected by the absence of glycan 241. 29190644 3 8 from profile 521:527 arg1 profile 440:446 the glycosylation profile 422:446 the glycosylation profile in these systems 422:463 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 3 8 from profile 521:527 arg1 different 482:490 different 482:490 different 482:490 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 4 9 theme BChE 744:747 arg1 domain 765:770 the BChE tetramerization domain 740:770 the BChE tetramerization domain 740:770 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 2 10 from BChE 348:351 arg1 animals 404:410 animals 404:410 animals 404:410 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 10 from BChE 348:351 arg1 plants 394:399 plants 394:399 plants 394:399 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 10 from BChE 348:351 arg1 systems 377:383 non-human expression systems 356:383 non-human expression systems 356:383 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 3 11 theme human 501:505 arg1 profile 521:527 the human glycosylation profile 497:527 the human glycosylation profile 497:527 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 5 12 theme glycosylation 840:852 arg1 site 854:857 populating glycosylation site ASN241 829:864 populating glycosylation site ASN241 829:864 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 0 13 theme site 132:135 arg1 importance 104:113 The importance 100:113 The importance of glycosylation site ASN241 100:142 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 6 14 theme glycan 1073:1078 arg1 absence 1062:1068 the absence 1058:1068 the absence of glycan 241 1058:1082 Our simulations indicate that the structure and function of human BChE are significantly affected by the absence of glycan 241. 29190644 2 15 theme commercial 287:296 arg1 production 298:307 commercial production 287:307 commercial production of BChE 287:315 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 3 16 theme glycosylation 426:438 arg1 profile 440:446 the glycosylation profile 422:446 the glycosylation profile in these systems 422:463 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 3 16 theme glycosylation 426:438 arg1 different 482:490 different 482:490 different 482:490 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 0 17 theme glycosylation 118:130 arg1 site 132:135 glycosylation site ASN241 118:142 glycosylation site ASN241 118:142 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 4 18 theme tetramerization 749:763 arg1 domain 765:770 the BChE tetramerization domain 740:770 the BChE tetramerization domain 740:770 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 5 19 gly glycosylated 943:954 arg1 glycoforms 888:897 monomeric human BChE glycoforms 867:897 monomeric human BChE glycoforms 867:897 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 4 20 theme close 721:725 arg1 proximity 727:735 its close proximity 717:735 its close proximity to the BChE tetramerization domain and the active site gorge 717:796 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 1 21 theme capable 198:204 arg1 butyrylcholinesterase 151:171 Human butyrylcholinesterase 145:171 Human butyrylcholinesterase (BChE) 145:178 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 1 21 theme capable 198:204 arg1 glycoprotein 185:196 a glycoprotein 183:196 a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents 183:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 0 22 theme Structural 0:9 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of human glycoprotein butyrylcholinesterase 0:62 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 0 23 theme human 23:27 arg1 butyrylcholinesterase 42:62 human glycoprotein butyrylcholinesterase 23:62 human glycoprotein butyrylcholinesterase 23:62 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 0 24 gly glycosylation 118:130 arg2 site 132:135 glycosylation site ASN241 118:142 glycosylation site ASN241 118:142 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 0 24 gly glycosylation 118:130 arg2 ASN241 137:142 ASN241 137:142 ASN241 137:142 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 2 25 theme BChE 312:315 arg1 production 298:307 commercial production 287:307 commercial production of BChE 287:315 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 1 26 theme toxic 223:227 arg1 agents 275:280 organophosphorus (OP) nerve agents 247:280 organophosphorus (OP) nerve agents 247:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 1 26 theme toxic 223:227 arg1 compounds 229:237 toxic compounds 223:237 toxic compounds such as organophosphorus (OP) nerve agents 223:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 0 27 theme butyrylcholinesterase 42:62 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of human glycoprotein butyrylcholinesterase 0:62 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 4 28 attach attached 643:650 arg2 important 700:708 important 700:708 important 700:708 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 4 28 attach attached 643:650 arg2 glycan 636:641 the glycan 632:641 the glycan attached to ASN241 632:660 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 4 28 attach attached 643:650 arg1 ASN241 655:660 ASN241 655:660 ASN241 655:660 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 0 29 theme glycoprotein 29:40 arg1 butyrylcholinesterase 42:62 human glycoprotein butyrylcholinesterase 23:62 human glycoprotein butyrylcholinesterase 23:62 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 0 30 dep using 64:68 arg1 importance 104:113 The importance 100:113 The importance of glycosylation site ASN241 100:142 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 5 31 theme populating 829:838 arg1 site 854:857 populating glycosylation site ASN241 829:864 populating glycosylation site ASN241 829:864 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 0 32 theme atomistic 70:78 arg1 dynamics 90:97 atomistic molecular dynamics 70:97 atomistic molecular dynamics 70:97 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 3 33 from changes 552:558 arg1 function 584:591 function 584:591 function 584:591 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 3 33 from changes 552:558 arg1 structure 570:578 BChE's structure 563:578 BChE's structure 563:578 However, the glycosylation profile in these systems is significantly different from the human glycosylation profile, which could result in changes in BChE's structure and function. 29190644 5 34 gly glycoforms 888:897 arg1 BChE 883:886 monomeric human BChE glycoforms 867:897 monomeric human BChE glycoforms 867:897 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 0 35 gly glycoprotein 29:40 arg1 glycoprotein 29:40 human glycoprotein butyrylcholinesterase 23:62 human glycoprotein butyrylcholinesterase 23:62 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 6 36 theme human 1017:1021 arg1 BChE 1023:1026 human BChE 1017:1026 human BChE 1017:1026 Our simulations indicate that the structure and function of human BChE are significantly affected by the absence of glycan 241. 29190644 1 37 theme Human 145:149 arg1 BChE 174:177 BChE 174:177 BChE 174:177 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 1 37 theme Human 145:149 arg1 butyrylcholinesterase 151:171 Human butyrylcholinesterase 145:171 Human butyrylcholinesterase (BChE) 145:178 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 1 37 theme Human 145:149 arg1 glycoprotein 185:196 a glycoprotein 183:196 a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents 183:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 6 38 theme BChE 1023:1026 arg1 structure 991:999 structure 991:999 structure 991:999 Our simulations indicate that the structure and function of human BChE are significantly affected by the absence of glycan 241. 29190644 6 38 theme BChE 1023:1026 arg1 function 1005:1012 function 1005:1012 function 1005:1012 Our simulations indicate that the structure and function of human BChE are significantly affected by the absence of glycan 241. 29190644 5 39 theme human 877:881 arg1 glycoforms 888:897 monomeric human BChE glycoforms 867:897 monomeric human BChE glycoforms 867:897 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 1 40 theme organophosphorus 247:262 arg1 agents 275:280 organophosphorus (OP) nerve agents 247:280 organophosphorus (OP) nerve agents 247:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 29190644 4 41 theme site 787:790 arg1 gorge 792:796 the active site gorge 776:796 the active site gorge 776:796 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 5 42 theme monomeric 867:875 arg1 glycoforms 888:897 monomeric human BChE glycoforms 867:897 monomeric human BChE glycoforms 867:897 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 5 43 theme site 854:857 arg1 effects 818:824 the effects 814:824 the effects of populating glycosylation site ASN241 814:864 To investigate the effects of populating glycosylation site ASN241, monomeric human BChE glycoforms were simulated with and without site ASN241 glycosylated. 29190644 0 44 theme molecular 80:88 arg1 dynamics 90:97 atomistic molecular dynamics 70:97 atomistic molecular dynamics 70:97 Structural analysis of human glycoprotein butyrylcholinesterase using atomistic molecular dynamics: The importance of glycosylation site ASN241. 29190644 4 45 theme active 780:785 arg1 gorge 792:796 the active site gorge 776:796 the active site gorge 776:796 From our investigation, we found that the glycan attached to ASN241 is both structurally and functionally important due to its close proximity to the BChE tetramerization domain and the active site gorge. 29190644 2 46 theme expression 366:375 arg1 animals 404:410 animals 404:410 animals 404:410 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 46 theme expression 366:375 arg1 plants 394:399 plants 394:399 plants 394:399 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 2 46 theme expression 366:375 arg1 systems 377:383 non-human expression systems 356:383 non-human expression systems 356:383 For commercial production of BChE, it is practical to synthesize BChE in non-human expression systems, such as plants or animals. 29190644 1 47 theme OP 265:266 arg1 agents 275:280 organophosphorus (OP) nerve agents 247:280 organophosphorus (OP) nerve agents 247:280 Human butyrylcholinesterase (BChE) is a glycoprotein capable of bioscavenging toxic compounds such as organophosphorus (OP) nerve agents. 28104755 12 0 gly N-glycosylation 1946:1960 arg1 SMPDL3A 2040:2046 human SMPDL3A 2034:2046 human SMPDL3A 2034:2046 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 11 1 theme predicted 1744:1752 arg1 peptide 1783:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 1 theme predicted 1744:1752 arg1 essential 1812:1820 essential 1812:1820 essential 1812:1820 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 6 2 theme Tunicamycin 910:920 arg1 treatment 927:935 Tunicamycin (TM) treatment 910:935 Tunicamycin (TM) treatment 910:935 Tunicamycin (TM) treatment resulted in expression of non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome. 28104755 10 3 theme activity 1685:1692 arg1 preservation 1659:1670 preservation 1659:1670 preservation of enzymatic activity 1659:1692 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 10 3 theme activity 1685:1692 arg1 protection 1612:1621 protection 1612:1621 protection against proteasomal degradation 1612:1653 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 1 4 link N-linked 251:258 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 4 link N-linked 251:258 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 4 link N-linked 251:258 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 5 5 gly N-glycosylation 698:712 arg1 SMPDL3A 765:771 human SMPDL3A 759:771 human SMPDL3A 759:771 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 11 6 theme signal 1776:1781 arg1 peptide 1783:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 6 theme signal 1776:1781 arg1 essential 1812:1820 essential 1812:1820 essential 1812:1820 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 7 7 theme TM-treated 1108:1117 arg1 cells 1119:1123 TM-treated cells 1108:1123 TM-treated cells 1108:1123 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 8 8 theme purified 1229:1236 arg1 SMPDL3A 1250:1256 purified recombinant SMPDL3A 1229:1256 purified recombinant SMPDL3A 1229:1256 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 10 9 theme proteasomal 1631:1641 arg1 degradation 1643:1653 proteasomal degradation 1631:1653 proteasomal degradation 1631:1653 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 8 10 theme activity 1313:1320 arg1 loss 1287:1290 significant loss 1275:1290 significant loss of phosphodiesterase activity 1275:1320 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 7 11 theme SMPDL3A 1097:1103 arg1 levels 1087:1092 levels 1087:1092 levels of SMPDL3A in TM-treated cells 1087:1123 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 8 12 theme SMPDL3A 1250:1256 arg1 deglycosylation 1210:1224 Enzymatic deglycosylation 1200:1224 Enzymatic deglycosylation of purified recombinant SMPDL3A 1200:1256 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 6 13 theme SMPDL3A 980:986 arg1 expression 949:958 expression 949:958 expression of non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome 949:1052 Tunicamycin (TM) treatment resulted in expression of non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome. 28104755 11 14 theme CHO 1906:1908 arg1 cells 1910:1914 CHO cells 1906:1914 CHO cells 1906:1914 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 5 15 theme N-glycosylation 698:712 arg1 roles 689:693 the roles 685:693 the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A 685:771 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 2 16 theme acid 306:309 arg1 sphingomyelinase 311:326 acid sphingomyelinase 306:326 acid sphingomyelinase (aSMase) 306:335 SMPDL3A is highly homologous to acid sphingomyelinase (aSMase), but unlike aSMase cannot cleave sphingomyelin. 28104755 2 16 theme acid 306:309 arg1 aSMase 329:334 aSMase 329:334 aSMase 329:334 SMPDL3A is highly homologous to acid sphingomyelinase (aSMase), but unlike aSMase cannot cleave sphingomyelin. 28104755 1 17 theme identified 201:210 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 17 theme identified 201:210 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 17 theme identified 201:210 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 5 18 theme site-directed 814:826 arg1 mutagenesis 828:838 site-directed mutagenesis 814:838 site-directed mutagenesis 814:838 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 5 19 theme THP-1 853:857 arg1 macrophages 859:869 THP-1 macrophages 853:869 THP-1 macrophages 853:869 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 8 20 theme significant 1275:1285 arg1 loss 1287:1290 significant loss 1275:1290 significant loss of phosphodiesterase activity 1275:1320 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 4 21 theme recent 474:479 arg1 studies 492:498 recent structural studies 474:498 recent structural studies 474:498 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 9 22 gly N-glycosylation 1363:1377 arg2 sites 1379:1383 individual N-glycosylation sites 1352:1383 individual N-glycosylation sites 1352:1383 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 1 23 theme Sphingomyelin 132:144 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 23 theme Sphingomyelin 132:144 arg1 SMPDL3A 178:184 SMPDL3A 178:184 SMPDL3A 178:184 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 23 theme Sphingomyelin 132:144 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 23 theme Sphingomyelin 132:144 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 9 24 theme individual 1352:1361 arg1 sites 1379:1383 individual N-glycosylation sites 1352:1383 individual N-glycosylation sites 1352:1383 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 5 25 theme mutagenesis 828:838 arg1 inhibitors 780:789 inhibitors 780:789 inhibitors of N-glycosylation and site-directed mutagenesis 780:838 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 11 26 attach removed 1860:1866 arg1 protein 1884:1890 the mature protein 1873:1890 the mature protein secreted from CHO cells 1873:1914 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 26 attach removed 1860:1866 arg2 peptide 1783:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 26 attach removed 1860:1866 arg2 essential 1812:1820 essential 1812:1820 essential 1812:1820 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 5 27 theme human 895:899 arg1 SMPDL3A 901:907 human SMPDL3A 895:907 human SMPDL3A 895:907 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 10 28 theme N-linked 1536:1543 arg1 SMPDL3A 1524:1530 SMPDL3A 1524:1530 SMPDL3A 1524:1530 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 10 28 theme N-linked 1536:1543 arg1 site 1545:1548 an N-linked site 1533:1548 an N-linked site conserved throughout the aSMase-like family 1533:1592 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 5 29 theme SMPDL3A 765:771 arg1 secretion 733:741 secretion 733:741 secretion 733:741 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 5 29 theme SMPDL3A 765:771 arg1 expression 721:730 expression 721:730 expression 721:730 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 5 29 theme SMPDL3A 765:771 arg1 activity 747:754 activity 747:754 activity 747:754 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 9 30 theme sites 1379:1383 arg1 mutagenesis 1337:1347 Site-directed mutagenesis 1323:1347 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A 1323:1394 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 12 31 theme SMPDL3A 2040:2046 arg1 stability 1997:2005 the intracellular stability 1979:2005 the intracellular stability 1979:2005 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 12 31 theme SMPDL3A 2040:2046 arg1 activity 2022:2029 activity 2022:2029 activity 2022:2029 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 12 31 theme SMPDL3A 2040:2046 arg1 secretion 2008:2016 secretion 2008:2016 secretion 2008:2016 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 4 32 theme unexpected 525:534 arg1 preferences 546:556 these unexpected substrate preferences 519:556 these unexpected substrate preferences 519:556 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 11 33 theme mature 1877:1882 arg1 protein 1884:1890 the mature protein 1873:1890 the mature protein secreted from CHO cells 1873:1914 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 1 34 gly glycoprotein 260:271 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 34 gly glycoprotein 260:271 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 34 gly glycoprotein 260:271 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 9 35 theme Asn69 1424:1428 arg1 glycosylation 1407:1419 glycosylation 1407:1419 glycosylation of Asn69 and Asn222 1407:1439 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 10 36 gly Glycosylation 1497:1509 arg1 Asn356 1514:1519 Asn356 1514:1519 Asn356 1514:1519 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 0 37 theme human 19:23 arg1 phosphodiesterase 39:55 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 9 38 theme Asn222 1434:1439 arg1 glycosylation 1407:1419 glycosylation 1407:1419 glycosylation of Asn69 and Asn222 1407:1439 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 12 39 theme intracellular 1983:1995 arg1 stability 1997:2005 the intracellular stability 1979:2005 the intracellular stability 1979:2005 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 4 40 theme SMPDL3A 581:587 arg1 biology 589:595 SMPDL3A biology 581:595 SMPDL3A biology 581:595 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 0 41 theme phosphodiesterase 39:55 arg1 N-glycosylation 0:14 N-glycosylation 0:14 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 0:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 7 42 theme phosphodiesterase 1172:1188 arg1 activity 1190:1197 phosphodiesterase activity 1172:1197 phosphodiesterase activity 1172:1197 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 11 43 theme first 1710:1714 arg1 evidence 1729:1736 the first experimental evidence 1706:1736 the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells 1706:1914 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 0 44 dep phosphodiesterase 39:55 arg1 SMPDL3A 71:77 SMPDL3A 71:77 SMPDL3A 71:77 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 0 44 dep phosphodiesterase 39:55 arg1 3A 67:68 acid-like 3A 57:68 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 4 45 theme many 559:562 arg1 aspects 570:576 many other aspects 559:576 many other aspects 559:576 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 9 46 theme N-glycans 1472:1480 arg1 secretion 1486:1494 secretion 1486:1494 secretion 1486:1494 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 9 46 theme N-glycans 1472:1480 arg1 maturation 1454:1463 maturation 1454:1463 maturation of its N-glycans 1454:1480 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 1 47 theme acid-like 164:172 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 47 theme acid-like 164:172 arg1 SMPDL3A 178:184 SMPDL3A 178:184 SMPDL3A 178:184 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 47 theme acid-like 164:172 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 47 theme acid-like 164:172 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 10 48 from Glycosylation 1497:1509 arg1 SMPDL3A 1524:1530 SMPDL3A 1524:1530 SMPDL3A 1524:1530 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 10 48 from Glycosylation 1497:1509 arg1 site 1545:1548 an N-linked site 1533:1548 an N-linked site conserved throughout the aSMase-like family 1533:1592 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 11 49 theme N-terminal 1765:1774 arg1 peptide 1783:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 49 theme N-terminal 1765:1774 arg1 essential 1812:1820 essential 1812:1820 essential 1812:1820 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 8 50 gly deglycosylation 1210:1224 arg1 SMPDL3A 1250:1256 purified recombinant SMPDL3A 1229:1256 purified recombinant SMPDL3A 1229:1256 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 1 51 theme N-linked 251:258 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 51 theme N-linked 251:258 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 51 theme N-linked 251:258 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 11 52 theme 22 1754:1755 arg1 residue 1757:1763 residue 1757:1763 residue 1757:1763 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 7 53 from levels 1087:1092 arg1 cells 1119:1123 TM-treated cells 1108:1123 TM-treated cells 1108:1123 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 10 54 theme enzymatic 1675:1683 arg1 activity 1685:1692 enzymatic activity 1675:1692 enzymatic activity 1675:1692 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 8 55 theme Enzymatic 1200:1208 arg1 deglycosylation 1210:1224 Enzymatic deglycosylation 1200:1224 Enzymatic deglycosylation of purified recombinant SMPDL3A 1200:1256 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 10 56 theme Asn356 1514:1519 arg1 Glycosylation 1497:1509 Glycosylation 1497:1509 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, 1497:1593 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 5 57 theme CHO 874:876 arg1 cells 878:882 CHO cells 874:882 CHO cells expressing human SMPDL3A 874:907 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 8 58 theme phosphodiesterase 1295:1311 arg1 activity 1313:1320 phosphodiesterase activity 1295:1320 phosphodiesterase activity 1295:1320 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 6 59 gly non-glycosylated 963:978 arg1 SMPDL3A 980:986 non-glycosylated SMPDL3A 963:986 non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome 963:1052 Tunicamycin (TM) treatment resulted in expression of non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome. 28104755 8 60 theme recombinant 1238:1248 arg1 SMPDL3A 1250:1256 purified recombinant SMPDL3A 1229:1256 purified recombinant SMPDL3A 1229:1256 Enzymatic deglycosylation of purified recombinant SMPDL3A also resulted in significant loss of phosphodiesterase activity. 28104755 6 61 theme non-glycosylated 963:978 arg1 SMPDL3A 980:986 non-glycosylated SMPDL3A 963:986 non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome 963:1052 Tunicamycin (TM) treatment resulted in expression of non-glycosylated SMPDL3A that was not secreted, and was largely degraded by the proteasome. 28104755 9 62 gly glycosylation 1407:1419 arg1 Asn222 1434:1439 Asn222 1434:1439 Asn222 1434:1439 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 9 62 gly glycosylation 1407:1419 arg1 Asn69 1424:1428 Asn69 1424:1428 Asn69 1424:1428 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 7 63 theme Proteasomal 1055:1065 arg1 inhibition 1067:1076 Proteasomal inhibition 1055:1076 Proteasomal inhibition 1055:1076 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 4 64 theme structural 481:490 arg1 studies 492:498 recent structural studies 474:498 recent structural studies 474:498 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 12 65 from essential 1965:1973 arg1 conclusion 1920:1929 conclusion 1920:1929 conclusion 1920:1929 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 9 66 theme Site-directed 1323:1335 arg1 mutagenesis 1337:1347 Site-directed mutagenesis 1323:1347 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A 1323:1394 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 10 67 link N-linked 1536:1543 arg1 SMPDL3A 1524:1530 SMPDL3A 1524:1530 SMPDL3A 1524:1530 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 10 67 link N-linked 1536:1543 arg1 site 1545:1548 an N-linked site 1533:1548 an N-linked site conserved throughout the aSMase-like family 1533:1592 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 1 68 theme phosphodiesterase 146:162 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 68 theme phosphodiesterase 146:162 arg1 SMPDL3A 178:184 SMPDL3A 178:184 SMPDL3A 178:184 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 68 theme phosphodiesterase 146:162 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 68 theme phosphodiesterase 146:162 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 5 69 theme human 759:763 arg1 SMPDL3A 765:771 human SMPDL3A 759:771 human SMPDL3A 759:771 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 9 70 theme N-glycosylation 1363:1377 arg1 sites 1379:1383 individual N-glycosylation sites 1352:1383 individual N-glycosylation sites 1352:1383 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 4 71 theme substrate 536:544 arg1 preferences 546:556 these unexpected substrate preferences 519:556 these unexpected substrate preferences 519:556 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 2 72 dep homologous 292:301 arg1 cannot 356:361 cannot 356:361 cannot cleave sphingomyelin 356:382 SMPDL3A is highly homologous to acid sphingomyelinase (aSMase), but unlike aSMase cannot cleave sphingomyelin. 28104755 12 73 from conclusion 1920:1929 arg1 essential 1965:1973 essential 1965:1973 essential 1965:1973 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 5 74 theme N-glycosylation 794:808 arg1 inhibitors 780:789 inhibitors 780:789 inhibitors of N-glycosylation and site-directed mutagenesis 780:838 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 12 75 theme human 2034:2038 arg1 SMPDL3A 2040:2046 human SMPDL3A 2034:2046 human SMPDL3A 2034:2046 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 11 76 from peptide 1783:1789 arg1 SMPDL3A 1794:1800 SMPDL3A 1794:1800 SMPDL3A 1794:1800 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 0 77 theme sphingomyelin 25:37 arg1 phosphodiesterase 39:55 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 4 78 theme biology 589:595 arg1 aspects 570:576 many other aspects 559:576 many other aspects 559:576 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 0 79 theme acid-like 57:65 arg1 SMPDL3A 71:77 SMPDL3A 71:77 SMPDL3A 71:77 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 0 79 theme acid-like 57:65 arg1 3A 67:68 acid-like 3A 57:68 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 0 80 gly N-glycosylation 0:14 arg1 phosphodiesterase 39:55 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 19:78 N-glycosylation of human sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is essential for stability, secretion and activity. 28104755 10 81 theme aSMase-like 1575:1585 arg1 family 1587:1592 the aSMase-like family 1571:1592 the aSMase-like family 1571:1592 Glycosylation of Asn356 in SMPDL3A, an N-linked site conserved throughout the aSMase-like family, was critical for protection against proteasomal degradation and preservation of enzymatic activity. 28104755 7 82 theme non-glycosylated 1140:1155 arg1 protein 1157:1163 this non-glycosylated protein 1135:1163 this non-glycosylated protein 1135:1163 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 3 83 theme nucleotide 412:421 arg1 tri- 423:426 nucleotide tri- 412:426 nucleotide tri- 412:426 Rather, SMPDL3A hydrolyzes nucleotide tri- and diphosphates and their derivatives. 28104755 4 84 theme other 564:568 arg1 aspects 570:576 many other aspects 559:576 many other aspects 559:576 While recent structural studies have shed light on these unexpected substrate preferences, many other aspects of SMPDL3A biology, which may give insight into its function in vivo, remain obscure. 28104755 7 85 gly non-glycosylated 1140:1155 arg1 protein 1157:1163 this non-glycosylated protein 1135:1163 this non-glycosylated protein 1135:1163 Proteasomal inhibition restored levels of SMPDL3A in TM-treated cells, although this non-glycosylated protein lacked phosphodiesterase activity. 28104755 12 86 theme site-specific 1932:1944 arg1 N-glycosylation 1946:1960 site-specific N-glycosylation 1932:1960 site-specific N-glycosylation 1932:1960 In conclusion, site-specific N-glycosylation is essential for the intracellular stability, secretion and activity of human SMPDL3A. 28104755 5 87 from roles 689:693 arg1 secretion 733:741 secretion 733:741 secretion 733:741 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 5 87 from roles 689:693 arg1 expression 721:730 expression 721:730 expression 721:730 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 5 87 from roles 689:693 arg1 activity 747:754 activity 747:754 activity 747:754 Here, we investigate the roles of N-glycosylation in the expression, secretion and activity of human SMPDL3A, using inhibitors of N-glycosylation and site-directed mutagenesis, with either THP-1 macrophages or CHO cells expressing human SMPDL3A. 28104755 9 88 from mutagenesis 1337:1347 arg1 SMPDL3A 1388:1394 SMPDL3A 1388:1394 SMPDL3A 1388:1394 Site-directed mutagenesis of individual N-glycosylation sites in SMPDL3A identified glycosylation of Asn69 and Asn222 as affecting maturation of its N-glycans and secretion. 28104755 11 89 theme residue 1757:1763 arg1 peptide 1783:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 a predicted 22 residue N-terminal signal peptide 1742:1789 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 89 theme residue 1757:1763 arg1 essential 1812:1820 essential 1812:1820 essential 1812:1820 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 11 90 theme experimental 1716:1727 arg1 evidence 1729:1736 the first experimental evidence 1706:1736 the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells 1706:1914 We provide the first experimental evidence for a predicted 22 residue N-terminal signal peptide in SMPDL3A, which is essential for facilitating glycosylation and is removed from the mature protein secreted from CHO cells. 28104755 1 91 theme secreted 242:249 arg1 glycoprotein 260:271 a secreted N-linked glycoprotein 240:271 a secreted N-linked glycoprotein 240:271 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 91 theme secreted 242:249 arg1 phosphodiesterase 212:228 a recently identified phosphodiesterase 190:228 a recently identified phosphodiesterase 190:228 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28104755 1 91 theme secreted 242:249 arg1 3A 174:175 Sphingomyelin phosphodiesterase acid-like 3A 132:175 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) 132:185 Sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) is a recently identified phosphodiesterase, which is a secreted N-linked glycoprotein. 28322444 0 0 from characterization 19:34 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 1 1 theme family 152:157 arg1 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 1 theme family 152:157 arg1 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 5 2 theme channel 1067:1073 arg1 roles 1037:1041 trafficking-dependent roles 1015:1041 trafficking-dependent roles of the Kv3.1b potassium channel 1015:1073 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 3 gly N-glycosylation 284:298 arg1 residues 323:330 specific asparagine residues 303:330 specific asparagine residues 303:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 2 3 gly N-glycosylation 284:298 arg2 residues 323:330 specific asparagine residues 303:330 specific asparagine residues 303:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 5 4 gly N229-glycosylated 854:870 arg1 protein 879:885 the N229-glycosylated Kv3.1b protein 850:885 the N229-glycosylated Kv3.1b protein 850:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 5 from residues 323:330 arg1 N-glycosylation 284:298 N-glycosylation 284:298 N-glycosylation at specific asparagine residues 284:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 2 5 from residues 323:330 arg1 impact 274:279 the impact 270:279 the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein 270:371 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 4 6 theme located 694:700 arg1 pools 688:692 Kv3.1b pools 681:692 Kv3.1b pools located at the plasma membrane 681:723 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 0 7 from Identification 0:13 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 4 8 theme Kv3.1b 591:596 arg1 proteins 598:605 These ER-retained Kv3.1b proteins 573:605 These ER-retained Kv3.1b proteins 573:605 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 3 9 theme endoplasmic 545:555 arg1 ER 568:569 ER 568:569 ER 568:569 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 3 9 theme endoplasmic 545:555 arg1 reticulum 557:565 the endoplasmic reticulum 541:565 the endoplasmic reticulum (ER) 541:570 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 4 10 theme ER-retained 579:589 arg1 proteins 598:605 These ER-retained Kv3.1b proteins 573:605 These ER-retained Kv3.1b proteins 573:605 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 5 11 theme Mass 741:744 arg1 analysis 759:766 Mass spectrometry analysis 741:766 Mass spectrometry analysis 741:766 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 4 12 theme plasma 709:714 arg1 membrane 716:723 the plasma membrane 705:723 the plasma membrane 705:723 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 1 13 gly glycosylated 198:209 arg2 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 13 gly glycosylated 198:209 arg1 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 13 gly glycosylated 198:209 arg1 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 13 gly glycosylated 198:209 arg2 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 13 gly glycosylated 198:209 arg1 form 227:230 their mature form 214:230 their mature form 214:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 5 14 theme protein 879:885 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 14 theme protein 879:885 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 1 15 theme voltage-gated 162:174 arg1 channels 180:187 voltage-gated ion channels 162:187 voltage-gated ion channels 162:187 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 5 16 gly N220-glycosylated 946:962 arg1 Kv3.1b. 964:970 N220-glycosylated Kv3.1b. 946:970 N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 946:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 0 17 from N-glycosylation 53:67 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 18 theme type 787:790 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 18 theme type 787:790 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 19 theme high-mannose 904:915 arg1 glycan 935:940 a high-mannose type Man8 GlcNAc2 glycan 902:940 a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 902:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 3 20 gly unglycosylated 495:508 arg1 Kv3.1b 510:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 1 21 theme ion 176:178 arg1 channels 180:187 voltage-gated ion channels 162:187 voltage-gated ion channels 162:187 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 3 22 theme Large 374:378 arg1 quantities 380:389 Large quantities 374:389 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b 374:434 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 5 23 theme complex 779:785 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 23 theme complex 779:785 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 24 theme specific 303:310 arg1 residues 323:330 specific asparagine residues 303:330 specific asparagine residues 303:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 5 25 theme N-glycan 1129:1136 arg1 structure 1138:1146 N-glycan structure 1129:1146 N-glycan structure 1129:1146 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 1 26 theme channels 180:187 arg1 family 152:157 a family 150:157 a family of voltage-gated ion channels 150:187 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 5 27 theme N229-glycosylated 854:870 arg1 protein 879:885 the N229-glycosylated Kv3.1b protein 850:885 the N229-glycosylated Kv3.1b protein 850:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 1 28 theme potassium 106:114 arg1 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 28 theme potassium 106:114 arg1 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 2 29 theme protein 365:371 arg1 trafficking 339:349 the trafficking 335:349 the trafficking of the Kv3.1b protein 335:371 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 5 30 theme GlcNAc2 927:933 arg1 glycan 935:940 a high-mannose type Man8 GlcNAc2 glycan 902:940 a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 902:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 31 theme N-glycosylation 284:298 arg1 impact 274:279 the impact 270:279 the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein 270:371 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 3 32 theme plasma 448:453 arg1 membrane 455:462 the plasma membrane 444:462 the plasma membrane 444:462 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 3 33 gly N220-glycosylated 473:489 arg1 Kv3.1b 510:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 5 34 theme cell 1227:1230 arg1 expression 1240:1249 cell surface expression 1227:1249 cell surface expression 1227:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 35 theme N220-glycosylated 946:962 arg1 Kv3.1b. 964:970 N220-glycosylated Kv3.1b. 946:970 N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 946:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 1 36 theme channel 120:126 arg1 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 36 theme channel 120:126 arg1 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 5 37 theme Kv3.1b 872:877 arg1 protein 879:885 the N229-glycosylated Kv3.1b protein 850:885 the N229-glycosylated Kv3.1b protein 850:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 0 38 theme N-glycosylation 53:67 arg1 characterization 19:34 characterization 19:34 characterization 19:34 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 0 38 theme N-glycosylation 53:67 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 39 theme surface 1232:1238 arg1 expression 1240:1249 cell surface expression 1227:1249 cell surface expression 1227:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 0 40 from Kv3.1b 94:99 arg1 characterization 19:34 characterization 19:34 characterization 19:34 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 0 40 from Kv3.1b 94:99 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 41 theme trafficking-dependent 1015:1035 arg1 roles 1037:1041 trafficking-dependent roles 1015:1041 trafficking-dependent roles of the Kv3.1b potassium channel 1015:1073 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 0 42 theme site-specific 39:51 arg1 N-glycosylation 53:67 site-specific N-glycosylation 39:67 site-specific N-glycosylation in the potassium channel Kv3.1b 39:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 43 theme Hex3 792:795 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 43 theme Hex3 792:795 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 44 theme site-specific 1097:1109 arg1 glycosylation 1111:1123 N229 site-specific glycosylation 1092:1123 N229 site-specific glycosylation 1092:1123 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 45 theme spectrometry 746:757 arg1 analysis 759:766 Mass spectrometry analysis 741:766 Mass spectrometry analysis 741:766 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 46 theme HexNAc4 797:803 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 46 theme HexNAc4 797:803 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 3 47 theme unglycosylated 495:508 arg1 Kv3.1b 510:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 5 48 theme Fuc1 805:808 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 48 theme Fuc1 805:808 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 3 49 theme -glycosylated 415:427 arg1 Kv3.1b 429:434 asparagine 229 (N229)-glycosylated Kv3.1b 394:434 asparagine 229 (N229)-glycosylated Kv3.1b 394:434 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 3 50 theme N220-glycosylated 473:489 arg1 Kv3.1b 510:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 N220-glycosylated and unglycosylated Kv3.1b 473:515 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 3 51 gly -glycosylated 415:427 arg1 Kv3.1b 429:434 asparagine 229 (N229)-glycosylated Kv3.1b 394:434 asparagine 229 (N229)-glycosylated Kv3.1b 394:434 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 5 52 theme specific 1189:1196 arg1 structures 1207:1216 specific N-glycan structures 1189:1216 specific N-glycan structures 1189:1216 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 53 theme potassium 1057:1065 arg1 channel 1067:1073 the Kv3.1b potassium channel 1046:1073 the Kv3.1b potassium channel 1046:1073 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 54 theme Kv3.1b 1050:1055 arg1 channel 1067:1073 the Kv3.1b potassium channel 1046:1073 the Kv3.1b potassium channel 1046:1073 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 55 theme type 917:920 arg1 glycan 935:940 a high-mannose type Man8 GlcNAc2 glycan 902:940 a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 902:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 56 theme N-glycan 1198:1205 arg1 structures 1207:1216 specific N-glycan structures 1189:1216 specific N-glycan structures 1189:1216 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 4 57 dep susceptible 612:622 arg1 whereas 673:679 whereas 673:679 whereas 673:679 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 3 58 dep reached 436:442 arg1 whereas 465:471 whereas 465:471 whereas 465:471 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 5 59 theme Man8 922:925 arg1 glycan 935:940 a high-mannose type Man8 GlcNAc2 glycan 902:940 a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression 902:1249 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 1 60 theme ion 116:118 arg1 Kv3.1b 128:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b 102:133 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 60 theme ion 116:118 arg1 member 140:145 a member 138:145 a member of a family of voltage-gated ion channels that are glycosylated in their mature form 138:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 1 61 theme mature 220:225 arg1 form 227:230 their mature form 214:230 their mature form 214:230 The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. 28322444 4 62 from membrane 716:723 arg1 located 694:700 located 694:700 located 694:700 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 28322444 0 63 gly N-glycosylation 53:67 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 64 theme N229 1092:1095 arg1 glycosylation 1111:1123 N229 site-specific glycosylation 1092:1123 N229 site-specific glycosylation 1092:1123 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 65 theme asparagine 312:321 arg1 residues 323:330 specific asparagine residues 303:330 specific asparagine residues 303:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 2 66 from impact 274:279 arg1 residues 323:330 specific asparagine residues 303:330 specific asparagine residues 303:330 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 2 66 from impact 274:279 arg1 trafficking 339:349 the trafficking 335:349 the trafficking of the Kv3.1b protein 335:371 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 0 67 theme channel 86:92 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 2 68 theme present 240:246 arg1 study 248:252 the present study 236:252 the present study 236:252 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 5 69 theme glycan 830:835 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 69 theme glycan 830:835 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 2 70 theme Kv3.1b 358:363 arg1 protein 365:371 the Kv3.1b protein 354:371 the Kv3.1b protein 354:371 In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. 28322444 3 71 theme Kv3.1b 429:434 arg1 quantities 380:389 Large quantities 374:389 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b 374:434 Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). 28322444 0 72 theme potassium 76:84 arg1 Kv3.1b 94:99 the potassium channel Kv3.1b 72:99 the potassium channel Kv3.1b 72:99 Identification and characterization of site-specific N-glycosylation in the potassium channel Kv3.1b. 28322444 5 73 theme major 824:828 arg1 component 837:845 the major glycan component 820:845 the major glycan component of the N229-glycosylated Kv3.1b protein 820:885 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 5 73 theme major 824:828 arg1 glycan 810:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 a complex type Hex3 HexNAc4 Fuc1 glycan 777:815 Mass spectrometry analysis revealed a complex type Hex3 HexNAc4 Fuc1 glycan as the major glycan component of the N229-glycosylated Kv3.1b protein, as opposed to a high-mannose type Man8 GlcNAc2 glycan for N220-glycosylated Kv3.1b. Taken together, these results suggest that trafficking-dependent roles of the Kv3.1b potassium channel are dependent on N229 site-specific glycosylation and N-glycan structure, and operate through a mechanism whereby specific N-glycan structures regulate cell surface expression. 28322444 4 74 theme Kv3.1b 681:686 arg1 pools 688:692 Kv3.1b pools 681:692 Kv3.1b pools located at the plasma membrane 681:723 These ER-retained Kv3.1b proteins were susceptible to degradation, when co-expressed with calnexin, whereas Kv3.1b pools located at the plasma membrane were resistant. 29233911 11 0 theme glycosylation 1450:1462 arg1 characterization 1423:1438 The comprehensive characterization 1405:1438 The comprehensive characterization of C1-Inh glycosylation described in this study 1405:1486 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 4 1 theme electron-transfer 723:739 arg1 ETD 755:757 ETD 755:757 ETD 755:757 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 1 theme electron-transfer 723:739 arg1 dissociation 741:752 electron-transfer dissociation 723:752 electron-transfer dissociation (ETD) 723:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 6 2 gly O-glycosylated 937:950 arg1 region 963:968 an extensively O-glycosylated N-terminal region 922:968 an extensively O-glycosylated N-terminal region 922:968 The analysis revealed an extensively O-glycosylated N-terminal region. 29233911 11 3 theme functional 1520:1529 arg1 studies 1531:1537 further functional studies 1512:1537 further functional studies on the role of these glycan modifications 1512:1579 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 11 4 theme comprehensive 1409:1421 arg1 characterization 1423:1438 The comprehensive characterization 1405:1438 The comprehensive characterization of C1-Inh glycosylation described in this study 1405:1486 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 9 5 theme known 1207:1211 arg1 sites 1233:1237 all known six N-glycosylation sites 1203:1237 all known six N-glycosylation sites 1203:1237 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 1 6 theme lectin 246:251 arg1 pathways 264:271 the classical and lectin complement pathways 228:271 pathways 264:271 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 4 7 theme site-specific 488:500 arg1 N- 502:503 an in-depth site-specific N- and O-glycosylation analysis 476:532 N- 502:503 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 7 8 theme known 991:995 arg1 sites 1013:1017 five known O-glycosylation sites 986:1017 five known O-glycosylation sites 986:1017 Five novel and five known O-glycosylation sites were identified, carrying mainly core1-type O-glycans. 29233911 4 9 theme collision-induced 682:698 arg1 CID 714:716 CID 714:716 CID 714:716 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 9 theme collision-induced 682:698 arg1 dissociation 700:711 stepping-energy collision-induced dissociation 666:711 stepping-energy collision-induced dissociation (CID) 666:717 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 1 10 theme complement 253:262 arg1 pathways 264:271 the classical and lectin complement pathways 228:271 pathways 264:271 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 5 11 theme Various 761:767 arg1 proteases 769:777 Various proteases 761:777 Various proteases 761:777 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 9 12 theme N-glycosylation 1217:1231 arg1 sites 1233:1237 all known six N-glycosylation sites 1203:1237 all known six N-glycosylation sites 1203:1237 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 11 13 theme further 1512:1518 arg1 studies 1531:1537 further functional studies 1512:1537 further functional studies on the role of these glycan modifications 1512:1579 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 1 14 theme serine 133:138 arg1 inhibitor 149:157 a serine protease inhibitor 131:157 a serine protease inhibitor 131:157 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 14 theme serine 133:138 arg1 C1-inhibitor 106:117 Human C1-inhibitor 100:117 Human C1-inhibitor (C1-Inh) 100:126 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 4 15 theme PGC 634:636 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 5 16 with combination 803:813 arg1 F 827:827 PNGase F 820:827 PNGase F 820:827 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 5 16 with combination 803:813 arg1 treatment 848:856 exoglycosidase treatment 833:856 exoglycosidase treatment 833:856 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 8 17 gly O-glycosylated 1109:1122 arg1 portion 1124:1130 a heavily O-glycosylated portion 1099:1130 a heavily O-glycosylated portion spanning from Thr82-Ser121 1099:1157 In addition, we detected a heavily O-glycosylated portion spanning from Thr82-Ser121 with up to 16 O-glycans attached. 29233911 1 18 theme major 167:171 arg1 regulator 173:181 the major regulator 163:181 the major regulator of the contact activation pathway 163:215 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 3 19 theme C1-Inh 396:401 arg1 glycosylation 403:415 C1-Inh glycosylation 396:415 C1-Inh glycosylation 396:415 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 3 20 theme biological 377:386 arg1 role 388:391 biological role 377:391 biological role 377:391 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 4 21 theme stepping-energy 666:680 arg1 CID 714:716 CID 714:716 CID 714:716 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 21 theme stepping-energy 666:680 arg1 dissociation 700:711 stepping-energy collision-induced dissociation 666:711 stepping-energy collision-induced dissociation (CID) 666:717 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 22 theme first 465:469 arg1 time 471:474 the first time 461:474 the first time 461:474 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 8 23 dep 16 1170:1171 arg1 to 1167:1168 to 1167:1168 to 1167:1168 In addition, we detected a heavily O-glycosylated portion spanning from Thr82-Ser121 with up to 16 O-glycans attached. 29233911 1 24 theme Human 100:104 arg1 C1-inhibitor 106:117 Human C1-inhibitor 100:117 Human C1-inhibitor (C1-Inh) 100:126 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 24 theme Human 100:104 arg1 inhibitor 149:157 a serine protease inhibitor 131:157 a serine protease inhibitor 131:157 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 24 theme Human 100:104 arg1 C1-Inh 120:125 C1-Inh 120:125 C1-Inh 120:125 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 5 25 theme glyco 884:888 arg1 peptides 890:897 the (glyco)peptides 879:897 the (glyco)peptides 879:897 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 0 26 theme N- 0:1 arg1 Analysis 23:30 N- and O-glycosylation Analysis 0:30 N- and O-glycosylation Analysis of Human C1-inhibitor 0:52 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 5 27 theme exoglycosidase 833:846 arg1 treatment 848:856 exoglycosidase treatment 833:856 exoglycosidase treatment 833:856 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 3 28 theme structural 353:362 arg1 unknown 429:435 unknown 429:435 unknown 429:435 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 3 28 theme structural 353:362 arg1 features 364:371 the structural features 349:371 the structural features 349:371 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 4 29 theme C1-Inh 537:542 arg1 analysis 525:532 an in-depth site-specific N- and O-glycosylation analysis 476:532 analysis 525:532 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 29 theme C1-Inh 537:542 arg1 N- 502:503 an in-depth site-specific N- and O-glycosylation analysis 476:532 N- 502:503 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 8 30 theme O-glycosylated 1109:1122 arg1 portion 1124:1130 a heavily O-glycosylated portion 1099:1130 a heavily O-glycosylated portion spanning from Thr82-Ser121 1099:1157 In addition, we detected a heavily O-glycosylated portion spanning from Thr82-Ser121 with up to 16 O-glycans attached. 29233911 10 31 theme released 1362:1369 arg1 N-glycans 1371:1379 released N-glycans 1362:1379 released N-glycans by MALDI-TOF/TOF-MS/MS 1362:1402 The glycoforms were in accordance with results on released N-glycans by MALDI-TOF/TOF-MS/MS. 29233911 1 32 theme contact 190:196 arg1 pathway 209:215 the contact activation pathway 186:215 the contact activation pathway 186:215 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 0 33 theme O-glycosylation 7:21 arg1 Analysis 23:30 N- and O-glycosylation Analysis 0:30 N- and O-glycosylation Analysis of Human C1-inhibitor 0:52 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 4 34 theme mass 562:565 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 34 theme mass 562:565 arg1 approaches 581:590 various mass spectrometric approaches 554:590 various mass spectrometric approaches 554:590 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 1 35 theme activation 198:207 arg1 pathway 209:215 the contact activation pathway 186:215 the contact activation pathway 186:215 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 0 36 theme Human 35:39 arg1 C1-inhibitor 41:52 Human C1-inhibitor 35:52 Human C1-inhibitor 35:52 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 4 37 theme various 554:560 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 37 theme various 554:560 arg1 approaches 581:590 various mass spectrometric approaches 554:590 various mass spectrometric approaches 554:590 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 11 38 from studies 1531:1537 arg1 role 1546:1549 the role 1542:1549 the role of these glycan modifications 1542:1579 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 1 39 theme pathway 209:215 arg1 inhibitor 149:157 a serine protease inhibitor 131:157 a serine protease inhibitor 131:157 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 39 theme pathway 209:215 arg1 regulator 173:181 the major regulator 163:181 the major regulator of the contact activation pathway 163:215 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 39 theme pathway 209:215 arg1 C1-inhibitor 106:117 Human C1-inhibitor 100:117 Human C1-inhibitor (C1-Inh) 100:126 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 39 theme pathway 209:215 arg1 pathways 264:271 the classical and lectin complement pathways 228:271 pathways 264:271 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 3 40 theme glycosylation 403:415 arg1 unknown 429:435 unknown 429:435 unknown 429:435 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 3 40 theme glycosylation 403:415 arg1 features 364:371 the structural features 349:371 the structural features 349:371 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 3 40 theme glycosylation 403:415 arg1 role 388:391 biological role 377:391 biological role 377:391 However, both the structural features and biological role of C1-Inh glycosylation are largely unknown. 29233911 4 41 theme O-glycosylation 509:523 arg1 analysis 525:532 an in-depth site-specific N- and O-glycosylation analysis 476:532 analysis 525:532 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 42 theme spectrometric 567:579 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 4 42 theme spectrometric 567:579 arg1 approaches 581:590 various mass spectrometric approaches 554:590 various mass spectrometric approaches 554:590 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 9 43 theme site-specific 1274:1286 arg1 analysis 1302:1309 this site-specific glycosylation analysis 1269:1309 this site-specific glycosylation analysis 1269:1309 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 5 44 theme PNGase 820:825 arg1 F 827:827 PNGase F 820:827 PNGase F 820:827 Various proteases were applied, partly in combination with PNGase F and exoglycosidase treatment, in order to analyze the (glyco)peptides. 29233911 2 45 gly glycoprotein 321:332 arg1 glycoprotein 321:332 a highly glycosylated plasma glycoprotein 292:332 a highly glycosylated plasma glycoprotein 292:332 It is known to be a highly glycosylated plasma glycoprotein. 29233911 2 45 gly glycoprotein 321:332 arg1 It 274:275 It 274:275 It 274:275 It is known to be a highly glycosylated plasma glycoprotein. 29233911 9 46 theme glycosylation 1288:1300 arg1 analysis 1302:1309 this site-specific glycosylation analysis 1269:1309 this site-specific glycosylation analysis 1269:1309 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 0 47 gly O-glycosylation 7:21 arg1 C1-inhibitor 41:52 Human C1-inhibitor 35:52 Human C1-inhibitor 35:52 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 7 48 theme core1-type 1052:1061 arg1 O-glycans 1063:1071 mainly core1-type O-glycans 1045:1071 mainly core1-type O-glycans 1045:1071 Five novel and five known O-glycosylation sites were identified, carrying mainly core1-type O-glycans. 29233911 0 49 theme C1-inhibitor 41:52 arg1 Analysis 23:30 N- and O-glycosylation Analysis 0:30 N- and O-glycosylation Analysis of Human C1-inhibitor 0:52 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 6 50 theme O-glycosylated 937:950 arg1 region 963:968 an extensively O-glycosylated N-terminal region 922:968 an extensively O-glycosylated N-terminal region 922:968 The analysis revealed an extensively O-glycosylated N-terminal region. 29233911 11 51 theme glycan 1560:1565 arg1 modifications 1567:1579 these glycan modifications 1554:1579 these glycan modifications 1554:1579 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 6 52 theme N-terminal 952:961 arg1 region 963:968 an extensively O-glycosylated N-terminal region 922:968 an extensively O-glycosylated N-terminal region 922:968 The analysis revealed an extensively O-glycosylated N-terminal region. 29233911 2 53 gly glycosylated 301:312 arg1 glycoprotein 321:332 a highly glycosylated plasma glycoprotein 292:332 a highly glycosylated plasma glycoprotein 292:332 It is known to be a highly glycosylated plasma glycoprotein. 29233911 2 53 gly glycosylated 301:312 arg1 It 274:275 It 274:275 It 274:275 It is known to be a highly glycosylated plasma glycoprotein. 29233911 2 54 theme plasma 314:319 arg1 glycoprotein 321:332 a highly glycosylated plasma glycoprotein 292:332 a highly glycosylated plasma glycoprotein 292:332 It is known to be a highly glycosylated plasma glycoprotein. 29233911 2 54 theme plasma 314:319 arg1 It 274:275 It 274:275 It 274:275 It is known to be a highly glycosylated plasma glycoprotein. 29233911 11 55 theme modifications 1567:1579 arg1 role 1546:1549 the role 1542:1549 the role of these glycan modifications 1542:1579 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 4 56 theme graphitized 614:624 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 10 57 from results 1351:1357 arg1 N-glycans 1371:1379 released N-glycans 1362:1379 released N-glycans by MALDI-TOF/TOF-MS/MS 1362:1402 The glycoforms were in accordance with results on released N-glycans by MALDI-TOF/TOF-MS/MS. 29233911 11 58 theme C1-Inh 1443:1448 arg1 glycosylation 1450:1462 C1-Inh glycosylation 1443:1462 C1-Inh glycosylation 1443:1462 The comprehensive characterization of C1-Inh glycosylation described in this study will form the basis for further functional studies on the role of these glycan modifications. 29233911 2 59 theme glycosylated 301:312 arg1 glycoprotein 321:332 a highly glycosylated plasma glycoprotein 292:332 a highly glycosylated plasma glycoprotein 292:332 It is known to be a highly glycosylated plasma glycoprotein. 29233911 2 59 theme glycosylated 301:312 arg1 It 274:275 It 274:275 It 274:275 It is known to be a highly glycosylated plasma glycoprotein. 29233911 4 60 theme C18-porous 603:612 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 0 61 theme Mucin-type 72:81 arg1 O-Glycosylation 83:97 Extensive Mucin-type O-Glycosylation 62:97 Extensive Mucin-type O-Glycosylation 62:97 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 4 62 gly O-glycosylation 509:523 arg1 C1-Inh 537:542 C1-Inh 537:542 C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 537:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 1 63 theme protease 140:147 arg1 inhibitor 149:157 a serine protease inhibitor 131:157 a serine protease inhibitor 131:157 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 1 63 theme protease 140:147 arg1 C1-inhibitor 106:117 Human C1-inhibitor 100:117 Human C1-inhibitor (C1-Inh) 100:126 Human C1-inhibitor (C1-Inh) is a serine protease inhibitor and the major regulator of the contact activation pathway as well as the classical and lectin complement pathways. 29233911 0 64 theme Extensive 62:70 arg1 O-Glycosylation 83:97 Extensive Mucin-type O-Glycosylation 62:97 Extensive Mucin-type O-Glycosylation 62:97 N- and O-glycosylation Analysis of Human C1-inhibitor Reveals Extensive Mucin-type O-Glycosylation. 29233911 4 65 theme in-depth 479:486 arg1 N- 502:503 an in-depth site-specific N- and O-glycosylation analysis 476:532 N- 502:503 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 29233911 9 66 gly N-glycosylation 1217:1231 arg2 sites 1233:1237 all known six N-glycosylation sites 1203:1237 all known six N-glycosylation sites 1203:1237 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 9 66 gly N-glycosylation 1217:1231 arg2 six 1213:1215 six 1213:1215 six 1213:1215 Likewise, all known six N-glycosylation sites were covered and confirmed by this site-specific glycosylation analysis. 29233911 7 67 theme O-glycosylation 997:1011 arg1 sites 1013:1017 five known O-glycosylation sites 986:1017 five known O-glycosylation sites 986:1017 Five novel and five known O-glycosylation sites were identified, carrying mainly core1-type O-glycans. 29233911 7 68 gly O-glycosylation 997:1011 arg2 five 986:989 five 986:989 five 986:989 Five novel and five known O-glycosylation sites were identified, carrying mainly core1-type O-glycans. 29233911 7 68 gly O-glycosylation 997:1011 arg2 sites 1013:1017 five known O-glycosylation sites 986:1017 five known O-glycosylation sites 986:1017 Five novel and five known O-glycosylation sites were identified, carrying mainly core1-type O-glycans. 29233911 4 69 theme carbon 626:631 arg1 -LC-ESI-QTOF-MS/MS 638:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS 603:655 C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD) 603:758 Here, we performed for the first time an in-depth site-specific N- and O-glycosylation analysis of C1-Inh combining various mass spectrometric approaches, including C18-porous graphitized carbon (PGC)-LC-ESI-QTOF-MS/MS applying stepping-energy collision-induced dissociation (CID) and electron-transfer dissociation (ETD). 28630087 12 0 theme complex 2136:2142 arg1 formation 2144:2152 concentration-dependent complex formation 2112:2152 concentration-dependent complex formation 2112:2152 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 6 1 theme site 1275:1278 arg1 accessibility 1288:1300 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 3 2 theme receptor 765:772 arg1 interaction 774:784 receptor interaction 765:784 receptor interaction 765:784 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 10 3 theme glycosylation 1662:1674 arg1 located 1694:1700 located 1694:1700 located 1694:1700 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 3 theme glycosylation 1662:1674 arg1 HNE 1685:1687 HNE 1685:1687 HNE 1685:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 3 theme glycosylation 1662:1674 arg1 sites 1676:1680 The three glycosylation sites 1652:1680 The three glycosylation sites of HNE 1652:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 3 4 theme spatiotemporal 572:585 arg1 expression 587:596 The spatiotemporal expression 568:596 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation 568:685 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 14 5 theme multiple 2610:2617 arg1 functions 2626:2634 multiple immune functions 2610:2634 multiple immune functions central to inflammation and infection 2610:2672 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 13 6 theme aeruginosa 2382:2391 arg1 inhibition 2336:2345 low micromolar concentration-dependent growth inhibition 2290:2345 low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa 2290:2391 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 1 7 theme N-glycosylation 287:301 arg1 structure 238:246 structure 238:246 structure 238:246 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 7 theme N-glycosylation 287:301 arg1 functions 270:278 immune-modulating functions 252:278 immune-modulating functions 252:278 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 9 8 from regulation 1545:1554 arg1 neutrophils 1639:1649 activated human neutrophils 1623:1649 activated human neutrophils 1623:1649 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 13 9 theme concentration-dependent 2305:2327 arg1 inhibition 2336:2345 low micromolar concentration-dependent growth inhibition 2290:2345 low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa 2290:2391 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 11 10 theme paucimannosidic 1824:1838 arg1 glycoforms 1844:1853 the paucimannosidic HNE glycoforms 1820:1853 the paucimannosidic HNE glycoforms 1820:1853 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 2 11 theme glycan 346:351 arg1 profiling 384:392 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 profiling 384:392 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 1 12 theme innate 208:213 arg1 system 222:227 the innate immune system 204:227 the innate immune system 204:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 8 13 theme HNE 1419:1421 arg1 residing 1423:1430 HNE residing 1419:1430 HNE residing 1419:1430 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 5 14 gly fucosylated 1044:1054 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 12 15 theme HNE 2062:2064 arg1 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 12 15 theme HNE 2062:2064 arg1 inhibitor 2075:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 11 16 gly sialoglycoforms 1940:1954 arg1 HNE 1936:1938 the HNE sialoglycoforms 1932:1954 the HNE sialoglycoforms 1932:1954 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 3 17 theme growth 821:826 arg1 assays 828:833 growth assays 821:833 growth assays 821:833 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 12 18 gly N-glycosylated 2047:2060 arg1 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 12 18 gly N-glycosylated 2047:2060 arg1 inhibitor 2075:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 9 19 theme Stimulation-induced 1469:1487 arg1 mobilization 1502:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 3 20 dep immunoblotting 716:729 arg1 assays 828:833 growth assays 821:833 growth assays 821:833 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 8 21 theme neutrophil 1444:1453 arg1 compartments 1455:1466 distinct neutrophil compartments 1435:1466 distinct neutrophil compartments 1435:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 1 22 dep structure 238:246 arg1 the 234:236 the 234:236 the 234:236 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 9 23 theme surface 1494:1500 arg1 mobilization 1502:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 5 24 theme fucosylated 1044:1054 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 1 25 gly N-glycosylated 170:183 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 25 gly N-glycosylated 170:183 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 5 26 theme complex-type 1068:1079 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 10 27 with interference 1767:1778 arg1 activity 1796:1803 HNE enzyme activity 1785:1803 HNE enzyme activity 1785:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 6 28 theme unoccupied 1167:1176 arg1 Asn88 1178:1182 the mostly unoccupied Asn88 1156:1182 the mostly unoccupied Asn88 1156:1182 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 13 29 theme micromolar 2294:2303 arg1 inhibition 2336:2345 low micromolar concentration-dependent growth inhibition 2290:2345 low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa 2290:2391 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 11 30 gly glycoforms 1844:1853 arg1 HNE 1840:1842 the paucimannosidic HNE glycoforms 1820:1853 the paucimannosidic HNE glycoforms 1820:1853 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 9 31 theme spatiotemporal 1530:1543 arg1 regulation 1545:1554 a spatiotemporal regulation 1528:1554 a spatiotemporal regulation 1528:1554 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 1 32 from protease 192:199 arg1 system 222:227 the innate immune system 204:227 the innate immune system 204:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 33 theme important 160:168 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 33 theme important 160:168 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 2 34 theme healthy 547:553 arg1 individuals 555:565 healthy individuals 547:565 healthy individuals 547:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 0 35 theme Immune 104:109 arg1 Functions 111:119 Its Immune Functions 100:119 Its Immune Functions 100:119 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 1 36 theme serine 185:190 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 36 theme serine 185:190 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 11 37 theme HNE 2005:2007 arg1 involvement 1990:2000 a glycoform-dependent involvement 1968:2000 a glycoform-dependent involvement of HNE in complement activation 1968:2032 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 11 38 from involvement 1990:2000 arg1 activation 2023:2032 complement activation 2012:2032 complement activation 2012:2032 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 6 39 gly glycosylation 1261:1273 arg2 accessibility 1288:1300 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 6 39 gly glycosylation 1261:1273 arg2 site 1275:1278 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 2 40 used utilized 399:406 arg2 profiling 384:392 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 profiling 384:392 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 13 41 theme HNE 2263:2265 arg1 N-glycans 2267:2275 isolated HNE N-glycans 2254:2275 isolated HNE N-glycans 2254:2275 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 13 42 theme HNE 2454:2456 arg1 N-glycans 2458:2466 the HNE N-glycans 2450:2466 the HNE N-glycans 2450:2466 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 10 43 theme glycan 1739:1744 arg1 functions 1746:1754 glycan functions 1739:1754 glycan functions other than interference with HNE enzyme activity 1739:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 9 44 theme surface-specific 1570:1585 arg1 signatures 1601:1610 cell surface-specific glycosylation signatures 1565:1610 cell surface-specific glycosylation signatures 1565:1610 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 13 45 theme active 2239:2244 arg1 HNE 2246:2248 enzymatically active HNE 2225:2248 enzymatically active HNE 2225:2248 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 0 46 theme Paucimannose-Rich 0:16 arg1 N-glycosylation 18:32 Paucimannose-Rich N-glycosylation 0:32 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase 0:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 1 47 theme Human 122:126 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 47 theme Human 122:126 arg1 HNE 149:151 HNE 149:151 HNE 149:151 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 47 theme Human 122:126 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 4 48 theme Site-specific 836:848 arg1 glycoprofiling 854:867 Site-specific HNE glycoprofiling 836:867 Site-specific HNE glycoprofiling 836:867 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 4 49 used occupied 991:998 arg2 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 4 49 used occupied 991:998 arg2 N-glycans 911:919 unusual paucimannosidic N-glycans 887:919 unusual paucimannosidic N-glycans 887:919 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 6 50 theme paucimannosidic 1207:1221 arg1 N-glycans 1223:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 10 51 theme other 1756:1760 arg1 functions 1746:1754 glycan functions 1739:1754 glycan functions other than interference with HNE enzyme activity 1739:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 2 52 theme HNE 464:466 arg1 N-glycosylation 445:459 the heterogeneous N-glycosylation 427:459 the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals 427:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 0 53 theme Regulated 54:62 arg1 Elastase 81:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 10 54 gly glycosylation 1662:1674 arg2 located 1694:1700 located 1694:1700 located 1694:1700 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 54 gly glycosylation 1662:1674 arg2 HNE 1685:1687 HNE 1685:1687 HNE 1685:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 54 gly glycosylation 1662:1674 arg1 HNE 1685:1687 HNE 1685:1687 HNE 1685:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 54 gly glycosylation 1662:1674 arg2 sites 1676:1680 The three glycosylation sites 1652:1680 The three glycosylation sites of HNE 1652:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 54 gly glycosylation 1662:1674 arg2 three 1656:1660 three 1656:1660 three 1656:1660 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 0 55 gly N-glycosylation 18:32 arg1 Elastase 81:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 12 56 theme preferred 2158:2166 arg1 interactions 2188:2199 preferred glycoform-glycoform interactions 2158:2199 preferred glycoform-glycoform interactions 2158:2199 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 2 57 theme isolated 515:522 arg1 granules 535:542 isolated neutrophil granules 515:542 isolated neutrophil granules of healthy individuals 515:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 2 58 gly glycoprotein 371:382 arg1 glycoprotein 371:382 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 glycoprotein 371:382 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 3 59 theme cell 732:735 arg1 capture 745:751 cell surface capture 732:751 cell surface capture 732:751 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 11 60 theme human 1889:1893 arg1 lectin 1911:1916 human mannose binding lectin 1889:1916 human mannose binding lectin 1889:1916 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 0 61 theme Neutrophil 70:79 arg1 Elastase 81:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 9 62 theme human 1633:1637 arg1 neutrophils 1639:1649 activated human neutrophils 1623:1649 activated human neutrophils 1623:1649 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 2 63 gly glycopeptide 354:365 arg2 glycopeptide 354:365 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 glycopeptide 354:365 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 4 64 theme paucimannosidic 895:909 arg1 N-glycans 911:919 unusual paucimannosidic N-glycans 887:919 unusual paucimannosidic N-glycans 887:919 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 4 64 theme paucimannosidic 895:909 arg1 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 12 65 theme concentration-dependent 2112:2134 arg1 formation 2144:2152 concentration-dependent complex formation 2112:2152 concentration-dependent complex formation 2112:2152 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 3 66 theme native 754:759 arg1 MS 761:762 native MS 754:762 native MS 754:762 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 6 67 theme solvent 1280:1286 arg1 accessibility 1288:1300 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 14 68 theme central 2636:2642 arg1 functions 2626:2634 multiple immune functions 2610:2634 multiple immune functions central to inflammation and infection 2610:2672 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 3 69 theme protease 787:794 arg1 inhibition 796:805 protease inhibition 787:805 protease inhibition 787:805 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 11 70 theme binding 1903:1909 arg1 lectin 1911:1916 human mannose binding lectin 1889:1916 human mannose binding lectin 1889:1916 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 8 71 theme Subcellular- 1332:1343 arg1 glycoprofiling 1363:1376 Subcellular- and site-specific glycoprofiling 1332:1376 Subcellular- and site-specific glycoprofiling 1332:1376 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 8 72 from N-glycosylation 1400:1414 arg1 compartments 1455:1466 distinct neutrophil compartments 1435:1466 distinct neutrophil compartments 1435:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 6 73 theme glycosylation 1261:1273 arg1 accessibility 1288:1300 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 14 74 theme immune 2619:2624 arg1 functions 2626:2634 multiple immune functions 2610:2634 multiple immune functions central to inflammation and infection 2610:2672 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 13 75 theme growth 2329:2334 arg1 inhibition 2336:2345 low micromolar concentration-dependent growth inhibition 2290:2345 low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa 2290:2391 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 8 76 gly N-glycosylation 1400:1414 arg1 compartments 1455:1466 distinct neutrophil compartments 1435:1466 distinct neutrophil compartments 1435:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 8 76 gly N-glycosylation 1400:1414 arg1 residing 1423:1430 HNE residing 1419:1430 HNE residing 1419:1430 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 3 77 theme neutrophil 612:621 arg1 activation 623:632 neutrophil activation 612:632 neutrophil activation 612:632 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 1 78 theme HNE 283:285 arg1 N-glycosylation 287:301 HNE N-glycosylation 283:301 HNE N-glycosylation 283:301 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 8 79 theme site-specific 1349:1361 arg1 glycoprofiling 1363:1376 Subcellular- and site-specific glycoprofiling 1332:1376 Subcellular- and site-specific glycoprofiling 1332:1376 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 11 80 theme glycoform-dependent 1970:1988 arg1 involvement 1990:2000 a glycoform-dependent involvement 1968:2000 a glycoform-dependent involvement of HNE in complement activation 1968:2032 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 3 81 theme HNE 601:603 arg1 expression 587:596 The spatiotemporal expression 568:596 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation 568:685 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 12 82 theme N-glycosylated 2047:2060 arg1 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 12 82 theme N-glycosylated 2047:2060 arg1 inhibitor 2075:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 10 83 theme HNE 1685:1687 arg1 located 1694:1700 located 1694:1700 located 1694:1700 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 83 theme HNE 1685:1687 arg1 HNE 1685:1687 HNE 1685:1687 HNE 1685:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 10 83 theme HNE 1685:1687 arg1 sites 1676:1680 The three glycosylation sites 1652:1680 The three glycosylation sites of HNE 1652:1687 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 3 84 theme biological 642:651 arg1 importance 653:662 the biological importance 638:662 the biological importance of its N-glycosylation 638:685 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 8 85 theme uniform 1392:1398 arg1 N-glycosylation 1400:1414 highly uniform N-glycosylation 1385:1414 highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments 1385:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 12 86 theme protease 2066:2073 arg1 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 α1-antitrypsin 2086:2099 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 12 86 theme protease 2066:2073 arg1 inhibitor 2075:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor 2035:2083 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 11 87 theme HNE 1840:1842 arg1 glycoforms 1844:1853 the paucimannosidic HNE glycoforms 1820:1853 the paucimannosidic HNE glycoforms 1820:1853 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 2 88 theme LC-MS/MS-based 331:344 arg1 profiling 384:392 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 profiling 384:392 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 1 89 theme immune 215:220 arg1 system 222:227 the innate immune system 204:227 the innate immune system 204:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 5 90 theme unusual 1031:1037 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 14 91 theme first 2572:2576 arg1 time 2578:2581 the first time 2568:2581 the first time 2568:2581 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 3 92 theme N-glycosylation 671:685 arg1 activation 623:632 neutrophil activation 612:632 neutrophil activation 612:632 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 3 92 theme N-glycosylation 671:685 arg1 importance 653:662 the biological importance 638:662 the biological importance of its N-glycosylation 638:685 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 8 93 theme residing 1423:1430 arg1 N-glycosylation 1400:1414 highly uniform N-glycosylation 1385:1414 highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments 1385:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 9 94 theme cell 1489:1492 arg1 mobilization 1502:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization 1469:1513 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 10 95 theme HNE 1785:1787 arg1 activity 1796:1803 HNE enzyme activity 1785:1803 HNE enzyme activity 1785:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 5 96 theme core 1039:1042 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 8 97 theme distinct 1435:1442 arg1 compartments 1455:1466 distinct neutrophil compartments 1435:1466 distinct neutrophil compartments 1435:1466 Subcellular- and site-specific glycoprofiling showed highly uniform N-glycosylation of HNE residing in distinct neutrophil compartments. 28630087 5 98 theme monoantenna 1056:1066 arg1 N-sialoglycans 1081:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans 1019:1094 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 2 99 theme individuals 555:565 arg1 granules 535:542 isolated neutrophil granules 515:542 isolated neutrophil granules of healthy individuals 515:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 1 100 theme N-glycosylated 170:183 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 100 theme N-glycosylated 170:183 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 13 101 theme isolated 2254:2261 arg1 N-glycans 2267:2275 isolated HNE N-glycans 2254:2275 isolated HNE N-glycans 2254:2275 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 6 102 contain carried 1184:1190 arg3 contrast 1146:1153 contrast 1146:1153 contrast 1146:1153 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 6 102 contain carried 1184:1190 arg2 N-glycans 1223:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 6 102 contain carried 1184:1190 arg1 Asn88 1178:1182 the mostly unoccupied Asn88 1156:1182 the mostly unoccupied Asn88 1156:1182 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 14 103 theme HNE 2529:2531 arg1 N-glycosylation 2533:2547 the unusual HNE N-glycosylation 2517:2547 the unusual HNE N-glycosylation 2517:2547 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 4 104 theme HNE 850:852 arg1 glycoprofiling 854:867 Site-specific HNE glycoprofiling 836:867 Site-specific HNE glycoprofiling 836:867 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 9 105 theme cell 1565:1568 arg1 signatures 1601:1610 cell surface-specific glycosylation signatures 1565:1610 cell surface-specific glycosylation signatures 1565:1610 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 5 106 theme occupied 1127:1134 arg1 sites 1136:1140 these two fully occupied sites 1111:1140 these two fully occupied sites 1111:1140 The equally unusual core fucosylated monoantenna complex-type N-sialoglycans also decorated these two fully occupied sites. 28630087 6 107 theme low 1257:1259 arg1 accessibility 1288:1300 low glycosylation site solvent accessibility 1257:1300 low glycosylation site solvent accessibility 1257:1300 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 2 108 gly N-glycosylation 445:459 arg1 HNE 464:466 HNE 464:466 HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals 464:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 2 109 theme heterogeneous 431:443 arg1 N-glycosylation 445:459 the heterogeneous N-glycosylation 427:459 the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals 427:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 9 110 theme glycosylation 1587:1599 arg1 signatures 1601:1610 cell surface-specific glycosylation signatures 1565:1610 cell surface-specific glycosylation signatures 1565:1610 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 10 111 theme active 1716:1721 arg1 site 1723:1726 the active site 1712:1726 the active site indicating glycan functions other than interference with HNE enzyme activity 1712:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 14 112 theme unusual 2521:2527 arg1 N-glycosylation 2533:2547 the unusual HNE N-glycosylation 2517:2547 the unusual HNE N-glycosylation 2517:2547 Taken together, these observations support that the unusual HNE N-glycosylation, here reported for the first time, is involved in modulating multiple immune functions central to inflammation and infection. 28630087 0 113 theme Spatiotemporally 37:52 arg1 Elastase 81:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 1 114 theme neutrophil 128:137 arg1 protease 192:199 an important N-glycosylated serine protease 157:199 an important N-glycosylated serine protease in the innate immune system 157:227 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 114 theme neutrophil 128:137 arg1 HNE 149:151 HNE 149:151 HNE 149:151 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 1 114 theme neutrophil 128:137 arg1 elastase 139:146 Human neutrophil elastase 122:146 Human neutrophil elastase (HNE) 122:152 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 28630087 2 115 theme neutrophil 482:491 arg1 lysates 493:499 neutrophil lysates 482:499 neutrophil lysates 482:499 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 3 116 theme surface 737:743 arg1 capture 745:751 cell surface capture 732:751 cell surface capture 732:751 The spatiotemporal expression of HNE during neutrophil activation and the biological importance of its N-glycosylation were also investigated using immunoblotting, cell surface capture, native MS, receptor interaction, protease inhibition, and bacteria growth assays. 28630087 0 117 theme Human 64:68 arg1 Elastase 81:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Spatiotemporally Regulated Human Neutrophil Elastase 37:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 9 118 theme HNE 1616:1618 arg1 regulation 1545:1554 a spatiotemporal regulation 1528:1554 a spatiotemporal regulation 1528:1554 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 6 119 theme nonfucosylated 1192:1205 arg1 N-glycans 1223:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 2 120 theme neutrophil 524:533 arg1 granules 535:542 isolated neutrophil granules 515:542 isolated neutrophil granules of healthy individuals 515:565 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 6 121 gly nonfucosylated 1192:1205 arg1 N-glycans 1223:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 nonfucosylated paucimannosidic N-glycans 1192:1231 In contrast, the mostly unoccupied Asn88 carried nonfucosylated paucimannosidic N-glycans probably resulting from low glycosylation site solvent accessibility. 28630087 0 122 theme Elastase 81:88 arg1 N-glycosylation 18:32 Paucimannose-Rich N-glycosylation 0:32 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase 0:88 Paucimannose-Rich N-glycosylation of Spatiotemporally Regulated Human Neutrophil Elastase Modulates Its Immune Functions. 28630087 9 123 theme activated 1623:1631 arg1 neutrophils 1639:1649 activated human neutrophils 1623:1649 activated human neutrophils 1623:1649 Stimulation-induced cell surface mobilization demonstrated a spatiotemporal regulation, but not cell surface-specific glycosylation signatures, of HNE in activated human neutrophils. 28630087 10 124 theme enzyme 1789:1794 arg1 activity 1796:1803 HNE enzyme activity 1785:1803 HNE enzyme activity 1785:1803 The three glycosylation sites of HNE were located distal to the active site indicating glycan functions other than interference with HNE enzyme activity. 28630087 7 125 gly glycosylated 1318:1329 arg1 Asn185 1303:1308 Asn185 1303:1308 Asn185 1303:1308 Asn185 was not glycosylated. 28630087 11 126 theme preferential 1865:1876 arg1 binding 1878:1884 preferential binding 1865:1884 preferential binding to human mannose binding lectin 1865:1916 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 2 127 theme glycoprotein 371:382 arg1 profiling 384:392 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 profiling 384:392 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 12 128 theme glycoform-glycoform 2168:2186 arg1 interactions 2188:2199 preferred glycoform-glycoform interactions 2158:2199 preferred glycoform-glycoform interactions 2158:2199 The heavily N-glycosylated HNE protease inhibitor, α1-antitrypsin, displayed concentration-dependent complex formation and preferred glycoform-glycoform interactions with HNE. 28630087 13 129 theme bacteriostatic 2410:2423 arg1 activity 2425:2432 some bacteriostatic activity 2405:2432 some bacteriostatic activity 2405:2432 Finally, both enzymatically active HNE and isolated HNE N-glycans demonstrated low micromolar concentration-dependent growth inhibition of clinically-relevant Pseudomonas aeruginosa, suggesting some bacteriostatic activity is conferred by the HNE N-glycans. 28630087 11 130 theme complement 2012:2021 arg1 activation 2023:2032 complement activation 2012:2032 complement activation 2012:2032 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 4 131 theme unusual 887:893 arg1 N-glycans 911:919 unusual paucimannosidic N-glycans 887:919 unusual paucimannosidic N-glycans 887:919 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 4 131 theme unusual 887:893 arg1 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Manα1,6Manβ1,4GlcNAcβ1,4 935:958 Site-specific HNE glycoprofiling demonstrated that unusual paucimannosidic N-glycans, particularly Manα1,6Manβ1,4GlcNAcβ1,4(Fucα1,6)GlcNAcβ, predominantly occupied Asn124 and Asn173. 28630087 2 132 theme glycopeptide 354:365 arg1 profiling 384:392 LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling 331:392 profiling 384:392 Herein, LC-MS/MS-based glycan, glycopeptide and glycoprotein profiling were utilized to first determine the heterogeneous N-glycosylation of HNE purified from neutrophil lysates and then from isolated neutrophil granules of healthy individuals. 28630087 11 133 theme HNE 1936:1938 arg1 sialoglycoforms 1940:1954 the HNE sialoglycoforms 1932:1954 the HNE sialoglycoforms 1932:1954 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 11 134 theme mannose 1895:1901 arg1 lectin 1911:1916 human mannose binding lectin 1889:1916 human mannose binding lectin 1889:1916 Functionally, the paucimannosidic HNE glycoforms displayed preferential binding to human mannose binding lectin compared with the HNE sialoglycoforms, suggesting a glycoform-dependent involvement of HNE in complement activation. 28630087 1 135 theme immune-modulating 252:268 arg1 functions 270:278 immune-modulating functions 252:278 immune-modulating functions 252:278 Human neutrophil elastase (HNE) is an important N-glycosylated serine protease in the innate immune system, but the structure and immune-modulating functions of HNE N-glycosylation remain undescribed. 26784534 8 0 theme A1 976:977 arg1 domain 979:984 the A1 domain 972:984 the A1 domain 972:984 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 8 1 gly glycopeptides 949:961 arg2 glycopeptides 949:961 the glycopeptides 945:961 the glycopeptides flanking the A1 domain 945:984 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 11 2 theme further 1687:1693 arg1 studies 1695:1701 further studies 1687:1701 further studies exploring the functional and structural implications of O-glycosylation in VWF 1687:1780 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 1 3 theme putative 225:232 arg1 sites 250:254 the 10 putative O-glycosylation sites 218:254 the 10 putative O-glycosylation sites 218:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 4 theme sites 250:254 arg1 occupancy 205:213 the occupancy 201:213 the occupancy of the 10 putative O-glycosylation sites 201:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 6 5 from sites 740:744 arg1 majority 685:692 The majority 681:692 The majority of the glycan structures on all glycosylation sites 681:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 9 6 attach present 1277:1283 arg1 glycopeptides 1293:1305 both glycopeptides 1288:1305 both glycopeptides flanking the A1 domain 1288:1328 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 6 attach present 1277:1283 arg2 motifs 1265:1270 Disialosyl motifs 1254:1270 Disialosyl motifs 1254:1270 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 6 attach present 1277:1283 arg1 site 1355:1358 the glycosylation site T(2298) 1337:1366 the glycosylation site T(2298) in the C1 domain 1337:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 8 7 theme core-2-type 1015:1025 arg1 O-glycan 1027:1034 generally more core-2-type O-glycan 1000:1034 generally more core-2-type O-glycan 1000:1034 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 10 8 theme core 1424:1427 arg1 O-glycans 1431:1439 core 2 O-glycans 1424:1439 core 2 O-glycans 1424:1439 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 11 9 theme O-glycosylation 1615:1629 arg1 sites 1631:1635 all 10 O-glycosylation sites 1608:1635 all 10 O-glycosylation sites 1608:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 9 10 theme A1 1320:1321 arg1 domain 1323:1328 the A1 domain 1316:1328 the A1 domain 1316:1328 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 11 from site 1355:1358 arg1 domain 1378:1383 the C1 domain 1371:1383 the C1 domain 1371:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 6 12 from structures 708:717 arg1 sites 740:744 all glycosylation sites 722:744 all glycosylation sites 722:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 9 13 gly glycopeptides 1293:1305 arg2 glycopeptides 1293:1305 both glycopeptides 1288:1305 both glycopeptides flanking the A1 domain 1288:1328 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 8 14 gly glycopeptide 1184:1195 arg2 glycopeptide 1184:1195 the N-terminal Cluster 1 glycopeptide 1159:1195 the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ) 1159:1251 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 1 15 from information 186:196 arg1 occupancy 205:213 the occupancy 201:213 the occupancy of the 10 putative O-glycosylation sites 201:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 8 16 theme N-terminal 1163:1172 arg1 glycopeptide 1184:1195 the N-terminal Cluster 1 glycopeptide 1159:1195 the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ) 1159:1251 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 6 17 from majority 685:692 arg1 sites 740:744 all glycosylation sites 722:744 all glycosylation sites 722:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 10 18 theme O-glycans 1431:1439 arg1 sulfation 1411:1419 sulfation 1411:1419 sulfation of core 2 O-glycans 1411:1439 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 10 18 theme O-glycans 1431:1439 arg1 presence 1449:1456 the presence 1445:1456 the presence of the rare Tn antigen 1445:1479 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 11 19 theme semi-quantitative 1551:1567 arg1 distribution 1569:1580 the qualitative and semi-quantitative distribution 1531:1580 the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites 1531:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 20 theme O-glycosylation 1759:1773 arg1 implications 1743:1754 the functional and structural implications 1713:1754 the functional and structural implications of O-glycosylation 1713:1773 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 21 theme qualitative 1535:1545 arg1 distribution 1569:1580 the qualitative and semi-quantitative distribution 1531:1580 the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites 1531:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 4 22 theme purified 513:520 arg1 glycopeptides 522:534 the purified glycopeptides 509:534 the purified glycopeptides 509:534 Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry. 26784534 8 23 gly glycopeptide 1076:1087 arg2 glycopeptide 1076:1087 the C-terminal Cluster 2 glycopeptide 1051:1087 the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) 1051:1143 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 11 24 theme structures 1594:1603 arg1 distribution 1569:1580 the qualitative and semi-quantitative distribution 1531:1580 the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites 1531:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 1 25 gly O-glycosylation 77:91 arg1 factor 111:116 von Willebrand factor 96:116 von Willebrand factor (VWF) 96:122 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 25 gly O-glycosylation 77:91 arg1 VWF 119:121 VWF 119:121 VWF 119:121 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 25 gly O-glycosylation 77:91 arg1 functions 144:152 its functions 140:152 its functions 140:152 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 26 gly occupancy 205:213 arg2 sites 250:254 the 10 putative O-glycosylation sites 218:254 the 10 putative O-glycosylation sites 218:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 8 27 theme C-terminal 1055:1064 arg1 glycopeptide 1076:1087 the C-terminal Cluster 2 glycopeptide 1051:1087 the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) 1051:1143 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 5 28 theme predicted 628:636 arg1 sites 654:658 all 10 predicted O-glycosylation sites 621:658 all 10 predicted O-glycosylation sites in VWF 621:665 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 8 29 attach present 1040:1046 arg1 glycopeptide 1076:1087 the C-terminal Cluster 2 glycopeptide 1051:1087 the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) 1051:1143 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 8 29 attach present 1040:1046 arg2 O-glycan 1027:1034 generally more core-2-type O-glycan 1000:1034 generally more core-2-type O-glycan 1000:1034 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 6 30 theme glycosylation 726:738 arg1 sites 740:744 all glycosylation sites 722:744 all glycosylation sites 722:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 10 31 theme Tn 1470:1471 arg1 antigen 1473:1479 the rare Tn antigen 1461:1479 the rare Tn antigen 1461:1479 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 3 32 theme Tryptic 355:361 arg1 VWF-O-glycopeptides 363:381 Tryptic VWF-O-glycopeptides 355:381 Tryptic VWF-O-glycopeptides 355:381 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 3 33 theme affinity 407:414 arg1 chromatography 416:429 lectin affinity chromatography 400:429 lectin affinity chromatography 400:429 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 0 34 theme Site-specific 0:12 arg1 analysis 14:21 Site-specific analysis 0:21 Site-specific analysis of von Willebrand factor O-glycosylation 0:62 Site-specific analysis of von Willebrand factor O-glycosylation. 26784534 1 35 theme Willebrand 100:109 arg1 VWF 119:121 VWF 119:121 VWF 119:121 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 35 theme Willebrand 100:109 arg1 factor 111:116 von Willebrand factor 96:116 von Willebrand factor (VWF) 96:122 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 4 36 theme mass 579:582 arg1 spectrometry 584:595 mass spectrometry 579:595 mass spectrometry 579:595 Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry. 26784534 6 37 theme glycan 701:706 arg1 structures 708:717 the glycan structures 697:717 the glycan structures on all glycosylation sites 697:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 3 38 theme liquid 472:477 arg1 chromatography 479:492 reverse-phase high-performance liquid chromatography 441:492 reverse-phase high-performance liquid chromatography 441:492 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 5 39 from sites 654:658 arg1 VWF 663:665 VWF 663:665 VWF 663:665 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 11 40 theme first 1507:1511 arg1 This 1495:1498 This 1495:1498 This 1495:1498 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 40 theme first 1507:1511 arg1 CONCLUSIONS 1482:1492 CONCLUSIONS 1482:1492 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 1482:1781 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 40 theme first 1507:1511 arg1 study 1513:1517 the first study 1503:1517 the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites 1503:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 3 41 theme reverse-phase 441:453 arg1 chromatography 479:492 reverse-phase high-performance liquid chromatography 441:492 reverse-phase high-performance liquid chromatography 441:492 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 0 42 theme Willebrand 30:39 arg1 factor 41:46 von Willebrand factor 26:46 von Willebrand factor O-glycosylation 26:62 Site-specific analysis of von Willebrand factor O-glycosylation. 26784534 11 43 theme starting 1668:1675 arg1 point 1677:1681 a valuable starting point 1657:1681 a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF 1657:1780 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 0 44 theme O-glycosylation 48:62 arg1 analysis 14:21 Site-specific analysis 0:21 Site-specific analysis of von Willebrand factor O-glycosylation 0:62 Site-specific analysis of von Willebrand factor O-glycosylation. 26784534 6 45 gly glycosylation 726:738 arg2 sites 740:744 all glycosylation sites 722:744 all glycosylation sites 722:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 8 46 theme glycopeptides 949:961 arg1 Analysis 933:940 Analysis 933:940 Analysis of the glycopeptides flanking the A1 domain 933:984 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 6 47 theme core 773:776 arg1 O-glycan 780:787 disialyl core 1 O-glycan 764:787 disialyl core 1 O-glycan 764:787 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 10 48 theme antigen 1473:1479 arg1 sulfation 1411:1419 sulfation 1411:1419 sulfation of core 2 O-glycans 1411:1439 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 10 48 theme antigen 1473:1479 arg1 presence 1449:1456 the presence 1445:1456 the presence of the rare Tn antigen 1445:1479 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 7 49 theme glycosylation 912:924 arg1 sites 926:930 all 10 glycosylation sites 905:930 all 10 glycosylation sites 905:930 The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. 26784534 9 50 theme Disialosyl 1254:1263 arg1 motifs 1265:1270 Disialosyl motifs 1254:1270 Disialosyl motifs 1254:1270 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 2 51 theme VWF 325:327 arg1 O-glycosylation 329:343 VWF O-glycosylation 325:343 VWF O-glycosylation 325:343 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 11 52 dep study 1513:1517 arg1 provide 1649:1655 provide 1649:1655 will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF 1644:1780 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 9 53 gly glycosylation 1341:1353 arg2 site 1355:1358 the glycosylation site T(2298) 1337:1366 the glycosylation site T(2298) in the C1 domain 1337:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 1 54 theme O-glycosylation 234:248 arg1 sites 250:254 the 10 putative O-glycosylation sites 218:254 the 10 putative O-glycosylation sites 218:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 8 55 located present 1040:1046 arg1 glycopeptide 1076:1087 the C-terminal Cluster 2 glycopeptide 1051:1087 the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) 1051:1143 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 8 55 located present 1040:1046 arg2 O-glycan 1027:1034 generally more core-2-type O-glycan 1000:1034 generally more core-2-type O-glycan 1000:1034 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 4 56 gly glycopeptides 522:534 arg2 glycopeptides 522:534 the purified glycopeptides 509:534 the purified glycopeptides 509:534 Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry. 26784534 5 57 used occupied 671:678 arg2 sites 654:658 all 10 predicted O-glycosylation sites 621:658 all 10 predicted O-glycosylation sites in VWF 621:665 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 8 58 theme Cluster 1174:1180 arg1 glycopeptide 1184:1195 the N-terminal Cluster 1 glycopeptide 1159:1195 the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ) 1159:1251 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 2 59 theme study 285:289 arg1 analysis 313:320 the site-specific analysis 295:320 the site-specific analysis of VWF O-glycosylation 295:343 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 59 theme study 285:289 arg1 OBJECTIVES 257:266 OBJECTIVES 257:266 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 257:344 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 59 theme study 285:289 arg1 aim 273:275 The aim 269:275 The aim of this study 269:289 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 9 60 theme glycosylation 1341:1353 arg1 site 1355:1358 the glycosylation site T(2298) 1337:1366 the glycosylation site T(2298) in the C1 domain 1337:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 11 61 from distribution 1569:1580 arg1 sites 1631:1635 all 10 O-glycosylation sites 1608:1635 all 10 O-glycosylation sites 1608:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 7 62 gly glycosylation 912:924 arg2 sites 926:930 all 10 glycosylation sites 905:930 all 10 glycosylation sites 905:930 The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. 26784534 7 62 gly glycosylation 912:924 arg2 10 909:910 10 909:910 10 909:910 The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. 26784534 1 63 theme functions 144:152 arg1 many 132:135 many 132:135 many 132:135 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 63 theme functions 144:152 arg1 functions 144:152 its functions 140:152 its functions 140:152 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 8 64 theme Cluster 1066:1072 arg1 glycopeptide 1076:1087 the C-terminal Cluster 2 glycopeptide 1051:1087 the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) 1051:1143 Analysis of the glycopeptides flanking the A1 domain revealed that generally more core-2-type O-glycan was present on the C-terminal Cluster 2 glycopeptide (encompassing T(1468) , T(1477) , S(1486) and T(1487) ) compared with the N-terminal Cluster 1 glycopeptide (encompassing T(1248) , T(1255) , T(1256) and S(1263) ). 26784534 9 65 theme C1 1375:1376 arg1 domain 1378:1383 the C1 domain 1371:1383 the C1 domain 1371:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 11 66 theme O-glycan 1585:1592 arg1 structures 1594:1603 O-glycan structures 1585:1603 O-glycan structures 1585:1603 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 3 67 gly VWF-O-glycopeptides 363:381 arg2 VWF-O-glycopeptides 363:381 Tryptic VWF-O-glycopeptides 355:381 Tryptic VWF-O-glycopeptides 355:381 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 5 68 theme O-glycosylation 638:652 arg1 sites 654:658 all 10 predicted O-glycosylation sites 621:658 all 10 predicted O-glycosylation sites in VWF 621:665 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 9 69 located present 1277:1283 arg1 glycopeptides 1293:1305 both glycopeptides 1288:1305 both glycopeptides flanking the A1 domain 1288:1328 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 69 located present 1277:1283 arg2 motifs 1265:1270 Disialosyl motifs 1254:1270 Disialosyl motifs 1254:1270 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 70 dep site 1355:1358 arg1 T 1360:1360 T 1360:1360 the glycosylation site T(2298) in the C1 domain 1337:1383 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 9 70 dep site 1355:1358 arg1 2298 1362:1365 2298 1362:1365 2298 1362:1365 Disialosyl motifs were present on both glycopeptides flanking the A1 domain and on the glycosylation site T(2298) in the C1 domain. 26784534 10 71 theme rare 1465:1468 arg1 antigen 1473:1479 the rare Tn antigen 1461:1479 the rare Tn antigen 1461:1479 In addition, we identify sulfation of core 2 O-glycans and the presence of the rare Tn antigen. 26784534 1 72 theme von 96:98 arg1 VWF 119:121 VWF 119:121 VWF 119:121 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 72 theme von 96:98 arg1 factor 111:116 von Willebrand factor 96:116 von Willebrand factor (VWF) 96:122 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 6 73 theme structures 708:717 arg1 majority 685:692 The majority 681:692 The majority of the glycan structures on all glycosylation sites 681:744 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 11 74 theme functional 1717:1726 arg1 implications 1743:1754 the functional and structural implications 1713:1754 the functional and structural implications of O-glycosylation 1713:1773 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 0 75 theme von 26:28 arg1 factor 41:46 von Willebrand factor 26:46 von Willebrand factor O-glycosylation 26:62 Site-specific analysis of von Willebrand factor O-glycosylation. 26784534 1 76 gly O-glycosylation 234:248 arg2 sites 250:254 the 10 putative O-glycosylation sites 218:254 the 10 putative O-glycosylation sites 218:254 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 76 gly O-glycosylation 234:248 arg2 10 222:223 10 222:223 10 222:223 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 1 77 theme factor 111:116 arg1 O-glycosylation 77:91 O-glycosylation 77:91 O-glycosylation of von Willebrand factor (VWF) 77:122 BACKGROUND: O-glycosylation of von Willebrand factor (VWF) affects many of its functions; however, there is currently no information on the occupancy of the 10 putative O-glycosylation sites. 26784534 0 78 theme factor 41:46 arg1 O-glycosylation 48:62 von Willebrand factor O-glycosylation 26:62 von Willebrand factor O-glycosylation 26:62 Site-specific analysis of von Willebrand factor O-glycosylation. 26784534 7 79 theme O-glycan 813:820 arg1 presence 794:801 The presence 790:801 The presence of core 2 O-glycan 790:820 The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. 26784534 3 80 theme lectin 400:405 arg1 chromatography 416:429 lectin affinity chromatography 400:429 lectin affinity chromatography 400:429 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 4 81 theme glycosidase 553:563 arg1 digestion 565:573 glycosidase digestion 553:573 glycosidase digestion 553:573 Subsequently, the purified glycopeptides were analyzed by glycosidase digestion and mass spectrometry. 26784534 3 82 theme high-performance 455:470 arg1 chromatography 479:492 reverse-phase high-performance liquid chromatography 441:492 reverse-phase high-performance liquid chromatography 441:492 METHODS: Tryptic VWF-O-glycopeptides were isolated by lectin affinity chromatography and/or by reverse-phase high-performance liquid chromatography. 26784534 7 83 theme core 806:809 arg1 O-glycan 813:820 core 2 O-glycan 806:820 core 2 O-glycan 806:820 The presence of core 2 O-glycan was also confirmed; interestingly, this structure was not evenly distributed among all 10 glycosylation sites. 26784534 2 84 theme site-specific 299:311 arg1 analysis 313:320 the site-specific analysis 295:320 the site-specific analysis of VWF O-glycosylation 295:343 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 84 theme site-specific 299:311 arg1 OBJECTIVES 257:266 OBJECTIVES 257:266 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 257:344 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 84 theme site-specific 299:311 arg1 aim 273:275 The aim 269:275 The aim of this study 269:289 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 11 85 gly O-glycosylation 1615:1629 arg2 sites 1631:1635 all 10 O-glycosylation sites 1608:1635 all 10 O-glycosylation sites 1608:1635 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 85 gly O-glycosylation 1615:1629 arg2 10 1612:1613 10 1612:1613 10 1612:1613 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 6 86 theme disialyl 764:771 arg1 O-glycan 780:787 disialyl core 1 O-glycan 764:787 disialyl core 1 O-glycan 764:787 The majority of the glycan structures on all glycosylation sites is represented by disialyl core 1 O-glycan. 26784534 11 87 theme valuable 1659:1666 arg1 point 1677:1681 a valuable starting point 1657:1681 a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF 1657:1780 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 11 88 theme structural 1732:1741 arg1 implications 1743:1754 the functional and structural implications 1713:1754 the functional and structural implications of O-glycosylation 1713:1773 CONCLUSIONS: This is the first study to describe the qualitative and semi-quantitative distribution of O-glycan structures on all 10 O-glycosylation sites, which will provide a valuable starting point for further studies exploring the functional and structural implications of O-glycosylation in VWF. 26784534 5 89 gly O-glycosylation 638:652 arg2 sites 654:658 all 10 predicted O-glycosylation sites 621:658 all 10 predicted O-glycosylation sites in VWF 621:665 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 5 89 gly O-glycosylation 638:652 arg2 10 625:626 10 625:626 10 625:626 RESULTS: We found that all 10 predicted O-glycosylation sites in VWF are occupied. 26784534 2 90 theme O-glycosylation 329:343 arg1 analysis 313:320 the site-specific analysis 295:320 the site-specific analysis of VWF O-glycosylation 295:343 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 90 theme O-glycosylation 329:343 arg1 OBJECTIVES 257:266 OBJECTIVES 257:266 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 257:344 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26784534 2 90 theme O-glycosylation 329:343 arg1 aim 273:275 The aim 269:275 The aim of this study 269:289 OBJECTIVES: The aim of this study was the site-specific analysis of VWF O-glycosylation. 26189796 0 0 theme gastric 92:98 arg1 cancer 100:105 gastric cancer 92:105 gastric cancer 92:105 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 5 1 theme junctions 1190:1198 arg1 stability 1164:1172 stability 1164:1172 stability 1164:1172 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 1 theme junctions 1190:1198 arg1 localization 1106:1117 E-cadherin cellular localization 1086:1117 E-cadherin cellular localization 1086:1117 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 1 theme junctions 1190:1198 arg1 formation 1130:1138 cis-dimer formation 1120:1138 cis-dimer formation 1120:1138 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 1 theme junctions 1190:1198 arg1 assembly 1151:1158 molecular assembly 1141:1158 molecular assembly 1141:1158 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 2 2 theme site-specific 369:381 arg1 glycosylation 383:395 site-specific glycosylation 369:395 site-specific glycosylation of E-cadherin 369:409 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 5 3 theme gastric 1263:1269 arg1 carcinomas 1271:1280 human gastric carcinomas 1257:1280 human gastric carcinomas 1257:1280 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 6 4 theme tumor 1570:1574 arg1 suppression 1576:1586 tumor suppression 1570:1586 tumor suppression 1570:1586 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 5 5 theme molecular 1141:1149 arg1 assembly 1151:1158 molecular assembly 1141:1158 molecular assembly 1141:1158 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 6 from modification 939:950 arg1 site 980:983 this specific asparagine site 955:983 this specific asparagine site 955:983 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 1 7 theme gastric 159:165 arg1 carcinogenesis 167:180 gastric carcinogenesis 159:180 gastric carcinogenesis 159:180 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 1 8 theme deleterious 273:283 arg1 effect 285:290 a deleterious effect 271:290 a deleterious effect 271:290 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 3 9 from expression 673:682 arg1 mucosa 699:704 the gastric mucosa 687:704 the gastric mucosa 687:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 4 10 dep In 707:708 arg1 vitro 710:714 vitro 710:714 vitro 710:714 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 5 11 located observed 1245:1252 arg2 aggregation 1214:1224 cell-cell aggregation 1204:1224 cell-cell aggregation 1204:1224 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 11 located observed 1245:1252 arg2 junctions 1190:1198 adherens junctions 1181:1198 adherens junctions 1181:1198 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 11 located observed 1245:1252 arg1 carcinomas 1271:1280 human gastric carcinomas 1257:1280 human gastric carcinomas 1257:1280 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 4 12 gly N-glycosylation 772:786 arg2 four 757:760 four 757:760 four 757:760 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 12 gly N-glycosylation 772:786 arg2 E-cadherin 797:806 E-cadherin 797:806 E-cadherin 797:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 12 gly N-glycosylation 772:786 arg1 E-cadherin 797:806 E-cadherin 797:806 E-cadherin 797:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 12 gly N-glycosylation 772:786 arg2 sites 788:792 the four potential N-glycosylation sites 753:792 the four potential N-glycosylation sites of E-cadherin 753:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 13 theme E-cadherin 797:806 arg1 sites 788:792 the four potential N-glycosylation sites 753:792 the four potential N-glycosylation sites of E-cadherin 753:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 13 theme E-cadherin 797:806 arg1 E-cadherin 797:806 E-cadherin 797:806 E-cadherin 797:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 3 14 theme N-acetylglucosaminyltransferase 567:597 arg1 GnT-V 602:606 GnT-V 602:606 GnT-V 602:606 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 3 14 theme N-acetylglucosaminyltransferase 567:597 arg1 V 599:599 N-acetylglucosaminyltransferase V 567:599 N-acetylglucosaminyltransferase V (GnT-V) 567:607 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 6 15 from effect 1488:1493 arg1 E-cadherin 1498:1507 E-cadherin 1498:1507 E-cadherin 1498:1507 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 4 16 mod modified 853:860 arg1 site 828:831 the key site 820:831 the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V 820:915 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 16 mod modified 853:860 arg3 N-glycans 888:896 β1,6 GlcNAc-branched N-glycans 867:896 β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V 867:915 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 16 mod modified 853:860 arg1 Asn-554 809:815 Asn-554 809:815 Asn-554 809:815 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 17 theme GlcNAc-branched 872:886 arg1 N-glycans 888:896 β1,6 GlcNAc-branched N-glycans 867:896 β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V 867:915 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 3 18 from pattern 651:657 arg1 mucosa 699:704 the gastric mucosa 687:704 the gastric mucosa 687:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 1 19 theme carcinogenesis 167:180 arg1 process 148:154 the process 144:154 the process of gastric carcinogenesis 144:180 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 6 20 theme protective 1477:1486 arg1 effect 1488:1493 a protective effect 1475:1493 a protective effect on E-cadherin 1475:1507 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 4 21 theme β1,6 867:870 arg1 N-glycans 888:896 β1,6 GlcNAc-branched N-glycans 867:896 β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V 867:915 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 1 22 from molecule 132:139 arg1 process 148:154 the process 144:154 the process of gastric carcinogenesis 144:180 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 5 23 theme critical 1030:1037 arg1 functions 1039:1047 its critical functions 1026:1047 its critical functions in gastric cancer cells 1026:1071 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 24 from functions 1039:1047 arg1 cells 1067:1071 gastric cancer cells 1052:1071 gastric cancer cells 1052:1071 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 0 25 theme E-cadherin 11:20 arg1 N-glycosylation 31:45 E-cadherin aberrant N-glycosylation 11:45 E-cadherin aberrant N-glycosylation 11:45 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 5 26 theme human 1257:1261 arg1 carcinomas 1271:1280 human gastric carcinomas 1257:1280 human gastric carcinomas 1257:1280 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 4 27 theme N-glycosylation 772:786 arg1 sites 788:792 the four potential N-glycosylation sites 753:792 the four potential N-glycosylation sites of E-cadherin 753:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 27 theme N-glycosylation 772:786 arg1 E-cadherin 797:806 E-cadherin 797:806 E-cadherin 797:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 1 28 theme cell 295:298 arg1 adhesion 300:307 cell adhesion 295:307 cell adhesion associated with tumor cell invasion 295:343 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 4 29 theme potential 762:770 arg1 sites 788:792 the four potential N-glycosylation sites 753:792 the four potential N-glycosylation sites of E-cadherin 753:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 29 theme potential 762:770 arg1 E-cadherin 797:806 E-cadherin 797:806 E-cadherin 797:806 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 1 30 theme posttranslational 190:206 arg1 modifications 208:220 its posttranslational modifications 186:220 its posttranslational modifications by N-glycosylation 186:239 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 5 31 theme gastric 1052:1058 arg1 cells 1067:1071 gastric cancer cells 1052:1071 gastric cancer cells 1052:1071 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 2 32 gly glycosylation 383:395 arg1 E-cadherin 400:409 E-cadherin 400:409 E-cadherin 400:409 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 5 33 theme E-cadherin 1086:1095 arg1 localization 1106:1117 E-cadherin cellular localization 1086:1117 E-cadherin cellular localization 1086:1117 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 6 34 theme site-specific 1316:1328 arg1 glycosylation 1330:1342 this site-specific glycosylation 1311:1342 this site-specific glycosylation 1311:1342 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 2 35 theme gastric 444:450 arg1 cells 459:463 gastric cancer cells 444:463 gastric cancer cells 444:463 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 2 36 theme cancer 452:457 arg1 cells 459:463 gastric cancer cells 444:463 gastric cancer cells 444:463 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 5 37 theme E-cadherin 988:997 arg1 modification 939:950 This aberrant glycan modification 918:950 This aberrant glycan modification on this specific asparagine site of E-cadherin 918:997 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 3 38 theme clinical 528:535 arg1 samples 537:543 human clinical samples 522:543 human clinical samples 522:543 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 3 39 theme expression 673:682 arg1 pattern 651:657 an abnormal pattern 639:657 an abnormal pattern of E-cadherin expression in the gastric mucosa 639:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 3 40 theme human 522:526 arg1 samples 537:543 human clinical samples 522:543 human clinical samples 522:543 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 5 41 theme specific 960:967 arg1 site 980:983 this specific asparagine site 955:983 this specific asparagine site 955:983 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 3 42 theme gastric 691:697 arg1 mucosa 699:704 the gastric mucosa 687:704 the gastric mucosa 687:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 6 43 theme functional 1525:1534 arg1 dysregulation 1536:1548 its functional dysregulation 1521:1548 its functional dysregulation 1521:1548 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 2 44 theme E-cadherin 400:409 arg1 glycosylation 383:395 site-specific glycosylation 369:395 site-specific glycosylation of E-cadherin 369:409 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 1 45 theme tumor 325:329 arg1 invasion 336:343 tumor cell invasion 325:343 tumor cell invasion 325:343 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 0 46 from function 80:87 arg1 cancer 100:105 gastric cancer 92:105 gastric cancer 92:105 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 2 47 theme defective 422:430 arg1 function 432:439 its defective function 418:439 its defective function in gastric cancer cells 418:463 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 1 48 theme cell 331:334 arg1 invasion 336:343 tumor cell invasion 325:343 tumor cell invasion 325:343 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 3 49 theme -mediated 608:616 arg1 glycosylation 618:630 N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation 567:630 N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation 567:630 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 5 50 theme cell-cell 1204:1212 arg1 aggregation 1214:1224 cell-cell aggregation 1204:1224 cell-cell aggregation 1204:1224 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 51 theme cellular 1097:1104 arg1 localization 1106:1117 E-cadherin cellular localization 1086:1117 E-cadherin cellular localization 1086:1117 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 0 52 gly N-glycosylation 31:45 arg1 Asn-554 50:56 Asn-554 50:56 Asn-554 50:56 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 5 53 theme aggregation 1214:1224 arg1 stability 1164:1172 stability 1164:1172 stability 1164:1172 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 53 theme aggregation 1214:1224 arg1 localization 1106:1117 E-cadherin cellular localization 1086:1117 E-cadherin cellular localization 1086:1117 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 53 theme aggregation 1214:1224 arg1 formation 1130:1138 cis-dimer formation 1120:1138 cis-dimer formation 1120:1138 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 53 theme aggregation 1214:1224 arg1 assembly 1151:1158 molecular assembly 1141:1158 molecular assembly 1141:1158 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 1 54 theme central 124:130 arg1 molecule 132:139 a central molecule 122:139 a central molecule in the process of gastric carcinogenesis 122:180 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 1 54 theme central 124:130 arg1 E-cadherin 108:117 E-cadherin 108:117 E-cadherin 108:117 E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. 26189796 5 55 theme cancer 1060:1065 arg1 cells 1067:1071 gastric cancer cells 1052:1071 gastric cancer cells 1052:1071 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 6 56 theme branched 1398:1405 arg1 structures 1407:1416 the deleterious branched structures 1382:1416 the deleterious branched structures 1382:1416 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 4 57 theme In 707:708 arg1 models 716:721 In vitro models 707:721 In vitro models 707:721 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 58 theme key 824:826 arg1 site 828:831 the key site 820:831 the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V 820:915 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 4 58 theme key 824:826 arg1 Asn-554 809:815 Asn-554 809:815 Asn-554 809:815 In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. 26189796 3 59 theme transgenic 495:504 arg1 models 511:516 transgenic mice models 495:516 transgenic mice models 495:516 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 6 60 theme deleterious 1386:1396 arg1 structures 1407:1416 the deleterious branched structures 1382:1416 the deleterious branched structures 1382:1416 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 26189796 3 61 theme E-cadherin 662:671 arg1 expression 673:682 E-cadherin expression 662:682 E-cadherin expression in the gastric mucosa 662:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 2 62 from function 432:439 arg1 cells 459:463 gastric cancer cells 444:463 gastric cancer cells 444:463 However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. 26189796 3 63 theme mice 506:509 arg1 models 511:516 transgenic mice models 495:516 transgenic mice models 495:516 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 5 64 theme asparagine 969:978 arg1 site 980:983 this specific asparagine site 955:983 this specific asparagine site 955:983 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 3 65 from mucosa 699:704 arg1 pattern 651:657 an abnormal pattern 639:657 an abnormal pattern of E-cadherin expression in the gastric mucosa 639:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 5 66 theme cis-dimer 1120:1128 arg1 formation 1130:1138 cis-dimer formation 1120:1138 cis-dimer formation 1120:1138 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 3 67 theme abnormal 642:649 arg1 pattern 651:657 an abnormal pattern 639:657 an abnormal pattern of E-cadherin expression in the gastric mucosa 639:704 Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. 26189796 5 68 theme aberrant 923:930 arg1 modification 939:950 This aberrant glycan modification 918:950 This aberrant glycan modification on this specific asparagine site of E-cadherin 918:997 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 5 69 dep junctions 1190:1198 arg1 the 1177:1179 the 1177:1179 the 1177:1179 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 0 70 theme critical 71:78 arg1 function 80:87 its critical function 67:87 its critical function in gastric cancer 67:105 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 5 71 theme adherens 1181:1188 arg1 junctions 1190:1198 adherens junctions 1181:1198 adherens junctions 1181:1198 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 0 72 theme aberrant 22:29 arg1 N-glycosylation 31:45 E-cadherin aberrant N-glycosylation 11:45 E-cadherin aberrant N-glycosylation 11:45 Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. 26189796 5 73 theme glycan 932:937 arg1 modification 939:950 This aberrant glycan modification 918:950 This aberrant glycan modification on this specific asparagine site of E-cadherin 918:997 This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. 26189796 6 74 theme silencing 1446:1454 arg1 GnT-V 1456:1460 silencing GnT-V 1446:1460 silencing GnT-V 1446:1460 Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression. 27023912 5 0 theme glycan 786:791 arg1 chains 793:798 the N-linked glycan chains 773:798 the N-linked glycan chains 773:798 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 2 1 theme enzyme 375:380 arg1 development 337:347 the development 333:347 the development of a catalytically active enzyme 333:380 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 0 2 theme human 69:73 arg1 glucocerebrosidase 75:92 human glucocerebrosidase 69:92 human glucocerebrosidase 69:92 Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. 27023912 2 3 theme active 368:373 arg1 enzyme 375:380 a catalytically active enzyme 352:380 a catalytically active enzyme 352:380 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 5 4 link N-linked 777:784 arg1 chains 793:798 the N-linked glycan chains 773:798 the N-linked glycan chains 773:798 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 7 5 theme experimental 1218:1229 arg1 observation 1231:1241 the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19 1214:1313 the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19 1214:1313 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 3 6 theme glycosylation 542:554 arg1 basis 461:465 the molecular basis 447:465 the molecular basis of the relevance of N-glycosylation over GCase activity 447:521 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 3 6 theme glycosylation 542:554 arg1 effects 531:537 the effects 527:537 the effects of glycosylation over its structure and dynamics 527:586 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 4 7 theme molecular 728:736 arg1 simulations 747:757 triplicate unbiased molecular dynamics simulations 708:757 triplicate unbiased molecular dynamics simulations 708:757 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 7 8 theme catalytic 1088:1096 arg1 dyad 1098:1101 an increasing catalytic dyad 1074:1101 an increasing catalytic dyad organization 1074:1114 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 6 9 theme mutations 1044:1052 arg1 mutations 1044:1052 the most prevalent GCase mutations 1019:1052 the most prevalent GCase mutations 1019:1052 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 6 9 theme mutations 1044:1052 arg1 one 1012:1014 one 1012:1014 one 1012:1014 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 4 10 theme GCase 653:657 arg1 enzyme 659:664 GCase enzyme 653:664 GCase enzyme 653:664 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 5 11 theme stabilization 828:840 arg1 effects 842:848 local conformational stabilization effects 807:848 local conformational stabilization effects 807:848 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 7 12 theme dyad 1098:1101 arg1 organization 1103:1114 an increasing catalytic dyad organization 1074:1114 an increasing catalytic dyad organization 1074:1114 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 2 13 theme first 403:407 arg1 site 409:412 the first site 399:412 the first site 399:412 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 7 14 theme first 1179:1183 arg1 explanation 1198:1208 the first atomic-level explanation 1175:1208 the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19 1175:1313 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 1 15 gly glycoprotein 242:253 arg1 glycoprotein 242:253 a peripheral membrane-associated glycoprotein 209:253 a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 209:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 15 gly glycoprotein 242:253 arg1 enzyme 201:206 glucocerebrosidase (GCase) enzyme 174:206 glucocerebrosidase (GCase) enzyme 174:206 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 7 16 theme atomic-level 1185:1196 arg1 explanation 1198:1208 the first atomic-level explanation 1175:1208 the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19 1175:1313 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 7 17 theme increasing 1077:1086 arg1 dyad 1098:1101 an increasing catalytic dyad 1074:1101 an increasing catalytic dyad organization 1074:1114 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 1 18 theme Gaucher 95:101 arg1 disorder 135:142 an autosomal recessive disorder 112:142 an autosomal recessive disorder 112:142 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 18 theme Gaucher 95:101 arg1 disease 103:109 Gaucher disease 95:109 Gaucher disease 95:109 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 19 theme glucocerebrosidase 174:191 arg1 glycoprotein 242:253 a peripheral membrane-associated glycoprotein 209:253 a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 209:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 19 theme glucocerebrosidase 174:191 arg1 enzyme 201:206 glucocerebrosidase (GCase) enzyme 174:206 glucocerebrosidase (GCase) enzyme 174:206 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 3 20 theme GCase 508:512 arg1 activity 514:521 GCase activity 508:521 GCase activity 508:521 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 1 21 theme GCase 194:198 arg1 glycoprotein 242:253 a peripheral membrane-associated glycoprotein 209:253 a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 209:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 21 theme GCase 194:198 arg1 enzyme 201:206 glucocerebrosidase (GCase) enzyme 174:206 glucocerebrosidase (GCase) enzyme 174:206 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 0 22 theme proper 31:36 arg1 organization 53:64 a proper catalytic site organization 29:64 a proper catalytic site organization in human glucocerebrosidase 29:92 Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. 27023912 3 23 theme N-glycosylation 487:501 arg1 relevance 474:482 the relevance 470:482 the relevance of N-glycosylation over GCase activity 470:521 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 6 24 theme GCase 1038:1042 arg1 mutations 1044:1052 the most prevalent GCase mutations 1019:1052 the most prevalent GCase mutations 1019:1052 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 4 25 theme present 630:636 arg1 work 638:641 the present work 626:641 the present work 626:641 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 2 26 theme located 415:421 arg1 site 409:412 the first site 399:412 the first site 399:412 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 5 27 theme local 807:811 arg1 effects 842:848 local conformational stabilization effects 807:848 local conformational stabilization effects 807:848 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 4 28 theme triplicate 708:717 arg1 simulations 747:757 triplicate unbiased molecular dynamics simulations 708:757 triplicate unbiased molecular dynamics simulations 708:757 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 1 29 theme enzyme 201:206 arg1 deficiency 160:169 a deficiency 158:169 a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 158:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 0 30 theme site 48:51 arg1 organization 53:64 a proper catalytic site organization 29:64 a proper catalytic site organization in human glucocerebrosidase 29:92 Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. 27023912 5 31 theme enzyme 903:908 arg1 dyad 920:923 the enzyme catalytic dyad 899:923 the enzyme catalytic dyad 899:923 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 6 32 theme prevalent 1028:1036 arg1 mutations 1044:1052 the most prevalent GCase mutations 1019:1052 the most prevalent GCase mutations 1019:1052 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 0 33 from organization 53:64 arg1 glucocerebrosidase 75:92 human glucocerebrosidase 69:92 human glucocerebrosidase 69:92 Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. 27023912 4 34 theme unbiased 719:726 arg1 simulations 747:757 triplicate unbiased molecular dynamics simulations 708:757 triplicate unbiased molecular dynamics simulations 708:757 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 0 35 theme catalytic 38:46 arg1 organization 53:64 a proper catalytic site organization 29:64 a proper catalytic site organization in human glucocerebrosidase 29:92 Glycosylation is crucial for a proper catalytic site organization in human glucocerebrosidase. 27023912 6 36 theme Asn19-linked 945:956 arg1 glycan 958:963 the Asn19-linked glycan 941:963 the Asn19-linked glycan 941:963 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 2 37 from site 409:412 arg1 essential 319:327 essential 319:327 essential 319:327 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 1 38 theme autosomal 115:123 arg1 disorder 135:142 an autosomal recessive disorder 112:142 an autosomal recessive disorder 112:142 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 38 theme autosomal 115:123 arg1 disease 103:109 Gaucher disease 95:109 Gaucher disease 95:109 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 7 39 theme glycosylation 1142:1154 arg1 contents 1156:1163 glycosylation contents 1142:1163 glycosylation contents 1142:1163 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 6 40 link Asn19-linked 945:956 arg1 glycan 958:963 the Asn19-linked glycan 941:963 the Asn19-linked glycan 941:963 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 5 41 theme N-linked 777:784 arg1 chains 793:798 the N-linked glycan chains 773:798 the N-linked glycan chains 773:798 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 1 42 theme recessive 125:133 arg1 disorder 135:142 an autosomal recessive disorder 112:142 an autosomal recessive disorder 112:142 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 42 theme recessive 125:133 arg1 disease 103:109 Gaucher disease 95:109 Gaucher disease 95:109 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 6 43 theme glycan 958:963 arg1 case 933:936 the case 929:936 the case of the Asn19-linked glycan 929:963 In the case of the Asn19-linked glycan, it also occurred around region 438-444, where one of the most prevalent GCase mutations is found. 27023912 3 44 theme relevance 474:482 arg1 basis 461:465 the molecular basis 447:465 the molecular basis of the relevance of N-glycosylation over GCase activity 447:521 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 3 44 theme relevance 474:482 arg1 effects 531:537 the effects 527:537 the effects of glycosylation over its structure and dynamics 527:586 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 4 45 theme dynamics 738:745 arg1 simulations 747:757 triplicate unbiased molecular dynamics simulations 708:757 triplicate unbiased molecular dynamics simulations 708:757 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 7 46 theme GCase 1248:1252 arg1 activity 1254:1261 GCase activity 1248:1261 GCase activity 1248:1261 Markedly, an increasing catalytic dyad organization was related to increasing glycosylation contents, offering the first atomic-level explanation for the experimental observation that GCase activity is controlled by glycosylation, especially at Asn19. 27023912 3 47 theme molecular 451:459 arg1 basis 461:465 the molecular basis 447:465 the molecular basis of the relevance of N-glycosylation over GCase activity 447:521 However, both the molecular basis of the relevance of N-glycosylation over GCase activity and the effects of glycosylation over its structure and dynamics are still not fully understood. 27023912 4 48 theme glycosylation 680:692 arg1 content 694:700 glycosylation content 680:700 glycosylation content 680:700 Thus, the present work evaluated GCase enzyme in increasing glycosylation content using triplicate unbiased molecular dynamics simulations. 27023912 1 49 theme peripheral 211:220 arg1 glycoprotein 242:253 a peripheral membrane-associated glycoprotein 209:253 a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 209:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 49 theme peripheral 211:220 arg1 enzyme 201:206 glucocerebrosidase (GCase) enzyme 174:206 glucocerebrosidase (GCase) enzyme 174:206 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 5 50 theme conformational 813:826 arg1 effects 842:848 local conformational stabilization effects 807:848 local conformational stabilization effects 807:848 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 2 51 from Asn19 426:430 arg1 located 415:421 located 415:421 located 415:421 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 5 52 theme catalytic 910:918 arg1 dyad 920:923 the enzyme catalytic dyad 899:923 the enzyme catalytic dyad 899:923 Accordingly, the N-linked glycan chains caused local conformational stabilization effects over the protein, as well as in regions flanking the enzyme catalytic dyad. 27023912 2 53 from essential 319:327 arg1 site 409:412 the first site 399:412 the first site 399:412 Glycosylation is essential for the development of a catalytically active enzyme, specifically in the first site, located at Asn19. 27023912 1 54 theme membrane-associated 222:240 arg1 glycoprotein 242:253 a peripheral membrane-associated glycoprotein 209:253 a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes 209:299 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 27023912 1 54 theme membrane-associated 222:240 arg1 enzyme 201:206 glucocerebrosidase (GCase) enzyme 174:206 glucocerebrosidase (GCase) enzyme 174:206 Gaucher disease, an autosomal recessive disorder, is caused by a deficiency of glucocerebrosidase (GCase) enzyme, a peripheral membrane-associated glycoprotein that hydrolyses glucosylceramide in lysosomes. 26563299 7 0 gly N-glycosylation 1091:1105 arg2 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 0 gly N-glycosylation 1091:1105 arg2 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 0 gly N-glycosylation 1091:1105 arg2 unoccupied 1176:1185 unoccupied 1176:1185 unoccupied 1176:1185 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 0 gly N-glycosylation 1091:1105 arg2 two 1071:1073 two 1071:1073 two 1071:1073 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 0 gly N-glycosylation 1091:1105 arg2 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 14 1 theme clinical 2129:2136 arg1 efficacy 2138:2145 clinical efficacy 2129:2145 clinical efficacy 2129:2145 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 6 2 theme IL-15 881:885 arg1 Asn79 887:891 IL-15 Asn79 881:891 IL-15 Asn79 881:891 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 5 3 link derived 786:792 arg1 glycans 794:800 derived glycans 786:800 derived glycans 786:800 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 6 4 theme N-glycans 969:977 arg1 N-glycans 969:977 biosynthetically-related N-glycans 944:977 biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 944:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 6 4 theme N-glycans 969:977 arg1 repertoire 930:939 the same repertoire 921:939 the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 921:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 4 5 theme relevant 676:683 arg1 preparations 697:708 two clinically relevant large-scale preparations 661:708 two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15 661:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 4 6 gly O-glycosylation 642:656 arg1 preparations 697:708 two clinically relevant large-scale preparations 661:708 two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15 661:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 2 7 theme IL-15 261:265 arg1 complex 276:282 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 1 8 theme stable 167:172 arg1 complex 184:190 a stable molecular complex 165:190 a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα) 165:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 13 9 gly O-glycosylation 1929:1943 arg1 preparations 1960:1971 large-scale preparations 1948:1971 large-scale preparations of the therapeutic candidate hetIL-15 1948:2009 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 13 9 gly O-glycosylation 1929:1943 arg1 conclusion 1863:1872 conclusion 1863:1872 conclusion 1863:1872 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 9 10 gly sialylated 1414:1423 arg1 core 1425:1428 core 1 1425:1430 core 1 1425:1430 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 14 11 theme hetIL-15 2150:2157 arg1 efficacy 2138:2145 clinical efficacy 2129:2145 clinical efficacy 2129:2145 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 14 11 theme hetIL-15 2150:2157 arg1 production 2114:2123 consistent large-scale production 2091:2123 consistent large-scale production 2091:2123 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 12 12 theme batches 1784:1790 arg1 glycosylation 1741:1753 Highly reproducible glycosylation 1721:1753 Highly reproducible glycosylation of IL-15 1721:1762 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 12 12 theme batches 1784:1790 arg1 sIL-15Rα 1768:1775 sIL-15Rα 1768:1775 sIL-15Rα of two batches of hetIL-15 1768:1802 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 4 13 link HEK293-derived 713:726 arg1 hetIL-15 746:753 HEK293-derived recombinant human hetIL-15 713:753 HEK293-derived recombinant human hetIL-15 713:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 13 14 theme candidate 1992:2000 arg1 hetIL-15 2002:2009 the therapeutic candidate hetIL-15 1976:2009 the therapeutic candidate hetIL-15 1976:2009 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Thr81 1477:1481 Thr81 1477:1481 Thr81 1477:1481 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Ser158 1499:1504 Ser158 1499:1504 Ser158 1499:1504 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Ser160 1511:1516 Ser160 1511:1516 Ser160 1511:1516 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Thr86 1484:1488 Thr86 1484:1488 Thr86 1484:1488 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Thr156 1491:1496 Thr156 1491:1496 Thr156 1491:1496 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 15 from hexasaccharides 1452:1466 arg1 Thr2 1471:1474 Thr2 1471:1474 Thr2 1471:1474 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 12 16 theme hetIL-15 1795:1802 arg1 batches 1784:1790 two batches 1780:1790 two batches of hetIL-15 1780:1802 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 12 17 gly glycosylation 1741:1753 arg1 batches 1784:1790 two batches 1780:1790 two batches of hetIL-15 1780:1802 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 12 17 gly glycosylation 1741:1753 arg1 IL-15 1758:1762 IL-15 1758:1762 IL-15 1758:1762 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 7 18 theme located 1132:1138 arg1 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 18 theme located 1132:1138 arg1 unoccupied 1176:1185 unoccupied 1176:1185 unoccupied 1176:1185 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 18 theme located 1132:1138 arg1 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 18 theme located 1132:1138 arg1 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 1 19 theme IL-15 209:213 arg1 sIL-15Rα 231:238 sIL-15Rα 231:238 sIL-15Rα 231:238 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 1 19 theme IL-15 209:213 arg1 alpha 224:228 the soluble IL-15 receptor alpha 197:228 the soluble IL-15 receptor alpha (sIL-15Rα) 197:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 11 20 dep Non-human 1585:1593 arg1 immunogenic 1608:1618 immunogenic 1608:1618 immunogenic 1608:1618 Non-human, potentially immunogenic glycoepitopes (e.g. N-glycolylneuraminic acid and α-galactosylation) were not displayed by hetIL-15. 26563299 6 21 contain carried 913:919 arg2 repertoire 930:939 the same repertoire 921:939 the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 921:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 6 21 contain carried 913:919 arg1 Asn79 887:891 IL-15 Asn79 881:891 IL-15 Asn79 881:891 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 6 21 contain carried 913:919 arg1 Asn107 906:911 sIL-15Rα Asn107 897:911 sIL-15Rα Asn107 897:911 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 6 21 contain carried 913:919 arg2 N-glycans 969:977 biosynthetically-related N-glycans 944:977 biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 944:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 13 22 theme preparations 1960:1971 arg1 O-glycosylation 1929:1943 O-glycosylation 1929:1943 O-glycosylation 1929:1943 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 13 22 theme preparations 1960:1971 arg1 N- 1922:1923 N- 1922:1923 N- 1922:1923 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 14 23 theme large-scale 2102:2112 arg1 production 2114:2123 consistent large-scale production 2091:2123 consistent large-scale production 2091:2123 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 6 24 theme complex-type 1042:1053 arg1 structures 1055:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 7 25 theme IL-2 1147:1150 arg1 interface 1161:1169 the IL-2 receptor interface 1143:1169 the IL-2 receptor interface 1143:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 12 26 theme reproducible 1728:1739 arg1 glycosylation 1741:1753 Highly reproducible glycosylation 1721:1753 Highly reproducible glycosylation of IL-15 1721:1762 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 4 27 theme HEK293-derived 713:726 arg1 hetIL-15 746:753 HEK293-derived recombinant human hetIL-15 713:753 HEK293-derived recombinant human hetIL-15 713:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 0 28 theme reproducible 69:80 arg1 N- 82:83 extensive and reproducible N- 55:83 extensive and reproducible N- 55:83 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 4 29 theme human 740:744 arg1 hetIL-15 746:753 HEK293-derived recombinant human hetIL-15 713:753 HEK293-derived recombinant human hetIL-15 713:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 7 30 theme IL-15 1085:1089 arg1 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 30 theme IL-15 1085:1089 arg1 unoccupied 1176:1185 unoccupied 1176:1185 unoccupied 1176:1185 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 30 theme IL-15 1085:1089 arg1 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 30 theme IL-15 1085:1089 arg1 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 11 31 dep acid 1661:1664 arg1 e.g. 1635:1638 e.g. 1635:1638 e.g. 1635:1638 Non-human, potentially immunogenic glycoepitopes (e.g. N-glycolylneuraminic acid and α-galactosylation) were not displayed by hetIL-15. 26563299 3 32 gly glycosylation 531:543 arg1 hetIL-15 548:555 hetIL-15 548:555 hetIL-15 548:555 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 6 33 gly α1-6-core-fucosylated 995:1015 arg1 structures 1055:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 2 34 theme sIL-15Rα 267:274 arg1 complex 276:282 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 2 35 theme lymphocytes 379:389 arg1 growth 340:345 growth 340:345 growth 340:345 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 2 35 theme lymphocytes 379:389 arg1 activation 365:374 activation 365:374 activation 365:374 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 2 35 theme lymphocytes 379:389 arg1 mobilization 348:359 mobilization 348:359 mobilization 348:359 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 5 36 theme derived 786:792 arg1 glycans 794:800 derived glycans 786:800 derived glycans 786:800 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 11 37 theme Non-human 1585:1593 arg1 glycoepitopes 1620:1632 Non-human, potentially immunogenic glycoepitopes 1585:1632 Non-human, potentially immunogenic glycoepitopes (e.g. N-glycolylneuraminic acid and α-galactosylation) 1585:1687 Non-human, potentially immunogenic glycoepitopes (e.g. N-glycolylneuraminic acid and α-galactosylation) were not displayed by hetIL-15. 26563299 0 38 theme Recombinant 0:10 arg1 complex 38:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 14 39 theme features 2053:2060 arg1 mapping 2026:2032 Site-specific mapping 2012:2032 Site-specific mapping of these molecular features 2012:2060 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 2 40 theme clinical 421:428 arg1 trials 430:435 clinical trials 421:435 clinical trials 421:435 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 0 41 theme heterodimeric 18:30 arg1 complex 38:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 3 42 theme Favorable 438:446 arg1 properties 464:473 Favorable pharmacokinetic properties 438:473 Favorable pharmacokinetic properties 438:473 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 8 43 theme intact 1205:1210 arg1 IL-15 1212:1216 intact IL-15 1205:1216 intact IL-15 1205:1216 Mass analysis of intact IL-15 confirmed its N-glycosylation and suggested that Asn79-glycosylation partially prevents Asn77-deamidation. 26563299 0 44 link O-linked 89:96 arg1 glycosylation 98:110 O-linked glycosylation 89:110 O-linked glycosylation 89:110 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 5 45 theme multiple 851:858 arg1 strategies 869:878 multiple LC-MS/MS strategies 851:878 multiple LC-MS/MS strategies 851:878 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 1 46 with complex 184:190 arg1 sIL-15Rα 231:238 sIL-15Rα 231:238 sIL-15Rα 231:238 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 1 46 with complex 184:190 arg1 alpha 224:228 the soluble IL-15 receptor alpha 197:228 the soluble IL-15 receptor alpha (sIL-15Rα) 197:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 10 47 dep α2-3- 1546:1550 arg1 sialylation 1572:1582 sialylation 1572:1582 sialylation 1572:1582 The sialoglycans displayed α2-3- and α2-6-NeuAc-type sialylation. 26563299 1 48 theme receptor 215:222 arg1 sIL-15Rα 231:238 sIL-15Rα 231:238 sIL-15Rα 231:238 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 1 48 theme receptor 215:222 arg1 alpha 224:228 the soluble IL-15 receptor alpha 197:228 the soluble IL-15 receptor alpha (sIL-15Rα) 197:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 3 49 theme hetIL-15 548:555 arg1 formation 513:521 the heterodimeric formation 495:521 the heterodimeric formation 495:521 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 3 49 theme hetIL-15 548:555 arg1 glycosylation 531:543 the glycosylation 527:543 the glycosylation of hetIL-15 527:555 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 6 50 theme β-GlcNAc-terminating 1021:1040 arg1 structures 1055:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 7 51 from interface 1161:1169 arg1 located 1132:1138 located 1132:1138 located 1132:1138 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 14 52 dep production 2114:2123 arg1 the 2087:2089 the 2087:2089 the 2087:2089 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 6 53 theme α1-6-core-fucosylated 995:1015 arg1 structures 1055:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 988:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 13 54 theme hetIL-15 2002:2009 arg1 preparations 1960:1971 large-scale preparations 1948:1971 large-scale preparations of the therapeutic candidate hetIL-15 1948:2009 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 4 55 theme preparations 697:708 arg1 O-glycosylation 642:656 O-glycosylation 642:656 O-glycosylation 642:656 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 4 55 theme preparations 697:708 arg1 N- 635:636 the site-specific N- 617:636 the site-specific N- 617:636 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 0 56 theme extensive 55:63 arg1 N- 82:83 extensive and reproducible N- 55:83 extensive and reproducible N- 55:83 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 6 57 theme biosynthetically-related 944:967 arg1 N-glycans 969:977 biosynthetically-related N-glycans 944:977 biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 944:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 9 58 gly O-glycosylated 1384:1397 arg1 sIL-15Rα 1363:1370 sIL-15Rα 1363:1370 sIL-15Rα 1363:1370 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 1 59 theme molecular 174:182 arg1 complex 184:190 a stable molecular complex 165:190 a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα) 165:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 4 60 theme large-scale 685:695 arg1 preparations 697:708 two clinically relevant large-scale preparations 661:708 two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15 661:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 13 61 theme therapeutic 1980:1990 arg1 hetIL-15 2002:2009 the therapeutic candidate hetIL-15 1976:2009 the therapeutic candidate hetIL-15 1976:2009 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 5 62 theme Intact 756:761 arg1 IL-15 763:767 IL-15 763:767 IL-15 763:767 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 1 63 theme soluble 201:207 arg1 sIL-15Rα 231:238 sIL-15Rα 231:238 sIL-15Rα 231:238 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 1 63 theme soluble 201:207 arg1 alpha 224:228 the soluble IL-15 receptor alpha 197:228 the soluble IL-15 receptor alpha (sIL-15Rα) 197:239 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 13 64 theme large-scale 1948:1958 arg1 preparations 1960:1971 large-scale preparations 1948:1971 large-scale preparations of the therapeutic candidate hetIL-15 1948:2009 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 7 65 theme receptor 1152:1159 arg1 interface 1161:1169 the IL-2 receptor interface 1143:1169 the IL-2 receptor interface 1143:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 9 66 theme sialylated 1414:1423 arg1 core 1425:1428 core 1 1425:1430 core 1 1425:1430 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 4 67 theme hetIL-15 746:753 arg1 preparations 697:708 two clinically relevant large-scale preparations 661:708 two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15 661:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 2 68 theme heterodimeric 247:259 arg1 complex 276:282 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) 242:293 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 14 69 theme consistent 2091:2100 arg1 production 2114:2123 consistent large-scale production 2091:2123 consistent large-scale production 2091:2123 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 11 70 theme N-glycolylneuraminic 1640:1659 arg1 acid 1661:1664 N-glycolylneuraminic acid 1640:1664 N-glycolylneuraminic acid 1640:1664 Non-human, potentially immunogenic glycoepitopes (e.g. N-glycolylneuraminic acid and α-galactosylation) were not displayed by hetIL-15. 26563299 9 71 theme 2-type 1436:1441 arg1 mono- 1443:1447 2-type mono- 1436:1447 2-type mono- 1436:1447 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 12 72 theme IL-15 1758:1762 arg1 glycosylation 1741:1753 Highly reproducible glycosylation 1721:1753 Highly reproducible glycosylation of IL-15 1721:1762 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 12 72 theme IL-15 1758:1762 arg1 sIL-15Rα 1768:1775 sIL-15Rα 1768:1775 sIL-15Rα of two batches of hetIL-15 1768:1802 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 0 73 theme O-linked 89:96 arg1 glycosylation 98:110 O-linked glycosylation 89:110 O-linked glycosylation 89:110 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 13 74 theme reproducible 1909:1920 arg1 N- 1922:1923 N- 1922:1923 N- 1922:1923 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 4 75 theme recombinant 728:738 arg1 hetIL-15 746:753 HEK293-derived recombinant human hetIL-15 713:753 HEK293-derived recombinant human hetIL-15 713:753 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 7 76 theme N-glycosylation 1091:1105 arg1 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 76 theme N-glycosylation 1091:1105 arg1 unoccupied 1176:1185 unoccupied 1176:1185 unoccupied 1176:1185 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 76 theme N-glycosylation 1091:1105 arg1 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 76 theme N-glycosylation 1091:1105 arg1 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 77 theme potential 1075:1083 arg1 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 77 theme potential 1075:1083 arg1 unoccupied 1176:1185 unoccupied 1176:1185 unoccupied 1176:1185 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 77 theme potential 1075:1083 arg1 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 77 theme potential 1075:1083 arg1 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 5 78 gly glycopeptides 806:818 arg2 glycopeptides 806:818 glycopeptides 806:818 glycopeptides 806:818 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 0 79 theme human 12:16 arg1 complex 38:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 8 80 theme Mass 1188:1191 arg1 analysis 1193:1200 Mass analysis 1188:1200 Mass analysis of intact IL-15 1188:1216 Mass analysis of intact IL-15 confirmed its N-glycosylation and suggested that Asn79-glycosylation partially prevents Asn77-deamidation. 26563299 13 81 theme heterogeneous 1891:1903 arg1 N- 1922:1923 N- 1922:1923 N- 1922:1923 In conclusion, we document the heterogeneous and reproducible N- and O-glycosylation of large-scale preparations of the therapeutic candidate hetIL-15. 26563299 7 82 dep sites 1107:1111 arg1 Asn71 1114:1118 Asn71 1114:1118 Asn71 1114:1118 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 82 dep sites 1107:1111 arg1 sites 1107:1111 The two potential IL-15 N-glycosylation sites 1067:1111 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface 1067:1169 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 7 82 dep sites 1107:1111 arg1 Asn112 1124:1129 Asn112 1124:1129 Asn112 1124:1129 The two potential IL-15 N-glycosylation sites (Asn71 and Asn112) located at the IL-2 receptor interface were unoccupied. 26563299 0 83 theme IL-15 32:36 arg1 complex 38:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex 0:44 Recombinant human heterodimeric IL-15 complex displays extensive and reproducible N- and O-linked glycosylation. 26563299 14 84 theme molecular 2043:2051 arg1 features 2053:2060 these molecular features 2037:2060 these molecular features 2037:2060 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 1 85 theme Human 113:117 arg1 IL-15 135:139 IL-15 135:139 IL-15 135:139 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 1 85 theme Human 113:117 arg1 interleukin 119:129 Human interleukin 15 113:132 Human interleukin 15 (IL-15) 113:140 Human interleukin 15 (IL-15) circulates in blood as a stable molecular complex with the soluble IL-15 receptor alpha (sIL-15Rα). 26563299 12 86 theme consistent 1817:1826 arg1 manufacturing 1828:1840 consistent manufacturing 1817:1840 consistent manufacturing 1817:1840 Highly reproducible glycosylation of IL-15 and sIL-15Rα of two batches of hetIL-15 demonstrated consistent manufacturing and purification. 26563299 9 87 with O-glycosylated 1384:1397 arg1 mono- 1443:1447 2-type mono- 1436:1447 2-type mono- 1436:1447 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 87 with O-glycosylated 1384:1397 arg1 core 1425:1428 core 1 1425:1430 core 1 1425:1430 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 8 88 theme IL-15 1212:1216 arg1 analysis 1193:1200 Mass analysis 1188:1200 Mass analysis of intact IL-15 1188:1216 Mass analysis of intact IL-15 confirmed its N-glycosylation and suggested that Asn79-glycosylation partially prevents Asn77-deamidation. 26563299 6 89 theme same 925:928 arg1 N-glycans 969:977 biosynthetically-related N-glycans 944:977 biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 944:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 6 89 theme same 925:928 arg1 repertoire 930:939 the same repertoire 921:939 the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures 921:1064 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 3 90 theme pharmacokinetic 448:462 arg1 properties 464:473 Favorable pharmacokinetic properties 438:473 Favorable pharmacokinetic properties 438:473 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 14 91 theme Site-specific 2012:2024 arg1 mapping 2026:2032 Site-specific mapping 2012:2032 Site-specific mapping of these molecular features 2012:2060 Site-specific mapping of these molecular features is important to evaluate the consistent large-scale production and clinical efficacy of hetIL-15. 26563299 3 92 theme heterodimeric 499:511 arg1 formation 513:521 the heterodimeric formation 495:521 the heterodimeric formation 495:521 Favorable pharmacokinetic properties are associated with the heterodimeric formation and the glycosylation of hetIL-15, which, however, remains largely uncharacterized. 26563299 6 93 theme sIL-15Rα 897:904 arg1 Asn107 906:911 sIL-15Rα Asn107 897:911 sIL-15Rα Asn107 897:911 IL-15 Asn79 and sIL-15Rα Asn107 carried the same repertoire of biosynthetically-related N-glycans covering mostly α1-6-core-fucosylated and β-GlcNAc-terminating complex-type structures. 26563299 2 94 theme therapeutic 301:311 arg1 potential 313:321 therapeutic potential 301:321 therapeutic potential 301:321 This heterodimeric IL-15:sIL-15Rα complex (hetIL-15) shows therapeutic potential by promoting the growth, mobilization and activation of lymphocytes and is currently evaluated in clinical trials. 26563299 4 95 theme site-specific 621:633 arg1 N- 635:636 the site-specific N- 617:636 the site-specific N- 617:636 We report the site-specific N- and O-glycosylation of two clinically relevant large-scale preparations of HEK293-derived recombinant human hetIL-15. 26563299 5 96 theme LC-MS/MS 860:867 arg1 strategies 869:878 multiple LC-MS/MS strategies 851:878 multiple LC-MS/MS strategies 851:878 Intact IL-15 and sIL-15Rα and derived glycans and glycopeptides were separately profiled using multiple LC-MS/MS strategies. 26563299 9 97 contain contained 1331:1339 arg2 O-glycans 1344:1352 no O-glycans 1341:1352 no O-glycans 1341:1352 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 26563299 9 97 contain contained 1331:1339 arg1 IL-15 1325:1329 IL-15 1325:1329 IL-15 1325:1329 IL-15 contained no O-glycans, whereas sIL-15Rα was heavily O-glycosylated with partially sialylated core 1 and 2-type mono- to hexasaccharides on Thr2, Thr81, Thr86, Thr156, Ser158, and Ser160. 25092234 6 0 theme chain 975:979 arg1 occupancy 952:960 full-site occupancy 942:960 full-site occupancy of the light chain 942:979 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 10 1 theme structural 1368:1377 arg1 information 1379:1389 valuable structural information 1359:1389 valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function 1359:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 3 2 theme glycosylation 462:474 arg1 analysis 476:483 the first in-depth glycosylation analysis 443:483 the first in-depth glycosylation analysis of hFXI based on advanced MS approaches 443:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 10 3 theme potential 1416:1424 arg1 roles 1426:1430 the potential roles 1412:1430 the potential roles of N-glycosylation on hFXI function 1412:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 7 4 theme hFXI 994:997 arg1 glycopeptides 999:1011 hFXI glycopeptides 994:1011 hFXI glycopeptides 994:1011 Analysis of hFXI glycopeptides by LC-MS/MS enabled site-specific glycan profiling and occupancy. 25092234 3 5 theme advanced 502:509 arg1 approaches 514:523 advanced MS approaches 502:523 advanced MS approaches 502:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 1 6 theme homodimer 161:169 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 6 theme homodimer 161:169 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 8 7 used occupied 1160:1167 arg2 N108 1144:1147 N108 1144:1147 N108 1144:1147 It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. 25092234 8 7 used occupied 1160:1167 arg2 N72 1136:1138 N72 1136:1138 N72 1136:1138 It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. 25092234 4 8 dep S2 743:744 arg1 % 750:750 66% 748:750 66% 748:750 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 6 9 theme intact 851:856 arg1 hFXI 858:861 intact hFXI 851:861 intact hFXI 851:861 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 9 10 theme noncanonical 1283:1294 arg1 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 4 11 theme N-glycans 603:611 arg1 MS 557:558 MS 557:558 MS 557:558 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 11 theme N-glycans 603:611 arg1 quantification 581:594 quantification 581:594 quantification of the N-glycans 581:611 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 11 theme N-glycans 603:611 arg1 LC 550:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 12 dep LC 550:551 arg1 characterization 560:575 characterization 560:575 characterization 560:575 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 5 13 theme S3 788:789 arg1 F 791:791 A3 S3 F 785:791 A3 S3 F 785:791 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 4 14 theme A2 691:692 arg1 S1 694:695 A2 S1 691:695 A2 S1 691:695 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 14 theme A2 691:692 arg1 % 701:701 20% 699:701 20% 699:701 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 15 gly mono-α2,6-sialylated 669:688 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 15 gly mono-α2,6-sialylated 669:688 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 15 gly mono-α2,6-sialylated 669:688 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 16 theme complex 649:655 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 16 theme complex 649:655 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 16 theme complex 649:655 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 5 17 theme A3 801:802 arg1 S3 804:805 A3 S3 801:805 A3 S3 801:805 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 6 18 theme two 891:893 arg1 occupation 877:886 full occupation 872:886 full occupation of two of the three heavy-chain glycosites 872:929 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 4 19 theme mono-α2,6-sialylated 669:688 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 19 theme mono-α2,6-sialylated 669:688 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 19 theme mono-α2,6-sialylated 669:688 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 1 20 theme 160-kDa 135:141 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 20 theme 160-kDa 135:141 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 6 21 dep occupation 877:886 arg1 occupancy 952:960 full-site occupancy 942:960 full-site occupancy of the light chain 942:979 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 0 22 theme NXC 92:94 arg1 glycosite 96:104 a noncanonical NXC glycosite 77:104 a noncanonical NXC glycosite 77:104 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 2 23 theme hemophilia 276:285 arg1 hFXI 290:293 hemophilia C. hFXI 276:293 hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain 276:421 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 4 24 theme A2 740:741 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 24 theme A2 740:741 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 0 25 theme factor 48:53 arg1 XI 55:56 human factor XI 42:56 human factor XI 42:56 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 1 26 link disulphide-linked 143:159 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 26 link disulphide-linked 143:159 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 0 27 theme Site-specific 0:12 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 0:105 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 0 28 gly glycosite 96:104 arg2 glycosite 96:104 a noncanonical NXC glycosite 77:104 a noncanonical NXC glycosite 77:104 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 6 29 theme full-site 942:950 arg1 occupancy 952:960 full-site occupancy 942:960 full-site occupancy of the light chain 942:979 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 5 30 theme triantennary 760:771 arg1 structures 773:782 Minor triantennary structures 754:782 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) 754:812 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 1 31 theme Human 107:111 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 31 theme Human 107:111 arg1 hFXI 124:127 hFXI 124:127 hFXI 124:127 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 31 theme Human 107:111 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 7 32 theme glycan 1047:1052 arg1 profiling 1054:1062 site-specific glycan profiling 1033:1062 site-specific glycan profiling 1033:1062 Analysis of hFXI glycopeptides by LC-MS/MS enabled site-specific glycan profiling and occupancy. 25092234 0 33 theme human 42:46 arg1 XI 55:56 human factor XI 42:56 human factor XI 42:56 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 0 34 gly N-glycosylation 14:28 arg1 XI 55:56 human factor XI 42:56 human factor XI 42:56 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 6 35 theme heavy-chain 908:918 arg1 glycosites 920:929 the three heavy-chain glycosites 898:929 the three heavy-chain glycosites 898:929 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 10 36 theme valuable 1359:1366 arg1 information 1379:1389 valuable structural information 1359:1389 valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function 1359:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 5 37 dep F 791:791 arg1 % 811:811 ∼2% 809:811 ∼2% 809:811 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 5 37 dep F 791:791 arg1 % 798:798 ∼1.5% 794:798 ∼1.5% 794:798 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 0 38 theme XI 55:56 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 0:105 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 9 39 gly glycosite 1266:1274 arg1 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 9 39 gly glycosite 1266:1274 arg2 glycosite 1266:1274 a new glycosite 1260:1274 a new glycosite of the noncanonical format NXC 1260:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 9 39 gly glycosite 1266:1274 arg2 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 10 40 from roles 1426:1430 arg1 function 1459:1466 hFXI function 1454:1466 hFXI function 1454:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 0 41 dep analysis 30:37 arg1 Identification 59:72 Identification 59:72 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 0:105 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 5 42 dep structures 773:782 arg1 F 791:791 A3 S3 F 785:791 A3 S3 F 785:791 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 9 43 theme format 1296:1301 arg1 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 7 44 theme glycopeptides 999:1011 arg1 Analysis 982:989 Analysis 982:989 Analysis of hFXI glycopeptides by LC-MS/MS 982:1023 Analysis of hFXI glycopeptides by LC-MS/MS enabled site-specific glycan profiling and occupancy. 25092234 3 45 theme in-depth 453:460 arg1 analysis 476:483 the first in-depth glycosylation analysis 443:483 the first in-depth glycosylation analysis of hFXI based on advanced MS approaches 443:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 9 46 theme new 1262:1264 arg1 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 9 46 theme new 1262:1264 arg1 glycosite 1266:1274 a new glycosite 1260:1274 a new glycosite of the noncanonical format NXC 1260:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 6 47 theme light 969:973 arg1 chain 975:979 the light chain 965:979 the light chain 965:979 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 4 48 theme interaction 538:548 arg1 LC 550:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 2 49 theme C. 287:288 arg1 hFXI 290:293 hemophilia C. hFXI 276:293 hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain 276:421 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 3 50 theme hFXI 488:491 arg1 analysis 476:483 the first in-depth glycosylation analysis 443:483 the first in-depth glycosylation analysis of hFXI based on advanced MS approaches 443:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 1 51 theme disulphide-linked 143:159 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 51 theme disulphide-linked 143:159 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 10 52 theme hFXI 1454:1457 arg1 function 1459:1466 hFXI function 1454:1466 hFXI function 1454:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 2 53 theme N-glycosylation 306:320 arg1 N335 364:367 N335 364:367 N335 364:367 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 53 theme N-glycosylation 306:320 arg1 N108 357:360 N108 357:360 N108 357:360 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 53 theme N-glycosylation 306:320 arg1 sites 332:336 five N-glycosylation consensus sites 301:336 five N-glycosylation consensus sites per monomer 301:348 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 53 theme N-glycosylation 306:320 arg1 N473 399:402 N473 399:402 N473 399:402 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 9 54 theme NXC 1303:1305 arg1 NXC 1303:1305 the noncanonical format NXC 1279:1305 the noncanonical format NXC 1279:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 9 54 theme NXC 1303:1305 arg1 glycosite 1266:1274 a new glycosite 1260:1274 a new glycosite of the noncanonical format NXC 1260:1305 We also identified a new glycosite of the noncanonical format NXC at N145 , occupied at around 5%. 25092234 6 55 theme hFXI 858:861 arg1 analyses 839:846 MS analyses 836:846 MS analyses of intact hFXI 836:861 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 4 56 theme major 633:637 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 56 theme major 633:637 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 10 57 theme N-glycosylation 1435:1449 arg1 roles 1426:1430 the potential roles 1412:1430 the potential roles of N-glycosylation on hFXI function 1412:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 8 58 gly glycosylated 1114:1125 arg1 N335 1101:1104 N335 1101:1104 N335 1101:1104 It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. 25092234 3 59 theme MS 511:512 arg1 approaches 514:523 advanced MS approaches 502:523 advanced MS approaches 502:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 7 60 gly glycopeptides 999:1011 arg2 glycopeptides 999:1011 hFXI glycopeptides 994:1011 hFXI glycopeptides 994:1011 Analysis of hFXI glycopeptides by LC-MS/MS enabled site-specific glycan profiling and occupancy. 25092234 5 61 theme A3 785:786 arg1 F 791:791 A3 S3 F 785:791 A3 S3 F 785:791 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 6 62 theme MS 836:837 arg1 analyses 839:846 MS analyses 836:846 MS analyses of intact hFXI 836:861 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 1 63 theme factor 113:118 arg1 zymogen 171:177 a 160-kDa disulphide-linked homodimer zymogen 133:177 a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade 133:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 63 theme factor 113:118 arg1 hFXI 124:127 hFXI 124:127 hFXI 124:127 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 1 63 theme factor 113:118 arg1 XI 120:121 Human factor XI 107:121 Human factor XI (hFXI) 107:128 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 2 64 dep hFXI 290:293 arg1 bears 295:299 bears 295:299 bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain 295:421 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 8 65 used occupied 1197:1204 arg2 N473 1187:1190 N473 1187:1190 N473 1187:1190 It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. 25092234 8 65 used occupied 1197:1204 arg2 N432 1178:1181 N432 1178:1181 N432 1178:1181 It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. 25092234 4 66 gly bis-α2,6-sialylated 708:726 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 66 gly bis-α2,6-sialylated 708:726 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 66 gly bis-α2,6-sialylated 708:726 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 2 67 theme bleeding 242:249 arg1 diathesis 251:259 bleeding diathesis 242:259 bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain 242:421 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 0 68 theme noncanonical 79:90 arg1 glycosite 96:104 a noncanonical NXC glycosite 77:104 a noncanonical NXC glycosite 77:104 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 4 69 theme biantennary 657:667 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 69 theme biantennary 657:667 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 69 theme biantennary 657:667 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 0 70 theme glycosite 96:104 arg1 Identification 59:72 Identification 59:72 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 0:105 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 4 71 dep mono-α2,6-sialylated 669:688 arg1 S1 694:695 A2 S1 691:695 A2 S1 691:695 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 71 dep mono-α2,6-sialylated 669:688 arg1 % 701:701 20% 699:701 20% 699:701 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 6 72 theme full 872:875 arg1 occupation 877:886 full occupation 872:886 full occupation of two of the three heavy-chain glycosites 872:929 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 4 73 theme bis-α2,6-sialylated 708:726 arg1 structures 728:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures 649:737 complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%) 649:751 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 73 theme bis-α2,6-sialylated 708:726 arg1 S2 743:744 A2 S2 740:744 A2 S2 740:744 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 4 73 theme bis-α2,6-sialylated 708:726 arg1 forms 639:643 the two major forms 625:643 the two major forms 625:643 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 10 74 theme structural 1489:1498 arg1 data 1346:1349 These data 1340:1349 These data 1340:1349 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 10 74 theme structural 1489:1498 arg1 reference 1500:1508 a structural reference 1487:1508 a structural reference 1487:1508 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 0 75 theme N-glycosylation 14:28 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 0:105 Site-specific N-glycosylation analysis of human factor XI: Identification of a noncanonical NXC glycosite. 25092234 5 76 theme Minor 754:758 arg1 structures 773:782 Minor triantennary structures 754:782 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) 754:812 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 4 77 theme Hydrophilic 526:536 arg1 LC 550:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC 526:551 Hydrophilic interaction LC and MS characterization and quantification of the N-glycans showed that the two major forms are complex biantennary mono-α2,6-sialylated (A2 S1 , 20%) and bis-α2,6-sialylated structures (A2 S2 , 66%). 25092234 3 78 gly glycosylation 462:474 arg1 hFXI 488:491 hFXI 488:491 hFXI 488:491 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 7 79 theme site-specific 1033:1045 arg1 profiling 1054:1062 site-specific glycan profiling 1033:1062 site-specific glycan profiling 1033:1062 Analysis of hFXI glycopeptides by LC-MS/MS enabled site-specific glycan profiling and occupancy. 25092234 1 80 theme coagulation 195:205 arg1 cascade 207:213 the coagulation cascade 191:213 the coagulation cascade 191:213 Human factor XI (hFXI) is a 160-kDa disulphide-linked homodimer zymogen involved in the coagulation cascade. 25092234 6 81 theme glycosites 920:929 arg1 glycosites 920:929 the three heavy-chain glycosites 898:929 the three heavy-chain glycosites 898:929 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 6 81 theme glycosites 920:929 arg1 two 891:893 two 891:893 two 891:893 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 6 82 gly glycosites 920:929 arg2 glycosites 920:929 the three heavy-chain glycosites 898:929 the three heavy-chain glycosites 898:929 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 5 83 dep % 798:798 arg1 S3 804:805 A3 S3 801:805 A3 S3 801:805 Minor triantennary structures (A3 S3 F, ∼1.5%; A3 S3 , ∼2%) were also identified. 25092234 2 84 theme heavy 376:380 arg1 chain 382:386 the heavy chain 372:386 the heavy chain 372:386 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 10 85 theme useful 1391:1396 arg1 information 1379:1389 valuable structural information 1359:1389 valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function 1359:1466 These data provide valuable structural information useful to understand the potential roles of N-glycosylation on hFXI function and could serve as a structural reference. 25092234 2 86 theme light 411:415 arg1 chain 417:421 the light chain 407:421 the light chain 407:421 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 3 87 theme first 447:451 arg1 analysis 476:483 the first in-depth glycosylation analysis 443:483 the first in-depth glycosylation analysis of hFXI based on advanced MS approaches 443:523 This study reports the first in-depth glycosylation analysis of hFXI based on advanced MS approaches. 25092234 6 88 gly occupancy 952:960 arg2 chain 975:979 the light chain 965:979 the light chain 965:979 MS analyses of intact hFXI revealed full occupation of two of the three heavy-chain glycosites and almost full-site occupancy of the light chain. 25092234 2 89 theme consensus 322:330 arg1 N335 364:367 N335 364:367 N335 364:367 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 89 theme consensus 322:330 arg1 N108 357:360 N108 357:360 N108 357:360 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 89 theme consensus 322:330 arg1 sites 332:336 five N-glycosylation consensus sites 301:336 five N-glycosylation consensus sites per monomer 301:348 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 89 theme consensus 322:330 arg1 N473 399:402 N473 399:402 N473 399:402 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 90 gly N-glycosylation 306:320 arg2 N108 357:360 N108 357:360 N108 357:360 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 90 gly N-glycosylation 306:320 arg2 N335 364:367 N335 364:367 N335 364:367 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 90 gly N-glycosylation 306:320 arg2 five 301:304 five 301:304 five 301:304 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 90 gly N-glycosylation 306:320 arg2 N473 399:402 N473 399:402 N473 399:402 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 25092234 2 90 gly N-glycosylation 306:320 arg2 sites 332:336 five N-glycosylation consensus sites 301:336 five N-glycosylation consensus sites per monomer 301:348 Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N-glycosylation consensus sites per monomer, N72 , N108 , N335 on the heavy chain and N432 , N473 on the light chain. 26729242 4 0 theme first 673:677 arg1 lectin 679:684 The first lectin 669:684 The first lectin 669:684 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 2 1 theme Jacalin 333:339 arg1 enrichment 350:359 Jacalin affinity enrichment 333:359 Jacalin affinity enrichment 333:359 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 3 2 used used 598:601 arg2 workflow 584:591 a simpler and significantly faster new workflow 545:591 a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin 545:666 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 2 3 theme peptides 509:516 arg1 background 484:493 the high background 475:493 the high background of unmodified peptides 475:516 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 4 with protein- 368:375 arg1 chromatography 406:419 ERLIC chromatography 400:419 ERLIC chromatography 400:419 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 5 theme unmodified 498:507 arg1 peptides 509:516 unmodified peptides 498:516 unmodified peptides 498:516 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 6 theme lectin 260:265 arg1 Jacalin 267:273 the plant lectin Jacalin 250:273 the plant lectin Jacalin 250:273 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 4 7 theme general 695:701 arg1 enrichment 716:725 general glycopeptide enrichment 695:725 general glycopeptide enrichment 695:725 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 2 8 from % 208:208 arg1 serum 236:240 human serum 230:240 human serum 230:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 9 with peptide-level 381:393 arg1 chromatography 406:419 ERLIC chromatography 400:419 ERLIC chromatography 400:419 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 5 10 theme enriched 853:860 arg1 samples 862:868 enriched samples 853:868 enriched samples 853:868 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 2 11 theme plant 254:258 arg1 Jacalin 267:273 the plant lectin Jacalin 250:273 the plant lectin Jacalin 250:273 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 4 12 with glycopeptides 776:788 arg1 structures 810:819 Galβ1-3GalNAcα structures 795:819 Galβ1-3GalNAcα structures 795:819 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 4 13 gly glycopeptides 776:788 arg2 glycopeptides 776:788 O-linked glycopeptides 767:788 O-linked glycopeptides with Galβ1-3GalNAcα structures 767:819 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 2 14 theme further 426:432 arg1 step 445:448 a further enrichment step 424:448 a further enrichment step in between 424:459 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 1 15 theme human 180:184 arg1 serum 186:190 human serum 180:190 human serum 180:190 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 2 16 theme enrichment 434:443 arg1 step 445:448 a further enrichment step 424:448 a further enrichment step in between 424:459 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 17 theme high 479:482 arg1 background 484:493 the high background 475:493 the high background of unmodified peptides 475:516 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 18 from O-glycans 217:225 arg1 serum 236:240 human serum 230:240 human serum 230:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 5 19 theme new 886:888 arg1 method 909:914 the new sample preparation method 882:914 the new sample preparation method 882:914 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 5 19 theme new 886:888 arg1 selective 924:932 selective 924:932 selective 924:932 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 0 20 theme O-glycosylation 0:14 arg1 sites 16:20 O-glycosylation sites 0:20 O-glycosylation sites 0:20 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 6 21 theme glycosylation 987:999 arg1 sites 1001:1005 52 unique glycosylation sites 977:1005 52 unique glycosylation sites in 20 proteins 977:1020 Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. 26729242 3 22 theme simpler 547:553 arg1 workflow 584:591 a simpler and significantly faster new workflow 545:591 a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin 545:666 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 2 23 from serum 236:240 arg1 O-glycans 217:225 the O-glycans 213:225 the O-glycans in human serum 213:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 23 from serum 236:240 arg1 % 208:208 about 70 % 199:208 about 70 % of the O-glycans in human serum 199:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 3 24 theme wheat 629:633 arg1 agglutinin 640:649 wheat germ agglutinin 629:649 wheat germ agglutinin 629:649 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 5 25 theme spectrometric 827:839 arg1 analysis 841:848 Mass spectrometric analysis 822:848 Mass spectrometric analysis of enriched samples 822:868 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 5 26 theme Mass 822:825 arg1 analysis 841:848 Mass spectrometric analysis 822:848 Mass spectrometric analysis of enriched samples 822:868 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 4 27 gly glycopeptide 703:714 arg2 glycopeptide 703:714 general glycopeptide enrichment 695:725 general glycopeptide enrichment 695:725 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 6 28 gly glycosylation 987:999 arg2 sites 1001:1005 52 unique glycosylation sites 977:1005 52 unique glycosylation sites in 20 proteins 977:1020 Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. 26729242 6 28 gly glycosylation 987:999 arg2 52 977:978 52 977:978 52 977:978 Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. 26729242 3 29 theme faster 573:578 arg1 workflow 584:591 a simpler and significantly faster new workflow 545:591 a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin 545:666 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 4 30 theme Galβ1-3GalNAcα 795:808 arg1 structures 810:819 Galβ1-3GalNAcα structures 795:819 Galβ1-3GalNAcα structures 795:819 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 1 31 gly glycopeptides 110:122 arg2 glycopeptides 110:122 O-linked glycopeptides 101:122 O-linked glycopeptides bearing mucin core-1 type structures 101:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 1 32 theme O-linked 101:108 arg1 glycopeptides 110:122 O-linked glycopeptides 101:122 O-linked glycopeptides bearing mucin core-1 type structures 101:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 2 33 theme ERLIC 400:404 arg1 chromatography 406:419 ERLIC chromatography 400:419 ERLIC chromatography 400:419 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 0 34 theme core-1 44:49 arg1 glycopeptides 56:68 mucin core-1 type glycopeptides 38:68 mucin core-1 type glycopeptides from human serum 38:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 0 35 from serum 81:85 arg1 glycopeptides 56:68 mucin core-1 type glycopeptides 38:68 mucin core-1 type glycopeptides from human serum 38:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 0 36 gly O-glycosylation 0:14 arg2 sites 16:20 O-glycosylation sites 0:20 O-glycosylation sites 0:20 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 5 37 theme sample 890:895 arg1 method 909:914 the new sample preparation method 882:914 the new sample preparation method 882:914 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 5 37 theme sample 890:895 arg1 selective 924:932 selective 924:932 selective 924:932 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 5 38 theme samples 862:868 arg1 analysis 841:848 Mass spectrometric analysis 822:848 Mass spectrometric analysis of enriched samples 822:868 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 0 39 theme mucin 38:42 arg1 glycopeptides 56:68 mucin core-1 type glycopeptides 38:68 mucin core-1 type glycopeptides from human serum 38:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 2 40 dep protein- 368:375 arg1 the 364:366 the 364:366 the 364:366 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 41 theme O-glycans 217:225 arg1 O-glycans 217:225 the O-glycans 213:225 the O-glycans in human serum 213:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 2 41 theme O-glycans 217:225 arg1 % 208:208 about 70 % 199:208 about 70 % of the O-glycans in human serum 199:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 5 42 theme preparation 897:907 arg1 method 909:914 the new sample preparation method 882:914 the new sample preparation method 882:914 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 5 42 theme preparation 897:907 arg1 selective 924:932 selective 924:932 selective 924:932 Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. 26729242 6 43 theme unique 980:985 arg1 sites 1001:1005 52 unique glycosylation sites 977:1005 52 unique glycosylation sites in 20 proteins 977:1020 Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. 26729242 4 44 link O-linked 767:774 arg1 glycopeptides 776:788 O-linked glycopeptides 767:788 O-linked glycopeptides with Galβ1-3GalNAcα structures 767:819 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 0 45 theme type 51:54 arg1 glycopeptides 56:68 mucin core-1 type glycopeptides 38:68 mucin core-1 type glycopeptides from human serum 38:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 6 46 from sites 1001:1005 arg1 proteins 1013:1020 20 proteins 1010:1020 20 proteins 1010:1020 Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. 26729242 4 47 theme glycopeptide 703:714 arg1 enrichment 716:725 general glycopeptide enrichment 695:725 general glycopeptide enrichment 695:725 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 2 48 theme human 230:234 arg1 serum 236:240 human serum 230:240 human serum 230:240 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 3 49 theme new 580:582 arg1 workflow 584:591 a simpler and significantly faster new workflow 545:591 a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin 545:666 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 1 50 theme mucin 132:136 arg1 structures 150:159 mucin core-1 type structures 132:159 mucin core-1 type structures 132:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 4 51 theme O-linked 767:774 arg1 glycopeptides 776:788 O-linked glycopeptides 767:788 O-linked glycopeptides with Galβ1-3GalNAcα structures 767:819 The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. 26729242 3 52 theme germ 635:638 arg1 agglutinin 640:649 wheat germ agglutinin 629:649 wheat germ agglutinin 629:649 In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. 26729242 1 53 theme core-1 138:143 arg1 structures 150:159 mucin core-1 type structures 132:159 mucin core-1 type structures 132:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 2 54 theme successful 299:308 arg1 protocol 310:317 a previously successful protocol 286:317 a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides 286:516 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 26729242 0 55 theme human 75:79 arg1 serum 81:85 human serum 75:85 human serum 75:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 1 56 theme type 145:148 arg1 structures 150:159 mucin core-1 type structures 132:159 mucin core-1 type structures 132:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 1 57 link O-linked 101:108 arg1 glycopeptides 110:122 O-linked glycopeptides 101:122 O-linked glycopeptides bearing mucin core-1 type structures 101:159 In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. 26729242 0 58 gly glycopeptides 56:68 arg2 glycopeptides 56:68 mucin core-1 type glycopeptides 38:68 mucin core-1 type glycopeptides from human serum 38:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 0 58 gly glycopeptides 56:68 arg1 serum 81:85 human serum 75:85 human serum 75:85 O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. 26729242 2 59 theme affinity 341:348 arg1 enrichment 350:359 Jacalin affinity enrichment 333:359 Jacalin affinity enrichment 333:359 Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. 24308486 7 0 theme terminal 1217:1224 arg1 acids 1245:1249 one or two terminal N-acetylneuraminic acids 1206:1249 one or two terminal N-acetylneuraminic acids 1206:1249 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 0 1 theme immune 160:165 arg1 reactions 167:175 antiparasitic and allergic immune reactions 133:175 antiparasitic and allergic immune reactions 133:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 7 2 theme nonmyeloma 1108:1117 arg1 IgE 1119:1121 the nonmyeloma IgE 1104:1121 the nonmyeloma IgE 1104:1121 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 6 3 theme complex-type 1078:1089 arg1 glycans 1091:1097 complex-type glycans 1078:1097 complex-type glycans 1078:1097 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 7 4 theme N-acetylneuraminic 1226:1243 arg1 acids 1245:1249 one or two terminal N-acetylneuraminic acids 1206:1249 one or two terminal N-acetylneuraminic acids 1206:1249 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 2 5 theme donors 513:518 arg1 serum 483:487 the pooled serum 472:487 the pooled serum of multiple nondiseased donors 472:518 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 6 6 contain contained 922:930 arg1 samples 901:907 all three samples 891:907 all three samples 891:907 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 6 6 contain contained 922:930 arg1 Asn275 915:920 site Asn275 910:920 site Asn275 910:920 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 6 6 contain contained 922:930 arg2 structures 960:969 exclusively oligomannosidic structures 932:969 exclusively oligomannosidic structures 932:969 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 6 6 contain contained 922:930 arg2 Asn275 915:920 site Asn275 910:920 site Asn275 910:920 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 8 7 theme glycan 1338:1343 arg1 structures 1345:1354 triantennary and tetraantennary glycan structures 1306:1354 structures 1345:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 8 8 theme structures 1345:1354 arg1 abundance 1366:1374 a low abundance 1360:1374 a low abundance of species with a bisecting N-acetylglucosamine 1360:1422 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 8 8 theme structures 1345:1354 arg1 abundance 1293:1301 a higher abundance 1284:1301 a higher abundance of triantennary and tetraantennary glycan structures 1284:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 8 9 theme tetraantennary 1323:1336 arg1 structures 1345:1354 triantennary and tetraantennary glycan structures 1306:1354 structures 1345:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 3 10 gly N-glycosylation 723:737 arg2 seven 707:711 seven 707:711 seven 707:711 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 3 10 gly N-glycosylation 723:737 arg2 sites 739:743 all seven potential N-glycosylation sites 703:743 all seven potential N-glycosylation sites 703:743 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 1 11 theme IgE 178:180 arg1 glycosylation 182:194 IgE glycosylation 178:194 IgE glycosylation 178:194 IgE glycosylation is known to exhibit significant interindividual variation, and several reports have indicated its relevance in determining IgE activity. 24308486 0 12 theme heterodimeric 94:106 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) 0:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 12 theme heterodimeric 94:106 arg1 glycoprotein 108:119 a heterodimeric glycoprotein 92:119 a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions 92:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 2 13 theme pooled 476:481 arg1 serum 483:487 the pooled serum 472:487 the pooled serum of multiple nondiseased donors 472:518 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 0 14 theme Site-specific 0:12 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) 0:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 14 theme Site-specific 0:12 arg1 glycoprotein 108:119 a heterodimeric glycoprotein 92:119 a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions 92:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 8 15 with species 1379:1385 arg1 N-acetylglucosamine 1404:1422 a bisecting N-acetylglucosamine 1392:1422 a bisecting N-acetylglucosamine 1392:1422 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 0 16 theme N-glycosylation 14:28 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) 0:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 16 theme N-glycosylation 14:28 arg1 glycoprotein 108:119 a heterodimeric glycoprotein 92:119 a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions 92:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 3 17 theme potential 713:721 arg1 sites 739:743 all seven potential N-glycosylation sites 703:743 all seven potential N-glycosylation sites 703:743 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 8 18 theme species 1379:1385 arg1 abundance 1366:1374 a low abundance 1360:1374 a low abundance of species with a bisecting N-acetylglucosamine 1360:1422 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 8 18 theme species 1379:1385 arg1 abundance 1293:1301 a higher abundance 1284:1301 a higher abundance of triantennary and tetraantennary glycan structures 1284:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 8 19 theme low 1362:1364 arg1 abundance 1366:1374 a low abundance 1360:1374 a low abundance of species with a bisecting N-acetylglucosamine 1360:1422 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 0 20 theme allergic 151:158 arg1 reactions 167:175 antiparasitic and allergic immune reactions 133:175 antiparasitic and allergic immune reactions 133:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 2 21 theme different 405:413 arg1 sources 415:421 three different sources 399:421 three different sources: IgE from the serum of a hyperimmune donor 399:464 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 22 theme hyperimmune 448:458 arg1 donor 460:464 a hyperimmune donor 446:464 a hyperimmune donor 446:464 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 3 23 theme N-glycosylation 723:737 arg1 sites 739:743 all seven potential N-glycosylation sites 703:743 all seven potential N-glycosylation sites 703:743 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 3 24 theme sites 739:743 arg1 coverage 691:698 coverage 691:698 coverage of all seven potential N-glycosylation sites 691:743 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 8 25 theme triantennary 1306:1317 arg1 structures 1345:1354 triantennary and tetraantennary glycan structures 1306:1354 structures 1345:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 9 26 theme glycosylation 1463:1475 arg1 comparison 1445:1454 comparison 1445:1454 comparison of the glycosylation of IgE samples 1445:1490 Our approach allows comparison of the glycosylation of IgE samples in a site-specific manner. 24308486 9 27 theme IgE 1480:1482 arg1 samples 1484:1490 IgE samples 1480:1490 IgE samples 1480:1490 Our approach allows comparison of the glycosylation of IgE samples in a site-specific manner. 24308486 6 28 dep contained 922:930 arg1 whereas 1002:1008 whereas 1002:1008 whereas 1002:1008 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 0 29 gly glycoprotein 108:119 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) 0:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 29 gly glycoprotein 108:119 arg1 glycoprotein 108:119 a heterodimeric glycoprotein 92:119 a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions 92:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 30 theme immunoglobulin 48:61 arg1 IgE 84:86 IgE 84:86 IgE 84:86 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 30 theme immunoglobulin 48:61 arg1 E 81:81 human immunoglobulin e. Immunoglobulin E 42:81 human immunoglobulin e. Immunoglobulin E (IgE) 42:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 1 31 theme IgE 319:321 arg1 activity 323:330 IgE activity 319:330 IgE activity 319:330 IgE glycosylation is known to exhibit significant interindividual variation, and several reports have indicated its relevance in determining IgE activity. 24308486 2 32 theme pooled 534:539 arg1 serum 541:545 the pooled serum 530:545 the pooled serum of 2 patients with IgE myeloma 530:576 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 5 33 theme Site 848:851 arg1 Asn264 853:858 Site Asn264 848:858 Site Asn264 848:858 Site Asn264 was shown to be unoccupied. 24308486 1 34 theme significant 216:226 arg1 variation 244:252 significant interindividual variation 216:252 significant interindividual variation 216:252 IgE glycosylation is known to exhibit significant interindividual variation, and several reports have indicated its relevance in determining IgE activity. 24308486 0 35 theme human 42:46 arg1 IgE 84:86 IgE 84:86 IgE 84:86 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 35 theme human 42:46 arg1 E 81:81 human immunoglobulin e. Immunoglobulin E 42:81 human immunoglobulin e. Immunoglobulin E (IgE) 42:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 6 36 theme oligomannosidic 944:958 arg1 structures 960:969 exclusively oligomannosidic structures 932:969 exclusively oligomannosidic structures 932:969 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 0 37 theme antiparasitic 133:145 arg1 reactions 167:175 antiparasitic and allergic immune reactions 133:175 antiparasitic and allergic immune reactions 133:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 8 38 theme myeloma 1265:1271 arg1 IgE 1273:1275 myeloma IgE 1265:1275 myeloma IgE 1265:1275 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 0 39 gly N-glycosylation 14:28 arg1 IgE 84:86 IgE 84:86 IgE 84:86 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 39 gly N-glycosylation 14:28 arg1 E 81:81 human immunoglobulin e. Immunoglobulin E 42:81 human immunoglobulin e. Immunoglobulin E (IgE) 42:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 1 40 theme interindividual 228:242 arg1 variation 244:252 significant interindividual variation 216:252 significant interindividual variation 216:252 IgE glycosylation is known to exhibit significant interindividual variation, and several reports have indicated its relevance in determining IgE activity. 24308486 0 41 theme Immunoglobulin 66:79 arg1 IgE 84:86 IgE 84:86 IgE 84:86 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 41 theme Immunoglobulin 66:79 arg1 E 81:81 human immunoglobulin e. Immunoglobulin E 42:81 human immunoglobulin e. Immunoglobulin E (IgE) 42:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 2 42 theme IgE 566:568 arg1 myeloma 570:576 IgE myeloma 566:576 IgE myeloma 566:576 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 43 theme multiple 492:499 arg1 donors 513:518 multiple nondiseased donors 492:518 multiple nondiseased donors 492:518 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 8 44 theme bisecting 1394:1402 arg1 N-acetylglucosamine 1404:1422 a bisecting N-acetylglucosamine 1392:1422 a bisecting N-acetylglucosamine 1392:1422 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 2 45 theme glycosylation 364:376 arg1 analysis 378:385 site-specific glycosylation analysis 350:385 site-specific glycosylation analysis of IgE 350:392 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 0 46 theme e. 63:64 arg1 IgE 84:86 IgE 84:86 IgE 84:86 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 46 theme e. 63:64 arg1 E 81:81 human immunoglobulin e. Immunoglobulin E 42:81 human immunoglobulin e. Immunoglobulin E (IgE) 42:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 2 47 gly glycosylation 364:376 arg1 IgE 390:392 IgE 390:392 IgE 390:392 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 6 48 contain contained 1056:1064 arg1 sites 1010:1014 sites Asn21, Asn49, Asn99, Asn146, and Asn252 1010:1054 sites Asn21, Asn49, Asn99, Asn146, and Asn252 1010:1054 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 6 48 contain contained 1056:1064 arg2 glycans 1091:1097 complex-type glycans 1078:1097 complex-type glycans 1078:1097 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 2 49 theme site-specific 350:362 arg1 analysis 378:385 site-specific glycosylation analysis 350:385 site-specific glycosylation analysis of IgE 350:392 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 50 theme patients 552:559 arg1 serum 541:545 the pooled serum 530:545 the pooled serum of 2 patients with IgE myeloma 530:576 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 9 51 gly glycosylation 1463:1475 arg1 samples 1484:1490 IgE samples 1480:1490 IgE samples 1480:1490 Our approach allows comparison of the glycosylation of IgE samples in a site-specific manner. 24308486 3 52 theme heavy 583:587 arg1 chains 589:594 The heavy chains 579:594 The heavy chains 579:594 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 8 53 theme higher 1286:1291 arg1 abundance 1293:1301 a higher abundance 1284:1301 a higher abundance of triantennary and tetraantennary glycan structures 1284:1354 In contrast, myeloma IgE showed a higher abundance of triantennary and tetraantennary glycan structures and a low abundance of species with a bisecting N-acetylglucosamine. 24308486 4 54 theme resulting 750:758 arg1 peptides 768:775 The resulting (glyco-)peptides 746:775 The resulting (glyco-)peptides 746:775 The resulting (glyco-)peptides were analyzed by nano-reversed-phase-LC-MS/MS and MALDI-TOF/TOF-MS/MS. 24308486 4 55 theme glyco- 761:766 arg1 peptides 768:775 The resulting (glyco-)peptides 746:775 The resulting (glyco-)peptides 746:775 The resulting (glyco-)peptides were analyzed by nano-reversed-phase-LC-MS/MS and MALDI-TOF/TOF-MS/MS. 24308486 2 56 theme IgE 390:392 arg1 analysis 378:385 site-specific glycosylation analysis 350:385 site-specific glycosylation analysis of IgE 350:392 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 9 57 theme site-specific 1497:1509 arg1 manner 1511:1516 a site-specific manner 1495:1516 a site-specific manner 1495:1516 Our approach allows comparison of the glycosylation of IgE samples in a site-specific manner. 24308486 2 58 theme nondiseased 501:511 arg1 donors 513:518 multiple nondiseased donors 492:518 multiple nondiseased donors 492:518 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 3 59 theme proteinase 644:653 arg1 K 655:655 proteinase K 644:655 proteinase K 644:655 The heavy chains were isolated and digested with either trypsin, proteinase K, or chymotrypsin, which permitted coverage of all seven potential N-glycosylation sites. 24308486 7 60 contain contained 1196:1204 arg1 biantennary 1159:1169 biantennary 1159:1169 biantennary 1159:1169 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 7 60 contain contained 1196:1204 arg1 majority 1128:1135 the majority 1124:1135 the majority of these glycans 1124:1152 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 7 60 contain contained 1196:1204 arg2 acids 1245:1249 one or two terminal N-acetylneuraminic acids 1206:1249 one or two terminal N-acetylneuraminic acids 1206:1249 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 7 61 theme glycans 1146:1152 arg1 biantennary 1159:1169 biantennary 1159:1169 biantennary 1159:1169 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 7 61 theme glycans 1146:1152 arg1 majority 1128:1135 the majority 1124:1135 the majority of these glycans 1124:1152 For the nonmyeloma IgE, the majority of these glycans were biantennary and core-fucosylated and contained one or two terminal N-acetylneuraminic acids. 24308486 9 62 theme samples 1484:1490 arg1 glycosylation 1463:1475 the glycosylation 1459:1475 the glycosylation of IgE samples 1459:1490 Our approach allows comparison of the glycosylation of IgE samples in a site-specific manner. 24308486 0 63 theme E 81:81 arg1 analysis 30:37 Site-specific N-glycosylation analysis 0:37 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) 0:87 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 0 63 theme E 81:81 arg1 glycoprotein 108:119 a heterodimeric glycoprotein 92:119 a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions 92:175 Site-specific N-glycosylation analysis of human immunoglobulin e. Immunoglobulin E (IgE) is a heterodimeric glycoprotein involved in antiparasitic and allergic immune reactions. 24308486 6 64 theme site 910:913 arg1 Asn275 915:920 site Asn275 910:920 site Asn275 910:920 In all three samples, site Asn275 contained exclusively oligomannosidic structures with between 2 and 9 mannoses, whereas sites Asn21, Asn49, Asn99, Asn146, and Asn252 contained exclusively complex-type glycans. 24308486 2 65 dep sources 415:421 arg1 IgE 424:426 IgE 424:426 three different sources: IgE from the serum of a hyperimmune donor 399:464 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 66 from serum 437:441 arg1 IgE 424:426 IgE 424:426 three different sources: IgE from the serum of a hyperimmune donor 399:464 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 67 theme donor 460:464 arg1 serum 437:441 the serum 433:441 the serum of a hyperimmune donor 433:464 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 2 68 with patients 552:559 arg1 myeloma 570:576 IgE myeloma 566:576 IgE myeloma 566:576 Here, we present site-specific glycosylation analysis of IgE from three different sources: IgE from the serum of a hyperimmune donor, from the pooled serum of multiple nondiseased donors, and from the pooled serum of 2 patients with IgE myeloma. 24308486 1 69 theme several 259:265 arg1 reports 267:273 several reports 259:273 several reports 259:273 IgE glycosylation is known to exhibit significant interindividual variation, and several reports have indicated its relevance in determining IgE activity. 26991339 0 0 theme proline-rich 73:84 arg1 protein 86:92 human basic salivary proline-rich protein 3M 52:95 the human basic salivary proline-rich protein 3M 48:95 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 1 1 theme fraction 288:295 arg1 mixture 151:157 the heterogeneous mixture 133:157 the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 133:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 1 theme fraction 288:295 arg1 component 256:264 a major component 248:264 a major component of the acidic soluble fraction of human whole saliva in the first years of life 248:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 2 dep analysis 464:471 arg1 sequencing 592:601 sequencing 592:601 sequencing 592:601 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 3 theme O-glycosylation 753:767 arg1 characterization 683:698 the structural characterization 668:698 the structural characterization of the peptide backbone 668:722 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 3 theme O-glycosylation 753:767 arg1 identification 728:741 identification 728:741 identification of N- and O-glycosylation sites 728:773 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 4 theme different 929:937 arg1 glycans 939:945 33 different glycans 926:945 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 926:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 3 5 theme sialylated 877:886 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 0 6 theme salivary 64:71 arg1 protein 86:92 human basic salivary proline-rich protein 3M 52:95 the human basic salivary proline-rich protein 3M 48:95 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 3 7 theme glycans 888:894 arg1 combination 838:848 the combination 834:848 the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 834:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 8 theme human 300:304 arg1 saliva 312:317 human whole saliva 300:317 human whole saliva 300:317 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 9 gly glycoforms 162:171 arg1 protein 208:214 the basic salivary proline-rich protein 3M 176:217 the basic salivary proline-rich protein 3M 176:217 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 0 10 link O-linked 7:14 arg1 site 30:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 1 11 theme PRB3-M 231:236 arg1 locus 238:242 PRB3-M locus 231:242 PRB3-M locus 231:242 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 12 theme whole 306:310 arg1 saliva 312:317 human whole saliva 300:317 human whole saliva 300:317 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 13 with chromatography 386:399 arg1 spectrometry 579:590 tandem mass spectrometry 567:590 tandem mass spectrometry 567:590 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 13 with chromatography 386:399 arg1 analysis 464:471 high-resolution electrospray ionization mass spectrometry analysis 406:471 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 14 theme structural 672:681 arg1 characterization 683:698 the structural characterization 668:698 the structural characterization of the peptide backbone 668:722 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 15 theme N-linked 947:954 arg1 glycans 939:945 33 different glycans 926:945 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 926:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 16 theme saliva 312:317 arg1 fraction 288:295 the acidic soluble fraction 269:295 the acidic soluble fraction of human whole saliva in the first years of life 269:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 3 17 theme different 855:863 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 3 18 theme Asparagine 959:968 arg1 residues 970:977 Asparagine residues 959:977 Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 959:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 19 theme heterogeneous 137:149 arg1 mixture 151:157 the heterogeneous mixture 133:157 the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 133:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 19 theme heterogeneous 137:149 arg1 component 256:264 a major component 248:264 a major component of the acidic soluble fraction of human whole saliva in the first years of life 248:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 20 from years 332:336 arg1 mixture 151:157 the heterogeneous mixture 133:157 the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 133:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 20 from years 332:336 arg1 component 256:264 a major component 248:264 a major component of the acidic soluble fraction of human whole saliva in the first years of life 248:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 21 theme high-resolution 406:420 arg1 spectrometry 451:462 high-resolution electrospray ionization mass spectrometry 406:462 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 22 theme glycans 939:945 arg1 combination 838:848 the combination 834:848 the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 834:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 2 23 theme electrospray 422:433 arg1 spectrometry 451:462 high-resolution electrospray ionization mass spectrometry 406:462 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 24 theme GluC 645:648 arg1 digestion 650:658 Endoproteinase GluC digestion 630:658 Endoproteinase GluC digestion 630:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 25 theme backbone 715:722 arg1 characterization 683:698 the structural characterization 668:698 the structural characterization of the peptide backbone 668:722 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 25 theme backbone 715:722 arg1 identification 728:741 identification 728:741 identification of N- and O-glycosylation sites 728:773 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 26 link N-linked 947:954 arg1 glycans 939:945 33 different glycans 926:945 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 926:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 0 27 theme N- 0:1 arg1 site 30:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 2 28 theme Endoproteinase 630:643 arg1 digestion 650:658 Endoproteinase GluC digestion 630:658 Endoproteinase GluC digestion 630:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 29 theme peptide 707:713 arg1 backbone 715:722 the peptide backbone 703:722 the peptide backbone 703:722 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 30 link O-linked 896:903 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 3 31 gly sialylated 877:886 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 32 theme glycoforms 162:171 arg1 mixture 151:157 the heterogeneous mixture 133:157 the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 133:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 32 theme glycoforms 162:171 arg1 component 256:264 a major component 248:264 a major component of the acidic soluble fraction of human whole saliva in the first years of life 248:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 0 33 theme glycosylation 16:28 arg1 site 30:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 2 34 with N-deglycosylation 516:532 arg1 F 561:561 Peptide-N-Glycosidase F 539:561 Peptide-N-Glycosidase F 539:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 35 theme neutral 865:871 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 36 from component 256:264 arg1 years 332:336 the first years 322:336 the first years of life 322:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 0 37 theme O-linked 7:14 arg1 site 30:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 3 38 theme heterogeneous 780:792 arg1 mixture 794:800 The heterogeneous mixture 776:800 The heterogeneous mixture of the proteoforms 776:819 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 39 theme first 326:330 arg1 years 332:336 the first years 322:336 the first years of life 322:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 40 theme high-performance 362:377 arg1 chromatography 386:399 Reversed-phase high-performance liquid chromatography 347:399 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion 347:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 41 theme major 250:254 arg1 mixture 151:157 the heterogeneous mixture 133:157 the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 133:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 41 theme major 250:254 arg1 component 256:264 a major component 248:264 a major component of the acidic soluble fraction of human whole saliva in the first years of life 248:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 42 theme Peptide-N-Glycosidase 539:559 arg1 F 561:561 Peptide-N-Glycosidase F 539:561 Peptide-N-Glycosidase F 539:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 43 theme Reversed-phase 347:360 arg1 chromatography 386:399 Reversed-phase high-performance liquid chromatography 347:399 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion 347:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 44 dep N- 746:747 arg1 sites 769:773 sites 769:773 sites 769:773 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 45 theme basic 180:184 arg1 protein 208:214 the basic salivary proline-rich protein 3M 176:217 the basic salivary proline-rich protein 3M 176:217 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 46 theme spectrometry 451:462 arg1 analysis 464:471 high-resolution electrospray ionization mass spectrometry analysis 406:471 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 47 theme salivary 186:193 arg1 protein 208:214 the basic salivary proline-rich protein 3M 176:217 the basic salivary proline-rich protein 3M 176:217 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 48 theme mass 446:449 arg1 spectrometry 451:462 high-resolution electrospray ionization mass spectrometry 406:462 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 49 theme proteoforms 487:497 arg1 spectrometry 579:590 tandem mass spectrometry 567:590 tandem mass spectrometry 567:590 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 49 theme proteoforms 487:497 arg1 analysis 464:471 high-resolution electrospray ionization mass spectrometry analysis 406:471 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 50 theme life 341:344 arg1 years 332:336 the first years 322:336 the first years of life 322:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 3 51 from positions 982:990 arg1 residues 970:977 Asparagine residues 959:977 Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213 959:1031 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 2 52 theme N- 746:747 arg1 characterization 683:698 the structural characterization 668:698 the structural characterization of the peptide backbone 668:722 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 52 theme N- 746:747 arg1 identification 728:741 identification 728:741 identification of N- and O-glycosylation sites 728:773 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 53 from fraction 288:295 arg1 years 332:336 the first years 322:336 the first years of life 322:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 3 54 theme proteoforms 809:819 arg1 mixture 794:800 The heterogeneous mixture 776:800 The heterogeneous mixture of the proteoforms 776:819 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 2 55 theme ionization 435:444 arg1 spectrometry 451:462 high-resolution electrospray ionization mass spectrometry 406:462 high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 406:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 0 56 gly glycosylation 16:28 arg2 site 30:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site 0:33 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 1 57 theme present 105:111 arg1 study 113:117 the present study 101:117 the present study 101:117 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 1 58 theme acidic 273:278 arg1 fraction 288:295 the acidic soluble fraction 269:295 the acidic soluble fraction of human whole saliva in the first years of life 269:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 59 theme mass 574:577 arg1 spectrometry 579:590 tandem mass spectrometry 567:590 tandem mass spectrometry 567:590 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 60 theme peptides 606:613 arg1 chromatography 386:399 Reversed-phase high-performance liquid chromatography 347:399 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion 347:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 1 61 theme proline-rich 195:206 arg1 protein 208:214 the basic salivary proline-rich protein 3M 176:217 the basic salivary proline-rich protein 3M 176:217 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 2 62 theme tandem 567:572 arg1 spectrometry 579:590 tandem mass spectrometry 567:590 tandem mass spectrometry 567:590 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 2 63 theme intact 480:485 arg1 proteoforms 487:497 the intact proteoforms 476:497 the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F 476:561 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 3 64 theme O-linked 896:903 arg1 glycans 888:894 8 different neutral and sialylated glycans 853:894 8 different neutral and sialylated glycans O-linked to Threonine 50 853:919 The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213. 26991339 1 65 theme protein 208:214 arg1 glycoforms 162:171 glycoforms 162:171 glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, 162:243 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 26991339 0 66 theme basic 58:62 arg1 protein 86:92 human basic salivary proline-rich protein 3M 52:95 the human basic salivary proline-rich protein 3M 48:95 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 2 67 theme liquid 379:384 arg1 chromatography 386:399 Reversed-phase high-performance liquid chromatography 347:399 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion 347:658 Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. 26991339 0 68 theme human 52:56 arg1 protein 86:92 human basic salivary proline-rich protein 3M 52:95 the human basic salivary proline-rich protein 3M 48:95 N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M. 26991339 1 69 theme soluble 280:286 arg1 fraction 288:295 the acidic soluble fraction 269:295 the acidic soluble fraction of human whole saliva in the first years of life 269:344 In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. 25995273 7 0 from proteins 1143:1150 arg1 proteins 1094:1101 34 proteins 1091:1101 34 proteins 1091:1101 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 7 0 from proteins 1143:1150 arg1 sites 1126:1130 13 O-glycosylation sites 1107:1130 13 O-glycosylation sites 1107:1130 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 3 1 theme reticulum 536:544 arg1 structures 489:498 The N-glycan structures 476:498 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins 476:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 1 2 from enrichment 87:96 arg1 liver 165:169 mouse liver 159:169 mouse liver 159:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 7 3 from sites 1126:1130 arg1 data 1053:1056 comparative brain/liver glycosylation data 1015:1056 comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins 1015:1150 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 2 4 gly glycopeptide 437:448 arg2 glycopeptide 437:448 glycopeptide analysis 437:457 glycopeptide analysis 437:457 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 7 5 theme comparative 1015:1025 arg1 data 1053:1056 comparative brain/liver glycosylation data 1015:1056 comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins 1015:1150 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 3 6 theme N-glycan 480:487 arg1 structures 489:498 The N-glycan structures 476:498 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins 476:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 7 7 theme brain/liver 1027:1037 arg1 data 1053:1056 comparative brain/liver glycosylation data 1015:1056 comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins 1015:1150 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 6 8 dep no 979:980 arg1 overlap 982:988 overlap 982:988 overlap 982:988 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 3 9 theme proteins 579:586 arg1 structures 489:498 The N-glycan structures 476:498 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins 476:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 3 10 theme transmembrane 565:577 arg1 proteins 579:586 transmembrane proteins 565:586 transmembrane proteins 565:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 5 11 gly glycopeptides 755:767 arg2 glycopeptides 755:767 several mouse liver and mouse brain glycopeptides 719:767 several mouse liver and mouse brain glycopeptides 719:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 1 12 from liver 165:169 arg1 glycopeptides 140:152 intact glycopeptides 133:152 intact glycopeptides from mouse liver 133:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 12 from liver 165:169 arg1 enrichment 87:96 enrichment 87:96 enrichment 87:96 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 12 from liver 165:169 arg1 analysis 121:128 mass spectrometric analysis 102:128 mass spectrometric analysis 102:128 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 2 13 theme electron 402:409 arg1 dissociation 420:431 electron transfer dissociation 402:431 electron transfer dissociation 402:431 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 5 14 gly glycosylation 693:705 arg1 glycopeptides 755:767 several mouse liver and mouse brain glycopeptides 719:767 several mouse liver and mouse brain glycopeptides 719:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 2 15 theme variants 287:294 arg1 Incorporation 248:260 Incorporation 248:260 Incorporation of different sialic acid variants in both N- and O-linked glycans 248:326 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 2 16 from Incorporation 248:260 arg1 N- 304:305 N- 304:305 N- 304:305 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 2 16 from Incorporation 248:260 arg1 glycans 320:326 O-linked glycans 311:326 O-linked glycans 311:326 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 1 17 theme site-specific 186:198 arg1 data 223:226 site-specific N- and O-glycosylation data 186:226 site-specific N- and O-glycosylation data for ∼ 130 proteins 186:245 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 7 18 theme O-glycosylation 1110:1124 arg1 sites 1126:1130 13 O-glycosylation sites 1107:1130 13 O-glycosylation sites 1107:1130 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 1 19 theme N- 200:201 arg1 data 223:226 site-specific N- and O-glycosylation data 186:226 site-specific N- and O-glycosylation data for ∼ 130 proteins 186:245 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 0 20 theme Tissue-Specific 0:14 arg1 Glycosylation 16:28 Tissue-Specific Glycosylation 0:28 Tissue-Specific Glycosylation 0:28 Tissue-Specific Glycosylation at the Glycopeptide Level. 25995273 1 21 theme mass 102:105 arg1 analysis 121:128 mass spectrometric analysis 102:128 mass spectrometric analysis 102:128 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 2 22 theme collisional 375:385 arg1 activation 387:396 collisional activation 375:396 collisional activation 375:396 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 2 23 theme different 265:273 arg1 variants 287:294 different sialic acid variants 265:294 different sialic acid variants 265:294 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 5 24 theme mouse 743:747 arg1 brain 749:753 mouse brain 743:753 mouse brain 743:753 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 6 25 theme same 868:871 arg1 protein 873:879 the same protein 864:879 the same protein 864:879 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 1 26 theme spectrometric 107:119 arg1 analysis 121:128 mass spectrometric analysis 102:128 mass spectrometric analysis 102:128 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 2 27 theme glycopeptide 437:448 arg1 analysis 450:457 glycopeptide analysis 437:457 glycopeptide analysis 437:457 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 6 28 theme Tissue-specific 784:798 arg1 differences 800:810 Tissue-specific differences 784:810 Tissue-specific differences in glycosylation 784:827 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 5 29 theme brain 749:753 arg1 glycopeptides 755:767 several mouse liver and mouse brain glycopeptides 719:767 several mouse liver and mouse brain glycopeptides 719:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 6 30 theme structures 932:941 arg1 spectrum 913:920 a similar spectrum 903:920 a similar spectrum of glycan structures 903:941 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 3 31 theme endoplasmic 524:534 arg1 ER 547:548 ER 547:548 ER 547:548 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 3 31 theme endoplasmic 524:534 arg1 reticulum 536:544 endoplasmic reticulum 524:544 endoplasmic reticulum (ER) 524:549 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 5 32 theme glycosylation 693:705 arg1 patterns 707:714 The glycosylation patterns 689:714 The glycosylation patterns of several mouse liver and mouse brain glycopeptides 689:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 1 33 dep enrichment 87:96 arg1 the 83:85 the 83:85 the 83:85 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 5 34 theme glycopeptides 755:767 arg1 patterns 707:714 The glycosylation patterns 689:714 The glycosylation patterns of several mouse liver and mouse brain glycopeptides 689:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 6 35 theme glycan 925:930 arg1 structures 932:941 glycan structures 925:941 glycan structures 925:941 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 0 36 theme Glycopeptide 37:48 arg1 Level 50:54 the Glycopeptide Level 33:54 the Glycopeptide Level 33:54 Tissue-Specific Glycosylation at the Glycopeptide Level. 25995273 2 37 link O-linked 311:318 arg1 glycans 320:326 O-linked glycans 311:326 O-linked glycans 311:326 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 1 38 gly glycopeptides 140:152 arg1 liver 165:169 mouse liver 159:169 mouse liver 159:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 38 gly glycopeptides 140:152 arg2 glycopeptides 140:152 intact glycopeptides 133:152 intact glycopeptides from mouse liver 133:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 2 39 theme transfer 411:418 arg1 dissociation 420:431 electron transfer dissociation 402:431 electron transfer dissociation 402:431 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 2 40 theme O-linked 311:318 arg1 glycans 320:326 O-linked glycans 311:326 O-linked glycans 311:326 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 5 41 theme several 719:725 arg1 glycopeptides 755:767 several mouse liver and mouse brain glycopeptides 719:767 several mouse liver and mouse brain glycopeptides 719:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 6 42 from differences 800:810 arg1 glycosylation 815:827 glycosylation 815:827 glycosylation 815:827 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 7 43 theme 34 1091:1092 arg1 proteins 1094:1101 34 proteins 1091:1101 34 proteins 1091:1101 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 7 44 gly O-glycosylation 1110:1124 arg2 sites 1126:1130 13 O-glycosylation sites 1107:1130 13 O-glycosylation sites 1107:1130 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 7 44 gly O-glycosylation 1110:1124 arg2 13 1107:1108 13 1107:1108 13 1107:1108 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 5 45 theme mouse 727:731 arg1 liver 733:737 mouse liver 727:737 mouse liver 727:737 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 6 46 theme similar 905:911 arg1 spectrum 913:920 a similar spectrum 903:920 a similar spectrum of glycan structures 903:941 Tissue-specific differences in glycosylation were observed between sites within the same protein: Some sites displayed a similar spectrum of glycan structures in both tissues, whereas for others no overlap was observed. 25995273 7 47 theme N-glycosylation 1064:1078 arg1 sites 1080:1084 50 N-glycosylation sites 1061:1084 50 N-glycosylation sites 1061:1084 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 1 48 theme intact 133:138 arg1 glycopeptides 140:152 intact glycopeptides 133:152 intact glycopeptides from mouse liver 133:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 7 49 gly N-glycosylation 1064:1078 arg2 sites 1080:1084 50 N-glycosylation sites 1061:1084 50 N-glycosylation sites 1061:1084 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 7 49 gly N-glycosylation 1064:1078 arg2 50 1061:1062 50 1061:1062 50 1061:1062 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 5 50 theme liver 733:737 arg1 glycopeptides 755:767 several mouse liver and mouse brain glycopeptides 719:767 several mouse liver and mouse brain glycopeptides 719:767 The glycosylation patterns of several mouse liver and mouse brain glycopeptides were compared. 25995273 2 51 theme acid 282:285 arg1 variants 287:294 different sialic acid variants 265:294 different sialic acid variants 265:294 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 1 52 theme glycopeptides 140:152 arg1 enrichment 87:96 enrichment 87:96 enrichment 87:96 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 52 theme glycopeptides 140:152 arg1 analysis 121:128 mass spectrometric analysis 102:128 mass spectrometric analysis 102:128 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 53 from analysis 121:128 arg1 liver 165:169 mouse liver 159:169 mouse liver 159:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 7 54 from proteins 1094:1101 arg1 data 1053:1056 comparative brain/liver glycosylation data 1015:1056 comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins 1015:1150 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 3 55 theme secreted 552:559 arg1 structures 489:498 The N-glycan structures 476:498 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins 476:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 2 56 theme sialic 275:280 arg1 variants 287:294 different sialic acid variants 265:294 different sialic acid variants 265:294 Incorporation of different sialic acid variants in both N- and O-linked glycans was observed, and the importance of using both collisional activation and electron transfer dissociation for glycopeptide analysis was illustrated. 25995273 3 57 theme lysosomal 513:521 arg1 structures 489:498 The N-glycan structures 476:498 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins 476:586 The N-glycan structures of predicted lysosomal, endoplasmic reticulum (ER), secreted and transmembrane proteins were compared. 25995273 4 58 theme protein 625:631 arg1 N-glycosylation 633:647 protein N-glycosylation 625:647 protein N-glycosylation 625:647 The data suggest that protein N-glycosylation differs depending on cellular location. 25995273 1 59 theme ∼ 232:232 arg1 proteins 238:245 ∼ 130 proteins 232:245 ∼ 130 proteins 232:245 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 0 60 dep Glycosylation 16:28 arg1 Level 50:54 the Glycopeptide Level 33:54 the Glycopeptide Level 33:54 Tissue-Specific Glycosylation at the Glycopeptide Level. 25995273 1 61 theme O-glycosylation 207:221 arg1 data 223:226 site-specific N- and O-glycosylation data 186:226 site-specific N- and O-glycosylation data for ∼ 130 proteins 186:245 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 1 62 theme mouse 159:163 arg1 liver 165:169 mouse liver 159:169 mouse liver 159:169 This manuscript describes the enrichment and mass spectrometric analysis of intact glycopeptides from mouse liver, which yielded site-specific N- and O-glycosylation data for ∼ 130 proteins. 25995273 7 63 from data 1053:1056 arg1 sites 1080:1084 50 N-glycosylation sites 1061:1084 50 N-glycosylation sites 1061:1084 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 25995273 4 64 theme cellular 670:677 arg1 location 679:686 cellular location 670:686 cellular location 670:686 The data suggest that protein N-glycosylation differs depending on cellular location. 25995273 7 65 theme glycosylation 1039:1051 arg1 data 1053:1056 comparative brain/liver glycosylation data 1015:1056 comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins 1015:1150 We present comparative brain/liver glycosylation data on 50 N-glycosylation sites from 34 proteins and 13 O-glycosylation sites from seven proteins. 28860626 6 0 gly fucosylation 1314:1325 arg1 N-glycans 1364:1372 N-glycans 1364:1372 N-glycans 1364:1372 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 0 gly fucosylation 1314:1325 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 0 gly fucosylation 1314:1325 arg1 sites 1434:1438 the specific glycosylation sites 1407:1438 the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1407:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 1 theme glycosylation 1420:1432 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 1 theme glycosylation 1420:1432 arg1 sites 1434:1438 the specific glycosylation sites 1407:1438 the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1407:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 2 theme reassortants 1596:1607 arg1 abundance 1563:1571 the protein abundance 1551:1571 the protein abundance of the influenza virus reassortants 1551:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 2 3 dep vaccines 583:590 arg1 NYMC-X181A 609:618 NYMC-X181A 609:618 NYMC-X181A 609:618 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 3 dep vaccines 583:590 arg1 NIBRG-121xp 593:603 NIBRG-121xp 593:603 NIBRG-121xp 593:603 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 3 dep vaccines 583:590 arg1 vaccines 583:590 two high-yield candidate reassortant vaccines 546:590 two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) 546:619 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 4 theme various 634:640 arg1 digestions 652:661 various enzymatic digestions 634:661 various enzymatic digestions 634:661 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 5 5 dep structure 1143:1151 arg1 location 1169:1176 location 1169:1176 location 1169:1176 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 1 6 from deletion 323:330 arg1 neuraminidase 400:412 neuraminidase 400:412 neuraminidase 400:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 6 from deletion 323:330 arg1 hemagglutinin 382:394 the influenza proteins hemagglutinin 359:394 the influenza proteins hemagglutinin 359:394 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 2 7 theme liquid 685:690 arg1 chromatography 692:705 high performance liquid chromatography 668:705 high performance liquid chromatography 668:705 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 8 theme protein 788:794 arg1 sequences 796:804 the full-length protein sequences 772:804 the full-length protein sequences 772:804 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 1 9 from addition 311:318 arg1 neuraminidase 400:412 neuraminidase 400:412 neuraminidase 400:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 9 from addition 311:318 arg1 hemagglutinin 382:394 the influenza proteins hemagglutinin 359:394 the influenza proteins hemagglutinin 359:394 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 2 10 theme high 668:671 arg1 chromatography 692:705 high performance liquid chromatography 668:705 high performance liquid chromatography 668:705 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 11 theme N-glycosylation 819:833 arg1 hemagglutinin 844:856 hemagglutinin 844:856 hemagglutinin 844:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 11 theme N-glycosylation 819:833 arg1 sites 835:839 9 N-glycosylation sites 817:839 9 N-glycosylation sites of hemagglutinin 817:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 6 12 theme proteins 1461:1468 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 12 theme proteins 1461:1468 arg1 sites 1434:1438 the specific glycosylation sites 1407:1438 the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1407:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 5 13 theme N-glycans 1187:1195 arg1 topological 1157:1167 topological 1157:1167 topological 1157:1167 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 5 13 theme N-glycans 1187:1195 arg1 structure 1143:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 1 14 theme high-yielding 219:231 arg1 vaccines 233:240 high-yielding vaccines 219:240 high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 219:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 6 15 theme bisecting 1331:1339 arg1 N-acetylglucosamine 1341:1359 bisecting N-acetylglucosamine 1331:1359 bisecting N-acetylglucosamine 1331:1359 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 3 16 theme abundance 928:936 arg1 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 2 17 theme glycosylation 528:540 arg1 topology 516:523 topology 516:523 topology 516:523 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 17 theme glycosylation 528:540 arg1 composition 475:485 glycan composition 468:485 glycan composition 468:485 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 17 theme glycosylation 528:540 arg1 distribution 499:510 structural distribution 488:510 structural distribution 488:510 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 18 gly N-glycosylation 873:887 arg2 6 871:871 6 871:871 6 871:871 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 18 gly N-glycosylation 873:887 arg1 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 18 gly N-glycosylation 873:887 arg2 sites 889:893 6 N-glycosylation sites 871:893 6 N-glycosylation sites 871:893 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 18 gly N-glycosylation 873:887 arg2 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 19 theme N-glycosylation 873:887 arg1 sites 889:893 6 N-glycosylation sites 871:893 6 N-glycosylation sites 871:893 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 19 theme N-glycosylation 873:887 arg1 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 6 20 theme protein 1518:1524 arg1 structure 1526:1534 the protein structure 1514:1534 the protein structure 1514:1534 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 5 21 theme spatial 1226:1232 arg1 structure 1242:1250 the spatial protein structure 1222:1250 the spatial protein structure 1222:1250 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 1 22 theme pandemic 154:161 arg1 influenza 163:171 a pandemic influenza H1N1 152:176 a pandemic influenza H1N1 152:176 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 4 23 theme ~300 1036:1039 arg1 glycopeptides 1041:1053 ~300 glycopeptides 1036:1053 ~300 glycopeptides 1036:1053 A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. 28860626 2 24 theme multiple-stage 711:724 arg1 spectrometry 731:742 multiple-stage mass spectrometry 711:742 multiple-stage mass spectrometry 711:742 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 25 theme glycan 903:908 arg1 structures 910:919 the glycan structures 899:919 the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans 899:1022 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 0 26 theme expressed 101:109 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 1 27 theme influenza 363:371 arg1 hemagglutinin 382:394 the influenza proteins hemagglutinin 359:394 the influenza proteins hemagglutinin 359:394 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 0 28 theme candidate 116:124 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 1 29 from outbreak 140:147 arg1 2009 181:184 2009 181:184 2009 181:184 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 2 30 theme glycan 468:473 arg1 composition 475:485 glycan composition 468:485 glycan composition 468:485 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 31 gly N-glycosylation 819:833 arg2 9 817:817 9 817:817 9 817:817 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 31 gly N-glycosylation 819:833 arg1 hemagglutinin 844:856 hemagglutinin 844:856 hemagglutinin 844:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 31 gly N-glycosylation 819:833 arg2 hemagglutinin 844:856 hemagglutinin 844:856 hemagglutinin 844:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 31 gly N-glycosylation 819:833 arg2 sites 835:839 9 N-glycosylation sites 817:839 9 N-glycosylation sites of hemagglutinin 817:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 1 32 dep influenza 163:171 arg1 H1N1 173:176 H1N1 173:176 H1N1 173:176 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 0 33 theme Topological 0:10 arg1 N-glycosylation 12:26 Topological N-glycosylation 0:26 Topological N-glycosylation 0:26 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 4 34 theme tandem 1095:1100 arg1 spectrometry 1107:1118 tandem mass spectrometry 1095:1118 tandem mass spectrometry 1095:1118 A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. 28860626 0 35 theme N-glycan 46:53 arg1 sulfation 55:63 site-specific N-glycan sulfation 32:63 site-specific N-glycan sulfation 32:63 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 0 36 gly N-glycosylation 12:26 arg1 proteins 78:85 influenza proteins 68:85 influenza proteins 68:85 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 0 36 gly N-glycosylation 12:26 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 0 37 theme proteins 78:85 arg1 sulfation 55:63 site-specific N-glycan sulfation 32:63 site-specific N-glycan sulfation 32:63 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 0 37 theme proteins 78:85 arg1 N-glycosylation 12:26 Topological N-glycosylation 0:26 Topological N-glycosylation 0:26 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 1 38 from site 351:354 arg1 neuraminidase 400:412 neuraminidase 400:412 neuraminidase 400:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 38 from site 351:354 arg1 hemagglutinin 382:394 the influenza proteins hemagglutinin 359:394 the influenza proteins hemagglutinin 359:394 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 5 39 theme ligand 1272:1277 arg1 binding 1279:1285 the residential ligand binding 1256:1285 the residential ligand binding 1256:1285 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 1 40 theme glycosylation 337:349 arg1 site 351:354 a glycosylation site 335:354 a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 335:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 5 41 theme N-glycan 1134:1141 arg1 structure 1143:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 3 42 theme data 755:758 arg1 analyses 760:767 Proteomic data analyses 745:767 Proteomic data analyses of the full-length protein sequences 745:804 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 1 43 theme vaccines 233:240 arg1 generation 205:214 the rapid generation 195:214 the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 195:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 3 44 theme full-length 776:786 arg1 sequences 796:804 the full-length protein sequences 772:804 the full-length protein sequences 772:804 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 5 45 theme protein 1234:1240 arg1 structure 1242:1250 the spatial protein structure 1222:1250 the spatial protein structure 1222:1250 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 2 46 theme enzymatic 642:650 arg1 digestions 652:661 various enzymatic digestions 634:661 various enzymatic digestions 634:661 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 6 47 theme specific 1411:1418 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 47 theme specific 1411:1418 arg1 sites 1434:1438 the specific glycosylation sites 1407:1438 the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1407:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 48 theme virus 1590:1594 arg1 reassortants 1596:1607 the influenza virus reassortants 1576:1607 the influenza virus reassortants 1576:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 1 49 gly glycosylation 337:349 arg2 site 351:354 a glycosylation site 335:354 a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 335:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 2 50 theme performance 673:683 arg1 chromatography 692:705 high performance liquid chromatography 668:705 high performance liquid chromatography 668:705 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 51 theme sequences 796:804 arg1 analyses 760:767 Proteomic data analyses 745:767 Proteomic data analyses of the full-length protein sequences 745:804 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 52 theme hemagglutinin 844:856 arg1 hemagglutinin 844:856 hemagglutinin 844:856 hemagglutinin 844:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 52 theme hemagglutinin 844:856 arg1 sites 835:839 9 N-glycosylation sites 817:839 9 N-glycosylation sites of hemagglutinin 817:856 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 1 53 theme rapid 199:203 arg1 generation 205:214 the rapid generation 195:214 the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 195:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 2 54 theme reassortant 571:581 arg1 NYMC-X181A 609:618 NYMC-X181A 609:618 NYMC-X181A 609:618 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 54 theme reassortant 571:581 arg1 NIBRG-121xp 593:603 NIBRG-121xp 593:603 NIBRG-121xp 593:603 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 54 theme reassortant 571:581 arg1 vaccines 583:590 two high-yield candidate reassortant vaccines 546:590 two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) 546:619 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 6 55 theme N-glycans 1364:1372 arg1 fucosylation 1314:1325 fucosylation 1314:1325 fucosylation 1314:1325 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 55 theme N-glycans 1364:1372 arg1 N-acetylglucosamine 1341:1359 bisecting N-acetylglucosamine 1331:1359 bisecting N-acetylglucosamine 1331:1359 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 55 theme N-glycans 1364:1372 arg1 sulfation 1303:1311 sulfation 1303:1311 sulfation 1303:1311 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 2 56 theme candidate 561:569 arg1 NYMC-X181A 609:618 NYMC-X181A 609:618 NYMC-X181A 609:618 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 56 theme candidate 561:569 arg1 NIBRG-121xp 593:603 NIBRG-121xp 593:603 NIBRG-121xp 593:603 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 56 theme candidate 561:569 arg1 vaccines 583:590 two high-yield candidate reassortant vaccines 546:590 two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) 546:619 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 6 57 theme influenza 1451:1459 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 3 58 used occupied 964:971 arg2 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 6 59 gly glycosylation 1420:1432 arg2 sites 1434:1438 the specific glycosylation sites 1407:1438 the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1407:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 59 gly glycosylation 1420:1432 arg2 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 6 59 gly glycosylation 1420:1432 arg1 proteins 1461:1468 the two influenza proteins 1443:1468 the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants 1443:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 3 60 theme neuraminidase 938:950 arg1 structures 910:919 the glycan structures 899:919 the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans 899:1022 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 60 theme neuraminidase 938:950 arg1 sites 889:893 6 N-glycosylation sites 871:893 6 N-glycosylation sites 871:893 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 3 60 theme neuraminidase 938:950 arg1 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 1 61 theme site 351:354 arg1 deletion 323:330 deletion 323:330 deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 323:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 61 theme site 351:354 arg1 addition 311:318 addition 311:318 addition 311:318 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 62 theme influenza 163:171 arg1 outbreak 140:147 The outbreak 136:147 The outbreak of a pandemic influenza H1N1 in 2009 136:184 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 0 63 from N-glycosylation 12:26 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 2 64 theme mass 726:729 arg1 spectrometry 731:742 multiple-stage mass spectrometry 711:742 multiple-stage mass spectrometry 711:742 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 3 65 theme low 924:926 arg1 neuraminidase 938:950 low abundance neuraminidase 924:950 low abundance neuraminidase 924:950 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 0 66 theme H1N1 111:114 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 2 67 theme high-yield 550:559 arg1 NYMC-X181A 609:618 NYMC-X181A 609:618 NYMC-X181A 609:618 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 67 theme high-yield 550:559 arg1 NIBRG-121xp 593:603 NIBRG-121xp 593:603 NIBRG-121xp 593:603 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 2 67 theme high-yield 550:559 arg1 vaccines 583:590 two high-yield candidate reassortant vaccines 546:590 two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) 546:619 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 1 68 from neuraminidase 400:412 arg1 deletion 323:330 deletion 323:330 deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 323:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 68 from neuraminidase 400:412 arg1 addition 311:318 addition 311:318 addition 311:318 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 69 theme proteins 373:380 arg1 hemagglutinin 382:394 the influenza proteins hemagglutinin 359:394 the influenza proteins hemagglutinin 359:394 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 3 70 theme complex-type 1001:1012 arg1 N-glycans 1014:1022 complex-type N-glycans 1001:1022 complex-type N-glycans 1001:1022 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 4 71 theme glycopeptides 1041:1053 arg1 total 1027:1031 A total 1025:1031 A total of ~300 glycopeptides 1025:1053 A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. 28860626 0 72 from sulfation 55:63 arg1 vaccines 126:133 the highly expressed H1N1 candidate vaccines 90:133 the highly expressed H1N1 candidate vaccines 90:133 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 6 73 theme crucial 1487:1493 arg1 role 1495:1498 a crucial role 1485:1498 a crucial role 1485:1498 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 1 74 from hemagglutinin 382:394 arg1 deletion 323:330 deletion 323:330 deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase 323:412 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 1 74 from hemagglutinin 382:394 arg1 addition 311:318 addition 311:318 addition 311:318 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 28860626 0 75 theme site-specific 32:44 arg1 sulfation 55:63 site-specific N-glycan sulfation 32:63 site-specific N-glycan sulfation 32:63 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 4 76 gly glycopeptides 1041:1053 arg2 glycopeptides 1041:1053 ~300 glycopeptides 1036:1053 ~300 glycopeptides 1036:1053 A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. 28860626 6 77 theme protein 1555:1561 arg1 abundance 1563:1571 the protein abundance 1551:1571 the protein abundance of the influenza virus reassortants 1551:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 2 78 theme structural 488:497 arg1 distribution 499:510 structural distribution 488:510 structural distribution 488:510 In this report, we have systematically evaluated the glycan composition, structural distribution and topology of glycosylation for two high-yield candidate reassortant vaccines (NIBRG-121xp and NYMC-X181A) by combining various enzymatic digestions with high performance liquid chromatography and multiple-stage mass spectrometry. 28860626 0 79 theme influenza 68:76 arg1 proteins 78:85 influenza proteins 68:85 influenza proteins 68:85 Topological N-glycosylation and site-specific N-glycan sulfation of influenza proteins in the highly expressed H1N1 candidate vaccines. 28860626 5 80 theme residential 1260:1270 arg1 binding 1279:1285 the residential ligand binding 1256:1285 the residential ligand binding 1256:1285 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 5 81 theme specific 1125:1132 arg1 structure 1143:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure 1121:1151 The specific N-glycan structure and topological location of these N-glycans are highly correlated to the spatial protein structure and the residential ligand binding. 28860626 4 82 theme mass 1102:1105 arg1 spectrometry 1107:1118 tandem mass spectrometry 1095:1118 tandem mass spectrometry 1095:1118 A total of ~300 glycopeptides were analyzed and manually validated by tandem mass spectrometry. 28860626 3 83 theme Proteomic 745:753 arg1 analyses 760:767 Proteomic data analyses 745:767 Proteomic data analyses of the full-length protein sequences 745:804 Proteomic data analyses of the full-length protein sequences determined 9 N-glycosylation sites of hemagglutinin, and defined 6 N-glycosylation sites and the glycan structures of low abundance neuraminidase, which were occupied by high-mannose, hybrid and complex-type N-glycans. 28860626 6 84 theme influenza 1580:1588 arg1 reassortants 1596:1607 the influenza virus reassortants 1576:1607 the influenza virus reassortants 1576:1607 Interestingly, sulfation, fucosylation and bisecting N-acetylglucosamine of N-glycans were also reliably identified at the specific glycosylation sites of the two influenza proteins that may serve a crucial role in regulating the protein structure and increasing the protein abundance of the influenza virus reassortants. 28860626 1 85 theme A/California/7/2009 254:272 arg1 virus 274:278 the A/California/7/2009 virus 250:278 the A/California/7/2009 virus 250:278 The outbreak of a pandemic influenza H1N1 in 2009 required the rapid generation of high-yielding vaccines against the A/California/7/2009 virus, which were achieved by either addition or deletion of a glycosylation site in the influenza proteins hemagglutinin and neuraminidase. 25029371 3 0 gly glycopeptide 601:612 arg2 glycopeptide 601:612 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 7 1 theme STT3B 1515:1519 arg1 isoforms 1521:1528 the OST STT3A and STT3B isoforms 1497:1528 the OST STT3A and STT3B isoforms 1497:1528 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 6 2 theme barring 1185:1191 arg1 sites 1178:1182 NXT consensus sites 1164:1182 NXT consensus sites (barring NWT) 1164:1196 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 2 theme barring 1185:1191 arg1 NWT 1193:1195 barring NWT 1185:1195 barring NWT 1185:1195 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 8 3 theme OST 1932:1934 arg1 isoforms 1936:1943 the mammalian OST isoforms 1918:1943 the mammalian OST isoforms 1918:1943 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 2 4 theme nascent 287:293 arg1 polypeptides 295:306 nascent polypeptides 287:306 nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), 287:343 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 7 5 theme STT3A 1505:1509 arg1 isoforms 1521:1528 the OST STT3A and STT3B isoforms 1497:1528 the OST STT3A and STT3B isoforms 1497:1528 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 8 6 theme homologous 1672:1681 arg1 structure 1683:1691 an OST eubacterial homologous structure 1653:1691 an OST eubacterial homologous structure 1653:1691 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 3 7 theme site 480:483 arg1 X 485:485 the consensus site X and hydroxyl residue 466:506 X 485:485 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 1 8 theme endoplasmic 175:185 arg1 reticulum 187:195 the endoplasmic reticulum 171:195 the endoplasmic reticulum (ER) 171:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 1 8 theme endoplasmic 175:185 arg1 ER 198:199 ER 198:199 ER 198:199 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 4 9 theme charged 830:836 arg1 residues 845:852 large hydrophobic and negatively charged middle residues 797:852 large hydrophobic and negatively charged middle residues 797:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 7 10 theme post-translational 1433:1450 arg1 N-glycosylation 1452:1466 post-translational N-glycosylation 1433:1466 post-translational N-glycosylation 1433:1466 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 4 11 theme rapid 630:634 arg1 experiments 660:670 rapid radioactive pulse-chase experiments 630:670 rapid radioactive pulse-chase experiments 630:670 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 8 12 theme binding 1773:1779 arg1 site 1781:1784 the peptide binding site 1761:1784 the peptide binding site 1761:1784 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 1 13 gly glycosylation 111:123 arg1 protein 152:158 efficient protein folding 142:166 efficient protein folding in the endoplasmic reticulum (ER) 142:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 1 13 gly glycosylation 111:123 arg1 anterograde 206:216 anterograde trafficking 206:228 anterograde trafficking through the secretory pathway 206:258 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 3 14 theme residue 500:506 arg1 effect 456:461 the effect 452:461 the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 452:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 6 15 mod modified 1215:1222 arg3 machinery 1247:1255 the cotranslational machinery 1227:1255 the cotranslational machinery 1227:1255 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 15 mod modified 1215:1222 arg1 sites 1178:1182 NXT consensus sites 1164:1182 NXT consensus sites (barring NWT) 1164:1196 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 15 mod modified 1215:1222 arg1 NWT 1193:1195 barring NWT 1185:1195 barring NWT 1185:1195 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 16 theme cotranslational 1231:1245 arg1 machinery 1247:1255 the cotranslational machinery 1227:1255 the cotranslational machinery 1227:1255 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 17 theme protein 1324:1330 arg1 translation 1332:1342 protein translation 1324:1342 protein translation 1324:1342 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 5 18 theme sites 1080:1084 arg1 hypoglycosylation 1055:1071 hypoglycosylation 1055:1071 hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains 1055:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 8 19 theme hydrophobic 1800:1810 arg1 residues 1835:1842 large hydrophobic and negatively charged residues 1794:1842 large hydrophobic and negatively charged residues 1794:1842 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 1 20 theme anterograde 206:216 arg1 trafficking 218:228 anterograde trafficking 206:228 anterograde trafficking through the secretory pathway 206:258 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 7 21 theme N-glycosylation 1390:1404 arg1 efficiency 1406:1415 cotranslational N-glycosylation efficiency 1374:1415 cotranslational N-glycosylation efficiency 1374:1415 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 4 22 theme co-translational 683:698 arg1 events 739:744 co-translational (STT3A) and post-translational (STT3B) events 683:744 co-translational (STT3A) and post-translational (STT3B) events 683:744 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 6 23 theme consensus 1293:1301 arg1 sites 1303:1307 consensus sites 1293:1307 consensus sites skipped during protein translation 1293:1342 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 5 24 theme Post-translational 914:931 arg1 modification 933:944 Post-translational modification 914:944 Post-translational modification of the cotranslationally skipped sites by STT3B 914:992 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 7 25 theme strong 1347:1352 arg1 correlation 1354:1364 A strong correlation 1345:1364 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation 1345:1466 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 4 26 theme pulse-chase 648:658 arg1 experiments 660:670 rapid radioactive pulse-chase experiments 630:670 rapid radioactive pulse-chase experiments 630:670 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 5 27 theme charged 1130:1136 arg1 chains 1143:1148 large hydrophobic and negatively charged side chains 1097:1148 large hydrophobic and negatively charged side chains 1097:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 3 28 theme N-glycosylation 559:573 arg1 distributions 515:527 the distributions 511:527 the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 511:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 1 29 theme protein 152:158 arg1 folding 160:166 efficient protein folding 142:166 efficient protein folding in the endoplasmic reticulum (ER) 142:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 4 30 contain containing 786:795 arg1 sites 780:784 NXS consensus sites 766:784 NXS consensus sites containing large hydrophobic and negatively charged middle residues 766:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 4 30 contain containing 786:795 arg2 residues 845:852 large hydrophobic and negatively charged middle residues 797:852 large hydrophobic and negatively charged middle residues 797:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 1 31 from folding 160:166 arg1 reticulum 187:195 the endoplasmic reticulum 171:195 the endoplasmic reticulum (ER) 171:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 1 31 from folding 160:166 arg1 ER 198:199 ER 198:199 ER 198:199 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 5 32 theme large 1097:1101 arg1 chains 1143:1148 large hydrophobic and negatively charged side chains 1097:1148 large hydrophobic and negatively charged side chains 1097:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 5 33 theme sites 979:983 arg1 modification 933:944 Post-translational modification 914:944 Post-translational modification of the cotranslationally skipped sites by STT3B 914:992 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 7 34 theme site 1594:1597 arg1 residues 1599:1606 the hydroxyl and middle X consensus site residues 1558:1606 residues 1599:1606 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 2 35 theme enzymatic 359:367 arg1 isoforms 369:376 two enzymatic isoforms 355:376 two enzymatic isoforms of the oligosaccharyltransferase (OST) 355:415 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 8 36 theme enzymatic 1893:1901 arg1 mechanisms 1903:1912 conserved enzymatic mechanisms 1883:1912 conserved enzymatic mechanisms for the mammalian OST isoforms 1883:1943 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 5 37 contain containing 1086:1095 arg2 chains 1143:1148 large hydrophobic and negatively charged side chains 1097:1148 large hydrophobic and negatively charged side chains 1097:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 5 37 contain containing 1086:1095 arg1 sites 1080:1084 NXS sites 1076:1084 NXS sites containing large hydrophobic and negatively charged side chains 1076:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 2 38 theme oligosaccharyltransferase 385:409 arg1 isoforms 369:376 two enzymatic isoforms 355:376 two enzymatic isoforms of the oligosaccharyltransferase (OST) 355:415 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 0 39 theme X 11:11 arg1 residue 13:19 The middle X residue 0:19 The middle X residue 0:19 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 1 40 theme -linked 103:109 arg1 glycosylation 111:123 Asparagine (N)-linked glycosylation 89:123 Asparagine (N)-linked glycosylation 89:123 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 3 41 theme glycopeptide 601:612 arg1 scaffold 614:621 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 5 42 theme middle 1024:1029 arg1 residue 1033:1039 the middle X residue 1020:1039 the middle X residue 1020:1039 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 3 43 theme I 585:585 arg1 scaffold 614:621 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 4 44 theme large 797:801 arg1 residues 845:852 large hydrophobic and negatively charged middle residues 797:852 large hydrophobic and negatively charged middle residues 797:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 2 45 theme ≠ 339:339 arg1 N-X-T/S 328:334 N-X-T/S 328:334 N-X-T/S (X ≠ P) 328:342 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 45 theme ≠ 339:339 arg1 P 341:341 X ≠ P 337:341 X ≠ P 337:341 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 0 46 theme N-glycosylation 48:62 arg1 site 74:77 cotranslational N-glycosylation consensus site 32:77 cotranslational N-glycosylation consensus site skipping 32:86 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 8 47 theme middle 1630:1635 arg1 residues 1639:1646 various middle X residues 1622:1646 various middle X residues 1622:1646 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 4 48 theme NXS 766:768 arg1 sites 780:784 NXS consensus sites 766:784 NXS consensus sites containing large hydrophobic and negatively charged middle residues 766:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 8 49 theme mammalian 1922:1930 arg1 isoforms 1936:1943 the mammalian OST isoforms 1918:1943 the mammalian OST isoforms 1918:1943 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 8 50 theme consensus 1723:1731 arg1 residues 1740:1747 small and polar consensus site X residues 1707:1747 small and polar consensus site X residues 1707:1747 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 4 51 theme middle 838:843 arg1 residues 845:852 large hydrophobic and negatively charged middle residues 797:852 large hydrophobic and negatively charged middle residues 797:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 5 52 gly hypoglycosylation 1055:1071 arg1 sites 1080:1084 NXS sites 1076:1084 NXS sites containing large hydrophobic and negatively charged side chains 1076:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 8 53 theme eubacterial 1660:1670 arg1 structure 1683:1691 an OST eubacterial homologous structure 1653:1691 an OST eubacterial homologous structure 1653:1691 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 3 54 from effect 456:461 arg1 distributions 515:527 the distributions 511:527 the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 511:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 3 55 theme X 485:485 arg1 effect 456:461 the effect 452:461 the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 452:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 8 56 theme X 1738:1738 arg1 residues 1740:1747 small and polar consensus site X residues 1707:1747 small and polar consensus site X residues 1707:1747 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 6 57 theme NXT 1164:1166 arg1 sites 1178:1182 NXT consensus sites 1164:1182 NXT consensus sites (barring NWT) 1164:1196 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 57 theme NXT 1164:1166 arg1 NWT 1193:1195 barring NWT 1185:1195 barring NWT 1185:1195 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 7 58 theme OST 1501:1503 arg1 STT3A 1505:1509 OST STT3A 1501:1509 OST STT3A 1501:1509 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 3 59 theme consensus 470:478 arg1 X 485:485 the consensus site X and hydroxyl residue 466:506 X 485:485 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 3 60 gly N-glycosylation 559:573 arg1 scaffold 614:621 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 4 61 theme hydrophobic 803:813 arg1 residues 845:852 large hydrophobic and negatively charged middle residues 797:852 large hydrophobic and negatively charged middle residues 797:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 6 62 theme consensus 1168:1176 arg1 sites 1178:1182 NXT consensus sites 1164:1182 NXT consensus sites (barring NWT) 1164:1196 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 6 62 theme consensus 1168:1176 arg1 NWT 1193:1195 barring NWT 1185:1195 barring NWT 1185:1195 In contrast, NXT consensus sites (barring NWT) were efficiently modified by the cotranslational machinery, reducing STT3B's role in modifying consensus sites skipped during protein translation. 25029371 7 63 theme N-glycosylation 1452:1466 arg1 efficiency 1406:1415 cotranslational N-glycosylation efficiency 1374:1415 cotranslational N-glycosylation efficiency 1374:1415 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 7 63 theme N-glycosylation 1452:1466 arg1 rate 1425:1428 the rate 1421:1428 the rate of post-translational N-glycosylation 1421:1466 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 2 64 theme consensus 311:319 arg1 N-X-T/S 328:334 N-X-T/S 328:334 N-X-T/S (X ≠ P) 328:342 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 64 theme consensus 311:319 arg1 sites 321:325 consensus sites 311:325 consensus sites 311:325 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 8 65 theme peptide 1765:1771 arg1 site 1781:1784 the peptide binding site 1761:1784 the peptide binding site 1761:1784 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 4 66 theme protein 893:899 arg1 translation 901:911 protein translation 893:911 protein translation 893:911 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 5 67 theme NXS 1076:1078 arg1 sites 1080:1084 NXS sites 1076:1084 NXS sites containing large hydrophobic and negatively charged side chains 1076:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 3 68 theme co- 532:534 arg1 N-glycosylation 559:573 co- and post-translational N-glycosylation 532:573 co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 532:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 8 69 theme large 1794:1798 arg1 residues 1835:1842 large hydrophobic and negatively charged residues 1794:1842 large hydrophobic and negatively charged residues 1794:1842 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 2 70 attach attached 275:282 arg3 STT3A 418:422 STT3A 418:422 STT3A 418:422 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 70 attach attached 275:282 arg2 N-Glycans 261:269 N-Glycans 261:269 N-Glycans 261:269 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 70 attach attached 275:282 arg3 isoforms 369:376 two enzymatic isoforms 355:376 two enzymatic isoforms of the oligosaccharyltransferase (OST) 355:415 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 70 attach attached 275:282 arg1 polypeptides 295:306 nascent polypeptides 287:306 nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), 287:343 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 70 attach attached 275:282 arg3 one 348:350 one 348:350 one 348:350 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 70 attach attached 275:282 arg3 STT3B 427:431 STT3B 427:431 STT3B 427:431 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 1 71 from trafficking 218:228 arg1 reticulum 187:195 the endoplasmic reticulum 171:195 the endoplasmic reticulum (ER) 171:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 1 71 from trafficking 218:228 arg1 ER 198:199 ER 198:199 ER 198:199 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 3 72 theme hydroxyl 491:498 arg1 residue 500:506 the consensus site X and hydroxyl residue 466:506 residue 500:506 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 7 73 theme cotranslational 1374:1388 arg1 N-glycosylation 1390:1404 cotranslational N-glycosylation 1374:1404 cotranslational N-glycosylation efficiency 1374:1415 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 3 74 theme post-translational 540:557 arg1 N-glycosylation 559:573 co- and post-translational N-glycosylation 532:573 co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 532:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 8 75 theme charged 1827:1833 arg1 residues 1835:1842 large hydrophobic and negatively charged residues 1794:1842 large hydrophobic and negatively charged residues 1794:1842 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 1 76 theme efficient 142:150 arg1 folding 160:166 efficient protein folding 142:166 efficient protein folding in the endoplasmic reticulum (ER) 142:200 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 4 77 theme radioactive 636:646 arg1 experiments 660:670 rapid radioactive pulse-chase experiments 630:670 rapid radioactive pulse-chase experiments 630:670 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 5 78 theme side 1138:1141 arg1 chains 1143:1148 large hydrophobic and negatively charged side chains 1097:1148 large hydrophobic and negatively charged side chains 1097:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 5 79 theme skipped 971:977 arg1 sites 979:983 the cotranslationally skipped sites 949:983 the cotranslationally skipped sites 949:983 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 2 80 from sites 321:325 arg1 polypeptides 295:306 nascent polypeptides 287:306 nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), 287:343 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 7 81 theme consensus 1584:1592 arg1 residues 1599:1606 the hydroxyl and middle X consensus site residues 1558:1606 residues 1599:1606 A strong correlation between cotranslational N-glycosylation efficiency and the rate of post-translational N-glycosylation was determined, showing that the OST STT3A and STT3B isoforms are similarly influenced by the hydroxyl and middle X consensus site residues. 25029371 5 82 theme hydrophobic 1103:1113 arg1 chains 1143:1148 large hydrophobic and negatively charged side chains 1097:1148 large hydrophobic and negatively charged side chains 1097:1148 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 0 83 theme middle 4:9 arg1 residue 13:19 The middle X residue 0:19 The middle X residue 0:19 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 8 84 theme conserved 1883:1891 arg1 mechanisms 1903:1912 conserved enzymatic mechanisms 1883:1912 conserved enzymatic mechanisms for the mammalian OST isoforms 1883:1943 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 4 85 dep co-translational 683:698 arg1 STT3B 732:736 STT3B 732:736 STT3B 732:736 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 4 85 dep co-translational 683:698 arg1 STT3A 701:705 STT3A 701:705 STT3A 701:705 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 3 86 theme transmembrane 587:599 arg1 scaffold 614:621 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 4 87 theme post-translational 712:729 arg1 events 739:744 co-translational (STT3A) and post-translational (STT3B) events 683:744 co-translational (STT3A) and post-translational (STT3B) events 683:744 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 0 88 theme cotranslational 32:46 arg1 site 74:77 cotranslational N-glycosylation consensus site 32:77 cotranslational N-glycosylation consensus site skipping 32:86 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 8 89 theme various 1622:1628 arg1 residues 1639:1646 various middle X residues 1622:1646 various middle X residues 1622:1646 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 3 90 theme type 580:583 arg1 scaffold 614:621 a type I transmembrane glycopeptide scaffold 578:621 a type I transmembrane glycopeptide scaffold 578:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 4 91 theme consensus 770:778 arg1 sites 780:784 NXS consensus sites 766:784 NXS consensus sites containing large hydrophobic and negatively charged middle residues 766:852 Using rapid radioactive pulse-chase experiments to resolve co-translational (STT3A) and post-translational (STT3B) events, we determined that NXS consensus sites containing large hydrophobic and negatively charged middle residues are frequently skipped by STT3A during protein translation. 25029371 2 92 theme isoforms 369:376 arg1 one 348:350 one 348:350 one 348:350 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 92 theme isoforms 369:376 arg1 STT3B 427:431 STT3B 427:431 STT3B 427:431 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 92 theme isoforms 369:376 arg1 STT3A 418:422 STT3A 418:422 STT3A 418:422 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 92 theme isoforms 369:376 arg1 isoforms 369:376 two enzymatic isoforms 355:376 two enzymatic isoforms of the oligosaccharyltransferase (OST) 355:415 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 0 93 theme consensus 64:72 arg1 site 74:77 cotranslational N-glycosylation consensus site 32:77 cotranslational N-glycosylation consensus site skipping 32:86 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 5 94 theme X 1031:1031 arg1 residue 1033:1039 the middle X residue 1020:1039 the middle X residue 1020:1039 Post-translational modification of the cotranslationally skipped sites by STT3B was similarly hindered by the middle X residue, resulting in hypoglycosylation of NXS sites containing large hydrophobic and negatively charged side chains. 25029371 8 95 theme X 1637:1637 arg1 residues 1639:1646 various middle X residues 1622:1646 various middle X residues 1622:1646 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 2 96 theme X 337:337 arg1 N-X-T/S 328:334 N-X-T/S 328:334 N-X-T/S (X ≠ P) 328:342 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 2 96 theme X 337:337 arg1 P 341:341 X ≠ P 337:341 X ≠ P 337:341 N-Glycans are attached to nascent polypeptides at consensus sites, N-X-T/S (X ≠ P), by one of two enzymatic isoforms of the oligosaccharyltransferase (OST), STT3A or STT3B. 25029371 8 97 theme small 1707:1711 arg1 residues 1740:1747 small and polar consensus site X residues 1707:1747 small and polar consensus site X residues 1707:1747 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 3 98 theme scaffold 614:621 arg1 N-glycosylation 559:573 co- and post-translational N-glycosylation 532:573 co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold 532:621 Here, we examined the effect of the consensus site X and hydroxyl residue on the distributions of co- and post-translational N-glycosylation of a type I transmembrane glycopeptide scaffold. 25029371 8 99 theme polar 1717:1721 arg1 residues 1740:1747 small and polar consensus site X residues 1707:1747 small and polar consensus site X residues 1707:1747 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 0 100 gly N-glycosylation 48:62 arg2 site 74:77 cotranslational N-glycosylation consensus site 32:77 cotranslational N-glycosylation consensus site skipping 32:86 The middle X residue influences cotranslational N-glycosylation consensus site skipping. 25029371 8 101 theme OST 1656:1658 arg1 structure 1683:1691 an OST eubacterial homologous structure 1653:1691 an OST eubacterial homologous structure 1653:1691 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 1 102 link -linked 103:109 arg1 glycosylation 111:123 Asparagine (N)-linked glycosylation 89:123 Asparagine (N)-linked glycosylation 89:123 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 25029371 8 103 theme site 1733:1736 arg1 residues 1740:1747 small and polar consensus site X residues 1707:1747 small and polar consensus site X residues 1707:1747 Substituting various middle X residues into an OST eubacterial homologous structure revealed that small and polar consensus site X residues fit well in the peptide binding site whereas large hydrophobic and negatively charged residues were harder to accommodate, indicating conserved enzymatic mechanisms for the mammalian OST isoforms. 25029371 1 104 theme secretory 242:250 arg1 pathway 252:258 the secretory pathway 238:258 the secretory pathway 238:258 Asparagine (N)-linked glycosylation is essential for efficient protein folding in the endoplasmic reticulum (ER) and anterograde trafficking through the secretory pathway. 28221766 9 0 theme detected 1548:1555 arg1 modifications 1580:1592 modifications 1580:1592 modifications on a structural model of C9 1580:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 0 theme detected 1548:1555 arg1 sites 1571:1575 all detected and validated sites 1544:1575 sites 1571:1575 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 8 1 dep two 1515:1517 arg1 to 1512:1513 to 1512:1513 to 1512:1513 Our data also reveal a binding of up to two Ca2+ ions to C9. 28221766 10 2 theme unbiased 1781:1788 arg1 analysis 1799:1806 the unbiased in-depth analysis 1777:1806 the unbiased in-depth analysis of plasma proteins 1777:1825 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 1 3 theme pathway 208:214 arg1 protein 160:166 human complement C9 protein 140:166 The human complement C9 protein (∼65 kDa) 136:176 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 3 theme pathway 208:214 arg1 member 183:188 a member 181:188 a member of the complement pathway 181:214 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 9 4 theme validated 1561:1569 arg1 modifications 1580:1592 modifications 1580:1592 modifications on a structural model of C9 1580:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 4 theme validated 1561:1569 arg1 sites 1571:1575 all detected and validated sites 1544:1575 sites 1571:1575 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 6 5 theme O-linked 1213:1220 arg1 glycans 1222:1228 O-linked glycans 1213:1228 O-linked glycans in the N-terminal region 1213:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 2 6 theme essential 229:237 arg1 role 239:242 an essential role 226:242 an essential role 226:242 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 6 7 from evidence 1201:1208 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 7 8 gly N-glycosylation 1349:1363 arg2 site 1365:1368 a third novel, albeit low abundant, N-glycosylation site 1313:1368 a third novel, albeit low abundant, N-glycosylation site on C9 1313:1374 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 6 9 link O-linked 1213:1220 arg1 glycans 1222:1228 O-linked glycans 1213:1228 O-linked glycans in the N-terminal region 1213:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 10 10 theme plasma 1811:1816 arg1 proteins 1818:1825 plasma proteins 1811:1825 plasma proteins 1811:1825 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 1 11 theme C9 157:158 arg1 protein 160:166 human complement C9 protein 140:166 The human complement C9 protein (∼65 kDa) 136:176 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 11 theme C9 157:158 arg1 kDa 173:175 ∼65 kDa 169:175 ∼65 kDa 169:175 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 11 theme C9 157:158 arg1 member 183:188 a member 181:188 a member of the complement pathway 181:214 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 4 12 theme C9 602:603 arg1 profile 591:597 The proteoform profile 576:597 The proteoform profile of C9 576:603 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 6 13 theme direct 1181:1186 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 2 14 theme membrane 251:258 arg1 MAC 276:278 MAC 276:278 MAC 276:278 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 2 14 theme membrane 251:258 arg1 complex 267:273 membrane attack complex 251:273 the membrane attack complex (MAC) 247:279 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 7 15 dep novel 1321:1325 arg1 abundant 1339:1346 abundant 1339:1346 abundant 1339:1346 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 7 15 dep novel 1321:1325 arg1 albeit 1328:1333 albeit 1328:1333 albeit 1328:1333 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 9 16 theme structural 1599:1608 arg1 model 1610:1614 a structural model 1597:1614 a structural model of C9 1597:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 6 17 theme detailed 1078:1085 arg1 specification 1087:1099 a detailed specification 1076:1099 a detailed specification 1076:1099 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 9 18 dep hints 1646:1650 arg1 Mapping 1536:1542 Mapping 1536:1542 Mapping all detected and validated sites of modifications on a structural model of C9 1536:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 5 19 theme glycosylation 1005:1017 arg1 types 993:997 three different types 977:997 three different types of C9 glycosylation 977:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 3 20 theme integrative 465:475 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 9 21 from modifications 1580:1592 arg1 model 1610:1614 a structural model 1597:1614 a structural model of C9 1597:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 6 22 theme first 1175:1179 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 10 23 theme glycosylation 1876:1888 arg1 profiles 1890:1897 aberrant glycosylation profiles 1867:1897 aberrant glycosylation profiles 1867:1897 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 5 24 theme proteolytic 789:799 arg1 digestion 801:809 proteolytic digestion 789:809 proteolytic digestion of C9 789:815 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 9 25 from sites 1571:1575 arg1 model 1610:1614 a structural model 1597:1614 a structural model of C9 1597:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 4 26 theme mass 644:647 arg1 spectrometry 649:660 high-resolution native mass spectrometry 621:660 high-resolution native mass spectrometry 621:660 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 0 27 theme C-Glycosylation 119:133 arg1 Features 95:102 Unexpected New Features 80:102 Unexpected New Features of N-, O-, and C-Glycosylation 80:133 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 5 28 theme different 983:991 arg1 types 993:997 three different types 977:997 three different types of C9 glycosylation 977:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 3 29 theme native 512:517 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 5 30 theme -tandem 847:853 arg1 MS/MS 874:878 MS/MS 874:878 MS/MS 874:878 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 30 theme -tandem 847:853 arg1 spectrometry 860:871 liquid chromatography (LC)-tandem mass spectrometry 821:871 liquid chromatography (LC)-tandem mass spectrometry (MS/MS) 821:879 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 7 31 theme glycosylation 1292:1304 arg1 sites 1306:1310 the two known glycosylation sites 1278:1310 the two known glycosylation sites 1278:1310 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 6 32 theme C9 1152:1153 arg1 proteoforms 1155:1165 15 co-occurring C9 proteoforms 1136:1165 15 co-occurring C9 proteoforms 1136:1165 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 33 theme N-terminal 1237:1246 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 5 34 theme spectrometry 860:871 arg1 measurements 881:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements 789:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures 789:926 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 0 35 theme Component 57:65 arg1 C9 67:68 the Human Serum Complement Component C9 30:68 the Human Serum Complement Component C9 30:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 5 36 theme chromatography 828:841 arg1 MS/MS 874:878 MS/MS 874:878 MS/MS 874:878 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 36 theme chromatography 828:841 arg1 spectrometry 860:871 liquid chromatography (LC)-tandem mass spectrometry 821:871 liquid chromatography (LC)-tandem mass spectrometry (MS/MS) 821:879 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 4 37 theme high-resolution 621:635 arg1 spectrometry 649:660 high-resolution native mass spectrometry 621:660 high-resolution native mass spectrometry 621:660 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 10 38 theme state 1943:1947 arg1 indicative 1906:1915 indicative 1906:1915 indicative 1906:1915 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 1 39 theme human 140:144 arg1 protein 160:166 human complement C9 protein 140:166 The human complement C9 protein (∼65 kDa) 136:176 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 39 theme human 140:144 arg1 kDa 173:175 ∼65 kDa 169:175 ∼65 kDa 169:175 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 39 theme human 140:144 arg1 member 183:188 a member 181:188 a member of the complement pathway 181:214 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 7 40 from site 1365:1368 arg1 C9 1373:1374 C9 1373:1374 C9 1373:1374 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 0 41 theme New 91:93 arg1 Features 95:102 Unexpected New Features 80:102 Unexpected New Features of N-, O-, and C-Glycosylation 80:133 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 9 42 from model 1610:1614 arg1 modifications 1580:1592 modifications 1580:1592 modifications on a structural model of C9 1580:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 42 from model 1610:1614 arg1 sites 1571:1575 all detected and validated sites 1544:1575 sites 1571:1575 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 43 theme putative 1661:1668 arg1 roles 1670:1674 their putative roles 1655:1674 their putative roles in pore formation or receptor interactions 1655:1717 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 4 44 theme mass 712:715 arg1 MS 731:732 MS 731:732 MS 731:732 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 4 44 theme mass 712:715 arg1 spectrometry 717:728 mass spectrometry 712:728 ∼50 distinct mass spectrometry (MS) signals 699:741 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 6 45 from validation 1102:1111 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 5 46 theme MS 1048:1049 arg1 data 1051:1054 the native MS data 1037:1054 the native MS data 1037:1054 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 47 theme peptide 911:917 arg1 mixtures 919:926 the resulting peptide mixtures 897:926 the resulting peptide mixtures 897:926 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 0 48 theme Proteoform 0:9 arg1 Mapping 19:25 Proteoform Profile Mapping 0:25 Proteoform Profile Mapping of the Human Serum Complement Component C9 0:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 2 49 theme pathogenic 335:344 arg1 bacteria 346:353 pathogenic bacteria 335:353 pathogenic bacteria 335:353 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 5 50 theme peptide-centric 755:769 arg1 analysis 771:778 Subsequent peptide-centric analysis 744:778 Subsequent peptide-centric analysis 744:778 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 9 51 theme receptor 1697:1704 arg1 interactions 1706:1717 receptor interactions 1697:1717 receptor interactions 1697:1717 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 6 52 from specification 1087:1099 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 5 53 theme digestion 801:809 arg1 measurements 881:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements 789:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures 789:926 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 6 54 from region 1248:1253 arg1 specification 1087:1099 a detailed specification 1076:1099 a detailed specification 1076:1099 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 54 from region 1248:1253 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 54 from region 1248:1253 arg1 validation 1102:1111 validation 1102:1111 validation 1102:1111 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 54 from region 1248:1253 arg1 quantification 1118:1131 quantification 1118:1131 quantification 1118:1131 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 7 55 theme sequence 1457:1464 arg1 N-X-S/T 1466:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 2 56 from pore 303:306 arg1 surface 324:330 the cellular surface 311:330 the cellular surface of pathogenic bacteria 311:353 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 10 57 theme applied 1724:1730 arg1 methods 1732:1738 The applied methods 1720:1738 The applied methods 1720:1738 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 0 58 theme Serum 40:44 arg1 C9 67:68 the Human Serum Complement Component C9 30:68 the Human Serum Complement Component C9 30:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 7 59 theme canonical 1431:1439 arg1 N-X-S/T 1466:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 6 60 from glycans 1222:1228 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 3 61 dep charted 365:371 arg1 using 456:460 using 456:460 using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics 456:573 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 7 62 contain possess 1419:1425 arg2 N-X-S/T 1466:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 7 62 contain possess 1419:1425 arg1 which 1391:1395 which 1391:1395 which 1391:1395 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 5 63 theme site-specific 938:950 arg1 profiles 965:972 site-specific quantitative profiles 938:972 site-specific quantitative profiles of three different types of C9 glycosylation 938:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 8 64 theme ions 1524:1527 arg1 binding 1498:1504 a binding 1496:1504 a binding of up to two Ca2+ ions to C9 1496:1533 Our data also reveal a binding of up to two Ca2+ ions to C9. 28221766 10 65 theme in-depth 1790:1797 arg1 analysis 1799:1806 the unbiased in-depth analysis 1777:1806 the unbiased in-depth analysis of plasma proteins 1777:1825 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 3 66 theme C9 420:421 arg1 microheterogeneity 398:415 the structural microheterogeneity 383:415 the structural microheterogeneity 383:415 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 1 67 theme complement 197:206 arg1 pathway 208:214 the complement pathway 193:214 the complement pathway 193:214 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 6 68 theme glycans 1222:1228 arg1 specification 1087:1099 a detailed specification 1076:1099 a detailed specification 1076:1099 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 68 theme glycans 1222:1228 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 68 theme glycans 1222:1228 arg1 validation 1102:1111 validation 1102:1111 validation 1102:1111 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 68 theme glycans 1222:1228 arg1 quantification 1118:1131 quantification 1118:1131 quantification 1118:1131 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 3 69 theme workflow 477:484 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 10 70 theme proteins 1818:1825 arg1 analysis 1799:1806 the unbiased in-depth analysis 1777:1806 the unbiased in-depth analysis of plasma proteins 1777:1825 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 3 71 theme blood 443:447 arg1 serum 449:453 human blood serum 437:453 human blood serum 437:453 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 1 72 theme complement 146:155 arg1 protein 160:166 human complement C9 protein 140:166 The human complement C9 protein (∼65 kDa) 136:176 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 72 theme complement 146:155 arg1 kDa 173:175 ∼65 kDa 169:175 ∼65 kDa 169:175 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 1 72 theme complement 146:155 arg1 member 183:188 a member 181:188 a member of the complement pathway 181:214 The human complement C9 protein (∼65 kDa) is a member of the complement pathway. 28221766 9 73 theme modifications 1580:1592 arg1 modifications 1580:1592 modifications 1580:1592 modifications on a structural model of C9 1580:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 73 theme modifications 1580:1592 arg1 sites 1571:1575 all detected and validated sites 1544:1575 sites 1571:1575 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 7 74 theme N-glycosylation 1349:1363 arg1 site 1365:1368 a third novel, albeit low abundant, N-glycosylation site 1313:1368 a third novel, albeit low abundant, N-glycosylation site on C9 1313:1374 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 4 75 theme proteoform 580:589 arg1 profile 591:597 The proteoform profile 576:597 The proteoform profile of C9 576:603 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 6 76 theme experimental 1188:1199 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 2 77 theme attack 260:265 arg1 MAC 276:278 MAC 276:278 MAC 276:278 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 2 77 theme attack 260:265 arg1 complex 267:273 membrane attack complex 251:273 the membrane attack complex (MAC) 247:279 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 5 78 theme C9 814:815 arg1 MS/MS 874:878 MS/MS 874:878 MS/MS 874:878 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 78 theme C9 814:815 arg1 digestion 801:809 proteolytic digestion 789:809 proteolytic digestion of C9 789:815 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 78 theme C9 814:815 arg1 spectrometry 860:871 liquid chromatography (LC)-tandem mass spectrometry 821:871 liquid chromatography (LC)-tandem mass spectrometry (MS/MS) 821:879 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 7 79 gly glycosylation 1292:1304 arg2 two 1282:1284 two 1282:1284 two 1282:1284 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 7 79 gly glycosylation 1292:1304 arg2 sites 1306:1310 the two known glycosylation sites 1278:1310 the two known glycosylation sites 1278:1310 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 10 80 theme biomarker 1843:1851 arg1 discovery 1853:1861 biomarker discovery 1843:1861 biomarker discovery 1843:1861 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 0 81 theme O- 111:112 arg1 Features 95:102 Unexpected New Features 80:102 Unexpected New Features of N-, O-, and C-Glycosylation 80:133 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 3 82 theme combining 486:494 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 3 83 theme mass 519:522 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 9 84 theme C9 1619:1620 arg1 model 1610:1614 a structural model 1597:1614 a structural model of C9 1597:1620 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 7 85 theme novel 1321:1325 arg1 site 1365:1368 a third novel, albeit low abundant, N-glycosylation site 1313:1368 a third novel, albeit low abundant, N-glycosylation site on C9 1313:1374 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 10 86 theme aberrant 1867:1874 arg1 profiles 1890:1897 aberrant glycosylation profiles 1867:1897 aberrant glycosylation profiles 1867:1897 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 3 87 theme high-resolution 496:510 arg1 spectrometry 524:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 an integrative workflow combining high-resolution native mass spectrometry 462:535 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 6 88 theme proteoforms 1155:1165 arg1 specification 1087:1099 a detailed specification 1076:1099 a detailed specification 1076:1099 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 88 theme proteoforms 1155:1165 arg1 evidence 1201:1208 the first direct experimental evidence 1171:1208 the first direct experimental evidence of O-linked glycans in the N-terminal region 1171:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 88 theme proteoforms 1155:1165 arg1 validation 1102:1111 validation 1102:1111 validation 1102:1111 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 6 88 theme proteoforms 1155:1165 arg1 quantification 1118:1131 quantification 1118:1131 quantification 1118:1131 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 4 89 theme native 637:642 arg1 spectrometry 649:660 high-resolution native mass spectrometry 621:660 high-resolution native mass spectrometry 621:660 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 5 90 theme types 993:997 arg1 profiles 965:972 site-specific quantitative profiles 938:972 site-specific quantitative profiles of three different types of C9 glycosylation 938:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 90 theme types 993:997 arg1 validation 1023:1032 validation 1023:1032 validation of the native MS data 1023:1054 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 3 91 theme glyco 542:546 arg1 proteomics 564:573 (glyco)peptide-centric proteomics 541:573 (glyco)peptide-centric proteomics 541:573 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 5 92 theme mass 855:858 arg1 MS/MS 874:878 MS/MS 874:878 MS/MS 874:878 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 92 theme mass 855:858 arg1 spectrometry 860:871 liquid chromatography (LC)-tandem mass spectrometry 821:871 liquid chromatography (LC)-tandem mass spectrometry (MS/MS) 821:879 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 9 93 from present 1626:1632 arg1 MAC 1641:1643 the MAC 1637:1643 the MAC 1637:1643 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 6 94 theme co-occurring 1139:1150 arg1 proteoforms 1155:1165 15 co-occurring C9 proteoforms 1136:1165 15 co-occurring C9 proteoforms 1136:1165 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 0 95 theme C9 67:68 arg1 Mapping 19:25 Proteoform Profile Mapping 0:25 Proteoform Profile Mapping of the Human Serum Complement Component C9 0:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 5 96 theme C9 1002:1003 arg1 glycosylation 1005:1017 C9 glycosylation 1002:1017 C9 glycosylation 1002:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 97 theme liquid 821:826 arg1 LC 844:845 LC 844:845 LC 844:845 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 97 theme liquid 821:826 arg1 chromatography 828:841 liquid chromatography 821:841 liquid chromatography (LC)-tandem mass spectrometry (MS/MS) 821:879 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 7 98 theme known 1286:1290 arg1 sites 1306:1310 the two known glycosylation sites 1278:1310 the two known glycosylation sites 1278:1310 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 0 99 theme Unexpected 80:89 arg1 Features 95:102 Unexpected New Features 80:102 Unexpected New Features of N-, O-, and C-Glycosylation 80:133 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 10 100 theme pathophysiological 1924:1941 arg1 state 1943:1947 the pathophysiological state 1920:1947 the pathophysiological state of the patients 1920:1963 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 0 101 theme N- 107:108 arg1 Features 95:102 Unexpected New Features 80:102 Unexpected New Features of N-, O-, and C-Glycosylation 80:133 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 5 102 theme native 1041:1046 arg1 data 1051:1054 the native MS data 1037:1054 the native MS data 1037:1054 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 103 theme resulting 901:909 arg1 mixtures 919:926 the resulting peptide mixtures 897:926 the resulting peptide mixtures 897:926 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 4 104 theme distinct 703:710 arg1 signals 735:741 ∼50 distinct mass spectrometry (MS) signals 699:741 ∼50 distinct mass spectrometry (MS) signals 699:741 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 6 105 from quantification 1118:1131 arg1 region 1248:1253 the N-terminal region 1233:1253 the N-terminal region 1233:1253 Our study provides a detailed specification, validation, and quantification of 15 co-occurring C9 proteoforms and the first direct experimental evidence of O-linked glycans in the N-terminal region. 28221766 0 106 theme Profile 11:17 arg1 Mapping 19:25 Proteoform Profile Mapping 0:25 Proteoform Profile Mapping of the Human Serum Complement Component C9 0:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 5 107 theme data 1051:1054 arg1 profiles 965:972 site-specific quantitative profiles 938:972 site-specific quantitative profiles of three different types of C9 glycosylation 938:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 5 107 theme data 1051:1054 arg1 validation 1023:1032 validation 1023:1032 validation of the native MS data 1023:1054 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 9 108 theme pore 1679:1682 arg1 formation 1684:1692 pore formation 1679:1692 pore formation 1679:1692 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 3 109 gly microheterogeneity 398:415 arg1 C9 420:421 C9 420:421 C9 420:421 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 2 110 theme lethal 296:301 arg1 pore 303:306 a lethal pore 294:306 a lethal pore on the cellular surface of pathogenic bacteria 294:353 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 5 111 theme Subsequent 744:753 arg1 analysis 771:778 Subsequent peptide-centric analysis 744:778 Subsequent peptide-centric analysis 744:778 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 7 112 theme third 1315:1319 arg1 site 1365:1368 a third novel, albeit low abundant, N-glycosylation site 1313:1368 a third novel, albeit low abundant, N-glycosylation site on C9 1313:1374 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 2 113 theme bacteria 346:353 arg1 surface 324:330 the cellular surface 311:330 the cellular surface of pathogenic bacteria 311:353 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 0 114 theme Human 34:38 arg1 C9 67:68 the Human Serum Complement Component C9 30:68 the Human Serum Complement Component C9 30:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 9 115 from roles 1670:1674 arg1 formation 1684:1692 pore formation 1679:1692 pore formation 1679:1692 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 9 115 from roles 1670:1674 arg1 interactions 1706:1717 receptor interactions 1697:1717 receptor interactions 1697:1717 Mapping all detected and validated sites of modifications on a structural model of C9, as present in the MAC, hints at their putative roles in pore formation or receptor interactions. 28221766 5 116 theme quantitative 952:963 arg1 profiles 965:972 site-specific quantitative profiles 938:972 site-specific quantitative profiles of three different types of C9 glycosylation 938:1017 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 3 117 theme peptide-centric 548:562 arg1 proteomics 564:573 (glyco)peptide-centric proteomics 541:573 (glyco)peptide-centric proteomics 541:573 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 4 118 theme signals 735:741 arg1 co-occurrence 682:694 the co-occurrence 678:694 the co-occurrence of ∼50 distinct mass spectrometry (MS) signals 678:741 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 2 119 theme cellular 315:322 arg1 surface 324:330 the cellular surface 311:330 the cellular surface of pathogenic bacteria 311:353 It plays an essential role in the membrane attack complex (MAC), which forms a lethal pore on the cellular surface of pathogenic bacteria. 28221766 0 120 theme Complement 46:55 arg1 C9 67:68 the Human Serum Complement Component C9 30:68 the Human Serum Complement Component C9 30:68 Proteoform Profile Mapping of the Human Serum Complement Component C9 Revealing Unexpected New Features of N-, O-, and C-Glycosylation. 28221766 7 121 theme N-glycosylation 1441:1455 arg1 N-X-S/T 1466:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 the canonical N-glycosylation sequence N-X-S/T 1427:1472 Additionally, next to the two known glycosylation sites, a third novel, albeit low abundant, N-glycosylation site on C9 is identified, which surprisingly does not possess the canonical N-glycosylation sequence N-X-S/T. 28221766 5 122 theme mixtures 919:926 arg1 measurements 881:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements 789:892 proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures 789:926 Subsequent peptide-centric analysis, through proteolytic digestion of C9 and liquid chromatography (LC)-tandem mass spectrometry (MS/MS) measurements of the resulting peptide mixtures, provided site-specific quantitative profiles of three different types of C9 glycosylation and validation of the native MS data. 28221766 10 123 theme patients 1956:1963 arg1 state 1943:1947 the pathophysiological state 1920:1947 the pathophysiological state of the patients 1920:1963 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 8 124 theme Ca2+ 1519:1522 arg1 ions 1524:1527 up to two Ca2+ ions 1509:1527 up to two Ca2+ ions 1509:1527 Our data also reveal a binding of up to two Ca2+ ions to C9. 28221766 10 125 theme powerful 1759:1766 arg1 tool 1768:1771 a powerful tool 1757:1771 a powerful tool for the unbiased in-depth analysis of plasma proteins 1757:1825 The applied methods herein represent a powerful tool for the unbiased in-depth analysis of plasma proteins and may advance biomarker discovery, as aberrant glycosylation profiles may be indicative of the pathophysiological state of the patients. 28221766 4 126 theme spectrometry 717:728 arg1 signals 735:741 ∼50 distinct mass spectrometry (MS) signals 699:741 ∼50 distinct mass spectrometry (MS) signals 699:741 The proteoform profile of C9 was acquired by high-resolution native mass spectrometry, which revealed the co-occurrence of ∼50 distinct mass spectrometry (MS) signals. 28221766 3 127 theme structural 387:396 arg1 microheterogeneity 398:415 the structural microheterogeneity 383:415 the structural microheterogeneity 383:415 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28221766 3 128 theme human 437:441 arg1 serum 449:453 human blood serum 437:453 human blood serum 437:453 Here, we charted in detail the structural microheterogeneity of C9 purified from human blood serum, using an integrative workflow combining high-resolution native mass spectrometry and (glyco)peptide-centric proteomics. 28928219 4 0 theme F-box 908:912 arg1 proteins 914:921 F-box proteins 908:921 F-box proteins 908:921 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 5 1 theme Skp1 1203:1206 arg1 Skp1 1203:1206 Skp1 1203:1206 Skp1 1203:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 1 theme Skp1 1203:1206 arg1 domain 1193:1198 the intrinsically flexible F-box-binding domain 1152:1198 the intrinsically flexible F-box-binding domain of Skp1 1152:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 2 2 theme F-box-binding 588:600 arg1 domain 602:607 its C-terminal F-box-binding domain 573:607 its C-terminal F-box-binding domain 573:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 6 3 gly glycosylated 1244:1255 arg1 Skp1 1257:1260 glycosylated Skp1 1244:1260 glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data 1244:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 6 4 theme relaxation 1294:1303 arg1 kinetics 1305:1312 monosaccharide relaxation kinetics 1279:1312 monosaccharide relaxation kinetics 1279:1312 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 1 5 theme protein 263:269 arg1 substrates 271:280 protein substrates 263:280 protein substrates 263:280 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 1 6 theme F-box 176:180 arg1 proteins 182:189 F-box proteins 176:189 F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation 176:373 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 6 7 theme Skp1 1257:1260 arg1 trajectories 1228:1239 Molecular dynamics trajectories 1209:1239 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data 1209:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 2 8 attach attached 533:540 arg1 Pro-143 558:564 hydroxylated Pro-143 545:564 hydroxylated Pro-143 within its C-terminal F-box-binding domain 545:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 2 8 attach attached 533:540 arg2 pentasaccharide 517:531 a unique pentasaccharide 508:531 a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain 508:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 9 9 theme increased 1727:1735 arg1 sampling 1737:1744 The increased sampling 1723:1744 The increased sampling of an open Skp1 conformation 1723:1773 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 6 10 theme glycosylated 1244:1255 arg1 Skp1 1257:1260 glycosylated Skp1 1244:1260 glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data 1244:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 1 11 theme protein 219:225 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 9 12 theme open 1752:1755 arg1 conformation 1762:1773 an open Skp1 conformation 1749:1773 an open Skp1 conformation 1749:1773 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 3 13 from position 725:732 arg1 glycosylation 703:715 full glycosylation 698:715 full glycosylation at that position 698:732 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 4 14 with association 891:901 arg1 proteins 914:921 F-box proteins 908:921 F-box proteins 908:921 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 9 15 theme conformation 1762:1773 arg1 sampling 1737:1744 The increased sampling 1723:1744 The increased sampling of an open Skp1 conformation 1723:1773 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 6 16 theme Molecular 1209:1217 arg1 trajectories 1228:1239 Molecular dynamics trajectories 1209:1239 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data 1209:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 6 17 theme correlation 1329:1339 arg1 times 1341:1345 rotational correlation times 1318:1345 rotational correlation times 1318:1345 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 5 18 theme dynamics 1041:1048 arg1 simulations 1050:1060 NMR and molecular dynamics simulations 1023:1060 NMR and molecular dynamics simulations 1023:1060 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 3 19 theme Skp1 634:637 arg1 hydroxylation 617:629 Prolyl hydroxylation 610:629 Prolyl hydroxylation of Skp1 610:637 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 4 20 from organization 823:834 arg1 aids 876:879 aids 876:879 aids 876:879 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 4 20 from organization 823:834 arg1 Skp1 867:870 Skp1 867:870 Skp1 867:870 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 5 21 theme glycan 1110:1115 arg1 impact 1096:1101 the impact 1092:1101 the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1 1092:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 6 22 theme F-box-combining 1454:1468 arg1 site 1470:1473 the F-box-combining site 1450:1473 the F-box-combining site 1450:1473 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 1 23 theme polyubiquitin 287:299 arg1 chains 301:306 polyubiquitin chains 287:306 polyubiquitin chains that typically target them for 26S proteasome-mediated degradation 287:373 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 0 24 theme Skp1 90:93 arg1 subunit 95:101 the Dictyostelium Skp1 subunit 72:101 the Dictyostelium Skp1 subunit 72:101 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 1 25 theme proteasome-mediated 343:361 arg1 degradation 363:373 26S proteasome-mediated degradation 339:373 26S proteasome-mediated degradation 339:373 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 1 26 from proteins 182:189 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 9 27 theme F-box 1832:1836 arg1 proteins 1838:1845 F-box proteins 1832:1845 F-box proteins in cells 1832:1854 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 0 28 from site 64:67 arg1 ligases 119:125 E3 ubiquitin ligases 106:125 E3 ubiquitin ligases 106:125 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 5 29 used used 957:960 arg2 approaches 941:950 NMR and MS approaches 930:950 NMR and MS approaches 930:950 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 0 30 theme ubiquitin 109:117 arg1 ligases 119:125 E3 ubiquitin ligases 106:125 E3 ubiquitin ligases 106:125 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 5 31 theme glycan 979:984 arg1 structure 986:994 the glycan structure 975:994 the glycan structure 975:994 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 2 32 theme human 445:449 arg1 toxoplasmosis 451:463 human toxoplasmosis 445:463 human toxoplasmosis 445:463 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 5 33 theme domain 1193:1198 arg1 motions 1141:1147 motions 1141:1147 motions 1141:1147 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 33 theme domain 1193:1198 arg1 conformation 1124:1135 conformation 1124:1135 conformation 1124:1135 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 2 34 theme social 396:401 arg1 amoeba 403:408 a social amoeba 394:408 a social amoeba 394:408 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 2 34 theme social 396:401 arg1 Dictyostelium 379:391 Dictyostelium 379:391 Dictyostelium (a social amoeba) 379:409 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 7 35 theme dynamic 1570:1576 arg1 interface 1584:1592 a more accessible and dynamic F-box interface 1548:1592 a more accessible and dynamic F-box interface 1548:1592 In these trajectories, the helices separated from one another to create a more accessible and dynamic F-box interface. 28928219 5 36 dep conformation 1124:1135 arg1 the 1120:1122 the 1120:1122 the 1120:1122 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 4 37 theme Previous 770:777 arg1 studies 779:785 Previous studies 770:785 Previous studies 770:785 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 5 38 from impact 1096:1101 arg1 motions 1141:1147 motions 1141:1147 motions 1141:1147 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 38 from impact 1096:1101 arg1 conformation 1124:1135 conformation 1124:1135 conformation 1124:1135 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 39 theme F-box-binding 1179:1191 arg1 Skp1 1203:1206 Skp1 1203:1206 Skp1 1203:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 39 theme F-box-binding 1179:1191 arg1 domain 1193:1198 the intrinsically flexible F-box-binding domain 1152:1198 the intrinsically flexible F-box-binding domain of Skp1 1152:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 3 40 theme Dictyostelium 667:679 arg1 development 681:691 O2-dependent Dictyostelium development 654:691 O2-dependent Dictyostelium development 654:691 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 4 41 theme region 857:862 arg1 organization 823:834 organization 823:834 organization of the F-box-binding region in Skp1 and aids 823:879 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 5 42 theme NMR 1023:1025 arg1 simulations 1050:1060 NMR and molecular dynamics simulations 1023:1060 NMR and molecular dynamics simulations 1023:1060 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 3 43 gly glycosylation 703:715 arg1 position 725:732 that position 720:732 that position 720:732 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 3 43 gly glycosylation 703:715 arg2 position 725:732 that position 720:732 that position 720:732 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 3 44 theme full 698:701 arg1 glycosylation 703:715 full glycosylation 698:715 full glycosylation at that position 698:732 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 8 45 theme disordered 1679:1688 arg1 region 1690:1695 a disordered region 1677:1695 a disordered region of a full-length protein 1677:1720 These results offer an unprecedented view of how a glycan modification influences a disordered region of a full-length protein. 28928219 1 46 theme ubiquitin 231:239 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 6 47 theme rotational 1318:1327 arg1 times 1341:1345 rotational correlation times 1318:1345 rotational correlation times 1318:1345 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 8 48 theme full-length 1702:1712 arg1 protein 1714:1720 a full-length protein 1700:1720 a full-length protein 1700:1720 These results offer an unprecedented view of how a glycan modification influences a disordered region of a full-length protein. 28928219 8 49 theme glycan 1646:1651 arg1 modification 1653:1664 a glycan modification 1644:1664 a glycan modification 1644:1664 These results offer an unprecedented view of how a glycan modification influences a disordered region of a full-length protein. 28928219 2 50 theme C-terminal 577:586 arg1 domain 602:607 its C-terminal F-box-binding domain 573:607 its C-terminal F-box-binding domain 573:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 3 51 theme optimal 750:756 arg1 sensing 761:767 optimal O2 sensing 750:767 optimal O2 sensing 750:767 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 6 52 theme monosaccharide 1279:1292 arg1 kinetics 1305:1312 monosaccharide relaxation kinetics 1279:1312 monosaccharide relaxation kinetics 1279:1312 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 2 53 dep gondii 423:428 arg1 agent 435:439 the agent 431:439 the agent for human toxoplasmosis 431:463 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 1 54 theme SCF 194:196 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 6 55 theme NMR 1363:1365 arg1 data 1367:1370 the NMR data 1359:1370 the NMR data 1359:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 1 56 theme Skp1/Cullin-1/F-box 199:217 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 2 57 theme unique 510:515 arg1 pentasaccharide 517:531 a unique pentasaccharide 508:531 a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain 508:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 9 58 with interactions 1814:1825 arg1 proteins 1838:1845 F-box proteins 1832:1845 F-box proteins in cells 1832:1854 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 9 59 theme Skp1 1757:1760 arg1 conformation 1762:1773 an open Skp1 conformation 1749:1773 an open Skp1 conformation 1749:1773 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 6 60 theme dynamics 1219:1226 arg1 trajectories 1228:1239 Molecular dynamics trajectories 1209:1239 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data 1209:1370 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 5 61 theme molecular 1031:1039 arg1 dynamics 1041:1048 molecular dynamics 1031:1048 molecular dynamics 1031:1048 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 62 theme NMR 930:932 arg1 approaches 941:950 NMR and MS approaches 930:950 NMR and MS approaches 930:950 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 2 63 theme hydroxylated 545:556 arg1 Pro-143 558:564 hydroxylated Pro-143 545:564 hydroxylated Pro-143 within its C-terminal F-box-binding domain 545:607 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 0 64 theme Dictyostelium 76:88 arg1 subunit 95:101 the Dictyostelium Skp1 subunit 72:101 the Dictyostelium Skp1 subunit 72:101 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 5 65 theme simulations 1050:1060 arg1 combination 1008:1018 then a combination 1001:1018 then a combination of NMR and molecular dynamics simulations 1001:1060 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 3 66 theme Prolyl 610:615 arg1 hydroxylation 617:629 Prolyl hydroxylation 610:629 Prolyl hydroxylation of Skp1 610:637 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 1 67 theme conserved 138:146 arg1 protein 148:154 a conserved protein 136:154 a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation 136:373 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 1 67 theme conserved 138:146 arg1 Skp1 128:131 Skp1 128:131 Skp1 128:131 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 0 68 theme subunit 95:101 arg1 subunit 95:101 the Dictyostelium Skp1 subunit 72:101 the Dictyostelium Skp1 subunit 72:101 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 0 68 theme subunit 95:101 arg1 site 64:67 the F-box-combining site 44:67 the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases 44:125 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 1 69 theme 26S 339:341 arg1 degradation 363:373 26S proteasome-mediated degradation 339:373 26S proteasome-mediated degradation 339:373 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 2 70 theme other 471:475 arg1 protists 477:484 other protists 471:484 other protists 471:484 In Dictyostelium (a social amoeba), Toxoplasma gondii (the agent for human toxoplasmosis), and other protists, Skp1 is regulated by a unique pentasaccharide attached to hydroxylated Pro-143 within its C-terminal F-box-binding domain. 28928219 0 71 theme E3 106:107 arg1 ligases 119:125 E3 ubiquitin ligases 106:125 E3 ubiquitin ligases 106:125 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 7 72 theme F-box 1578:1582 arg1 interface 1584:1592 a more accessible and dynamic F-box interface 1548:1592 a more accessible and dynamic F-box interface 1548:1592 In these trajectories, the helices separated from one another to create a more accessible and dynamic F-box interface. 28928219 0 73 theme sensing-associated 3:20 arg1 glycosylation 22:34 O2 sensing-associated glycosylation 0:34 O2 sensing-associated glycosylation 0:34 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 5 74 theme MS 938:939 arg1 approaches 941:950 NMR and MS approaches 930:950 NMR and MS approaches 930:950 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 6 75 theme site 1470:1473 arg1 α-helices 1437:1445 two α-helices 1433:1445 two α-helices of the F-box-combining site 1433:1473 Molecular dynamics trajectories of glycosylated Skp1 whose calculated monosaccharide relaxation kinetics and rotational correlation times agreed with the NMR data indicated that the glycan interacts with the loop connecting two α-helices of the F-box-combining site. 28928219 5 76 theme flexible 1170:1177 arg1 Skp1 1203:1206 Skp1 1203:1206 Skp1 1203:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 5 76 theme flexible 1170:1177 arg1 domain 1193:1198 the intrinsically flexible F-box-binding domain 1152:1198 the intrinsically flexible F-box-binding domain of Skp1 1152:1206 Here, NMR and MS approaches were used to determine the glycan structure, and then a combination of NMR and molecular dynamics simulations were employed to characterize the impact of the glycan on the conformation and motions of the intrinsically flexible F-box-binding domain of Skp1. 28928219 3 77 theme O2-dependent 654:665 arg1 development 681:691 O2-dependent Dictyostelium development 654:691 O2-dependent Dictyostelium development 654:691 Prolyl hydroxylation of Skp1 contributes to O2-dependent Dictyostelium development, but full glycosylation at that position is required for optimal O2 sensing. 28928219 0 78 theme F-box-combining 48:62 arg1 subunit 95:101 the Dictyostelium Skp1 subunit 72:101 the Dictyostelium Skp1 subunit 72:101 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 0 78 theme F-box-combining 48:62 arg1 site 64:67 the F-box-combining site 44:67 the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases 44:125 O2 sensing-associated glycosylation exposes the F-box-combining site of the Dictyostelium Skp1 subunit in E3 ubiquitin ligases. 28928219 7 79 dep one 1526:1528 arg1 another 1530:1536 another 1530:1536 another 1530:1536 In these trajectories, the helices separated from one another to create a more accessible and dynamic F-box interface. 28928219 9 80 from proteins 1838:1845 arg1 cells 1850:1854 cells 1850:1854 cells 1850:1854 The increased sampling of an open Skp1 conformation can explain how glycosylation enhances interactions with F-box proteins in cells. 28928219 7 81 theme accessible 1555:1564 arg1 interface 1584:1592 a more accessible and dynamic F-box interface 1548:1592 a more accessible and dynamic F-box interface 1548:1592 In these trajectories, the helices separated from one another to create a more accessible and dynamic F-box interface. 28928219 8 82 theme unprecedented 1618:1630 arg1 view 1632:1635 an unprecedented view 1615:1635 an unprecedented view of how a glycan modification influences a disordered region of a full-length protein 1615:1720 These results offer an unprecedented view of how a glycan modification influences a disordered region of a full-length protein. 28928219 4 83 theme F-box-binding 843:855 arg1 region 857:862 the F-box-binding region 839:862 the F-box-binding region 839:862 Previous studies have shown that the glycan promotes organization of the F-box-binding region in Skp1 and aids in Skp1's association with F-box proteins. 28928219 1 84 theme E3 228:229 arg1 ligases 241:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases 194:247 Skp1 is a conserved protein linking cullin-1 to F-box proteins in SCF (Skp1/Cullin-1/F-box protein) E3 ubiquitin ligases, which modify protein substrates with polyubiquitin chains that typically target them for 26S proteasome-mediated degradation. 28928219 8 85 theme protein 1714:1720 arg1 region 1690:1695 a disordered region 1677:1695 a disordered region of a full-length protein 1677:1720 These results offer an unprecedented view of how a glycan modification influences a disordered region of a full-length protein. 25759508 6 0 with CE-MS/MS 835:842 arg1 fragmentation 853:865 CID fragmentation 849:865 CID fragmentation 849:865 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 4 1 from threonine 606:614 arg1 O-glycosylated 584:597 O-glycosylated 584:597 O-glycosylated 584:597 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 1 from threonine 606:614 arg1 regions 529:535 The repeat regions 518:535 The repeat regions within the IgG3 hinge 518:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 1 from threonine 606:614 arg1 motif 637:641 the triple repeat motif 619:641 the triple repeat motif 619:641 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 2 2 theme amount 230:235 arg1 amount 230:235 the total amount 220:235 the total amount of IgG 220:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 2 2 theme amount 230:235 arg1 % 215:215 ∼8% 213:215 ∼8% of the total amount of IgG 213:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 2 2 theme amount 230:235 arg1 IgG 240:242 IgG 240:242 IgG 240:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 8 3 theme Fc 1237:1238 arg1 domain 1244:1249 the Fc CH2 domain 1233:1249 the Fc CH2 domain 1233:1249 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 4 4 from part 579:582 arg1 regions 529:535 The repeat regions 518:535 The repeat regions within the IgG3 hinge 518:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 4 from part 579:582 arg1 O-glycosylated 584:597 O-glycosylated 584:597 O-glycosylated 584:597 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 8 5 theme O-glycan 1129:1136 arg1 structures 1138:1147 similar O-glycan structures 1121:1147 similar O-glycan structures 1121:1147 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 6 6 theme dissociation 804:815 arg1 fragmentation 817:829 dissociation fragmentation 804:829 dissociation fragmentation 804:829 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 7 7 gly O-glycosylation 1035:1049 arg2 sites 1051:1055 potential O-glycosylation sites 1025:1055 potential O-glycosylation sites 1025:1055 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 7 8 theme O-glycosylation 1035:1049 arg1 sites 1051:1055 potential O-glycosylation sites 1025:1055 potential O-glycosylation sites 1025:1055 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 2 9 theme human 247:251 arg1 serum 253:257 human serum 247:257 human serum 247:257 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 9 10 theme IgG3 1397:1400 arg1 samples 1402:1408 six serum-derived IgG3 samples 1379:1408 six serum-derived IgG3 samples 1379:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 6 11 theme IgG3 902:905 arg1 O-glycosylation 907:921 IgG3 O-glycosylation 902:921 IgG3 O-glycosylation 902:921 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 8 12 theme conserved 1204:1212 arg1 N-glycosylation 1214:1228 the conserved N-glycosylation 1200:1228 the conserved N-glycosylation of the Fc CH2 domain 1200:1249 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 2 13 theme IgG 240:242 arg1 amount 230:235 the total amount 220:235 the total amount of IgG 220:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 2 13 theme IgG 240:242 arg1 IgG 240:242 IgG 240:242 IgG 240:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 7 14 theme IgG3 1011:1014 arg1 production 990:999 the recombinant production 974:999 the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out 974:1076 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 4 15 theme repeat 630:635 arg1 motif 637:641 the triple repeat motif 619:641 the triple repeat motif 619:641 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 5 16 dep Non- 644:647 arg1 mono- 650:654 mono- 650:654 mono- 650:654 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 5 16 dep Non- 644:647 arg1 disialylated 660:671 disialylated 660:671 disialylated 660:671 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 8 17 theme domain 1244:1249 arg1 N-glycosylation 1214:1228 the conserved N-glycosylation 1200:1228 the conserved N-glycosylation of the Fc CH2 domain 1200:1249 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 7 18 theme recombinant 978:988 arg1 production 990:999 the recombinant production 974:999 the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out 974:1076 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 8 19 gly N-glycosylation 1214:1228 arg1 domain 1244:1249 the Fc CH2 domain 1233:1249 the Fc CH2 domain 1233:1249 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 7 20 theme O-glycosylation 928:942 arg1 site 944:947 The O-glycosylation site 924:947 The O-glycosylation site 924:947 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 9 21 theme IgG3 1437:1440 arg1 allotypes 1442:1450 two monoclonal IgG3 allotypes 1422:1450 two monoclonal IgG3 allotypes 1422:1450 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 2 22 theme enhanced 346:353 arg1 functions 364:372 enhanced effector functions 346:372 enhanced effector functions 346:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 8 23 theme considerable 1258:1269 arg1 variation 1287:1295 considerable interindividual variation 1258:1295 considerable interindividual variation 1258:1295 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 5 24 theme core 673:676 arg1 O-glycans 685:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 1 25 theme proteins 115:122 arg1 proteins 115:122 the most abundant proteins 97:122 the most abundant proteins present in human serum and a fundamental component of the immune system 97:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 25 theme proteins 115:122 arg1 one 90:92 one 90:92 one 90:92 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 8 26 dep found 1115:1119 arg1 whereas 1171:1177 whereas 1171:1177 whereas 1171:1177 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 9 27 from % 1374:1374 arg1 allotypes 1442:1450 two monoclonal IgG3 allotypes 1422:1450 two monoclonal IgG3 allotypes 1422:1450 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 9 27 from % 1374:1374 arg1 samples 1402:1408 six serum-derived IgG3 samples 1379:1408 six serum-derived IgG3 samples 1379:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 1 28 from present 124:130 arg1 component 165:173 a fundamental component 151:173 a fundamental component of the immune system 151:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 28 from present 124:130 arg1 serum 141:145 human serum 135:145 human serum 135:145 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 2 29 theme elongated 319:327 arg1 region 335:340 its elongated hinge region 315:340 its elongated hinge region 315:340 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 5 30 located detected 700:707 arg2 O-glycans 685:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 5 30 located detected 700:707 arg1 samples 725:731 various IgG3 samples 712:731 various IgG3 samples 712:731 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 6 31 with NanoLC-ESI-IT-MS/MS 761:779 arg1 CE-MS/MS 835:842 CE-MS/MS 835:842 CE-MS/MS with CID fragmentation 835:865 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 6 31 with NanoLC-ESI-IT-MS/MS 761:779 arg1 transfer 795:802 electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation 786:865 transfer 795:802 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 0 32 theme Hinge-Region 0:11 arg1 O-Glycosylation 13:27 Hinge-Region O-Glycosylation 0:27 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 0:62 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 2 33 theme functions 364:372 arg1 subclasses 293:302 the other IgG subclasses 279:302 the other IgG subclasses because of its elongated hinge region and enhanced effector functions 279:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 7 34 gly O-glycosylation 928:942 arg2 site 944:947 The O-glycosylation site 924:947 The O-glycosylation site 924:947 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 3 35 theme partial 394:400 arg1 O-glycosylation 402:416 partial O-glycosylation 394:416 partial O-glycosylation 394:416 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 2 36 theme total 224:228 arg1 amount 230:235 the total amount 220:235 the total amount of IgG 220:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 2 36 theme total 224:228 arg1 IgG 240:242 IgG 240:242 IgG 240:242 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 3 37 gly O-glycosylation 402:416 arg1 region 436:441 the IgG3 hinge region 421:441 the IgG3 hinge region 421:441 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 2 38 theme region 335:340 arg1 subclasses 293:302 the other IgG subclasses 279:302 the other IgG subclasses because of its elongated hinge region and enhanced effector functions 279:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 0 39 theme Immunoglobulin 38:51 arg1 G3 53:54 Human Immunoglobulin G3 32:54 Human Immunoglobulin G3 (IgG3) 32:61 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 0 39 theme Immunoglobulin 38:51 arg1 IgG3 57:60 IgG3 57:60 IgG3 57:60 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 9 40 theme each 1315:1318 arg1 occupancy 1302:1310 The occupancy 1298:1310 The occupancy of each of the three O-glycosylation sites 1298:1353 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 9 40 theme each 1315:1318 arg1 % 1374:1374 ∼10% 1371:1374 ∼10% in six serum-derived IgG3 samples 1371:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 0 41 gly O-Glycosylation 13:27 arg1 G3 53:54 Human Immunoglobulin G3 32:54 Human Immunoglobulin G3 (IgG3) 32:61 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 0 41 gly O-Glycosylation 13:27 arg1 IgG3 57:60 IgG3 57:60 IgG3 57:60 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 3 42 theme IgG3 425:428 arg1 region 436:441 the IgG3 hinge region 421:441 the IgG3 hinge region 421:441 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 5 43 theme various 712:718 arg1 samples 725:731 various IgG3 samples 712:731 various IgG3 samples 712:731 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 1 44 theme immune 182:187 arg1 system 189:194 the immune system 178:194 the immune system 178:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 8 45 from donors 1105:1110 arg1 samples 1088:1094 IgG3 samples 1083:1094 IgG3 samples from six donors 1083:1110 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 4 46 theme IgG3 548:551 arg1 hinge 553:557 the IgG3 hinge 544:557 the IgG3 hinge 544:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 2 47 theme other 283:287 arg1 subclasses 293:302 the other IgG subclasses 279:302 the other IgG subclasses because of its elongated hinge region and enhanced effector functions 279:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 6 48 theme CID 849:851 arg1 fragmentation 853:865 CID fragmentation 849:865 CID fragmentation 849:865 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 4 49 gly O-glycosylated 584:597 arg1 regions 529:535 The repeat regions 518:535 The repeat regions within the IgG3 hinge 518:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 49 gly O-glycosylated 584:597 arg2 O-glycosylated 584:597 O-glycosylated 584:597 O-glycosylated 584:597 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 49 gly O-glycosylated 584:597 arg2 regions 529:535 The repeat regions 518:535 The repeat regions within the IgG3 hinge 518:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 49 gly O-glycosylated 584:597 arg1 part 579:582 part 579:582 part 579:582 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 49 gly O-glycosylated 584:597 arg1 O-glycosylated 584:597 O-glycosylated 584:597 O-glycosylated 584:597 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 49 gly O-glycosylated 584:597 arg2 threonine 606:614 the threonine 602:614 the threonine in the triple repeat motif 602:641 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 3 50 theme /MS 475:477 arg1 analysis 480:487 nanoLC-ESI-IT-MS(/MS) analysis 458:487 nanoLC-ESI-IT-MS(/MS) analysis 458:487 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 4 51 theme repeat 522:527 arg1 regions 529:535 The repeat regions 518:535 The repeat regions within the IgG3 hinge 518:557 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 4 51 theme repeat 522:527 arg1 O-glycosylated 584:597 O-glycosylated 584:597 O-glycosylated 584:597 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 3 52 theme nanoLC-ESI-IT-MS 458:473 arg1 analysis 480:487 nanoLC-ESI-IT-MS(/MS) analysis 458:487 nanoLC-ESI-IT-MS(/MS) analysis 458:487 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 4 53 from O-glycosylated 584:597 arg1 part 579:582 part 579:582 part 579:582 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 1 54 theme human 135:139 arg1 serum 141:145 human serum 135:145 human serum 135:145 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 55 attach present 124:130 arg1 component 165:173 a fundamental component 151:173 a fundamental component of the immune system 151:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 55 attach present 124:130 arg2 proteins 115:122 the most abundant proteins 97:122 the most abundant proteins present in human serum and a fundamental component of the immune system 97:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 55 attach present 124:130 arg1 serum 141:145 human serum 135:145 human serum 135:145 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 7 56 theme potential 1025:1033 arg1 sites 1051:1055 potential O-glycosylation sites 1025:1055 potential O-glycosylation sites 1025:1055 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 6 57 gly O-glycosylation 907:921 arg2 O-glycosylation 907:921 IgG3 O-glycosylation 902:921 IgG3 O-glycosylation 902:921 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 6 57 gly O-glycosylation 907:921 arg2 site 894:897 the site 890:897 the site of IgG3 O-glycosylation 890:921 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 3 58 theme proteolytic 495:505 arg1 digestion 507:515 proteolytic digestion 495:515 proteolytic digestion 495:515 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 9 59 theme serum-derived 1383:1395 arg1 samples 1402:1408 six serum-derived IgG3 samples 1379:1408 six serum-derived IgG3 samples 1379:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 8 60 theme similar 1121:1127 arg1 structures 1138:1147 similar O-glycan structures 1121:1147 similar O-glycan structures 1121:1147 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 6 61 theme O-glycosylation 907:921 arg1 site 894:897 the site 890:897 the site of IgG3 O-glycosylation 890:921 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 6 61 theme O-glycosylation 907:921 arg1 O-glycosylation 907:921 IgG3 O-glycosylation 902:921 IgG3 O-glycosylation 902:921 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 8 62 theme same 1187:1190 arg1 samples 1192:1198 the same samples 1183:1198 the same samples 1183:1198 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 6 63 used used 872:875 arg2 NanoLC-ESI-IT-MS/MS 761:779 NanoLC-ESI-IT-MS/MS 761:779 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation 761:865 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 5 64 theme poly- 739:743 arg1 samples 725:731 various IgG3 samples 712:731 various IgG3 samples 712:731 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 3 65 theme region 436:441 arg1 O-glycosylation 402:416 partial O-glycosylation 394:416 partial O-glycosylation 394:416 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 7 66 theme mutant 1004:1009 arg1 IgG3 1011:1014 mutant IgG3 1004:1014 mutant IgG3 in which potential O-glycosylation sites had been knocked out 1004:1076 The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out. 25759508 5 67 theme monoclonal 749:758 arg1 samples 725:731 various IgG3 samples 712:731 various IgG3 samples 712:731 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 8 68 theme site 1153:1156 arg1 occupancies 1158:1168 site occupancies 1153:1168 site occupancies 1153:1168 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 4 69 theme triple 623:628 arg1 motif 637:641 the triple repeat motif 619:641 the triple repeat motif 619:641 The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. 25759508 1 70 from component 165:173 arg1 present 124:130 present 124:130 present 124:130 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 9 71 theme monoclonal 1426:1435 arg1 allotypes 1442:1450 two monoclonal IgG3 allotypes 1422:1450 two monoclonal IgG3 allotypes 1422:1450 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 8 72 theme CH2 1240:1242 arg1 domain 1244:1249 the Fc CH2 domain 1233:1249 the Fc CH2 domain 1233:1249 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 1 73 theme fundamental 153:163 arg1 component 165:173 a fundamental component 151:173 a fundamental component of the immune system 151:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 74 theme abundant 106:113 arg1 proteins 115:122 the most abundant proteins 97:122 the most abundant proteins present in human serum and a fundamental component of the immune system 97:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 9 75 theme O-glycosylation 1333:1347 arg1 sites 1349:1353 the three O-glycosylation sites 1323:1353 the three O-glycosylation sites 1323:1353 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 5 76 theme 1-type 678:683 arg1 O-glycans 685:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 1 77 from serum 141:145 arg1 present 124:130 present 124:130 present 124:130 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 78 theme present 124:130 arg1 proteins 115:122 the most abundant proteins 97:122 the most abundant proteins present in human serum and a fundamental component of the immune system 97:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 8 79 theme interindividual 1271:1285 arg1 variation 1287:1295 considerable interindividual variation 1258:1295 considerable interindividual variation 1258:1295 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 5 80 theme Non- 644:647 arg1 O-glycans 685:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans 644:693 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 9 81 link serum-derived 1383:1395 arg1 samples 1402:1408 six serum-derived IgG3 samples 1379:1408 six serum-derived IgG3 samples 1379:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 1 82 theme Immunoglobulin 64:77 arg1 IgG 82:84 IgG 82:84 IgG 82:84 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 82 theme Immunoglobulin 64:77 arg1 G 79:79 Immunoglobulin G 64:79 Immunoglobulin G (IgG) 64:85 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 9 83 gly occupancy 1302:1310 arg2 each 1315:1318 each 1315:1318 each 1315:1318 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 9 83 gly occupancy 1302:1310 arg2 sites 1349:1353 the three O-glycosylation sites 1323:1353 the three O-glycosylation sites 1323:1353 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 5 84 theme IgG3 720:723 arg1 samples 725:731 various IgG3 samples 712:731 various IgG3 samples 712:731 Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. 25759508 6 85 with transfer 795:802 arg1 fragmentation 853:865 CID fragmentation 849:865 CID fragmentation 849:865 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 0 86 theme Human 32:36 arg1 G3 53:54 Human Immunoglobulin G3 32:54 Human Immunoglobulin G3 (IgG3) 32:61 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 0 86 theme Human 32:36 arg1 IgG3 57:60 IgG3 57:60 IgG3 57:60 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 2 87 theme effector 355:362 arg1 functions 364:372 enhanced effector functions 346:372 enhanced effector functions 346:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 0 88 theme G3 53:54 arg1 O-Glycosylation 13:27 Hinge-Region O-Glycosylation 0:27 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 0:62 Hinge-Region O-Glycosylation of Human Immunoglobulin G3 (IgG3). 25759508 6 89 theme electron 786:793 arg1 transfer 795:802 electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation 786:865 transfer 795:802 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 9 90 gly O-glycosylation 1333:1347 arg2 three 1327:1331 three 1327:1331 three 1327:1331 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 9 90 gly O-glycosylation 1333:1347 arg2 sites 1349:1353 the three O-glycosylation sites 1323:1353 the three O-glycosylation sites 1323:1353 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 2 91 theme hinge 329:333 arg1 region 335:340 its elongated hinge region 315:340 its elongated hinge region 315:340 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 25759508 9 92 from % 1417:1417 arg1 allotypes 1442:1450 two monoclonal IgG3 allotypes 1422:1450 two monoclonal IgG3 allotypes 1422:1450 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 9 92 from % 1417:1417 arg1 samples 1402:1408 six serum-derived IgG3 samples 1379:1408 six serum-derived IgG3 samples 1379:1408 The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes. 25759508 6 93 dep transfer 795:802 arg1 fragmentation 817:829 dissociation fragmentation 804:829 dissociation fragmentation 804:829 NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. 25759508 3 94 theme hinge 430:434 arg1 region 436:441 the IgG3 hinge region 421:441 the IgG3 hinge region 421:441 This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. 25759508 1 95 theme system 189:194 arg1 component 165:173 a fundamental component 151:173 a fundamental component of the immune system 151:194 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 1 95 theme system 189:194 arg1 serum 141:145 human serum 135:145 human serum 135:145 Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. 25759508 8 96 theme IgG3 1083:1086 arg1 samples 1088:1094 IgG3 samples 1083:1094 IgG3 samples from six donors 1083:1110 For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. 25759508 2 97 theme IgG 289:291 arg1 subclasses 293:302 the other IgG subclasses 279:302 the other IgG subclasses because of its elongated hinge region and enhanced effector functions 279:372 IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. 26202417 0 0 theme Added 93:97 arg1 Sites 135:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 7 1 theme MALDI-TOF 919:927 arg1 MS 929:930 MALDI-TOF MS 919:930 MALDI-TOF MS 919:930 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 8 2 dep class 1080:1084 arg1 the 1076:1078 the 1076:1078 the 1076:1078 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 9 3 theme immune 1388:1393 arg1 responses 1402:1410 the immune system responses 1384:1410 the immune system responses 1384:1410 Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation. 26202417 8 4 theme SP-D 1247:1250 arg1 collectin 1257:1265 HA and SP-D lung collectin 1240:1265 HA and SP-D lung collectin 1240:1265 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 0 5 theme Relevant 112:119 arg1 Sites 135:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 6 6 theme engineered 772:781 arg1 forms 783:787 engineered forms 772:787 engineered forms 772:787 HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. 26202417 9 7 theme system 1395:1400 arg1 responses 1402:1410 the immune system responses 1384:1410 the immune system responses 1384:1410 Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation. 26202417 6 8 dep studied 747:753 arg1 include 755:761 include 755:761 include HK68 and engineered forms with 1, 2, and 4 added sites 755:816 HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. 26202417 0 9 theme Historically 99:110 arg1 Sites 135:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 8 10 dep varies 1116:1121 arg1 2 1073:1073 2 1073:1073 2 1073:1073 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 3 11 gly N-glycosylation 396:410 arg2 sequons 412:418 N-glycosylation sequons 396:418 N-glycosylation sequons 396:418 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 7 12 theme site 890:893 arg1 analysis 905:912 sequence and site occupancy analysis 877:912 analysis 905:912 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 4 13 theme glycosylation 468:480 arg1 expansion 482:490 This glycosylation expansion 463:490 This glycosylation expansion 463:490 This glycosylation expansion has not been studied at the structural level. 26202417 0 14 with Hemagglutinins 60:73 arg1 Sites 135:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 0 15 theme Glycosylation 121:133 arg1 Sites 135:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Sequentially Added Historically Relevant Glycosylation Sites 80:139 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 8 16 used occupied 1055:1062 arg2 majority 1029:1036 the majority 1025:1036 the majority of N-sequons 1025:1049 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 7 17 theme sequence 877:884 arg1 analysis 905:912 sequence and site occupancy analysis 877:912 analysis 905:912 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 8 18 dep occupied 1055:1062 arg1 1 1022:1022 1 1022:1022 1 1022:1022 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 19 theme high 1284:1287 arg1 Asn 1172:1174 Asn 165 1172:1178 Asn 165 1172:1178 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 19 theme high 1284:1287 arg1 Asn 1184:1186 Asn 246 1184:1190 Asn 246 1184:1190 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 19 theme high 1284:1287 arg1 type 1297:1300 exclusively high mannose type 1272:1300 exclusively high mannose type 1272:1300 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 5 20 theme H3N2 576:579 arg1 strains 587:593 H3N2 virus strains 576:593 H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 576:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 9 21 theme previous 1327:1334 arg1 reports 1336:1342 previous reports 1327:1342 previous reports 1327:1342 Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation. 26202417 0 22 theme Glycosylation 0:12 arg1 Analysis 14:21 Glycosylation Analysis 0:21 Glycosylation Analysis of Engineered H3N2 Influenza A Virus 0:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 5 23 theme glycosylation 636:648 arg1 sites 650:654 glycosylation sites 636:654 glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 636:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 5 24 theme virus 581:585 arg1 strains 587:593 H3N2 virus strains 576:593 H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 576:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 7 25 gly glycopeptide 851:862 arg2 glycopeptide 851:862 glycopeptide composition 851:874 glycopeptide composition 851:874 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 0 26 theme Engineered 26:35 arg1 Virus 54:58 Engineered H3N2 Influenza A Virus 26:58 Engineered H3N2 Influenza A Virus 26:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 3 27 dep a 338:338 arg1 leap 340:343 leap 340:343 leap to humans from a zoonotic host 340:374 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 5 28 theme strains 587:593 arg1 N-glycosylation 557:571 HA N-glycosylation 554:571 HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 554:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 8 29 theme lung 1252:1255 arg1 collectin 1257:1265 HA and SP-D lung collectin 1240:1265 HA and SP-D lung collectin 1240:1265 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 1 30 theme major 209:213 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 30 theme major 209:213 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 7 31 theme permethylation 932:945 arg1 profiling 947:955 permethylation profiling 932:955 permethylation profiling 932:955 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 5 32 gly N-glycosylation 557:571 arg1 strains 587:593 H3N2 virus strains 576:593 H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 576:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 2 33 theme antigenic 305:313 arg1 characteristics 315:329 HA's antigenic characteristics 300:329 HA's antigenic characteristics 300:329 The oligosaccharides of HA can contribute to HA's antigenic characteristics. 26202417 7 34 theme occupancy 895:903 arg1 analysis 905:912 sequence and site occupancy analysis 877:912 analysis 905:912 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 8 35 theme proteins 1159:1166 arg1 landscape 1142:1150 the landscape 1138:1150 the landscape of the proteins 1138:1166 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 0 36 theme Influenza 42:50 arg1 Virus 54:58 Engineered H3N2 Influenza A Virus 26:58 Engineered H3N2 Influenza A Virus 26:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 9 37 theme structural 1355:1364 arg1 insight 1366:1372 structural insight 1355:1372 structural insight as to how the immune system responses may differ depending on HA glycosylation 1355:1451 Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation. 26202417 5 38 gly glycosylation 636:648 arg2 sites 650:654 glycosylation sites 636:654 glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 636:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 3 39 theme fitness 445:451 arg1 strategy 453:460 its fitness strategy 441:460 its fitness strategy 441:460 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 0 40 theme H3N2 37:40 arg1 Virus 54:58 Engineered H3N2 Influenza A Virus 26:58 Engineered H3N2 Influenza A Virus 26:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 6 41 with HK68 763:766 arg1 sites 812:816 1, 2, and 4 added sites 794:816 1, 2, and 4 added sites 794:816 HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. 26202417 8 42 theme glycans 1108:1114 arg1 complexity 1090:1099 complexity 1090:1099 complexity 1090:1099 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 42 theme glycans 1108:1114 arg1 class 1080:1084 class 1080:1084 class 1080:1084 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 1 43 theme host 225:228 arg1 antibodies 243:252 host neutralizing antibodies 225:252 host neutralizing antibodies 225:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 0 44 theme Virus 54:58 arg1 Analysis 14:21 Glycosylation Analysis 0:21 Glycosylation Analysis of Engineered H3N2 Influenza A Virus 0:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 3 45 theme zoonotic 362:369 arg1 host 371:374 a zoonotic host 360:374 a zoonotic host 360:374 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 1 46 gly glycoprotein 170:181 arg1 glycoprotein 170:181 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 6 47 with forms 783:787 arg1 sites 812:816 1, 2, and 4 added sites 794:816 1, 2, and 4 added sites 794:816 HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. 26202417 8 48 theme HA 1240:1241 arg1 collectin 1257:1265 HA and SP-D lung collectin 1240:1265 HA and SP-D lung collectin 1240:1265 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 0 49 theme A 52:52 arg1 Virus 54:58 Engineered H3N2 Influenza A Virus 26:58 Engineered H3N2 Influenza A Virus 26:58 Glycosylation Analysis of Engineered H3N2 Influenza A Virus Hemagglutinins with Sequentially Added Historically Relevant Glycosylation Sites. 26202417 8 50 theme N-sequons 1041:1049 arg1 majority 1029:1036 the majority 1025:1036 the majority of N-sequons 1025:1049 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 6 51 theme added 806:810 arg1 sites 812:816 1, 2, and 4 added sites 794:816 1, 2, and 4 added sites 794:816 HAs studied include HK68 and engineered forms with 1, 2, and 4 added sites. 26202417 5 52 theme HA 554:555 arg1 N-glycosylation 557:571 HA N-glycosylation 554:571 HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68) 554:740 Here we examine HA N-glycosylation of H3N2 virus strains that we have engineered to closely mimic glycosylation sites gained between 1968 through 2002 starting with pandemic A/Hong Kong/1/68 (H3N2: HK68). 26202417 8 53 dep type 1297:1300 arg1 3 1169:1169 3 1169:1169 3 1169:1169 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 7 54 theme released 981:988 arg1 glycans 990:996 released glycans 981:996 released glycans 981:996 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 8 55 theme mannose 1289:1295 arg1 Asn 1172:1174 Asn 165 1172:1178 Asn 165 1172:1178 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 55 theme mannose 1289:1295 arg1 Asn 1184:1186 Asn 246 1184:1190 Asn 246 1184:1190 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 8 55 theme mannose 1289:1295 arg1 type 1297:1300 exclusively high mannose type 1272:1300 exclusively high mannose type 1272:1300 Our study reveals that 1) the majority of N-sequons are occupied at ≥90%, 2) the class and complexity of the glycans varies by region over the landscape of the proteins, 3) Asn 165 and Asn 246, which are associated with interactions between HA and SP-D lung collectin, are exclusively high mannose type. 26202417 1 56 theme influenza 146:154 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 56 theme influenza 146:154 arg1 HA 198:199 HA 198:199 HA 198:199 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 56 theme influenza 146:154 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 7 57 theme glycopeptide 851:862 arg1 composition 864:874 glycopeptide composition 851:874 glycopeptide composition 851:874 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 1 58 theme neutralizing 230:241 arg1 antibodies 243:252 host neutralizing antibodies 225:252 host neutralizing antibodies 225:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 7 59 theme glycans 990:996 arg1 characterization 961:976 characterization 961:976 characterization of released glycans 961:996 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 2 60 theme HA 279:280 arg1 oligosaccharides 259:274 The oligosaccharides 255:274 The oligosaccharides of HA 255:280 The oligosaccharides of HA can contribute to HA's antigenic characteristics. 26202417 1 61 theme virus 156:160 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 61 theme virus 156:160 arg1 HA 198:199 HA 198:199 HA 198:199 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 61 theme virus 156:160 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 62 theme antibodies 243:252 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 62 theme antibodies 243:252 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 3 63 theme strategy 453:460 arg1 part 433:436 part 433:436 part of its fitness strategy 433:460 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 7 64 used used 827:830 arg2 We 819:820 We 819:820 We 819:820 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 1 65 theme surface 162:168 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 65 theme surface 162:168 arg1 HA 198:199 HA 198:199 HA 198:199 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 65 theme surface 162:168 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 3 66 theme N-glycosylation 396:410 arg1 sequons 412:418 N-glycosylation sequons 396:418 N-glycosylation sequons 396:418 After a leap to humans from a zoonotic host, influenza can gain N-glycosylation sequons over time as part of its fitness strategy. 26202417 9 67 theme HA 1436:1437 arg1 glycosylation 1439:1451 HA glycosylation 1436:1451 HA glycosylation 1436:1451 Based on this study and previous reports we provide structural insight as to how the immune system responses may differ depending on HA glycosylation. 26202417 1 68 theme glycoprotein 170:181 arg1 target 215:220 the major target 205:220 the major target of host neutralizing antibodies 205:252 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 68 theme glycoprotein 170:181 arg1 HA 198:199 HA 198:199 HA 198:199 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 1 68 theme glycoprotein 170:181 arg1 hemagglutinin 183:195 The influenza virus surface glycoprotein hemagglutinin 142:195 The influenza virus surface glycoprotein hemagglutinin (HA) 142:200 The influenza virus surface glycoprotein hemagglutinin (HA) is the major target of host neutralizing antibodies. 26202417 7 69 dep composition 864:874 arg1 profiling 947:955 permethylation profiling 932:955 permethylation profiling 932:955 We have used: nano-LC-MS(E) for glycopeptide composition, sequence and site occupancy analysis, and MALDI-TOF MS permethylation profiling for characterization of released glycans. 26202417 4 70 theme structural 520:529 arg1 level 531:535 the structural level 516:535 the structural level 516:535 This glycosylation expansion has not been studied at the structural level. 28060516 7 0 theme H5 1462:1463 arg1 structures 1441:1450 the three-dimensional structures 1419:1450 the three-dimensional structures of H3 and H5 1419:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 1 theme insect 593:598 arg1 cells 600:604 (iv) Drosophila S2 insect cells 574:604 (iv) Drosophila S2 insect cells 574:604 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 2 theme glycosylation 1185:1197 arg1 complexity 1171:1180 complexity 1171:1180 complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 1171:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 3 theme H3 1455:1456 arg1 structures 1441:1450 the three-dimensional structures 1419:1450 the three-dimensional structures of H3 and H5 1419:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 4 theme Drosophila 579:588 arg1 cells 600:604 (iv) Drosophila S2 insect cells 574:604 (iv) Drosophila S2 insect cells 574:604 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 5 from N193 1275:1278 arg1 absence 1239:1245 (2) absence 1235:1245 (2) absence of glycosylation at N10 and N193 1235:1278 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 5 from N193 1275:1278 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 3 6 theme glycosylation 722:734 arg1 states 736:741 different glycosylation states 712:741 different glycosylation states 712:741 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 4 7 theme antibody 853:860 arg1 response 862:869 antibody response 853:869 antibody response 853:869 Evidence was found that high-mannose glycans diminished antibody response via DC-SIGN interactions. 28060516 1 8 theme hemagglutinins 214:227 arg1 patterns 182:189 The glycosylation patterns 164:189 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 164:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 3 9 theme immune 750:755 arg1 responses 757:765 the immune responses 746:765 the immune responses in chicken and mouse systems 746:794 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 4 10 theme DC-SIGN 875:881 arg1 interactions 883:894 DC-SIGN interactions 875:894 DC-SIGN interactions 875:894 Evidence was found that high-mannose glycans diminished antibody response via DC-SIGN interactions. 28060516 7 11 theme S2 1381:1382 arg1 cells 1384:1388 HEK293S and S2 cells 1369:1388 cells 1384:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 12 theme canonical 537:545 arg1 glycoform 559:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 13 theme HEK293S 1369:1375 arg1 cells 1384:1388 HEK293S and S2 cells 1369:1388 cells 1384:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 3 14 from effect 702:707 arg1 responses 757:765 the immune responses 746:765 the immune responses in chicken and mouse systems 746:794 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 2 15 with glycoforms 474:483 arg1 glycoform 559:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 5 16 theme MALDI-TOF 950:958 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 17 theme permethylation 963:976 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 7 18 from N10 1267:1269 arg1 absence 1239:1245 (2) absence 1235:1245 (2) absence of glycosylation at N10 and N193 1235:1278 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 18 from N10 1267:1269 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 19 theme mannose 1337:1343 arg1 glycans 1345:1351 high mannose glycans 1332:1351 high mannose glycans 1332:1351 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 20 gly glycoform 559:567 arg1 Man5GlcNAc2 547:557 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 6 21 theme Glycosylation 1069:1081 arg1 occupancy 1088:1096 Glycosylation site occupancy 1069:1096 Glycosylation site occupancy 1069:1096 Glycosylation site occupancy was also determined by LC-MSE. 28060516 2 22 theme GnTI 500:503 arg1 cells 508:512 (iii) HEK293S GnTI(-) cells 486:512 (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform 486:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 23 theme Glycosylation 89:101 arg1 Patterns 103:110 Engineered Glycosylation Patterns 78:110 Engineered Glycosylation Patterns 78:110 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 1 24 theme recombinant 199:209 arg1 hemagglutinins 214:227 four recombinant H5 hemagglutinins 194:227 four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 194:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 5 25 theme LC-MSE 1011:1016 arg1 analysis 1018:1025 LC-MSE analysis 1011:1025 LC-MSE analysis of glycosylation site microheterogeneity 1011:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 1 26 theme glycosylation 168:180 arg1 patterns 182:189 The glycosylation patterns 164:189 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 164:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 2 27 with cells 508:512 arg1 glycoform 559:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 28 dep glycans 1293:1299 arg1 3 1282:1282 3 1282:1282 3 1282:1282 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 29 theme high 1332:1335 arg1 glycans 1345:1351 high mannose glycans 1332:1351 high mannose glycans 1332:1351 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 30 theme HEK293T 392:398 arg1 cells 400:404 HEK293T cells 392:404 HEK293T cells 392:404 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 31 theme Glycosylation 0:12 arg1 Characterization 14:29 Glycosylation Characterization 0:29 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 0:162 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 2 32 theme paucimannose 626:637 arg1 glycoforms 639:648 primarily paucimannose glycoforms 616:648 primarily paucimannose glycoforms 616:648 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 33 from site 1361:1364 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 33 from site 1361:1364 arg1 cells 1384:1388 HEK293S and S2 cells 1369:1388 cells 1384:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 33 from site 1361:1364 arg1 glycans 1293:1299 (3) complex glycans 1281:1299 (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells 1281:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 34 theme globular 1220:1227 arg1 absence 1239:1245 (2) absence 1235:1245 (2) absence of glycosylation at N10 and N193 1235:1278 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 34 theme globular 1220:1227 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 34 theme globular 1220:1227 arg1 glycans 1293:1299 (3) complex glycans 1281:1299 (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells 1281:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 5 35 theme glycans 999:1005 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 35 theme glycans 999:1005 arg1 analysis 1018:1025 LC-MSE analysis 1011:1025 LC-MSE analysis of glycosylation site microheterogeneity 1011:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 7 36 theme HEK293T 1312:1318 arg1 HA 1325:1326 HEK293T cell HA 1312:1326 HEK293T cell HA 1312:1326 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 37 theme Vibrio 419:424 arg1 neuraminidase 434:446 Vibrio cholera neuraminidase 419:446 Vibrio cholera neuraminidase 419:446 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 38 theme Influenza 37:45 arg1 Series 66:71 an Influenza H5N7 Hemagglutinin Series 34:71 an Influenza H5N7 Hemagglutinin Series 34:71 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 7 39 theme glycan 1486:1491 arg1 preferences 1498:1508 glycan type preferences 1486:1508 glycan type preferences 1486:1508 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 40 from complexity 1171:1180 arg1 stem 1208:1211 the stem 1204:1211 the stem 1204:1211 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 41 theme complex 367:373 arg1 glycoforms 375:384 complex glycoforms 367:384 complex glycoforms 367:384 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 42 theme Hemagglutinin 52:64 arg1 Series 66:71 an Influenza H5N7 Hemagglutinin Series 34:71 an Influenza H5N7 Hemagglutinin Series 34:71 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 5 43 theme site 1044:1047 arg1 microheterogeneity 1049:1066 glycosylation site microheterogeneity 1030:1066 glycosylation site microheterogeneity 1030:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 2 44 dep treated 406:412 arg1 ii 388:389 ii 388:389 ii 388:389 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 3 45 dep these 663:667 arg1 HAs 669:671 HAs 669:671 HAs 669:671 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 2 46 theme S2 590:591 arg1 cells 600:604 (iv) Drosophila S2 insect cells 574:604 (iv) Drosophila S2 insect cells 574:604 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 47 theme three-dimensional 1423:1439 arg1 structures 1441:1450 the three-dimensional structures 1419:1450 the three-dimensional structures of H3 and H5 1419:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 3 48 theme states 736:741 arg1 effect 702:707 the effect 698:707 the effect of different glycosylation states on the immune responses in chicken and mouse systems 698:794 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 1 49 theme H5 211:212 arg1 hemagglutinins 214:227 four recombinant H5 hemagglutinins 194:227 four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 194:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 7 50 theme 4 1396:1396 arg1 differences 1399:1409 the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences 1216:1409 the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 1216:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 8 51 theme findings 1560:1567 arg1 implications 1540:1551 Biological implications 1529:1551 Biological implications of the findings 1529:1567 Biological implications of the findings are discussed. 28060516 3 52 theme different 712:720 arg1 states 736:741 different glycosylation states 712:741 different glycosylation states 712:741 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 7 53 theme glycosylation 1250:1262 arg1 absence 1239:1245 (2) absence 1235:1245 (2) absence of glycosylation at N10 and N193 1235:1278 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 53 theme glycosylation 1250:1262 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 54 theme HEK293T 342:348 arg1 cells 350:354 (i) HEK293T cells 338:354 (i) HEK293T cells 338:354 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 2 55 dep cells 600:604 arg1 iv 575:576 iv 575:576 iv 575:576 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 56 theme major 1133:1137 arg1 findings 1139:1146 Our major findings 1129:1146 Our major findings 1129:1146 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 1 57 dep hemagglutinins 214:227 arg1 HAs 230:232 HAs 230:232 HAs 230:232 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 2 58 with cells 600:604 arg1 glycoform 559:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 2 59 theme Man5GlcNAc2 547:557 arg1 glycoform 559:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 predominantly the canonical Man5GlcNAc2 glycoform 519:567 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 60 dep include 1148:1154 arg1 1 1157:1157 1 1157:1157 1 1157:1157 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 60 dep include 1148:1154 arg1 decreasing 1160:1169 decreasing 1160:1169 include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites 1148:1526 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 1 61 gly glycosylation 168:180 arg1 hemagglutinins 214:227 four recombinant H5 hemagglutinins 194:227 four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 194:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 5 62 theme MS 960:961 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 0 63 theme Engineered 78:87 arg1 Patterns 103:110 Engineered Glycosylation Patterns 78:110 Engineered Glycosylation Patterns 78:110 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 6 64 theme site 1083:1086 arg1 occupancy 1088:1096 Glycosylation site occupancy 1069:1096 Glycosylation site occupancy 1069:1096 Glycosylation site occupancy was also determined by LC-MSE. 28060516 5 65 theme semiquantitative 914:929 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 65 theme semiquantitative 914:929 arg1 analyses 931:938 two semiquantitative analyses 910:938 two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity 910:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 65 theme semiquantitative 914:929 arg1 analysis 1018:1025 LC-MSE analysis 1011:1025 LC-MSE analysis of glycosylation site microheterogeneity 1011:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 2 66 theme HEK293S 492:498 arg1 GnTI 500:503 (iii) HEK293S GnTI 486:503 (iii) HEK293S GnTI 486:503 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 67 from stem 1208:1211 arg1 complexity 1171:1180 complexity 1171:1180 complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 1171:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 67 from stem 1208:1211 arg1 glycosylation 1185:1197 glycosylation 1185:1197 glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 1185:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 68 gly glycoforms 474:483 arg1 asialo-complex 459:472 asialo-complex glycoforms 459:483 asialo-complex glycoforms 459:483 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 69 theme selected 1513:1520 arg1 sites 1522:1526 selected sites 1513:1526 selected sites 1513:1526 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 0 70 theme Structure-Function 130:147 arg1 Relationships 149:161 Structure-Function Relationships 130:161 Structure-Function Relationships 130:161 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 3 71 dep chicken 770:776 arg1 systems 788:794 systems 788:794 systems 788:794 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 7 72 theme head 1229:1232 arg1 differences 1399:1409 the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences 1216:1409 the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 1216:1463 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 73 theme cell 1320:1323 arg1 HA 1325:1326 HEK293T cell HA 1312:1326 HEK293T cell HA 1312:1326 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 2 74 theme asialo-complex 459:472 arg1 glycoforms 474:483 asialo-complex glycoforms 459:483 asialo-complex glycoforms 459:483 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 3 75 from responses 757:765 arg1 chicken 770:776 chicken 770:776 chicken 770:776 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 3 75 from responses 757:765 arg1 mouse 782:786 mouse 782:786 mouse 782:786 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 2 76 dep cells 350:354 arg1 i 339:339 i 339:339 i 339:339 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 77 dep Characterization 14:29 arg1 Implications 113:124 Implications 113:124 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 0:162 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 5 78 theme released 990:997 arg1 glycans 999:1005 released glycans 990:1005 released glycans 990:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 8 79 theme Biological 1529:1538 arg1 implications 1540:1551 Biological implications 1529:1551 Biological implications of the findings 1529:1567 Biological implications of the findings are discussed. 28060516 2 80 theme cholera 426:432 arg1 neuraminidase 434:446 Vibrio cholera neuraminidase 419:446 Vibrio cholera neuraminidase 419:446 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 0 81 theme H5N7 47:50 arg1 Series 66:71 an Influenza H5N7 Hemagglutinin Series 34:71 an Influenza H5N7 Hemagglutinin Series 34:71 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 5 82 gly glycosylation 1030:1042 arg2 site 1044:1047 glycosylation site microheterogeneity 1030:1066 glycosylation site microheterogeneity 1030:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 82 gly glycosylation 1030:1042 arg2 microheterogeneity 1049:1066 glycosylation site microheterogeneity 1030:1066 glycosylation site microheterogeneity 1030:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 0 83 theme Series 66:71 arg1 Characterization 14:29 Glycosylation Characterization 0:29 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 0:162 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 2 84 dep GnTI 500:503 arg1 iii 487:489 iii 487:489 iii 487:489 The proteins were expressed in (i) HEK293T cells to produce complex glycoforms, (ii) HEK293T cells treated with Vibrio cholera neuraminidase to provide asialo-complex glycoforms, (iii) HEK293S GnTI(-) cells with predominantly the canonical Man5GlcNAc2 glycoform, and (iv) Drosophila S2 insect cells producing primarily paucimannose glycoforms. 28060516 7 85 theme type 1493:1496 arg1 preferences 1498:1508 glycan type preferences 1486:1508 glycan type preferences 1486:1508 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 86 from N165 1304:1307 arg1 HA 1325:1326 HEK293T cell HA 1312:1326 HEK293T cell HA 1312:1326 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 86 from N165 1304:1307 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 86 from N165 1304:1307 arg1 glycans 1345:1351 high mannose glycans 1332:1351 high mannose glycans 1332:1351 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 86 from N165 1304:1307 arg1 glycans 1293:1299 (3) complex glycans 1281:1299 (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells 1281:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 87 theme complex 1285:1291 arg1 head 1229:1232 the globular head 1216:1232 the globular head 1216:1232 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 7 87 theme complex 1285:1291 arg1 glycans 1293:1299 (3) complex glycans 1281:1299 (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells 1281:1388 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 3 88 used used 678:681 arg2 these 663:667 these 663:667 these 663:667 Previously, these HAs were used to investigate the effect of different glycosylation states on the immune responses in chicken and mouse systems. 28060516 1 89 attach derived 235:241 arg1 H5N7 276:279 H5N7 276:279 H5N7 276:279 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 1 89 attach derived 235:241 arg2 hemagglutinins 214:227 four recombinant H5 hemagglutinins 194:227 four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) 194:280 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 1 89 attach derived 235:241 arg1 A/Mallard/Denmark/64650/03 248:273 A/Mallard/Denmark/64650/03 248:273 A/Mallard/Denmark/64650/03 248:273 The glycosylation patterns of four recombinant H5 hemagglutinins (HAs) derived from A/Mallard/Denmark/64650/03 (H5N7) have been characterized. 28060516 4 90 theme high-mannose 821:832 arg1 glycans 834:840 high-mannose glycans 821:840 high-mannose glycans 821:840 Evidence was found that high-mannose glycans diminished antibody response via DC-SIGN interactions. 28060516 0 91 with Characterization 14:29 arg1 Patterns 103:110 Engineered Glycosylation Patterns 78:110 Engineered Glycosylation Patterns 78:110 Glycosylation Characterization of an Influenza H5N7 Hemagglutinin Series with Engineered Glycosylation Patterns: Implications for Structure-Function Relationships. 28060516 5 92 theme glycosylation 1030:1042 arg1 microheterogeneity 1049:1066 glycosylation site microheterogeneity 1030:1066 glycosylation site microheterogeneity 1030:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 7 93 dep absence 1239:1245 arg1 2 1236:1236 2 1236:1236 2 1236:1236 Our major findings include (1) decreasing complexity of glycosylation from the stem to the globular head, (2) absence of glycosylation at N10 and N193, (3) complex glycans at N165 in HEK293T cell HA but high mannose glycans at this site in HEK293S and S2 cells, and (4) differences between the three-dimensional structures of H3 and H5 HAs that may explain glycan type preferences at selected sites. 28060516 5 94 theme microheterogeneity 1049:1066 arg1 analysis 978:985 MALDI-TOF MS permethylation analysis 950:985 MALDI-TOF MS permethylation analysis of released glycans 950:1005 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 28060516 5 94 theme microheterogeneity 1049:1066 arg1 analysis 1018:1025 LC-MSE analysis 1011:1025 LC-MSE analysis of glycosylation site microheterogeneity 1011:1066 We performed two semiquantitative analyses including MALDI-TOF MS permethylation analysis of released glycans and LC-MSE analysis of glycosylation site microheterogeneity. 25660649 10 0 from asparagine 1432:1441 arg1 glycoforms 1418:1427 Predominant glycoforms 1406:1427 Predominant glycoforms at asparagine 67 1406:1444 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 10 0 from asparagine 1432:1441 arg1 structures 1468:1477 high mannose-type structures 1450:1477 high mannose-type structures 1450:1477 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 12 1 theme CYD 1796:1798 arg1 characterizations 1775:1791 Our E-protein glycan characterizations 1754:1791 Our E-protein glycan characterizations of CYD 1754:1798 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 11 2 link monocyte-derived 1720:1735 arg1 cells 1747:1751 human monocyte-derived dendritic cells 1714:1751 human monocyte-derived dendritic cells 1714:1751 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 13 3 theme virus-host 1984:1993 arg1 interactions 2000:2011 the dengue virus-host cell interactions 1973:2011 the dengue virus-host cell interactions 1973:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 4 4 theme tetravalent 685:695 arg1 CYD 713:715 CYD 713:715 CYD 713:715 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 4 4 theme tetravalent 685:695 arg1 vaccine 704:710 The Sanofi Pasteur tetravalent dengue vaccine 666:710 The Sanofi Pasteur tetravalent dengue vaccine (CYD) 666:716 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 11 5 theme wild 1678:1681 arg1 type 1683:1686 the wild type 1674:1686 the wild type parents 1674:1694 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 7 6 theme N-linked 1097:1104 arg1 glycans 1106:1112 the N-linked glycans 1093:1112 the N-linked glycans 1093:1112 In this paper, we investigated the structure of the N-linked glycans in the four CYD serotypes. 25660649 2 7 theme proteins 296:303 arg1 characterization 263:278 Glycosylation characterization 249:278 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) 249:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 12 8 theme E-protein 1758:1766 arg1 characterizations 1775:1791 Our E-protein glycan characterizations 1754:1791 Our E-protein glycan characterizations of CYD 1754:1798 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 8 9 theme high-mannose 1222:1233 arg1 glycans 1255:1261 high-mannose, hybrid and complex glycans 1222:1261 glycans 1255:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 6 10 gly glycosylation 971:983 arg1 DENV 1000:1003 the DENV 996:1003 the DENV 996:1003 Despite the importance of this interaction, the specific glycosylation pattern of the DENV has not been clearly documented so far. 25660649 2 11 theme cell-derived 334:345 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 11 theme cell-derived 334:345 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 1 12 theme fundamental 196:206 arg1 role 208:211 a fundamental role 194:211 a fundamental role 194:211 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 8 13 theme CYDs 1191:1194 arg1 glycans 1180:1186 the N-linked glycans 1167:1186 the N-linked glycans of CYDs 1167:1194 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 8 13 theme CYDs 1191:1194 arg1 mix 1215:1217 a mix 1213:1217 a mix of high-mannose, hybrid and complex glycans 1213:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 8 14 theme hybrid 1236:1241 arg1 glycans 1255:1261 high-mannose, hybrid and complex glycans 1222:1261 glycans 1255:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 3 15 theme virus 582:586 arg1 production 588:597 virus production 582:597 virus production 582:597 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 5 16 theme dendritic 846:854 arg1 cells 856:860 human monocyte-derived dendritic cells 823:860 human monocyte-derived dendritic cells 823:860 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 6 17 theme glycosylation 971:983 arg1 pattern 985:991 the specific glycosylation pattern 958:991 the specific glycosylation pattern of the DENV 958:1003 Despite the importance of this interaction, the specific glycosylation pattern of the DENV has not been clearly documented so far. 25660649 2 18 theme potential 392:400 arg1 residues 416:423 two potential glycosylation residues 388:423 two potential glycosylation residues 388:423 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 18 theme potential 392:400 arg1 asparagine 430:439 the asparagine 426:439 the asparagine 67 and 153 426:450 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 8 19 theme complex 1247:1253 arg1 glycans 1255:1261 high-mannose, hybrid and complex glycans 1222:1261 glycans 1255:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 1 20 theme virus-host 220:229 arg1 interaction 236:246 the virus-host cell interaction 216:246 the virus-host cell interaction 216:246 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 3 21 theme infection 607:615 arg1 spreading 617:625 the infection spreading 603:625 the infection spreading 603:625 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 1 22 theme several 129:135 arg1 studies 143:149 several virus studies 129:149 several virus studies 129:149 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 6 23 theme interaction 945:955 arg1 importance 926:935 the importance 922:935 the importance of this interaction 922:955 Despite the importance of this interaction, the specific glycosylation pattern of the DENV has not been clearly documented so far. 25660649 10 24 located detected 1532:1539 arg1 asparagine 1544:1553 asparagine 153 1544:1557 asparagine 153 1544:1557 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 10 24 located detected 1532:1539 arg2 structures 1517:1526 complex- and hybrid-type structures 1492:1526 complex- and hybrid-type structures 1492:1526 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 3 25 theme mammalian 649:657 arg1 cells 659:663 mammalian cells 649:663 mammalian cells 649:663 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 9 26 gly N-glycosylation 1278:1292 arg1 CYDs 1306:1309 CYDs 1306:1309 CYDs using nanoLC-ESI-MS/MS 1306:1332 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 10 27 theme mannose-type 1455:1466 arg1 glycoforms 1418:1427 Predominant glycoforms 1406:1427 Predominant glycoforms at asparagine 67 1406:1444 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 10 27 theme mannose-type 1455:1466 arg1 structures 1468:1477 high mannose-type structures 1450:1477 high mannose-type structures 1450:1477 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 12 28 dep consistent 1804:1813 arg1 support 1875:1881 support 1875:1881 support in vitro studies 1875:1898 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 0 29 theme tetravalent 87:97 arg1 vaccine 110:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 9 30 theme Site-specific 1264:1276 arg1 analysis 1294:1301 Site-specific N-glycosylation analysis 1264:1301 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS 1264:1332 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 1 31 theme protein 167:173 arg1 glycosylation 175:187 protein glycosylation 167:187 protein glycosylation 167:187 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 11 32 from comparable 1700:1709 arg1 cells 1747:1751 human monocyte-derived dendritic cells 1714:1751 human monocyte-derived dendritic cells 1714:1751 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 0 33 theme dengue 103:108 arg1 vaccine 110:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 6 34 theme DENV 1000:1003 arg1 pattern 985:991 the specific glycosylation pattern 958:991 the specific glycosylation pattern of the DENV 958:1003 Despite the importance of this interaction, the specific glycosylation pattern of the DENV has not been clearly documented so far. 25660649 10 35 theme complex- 1492:1499 arg1 structures 1517:1526 complex- and hybrid-type structures 1492:1526 complex- and hybrid-type structures 1492:1526 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 9 36 theme CYDs 1306:1309 arg1 analysis 1294:1301 Site-specific N-glycosylation analysis 1264:1301 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS 1264:1332 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 2 37 theme glycosylation 402:414 arg1 residues 416:423 two potential glycosylation residues 388:423 two potential glycosylation residues 388:423 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 37 theme glycosylation 402:414 arg1 asparagine 430:439 the asparagine 426:439 the asparagine 67 and 153 426:450 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 0 38 theme Site-specific 0:12 arg1 characterization 14:29 Site-specific characterization 0:29 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 0:117 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 11 39 dep viruses 1655:1661 arg1 1-4 1663:1665 1-4 1663:1665 1-4 1663:1665 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 13 40 theme cell 1995:1998 arg1 interactions 2000:2011 the dengue virus-host cell interactions 1973:2011 the dengue virus-host cell interactions 1973:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 4 41 theme vero 774:777 arg1 cells 779:783 mammalian vero cells 764:783 mammalian vero cells 764:783 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 5 42 theme human 823:827 arg1 cells 856:860 human monocyte-derived dendritic cells 823:860 human monocyte-derived dendritic cells 823:860 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 10 43 theme hybrid-type 1505:1515 arg1 structures 1517:1526 complex- and hybrid-type structures 1492:1526 complex- and hybrid-type structures 1492:1526 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 11 44 theme monocyte-derived 1720:1735 arg1 cells 1747:1751 human monocyte-derived dendritic cells 1714:1751 human monocyte-derived dendritic cells 1714:1751 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 0 45 theme protein 43:49 arg1 N-glycosylation 51:65 envelope protein N-glycosylation 34:65 envelope protein N-glycosylation 34:65 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 9 46 theme asparagine 1357:1366 arg1 residues 1368:1375 both asparagine residues 67 and 153 1352:1386 both asparagine residues 67 and 153 1352:1386 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 12 47 theme type 1853:1856 arg1 parents 1858:1864 the wild type parents 1844:1864 the wild type parents 1844:1864 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 12 48 with consistent 1804:1813 arg1 observations 1826:1837 those observations 1820:1837 those observations from the wild type parents 1820:1864 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 11 49 theme infection 1627:1635 arg1 comparable 1700:1709 comparable 1700:1709 comparable 1700:1709 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 49 theme infection 1627:1635 arg1 consequences 1611:1622 the immunological consequences 1593:1622 the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents 1593:1694 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 12 50 from parents 1858:1864 arg1 observations 1826:1837 those observations 1820:1837 those observations from the wild type parents 1820:1864 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 1 51 theme cell 231:234 arg1 interaction 236:246 the virus-host cell interaction 216:246 the virus-host cell interaction 216:246 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 12 52 theme in 1883:1884 arg1 studies 1892:1898 in vitro studies 1883:1898 in vitro studies 1883:1898 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 11 53 theme dengue 1648:1653 arg1 viruses 1655:1661 the CYD dengue viruses 1-4 1640:1665 the CYD dengue viruses 1-4 1640:1665 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 7 54 theme CYD 1126:1128 arg1 serotypes 1130:1138 the four CYD serotypes 1117:1138 the four CYD serotypes 1117:1138 In this paper, we investigated the structure of the N-linked glycans in the four CYD serotypes. 25660649 7 55 theme glycans 1106:1112 arg1 structure 1080:1088 the structure 1076:1088 the structure of the N-linked glycans in the four CYD serotypes 1076:1138 In this paper, we investigated the structure of the N-linked glycans in the four CYD serotypes. 25660649 13 56 theme dengue 1977:1982 arg1 interactions 2000:2011 the dengue virus-host cell interactions 1973:2011 the dengue virus-host cell interactions 1973:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 2 57 theme insect 313:318 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 57 theme insect 313:318 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 8 58 theme MALDI-TOF 1147:1155 arg1 analysis 1157:1164 MALDI-TOF analysis 1147:1164 MALDI-TOF analysis 1147:1164 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 11 59 theme In 1560:1561 arg1 studies 1569:1575 In vitro studies 1560:1575 In vitro studies 1560:1575 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 3 60 theme residues 542:549 arg1 glycosylation 515:527 the glycosylation 511:527 the glycosylation of these two residues 511:549 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 12 61 theme glycan 1768:1773 arg1 characterizations 1775:1791 Our E-protein glycan characterizations 1754:1791 Our E-protein glycan characterizations of CYD 1754:1798 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 4 62 theme Pasteur 677:683 arg1 CYD 713:715 CYD 713:715 CYD 713:715 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 4 62 theme Pasteur 677:683 arg1 vaccine 704:710 The Sanofi Pasteur tetravalent dengue vaccine 666:710 The Sanofi Pasteur tetravalent dengue vaccine (CYD) 666:716 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 11 63 theme type 1683:1686 arg1 parents 1688:1694 the wild type parents 1674:1694 the wild type parents 1674:1694 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 2 64 theme dengue 347:352 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 64 theme dengue 347:352 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 5 65 theme C-type 875:880 arg1 lectins 882:888 C-type lectins 875:888 C-type lectins 875:888 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 8 66 theme N-linked 1171:1178 arg1 glycans 1180:1186 the N-linked glycans 1167:1186 the N-linked glycans of CYDs 1167:1194 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 8 66 theme N-linked 1171:1178 arg1 mix 1215:1217 a mix 1213:1217 a mix of high-mannose, hybrid and complex glycans 1213:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 13 67 from role 1954:1957 arg1 interactions 2000:2011 the dengue virus-host cell interactions 1973:2011 the dengue virus-host cell interactions 1973:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 4 68 theme dengue 697:702 arg1 CYD 713:715 CYD 713:715 CYD 713:715 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 4 68 theme dengue 697:702 arg1 vaccine 704:710 The Sanofi Pasteur tetravalent dengue vaccine 666:710 The Sanofi Pasteur tetravalent dengue vaccine (CYD) 666:716 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 13 69 theme glycans 1962:1968 arg1 role 1954:1957 the role 1950:1957 the role of glycans in the dengue virus-host cell interactions 1950:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 10 70 theme Predominant 1406:1416 arg1 glycoforms 1418:1427 Predominant glycoforms 1406:1427 Predominant glycoforms at asparagine 67 1406:1444 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 10 70 theme Predominant 1406:1416 arg1 structures 1468:1477 high mannose-type structures 1450:1477 high mannose-type structures 1450:1477 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 2 71 theme mammalian 324:332 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 71 theme mammalian 324:332 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 5 72 theme monocyte-derived 829:844 arg1 cells 856:860 human monocyte-derived dendritic cells 823:860 human monocyte-derived dendritic cells 823:860 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 2 73 theme Glycosylation 249:261 arg1 characterization 263:278 Glycosylation characterization 249:278 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) 249:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 6 74 theme specific 962:969 arg1 pattern 985:991 the specific glycosylation pattern 958:991 the specific glycosylation pattern of the DENV 958:1003 Despite the importance of this interaction, the specific glycosylation pattern of the DENV has not been clearly documented so far. 25660649 8 75 theme glycans 1255:1261 arg1 mix 1215:1217 a mix 1213:1217 a mix of high-mannose, hybrid and complex glycans 1213:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 8 75 theme glycans 1255:1261 arg1 glycans 1180:1186 the N-linked glycans 1167:1186 the N-linked glycans of CYDs 1167:1194 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 13 76 theme new 1933:1935 arg1 insights 1937:1944 new insights 1933:1944 new insights for the role of glycans in the dengue virus-host cell interactions 1933:2011 In addition, these data provide new insights for the role of glycans in the dengue virus-host cell interactions. 25660649 2 77 link cell-derived 334:345 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 77 link cell-derived 334:345 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 78 theme envelope 287:294 arg1 proteins 296:303 the envelope proteins 283:303 the envelope proteins 283:303 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 1 79 theme virus 137:141 arg1 studies 143:149 several virus studies 129:149 several virus studies 129:149 Recently, several virus studies have shown that protein glycosylation play a fundamental role in the virus-host cell interaction. 25660649 4 80 theme chimeric 735:742 arg1 viruses 744:750 four chimeric viruses 730:750 four chimeric viruses produced in mammalian vero cells 730:783 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 3 81 from production 588:597 arg1 mosquito 637:644 mosquito 637:644 mosquito 637:644 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 3 81 from production 588:597 arg1 cells 659:663 mammalian cells 649:663 mammalian cells 649:663 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 5 82 theme cell-surface 890:901 arg1 CYDs 799:802 the CYDs 795:802 the CYDs 795:802 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 5 82 theme cell-surface 890:901 arg1 molecules 903:911 able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules 808:911 able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules 808:911 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 11 83 from cells 1747:1751 arg1 comparable 1700:1709 comparable 1700:1709 comparable 1700:1709 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 83 from cells 1747:1751 arg1 consequences 1611:1622 the immunological consequences 1593:1622 the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents 1593:1694 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 0 84 theme CYD 99:101 arg1 vaccine 110:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 3 85 dep production 588:597 arg1 the 578:580 the 578:580 the 578:580 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 10 86 theme high 1450:1453 arg1 glycoforms 1418:1427 Predominant glycoforms 1406:1427 Predominant glycoforms at asparagine 67 1406:1444 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 10 86 theme high 1450:1453 arg1 structures 1468:1477 high mannose-type structures 1450:1477 high mannose-type structures 1450:1477 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 9 87 theme N-glycosylation 1278:1292 arg1 analysis 1294:1301 Site-specific N-glycosylation analysis 1264:1301 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS 1264:1332 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 5 88 link monocyte-derived 829:844 arg1 cells 856:860 human monocyte-derived dendritic cells 823:860 human monocyte-derived dendritic cells 823:860 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 8 89 link N-linked 1171:1178 arg1 glycans 1180:1186 the N-linked glycans 1167:1186 the N-linked glycans of CYDs 1167:1194 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 8 89 link N-linked 1171:1178 arg1 mix 1215:1217 a mix 1213:1217 a mix of high-mannose, hybrid and complex glycans 1213:1261 Using MALDI-TOF analysis, the N-linked glycans of CYDs were found to be a mix of high-mannose, hybrid and complex glycans. 25660649 3 90 gly glycosylation 515:527 arg1 residues 542:549 these two residues 532:549 these two residues 532:549 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 5 91 theme able 808:811 arg1 CYDs 799:802 the CYDs 795:802 the CYDs 795:802 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 5 91 theme able 808:811 arg1 molecules 903:911 able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules 808:911 able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules 808:911 As DENV, the CYDs are able to infect human monocyte-derived dendritic cells in vitro via C-type lectins cell-surface molecules. 25660649 7 92 from structure 1080:1088 arg1 serotypes 1130:1138 the four CYD serotypes 1117:1138 the four CYD serotypes 1117:1138 In this paper, we investigated the structure of the N-linked glycans in the four CYD serotypes. 25660649 7 93 link N-linked 1097:1104 arg1 glycans 1106:1112 the N-linked glycans 1093:1112 the N-linked glycans 1093:1112 In this paper, we investigated the structure of the N-linked glycans in the four CYD serotypes. 25660649 0 94 theme envelope 34:41 arg1 N-glycosylation 51:65 envelope protein N-glycosylation 34:65 envelope protein N-glycosylation 34:65 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 2 95 from characterization 263:278 arg1 DENV 361:364 DENV 361:364 DENV 361:364 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 95 from characterization 263:278 arg1 virus 354:358 both insect and mammalian cell-derived dengue virus 308:358 both insect and mammalian cell-derived dengue virus (DENV) 308:365 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 10 96 gly glycoforms 1418:1427 arg2 asparagine 1432:1441 asparagine 67 1432:1444 asparagine 67 1432:1444 Predominant glycoforms at asparagine 67 are high mannose-type structures while mainly complex- and hybrid-type structures are detected at asparagine 153. 25660649 11 97 theme human 1714:1718 arg1 cells 1747:1751 human monocyte-derived dendritic cells 1714:1751 human monocyte-derived dendritic cells 1714:1751 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 4 98 theme mammalian 764:772 arg1 cells 779:783 mammalian vero cells 764:783 mammalian vero cells 764:783 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 3 99 from spreading 617:625 arg1 mosquito 637:644 mosquito 637:644 mosquito 637:644 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 3 99 from spreading 617:625 arg1 cells 659:663 mammalian cells 649:663 mammalian cells 649:663 Moreover, it appears that the glycosylation of these two residues can influence dramatically the virus production and the infection spreading in either mosquito or mammalian cells. 25660649 0 100 theme N-glycosylation 51:65 arg1 characterization 14:29 Site-specific characterization 0:29 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 0:117 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 4 101 theme Sanofi 670:675 arg1 CYD 713:715 CYD 713:715 CYD 713:715 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 4 101 theme Sanofi 670:675 arg1 vaccine 704:710 The Sanofi Pasteur tetravalent dengue vaccine 666:710 The Sanofi Pasteur tetravalent dengue vaccine (CYD) 666:716 The Sanofi Pasteur tetravalent dengue vaccine (CYD) consists of four chimeric viruses produced in mammalian vero cells. 25660649 0 102 theme Sanofi 70:75 arg1 vaccine 110:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Sanofi Pasteur's tetravalent CYD dengue vaccine 70:116 Site-specific characterization of envelope protein N-glycosylation on Sanofi Pasteur's tetravalent CYD dengue vaccine. 25660649 2 103 gly glycosylated 471:482 arg1 residues 416:423 two potential glycosylation residues 388:423 two potential glycosylation residues 388:423 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 2 103 gly glycosylated 471:482 arg1 asparagine 430:439 the asparagine 426:439 the asparagine 67 and 153 426:450 Glycosylation characterization of the envelope proteins in both insect and mammalian cell-derived dengue virus (DENV) has established that two potential glycosylation residues, the asparagine 67 and 153 can potentially be glycosylated. 25660649 12 104 theme wild 1848:1851 arg1 parents 1858:1864 the wild type parents 1844:1864 the wild type parents 1844:1864 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 25660649 9 105 gly glycosylated 1392:1403 arg1 residues 1368:1375 both asparagine residues 67 and 153 1352:1386 both asparagine residues 67 and 153 1352:1386 Site-specific N-glycosylation analysis of CYDs using nanoLC-ESI-MS/MS demonstrates that both asparagine residues 67 and 153 are glycosylated. 25660649 11 106 theme immunological 1597:1609 arg1 comparable 1700:1709 comparable 1700:1709 comparable 1700:1709 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 106 theme immunological 1597:1609 arg1 consequences 1611:1622 the immunological consequences 1593:1622 the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents 1593:1694 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 107 dep In 1560:1561 arg1 vitro 1563:1567 vitro 1563:1567 vitro 1563:1567 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 108 theme dendritic 1737:1745 arg1 cells 1747:1751 human monocyte-derived dendritic cells 1714:1751 human monocyte-derived dendritic cells 1714:1751 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 11 109 theme CYD 1644:1646 arg1 viruses 1655:1661 the CYD dengue viruses 1-4 1640:1665 the CYD dengue viruses 1-4 1640:1665 In vitro studies have shown that the immunological consequences of infection by the CYD dengue viruses 1-4 versus the wild type parents are comparable in human monocyte-derived dendritic cells. 25660649 12 110 dep in 1883:1884 arg1 vitro 1886:1890 vitro 1886:1890 vitro 1886:1890 Our E-protein glycan characterizations of CYD are consistent with those observations from the wild type parents and thus support in vitro studies. 26812091 5 0 theme complementary 727:739 arg1 methods 752:758 Classic and complementary analytical methods 715:758 methods 752:758 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 0 theme complementary 727:739 arg1 bioassay 827:834 the Steelman-Pohley bioassay 807:834 the Steelman-Pohley bioassay 807:834 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 0 theme complementary 727:739 arg1 focusing 793:800 focusing 793:800 focusing 793:800 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 0 theme complementary 727:739 arg1 SDS-PAGE 771:778 SDS-PAGE 771:778 SDS-PAGE 771:778 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 4 1 gly N-glycosylation 609:623 arg2 masses 590:595 Intact and subunit masses 571:595 masses 590:595 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 1 gly N-glycosylation 609:623 arg2 sites 625:629 N-glycosylation sites 609:629 N-glycosylation sites 609:629 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 9 2 theme glycan 1383:1388 arg1 structures 1390:1399 the critical glycan structures 1370:1399 the critical glycan structures of hFSH 1370:1407 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 1 3 theme important 141:149 arg1 Glycosylation 121:133 Glycosylation 121:133 Glycosylation 121:133 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 1 3 theme important 141:149 arg1 PTM 151:153 an important PTM 138:153 an important PTM 138:153 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 9 4 theme hFSH 1404:1407 arg1 structures 1390:1399 the critical glycan structures 1370:1399 the critical glycan structures of hFSH 1370:1407 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 5 5 theme analytical 741:750 arg1 methods 752:758 Classic and complementary analytical methods 715:758 methods 752:758 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 5 theme analytical 741:750 arg1 bioassay 827:834 the Steelman-Pohley bioassay 807:834 the Steelman-Pohley bioassay 807:834 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 5 theme analytical 741:750 arg1 focusing 793:800 focusing 793:800 focusing 793:800 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 5 theme analytical 741:750 arg1 SDS-PAGE 771:778 SDS-PAGE 771:778 SDS-PAGE 771:778 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 7 6 theme hFSHs 1108:1112 arg1 O-glycopeptides 1084:1098 The O-glycopeptides 1080:1098 The O-glycopeptides 1080:1098 The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. 26812091 3 7 theme recombinant 492:502 arg1 rhFSH 515:519 rhFSH 515:519 rhFSH 515:519 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 7 theme recombinant 492:502 arg1 FSH 510:512 a recombinant human FSH 490:512 a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells 490:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 4 8 theme mass 696:699 arg1 spectrometry 701:712 mass spectrometry 696:712 mass spectrometry 696:712 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 0 9 theme FSH 89:91 arg1 Assessment 12:21 Comparative Assessment 0:21 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH 0:91 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 6 10 attach predominant 1025:1035 arg2 glycans 1013:1019 macro-heterogeneity glycans 993:1019 macro-heterogeneity glycans 993:1019 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 6 10 attach predominant 1025:1035 arg2 branched 979:986 branched 979:986 branched 979:986 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 6 10 attach predominant 1025:1035 arg1 uhFSH 1044:1048 the uhFSH 1040:1048 the uhFSH 1040:1048 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 6 10 attach predominant 1025:1035 arg2 sialylated 967:976 sialylated 967:976 sialylated 967:976 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 8 11 located observed 1219:1226 arg1 N-glycopeptides 1235:1249 the N-glycopeptides 1231:1249 the N-glycopeptides of both hFSHs 1231:1263 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 8 11 located observed 1219:1226 arg2 degree 1191:1196 A high degree 1184:1196 A high degree of heterogeneity 1184:1213 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 6 12 from uhFSH 1044:1048 arg1 predominant 1025:1035 predominant 1025:1035 predominant 1025:1035 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 2 13 theme human 295:299 arg1 hormone 322:328 human follicle-stimulating hormone 295:328 human follicle-stimulating hormone (hFSH) 295:335 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 2 13 theme human 295:299 arg1 hFSH 331:334 hFSH 331:334 hFSH 331:334 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 3 14 theme Chinese 536:542 arg1 cells 564:568 Chinese hamster ovary (CHO) cells 536:568 Chinese hamster ovary (CHO) cells 536:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 0 15 theme Purified 80:87 arg1 FSH 89:91 a Highly Purified FSH 71:91 a Highly Purified FSH 71:91 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 5 16 theme intact 872:877 arg1 weights 889:895 their intact molecular weights 866:895 their intact molecular weights 866:895 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 3 17 theme glycoproteomic 346:359 arg1 strategy 361:368 a glycoproteomic strategy 344:368 a glycoproteomic strategy 344:368 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 18 gly glycosylation 395:407 arg1 uhFSH 444:448 uhFSH 444:448 uhFSH 444:448 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 18 gly glycosylation 395:407 arg1 FSH 439:441 a putative highly purified FSH 412:441 a putative highly purified FSH (uhFSH) obtained from human urine 412:475 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 1 19 gly glycoproteins 219:231 arg1 glycoproteins 219:231 therapeutic glycoproteins 207:231 therapeutic glycoproteins 207:231 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 5 20 theme molecular 879:887 arg1 weights 889:895 their intact molecular weights 866:895 their intact molecular weights 866:895 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 8 21 theme high 1186:1189 arg1 degree 1191:1196 A high degree 1184:1196 A high degree of heterogeneity 1184:1213 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 6 22 from predominant 1025:1035 arg1 uhFSH 1044:1048 the uhFSH 1040:1048 the uhFSH 1040:1048 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 2 23 theme biological 270:279 arg1 properties 281:290 the biological properties 266:290 the biological properties of human follicle-stimulating hormone (hFSH) 266:335 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 4 24 theme N-glycosylation 609:623 arg1 sites 625:629 N-glycosylation sites 609:629 N-glycosylation sites 609:629 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 24 theme N-glycosylation 609:623 arg1 masses 590:595 Intact and subunit masses 571:595 masses 590:595 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 9 25 theme useful 1307:1312 arg1 information 1314:1324 useful information 1307:1324 useful information in elucidating and in further investigation the critical glycan structures of hFSH 1307:1407 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 3 26 theme hamster 544:550 arg1 cells 564:568 Chinese hamster ovary (CHO) cells 536:568 Chinese hamster ovary (CHO) cells 536:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 27 theme purified 430:437 arg1 uhFSH 444:448 uhFSH 444:448 uhFSH 444:448 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 27 theme purified 430:437 arg1 FSH 439:441 a putative highly purified FSH 412:441 a putative highly purified FSH (uhFSH) obtained from human urine 412:475 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 0 28 theme Comparative 0:10 arg1 Assessment 12:21 Comparative Assessment 0:21 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH 0:91 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 3 29 theme ovary 552:556 arg1 cells 564:568 Chinese hamster ovary (CHO) cells 536:568 Chinese hamster ovary (CHO) cells 536:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 30 theme FSH 439:441 arg1 glycosylation 395:407 the glycosylation 391:407 the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine 391:475 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 0 31 theme Glycosylation 26:38 arg1 Assessment 12:21 Comparative Assessment 0:21 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH 0:91 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 2 32 theme hormone 322:328 arg1 properties 281:290 the biological properties 266:290 the biological properties of human follicle-stimulating hormone (hFSH) 266:335 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 0 33 theme Human 108:112 arg1 Urine 114:118 Human Urine 108:118 Human Urine 108:118 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 5 34 dep focusing 793:800 arg1 isoelectric 781:791 isoelectric 781:791 isoelectric 781:791 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 2 35 theme follicle-stimulating 301:320 arg1 hormone 322:328 human follicle-stimulating hormone 295:328 human follicle-stimulating hormone (hFSH) 295:335 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 2 35 theme follicle-stimulating 301:320 arg1 hFSH 331:334 hFSH 331:334 hFSH 331:334 Glycan significantly influences the biological properties of human follicle-stimulating hormone (hFSH). 26812091 3 36 theme human 465:469 arg1 urine 471:475 human urine 465:475 human urine 465:475 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 37 theme putative 414:421 arg1 uhFSH 444:448 uhFSH 444:448 uhFSH 444:448 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 37 theme putative 414:421 arg1 FSH 439:441 a putative highly purified FSH 412:441 a putative highly purified FSH (uhFSH) obtained from human urine 412:475 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 5 38 theme Steelman-Pohley 811:825 arg1 bioassay 827:834 the Steelman-Pohley bioassay 807:834 the Steelman-Pohley bioassay 807:834 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 9 39 theme critical 1374:1381 arg1 structures 1390:1399 the critical glycan structures 1370:1399 the critical glycan structures of hFSH 1370:1407 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 9 40 from differences 1270:1280 arg1 glycosylation 1285:1297 glycosylation 1285:1297 glycosylation 1285:1297 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 4 41 theme subunit 582:588 arg1 N-glycans 598:606 N-glycans 598:606 N-glycans 598:606 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 41 theme subunit 582:588 arg1 sites 625:629 N-glycosylation sites 609:629 N-glycosylation sites 609:629 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 41 theme subunit 582:588 arg1 masses 590:595 Intact and subunit masses 571:595 masses 590:595 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 41 theme subunit 582:588 arg1 N- 643:644 intact N- 636:644 intact N- 636:644 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 7 42 gly O-glycopeptides 1084:1098 arg2 O-glycopeptides 1084:1098 The O-glycopeptides 1080:1098 The O-glycopeptides 1080:1098 The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. 26812091 7 42 gly O-glycopeptides 1084:1098 arg1 hFSHs 1108:1112 both hFSHs 1103:1112 both hFSHs 1103:1112 The O-glycopeptides of both hFSHs, which have not been described previously, were characterized herein. 26812091 0 43 theme Human 57:61 arg1 FSH 63:65 a Recombinant Human FSH 43:65 a Recombinant Human FSH 43:65 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 9 44 theme further 1348:1354 arg1 investigation 1356:1368 further investigation 1348:1368 further investigation 1348:1368 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 9 45 from structures 1390:1399 arg1 investigation 1356:1368 further investigation 1348:1368 further investigation 1348:1368 The differences in glycosylation provide useful information in elucidating and in further investigation the critical glycan structures of hFSH. 26812091 0 46 theme Recombinant 45:55 arg1 FSH 63:65 a Recombinant Human FSH 43:65 a Recombinant Human FSH 43:65 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 3 47 theme CHO 559:561 arg1 cells 564:568 Chinese hamster ovary (CHO) cells 536:568 Chinese hamster ovary (CHO) cells 536:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 8 48 theme hFSHs 1259:1263 arg1 N-glycopeptides 1235:1249 the N-glycopeptides 1231:1249 the N-glycopeptides of both hFSHs 1231:1263 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 8 49 gly N-glycopeptides 1235:1249 arg2 N-glycopeptides 1235:1249 the N-glycopeptides 1231:1249 the N-glycopeptides of both hFSHs 1231:1263 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 8 49 gly N-glycopeptides 1235:1249 arg1 hFSHs 1259:1263 both hFSHs 1254:1263 both hFSHs 1254:1263 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 6 50 theme macro-heterogeneity 993:1011 arg1 glycans 1013:1019 macro-heterogeneity glycans 993:1019 macro-heterogeneity glycans 993:1019 Results showed that highly sialylated, branched, and macro-heterogeneity glycans are predominant in the uhFSH compared with those in rhFSH. 26812091 5 51 theme charge 898:903 arg1 variants 905:912 charge variants 898:912 charge variants 898:912 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 52 theme specific 919:926 arg1 activities 928:937 specific activities 919:937 specific activities 919:937 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 4 53 theme Intact 571:576 arg1 N-glycans 598:606 N-glycans 598:606 N-glycans 598:606 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 53 theme Intact 571:576 arg1 sites 625:629 N-glycosylation sites 609:629 N-glycosylation sites 609:629 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 53 theme Intact 571:576 arg1 masses 590:595 Intact and subunit masses 571:595 masses 590:595 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 53 theme Intact 571:576 arg1 N- 643:644 intact N- 636:644 intact N- 636:644 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 54 gly O-glycopeptides 650:664 arg2 O-glycopeptides 650:664 O-glycopeptides 650:664 O-glycopeptides 650:664 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 1 55 theme therapeutic 207:217 arg1 glycoproteins 219:231 therapeutic glycoproteins 207:231 therapeutic glycoproteins 207:231 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 5 56 theme Classic 715:721 arg1 methods 752:758 Classic and complementary analytical methods 715:758 methods 752:758 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 56 theme Classic 715:721 arg1 bioassay 827:834 the Steelman-Pohley bioassay 807:834 the Steelman-Pohley bioassay 807:834 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 56 theme Classic 715:721 arg1 focusing 793:800 focusing 793:800 focusing 793:800 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 5 56 theme Classic 715:721 arg1 SDS-PAGE 771:778 SDS-PAGE 771:778 SDS-PAGE 771:778 Classic and complementary analytical methods, including SDS-PAGE, isoelectric focusing, and the Steelman-Pohley bioassay were also employed to compare their intact molecular weights, charge variants, and specific activities. 26812091 3 57 theme human 504:508 arg1 rhFSH 515:519 rhFSH 515:519 rhFSH 515:519 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 3 57 theme human 504:508 arg1 FSH 510:512 a recombinant human FSH 490:512 a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells 490:568 Using a glycoproteomic strategy, this study compared the glycosylation of a putative highly purified FSH (uhFSH) obtained from human urine with that of a recombinant human FSH (rhFSH) obtained from Chinese hamster ovary (CHO) cells. 26812091 8 58 theme heterogeneity 1201:1213 arg1 degree 1191:1196 A high degree 1184:1196 A high degree of heterogeneity 1184:1213 A high degree of heterogeneity was observed in the N-glycopeptides of both hFSHs. 26812091 1 59 theme glycoproteins 219:231 arg1 manufacture 179:189 manufacture 179:189 manufacture 179:189 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 1 59 theme glycoproteins 219:231 arg1 efficacy 195:202 efficacy 195:202 efficacy 195:202 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 0 60 theme FSH 63:65 arg1 Assessment 12:21 Comparative Assessment 0:21 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH 0:91 Comparative Assessment of Glycosylation of a Recombinant Human FSH and a Highly Purified FSH Extracted from Human Urine. 26812091 1 61 dep manufacture 179:189 arg1 the 175:177 the 175:177 the 175:177 Glycosylation is an important PTM and is critical for the manufacture and efficacy of therapeutic glycoproteins. 26812091 4 62 theme intact 636:641 arg1 masses 590:595 Intact and subunit masses 571:595 masses 590:595 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 26812091 4 62 theme intact 636:641 arg1 N- 643:644 intact N- 636:644 intact N- 636:644 Intact and subunit masses, N-glycans, N-glycosylation sites, and intact N- and O-glycopeptides were analyzed and compared by mass spectrometry. 28167607 0 0 theme Important 73:81 arg1 Sites 99:103 Important N-Glycosylation Sites 73:103 Important N-Glycosylation Sites on Integrin α5 for Cell Migration 73:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 5 1 theme migration 590:598 arg1 ability 600:606 the migration ability 586:606 the migration ability of S3-5 cells 586:620 We found that the migration ability of S3-5 cells was decreased in comparison with that of the WT. 28167607 7 2 theme mechanistic 817:827 arg1 manner 829:834 a mechanistic manner 815:834 a mechanistic manner 815:834 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 4 3 theme integrin 472:479 arg1 α5 481:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 4 4 theme integrin 413:420 arg1 α5 422:423 integrin α5 413:423 integrin α5 413:423 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 6 5 theme fiber 754:758 arg1 formation 760:768 actin stress fiber formation 741:768 actin stress fiber formation 741:768 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 10 6 theme molecular 1353:1361 arg1 N-glycosylation 1296:1310 N-glycosylation 1296:1310 N-glycosylation on the β-propeller domain 1296:1336 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 10 6 theme molecular 1353:1361 arg1 switch 1363:1368 a molecular switch 1351:1368 a molecular switch 1351:1368 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 7 7 theme cell 1017:1020 arg1 surface 1022:1028 the cell surface 1013:1028 the cell surface 1013:1028 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 0 8 theme Sites 99:103 arg1 Characterization 53:68 Characterization 53:68 Characterization 53:68 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 8 theme Sites 99:103 arg1 Identification 34:47 Identification 34:47 Identification 34:47 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 6 9 theme actin 741:745 arg1 formation 760:768 actin stress fiber formation 741:768 actin stress fiber formation 741:768 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 1 10 theme cell 252:255 arg1 adhesion 257:264 cell adhesion 252:264 cell adhesion 252:264 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 0 11 theme N-Glycosylation 83:97 arg1 Sites 99:103 Important N-Glycosylation Sites 73:103 Important N-Glycosylation Sites on Integrin α5 for Cell Migration 73:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 12 from Identification 34:47 arg1 α5 117:118 Integrin α5 108:118 Integrin α5 for Cell Migration 108:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 5 13 theme S3-5 611:614 arg1 cells 616:620 S3-5 cells 611:620 S3-5 cells 611:620 We found that the migration ability of S3-5 cells was decreased in comparison with that of the WT. 28167607 4 14 theme Wild-type 398:406 arg1 α5 481:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 4 15 contain contains 491:498 arg1 α5 481:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 4 15 contain contains 491:498 arg2 N-glycans 506:514 fewer N-glycans 500:514 fewer N-glycans 500:514 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 10 16 from N-glycosylation 1296:1310 arg1 domain 1331:1336 the β-propeller domain 1315:1336 the β-propeller domain 1315:1336 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 7 17 from expression 975:984 arg1 surface 1022:1028 the cell surface 1013:1028 the cell surface 1013:1028 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 0 18 theme Integrin 108:115 arg1 α5 117:118 Integrin α5 108:118 Integrin α5 for Cell Migration 108:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 4 19 theme S3-5 452:455 arg1 α5 481:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 7 20 theme integrin 996:1003 arg1 α5β1 1005:1008 active integrin α5β1 989:1008 active integrin α5β1 989:1008 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 7 21 theme integrin 881:888 arg1 α5β1 890:893 active but not total integrin α5β1 860:893 active but not total integrin α5β1 860:893 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 1 22 theme integrin 163:170 arg1 α5β1 172:175 integrin α5β1 163:175 integrin α5β1 163:175 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 8 23 theme cell 1122:1125 arg1 migration 1127:1135 the cell migration ability, active α5β1 expression, and internalization 1118:1188 migration 1127:1135 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 10 24 gly N-glycosylation 1296:1310 arg1 domain 1331:1336 the β-propeller domain 1315:1336 the β-propeller domain 1315:1336 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 7 25 theme α5β1 1005:1008 arg1 expression 975:984 the enhanced expression 962:984 the enhanced expression of active integrin α5β1 on the cell surface 962:1028 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 6 26 theme kinase 730:735 arg1 levels 690:695 the levels 686:695 the levels of phosphorylated focal adhesion kinase and actin stress fiber formation 686:768 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 1 27 theme α5β1 172:175 arg1 N-glycosylation 144:158 The N-glycosylation 140:158 The N-glycosylation of integrin α5β1 140:175 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 0 28 theme Key 2:4 arg1 Regulator 6:14 A Key Regulator 0:14 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 0:138 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 6 29 theme adhesion 721:728 arg1 kinase 730:735 phosphorylated focal adhesion kinase 700:735 phosphorylated focal adhesion kinase 700:735 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 8 30 theme α5β1 1153:1156 arg1 expression 1158:1167 active α5β1 expression 1146:1167 active α5β1 expression 1146:1167 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 8 31 theme β-propeller 1082:1092 arg1 α5 1104:1105 α5 1104:1105 α5 1104:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 8 31 theme β-propeller 1082:1092 arg1 domain 1094:1099 the β-propeller domain 1078:1099 the β-propeller domain of α5 1078:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 9 32 theme α5-syndecan-4 1234:1246 arg1 formation 1256:1264 α5-syndecan-4 complex formation 1234:1264 α5-syndecan-4 complex formation 1234:1264 Moreover, these N-glycans are critical for α5-syndecan-4 complex formation. 28167607 7 33 theme active 989:994 arg1 α5β1 1005:1008 active integrin α5β1 989:1008 active integrin α5β1 989:1008 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 6 34 theme focal 715:719 arg1 kinase 730:735 phosphorylated focal adhesion kinase 700:735 phosphorylated focal adhesion kinase 700:735 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 10 35 theme α5β1 1397:1400 arg1 dynamics 1385:1392 the dynamics 1381:1392 the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration 1381:1485 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 9 36 theme complex 1248:1254 arg1 formation 1256:1264 α5-syndecan-4 complex formation 1234:1264 α5-syndecan-4 complex formation 1234:1264 Moreover, these N-glycans are critical for α5-syndecan-4 complex formation. 28167607 6 37 theme S3-5 799:802 arg1 mutant 804:809 the S3-5 mutant 795:809 the S3-5 mutant 795:809 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 8 38 theme α5 1104:1105 arg1 α5 1104:1105 α5 1104:1105 α5 1104:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 8 38 theme α5 1104:1105 arg1 domain 1094:1099 the β-propeller domain 1078:1099 the β-propeller domain of α5 1078:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 4 39 theme fewer 500:504 arg1 N-glycans 506:514 fewer N-glycans 500:514 fewer N-glycans 500:514 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 7 40 theme enhanced 966:973 arg1 expression 975:984 the enhanced expression 962:984 the enhanced expression of active integrin α5β1 on the cell surface 962:1028 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 6 41 theme phosphorylated 700:713 arg1 kinase 730:735 phosphorylated focal adhesion kinase 700:735 phosphorylated focal adhesion kinase 700:735 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 7 42 theme α5β1 890:893 arg1 internalization 841:855 the internalization 837:855 the internalization of active but not total integrin α5β1 837:893 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 10 43 theme cell 1409:1412 arg1 surface 1414:1420 the cell surface 1405:1420 the cell surface that in turn is required for optimum adhesion for cell migration 1405:1485 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 8 44 from restoration 1044:1054 arg1 α5 1104:1105 α5 1104:1105 α5 1104:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 8 44 from restoration 1044:1054 arg1 domain 1094:1099 the β-propeller domain 1078:1099 the β-propeller domain of α5 1078:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 10 45 theme optimum 1451:1457 arg1 adhesion 1459:1466 optimum adhesion 1451:1466 optimum adhesion for cell migration 1451:1485 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 0 46 theme Adhesion 24:31 arg1 Regulator 6:14 A Key Regulator 0:14 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 0:138 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 10 47 theme β-propeller 1319:1329 arg1 domain 1331:1336 the β-propeller domain 1315:1336 the β-propeller domain 1315:1336 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 4 48 theme α5 422:423 arg1 α5 481:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5 398:482 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 8 49 theme active 1146:1151 arg1 expression 1158:1167 active α5β1 expression 1146:1167 active α5β1 expression 1146:1167 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 1 50 theme many 199:202 arg1 aspects 216:222 many fundamental aspects 199:222 many fundamental aspects 199:222 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 50 theme many 199:202 arg1 migration 270:278 migration 270:278 migration 270:278 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 50 theme many 199:202 arg1 adhesion 257:264 cell adhesion 252:264 cell adhesion 252:264 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 0 51 theme Cell 19:22 arg1 Adhesion 24:31 Cell Adhesion 19:31 Cell Adhesion 19:31 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 10 52 from dynamics 1385:1392 arg1 surface 1414:1420 the cell surface 1405:1420 the cell surface that in turn is required for optimum adhesion for cell migration 1405:1485 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 0 53 theme Cell 124:127 arg1 Migration 129:137 Cell Migration 124:137 Cell Migration 124:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 54 from Characterization 53:68 arg1 α5 117:118 Integrin α5 108:118 Integrin α5 for Cell Migration 108:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 4 55 dep sites 458:462 arg1 5 469:469 5 469:469 5 469:469 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 1 56 theme fundamental 204:214 arg1 aspects 216:222 many fundamental aspects 199:222 many fundamental aspects 199:222 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 56 theme fundamental 204:214 arg1 migration 270:278 migration 270:278 migration 270:278 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 56 theme fundamental 204:214 arg1 adhesion 257:264 cell adhesion 252:264 cell adhesion 252:264 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 3 57 theme loss-of-function 371:386 arg1 approach 388:395 a loss-of-function approach 369:395 a loss-of-function approach 369:395 Here, we used a loss-of-function approach. 28167607 8 58 theme N-glycosylation 1059:1073 arg1 restoration 1044:1054 restoration 1044:1054 restoration of N-glycosylation on the β-propeller domain of α5 1044:1105 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 5 59 theme cells 616:620 arg1 ability 600:606 the migration ability 586:606 the migration ability of S3-5 cells 586:620 We found that the migration ability of S3-5 cells was decreased in comparison with that of the WT. 28167607 4 60 theme N-glycosylation 429:443 arg1 sites 458:462 sites 3 to 5 458:469 sites 3 to 5 458:469 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 4 60 theme N-glycosylation 429:443 arg1 S3-5 452:455 N-glycosylation mutant S3-5 429:455 N-glycosylation mutant S3-5 (sites 3 to 5) 429:470 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 7 61 theme total 875:879 arg1 α5β1 890:893 active but not total integrin α5β1 860:893 active but not total integrin α5β1 860:893 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 3 62 used used 364:367 arg2 we 361:362 we 361:362 we 361:362 Here, we used a loss-of-function approach. 28167607 4 63 dep 5 469:469 arg1 to 466:467 to 466:467 to 466:467 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 8 64 dep migration 1127:1135 arg1 ability 1137:1143 ability 1137:1143 ability 1137:1143 Importantly, restoration of N-glycosylation on the β-propeller domain of α5 reinstated the cell migration ability, active α5β1 expression, and internalization. 28167607 4 65 theme mutant 445:450 arg1 sites 458:462 sites 3 to 5 458:469 sites 3 to 5 458:469 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 4 65 theme mutant 445:450 arg1 S3-5 452:455 N-glycosylation mutant S3-5 429:455 N-glycosylation mutant S3-5 (sites 3 to 5) 429:470 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 7 66 theme active 860:865 arg1 α5β1 890:893 active but not total integrin α5β1 860:893 active but not total integrin α5β1 860:893 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 6 67 theme stress 747:752 arg1 formation 760:768 actin stress fiber formation 741:768 actin stress fiber formation 741:768 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 4 68 theme cancer 558:563 arg1 cells 565:569 α5 knockout cancer cells 546:569 α5 knockout cancer cells 546:569 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 1 69 gly N-glycosylation 144:158 arg1 α5β1 172:175 integrin α5β1 163:175 integrin α5β1 163:175 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 0 70 from Sites 99:103 arg1 α5 117:118 Integrin α5 108:118 Integrin α5 for Cell Migration 108:137 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 71 dep Regulator 6:14 arg1 Characterization 53:68 Characterization 53:68 Characterization 53:68 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 71 dep Regulator 6:14 arg1 Identification 34:47 Identification 34:47 Identification 34:47 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 7 72 theme S3-5 912:915 arg1 process 935:941 a process 933:941 a process that is related to the enhanced expression of active integrin α5β1 on the cell surface 933:1028 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 7 72 theme S3-5 912:915 arg1 cells 917:921 S3-5 cells 912:921 S3-5 cells 912:921 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 7 72 theme S3-5 912:915 arg1 related 951:957 related 951:957 related 951:957 In a mechanistic manner, the internalization of active but not total integrin α5β1 was inhibited in S3-5 cells, which is a process that is related to the enhanced expression of active integrin α5β1 on the cell surface. 28167607 4 73 theme knockout 549:556 arg1 cells 565:569 α5 knockout cancer cells 546:569 α5 knockout cancer cells 546:569 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 6 74 theme formation 760:768 arg1 levels 690:695 the levels 686:695 the levels of phosphorylated focal adhesion kinase and actin stress fiber formation 686:768 Interestingly, the levels of phosphorylated focal adhesion kinase and actin stress fiber formation were greatly enhanced in the S3-5 mutant. 28167607 10 75 theme cell 1472:1475 arg1 migration 1477:1485 cell migration 1472:1485 cell migration 1472:1485 These findings indicate that N-glycosylation on the β-propeller domain functions as a molecular switch to control the dynamics of α5β1 on the cell surface that in turn is required for optimum adhesion for cell migration. 28167607 1 76 theme cell 227:230 arg1 behavior 232:239 cell behavior 227:239 cell behavior 227:239 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 4 77 theme α5 546:547 arg1 cells 565:569 α5 knockout cancer cells 546:569 α5 knockout cancer cells 546:569 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 0 78 from α5 117:118 arg1 Characterization 53:68 Characterization 53:68 Characterization 53:68 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 0 78 from α5 117:118 arg1 Identification 34:47 Identification 34:47 Identification 34:47 A Key Regulator of Cell Adhesion: Identification and Characterization of Important N-Glycosylation Sites on Integrin α5 for Cell Migration. 28167607 5 79 with comparison 639:648 arg1 that 655:658 that 655:658 that 655:658 We found that the migration ability of S3-5 cells was decreased in comparison with that of the WT. 28167607 4 80 dep Wild-type 398:406 arg1 WT 409:410 WT 409:410 WT 409:410 Wild-type (WT) integrin α5 and N-glycosylation mutant S3-5 (sites 3 to 5) integrin α5, which contains fewer N-glycans, were stably reconstituted in α5 knockout cancer cells. 28167607 1 81 theme behavior 232:239 arg1 aspects 216:222 many fundamental aspects 199:222 many fundamental aspects 199:222 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 81 theme behavior 232:239 arg1 migration 270:278 migration 270:278 migration 270:278 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28167607 1 81 theme behavior 232:239 arg1 adhesion 257:264 cell adhesion 252:264 cell adhesion 252:264 The N-glycosylation of integrin α5β1 is thought to control many fundamental aspects of cell behavior, including cell adhesion and migration. 28486782 7 0 dep stability 828:836 arg1 the 824:826 the 824:826 the 824:826 Glycosylation has been shown to be important for modulating the stability and function of EGFR. 28486782 11 1 theme dimeric 1426:1432 arg1 interface 1434:1442 the dimeric interface 1422:1442 the dimeric interface 1422:1442 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 11 2 theme extracellular 1538:1550 arg1 domain 1552:1557 the EGFR extracellular domain 1529:1557 the EGFR extracellular domain 1529:1557 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 4 3 theme autoinhibited 363:375 arg1 conformation 386:397 the autoinhibited tethered conformation 359:397 the autoinhibited tethered conformation 359:397 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 9 4 theme extracellular 1203:1215 arg1 domain 1217:1222 EGFR extracellular domain 1198:1222 EGFR extracellular domain 1198:1222 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 1 5 theme several 150:156 arg1 cancers 158:164 several cancers 150:164 several cancers 150:164 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 12 6 theme glycosylation 1617:1629 arg1 possibility 1594:1604 the possibility 1590:1604 the possibility of altered glycosylation with functional consequences 1590:1658 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 8 7 theme factors 996:1002 arg1 binding 978:984 the binding 974:984 the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain 974:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 9 8 theme EGF 1233:1235 arg1 site 1245:1248 the EGF binding site 1229:1248 the EGF binding site 1229:1248 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 12 9 theme functional 1636:1645 arg1 consequences 1647:1658 functional consequences 1636:1658 functional consequences 1636:1658 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 3 10 theme Growth 237:242 arg1 factor 244:249 Growth factor 237:249 Growth factor (mostly epidermal growth factor or EGF) binding 237:297 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 2 11 theme EGFR 195:198 arg1 EGFR 195:198 EGFR 195:198 EGFR 195:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 2 11 theme EGFR 195:198 arg1 domain 185:190 The extracellular domain 167:190 The extracellular domain of EGFR 167:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 10 12 dep interactions 1321:1332 arg1 electrostatic 1335:1347 electrostatic 1335:1347 electrostatic 1335:1347 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 28486782 3 13 theme epidermal 259:267 arg1 factor 276:281 epidermal growth factor 259:281 epidermal growth factor 259:281 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 11 14 theme separated 1507:1515 arg1 epitopes 1517:1524 spatially separated epitopes 1497:1524 spatially separated epitopes of the EGFR extracellular domain 1497:1557 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 4 15 from conformation 386:397 arg1 transition 343:352 the transition 339:352 the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR 339:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 1 16 theme tyrosine 108:115 arg1 protein 124:130 a tyrosine kinase protein 106:130 a tyrosine kinase protein 106:130 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 16 theme tyrosine 108:115 arg1 receptor 87:94 The epidermal growth factor receptor 59:94 The epidermal growth factor receptor (EGFR) 59:101 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 12 17 theme data 1576:1579 arg1 Analysis 1560:1567 Analysis 1560:1567 Analysis of SNP data 1560:1579 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 11 18 theme EGFR 1533:1536 arg1 domain 1552:1557 the EGFR extracellular domain 1529:1557 the EGFR extracellular domain 1529:1557 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 8 19 theme monoclonal 1005:1014 arg1 antibodies 1016:1025 monoclonal antibodies 1005:1025 monoclonal antibodies 1005:1025 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 0 20 from Role 0:3 arg1 binding 50:56 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 5 21 theme homologous 589:598 arg1 receptor 600:607 another homologous receptor 581:607 another homologous receptor 581:607 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 4 22 theme form 444:447 arg1 conformation 414:425 an extended conformation 402:425 an extended conformation of the monomeric form of EGFR 402:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 11 23 theme distinct 1454:1461 arg1 roles 1463:1467 distinct roles 1454:1467 distinct roles 1454:1467 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 5 24 theme dimeric 543:549 arg1 conformation 551:562 a back-to-back dimeric conformation 528:562 a back-to-back dimeric conformation 528:562 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 1 25 theme growth 73:78 arg1 receptor 87:94 The epidermal growth factor receptor 59:94 The epidermal growth factor receptor (EGFR) 59:101 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 25 theme growth 73:78 arg1 protein 124:130 a tyrosine kinase protein 106:130 a tyrosine kinase protein 106:130 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 25 theme growth 73:78 arg1 EGFR 97:100 EGFR 97:100 EGFR 97:100 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 8 26 theme atomistic 866:874 arg1 simulations 879:889 atomistic MD simulations 866:889 atomistic MD simulations 866:889 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 4 27 theme EGFR 452:455 arg1 form 444:447 the monomeric form 430:447 the monomeric form of EGFR 430:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 10 28 theme factor 1278:1283 arg1 site 1293:1296 the growth factor binding site 1267:1296 the growth factor binding site 1267:1296 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 28486782 8 29 theme partners 1044:1051 arg1 binding 978:984 the binding 974:984 the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain 974:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 9 30 theme EGFR 1198:1201 arg1 domain 1217:1222 EGFR extracellular domain 1198:1222 EGFR extracellular domain 1198:1222 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 10 31 theme stronger 1312:1319 arg1 interactions 1321:1332 stronger interactions 1312:1332 stronger interactions (electrostatic) between the growth factor and EGFR 1312:1383 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 28486782 0 32 theme N-glycosylation 8:22 arg1 Role 0:3 Role 0:3 Role of N-glycosylation in EGFR ectodomain ligand binding. 0:57 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 0 33 theme EGFR 27:30 arg1 binding 50:56 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 8 34 theme EGFR 1070:1073 arg1 domain 1089:1094 the monomeric EGFR extracellular domain 1056:1094 the monomeric EGFR extracellular domain 1056:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 4 35 theme preformed 489:497 arg1 dimer 499:503 the flexible preformed dimer 476:503 the flexible preformed dimer 476:503 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 6 36 theme dimeric 744:750 arg1 interfaces 752:761 the dimeric interfaces 740:761 the dimeric interfaces 740:761 Several antibodies inhibit EGFR by targeting the growth factor binding site or the dimeric interfaces. 28486782 0 37 theme ligand 43:48 arg1 binding 50:56 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 8 38 theme EGFR 924:927 arg1 domain 943:948 the EGFR extracellular domain 920:948 the EGFR extracellular domain 920:948 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 6 39 theme factor 717:722 arg1 site 732:735 the growth factor binding site 706:735 the growth factor binding site 706:735 Several antibodies inhibit EGFR by targeting the growth factor binding site or the dimeric interfaces. 28486782 8 40 theme domain 943:948 arg1 N-glycosylation 901:915 N-glycosylation 901:915 N-glycosylation of the EGFR extracellular domain 901:948 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 8 41 gly N-glycosylation 901:915 arg1 domain 943:948 the EGFR extracellular domain 920:948 the EGFR extracellular domain 920:948 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 9 42 theme noncovalent 1149:1159 arg1 interactions 1161:1172 several noncovalent interactions 1141:1172 several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site 1141:1248 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 8 43 theme critical 956:963 arg1 roles 965:969 critical roles 956:969 critical roles 956:969 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 6 44 theme growth 710:715 arg1 site 732:735 the growth factor binding site 706:735 the growth factor binding site 706:735 Several antibodies inhibit EGFR by targeting the growth factor binding site or the dimeric interfaces. 28486782 5 45 theme dimeric 644:650 arg1 partner 652:658 the dimeric partner 640:658 the dimeric partner 640:658 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 12 46 with glycosylation 1617:1629 arg1 consequences 1647:1658 functional consequences 1636:1658 functional consequences 1636:1658 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 4 47 theme tethered 377:384 arg1 conformation 386:397 the autoinhibited tethered conformation 359:397 the autoinhibited tethered conformation 359:397 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 11 48 theme domain 1552:1557 arg1 epitopes 1517:1524 spatially separated epitopes 1497:1524 spatially separated epitopes of the EGFR extracellular domain 1497:1557 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 7 49 theme EGFR 854:857 arg1 function 842:849 function 842:849 function 842:849 Glycosylation has been shown to be important for modulating the stability and function of EGFR. 28486782 7 49 theme EGFR 854:857 arg1 stability 828:836 stability 828:836 stability 828:836 Glycosylation has been shown to be important for modulating the stability and function of EGFR. 28486782 8 50 theme growth 989:994 arg1 factors 996:1002 growth factors 989:1002 growth factors 989:1002 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 2 51 gly glycosylated 223:234 arg1 EGFR 195:198 EGFR 195:198 EGFR 195:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 2 51 gly glycosylated 223:234 arg1 domain 185:190 The extracellular domain 167:190 The extracellular domain of EGFR 167:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 14 52 dep © 1690:1690 arg1 Inc. 1716:1719 Inc. 1716:1719 Inc. 1716:1719 © 2017 Wiley Periodicals, Inc. 28486782 5 53 theme extracellular 616:628 arg1 domain 630:635 its extracellular domain 612:635 its extracellular domain as the dimeric partner 612:658 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 9 54 theme binding 1237:1243 arg1 site 1245:1248 the EGF binding site 1229:1248 the EGF binding site 1229:1248 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 3 55 theme factor 244:249 arg1 binding 291:297 Growth factor (mostly epidermal growth factor or EGF) binding 237:297 Growth factor (mostly epidermal growth factor or EGF) binding 237:297 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 4 56 theme monomeric 434:442 arg1 form 444:447 the monomeric form 430:447 the monomeric form of EGFR 430:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 12 57 theme altered 1609:1615 arg1 glycosylation 1617:1629 altered glycosylation 1609:1629 altered glycosylation with functional consequences 1609:1658 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 5 58 theme back-to-back 530:541 arg1 conformation 551:562 a back-to-back dimeric conformation 528:562 a back-to-back dimeric conformation 528:562 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 12 59 theme SNP 1572:1574 arg1 data 1576:1579 SNP data 1572:1579 SNP data 1572:1579 Analysis of SNP data suggests the possibility of altered glycosylation with functional consequences. 28486782 3 60 theme growth 269:274 arg1 factor 276:281 epidermal growth factor 259:281 epidermal growth factor 259:281 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 4 61 theme extended 405:412 arg1 conformation 414:425 an extended conformation 402:425 an extended conformation of the monomeric form of EGFR 402:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 11 62 theme antibodies 1483:1492 arg1 binding 1472:1478 binding 1472:1478 binding of antibodies to spatially separated epitopes of the EGFR extracellular domain 1472:1557 N-glycosylation also helps maintain the dimeric interface and plays distinct roles in binding of antibodies to spatially separated epitopes of the EGFR extracellular domain. 28486782 1 63 theme kinase 117:122 arg1 protein 124:130 a tyrosine kinase protein 106:130 a tyrosine kinase protein 106:130 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 63 theme kinase 117:122 arg1 receptor 87:94 The epidermal growth factor receptor 59:94 The epidermal growth factor receptor (EGFR) 59:101 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 9 64 dep glycans 1186:1192 arg1 the 1182:1184 the 1182:1184 the 1182:1184 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 10 65 theme growth 1271:1276 arg1 factor 1278:1283 growth factor 1271:1283 the growth factor binding site 1267:1296 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 28486782 8 66 theme antibodies 1016:1025 arg1 binding 978:984 the binding 974:984 the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain 974:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 5 67 theme Activated 506:514 arg1 EGFR 516:519 Activated EGFR 506:519 Activated EGFR 506:519 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 4 68 theme flexible 480:487 arg1 dimer 499:503 the flexible preformed dimer 476:503 the flexible preformed dimer 476:503 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 8 69 theme dimeric 1036:1042 arg1 partners 1044:1051 the dimeric partners 1032:1051 the dimeric partners 1032:1051 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 5 70 theme receptor 600:607 arg1 binding 570:576 binding 570:576 binding of another homologous receptor to its extracellular domain as the dimeric partner 570:658 Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner. 28486782 1 71 theme epidermal 63:71 arg1 receptor 87:94 The epidermal growth factor receptor 59:94 The epidermal growth factor receptor (EGFR) 59:101 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 71 theme epidermal 63:71 arg1 protein 124:130 a tyrosine kinase protein 106:130 a tyrosine kinase protein 106:130 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 71 theme epidermal 63:71 arg1 EGFR 97:100 EGFR 97:100 EGFR 97:100 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 10 72 theme binding 1285:1291 arg1 site 1293:1296 the growth factor binding site 1267:1296 the growth factor binding site 1267:1296 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 28486782 1 73 theme factor 80:85 arg1 receptor 87:94 The epidermal growth factor receptor 59:94 The epidermal growth factor receptor (EGFR) 59:101 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 73 theme factor 80:85 arg1 protein 124:130 a tyrosine kinase protein 106:130 a tyrosine kinase protein 106:130 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 1 73 theme factor 80:85 arg1 EGFR 97:100 EGFR 97:100 EGFR 97:100 The epidermal growth factor receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. 28486782 4 74 dep occurs 320:325 arg1 stabilizing 464:474 stabilizing 464:474 stabilizing the flexible preformed dimer 464:503 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 4 74 dep occurs 320:325 arg1 inducing 330:337 inducing 330:337 inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR 330:455 This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible preformed dimer. 28486782 8 75 theme monomeric 1060:1068 arg1 domain 1089:1094 the monomeric EGFR extracellular domain 1056:1094 the monomeric EGFR extracellular domain 1056:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 8 76 theme MD 876:877 arg1 simulations 879:889 atomistic MD simulations 866:889 atomistic MD simulations 866:889 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 6 77 theme binding 724:730 arg1 site 732:735 the growth factor binding site 706:735 the growth factor binding site 706:735 Several antibodies inhibit EGFR by targeting the growth factor binding site or the dimeric interfaces. 28486782 6 78 theme Several 661:667 arg1 antibodies 669:678 Several antibodies 661:678 Several antibodies 661:678 Several antibodies inhibit EGFR by targeting the growth factor binding site or the dimeric interfaces. 28486782 0 79 theme ectodomain 32:41 arg1 binding 50:56 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 8 80 theme extracellular 1075:1087 arg1 domain 1089:1094 the monomeric EGFR extracellular domain 1056:1094 the monomeric EGFR extracellular domain 1056:1094 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 0 81 gly N-glycosylation 8:22 arg1 ectodomain 32:41 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 0 81 gly N-glycosylation 8:22 arg1 ligand 43:48 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 0 81 gly N-glycosylation 8:22 arg1 EGFR 27:30 EGFR ectodomain ligand binding 27:56 EGFR ectodomain ligand binding 27:56 Role of N-glycosylation in EGFR ectodomain ligand binding. 28486782 9 82 theme interactions 1161:1172 arg1 formation 1128:1136 the formation 1124:1136 the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site 1124:1248 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 2 83 theme extracellular 171:183 arg1 EGFR 195:198 EGFR 195:198 EGFR 195:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 2 83 theme extracellular 171:183 arg1 domain 185:190 The extracellular domain 167:190 The extracellular domain of EGFR 167:198 The extracellular domain of EGFR is known to be heavily glycosylated. 28486782 9 84 theme several 1141:1147 arg1 interactions 1161:1172 several noncovalent interactions 1141:1172 several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site 1141:1248 N-glycosylation results in the formation of several noncovalent interactions between the glycans and EGFR extracellular domain near the EGF binding site. 28486782 8 85 theme extracellular 929:941 arg1 domain 943:948 the EGFR extracellular domain 920:948 the EGFR extracellular domain 920:948 Here, atomistic MD simulations show that N-glycosylation of the EGFR extracellular domain plays critical roles in the binding of growth factors, monoclonal antibodies, and the dimeric partners to the monomeric EGFR extracellular domain. 28486782 3 86 dep factor 244:249 arg1 EGF 286:288 EGF 286:288 EGF 286:288 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 3 86 dep factor 244:249 arg1 factor 276:281 epidermal growth factor 259:281 epidermal growth factor 259:281 Growth factor (mostly epidermal growth factor or EGF) binding activates EGFR. 28486782 10 87 theme growth 1362:1367 arg1 factor 1369:1374 the growth factor 1358:1374 the growth factor 1358:1374 This stabilizes the growth factor binding site, resulting in stronger interactions (electrostatic) between the growth factor and EGFR. 26488311 7 0 theme low 1169:1171 arg1 atrophy 1173:1179 low atrophy 1169:1179 low atrophy 1169:1179 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 7 1 theme β64N 1072:1075 arg1 glycoforms 1077:1086 Eight β64N glycoforms 1066:1086 Eight β64N glycoforms 1066:1086 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 5 2 theme liquid 682:687 arg1 LC-MS/MS 730:737 LC-MS/MS 730:737 LC-MS/MS 730:737 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 2 theme liquid 682:687 arg1 spectrometry 716:727 liquid chromatography-tandem mass spectrometry 682:727 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 682:738 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 3 3 theme known 355:359 arg1 sites 377:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 8 4 gly glycoforms 1402:1411 arg1 clusterin 1392:1400 specific clusterin glycoforms 1383:1411 specific clusterin glycoforms 1383:1411 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 4 5 theme clusterin 547:555 arg1 glycopeptides 557:569 clusterin glycopeptides 547:569 clusterin glycopeptides 547:569 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 5 6 theme mass 711:714 arg1 LC-MS/MS 730:737 LC-MS/MS 730:737 LC-MS/MS 730:737 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 6 theme mass 711:714 arg1 spectrometry 716:727 liquid chromatography-tandem mass spectrometry 682:727 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 682:738 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 7 theme low 896:898 arg1 n 921:921 n = 13 921:926 n = 13 921:926 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 7 theme low 896:898 arg1 atrophy 912:918 low hippocampal atrophy 896:918 low hippocampal atrophy (n = 13) 896:927 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 8 theme hippocampal 900:910 arg1 n 921:921 n = 13 921:926 n = 13 921:926 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 8 theme hippocampal 900:910 arg1 atrophy 912:918 low hippocampal atrophy 896:918 low hippocampal atrophy (n = 13) 896:927 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 8 9 theme novel 1322:1326 arg1 workflow 1339:1346 a novel and robust workflow 1320:1346 a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers 1320:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 5 10 theme hippocampal 938:948 arg1 atrophy 950:956 high hippocampal atrophy 933:956 high hippocampal atrophy (n = 14) 933:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 10 theme hippocampal 938:948 arg1 n 959:959 n = 14 959:964 n = 14 959:964 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 1 11 dep found 147:151 arg1 associated 153:162 associated 153:162 associated with hippocampal atrophy 153:187 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 7 12 with patients 1117:1124 arg1 atrophy 1136:1142 high atrophy 1131:1142 high atrophy 1131:1142 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 5 13 theme atrophy 950:956 arg1 subjects 884:891 subjects 884:891 subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 884:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 2 14 from plasma 232:237 arg1 glycosylation 194:206 The glycosylation 190:206 The glycosylation of clusterin from human plasma 190:237 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 3 15 theme beta 463:466 arg1 subunit 468:474 the beta subunit 459:474 the beta subunit (β64N, β127N, and β147N) 459:499 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 16 theme N-glycosylation 361:375 arg1 sites 377:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 17 from three 396:400 arg1 subunit 415:421 the alpha subunit 405:421 the alpha subunit (α64N, α81N and α123N) 405:444 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 5 18 gly glycosylation 830:842 arg2 sites 844:848 the different glycosylation sites 816:848 the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 816:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 6 19 theme β64N 992:995 arg1 site 997:1000 the β64N site 988:1000 the β64N site 988:1000 In our pilot study, the β64N site shows the most significant regulations between clinical groups. 26488311 2 20 theme spectrometry 297:308 arg1 analysis 337:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 7 21 theme disease 1276:1282 arg1 markers 1289:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 1 22 theme hippocampal 169:179 arg1 atrophy 181:187 hippocampal atrophy 169:187 hippocampal atrophy 169:187 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 4 23 theme glycopeptides 557:569 arg1 characterization 527:542 More detailed structural characterization 502:542 More detailed structural characterization of clusterin glycopeptides 502:569 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 4 24 dep detailed 507:514 arg1 structural 516:525 structural 516:525 structural 516:525 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 3 25 dep subunit 415:421 arg1 α123N 439:443 α123N 439:443 α123N 439:443 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 25 dep subunit 415:421 arg1 α81N 430:433 α81N 430:433 α81N 430:433 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 25 dep subunit 415:421 arg1 α64N 424:427 α64N 424:427 α64N 424:427 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 2 26 theme mass 292:295 arg1 MS 311:312 MS 311:312 MS 311:312 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 2 26 theme mass 292:295 arg1 spectrometry 297:308 mass spectrometry 292:308 mass spectrometry (MS)-based glycoproteomics analysis 292:344 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 4 27 theme peptides 634:641 arg1 presence 609:616 the presence 605:616 the presence of glycosylated peptides and their corresponding glycans 605:673 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 7 28 theme high 1131:1134 arg1 atrophy 1136:1142 high atrophy 1131:1142 high atrophy 1131:1142 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 6 29 theme significant 1017:1027 arg1 regulations 1029:1039 the most significant regulations 1008:1039 the most significant regulations between clinical groups 1008:1063 In our pilot study, the β64N site shows the most significant regulations between clinical groups. 26488311 3 30 dep subunit 468:474 arg1 β127N 483:487 β127N 483:487 β127N 483:487 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 30 dep subunit 468:474 arg1 β147N 494:498 β147N 494:498 β147N 494:498 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 30 dep subunit 468:474 arg1 β64N 477:480 β64N 477:480 β64N 477:480 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 7 31 theme isoforms 1226:1233 arg1 utility 1205:1211 the utility 1201:1211 the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers 1201:1295 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 4 32 theme corresponding 653:665 arg1 glycans 667:673 their corresponding glycans 647:673 their corresponding glycans 647:673 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 4 33 theme detailed 507:514 arg1 characterization 527:542 More detailed structural characterization 502:542 More detailed structural characterization of clusterin glycopeptides 502:569 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 4 34 gly glycosylated 621:632 arg1 peptides 634:641 glycosylated peptides 621:641 glycosylated peptides 621:641 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 4 35 gly glycopeptides 557:569 arg2 glycopeptides 557:569 clusterin glycopeptides 547:569 clusterin glycopeptides 547:569 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 2 36 theme glycoproteomics 321:335 arg1 analysis 337:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 0 37 theme Plasma 23:28 arg1 Clusterin 30:38 Human Plasma Clusterin 17:38 Human Plasma Clusterin 17:38 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 5 38 theme chromatography-tandem 689:709 arg1 LC-MS/MS 730:737 LC-MS/MS 730:737 LC-MS/MS 730:737 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 38 theme chromatography-tandem 689:709 arg1 spectrometry 716:727 liquid chromatography-tandem mass spectrometry 682:727 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 682:738 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 39 theme = 923:923 arg1 n 921:921 n = 13 921:926 n = 13 921:926 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 39 theme = 923:923 arg1 atrophy 912:918 low hippocampal atrophy 896:918 low hippocampal atrophy (n = 13) 896:927 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 2 40 theme -based 314:319 arg1 analysis 337:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 mass spectrometry (MS)-based glycoproteomics analysis 292:344 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 5 41 theme different 820:828 arg1 sites 844:848 the different glycosylation sites 816:848 the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 816:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 0 42 theme Human 17:21 arg1 Clusterin 30:38 Human Plasma Clusterin 17:38 Human Plasma Clusterin 17:38 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 1 43 gly glycosylated 108:119 arg1 peptides 121:128 Specific glycosylated peptides 99:128 Specific glycosylated peptides of clusterin 99:141 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 8 44 theme glycoforms 1402:1411 arg1 verification 1367:1378 rapid verification 1361:1378 rapid verification of specific clusterin glycoforms with utility as AD biomarkers 1361:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 1 45 theme Specific 99:106 arg1 peptides 121:128 Specific glycosylated peptides 99:128 Specific glycosylated peptides of clusterin 99:141 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 5 46 theme glycosylation 830:842 arg1 sites 844:848 the different glycosylation sites 816:848 the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 816:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 8 47 theme clusterin 1392:1400 arg1 glycoforms 1402:1411 specific clusterin glycoforms 1383:1411 specific clusterin glycoforms 1383:1411 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 0 48 gly Glycosylation 0:12 arg1 Clusterin 30:38 Human Plasma Clusterin 17:38 Human Plasma Clusterin 17:38 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 4 49 theme glycosylated 621:632 arg1 peptides 634:641 glycosylated peptides 621:641 glycosylated peptides 621:641 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 0 50 theme Clusterin 30:38 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of Human Plasma Clusterin 0:38 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 3 51 gly N-glycosylation 361:375 arg2 sites 377:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites 347:381 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 51 gly N-glycosylation 361:375 arg2 six 351:353 six 351:353 six 351:353 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 8 52 theme robust 1332:1337 arg1 workflow 1339:1346 a novel and robust workflow 1320:1346 a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers 1320:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 5 53 from sites 844:848 arg1 clusterin 860:868 plasma clusterin 853:868 plasma clusterin 853:868 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 7 54 theme prognostic 1253:1262 arg1 markers 1289:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 3 55 theme alpha 409:413 arg1 subunit 415:421 the alpha subunit 405:421 the alpha subunit (α64N, α81N and α123N) 405:444 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 0 56 theme Candidate 55:63 arg1 Biomarker 65:73 a Novel Candidate Biomarker 47:73 a Novel Candidate Biomarker of Alzheimer's Disease 47:96 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 1 57 theme glycosylated 108:119 arg1 peptides 121:128 Specific glycosylated peptides 99:128 Specific glycosylated peptides of clusterin 99:141 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 8 58 with verification 1367:1378 arg1 utility 1418:1424 utility 1418:1424 utility as AD biomarkers 1418:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 8 59 theme AD 1429:1430 arg1 biomarkers 1432:1441 AD biomarkers 1429:1441 AD biomarkers 1429:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 2 60 theme human 226:230 arg1 plasma 232:237 human plasma 226:237 human plasma 226:237 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 3 61 from three 450:454 arg1 subunit 468:474 the beta subunit 459:474 the beta subunit (β64N, β127N, and β147N) 459:499 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 7 62 theme diagnostic 1238:1247 arg1 markers 1289:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 diagnostic and prognostic Alzheimer's disease (AD) markers 1238:1295 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 2 63 theme clusterin 211:219 arg1 glycosylation 194:206 The glycosylation 190:206 The glycosylation of clusterin from human plasma 190:237 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 5 64 theme = 961:961 arg1 atrophy 950:956 high hippocampal atrophy 933:956 high hippocampal atrophy (n = 14) 933:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 64 theme = 961:961 arg1 n 959:959 n = 14 959:964 n = 14 959:964 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 7 65 theme clusterin 1216:1224 arg1 isoforms 1226:1233 clusterin isoforms 1216:1233 clusterin isoforms 1216:1233 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 3 66 dep covered 387:393 arg1 three 450:454 three 450:454 three 450:454 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 3 66 dep covered 387:393 arg1 three 396:400 three 396:400 three 396:400 All six known N-glycosylation sites are covered, three in the alpha subunit (α64N, α81N and α123N) and three in the beta subunit (β64N, β127N, and β147N). 26488311 5 67 theme atrophy 912:918 arg1 subjects 884:891 subjects 884:891 subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 884:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 8 68 theme rapid 1361:1365 arg1 verification 1367:1378 rapid verification 1361:1378 rapid verification of specific clusterin glycoforms with utility as AD biomarkers 1361:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 5 69 theme plasma 853:858 arg1 clusterin 860:868 plasma clusterin 853:868 plasma clusterin 853:868 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 1 70 theme clusterin 133:141 arg1 peptides 121:128 Specific glycosylated peptides 99:128 Specific glycosylated peptides of clusterin 99:141 Specific glycosylated peptides of clusterin are found associated with hippocampal atrophy. 26488311 2 71 gly glycosylation 194:206 arg1 clusterin 211:219 clusterin 211:219 clusterin 211:219 The glycosylation of clusterin from human plasma was comprehensively analyzed and characterized using mass spectrometry (MS)-based glycoproteomics analysis. 26488311 6 72 theme pilot 975:979 arg1 study 981:985 our pilot study 971:985 our pilot study 971:985 In our pilot study, the β64N site shows the most significant regulations between clinical groups. 26488311 6 73 theme clinical 1049:1056 arg1 groups 1058:1063 clinical groups 1049:1063 clinical groups 1049:1063 In our pilot study, the β64N site shows the most significant regulations between clinical groups. 26488311 8 74 theme specific 1383:1390 arg1 glycoforms 1402:1411 specific clusterin glycoforms 1383:1411 specific clusterin glycoforms 1383:1411 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 26488311 7 75 gly glycoforms 1077:1086 arg1 β64N 1072:1075 Eight β64N glycoforms 1066:1086 Eight β64N glycoforms 1066:1086 Eight β64N glycoforms are significantly reduced in patients with high atrophy compared with those with low atrophy, which demonstrates the utility of clusterin isoforms as diagnostic and prognostic Alzheimer's disease (AD) markers. 26488311 5 76 theme high 933:936 arg1 atrophy 950:956 high hippocampal atrophy 933:956 high hippocampal atrophy (n = 14) 933:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 76 theme high 933:936 arg1 n 959:959 n = 14 959:964 n = 14 959:964 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 5 77 from differences 764:774 arg1 glycoforms 783:792 the glycoforms 779:792 the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14) 779:965 Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we have determined the differences in the glycoforms associated at each of the different glycosylation sites in plasma clusterin obtained from subjects of low hippocampal atrophy (n = 13) and high hippocampal atrophy (n = 14). 26488311 0 78 theme Novel 49:53 arg1 Biomarker 65:73 a Novel Candidate Biomarker 47:73 a Novel Candidate Biomarker of Alzheimer's Disease 47:96 Glycosylation of Human Plasma Clusterin Yields a Novel Candidate Biomarker of Alzheimer's Disease. 26488311 4 79 theme glycans 667:673 arg1 presence 609:616 the presence 605:616 the presence of glycosylated peptides and their corresponding glycans 605:673 More detailed structural characterization of clusterin glycopeptides was also performed, demonstrating the presence of glycosylated peptides and their corresponding glycans. 26488311 8 80 theme suitable 1348:1355 arg1 workflow 1339:1346 a novel and robust workflow 1320:1346 a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers 1320:1441 These results provide a novel and robust workflow suitable for rapid verification of specific clusterin glycoforms with utility as AD biomarkers. 28446609 3 0 theme asparagine 622:631 arg1 residues 633:640 the glycosylated asparagine residues 605:640 the glycosylated asparagine residues 605:640 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 1 1 theme ∼25 217:219 arg1 sites 235:239 ∼25 glycosylation sites 217:239 ∼25 glycosylation sites 217:239 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 2 from located 260:266 arg1 V1/V2 275:279 the V1/V2 and V3 variable loops 271:301 V1/V2 275:279 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 2 from located 260:266 arg1 loops 297:301 the V1/V2 and V3 variable loops 271:301 loops 297:301 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 2 from located 260:266 arg1 others 311:316 the others 307:316 the others in the remaining core gp120 region 307:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 8 3 theme glycosylated 1706:1717 arg1 Env 1719:1721 fully glycosylated Env 1700:1721 fully glycosylated Env 1700:1721 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 1 4 theme sites 235:239 arg1 located 260:266 located 260:266 located 260:266 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 3 5 gly glycosylation 550:562 arg2 sites 564:568 glycosylation sites 550:568 glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines 550:687 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 3 6 theme similar 658:664 arg1 glutamines 666:675 glutamines 666:675 glutamines 666:675 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 6 7 theme primary 1441:1447 arg1 role 1449:1452 their likely primary role 1428:1452 their likely primary role 1428:1452 Our observations demonstrate that core gp120 glycans are not essential for folding, and hence their likely primary role is enabling immune evasion. 28446609 1 8 from loops 297:301 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 8 from loops 297:301 arg1 located 260:266 located 260:266 located 260:266 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 9 theme HIV-1 161:165 arg1 protein 182:188 the HIV-1 envelope (Env) protein 157:188 the HIV-1 envelope (Env) protein 157:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 0 10 theme trimer 97:102 arg1 formation 104:112 native trimer formation 90:112 native trimer formation 90:112 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 5 11 theme Env 1062:1064 arg1 variants 1066:1073 properly cleaved HIV-1 Env variants 1039:1073 properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01 1039:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 12 with information 961:971 arg1 design 995:1000 structure-guided design 978:1000 structure-guided design 978:1000 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 12 with information 961:971 arg1 display 1020:1026 yeast surface display 1006:1026 yeast surface display 1006:1026 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 13 theme several 1197:1203 arg1 antibodies 1272:1281 trimer-specific antibodies 1256:1281 trimer-specific antibodies 1256:1281 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 13 theme several 1197:1203 arg1 bNAbs 1238:1242 bNAbs 1238:1242 bNAbs 1238:1242 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 13 theme several 1197:1203 arg1 antibodies 1226:1235 several broadly neutralizing antibodies 1197:1235 several broadly neutralizing antibodies (bNAbs) 1197:1243 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 13 theme several 1197:1203 arg1 version 1307:1313 a germline-reverted version 1287:1313 a germline-reverted version of the bNAb VRC01 1287:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 1 14 theme remaining 325:333 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 6 15 theme likely 1434:1439 arg1 role 1449:1452 their likely primary role 1428:1452 their likely primary role 1428:1452 Our observations demonstrate that core gp120 glycans are not essential for folding, and hence their likely primary role is enabling immune evasion. 28446609 9 16 theme gp120 1778:1782 arg1 glycosylation 1784:1796 core gp120 glycosylation 1773:1796 core gp120 glycosylation 1773:1796 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 1 17 theme gp120 340:344 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 2 18 theme gp120 490:494 arg1 folding 479:485 proper folding 472:485 proper folding of gp120 472:494 Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. 28446609 5 19 theme broadly 1205:1211 arg1 antibodies 1272:1281 trimer-specific antibodies 1256:1281 trimer-specific antibodies 1256:1281 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 19 theme broadly 1205:1211 arg1 bNAbs 1238:1242 bNAbs 1238:1242 bNAbs 1238:1242 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 19 theme broadly 1205:1211 arg1 antibodies 1226:1235 several broadly neutralizing antibodies 1197:1235 several broadly neutralizing antibodies (bNAbs) 1197:1243 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 19 theme broadly 1205:1211 arg1 version 1307:1313 a germline-reverted version 1287:1313 a germline-reverted version of the bNAb VRC01 1287:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 20 theme cleaved 1048:1054 arg1 variants 1066:1073 properly cleaved HIV-1 Env variants 1039:1073 properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01 1039:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 3 21 theme glycosylation 550:562 arg1 sites 564:568 glycosylation sites 550:568 glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines 550:687 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 4 22 from sites 755:759 arg1 mutations 719:727 such mutations 714:727 such mutations at multiple glycosylation sites 714:759 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 0 23 theme subunit 48:54 arg1 gp120 64:68 the HIV-1 envelope subunit protein gp120 29:68 the HIV-1 envelope subunit protein gp120 29:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 0 24 theme gp120 64:68 arg1 core 21:24 the core 17:24 the core of the HIV-1 envelope subunit protein gp120 17:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 1 25 theme V3 285:286 arg1 loops 297:301 the V1/V2 and V3 variable loops 271:301 loops 297:301 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 26 theme gp120 140:144 arg1 subunit 146:152 The gp120 subunit 136:152 The gp120 subunit of the HIV-1 envelope (Env) protein 136:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 6 27 theme gp120 1373:1377 arg1 glycans 1379:1385 core gp120 glycans 1368:1385 core gp120 glycans 1368:1385 Our observations demonstrate that core gp120 glycans are not essential for folding, and hence their likely primary role is enabling immune evasion. 28446609 1 28 gly glycosylated 201:212 arg2 sites 235:239 ∼25 glycosylation sites 217:239 ∼25 glycosylation sites 217:239 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 28 gly glycosylated 201:212 arg1 subunit 146:152 The gp120 subunit 136:152 The gp120 subunit of the HIV-1 envelope (Env) protein 136:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 29 from V1/V2 275:279 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 29 from V1/V2 275:279 arg1 located 260:266 located 260:266 located 260:266 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 5 30 theme neutralizing 1213:1224 arg1 antibodies 1272:1281 trimer-specific antibodies 1256:1281 trimer-specific antibodies 1256:1281 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 30 theme neutralizing 1213:1224 arg1 bNAbs 1238:1242 bNAbs 1238:1242 bNAbs 1238:1242 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 30 theme neutralizing 1213:1224 arg1 antibodies 1226:1235 several broadly neutralizing antibodies 1197:1235 several broadly neutralizing antibodies (bNAbs) 1197:1243 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 30 theme neutralizing 1213:1224 arg1 version 1307:1313 a germline-reverted version 1287:1313 a germline-reverted version of the bNAb VRC01 1287:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 31 theme core 1092:1095 arg1 glycans 1103:1109 all 15 core gp120 glycans 1085:1109 all 15 core gp120 glycans 1085:1109 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 1 32 from others 311:316 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 32 from others 311:316 arg1 located 260:266 located 260:266 located 260:266 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 5 33 theme viral 1167:1171 arg1 infectivity 1173:1183 multiple-cycle viral infectivity 1152:1183 multiple-cycle viral infectivity 1152:1183 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 34 theme alternative 914:924 arg1 approach 926:933 an alternative approach 911:933 an alternative approach 911:933 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 0 35 gly Glycosylation 0:12 arg1 core 21:24 the core 17:24 the core of the HIV-1 envelope subunit protein gp120 17:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 2 36 theme host 407:410 arg1 system 419:424 the host immune system 403:424 the host immune system 403:424 Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. 28446609 8 37 theme Glycan-deficient 1554:1569 arg1 immunogens 1610:1619 priming immunogens 1602:1619 priming immunogens 1602:1619 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 37 theme Glycan-deficient 1554:1569 arg1 derivatives 1575:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 5 38 theme VRC01 1327:1331 arg1 antibodies 1272:1281 trimer-specific antibodies 1256:1281 trimer-specific antibodies 1256:1281 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 38 theme VRC01 1327:1331 arg1 version 1307:1313 a germline-reverted version 1287:1313 a germline-reverted version of the bNAb VRC01 1287:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 39 theme conformational 1123:1136 arg1 integrity 1138:1146 conformational integrity 1123:1146 conformational integrity 1123:1146 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 40 theme surface 1012:1018 arg1 display 1020:1026 yeast surface display 1006:1026 yeast surface display 1006:1026 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 0 41 theme HIV-1 33:37 arg1 gp120 64:68 the HIV-1 envelope subunit protein gp120 29:68 the HIV-1 envelope subunit protein gp120 29:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 7 42 theme glycan 1504:1509 arg1 approach 1519:1526 our glycan removal approach 1500:1526 our glycan removal approach 1500:1526 We also show that our glycan removal approach is not strain restricted. 28446609 9 43 theme folded 1856:1861 arg1 derivatives 1871:1881 glycan-free folded protein derivatives 1844:1881 glycan-free folded protein derivatives 1844:1881 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 8 44 used used 1594:1597 arg2 derivatives 1575:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 44 used used 1594:1597 arg2 immunogens 1610:1619 priming immunogens 1602:1619 priming immunogens 1602:1619 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 4 45 theme multiple 732:739 arg1 sites 755:759 multiple glycosylation sites 732:759 multiple glycosylation sites 732:759 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 1 46 theme protein 182:188 arg1 subunit 146:152 The gp120 subunit 136:152 The gp120 subunit of the HIV-1 envelope (Env) protein 136:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 5 47 theme structure-guided 978:993 arg1 design 995:1000 structure-guided design 978:1000 structure-guided design 978:1000 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 0 48 theme envelope 39:46 arg1 gp120 64:68 the HIV-1 envelope subunit protein gp120 29:68 the HIV-1 envelope subunit protein gp120 29:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 6 49 theme immune 1466:1471 arg1 evasion 1473:1479 immune evasion 1466:1479 immune evasion 1466:1479 Our observations demonstrate that core gp120 glycans are not essential for folding, and hence their likely primary role is enabling immune evasion. 28446609 4 50 theme reduced 824:830 arg1 binding 832:838 significantly reduced binding 810:838 significantly reduced binding to both neutralizing and non-neutralizing antibodies 810:891 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 3 51 from sites 564:568 arg1 Env 573:575 Env 573:575 Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines 573:687 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 8 52 theme germlines 1685:1693 arg1 germlines 1685:1693 germlines 1685:1693 germlines 1685:1693 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 52 theme germlines 1685:1693 arg1 set 1678:1680 a more divergent set 1661:1680 a more divergent set of germlines 1661:1693 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 1 53 theme glycosylation 221:233 arg1 sites 235:239 ∼25 glycosylation sites 217:239 ∼25 glycosylation sites 217:239 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 2 54 theme viral 504:508 arg1 infectivity 510:520 viral infectivity 504:520 viral infectivity 504:520 Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. 28446609 2 55 theme immune 412:417 arg1 system 419:424 the host immune system 403:424 the host immune system 403:424 Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. 28446609 1 56 theme envelope 167:174 arg1 protein 182:188 the HIV-1 envelope (Env) protein 157:188 the HIV-1 envelope (Env) protein 157:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 8 57 gly glycosylated 1706:1717 arg1 Env 1719:1721 fully glycosylated Env 1700:1721 fully glycosylated Env 1700:1721 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 0 58 theme viral 117:121 arg1 infectivity 123:133 viral infectivity 117:133 viral infectivity 117:133 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 1 59 theme Env 177:179 arg1 protein 182:188 the HIV-1 envelope (Env) protein 157:188 the HIV-1 envelope (Env) protein 157:188 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 60 theme core 335:338 arg1 region 346:351 the remaining core gp120 region 321:351 the remaining core gp120 region 321:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 4 61 theme neutralizing 848:859 arg1 antibodies 882:891 both neutralizing and non-neutralizing antibodies 843:891 antibodies 882:891 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 3 62 theme glycosylated 609:620 arg1 residues 633:640 the glycosylated asparagine residues 605:640 the glycosylated asparagine residues 605:640 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 4 63 theme non-neutralizing 865:880 arg1 antibodies 882:891 both neutralizing and non-neutralizing antibodies 843:891 antibodies 882:891 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 5 64 theme germline-reverted 1289:1305 arg1 version 1307:1313 a germline-reverted version 1287:1313 a germline-reverted version of the bNAb VRC01 1287:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 0 65 theme protein 56:62 arg1 gp120 64:68 the HIV-1 envelope subunit protein gp120 29:68 the HIV-1 envelope subunit protein gp120 29:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 5 66 theme HIV-1 1056:1060 arg1 variants 1066:1073 properly cleaved HIV-1 Env variants 1039:1073 properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01 1039:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 3 67 gly glycosylated 609:620 arg1 residues 633:640 the glycosylated asparagine residues 605:640 the glycosylated asparagine residues 605:640 Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or alanines. 28446609 2 68 theme proper 472:477 arg1 folding 479:485 proper folding 472:485 proper folding of gp120 472:494 Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. 28446609 5 69 theme trimer-specific 1256:1270 arg1 antibodies 1272:1281 trimer-specific antibodies 1256:1281 trimer-specific antibodies 1256:1281 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 1 70 theme variable 288:295 arg1 loops 297:301 the V1/V2 and V3 variable loops 271:301 loops 297:301 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 9 71 theme core 1773:1776 arg1 glycosylation 1784:1796 core gp120 glycosylation 1773:1796 core gp120 glycosylation 1773:1796 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 0 72 theme native 90:95 arg1 formation 104:112 native trimer formation 90:112 native trimer formation 90:112 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 5 73 theme evolutionary 948:959 arg1 information 961:971 evolutionary information 948:971 evolutionary information with structure-guided design and yeast surface display 948:1026 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 9 74 theme glycosylation 1784:1796 arg1 role 1765:1768 the role 1761:1768 the role of core gp120 glycosylation 1761:1796 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 4 75 theme such 714:717 arg1 mutations 719:727 such mutations 714:727 such mutations at multiple glycosylation sites 714:759 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 6 76 theme core 1368:1371 arg1 glycans 1379:1385 core gp120 glycans 1368:1385 core gp120 glycans 1368:1385 Our observations demonstrate that core gp120 glycans are not essential for folding, and hence their likely primary role is enabling immune evasion. 28446609 5 77 theme gp120 1097:1101 arg1 glycans 1103:1109 all 15 core gp120 glycans 1085:1109 all 15 core gp120 glycans 1085:1109 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 1 78 located located 260:266 arg1 V1/V2 275:279 the V1/V2 and V3 variable loops 271:301 V1/V2 275:279 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 78 located located 260:266 arg2 ∼7-8 251:254 ∼7-8 251:254 ∼7-8 251:254 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 78 located located 260:266 arg1 loops 297:301 the V1/V2 and V3 variable loops 271:301 loops 297:301 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 1 78 located located 260:266 arg1 others 311:316 the others 307:316 the others in the remaining core gp120 region 307:351 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 0 79 theme core 21:24 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 0:68 Glycosylation of the core of the HIV-1 envelope subunit protein gp120 is not required for native trimer formation or viral infectivity. 28446609 1 80 gly glycosylation 221:233 arg2 sites 235:239 ∼25 glycosylation sites 217:239 ∼25 glycosylation sites 217:239 The gp120 subunit of the HIV-1 envelope (Env) protein is heavily glycosylated at ∼25 glycosylation sites, of which ∼7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. 28446609 9 81 theme general 1815:1821 arg1 method 1823:1828 a general method 1813:1828 a general method for designing glycan-free folded protein derivatives 1813:1881 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 8 82 theme Env 1571:1573 arg1 immunogens 1610:1619 priming immunogens 1602:1619 priming immunogens 1602:1619 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 82 theme Env 1571:1573 arg1 derivatives 1575:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 5 83 theme bNAb 1322:1325 arg1 VRC01 1327:1331 the bNAb VRC01 1318:1331 the bNAb VRC01 1318:1331 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 5 84 theme yeast 1006:1010 arg1 display 1020:1026 yeast surface display 1006:1026 yeast surface display 1006:1026 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 4 85 gly glycosylation 741:753 arg2 sites 755:759 multiple glycosylation sites 732:759 multiple glycosylation sites 732:759 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 9 86 theme glycan-free 1844:1854 arg1 derivatives 1871:1881 glycan-free folded protein derivatives 1844:1881 glycan-free folded protein derivatives 1844:1881 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 4 87 theme viral 778:782 arg1 infectivity 784:794 viral infectivity 778:794 viral infectivity 778:794 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 28446609 5 88 theme multiple-cycle 1152:1165 arg1 infectivity 1173:1183 multiple-cycle viral infectivity 1152:1183 multiple-cycle viral infectivity 1152:1183 Therefore, using an alternative approach, we combined evolutionary information with structure-guided design and yeast surface display to produce properly cleaved HIV-1 Env variants that lack all 15 core gp120 glycans, yet retain conformational integrity and multiple-cycle viral infectivity and bind to several broadly neutralizing antibodies (bNAbs), including trimer-specific antibodies and a germline-reverted version of the bNAb VRC01. 28446609 7 89 theme removal 1511:1517 arg1 approach 1519:1526 our glycan removal approach 1500:1526 our glycan removal approach 1500:1526 We also show that our glycan removal approach is not strain restricted. 28446609 9 90 theme protein 1863:1869 arg1 derivatives 1871:1881 glycan-free folded protein derivatives 1844:1881 glycan-free folded protein derivatives 1844:1881 In conclusion, these results clarify the role of core gp120 glycosylation and illustrate a general method for designing glycan-free folded protein derivatives. 28446609 8 91 theme priming 1602:1608 arg1 immunogens 1610:1619 priming immunogens 1602:1619 priming immunogens 1602:1619 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 91 theme priming 1602:1608 arg1 derivatives 1575:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives 1554:1585 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 92 theme divergent 1668:1676 arg1 germlines 1685:1693 germlines 1685:1693 germlines 1685:1693 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 8 92 theme divergent 1668:1676 arg1 set 1678:1680 a more divergent set 1661:1680 a more divergent set of germlines 1661:1693 Glycan-deficient Env derivatives can be used as priming immunogens because they should engage and activate a more divergent set of germlines than fully glycosylated Env. 28446609 4 93 theme glycosylation 741:753 arg1 sites 755:759 multiple glycosylation sites 732:759 multiple glycosylation sites 732:759 Here, we confirmed that such mutations at multiple glycosylation sites greatly diminish viral infectivity and result in significantly reduced binding to both neutralizing and non-neutralizing antibodies. 26482295 8 0 theme biocatalytic 1592:1603 arg1 diversity 1605:1613 previously unavailable biocatalytic diversity 1569:1613 previously unavailable biocatalytic diversity 1569:1613 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 2 1 theme diverse 384:390 arg1 structures 399:408 many diverse glycan structures 379:408 many diverse glycan structures 379:408 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 8 2 theme unavailable 1580:1590 arg1 diversity 1605:1613 previously unavailable biocatalytic diversity 1569:1613 previously unavailable biocatalytic diversity 1569:1613 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 1 3 theme specific 287:294 arg1 residues 307:314 specific asparagine residues 287:314 specific asparagine residues in target proteins 287:333 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 2 4 theme many 379:382 arg1 structures 399:408 many diverse glycan structures 379:408 many diverse glycan structures 379:408 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 1 5 theme asparagine 296:305 arg1 residues 307:314 specific asparagine residues 287:314 specific asparagine residues in target proteins 287:333 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 3 6 with preferences 610:620 arg1 variation 660:668 natural sequence variation 643:668 natural sequence variation compared to CjPglB 643:687 Here, we investigated the acceptor-site preferences for 23 homologs with natural sequence variation compared to CjPglB. 26482295 7 7 theme native 1391:1396 arg1 sequon 1406:1411 its native 'QYNST' sequon 1387:1411 its native 'QYNST' sequon 1387:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 0 8 gly N-glycosylation 80:94 arg2 site 96:99 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 0 8 gly N-glycosylation 80:94 arg2 preferences 101:111 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 8 9 theme own 1476:1478 arg1 rules 1480:1484 their own rules 1470:1484 their own rules for acceptor-site specificity 1470:1514 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 6 10 theme acceptor-site 1231:1243 arg1 preferences 1245:1255 acceptor-site preferences 1231:1255 acceptor-site preferences 1231:1255 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 7 11 theme bacterial 1315:1323 arg1 OST 1325:1327 the only bacterial OST 1306:1327 the only bacterial OST 1306:1327 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 12 theme only 1310:1313 arg1 OST 1325:1327 the only bacterial OST 1306:1327 the only bacterial OST 1306:1327 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 5 13 theme Desulfovibrio 993:1005 arg1 vulgaris 1007:1014 Desulfovibrio vulgaris 993:1014 Desulfovibrio vulgaris 993:1014 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 2 14 theme relative 542:549 arg1 position 533:540 the -2 position 526:540 the -2 position relative to the asparagine 526:567 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 7 15 theme Fc 1348:1349 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 15 theme Fc 1348:1349 arg1 domain 1351:1356 the Fc domain 1344:1356 the Fc domain of human immunoglobulin G at its native 'QYNST' sequon 1344:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 5 16 from desulfuricans 953:965 arg1 OSTs 882:885 the OSTs 878:885 the OSTs 878:885 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 6 17 theme multiple 1156:1163 arg1 targets 1165:1171 multiple targets 1156:1171 multiple targets 1156:1171 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 6 18 theme N-X-T 1140:1144 arg1 motifs 1146:1151 minimal N-X-T motifs 1132:1151 minimal N-X-T motifs in multiple targets 1132:1171 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 7 19 theme D. 1281:1282 arg1 PglB 1290:1293 D. gigas PglB 1281:1293 D. gigas PglB 1281:1293 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 5 20 theme Desulfovibrio 968:980 arg1 gigas 982:986 Desulfovibrio gigas 968:986 Desulfovibrio gigas 968:986 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 2 21 contain having 487:492 arg2 residue 515:521 a negatively charged residue 494:521 a negatively charged residue in the -2 position relative to the asparagine 494:567 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 2 21 contain having 487:492 arg1 those 481:485 those 481:485 those 481:485 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 7 22 from sequon 1406:1411 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 22 from sequon 1406:1411 arg1 domain 1351:1356 the Fc domain 1344:1356 the Fc domain of human immunoglobulin G at its native 'QYNST' sequon 1344:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 2 23 theme acceptor 415:422 arg1 restricted 453:462 restricted 453:462 restricted 453:462 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 2 23 theme acceptor 415:422 arg1 sites 424:428 the acceptor sites 411:428 the acceptor sites that it recognizes 411:447 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 7 24 theme notable 1262:1268 arg1 example 1270:1276 One notable example 1258:1276 One notable example 1258:1276 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 8 25 theme bacterial 1448:1456 arg1 OSTs 1458:1461 bacterial OSTs 1448:1461 bacterial OSTs 1448:1461 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 7 26 theme G 1382:1382 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 26 theme G 1382:1382 arg1 domain 1351:1356 the Fc domain 1344:1356 the Fc domain of human immunoglobulin G at its native 'QYNST' sequon 1344:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 27 dep D. 1281:1282 arg1 gigas 1284:1288 D. gigas 1281:1288 D. gigas PglB 1281:1293 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 4 28 theme trans-complementation 707:727 arg1 assay 729:733 an ectopic trans-complementation assay 696:733 an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli 696:797 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 1 29 theme target 319:324 arg1 proteins 326:333 target proteins 319:333 target proteins 319:333 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 5 30 from gigas 982:986 arg1 OSTs 882:885 the OSTs 878:885 the OSTs 878:885 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 1 31 theme central 118:124 arg1 oligosaccharyltransferase 208:232 the oligosaccharyltransferase 204:232 the oligosaccharyltransferase (OST) 204:238 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 1 31 theme central 118:124 arg1 enzyme 126:131 The central enzyme 114:131 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway 114:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 4 32 theme ectopic 699:705 arg1 assay 729:733 an ectopic trans-complementation assay 696:733 an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli 696:797 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 0 33 theme diverse 23:29 arg1 oligosaccharyltransferases 41:66 diverse bacterial oligosaccharyltransferases 23:66 sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences 11:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 4 34 theme OSTs 857:860 arg1 OSTs 857:860 the candidate OSTs 843:860 the candidate OSTs 843:860 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 4 34 theme OSTs 857:860 arg1 16 837:838 16 837:838 16 837:838 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 1 35 from residues 307:314 arg1 proteins 326:333 target proteins 319:333 target proteins 319:333 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 7 36 from domain 1351:1356 arg1 sequon 1406:1411 its native 'QYNST' sequon 1387:1411 its native 'QYNST' sequon 1387:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 4 37 theme in 816:817 arg1 activity 824:831 in vivo activity 816:831 in vivo activity for 16 of the candidate OSTs 816:860 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 7 38 theme immunoglobulin 1367:1380 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 3 39 theme acceptor-site 596:608 arg1 preferences 610:620 the acceptor-site preferences 592:620 the acceptor-site preferences for 23 homologs with natural sequence variation compared to CjPglB 592:687 Here, we investigated the acceptor-site preferences for 23 homologs with natural sequence variation compared to CjPglB. 26482295 4 40 theme CjPglB 739:744 arg1 function 746:753 CjPglB function 739:753 CjPglB function in glycosylation-competent Escherichia coli 739:797 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 5 41 theme Desulfovibrio 939:951 arg1 desulfuricans 953:965 Desulfovibrio desulfuricans 939:965 Desulfovibrio desulfuricans 939:965 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 2 42 theme charged 507:513 arg1 residue 515:521 a negatively charged residue 494:521 a negatively charged residue in the -2 position relative to the asparagine 494:567 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 2 43 from residue 515:521 arg1 position 533:540 the -2 position 526:540 the -2 position relative to the asparagine 526:567 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 6 44 theme minimal 1132:1138 arg1 motifs 1146:1151 minimal N-X-T motifs 1132:1151 minimal N-X-T motifs in multiple targets 1132:1171 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 0 45 theme bacterial 31:39 arg1 oligosaccharyltransferases 41:66 diverse bacterial oligosaccharyltransferases 23:66 sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences 11:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 8 46 theme acceptor-site 1490:1502 arg1 specificity 1504:1514 acceptor-site specificity 1490:1514 acceptor-site specificity 1490:1514 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 7 47 theme human 1361:1365 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 0 48 theme unique 73:78 arg1 preferences 101:111 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 7 49 theme QYNST 1399:1403 arg1 sequon 1406:1411 its native 'QYNST' sequon 1387:1411 its native 'QYNST' sequon 1387:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 5 50 from coli 906:909 arg1 OSTs 882:885 the OSTs 878:885 the OSTs 878:885 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 4 51 dep in 816:817 arg1 vivo 819:822 vivo 819:822 vivo 819:822 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 1 52 link asparagine-linked 161:177 arg1 pathway 193:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 6 53 dep unique 1191:1196 arg1 unknown 1206:1212 unknown 1206:1212 unknown 1206:1212 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 6 53 dep unique 1191:1196 arg1 as 1199:1200 as 1199:1200 as 1199:1200 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 1 54 theme Campylobacter 140:152 arg1 jejuni 154:159 Campylobacter jejuni 140:159 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 1 55 from enzyme 126:131 arg1 pathway 193:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 4 56 theme candidate 847:855 arg1 OSTs 857:860 the candidate OSTs 843:860 the candidate OSTs 843:860 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 8 57 theme OSTs 1458:1461 arg1 subset 1438:1443 a subset 1436:1443 a subset of bacterial OSTs 1436:1461 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 3 58 theme sequence 651:658 arg1 variation 660:668 natural sequence variation 643:668 natural sequence variation compared to CjPglB 643:687 Here, we investigated the acceptor-site preferences for 23 homologs with natural sequence variation compared to CjPglB. 26482295 1 59 theme jejuni 154:159 arg1 pathway 193:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 7 60 gly glycosylate 1332:1342 arg1 G 1382:1382 human immunoglobulin G 1361:1382 human immunoglobulin G at its native 'QYNST' sequon 1361:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 7 60 gly glycosylate 1332:1342 arg1 domain 1351:1356 the Fc domain 1344:1356 the Fc domain of human immunoglobulin G at its native 'QYNST' sequon 1344:1411 One notable example is D. gigas PglB, which was the only bacterial OST to glycosylate the Fc domain of human immunoglobulin G at its native 'QYNST' sequon. 26482295 5 61 theme relaxed 1041:1047 arg1 specificity 1049:1059 significantly relaxed specificity 1027:1059 significantly relaxed specificity towards the -2 position 1027:1083 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 5 62 from upsaliensis 926:936 arg1 OSTs 882:885 the OSTs 878:885 the OSTs 878:885 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 1 63 theme asparagine-linked 161:177 arg1 pathway 193:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 5 64 from vulgaris 1007:1014 arg1 OSTs 882:885 the OSTs 878:885 the OSTs 878:885 Interestingly, the OSTs from Campylobacter coli, Campylobacter upsaliensis, Desulfovibrio desulfuricans, Desulfovibrio gigas, and Desulfovibrio vulgaris, exhibited significantly relaxed specificity towards the -2 position compared to CjPglB. 26482295 1 65 theme preassembled 263:274 arg1 glycans 276:282 preassembled glycans 263:282 preassembled glycans 263:282 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 0 66 theme site 96:99 arg1 preferences 101:111 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 6 67 theme unique 1191:1196 arg1 rules 1215:1219 unique, as yet unknown, rules 1191:1219 unique, as yet unknown, rules governing acceptor-site preferences 1191:1255 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 26482295 0 68 dep sweeteners 11:20 arg1 oligosaccharyltransferases 41:66 diverse bacterial oligosaccharyltransferases 23:66 sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences 11:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 1 69 theme glycosylation 179:191 arg1 pathway 193:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 the Campylobacter jejuni asparagine-linked glycosylation pathway 136:199 The central enzyme in the Campylobacter jejuni asparagine-linked glycosylation pathway is the oligosaccharyltransferase (OST), PglB, which transfers preassembled glycans to specific asparagine residues in target proteins. 26482295 8 70 with toolbox 1556:1562 arg1 diversity 1605:1613 previously unavailable biocatalytic diversity 1569:1613 previously unavailable biocatalytic diversity 1569:1613 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 0 71 theme N-glycosylation 80:94 arg1 preferences 101:111 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 8 72 theme glycoengineering 1539:1554 arg1 toolbox 1556:1562 the glycoengineering toolbox 1535:1562 the glycoengineering toolbox with previously unavailable biocatalytic diversity 1535:1613 Overall, we find that a subset of bacterial OSTs follow their own rules for acceptor-site specificity, thereby expanding the glycoengineering toolbox with previously unavailable biocatalytic diversity. 26482295 3 73 theme natural 643:649 arg1 variation 660:668 natural sequence variation 643:668 natural sequence variation compared to CjPglB 643:687 Here, we investigated the acceptor-site preferences for 23 homologs with natural sequence variation compared to CjPglB. 26482295 0 74 with oligosaccharyltransferases 41:66 arg1 preferences 101:111 unique N-glycosylation site preferences 73:111 unique N-glycosylation site preferences 73:111 Substitute sweeteners: diverse bacterial oligosaccharyltransferases with unique N-glycosylation site preferences. 26482295 2 75 theme glycan 392:397 arg1 structures 399:408 many diverse glycan structures 379:408 many diverse glycan structures 379:408 While C. jejuni PglB (CjPglB) can transfer many diverse glycan structures, the acceptor sites that it recognizes are restricted predominantly to those having a negatively charged residue in the -2 position relative to the asparagine. 26482295 4 76 from function 746:753 arg1 coli 794:797 glycosylation-competent Escherichia coli 758:797 glycosylation-competent Escherichia coli 758:797 Using an ectopic trans-complementation assay for CjPglB function in glycosylation-competent Escherichia coli, we demonstrated in vivo activity for 16 of the candidate OSTs. 26482295 6 77 from motifs 1146:1151 arg1 targets 1165:1171 multiple targets 1156:1171 multiple targets 1156:1171 These enzymes glycosylated minimal N-X-T motifs in multiple targets and each followed unique, as yet unknown, rules governing acceptor-site preferences. 27641064 6 0 theme EGF 874:876 arg1 stimulation 878:888 EGF stimulation 874:888 EGF stimulation 874:888 Mechanistically, this N-glycosylation inhibited the response abilities upon EGF stimulation and EGFR dimerization. 27641064 7 1 theme N-glycosylation 942:956 arg1 formation 986:994 this N-glycosylation controlled the EGFR complex formation 937:994 this N-glycosylation controlled the EGFR complex formation 937:994 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 9 2 theme site-11 1348:1354 arg1 important 1386:1394 important 1386:1394 important 1386:1394 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 9 2 theme site-11 1348:1354 arg1 N-glycosylation 1356:1370 the site-11 N-glycosylation 1344:1370 the site-11 N-glycosylation on α5 1344:1376 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 0 3 theme complex 66:72 arg1 formation 74:82 EGFR-mediated complex formation 52:82 EGFR-mediated complex formation of integrin α5β1 52:99 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 2 4 theme EGFR 322:325 arg1 signaling 327:335 EGFR signaling 322:335 EGFR signaling in regulating cell proliferation 322:368 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 0 5 theme integrin 87:94 arg1 α5β1 96:99 integrin α5β1 87:99 integrin α5β1 87:99 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 9 6 theme regulatory 1467:1476 arg1 mechanism 1478:1486 a novel regulatory mechanism 1459:1486 a novel regulatory mechanism for crosstalks between integrins and EGFR 1459:1528 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 4 7 theme site-11 582:588 arg1 key 602:604 key 602:604 key 602:604 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 4 7 theme site-11 582:588 arg1 N-glycosylation 563:577 the N-glycosylation 559:577 the N-glycosylation of site-11 (Asn712) 559:597 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 7 8 theme cell 1137:1140 arg1 growth 1142:1147 cell growth 1137:1147 cell growth 1137:1147 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 6 9 theme response 850:857 arg1 abilities 859:867 the response abilities 846:867 the response abilities 846:867 Mechanistically, this N-glycosylation inhibited the response abilities upon EGF stimulation and EGFR dimerization. 27641064 8 10 theme EGFR-α5β1 1271:1279 arg1 formation 1281:1289 EGFR-α5β1 formation 1271:1289 EGFR-α5β1 formation 1271:1289 Moreover, the site-11 N-glycan exhibited a more branching structure compared with other sites, which may be required for EGFR-α5β1 formation. 27641064 5 11 theme cell 711:714 arg1 growth 716:721 cell growth 711:721 cell growth 711:721 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 7 12 theme site-11 1042:1048 arg1 loss 1034:1037 the loss 1030:1037 the loss of site-11 1030:1048 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 2 13 from effect 312:317 arg1 signaling 327:335 EGFR signaling 322:335 EGFR signaling in regulating cell proliferation 322:368 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 3 14 theme N-glycosylation 413:427 arg1 mechanisms 448:457 the underlying mechanisms 433:457 the underlying mechanisms of inhibition 433:471 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 3 14 theme N-glycosylation 413:427 arg1 importance 384:393 the importance 380:393 the importance of the individual N-glycosylation 380:427 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 0 15 theme α5β1 96:99 arg1 formation 74:82 EGFR-mediated complex formation 52:82 EGFR-mediated complex formation of integrin α5β1 52:99 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 2 16 theme integrin 254:261 arg1 S3-5,10-14 267:276 S3-5,10-14 267:276 S3-5,10-14 267:276 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 2 16 theme integrin 254:261 arg1 α5 263:264 integrin α5 254:264 integrin α5 (S3-5,10-14) 254:277 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 1 17 theme multiple 158:165 arg1 behaviors 172:180 multiple cell behaviors 158:180 multiple cell behaviors 158:180 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 27641064 7 18 theme cellular 1114:1121 arg1 signaling 1123:1131 cellular signaling 1114:1131 cellular signaling for cell growth 1114:1147 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 4 19 gly N-glycosylation 563:577 arg1 Asn712 591:596 Asn712 591:596 Asn712 591:596 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 4 19 gly N-glycosylation 563:577 arg1 site-11 582:588 site-11 582:588 site-11 (Asn712) 582:597 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 2 20 from N-glycosylations 215:230 arg1 S3-5,10-14 267:276 S3-5,10-14 267:276 S3-5,10-14 267:276 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 2 20 from N-glycosylations 215:230 arg1 α5 263:264 integrin α5 254:264 integrin α5 (S3-5,10-14) 254:277 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 1 21 theme cell 167:170 arg1 behaviors 172:180 multiple cell behaviors 158:180 multiple cell behaviors 158:180 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 27641064 2 22 theme cell 351:354 arg1 proliferation 356:368 cell proliferation 351:368 cell proliferation 351:368 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 0 23 theme α5 28:29 arg1 N-Glycosylation 0:14 N-Glycosylation 0:14 N-Glycosylation of integrin α5 0:29 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 0 23 theme α5 28:29 arg1 switch 41:46 a switch 39:46 a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4 39:107 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 5 24 dep wild-type 782:790 arg1 WT 793:794 WT 793:794 WT 793:794 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 5 25 theme other 662:666 arg1 sites 679:683 the other individual sites 658:683 the other individual sites 658:683 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 9 26 theme inhibitory 1404:1413 arg1 effect 1415:1420 its inhibitory effect 1400:1420 its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR 1400:1528 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 0 27 theme integrin 19:26 arg1 α5 28:29 integrin α5 19:29 integrin α5 19:29 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 5 28 theme individual 668:677 arg1 sites 679:683 the other individual sites 658:683 the other individual sites 658:683 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 9 29 theme novel 1461:1465 arg1 mechanism 1478:1486 a novel regulatory mechanism 1459:1486 a novel regulatory mechanism for crosstalks between integrins and EGFR 1459:1528 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 8 30 theme branching 1198:1206 arg1 structure 1208:1216 a more branching structure 1191:1216 a more branching structure 1191:1216 Moreover, the site-11 N-glycan exhibited a more branching structure compared with other sites, which may be required for EGFR-α5β1 formation. 27641064 2 31 gly N-glycosylations 215:230 arg1 domain 244:249 the calf domain 235:249 the calf domain 235:249 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 0 32 gly N-Glycosylation 0:14 arg1 α5 28:29 integrin α5 19:29 integrin α5 19:29 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 3 33 theme individual 402:411 arg1 N-glycosylation 413:427 the individual N-glycosylation 398:427 the individual N-glycosylation 398:427 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 4 34 from mutants 526:532 arg1 detail 537:542 detail 537:542 detail 537:542 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 4 35 theme S3-5,10-14 515:524 arg1 mutants 526:532 the S3-5,10-14 mutants 511:532 the S3-5,10-14 mutants in detail 511:542 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 1 36 gly N-Glycosylation 110:124 arg1 α5β1 138:141 integrin α5β1 129:141 integrin α5β1 129:141 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 27641064 1 36 gly N-Glycosylation 110:124 arg1 behaviors 172:180 multiple cell behaviors 158:180 multiple cell behaviors 158:180 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 27641064 9 37 from effect 1415:1420 arg1 signaling 1430:1438 EGFR signaling 1425:1438 EGFR signaling 1425:1438 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 3 38 theme underlying 437:446 arg1 mechanisms 448:457 the underlying mechanisms 433:457 the underlying mechanisms of inhibition 433:471 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 8 39 theme site-11 1164:1170 arg1 N-glycan 1172:1179 the site-11 N-glycan 1160:1179 the site-11 N-glycan 1160:1179 Moreover, the site-11 N-glycan exhibited a more branching structure compared with other sites, which may be required for EGFR-α5β1 formation. 27641064 5 40 theme site-11 642:648 arg1 restoration 627:637 The restoration 623:637 The restoration of site-11 623:648 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 9 41 theme EGFR 1425:1428 arg1 signaling 1430:1438 EGFR signaling 1425:1438 EGFR signaling 1425:1438 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 9 42 from N-glycosylation 1356:1370 arg1 α5 1375:1376 α5 1375:1376 α5 1375:1376 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 6 43 theme EGFR 894:897 arg1 dimerization 899:910 EGFR dimerization 894:910 EGFR dimerization 894:910 Mechanistically, this N-glycosylation inhibited the response abilities upon EGF stimulation and EGFR dimerization. 27641064 7 44 theme EGFR 973:976 arg1 complex 978:984 the EGFR complex 969:984 this N-glycosylation controlled the EGFR complex formation 937:994 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 8 45 theme other 1232:1236 arg1 sites 1238:1242 other sites 1232:1242 other sites 1232:1242 Moreover, the site-11 N-glycan exhibited a more branching structure compared with other sites, which may be required for EGFR-α5β1 formation. 27641064 7 46 theme integrin 1001:1008 arg1 α5β1 1010:1013 integrin α5β1 1001:1013 integrin α5β1 1001:1013 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 9 47 gly N-glycosylation 1356:1370 arg1 α5 1375:1376 α5 1375:1376 α5 1375:1376 Taken together, these data clearly demonstrate that the site-11 N-glycosylation on α5 is most important for its inhibitory effect on EGFR signaling, which may provide a novel regulatory mechanism for crosstalks between integrins and EGFR. 27641064 7 48 theme complex 978:984 arg1 formation 986:994 this N-glycosylation controlled the EGFR complex formation 937:994 this N-glycosylation controlled the EGFR complex formation 937:994 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 2 49 theme inhibitory 301:310 arg1 effect 312:317 its inhibitory effect 297:317 its inhibitory effect on EGFR signaling in regulating cell proliferation 297:368 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 2 50 theme domain 244:249 arg1 N-glycosylations 215:230 the N-glycosylations 211:230 the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) 211:277 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 2 50 theme domain 244:249 arg1 essential 283:291 essential 283:291 essential 283:291 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 7 51 theme controlled 958:967 arg1 formation 986:994 this N-glycosylation controlled the EGFR complex formation 937:994 this N-glycosylation controlled the EGFR complex formation 937:994 Interestingly, we found this N-glycosylation controlled the EGFR complex formation with integrin α5β1 or α6β4; i.e., the loss of site-11 switched EGFR-α5β1 to EGFR-α6β4, which is well known to promote cellular signaling for cell growth. 27641064 3 52 theme inhibition 462:471 arg1 mechanisms 448:457 the underlying mechanisms 433:457 the underlying mechanisms of inhibition 433:471 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 3 52 theme inhibition 462:471 arg1 importance 384:393 the importance 380:393 the importance of the individual N-glycosylation 380:427 However, the importance of the individual N-glycosylation and the underlying mechanisms of inhibition remain unclear. 27641064 0 53 theme EGFR-mediated 52:64 arg1 formation 74:82 EGFR-mediated complex formation 52:82 EGFR-mediated complex formation of integrin α5β1 52:99 N-Glycosylation of integrin α5 acts as a switch for EGFR-mediated complex formation of integrin α5β1 to α6β4. 27641064 4 54 theme cell 610:613 arg1 growth 615:620 cell growth 610:620 cell growth 610:620 Here, we characterize the S3-5,10-14 mutants in detail and found that the N-glycosylation of site-11 (Asn712) is key for cell growth. 27641064 2 55 theme calf 239:242 arg1 domain 244:249 the calf domain 235:249 the calf domain 235:249 We previously reported that the N-glycosylations of the calf domain on integrin α5 (S3-5,10-14) are essential for its inhibitory effect on EGFR signaling in regulating cell proliferation. 27641064 1 56 theme integrin 129:136 arg1 α5β1 138:141 integrin α5β1 129:141 integrin α5β1 129:141 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 27641064 5 57 theme EGFR 727:730 arg1 signaling 732:740 EGFR signaling 727:740 EGFR signaling 727:740 The restoration of site-11, unlike the other individual sites, significantly suppressed cell growth and EGFR signaling in a manner that was similar to that of wild-type (WT). 27641064 1 58 theme α5β1 138:141 arg1 N-Glycosylation 110:124 N-Glycosylation 110:124 N-Glycosylation of integrin α5β1 110:141 N-Glycosylation of integrin α5β1 is involved in multiple cell behaviors. 29164281 5 0 theme model 738:742 arg1 system 744:749 an animal model system 728:749 an animal model system 728:749 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 5 1 from limitation 766:775 arg1 humans 821:826 recently deceased humans 803:826 recently deceased humans 803:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 11 2 theme saliva 1638:1643 arg1 glycosylation 1645:1657 saliva glycosylation 1638:1657 saliva glycosylation 1638:1657 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 9 3 theme high 1307:1310 arg1 level 1312:1316 high level 1307:1316 high level 1307:1316 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 6 4 theme Rat 829:831 arg1 samples 840:846 Rat saliva samples 829:846 Rat saliva samples 829:846 Rat saliva samples were collected both before and after death. 29164281 4 5 theme various 540:546 arg1 glycoproteins 548:560 various glycoproteins 540:560 various glycoproteins that are highly affected by biochemical environment 540:612 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 8 6 theme native 988:993 arg1 glycans 995:1001 Released native glycans 979:1001 Released native glycans 979:1001 Released native glycans were purified and enriched by PGC-SPE. 29164281 5 7 theme saliva 637:642 arg1 N-glycans 644:652 saliva N-glycans 637:652 saliva N-glycans between live and dead rats 637:679 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 8 8 theme Released 979:986 arg1 glycans 995:1001 Released native glycans 979:1001 Released native glycans 979:1001 Released native glycans were purified and enriched by PGC-SPE. 29164281 11 9 theme glycosylation 1645:1657 arg1 changes 1627:1633 the post-mortem changes 1611:1633 the post-mortem changes of saliva glycosylation 1611:1657 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 7 10 theme PNGase 933:938 arg1 F 940:940 PNGase F 933:940 PNGase F 933:940 N-Glycans were enzymatically released by PNGase F without any glycoprotein extraction. 29164281 3 11 theme death 463:467 arg1 time 455:458 time 455:458 time of death 455:467 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 5 12 theme N-glycans 712:720 arg1 alteration 698:707 the alteration 694:707 the alteration of N-glycans 694:720 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 9 13 located detected 1295:1302 arg2 disaccharides 1276:1288 LacdiNAc disaccharides 1267:1288 LacdiNAc disaccharides 1267:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 13 located detected 1295:1302 arg1 level 1312:1316 high level 1307:1316 high level 1307:1316 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 13 located detected 1295:1302 arg2 N-glycans 1247:1255 non-sialylated N-glycans 1232:1255 non-sialylated N-glycans including LacdiNAc disaccharides 1232:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 11 14 theme first 1588:1592 arg1 this 1576:1579 this 1576:1579 this 1576:1579 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 11 14 theme first 1588:1592 arg1 study 1594:1598 the first study 1584:1598 the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications 1584:1693 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 1 15 theme crime 160:164 arg1 deaths 179:184 untimely deaths 170:184 untimely deaths in forensic science 170:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 1 15 theme crime 160:164 arg1 investigations 142:155 investigations 142:155 investigations of crime 142:164 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 0 16 theme saliva 37:42 arg1 N-glycosylation 44:58 saliva N-glycosylation 37:58 saliva N-glycosylation 37:58 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 29164281 2 17 theme estimation 240:249 arg1 methods 225:231 standard methods 216:231 standard methods of PMI estimation 216:249 However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. 29164281 9 18 from saliva 1217:1222 arg1 present 1184:1190 present 1184:1190 present 1184:1190 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 10 19 theme marker 1508:1513 arg1 N-glycans 1426:1434 14 N-glycans 1423:1434 14 N-glycans that significantly changed after death 1423:1473 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 10 19 theme marker 1508:1513 arg1 candidates 1515:1524 the potential marker candidates 1494:1524 the potential marker candidates for PMI estimation 1494:1543 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 4 20 gly glycoproteins 548:560 arg1 glycoproteins 548:560 various glycoproteins 540:560 various glycoproteins that are highly affected by biochemical environment 540:612 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 3 21 theme accurate 402:409 arg1 method 411:416 a more acceptable and accurate method 380:416 a more acceptable and accurate method 380:416 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 5 22 theme saliva 780:785 arg1 collection 787:796 saliva collection 780:796 saliva collection from recently deceased humans 780:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 10 23 theme potential 1498:1506 arg1 N-glycans 1426:1434 14 N-glycans 1423:1434 14 N-glycans that significantly changed after death 1423:1473 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 10 23 theme potential 1498:1506 arg1 candidates 1515:1524 the potential marker candidates 1494:1524 the potential marker candidates for PMI estimation 1494:1543 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 9 24 theme live 1208:1211 arg1 saliva 1217:1222 live rat saliva 1208:1222 live rat saliva 1208:1222 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 4 25 contain contains 531:538 arg1 one 478:480 one 478:480 one 478:480 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 4 25 contain contains 531:538 arg1 Saliva 470:475 Saliva 470:475 Saliva 470:475 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 4 25 contain contains 531:538 arg1 fluids 495:500 the vital fluids 485:500 the vital fluids encountered at crime scenes 485:528 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 4 25 contain contains 531:538 arg2 glycoproteins 548:560 various glycoproteins 540:560 various glycoproteins that are highly affected by biochemical environment 540:612 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 9 26 gly non-sialylated 1232:1245 arg1 N-glycans 1247:1255 non-sialylated N-glycans 1232:1255 non-sialylated N-glycans including LacdiNAc disaccharides 1232:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 26 gly non-sialylated 1232:1245 arg1 disaccharides 1276:1288 LacdiNAc disaccharides 1267:1288 LacdiNAc disaccharides 1267:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 3 27 theme PMI 354:356 arg1 markers 358:364 PMI markers 354:364 PMI markers 354:364 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 4 28 theme crime 517:521 arg1 scenes 523:528 crime scenes 517:528 crime scenes 517:528 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 9 29 theme rat 1213:1215 arg1 saliva 1217:1222 live rat saliva 1208:1222 live rat saliva 1208:1222 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 2 30 theme extenuating 276:286 arg1 circumstances 288:300 extenuating circumstances 276:300 extenuating circumstances 276:300 However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. 29164281 1 31 theme untimely 170:177 arg1 deaths 179:184 untimely deaths 170:184 untimely deaths in forensic science 170:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 3 32 theme markers 358:364 arg1 necessary 421:429 necessary 421:429 necessary 421:429 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 3 32 theme markers 358:364 arg1 panel 345:349 a panel 343:349 a panel of PMI markers obtained from a more acceptable and accurate method 343:416 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 7 33 theme glycoprotein 954:965 arg1 extraction 967:976 any glycoprotein extraction 950:976 any glycoprotein extraction 950:976 N-Glycans were enzymatically released by PNGase F without any glycoprotein extraction. 29164281 1 34 theme interval 105:112 arg1 part 133:136 a crucial part 123:136 a crucial part for investigations of crime and untimely deaths in forensic science 123:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 1 34 theme interval 105:112 arg1 estimation 79:88 The estimation 75:88 The estimation of post-mortem interval (PMI) 75:118 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 5 35 theme deceased 812:819 arg1 humans 821:826 recently deceased humans 803:826 recently deceased humans 803:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 3 36 theme acceptable 387:396 arg1 method 411:416 a more acceptable and accurate method 380:416 a more acceptable and accurate method 380:416 Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. 29164281 7 37 gly glycoprotein 954:965 arg1 glycoprotein 954:965 any glycoprotein extraction 950:976 any glycoprotein extraction 950:976 N-Glycans were enzymatically released by PNGase F without any glycoprotein extraction. 29164281 0 38 theme post-mortem 14:24 arg1 changes 26:32 post-mortem changes 14:32 post-mortem changes of saliva N-glycosylation 14:58 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 29164281 4 39 theme fluids 495:500 arg1 one 478:480 one 478:480 one 478:480 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 4 39 theme fluids 495:500 arg1 Saliva 470:475 Saliva 470:475 Saliva 470:475 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 4 39 theme fluids 495:500 arg1 fluids 495:500 the vital fluids 485:500 the vital fluids encountered at crime scenes 485:528 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 10 40 theme PMI 1530:1532 arg1 estimation 1534:1543 PMI estimation 1530:1543 PMI estimation 1530:1543 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 6 41 theme saliva 833:838 arg1 samples 840:846 Rat saliva samples 829:846 Rat saliva samples 829:846 Rat saliva samples were collected both before and after death. 29164281 0 42 theme changes 26:32 arg1 Monitoring 0:9 Monitoring 0:9 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 0:73 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 29164281 5 43 from humans 821:826 arg1 limitation 766:775 the limitation 762:775 the limitation of saliva collection from recently deceased humans 762:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 5 43 from humans 821:826 arg1 collection 787:796 saliva collection 780:796 saliva collection from recently deceased humans 780:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 5 44 theme collection 787:796 arg1 limitation 766:775 the limitation 762:775 the limitation of saliva collection from recently deceased humans 762:826 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 4 45 theme vital 489:493 arg1 fluids 495:500 the vital fluids 485:500 the vital fluids encountered at crime scenes 485:528 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 5 46 theme animal 731:736 arg1 system 744:749 an animal model system 728:749 an animal model system 728:749 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 9 47 theme nano 1120:1123 arg1 LC/MS 1125:1129 nano LC/MS 1120:1129 nano LC/MS 1120:1129 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 11 48 theme obvious 1665:1671 arg1 applications 1682:1693 obvious forensic applications 1665:1693 obvious forensic applications 1665:1693 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 0 49 theme N-glycosylation 44:58 arg1 changes 26:32 post-mortem changes 14:32 post-mortem changes of saliva N-glycosylation 14:58 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 29164281 5 50 theme live 662:665 arg1 rats 676:679 live and dead rats 662:679 live and dead rats 662:679 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 11 51 theme post-mortem 1615:1625 arg1 changes 1627:1633 the post-mortem changes 1611:1633 the post-mortem changes of saliva glycosylation 1611:1657 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 9 52 theme non-sialylated 1232:1245 arg1 N-glycans 1247:1255 non-sialylated N-glycans 1232:1255 non-sialylated N-glycans including LacdiNAc disaccharides 1232:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 52 theme non-sialylated 1232:1245 arg1 disaccharides 1276:1288 LacdiNAc disaccharides 1267:1288 LacdiNAc disaccharides 1267:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 10 53 theme statistical 1401:1411 arg1 analysis 1413:1420 statistical analysis 1401:1420 statistical analysis 1401:1420 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 9 54 theme LacdiNAc 1267:1274 arg1 disaccharides 1276:1288 LacdiNAc disaccharides 1267:1288 LacdiNAc disaccharides 1267:1288 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 2 55 theme PMI 236:238 arg1 estimation 240:249 PMI estimation 236:249 PMI estimation 236:249 However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. 29164281 1 56 from deaths 179:184 arg1 science 198:204 science 198:204 science 198:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 9 57 from abundance 1195:1203 arg1 present 1184:1190 present 1184:1190 present 1184:1190 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 1 58 theme post-mortem 93:103 arg1 PMI 115:117 PMI 115:117 PMI 115:117 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 1 58 theme post-mortem 93:103 arg1 interval 105:112 post-mortem interval 93:112 post-mortem interval (PMI) 93:118 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 5 59 theme dead 671:674 arg1 rats 676:679 live and dead rats 662:679 live and dead rats 662:679 Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. 29164281 9 60 theme Sialylated 1146:1155 arg1 N-glycans 1157:1165 tandem MS. Sialylated N-glycans 1135:1165 tandem MS. Sialylated N-glycans 1135:1165 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 61 theme tandem 1135:1140 arg1 N-glycans 1157:1165 tandem MS. Sialylated N-glycans 1135:1165 tandem MS. Sialylated N-glycans 1135:1165 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 2 62 theme environmental 309:321 arg1 factors 323:329 environmental factors 309:329 environmental factors 309:329 However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. 29164281 10 63 theme quantitative 1373:1384 arg1 comparison 1386:1395 quantitative comparison 1373:1395 quantitative comparison 1373:1395 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 10 64 theme in-depth 1343:1350 arg1 investigations 1352:1365 in-depth investigations 1343:1365 in-depth investigations using quantitative comparison and statistical analysis 1343:1420 Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. 29164281 9 65 located present 1184:1190 arg2 N-glycans 1157:1165 tandem MS. Sialylated N-glycans 1135:1165 tandem MS. Sialylated N-glycans 1135:1165 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 65 located present 1184:1190 arg1 saliva 1217:1222 live rat saliva 1208:1222 live rat saliva 1208:1222 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 65 located present 1184:1190 arg1 abundance 1195:1203 abundance 1195:1203 abundance 1195:1203 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 66 theme MS. 1142:1144 arg1 N-glycans 1157:1165 tandem MS. Sialylated N-glycans 1135:1165 tandem MS. Sialylated N-glycans 1135:1165 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 67 from present 1184:1190 arg1 saliva 1217:1222 live rat saliva 1208:1222 live rat saliva 1208:1222 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 67 from present 1184:1190 arg1 abundance 1195:1203 abundance 1195:1203 abundance 1195:1203 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 4 68 theme biochemical 590:600 arg1 environment 602:612 biochemical environment 590:612 biochemical environment 590:612 Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. 29164281 0 69 theme nano 63:66 arg1 LC/MS 68:72 nano LC/MS 63:72 nano LC/MS 63:72 Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. 29164281 11 70 theme forensic 1673:1680 arg1 applications 1682:1693 obvious forensic applications 1665:1693 obvious forensic applications 1665:1693 To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications. 29164281 2 71 theme standard 216:223 arg1 methods 225:231 standard methods 216:231 standard methods of PMI estimation 216:249 However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. 29164281 1 72 from investigations 142:155 arg1 science 198:204 science 198:204 science 198:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 1 73 theme crucial 125:131 arg1 part 133:136 a crucial part 123:136 a crucial part for investigations of crime and untimely deaths in forensic science 123:204 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 1 73 theme crucial 125:131 arg1 estimation 79:88 The estimation 75:88 The estimation of post-mortem interval (PMI) 75:118 The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. 29164281 9 74 attach present 1184:1190 arg2 N-glycans 1157:1165 tandem MS. Sialylated N-glycans 1135:1165 tandem MS. Sialylated N-glycans 1135:1165 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 74 attach present 1184:1190 arg1 saliva 1217:1222 live rat saliva 1208:1222 live rat saliva 1208:1222 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 29164281 9 74 attach present 1184:1190 arg1 abundance 1195:1203 abundance 1195:1203 abundance 1195:1203 About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. 28880909 4 0 theme medicinal 877:885 arg1 GONAL-f 895:901 its reference medicinal product GONAL-f 863:901 its reference medicinal product GONAL-f 863:901 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 12 1 theme Bemfola 2184:2190 arg1 variability 2169:2179 the greater bioactivity and higher batch-to-batch variability 2119:2179 the greater bioactivity and higher batch-to-batch variability of Bemfola 2119:2190 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 11 2 from differences 2001:2011 arg1 profile 2033:2039 the Asn52 glycan profile 2016:2039 the Asn52 glycan profile 2016:2039 These differences in the Asn52 glycan profile might potentially lead to differences in FSHR activation. 28880909 12 3 theme batch-to-batch 2154:2167 arg1 variability 2169:2179 the greater bioactivity and higher batch-to-batch variability 2119:2179 the greater bioactivity and higher batch-to-batch variability of Bemfola 2119:2190 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 12 4 dep batch-to-batch 2154:2167 arg1 higher 2147:2152 higher 2147:2152 higher 2147:2152 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 8 5 theme specific 1354:1361 arg1 IU/mg 1394:1398 14,522 IU/mg 1387:1398 14,522 IU/mg (105.6% of the nominal value) 1387:1428 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 5 theme specific 1354:1361 arg1 activity 1363:1370 the average specific activity 1342:1370 the average specific activity of Bemfola 1342:1381 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 2 6 theme biosimilar 297:306 arg1 versions 308:315 biosimilar versions 297:315 biosimilar versions of r-hFSH (follitropin alfa) 297:344 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 6 7 theme glycan 1139:1144 arg1 structures 1146:1155 bulkier glycan structures 1131:1155 bulkier glycan structures 1131:1155 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 10 8 theme % 1946:1946 arg1 proportion 1839:1848 a lower proportion 1831:1848 a lower proportion of bi-antennary structures [~53% vs ~77%] 1831:1890 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 8 theme % 1946:1946 arg1 proportion 1906:1915 a higher proportion 1897:1915 a higher proportion of tri-antennary [~41% vs ~23% 1897:1946 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 1 9 theme Recombinant 193:203 arg1 r-hFSH 241:246 r-hFSH 241:246 r-hFSH 241:246 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 1 9 theme Recombinant 193:203 arg1 hormone 232:238 Recombinant human follicle-stimulating hormone 193:238 Recombinant human follicle-stimulating hormone (r-hFSH) 193:247 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 3 10 theme active 577:582 arg1 component 584:592 active component 577:592 active component 577:592 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 8 11 theme average 1346:1352 arg1 IU/mg 1394:1398 14,522 IU/mg 1387:1398 14,522 IU/mg (105.6% of the nominal value) 1387:1428 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 11 theme average 1346:1352 arg1 activity 1363:1370 the average specific activity 1342:1370 the average specific activity of Bemfola 1342:1381 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 5 12 theme Asn52 1044:1048 arg1 glycosylation 1050:1062 Asn52 glycosylation 1044:1062 Asn52 glycosylation 1044:1062 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 10 13 located observed 1779:1786 arg1 Asn52 1791:1795 Asn52 1791:1795 Asn52 1791:1795 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 13 located observed 1779:1786 arg2 profile 1767:1773 A different glycan profile 1748:1773 A different glycan profile 1748:1773 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 13 located observed 1779:1786 arg1 Bemfola 1800:1806 Bemfola 1800:1806 Bemfola 1800:1806 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 1 14 theme follicle-stimulating 211:230 arg1 r-hFSH 241:246 r-hFSH 241:246 r-hFSH 241:246 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 1 14 theme follicle-stimulating 211:230 arg1 hormone 232:238 Recombinant human follicle-stimulating hormone 193:238 Recombinant human follicle-stimulating hormone (r-hFSH) 193:247 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 8 15 theme = 1550:1550 arg1 p 1548:1548 p = 0.0048 1548:1557 p = 0.0048 1548:1557 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 3 16 theme molecular 594:602 arg1 potency 636:642 potency 636:642 potency 636:642 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 3 16 theme molecular 594:602 arg1 impurities 621:630 impurities 621:630 impurities 621:630 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 3 16 theme molecular 594:602 arg1 characteristics 604:618 molecular characteristics 594:618 molecular characteristics 594:618 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 8 17 theme value 1423:1427 arg1 value 1423:1427 the nominal value 1411:1427 the nominal value 1411:1427 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 17 theme value 1423:1427 arg1 % 1406:1406 105.6% 1401:1406 105.6% of the nominal value 1401:1427 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 13 18 theme differences 2291:2301 arg1 relevance 2274:2282 The clinical relevance 2261:2282 The clinical relevance of the differences observed between GONAL-f and Bemfola 2261:2338 The clinical relevance of the differences observed between GONAL-f and Bemfola should be further investigated. 28880909 10 19 theme bi-antennary 1853:1864 arg1 structures 1866:1875 bi-antennary structures 1853:1875 bi-antennary structures 1853:1875 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 0 20 theme reference 154:162 arg1 GONAL-f 183:189 GONAL-f 183:189 GONAL-f 183:189 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 0 20 theme reference 154:162 arg1 product 174:180 its reference medicinal product 150:180 its reference medicinal product (GONAL-f) 150:190 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 8 21 dep IU/mg 1513:1517 arg1 % 1524:1524 97.3% 1520:1524 13,159 IU/mg; 97.3% of the nominal value 1506:1545 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 21 dep IU/mg 1513:1517 arg1 p 1548:1548 p = 0.0048 1548:1557 p = 0.0048 1548:1557 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 21 dep IU/mg 1513:1517 arg1 value 1541:1545 the nominal value 1529:1545 the nominal value 1529:1545 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 5 22 theme analysis 922:929 arg1 focus 908:912 The focus 904:912 The focus of this analysis 904:929 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 22 theme analysis 922:929 arg1 glycosylation 953:965 the site-specific glycosylation 935:965 the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH 935:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 9 23 theme batch-to-batch 1634:1647 arg1 variability 1649:1659 A higher batch-to-batch variability 1625:1659 A higher batch-to-batch variability 1625:1659 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 12 24 from differences 2227:2237 arg1 outcomes 2251:2258 clinical outcomes 2242:2258 clinical outcomes 2242:2258 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 2 25 theme reference 503:511 arg1 product 513:519 the reference product 499:519 the reference product 499:519 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 8 26 theme determined 1300:1309 arg1 potency 1311:1317 the determined potency 1296:1317 the determined potency 1296:1317 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 0 27 theme biosimilar 60:69 arg1 hormone 110:116 a biosimilar recombinant human follicle-stimulating hormone 58:116 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 3 28 dep differ 563:568 arg1 potency 636:642 potency 636:642 potency 636:642 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 3 28 dep differ 563:568 arg1 impurities 621:630 impurities 621:630 impurities 621:630 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 3 28 dep differ 563:568 arg1 characteristics 604:618 molecular characteristics 594:618 molecular characteristics 594:618 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 0 29 theme human 83:87 arg1 hormone 110:116 a biosimilar recombinant human follicle-stimulating hormone 58:116 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 5 30 gly glycosylation 953:965 arg1 α-subunit 997:1005 the α-subunit 993:1005 the α-subunit of FSH 993:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 30 gly glycosylation 953:965 arg1 Asn 982:984 asparagine (Asn) 52 970:988 asparagine (Asn) 52 970:988 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 30 gly glycosylation 953:965 arg2 Asn 982:984 asparagine (Asn) 52 970:988 asparagine (Asn) 52 970:988 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 8 31 theme average 1458:1464 arg1 activity 1475:1482 the average specific activity 1454:1482 the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048) 1454:1558 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 32 theme nominal 1327:1333 arg1 amount 1335:1340 the nominal amount 1323:1340 the nominal amount 1323:1340 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 0 33 theme hormone 110:116 arg1 product 118:124 a biosimilar recombinant human follicle-stimulating hormone product 58:124 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 0 33 theme hormone 110:116 arg1 Bemfola 127:133 Bemfola 127:133 Bemfola 127:133 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 5 34 theme receptor 1071:1078 arg1 activation/signalling 1087:1107 FSH receptor (FSHR) activation/signalling 1067:1107 FSH receptor (FSHR) activation/signalling 1067:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 11 35 theme FSHR 2082:2085 arg1 activation 2087:2096 FSHR activation 2082:2096 FSHR activation 2082:2096 These differences in the Asn52 glycan profile might potentially lead to differences in FSHR activation. 28880909 0 36 theme recombinant 71:81 arg1 hormone 110:116 a biosimilar recombinant human follicle-stimulating hormone 58:116 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 4 37 theme glycosylation 735:747 arg1 profile 749:755 the site-specific glycosylation profile 717:755 the site-specific glycosylation profile 717:755 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 0 38 theme In-vivo 0:6 arg1 activity 19:26 In-vivo biological activity 0:26 In-vivo biological activity 0:26 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 9 39 theme variation 1722:1730 arg1 coefficient 1707:1717 coefficient 1707:1717 coefficient of variation: 8.3% vs 5.8% 1707:1744 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 5 40 theme asparagine 970:979 arg1 Asn 982:984 asparagine (Asn) 52 970:988 asparagine (Asn) 52 970:988 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 2 41 theme manufacturing 413:425 arg1 process 427:433 manufacturing process 413:433 manufacturing process 413:433 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 0 42 theme glycosylation 32:44 arg1 analysis 46:53 glycosylation analysis 32:53 glycosylation analysis 32:53 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 10 43 dep observed 1779:1786 arg1 % 1983:1983 [~5% 1980:1983 [~5% 1980:1983 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 43 dep observed 1779:1786 arg1 % 1990:1990 <1% 1988:1990 <1% 1988:1990 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 4 44 theme follitropin 840:850 arg1 alfa 852:855 a biosimilar follitropin alfa 827:855 a biosimilar follitropin alfa 827:855 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 4 44 theme follitropin 840:850 arg1 Bemfola 818:824 Bemfola 818:824 Bemfola 818:824 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 10 45 theme tri-antennary 1920:1932 arg1 % 1938:1938 tri-antennary [~41% 1920:1938 tri-antennary [~41% 1920:1938 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 12 46 theme clinical 2242:2249 arg1 outcomes 2251:2258 clinical outcomes 2242:2258 clinical outcomes 2242:2258 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 4 47 theme bioactivity 803:813 arg1 profile 749:755 the site-specific glycosylation profile 717:755 the site-specific glycosylation profile 717:755 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 4 47 theme bioactivity 803:813 arg1 variability 776:786 batch-to-batch variability 761:786 batch-to-batch variability 761:786 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 12 48 dep bioactivity 2131:2141 arg1 greater 2123:2129 greater 2123:2129 greater 2123:2129 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 7 49 theme 13,636 1257:1262 arg1 activity 1216:1223 The nominal specific activity 1195:1223 The nominal specific activity for both Bemfola and GONAL-f 1195:1252 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 7 49 theme 13,636 1257:1262 arg1 IU/mg 1264:1268 13,636 IU/mg 1257:1268 13,636 IU/mg 1257:1268 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 8 50 theme nominal 1533:1539 arg1 value 1541:1545 the nominal value 1529:1545 the nominal value 1529:1545 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 10 51 theme different 1750:1758 arg1 profile 1767:1773 A different glycan profile 1748:1773 A different glycan profile 1748:1773 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 4 52 theme Bemfola 818:824 arg1 bioactivity 803:813 the in-vivo bioactivity 791:813 the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa 791:855 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 7 53 theme nominal 1199:1205 arg1 IU/mg 1264:1268 13,636 IU/mg 1257:1268 13,636 IU/mg 1257:1268 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 7 53 theme nominal 1199:1205 arg1 activity 1216:1223 The nominal specific activity 1195:1223 The nominal specific activity for both Bemfola and GONAL-f 1195:1252 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 4 54 theme reference 867:875 arg1 GONAL-f 895:901 its reference medicinal product GONAL-f 863:901 its reference medicinal product GONAL-f 863:901 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 10 55 theme % 1938:1938 arg1 proportion 1839:1848 a lower proportion 1831:1848 a lower proportion of bi-antennary structures [~53% vs ~77%] 1831:1890 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 55 theme % 1938:1938 arg1 proportion 1906:1915 a higher proportion 1897:1915 a higher proportion of tri-antennary [~41% vs ~23% 1897:1946 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 6 56 contain had 1127:1129 arg2 sialylation 1169:1179 greater sialylation 1161:1179 greater sialylation 1161:1179 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 6 56 contain had 1127:1129 arg1 Bemfola 1119:1125 Bemfola 1119:1125 Bemfola 1119:1125 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 6 56 contain had 1127:1129 arg2 structures 1146:1155 bulkier glycan structures 1131:1155 bulkier glycan structures 1131:1155 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 3 57 theme clinical 679:686 arg1 outcome 688:694 clinical outcome 679:694 clinical outcome 679:694 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 9 58 dep coefficient 1707:1717 arg1 % 1736:1736 8.3% 1733:1736 8.3% 1733:1736 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 9 58 dep coefficient 1707:1717 arg1 % 1744:1744 5.8% 1741:1744 5.8% 1741:1744 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 5 59 from Asn 982:984 arg1 focus 908:912 The focus 904:912 The focus of this analysis 904:929 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 59 from Asn 982:984 arg1 glycosylation 953:965 the site-specific glycosylation 935:965 the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH 935:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 1 60 theme fertility 267:275 arg1 treatment 277:285 fertility treatment 267:285 fertility treatment 267:285 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 4 61 theme product 887:893 arg1 GONAL-f 895:901 its reference medicinal product GONAL-f 863:901 its reference medicinal product GONAL-f 863:901 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 6 62 theme greater 1161:1167 arg1 sialylation 1169:1179 greater sialylation 1161:1179 greater sialylation 1161:1179 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 10 63 theme tetra-antennary 1953:1967 arg1 structures 1969:1978 tetra-antennary structures 1953:1978 tetra-antennary structures 1953:1978 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 7 64 theme specific 1207:1214 arg1 IU/mg 1264:1268 13,636 IU/mg 1257:1268 13,636 IU/mg 1257:1268 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 7 64 theme specific 1207:1214 arg1 activity 1216:1223 The nominal specific activity 1195:1223 The nominal specific activity for both Bemfola and GONAL-f 1195:1252 The nominal specific activity for both Bemfola and GONAL-f is 13,636 IU/mg. 28880909 2 65 theme follitropin 328:338 arg1 r-hFSH 320:325 r-hFSH 320:325 r-hFSH (follitropin alfa) 320:344 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 2 65 theme follitropin 328:338 arg1 alfa 340:343 follitropin alfa 328:343 follitropin alfa 328:343 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 3 66 dep component 584:592 arg1 e.g. 571:574 e.g. 571:574 e.g. 571:574 This evaluation should focus on how they differ (e.g., active component molecular characteristics, impurities and potency), as this could be associated with clinical outcome. 28880909 6 67 theme bulkier 1131:1137 arg1 structures 1146:1155 bulkier glycan structures 1131:1155 bulkier glycan structures 1131:1155 Overall, Bemfola had bulkier glycan structures and greater sialylation than GONAL-f. 28880909 10 68 theme lower 1833:1837 arg1 proportion 1839:1848 a lower proportion 1831:1848 a lower proportion of bi-antennary structures [~53% vs ~77%] 1831:1890 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 2 69 theme r-hFSH 320:325 arg1 versions 308:315 biosimilar versions 297:315 biosimilar versions of r-hFSH (follitropin alfa) 297:344 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 1 70 theme human 205:209 arg1 r-hFSH 241:246 r-hFSH 241:246 r-hFSH 241:246 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 1 70 theme human 205:209 arg1 hormone 232:238 Recombinant human follicle-stimulating hormone 193:238 Recombinant human follicle-stimulating hormone (r-hFSH) 193:247 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 8 71 theme nominal 1415:1421 arg1 value 1423:1427 the nominal value 1411:1427 the nominal value 1411:1427 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 5 72 theme glycosylation 1050:1062 arg1 role 1036:1039 the pivotal role 1024:1039 the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling 1024:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 73 gly glycosylation 1050:1062 arg1 FSHR 1081:1084 FSHR 1081:1084 FSHR 1081:1084 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 73 gly glycosylation 1050:1062 arg1 receptor 1071:1078 FSH receptor 1067:1078 FSH receptor (FSHR) activation/signalling 1067:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 10 74 theme structures 1866:1875 arg1 % 1881:1881 [~53% 1877:1881 [~53% 1877:1881 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 74 theme structures 1866:1875 arg1 structures 1866:1875 bi-antennary structures 1853:1875 bi-antennary structures 1853:1875 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 0 75 theme medicinal 164:172 arg1 GONAL-f 183:189 GONAL-f 183:189 GONAL-f 183:189 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 0 75 theme medicinal 164:172 arg1 product 174:180 its reference medicinal product 150:180 its reference medicinal product (GONAL-f) 150:190 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 5 76 theme FSH 1067:1069 arg1 FSHR 1081:1084 FSHR 1081:1084 FSHR 1081:1084 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 76 theme FSH 1067:1069 arg1 receptor 1071:1078 FSH receptor 1067:1078 FSH receptor (FSHR) activation/signalling 1067:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 9 77 theme higher 1627:1632 arg1 variability 1649:1659 A higher batch-to-batch variability 1625:1659 A higher batch-to-batch variability 1625:1659 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 9 78 dep Bemfola 1683:1689 arg1 coefficient 1707:1717 coefficient 1707:1717 coefficient of variation: 8.3% vs 5.8% 1707:1744 A higher batch-to-batch variability was also observed for Bemfola versus GONAL-f (coefficient of variation: 8.3% vs 5.8%). 28880909 5 79 theme pivotal 1028:1034 arg1 role 1036:1039 the pivotal role 1024:1039 the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling 1024:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 11 80 from differences 2067:2077 arg1 activation 2087:2096 FSHR activation 2082:2096 FSHR activation 2082:2096 These differences in the Asn52 glycan profile might potentially lead to differences in FSHR activation. 28880909 12 81 theme bioactivity 2131:2141 arg1 variability 2169:2179 the greater bioactivity and higher batch-to-batch variability 2119:2179 the greater bioactivity and higher batch-to-batch variability of Bemfola 2119:2190 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 28880909 5 82 from role 1036:1039 arg1 activation/signalling 1087:1107 FSH receptor (FSHR) activation/signalling 1067:1107 FSH receptor (FSHR) activation/signalling 1067:1107 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 8 83 dep IU/mg 1394:1398 arg1 value 1423:1427 the nominal value 1411:1427 the nominal value 1411:1427 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 83 dep IU/mg 1394:1398 arg1 % 1406:1406 105.6% 1401:1406 105.6% of the nominal value 1401:1427 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 83 dep IU/mg 1394:1398 arg1 greater 1441:1447 greater 1441:1447 greater 1441:1447 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 11 84 theme glycan 2026:2031 arg1 profile 2033:2039 the Asn52 glycan profile 2016:2039 the Asn52 glycan profile 2016:2039 These differences in the Asn52 glycan profile might potentially lead to differences in FSHR activation. 28880909 4 85 gly glycosylation 735:747 arg1 bioactivity 803:813 the in-vivo bioactivity 791:813 the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa 791:855 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 0 86 theme follicle-stimulating 89:108 arg1 hormone 110:116 a biosimilar recombinant human follicle-stimulating hormone 58:116 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 11 87 theme Asn52 2020:2024 arg1 profile 2033:2039 the Asn52 glycan profile 2016:2039 the Asn52 glycan profile 2016:2039 These differences in the Asn52 glycan profile might potentially lead to differences in FSHR activation. 28880909 8 88 theme specific 1466:1473 arg1 activity 1475:1482 the average specific activity 1454:1482 the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048) 1454:1558 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 0 89 theme product 118:124 arg1 activity 19:26 In-vivo biological activity 0:26 In-vivo biological activity 0:26 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 0 89 theme product 118:124 arg1 analysis 46:53 glycosylation analysis 32:53 glycosylation analysis 32:53 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 2 90 with comparison 483:492 arg1 product 513:519 the reference product 499:519 the reference product 499:519 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 4 91 theme batch-to-batch 761:774 arg1 variability 776:786 batch-to-batch variability 761:786 batch-to-batch variability 761:786 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 13 92 theme clinical 2265:2272 arg1 relevance 2274:2282 The clinical relevance 2261:2282 The clinical relevance of the differences observed between GONAL-f and Bemfola 2261:2338 The clinical relevance of the differences observed between GONAL-f and Bemfola should be further investigated. 28880909 10 93 dep proportion 1839:1848 arg1 % 1881:1881 [~53% 1877:1881 [~53% 1877:1881 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 93 dep proportion 1839:1848 arg1 % 1889:1889 ~77% 1886:1889 ~77% 1886:1889 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 93 dep proportion 1839:1848 arg1 structures 1866:1875 bi-antennary structures 1853:1875 bi-antennary structures 1853:1875 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 2 94 theme structural 387:396 arg1 complexity 398:407 their structural complexity 381:407 their structural complexity 381:407 Although biosimilar versions of r-hFSH (follitropin alfa) are currently on the market, given their structural complexity and manufacturing process, it is important to thoroughly evaluate them in comparison with the reference product. 28880909 0 95 theme biological 8:17 arg1 activity 19:26 In-vivo biological activity 0:26 In-vivo biological activity 0:26 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 5 96 theme site-specific 939:951 arg1 focus 908:912 The focus 904:912 The focus of this analysis 904:929 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 96 theme site-specific 939:951 arg1 glycosylation 953:965 the site-specific glycosylation 935:965 the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH 935:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 8 97 dep product 1610:1616 arg1 label 1618:1622 label 1618:1622 label 1618:1622 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 4 98 theme site-specific 721:733 arg1 profile 749:755 the site-specific glycosylation profile 717:755 the site-specific glycosylation profile 717:755 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 10 99 dep GONAL-f 1822:1828 arg1 proportion 1839:1848 a lower proportion 1831:1848 a lower proportion of bi-antennary structures [~53% vs ~77%] 1831:1890 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 10 99 dep GONAL-f 1822:1828 arg1 proportion 1906:1915 a higher proportion 1897:1915 a higher proportion of tri-antennary [~41% vs ~23% 1897:1946 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 5 100 theme FSH 1010:1012 arg1 α-subunit 997:1005 the α-subunit 993:1005 the α-subunit of FSH 993:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 8 101 theme Bemfola 1375:1381 arg1 IU/mg 1394:1398 14,522 IU/mg 1387:1398 14,522 IU/mg (105.6% of the nominal value) 1387:1428 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 101 theme Bemfola 1375:1381 arg1 activity 1363:1370 the average specific activity 1342:1370 the average specific activity of Bemfola 1342:1381 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 1 102 used used 259:262 arg2 r-hFSH 241:246 r-hFSH 241:246 r-hFSH 241:246 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 1 102 used used 259:262 arg2 hormone 232:238 Recombinant human follicle-stimulating hormone 193:238 Recombinant human follicle-stimulating hormone (r-hFSH) 193:247 Recombinant human follicle-stimulating hormone (r-hFSH) is widely used in fertility treatment. 28880909 4 103 theme biosimilar 829:838 arg1 alfa 852:855 a biosimilar follitropin alfa 827:855 a biosimilar follitropin alfa 827:855 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 4 103 theme biosimilar 829:838 arg1 Bemfola 818:824 Bemfola 818:824 Bemfola 818:824 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 0 104 gly glycosylation 32:44 arg1 product 118:124 a biosimilar recombinant human follicle-stimulating hormone product 58:124 a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) 58:134 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 0 104 gly glycosylation 32:44 arg1 Bemfola 127:133 Bemfola 127:133 Bemfola 127:133 In-vivo biological activity and glycosylation analysis of a biosimilar recombinant human follicle-stimulating hormone product (Bemfola) compared with its reference medicinal product (GONAL-f). 28880909 8 105 theme value 1541:1545 arg1 % 1524:1524 97.3% 1520:1524 13,159 IU/mg; 97.3% of the nominal value 1506:1545 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 8 105 theme value 1541:1545 arg1 value 1541:1545 the nominal value 1529:1545 the nominal value 1529:1545 Taking into account both the determined potency and the nominal amount the average specific activity of Bemfola was 14,522 IU/mg (105.6% of the nominal value), which was greater than the average specific activity observed for GONAL-f (13,159 IU/mg; 97.3% of the nominal value; p = 0.0048), although this was within the range stated in the product label. 28880909 10 106 theme glycan 1760:1765 arg1 profile 1767:1773 A different glycan profile 1748:1773 A different glycan profile 1748:1773 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 4 107 theme in-vivo 795:801 arg1 bioactivity 803:813 the in-vivo bioactivity 791:813 the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa 791:855 This study compared the site-specific glycosylation profile and batch-to-batch variability of the in-vivo bioactivity of Bemfola, a biosimilar follitropin alfa, with its reference medicinal product GONAL-f. 28880909 5 108 theme α-subunit 997:1005 arg1 focus 908:912 The focus 904:912 The focus of this analysis 904:929 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 5 108 theme α-subunit 997:1005 arg1 glycosylation 953:965 the site-specific glycosylation 935:965 the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH 935:1012 The focus of this analysis was the site-specific glycosylation at asparagine (Asn) 52 of the α-subunit of FSH, owing to the pivotal role of Asn52 glycosylation in FSH receptor (FSHR) activation/signalling. 28880909 10 109 theme higher 1899:1904 arg1 proportion 1906:1915 a higher proportion 1897:1915 a higher proportion of tri-antennary [~41% vs ~23% 1897:1946 A different glycan profile was observed at Asn52 in Bemfola compared with GONAL-f (a lower proportion of bi-antennary structures [~53% vs ~77%], and a higher proportion of tri-antennary [~41% vs ~23%] and tetra-antennary structures [~5% vs <1%]). 28880909 12 110 theme reported 2218:2225 arg1 differences 2227:2237 the reported differences 2214:2237 the reported differences in clinical outcomes 2214:2258 This, together with the greater bioactivity and higher batch-to-batch variability of Bemfola, could partly explain the reported differences in clinical outcomes. 27235585 12 0 from structure 1966:1974 arg1 other 1941:1945 other 1941:1945 other 1941:1945 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 3 1 theme level 645:649 arg1 understanding 651:663 the molecular level understanding 631:663 the molecular level understanding of carbonate binding site 631:689 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 9 2 theme straightforward 1353:1367 arg1 manner 1369:1374 a straightforward manner 1351:1374 a straightforward manner rather than more parameters required for full length protein 1351:1435 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 8 3 contain have 1141:1144 arg2 effect 1149:1154 an effect 1146:1154 an effect 1146:1154 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 8 3 contain have 1141:1144 arg1 Glycosylation 1116:1128 Glycosylation 1116:1128 Glycosylation 1116:1128 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 12 4 from other 1941:1945 arg1 structure 1966:1974 the transferrin structure 1950:1974 the transferrin structure 1950:1974 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 3 5 theme binding 678:684 arg1 site 686:689 carbonate binding site 668:689 carbonate binding site 668:689 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 1 6 theme iron 167:170 arg1 anion 188:192 a carbonate anion 176:192 a carbonate anion to form a Fe-CO3-Tf ternary complex 176:228 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 1 6 theme iron 167:170 arg1 binding 156:162 the highly cooperative binding 133:162 the highly cooperative binding of iron 133:170 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 8 7 theme transferrin 1202:1212 arg1 structure 1214:1222 transferrin structure 1202:1222 transferrin structure 1202:1222 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 2 8 theme residues 271:278 arg1 layout 244:249 the layout 240:249 the layout of the binding site residues 240:278 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 12 9 theme lobes 1927:1931 arg1 importance 1863:1872 the importance 1859:1872 the importance of the interlobe communication 1859:1903 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 12 9 theme lobes 1927:1931 arg1 impact 1913:1918 the impact 1909:1918 the impact of the lobes on each other in the transferrin structure 1909:1974 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 10 10 theme full 1520:1523 arg1 transferrin 1532:1542 full length transferrin 1520:1542 full length transferrin 1520:1542 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 2 11 theme binding 258:264 arg1 residues 271:278 the binding site residues 254:278 the binding site residues 254:278 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 9 12 theme full 1417:1420 arg1 protein 1429:1435 full length protein 1417:1435 full length protein 1417:1435 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 12 13 theme communication 1891:1903 arg1 importance 1863:1872 the importance 1859:1872 the importance of the interlobe communication 1859:1903 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 12 13 theme communication 1891:1903 arg1 impact 1913:1918 the impact 1909:1918 the impact of the lobes on each other in the transferrin structure 1909:1974 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 2 14 theme C-lobe 512:517 arg1 site 527:530 the C-lobe binding site 508:530 the C-lobe binding site 508:530 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 8 15 theme binding 1177:1183 arg1 site 1185:1188 the binding site residue and transferrin structure 1173:1222 the binding site residue and transferrin structure 1173:1222 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 5 16 gly glycoprotein 857:868 arg1 transferrin 840:850 native human transferrin 827:850 native human transferrin 827:850 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 5 16 gly glycoprotein 857:868 arg1 glycoprotein 857:868 a glycoprotein 855:868 a glycoprotein that two N-linked complex glycan chains located in the C-lobe 855:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 5 17 theme complex 888:894 arg1 chains 903:908 two N-linked complex glycan chains 875:908 two N-linked complex glycan chains located in the C-lobe 875:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 9 18 theme other 1232:1236 arg1 hand 1238:1241 the other hand 1228:1241 the other hand 1228:1241 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 3 19 theme molecular 569:577 arg1 simulation 587:596 molecular dynamic simulation 569:596 molecular dynamic simulation 569:596 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 11 20 theme significant 1720:1730 arg1 variation 1743:1751 a significant structural variation 1718:1751 a significant structural variation 1718:1751 Results revealed that separation in C-lobe caused a significant structural variation in comparison to N-lobe. 27235585 0 21 theme molecular 60:68 arg1 analysis 89:96 A molecular dynamic simulation analysis 58:96 A molecular dynamic simulation analysis 58:96 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 6 22 theme dynamic 959:965 arg1 simulation 967:976 the molecular dynamic simulation 945:976 the molecular dynamic simulation for simplifying 945:992 Usually, in the molecular dynamic simulation for simplifying, glycan is removed from the protein structure. 27235585 0 23 theme simulation 78:87 arg1 analysis 89:96 A molecular dynamic simulation analysis 58:96 A molecular dynamic simulation analysis 58:96 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 3 24 from interactions 701:712 arg1 lobe 722:725 each lobe 717:725 each lobe 717:725 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 2 25 dep N-lobe 339:344 arg1 contrast 327:334 contrast 327:334 contrast 327:334 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 10 26 theme molecular 1582:1590 arg1 simulation 1600:1609 the molecular dynamic simulation 1578:1609 the molecular dynamic simulation 1578:1609 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 2 27 theme C-lobe 347:352 arg1 site 362:365 C-lobe binding site 347:365 C-lobe binding site of the transferrin structure 347:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 2 27 theme C-lobe 347:352 arg1 structure 386:394 the transferrin structure 370:394 the transferrin structure 370:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 6 28 theme protein 1022:1028 arg1 structure 1030:1038 the protein structure 1018:1038 the protein structure 1018:1038 Usually, in the molecular dynamic simulation for simplifying, glycan is removed from the protein structure. 27235585 2 29 theme structure 386:394 arg1 site 362:365 C-lobe binding site 347:365 C-lobe binding site of the transferrin structure 347:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 2 29 theme structure 386:394 arg1 structure 386:394 the transferrin structure 370:394 the transferrin structure 370:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 8 30 dep site 1185:1188 arg1 residue 1190:1196 residue 1190:1196 residue 1190:1196 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 8 30 dep site 1185:1188 arg1 structure 1214:1222 transferrin structure 1202:1222 transferrin structure 1202:1222 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 2 31 theme little 428:433 arg1 research 435:442 little research 428:442 little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site 428:530 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 9 32 theme more 1388:1391 arg1 parameters 1393:1402 more parameters 1388:1402 more parameters required for full length protein 1388:1435 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 10 33 dep performed 1615:1623 arg1 are 1473:1475 are 1473:1475 are differences between the separated lobe and full length transferrin 1473:1542 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 10 34 theme structural 1645:1654 arg1 variations 1656:1665 such structural variations 1640:1665 such structural variations 1640:1665 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 7 35 theme transferrin 1093:1103 arg1 structure 1105:1113 the transferrin structure 1089:1113 the transferrin structure 1089:1113 Here, we explore the effect of glycosylation on the transferrin structure. 27235585 0 36 gly glycosylation 14:26 arg1 transferrin 35:45 the transferrin structure 31:55 the transferrin structure 31:55 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 3 37 theme molecular 635:643 arg1 understanding 651:663 the molecular level understanding 631:663 the molecular level understanding of carbonate binding site 631:689 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 5 38 from C-lobe 925:930 arg1 located 910:916 located 910:916 located 910:916 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 3 39 from access 619:624 arg1 lobe 722:725 each lobe 717:725 each lobe 717:725 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 4 40 theme transferrin 774:784 arg1 structure 786:794 transferrin structure 774:794 transferrin structure 774:794 Residues responsible for carbonate binding of transferrin structure were pointed out. 27235585 5 41 theme human 834:838 arg1 transferrin 840:850 native human transferrin 827:850 native human transferrin 827:850 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 5 41 theme human 834:838 arg1 glycoprotein 857:868 a glycoprotein 855:868 a glycoprotein that two N-linked complex glycan chains located in the C-lobe 855:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 1 42 theme ternary 214:220 arg1 complex 222:228 a Fe-CO3-Tf ternary complex 202:228 a Fe-CO3-Tf ternary complex 202:228 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 3 43 theme carbonate 668:676 arg1 site 686:689 carbonate binding site 668:689 carbonate binding site 668:689 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 7 44 theme glycosylation 1072:1084 arg1 effect 1062:1067 the effect 1058:1067 the effect of glycosylation on the transferrin structure 1058:1113 Here, we explore the effect of glycosylation on the transferrin structure. 27235585 6 45 attach removed 1005:1011 arg2 glycan 995:1000 glycan 995:1000 glycan 995:1000 Usually, in the molecular dynamic simulation for simplifying, glycan is removed from the protein structure. 27235585 6 45 attach removed 1005:1011 arg1 structure 1030:1038 the protein structure 1018:1038 the protein structure 1018:1038 Usually, in the molecular dynamic simulation for simplifying, glycan is removed from the protein structure. 27235585 10 46 theme separated 1501:1509 arg1 lobe 1511:1514 the separated lobe 1497:1514 the separated lobe 1497:1514 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 5 47 theme native 827:832 arg1 transferrin 840:850 native human transferrin 827:850 native human transferrin 827:850 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 5 47 theme native 827:832 arg1 glycoprotein 857:868 a glycoprotein 855:868 a glycoprotein that two N-linked complex glycan chains located in the C-lobe 855:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 0 48 theme transferrin 35:45 arg1 structure 47:55 the transferrin structure 31:55 the transferrin structure 31:55 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 8 49 theme site 1185:1188 arg1 layout 1163:1168 the layout 1159:1168 the layout of the binding site residue and transferrin structure 1159:1222 Glycosylation appears to have an effect on the layout of the binding site residue and transferrin structure. 27235585 3 50 theme site 686:689 arg1 understanding 651:663 the molecular level understanding 631:663 the molecular level understanding of carbonate binding site 631:689 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 10 51 theme length 1525:1530 arg1 transferrin 1532:1542 full length transferrin 1520:1542 full length transferrin 1520:1542 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 2 52 theme site 266:269 arg1 residues 271:278 the binding site residues 254:278 the binding site residues 254:278 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 1 53 theme carbonate 178:186 arg1 anion 188:192 a carbonate anion 176:192 a carbonate anion to form a Fe-CO3-Tf ternary complex 176:228 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 2 54 theme binding 519:525 arg1 site 527:530 the C-lobe binding site 508:530 the C-lobe binding site 508:530 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 7 55 gly glycosylation 1072:1084 arg1 transferrin 1093:1103 the transferrin structure 1089:1113 the transferrin structure 1089:1113 Here, we explore the effect of glycosylation on the transferrin structure. 27235585 10 56 theme comparative 1554:1564 arg1 analysis 1566:1573 a comparative analysis 1552:1573 a comparative analysis by the molecular dynamic simulation 1552:1609 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 3 57 theme present 555:561 arg1 work 563:566 the present work 551:566 the present work 551:566 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 9 58 theme length 1422:1427 arg1 protein 1429:1435 full length protein 1417:1435 full length protein 1417:1435 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 12 59 theme interlobe 1881:1889 arg1 communication 1891:1903 the interlobe communication 1877:1903 the interlobe communication 1877:1903 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 5 60 theme N-linked 879:886 arg1 chains 903:908 two N-linked complex glycan chains 875:908 two N-linked complex glycan chains located in the C-lobe 875:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 11 61 from separation 1690:1699 arg1 C-lobe 1704:1709 C-lobe 1704:1709 C-lobe 1704:1709 Results revealed that separation in C-lobe caused a significant structural variation in comparison to N-lobe. 27235585 3 62 theme dynamic 579:585 arg1 simulation 587:596 molecular dynamic simulation 569:596 molecular dynamic simulation 569:596 In the present work, molecular dynamic simulation was employed to gain access into the molecular level understanding of carbonate binding site and their interactions in each lobe. 27235585 5 63 theme glycan 896:901 arg1 chains 903:908 two N-linked complex glycan chains 875:908 two N-linked complex glycan chains located in the C-lobe 875:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 12 64 from importance 1863:1872 arg1 other 1941:1945 other 1941:1945 other 1941:1945 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 0 65 theme dynamic 70:76 arg1 analysis 89:96 A molecular dynamic simulation analysis 58:96 A molecular dynamic simulation analysis 58:96 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 1 66 theme cooperative 144:154 arg1 binding 156:162 the highly cooperative binding 133:162 the highly cooperative binding of iron 133:170 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 0 67 from effect 4:9 arg1 structure 47:55 the transferrin structure 31:55 the transferrin structure 31:55 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 11 68 theme structural 1732:1741 arg1 variation 1743:1751 a significant structural variation 1718:1751 a significant structural variation 1718:1751 Results revealed that separation in C-lobe caused a significant structural variation in comparison to N-lobe. 27235585 6 69 theme molecular 949:957 arg1 simulation 967:976 the molecular dynamic simulation 945:976 the molecular dynamic simulation for simplifying 945:992 Usually, in the molecular dynamic simulation for simplifying, glycan is removed from the protein structure. 27235585 5 70 dep chains 903:908 arg1 transferrin 840:850 native human transferrin 827:850 native human transferrin 827:850 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 5 70 dep chains 903:908 arg1 glycoprotein 857:868 a glycoprotein 855:868 a glycoprotein that two N-linked complex glycan chains located in the C-lobe 855:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 9 71 theme entire 1258:1263 arg1 transferrin 1265:1275 sometimes the entire transferrin 1244:1275 sometimes the entire transferrin 1244:1275 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 2 72 with interaction 463:473 arg1 transferrin 493:503 transferrin 493:503 transferrin 493:503 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 0 73 dep analysis 89:96 arg1 effect 4:9 The effect 0:9 The effect of glycosylation on the transferrin structure 0:55 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 2 74 theme carbonate 478:486 arg1 interaction 463:473 the interaction 459:473 the interaction of carbonate with transferrin 459:503 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 4 75 theme responsible 737:747 arg1 Residues 728:735 Residues 728:735 Residues responsible for carbonate binding of transferrin structure 728:794 Residues responsible for carbonate binding of transferrin structure were pointed out. 27235585 2 76 theme binding 354:360 arg1 site 362:365 C-lobe binding site 347:365 C-lobe binding site of the transferrin structure 347:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 2 76 theme binding 354:360 arg1 structure 386:394 the transferrin structure 370:394 the transferrin structure 370:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 5 77 theme located 910:916 arg1 chains 903:908 two N-linked complex glycan chains 875:908 two N-linked complex glycan chains located in the C-lobe 875:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 0 78 theme glycosylation 14:26 arg1 effect 4:9 The effect 0:9 The effect of glycosylation on the transferrin structure 0:55 The effect of glycosylation on the transferrin structure: A molecular dynamic simulation analysis. 27235585 9 79 theme separated 1287:1295 arg1 lobes 1297:1301 separated lobes 1287:1301 separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein 1287:1435 On the other hand, sometimes the entire transferrin formed by separated lobes that it allows the results to be interpreted in a straightforward manner rather than more parameters required for full length protein. 27235585 12 80 theme full 1820:1823 arg1 length 1825:1830 the full length one 1816:1834 the full length one 1816:1834 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 12 80 theme full 1820:1823 arg1 different 1840:1848 different 1840:1848 different 1840:1848 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 5 81 from glycoprotein 857:868 arg1 addition 817:824 addition 817:824 addition 817:824 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 2 82 theme transferrin 374:384 arg1 structure 386:394 the transferrin structure 370:394 the transferrin structure 370:394 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 7 83 from effect 1062:1067 arg1 structure 1105:1113 the transferrin structure 1089:1113 the transferrin structure 1089:1113 Here, we explore the effect of glycosylation on the transferrin structure. 27235585 4 84 theme structure 786:794 arg1 binding 763:769 carbonate binding 753:769 carbonate binding of transferrin structure 753:794 Residues responsible for carbonate binding of transferrin structure were pointed out. 27235585 10 85 theme dynamic 1592:1598 arg1 simulation 1600:1609 the molecular dynamic simulation 1578:1609 the molecular dynamic simulation 1578:1609 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 12 86 theme separated 1796:1804 arg1 lobes 1806:1810 the separated lobes 1792:1810 the separated lobes 1792:1810 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 12 86 theme separated 1796:1804 arg1 different 1840:1848 different 1840:1848 different 1840:1848 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 12 87 from impact 1913:1918 arg1 other 1941:1945 other 1941:1945 other 1941:1945 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 27235585 10 88 theme such 1640:1643 arg1 variations 1656:1665 such structural variations 1640:1665 such structural variations 1640:1665 But, it should be noted that there are differences between the separated lobe and full length transferrin, hence, a comparative analysis by the molecular dynamic simulation was performed to investigate such structural variations. 27235585 5 89 link N-linked 879:886 arg1 chains 903:908 two N-linked complex glycan chains 875:908 two N-linked complex glycan chains located in the C-lobe 875:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 4 90 theme carbonate 753:761 arg1 binding 763:769 carbonate binding 753:769 carbonate binding of transferrin structure 753:794 Residues responsible for carbonate binding of transferrin structure were pointed out. 27235585 2 91 theme transferrin 288:298 arg1 function 300:307 transferrin function 288:307 transferrin function 288:307 As such, the layout of the binding site residues affects transferrin function significantly; In contrast to N-lobe, C-lobe binding site of the transferrin structure has been less characterized and little research which surveyed the interaction of carbonate with transferrin in the C-lobe binding site has been found. 27235585 5 92 from located 910:916 arg1 C-lobe 925:930 the C-lobe 921:930 the C-lobe 921:930 In addition, native human transferrin is a glycoprotein that two N-linked complex glycan chains located in the C-lobe. 27235585 1 93 theme Fe-CO3-Tf 204:212 arg1 complex 222:228 a Fe-CO3-Tf ternary complex 202:228 a Fe-CO3-Tf ternary complex 202:228 Transferrins have been defined by the highly cooperative binding of iron and a carbonate anion to form a Fe-CO3-Tf ternary complex. 27235585 12 94 theme transferrin 1954:1964 arg1 structure 1966:1974 the transferrin structure 1950:1974 the transferrin structure 1950:1974 Consequently, the separated lobes and the full length one are different, showing the importance of the interlobe communication and the impact of the lobes on each other in the transferrin structure. 28661444 2 0 theme glycosylation 586:598 arg1 effect 563:568 the effect 559:568 the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 559:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 1 1 theme antigenic 400:408 arg1 proteins 410:417 antigenic proteins 400:417 antigenic proteins from Mycobacterium tuberculosis (MTB) 400:455 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 4 2 theme anomeric 878:885 arg1 position 887:894 the anomeric position 874:894 the anomeric position with a thiocyanomethyl group 874:923 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 2 3 from effect 563:568 arg1 proteins 623:630 two recombinant MTB proteins 603:630 two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 603:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 10 4 theme T-cell 1758:1763 arg1 activity 1765:1772 the T-cell activity 1754:1772 the T-cell activity 1754:1772 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 12 5 theme antibody 1983:1990 arg1 interaction 1992:2002 the antibody interaction 1979:2002 the antibody interaction of Ag85B 1979:2011 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 2 6 theme MTB 619:621 arg1 proteins 623:630 two recombinant MTB proteins 603:630 two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 603:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 5 7 dep ex 1032:1033 arg1 vivo 1035:1038 vivo 1035:1038 vivo 1035:1038 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 7 8 from reduction 1308:1316 arg1 response 1332:1339 the T-cell response 1321:1339 the T-cell response 1321:1339 Similarly, Ag85B maintains its B-cell activity after glycosylation while showing a significant reduction in the T-cell response. 28661444 3 9 theme Different 724:732 arg1 derivatives 765:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives were prepared, starting from mannose and two disaccharide analogs. 28661444 6 10 theme immunological 1167:1179 arg1 activity 1181:1188 the immunological activity 1163:1188 the immunological activity of the TB10.4 protein 1163:1210 Glycosylation does not modify the immunological activity of the TB10.4 protein. 28661444 5 11 gly neo-glycoproteins 1096:1112 arg1 neo-glycoproteins 1096:1112 the different neo-glycoproteins 1082:1112 the different neo-glycoproteins 1082:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 10 12 theme glycosylation 1587:1599 arg1 site 1601:1604 the main glycosylation site 1578:1604 the main glycosylation site 1578:1604 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 0 13 theme Ex 126:127 arg1 Evaluation 145:154 Ex Vivo Biological Evaluation 126:154 Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates 126:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 5 14 theme activity 1070:1077 arg1 evaluation 1040:1049 the ex vivo evaluation 1028:1049 the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins 1028:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 14 theme activity 1070:1077 arg1 sites 1018:1022 The glycosylation sites 1000:1022 The glycosylation sites 1000:1022 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 14 theme activity 1070:1077 arg1 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 2 15 gly glycosylation 586:598 arg1 MTB 619:621 two recombinant MTB proteins 603:630 two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 603:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 11 16 gly glycosylated 1871:1882 arg1 lysines 1799:1805 additional lysines 1788:1805 additional lysines included in different epitopes (Lys103, -123 and -282) 1788:1860 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 3 17 theme glycoconjugated 749:763 arg1 derivatives 765:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives were prepared, starting from mannose and two disaccharide analogs. 28661444 4 18 theme thiocyanomethyl 903:917 arg1 group 919:923 a thiocyanomethyl group 901:923 a thiocyanomethyl group 901:923 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 2 19 theme seven-amino 671:681 arg1 tag 688:690 an additional seven-amino acid tag 657:690 an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 657:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 0 20 theme Semi-Synthetic 163:176 arg1 Glycoconjugates 178:192 New Semi-Synthetic Glycoconjugates 159:192 New Semi-Synthetic Glycoconjugates 159:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 1 21 theme Mycobacterium 424:436 arg1 MTB 452:454 MTB 452:454 MTB 452:454 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 21 theme Mycobacterium 424:436 arg1 tuberculosis 438:449 Mycobacterium tuberculosis 424:449 Mycobacterium tuberculosis (MTB) 424:455 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 8 22 dep in 1445:1446 arg1 silico 1448:1453 silico 1448:1453 silico 1448:1453 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 1 23 from tuberculosis 438:449 arg1 proteins 410:417 antigenic proteins 400:417 antigenic proteins from Mycobacterium tuberculosis (MTB) 400:455 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 9 24 theme T-cell 1532:1537 arg1 epitope 1539:1545 any T-cell epitope 1528:1545 any T-cell epitope 1528:1545 In the recombinant TB10.4, the unique lysine is not included in any T-cell epitope. 28661444 2 25 theme recombinant 693:703 arg1 Ag85B 705:709 recombinant Ag85B 693:709 recombinant Ag85B 693:709 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 3 26 theme disaccharide 822:833 arg1 analogs 835:841 two disaccharide analogs 818:841 two disaccharide analogs 818:841 Different semi-synthetic glycoconjugated derivatives were prepared, starting from mannose and two disaccharide analogs. 28661444 7 27 theme B-cell 1244:1249 arg1 activity 1251:1258 its B-cell activity 1240:1258 its B-cell activity 1240:1258 Similarly, Ag85B maintains its B-cell activity after glycosylation while showing a significant reduction in the T-cell response. 28661444 12 28 theme epitopic 1905:1912 arg1 lysines 1914:1920 B-cell epitopic lysines 1898:1920 B-cell epitopic lysines of Ag85B 1898:1929 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 0 29 dep Ex 126:127 arg1 Biological 134:143 Biological 134:143 Biological 134:143 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 29 dep Ex 126:127 arg1 Vivo 129:132 Vivo 129:132 Vivo 129:132 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 4 30 theme protein 942:948 arg1 glycosylation 950:962 protein glycosylation 942:962 protein glycosylation 942:962 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 0 31 from Glycosylation 0:12 arg1 tuberculosis 67:78 Mycobacterium tuberculosis 53:78 Mycobacterium tuberculosis 53:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 8 32 theme putative 1379:1386 arg1 B- 1388:1389 putative B- 1379:1389 putative B- 1379:1389 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 5 33 theme neo-glycoproteins 1096:1112 arg1 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 0 34 theme Epitopes 113:120 arg1 Prediction 91:100 In Silico Prediction 81:100 In Silico Prediction of Protein Epitopes 81:120 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 34 theme Epitopes 113:120 arg1 Evaluation 145:154 Ex Vivo Biological Evaluation 126:154 Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates 126:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 10 35 theme T-cell 1673:1678 arg1 epitopes 1680:1687 T-cell epitopes 1673:1687 T-cell epitopes 1673:1687 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 0 36 theme Recombinant 17:27 arg1 Proteins 39:46 Recombinant Antigenic Proteins 17:46 Recombinant Antigenic Proteins from Mycobacterium tuberculosis 17:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 1 37 theme antigens 297:304 arg1 strategy 485:492 a new strategy 479:492 a new strategy for developing efficient vaccines 479:526 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 37 theme antigens 297:304 arg1 use 279:281 the use 275:281 the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), 275:456 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 2 38 dep tag 688:690 arg1 Ag85B 705:709 recombinant Ag85B 693:709 recombinant Ag85B 693:709 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 2 38 dep tag 688:690 arg1 TB10.4 715:720 TB10.4 715:720 TB10.4 715:720 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 0 39 theme Proteins 39:46 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 0:193 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 1 40 theme new 481:483 arg1 strategy 485:492 a new strategy 479:492 a new strategy for developing efficient vaccines 479:526 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 40 theme new 481:483 arg1 use 279:281 the use 275:281 the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), 275:456 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 0 41 theme Mycobacterium 53:65 arg1 tuberculosis 67:78 Mycobacterium tuberculosis 53:78 Mycobacterium tuberculosis 53:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 4 42 with reaction 977:984 arg1 lysines 991:997 lysines 991:997 lysines 991:997 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 1 43 theme efficient 509:517 arg1 vaccines 519:526 efficient vaccines 509:526 efficient vaccines 509:526 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 44 theme diseases 251:258 arg1 worldwide 260:268 the most deadly infectious diseases worldwide 224:268 the most deadly infectious diseases worldwide 224:268 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 0 45 dep Glycosylation 0:12 arg1 Evaluation 145:154 Ex Vivo Biological Evaluation 126:154 Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates 126:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 45 dep Glycosylation 0:12 arg1 Prediction 91:100 In Silico Prediction 81:100 In Silico Prediction of Protein Epitopes 81:120 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 12 46 theme Ag85B 2007:2011 arg1 interaction 1992:2002 the antibody interaction 1979:2002 the antibody interaction of Ag85B 1979:2011 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 2 47 theme chemical 577:584 arg1 glycosylation 586:598 the chemical glycosylation 573:598 the chemical glycosylation 573:598 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 10 48 theme main 1582:1585 arg1 site 1601:1604 the main glycosylation site 1578:1604 the main glycosylation site 1578:1604 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 12 49 gly glycosylated 1955:1966 arg1 lysines 1914:1920 B-cell epitopic lysines 1898:1920 B-cell epitopic lysines of Ag85B 1898:1929 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 6 50 theme protein 1204:1210 arg1 activity 1181:1188 the immunological activity 1163:1188 the immunological activity of the TB10.4 protein 1163:1210 Glycosylation does not modify the immunological activity of the TB10.4 protein. 28661444 12 51 with coupling 2048:2055 arg1 disaccharides 2071:2083 disaccharides 2071:2083 disaccharides 2071:2083 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 12 51 with coupling 2048:2055 arg1 mono- 2062:2066 mono- 2062:2066 mono- 2062:2066 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 2 52 theme recombinant 607:617 arg1 proteins 623:630 two recombinant MTB proteins 603:630 two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 603:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 9 53 theme recombinant 1471:1481 arg1 TB10.4 1483:1488 the recombinant TB10.4 1467:1488 the recombinant TB10.4 1467:1488 In the recombinant TB10.4, the unique lysine is not included in any T-cell epitope. 28661444 3 54 theme semi-synthetic 734:747 arg1 derivatives 765:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives 724:775 Different semi-synthetic glycoconjugated derivatives were prepared, starting from mannose and two disaccharide analogs. 28661444 9 55 theme unique 1495:1500 arg1 lysine 1502:1507 the unique lysine 1491:1507 the unique lysine 1491:1507 In the recombinant TB10.4, the unique lysine is not included in any T-cell epitope. 28661444 0 56 theme New 159:161 arg1 Glycoconjugates 178:192 New Semi-Synthetic Glycoconjugates 159:192 New Semi-Synthetic Glycoconjugates 159:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 5 57 theme different 1086:1094 arg1 neo-glycoproteins 1096:1112 the different neo-glycoproteins 1082:1112 the different neo-glycoproteins 1082:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 2 58 theme acid 683:686 arg1 tag 688:690 an additional seven-amino acid tag 657:690 an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 657:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 0 59 theme Glycoconjugates 178:192 arg1 Prediction 91:100 In Silico Prediction 81:100 In Silico Prediction of Protein Epitopes 81:120 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 59 theme Glycoconjugates 178:192 arg1 Evaluation 145:154 Ex Vivo Biological Evaluation 126:154 Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates 126:192 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 11 60 theme different 1819:1827 arg1 epitopes 1829:1836 different epitopes 1819:1836 different epitopes (Lys103, -123 and -282) 1819:1860 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 0 61 theme In 81:82 arg1 Prediction 91:100 In Silico Prediction 81:100 In Silico Prediction of Protein Epitopes 81:120 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 5 62 theme immunogenic 1058:1068 arg1 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 4 63 theme selective 967:975 arg1 reaction 977:984 selective reaction 967:984 selective reaction with lysines 967:997 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 11 64 dep epitopes 1829:1836 arg1 -282 1856:1859 -282 1856:1859 -282 1856:1859 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 11 64 dep epitopes 1829:1836 arg1 -123 1847:1850 -123 1847:1850 -123 1847:1850 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 11 64 dep epitopes 1829:1836 arg1 Lys103 1839:1844 Lys103 1839:1844 Lys103 1839:1844 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 11 65 theme additional 1788:1797 arg1 lysines 1799:1805 additional lysines 1788:1805 additional lysines included in different epitopes (Lys103, -123 and -282) 1788:1860 Furthermore, additional lysines included in different epitopes (Lys103, -123 and -282) are also glycosylated. 28661444 0 66 theme Protein 105:111 arg1 Epitopes 113:120 Protein Epitopes 105:120 Protein Epitopes 105:120 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 12 67 located found 1936:1940 arg2 lysines 1914:1920 B-cell epitopic lysines 1898:1920 B-cell epitopic lysines of Ag85B 1898:1929 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 12 67 located found 1936:1940 arg1 contrast 1888:1895 contrast 1888:1895 contrast 1888:1895 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 12 68 theme Ag85B 1925:1929 arg1 lysines 1914:1920 B-cell epitopic lysines 1898:1920 B-cell epitopic lysines of Ag85B 1898:1929 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 0 69 from tuberculosis 67:78 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 0:193 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 69 from tuberculosis 67:78 arg1 Proteins 39:46 Recombinant Antigenic Proteins 17:46 Recombinant Antigenic Proteins from Mycobacterium tuberculosis 17:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 2 70 theme additional 660:669 arg1 tag 688:690 an additional seven-amino acid tag 657:690 an additional seven-amino acid tag (recombinant Ag85B and TB10.4) 657:721 In this work, we investigated the effect of the chemical glycosylation on two recombinant MTB proteins produced in E. coli with an additional seven-amino acid tag (recombinant Ag85B and TB10.4). 28661444 8 71 theme B- 1388:1389 arg1 epitopes 1402:1409 the putative B- and T-cell epitopes 1375:1409 the putative B- and T-cell epitopes 1375:1409 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 12 72 theme B-cell 1898:1903 arg1 lysines 1914:1920 B-cell epitopic lysines 1898:1920 B-cell epitopic lysines of Ag85B 1898:1929 In contrast, B-cell epitopic lysines of Ag85B were found to be poorly glycosylated and, thus, the antibody interaction of Ag85B was only marginally affected after coupling with mono- or disaccharides. 28661444 10 73 gly glycosylation 1587:1599 arg2 site 1601:1604 the main glycosylation site 1578:1604 the main glycosylation site 1578:1604 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 8 74 theme T-cell 1395:1400 arg1 epitopes 1402:1409 the putative B- and T-cell epitopes 1375:1409 the putative B- and T-cell epitopes 1375:1409 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 10 75 theme important 1629:1637 arg1 Lys30 1548:1552 Lys30 1548:1552 Lys30 1548:1552 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 10 75 theme important 1629:1637 arg1 site 1639:1642 the most important site 1620:1642 the most important site involved in the formation of T-cell epitopes 1620:1687 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 0 76 theme Antigenic 29:37 arg1 Proteins 39:46 Recombinant Antigenic Proteins 17:46 Recombinant Antigenic Proteins from Mycobacterium tuberculosis 17:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 1 77 theme conjugated 286:295 arg1 antigens 297:304 conjugated antigens 286:304 conjugated antigens 286:304 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 0 78 dep In 81:82 arg1 Silico 84:89 Silico 84:89 Silico 84:89 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 0 79 gly Glycosylation 0:12 arg1 Proteins 39:46 Recombinant Antigenic Proteins 17:46 Recombinant Antigenic Proteins from Mycobacterium tuberculosis 17:78 Glycosylation of Recombinant Antigenic Proteins from Mycobacterium tuberculosis: In Silico Prediction of Protein Epitopes and Ex Vivo Biological Evaluation of New Semi-Synthetic Glycoconjugates. 28661444 4 80 with position 887:894 arg1 group 919:923 a thiocyanomethyl group 901:923 a thiocyanomethyl group 901:923 The glycans were activated at the anomeric position with a thiocyanomethyl group, as required for protein glycosylation by selective reaction with lysines. 28661444 7 81 theme T-cell 1325:1330 arg1 response 1332:1339 the T-cell response 1321:1339 the T-cell response 1321:1339 Similarly, Ag85B maintains its B-cell activity after glycosylation while showing a significant reduction in the T-cell response. 28661444 5 82 theme ex 1032:1033 arg1 evaluation 1040:1049 the ex vivo evaluation 1028:1049 the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins 1028:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 83 gly glycosylation 1004:1016 arg2 sites 1018:1022 The glycosylation sites 1000:1022 The glycosylation sites 1000:1022 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 83 gly glycosylation 1004:1016 arg2 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 83 gly glycosylation 1004:1016 arg1 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 1 84 theme antigenic 329:337 arg1 lipoarabinomannane 369:386 the lipoarabinomannane 365:386 the lipoarabinomannane (LAM) 365:392 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 84 theme antigenic 329:337 arg1 oligosaccharides 339:354 antigenic oligosaccharides 329:354 antigenic oligosaccharides 329:354 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 7 85 theme significant 1296:1306 arg1 reduction 1308:1316 a significant reduction 1294:1316 a significant reduction in the T-cell response 1294:1339 Similarly, Ag85B maintains its B-cell activity after glycosylation while showing a significant reduction in the T-cell response. 28661444 8 86 theme in 1445:1446 arg1 systems 1455:1461 in silico systems 1445:1461 in silico systems 1445:1461 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 10 87 theme epitopes 1680:1687 arg1 formation 1660:1668 the formation 1656:1668 the formation of T-cell epitopes 1656:1687 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 1 88 theme infectious 240:249 arg1 diseases 251:258 the most deadly infectious diseases 224:258 the most deadly infectious diseases worldwide 224:268 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 5 89 theme glycosylation 1004:1016 arg1 sites 1018:1022 The glycosylation sites 1000:1022 The glycosylation sites 1000:1022 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 5 89 theme glycosylation 1004:1016 arg1 activity 1070:1077 the immunogenic activity 1054:1077 the immunogenic activity of the different neo-glycoproteins 1054:1112 The glycosylation sites and the ex vivo evaluation of the immunogenic activity of the different neo-glycoproteins were investigated. 28661444 10 90 theme Ag85B 1557:1561 arg1 Lys30 1548:1552 Lys30 1548:1552 Lys30 1548:1552 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 10 90 theme Ag85B 1557:1561 arg1 site 1639:1642 the most important site 1620:1642 the most important site involved in the formation of T-cell epitopes 1620:1687 Lys30 of Ag85B, identified as the main glycosylation site, proved to be the most important site involved in the formation of T-cell epitopes, reasonably explaining why its glycosylation strongly influenced the T-cell activity. 28661444 8 91 theme systems 1455:1461 arg1 combination 1430:1440 a combination 1428:1440 a combination of in silico systems 1428:1461 The results were correlated with the putative B- and T-cell epitopes, predicted using a combination of in silico systems. 28661444 1 92 theme worldwide 260:268 arg1 worldwide 260:268 the most deadly infectious diseases worldwide 224:268 the most deadly infectious diseases worldwide 224:268 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28661444 1 92 theme worldwide 260:268 arg1 one 217:219 one 217:219 one 217:219 Tuberculosis is still one of the most deadly infectious diseases worldwide, and the use of conjugated antigens, obtained by combining antigenic oligosaccharides, such as the lipoarabinomannane (LAM), with antigenic proteins from Mycobacterium tuberculosis (MTB), has been proposed as a new strategy for developing efficient vaccines. 28122822 0 0 theme differential 81:92 arg1 expression 94:103 differential expression 81:103 differential expression of glycosyltranferases 81:126 Intestinal Muc2 mucin O-glycosylation is affected by microbiota and regulated by differential expression of glycosyltranferases. 28122822 9 1 theme O-glycosyltransferase 1259:1279 arg1 abundances 1281:1290 The O-glycosyltransferase abundances 1255:1290 The O-glycosyltransferase abundances along the intestine 1255:1310 The O-glycosyltransferase abundances along the intestine correlated well with the Muc2 O-glycan patterns. 28122822 4 2 theme inner 521:525 arg1 penetrable 542:551 penetrable 542:551 penetrable 542:551 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 4 2 theme inner 521:525 arg1 mucus 533:537 the inner colon mucus 517:537 the inner colon mucus 517:537 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 2 3 from intestine 181:189 arg1 present 221:227 present 221:227 present 221:227 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 3 from intestine 181:189 arg1 mucus 212:216 a single unattached mucus 192:216 a single unattached mucus 192:216 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 4 4 theme small 460:464 arg1 mucus 477:481 small intestinal mucus 460:481 small intestinal mucus 460:481 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 6 5 theme main 792:795 arg1 component 797:805 its main component 788:805 its main component the Muc2 mucin 788:820 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 2 6 contain has 247:249 arg2 mucus 305:309 an outer loose mucus 290:309 an outer loose mucus 290:309 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 6 contain has 247:249 arg2 layer 280:284 an inner attached mucus layer 256:284 an inner attached mucus layer 256:284 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 6 contain has 247:249 arg1 colon 241:245 the colon 237:245 the colon 237:245 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 8 7 theme simplified 1121:1130 arg1 assumption 1132:1141 the simplified assumption 1117:1141 the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis 1117:1252 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 10 8 theme GF 1448:1449 arg1 something 1457:1465 something 1457:1465 something that is in concordance with the observed shorter Muc2 O-glycans 1457:1529 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 10 8 theme GF 1448:1449 arg1 mice 1451:1454 GF mice 1448:1454 GF mice 1448:1454 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 5 9 theme host-microbiota 601:615 arg1 interactions 617:628 the host-microbiota interactions 597:628 the host-microbiota interactions 597:628 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 3 10 theme colon 338:342 arg1 impenetrable 347:358 impenetrable 347:358 impenetrable to bacteria 347:370 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 3 10 theme colon 338:342 arg1 mucus 325:329 The attached mucus 312:329 The attached mucus of the colon 312:342 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 4 11 theme germ-free 439:447 arg1 mice 454:457 germ-free (GF) mice 439:457 germ-free (GF) mice 439:457 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 2 12 theme mucus 274:278 arg1 layer 280:284 an inner attached mucus layer 256:284 an inner attached mucus layer 256:284 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 13 from present 221:227 arg1 intestine 181:189 the small intestine 171:189 the small intestine 171:189 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 14 located present 221:227 arg1 intestine 181:189 the small intestine 171:189 the small intestine 171:189 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 14 located present 221:227 arg2 mucus 212:216 a single unattached mucus 192:216 a single unattached mucus 192:216 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 14 located present 221:227 arg2 present 221:227 present 221:227 present 221:227 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 5 15 theme attachment 692:701 arg1 glycans 660:666 glycans 660:666 glycans 660:666 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 5 15 theme attachment 692:701 arg1 sites 703:707 attachment sites 692:707 attachment sites 692:707 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 7 16 theme epithelial 937:946 arg1 cells 948:952 the epithelial cells 933:952 the epithelial cells 933:952 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 2 17 theme attached 265:272 arg1 layer 280:284 an inner attached mucus layer 256:284 an inner attached mucus layer 256:284 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 0 18 theme glycosyltranferases 108:126 arg1 expression 94:103 differential expression 81:103 differential expression of glycosyltranferases 81:126 Intestinal Muc2 mucin O-glycosylation is affected by microbiota and regulated by differential expression of glycosyltranferases. 28122822 4 19 theme GF 450:451 arg1 mice 454:457 germ-free (GF) mice 439:457 germ-free (GF) mice 439:457 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 2 20 theme inner 259:263 arg1 layer 280:284 an inner attached mucus layer 256:284 an inner attached mucus layer 256:284 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 7 21 from intestine 964:972 arg1 mice 1006:1009 GF and conventionally raised mice 977:1009 mice 1006:1009 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 0 22 theme Muc2 11:14 arg1 O-glycosylation 22:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation is affected by microbiota and regulated by differential expression of glycosyltranferases. 28122822 6 23 theme O-glycan 842:849 arg1 patterns 851:858 regiospecific O-glycan patterns 828:858 regiospecific O-glycan patterns 828:858 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 5 24 theme important 579:587 arg1 role 589:592 an important role 576:592 an important role 576:592 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 2 25 theme loose 299:303 arg1 mucus 305:309 an outer loose mucus 290:309 an outer loose mucus 290:309 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 10 26 theme Muc2 1516:1519 arg1 O-glycans 1521:1529 the observed shorter Muc2 O-glycans 1495:1529 the observed shorter Muc2 O-glycans 1495:1529 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 0 27 theme Intestinal 0:9 arg1 O-glycosylation 22:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation is affected by microbiota and regulated by differential expression of glycosyltranferases. 28122822 2 28 attach present 221:227 arg2 mucus 212:216 a single unattached mucus 192:216 a single unattached mucus 192:216 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 28 attach present 221:227 arg1 intestine 181:189 the small intestine 171:189 the small intestine 171:189 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 28 attach present 221:227 arg2 present 221:227 present 221:227 present 221:227 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 29 theme outer 293:297 arg1 mucus 305:309 an outer loose mucus 290:309 an outer loose mucus 290:309 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 8 30 dep assumption 1132:1141 arg1 involved 1215:1222 involved 1215:1222 are involved in Muc2 O-glycan biosynthesis 1211:1252 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 7 31 theme glycosyltransferase 886:904 arg1 concentrations 915:928 the glycosyltransferase relative concentrations 882:928 the glycosyltransferase relative concentrations in the epithelial cells 882:952 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 7 32 theme relative 906:913 arg1 concentrations 915:928 the glycosyltransferase relative concentrations 882:928 the glycosyltransferase relative concentrations in the epithelial cells 882:952 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 8 33 gly O-glycosylated 1076:1089 arg1 product 1091:1097 the main O-glycosylated product 1067:1097 the main O-glycosylated product in mucus 1067:1106 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 8 33 gly O-glycosylated 1076:1089 arg1 Muc2 1059:1062 Muc2 1059:1062 Muc2 1059:1062 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 8 34 theme O-glycan 1232:1239 arg1 biosynthesis 1241:1252 Muc2 O-glycan biosynthesis 1227:1252 Muc2 O-glycan biosynthesis 1227:1252 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 0 35 theme mucin 16:20 arg1 O-glycosylation 22:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation 0:36 Intestinal Muc2 mucin O-glycosylation is affected by microbiota and regulated by differential expression of glycosyltranferases. 28122822 9 36 theme Muc2 1337:1340 arg1 patterns 1351:1358 the Muc2 O-glycan patterns 1333:1358 the Muc2 O-glycan patterns 1333:1358 The O-glycosyltransferase abundances along the intestine correlated well with the Muc2 O-glycan patterns. 28122822 6 37 theme regiospecific 828:840 arg1 patterns 851:858 regiospecific O-glycan patterns 828:858 regiospecific O-glycan patterns 828:858 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 4 38 attach attached 486:493 arg1 epithelium 502:511 the epithelium 498:511 the epithelium 498:511 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 4 38 attach attached 486:493 arg2 mucus 477:481 small intestinal mucus 460:481 small intestinal mucus 460:481 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 4 38 attach attached 486:493 arg3 mice 454:457 germ-free (GF) mice 439:457 germ-free (GF) mice 439:457 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 8 39 theme Muc2 1227:1230 arg1 biosynthesis 1241:1252 Muc2 O-glycan biosynthesis 1227:1252 Muc2 O-glycan biosynthesis 1227:1252 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 3 40 theme loose 382:386 arg1 habitat 404:410 a habitat 402:410 a habitat for commensal bacteria 402:433 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 3 40 theme loose 382:386 arg1 mucus 388:392 the loose mucus 378:392 the loose mucus 378:392 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 6 41 dep component 797:805 arg1 mucin 816:820 the Muc2 mucin 807:820 its main component the Muc2 mucin 788:820 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 5 42 theme nutrient 671:678 arg1 glycans 660:666 glycans 660:666 glycans 660:666 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 5 42 theme nutrient 671:678 arg1 sources 680:686 nutrient sources 671:686 nutrient sources 671:686 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 2 43 theme small 175:179 arg1 intestine 181:189 the small intestine 171:189 the small intestine 171:189 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 6 44 theme protein 722:728 arg1 composition 730:740 mucus protein composition 716:740 mucus protein composition 716:740 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 3 45 theme commensal 416:424 arg1 bacteria 426:433 commensal bacteria 416:433 commensal bacteria 416:433 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 8 46 theme epithelial 1194:1203 arg1 cells 1205:1209 the epithelial cells 1190:1209 the epithelial cells 1190:1209 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 4 47 theme intestinal 466:475 arg1 mucus 477:481 small intestinal mucus 460:481 small intestinal mucus 460:481 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 8 48 theme O-glycosylated 1076:1089 arg1 product 1091:1097 the main O-glycosylated product 1067:1097 the main O-glycosylated product in mucus 1067:1106 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 8 48 theme O-glycosylated 1076:1089 arg1 Muc2 1059:1062 Muc2 1059:1062 Muc2 1059:1062 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 6 49 theme Muc2 811:814 arg1 mucin 816:820 the Muc2 mucin 807:820 its main component the Muc2 mucin 788:820 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 6 50 theme mucus 716:720 arg1 composition 730:740 mucus protein composition 716:740 mucus protein composition 716:740 While mucus protein composition is relatively homogenous along the intestine, its main component the Muc2 mucin shows regiospecific O-glycan patterns. 28122822 1 51 theme Intestinal 129:138 arg1 cells 140:144 Intestinal cells 129:144 Intestinal cells 129:144 Intestinal cells are covered by mucus. 28122822 8 52 theme main 1071:1074 arg1 product 1091:1097 the main O-glycosylated product 1067:1097 the main O-glycosylated product in mucus 1067:1106 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 8 52 theme main 1071:1074 arg1 Muc2 1059:1062 Muc2 1059:1062 Muc2 1059:1062 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 10 53 theme O-glycan 1410:1417 arg1 elongation 1419:1428 the O-glycan elongation 1406:1428 the O-glycan elongation 1406:1428 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 10 54 with concordance 1478:1488 arg1 O-glycans 1521:1529 the observed shorter Muc2 O-glycans 1495:1529 the observed shorter Muc2 O-glycans 1495:1529 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 5 55 theme commensal 637:645 arg1 bacteria 647:654 the commensal bacteria 633:654 the commensal bacteria 633:654 O-glycosylation plays an important role in the host-microbiota interactions as the commensal bacteria use glycans as nutrient sources and attachment sites. 28122822 2 56 theme unattached 201:210 arg1 present 221:227 present 221:227 present 221:227 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 56 theme unattached 201:210 arg1 mucus 212:216 a single unattached mucus 192:216 a single unattached mucus 192:216 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 8 57 located found 1181:1185 arg1 cells 1205:1209 the epithelial cells 1190:1209 the epithelial cells 1190:1209 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 8 57 located found 1181:1185 arg2 glycosyltransferases 1160:1179 the glycosyltransferases 1156:1179 the glycosyltransferases found in the epithelial cells 1156:1209 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 28122822 7 58 from concentrations 915:928 arg1 cells 948:952 the epithelial cells 933:952 the epithelial cells 933:952 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 10 59 theme observed 1499:1506 arg1 O-glycans 1521:1529 the observed shorter Muc2 O-glycans 1495:1529 the observed shorter Muc2 O-glycans 1495:1529 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 2 60 theme single 194:199 arg1 present 221:227 present 221:227 present 221:227 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 2 60 theme single 194:199 arg1 mucus 212:216 a single unattached mucus 192:216 a single unattached mucus 192:216 In the small intestine, a single unattached mucus is present whereas the colon has both an inner attached mucus layer and an outer loose mucus. 28122822 7 61 theme raised 999:1004 arg1 mice 1006:1009 GF and conventionally raised mice 977:1009 mice 1006:1009 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 10 62 theme shorter 1508:1514 arg1 O-glycans 1521:1529 the observed shorter Muc2 O-glycans 1495:1529 the observed shorter Muc2 O-glycans 1495:1529 Some of the glycosyltransferases involved in the O-glycan elongation were decreased in GF mice, something that is in concordance with the observed shorter Muc2 O-glycans. 28122822 7 63 theme GF 977:978 arg1 mice 1006:1009 GF and conventionally raised mice 977:1009 mice 1006:1009 We have now analyzed the glycosyltransferase relative concentrations in the epithelial cells along the intestine in GF and conventionally raised mice and compared this with the O-glycans formed. 28122822 9 64 theme O-glycan 1342:1349 arg1 patterns 1351:1358 the Muc2 O-glycan patterns 1333:1358 the Muc2 O-glycan patterns 1333:1358 The O-glycosyltransferase abundances along the intestine correlated well with the Muc2 O-glycan patterns. 28122822 4 65 theme colon 527:531 arg1 penetrable 542:551 penetrable 542:551 penetrable 542:551 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 4 65 theme colon 527:531 arg1 mucus 533:537 the inner colon mucus 517:537 the inner colon mucus 517:537 In germ-free (GF) mice, small intestinal mucus is attached to the epithelium and the inner colon mucus is penetrable. 28122822 3 66 theme attached 316:323 arg1 impenetrable 347:358 impenetrable 347:358 impenetrable to bacteria 347:370 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 3 66 theme attached 316:323 arg1 mucus 325:329 The attached mucus 312:329 The attached mucus of the colon 312:342 The attached mucus of the colon is impenetrable to bacteria while the loose mucus acts as a habitat for commensal bacteria. 28122822 8 67 from product 1091:1097 arg1 mucus 1102:1106 mucus 1102:1106 mucus 1102:1106 As Muc2 is the main O-glycosylated product in mucus, we made the simplified assumption that most of the glycosyltransferases found in the epithelial cells are involved in Muc2 O-glycan biosynthesis. 29268168 2 0 gly afucosylated 383:394 arg1 IgG 396:398 the afucosylated IgG 379:398 the afucosylated IgG 379:398 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 6 1 theme reduced 998:1004 arg1 fucose 1006:1011 reduced fucose 998:1011 reduced fucose 998:1011 However, unlike hFcγRIIIa, mFcγRIV did not bind more avidly to IgG with increased galactose and reduced fucose. 29268168 7 2 theme human 1068:1072 arg1 family 1082:1087 the human FcγRIII family 1064:1087 the human FcγRIII family 1064:1087 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 3 3 theme afucosylated 468:479 arg1 IgG 481:483 afucosylated IgG 468:483 afucosylated IgG 468:483 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 1 4 theme IgG 233:235 arg1 IgG 233:235 IgG 233:235 IgG 233:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 1 4 theme IgG 233:235 arg1 Fc-domain 220:228 the Fc-domain 216:228 the Fc-domain of IgG 216:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 4 5 gly glycosylation 700:712 arg2 site 714:717 the N162 glycosylation site 691:717 the N162 glycosylation site 691:717 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 3 6 theme N-linked 604:611 arg1 glycan 613:618 an N-linked glycan 601:618 an N-linked glycan 601:618 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 1 7 link N-linked 180:187 arg1 glycan 189:194 the N-linked glycan 176:194 the N-linked glycan 176:194 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 2 8 theme binding 280:286 arg1 affinity 288:295 the binding affinity 276:295 the binding affinity of human IgG1 to human FcγRIIIa 276:327 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 9 theme direct 524:529 arg1 interactions 557:568 direct carbohydrate-carbohydrate interactions 524:568 direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb 524:668 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 5 10 from N162-glycan 777:787 arg1 mFcγRIV 792:798 mFcγRIV 792:798 mFcγRIV 792:798 The N162-glycan in mFcγRIV was also responsible for enhancing the binding to mouse IgG with reduced fucose similar to hFcγRIIIa. 29268168 2 11 dep fold 433:436 arg1 ∼40 429:431 ∼40 429:431 ∼40 429:431 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 12 from position 623:630 arg1 glycan 589:594 the IgG-Fc glycan 578:594 the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb 578:668 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 7 13 theme mouse 1109:1113 arg1 FcγRIV 1115:1120 its orthologous mouse FcγRIV 1093:1120 its orthologous mouse FcγRIV 1093:1120 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 2 14 theme afucosylated 383:394 arg1 IgG 396:398 the afucosylated IgG 379:398 the afucosylated IgG 379:398 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 15 gly afucosylated 468:479 arg1 IgG 481:483 afucosylated IgG 468:483 afucosylated IgG 468:483 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 4 16 theme mouse 752:756 arg1 mFcγRIV 764:770 mFcγRIV 764:770 mFcγRIV 764:770 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 4 16 theme mouse 752:756 arg1 FcγR 758:761 the orthologous mouse FcγR 736:761 the orthologous mouse FcγR 736:761 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 7 17 from FcγRIV 1115:1120 arg1 family 1082:1087 the human FcγRIII family 1064:1087 the human FcγRIII family 1064:1087 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 2 18 theme additional 349:358 arg1 galactosylation 360:374 additional galactosylation 349:374 additional galactosylation of the afucosylated IgG 349:398 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 0 19 theme FcγR- 10:14 arg1 glycan 16:21 Conserved FcγR- glycan 0:21 Conserved FcγR- glycan 0:21 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 3 20 from increase 443:450 arg1 affinity 455:462 affinity 455:462 affinity for afucosylated IgG 455:483 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 2 21 theme human 300:304 arg1 IgG1 306:309 human IgG1 300:309 human IgG1 300:309 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 0 22 theme Conserved 0:8 arg1 glycan 16:21 Conserved FcγR- glycan 0:21 Conserved FcγR- glycan 0:21 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 4 23 located found 727:731 arg1 mFcγRIV 764:770 mFcγRIV 764:770 mFcγRIV 764:770 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 4 23 located found 727:731 arg1 FcγR 758:761 the orthologous mouse FcγR 736:761 the orthologous mouse FcγR 736:761 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 4 23 located found 727:731 arg2 site 714:717 the N162 glycosylation site 691:717 the N162 glycosylation site 691:717 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 2 24 dep affinity 414:421 arg1 fold 433:436 fold 433:436 fold 433:436 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 25 theme carbohydrate-carbohydrate 531:555 arg1 interactions 557:568 direct carbohydrate-carbohydrate interactions 524:568 direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb 524:668 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 7 26 theme orthologous 1097:1107 arg1 FcγRIV 1115:1120 its orthologous mouse FcγRIV 1093:1120 its orthologous mouse FcγRIV 1093:1120 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 4 27 theme glycosylation 700:712 arg1 site 714:717 the N162 glycosylation site 691:717 the N162 glycosylation site 691:717 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 2 28 theme IgG 396:398 arg1 galactosylation 360:374 additional galactosylation 349:374 additional galactosylation of the afucosylated IgG 349:398 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 29 link N-linked 604:611 arg1 glycan 613:618 an N-linked glycan 601:618 an N-linked glycan 601:618 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 4 30 theme orthologous 740:750 arg1 mFcγRIV 764:770 mFcγRIV 764:770 mFcγRIV 764:770 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 4 30 theme orthologous 740:750 arg1 FcγR 758:761 the orthologous mouse FcγR 736:761 the orthologous mouse FcγR 736:761 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 1 31 theme binding 102:108 arg1 strength 110:117 The binding strength 98:117 The binding strength between IgG and FcγR 98:138 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 1 32 theme glycan 189:194 arg1 composition 161:171 the composition 157:171 the composition of the N-linked glycan at position N297 in the Fc-domain of IgG 157:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 0 33 theme fucosylated 45:55 arg1 IgG 74:76 fucosylated and afucosylated IgG 45:76 fucosylated and afucosylated IgG in humans and mice 45:95 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 2 34 dep ∼40 429:431 arg1 to 426:427 to 426:427 to 426:427 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 3 35 theme IgG-Fc 582:587 arg1 glycan 589:594 the IgG-Fc glycan 578:594 the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb 578:668 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 6 36 theme increased 974:982 arg1 galactose 984:992 increased galactose 974:992 increased galactose 974:992 However, unlike hFcγRIIIa, mFcγRIV did not bind more avidly to IgG with increased galactose and reduced fucose. 29268168 0 37 theme afucosylated 61:72 arg1 IgG 74:76 fucosylated and afucosylated IgG 45:76 fucosylated and afucosylated IgG in humans and mice 45:95 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 7 38 from N162-glycan 1049:1059 arg1 family 1082:1087 the human FcγRIII family 1064:1087 the human FcγRIII family 1064:1087 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 0 39 from IgG 74:76 arg1 humans 81:86 humans 81:86 humans 81:86 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 0 39 from IgG 74:76 arg1 mice 92:95 mice 92:95 mice 92:95 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 1 40 from position 199:206 arg1 IgG 233:235 IgG 233:235 IgG 233:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 1 40 from position 199:206 arg1 Fc-domain 220:228 the Fc-domain 216:228 the Fc-domain of IgG 216:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 1 40 from position 199:206 arg1 composition 161:171 the composition 157:171 the composition of the N-linked glycan at position N297 in the Fc-domain of IgG 157:235 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 5 41 theme reduced 865:871 arg1 fucose 873:878 reduced fucose 865:878 reduced fucose similar to hFcγRIIIa 865:899 The N162-glycan in mFcγRIV was also responsible for enhancing the binding to mouse IgG with reduced fucose similar to hFcγRIIIa. 29268168 5 42 with binding 839:845 arg1 fucose 873:878 reduced fucose 865:878 reduced fucose similar to hFcγRIIIa 865:899 The N162-glycan in mFcγRIV was also responsible for enhancing the binding to mouse IgG with reduced fucose similar to hFcγRIIIa. 29268168 6 43 with IgG 965:967 arg1 galactose 984:992 increased galactose 974:992 increased galactose 974:992 However, unlike hFcγRIIIa, mFcγRIV did not bind more avidly to IgG with increased galactose and reduced fucose. 29268168 6 43 with IgG 965:967 arg1 fucose 1006:1011 reduced fucose 998:1011 reduced fucose 998:1011 However, unlike hFcγRIIIa, mFcγRIV did not bind more avidly to IgG with increased galactose and reduced fucose. 29268168 2 44 theme human 314:318 arg1 FcγRIIIa 320:327 human FcγRIIIa 314:327 human FcγRIIIa 314:327 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 4 45 theme N162 695:698 arg1 site 714:717 the N162 glycosylation site 691:717 the N162 glycosylation site 691:717 Here we report that the N162 glycosylation site is also found in the orthologous mouse FcγR, mFcγRIV. 29268168 3 46 theme unique 636:641 arg1 position 623:630 position 162 unique to hFcγRIIIa and hFcγRIIIb 623:668 position 162 unique to hFcγRIIIa and hFcγRIIIb 623:668 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 0 47 gly fucosylated 45:55 arg1 IgG 74:76 fucosylated and afucosylated IgG 45:76 fucosylated and afucosylated IgG in humans and mice 45:95 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 2 48 theme IgG1 306:309 arg1 affinity 288:295 the binding affinity 276:295 the binding affinity of human IgG1 to human FcγRIIIa 276:327 Particularly, afucosylation increases the binding affinity of human IgG1 to human FcγRIIIa up to ∼20 fold, and additional galactosylation of the afucosylated IgG increases the affinity up to ∼40 fold. 29268168 5 49 theme similar 880:886 arg1 fucose 873:878 reduced fucose 865:878 reduced fucose similar to hFcγRIIIa 865:899 The N162-glycan in mFcγRIV was also responsible for enhancing the binding to mouse IgG with reduced fucose similar to hFcγRIIIa. 29268168 3 50 with glycan 589:594 arg1 glycan 613:618 an N-linked glycan 601:618 an N-linked glycan 601:618 The increase in affinity for afucosylated IgG has previously been shown to depend on direct carbohydrate-carbohydrate interactions between the IgG-Fc glycan with an N-linked glycan at position 162 unique to hFcγRIIIa and hFcγRIIIb. 29268168 1 51 theme N-linked 180:187 arg1 glycan 189:194 the N-linked glycan 176:194 the N-linked glycan 176:194 The binding strength between IgG and FcγR is influenced by the composition of the N-linked glycan at position N297 in the Fc-domain of IgG. 29268168 7 52 theme FcγRIII 1074:1080 arg1 family 1082:1087 the human FcγRIII family 1064:1087 the human FcγRIII family 1064:1087 Overall, these results suggest the N162-glycan in the human FcγRIII family and its orthologous mouse FcγRIV to be functionally conserved. 29268168 0 53 gly afucosylated 61:72 arg1 IgG 74:76 fucosylated and afucosylated IgG 45:76 fucosylated and afucosylated IgG in humans and mice 45:95 Conserved FcγR- glycan discriminates between fucosylated and afucosylated IgG in humans and mice. 29268168 5 54 theme mouse 850:854 arg1 IgG 856:858 mouse IgG 850:858 mouse IgG 850:858 The N162-glycan in mFcγRIV was also responsible for enhancing the binding to mouse IgG with reduced fucose similar to hFcγRIIIa. 29055820 3 0 theme IgG 318:320 arg1 synthesis 329:337 IgG glycan synthesis 318:337 IgG glycan synthesis 318:337 IgG glycan synthesis is highly complex and dynamic. 29055820 10 1 theme DNA 1461:1463 arg1 methylation 1465:1475 DNA methylation 1461:1475 DNA methylation 1461:1475 Presumably, DNA methylation mediates the effect of smoking on IgG glycosylation. 29055820 6 2 dep overlap 639:645 arg1 cg06126421 912:921 cg06126421 912:921 cg06126421 912:921 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 cg03636183 938:947 cg03636183 938:947 cg03636183 938:947 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 chr.2 870:874 chr.2 870:874 chr.2 870:874 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 chr.19 950:955 chr.19 950:955 chr.19 950:955 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 chr.5 898:902 chr.5 898:902 chr.5 898:902 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 chr.1 816:820 chr.1 816:820 chr.1 816:820 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 6p21.33 924:930 6p21.33 924:930 6p21.33 924:930 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 cg05575921 886:895 cg05575921 886:895 cg05575921 886:895 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 2 dep overlap 639:645 arg1 cg01940273 858:867 cg01940273 858:867 cg01940273 858:867 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 8 3 theme DNA 1211:1213 arg1 methylation 1215:1225 DNA methylation 1211:1225 DNA methylation 1211:1225 Our results suggest the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures. 29055820 3 4 theme glycan 322:327 arg1 synthesis 329:337 IgG glycan synthesis 318:337 IgG glycan synthesis 318:337 IgG glycan synthesis is highly complex and dynamic. 29055820 2 5 theme effector 203:210 arg1 function 212:219 The effector function 199:219 The effector function of immunoglobulin G (IgG) 199:245 The effector function of immunoglobulin G (IgG) alters between pro- and anti-inflammatory, based on its glycosylation. 29055820 1 6 theme protein 167:173 arg1 structure 175:183 protein structure 167:183 protein structure 167:183 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 9 7 theme epigenome-wide 1302:1315 arg1 analysis 1317:1324 An epigenome-wide analysis 1299:1324 An epigenome-wide analysis conducted in four population-based cohorts 1299:1367 An epigenome-wide analysis conducted in four population-based cohorts revealed an association between DNA methylation and IgG glycosylation patterns. 29055820 10 8 theme smoking 1500:1506 arg1 effect 1490:1495 the effect 1486:1495 the effect of smoking on IgG glycosylation 1486:1527 Presumably, DNA methylation mediates the effect of smoking on IgG glycosylation. 29055820 6 9 theme IgG 785:787 arg1 glycosylation 789:801 IgG glycosylation 785:801 IgG glycosylation 785:801 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 4 10 theme analytical 400:409 arg1 methods 411:417 two different analytical methods 386:417 two different analytical methods for assessing IgG glycosylation 386:449 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 6 11 dep chr.2 870:874 arg1 ALPPL2 877:882 ALPPL2 877:882 ALPPL2 877:882 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 9 12 theme DNA 1401:1403 arg1 methylation 1405:1415 DNA methylation 1401:1415 DNA methylation 1401:1415 An epigenome-wide analysis conducted in four population-based cohorts revealed an association between DNA methylation and IgG glycosylation patterns. 29055820 1 13 from influences 153:162 arg1 function 189:196 function 189:196 function 189:196 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 1 13 from influences 153:162 arg1 structure 175:183 protein structure 167:183 protein structure 167:183 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 6 14 from panels 694:699 arg1 results 654:660 the results 650:660 the results from the two glycan measurement panels 650:699 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 15 theme measurement 682:692 arg1 panels 694:699 the two glycan measurement panels 667:699 the two glycan measurement panels 667:699 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 4 16 with aim 455:457 arg1 use 379:381 the use 375:381 the use of two different analytical methods for assessing IgG glycosylation 375:449 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 9 17 theme glycosylation 1425:1437 arg1 patterns 1439:1446 IgG glycosylation patterns 1421:1446 IgG glycosylation patterns 1421:1446 An epigenome-wide analysis conducted in four population-based cohorts revealed an association between DNA methylation and IgG glycosylation patterns. 29055820 6 18 dep chr.19 950:955 arg1 F2RL3 958:962 F2RL3 958:962 F2RL3 958:962 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 6 19 theme glycan 675:680 arg1 panels 694:699 the two glycan measurement panels 667:699 the two glycan measurement panels 667:699 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 5 20 from cohorts 612:618 arg1 individuals 593:603 3000 individuals 588:603 3000 individuals from 4 cohorts 588:618 In total, 3000 individuals from 4 cohorts were analyzed. 29055820 7 21 theme DNA 1105:1107 arg1 levels 1121:1126 DNA methylation levels 1105:1126 DNA methylation levels at these 7 CpG-sites 1105:1147 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 10 22 theme IgG 1511:1513 arg1 glycosylation 1515:1527 IgG glycosylation 1511:1527 IgG glycosylation 1511:1527 Presumably, DNA methylation mediates the effect of smoking on IgG glycosylation. 29055820 9 23 theme population-based 1344:1359 arg1 cohorts 1361:1367 four population-based cohorts 1339:1367 four population-based cohorts 1339:1367 An epigenome-wide analysis conducted in four population-based cohorts revealed an association between DNA methylation and IgG glycosylation patterns. 29055820 0 24 theme IgG 0:2 arg1 glycosylation 4:16 IgG glycosylation 0:16 IgG glycosylation 0:16 IgG glycosylation and DNA methylation are interconnected with smoking. 29055820 10 25 from effect 1490:1495 arg1 glycosylation 1515:1527 IgG glycosylation 1511:1527 IgG glycosylation 1511:1527 Presumably, DNA methylation mediates the effect of smoking on IgG glycosylation. 29055820 0 26 theme DNA 22:24 arg1 methylation 26:36 DNA methylation 22:36 DNA methylation 22:36 IgG glycosylation and DNA methylation are interconnected with smoking. 29055820 4 27 theme different 390:398 arg1 methods 411:417 two different analytical methods 386:417 two different analytical methods for assessing IgG glycosylation 386:449 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 4 28 theme association 557:567 arg1 studies 569:575 epigenome-wide association studies 542:575 epigenome-wide association studies 542:575 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 7 29 theme Mediation 966:974 arg1 analyses 976:983 Mediation analyses 966:983 Mediation analyses with respect to smoking 966:1007 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 6 30 from methylation 713:723 arg1 locations 753:761 5 genomic locations 743:761 5 genomic locations 743:761 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 8 31 theme link 1198:1201 arg1 presence 1174:1181 the presence 1170:1181 the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures 1170:1296 Our results suggest the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures. 29055820 7 32 theme smoking 1037:1043 arg1 effect 1027:1032 the effect 1023:1032 the effect of smoking on IgG glycosylation 1023:1064 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 4 33 theme epigenome-wide 542:555 arg1 studies 569:575 epigenome-wide association studies 542:575 epigenome-wide association studies 542:575 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 8 34 theme environmental 1274:1286 arg1 exposures 1288:1296 environmental exposures 1274:1296 environmental exposures 1274:1296 Our results suggest the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures. 29055820 7 35 theme IgG 1048:1050 arg1 glycosylation 1052:1064 IgG glycosylation 1048:1064 IgG glycosylation 1048:1064 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 2 36 theme G 239:239 arg1 function 212:219 The effector function 199:219 The effector function of immunoglobulin G (IgG) 199:245 The effector function of immunoglobulin G (IgG) alters between pro- and anti-inflammatory, based on its glycosylation. 29055820 6 37 theme genomic 745:751 arg1 locations 753:761 5 genomic locations 743:761 5 genomic locations 743:761 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 1 38 theme common 104:109 arg1 modifications 128:140 the most common post-translation modifications 95:140 the most common post-translation modifications 95:140 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 7 39 from effect 1027:1032 arg1 glycosylation 1052:1064 IgG glycosylation 1048:1064 IgG glycosylation 1048:1064 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 6 40 dep chr.5 898:902 arg1 AHRR 905:908 AHRR 905:908 AHRR 905:908 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 2 41 theme immunoglobulin 224:237 arg1 IgG 242:244 IgG 242:244 IgG 242:244 The effector function of immunoglobulin G (IgG) alters between pro- and anti-inflammatory, based on its glycosylation. 29055820 2 41 theme immunoglobulin 224:237 arg1 G 239:239 immunoglobulin G 224:239 immunoglobulin G (IgG) 224:245 The effector function of immunoglobulin G (IgG) alters between pro- and anti-inflammatory, based on its glycosylation. 29055820 6 42 theme CpG-sites 730:738 arg1 methylation 713:723 DNA methylation 709:723 DNA methylation of 7 CpG-sites on 5 genomic locations 709:761 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 1 43 theme post-translation 111:126 arg1 modifications 128:140 the most common post-translation modifications 95:140 the most common post-translation modifications 95:140 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 4 44 theme IgG 526:528 arg1 glycosylation 509:521 glycosylation 509:521 glycosylation of IgG by means of epigenome-wide association studies 509:575 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 4 44 theme IgG 526:528 arg1 methylation 493:503 DNA methylation 489:503 DNA methylation 489:503 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 4 45 theme methods 411:417 arg1 use 379:381 the use 375:381 the use of two different analytical methods for assessing IgG glycosylation 375:449 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 1 46 theme modifications 128:140 arg1 modifications 128:140 the most common post-translation modifications 95:140 the most common post-translation modifications 95:140 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 1 46 theme modifications 128:140 arg1 one 88:90 one 88:90 one 88:90 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 8 47 theme IgG 1231:1233 arg1 glycosylation 1235:1247 IgG glycosylation 1231:1247 IgG glycosylation 1231:1247 Our results suggest the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures. 29055820 4 48 theme IgG 433:435 arg1 glycosylation 437:449 IgG glycosylation 433:449 IgG glycosylation 433:449 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 4 49 gly glycosylation 509:521 arg1 IgG 526:528 IgG 526:528 IgG 526:528 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 9 50 theme IgG 1421:1423 arg1 patterns 1439:1446 IgG glycosylation patterns 1421:1446 IgG glycosylation patterns 1421:1446 An epigenome-wide analysis conducted in four population-based cohorts revealed an association between DNA methylation and IgG glycosylation patterns. 29055820 7 51 theme methylation 1109:1119 arg1 levels 1121:1126 DNA methylation levels 1105:1126 DNA methylation levels at these 7 CpG-sites 1105:1147 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 0 52 with interconnected 42:55 arg1 smoking 62:68 smoking 62:68 smoking 62:68 IgG glycosylation and DNA methylation are interconnected with smoking. 29055820 1 53 theme large 147:151 arg1 influences 153:162 large influences 147:162 large influences on protein structure and function 147:196 Glycosylation is one of the most common post-translation modifications with large influences on protein structure and function. 29055820 7 54 from CpG-sites 1139:1147 arg1 levels 1121:1126 DNA methylation levels 1105:1126 DNA methylation levels at these 7 CpG-sites 1105:1147 Mediation analyses with respect to smoking revealed that the effect of smoking on IgG glycosylation may be at least partially mediated via DNA methylation levels at these 7 CpG-sites. 29055820 4 55 theme DNA 489:491 arg1 methylation 493:503 DNA methylation 489:503 DNA methylation 489:503 With the use of two different analytical methods for assessing IgG glycosylation, we aim to elucidate the link between DNA methylation and glycosylation of IgG by means of epigenome-wide association studies. 29055820 6 56 theme DNA 709:711 arg1 methylation 713:723 DNA methylation 709:723 DNA methylation of 7 CpG-sites on 5 genomic locations 709:761 The overlap of the results from the two glycan measurement panels yielded DNA methylation of 7 CpG-sites on 5 genomic locations to be associated with IgG glycosylation: cg25189904 (chr.1, GNG12); cg05951221, cg21566642 and cg01940273 (chr.2, ALPPL2); cg05575921 (chr.5, AHRR); cg06126421 (6p21.33); and cg03636183 (chr.19, F2RL3). 29055820 8 57 theme indirect 1189:1196 arg1 link 1198:1201 an indirect link 1186:1201 an indirect link between DNA methylation and IgG glycosylation 1186:1247 Our results suggest the presence of an indirect link between DNA methylation and IgG glycosylation that may in part capture environmental exposures. 27808502 4 0 theme glycan 824:829 arg1 structure 831:839 The glycan structure 820:839 The glycan structure of intact glycopeptides 820:863 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 2 1 theme core 445:448 arg1 structures 458:467 core mannose structures 445:467 core mannose structures 445:467 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 1 2 with sugars 241:246 arg1 exoglycosidases 301:315 exoglycosidases 301:315 exoglycosidases 301:315 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 2 3 from MS/MS 536:540 arg1 spectrometry 550:561 mass spectrometry 545:561 mass spectrometry 545:561 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 5 4 from tolerance 1049:1057 arg1 time 1087:1090 retention time 1077:1090 retention time 1077:1090 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 4 5 theme MS3 966:968 arg1 spectra 970:976 the MS3 spectra 962:976 the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion 962:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 6 6 theme low 1197:1199 arg1 cost 1201:1204 low cost 1197:1204 low cost 1197:1204 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 4 7 theme oligomannosylated 985:1001 arg1 glycopeptides 1003:1015 the oligomannosylated glycopeptides 981:1015 the oligomannosylated glycopeptides with the same Y1 ion 981:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 6 8 theme cost 1201:1204 arg1 advantage 1184:1192 the advantage 1180:1192 the advantage of low cost and ease of processing 1180:1227 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 4 9 theme own 911:913 arg1 spectra 894:900 MS/MS spectra 888:900 MS/MS spectra of their own 888:913 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 7 10 theme N-glycopeptides 1509:1523 arg1 identification 1487:1500 the identification 1483:1500 the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1483:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 3 11 theme intact 647:652 arg1 N-glycopeptidome 654:669 the intact N-glycopeptidome 643:669 the intact N-glycopeptidome 643:669 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 5 12 theme retention 1077:1085 arg1 time 1087:1090 retention time 1077:1090 retention time 1077:1090 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 3 13 gly glycosylation 758:770 arg2 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 13 gly glycosylation 758:770 arg1 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 13 gly glycosylation 758:770 arg2 site 772:775 glycosylation site 758:775 glycosylation site 758:775 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 14 theme glycopeptides 805:817 arg1 site 772:775 glycosylation site 758:775 glycosylation site 758:775 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 14 theme glycopeptides 805:817 arg1 structure 785:793 the structure 781:793 the structure of intact glycopeptides 781:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 14 theme glycopeptides 805:817 arg1 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 14 theme glycopeptides 805:817 arg1 sequence 748:755 peptide sequence 740:755 peptide sequence 740:755 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 4 15 theme glycopeptides 1003:1015 arg1 spectra 970:976 the MS3 spectra 962:976 the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion 962:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 4 16 theme MS/MS 888:892 arg1 spectra 894:900 MS/MS spectra 888:900 MS/MS spectra of their own 888:913 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 4 17 theme intact 844:849 arg1 glycopeptides 851:863 intact glycopeptides 844:863 intact glycopeptides 844:863 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 1 18 gly N-glycopeptide 260:273 arg2 N-glycopeptide 260:273 the N-glycopeptide pentasaccharide core 256:294 the N-glycopeptide pentasaccharide core 256:294 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 2 19 theme Y1 510:511 arg1 ions 513:516 Y1 ions 510:516 Y1 ions 510:516 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 7 20 theme cells 1564:1568 arg1 lysates 1548:1554 the secretome and lysates 1530:1554 lysates 1548:1554 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 20 theme cells 1564:1568 arg1 secretome 1534:1542 the secretome and lysates 1530:1554 secretome 1534:1542 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 5 21 theme mass 1044:1047 arg1 tolerance 1049:1057 mass tolerance 1044:1057 mass tolerance 1044:1057 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 6 22 theme processing 1218:1227 arg1 ease 1210:1213 ease 1210:1213 ease of processing 1210:1227 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 6 22 theme processing 1218:1227 arg1 cost 1201:1204 low cost 1197:1204 low cost 1197:1204 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 4 23 gly oligomannosylated 985:1001 arg1 glycopeptides 1003:1015 the oligomannosylated glycopeptides 981:1015 the oligomannosylated glycopeptides with the same Y1 ion 981:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 0 24 theme N-Glycosylation 97:111 arg1 Characterization 63:78 Accurate Characterization 54:78 Accurate Characterization of Site-Specific N-Glycosylation 54:111 N-Glycopeptide Reduction with Exoglycosidases Enables Accurate Characterization of Site-Specific N-Glycosylation. 27808502 6 25 theme other 1252:1256 arg1 samples 1258:1264 other samples 1252:1264 other samples 1252:1264 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 7 26 from lysates 1548:1554 arg1 identification 1487:1500 the identification 1483:1500 the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1483:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 26 from lysates 1548:1554 arg1 N-glycopeptides 1509:1523 609 N-glycopeptides 1505:1523 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1505:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 4 27 gly glycopeptides 1003:1015 arg2 glycopeptides 1003:1015 the oligomannosylated glycopeptides 981:1015 the oligomannosylated glycopeptides with the same Y1 ion 981:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 0 28 theme N-Glycopeptide 0:13 arg1 Reduction 15:23 N-Glycopeptide Reduction 0:23 N-Glycopeptide Reduction with Exoglycosidases 0:44 N-Glycopeptide Reduction with Exoglycosidases Enables Accurate Characterization of Site-Specific N-Glycosylation. 27808502 2 29 theme glycopeptides 384:396 arg1 dependent 418:426 dependent 418:426 dependent 418:426 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 2 29 theme glycopeptides 384:396 arg1 fragmentation 367:379 the fragmentation 363:379 the fragmentation of glycopeptides 363:396 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 7 30 theme glycopeptidome 1459:1472 arg1 analysis 1384:1391 simultaneous analysis 1371:1391 simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome 1371:1472 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 6 31 theme site-specific 1297:1309 arg1 N-glycosylation 1311:1325 site-specific N-glycosylation 1297:1325 site-specific N-glycosylation involving complex N-glycans 1297:1353 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 6 32 theme complex 1337:1343 arg1 N-glycans 1345:1353 complex N-glycans 1337:1353 complex N-glycans 1337:1353 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 1 33 theme new 126:128 arg1 approach 130:137 a new approach 124:137 a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases 124:315 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 7 34 gly oligomannosylated 1409:1425 arg1 N-glycopeptidome 1427:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 2 35 theme ions 513:516 arg1 generation 496:505 the generation 492:505 the generation of Y1 ions 492:516 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 7 36 theme Huh7 1559:1562 arg1 cells 1564:1568 Huh7 cells 1559:1568 Huh7 cells 1559:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 3 37 theme site 772:775 arg1 identification 722:735 the identification 718:735 the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides 718:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 38 gly Oligomannosylated 564:580 arg1 glycopeptidome 582:595 Oligomannosylated glycopeptidome 564:595 Oligomannosylated glycopeptidome produced by NGlycoReduction 564:623 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 1 39 theme N-glycopeptide 260:273 arg1 core 291:294 the N-glycopeptide pentasaccharide core 256:294 the N-glycopeptide pentasaccharide core 256:294 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 0 40 theme Accurate 54:61 arg1 Characterization 63:78 Accurate Characterization 54:78 Accurate Characterization of Site-Specific N-Glycosylation 54:111 N-Glycopeptide Reduction with Exoglycosidases Enables Accurate Characterization of Site-Specific N-Glycosylation. 27808502 5 41 from confidence 1126:1135 arg1 alignment 1151:1159 the Y1 ion alignment 1140:1159 the Y1 ion alignment 1140:1159 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 7 42 from identification 1487:1500 arg1 lysates 1548:1554 the secretome and lysates 1530:1554 lysates 1548:1554 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 42 from identification 1487:1500 arg1 secretome 1534:1542 the secretome and lysates 1530:1554 secretome 1534:1542 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 43 theme oligomannosylated 1409:1425 arg1 N-glycopeptidome 1427:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 44 from secretome 1534:1542 arg1 identification 1487:1500 the identification 1483:1500 the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1483:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 44 from secretome 1534:1542 arg1 N-glycopeptides 1509:1523 609 N-glycopeptides 1505:1523 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1505:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 1 45 theme pentasaccharide 275:289 arg1 core 291:294 the N-glycopeptide pentasaccharide core 256:294 the N-glycopeptide pentasaccharide core 256:294 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 4 46 gly glycopeptides 851:863 arg2 glycopeptides 851:863 intact glycopeptides 844:863 intact glycopeptides 844:863 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 2 47 with glycans 432:438 arg1 structures 458:467 core mannose structures 445:467 core mannose structures 445:467 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 7 48 theme N-glycopeptidome 1427:1442 arg1 analysis 1384:1391 simultaneous analysis 1371:1391 simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome 1371:1472 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 2 49 theme mass 545:548 arg1 spectrometry 550:561 mass spectrometry 545:561 mass spectrometry 545:561 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 1 50 gly oligomannosylated 180:196 arg1 N-glycopeptidome 198:213 an oligomannosylated N-glycopeptidome 177:213 an oligomannosylated N-glycopeptidome 177:213 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 3 51 theme glycosylation 758:770 arg1 site 772:775 glycosylation site 758:775 glycosylation site 758:775 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 51 theme glycosylation 758:770 arg1 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 5 52 theme Y1 1144:1145 arg1 alignment 1151:1159 the Y1 ion alignment 1140:1159 the Y1 ion alignment 1140:1159 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 6 53 theme ease 1210:1213 arg1 advantage 1184:1192 the advantage 1180:1192 the advantage of low cost and ease of processing 1180:1227 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 7 54 gly N-glycopeptides 1509:1523 arg1 lysates 1548:1554 the secretome and lysates 1530:1554 lysates 1548:1554 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 54 gly N-glycopeptides 1509:1523 arg2 N-glycopeptides 1509:1523 609 N-glycopeptides 1505:1523 609 N-glycopeptides from the secretome and lysates of Huh7 cells 1505:1568 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 7 54 gly N-glycopeptides 1509:1523 arg1 secretome 1534:1542 the secretome and lysates 1530:1554 secretome 1534:1542 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 4 55 theme same 1026:1029 arg1 ion 1034:1036 the same Y1 ion 1022:1036 the same Y1 ion 1022:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 5 56 theme ion 1147:1149 arg1 alignment 1151:1159 the Y1 ion alignment 1140:1159 the Y1 ion alignment 1140:1159 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 0 57 with Reduction 15:23 arg1 Exoglycosidases 30:44 Exoglycosidases 30:44 Exoglycosidases 30:44 N-Glycopeptide Reduction with Exoglycosidases Enables Accurate Characterization of Site-Specific N-Glycosylation. 27808502 7 58 theme combined 1400:1407 arg1 N-glycopeptidome 1427:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 the combined oligomannosylated N-glycopeptidome 1396:1442 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 3 59 gly glycopeptides 805:817 arg2 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 60 theme intact 798:803 arg1 glycopeptides 805:817 intact glycopeptides 798:817 intact glycopeptides 798:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 3 61 theme peptide 740:746 arg1 sequence 748:755 peptide sequence 740:755 peptide sequence 740:755 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 7 62 theme simultaneous 1371:1382 arg1 analysis 1384:1391 simultaneous analysis 1371:1391 simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome 1371:1472 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 3 63 theme Oligomannosylated 564:580 arg1 glycopeptidome 582:595 Oligomannosylated glycopeptidome 564:595 Oligomannosylated glycopeptidome produced by NGlycoReduction 564:623 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 4 64 with glycopeptides 1003:1015 arg1 ion 1034:1036 the same Y1 ion 1022:1036 the same Y1 ion 1022:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 4 65 theme Y1 1031:1032 arg1 ion 1034:1036 the same Y1 ion 1022:1036 the same Y1 ion 1022:1036 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 3 66 theme sequence 748:755 arg1 identification 722:735 the identification 718:735 the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides 718:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 1 67 theme oligomannosylated 180:196 arg1 N-glycopeptidome 198:213 an oligomannosylated N-glycopeptidome 177:213 an oligomannosylated N-glycopeptidome 177:213 We report a new approach called NGlycoReduction that generates an oligomannosylated N-glycopeptidome by enzymatically removing sugars outside the N-glycopeptide pentasaccharide core with exoglycosidases. 27808502 7 68 theme native 1452:1457 arg1 glycopeptidome 1459:1472 the native glycopeptidome 1448:1472 the native glycopeptidome 1448:1472 In this study, simultaneous analysis of the combined oligomannosylated N-glycopeptidome and the native glycopeptidome leads to the identification of 609 N-glycopeptides from the secretome and lysates of Huh7 cells. 27808502 4 69 theme glycopeptides 851:863 arg1 structure 831:839 The glycan structure 820:839 The glycan structure of intact glycopeptides 820:863 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 2 70 gly glycopeptides 384:396 arg2 glycopeptides 384:396 glycopeptides 384:396 glycopeptides 384:396 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 3 71 theme structure 785:793 arg1 identification 722:735 the identification 718:735 the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides 718:817 Oligomannosylated glycopeptidome produced by NGlycoReduction can be mixed with the intact N-glycopeptidome and analyzed by HPLC-ESI-MS together to enable the identification of peptide sequence, glycosylation site and the structure of intact glycopeptides. 27808502 4 72 theme peptide 925:931 arg1 sequences 933:941 their peptide sequences 919:941 their peptide sequences 919:941 The glycan structure of intact glycopeptides can be identified from MS/MS spectra of their own and their peptide sequences were identified by the MS3 spectra of the oligomannosylated glycopeptides with the same Y1 ion. 27808502 5 73 used used 1105:1108 arg2 tolerance 1049:1057 mass tolerance 1044:1057 mass tolerance 1044:1057 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 5 73 used used 1105:1108 arg2 difference 1063:1072 difference 1063:1072 difference in retention time 1063:1090 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 2 74 dep discovery 348:356 arg1 lead 484:487 lead 484:487 lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry 484:561 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 2 74 dep discovery 348:356 arg1 fragmentation 367:379 the fragmentation 363:379 the fragmentation of glycopeptides 363:396 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 2 74 dep discovery 348:356 arg1 dependent 418:426 dependent 418:426 dependent 418:426 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27808502 5 75 from difference 1063:1072 arg1 time 1087:1090 retention time 1077:1090 retention time 1077:1090 Both mass tolerance and difference in retention time were further used to increase the confidence in the Y1 ion alignment. 27808502 0 76 theme Site-Specific 83:95 arg1 N-Glycosylation 97:111 Site-Specific N-Glycosylation 83:111 Site-Specific N-Glycosylation 83:111 N-Glycopeptide Reduction with Exoglycosidases Enables Accurate Characterization of Site-Specific N-Glycosylation. 27808502 6 77 contain has 1176:1178 arg2 advantage 1184:1192 the advantage 1180:1192 the advantage of low cost and ease of processing 1180:1227 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 6 77 contain has 1176:1178 arg1 approach 1167:1174 This approach 1162:1174 This approach 1162:1174 This approach has the advantage of low cost and ease of processing and can be expanded to other samples, especially for characterizing site-specific N-glycosylation involving complex N-glycans. 27808502 2 78 theme mannose 450:456 arg1 structures 458:467 core mannose structures 445:467 core mannose structures 445:467 This approach is based on our discovery that the fragmentation of glycopeptides is glycan-structure dependent and glycans with core mannose structures overwhelmingly lead to the generation of Y1 ions when subjected to MS/MS in mass spectrometry. 27550041 5 0 gly fucosylated 1047:1057 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 0 gly fucosylated 1047:1057 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 1 1 theme high-performance 357:372 arg1 chromatography 381:394 reversed-phase high-performance liquid chromatography 342:394 reversed-phase high-performance liquid chromatography 342:394 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 1 2 theme lactoferrin 181:191 arg1 N-glycans 156:164 the N-glycans 152:164 the N-glycans of donkey milk lactoferrin 152:191 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 3 3 theme neutral 843:849 arg1 most 779:782 most 779:782 most 779:782 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 27550041 3 3 theme neutral 843:849 arg1 complex/hybrid 851:864 neutral complex/hybrid 843:864 neutral complex/hybrid 843:864 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 27550041 3 3 theme neutral 843:849 arg1 N-glycans 791:799 the N-glycans 787:799 the N-glycans identified in donkey milk lactoferrin 787:837 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 27550041 3 4 theme N-glycan 734:741 arg1 structures 743:752 the N-glycan structures 730:752 the N-glycan structures determined 730:763 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 27550041 1 5 theme liquid 374:379 arg1 chromatography 381:394 reversed-phase high-performance liquid chromatography 342:394 reversed-phase high-performance liquid chromatography 342:394 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 6 6 theme goat 1336:1339 arg1 milk 1341:1344 goat milk 1336:1344 goat milk 1336:1344 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 6 7 dep human 1318:1322 arg1 lactoferrin 1346:1356 lactoferrin 1346:1356 lactoferrin 1346:1356 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 2 8 theme hybrid 585:590 arg1 N-glycans 592:600 hybrid N-glycans 585:600 hybrid N-glycans 585:600 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 8 theme hybrid 585:590 arg1 mannose 564:570 high mannose 559:570 high mannose 559:570 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 8 theme hybrid 585:590 arg1 structures 537:546 26 different glycan structures 517:546 26 different glycan structures 517:546 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 9 theme amide 641:645 arg1 bond 647:650 an amide bond 638:650 an amide bond to asparagine residues located at the positions 137, 281 and 476 638:715 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 6 10 theme monosaccharide 1223:1236 arg1 composition 1238:1248 the monosaccharide composition 1219:1248 the monosaccharide composition of the N-glycans of donkey milk lactoferrin 1219:1292 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 4 11 theme neutral 933:939 arg1 N-glycans 968:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 6 12 theme N-glycans 1257:1265 arg1 composition 1238:1248 the monosaccharide composition 1219:1248 the monosaccharide composition of the N-glycans of donkey milk lactoferrin 1219:1292 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 1 13 theme electrospray 397:408 arg1 spectrometry 415:426 electrospray mass spectrometry 397:426 electrospray mass spectrometry 397:426 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 4 14 theme non-fucosylated 886:900 arg1 N-glycans 917:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 5 15 theme sialylated 1036:1045 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 15 theme sialylated 1036:1045 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 1 16 theme mass 410:413 arg1 spectrometry 415:426 electrospray mass spectrometry 397:426 electrospray mass spectrometry 397:426 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 2 17 theme located 675:681 arg1 residues 666:673 asparagine residues 655:673 asparagine residues located at the positions 137, 281 and 476 655:715 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 5 18 gly sialylated 1085:1094 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 18 gly sialylated 1085:1094 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 19 gly non-fucosylated 1096:1110 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 19 gly non-fucosylated 1096:1110 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 20 gly sialylated 1036:1045 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 20 gly sialylated 1036:1045 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 6 21 theme composition 1238:1248 arg1 comparison 1205:1214 A comparison 1203:1214 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin 1203:1356 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 1 22 theme ion 206:208 arg1 chromatography 219:232 ion exchange chromatography 206:232 ion exchange chromatography 206:232 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 0 23 theme Site-specific 0:12 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of donkey milk lactoferrin 0:53 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 1 24 theme chymotryptic 291:302 arg1 digestion 304:312 chymotryptic digestion 291:312 chymotryptic digestion 291:312 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 2 25 theme asparagine 655:664 arg1 residues 666:673 asparagine residues 655:673 asparagine residues located at the positions 137, 281 and 476 655:715 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 1 26 theme comprehensive 108:120 arg1 composition 137:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 2 27 attach linked 603:608 arg2 mannose 564:570 high mannose 559:570 high mannose 559:570 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 27 attach linked 603:608 arg2 structures 537:546 26 different glycan structures 517:546 26 different glycan structures 517:546 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 27 attach linked 603:608 arg2 N-glycans 592:600 hybrid N-glycans 585:600 hybrid N-glycans 585:600 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 27 attach linked 603:608 arg2 complex 573:579 complex 573:579 complex 573:579 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 27 attach linked 603:608 arg1 backbone 625:632 the protein backbone 613:632 the protein backbone 613:632 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 27 attach linked 603:608 arg3 bond 647:650 an amide bond 638:650 an amide bond to asparagine residues located at the positions 137, 281 and 476 638:715 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 1 28 theme exchange 210:217 arg1 chromatography 219:232 ion exchange chromatography 206:232 ion exchange chromatography 206:232 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 0 29 theme donkey 31:36 arg1 lactoferrin 43:53 donkey milk lactoferrin 31:53 donkey milk lactoferrin 31:53 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 7 30 with ProteomeXchange 1394:1408 arg1 identifier 1415:1424 identifier PXD004289 1415:1434 identifier PXD004289 1415:1434 Data are available via ProteomeXchange with identifier PXD004289. 27550041 1 31 theme monosaccharide 122:135 arg1 composition 137:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 5 32 theme high 1007:1010 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 32 theme high 1007:1010 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 32 theme high 1007:1010 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 33 theme N-glycans 1020:1028 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 33 theme N-glycans 1020:1028 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 33 theme N-glycans 1020:1028 arg1 one 1131:1133 one 1131:1133 one 1131:1133 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 33 theme N-glycans 1020:1028 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 4 34 gly non-fucosylated 886:900 arg1 N-glycans 917:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 0 35 theme lactoferrin 43:53 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of donkey milk lactoferrin 0:53 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 5 36 located found 1196:1200 arg2 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 36 located found 1196:1200 arg2 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 36 located found 1196:1200 arg1 addition 993:1000 addition 993:1000 addition 993:1000 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 36 located found 1196:1200 arg2 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 1 37 attach isolated 194:201 arg2 composition 137:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 1 37 attach isolated 194:201 arg1 sample 258:263 an individual milk sample 239:263 an individual milk sample 239:263 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 5 38 theme sialylated 1085:1094 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 38 theme sialylated 1085:1094 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 4 39 gly fucosylated 941:951 arg1 N-glycans 968:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 0 40 theme milk 38:41 arg1 lactoferrin 43:53 donkey milk lactoferrin 31:53 donkey milk lactoferrin 31:53 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 3 41 theme donkey 815:820 arg1 lactoferrin 827:837 donkey milk lactoferrin 815:837 donkey milk lactoferrin 815:837 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 27550041 2 42 theme glycan 530:535 arg1 complex 573:579 complex 573:579 complex 573:579 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 42 theme glycan 530:535 arg1 N-glycans 592:600 hybrid N-glycans 585:600 hybrid N-glycans 585:600 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 42 theme glycan 530:535 arg1 mannose 564:570 high mannose 559:570 high mannose 559:570 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 42 theme glycan 530:535 arg1 structures 537:546 26 different glycan structures 517:546 26 different glycan structures 517:546 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 1 43 theme high 433:436 arg1 fragmentation 461:473 high collision dissociation fragmentation 433:473 high collision dissociation fragmentation 433:473 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 0 44 gly glycosylation 14:26 arg1 lactoferrin 43:53 donkey milk lactoferrin 31:53 donkey milk lactoferrin 31:53 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 1 45 theme individual 242:251 arg1 sample 258:263 an individual milk sample 239:263 an individual milk sample 239:263 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 2 46 theme different 520:528 arg1 complex 573:579 complex 573:579 complex 573:579 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 46 theme different 520:528 arg1 N-glycans 592:600 hybrid N-glycans 585:600 hybrid N-glycans 585:600 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 46 theme different 520:528 arg1 mannose 564:570 high mannose 559:570 high mannose 559:570 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 46 theme different 520:528 arg1 structures 537:546 26 different glycan structures 517:546 26 different glycan structures 517:546 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 1 47 theme HILIC 324:328 arg1 enrichment 330:339 HILIC enrichment 324:339 HILIC enrichment 324:339 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 1 48 theme collision 438:446 arg1 fragmentation 461:473 high collision dissociation fragmentation 433:473 high collision dissociation fragmentation 433:473 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 5 49 theme complex 1112:1118 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 49 theme complex 1112:1118 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 4 50 theme complex/hybrid 902:915 arg1 N-glycans 917:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 2 51 from positions 690:698 arg1 located 675:681 located 675:681 located 675:681 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 4 52 theme complex/hybrid 953:966 arg1 N-glycans 968:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 1 53 theme dissociation 448:459 arg1 fragmentation 461:473 high collision dissociation fragmentation 433:473 high collision dissociation fragmentation 433:473 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 4 54 theme neutral 878:884 arg1 N-glycans 917:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 10 neutral non-fucosylated complex/hybrid N-glycans 875:925 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 4 55 theme fucosylated 941:951 arg1 N-glycans 968:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 4 neutral fucosylated complex/hybrid N-glycans 931:976 Indeed, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. 27550041 2 56 theme protein 617:623 arg1 backbone 625:632 the protein backbone 613:632 the protein backbone 613:632 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 5 57 theme complex 1059:1065 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 57 theme complex 1059:1065 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 58 theme non-fucosylated 1096:1110 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 58 theme non-fucosylated 1096:1110 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 1 59 theme N-glycans 156:164 arg1 composition 137:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition 106:147 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 5 60 theme mannose 1012:1018 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 60 theme mannose 1012:1018 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 60 theme mannose 1012:1018 arg1 N-glycans 1120:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 six sialylated non-fucosylated complex N-glycans 1081:1128 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 1 61 theme milk 253:256 arg1 sample 258:263 an individual milk sample 239:263 an individual milk sample 239:263 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 5 62 theme fucosylated 1047:1057 arg1 N-glycans 1020:1028 two high mannose N-glycans 1003:1028 two high mannose N-glycans 1003:1028 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 62 theme fucosylated 1047:1057 arg1 N-glycans 1067:1075 four sialylated fucosylated complex N-glycans 1031:1075 four sialylated fucosylated complex N-glycans 1031:1075 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 6 63 theme lactoferrin 1282:1292 arg1 N-glycans 1257:1265 the N-glycans 1253:1265 the N-glycans of donkey milk lactoferrin 1253:1292 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 5 64 theme N-glycolylneuraminic 1155:1174 arg1 Neu5Gc 1182:1187 Neu5Gc 1182:1187 Neu5Gc 1182:1187 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 5 64 theme N-glycolylneuraminic 1155:1174 arg1 acid 1176:1179 N-glycolylneuraminic acid 1155:1179 N-glycolylneuraminic acid (Neu5Gc) 1155:1188 In addition, two high mannose N-glycans, four sialylated fucosylated complex N-glycans and six sialylated non-fucosylated complex N-glycans, one of which containing N-glycolylneuraminic acid (Neu5Gc), were found. 27550041 2 65 theme high 559:562 arg1 mannose 564:570 high mannose 559:570 high mannose 559:570 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 0 66 theme mass 87:90 arg1 spectrometry 92:103 high-resolution mass spectrometry 71:103 high-resolution mass spectrometry 71:103 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 6 67 theme milk 1277:1280 arg1 lactoferrin 1282:1292 donkey milk lactoferrin 1270:1292 donkey milk lactoferrin 1270:1292 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 1 68 theme donkey 169:174 arg1 lactoferrin 181:191 donkey milk lactoferrin 169:191 donkey milk lactoferrin 169:191 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 0 69 theme high-resolution 71:85 arg1 spectrometry 92:103 high-resolution mass spectrometry 71:103 high-resolution mass spectrometry 71:103 Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry. 27550041 1 70 theme reversed-phase 342:355 arg1 chromatography 381:394 reversed-phase high-performance liquid chromatography 342:394 reversed-phase high-performance liquid chromatography 342:394 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 6 71 theme donkey 1270:1275 arg1 lactoferrin 1282:1292 donkey milk lactoferrin 1270:1292 donkey milk lactoferrin 1270:1292 A comparison of the monosaccharide composition of the N-glycans of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported. 27550041 1 72 theme milk 176:179 arg1 lactoferrin 181:191 donkey milk lactoferrin 169:191 donkey milk lactoferrin 169:191 A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. 27550041 2 73 dep positions 690:698 arg1 137 700:702 137 700:702 137 700:702 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 73 dep positions 690:698 arg1 476 713:715 476 713:715 476 713:715 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 2 73 dep positions 690:698 arg1 281 705:707 281 705:707 281 705:707 The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27550041 3 74 theme milk 822:825 arg1 lactoferrin 827:837 donkey milk lactoferrin 815:837 donkey milk lactoferrin 815:837 Altogether, the N-glycan structures determined revealed that most of the N-glycans identified in donkey milk lactoferrin are neutral complex/hybrid. 28680094 3 0 theme αIIbβ3 310:315 arg1 structures 296:305 Crystal structures 288:305 Crystal structures of αIIbβ3 and αVβ3 288:324 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 5 1 theme N-glycan 654:661 arg1 site 663:666 the N-glycan site 650:666 the N-glycan site 650:666 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 5 1 theme N-glycan 654:661 arg1 β3-N320 669:675 β3-N320 669:675 β3-N320 669:675 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 7 2 attach removal 1002:1008 arg2 N-glycan 1025:1032 the β3-N371 N-glycan 1013:1032 the β3-N371 N-glycan 1013:1032 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 7 2 attach removal 1002:1008 arg3 domain 1113:1118 the I-EGF1 domain 1102:1118 the I-EGF1 domain 1102:1118 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 2 3 located found 243:247 arg2 variations 167:176 Great variations 161:176 Great variations in both the number and distribution of N-glycosylation sites 161:237 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 2 3 located found 243:247 arg1 subunits 278:285 the 18 α and 8 β integrin subunits 252:285 the 18 α and 8 β integrin subunits 252:285 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 0 4 theme conformational 68:81 arg1 regulation 83:92 conformational regulation 68:92 conformational regulation 68:92 The importance of N-glycosylation on β3 integrin ligand binding and conformational regulation. 28680094 7 5 dep active 1146:1151 arg1 integrin 1132:1139 β3 integrin 1129:1139 β3 integrin 1129:1139 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 6 6 theme domain 826:831 arg1 interface 833:841 the β3-I-EGF3 and αIIb-calf-1 domain interface 796:841 interface 833:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 4 7 theme individual 583:592 arg1 sites 603:607 individual N-glycan sites 583:607 individual N-glycan sites 583:607 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 3 8 theme precise 344:350 arg1 location 363:370 the precise structural location 340:370 the precise structural location of each N-glycan site 340:392 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 6 9 theme αIIb-calf-1 814:824 arg1 interface 833:841 the β3-I-EGF3 and αIIb-calf-1 domain interface 796:841 interface 833:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 2 10 dep number 190:195 arg1 the 186:188 the 186:188 the 186:188 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 3 11 theme structural 352:361 arg1 location 363:370 the precise structural location 340:370 the precise structural location of each N-glycan site 340:392 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 6 12 theme domain 902:907 arg1 interface 909:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 8 13 from domain 1217:1222 arg1 N-glycan 1198:1205 one unique N-glycan 1187:1205 one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation 1187:1278 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 2 14 theme integrin 269:276 arg1 subunits 278:285 the 18 α and 8 β integrin subunits 252:285 the 18 α and 8 β integrin subunits 252:285 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 4 15 theme sites 603:607 arg1 function 571:578 the function 567:578 the function of individual N-glycan sites in β3 integrin activation 567:633 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 0 16 theme ligand 49:54 arg1 binding 56:62 integrin ligand binding 40:62 integrin ligand binding 40:62 The importance of N-glycosylation on β3 integrin ligand binding and conformational regulation. 28680094 2 17 theme Great 161:165 arg1 variations 167:176 Great variations 161:176 Great variations in both the number and distribution of N-glycosylation sites 161:237 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 7 18 theme β3-N452 1082:1088 arg1 N-glycan 1090:1097 the β3-N452 N-glycan 1078:1097 the β3-N452 N-glycan 1078:1097 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 8 19 theme β1 1227:1228 arg1 subunit 1230:1236 β1 subunit 1227:1236 β1 subunit 1227:1236 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 2 20 theme β 267:267 arg1 subunits 278:285 the 18 α and 8 β integrin subunits 252:285 the 18 α and 8 β integrin subunits 252:285 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 8 21 theme βI 1214:1215 arg1 domain 1217:1222 the βI domain 1210:1222 the βI domain of β1 subunit 1210:1236 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 8 21 theme βI 1214:1215 arg1 subunit 1230:1236 β1 subunit 1227:1236 β1 subunit 1227:1236 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 4 22 theme N-glycan 594:601 arg1 sites 603:607 individual N-glycan sites 583:607 individual N-glycan sites 583:607 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 4 23 from function 571:578 arg1 activation 624:633 β3 integrin activation 612:633 β3 integrin activation 612:633 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 3 24 theme individual 430:439 arg1 site 450:453 individual N-glycan site 430:453 individual N-glycan site 430:453 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 6 25 theme β3-β-tail 876:884 arg1 interface 909:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 5 26 theme αVβ3 755:758 arg1 activation 760:769 αVβ3 activation 755:769 αIIbβ3 but not αVβ3 activation 740:769 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 6 27 theme αIIb-calf-2 890:900 arg1 interface 909:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 the β3-β-tail and αIIb-calf-2 domain interface 872:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 3 28 theme structural 403:412 arg1 consequences 414:425 the structural consequences 399:425 the structural consequences of individual N-glycan site on integrin activation 399:476 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 3 29 theme N-glycan 380:387 arg1 site 389:392 each N-glycan site 375:392 each N-glycan site 375:392 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 9 30 theme domain 1428:1433 arg1 interfaces 1435:1444 the domain interfaces 1424:1444 the domain interfaces of integrin 1424:1456 Our study suggests that the bulky N-glycans influence the large-scale conformational rearrangement by potentially stabilizing or destabilizing the domain interfaces of integrin. 28680094 9 31 theme integrin 1449:1456 arg1 interfaces 1435:1444 the domain interfaces 1424:1444 the domain interfaces of integrin 1424:1456 Our study suggests that the bulky N-glycans influence the large-scale conformational rearrangement by potentially stabilizing or destabilizing the domain interfaces of integrin. 28680094 0 32 theme N-glycosylation 18:32 arg1 importance 4:13 The importance 0:13 The importance of N-glycosylation on 0:35 The importance of N-glycosylation on β3 integrin ligand binding and conformational regulation. 28680094 6 33 theme integrins 978:986 arg1 activation 943:952 the activation 939:952 the activation of both αIIbβ3 and αVβ3 integrins 939:986 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 5 34 theme leg 698:700 arg1 interface 709:717 the headpiece and leg domain interface 680:717 interface 709:717 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 6 35 theme β3-N559 776:782 arg1 N-glycan 784:791 The β3-N559 N-glycan 772:791 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface 772:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 8 36 theme subunit 1230:1236 arg1 domain 1217:1222 the βI domain 1210:1222 the βI domain of β1 subunit 1210:1236 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 8 36 theme subunit 1230:1236 arg1 subunit 1230:1236 β1 subunit 1227:1236 β1 subunit 1227:1236 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 6 37 theme αVβ3 973:976 arg1 integrins 978:986 both αIIbβ3 and αVβ3 integrins 957:986 both αIIbβ3 and αVβ3 integrins 957:986 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 5 38 theme domain 702:707 arg1 interface 709:717 the headpiece and leg domain interface 680:717 interface 709:717 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 3 39 theme integrin 458:465 arg1 activation 467:476 integrin activation 458:476 integrin activation 458:476 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 8 40 theme unique 1191:1196 arg1 N-glycan 1198:1205 one unique N-glycan 1187:1205 one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation 1187:1278 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 7 41 theme wild 1162:1165 arg1 type 1167:1170 the wild type 1158:1170 the wild type 1158:1170 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 3 42 theme N-glycan 441:448 arg1 site 450:453 individual N-glycan site 430:453 individual N-glycan site 430:453 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 6 43 from interface 833:841 arg1 N-glycan 860:867 the β3-N654 N-glycan 848:867 the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface 848:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 6 43 from interface 833:841 arg1 N-glycan 784:791 The β3-N559 N-glycan 772:791 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface 772:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 6 44 theme αIIbβ3 962:967 arg1 integrins 978:986 both αIIbβ3 and αVβ3 integrins 957:986 both αIIbβ3 and αVβ3 integrins 957:986 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 7 45 theme I-EGF3 1057:1062 arg1 interface 1064:1072 I-EGF3 interface 1057:1072 I-EGF3 interface 1057:1072 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 9 46 theme bulky 1309:1313 arg1 N-glycans 1315:1323 the bulky N-glycans 1305:1323 the bulky N-glycans 1305:1323 Our study suggests that the bulky N-glycans influence the large-scale conformational rearrangement by potentially stabilizing or destabilizing the domain interfaces of integrin. 28680094 5 47 from interface 709:717 arg1 site 663:666 the N-glycan site 650:666 the N-glycan site 650:666 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 5 47 from interface 709:717 arg1 β3-N320 669:675 β3-N320 669:675 β3-N320 669:675 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 3 48 theme site 450:453 arg1 consequences 414:425 the structural consequences 399:425 the structural consequences of individual N-glycan site on integrin activation 399:476 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 3 49 theme site 389:392 arg1 location 363:370 the precise structural location 340:370 the precise structural location of each N-glycan site 340:392 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 4 50 theme structure-guided 527:542 arg1 analyses 544:551 structure-guided analyses 527:551 structure-guided analyses 527:551 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 7 51 theme β3 1043:1044 arg1 hybrid 1046:1051 the β3 hybrid 1039:1051 the β3 hybrid 1039:1051 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 6 52 theme β3-N654 852:858 arg1 N-glycan 860:867 the β3-N654 N-glycan 848:867 the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface 848:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 7 53 theme N-glycan 1025:1032 arg1 removal 1002:1008 removal 1002:1008 removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain 1002:1118 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 8 54 theme α5β1 1264:1267 arg1 activation 1269:1278 α5β1 activation 1264:1278 α5β1 activation 1264:1278 We identified one unique N-glycan at the βI domain of β1 subunit that negatively regulates α5β1 activation. 28680094 0 55 theme integrin 40:47 arg1 binding 56:62 integrin ligand binding 40:62 integrin ligand binding 40:62 The importance of N-glycosylation on β3 integrin ligand binding and conformational regulation. 28680094 9 56 theme conformational 1351:1364 arg1 rearrangement 1366:1378 the large-scale conformational rearrangement 1335:1378 the large-scale conformational rearrangement 1335:1378 Our study suggests that the bulky N-glycans influence the large-scale conformational rearrangement by potentially stabilizing or destabilizing the domain interfaces of integrin. 28680094 2 57 theme N-glycosylation 217:231 arg1 sites 233:237 N-glycosylation sites 217:237 N-glycosylation sites 217:237 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 2 58 theme sites 233:237 arg1 distribution 201:212 distribution 201:212 distribution 201:212 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 2 58 theme sites 233:237 arg1 number 190:195 number 190:195 number 190:195 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 1 59 theme adhesive 129:136 arg1 function 138:145 the adhesive function 125:145 the adhesive function of integrins 125:158 N-glycosylations can regulate the adhesive function of integrins. 28680094 6 60 from interface 909:917 arg1 N-glycan 860:867 the β3-N654 N-glycan 848:867 the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface 848:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 6 60 from interface 909:917 arg1 N-glycan 784:791 The β3-N559 N-glycan 772:791 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface 772:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 2 61 gly N-glycosylation 217:231 arg2 sites 233:237 N-glycosylation sites 217:237 N-glycosylation sites 217:237 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 4 62 theme integrin 615:622 arg1 activation 624:633 β3 integrin activation 612:633 β3 integrin activation 612:633 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 3 63 theme Crystal 288:294 arg1 structures 296:305 Crystal structures 288:305 Crystal structures of αIIbβ3 and αVβ3 288:324 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 7 64 theme I-EGF1 1106:1111 arg1 domain 1113:1118 the I-EGF1 domain 1102:1118 the I-EGF1 domain 1102:1118 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 4 65 theme β3 612:613 arg1 activation 624:633 β3 integrin activation 612:633 β3 integrin activation 612:633 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 3 66 theme αVβ3 321:324 arg1 structures 296:305 Crystal structures 288:305 Crystal structures of αIIbβ3 and αVβ3 288:324 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 3 67 from consequences 414:425 arg1 activation 467:476 integrin activation 458:476 integrin activation 458:476 Crystal structures of αIIbβ3 and αVβ3 have resolved the precise structural location of each N-glycan site, but the structural consequences of individual N-glycan site on integrin activation remain unclear. 28680094 1 68 theme integrins 150:158 arg1 function 138:145 the adhesive function 125:145 the adhesive function of integrins 125:158 N-glycosylations can regulate the adhesive function of integrins. 28680094 6 69 from β3-I-EGF3 800:808 arg1 N-glycan 860:867 the β3-N654 N-glycan 848:867 the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface 848:917 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 6 69 from β3-I-EGF3 800:808 arg1 N-glycan 784:791 The β3-N559 N-glycan 772:791 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface 772:841 The β3-N559 N-glycan at the β3-I-EGF3 and αIIb-calf-1 domain interface, and the β3-N654 N-glycan at the β3-β-tail and αIIb-calf-2 domain interface positively regulate the activation of both αIIbβ3 and αVβ3 integrins. 28680094 2 70 from variations 167:176 arg1 distribution 201:212 distribution 201:212 distribution 201:212 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 2 70 from variations 167:176 arg1 number 190:195 number 190:195 number 190:195 Great variations in both the number and distribution of N-glycosylation sites are found in the 18 α and 8 β integrin subunits. 28680094 4 71 theme site-directed 497:509 arg1 mutagenesis 511:521 site-directed mutagenesis 497:521 site-directed mutagenesis 497:521 By site-directed mutagenesis and structure-guided analyses, we dissected the function of individual N-glycan sites in β3 integrin activation. 28680094 7 72 from domain 1113:1118 arg1 removal 1002:1008 removal 1002:1008 removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain 1002:1118 In contrast, removal of the β3-N371 N-glycan near the β3 hybrid and I-EGF3 interface, or the β3-N452 N-glycan at the I-EGF1 domain rendered β3 integrin more active than the wild type. 28680094 5 73 from headpiece 684:692 arg1 site 663:666 the N-glycan site 650:666 the N-glycan site 650:666 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 5 73 from headpiece 684:692 arg1 β3-N320 669:675 β3-N320 669:675 β3-N320 669:675 We found that the N-glycan site, β3-N320 at the headpiece and leg domain interface positively regulates αIIbβ3 but not αVβ3 activation. 28680094 9 74 theme large-scale 1339:1349 arg1 rearrangement 1366:1378 the large-scale conformational rearrangement 1335:1378 the large-scale conformational rearrangement 1335:1378 Our study suggests that the bulky N-glycans influence the large-scale conformational rearrangement by potentially stabilizing or destabilizing the domain interfaces of integrin. 27161092 8 0 theme fucosylation 1345:1356 arg1 loss 1317:1320 loss 1317:1320 loss of mucin FUT2-mediated fucosylation 1317:1356 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 5 1 theme mice 841:844 arg1 models 846:851 glycoengineered mice models 825:851 glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype 825:930 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 3 2 theme present 517:523 arg1 glycans 509:515 host glycans 504:515 host glycans present in the glycocalyx of epithelial cells and extracellular mucus 504:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 4 3 theme MUC5AC 601:606 arg1 mucin 608:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 3 theme MUC5AC 601:606 arg1 component 626:634 the main component 617:634 the main component of the gastric mucus layer 617:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 6 4 theme Muc5ac 1014:1019 arg1 pattern 1003:1009 the O-glycosylation pattern 983:1009 the O-glycosylation pattern of Muc5ac 983:1019 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 3 5 from present 517:523 arg1 glycocalyx 532:541 the glycocalyx 528:541 the glycocalyx of epithelial cells and extracellular mucus 528:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 2 6 theme binding 349:355 arg1 barrier 283:289 This mucus barrier 272:289 This mucus barrier 272:289 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 2 6 theme binding 349:355 arg1 site 357:360 a binding site 347:360 a binding site that supports a sheltered niche of microbial adherence 347:415 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 1 7 theme mechanical 248:257 arg1 aggressions 259:269 biochemical and mechanical aggressions 232:269 biochemical and mechanical aggressions 232:269 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 8 8 theme BabA 1403:1406 arg1 adhesin 1408:1414 H. pylori BabA adhesin 1393:1414 H. pylori BabA adhesin 1393:1414 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 9 theme recognized 1428:1437 arg1 binding 1382:1388 gastric mucosal binding 1366:1388 gastric mucosal binding 1366:1388 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 9 theme recognized 1428:1437 arg1 feature 1439:1445 a recognized feature 1426:1445 a recognized feature of pathogenicity 1426:1462 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 10 theme mucosal 1374:1380 arg1 binding 1382:1388 gastric mucosal binding 1366:1388 gastric mucosal binding 1366:1388 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 10 theme mucosal 1374:1380 arg1 feature 1439:1445 a recognized feature 1426:1445 a recognized feature of pathogenicity 1426:1462 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 3 11 theme cells 557:561 arg1 glycocalyx 532:541 the glycocalyx 528:541 the glycocalyx of epithelial cells and extracellular mucus 528:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 6 12 theme Lewis-a 1167:1173 arg1 decrease 1062:1069 a marked decrease 1053:1069 a marked decrease in α1,2-fucosylated structures 1053:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 6 12 theme Lewis-a 1167:1173 arg1 expression 1116:1125 increased expression 1106:1125 increased expression of the terminal type 1 glycan structure Lewis-a 1106:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 1 13 theme thick 157:161 arg1 layer 175:179 a thick and complex layer 155:179 a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions 155:269 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 0 14 theme Helicobacter 87:98 arg1 binding 107:113 Helicobacter pylori binding 87:113 Helicobacter pylori binding 87:113 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 8 15 theme gastric 1366:1372 arg1 binding 1382:1388 gastric mucosal binding 1366:1388 gastric mucosal binding 1366:1388 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 15 theme gastric 1366:1372 arg1 feature 1439:1445 a recognized feature 1426:1445 a recognized feature of pathogenicity 1426:1462 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 7 16 theme murine 1264:1269 arg1 mucosa 1279:1284 murine gastric mucosa 1264:1284 murine gastric mucosa 1264:1284 Importantly, for the first time, we structurally validated the expression of Lewis-a in murine gastric mucosa. 27161092 5 17 theme non-secretor 903:914 arg1 phenotype 922:930 a non-secretor human phenotype 901:930 a non-secretor human phenotype 901:930 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 6 18 theme Fut2 1027:1030 arg1 knock-out 1032:1040 Fut2 knock-out 1027:1040 Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a 1027:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 4 19 theme pylori 696:701 arg1 binding 682:688 BabA-mediated binding 668:688 BabA-mediated binding of H. pylori to MUC5AC 668:711 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 6 20 theme structure 1157:1165 arg1 Lewis-a 1167:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 3 21 from glycocalyx 532:541 arg1 present 517:523 present 517:523 present 517:523 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 2 22 theme mucus 277:281 arg1 barrier 283:289 This mucus barrier 272:289 This mucus barrier 272:289 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 2 22 theme mucus 277:281 arg1 site 357:360 a binding site 347:360 a binding site that supports a sheltered niche of microbial adherence 347:415 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 3 23 theme carcinogenic 422:433 arg1 bacteria 435:442 The carcinogenic bacteria 418:442 The carcinogenic bacteria Helicobacter pylori 418:462 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 3 24 theme Helicobacter 444:455 arg1 bacteria 435:442 The carcinogenic bacteria 418:442 The carcinogenic bacteria Helicobacter pylori 418:462 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 5 25 theme human 916:920 arg1 phenotype 922:930 a non-secretor human phenotype 901:930 a non-secretor human phenotype 901:930 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 6 26 gly O-glycosylation 987:1001 arg1 Muc5ac 1014:1019 Muc5ac 1014:1019 Muc5ac 1014:1019 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 0 27 dep Helicobacter 87:98 arg1 pylori 100:105 pylori 100:105 pylori 100:105 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 1 28 theme complex 167:173 arg1 layer 175:179 a thick and complex layer 155:179 a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions 155:269 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 0 29 theme gastric 7:13 arg1 glycosylation 21:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 6 30 theme increased 1106:1114 arg1 expression 1116:1125 increased expression 1106:1125 increased expression of the terminal type 1 glycan structure Lewis-a 1106:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 7 31 theme gastric 1271:1277 arg1 mucosa 1279:1284 murine gastric mucosa 1264:1284 murine gastric mucosa 1264:1284 Importantly, for the first time, we structurally validated the expression of Lewis-a in murine gastric mucosa. 27161092 4 32 theme overt 740:744 arg1 disease 746:752 overt disease 740:752 overt disease 740:752 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 7 33 theme first 1197:1201 arg1 time 1203:1206 the first time 1193:1206 the first time 1193:1206 Importantly, for the first time, we structurally validated the expression of Lewis-a in murine gastric mucosa. 27161092 2 34 theme sheltered 378:386 arg1 niche 388:392 a sheltered niche 376:392 a sheltered niche of microbial adherence 376:415 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 0 35 theme Muc5ac 0:5 arg1 glycosylation 21:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 6 36 from expression 1116:1125 arg1 structures 1091:1100 α1,2-fucosylated structures 1074:1100 α1,2-fucosylated structures 1074:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 6 37 theme marked 1055:1060 arg1 decrease 1062:1069 a marked decrease 1053:1069 a marked decrease in α1,2-fucosylated structures 1053:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 8 38 theme H. 1393:1394 arg1 adhesin 1408:1414 H. pylori BabA adhesin 1393:1414 H. pylori BabA adhesin 1393:1414 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 4 39 theme BabA-mediated 668:680 arg1 binding 682:688 BabA-mediated binding 668:688 BabA-mediated binding of H. pylori to MUC5AC 668:711 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 5 40 theme Muc5ac 813:818 arg1 profile 802:808 the O-glycosylation profile 782:808 the O-glycosylation profile of Muc5ac 782:818 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 6 41 from decrease 1062:1069 arg1 structures 1091:1100 α1,2-fucosylated structures 1074:1100 α1,2-fucosylated structures 1074:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 3 42 theme extracellular 567:579 arg1 mucus 581:585 extracellular mucus 567:585 extracellular mucus 567:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 1 43 theme mucus 184:188 arg1 layer 175:179 a thick and complex layer 155:179 a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions 155:269 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 0 44 theme mucin 15:19 arg1 glycosylation 21:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation 0:33 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 8 45 theme pathogenicity 1450:1462 arg1 binding 1382:1388 gastric mucosal binding 1366:1388 gastric mucosal binding 1366:1388 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 45 theme pathogenicity 1450:1462 arg1 feature 1439:1445 a recognized feature 1426:1445 a recognized feature of pathogenicity 1426:1462 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 3 46 theme mucus 581:585 arg1 glycocalyx 532:541 the glycocalyx 528:541 the glycocalyx of epithelial cells and extracellular mucus 528:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 6 47 theme terminal 1134:1141 arg1 Lewis-a 1167:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 6 48 theme glycan 1150:1155 arg1 Lewis-a 1167:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 4 49 theme layer 657:661 arg1 mucin 608:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 49 theme layer 657:661 arg1 component 626:634 the main component 617:634 the main component of the gastric mucus layer 617:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 8 50 theme FUT2-mediated 1331:1343 arg1 fucosylation 1345:1356 mucin FUT2-mediated fucosylation 1325:1356 mucin FUT2-mediated fucosylation 1325:1356 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 6 51 theme α1,2-fucosylated 1074:1089 arg1 structures 1091:1100 α1,2-fucosylated structures 1074:1100 α1,2-fucosylated structures 1074:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 2 52 theme adherence 407:415 arg1 niche 388:392 a sheltered niche 376:392 a sheltered niche of microbial adherence 376:415 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 5 53 theme FUT2 865:868 arg1 enzyme 870:875 the FUT2 enzyme 861:875 the FUT2 enzyme 861:875 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 4 54 theme secreted 592:599 arg1 mucin 608:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 54 theme secreted 592:599 arg1 component 626:634 the main component 617:634 the main component of the gastric mucus layer 617:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 8 55 theme mucin 1325:1329 arg1 fucosylation 1345:1356 mucin FUT2-mediated fucosylation 1325:1356 mucin FUT2-mediated fucosylation 1325:1356 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 2 56 theme microbial 397:405 arg1 adherence 407:415 microbial adherence 397:415 microbial adherence 397:415 This mucus barrier confers protection against pathogens but also serves as a binding site that supports a sheltered niche of microbial adherence. 27161092 0 57 theme FUT2 48:51 arg1 activity 53:60 FUT2 activity 48:60 FUT2 activity 48:60 Muc5ac gastric mucin glycosylation is shaped by FUT2 activity and functionally impacts Helicobacter pylori binding. 27161092 8 58 theme pylori 1396:1401 arg1 adhesin 1408:1414 H. pylori BabA adhesin 1393:1414 H. pylori BabA adhesin 1393:1414 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 6 59 theme O-glycosylation 987:1001 arg1 pattern 1003:1009 the O-glycosylation pattern 983:1009 the O-glycosylation pattern of Muc5ac 983:1019 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 3 60 dep Helicobacter 444:455 arg1 pylori 457:462 pylori 457:462 pylori 457:462 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 3 61 theme host 504:507 arg1 glycans 509:515 host glycans 504:515 host glycans present in the glycocalyx of epithelial cells and extracellular mucus 504:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 1 62 theme gastrointestinal 120:135 arg1 tract 137:141 The gastrointestinal tract 116:141 The gastrointestinal tract 116:141 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 5 63 dep models 846:851 arg1 lacking 853:859 lacking 853:859 lacking the FUT2 enzyme 853:875 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 5 63 dep models 846:851 arg1 mimicking 891:899 mimicking 891:899 therefore mimicking a non-secretor human phenotype 881:930 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 6 64 gly α1,2-fucosylated 1074:1089 arg1 structures 1091:1100 α1,2-fucosylated structures 1074:1100 α1,2-fucosylated structures 1074:1100 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 7 65 from expression 1239:1248 arg1 mucosa 1279:1284 murine gastric mucosa 1264:1284 murine gastric mucosa 1264:1284 Importantly, for the first time, we structurally validated the expression of Lewis-a in murine gastric mucosa. 27161092 3 66 attach present 517:523 arg2 glycans 509:515 host glycans 504:515 host glycans present in the glycocalyx of epithelial cells and extracellular mucus 504:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 3 66 attach present 517:523 arg1 glycocalyx 532:541 the glycocalyx 528:541 the glycocalyx of epithelial cells and extracellular mucus 528:585 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 6 67 theme type 1143:1146 arg1 Lewis-a 1167:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 the terminal type 1 glycan structure Lewis-a 1130:1173 Our results demonstrated that the FUT2 determines the O-glycosylation pattern of Muc5ac, with Fut2 knock-out leading to a marked decrease in α1,2-fucosylated structures and increased expression of the terminal type 1 glycan structure Lewis-a. 27161092 1 68 theme mucosal 208:214 arg1 epithelium 216:225 the mucosal epithelium 204:225 the mucosal epithelium 204:225 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 27161092 3 69 theme epithelial 546:555 arg1 cells 557:561 epithelial cells 546:561 epithelial cells 546:561 The carcinogenic bacteria Helicobacter pylori colonize the stomach through binding to host glycans present in the glycocalyx of epithelial cells and extracellular mucus. 27161092 5 70 theme glycoengineered 825:839 arg1 models 846:851 glycoengineered mice models 825:851 glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype 825:930 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 4 71 theme increased 721:729 arg1 risk 731:734 increased risk 721:734 increased risk for overt disease 721:752 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 72 theme mucus 651:655 arg1 layer 657:661 the gastric mucus layer 639:661 the gastric mucus layer 639:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 8 73 theme adhesin 1408:1414 arg1 binding 1382:1388 gastric mucosal binding 1366:1388 gastric mucosal binding 1366:1388 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 8 73 theme adhesin 1408:1414 arg1 feature 1439:1445 a recognized feature 1426:1445 a recognized feature of pathogenicity 1426:1462 Finally, we demonstrated that loss of mucin FUT2-mediated fucosylation impairs gastric mucosal binding of H. pylori BabA adhesin, which is a recognized feature of pathogenicity. 27161092 4 74 theme gastric 643:649 arg1 layer 657:661 the gastric mucus layer 639:661 the gastric mucus layer 639:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 75 theme H. 693:694 arg1 pylori 696:701 H. pylori 693:701 H. pylori 693:701 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 7 76 theme Lewis-a 1253:1259 arg1 expression 1239:1248 the expression 1235:1248 the expression of Lewis-a in murine gastric mucosa 1235:1284 Importantly, for the first time, we structurally validated the expression of Lewis-a in murine gastric mucosa. 27161092 5 77 theme O-glycosylation 786:800 arg1 profile 802:808 the O-glycosylation profile 782:808 the O-glycosylation profile of Muc5ac 782:818 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 5 78 gly O-glycosylation 786:800 arg1 Muc5ac 813:818 Muc5ac 813:818 Muc5ac 813:818 In this study we unraveled the O-glycosylation profile of Muc5ac from glycoengineered mice models lacking the FUT2 enzyme and therefore mimicking a non-secretor human phenotype. 27161092 4 79 theme main 621:624 arg1 mucin 608:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin 588:612 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 4 79 theme main 621:624 arg1 component 626:634 the main component 617:634 the main component of the gastric mucus layer 617:661 The secreted MUC5AC mucin is the main component of the gastric mucus layer, and BabA-mediated binding of H. pylori to MUC5AC confers increased risk for overt disease. 27161092 1 80 theme biochemical 232:242 arg1 aggressions 259:269 biochemical and mechanical aggressions 232:269 biochemical and mechanical aggressions 232:269 The gastrointestinal tract is lined by a thick and complex layer of mucus that protects the mucosal epithelium from biochemical and mechanical aggressions. 28246170 3 0 theme protein 616:622 arg1 PsrP 624:627 a mucin-like serine-rich repeat protein PsrP 584:627 a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 584:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 1 1 theme structural 288:297 arg1 complexity 299:308 their structural complexity 282:308 their structural complexity 282:308 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 6 2 theme full-length 1226:1236 arg1 GlyA 1238:1241 full-length GlyA 1226:1241 full-length GlyA 1226:1241 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 7 3 theme multiple 1370:1377 arg1 sites 1379:1383 multiple sites 1370:1383 multiple sites along the serine-rich repeats 1370:1413 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 3 4 theme serine-rich 597:607 arg1 PsrP 624:627 a mucin-like serine-rich repeat protein PsrP 584:627 a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 584:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 2 5 theme mucin-like 346:355 arg1 proteins 357:364 mucin-like proteins 346:364 mucin-like proteins 346:364 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 1 6 theme specific 226:233 arg1 functions 235:243 the specific functions 222:243 the specific functions of these glycans 222:260 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 8 7 theme UDP-complexed 1500:1512 arg1 GlyE 1514:1517 apo- and UDP-complexed GlyE 1491:1517 apo- and UDP-complexed GlyE combined with structural analyses 1491:1551 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 6 8 theme fourth 1285:1290 arg1 step 1292:1295 only the fourth step 1276:1295 only the fourth step 1276:1295 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 5 9 theme in 900:901 arg1 assays 935:940 in vivo and in vitro glycosylation assays 900:940 in vivo and in vitro glycosylation assays combined with hydrolytic activity assays 900:981 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 5 10 theme glycosylation 1095:1107 arg1 steps 1086:1090 the third and fourth steps 1065:1090 the third and fourth steps of glycosylation 1065:1107 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 3 11 theme Streptococcus 653:665 arg1 pneumoniae 667:676 the human pathogen Streptococcus pneumoniae TIGR4 634:682 the human pathogen Streptococcus pneumoniae TIGR4 634:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 5 12 theme activity 967:974 arg1 assays 976:981 hydrolytic activity assays 956:981 hydrolytic activity assays 956:981 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 9 13 theme complete 1767:1774 arg1 pathway 1790:1796 the complete glycosylation pathway 1763:1796 the complete glycosylation pathway of a bacterial glycoprotein 1763:1824 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 8 14 theme structural 1533:1542 arg1 analyses 1544:1551 structural analyses 1533:1551 structural analyses 1533:1551 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 1 15 theme important 146:154 arg1 modification 175:186 an important post-translational modification 143:186 an important post-translational modification in all organisms 143:203 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 1 15 theme important 146:154 arg1 O-glycosylation 124:138 Protein O-glycosylation 116:138 Protein O-glycosylation 116:138 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 6 16 theme GlyD 1251:1254 arg1 domain 1260:1265 the GlyD GT8 domain 1247:1265 the GlyD GT8 domain 1247:1265 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 8 17 theme novel 1562:1566 arg1 domain 1591:1596 a novel Rossmann-fold "add-on" domain 1560:1596 a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another 1560:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 8 17 theme novel 1562:1566 arg1 module 1643:1648 a universal module 1631:1648 a universal module shared by GlyD, GlyE, and GlyA 1631:1679 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 0 18 theme serine-rich 83:93 arg1 PsrP 110:113 the pneumococcal serine-rich repeat protein PsrP 66:113 the pneumococcal serine-rich repeat protein PsrP 66:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 9 19 theme glycoprotein 1813:1824 arg1 pathway 1790:1796 the complete glycosylation pathway 1763:1796 the complete glycosylation pathway of a bacterial glycoprotein 1763:1824 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 4 20 theme first 822:826 arg1 reactions 832:840 the first two reactions 818:840 the first two reactions to form a unified disaccharide core structure 818:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 1 21 theme Protein 116:122 arg1 modification 175:186 an important post-translational modification 143:186 an important post-translational modification in all organisms 143:203 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 1 21 theme Protein 116:122 arg1 O-glycosylation 124:138 Protein O-glycosylation 116:138 Protein O-glycosylation 116:138 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 0 22 theme protein 102:108 arg1 PsrP 110:113 the pneumococcal serine-rich repeat protein PsrP 66:113 the pneumococcal serine-rich repeat protein PsrP 66:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 8 23 theme " 1589:1589 arg1 domain 1591:1596 a novel Rossmann-fold "add-on" domain 1560:1596 a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another 1560:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 8 23 theme " 1589:1589 arg1 module 1643:1648 a universal module 1631:1648 a universal module shared by GlyD, GlyE, and GlyA 1631:1679 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 5 24 theme core 1047:1050 arg1 structure 1052:1060 this core structure 1042:1060 this core structure 1042:1060 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 6 25 theme GlyE 1140:1143 arg1 proteins 1154:1161 the full-length GlyE and GlyG proteins 1124:1161 the full-length GlyE and GlyG proteins 1124:1161 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 9 26 theme testable 1838:1845 arg1 hypothesis 1847:1856 a testable hypothesis 1836:1856 a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly 1836:1924 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 2 27 theme glycosylation 473:485 arg1 patterns 487:494 the glycosylation patterns 469:494 the glycosylation patterns 469:494 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 7 28 theme serine-rich 1395:1405 arg1 repeats 1407:1413 the serine-rich repeats 1391:1413 the serine-rich repeats 1391:1413 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 2 29 dep challenge 388:396 arg1 modified 410:417 modified 410:417 are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood 406:518 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 4 30 theme core 873:876 arg1 structure 878:886 a unified disaccharide core structure 850:886 a unified disaccharide core structure 850:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 0 31 theme enzymatic 13:21 arg1 pathway 23:29 the enzymatic pathway 9:29 the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP 9:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 5 32 theme capable 1020:1026 arg1 glycosyltransferases 999:1018 the glycosyltransferases 995:1018 the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation 995:1107 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 1 33 theme post-translational 156:173 arg1 modification 175:186 an important post-translational modification 143:186 an important post-translational modification in all organisms 143:203 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 1 33 theme post-translational 156:173 arg1 O-glycosylation 124:138 Protein O-glycosylation 116:138 Protein O-glycosylation 116:138 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 9 34 theme glycosyltransferase 1865:1883 arg1 coordination 1885:1896 glycosyltransferase coordination 1865:1896 glycosyltransferase coordination 1865:1896 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 5 35 dep in 912:913 arg1 vitro 915:919 vitro 915:919 vitro 915:919 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 5 36 theme fourth 1079:1084 arg1 steps 1086:1090 the third and fourth steps 1065:1090 the third and fourth steps of glycosylation 1065:1107 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 7 37 theme core 1352:1355 arg1 structure 1357:1365 the disaccharide core structure 1335:1365 the disaccharide core structure 1335:1365 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 8 38 theme crystal 1469:1475 arg1 structures 1477:1486 crystal structures 1469:1486 crystal structures of apo- and UDP-complexed GlyE combined with structural analyses 1469:1551 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 6 39 theme GlyD 1171:1174 arg1 domain 1184:1189 the GlyD DUF1792 domain 1167:1189 the GlyD DUF1792 domain 1167:1189 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 4 40 theme unified 852:858 arg1 structure 878:886 a unified disaccharide core structure 850:886 a unified disaccharide core structure 850:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 8 41 theme peptide 1696:1702 arg1 acceptor 1704:1711 the peptide acceptor 1692:1711 the peptide acceptor from one enzyme to another 1692:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 5 42 theme third 1069:1073 arg1 steps 1086:1090 the third and fourth steps 1065:1090 the third and fourth steps of glycosylation 1065:1107 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 2 43 gly glycosylation 329:341 arg1 proteins 357:364 mucin-like proteins 346:364 mucin-like proteins 346:364 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 3 44 theme O-glycosylation 557:571 arg1 pathway 573:579 the O-glycosylation pathway 553:579 the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 553:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 6 45 theme GlyG 1149:1152 arg1 proteins 1154:1161 the full-length GlyE and GlyG proteins 1124:1161 the full-length GlyE and GlyG proteins 1124:1161 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 3 46 from pathway 573:579 arg1 pneumoniae 667:676 the human pathogen Streptococcus pneumoniae TIGR4 634:682 the human pathogen Streptococcus pneumoniae TIGR4 634:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 3 47 theme repeat 609:614 arg1 PsrP 624:627 a mucin-like serine-rich repeat protein PsrP 584:627 a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 584:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 3 48 theme mucin-like 586:595 arg1 PsrP 624:627 a mucin-like serine-rich repeat protein PsrP 584:627 a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 584:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 4 49 theme 3 730:730 arg1 function 718:725 the function 714:725 the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure 714:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 2 50 theme proteins 357:364 arg1 glycosylation 329:341 the glycosylation 325:341 the glycosylation of mucin-like proteins 325:364 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 5 51 theme in 912:913 arg1 assays 935:940 in vivo and in vitro glycosylation assays 900:940 in vivo and in vitro glycosylation assays combined with hydrolytic activity assays 900:981 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 3 52 theme PsrP 624:627 arg1 pathway 573:579 the O-glycosylation pathway 553:579 the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 553:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 3 53 theme human 638:642 arg1 pneumoniae 667:676 the human pathogen Streptococcus pneumoniae TIGR4 634:682 the human pathogen Streptococcus pneumoniae TIGR4 634:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 8 54 theme GlyE 1514:1517 arg1 structures 1477:1486 crystal structures 1469:1486 crystal structures of apo- and UDP-complexed GlyE combined with structural analyses 1469:1551 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 3 55 theme pathogen 644:651 arg1 pneumoniae 667:676 the human pathogen Streptococcus pneumoniae TIGR4 634:682 the human pathogen Streptococcus pneumoniae TIGR4 634:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 5 56 theme hydrolytic 956:965 arg1 assays 976:981 hydrolytic activity assays 956:981 hydrolytic activity assays 956:981 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 9 57 theme glycosylation 1776:1788 arg1 pathway 1790:1796 the complete glycosylation pathway 1763:1796 the complete glycosylation pathway of a bacterial glycoprotein 1763:1824 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 6 58 theme GT8 1256:1258 arg1 domain 1260:1265 the GlyD GT8 domain 1247:1265 the GlyD GT8 domain 1247:1265 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 7 59 theme different 1315:1323 arg1 sugars 1325:1330 different sugars 1315:1330 different sugars 1315:1330 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 0 60 theme pneumococcal 70:81 arg1 PsrP 110:113 the pneumococcal serine-rich repeat protein PsrP 66:113 the pneumococcal serine-rich repeat protein PsrP 66:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 9 61 gly glycoprotein 1813:1824 arg1 glycoprotein 1813:1824 a bacterial glycoprotein 1801:1824 a bacterial glycoprotein 1801:1824 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 5 62 theme glycosylation 921:933 arg1 assays 935:940 in vivo and in vitro glycosylation assays 900:940 in vivo and in vitro glycosylation assays combined with hydrolytic activity assays 900:981 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 8 63 theme Rossmann-fold 1568:1580 arg1 domain 1591:1596 a novel Rossmann-fold "add-on" domain 1560:1596 a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another 1560:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 8 63 theme Rossmann-fold 1568:1580 arg1 module 1643:1648 a universal module 1631:1648 a universal module shared by GlyD, GlyE, and GlyA 1631:1679 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 4 64 theme glycosyltransferases 742:761 arg1 glycosyltransferases 742:761 the 10 glycosyltransferases 735:761 the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure 735:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 4 64 theme glycosyltransferases 742:761 arg1 3 730:730 3 730:730 3 730:730 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 0 65 theme repeat 95:100 arg1 PsrP 110:113 the pneumococcal serine-rich repeat protein PsrP 66:113 the pneumococcal serine-rich repeat protein PsrP 66:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 9 66 theme bacterial 1803:1811 arg1 glycoprotein 1813:1824 a bacterial glycoprotein 1801:1824 a bacterial glycoprotein 1801:1824 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 8 67 theme add-on 1583:1588 arg1 domain 1591:1596 a novel Rossmann-fold "add-on" domain 1560:1596 a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another 1560:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 8 67 theme add-on 1583:1588 arg1 module 1643:1648 a universal module 1631:1648 a universal module shared by GlyD, GlyE, and GlyA 1631:1679 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 0 68 theme PsrP 110:113 arg1 O-glycosylation 47:61 polymorphic O-glycosylation 35:61 polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP 35:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 7 69 theme sugars 1325:1330 arg1 Incorporation 1298:1310 Incorporation 1298:1310 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats 1298:1413 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 1 70 from modification 175:186 arg1 organisms 195:203 all organisms 191:203 all organisms 191:203 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 6 71 theme full-length 1128:1138 arg1 proteins 1154:1161 the full-length GlyE and GlyG proteins 1124:1161 the full-length GlyE and GlyG proteins 1124:1161 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 8 72 from enzyme 1722:1727 arg1 acceptor 1704:1711 the peptide acceptor 1692:1711 the peptide acceptor from one enzyme to another 1692:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 2 73 theme numerous 419:426 arg1 times 428:432 numerous times 419:432 numerous times 419:432 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 7 74 theme polymorphic 1435:1445 arg1 product 1447:1453 a highly polymorphic product 1426:1453 a highly polymorphic product 1426:1453 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 3 75 from pneumoniae 667:676 arg1 pathway 573:579 the O-glycosylation pathway 553:579 the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 553:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 3 75 from pneumoniae 667:676 arg1 PsrP 624:627 a mucin-like serine-rich repeat protein PsrP 584:627 a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4 584:682 Here we systematically explored the O-glycosylation pathway of a mucin-like serine-rich repeat protein PsrP from the human pathogen Streptococcus pneumoniae TIGR4. 28246170 0 76 theme polymorphic 35:45 arg1 O-glycosylation 47:61 polymorphic O-glycosylation 35:61 polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP 35:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 7 77 theme disaccharide 1339:1350 arg1 structure 1357:1365 the disaccharide core structure 1335:1365 the disaccharide core structure 1335:1365 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 8 78 theme universal 1633:1641 arg1 domain 1591:1596 a novel Rossmann-fold "add-on" domain 1560:1596 a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another 1560:1738 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 8 78 theme universal 1633:1641 arg1 module 1643:1648 a universal module 1631:1648 a universal module shared by GlyD, GlyE, and GlyA 1631:1679 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 28246170 4 79 theme disaccharide 860:871 arg1 structure 878:886 a unified disaccharide core structure 850:886 a unified disaccharide core structure 850:886 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 9 80 theme glycan 1910:1915 arg1 assembly 1917:1924 glycan assembly 1910:1924 glycan assembly 1910:1924 These findings define the complete glycosylation pathway of a bacterial glycoprotein and offer a testable hypothesis of how glycosyltransferase coordination facilitates glycan assembly. 28246170 2 81 theme particular 377:386 arg1 challenge 388:396 a particular challenge 375:396 a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood 375:518 Understanding the glycosylation of mucin-like proteins presents a particular challenge as they are modified numerous times with both the enzymes involved and the glycosylation patterns being poorly understood. 28246170 6 82 theme DUF1792 1176:1182 arg1 domain 1184:1189 the GlyD DUF1792 domain 1167:1189 the GlyD DUF1792 domain 1167:1189 Specifically, the full-length GlyE and GlyG proteins and the GlyD DUF1792 domain participate in both steps, whereas full-length GlyA and the GlyD GT8 domain catalyze only the fourth step. 28246170 4 83 theme Previous 685:692 arg1 works 694:698 Previous works 685:698 Previous works 685:698 Previous works have assigned the function of 3 of the 10 glycosyltransferases thought to modify PsrP, GtfA/B, and Gtf3 as catalyzing the first two reactions to form a unified disaccharide core structure. 28246170 5 84 dep in 900:901 arg1 vivo 903:906 vivo 903:906 vivo 903:906 We now use in vivo and in vitro glycosylation assays combined with hydrolytic activity assays to identify the glycosyltransferases capable of decorating this core structure in the third and fourth steps of glycosylation. 28246170 1 85 theme glycans 254:260 arg1 functions 235:243 the specific functions 222:243 the specific functions of these glycans 222:260 Protein O-glycosylation is an important post-translational modification in all organisms, but deciphering the specific functions of these glycans is difficult due to their structural complexity. 28246170 0 86 gly O-glycosylation 47:61 arg1 PsrP 110:113 the pneumococcal serine-rich repeat protein PsrP 66:113 the pneumococcal serine-rich repeat protein PsrP 66:113 Defining the enzymatic pathway for polymorphic O-glycosylation of the pneumococcal serine-rich repeat protein PsrP. 28246170 7 87 from sites 1379:1383 arg1 Incorporation 1298:1310 Incorporation 1298:1310 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats 1298:1413 Incorporation of different sugars to the disaccharide core structure at multiple sites along the serine-rich repeats results in a highly polymorphic product. 28246170 8 88 theme apo- 1491:1494 arg1 GlyE 1514:1517 apo- and UDP-complexed GlyE 1491:1517 apo- and UDP-complexed GlyE combined with structural analyses 1491:1551 Furthermore, crystal structures of apo- and UDP-complexed GlyE combined with structural analyses reveal a novel Rossmann-fold "add-on" domain that we speculate to function as a universal module shared by GlyD, GlyE, and GlyA to forward the peptide acceptor from one enzyme to another. 25498018 0 0 theme recombinant 90:100 arg1 CLEC-2 102:107 recombinant CLEC-2 90:107 recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 90:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 1 from differences 15:25 arg1 N-glycosylation 30:44 N-glycosylation 30:44 N-glycosylation 30:44 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 4 2 theme direct 841:846 arg1 elucidation 848:858 sensitive and direct elucidation 827:858 sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2 827:917 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 1 3 theme weight 334:339 arg1 form 347:350 a higher molecular weight (HMW) and a lower molecular weight (LMW) form 280:350 a higher molecular weight (HMW) and a lower molecular weight (LMW) form 280:350 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 7 4 theme samples 1655:1661 arg1 samples 1655:1661 samples 1655:1661 samples 1655:1661 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 7 4 theme samples 1655:1661 arg1 amount 1645:1650 small amount 1639:1650 small amount of samples 1639:1661 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 3 5 theme N-glycosylation 653:667 arg1 difference 669:678 the N-glycosylation difference 649:678 the N-glycosylation difference 649:678 This study aimed to explore an easy method to directly elucidate the N-glycosylation difference by employing a glycoproteomics approach. 25498018 0 6 theme platelet 219:226 arg1 surface 228:234 the platelet surface 215:234 the platelet surface 215:234 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 4 7 theme elucidation 848:858 arg1 coupling 734:741 The off-line coupling 721:741 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer 721:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 4 7 theme elucidation 848:858 arg1 capable 816:822 capable 816:822 capable 816:822 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 5 8 gly glycosylation 1060:1072 arg2 Asn 1080:1082 Asn 134 1080:1086 Asn 134 1080:1086 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 8 gly glycosylation 1060:1072 arg2 site 1074:1077 a specific glycosylation site 1049:1077 a specific glycosylation site 1049:1077 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 9 theme tetra-antennary 1189:1203 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 2 10 theme ligand-binding 449:462 arg1 abilities 464:472 different ligand-binding abilities 439:472 different ligand-binding abilities 439:472 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 0 11 from Elucidation 0:10 arg1 N-glycosylation 30:44 N-glycosylation 30:44 N-glycosylation 30:44 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 12 theme protein 374:380 arg1 core 382:385 the same protein core 365:385 the same protein core 365:385 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 6 13 theme biological 1403:1412 arg1 functions 1414:1422 biological functions 1403:1422 biological functions of CLEC-2 1403:1432 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 4 14 theme oligosaccharide 969:983 arg1 structures 985:994 their oligosaccharide structures 963:994 their oligosaccharide structures 963:994 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 0 15 theme CLEC-2 102:107 arg1 forms 81:85 different molecular weight forms 54:85 different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 54:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 2 16 theme CLEC-2 525:530 arg1 glycosylation 508:520 the differential glycosylation 491:520 the differential glycosylation of CLEC-2 491:530 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 4 17 theme off-line 725:732 arg1 coupling 734:741 The off-line coupling 721:741 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer 721:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 4 17 theme off-line 725:732 arg1 capable 816:822 capable 816:822 capable 816:822 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 5 18 theme N-linked 1206:1213 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 0 19 theme MALDI 115:119 arg1 MS. 128:130 LC MALDI tandem MS. 112:130 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 20 theme lower 318:322 arg1 LMW 342:344 LMW 342:344 LMW 342:344 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 1 20 theme lower 318:322 arg1 weight 334:339 a lower molecular weight 316:339 a lower molecular weight (LMW) 316:345 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 4 21 theme nano-LC 746:752 arg1 coupling 734:741 The off-line coupling 721:741 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer 721:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 4 21 theme nano-LC 746:752 arg1 capable 816:822 capable 816:822 capable 816:822 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 7 22 gly glycoprotein 1617:1628 arg1 glycoprotein 1617:1628 target glycoprotein 1610:1628 target glycoprotein 1610:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 6 23 theme new 1356:1358 arg1 insights 1360:1367 new insights 1356:1367 new insights into the underlying mechanisms of biological functions of CLEC-2 1356:1432 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 0 24 theme LC 112:113 arg1 MS. 128:130 LC MALDI tandem MS. 112:130 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 25 theme molecular 324:332 arg1 LMW 342:344 LMW 342:344 LMW 342:344 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 1 25 theme molecular 324:332 arg1 weight 334:339 a lower molecular weight 316:339 a lower molecular weight (LMW) 316:345 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 0 26 theme identified 182:191 arg1 receptor 193:200 a newly identified receptor 174:200 a newly identified receptor expressed on the platelet surface 174:234 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 26 theme identified 182:191 arg1 Elucidation 0:10 Elucidation 0:10 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 0:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 7 27 from differences 1569:1579 arg1 N-glycosylation 1591:1605 direct N-glycosylation 1584:1605 direct N-glycosylation of target glycoprotein 1584:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 5 28 theme complex 1140:1146 arg1 types 1148:1152 complex types 1140:1152 complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form 1140:1298 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 6 29 theme CLEC-2 1427:1432 arg1 functions 1414:1422 biological functions 1403:1422 biological functions of CLEC-2 1403:1432 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 0 30 from N-glycosylation 30:44 arg1 receptor 193:200 a newly identified receptor 174:200 a newly identified receptor expressed on the platelet surface 174:234 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 30 from N-glycosylation 30:44 arg1 Elucidation 0:10 Elucidation 0:10 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 0:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 4 31 theme sensitive 827:835 arg1 elucidation 848:858 sensitive and direct elucidation 827:858 sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2 827:917 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 7 32 theme glycoprotein 1617:1628 arg1 N-glycosylation 1591:1605 direct N-glycosylation 1584:1605 direct N-glycosylation of target glycoprotein 1584:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 6 33 from difference 1314:1323 arg1 glycosylation 1328:1340 glycosylation 1328:1340 glycosylation 1328:1340 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 6 34 theme underlying 1378:1387 arg1 mechanisms 1389:1398 the underlying mechanisms 1374:1398 the underlying mechanisms of biological functions of CLEC-2 1374:1432 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 5 35 theme glycosylation 1060:1072 arg1 Asn 1080:1082 Asn 134 1080:1086 Asn 134 1080:1086 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 35 theme glycosylation 1060:1072 arg1 site 1074:1077 a specific glycosylation site 1049:1077 a specific glycosylation site 1049:1077 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 2 36 theme distinct 563:570 arg1 glycoforms 572:581 functionally distinct glycoforms 550:581 functionally distinct glycoforms 550:581 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 7 37 theme small 1639:1643 arg1 samples 1655:1661 samples 1655:1661 samples 1655:1661 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 7 37 theme small 1639:1643 arg1 amount 1645:1650 small amount 1639:1650 small amount of samples 1639:1661 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 6 38 theme functions 1414:1422 arg1 mechanisms 1389:1398 the underlying mechanisms 1374:1398 the underlying mechanisms of biological functions of CLEC-2 1374:1432 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 4 39 theme mass 775:778 arg1 spectrometer 780:791 a MALDI-QIT-TOF mass spectrometer 759:791 a MALDI-QIT-TOF mass spectrometer 759:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 0 40 theme MS. 128:130 arg1 receptor 151:158 LC MALDI tandem MS. C-type lectin-like receptor 2 112:160 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 40 theme MS. 128:130 arg1 CLEC-2 163:168 CLEC-2 163:168 CLEC-2 163:168 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 41 theme same 369:372 arg1 core 382:385 the same protein core 365:385 the same protein core 365:385 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 0 42 theme differences 15:25 arg1 receptor 193:200 a newly identified receptor 174:200 a newly identified receptor expressed on the platelet surface 174:234 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 42 theme differences 15:25 arg1 Elucidation 0:10 Elucidation 0:10 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 0:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 3 43 theme glycoproteomics 695:709 arg1 approach 711:718 a glycoproteomics approach 693:718 a glycoproteomics approach 693:718 This study aimed to explore an easy method to directly elucidate the N-glycosylation difference by employing a glycoproteomics approach. 25498018 0 44 theme tandem 121:126 arg1 MS. 128:130 LC MALDI tandem MS. 112:130 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 4 45 theme LMW 908:910 arg1 CLEC-2 912:917 LMW CLEC-2 908:917 LMW CLEC-2 908:917 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 5 46 gly glycosylated 1104:1115 arg2 Asn 1080:1082 Asn 134 1080:1086 Asn 134 1080:1086 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 46 gly glycosylated 1104:1115 arg1 Asn 1080:1082 Asn 134 1080:1086 Asn 134 1080:1086 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 46 gly glycosylated 1104:1115 arg2 site 1074:1077 a specific glycosylation site 1049:1077 a specific glycosylation site 1049:1077 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 46 gly glycosylated 1104:1115 arg1 site 1074:1077 a specific glycosylation site 1049:1077 a specific glycosylation site 1049:1077 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 46 gly glycosylated 1104:1115 arg1 forms 1128:1132 the two forms 1120:1132 the two forms 1120:1132 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 47 theme tri-antennary 1171:1183 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 0 48 theme lectin-like 139:149 arg1 receptor 151:158 LC MALDI tandem MS. C-type lectin-like receptor 2 112:160 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 48 theme lectin-like 139:149 arg1 CLEC-2 163:168 CLEC-2 163:168 CLEC-2 163:168 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 7 49 theme target 1610:1615 arg1 glycoprotein 1617:1628 target glycoprotein 1610:1628 target glycoprotein 1610:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 5 50 theme LMW 1291:1293 arg1 form 1295:1298 the LMW form 1287:1298 the LMW form 1287:1298 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 4 51 theme glycosylation 1004:1016 arg1 sites 1018:1022 the glycosylation sites 1000:1022 the glycosylation sites 1000:1022 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 0 52 theme C-type 132:137 arg1 receptor 151:158 LC MALDI tandem MS. C-type lectin-like receptor 2 112:160 LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 112:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 0 52 theme C-type 132:137 arg1 CLEC-2 163:168 CLEC-2 163:168 CLEC-2 163:168 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 5 53 theme glycans 1228:1234 arg1 types 1148:1152 complex types 1140:1152 complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form 1140:1298 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 7 54 gly N-glycosylation 1591:1605 arg1 glycoprotein 1617:1628 target glycoprotein 1610:1628 target glycoprotein 1610:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 0 55 theme molecular 64:72 arg1 forms 81:85 different molecular weight forms 54:85 different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 54:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 56 theme higher 282:287 arg1 HMW 307:309 HMW 307:309 HMW 307:309 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 1 56 theme higher 282:287 arg1 weight 299:304 a higher molecular weight 280:304 a higher molecular weight (HMW) 280:310 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 4 57 theme glycosylation 867:879 arg1 difference 881:890 the glycosylation difference 863:890 the glycosylation difference between HMW and LMW CLEC-2 863:917 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 5 58 theme bi-antennary 1157:1168 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 2 59 gly glycosylation 508:520 arg1 CLEC-2 525:530 CLEC-2 525:530 CLEC-2 525:530 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 0 60 theme different 54:62 arg1 forms 81:85 different molecular weight forms 54:85 different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 54:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 1 61 theme molecular 289:297 arg1 HMW 307:309 HMW 307:309 HMW 307:309 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 1 61 theme molecular 289:297 arg1 weight 299:304 a higher molecular weight 280:304 a higher molecular weight (HMW) 280:310 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 6 62 theme observed 1305:1312 arg1 difference 1314:1323 The observed difference 1301:1323 The observed difference in glycosylation 1301:1340 The observed difference in glycosylation might provide new insights into the underlying mechanisms of biological functions of CLEC-2. 25498018 1 63 theme weight 299:304 arg1 form 347:350 a higher molecular weight (HMW) and a lower molecular weight (LMW) form 280:350 a higher molecular weight (HMW) and a lower molecular weight (LMW) form 280:350 It has been reported that CLEC-2 exists as a higher molecular weight (HMW) and a lower molecular weight (LMW) form, which share the same protein core but differ in glycans. 25498018 2 64 theme differential 495:506 arg1 glycosylation 508:520 the differential glycosylation 491:520 the differential glycosylation of CLEC-2 491:530 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 5 65 link N-linked 1206:1213 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 66 theme HMW 1267:1269 arg1 form 1271:1274 the HMW form 1263:1274 the HMW form 1263:1274 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 4 67 theme difference 881:890 arg1 elucidation 848:858 sensitive and direct elucidation 827:858 sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2 827:917 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 5 68 theme specific 1051:1058 arg1 Asn 1080:1082 Asn 134 1080:1086 Asn 134 1080:1086 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 68 theme specific 1051:1058 arg1 site 1074:1077 a specific glycosylation site 1049:1077 a specific glycosylation site 1049:1077 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 69 gly fucosylated 1216:1226 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 5 70 theme fucosylated 1216:1226 arg1 glycans 1228:1234 bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans 1157:1234 glycans 1228:1234 The results reveal that a specific glycosylation site, Asn 134, is differently glycosylated in the two forms, with complex types of bi-antennary, tri-antennary and tetra-antennary, N-linked, fucosylated glycans identified at this site in the HMW form but not in the LMW form. 25498018 3 71 theme easy 615:618 arg1 method 620:625 an easy method 612:625 an easy method to directly elucidate the N-glycosylation difference by employing a glycoproteomics approach 612:718 This study aimed to explore an easy method to directly elucidate the N-glycosylation difference by employing a glycoproteomics approach. 25498018 4 72 gly glycosylation 1004:1016 arg2 sites 1018:1022 the glycosylation sites 1000:1022 the glycosylation sites 1000:1022 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 2 73 contain have 434:437 arg1 forms 418:422 The two forms 410:422 The two forms 410:422 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 2 73 contain have 434:437 arg2 abilities 464:472 different ligand-binding abilities 439:472 different ligand-binding abilities 439:472 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 25498018 4 74 with coupling 734:741 arg1 spectrometer 780:791 a MALDI-QIT-TOF mass spectrometer 759:791 a MALDI-QIT-TOF mass spectrometer 759:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 0 75 theme weight 74:79 arg1 forms 81:85 different molecular weight forms 54:85 different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) 54:169 Elucidation of differences in N-glycosylation between different molecular weight forms of recombinant CLEC-2 by LC MALDI tandem MS. C-type lectin-like receptor 2 (CLEC-2) is a newly identified receptor expressed on the platelet surface. 25498018 4 76 theme MALDI-QIT-TOF 761:773 arg1 spectrometer 780:791 a MALDI-QIT-TOF mass spectrometer 759:791 a MALDI-QIT-TOF mass spectrometer 759:791 The off-line coupling of nano-LC with a MALDI-QIT-TOF mass spectrometer was demonstrated to be capable of sensitive and direct elucidation of the glycosylation difference between HMW and LMW CLEC-2, simultaneously providing information about their oligosaccharide structures and the glycosylation sites. 25498018 7 77 theme direct 1584:1589 arg1 N-glycosylation 1591:1605 direct N-glycosylation 1584:1605 direct N-glycosylation of target glycoprotein 1584:1628 Because of its simplicity and sensitivity, the method explored in this work suggests that it holds promise as a method of elucidating differences in direct N-glycosylation of target glycoprotein, even in small amount of samples. 25498018 2 78 theme different 439:447 arg1 abilities 464:472 different ligand-binding abilities 439:472 different ligand-binding abilities 439:472 The two forms appear to have different ligand-binding abilities, indicating that the differential glycosylation of CLEC-2 possibly produces functionally distinct glycoforms. 26993603 5 0 theme hepatocellular 785:798 arg1 cells 810:814 hepatocellular carcinoma cells 785:814 hepatocellular carcinoma cells 785:814 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 2 1 from impact 489:494 arg1 glycoproteins 517:529 target glycoproteins 510:529 target glycoproteins 510:529 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 1 2 theme disease 141:147 arg1 Protein 102:108 Golgi Protein 73 96:111 Golgi Protein 73 (GP73) 96:118 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 1 2 theme disease 141:147 arg1 glycobiomarker 149:162 a potential liver disease glycobiomarker 123:162 a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function 123:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 5 3 dep motility 756:763 arg1 the 752:754 the 752:754 the 752:754 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 2 4 gly glycosylation 303:315 arg2 sites 317:321 glycosylation sites 303:321 glycosylation sites 303:321 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 2 5 theme high-throughput 349:363 arg1 microarray 372:381 high-throughput lectin microarray 349:381 high-throughput lectin microarray 349:381 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 6 from Asn144 736:741 arg1 Removal 712:718 Removal 712:718 Removal of N-glycans at Asn144 712:741 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 3 7 theme GP73 560:563 arg1 Asn398 607:612 Asn398 607:612 Asn398 607:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 7 theme GP73 560:563 arg1 Asn144 596:601 Asn144 596:601 Asn144 596:601 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 7 theme GP73 560:563 arg1 Asn109 588:593 Asn109 588:593 Asn109 588:593 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 7 theme GP73 560:563 arg1 sites 581:585 three GP73 N-glycosylation sites 554:585 three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398 554:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 6 8 theme Asn144-deleted 1023:1036 arg1 GP73 1038:1041 Asn144-deleted GP73 1023:1041 Asn144-deleted GP73 1023:1041 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 3 9 theme N-glycosylation 565:579 arg1 Asn398 607:612 Asn398 607:612 Asn398 607:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 9 theme N-glycosylation 565:579 arg1 Asn144 596:601 Asn144 596:601 Asn144 596:601 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 9 theme N-glycosylation 565:579 arg1 Asn109 588:593 Asn109 588:593 Asn109 588:593 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 9 theme N-glycosylation 565:579 arg1 sites 581:585 three GP73 N-glycosylation sites 554:585 three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398 554:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 2 10 theme lectin 365:370 arg1 microarray 372:381 high-throughput lectin microarray 349:381 high-throughput lectin microarray 349:381 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 11 theme carcinoma 800:808 arg1 cells 810:814 hepatocellular carcinoma cells 785:814 hepatocellular carcinoma cells 785:814 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 3 12 gly N-glycosylation 565:579 arg2 Asn144 596:601 Asn144 596:601 Asn144 596:601 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 12 gly N-glycosylation 565:579 arg2 Asn109 588:593 Asn109 588:593 Asn109 588:593 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 12 gly N-glycosylation 565:579 arg2 sites 581:585 three GP73 N-glycosylation sites 554:585 three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398 554:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 12 gly N-glycosylation 565:579 arg2 three 554:558 three 554:558 three 554:558 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 12 gly N-glycosylation 565:579 arg2 Asn398 607:612 Asn398 607:612 Asn398 607:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 5 13 theme related 861:867 arg1 inhibition 833:842 inhibition 833:842 inhibition of cell adhesion related to the changes of cell membrane glycosylation 833:913 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 2 14 theme glycoconjugates 427:441 arg1 profiling 414:422 rapid and sensitive profiling 394:422 rapid and sensitive profiling of glycoconjugates 394:441 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 2 15 theme mass 273:276 arg1 spectrometry 278:289 mass spectrometry 273:289 mass spectrometry 273:289 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 16 theme cell 887:890 arg1 glycosylation 901:913 cell membrane glycosylation 887:913 cell membrane glycosylation 887:913 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 2 17 theme sensitive 404:412 arg1 profiling 414:422 rapid and sensitive profiling 394:422 rapid and sensitive profiling of glycoconjugates 394:441 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 18 theme cells 810:814 arg1 motility 756:763 motility 756:763 motility 756:763 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 5 18 theme cells 810:814 arg1 invasiveness 769:780 invasiveness 769:780 invasiveness 769:780 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 1 19 theme comprehensive 175:187 arg1 analyses 189:196 comprehensive analyses 175:196 comprehensive analyses of its glycan structure and glycosylation function 175:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 2 20 theme glycans 499:505 arg1 impact 489:494 the impact 485:494 the impact of glycans on target glycoproteins 485:529 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 1 21 theme Golgi 96:100 arg1 Protein 102:108 Golgi Protein 73 96:111 Golgi Protein 73 (GP73) 96:118 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 1 21 theme Golgi 96:100 arg1 GP73 114:117 GP73 114:117 GP73 114:117 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 1 21 theme Golgi 96:100 arg1 glycobiomarker 149:162 a potential liver disease glycobiomarker 123:162 a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function 123:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 2 22 theme rapid 394:398 arg1 profiling 414:422 rapid and sensitive profiling 394:422 rapid and sensitive profiling of glycoconjugates 394:441 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 6 23 dep progression 1061:1071 arg1 the 1057:1059 the 1057:1059 the 1057:1059 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 2 24 gly glycoproteins 517:529 arg1 glycoproteins 517:529 target glycoproteins 510:529 target glycoproteins 510:529 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 25 theme membrane 892:899 arg1 glycosylation 901:913 cell membrane glycosylation 887:913 cell membrane glycosylation 887:913 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 2 26 theme site-directed 448:460 arg1 mutagenesis 462:472 site-directed mutagenesis 448:472 site-directed mutagenesis 448:472 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 4 27 gly glycoforms 629:638 arg2 Asn144 643:648 Asn144 643:648 Asn144 643:648 We found five glycoforms on Asn144, including biantennary, triantennary and fucosylated glycans. 26993603 2 28 theme glycosylation 303:315 arg1 sites 317:321 glycosylation sites 303:321 glycosylation sites 303:321 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 5 29 theme adhesion 852:859 arg1 inhibition 833:842 inhibition 833:842 inhibition of cell adhesion related to the changes of cell membrane glycosylation 833:913 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 5 30 theme cell 847:850 arg1 adhesion 852:859 cell adhesion 847:859 cell adhesion 847:859 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 6 31 theme glycosylation 991:1003 arg1 relevance 973:981 the functional relevance 958:981 the functional relevance of GP73 glycosylation 958:1003 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 5 32 theme N-glycans 723:731 arg1 Removal 712:718 Removal 712:718 Removal of N-glycans at Asn144 712:741 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 5 33 attach Removal 712:718 arg3 Asn144 736:741 Asn144 736:741 Asn144 736:741 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 5 33 attach Removal 712:718 arg2 N-glycans 723:731 N-glycans 723:731 N-glycans 723:731 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 1 34 theme glycan 205:210 arg1 structure 212:220 glycan structure 205:220 glycan structure 205:220 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 0 35 theme hepatocellular 39:52 arg1 motility 69:76 hepatocellular carcinoma cell motility 39:76 hepatocellular carcinoma cell motility 39:76 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 6 36 theme GP73 986:989 arg1 glycosylation 991:1003 GP73 glycosylation 986:1003 GP73 glycosylation 986:1003 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 1 37 theme structure 212:220 arg1 analyses 189:196 comprehensive analyses 175:196 comprehensive analyses of its glycan structure and glycosylation function 175:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 0 38 gly N-glycosylation 5:19 arg1 Asn144 24:29 Asn144 24:29 Asn144 24:29 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 0 38 gly N-glycosylation 5:19 arg2 Asn144 24:29 Asn144 24:29 Asn144 24:29 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 6 39 theme hepatocellular 1091:1104 arg1 carcinoma 1106:1114 hepatocellular carcinoma 1091:1114 hepatocellular carcinoma 1091:1114 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 0 40 theme cell 64:67 arg1 motility 69:76 hepatocellular carcinoma cell motility 39:76 hepatocellular carcinoma cell motility 39:76 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 5 41 theme glycosylation 901:913 arg1 changes 876:882 the changes 872:882 the changes of cell membrane glycosylation 872:913 Removal of N-glycans at Asn144 enhanced the motility and invasiveness of hepatocellular carcinoma cells, possibly due to inhibition of cell adhesion related to the changes of cell membrane glycosylation. 26993603 4 42 gly fucosylated 691:701 arg1 glycans 703:709 fucosylated glycans 691:709 fucosylated glycans 691:709 We found five glycoforms on Asn144, including biantennary, triantennary and fucosylated glycans. 26993603 0 43 theme carcinoma 54:62 arg1 motility 69:76 hepatocellular carcinoma cell motility 39:76 hepatocellular carcinoma cell motility 39:76 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 6 44 theme functional 962:971 arg1 relevance 973:981 the functional relevance 958:981 the functional relevance of GP73 glycosylation 958:1003 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 2 45 theme glycan 331:336 arg1 structure 338:346 the glycan structure 327:346 the glycan structure 327:346 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 1 46 theme glycosylation 226:238 arg1 function 240:247 glycosylation function 226:247 glycosylation function 226:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 6 47 theme carcinoma 1106:1114 arg1 metastasis 1077:1086 metastasis 1077:1086 metastasis 1077:1086 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 6 47 theme carcinoma 1106:1114 arg1 progression 1061:1071 progression 1061:1071 progression 1061:1071 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 26993603 4 48 theme fucosylated 691:701 arg1 glycans 703:709 fucosylated glycans 691:709 fucosylated glycans 691:709 We found five glycoforms on Asn144, including biantennary, triantennary and fucosylated glycans. 26993603 1 49 theme function 240:247 arg1 analyses 189:196 comprehensive analyses 175:196 comprehensive analyses of its glycan structure and glycosylation function 175:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 4 50 from glycoforms 629:638 arg1 Asn144 643:648 Asn144 643:648 Asn144 643:648 We found five glycoforms on Asn144, including biantennary, triantennary and fucosylated glycans. 26993603 3 51 dep sites 581:585 arg1 Asn398 607:612 Asn398 607:612 Asn398 607:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 51 dep sites 581:585 arg1 Asn144 596:601 Asn144 596:601 Asn144 596:601 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 51 dep sites 581:585 arg1 Asn109 588:593 Asn109 588:593 Asn109 588:593 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 3 51 dep sites 581:585 arg1 sites 581:585 three GP73 N-glycosylation sites 554:585 three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398 554:612 We identified three GP73 N-glycosylation sites: Asn109, Asn144 and Asn398. 26993603 1 52 theme potential 125:133 arg1 Protein 102:108 Golgi Protein 73 96:111 Golgi Protein 73 (GP73) 96:118 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 1 52 theme potential 125:133 arg1 glycobiomarker 149:162 a potential liver disease glycobiomarker 123:162 a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function 123:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 2 53 theme target 510:515 arg1 glycoproteins 517:529 target glycoproteins 510:529 target glycoproteins 510:529 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 0 54 from Asn144 24:29 arg1 N-glycosylation 5:19 GP73 N-glycosylation 0:19 GP73 N-glycosylation at Asn144 0:29 GP73 N-glycosylation at Asn144 reduces hepatocellular carcinoma cell motility and invasiveness. 26993603 1 55 theme liver 135:139 arg1 Protein 102:108 Golgi Protein 73 96:111 Golgi Protein 73 (GP73) 96:118 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 1 55 theme liver 135:139 arg1 glycobiomarker 149:162 a potential liver disease glycobiomarker 123:162 a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function 123:247 Golgi Protein 73 (GP73) is a potential liver disease glycobiomarker warranting comprehensive analyses of its glycan structure and glycosylation function. 26993603 2 56 used used 268:271 arg2 we 265:266 we 265:266 we 265:266 In this study, we used mass spectrometry to identify glycosylation sites and the glycan structure, high-throughput lectin microarray to provide rapid and sensitive profiling of glycoconjugates, and site-directed mutagenesis to clarify the impact of glycans on target glycoproteins in vivo. 26993603 6 57 theme relevance 973:981 arg1 understanding 941:953 our understanding 937:953 our understanding of the functional relevance of GP73 glycosylation 937:1003 This study increases our understanding of the functional relevance of GP73 glycosylation and suggests that Asn144-deleted GP73 can influence the progression and metastasis of hepatocellular carcinoma. 24475074 3 0 gly glycoproteins 657:669 arg1 glycoproteins 657:669 glycoproteins 657:669 glycoproteins 657:669 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 1 theme ≠ 683:683 arg1 NXS/T 675:679 NXS/T 675:679 NXS/T(X ≠ P) 675:686 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 1 theme ≠ 683:683 arg1 P 685:685 X ≠ P 681:685 X ≠ P 681:685 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 8 2 theme many 1646:1649 arg1 PTMs 1651:1654 many PTMs 1646:1654 many PTMs 1646:1654 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 6 3 theme cytosolic 1125:1133 arg1 proteins 1135:1142 secretory and cytosolic proteins 1111:1142 secretory and cytosolic proteins 1111:1142 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 3 4 theme site 566:569 arg1 multiplicity 571:582 site multiplicity 566:582 site multiplicity (backup and status quo) 566:606 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 6 5 from NXS/T 1218:1222 arg1 one-mutation-away 1195:1211 one-mutation-away 1195:1211 one-mutation-away 1195:1211 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 9 6 from proteins 1735:1742 arg1 examination 1700:1710 An examination 1697:1710 An examination of acetyl-Lys sites in proteins 1697:1742 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 11 7 theme A+T 2086:2088 arg1 shift 2096:2100 the A+T → G+C shift 2082:2100 the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation 2082:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 6 8 theme secretory 1111:1119 arg1 proteins 1135:1142 secretory and cytosolic proteins 1111:1142 secretory and cytosolic proteins 1111:1142 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 9 9 theme portions 1841:1848 arg1 asymmetry 1801:1809 asymmetry 1801:1809 asymmetry of the target and recognition portions of modified sites 1801:1866 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 11 10 theme genome-wide 2105:2115 arg1 composition 2128:2138 genome-wide nucleotide composition 2105:2138 genome-wide nucleotide composition during metazoan radiation 2105:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 5 11 from positions 924:932 arg1 sites 909:913 sites 909:913 sites at novel positions 909:932 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 1 12 theme glycoprotein 168:179 arg1 folding 181:187 glycoprotein folding 168:187 glycoprotein folding 168:187 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 7 13 theme N-glycosylation 1426:1440 arg1 number 1447:1452 N-glycosylation site number 1426:1452 N-glycosylation site number 1426:1452 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 9 14 theme modified 1853:1860 arg1 sites 1862:1866 modified sites 1853:1866 modified sites 1853:1866 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 6 15 theme one-mutation-away 1195:1211 arg1 sequences 1185:1193 sequences 1185:1193 sequences one-mutation-away from NXS/T 1185:1222 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 4 16 theme existing 872:879 arg1 Asn 881:883 an existing Asn 869:883 an existing Asn 869:883 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 6 17 theme selective 1262:1270 arg1 pressures 1272:1280 strong selective pressures 1255:1280 strong selective pressures 1255:1280 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 11 18 theme G+C 2092:2094 arg1 shift 2096:2100 the A+T → G+C shift 2082:2100 the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation 2082:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 2 19 gly glycoproteins 216:228 arg1 glycoproteins 216:228 glycoproteins 216:228 glycoproteins 216:228 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 5 20 theme lost 980:983 arg1 sites 985:989 recently lost sites 971:989 recently lost sites 971:989 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 11 21 theme Strong 2023:2028 arg1 pressures 2040:2048 Strong selective pressures 2023:2048 Strong selective pressures on PTMs 2023:2056 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 10 22 theme evolvability 1940:1951 arg1 asymmetry 1903:1911 encoding asymmetry 1894:1911 encoding asymmetry 1894:1911 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 10 22 theme evolvability 1940:1951 arg1 mechanism 1927:1935 an ancient mechanism 1916:1935 an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions 1916:2020 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 6 23 theme vertebrate 1334:1343 arg1 evolution 1345:1353 vertebrate evolution 1334:1353 vertebrate evolution 1334:1353 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 5 24 theme natural 1061:1067 arg1 selection 1069:1077 natural selection 1061:1077 natural selection 1061:1077 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 2 25 with rebuilt 292:298 arg1 sequences 305:313 sequences 305:313 sequences that bind lectins 305:331 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 6 26 theme positions 1317:1325 arg1 pressures 1272:1280 strong selective pressures 1255:1280 strong selective pressures 1255:1280 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 6 26 theme positions 1317:1325 arg1 exploration 1286:1296 exploration 1286:1296 exploration of N-glycosylation positions 1286:1325 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 5 27 theme sequence 1024:1031 arg1 space 1033:1037 sequence space 1024:1037 sequence space 1024:1037 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 5 28 theme novel 918:922 arg1 positions 924:932 novel positions 918:932 novel positions 918:932 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 7 29 gly N-glycosylation 1426:1440 arg2 number 1447:1452 N-glycosylation site number 1426:1452 N-glycosylation site number 1426:1452 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 7 29 gly N-glycosylation 1426:1440 arg2 site 1442:1445 N-glycosylation site number 1426:1452 N-glycosylation site number 1426:1452 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 9 30 theme acetyl-Lys 1715:1724 arg1 sites 1726:1730 acetyl-Lys sites 1715:1730 acetyl-Lys sites in proteins 1715:1742 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 0 31 theme Encoding 0:7 arg1 asymmetry 9:17 Encoding asymmetry 0:17 Encoding asymmetry of the N-glycosylation motif 0:46 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 2 32 theme cell 449:452 arg1 surface 454:460 the cell surface 445:460 the cell surface 445:460 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 9 33 from examination 1700:1710 arg1 proteins 1735:1742 proteins 1735:1742 proteins 1735:1742 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 3 34 theme glycoproteins 657:669 arg1 NXS/T 675:679 NXS/T 675:679 NXS/T(X ≠ P) 675:686 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 34 theme glycoproteins 657:669 arg1 examination 642:652 a phylogenic examination 629:652 a phylogenic examination of glycoproteins 629:669 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 34 theme glycoproteins 657:669 arg1 P 685:685 X ≠ P 681:685 X ≠ P 681:685 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 2 35 theme glycoprotein-lectin 410:428 arg1 interaction 430:440 glycoprotein-lectin interaction 410:440 glycoprotein-lectin interaction 410:440 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 0 36 theme N-glycosylation 26:40 arg1 motif 42:46 the N-glycosylation motif 22:46 the N-glycosylation motif 22:46 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 1 37 located found 111:115 arg1 life 135:138 life 135:138 life 135:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 37 located found 111:115 arg1 domains 124:130 all domains 120:130 all domains of life 120:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 37 located found 111:115 arg2 N-glycosylation 92:106 Protein N-glycosylation 84:106 Protein N-glycosylation 84:106 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 11 38 theme metazoan 2147:2154 arg1 radiation 2156:2164 metazoan radiation 2147:2164 metazoan radiation 2147:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 7 39 theme underlying 1514:1523 arg1 protein 1525:1531 the N-glycans and underlying protein 1496:1531 the N-glycans and underlying protein 1496:1531 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 11 40 from pressures 2040:2048 arg1 PTMs 2053:2056 PTMs 2053:2056 PTMs 2053:2056 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 9 41 theme evolutionary 1762:1773 arg1 dynamics 1775:1782 similar evolutionary dynamics 1754:1782 similar evolutionary dynamics 1754:1782 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 8 42 theme genetic 1560:1566 arg1 topology 1544:1551 the topology 1540:1551 the topology of the genetic code 1540:1571 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 7 43 theme N-glycans 1500:1508 arg1 protein 1525:1531 the N-glycans and underlying protein 1496:1531 the N-glycans and underlying protein 1496:1531 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 3 44 theme X 681:681 arg1 NXS/T 675:679 NXS/T 675:679 NXS/T(X ≠ P) 675:686 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 44 theme X 681:681 arg1 P 685:685 X ≠ P 681:685 X ≠ P 681:685 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 10 45 theme site 2007:2010 arg1 positions 2012:2020 PTM site positions 2003:2020 PTM site positions 2003:2020 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 11 46 theme nucleotide 2117:2126 arg1 composition 2128:2138 genome-wide nucleotide composition 2105:2138 genome-wide nucleotide composition during metazoan radiation 2105:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 7 47 theme adaptive 1466:1473 arg1 interactions 1475:1486 adaptive interactions 1466:1486 adaptive interactions between the N-glycans and underlying protein 1466:1531 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 0 48 gly glycoprotein 60:71 arg1 glycoprotein 60:71 glycoprotein evolution 60:81 glycoprotein evolution 60:81 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 8 49 theme A 1587:1587 arg1 mutation 1574:1581 mutation 1574:1581 mutation of (A) 1574:1588 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 3 50 theme N-glycosylation 688:702 arg1 sites 704:708 N-glycosylation sites 688:708 N-glycosylation sites 688:708 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 1 51 gly glycoprotein 168:179 arg1 glycoprotein 168:179 glycoprotein folding 168:187 glycoprotein folding 168:187 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 9 52 from sites 1726:1730 arg1 proteins 1735:1742 proteins 1735:1742 proteins 1735:1742 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 9 53 with consistent 1785:1794 arg1 asymmetry 1801:1809 asymmetry 1801:1809 asymmetry of the target and recognition portions of modified sites 1801:1866 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 9 54 theme target 1818:1823 arg1 portions 1841:1848 the target and recognition portions 1814:1848 portions 1841:1848 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 7 55 theme site 1442:1445 arg1 number 1447:1452 N-glycosylation site number 1426:1452 N-glycosylation site number 1426:1452 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 7 56 theme proportional 1410:1421 arg1 rates 1404:1408 rates 1404:1408 rates proportional to N-glycosylation site number 1404:1452 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 9 57 theme recognition 1829:1839 arg1 portions 1841:1848 the target and recognition portions 1814:1848 portions 1841:1848 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 11 58 from shift 2096:2100 arg1 composition 2128:2138 genome-wide nucleotide composition 2105:2138 genome-wide nucleotide composition during metazoan radiation 2105:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 8 59 theme PTMs 1651:1654 arg1 Lys 1623:1625 Lys 1623:1625 Lys 1623:1625 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 8 59 theme PTMs 1651:1654 arg1 target 1636:1641 another target 1628:1641 another target of many PTMs 1628:1654 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 11 60 theme → 2090:2090 arg1 shift 2096:2100 the A+T → G+C shift 2082:2100 the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation 2082:2164 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 1 61 contain has 144:146 arg2 role 160:163 a conserved role 148:163 a conserved role 148:163 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 61 contain has 144:146 arg1 N-glycosylation 92:106 Protein N-glycosylation 84:106 Protein N-glycosylation 84:106 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 6 62 dep content 1235:1241 arg1 A 1232:1232 A 1232:1232 A 1232:1232 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 4 63 theme site 803:806 arg1 gain 808:811 site gain 803:811 site gain 803:811 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 6 64 theme intra-species 1083:1095 arg1 comparison 1097:1106 An intra-species comparison 1080:1106 An intra-species comparison of secretory and cytosolic proteins 1080:1142 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 9 65 theme sites 1862:1866 arg1 portions 1841:1848 the target and recognition portions 1814:1848 portions 1841:1848 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 10 66 theme encoding 1894:1901 arg1 asymmetry 1903:1911 encoding asymmetry 1894:1911 encoding asymmetry 1894:1911 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 10 66 theme encoding 1894:1901 arg1 mechanism 1927:1935 an ancient mechanism 1916:1935 an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions 1916:2020 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 7 67 theme secretory 1369:1377 arg1 proteins 1379:1386 secretory proteins 1369:1386 secretory proteins 1369:1386 Furthermore, secretory proteins have evolved at rates proportional to N-glycosylation site number, indicating adaptive interactions between the N-glycans and underlying protein. 24475074 5 68 theme more 1006:1009 arg1 paths 1011:1015 therefore more paths 996:1015 therefore more paths though sequence space 996:1037 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 10 69 theme ancient 1919:1925 arg1 asymmetry 1903:1911 encoding asymmetry 1894:1911 encoding asymmetry 1894:1911 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 10 69 theme ancient 1919:1925 arg1 mechanism 1927:1935 an ancient mechanism 1916:1935 an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions 1916:2020 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 6 70 theme strong 1255:1260 arg1 pressures 1272:1280 strong selective pressures 1255:1280 strong selective pressures 1255:1280 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 8 71 dep genetic 1560:1566 arg1 code 1568:1571 code 1568:1571 code 1568:1571 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 3 72 theme status 596:601 arg1 quo 603:605 status quo 596:605 status quo 596:605 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 2 73 dep rebuilt 292:298 arg1 innovation 337:346 an innovation 334:346 an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface 334:460 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 6 74 from equilibrium 1170:1180 arg1 departure 1155:1163 a departure 1153:1163 a departure from equilibrium 1153:1180 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 5 75 theme sites 985:989 arg1 reversal 959:966 the reversal 955:966 the reversal of recently lost sites 955:989 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 5 75 theme sites 985:989 arg1 paths 1011:1015 therefore more paths 996:1015 therefore more paths though sequence space 996:1037 Thus mutations produce sites at novel positions more frequently than the reversal of recently lost sites, and therefore more paths though sequence space are made available to natural selection. 24475074 8 76 theme -rich 1683:1687 arg1 A 1681:1681 A 1681:1681 A 1681:1681 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 8 76 theme -rich 1683:1687 arg1 codons 1689:1694 two (A)-rich codons 1676:1694 two (A)-rich codons 1676:1694 Given the topology of the genetic code, mutation of (A) is more often nonsynonomous, and Lys, another target of many PTMs, is also encoded by two (A)-rich codons. 24475074 1 77 theme life 135:138 arg1 life 135:138 life 135:138 life 135:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 77 theme life 135:138 arg1 domains 124:130 all domains 120:130 all domains of life 120:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 11 78 theme selective 2030:2038 arg1 pressures 2040:2048 Strong selective pressures 2023:2048 Strong selective pressures on PTMs 2023:2056 Strong selective pressures on PTMs may have contributed to the A+T → G+C shift in genome-wide nucleotide composition during metazoan radiation. 24475074 4 79 theme Site 711:714 arg1 loss 716:719 Site loss 711:719 Site loss 711:719 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 3 80 dep multiplicity 571:582 arg1 backup 585:590 backup 585:590 backup 585:590 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 80 dep multiplicity 571:582 arg1 quo 603:605 status quo 596:605 status quo 596:605 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 6 81 theme proteins 1135:1142 arg1 comparison 1097:1106 An intra-species comparison 1080:1106 An intra-species comparison of secretory and cytosolic proteins 1080:1142 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 1 82 gly N-glycosylation 92:106 arg1 life 135:138 life 135:138 life 135:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 82 gly N-glycosylation 92:106 arg1 domains 124:130 all domains 120:130 all domains of life 120:138 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 1 83 theme conserved 150:158 arg1 role 160:163 a conserved role 148:163 a conserved role 148:163 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 4 84 from Asn 751:753 arg1 mutation 739:746 mutation 739:746 mutation at Asn encoded by two adenosine (A)-rich codons 739:794 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 4 85 theme Asn 881:883 arg1 downstream 855:864 downstream 855:864 downstream 855:864 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 6 86 theme N-glycosylation 1301:1315 arg1 positions 1317:1325 N-glycosylation positions 1301:1325 N-glycosylation positions 1301:1325 An intra-species comparison of secretory and cytosolic proteins revealed a departure from equilibrium in sequences one-mutation-away from NXS/T and in (A) content, indicating strong selective pressures and exploration of N-glycosylation positions during vertebrate evolution. 24475074 1 87 theme Protein 84:90 arg1 N-glycosylation 92:106 Protein N-glycosylation 84:106 Protein N-glycosylation 84:106 Protein N-glycosylation is found in all domains of life and has a conserved role in glycoprotein folding and stability. 24475074 9 88 theme sites 1726:1730 arg1 examination 1700:1710 An examination 1697:1710 An examination of acetyl-Lys sites in proteins 1697:1742 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 2 89 theme structural 371:380 arg1 diversity 382:390 structural diversity 371:390 structural diversity 371:390 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 2 90 theme interaction 430:440 arg1 redundancy 396:405 redundancy 396:405 redundancy 396:405 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 2 90 theme interaction 430:440 arg1 diversity 382:390 structural diversity 371:390 structural diversity 371:390 In animals, glycoproteins transit through the Golgi where the N-glycans are trimmed and rebuilt with sequences that bind lectins, an innovation that greatly increases structural diversity and redundancy of glycoprotein-lectin interaction at the cell surface. 24475074 0 91 theme motif 42:46 arg1 asymmetry 9:17 Encoding asymmetry 0:17 Encoding asymmetry of the N-glycosylation motif 0:46 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 9 92 theme similar 1754:1760 arg1 dynamics 1775:1782 similar evolutionary dynamics 1754:1782 similar evolutionary dynamics 1754:1782 An examination of acetyl-Lys sites in proteins indicated similar evolutionary dynamics, consistent with asymmetry of the target and recognition portions of modified sites. 24475074 3 93 gly N-glycosylation 688:702 arg2 sites 704:708 N-glycosylation sites 688:708 N-glycosylation sites 688:708 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 94 theme phylogenic 631:640 arg1 examination 642:652 a phylogenic examination 629:652 a phylogenic examination of glycoproteins 629:669 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 0 95 theme glycoprotein 60:71 arg1 evolution 73:81 glycoprotein evolution 60:81 glycoprotein evolution 60:81 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 4 96 theme -rich 783:787 arg1 codons 789:794 two adenosine (A)-rich codons 766:794 two adenosine (A)-rich codons 766:794 Site loss is more likely by mutation at Asn encoded by two adenosine (A)-rich codons, while site gain is more probable by generating Ser or Thr downstream of an existing Asn. 24475074 0 97 gly N-glycosylation 26:40 arg2 motif 42:46 the N-glycosylation motif 22:46 the N-glycosylation motif 22:46 Encoding asymmetry of the N-glycosylation motif facilitates glycoprotein evolution. 24475074 3 98 theme natural 487:493 arg1 tension 495:501 the natural tension 483:501 the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) 483:606 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 99 dep examination 642:652 arg1 sites 704:708 N-glycosylation sites 688:708 N-glycosylation sites 688:708 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 10 100 theme PTM 2003:2005 arg1 positions 2012:2020 PTM site positions 2003:2020 PTM site positions 2003:2020 Our results suggest that encoding asymmetry is an ancient mechanism of evolvability that increases diversity and experimentation with PTM site positions. 24475074 3 101 theme glycan-protein 533:546 arg1 diversity 522:530 diversity 522:530 diversity (glycan-protein interactions) 522:560 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 24475074 3 101 theme glycan-protein 533:546 arg1 interactions 548:559 glycan-protein interactions 533:559 glycan-protein interactions 533:559 Here we ask whether the natural tension between increasing diversity (glycan-protein interactions) and site multiplicity (backup and status quo) might be revealed by a phylogenic examination of glycoproteins and NXS/T(X ≠ P) N-glycosylation sites. 29212317 0 0 theme Density 75:81 arg1 Lipoprotein 83:93 High Density Lipoprotein 70:93 High Density Lipoprotein Particles 70:103 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 4 1 theme protein 700:706 arg1 factor 610:615 A key factor 604:615 A key factor to consider when studying the structure and composition of plasma particles 604:691 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 4 1 theme protein 700:706 arg1 glycosylation 708:720 the protein glycosylation 696:720 the protein glycosylation 696:720 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 3 2 theme particle 492:499 arg1 composition 501:511 HDL particle composition 488:511 HDL particle composition 488:511 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 0 3 theme High 70:73 arg1 Lipoprotein 83:93 High Density Lipoprotein 70:93 High Density Lipoprotein Particles 70:103 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 6 4 theme fetuin 921:926 arg1 antitrypsin 943:953 alpha 1 antitrypsin 935:953 alpha 1 antitrypsin 935:953 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 4 theme fetuin 921:926 arg1 glycoproteins 959:971 glycoproteins 959:971 glycoproteins associated with lipoproteins 959:1000 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 4 theme fetuin 921:926 arg1 CIII 915:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 4 theme fetuin 921:926 arg1 A 928:928 fetuin A 921:928 fetuin A 921:928 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 5 5 gly heterogeneity 859:871 arg1 CIII 837:840 Apo CIII 833:840 Apo CIII 833:840 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 1 6 theme High 166:169 arg1 HDL 192:194 HDL 192:194 HDL 192:194 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 1 6 theme High 166:169 arg1 lipoprotein 179:189 High density lipoprotein 166:189 High density lipoprotein (HDL) particles 166:205 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 7 7 from glycovariations 1174:1188 arg1 plasma 1215:1220 human plasma 1209:1220 human plasma 1209:1220 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 8 8 gly glycosylated 1345:1356 arg1 proteins 1297:1304 The proteins 1293:1304 The proteins found in the plasma 1293:1324 The proteins found in the plasma are differentially glycosylated compared to those isolated in HDL. 29212317 1 9 theme inverse 250:256 arg1 correlation 258:268 their inverse correlation 244:268 their inverse correlation with the prevalence of cardiovascular diseases 244:315 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 0 10 theme Lipoprotein 83:93 arg1 Particles 95:103 High Density Lipoprotein Particles 70:103 High Density Lipoprotein Particles 70:103 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 7 11 theme site-specific 1160:1172 arg1 glycovariations 1174:1188 site-specific glycovariations 1160:1188 site-specific glycovariations 1160:1188 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 3 12 theme HDL 548:550 arg1 constituents 552:563 HDL constituents 548:563 HDL constituents 548:563 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 9 13 theme functional 1514:1523 arg1 implications 1525:1536 possible functional implications 1505:1536 possible functional implications 1505:1536 The results of this study suggest that glycosylation may play a role in protein partitioning in the blood, with possible functional implications. 29212317 7 14 used used 1108:1111 arg2 UHPLC-QQQ 1093:1101 UHPLC-QQQ 1093:1101 UHPLC-QQQ 1093:1101 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 14 used used 1108:1111 arg2 method 1085:1090 A targeted method 1074:1090 A targeted method (UHPLC-QQQ) 1074:1102 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 6 15 theme Apolipoprotein 900:913 arg1 antitrypsin 943:953 alpha 1 antitrypsin 935:953 alpha 1 antitrypsin 935:953 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 15 theme Apolipoprotein 900:913 arg1 glycoproteins 959:971 glycoproteins 959:971 glycoproteins associated with lipoproteins 959:1000 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 15 theme Apolipoprotein 900:913 arg1 CIII 915:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 15 theme Apolipoprotein 900:913 arg1 A 928:928 fetuin A 921:928 fetuin A 921:928 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 7 16 dep concentrations 1141:1154 arg1 the 1124:1126 the 1124:1126 the 1124:1126 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 3 17 theme relative 525:532 arg1 abundances 534:543 the relative abundances 521:543 the relative abundances of HDL constituents 521:563 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 1 18 theme density 171:177 arg1 HDL 192:194 HDL 192:194 HDL 192:194 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 1 18 theme density 171:177 arg1 lipoprotein 179:189 High density lipoprotein 166:189 High density lipoprotein (HDL) particles 166:205 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 7 19 from proteins 1197:1204 arg1 plasma 1215:1220 human plasma 1209:1220 human plasma 1209:1220 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 5 20 theme HDL 777:779 arg1 associated-proteins 781:799 HDL associated-proteins 777:799 HDL associated-proteins 777:799 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 0 21 with Enriched 109:116 arg1 Glycosylation 132:144 Specific Glycosylation 123:144 Specific Glycosylation Compared to Plasma 123:163 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 5 22 gly glycosylation 760:772 arg1 associated-proteins 781:799 HDL associated-proteins 777:799 HDL associated-proteins 777:799 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 2 23 theme heart 379:383 arg1 disease 385:391 heart disease 379:391 heart disease 379:391 However, recent studies show that in some conditions such as heart disease and diabetes, HDL particles can become dysfunctional. 29212317 5 24 theme truncated 815:823 arg1 form 825:828 the truncated form 811:828 the truncated form of Apo CIII 811:840 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 7 25 theme HDL 1240:1242 arg1 particles 1244:1252 HDL particles 1240:1252 HDL particles isolated from the same plasma samples 1240:1290 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 26 theme proteins 1197:1204 arg1 concentrations 1141:1154 glycoprotein concentrations 1128:1154 glycoprotein concentrations 1128:1154 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 26 theme proteins 1197:1204 arg1 glycovariations 1174:1188 site-specific glycovariations 1160:1188 site-specific glycovariations 1160:1188 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 4 27 theme plasma 676:681 arg1 particles 683:691 plasma particles 676:691 plasma particles 676:691 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 0 28 theme Targeted 0:7 arg1 Measurements 9:20 Targeted Measurements 0:20 Targeted Measurements of O- and N-Glycopeptides 0:46 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 5 29 theme site-specific 878:890 arg1 manner 892:897 a site-specific manner 876:897 a site-specific manner 876:897 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 0 30 theme O- 25:26 arg1 Measurements 9:20 Targeted Measurements 0:20 Targeted Measurements of O- and N-Glycopeptides 0:46 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 0 31 theme Specific 123:130 arg1 Glycosylation 132:144 Specific Glycosylation 123:144 Specific Glycosylation Compared to Plasma 123:163 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 6 32 gly glycoproteins 959:971 arg1 antitrypsin 943:953 alpha 1 antitrypsin 935:953 alpha 1 antitrypsin 935:953 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 32 gly glycoproteins 959:971 arg1 glycoproteins 959:971 glycoproteins 959:971 glycoproteins associated with lipoproteins 959:1000 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 32 gly glycoproteins 959:971 arg1 CIII 915:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 32 gly glycoproteins 959:971 arg1 A 928:928 fetuin A 921:928 fetuin A 921:928 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 3 33 theme constituents 552:563 arg1 abundances 534:543 the relative abundances 521:543 the relative abundances of HDL constituents 521:563 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 2 34 theme HDL 407:409 arg1 particles 411:419 HDL particles 407:419 HDL particles 407:419 However, recent studies show that in some conditions such as heart disease and diabetes, HDL particles can become dysfunctional. 29212317 5 35 theme Apo 833:835 arg1 CIII 837:840 Apo CIII 833:840 Apo CIII 833:840 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 9 36 theme possible 1505:1512 arg1 implications 1525:1536 possible functional implications 1505:1536 possible functional implications 1505:1536 The results of this study suggest that glycosylation may play a role in protein partitioning in the blood, with possible functional implications. 29212317 5 37 from profile 732:738 arg1 manner 892:897 a site-specific manner 876:897 a site-specific manner 876:897 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 5 38 link N-linked 751:758 arg1 glycosylation 760:772 N-linked glycosylation 751:772 N-linked glycosylation 751:772 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 0 39 theme N-Glycopeptides 32:46 arg1 Measurements 9:20 Targeted Measurements 0:20 Targeted Measurements of O- and N-Glycopeptides 0:46 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 7 40 from plasma 1215:1220 arg1 concentrations 1141:1154 glycoprotein concentrations 1128:1154 glycoprotein concentrations 1128:1154 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 40 from plasma 1215:1220 arg1 glycovariations 1174:1188 site-specific glycovariations 1160:1188 site-specific glycovariations 1160:1188 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 4 41 dep structure 647:655 arg1 the 643:645 the 643:645 the 643:645 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 9 42 theme protein 1465:1471 arg1 partitioning 1473:1484 protein partitioning 1465:1484 protein partitioning in the blood 1465:1497 The results of this study suggest that glycosylation may play a role in protein partitioning in the blood, with possible functional implications. 29212317 6 43 theme disease 1054:1060 arg1 conditions 1062:1071 many cardiovascular and other disease conditions 1024:1071 many cardiovascular and other disease conditions 1024:1071 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 5 44 theme associated-proteins 781:799 arg1 O- 744:745 O- 744:745 O- 744:745 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 5 44 theme associated-proteins 781:799 arg1 glycosylation 760:772 N-linked glycosylation 751:772 N-linked glycosylation 751:772 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 1 45 theme cardiovascular 293:306 arg1 diseases 308:315 cardiovascular diseases 293:315 cardiovascular diseases 293:315 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 7 46 gly glycoprotein 1128:1139 arg1 glycoprotein 1128:1139 glycoprotein concentrations 1128:1154 glycoprotein concentrations 1128:1154 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 47 from concentrations 1141:1154 arg1 plasma 1215:1220 human plasma 1209:1220 human plasma 1209:1220 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 3 48 theme Great 447:451 arg1 attention 453:461 Great attention 447:461 Great attention 447:461 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 7 49 theme glycoprotein 1128:1139 arg1 concentrations 1141:1154 glycoprotein concentrations 1128:1154 glycoprotein concentrations 1128:1154 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 1 50 theme diseases 308:315 arg1 prevalence 279:288 the prevalence 275:288 the prevalence of cardiovascular diseases 275:315 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 6 51 theme other 1048:1052 arg1 conditions 1062:1071 many cardiovascular and other disease conditions 1024:1071 many cardiovascular and other disease conditions 1024:1071 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 0 52 from Proteins 58:65 arg1 Particles 95:103 High Density Lipoprotein Particles 70:103 High Density Lipoprotein Particles 70:103 Targeted Measurements of O- and N-Glycopeptides Show That Proteins in High Density Lipoprotein Particles Are Enriched with Specific Glycosylation Compared to Plasma. 29212317 3 53 theme functional 581:590 arg1 properties 592:601 HDL's functional properties 575:601 HDL's functional properties 575:601 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 1 54 with correlation 258:268 arg1 prevalence 279:288 the prevalence 275:288 the prevalence of cardiovascular diseases 275:315 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 7 55 theme targeted 1076:1083 arg1 UHPLC-QQQ 1093:1101 UHPLC-QQQ 1093:1101 UHPLC-QQQ 1093:1101 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 55 theme targeted 1076:1083 arg1 method 1085:1090 A targeted method 1074:1090 A targeted method (UHPLC-QQQ) 1074:1102 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 6 56 theme alpha 935:939 arg1 A 928:928 fetuin A 921:928 fetuin A 921:928 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 56 theme alpha 935:939 arg1 glycoproteins 959:971 glycoproteins 959:971 glycoproteins associated with lipoproteins 959:1000 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 56 theme alpha 935:939 arg1 antitrypsin 943:953 alpha 1 antitrypsin 935:953 alpha 1 antitrypsin 935:953 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 6 56 theme alpha 935:939 arg1 CIII 915:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII 900:918 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 5 57 dep O- 744:745 arg1 the 740:742 the 740:742 the 740:742 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 9 58 from partitioning 1473:1484 arg1 blood 1493:1497 the blood 1489:1497 the blood 1489:1497 The results of this study suggest that glycosylation may play a role in protein partitioning in the blood, with possible functional implications. 29212317 3 59 theme HDL 488:490 arg1 composition 501:511 HDL particle composition 488:511 HDL particle composition 488:511 Great attention has been directed toward HDL particle composition because the relative abundances of HDL constituents determine HDL's functional properties. 29212317 6 60 theme cardiovascular 1029:1042 arg1 conditions 1062:1071 many cardiovascular and other disease conditions 1024:1071 many cardiovascular and other disease conditions 1024:1071 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 2 61 theme recent 327:332 arg1 studies 334:340 recent studies 327:340 recent studies 327:340 However, recent studies show that in some conditions such as heart disease and diabetes, HDL particles can become dysfunctional. 29212317 4 62 dep factor 610:615 arg1 consider 620:627 consider 620:627 to consider when studying the structure and composition of plasma particles 617:691 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 7 63 attach isolated 1254:1261 arg2 particles 1244:1252 HDL particles 1240:1252 HDL particles isolated from the same plasma samples 1240:1290 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 63 attach isolated 1254:1261 arg1 samples 1284:1290 the same plasma samples 1268:1290 the same plasma samples 1268:1290 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 7 64 theme plasma 1277:1282 arg1 samples 1284:1290 the same plasma samples 1268:1290 the same plasma samples 1268:1290 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 4 65 theme key 606:608 arg1 factor 610:615 A key factor 604:615 A key factor to consider when studying the structure and composition of plasma particles 604:691 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 4 65 theme key 606:608 arg1 glycosylation 708:720 the protein glycosylation 696:720 the protein glycosylation 696:720 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 9 66 theme study 1413:1417 arg1 results 1397:1403 The results 1393:1403 The results of this study 1393:1417 The results of this study suggest that glycosylation may play a role in protein partitioning in the blood, with possible functional implications. 29212317 5 67 theme glycan 852:857 arg1 heterogeneity 859:871 their glycan heterogeneity 846:871 their glycan heterogeneity 846:871 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 5 68 theme CIII 837:840 arg1 form 825:828 the truncated form 811:828 the truncated form of Apo CIII 811:840 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 5 68 theme CIII 837:840 arg1 heterogeneity 859:871 their glycan heterogeneity 846:871 their glycan heterogeneity 846:871 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 7 69 theme same 1272:1275 arg1 samples 1284:1290 the same plasma samples 1268:1290 the same plasma samples 1268:1290 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 1 70 theme lipoprotein 179:189 arg1 particles 197:205 High density lipoprotein (HDL) particles 166:205 High density lipoprotein (HDL) particles 166:205 High density lipoprotein (HDL) particles are believed to be protective due to their inverse correlation with the prevalence of cardiovascular diseases. 29212317 6 71 theme many 1024:1027 arg1 conditions 1062:1071 many cardiovascular and other disease conditions 1024:1071 many cardiovascular and other disease conditions 1024:1071 Apolipoprotein CIII, fetuin A, and alpha 1 antitrypsin are glycoproteins associated with lipoproteins and are implicated in many cardiovascular and other disease conditions. 29212317 5 72 theme N-linked 751:758 arg1 glycosylation 760:772 N-linked glycosylation 751:772 N-linked glycosylation 751:772 Here, we profile the O- and N-linked glycosylation of HDL associated-proteins including the truncated form of Apo CIII and their glycan heterogeneity in a site-specific manner. 29212317 7 73 theme human 1209:1213 arg1 plasma 1215:1220 human plasma 1209:1220 human plasma 1209:1220 A targeted method (UHPLC-QQQ) was used to measure the glycoprotein concentrations and site-specific glycovariations of the proteins in human plasma and compared with HDL particles isolated from the same plasma samples. 29212317 4 74 theme particles 683:691 arg1 composition 661:671 composition 661:671 composition 661:671 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 4 74 theme particles 683:691 arg1 structure 647:655 structure 647:655 structure 647:655 A key factor to consider when studying the structure and composition of plasma particles is the protein glycosylation. 29212317 8 75 located found 1306:1310 arg2 proteins 1297:1304 The proteins 1293:1304 The proteins found in the plasma 1293:1324 The proteins found in the plasma are differentially glycosylated compared to those isolated in HDL. 29212317 8 75 located found 1306:1310 arg1 plasma 1319:1324 the plasma 1315:1324 the plasma 1315:1324 The proteins found in the plasma are differentially glycosylated compared to those isolated in HDL. 24509848 0 0 theme receptor 79:86 arg1 state 49:53 the activation state 34:53 the activation state of the discoidin domain receptor 1 (DDR1) 34:95 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 0 1 from Asn211 17:22 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation at Asn211 0:22 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 10 2 theme NDS 1527:1529 arg1 motif 1531:1535 the highly conserved (211)NDS motif 1501:1535 the highly conserved (211)NDS motif 1501:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 11 3 theme unwarranted 1750:1760 arg1 signaling 1762:1770 unwarranted signaling 1750:1770 unwarranted signaling by receptor dimers 1750:1789 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 8 4 theme similar 1246:1252 arg1 affinity 1237:1244 an affinity 1234:1244 an affinity similar to that of the wild-type DDR1 ectodomain 1234:1293 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 0 5 theme domain 72:77 arg1 receptor 79:86 discoidin domain receptor 1 62:88 the discoidin domain receptor 1 (DDR1) 58:95 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 0 5 theme domain 72:77 arg1 DDR1 91:94 DDR1 91:94 DDR1 91:94 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 6 6 theme NDS 858:860 arg1 motif 878:882 the conserved (211)NDS N-glycosylation motif 839:882 the conserved (211)NDS N-glycosylation motif 839:882 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 7 7 theme Mass 1020:1023 arg1 spectrometry 1025:1036 Mass spectrometry 1020:1036 Mass spectrometry 1020:1036 Mass spectrometry revealed that the N211Q mutant undergoes phosphorylation at Tyr(484), Tyr(520), Tyr(792), and Tyr(797). 24509848 6 8 theme site-directed 775:787 arg1 mutagenesis 789:799 site-directed mutagenesis 775:799 site-directed mutagenesis 775:799 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 2 9 theme sustained 310:318 arg1 kinetics 320:327 a slow and sustained kinetics 299:327 a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family 299:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 2 9 theme sustained 310:318 arg1 unique 337:342 unique 337:342 unique 337:342 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 10 10 theme conserved 1512:1520 arg1 motif 1531:1535 the highly conserved (211)NDS motif 1501:1535 the highly conserved (211)NDS motif 1501:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 9 11 theme wild-type 1316:1324 arg1 receptor 1326:1333 the wild-type receptor 1312:1333 the wild-type receptor 1312:1333 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 12 12 theme novel 1816:1820 arg1 insight 1822:1828 a novel insight 1814:1828 a novel insight into the structural mechanisms that regulate DDR activation 1814:1888 These studies provide a novel insight into the structural mechanisms that regulate DDR activation. 24509848 11 13 theme inactive 1723:1730 arg1 state 1732:1736 the inactive state 1719:1736 the inactive state 1719:1736 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 8 14 theme wild-type 1269:1277 arg1 ectodomain 1284:1293 the wild-type DDR1 ectodomain 1265:1293 the wild-type DDR1 ectodomain 1265:1293 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 4 15 theme autoinhibitory 575:588 arg1 state 590:594 the autoinhibitory state 571:594 the autoinhibitory state of the DDR1 dimers 571:613 However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. 24509848 10 16 theme DDR1 1579:1582 arg1 phosphorylation 1584:1598 DDR1 phosphorylation 1579:1598 DDR1 phosphorylation 1579:1598 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 11 17 from domain 1709:1714 arg1 state 1732:1736 the inactive state 1719:1736 the inactive state 1719:1736 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 11 18 from site 1666:1669 arg1 presence 1630:1637 The presence 1626:1637 The presence of glycan moieties at that site 1626:1669 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 9 19 theme N211Q 1340:1344 arg1 mutant 1346:1351 the N211Q mutant 1336:1351 the N211Q mutant 1336:1351 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 6 20 theme constitutive 990:1001 arg1 phosphorylation 1003:1017 collagen I-independent constitutive phosphorylation 967:1017 collagen I-independent constitutive phosphorylation 967:1017 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 12 21 theme DDR 1875:1877 arg1 activation 1879:1888 DDR activation 1875:1888 DDR activation 1875:1888 These studies provide a novel insight into the structural mechanisms that regulate DDR activation. 24509848 11 22 theme glycan 1642:1647 arg1 moieties 1649:1656 glycan moieties 1642:1656 glycan moieties 1642:1656 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 4 23 theme dimers 608:613 arg1 state 590:594 the autoinhibitory state 571:594 the autoinhibitory state of the DDR1 dimers 571:613 However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. 24509848 8 24 theme cell 1168:1171 arg1 surface 1173:1179 the cell surface 1164:1179 the cell surface 1164:1179 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 3 25 theme DDR1 398:401 arg1 dimerization 403:414 DDR1 dimerization 398:414 DDR1 dimerization 398:414 DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. 24509848 5 26 theme unique 696:701 arg1 role 703:706 a unique role 694:706 a unique role 694:706 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 6 27 theme conserved 843:851 arg1 motif 878:882 the conserved (211)NDS N-glycosylation motif 839:882 the conserved (211)NDS N-glycosylation motif 839:882 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 5 28 theme autophosphorylation 748:766 arg1 control 715:721 the control 711:721 the control of DDR1 dimerization and autophosphorylation 711:766 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 6 29 theme N-glycosylation 899:913 arg1 Asn 946:948 Asn(394) 946:953 Asn(394) 946:953 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 29 theme N-glycosylation 899:913 arg1 sites 915:919 other N-glycosylation sites 893:919 other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)) 893:954 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 29 theme N-glycosylation 899:913 arg1 Asn 932:934 Asn(371) 932:939 Asn(371) 932:939 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 29 theme N-glycosylation 899:913 arg1 Asn 922:924 Asn(260) 922:929 Asn(260) 922:929 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 9 30 theme enhanced 1362:1369 arg1 dimerization 1380:1391 enhanced receptor dimerization 1362:1391 enhanced receptor dimerization 1362:1391 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 8 31 theme DDR1 1279:1282 arg1 ectodomain 1284:1293 the wild-type DDR1 ectodomain 1265:1293 the wild-type DDR1 ectodomain 1265:1293 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 6 32 gly N-glycosylation 899:913 arg2 Asn 922:924 Asn(260) 922:929 Asn(260) 922:929 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 32 gly N-glycosylation 899:913 arg2 sites 915:919 other N-glycosylation sites 893:919 other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)) 893:954 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 32 gly N-glycosylation 899:913 arg2 Asn 932:934 Asn(371) 932:939 Asn(371) 932:939 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 32 gly N-glycosylation 899:913 arg2 Asn 946:948 Asn(394) 946:953 Asn(394) 946:953 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 2 33 theme collagen 277:284 arg1 binding 286:292 collagen binding 277:292 collagen binding 277:292 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 1 34 theme unique 146:151 arg1 family 153:158 a unique family 144:158 a unique family of receptor tyrosine kinases that signal in response to collagens 144:224 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 7 35 theme N211Q 1056:1060 arg1 mutant 1062:1067 the N211Q mutant 1052:1067 the N211Q mutant 1052:1067 Mass spectrometry revealed that the N211Q mutant undergoes phosphorylation at Tyr(484), Tyr(520), Tyr(792), and Tyr(797). 24509848 4 36 theme mechanism 543:551 arg1 unknown 618:624 unknown 618:624 unknown 618:624 However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. 24509848 4 36 theme mechanism 543:551 arg1 s 553:553 the mechanism(s) 539:554 the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers 539:613 However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. 24509848 5 37 theme Asn 671:673 arg1 residue 680:686 the Asn(211) residue 667:686 the Asn(211) residue 667:686 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 9 38 theme ligand 1423:1428 arg1 withdrawal 1430:1439 ligand withdrawal 1423:1439 ligand withdrawal 1423:1439 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 9 39 theme sustained 1397:1405 arg1 activation 1407:1416 sustained activation 1397:1416 sustained activation 1397:1416 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 2 40 theme kinase 383:388 arg1 family 390:395 the receptor tyrosine kinase family 361:395 the receptor tyrosine kinase family 361:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 2 41 theme family 390:395 arg1 members 350:356 members 350:356 members of the receptor tyrosine kinase family 350:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 11 42 attach presence 1630:1637 arg1 site 1666:1669 that site 1661:1669 that site 1661:1669 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 11 42 attach presence 1630:1637 arg2 moieties 1649:1656 glycan moieties 1642:1656 glycan moieties 1642:1656 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 9 43 theme receptor 1371:1378 arg1 dimerization 1380:1391 enhanced receptor dimerization 1362:1391 enhanced receptor dimerization 1362:1391 However, unlike the wild-type receptor, the N211Q mutant exhibits enhanced receptor dimerization and sustained activation upon ligand withdrawal. 24509848 6 44 theme N-glycosylation 862:876 arg1 motif 878:882 the conserved (211)NDS N-glycosylation motif 839:882 the conserved (211)NDS N-glycosylation motif 839:882 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 2 45 theme tyrosine 374:381 arg1 family 390:395 the receptor tyrosine kinase family 361:395 the receptor tyrosine kinase family 361:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 0 46 theme activation 38:47 arg1 state 49:53 the activation state 34:53 the activation state of the discoidin domain receptor 1 (DDR1) 34:95 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 2 47 theme slow 301:304 arg1 kinetics 320:327 a slow and sustained kinetics 299:327 a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family 299:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 2 47 theme slow 301:304 arg1 unique 337:342 unique 337:342 unique 337:342 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 10 48 theme phosphorylation 1584:1598 arg1 repressor 1566:1574 a negative repressor 1555:1574 a negative repressor of DDR1 phosphorylation 1555:1598 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 10 48 theme phosphorylation 1584:1598 arg1 N-glycosylation 1482:1496 N-glycosylation 1482:1496 N-glycosylation at the highly conserved (211)NDS motif 1482:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 1 49 theme receptor 163:170 arg1 kinases 181:187 receptor tyrosine kinases 163:187 receptor tyrosine kinases that signal in response to collagens 163:224 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 0 50 gly Glycosylation 0:12 arg2 Asn211 17:22 Asn211 17:22 Asn211 17:22 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 2 51 theme receptor 365:372 arg1 family 390:395 the receptor tyrosine kinase family 361:395 the receptor tyrosine kinase family 361:395 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 5 52 theme DDR1 726:729 arg1 dimerization 731:742 DDR1 dimerization 726:742 DDR1 dimerization 726:742 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 6 53 theme other 893:897 arg1 Asn 946:948 Asn(394) 946:953 Asn(394) 946:953 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 53 theme other 893:897 arg1 sites 915:919 other N-glycosylation sites 893:919 other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)) 893:954 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 53 theme other 893:897 arg1 Asn 932:934 Asn(371) 932:939 Asn(371) 932:939 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 53 theme other 893:897 arg1 Asn 922:924 Asn(260) 922:929 Asn(260) 922:929 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 5 54 from residue 680:686 arg1 N-glycosylation 648:662 N-glycosylation 648:662 N-glycosylation at the Asn(211) residue 648:686 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 3 55 theme receptor 425:432 arg1 activation 434:443 receptor activation 425:443 receptor activation 425:443 DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. 24509848 5 56 theme dimerization 731:742 arg1 control 715:721 the control 711:721 the control of DDR1 dimerization and autophosphorylation 711:766 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 10 57 gly N-glycosylation 1482:1496 arg2 motif 1531:1535 the highly conserved (211)NDS motif 1501:1535 the highly conserved (211)NDS motif 1501:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 10 57 gly N-glycosylation 1482:1496 arg1 motif 1531:1535 the highly conserved (211)NDS motif 1501:1535 the highly conserved (211)NDS motif 1501:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 8 58 theme I 1219:1219 arg1 binding 1221:1227 collagen I binding 1210:1227 collagen I binding 1210:1227 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 6 59 theme I-independent 976:988 arg1 phosphorylation 1003:1017 collagen I-independent constitutive phosphorylation 967:1017 collagen I-independent constitutive phosphorylation 967:1017 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 3 60 theme inhibitory 469:478 arg1 mechanism 480:488 a structural inhibitory mechanism 456:488 a structural inhibitory mechanism to prevent unwarranted phosphorylation 456:527 DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. 24509848 10 61 dep conserved 1512:1520 arg1 211 1523:1525 211 1523:1525 211 1523:1525 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 11 62 theme moieties 1649:1656 arg1 presence 1630:1637 The presence 1626:1637 The presence of glycan moieties at that site 1626:1669 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 5 63 gly N-glycosylation 648:662 arg2 residue 680:686 the Asn(211) residue 667:686 the Asn(211) residue 667:686 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 5 63 gly N-glycosylation 648:662 arg1 residue 680:686 the Asn(211) residue 667:686 the Asn(211) residue 667:686 Here, we report that N-glycosylation at the Asn(211) residue plays a unique role in the control of DDR1 dimerization and autophosphorylation. 24509848 6 64 dep sites 915:919 arg1 Asn 946:948 Asn(394) 946:953 Asn(394) 946:953 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 64 dep sites 915:919 arg1 sites 915:919 other N-glycosylation sites 893:919 other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)) 893:954 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 64 dep sites 915:919 arg1 Asn 932:934 Asn(371) 932:939 Asn(371) 932:939 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 64 dep sites 915:919 arg1 Asn 922:924 Asn(260) 922:929 Asn(260) 922:929 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 8 65 theme collagen 1210:1217 arg1 binding 1221:1227 collagen I binding 1210:1227 collagen I binding 1210:1227 The N211Q traffics to the cell surface, and its ectodomain displays collagen I binding with an affinity similar to that of the wild-type DDR1 ectodomain. 24509848 1 66 theme Discoidin 98:106 arg1 receptor 115:122 Discoidin domain receptor 1 98:124 Discoidin domain receptor 1 (DDR1) 98:131 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 1 66 theme Discoidin 98:106 arg1 DDR1 127:130 DDR1 127:130 DDR1 127:130 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 10 67 theme negative 1557:1564 arg1 repressor 1566:1574 a negative repressor 1555:1574 a negative repressor of DDR1 phosphorylation 1555:1598 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 10 67 theme negative 1557:1564 arg1 N-glycosylation 1482:1496 N-glycosylation 1482:1496 N-glycosylation at the highly conserved (211)NDS motif 1482:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 10 68 from motif 1531:1535 arg1 N-glycosylation 1482:1496 N-glycosylation 1482:1496 N-glycosylation at the highly conserved (211)NDS motif 1482:1535 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 10 68 from motif 1531:1535 arg1 repressor 1566:1574 a negative repressor 1555:1574 a negative repressor of DDR1 phosphorylation 1555:1598 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 11 69 theme collagen-binding 1692:1707 arg1 domain 1709:1714 the collagen-binding domain 1688:1714 the collagen-binding domain in the inactive state 1688:1736 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 1 70 theme domain 108:113 arg1 receptor 115:122 Discoidin domain receptor 1 98:124 Discoidin domain receptor 1 (DDR1) 98:131 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 1 70 theme domain 108:113 arg1 DDR1 127:130 DDR1 127:130 DDR1 127:130 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 1 71 theme kinases 181:187 arg1 family 153:158 a unique family 144:158 a unique family of receptor tyrosine kinases that signal in response to collagens 144:224 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 11 72 theme receptor 1775:1782 arg1 dimers 1784:1789 receptor dimers 1775:1789 receptor dimers 1775:1789 The presence of glycan moieties at that site may help to lock the collagen-binding domain in the inactive state and prevent unwarranted signaling by receptor dimers. 24509848 3 73 theme structural 458:467 arg1 mechanism 480:488 a structural inhibitory mechanism 456:488 a structural inhibitory mechanism to prevent unwarranted phosphorylation 456:527 DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. 24509848 2 74 dep binding 286:292 arg1 response 265:272 response 265:272 response 265:272 DDR1 undergoes autophosphorylation in response to collagen binding with a slow and sustained kinetics that is unique among members of the receptor tyrosine kinase family. 24509848 0 75 theme discoidin 62:70 arg1 receptor 79:86 discoidin domain receptor 1 62:88 the discoidin domain receptor 1 (DDR1) 58:95 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 0 75 theme discoidin 62:70 arg1 DDR1 91:94 DDR1 91:94 DDR1 91:94 Glycosylation at Asn211 regulates the activation state of the discoidin domain receptor 1 (DDR1). 24509848 1 76 theme tyrosine 172:179 arg1 kinases 181:187 receptor tyrosine kinases 163:187 receptor tyrosine kinases that signal in response to collagens 163:224 Discoidin domain receptor 1 (DDR1) belongs to a unique family of receptor tyrosine kinases that signal in response to collagens. 24509848 10 77 theme ligand 1618:1623 arg1 absence 1607:1613 the absence 1603:1613 the absence of ligand 1603:1623 Taken together, these data suggest that N-glycosylation at the highly conserved (211)NDS motif evolved to act as a negative repressor of DDR1 phosphorylation in the absence of ligand. 24509848 3 78 theme unwarranted 501:511 arg1 phosphorylation 513:527 unwarranted phosphorylation 501:527 unwarranted phosphorylation 501:527 DDR1 dimerization precedes receptor activation suggesting a structural inhibitory mechanism to prevent unwarranted phosphorylation. 24509848 4 79 theme DDR1 603:606 arg1 dimers 608:613 the DDR1 dimers 599:613 the DDR1 dimers 599:613 However, the mechanism(s) that maintains the autoinhibitory state of the DDR1 dimers is unknown. 24509848 12 80 theme structural 1839:1848 arg1 mechanisms 1850:1859 the structural mechanisms 1835:1859 the structural mechanisms that regulate DDR activation 1835:1888 These studies provide a novel insight into the structural mechanisms that regulate DDR activation. 24509848 6 81 theme collagen 967:974 arg1 phosphorylation 1003:1017 collagen I-independent constitutive phosphorylation 967:1017 collagen I-independent constitutive phosphorylation 967:1017 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 24509848 6 82 gly N-glycosylation 862:876 arg2 motif 878:882 the conserved (211)NDS N-glycosylation motif 839:882 the conserved (211)NDS N-glycosylation motif 839:882 Using site-directed mutagenesis, we found that mutations that disrupt the conserved (211)NDS N-glycosylation motif, but not other N-glycosylation sites (Asn(260), Asn(371), and Asn(394)), result in collagen I-independent constitutive phosphorylation. 26348906 10 0 theme transport 1703:1711 arg1 activity 1713:1720 transport activity 1703:1720 transport activity 1703:1720 Substitution by alanine or valine at either site caused a dramatic loss in transport activity. 26348906 6 1 theme homology 725:732 arg1 modelling 734:742 homology modelling 725:742 homology modelling 725:742 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 2 theme Xenopus 1025:1031 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 3 3 theme Re-establishing 457:471 arg1 isotonicity 473:483 Re-establishing isotonicity 457:483 Re-establishing isotonicity 457:483 Re-establishing isotonicity involves PM depletion of BGT-1. 26348906 10 4 from site 1672:1675 arg1 Substitution 1628:1639 Substitution 1628:1639 Substitution by alanine or valine at either site 1628:1675 Substitution by alanine or valine at either site caused a dramatic loss in transport activity. 26348906 11 5 theme SLC6 1880:1883 arg1 transporter 1885:1895 this SLC6 transporter 1875:1895 this SLC6 transporter 1875:1895 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 6 6 theme Madin-Darby 1097:1107 arg1 kidney 1116:1121 Madin-Darby canine kidney 1097:1121 Madin-Darby canine kidney 1097:1121 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 6 theme Madin-Darby 1097:1107 arg1 MDCK 1091:1094 MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1091:1164 MDCK 1091:1094 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 3 7 theme BGT-1 510:514 arg1 depletion 497:505 PM depletion 494:505 PM depletion of BGT-1 494:514 Re-establishing isotonicity involves PM depletion of BGT-1. 26348906 6 8 theme voltage 995:1001 arg1 measurements 1009:1020 two-electrode voltage clamp measurements 981:1020 two-electrode voltage clamp measurements 981:1020 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 5 9 theme present 632:638 arg1 study 640:644 the present study 628:644 the present study 628:644 In the present study, we reveal a link between regulated PM insertion and N-glycosylation. 26348906 1 10 theme symporter 164:172 arg1 member 212:217 a member 210:217 a member of the SLC6 (solute carrier 6) family 210:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 10 theme symporter 164:172 arg1 BGT-1 174:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 113:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1) 113:207 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 10 theme symporter 164:172 arg1 transporter 194:204 betaine/GABA transporter 1 181:206 betaine/GABA transporter 1 181:206 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 7 11 gly N-glycosylation 1229:1243 arg1 oocytes 1253:1259 oocytes 1253:1259 oocytes 1253:1259 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 7 11 gly N-glycosylation 1229:1243 arg1 cells 1270:1274 MDCK cells 1265:1274 MDCK cells 1265:1274 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 9 12 theme activity 1491:1498 arg1 loss 1477:1480 no loss 1474:1480 no loss of BGT-1 activity 1474:1498 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 9 13 theme double 1513:1518 arg1 inactive 1531:1538 inactive 1531:1538 inactive 1531:1538 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 9 13 theme double 1513:1518 arg1 mutant 1520:1525 the double mutant 1509:1525 the double mutant 1509:1525 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 6 14 theme radiolabelled 1062:1074 arg1 substrate 1076:1084 radiolabelled substrate 1062:1084 radiolabelled substrate 1062:1084 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 7 15 theme BGT-1 1199:1203 arg1 insertion 1186:1194 Trafficking and PM insertion 1167:1194 Trafficking and PM insertion of BGT-1 1167:1203 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 6 16 theme embryonic 1142:1150 arg1 cells 1160:1164 MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1091:1164 cells 1160:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 16 theme embryonic 1142:1150 arg1 kidney 1152:1157 human embryonic kidney 1136:1157 human embryonic kidney 1136:1157 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 11 17 theme PM 1750:1751 arg1 insertion 1753:1761 PM insertion 1750:1761 PM insertion of N183D 1750:1770 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 9 18 gly N-glycosylation 1557:1571 arg1 one 1585:1587 one 1585:1587 one 1585:1587 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 9 18 gly N-glycosylation 1557:1571 arg1 sites 1596:1600 the sites 1592:1600 the sites 1592:1600 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 1 19 theme Na 123:124 arg1 member 212:217 a member 210:217 a member of the SLC6 (solute carrier 6) family 210:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 19 theme Na 123:124 arg1 BGT-1 174:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 113:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1) 113:207 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 19 theme Na 123:124 arg1 transporter 194:204 betaine/GABA transporter 1 181:206 betaine/GABA transporter 1 181:206 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 2 20 theme elevated 420:427 arg1 betaine 429:435 elevated betaine 420:435 elevated betaine 420:435 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 5 21 theme regulated 672:680 arg1 insertion 685:693 regulated PM insertion 672:693 regulated PM insertion 672:693 In the present study, we reveal a link between regulated PM insertion and N-glycosylation. 26348906 11 22 theme MDCK 1739:1742 arg1 cells 1744:1748 MDCK cells 1739:1748 MDCK cells 1739:1748 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 1 23 theme -coupled 128:135 arg1 GABA 137:140 -coupled GABA 128:140 -coupled GABA 128:140 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 0 24 from Role 0:3 arg1 transport 41:49 renal betaine transport 27:49 renal betaine transport 27:49 Role of N-glycosylation in renal betaine transport. 26348906 6 25 theme BGT-1 830:834 arg1 loop 814:817 the extracellular loop 2 796:819 the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 796:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 25 theme BGT-1 830:834 arg1 EL2 822:824 EL2 822:824 EL2 822:824 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 2 26 theme membrane 329:336 arg1 insertion 343:351 the transcription, translation and plasma membrane (PM) insertion 287:351 the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells 287:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 1 27 theme folding 69:75 arg1 betaine 87:93 The osmolyte and folding chaperone betaine 52:93 betaine 87:93 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 11 28 theme osmotic 1799:1805 arg1 stress 1807:1812 osmotic stress 1799:1812 osmotic stress 1799:1812 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 7 29 theme PM 1183:1184 arg1 insertion 1186:1194 Trafficking and PM insertion 1167:1194 Trafficking and PM insertion of BGT-1 1167:1203 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 2 30 theme kidney 365:370 arg1 cells 372:376 kidney cells 365:376 kidney cells 365:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 8 31 from Asn 1330:1332 arg1 association 1287:1297 association 1287:1297 association with N-glycans at Asn(171) and Asn(183) 1287:1337 Moreover, association with N-glycans at Asn(171) and Asn(183) contributed equally to protein activity and substrate affinity. 26348906 7 32 theme Trafficking 1167:1177 arg1 insertion 1186:1194 Trafficking and PM insertion 1167:1194 Trafficking and PM insertion of BGT-1 1167:1203 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 2 33 theme BGT-1 356:360 arg1 insertion 343:351 the transcription, translation and plasma membrane (PM) insertion 287:351 the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells 287:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 0 34 theme N-glycosylation 8:22 arg1 Role 0:3 Role 0:3 Role of N-glycosylation in renal betaine transport. 0:50 Role of N-glycosylation in renal betaine transport. 26348906 7 35 theme MDCK 1265:1268 arg1 cells 1270:1274 MDCK cells 1265:1274 MDCK cells 1265:1274 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 9 36 theme sites 1596:1600 arg1 one 1585:1587 one 1585:1587 one 1585:1587 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 9 36 theme sites 1596:1600 arg1 sites 1596:1600 the sites 1592:1600 the sites 1592:1600 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 0 37 theme renal 27:31 arg1 transport 41:49 renal betaine transport 27:49 renal betaine transport 27:49 Role of N-glycosylation in renal betaine transport. 26348906 9 38 theme Asn 1419:1421 arg1 Substitution 1403:1414 Substitution 1403:1414 Substitution of Asn(171) and Asn(183) by aspartate 1403:1452 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 8 39 theme substrate 1383:1391 arg1 affinity 1393:1400 substrate affinity 1383:1400 substrate affinity 1383:1400 Moreover, association with N-glycans at Asn(171) and Asn(183) contributed equally to protein activity and substrate affinity. 26348906 8 40 with association 1287:1297 arg1 N-glycans 1304:1312 N-glycans 1304:1312 N-glycans 1304:1312 Moreover, association with N-glycans at Asn(171) and Asn(183) contributed equally to protein activity and substrate affinity. 26348906 1 41 theme betaine/GABA 181:192 arg1 BGT-1 174:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 113:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1) 113:207 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 41 theme betaine/GABA 181:192 arg1 transporter 194:204 betaine/GABA transporter 1 181:206 betaine/GABA transporter 1 181:206 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 2 42 from insertion 343:351 arg1 cells 372:376 kidney cells 365:376 kidney cells 365:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 4 43 theme regulated 548:556 arg1 insertion 561:569 the regulated PM insertion 544:569 the regulated PM insertion of BGT-1 544:578 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 6 44 theme human 1136:1140 arg1 cells 1160:1164 MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1091:1164 cells 1160:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 44 theme human 1136:1140 arg1 kidney 1152:1157 human embryonic kidney 1136:1157 human embryonic kidney 1136:1157 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 45 theme electron 939:946 arg1 EM 960:961 EM 960:961 EM 960:961 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 45 theme electron 939:946 arg1 microscopy 948:957 electron microscopy 939:957 electron microscopy (EM) 939:962 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 46 theme HEK293 1128:1133 arg1 cells 1160:1164 MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1091:1164 cells 1160:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 46 theme HEK293 1128:1133 arg1 kidney 1152:1157 human embryonic kidney 1136:1157 human embryonic kidney 1136:1157 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 47 from transport 904:912 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 11 48 theme N183D 1766:1770 arg1 insertion 1753:1761 PM insertion 1750:1761 PM insertion of N183D 1750:1770 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 6 49 theme laevis 1033:1038 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 4 50 theme molecular 521:529 arg1 mechanism 531:539 The molecular mechanism 517:539 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress 517:611 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 4 50 theme molecular 521:529 arg1 unknown 616:622 unknown 616:622 unknown 616:622 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 1 51 theme GABA 137:140 arg1 member 212:217 a member 210:217 a member of the SLC6 (solute carrier 6) family 210:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 51 theme GABA 137:140 arg1 BGT-1 174:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 113:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1) 113:207 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 51 theme GABA 137:140 arg1 transporter 194:204 betaine/GABA transporter 1 181:206 betaine/GABA transporter 1 181:206 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 10 52 theme dramatic 1686:1693 arg1 loss 1695:1698 a dramatic loss 1684:1698 a dramatic loss in transport activity 1684:1720 Substitution by alanine or valine at either site caused a dramatic loss in transport activity. 26348906 6 53 theme canine 1109:1114 arg1 kidney 1116:1121 Madin-Darby canine kidney 1097:1121 Madin-Darby canine kidney 1097:1121 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 53 theme canine 1109:1114 arg1 MDCK 1091:1094 MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1091:1164 MDCK 1091:1094 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 54 from trafficking 877:887 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 4 55 from changes 587:593 arg1 stress 606:611 osmotic stress 598:611 osmotic stress 598:611 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 1 56 theme γ-aminobutyric 143:156 arg1 acid 158:161 γ-aminobutyric acid 143:161 γ-aminobutyric acid 143:161 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 56 theme γ-aminobutyric 143:156 arg1 Na 123:124 renal Na 117:124 renal Na 117:124 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 11 57 theme transporter 1885:1895 arg1 regulation 1861:1870 regulation 1861:1870 regulation of this SLC6 transporter 1861:1895 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 6 58 theme clamp 1003:1007 arg1 measurements 1009:1020 two-electrode voltage clamp measurements 981:1020 two-electrode voltage clamp measurements 981:1020 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 4 59 theme osmotic 598:604 arg1 stress 606:611 osmotic stress 598:611 osmotic stress 598:611 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 6 60 from insertion 890:898 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 3 61 theme PM 494:495 arg1 depletion 497:505 PM depletion 494:505 PM depletion of BGT-1 494:514 Re-establishing isotonicity involves PM depletion of BGT-1. 26348906 9 62 theme BGT-1 1485:1489 arg1 activity 1491:1498 BGT-1 activity 1485:1498 BGT-1 activity 1485:1498 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 6 63 theme two-electrode 981:993 arg1 measurements 1009:1020 two-electrode voltage clamp measurements 981:1020 two-electrode voltage clamp measurements 981:1020 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 4 64 theme insertion 561:569 arg1 mechanism 531:539 The molecular mechanism 517:539 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress 517:611 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 4 64 theme insertion 561:569 arg1 unknown 616:622 unknown 616:622 unknown 616:622 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 4 65 theme BGT-1 574:578 arg1 insertion 561:569 the regulated PM insertion 544:569 the regulated PM insertion of BGT-1 544:578 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 6 66 theme extracellular 800:812 arg1 loop 814:817 the extracellular loop 2 796:819 the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 796:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 66 theme extracellular 800:812 arg1 EL2 822:824 EL2 822:824 EL2 822:824 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 11 67 theme N-glycosylation 1842:1856 arg1 impact 1832:1837 the impact 1828:1837 the impact of N-glycosylation in regulation of this SLC6 transporter 1828:1895 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 6 68 theme substrate 1076:1084 arg1 insertion 890:898 insertion 890:898 insertion 890:898 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 68 theme substrate 1076:1084 arg1 trafficking 877:887 trafficking 877:887 trafficking 877:887 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 68 theme substrate 1076:1084 arg1 uptake 1052:1057 uptake 1052:1057 uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells 1052:1164 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 68 theme substrate 1076:1084 arg1 transport 904:912 transport 904:912 transport 904:912 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 1 69 theme renal 117:121 arg1 Na 123:124 renal Na 117:124 renal Na 117:124 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 69 theme renal 117:121 arg1 acid 158:161 γ-aminobutyric acid 143:161 γ-aminobutyric acid 143:161 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 11 70 from impact 1832:1837 arg1 regulation 1861:1870 regulation 1861:1870 regulation of this SLC6 transporter 1861:1895 Furthermore, in MDCK cells PM insertion of N183D was no longer regulated by osmotic stress, highlighting the impact of N-glycosylation in regulation of this SLC6 transporter. 26348906 6 71 from uptake 1052:1057 arg1 oocytes 1040:1046 Xenopus laevis oocytes 1025:1046 Xenopus laevis oocytes 1025:1046 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 5 72 theme PM 682:683 arg1 insertion 685:693 regulated PM insertion 672:693 regulated PM insertion 672:693 In the present study, we reveal a link between regulated PM insertion and N-glycosylation. 26348906 2 73 theme GABA 441:444 arg1 transport 446:454 GABA transport 441:454 GABA transport 441:454 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 8 74 from Asn 1317:1319 arg1 association 1287:1297 association 1287:1297 association with N-glycans at Asn(171) and Asn(183) 1287:1337 Moreover, association with N-glycans at Asn(171) and Asn(183) contributed equally to protein activity and substrate affinity. 26348906 10 75 from loss 1695:1698 arg1 activity 1713:1720 transport activity 1703:1720 transport activity 1703:1720 Substitution by alanine or valine at either site caused a dramatic loss in transport activity. 26348906 2 76 theme plasma 322:327 arg1 PM 339:340 PM 339:340 PM 339:340 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 2 76 theme plasma 322:327 arg1 membrane 329:336 plasma membrane 322:336 plasma membrane (PM) 322:341 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 1 77 theme chaperone 77:85 arg1 betaine 87:93 The osmolyte and folding chaperone betaine 52:93 betaine 87:93 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 9 78 theme one 1585:1587 arg1 N-glycosylation 1557:1571 N-glycosylation 1557:1571 N-glycosylation of at least one of the sites 1557:1600 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 8 79 theme protein 1362:1368 arg1 activity 1370:1377 protein activity 1362:1377 protein activity 1362:1377 Moreover, association with N-glycans at Asn(171) and Asn(183) contributed equally to protein activity and substrate affinity. 26348906 2 80 theme translation 306:316 arg1 insertion 343:351 the transcription, translation and plasma membrane (PM) insertion 287:351 the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells 287:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 1 81 theme SLC6 226:229 arg1 carrier 239:245 solute carrier 6 232:247 solute carrier 6 232:247 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 81 theme SLC6 226:229 arg1 family 250:255 the SLC6 (solute carrier 6) family 222:255 the SLC6 (solute carrier 6) family 222:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 6 82 dep sites 763:767 arg1 Asn 783:785 Asn(183) 783:790 Asn(183) 783:790 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 82 dep sites 763:767 arg1 Asn 770:772 Asn(171) 770:777 Asn(171) 770:777 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 6 82 dep sites 763:767 arg1 sites 763:767 two sites 759:767 two sites (Asn(171) and Asn(183)) 759:791 Based on homology modelling, we identified two sites (Asn(171) and Asn(183)) in the extracellular loop 2 (EL2) of BGT-1, which were investigated with respect to trafficking, insertion and transport by immunogold-labelling, electron microscopy (EM), mutagenesis and two-electrode voltage clamp measurements in Xenopus laevis oocytes and uptake of radiolabelled substrate into MDCK (Madin-Darby canine kidney) and HEK293 (human embryonic kidney) cells. 26348906 0 83 theme betaine 33:39 arg1 transport 41:49 renal betaine transport 27:49 renal betaine transport 27:49 Role of N-glycosylation in renal betaine transport. 26348906 1 84 theme solute 232:237 arg1 carrier 239:245 solute carrier 6 232:247 solute carrier 6 232:247 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 84 theme solute 232:237 arg1 family 250:255 the SLC6 (solute carrier 6) family 222:255 the SLC6 (solute carrier 6) family 222:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 7 85 from N-glycosylation 1229:1243 arg1 oocytes 1253:1259 oocytes 1253:1259 oocytes 1253:1259 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 7 85 from N-glycosylation 1229:1243 arg1 cells 1270:1274 MDCK cells 1265:1274 MDCK cells 1265:1274 Trafficking and PM insertion of BGT-1 was clearly promoted by N-glycosylation in both oocytes and MDCK cells. 26348906 0 86 gly N-glycosylation 8:22 arg1 betaine 33:39 renal betaine transport 27:49 renal betaine transport 27:49 Role of N-glycosylation in renal betaine transport. 26348906 2 87 theme hypertonic 264:273 arg1 conditions 275:284 hypertonic conditions 264:284 hypertonic conditions 264:284 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 26348906 1 88 theme family 250:255 arg1 BGT-1 174:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 113:178 the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1) 113:207 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 1 88 theme family 250:255 arg1 member 212:217 a member 210:217 a member of the SLC6 (solute carrier 6) family 210:255 The osmolyte and folding chaperone betaine is transported by the renal Na(+)-coupled GABA (γ-aminobutyric acid) symporter BGT-1 (betaine/GABA transporter 1), a member of the SLC6 (solute carrier 6) family. 26348906 9 89 theme Asn 1432:1434 arg1 Substitution 1403:1414 Substitution 1403:1414 Substitution of Asn(171) and Asn(183) by aspartate 1403:1452 Substitution of Asn(171) and Asn(183) by aspartate individually caused no loss of BGT-1 activity, whereas the double mutant was inactive, suggesting that N-glycosylation of at least one of the sites is required for function. 26348906 4 90 theme PM 558:559 arg1 insertion 561:569 the regulated PM insertion 544:569 the regulated PM insertion of BGT-1 544:578 The molecular mechanism of the regulated PM insertion of BGT-1 during changes in osmotic stress is unknown. 26348906 2 91 theme transcription 291:303 arg1 insertion 343:351 the transcription, translation and plasma membrane (PM) insertion 287:351 the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells 287:376 Under hypertonic conditions, the transcription, translation and plasma membrane (PM) insertion of BGT-1 in kidney cells are significantly increased, resulting in elevated betaine and GABA transport. 25202310 8 0 theme central 2067:2073 arg1 lectins 2059:2065 lectins 2059:2065 lectins central to infection and immunity 2059:2099 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 4 1 theme glycome 762:768 arg1 profiling 770:778 Detailed subcellular glycome profiling 741:778 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) 741:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 6 2 theme subcellular-specific 1650:1669 arg1 N-glycosylation 1671:1685 subcellular-specific N-glycosylation 1650:1685 subcellular-specific N-glycosylation 1650:1685 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 3 3 theme complex 664:670 arg1 N-glycans 678:686 N-glycans 678:686 N-glycans 678:686 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 3 3 theme complex 664:670 arg1 type 672:675 complex type 664:675 complex type 664:675 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 3 4 theme high-mannose-rich 697:713 arg1 glycoproteins 726:738 the high-mannose-rich microsomal glycoproteins 693:738 the high-mannose-rich microsomal glycoproteins 693:738 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 1 5 theme innate 168:173 arg1 immunity 188:195 innate and adaptive immunity 168:195 innate and adaptive immunity 168:195 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 2 6 gly glycoproteins 375:387 arg1 glycoproteins 375:387 glycoproteins 375:387 glycoproteins trafficking through the shared biosynthetic machinery of human cells 375:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 5 7 theme gene 1135:1138 arg1 ontology 1140:1147 gene ontology 1135:1147 gene ontology 1135:1147 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 7 8 theme shared 1830:1835 arg1 machinery 1850:1858 a shared biosynthetic machinery 1828:1858 a shared biosynthetic machinery 1828:1858 In conclusion, human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery. 25202310 6 9 from accessibility 1378:1390 arg1 proteins 1438:1445 maturely folded proteins 1422:1445 maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1422:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 2 10 theme biosynthetic 420:431 arg1 machinery 433:441 the shared biosynthetic machinery 409:441 the shared biosynthetic machinery of human cells 409:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 6 11 theme putative 1472:1479 arg1 N-glycoproteins 1481:1495 the 100 most abundant putative N-glycoproteins 1450:1495 the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1450:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 4 12 theme cell 818:821 arg1 lines 823:827 three breast cell lines 805:827 three breast cell lines (MCF7/MDA468/MCF10A) 805:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 4 12 theme cell 818:821 arg1 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 7 13 theme subcellular-specific 1785:1804 arg1 N-glycosylation 1806:1820 subcellular-specific N-glycosylation 1785:1820 subcellular-specific N-glycosylation using a shared biosynthetic machinery 1785:1858 In conclusion, human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery. 25202310 4 14 attach derived 792:798 arg1 lines 823:827 three breast cell lines 805:827 three breast cell lines (MCF7/MDA468/MCF10A) 805:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 4 14 attach derived 792:798 arg2 proteins 783:790 proteins 783:790 proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) 783:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 4 14 attach derived 792:798 arg1 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 8 15 theme glycosylation 1893:1905 arg1 aspect 1866:1871 This aspect 1861:1871 This aspect of protein-specific glycosylation 1861:1905 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 6 16 theme glycan 1575:1580 arg1 processing 1587:1596 the glycan type processing 1571:1596 the glycan type processing 1571:1596 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 4 17 gly glycoproteins 877:889 arg1 glycoproteins 877:889 secreted glycoproteins 868:889 secreted glycoproteins 868:889 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 5 18 theme resident 1204:1211 arg1 glycoproteins 1213:1225 endoplasmic reticulum resident glycoproteins 1182:1225 endoplasmic reticulum resident glycoproteins 1182:1225 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 3 19 gly glycoproteins 726:738 arg1 glycoproteins 726:738 the high-mannose-rich microsomal glycoproteins 693:738 the high-mannose-rich microsomal glycoproteins 693:738 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 2 20 theme N-glycosylation 343:357 arg1 determinants 359:370 subcellular-specific N-glycosylation determinants 322:370 subcellular-specific N-glycosylation determinants 322:370 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 1 21 theme adaptive 179:186 arg1 immunity 188:195 innate and adaptive immunity 168:195 innate and adaptive immunity 168:195 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 8 22 theme structural 1924:1933 arg1 glycobiology 1950:1961 structural and functional glycobiology 1924:1961 structural and functional glycobiology 1924:1961 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 2 23 dep glycoproteins 375:387 arg1 trafficking 389:399 trafficking 389:399 trafficking 389:399 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 1 24 theme lectin-based 200:211 arg1 interactions 213:224 lectin-based interactions 200:224 lectin-based interactions to exposed glyco-determinants 200:254 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 8 25 theme interaction 2020:2030 arg1 context 1989:1995 the context 1985:1995 the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity 1985:2099 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 5 26 theme endoplasmic 1182:1192 arg1 reticulum 1194:1202 endoplasmic reticulum 1182:1202 endoplasmic reticulum resident glycoproteins 1182:1225 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 3 27 theme LC-MS/MS-based 459:472 arg1 glycomics 487:495 LC-MS/MS-based quantitative glycomics 459:495 LC-MS/MS-based quantitative glycomics 459:495 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 1 28 theme extra- 130:135 arg1 functions 155:163 extra- and intracellular functions 130:163 extra- and intracellular functions 130:163 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 2 29 from formation 309:317 arg1 glycoproteins 375:387 glycoproteins 375:387 glycoproteins trafficking through the shared biosynthetic machinery of human cells 375:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 6 30 theme glycosylation 1399:1411 arg1 sites 1413:1417 the glycosylation sites 1395:1417 the glycosylation sites 1395:1417 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 3 31 theme secreted 513:520 arg1 glycoproteins 522:534 the secreted glycoproteins 509:534 the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes 509:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 3 32 theme epithelial 558:567 arg1 cells 569:573 eight human breast epithelial cells 539:573 eight human breast epithelial cells displaying diverse geno- and phenotypes 539:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 0 33 theme Differential 0:11 arg1 accessibility 18:30 Differential site accessibility 0:30 Differential site accessibility 0:30 Differential site accessibility mechanistically explains subcellular-specific N-glycosylation determinants. 25202310 1 34 theme intracellular 141:153 arg1 functions 155:163 extra- and intracellular functions 130:163 extra- and intracellular functions 130:163 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 6 35 theme solvent 1370:1376 arg1 accessibility 1378:1390 The solvent accessibility 1366:1390 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1366:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 3 36 theme human 545:549 arg1 cells 569:573 eight human breast epithelial cells 539:573 eight human breast epithelial cells displaying diverse geno- and phenotypes 539:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 8 37 theme spatio-temporal 2004:2018 arg1 interaction 2020:2030 the spatio-temporal interaction 2000:2030 the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity 2000:2099 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 5 38 theme respective 1332:1341 arg1 fractions 1355:1363 the respective subcellular fractions 1328:1363 the respective subcellular fractions 1328:1363 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 6 39 gly glycosylation 1399:1411 arg2 sites 1413:1417 the glycosylation sites 1395:1417 the glycosylation sites 1395:1417 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 4 40 theme microsomal 1081:1090 arg1 glycoproteomes 1092:1105 the under-processed microsomal glycoproteomes 1061:1105 the under-processed microsomal glycoproteomes 1061:1105 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 0 41 theme N-glycosylation 78:92 arg1 determinants 94:105 subcellular-specific N-glycosylation determinants 57:105 subcellular-specific N-glycosylation determinants 57:105 Differential site accessibility mechanistically explains subcellular-specific N-glycosylation determinants. 25202310 4 42 theme glycan-processed 1012:1027 arg1 glycoproteomes 1042:1055 the intermediately glycan-processed cell-surface glycoproteomes 993:1055 the intermediately glycan-processed cell-surface glycoproteomes 993:1055 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 2 43 theme human 446:450 arg1 cells 452:456 human cells 446:456 human cells 446:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 5 44 theme cell-surface 1298:1309 arg1 glycoproteins 1311:1323 secreted and cell-surface glycoproteins 1285:1323 glycoproteins 1311:1323 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 4 45 theme proteins 783:790 arg1 profiling 770:778 Detailed subcellular glycome profiling 741:778 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) 741:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 6 46 theme folded 1431:1436 arg1 proteins 1438:1445 maturely folded proteins 1422:1445 maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1422:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 6 47 theme N-glycosylation 1671:1685 arg1 formation 1637:1645 the formation 1633:1645 the formation of subcellular-specific N-glycosylation 1633:1685 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 8 48 theme functional 1939:1948 arg1 glycobiology 1950:1961 structural and functional glycobiology 1924:1961 structural and functional glycobiology 1924:1961 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 4 49 theme subcellular 750:760 arg1 profiling 770:778 Detailed subcellular glycome profiling 741:778 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) 741:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 5 50 from enrichment 1271:1280 arg1 fractions 1355:1363 the respective subcellular fractions 1328:1363 the respective subcellular fractions 1328:1363 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 5 51 attach presence 1170:1177 arg2 glycoproteins 1213:1225 endoplasmic reticulum resident glycoproteins 1182:1225 endoplasmic reticulum resident glycoproteins 1182:1225 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 5 51 attach presence 1170:1177 arg1 microsomes 1234:1243 the microsomes 1230:1243 the microsomes 1230:1243 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 1 52 theme exposed 229:235 arg1 glyco-determinants 237:254 exposed glyco-determinants 229:254 exposed glyco-determinants 229:254 Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. 25202310 6 53 gly N-glycoproteins 1481:1495 arg1 N-glycoproteins 1481:1495 the 100 most abundant putative N-glycoproteins 1450:1495 the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1450:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 7 54 theme biosynthetic 1837:1848 arg1 machinery 1850:1858 a shared biosynthetic machinery 1828:1858 a shared biosynthetic machinery 1828:1858 In conclusion, human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery. 25202310 2 55 theme shared 413:418 arg1 machinery 433:441 the shared biosynthetic machinery 409:441 the shared biosynthetic machinery of human cells 409:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 6 56 theme N-glycoproteins 1481:1495 arg1 proteins 1438:1445 maturely folded proteins 1422:1445 maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1422:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 8 57 theme protein-specific 1876:1891 arg1 glycosylation 1893:1905 protein-specific glycosylation 1876:1905 protein-specific glycosylation 1876:1905 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 6 58 theme abundant 1463:1470 arg1 N-glycoproteins 1481:1495 the 100 most abundant putative N-glycoproteins 1450:1495 the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1450:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 4 59 theme breast 811:816 arg1 lines 823:827 three breast cell lines 805:827 three breast cell lines (MCF7/MDA468/MCF10A) 805:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 4 59 theme breast 811:816 arg1 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 MCF7/MDA468/MCF10A 830:847 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 8 60 with interaction 2020:2030 arg1 lectins 2059:2065 lectins 2059:2065 lectins central to infection and immunity 2059:2099 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 5 61 theme Subcellular 1108:1118 arg1 proteomics 1120:1129 Subcellular proteomics 1108:1129 Subcellular proteomics 1108:1129 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 5 62 gly glycoproteins 1213:1225 arg1 glycoproteins 1213:1225 endoplasmic reticulum resident glycoproteins 1182:1225 endoplasmic reticulum resident glycoproteins 1182:1225 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 6 63 theme type 1582:1585 arg1 processing 1587:1596 the glycan type processing 1571:1596 the glycan type processing 1571:1596 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 5 64 theme reticulum 1194:1202 arg1 glycoproteins 1213:1225 endoplasmic reticulum resident glycoproteins 1182:1225 endoplasmic reticulum resident glycoproteins 1182:1225 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 2 65 theme determinants 359:370 arg1 formation 309:317 the formation 305:317 the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells 305:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 5 66 theme glycoproteins 1213:1225 arg1 presence 1170:1177 substantial presence 1158:1177 substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes 1158:1243 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 3 67 theme microsomal 715:724 arg1 glycoproteins 726:738 the high-mannose-rich microsomal glycoproteins 693:738 the high-mannose-rich microsomal glycoproteins 693:738 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 2 68 theme subcellular-specific 322:341 arg1 determinants 359:370 subcellular-specific N-glycosylation determinants 322:370 subcellular-specific N-glycosylation determinants 322:370 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 5 69 theme substantial 1158:1168 arg1 presence 1170:1177 substantial presence 1158:1177 substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes 1158:1243 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 6 70 theme subcellular 1528:1538 arg1 glycoproteomes 1540:1553 the three subcellular glycoproteomes 1518:1553 the three subcellular glycoproteomes 1518:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 4 71 theme secreted 868:875 arg1 glycoproteins 877:889 secreted glycoproteins 868:889 secreted glycoproteins 868:889 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 5 72 theme significant 1259:1269 arg1 enrichment 1271:1280 significant enrichment 1259:1280 significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions 1259:1363 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 3 73 gly glycoproteins 522:534 arg1 glycoproteins 522:534 the secreted glycoproteins 509:534 the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes 509:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 5 74 from presence 1170:1177 arg1 microsomes 1234:1243 the microsomes 1230:1243 the microsomes 1230:1243 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 6 75 theme sites 1413:1417 arg1 accessibility 1378:1390 The solvent accessibility 1366:1390 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1366:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 3 76 theme quantitative 474:485 arg1 glycomics 487:495 LC-MS/MS-based quantitative glycomics 459:495 LC-MS/MS-based quantitative glycomics 459:495 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 6 77 located observed 1497:1504 arg1 glycoproteomes 1540:1553 the three subcellular glycoproteomes 1518:1553 the three subcellular glycoproteomes 1518:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 6 77 located observed 1497:1504 arg2 N-glycoproteins 1481:1495 the 100 most abundant putative N-glycoproteins 1450:1495 the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes 1450:1553 The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. 25202310 0 78 theme site 13:16 arg1 accessibility 18:30 Differential site accessibility 0:30 Differential site accessibility 0:30 Differential site accessibility mechanistically explains subcellular-specific N-glycosylation determinants. 25202310 7 79 theme human 1703:1707 arg1 cells 1709:1713 human cells 1703:1713 human cells 1703:1713 In conclusion, human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery. 25202310 3 80 theme cells 569:573 arg1 glycoproteins 522:534 the secreted glycoproteins 509:534 the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes 509:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 3 81 dep type 672:675 arg1 glycoproteins 726:738 the high-mannose-rich microsomal glycoproteins 693:738 the high-mannose-rich microsomal glycoproteins 693:738 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 4 82 theme Detailed 741:748 arg1 profiling 770:778 Detailed subcellular glycome profiling 741:778 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) 741:848 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 0 83 theme subcellular-specific 57:76 arg1 determinants 94:105 subcellular-specific N-glycosylation determinants 57:105 subcellular-specific N-glycosylation determinants 57:105 Differential site accessibility mechanistically explains subcellular-specific N-glycosylation determinants. 25202310 3 84 theme breast 551:556 arg1 cells 569:573 eight human breast epithelial cells 539:573 eight human breast epithelial cells displaying diverse geno- and phenotypes 539:613 LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high-mannose-rich microsomal glycoproteins. 25202310 8 85 theme glyco-determinants 2035:2052 arg1 interaction 2020:2030 the spatio-temporal interaction 2000:2030 the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity 2000:2099 This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity. 25202310 5 86 theme subcellular 1343:1353 arg1 fractions 1355:1363 the respective subcellular fractions 1328:1363 the respective subcellular fractions 1328:1363 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 5 87 gly glycoproteins 1311:1323 arg1 glycoproteins 1311:1323 secreted and cell-surface glycoproteins 1285:1323 glycoproteins 1311:1323 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 4 88 theme under-processed 1065:1079 arg1 glycoproteomes 1092:1105 the under-processed microsomal glycoproteomes 1061:1105 the under-processed microsomal glycoproteomes 1061:1105 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25202310 2 89 theme cells 452:456 arg1 machinery 433:441 the shared biosynthetic machinery 409:441 the shared biosynthetic machinery of human cells 409:456 Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. 25202310 5 90 theme secreted 1285:1292 arg1 glycoproteins 1311:1323 secreted and cell-surface glycoproteins 1285:1323 glycoproteins 1311:1323 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 5 91 theme glycoproteins 1311:1323 arg1 enrichment 1271:1280 significant enrichment 1259:1280 significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions 1259:1363 Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell-surface glycoproteins in the respective subcellular fractions. 25202310 4 92 theme cell-surface 1029:1040 arg1 glycoproteomes 1042:1055 the intermediately glycan-processed cell-surface glycoproteomes 993:1055 the intermediately glycan-processed cell-surface glycoproteomes 993:1055 Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell-surface glycoproteomes and the under-processed microsomal glycoproteomes. 25528754 7 0 theme O-linked 607:614 arg1 glycans 616:622 O-linked glycans 607:622 O-linked glycans that are only present in the secreted DII 607:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 4 1 theme ligand-binding 400:413 arg1 portion 415:421 the ligand-binding portion 396:421 the ligand-binding portion of LDL receptor (LDLR) 396:444 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 11 2 theme novel 1126:1130 arg1 feature 1132:1138 a novel feature 1124:1138 a novel feature 1124:1138 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 12 3 theme LDL 1308:1310 arg1 retention 1312:1320 the early LDL retention 1298:1320 the early LDL retention 1298:1320 Thus, perlecan glycosylation may play a role in the early LDL retention during the development of atherosclerosis. 25528754 1 4 from proteoglycan 150:161 arg1 wall 179:182 the arterial wall 166:182 the arterial wall 166:182 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 12 5 theme perlecan 1256:1263 arg1 glycosylation 1265:1277 perlecan glycosylation 1256:1277 perlecan glycosylation 1256:1277 Thus, perlecan glycosylation may play a role in the early LDL retention during the development of atherosclerosis. 25528754 2 6 attach linked 207:212 arg2 studies 194:200 Previous studies 185:200 Previous studies 185:200 Previous studies have linked it to atherosclerosis. 25528754 2 6 attach linked 207:212 arg1 atherosclerosis 220:234 atherosclerosis 220:234 atherosclerosis 220:234 Previous studies have linked it to atherosclerosis. 25528754 7 7 from present 638:644 arg1 DII 662:664 the secreted DII 649:664 the secreted DII 649:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 10 8 from effect 1008:1013 arg1 binding 1036:1042 LDL binding 1032:1042 LDL binding 1032:1042 We further demonstrate an additive effect of HS and DII on LDL binding. 25528754 8 9 theme glycosylation 715:727 arg1 sites 729:733 the glycosylation sites 711:733 the glycosylation sites 711:733 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 11 10 theme LDLR-like 1156:1164 arg1 DII 1166:1168 the perlecan LDLR-like DII 1143:1168 the perlecan LDLR-like DII in receptor-mediated lipoprotein retention 1143:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 5 11 theme functional 451:460 arg1 significance 462:473 The functional significance 447:473 The functional significance of this domain 447:488 The functional significance of this domain has been unknown. 25528754 5 11 theme functional 451:460 arg1 unknown 499:505 unknown 499:505 unknown 499:505 The functional significance of this domain has been unknown. 25528754 9 12 theme charged 915:921 arg1 critical 947:954 critical 947:954 critical 947:954 Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. 25528754 9 12 theme charged 915:921 arg1 acids 930:934 the negatively charged sialic acids 900:934 the negatively charged sialic acids 900:934 Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. 25528754 6 13 theme perlecan 527:534 arg1 DII 536:538 perlecan DII 527:538 perlecan DII 527:538 Here, we show that perlecan DII interacts with LDL. 25528754 9 14 theme LDL 960:962 arg1 binding 964:970 LDL binding 960:970 LDL binding 960:970 Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. 25528754 0 15 theme glycosylation-dependent 4:26 arg1 interaction 28:38 The glycosylation-dependent interaction 0:38 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 0:107 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 11 16 from retention 1203:1211 arg1 feature 1132:1138 a novel feature 1124:1138 a novel feature 1124:1138 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 1 17 theme arterial 170:177 arg1 wall 179:182 the arterial wall 166:182 the arterial wall 166:182 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 11 18 theme LDL 1072:1074 arg1 uptake 1076:1081 LDL uptake 1072:1081 LDL uptake 1072:1081 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 11 19 theme perlecan 1147:1154 arg1 DII 1166:1168 the perlecan LDLR-like DII 1143:1168 the perlecan LDLR-like DII in receptor-mediated lipoprotein retention 1143:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 12 20 theme atherosclerosis 1348:1362 arg1 development 1333:1343 the development 1329:1343 the development of atherosclerosis 1329:1362 Thus, perlecan glycosylation may play a role in the early LDL retention during the development of atherosclerosis. 25528754 8 21 from serine 799:804 arg1 rich 791:794 rich 791:794 rich 791:794 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 4 22 theme disparate 343:351 arg1 structures 353:362 disparate structures 343:362 disparate structures 343:362 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 8 23 theme cysteine 832:839 arg1 residues 841:848 no cysteine residues 829:848 no cysteine residues 829:848 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 10 24 theme additive 999:1006 arg1 effect 1008:1013 an additive effect 996:1013 an additive effect of HS and DII on LDL binding 996:1042 We further demonstrate an additive effect of HS and DII on LDL binding. 25528754 7 25 from DII 662:664 arg1 present 638:644 present 638:644 present 638:644 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 0 26 theme core 52:55 arg1 protein 57:63 perlecan core protein 43:63 perlecan core protein 43:63 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 11 27 from DII 1166:1168 arg1 retention 1203:1211 receptor-mediated lipoprotein retention 1173:1211 receptor-mediated lipoprotein retention 1173:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 3 28 theme core 257:260 arg1 protein 262:268 a core protein 255:268 a core protein 255:268 Perlecan contains a core protein and three HS side chains. 25528754 7 29 located present 638:644 arg1 DII 662:664 the secreted DII 649:664 the secreted DII 649:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 7 29 located present 638:644 arg2 glycans 616:622 O-linked glycans 607:622 O-linked glycans that are only present in the secreted DII 607:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 8 30 from rich 791:794 arg1 serine 799:804 serine 799:804 serine 799:804 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 8 30 from rich 791:794 arg1 threonine 810:818 threonine 810:818 threonine 810:818 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 9 31 theme sialic 923:928 arg1 critical 947:954 critical 947:954 critical 947:954 Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. 25528754 9 31 theme sialic 923:928 arg1 acids 930:934 the negatively charged sialic acids 900:934 the negatively charged sialic acids 900:934 Interestingly, most of the glycans are capped by the negatively charged sialic acids, which are critical for LDL binding. 25528754 7 32 attach present 638:644 arg1 DII 662:664 the secreted DII 649:664 the secreted DII 649:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 7 32 attach present 638:644 arg2 glycans 616:622 O-linked glycans 607:622 O-linked glycans that are only present in the secreted DII 607:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 0 33 theme perlecan 43:50 arg1 protein 57:63 perlecan core protein 43:63 perlecan core protein 43:63 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 8 34 contain has 825:827 arg2 residues 841:848 no cysteine residues 829:848 no cysteine residues 829:848 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 8 34 contain has 825:827 arg1 divergent 777:785 divergent 777:785 divergent 777:785 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 8 34 contain has 825:827 arg1 unit 755:758 the second unit 744:758 the second unit 744:758 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 4 35 theme receptor 430:437 arg1 portion 415:421 the ligand-binding portion 396:421 the ligand-binding portion of LDL receptor (LDLR) 396:444 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 10 36 theme HS 1018:1019 arg1 effect 1008:1013 an additive effect 996:1013 an additive effect of HS and DII on LDL binding 996:1042 We further demonstrate an additive effect of HS and DII on LDL binding. 25528754 10 37 theme DII 1025:1027 arg1 effect 1008:1013 an additive effect 996:1013 an additive effect of HS and DII on LDL binding 996:1042 We further demonstrate an additive effect of HS and DII on LDL binding. 25528754 4 38 theme LDL 426:428 arg1 LDLR 440:443 LDLR 440:443 LDLR 440:443 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 4 38 theme LDL 426:428 arg1 receptor 430:437 LDL receptor 426:437 LDL receptor (LDLR) 426:444 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 0 39 with interaction 28:38 arg1 LDL 70:72 LDL 70:72 LDL 70:72 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 0 40 theme protein 57:63 arg1 interaction 28:38 The glycosylation-dependent interaction 0:38 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 0:107 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 11 41 from feature 1132:1138 arg1 retention 1203:1211 receptor-mediated lipoprotein retention 1173:1211 receptor-mediated lipoprotein retention 1173:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 11 42 theme DII 1166:1168 arg1 feature 1132:1138 a novel feature 1124:1138 a novel feature 1124:1138 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 1 43 theme major 123:127 arg1 HS 146:147 HS 146:147 HS 146:147 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 1 43 theme major 123:127 arg1 sulfate 137:143 major heparan sulfate 123:143 a major heparan sulfate (HS) proteoglycan in the arterial wall 121:182 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 7 44 link O-linked 607:614 arg1 glycans 616:622 O-linked glycans 607:622 O-linked glycans that are only present in the secreted DII 607:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 8 45 theme repeat 682:687 arg1 units 689:693 the five repeat units 673:693 the five repeat units of DII 673:700 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 3 46 theme HS 280:281 arg1 chains 288:293 three HS side chains 274:293 three HS side chains 274:293 Perlecan contains a core protein and three HS side chains. 25528754 8 47 from threonine 810:818 arg1 rich 791:794 rich 791:794 rich 791:794 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 1 48 theme heparan 129:135 arg1 HS 146:147 HS 146:147 HS 146:147 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 1 48 theme heparan 129:135 arg1 sulfate 137:143 major heparan sulfate 123:143 a major heparan sulfate (HS) proteoglycan in the arterial wall 121:182 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 8 49 theme DII 698:700 arg1 units 689:693 the five repeat units 673:693 the five repeat units of DII 673:700 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 3 50 theme side 283:286 arg1 chains 288:293 three HS side chains 274:293 three HS side chains 274:293 Perlecan contains a core protein and three HS side chains. 25528754 2 51 theme Previous 185:192 arg1 studies 194:200 Previous studies 185:200 Previous studies 185:200 Previous studies have linked it to atherosclerosis. 25528754 0 52 dep interaction 28:38 arg1 implications 75:86 implications 75:86 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 0:107 The glycosylation-dependent interaction of perlecan core protein with LDL: implications for atherosclerosis. 25528754 4 53 theme core 300:303 arg1 protein 305:311 Its core protein 296:311 Its core protein 296:311 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 1 54 theme sulfate 137:143 arg1 Perlecan 109:116 Perlecan 109:116 Perlecan 109:116 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 1 54 theme sulfate 137:143 arg1 proteoglycan 150:161 a major heparan sulfate (HS) proteoglycan 121:161 a major heparan sulfate (HS) proteoglycan in the arterial wall 121:182 Perlecan is a major heparan sulfate (HS) proteoglycan in the arterial wall. 25528754 8 55 theme second 748:753 arg1 divergent 777:785 divergent 777:785 divergent 777:785 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 8 55 theme second 748:753 arg1 unit 755:758 the second unit 744:758 the second unit 744:758 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 8 56 gly glycosylation 715:727 arg2 sites 729:733 the glycosylation sites 711:733 the glycosylation sites 711:733 Among the five repeat units of DII, most of the glycosylation sites are from the second unit, which is highly divergent and rich in serine and threonine, but has no cysteine residues. 25528754 5 57 theme domain 483:488 arg1 significance 462:473 The functional significance 447:473 The functional significance of this domain 447:488 The functional significance of this domain has been unknown. 25528754 5 57 theme domain 483:488 arg1 unknown 499:505 unknown 499:505 unknown 499:505 The functional significance of this domain has been unknown. 25528754 7 58 theme secreted 653:660 arg1 DII 662:664 the secreted DII 649:664 the secreted DII 649:664 Importantly, the interaction largely relies on O-linked glycans that are only present in the secreted DII. 25528754 3 59 contain contains 246:253 arg1 Perlecan 237:244 Perlecan 237:244 Perlecan 237:244 Perlecan contains a core protein and three HS side chains. 25528754 3 59 contain contains 246:253 arg2 chains 288:293 three HS side chains 274:293 three HS side chains 274:293 Perlecan contains a core protein and three HS side chains. 25528754 3 59 contain contains 246:253 arg2 protein 262:268 a core protein 255:268 a core protein 255:268 Perlecan contains a core protein and three HS side chains. 25528754 4 60 contain has 313:315 arg1 protein 305:311 Its core protein 296:311 Its core protein 296:311 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 4 60 contain has 313:315 arg2 DI-DV 331:335 DI-DV 331:335 DI-DV 331:335 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 4 60 contain has 313:315 arg2 domains 322:328 five domains 317:328 five domains (DI-DV) 317:336 Its core protein has five domains (DI-DV) with disparate structures and DII is highly homologous to the ligand-binding portion of LDL receptor (LDLR). 25528754 12 61 theme early 1302:1306 arg1 retention 1312:1320 the early LDL retention 1298:1320 the early LDL retention 1298:1320 Thus, perlecan glycosylation may play a role in the early LDL retention during the development of atherosclerosis. 25528754 11 62 theme receptor-mediated 1173:1189 arg1 retention 1203:1211 receptor-mediated lipoprotein retention 1173:1211 receptor-mediated lipoprotein retention 1173:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 11 63 theme lipoprotein 1191:1201 arg1 retention 1203:1211 receptor-mediated lipoprotein retention 1173:1211 receptor-mediated lipoprotein retention 1173:1211 Unlike LDLR, which directs LDL uptake through endocytosis, this study uncovers a novel feature of the perlecan LDLR-like DII in receptor-mediated lipoprotein retention, which depends on its glycosylation. 25528754 10 64 theme LDL 1032:1034 arg1 binding 1036:1042 LDL binding 1032:1042 LDL binding 1032:1042 We further demonstrate an additive effect of HS and DII on LDL binding. 25916169 2 0 gly N-glycosylation 318:332 arg2 Asn 341:343 Asn 341:343 Asn(92) 341:347 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 0 gly N-glycosylation 318:332 arg2 sites 334:338 two putative N-glycosylation sites 305:338 two putative N-glycosylation sites (Asn(92) and Asn(211)) 305:361 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 0 gly N-glycosylation 318:332 arg2 Asn 353:355 Asn 353:355 Asn(211) 353:360 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 0 gly N-glycosylation 318:332 arg2 two 305:307 two 305:307 two 305:307 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 5 1 theme proximal 709:716 arg1 compartments 718:729 proximal compartments 709:729 proximal compartments 709:729 We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. 25916169 1 2 theme transmembrane 165:177 arg1 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 2 theme transmembrane 165:177 arg1 protein 179:185 a type II transmembrane protein 155:185 a type II transmembrane protein 155:185 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 9 3 theme N-terminal 1343:1352 arg1 retain 1522:1527 retain 1522:1527 to retain it in the Golgi complex 1519:1551 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 3 theme N-terminal 1343:1352 arg1 able 1449:1452 able 1449:1452 able 1449:1452 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 3 theme N-terminal 1343:1352 arg1 domain 1364:1369 the N-terminal ST3Gal-II domain 1339:1369 the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) 1339:1444 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 3 theme N-terminal 1343:1352 arg1 drive 1457:1461 drive 1457:1461 to drive a protein reporter out of the endoplasmic reticulum 1454:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 0 4 theme intracellular 73:85 arg1 trafficking 87:97 intracellular trafficking 73:97 intracellular trafficking 73:97 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 5 from role 9:12 arg1 activity 103:110 activity 103:110 activity 103:110 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 5 from role 9:12 arg1 trafficking 87:97 intracellular trafficking 73:97 intracellular trafficking 73:97 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 5 from role 9:12 arg1 Asn211 59:64 Asn211 59:64 Asn211 59:64 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 8 6 from Asn 1115:1117 arg1 N-glycan 1094:1101 the N-glycan 1090:1101 the N-glycan at position Asn(211) 1090:1122 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 9 7 theme ST3Gal-II 1354:1362 arg1 retain 1522:1527 retain 1522:1527 to retain it in the Golgi complex 1519:1551 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 7 theme ST3Gal-II 1354:1362 arg1 able 1449:1452 able 1449:1452 able 1449:1452 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 7 theme ST3Gal-II 1354:1362 arg1 domain 1364:1369 the N-terminal ST3Gal-II domain 1339:1369 the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) 1339:1444 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 7 theme ST3Gal-II 1354:1362 arg1 drive 1457:1461 drive 1457:1461 to drive a protein reporter out of the endoplasmic reticulum 1454:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 6 8 from Asn 850:852 arg1 N-glycosylated 832:845 N-glycosylated 832:845 N-glycosylated 832:845 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 6 9 theme site-directed 768:780 arg1 mutagenesis 782:792 pharmacological, biochemical and site-directed mutagenesis 735:792 pharmacological, biochemical and site-directed mutagenesis 735:792 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 2 10 theme putative 309:316 arg1 sites 334:338 two putative N-glycosylation sites 305:338 two putative N-glycosylation sites (Asn(92) and Asn(211)) 305:361 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 10 theme putative 309:316 arg1 Asn 353:355 Asn 353:355 Asn(211) 353:360 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 10 theme putative 309:316 arg1 Asn 341:343 Asn 341:343 Asn(92) 341:347 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 9 11 dep able 1449:1452 arg1 retain 1522:1527 retain 1522:1527 to retain it in the Golgi complex 1519:1551 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 11 dep able 1449:1452 arg1 able 1449:1452 able 1449:1452 able 1449:1452 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 11 dep able 1449:1452 arg1 domain 1364:1369 the N-terminal ST3Gal-II domain 1339:1369 the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) 1339:1444 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 11 dep able 1449:1452 arg1 drive 1457:1461 drive 1457:1461 to drive a protein reporter out of the endoplasmic reticulum 1454:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 6 12 theme endoplasmic 937:947 arg1 reticulum 949:957 the endoplasmic reticulum 933:957 the endoplasmic reticulum 933:957 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 5 13 theme Golgi 684:688 arg1 complex 690:696 the Golgi complex 680:696 the Golgi complex 680:696 We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. 25916169 0 14 from Asn211 59:64 arg1 glycosylation 42:54 evolutionarily conserved glycosylation 17:54 evolutionarily conserved glycosylation at Asn211 17:64 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 14 from Asn211 59:64 arg1 role 9:12 Critical role 0:12 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 0:142 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 6 15 from localization 976:987 arg1 exit 923:926 its exit 919:926 its exit from the endoplasmic reticulum and proper Golgi localization 919:987 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 9 16 theme protein 1465:1471 arg1 reporter 1473:1480 a protein reporter 1463:1480 a protein reporter out of the endoplasmic reticulum 1463:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 10 17 theme ST3Gal-II 1598:1606 arg1 ST3Gal-II 1598:1606 ST3Gal-II 1598:1606 ST3Gal-II 1598:1606 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 10 17 theme ST3Gal-II 1598:1606 arg1 domain 1588:1593 the C-terminal domain 1573:1593 the C-terminal domain of ST3Gal-II 1573:1606 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 0 18 theme sialyltransferase 115:131 arg1 activity 103:110 activity 103:110 activity 103:110 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 18 theme sialyltransferase 115:131 arg1 trafficking 87:97 intracellular trafficking 73:97 intracellular trafficking 73:97 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 8 19 theme N-glycan 1255:1262 arg1 mutant 1264:1269 the single N-glycan mutant 1244:1269 the single N-glycan mutant at Asn(92) 1244:1280 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 6 20 theme biochemical 752:762 arg1 mutagenesis 782:792 pharmacological, biochemical and site-directed mutagenesis 735:792 pharmacological, biochemical and site-directed mutagenesis 735:792 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 8 21 theme moderate 1293:1300 arg1 effect 1302:1307 only a moderate effect 1286:1307 only a moderate effect 1286:1307 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 4 22 theme N-glycosylations 601:616 arg1 trafficking 550:560 intracellular trafficking 536:560 intracellular trafficking 536:560 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 4 22 theme N-glycosylations 601:616 arg1 activity 573:580 enzyme activity 566:580 enzyme activity 566:580 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 10 23 theme proper 1673:1678 arg1 exit 1680:1683 proper exit 1673:1683 proper exit from the endoplasmic reticulum 1673:1714 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 8 24 theme enzyme 1174:1179 arg1 activity 1158:1165 the activity 1154:1165 the activity of the enzyme 1154:1179 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 9 25 contain containing 1371:1380 arg1 retain 1522:1527 retain 1522:1527 to retain it in the Golgi complex 1519:1551 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 25 contain containing 1371:1380 arg1 able 1449:1452 able 1449:1452 able 1449:1452 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 25 contain containing 1371:1380 arg1 drive 1457:1461 drive 1457:1461 to drive a protein reporter out of the endoplasmic reticulum 1454:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 25 contain containing 1371:1380 arg2 region 1420:1425 the cytosolic, transmembrane and stem region 1382:1425 region 1420:1425 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 25 contain containing 1371:1380 arg1 domain 1364:1369 the N-terminal ST3Gal-II domain 1339:1369 the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) 1339:1444 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 25 contain containing 1371:1380 arg2 acids 1434:1438 amino acids 1-51 1428:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 26 dep acids 1434:1438 arg1 1-51 1440:1443 1-51 1440:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 3 27 theme mammalian 401:409 arg1 species 411:417 mammalian species 401:417 mammalian species 401:417 Whereas Asn(92) is only conserved in mammalian species, Asn(211) is highly conserved in mammals, birds and fish. 25916169 7 28 theme ST3Gal-II 1052:1060 arg1 activity 1072:1079 ST3Gal-II enzymatic activity 1052:1079 ST3Gal-II enzymatic activity 1052:1079 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 6 29 theme pharmacological 735:749 arg1 mutagenesis 782:792 pharmacological, biochemical and site-directed mutagenesis 735:792 pharmacological, biochemical and site-directed mutagenesis 735:792 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 0 30 dep trafficking 87:97 arg1 the 69:71 the 69:71 the 69:71 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 4 31 theme human 621:625 arg1 ST3Gal-II 627:635 human ST3Gal-II 621:635 human ST3Gal-II 621:635 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 0 32 theme Critical 0:7 arg1 role 9:12 Critical role 0:12 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 0:142 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 4 33 theme present 481:487 arg1 study 489:493 The present study 477:493 The present study 477:493 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 6 34 from reticulum 949:957 arg1 exit 923:926 its exit 919:926 its exit from the endoplasmic reticulum and proper Golgi localization 919:987 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 10 35 theme endoplasmic 1694:1704 arg1 reticulum 1706:1714 the endoplasmic reticulum 1690:1714 the endoplasmic reticulum 1690:1714 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 8 36 theme acceptor 1223:1230 arg1 glycoprotein 1207:1218 glycoprotein 1207:1218 glycoprotein 1207:1218 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 36 theme acceptor 1223:1230 arg1 substrates 1232:1241 acceptor substrates 1223:1241 acceptor substrates 1223:1241 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 36 theme acceptor 1223:1230 arg1 glycolipid 1192:1201 glycolipid 1192:1201 glycolipid 1192:1201 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 1 37 theme main 195:198 arg1 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 37 theme main 195:198 arg1 sialyltransferase 210:226 the main mammalian sialyltransferase 191:226 the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain 191:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 2 38 contain contains 296:303 arg2 Asn 341:343 Asn 341:343 Asn(92) 341:347 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 38 contain contains 296:303 arg1 It 293:294 It 293:294 It 293:294 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 38 contain contains 296:303 arg2 sites 334:338 two putative N-glycosylation sites 305:338 two putative N-glycosylation sites (Asn(92) and Asn(211)) 305:361 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 38 contain contains 296:303 arg2 Asn 353:355 Asn 353:355 Asn(211) 353:360 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 8 39 from Asn 1274:1276 arg1 mutant 1264:1269 the single N-glycan mutant 1244:1269 the single N-glycan mutant at Asn(92) 1244:1280 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 9 40 theme endoplasmic 1493:1503 arg1 reticulum 1505:1513 the endoplasmic reticulum 1489:1513 the endoplasmic reticulum 1489:1513 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 6 41 gly N-glycosylated 832:845 arg2 211 854:856 211 854:856 211 854:856 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 6 41 gly N-glycosylated 832:845 arg2 Asn 850:852 Asn 850:852 Asn(211) 850:857 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 6 41 gly N-glycosylated 832:845 arg1 ST3Gal-II 812:820 ST3Gal-II 812:820 ST3Gal-II 812:820 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 1 42 theme mammalian 200:208 arg1 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 42 theme mammalian 200:208 arg1 sialyltransferase 210:226 the main mammalian sialyltransferase 191:226 the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain 191:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 10 43 theme complex 1777:1783 arg1 retention 1785:1793 Golgi complex retention 1771:1793 Golgi complex retention of the enzyme 1771:1807 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 4 44 theme intracellular 536:548 arg1 trafficking 550:560 intracellular trafficking 536:560 intracellular trafficking 536:560 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 8 45 contain had 1282:1284 arg1 mutant 1264:1269 the single N-glycan mutant 1244:1269 the single N-glycan mutant at Asn(92) 1244:1280 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 45 contain had 1282:1284 arg2 effect 1302:1307 only a moderate effect 1286:1307 only a moderate effect 1286:1307 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 2 46 theme N-glycosylation 318:332 arg1 sites 334:338 two putative N-glycosylation sites 305:338 two putative N-glycosylation sites (Asn(92) and Asn(211)) 305:361 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 46 theme N-glycosylation 318:332 arg1 Asn 353:355 Asn 353:355 Asn(211) 353:360 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 46 theme N-glycosylation 318:332 arg1 Asn 341:343 Asn 341:343 Asn(92) 341:347 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 4 47 from activity 573:580 arg1 ST3Gal-II 627:635 human ST3Gal-II 621:635 human ST3Gal-II 621:635 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 4 48 theme enzyme 566:571 arg1 activity 573:580 enzyme activity 566:580 enzyme activity 566:580 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 0 49 theme glycosylation 42:54 arg1 role 9:12 Critical role 0:12 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 0:142 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 8 50 theme single 1248:1253 arg1 mutant 1264:1269 the single N-glycan mutant 1244:1269 the single N-glycan mutant at Asn(92) 1244:1280 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 1 51 from biosynthesis 270:281 arg1 brain 286:290 brain 286:290 brain 286:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 10 52 theme Golgi 1771:1775 arg1 retention 1785:1793 Golgi complex retention 1771:1793 Golgi complex retention of the enzyme 1771:1807 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 9 53 theme transmembrane 1397:1409 arg1 region 1420:1425 the cytosolic, transmembrane and stem region 1382:1425 region 1420:1425 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 53 theme transmembrane 1397:1409 arg1 acids 1434:1438 amino acids 1-51 1428:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 1 54 theme responsible 228:238 arg1 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 54 theme responsible 228:238 arg1 sialyltransferase 210:226 the main mammalian sialyltransferase 191:226 the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain 191:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 0 55 theme conserved 32:40 arg1 glycosylation 42:54 evolutionarily conserved glycosylation 17:54 evolutionarily conserved glycosylation at Asn211 17:64 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 7 56 theme different 1031:1039 arg1 effects 1041:1047 different effects 1031:1047 different effects 1031:1047 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 4 57 dep occupancy 508:516 arg1 the 504:506 the 504:506 the 504:506 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 6 58 theme Golgi 970:974 arg1 localization 976:987 proper Golgi localization 963:987 proper Golgi localization 963:987 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 10 59 from reticulum 1706:1714 arg1 exit 1680:1683 proper exit 1673:1683 proper exit from the endoplasmic reticulum 1673:1714 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 9 60 theme cytosolic 1386:1394 arg1 region 1420:1425 the cytosolic, transmembrane and stem region 1382:1425 region 1420:1425 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 60 theme cytosolic 1386:1394 arg1 acids 1434:1438 amino acids 1-51 1428:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 7 61 theme N-glycosylation 1005:1019 arg1 sites 1021:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 8 62 theme position 1106:1113 arg1 211 1119:1121 211 1119:1121 211 1119:1121 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 62 theme position 1106:1113 arg1 Asn 1115:1117 position Asn 1106:1117 position Asn(211) 1106:1122 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 63 gly glycoprotein 1207:1218 arg1 glycoprotein 1207:1218 glycoprotein 1207:1218 glycoprotein 1207:1218 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 63 gly glycoprotein 1207:1218 arg1 substrates 1232:1241 acceptor substrates 1223:1241 acceptor substrates 1223:1241 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 8 63 gly glycoprotein 1207:1218 arg1 glycolipid 1192:1201 glycolipid 1192:1201 glycolipid 1192:1201 Whereas the N-glycan at position Asn(211) seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn(92) had only a moderate effect. 25916169 2 64 dep sites 334:338 arg1 sites 334:338 two putative N-glycosylation sites 305:338 two putative N-glycosylation sites (Asn(92) and Asn(211)) 305:361 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 64 dep sites 334:338 arg1 211 357:359 211 357:359 211 357:359 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 64 dep sites 334:338 arg1 92 345:346 92 345:346 92 345:346 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 64 dep sites 334:338 arg1 Asn 353:355 Asn 353:355 Asn(211) 353:360 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 2 64 dep sites 334:338 arg1 Asn 341:343 Asn 341:343 Asn(92) 341:347 It contains two putative N-glycosylation sites (Asn(92) and Asn(211)). 25916169 10 65 theme enzyme 1802:1807 arg1 retention 1785:1793 Golgi complex retention 1771:1793 Golgi complex retention of the enzyme 1771:1807 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 9 66 theme stem 1415:1418 arg1 region 1420:1425 the cytosolic, transmembrane and stem region 1382:1425 region 1420:1425 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 66 theme stem 1415:1418 arg1 acids 1434:1438 amino acids 1-51 1428:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 7 67 gly N-glycosylation 1005:1019 arg2 sites 1021:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 6 68 theme co-translational 873:888 arg1 modification 890:901 this co-translational modification 868:901 this co-translational modification 868:901 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 10 69 theme absolute 1746:1753 arg1 requirement 1755:1765 an absolute requirement 1743:1765 an absolute requirement for Golgi complex retention of the enzyme 1743:1807 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 9 70 theme amino 1428:1432 arg1 region 1420:1425 the cytosolic, transmembrane and stem region 1382:1425 region 1420:1425 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 9 70 theme amino 1428:1432 arg1 acids 1434:1438 amino acids 1-51 1428:1443 amino acids 1-51 1428:1443 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 10 71 theme C-terminal 1577:1586 arg1 ST3Gal-II 1598:1606 ST3Gal-II 1598:1606 ST3Gal-II 1598:1606 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 10 71 theme C-terminal 1577:1586 arg1 domain 1588:1593 the C-terminal domain 1573:1593 the C-terminal domain of ST3Gal-II 1573:1606 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 1 72 theme GD1a 244:247 arg1 biosynthesis 270:281 GD1a and GT1b ganglioside biosynthesis 244:281 GD1a and GT1b ganglioside biosynthesis in brain 244:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 7 73 theme individual 994:1003 arg1 sites 1021:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 0 74 gly glycosylation 42:54 arg2 Asn211 59:64 Asn211 59:64 Asn211 59:64 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 74 gly glycosylation 42:54 arg1 Asn211 59:64 Asn211 59:64 Asn211 59:64 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 0 74 gly glycosylation 42:54 arg1 sialyltransferase 115:131 sialyltransferase ST3Gal-II 115:141 sialyltransferase ST3Gal-II 115:141 Critical role of evolutionarily conserved glycosylation at Asn211 in the intracellular trafficking and activity of sialyltransferase ST3Gal-II. 25916169 7 75 contain had 1027:1029 arg2 effects 1041:1047 different effects 1031:1047 different effects 1031:1047 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 7 75 contain had 1027:1029 arg1 sites 1021:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites 990:1025 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 7 76 theme enzymatic 1062:1070 arg1 activity 1072:1079 ST3Gal-II enzymatic activity 1052:1079 ST3Gal-II enzymatic activity 1052:1079 The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. 25916169 9 77 theme Golgi 1539:1543 arg1 complex 1545:1551 the Golgi complex 1535:1551 the Golgi complex 1535:1551 Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. 25916169 10 78 theme optimum 1648:1654 arg1 conformation 1656:1667 an optimum conformation 1645:1667 an optimum conformation for proper exit from the endoplasmic reticulum 1645:1714 This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme. 25916169 4 79 theme potential 591:599 arg1 N-glycosylations 601:616 these potential N-glycosylations 585:616 these potential N-glycosylations 585:616 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 1 80 theme GT1b 253:256 arg1 biosynthesis 270:281 GD1a and GT1b ganglioside biosynthesis 244:281 GD1a and GT1b ganglioside biosynthesis in brain 244:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 4 81 from trafficking 550:560 arg1 ST3Gal-II 627:635 human ST3Gal-II 621:635 human ST3Gal-II 621:635 The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. 25916169 6 82 theme proper 963:968 arg1 localization 976:987 proper Golgi localization 963:987 proper Golgi localization 963:987 By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn(211) and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. 25916169 1 83 theme type II 157:163 arg1 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II 144:152 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 83 theme type II 157:163 arg1 protein 179:185 a type II transmembrane protein 155:185 a type II transmembrane protein 155:185 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 25916169 1 84 theme ganglioside 258:268 arg1 biosynthesis 270:281 GD1a and GT1b ganglioside biosynthesis 244:281 GD1a and GT1b ganglioside biosynthesis in brain 244:290 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. 24766575 4 0 gly glycopeptide 920:931 arg2 glycopeptide 920:931 glycopeptide profiling 920:941 glycopeptide profiling 920:941 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 1 1 theme analytical 192:201 arg1 challenge 203:211 a huge analytical challenge 185:211 a huge analytical challenge 185:211 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 3 2 theme spectra 664:670 arg1 thousands 636:644 thousands 636:644 thousands of N-glycopeptide spectra 636:670 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 4 3 theme similar 717:723 arg1 level 725:729 a similar level 715:729 a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins 715:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 3 4 gly N-glycopeptide 649:662 arg2 N-glycopeptide 649:662 N-glycopeptide spectra 649:670 N-glycopeptide spectra 649:670 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 1 5 theme N-glycopeptides 333:347 arg1 intricacy 320:328 an intricacy 317:328 an intricacy of N-glycopeptides 317:347 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 2 6 theme glycan 488:493 arg1 fragmentation 495:507 glycan fragmentation 488:507 glycan fragmentation in mass spectrometry 488:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 3 7 theme N-glycopeptide 649:662 arg1 spectra 664:670 N-glycopeptide spectra 649:670 N-glycopeptide spectra 649:670 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 4 8 from approach 794:801 arg1 proteins 817:824 individual proteins 806:824 individual proteins 806:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 1 9 theme protein 221:227 arg1 glycosylation 229:241 protein glycosylation 221:241 protein glycosylation 221:241 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 4 10 theme individual 806:815 arg1 proteins 817:824 individual proteins 806:824 individual proteins 806:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 0 11 theme Efficient 0:8 arg1 identification 36:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 4 12 theme site-specific 859:871 arg1 information 873:883 the unique in-depth site-specific information 839:883 the unique in-depth site-specific information that could only be studied through glycopeptide profiling 839:941 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 1 13 gly N-glycopeptides 333:347 arg2 N-glycopeptides 333:347 N-glycopeptides 333:347 N-glycopeptides 333:347 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 2 14 theme novel 375:379 arg1 approach 381:388 a novel approach 373:388 a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry 373:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 0 15 theme glycopeptide 23:34 arg1 identification 36:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 1 16 theme site-specific 129:141 arg1 N-glycosylation 143:157 site-specific N-glycosylation 129:157 site-specific N-glycosylation 129:157 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 0 17 theme accurate 14:21 arg1 identification 36:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 1 18 theme N-glycosylation 143:157 arg1 Study 120:124 Study 120:124 Study of site-specific N-glycosylation in complex sample 120:175 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 2 19 theme mass 512:515 arg1 spectrometry 517:528 mass spectrometry 512:528 mass spectrometry 512:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 2 20 theme N-glycopeptide 410:423 arg1 profiling 425:433 high-throughput N-glycopeptide profiling 394:433 high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry 394:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 4 21 theme conventional 771:782 arg1 approach 794:801 conventional glycomics approach 771:801 conventional glycomics approach on individual proteins 771:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 3 22 theme extensive 547:555 arg1 validation 557:566 an extensive validation 544:566 an extensive validation 544:566 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 4 23 theme unique 843:848 arg1 information 873:883 the unique in-depth site-specific information 839:883 the unique in-depth site-specific information that could only be studied through glycopeptide profiling 839:941 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 1 24 theme post-translational 270:287 arg1 modifications 289:301 post-translational modifications 270:301 post-translational modifications 270:301 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 0 25 theme high-throughput 64:78 arg1 analysis 110:117 high-throughput site-specific N-glycosylation analysis 64:117 high-throughput site-specific N-glycosylation analysis 64:117 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 2 26 gly N-glycopeptide 410:423 arg2 N-glycopeptide 410:423 high-throughput N-glycopeptide profiling 394:433 high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry 394:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 4 27 theme in-depth 850:857 arg1 information 873:883 the unique in-depth site-specific information 839:883 the unique in-depth site-specific information that could only be studied through glycopeptide profiling 839:941 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 0 28 gly glycopeptide 23:34 arg2 glycopeptide 23:34 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification 0:49 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 1 29 theme complex 162:168 arg1 sample 170:175 complex sample 162:175 complex sample 162:175 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 1 30 from modifications 289:301 arg1 diverse 259:265 diverse 259:265 diverse 259:265 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 1 31 from diverse 259:265 arg1 modifications 289:301 post-translational modifications 270:301 post-translational modifications 270:301 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 2 32 theme high-throughput 394:408 arg1 profiling 425:433 high-throughput N-glycopeptide profiling 394:433 high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry 394:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 3 33 theme high 677:680 arg1 confidence 682:691 high confidence 677:691 high confidence 677:691 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 4 34 theme glycopeptide 920:931 arg1 profiling 933:941 glycopeptide profiling 920:941 glycopeptide profiling 920:941 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 3 35 theme N-glycosylation 590:604 arg1 study 606:610 a high-throughput N-glycosylation study 572:610 a high-throughput N-glycosylation study 572:610 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 1 36 from Study 120:124 arg1 sample 170:175 complex sample 162:175 complex sample 162:175 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 3 37 theme high-throughput 574:588 arg1 study 606:610 a high-throughput N-glycosylation study 572:610 a high-throughput N-glycosylation study 572:610 We performed an extensive validation and a high-throughput N-glycosylation study on serum and identified thousands of N-glycopeptide spectra with high confidence. 24766575 0 38 theme N-glycosylation 94:108 arg1 analysis 110:117 high-throughput site-specific N-glycosylation analysis 64:117 high-throughput site-specific N-glycosylation analysis 64:117 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 2 39 from fragmentation 495:507 arg1 spectrometry 517:528 mass spectrometry 512:528 mass spectrometry 512:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 2 40 theme network-centric 446:460 arg1 algorithm 462:470 a network-centric algorithm 444:470 a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry 444:528 Here we have developed a novel approach for high-throughput N-glycopeptide profiling based on a network-centric algorithm for deciphering glycan fragmentation in mass spectrometry. 24766575 4 41 theme glycomics 784:792 arg1 approach 794:801 conventional glycomics approach 771:801 conventional glycomics approach on individual proteins 771:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 4 42 theme microheterogeneity 741:758 arg1 level 725:729 a similar level 715:729 a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins 715:824 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 24766575 0 43 theme site-specific 80:92 arg1 analysis 110:117 high-throughput site-specific N-glycosylation analysis 64:117 high-throughput site-specific N-glycosylation analysis 64:117 Efficient and accurate glycopeptide identification pipeline for high-throughput site-specific N-glycosylation analysis. 24766575 1 44 theme huge 187:190 arg1 challenge 203:211 a huge analytical challenge 185:211 a huge analytical challenge 185:211 Study of site-specific N-glycosylation in complex sample remains a huge analytical challenge because protein glycosylation is structurally diverse in post-translational modifications, resulting in an intricacy of N-glycopeptides. 24766575 4 45 theme glycan 734:739 arg1 microheterogeneity 741:758 glycan microheterogeneity 734:758 glycan microheterogeneity 734:758 The results revealed a similar level of glycan microheterogeneity to that of conventional glycomics approach on individual proteins and provided the unique in-depth site-specific information that could only be studied through glycopeptide profiling. 26956484 4 0 theme carbohydrate-binding 498:517 arg1 proteins 519:526 carbohydrate-binding proteins 498:526 carbohydrate-binding proteins 498:526 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 4 0 theme carbohydrate-binding 498:517 arg1 Lectins 486:492 Lectins 486:492 Lectins 486:492 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 2 1 gly N-glycosylated 275:288 arg1 subunits 290:297 N-glycosylated subunits 275:297 N-glycosylated subunits 275:297 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 7 2 theme Orai1 1057:1061 arg1 structure 1044:1052 the glycan structure 1033:1052 the glycan structure of Orai1 1033:1061 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 7 3 link α-2,6-linked 1004:1015 arg1 acids 1024:1028 α-2,6-linked sialic acids 1004:1028 α-2,6-linked sialic acids 1004:1028 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 10 4 theme Orai1 1452:1456 arg1 state 1443:1447 the glycosylation state 1425:1447 the glycosylation state of Orai1 1425:1456 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 8 5 theme mast 1138:1141 arg1 cells 1143:1147 human mast cells 1132:1147 human mast cells 1132:1147 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 3 6 theme Ca 471:472 arg1 influx 478:483 Ca(2+) influx 471:483 Ca(2+) influx 471:483 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 9 7 theme Orai1 1339:1343 arg1 interaction 1324:1334 the interaction 1320:1334 the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE 1320:1410 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 26956484 6 8 theme imaging 796:802 arg1 experiments 804:814 Ca(2+) imaging experiments 789:814 Ca(2+) imaging experiments 789:814 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 9 9 theme specific 1350:1357 arg1 Siglecs 1376:1382 Siglecs 1376:1382 Siglecs 1376:1382 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 26956484 9 9 theme specific 1350:1357 arg1 lectins 1359:1365 specific lectins 1350:1365 specific lectins 1350:1365 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 26956484 6 10 theme Orai1 901:905 arg1 Orai1 901:905 Orai1 901:905 Orai1 901:905 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 10 theme Orai1 901:905 arg1 site 893:896 the only glycosylation site 870:896 the only glycosylation site of Orai1 (Orai1N223A) 870:918 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 10 11 theme SOCE-mediated 1469:1481 arg1 2+ 1486:1487 2+ 1486:1487 2+ 1486:1487 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 10 11 theme SOCE-mediated 1469:1481 arg1 Ca 1483:1484 SOCE-mediated Ca 1469:1484 SOCE-mediated Ca(2+) signaling 1469:1498 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 7 12 theme α-2,6-linked 1004:1015 arg1 acids 1024:1028 α-2,6-linked sialic acids 1004:1028 α-2,6-linked sialic acids 1004:1028 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 6 13 theme site 893:896 arg1 mutation 858:865 mutation 858:865 mutation of the only glycosylation site of Orai1 (Orai1N223A) 858:918 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 7 14 from entry 1104:1108 arg1 cells 1122:1126 Jurkat T cells 1113:1126 Jurkat T cells 1113:1126 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 4 15 theme acid-binding 564:575 arg1 lectins 577:583 sialic acid-binding lectins 557:583 sialic acid-binding lectins with immunoglobulin-like repeats 557:616 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 7 16 theme ST6GAL1 988:994 arg1 Knockdown 953:961 Knockdown 953:961 Knockdown of the sialyltransferase ST6GAL1 953:994 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 6 17 theme only 874:877 arg1 Orai1 901:905 Orai1 901:905 Orai1 901:905 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 17 theme only 874:877 arg1 site 893:896 the only glycosylation site 870:896 the only glycosylation site of Orai1 (Orai1N223A) 870:918 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 10 18 located observed 1565:1572 arg1 cancer 1596:1601 cancer 1596:1601 cancer 1596:1601 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 10 18 located observed 1565:1572 arg1 disease 1584:1590 immune disease 1577:1590 immune disease 1577:1590 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 10 18 located observed 1565:1572 arg2 signaling 1555:1563 pathophysiological Ca(2+) signaling 1529:1563 pathophysiological Ca(2+) signaling observed in immune disease and cancer 1529:1601 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 2 19 theme store-operated 223:236 arg1 entry 245:249 store-operated Ca(2+) entry 223:249 store-operated Ca(2+) entry (SOCE) 223:256 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 2 19 theme store-operated 223:236 arg1 SOCE 252:255 SOCE 252:255 SOCE 252:255 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 5 20 theme blot 633:636 arg1 analysis 638:645 Western blot analysis 625:645 Western blot analysis 625:645 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 6 21 theme T 944:944 arg1 cells 946:950 Jurkat T cells 937:950 Jurkat T cells 937:950 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 10 22 gly glycosylation 1429:1441 arg1 Orai1 1452:1456 Orai1 1452:1456 Orai1 1452:1456 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 7 23 theme Ca 1097:1098 arg1 entry 1104:1108 increased Ca(2+) entry 1087:1108 increased Ca(2+) entry in Jurkat T cells 1087:1126 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 8 24 theme other 1216:1220 arg1 proteins 1222:1229 other proteins 1216:1229 other proteins 1216:1229 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 8 24 theme other 1216:1220 arg1 Orai1 1205:1209 Orai1 1205:1209 Orai1 (and other proteins) 1205:1230 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 1 25 gly N-glycosylation 79:93 arg1 protein 128:134 protein function 128:143 protein function 128:143 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 1 25 gly N-glycosylation 79:93 arg1 proteins 111:118 cell surface proteins 98:118 cell surface proteins 98:118 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 7 26 theme Jurkat 1113:1118 arg1 cells 1122:1126 Jurkat T cells 1113:1126 Jurkat T cells 1113:1126 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 3 27 dep molecules 363:371 arg1 STIM1 373:377 STIM1 373:377 STIM1 373:377 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 27 dep molecules 363:371 arg1 molecules 363:371 stromal interaction molecules STIM1 or STIM2 343:386 stromal interaction molecules STIM1 or STIM2 343:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 27 dep molecules 363:371 arg1 STIM2 382:386 STIM2 382:386 STIM2 382:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 28 theme Ca 424:425 arg1 channels 431:438 Orai Ca(2+) channels 419:438 Orai Ca(2+) channels in the plasma membrane 419:461 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 5 29 theme human 694:698 arg1 cells 700:704 various primary human cells 678:704 various primary human cells 678:704 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 1 30 with stability 146:154 arg1 proteins 184:191 other proteins 178:191 other proteins 178:191 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 3 31 theme stromal 343:349 arg1 STIM1 373:377 STIM1 373:377 STIM1 373:377 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 31 theme stromal 343:349 arg1 proteins 333:340 Ca(2+) sensor proteins 319:340 Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) 319:387 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 31 theme stromal 343:349 arg1 molecules 363:371 stromal interaction molecules STIM1 or STIM2 343:386 stromal interaction molecules STIM1 or STIM2 343:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 31 theme stromal 343:349 arg1 STIM2 382:386 STIM2 382:386 STIM2 382:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 5 32 theme lectin-binding 651:664 arg1 assays 666:671 lectin-binding assays 651:671 lectin-binding assays 651:671 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 1 33 with function 136:143 arg1 proteins 184:191 other proteins 178:191 other proteins 178:191 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 1 34 theme surface 103:109 arg1 proteins 111:118 cell surface proteins 98:118 cell surface proteins 98:118 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 3 35 theme Orai 419:422 arg1 2+ 427:428 2+ 427:428 2+ 427:428 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 35 theme Orai 419:422 arg1 Ca 424:425 Orai Ca 419:425 Orai Ca(2+) channels in the plasma membrane 419:461 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 5 36 theme Orai1 760:764 arg1 glycosylation 743:755 glycosylation 743:755 glycosylation of Orai1 743:764 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 5 37 gly glycosylation 743:755 arg1 Orai1 760:764 Orai1 760:764 Orai1 760:764 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 10 38 theme pathophysiological 1529:1546 arg1 signaling 1555:1563 pathophysiological Ca(2+) signaling 1529:1563 pathophysiological Ca(2+) signaling observed in immune disease and cancer 1529:1601 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 0 39 theme Orai1 36:40 arg1 glycosylation 19:31 Cell type-specific glycosylation 0:31 Cell type-specific glycosylation of Orai1 0:40 Cell type-specific glycosylation of Orai1 modulates store-operated Ca2+ entry. 26956484 4 40 with lectins 577:583 arg1 repeats 610:616 immunoglobulin-like repeats 590:616 immunoglobulin-like repeats 590:616 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 9 41 with interaction 1324:1334 arg1 Siglecs 1376:1382 Siglecs 1376:1382 Siglecs 1376:1382 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 26956484 9 41 with interaction 1324:1334 arg1 lectins 1359:1365 specific lectins 1350:1365 specific lectins 1350:1365 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 26956484 0 42 theme store-operated 52:65 arg1 entry 72:76 store-operated Ca2+ entry 52:76 store-operated Ca2+ entry 52:76 Cell type-specific glycosylation of Orai1 modulates store-operated Ca2+ entry. 26956484 10 43 theme glycosylation 1429:1441 arg1 state 1443:1447 the glycosylation state 1425:1447 the glycosylation state of Orai1 1425:1456 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 3 44 theme plasma 447:452 arg1 membrane 454:461 the plasma membrane 443:461 the plasma membrane 443:461 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 4 45 theme lectins 577:583 arg1 proteins 519:526 carbohydrate-binding proteins 498:526 carbohydrate-binding proteins 498:526 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 4 45 theme lectins 577:583 arg1 Siglecs 533:539 Siglecs 533:539 Siglecs 533:539 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 4 45 theme lectins 577:583 arg1 family 547:552 a family 545:552 a family of sialic acid-binding lectins with immunoglobulin-like repeats 545:616 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 4 45 theme lectins 577:583 arg1 Lectins 486:492 Lectins 486:492 Lectins 486:492 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 7 46 theme T 1120:1120 arg1 cells 1122:1126 Jurkat T cells 1113:1126 Jurkat T cells 1113:1126 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 10 47 theme immune 1577:1582 arg1 disease 1584:1590 immune disease 1577:1590 immune disease 1577:1590 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 7 48 theme glycan 1037:1042 arg1 structure 1044:1052 the glycan structure 1033:1052 the glycan structure of Orai1 1033:1061 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 6 49 gly glycosylation 879:891 arg1 Orai1N223A 908:917 Orai1N223A 908:917 Orai1N223A 908:917 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 49 gly glycosylation 879:891 arg2 Orai1 901:905 Orai1 901:905 Orai1 901:905 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 49 gly glycosylation 879:891 arg1 Orai1 901:905 Orai1 901:905 Orai1 901:905 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 49 gly glycosylation 879:891 arg2 site 893:896 the only glycosylation site 870:896 the only glycosylation site of Orai1 (Orai1N223A) 870:918 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 50 theme patch-clamp 820:830 arg1 experiments 832:842 patch-clamp experiments 820:842 patch-clamp experiments 820:842 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 1 51 theme other 178:182 arg1 proteins 184:191 other proteins 178:191 other proteins 178:191 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 8 52 theme human 1132:1136 arg1 cells 1143:1147 human mast cells 1132:1147 human mast cells 1132:1147 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 5 53 theme cell 717:720 arg1 lines 722:726 cancer cell lines 710:726 cancer cell lines 710:726 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 7 54 theme sialic 1017:1022 arg1 acids 1024:1028 α-2,6-linked sialic acids 1004:1028 α-2,6-linked sialic acids 1004:1028 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 4 55 theme immunoglobulin-like 590:608 arg1 repeats 610:616 immunoglobulin-like repeats 590:616 immunoglobulin-like repeats 590:616 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 10 56 theme Ca 1483:1484 arg1 signaling 1490:1498 SOCE-mediated Ca(2+) signaling 1469:1498 SOCE-mediated Ca(2+) signaling 1469:1498 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 3 57 from channels 431:438 arg1 membrane 454:461 the plasma membrane 443:461 the plasma membrane 443:461 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 2 58 theme N-glycosylated 275:288 arg1 subunits 290:297 N-glycosylated subunits 275:297 N-glycosylated subunits 275:297 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 7 59 theme sialyltransferase 970:986 arg1 ST6GAL1 988:994 the sialyltransferase ST6GAL1 966:994 the sialyltransferase ST6GAL1 966:994 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 6 60 theme glycosylation 879:891 arg1 Orai1 901:905 Orai1 901:905 Orai1 901:905 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 6 60 theme glycosylation 879:891 arg1 site 893:896 the only glycosylation site 870:896 the only glycosylation site of Orai1 (Orai1N223A) 870:918 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 4 61 theme sialic 557:562 arg1 lectins 577:583 sialic acid-binding lectins 557:583 sialic acid-binding lectins with immunoglobulin-like repeats 557:616 Lectins are carbohydrate-binding proteins, and Siglecs are a family of sialic acid-binding lectins with immunoglobulin-like repeats. 26956484 2 62 theme Ca 238:239 arg1 entry 245:249 store-operated Ca(2+) entry 223:249 store-operated Ca(2+) entry (SOCE) 223:256 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 2 62 theme Ca 238:239 arg1 SOCE 252:255 SOCE 252:255 SOCE 252:255 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 1 63 theme protein 128:134 arg1 function 136:143 protein function 128:143 protein function 128:143 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 5 64 theme Western 625:631 arg1 analysis 638:645 Western blot analysis 625:645 Western blot analysis 625:645 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 3 65 from activation 305:314 arg1 reticulum 408:416 the endoplasmic reticulum 392:416 the endoplasmic reticulum 392:416 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 6 66 theme Jurkat 937:942 arg1 cells 946:950 Jurkat T cells 937:950 Jurkat T cells 937:950 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 3 67 theme Ca 319:320 arg1 proteins 333:340 Ca(2+) sensor proteins 319:340 Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) 319:387 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 67 theme Ca 319:320 arg1 molecules 363:371 stromal interaction molecules STIM1 or STIM2 343:386 stromal interaction molecules STIM1 or STIM2 343:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 8 68 theme Orai1 1205:1209 arg1 N-glycan 1193:1200 the N-glycan 1189:1200 the N-glycan of Orai1 (and other proteins) 1189:1230 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 5 69 theme various 678:684 arg1 cells 700:704 various primary human cells 678:704 various primary human cells 678:704 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 5 70 theme cancer 710:715 arg1 lines 722:726 cancer cell lines 710:726 cancer cell lines 710:726 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 1 71 with interaction 161:171 arg1 proteins 184:191 other proteins 178:191 other proteins 178:191 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 3 72 theme sensor 326:331 arg1 proteins 333:340 Ca(2+) sensor proteins 319:340 Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) 319:387 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 72 theme sensor 326:331 arg1 molecules 363:371 stromal interaction molecules STIM1 or STIM2 343:386 stromal interaction molecules STIM1 or STIM2 343:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 5 73 theme primary 686:692 arg1 cells 700:704 various primary human cells 678:704 various primary human cells 678:704 Using Western blot analysis and lectin-binding assays from various primary human cells and cancer cell lines, we found that glycosylation of Orai1 is cell type-specific. 26956484 10 74 dep influences 1458:1467 arg1 contribute 1515:1524 contribute 1515:1524 may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer 1511:1601 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 3 75 theme endoplasmic 396:406 arg1 reticulum 408:416 the endoplasmic reticulum 392:416 the endoplasmic reticulum 392:416 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 0 76 theme type-specific 5:17 arg1 glycosylation 19:31 Cell type-specific glycosylation 0:31 Cell type-specific glycosylation of Orai1 0:40 Cell type-specific glycosylation of Orai1 modulates store-operated Ca2+ entry. 26956484 1 77 theme cell 98:101 arg1 proteins 111:118 cell surface proteins 98:118 cell surface proteins 98:118 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 3 78 theme interaction 351:361 arg1 STIM1 373:377 STIM1 373:377 STIM1 373:377 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 78 theme interaction 351:361 arg1 proteins 333:340 Ca(2+) sensor proteins 319:340 Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) 319:387 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 78 theme interaction 351:361 arg1 molecules 363:371 stromal interaction molecules STIM1 or STIM2 343:386 stromal interaction molecules STIM1 or STIM2 343:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 3 78 theme interaction 351:361 arg1 STIM2 382:386 STIM2 382:386 STIM2 382:386 Upon activation by Ca(2+) sensor proteins (stromal interaction molecules STIM1 or STIM2) in the endoplasmic reticulum, Orai Ca(2+) channels in the plasma membrane mediate Ca(2+) influx. 26956484 7 79 theme increased 1087:1095 arg1 entry 1104:1108 increased Ca(2+) entry 1087:1108 increased Ca(2+) entry in Jurkat T cells 1087:1126 Knockdown of the sialyltransferase ST6GAL1 reduced α-2,6-linked sialic acids in the glycan structure of Orai1 and was associated with increased Ca(2+) entry in Jurkat T cells. 26956484 1 80 theme proteins 111:118 arg1 N-glycosylation 79:93 N-glycosylation 79:93 N-glycosylation of cell surface proteins 79:118 N-glycosylation of cell surface proteins affects protein function, stability, and interaction with other proteins. 26956484 2 81 theme Orai 194:197 arg1 channels 199:206 Orai channels 194:206 Orai channels 194:206 Orai channels, which mediate store-operated Ca(2+) entry (SOCE), are composed of N-glycosylated subunits. 26956484 10 82 theme Ca 1548:1549 arg1 signaling 1555:1563 pathophysiological Ca(2+) signaling 1529:1563 pathophysiological Ca(2+) signaling observed in immune disease and cancer 1529:1601 In summary, the glycosylation state of Orai1 influences SOCE-mediated Ca(2+) signaling and, thus, may contribute to pathophysiological Ca(2+) signaling observed in immune disease and cancer. 26956484 8 83 theme sulfation 1171:1179 arg1 inhibition 1150:1159 inhibition 1150:1159 inhibition of sialyl sulfation 1150:1179 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 0 84 theme Ca2+ 67:70 arg1 entry 72:76 store-operated Ca2+ entry 52:76 store-operated Ca2+ entry 52:76 Cell type-specific glycosylation of Orai1 modulates store-operated Ca2+ entry. 26956484 6 85 theme Ca 789:790 arg1 experiments 804:814 Ca(2+) imaging experiments 789:814 Ca(2+) imaging experiments 789:814 Ca(2+) imaging experiments and patch-clamp experiments revealed that mutation of the only glycosylation site of Orai1 (Orai1N223A) enhanced SOCE in Jurkat T cells. 26956484 0 86 gly glycosylation 19:31 arg1 Orai1 36:40 Orai1 36:40 Orai1 36:40 Cell type-specific glycosylation of Orai1 modulates store-operated Ca2+ entry. 26956484 8 87 theme sialyl 1164:1169 arg1 sulfation 1171:1179 sialyl sulfation 1164:1179 sialyl sulfation 1164:1179 In human mast cells, inhibition of sialyl sulfation altered the N-glycan of Orai1 (and other proteins) and increased SOCE. 26956484 9 88 theme type-specific 1281:1293 arg1 glycosylation 1295:1307 cell type-specific glycosylation 1276:1307 cell type-specific glycosylation 1276:1307 These data suggest that cell type-specific glycosylation influences the interaction of Orai1 with specific lectins, such as Siglecs, which then attenuates SOCE. 24511137 5 0 attach attached 1136:1143 arg1 regions 1180:1186 the Fc regions 1173:1186 the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1173:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 5 0 attach attached 1136:1143 arg1 sites 1164:1168 three different sites 1148:1168 three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1148:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 5 0 attach attached 1136:1143 arg2 glycans 1128:1134 N-linked glycans 1119:1134 N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1119:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 2 1 theme class-switch 375:386 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 1 2 theme aberrations 264:274 arg1 contribution 222:233 the pathogenic contribution 207:233 the pathogenic contribution of the underlying structural aberrations 207:274 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 6 3 theme O-linked 1288:1295 arg1 glycans 1297:1303 O-linked glycans 1288:1303 O-linked glycans attached to the hinge region 1288:1332 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 3 4 theme hinge 590:594 arg1 region 596:601 the hinge region 586:601 the hinge region 586:601 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 5 from role 1280:1283 arg1 development 1341:1351 the development 1337:1351 the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1337:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 0 6 link O-linked 0:7 arg1 glycosylation 9:21 O-linked glycosylation 0:21 O-linked glycosylation 0:21 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 6 7 link O-linked 1288:1295 arg1 glycans 1297:1303 O-linked glycans 1288:1303 O-linked glycans attached to the hinge region 1288:1332 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 5 8 theme 6-19 1227:1230 arg1 IgAs 1232:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 4 9 theme implanted 869:877 arg1 cells 879:883 implanted cells 869:883 implanted cells 869:883 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 10 link N-linked 747:754 arg1 glycans 756:762 O- and N-linked glycans 740:762 glycans 756:762 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 11 theme glycans 756:762 arg1 contributions 723:735 the respective contributions 708:735 the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice 708:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 4 12 theme 6-19 968:971 arg1 glycovariants 977:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 5 13 theme IgAs 1232:1235 arg1 regions 1180:1186 the Fc regions 1173:1186 the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1173:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 1 14 theme pathogenic 211:220 arg1 contribution 222:233 the pathogenic contribution 207:233 the pathogenic contribution of the underlying structural aberrations 207:274 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 15 theme glomerular 918:927 arg1 lesions 929:935 glomerular lesions 918:935 glomerular lesions 918:935 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 16 theme N-linked 625:632 arg1 glycans 634:640 N-linked glycans 625:640 N-linked glycans 625:640 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 17 theme factor 822:827 arg1 potential 785:793 the nephritogenic potential 767:793 the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice 767:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 1 18 theme hinge 144:148 arg1 region 150:155 the IgA1 hinge region 135:155 the IgA1 hinge region 135:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 19 gly O-glycosylated 953:966 arg1 glycovariants 977:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 4 20 theme IgA 1001:1003 arg1 mutant 1011:1016 a 6-19 IgA hinge mutant 994:1016 a 6-19 IgA hinge mutant lacking O-linked glycans 994:1041 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 21 from structures 611:620 arg1 domain 653:658 the CH1 domain 645:658 the CH1 domain 645:658 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 21 from structures 611:620 arg1 region 596:601 the hinge region 586:601 the hinge region 586:601 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 22 theme IgA 807:809 arg1 factor 822:827 the 6-19 IgA rheumatoid factor 798:827 the 6-19 IgA rheumatoid factor in mice 798:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 0 23 theme rheumatoid 69:78 arg1 factor 80:85 IgA rheumatoid factor 65:85 IgA rheumatoid factor 65:85 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 3 24 theme CH1 649:651 arg1 domain 653:658 the CH1 domain 645:658 the CH1 domain 645:658 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 25 from potential 785:793 arg1 mice 832:835 mice 832:835 mice 832:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 26 theme 6-19 1391:1394 arg1 factor 1411:1416 6-19 IgA rheumatoid factor 1391:1416 6-19 IgA rheumatoid factor in mice 1391:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 5 27 theme different 1154:1162 arg1 regions 1180:1186 the Fc regions 1173:1186 the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1173:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 5 27 theme different 1154:1162 arg1 sites 1164:1168 three different sites 1148:1168 three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1148:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 1 28 gly glycosylation 98:110 arg1 region 150:155 the IgA1 hinge region 135:155 the IgA1 hinge region 135:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 1 28 gly glycosylation 98:110 arg1 glycans 124:130 O-linked glycans 115:130 O-linked glycans 115:130 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 6 29 theme nephropathy-like 1360:1375 arg1 GN 1377:1378 IgA nephropathy-like GN 1356:1378 IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1356:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 5 30 theme glycans 1128:1134 arg1 structure 1106:1114 the structure 1102:1114 the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1102:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 1 31 theme Deficient 88:96 arg1 glycosylation 98:110 Deficient glycosylation 88:110 Deficient glycosylation of O-linked glycans in the IgA1 hinge region 88:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 32 link O-linked 1026:1033 arg1 glycans 1035:1041 O-linked glycans 1026:1041 O-linked glycans 1026:1041 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 2 33 theme rheumatoid 416:425 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 0 34 theme O-linked 0:7 arg1 glycosylation 9:21 O-linked glycosylation 0:21 O-linked glycosylation 0:21 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 2 35 theme rheumatoid 464:473 arg1 factor 475:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 3 36 from mice 832:835 arg1 potential 785:793 the nephritogenic potential 767:793 the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice 767:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 2 37 theme IgA 449:451 arg1 factor 475:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 1 38 theme glycans 124:130 arg1 glycosylation 98:110 Deficient glycosylation 88:110 Deficient glycosylation of O-linked glycans in the IgA1 hinge region 88:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 39 theme O-linked 1026:1033 arg1 glycans 1035:1041 O-linked glycans 1026:1041 O-linked glycans 1026:1041 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 0 40 theme nephritogenic 38:50 arg1 potential 52:60 the nephritogenic potential 34:60 the nephritogenic potential of IgA rheumatoid factor 34:85 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 2 41 theme glomerular 491:500 arg1 lesions 502:508 glomerular lesions 491:508 glomerular lesions resembling IgA nephropathy 491:535 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 3 42 theme 6-19 670:673 arg1 IgA 675:677 6-19 IgA 670:677 6-19 IgA 670:677 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 4 43 theme 6-19 848:851 arg1 IgA 853:855 Wild-type 6-19 IgA 838:855 Wild-type 6-19 IgA secreted by implanted cells 838:883 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 5 44 theme Fc 1177:1178 arg1 regions 1180:1186 the Fc regions 1173:1186 the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1173:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 3 45 from factor 822:827 arg1 mice 832:835 mice 832:835 mice 832:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 1 46 link O-linked 115:122 arg1 glycans 124:130 O-linked glycans 115:130 O-linked glycans 115:130 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 2 47 theme IgA 401:403 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 3 48 theme O- 740:741 arg1 glycans 756:762 O- and N-linked glycans 740:762 glycans 756:762 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 2 49 theme variant 388:394 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 1 50 theme structural 253:262 arg1 aberrations 264:274 the underlying structural aberrations 238:274 the underlying structural aberrations 238:274 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 6 51 theme glycans 1297:1303 arg1 role 1280:1283 a critical role 1269:1283 a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1269:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 3 52 from levels 550:555 arg1 domain 653:658 the CH1 domain 645:658 the CH1 domain 645:658 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 52 from levels 550:555 arg1 region 596:601 the hinge region 586:601 the hinge region 586:601 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 4 53 theme significant 893:903 arg1 formation 905:913 significant formation 893:913 significant formation of glomerular lesions 893:935 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 54 theme N-linked 747:754 arg1 glycans 756:762 O- and N-linked glycans 740:762 glycans 756:762 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 55 theme glycosylation 569:581 arg1 levels 550:555 the levels 546:555 the levels of O-linked glycosylation in the hinge region 546:601 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 55 theme glycosylation 569:581 arg1 structures 611:620 the structures 607:620 the structures of N-linked glycans in the CH1 domain 607:658 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 1 56 theme IgA 176:178 arg1 nephropathy 180:190 IgA nephropathy 176:190 IgA nephropathy in humans 176:200 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 57 theme hinge 1005:1009 arg1 mutant 1011:1016 a 6-19 IgA hinge mutant 994:1016 a 6-19 IgA hinge mutant lacking O-linked glycans 994:1041 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 6 58 theme critical 1271:1278 arg1 role 1280:1283 a critical role 1269:1283 a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1269:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 6 59 from factor 1411:1416 arg1 mice 1421:1424 mice 1421:1424 mice 1421:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 6 60 attach attached 1305:1312 arg1 region 1327:1332 the hinge region 1317:1332 the hinge region 1317:1332 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 6 60 attach attached 1305:1312 arg2 glycans 1297:1303 O-linked glycans 1288:1303 O-linked glycans attached to the hinge region 1288:1332 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 3 61 theme nephritogenic 771:783 arg1 potential 785:793 the nephritogenic potential 767:793 the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice 767:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 62 theme IgA 1356:1358 arg1 GN 1377:1378 IgA nephropathy-like GN 1356:1378 IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1356:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 3 63 theme glycans 634:640 arg1 levels 550:555 the levels 546:555 the levels of O-linked glycosylation in the hinge region 546:601 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 63 theme glycans 634:640 arg1 structures 611:620 the structures 607:620 the structures of N-linked glycans in the CH1 domain 607:658 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 64 theme rheumatoid 1400:1409 arg1 factor 1411:1416 6-19 IgA rheumatoid factor 1391:1416 6-19 IgA rheumatoid factor in mice 1391:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 4 65 theme O-glycosylated 953:966 arg1 glycovariants 977:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 66 theme 6-19 802:805 arg1 factor 822:827 the 6-19 IgA rheumatoid factor 798:827 the 6-19 IgA rheumatoid factor in mice 798:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 67 theme hinge 1321:1325 arg1 region 1327:1332 the hinge region 1317:1332 the hinge region 1317:1332 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 1 68 theme IgA1 139:142 arg1 region 150:155 the IgA1 hinge region 135:155 the IgA1 hinge region 135:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 4 69 theme lesions 929:935 arg1 formation 905:913 significant formation 893:913 significant formation of glomerular lesions 893:935 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 0 70 theme factor 80:85 arg1 potential 52:60 the nephritogenic potential 34:60 the nephritogenic potential of IgA rheumatoid factor 34:85 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 5 71 theme apparent 1076:1083 arg1 heterogeneity 1085:1097 no apparent heterogeneity 1073:1097 no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1073:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 4 72 theme 6-19 996:999 arg1 mutant 1011:1016 a 6-19 IgA hinge mutant 994:1016 a 6-19 IgA hinge mutant lacking O-linked glycans 994:1041 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 3 73 theme rheumatoid 811:820 arg1 factor 822:827 the 6-19 IgA rheumatoid factor 798:827 the 6-19 IgA rheumatoid factor in mice 798:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 6 74 theme IgA 1396:1398 arg1 factor 1411:1416 6-19 IgA rheumatoid factor 1391:1416 6-19 IgA rheumatoid factor in mice 1391:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 1 75 from nephropathy 180:190 arg1 humans 195:200 humans 195:200 humans 195:200 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 2 76 theme IgA 521:523 arg1 nephropathy 525:535 IgA nephropathy 521:535 IgA nephropathy 521:535 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 4 77 theme IgA 973:975 arg1 glycovariants 977:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 poorly O-glycosylated 6-19 IgA glycovariants 946:989 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 5 78 from heterogeneity 1085:1097 arg1 structure 1106:1114 the structure 1102:1114 the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1102:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 5 79 theme N-linked 1119:1126 arg1 glycans 1128:1134 N-linked glycans 1119:1134 N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1119:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 6 80 theme GN 1377:1378 arg1 development 1341:1351 the development 1337:1351 the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice 1337:1424 Collectively, our data suggest a critical role of O-linked glycans attached to the hinge region in the development of IgA nephropathy-like GN induced by 6-19 IgA rheumatoid factor in mice. 24511137 5 81 link N-linked 1119:1126 arg1 glycans 1128:1134 N-linked glycans 1119:1134 N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1119:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 2 82 theme anti-IgG2a 405:414 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 5 83 theme nephritogenic 1191:1203 arg1 IgAs 1232:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 1 84 theme O-linked 115:122 arg1 glycans 124:130 O-linked glycans 115:130 O-linked glycans 115:130 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 2 85 theme anti-IgG2a 453:462 arg1 factor 475:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 2 86 theme 46-42 443:447 arg1 factor 475:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 46-42 IgA anti-IgG2a rheumatoid factor 443:480 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 5 87 theme non-nephritogenic 1209:1225 arg1 IgAs 1232:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 nephritogenic and non-nephritogenic 6-19 IgAs 1191:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 3 88 link N-linked 625:632 arg1 glycans 634:640 N-linked glycans 625:640 N-linked glycans 625:640 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 0 89 theme IgA 65:67 arg1 factor 80:85 IgA rheumatoid factor 65:85 IgA rheumatoid factor 65:85 O-linked glycosylation determines the nephritogenic potential of IgA rheumatoid factor. 24511137 3 90 link O-linked 560:567 arg1 glycosylation 569:581 O-linked glycosylation 560:581 O-linked glycosylation 560:581 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 4 91 theme Wild-type 838:846 arg1 IgA 853:855 Wild-type 6-19 IgA 838:855 Wild-type 6-19 IgA secreted by implanted cells 838:883 Wild-type 6-19 IgA secreted by implanted cells induced significant formation of glomerular lesions, whereas poorly O-glycosylated 6-19 IgA glycovariants or a 6-19 IgA hinge mutant lacking O-linked glycans did not. 24511137 5 92 theme regions 1180:1186 arg1 regions 1180:1186 the Fc regions 1173:1186 the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1173:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 5 92 theme regions 1180:1186 arg1 sites 1164:1168 three different sites 1148:1168 three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs 1148:1235 However, we observed no apparent heterogeneity in the structure of N-linked glycans attached to three different sites of the Fc regions of nephritogenic and non-nephritogenic 6-19 IgAs. 24511137 2 93 theme 6-19 396:399 arg1 factor 427:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor 371:432 We previously showed that mice implanted with cells secreting the class-switch variant 6-19 IgA anti-IgG2a rheumatoid factor, but not 46-42 IgA anti-IgG2a rheumatoid factor, develop glomerular lesions resembling IgA nephropathy. 24511137 1 94 from glycosylation 98:110 arg1 region 150:155 the IgA1 hinge region 135:155 the IgA1 hinge region 135:155 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 24511137 3 95 theme respective 712:721 arg1 contributions 723:735 the respective contributions 708:735 the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice 708:835 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 3 96 theme O-linked 560:567 arg1 glycosylation 569:581 O-linked glycosylation 560:581 O-linked glycosylation 560:581 Because the levels of O-linked glycosylation in the hinge region and the structures of N-linked glycans in the CH1 domain differ in 6-19 IgA and 46-42 IgA, we determined the respective contributions of O- and N-linked glycans to the nephritogenic potential of the 6-19 IgA rheumatoid factor in mice. 24511137 1 97 theme underlying 242:251 arg1 aberrations 264:274 the underlying structural aberrations 238:274 the underlying structural aberrations 238:274 Deficient glycosylation of O-linked glycans in the IgA1 hinge region is associated with IgA nephropathy in humans, but the pathogenic contribution of the underlying structural aberrations remains incompletely understood. 28958711 7 0 theme O-glycans 1173:1181 arg1 termini 1143:1149 the termini 1139:1149 the termini of both N-glycans and O-glycans 1139:1181 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 6 1 theme efficient 932:940 arg1 removal 952:958 efficient enzymatic removal 932:958 efficient enzymatic removal of its N-glycans 932:975 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 1 2 theme non-collagenous 137:151 arg1 sialoprotein 103:114 The highly glycosylated bone sialoprotein 74:114 The highly glycosylated bone sialoprotein (BSP) 74:120 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 2 theme non-collagenous 137:151 arg1 phosphoprotein 153:166 an abundant non-collagenous phosphoprotein 125:166 an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo 125:261 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 6 3 theme MC3T3-E1 870:877 arg1 cells 879:883 MC3T3-E1 cells 870:883 MC3T3-E1 cells 870:883 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 0 4 from status 14:19 arg1 mineralization 58:71 mineralization 58:71 mineralization 58:71 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 9 5 theme osteoblast 1490:1499 arg1 differentiation 1501:1515 osteoblast differentiation 1490:1515 osteoblast differentiation 1490:1515 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 4 6 theme mass 545:548 arg1 MS/MS 564:568 MS/MS 564:568 MS/MS 564:568 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 6 theme mass 545:548 arg1 spectrometry 550:561 tandem mass spectrometry 538:561 tandem mass spectrometry (MS/MS) 538:569 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 1 7 gly glycosylated 85:96 arg1 phosphoprotein 153:166 an abundant non-collagenous phosphoprotein 125:166 an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo 125:261 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 7 gly glycosylated 85:96 arg1 BSP 117:119 BSP 117:119 BSP 117:119 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 7 gly glycosylated 85:96 arg1 sialoprotein 103:114 The highly glycosylated bone sialoprotein 74:114 The highly glycosylated bone sialoprotein (BSP) 74:120 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 6 8 theme cells 879:883 arg1 mineralization 852:865 mineralization 852:865 mineralization 852:865 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 6 8 theme cells 879:883 arg1 differentiation 832:846 osteoblast differentiation 821:846 osteoblast differentiation 821:846 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 1 9 from phosphoprotein 153:166 arg1 bone 171:174 bone 171:174 bone 171:174 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 2 10 theme structural 277:286 arg1 details 288:294 the structural details 273:294 the structural details of its different glycosylation linkages 273:334 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 4 11 theme integrin 629:636 arg1 rhiBSP 660:665 rhiBSP 660:665 rhiBSP 660:665 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 11 theme integrin 629:636 arg1 sialoprotein 646:657 the recombinant human integrin binding sialoprotein 607:657 the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) 591:666 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 0 12 from role 50:53 arg1 mineralization 58:71 mineralization 58:71 mineralization 58:71 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 8 13 theme VDR 1304:1306 arg1 expression 1308:1317 VDR expression 1304:1317 VDR expression 1304:1317 Moreover, BSP-conjugated SAs may affect mineralization via ERK activation of VDR expression. 28958711 3 14 gly sialylated 506:515 arg1 N-glycans 477:485 the N-glycans 473:485 the N-glycans on BSP 473:492 Previous studies suggested that the O-glycans, but not the N-glycans on BSP, are highly sialylated. 28958711 3 14 gly sialylated 506:515 arg1 O-glycans 454:462 the O-glycans 450:462 the O-glycans 450:462 Previous studies suggested that the O-glycans, but not the N-glycans on BSP, are highly sialylated. 28958711 7 15 contain has 1006:1008 arg2 effect 1020:1025 a greater effect 1010:1025 a greater effect 1010:1025 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 7 15 contain has 1006:1008 arg1 Removal 978:984 Removal 978:984 Removal of all terminal SAs 978:1004 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 7 16 theme rhiBSP 1054:1059 arg1 effect 1044:1049 the effect 1040:1049 the effect of rhiBSP on osteogenesis 1040:1075 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 9 17 theme mineralization 1521:1534 arg1 regulation 1476:1485 BSP regulation 1472:1485 BSP regulation of osteoblast differentiation and mineralization 1472:1534 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 2 18 theme bone 386:389 arg1 homeostasis 391:401 bone homeostasis 386:401 bone homeostasis 386:401 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 9 19 from N- 1437:1438 arg1 SAs 1425:1427 SAs 1425:1427 SAs at both N- and O-glycans 1425:1452 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 4 20 theme human 623:627 arg1 rhiBSP 660:665 rhiBSP 660:665 rhiBSP 660:665 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 20 theme human 623:627 arg1 sialoprotein 646:657 the recombinant human integrin binding sialoprotein 607:657 the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) 591:666 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 5 21 theme novel 754:758 arg1 sites 760:764 multiple novel sites 745:764 multiple novel sites of N-glycan modification 745:789 We also identified multiple novel sites of N-glycan modification. 28958711 5 21 theme novel 754:758 arg1 modification 778:789 N-glycan modification 769:789 N-glycan modification 769:789 We also identified multiple novel sites of N-glycan modification. 28958711 7 22 from termini 1143:1149 arg1 sialylation 1124:1134 sialylation 1124:1134 sialylation at the termini of both N-glycans and O-glycans 1124:1181 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 2 23 theme different 303:311 arg1 linkages 327:334 its different glycosylation linkages 299:334 its different glycosylation linkages 299:334 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 0 24 theme Glycosylation 0:12 arg1 status 14:19 Glycosylation status 0:19 Glycosylation status of bone sialoprotein 0:40 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 7 25 from effect 1044:1049 arg1 osteogenesis 1064:1075 osteogenesis 1064:1075 osteogenesis 1064:1075 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 9 26 from O-glycans 1444:1452 arg1 SAs 1425:1427 SAs 1425:1427 SAs at both N- and O-glycans 1425:1452 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 1 27 theme glycosylated 85:96 arg1 phosphoprotein 153:166 an abundant non-collagenous phosphoprotein 125:166 an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo 125:261 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 27 theme glycosylated 85:96 arg1 BSP 117:119 BSP 117:119 BSP 117:119 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 27 theme glycosylated 85:96 arg1 sialoprotein 103:114 The highly glycosylated bone sialoprotein 74:114 The highly glycosylated bone sialoprotein (BSP) 74:120 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 3 28 theme Previous 418:425 arg1 studies 427:433 Previous studies 418:433 Previous studies 418:433 Previous studies suggested that the O-glycans, but not the N-glycans on BSP, are highly sialylated. 28958711 0 29 theme bone 24:27 arg1 sialoprotein 29:40 bone sialoprotein 24:40 bone sialoprotein 24:40 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 6 30 theme enzymatic 942:950 arg1 removal 952:958 efficient enzymatic removal 932:958 efficient enzymatic removal of its N-glycans 932:975 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 3 31 from N-glycans 477:485 arg1 BSP 490:492 BSP 490:492 BSP 490:492 Previous studies suggested that the O-glycans, but not the N-glycans on BSP, are highly sialylated. 28958711 1 32 theme bone 98:101 arg1 phosphoprotein 153:166 an abundant non-collagenous phosphoprotein 125:166 an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo 125:261 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 32 theme bone 98:101 arg1 BSP 117:119 BSP 117:119 BSP 117:119 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 32 theme bone 98:101 arg1 sialoprotein 103:114 The highly glycosylated bone sialoprotein 74:114 The highly glycosylated bone sialoprotein (BSP) 74:120 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 33 theme osteoblast 191:200 arg1 differentiation 202:216 osteoblast differentiation 191:216 osteoblast differentiation 191:216 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 5 34 theme N-glycan 769:776 arg1 modification 778:789 N-glycan modification 769:789 N-glycan modification 769:789 We also identified multiple novel sites of N-glycan modification. 28958711 4 35 theme tandem 538:543 arg1 MS/MS 564:568 MS/MS 564:568 MS/MS 564:568 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 35 theme tandem 538:543 arg1 spectrometry 550:561 tandem mass spectrometry 538:561 tandem mass spectrometry (MS/MS) 538:569 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 9 36 theme BSP 1472:1474 arg1 regulation 1476:1485 BSP regulation 1472:1485 BSP regulation of osteoblast differentiation and mineralization 1472:1534 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 9 37 from SAs 1385:1387 arg1 rhiBSP 1396:1401 the rhiBSP 1392:1401 the rhiBSP 1392:1401 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 6 38 theme osteoblast 821:830 arg1 differentiation 832:846 osteoblast differentiation 821:846 osteoblast differentiation 821:846 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 4 39 theme binding 638:644 arg1 rhiBSP 660:665 rhiBSP 660:665 rhiBSP 660:665 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 39 theme binding 638:644 arg1 sialoprotein 646:657 the recombinant human integrin binding sialoprotein 607:657 the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) 591:666 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 40 theme recombinant 611:621 arg1 rhiBSP 660:665 rhiBSP 660:665 rhiBSP 660:665 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 40 theme recombinant 611:621 arg1 sialoprotein 646:657 the recombinant human integrin binding sialoprotein 607:657 the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) 591:666 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 0 41 theme sialoprotein 29:40 arg1 status 14:19 Glycosylation status 0:19 Glycosylation status of bone sialoprotein 0:40 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 0 41 theme sialoprotein 29:40 arg1 role 50:53 its role 46:53 its role in mineralization 46:71 Glycosylation status of bone sialoprotein and its role in mineralization. 28958711 6 42 theme N-glycans 967:975 arg1 removal 952:958 efficient enzymatic removal 932:958 efficient enzymatic removal of its N-glycans 932:975 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 8 43 theme BSP-conjugated 1237:1250 arg1 SAs 1252:1254 BSP-conjugated SAs 1237:1254 BSP-conjugated SAs 1237:1254 Moreover, BSP-conjugated SAs may affect mineralization via ERK activation of VDR expression. 28958711 2 44 from functions 373:381 arg1 homeostasis 391:401 bone homeostasis 386:401 bone homeostasis 386:401 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 1 45 theme new 222:224 arg1 deposition 231:240 new bone deposition 222:240 new bone deposition 222:240 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 7 46 theme N-glycans 1159:1167 arg1 termini 1143:1149 the termini 1139:1149 the termini of both N-glycans and O-glycans 1139:1181 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 6 47 theme rhiBSP 805:810 arg1 Treatment 792:800 Treatment 792:800 Treatment of rhiBSP 792:810 Treatment of rhiBSP enhances osteoblast differentiation and mineralization of MC3T3-E1 cells and this effect could be partially reversed by efficient enzymatic removal of its N-glycans. 28958711 4 48 dep N-glycanson 595:605 arg1 rhiBSP 660:665 rhiBSP 660:665 rhiBSP 660:665 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 48 dep N-glycanson 595:605 arg1 sialoprotein 646:657 the recombinant human integrin binding sialoprotein 607:657 the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) 591:666 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 7 49 theme terminal 993:1000 arg1 SAs 1002:1004 all terminal SAs 989:1004 all terminal SAs 989:1004 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 5 50 theme multiple 745:752 arg1 sites 760:764 multiple novel sites 745:764 multiple novel sites of N-glycan modification 745:789 We also identified multiple novel sites of N-glycan modification. 28958711 5 50 theme multiple 745:752 arg1 modification 778:789 N-glycan modification 769:789 N-glycan modification 769:789 We also identified multiple novel sites of N-glycan modification. 28958711 7 51 theme greater 1012:1018 arg1 effect 1020:1025 a greater effect 1010:1025 a greater effect 1010:1025 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 9 52 theme differentiation 1501:1515 arg1 regulation 1476:1485 BSP regulation 1472:1485 BSP regulation of osteoblast differentiation and mineralization 1472:1534 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 7 53 theme SAs 1002:1004 arg1 Removal 978:984 Removal 978:984 Removal of all terminal SAs 978:1004 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 2 54 theme linkages 327:334 arg1 details 288:294 the structural details 273:294 the structural details of its different glycosylation linkages 273:334 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 9 55 theme novel 1357:1361 arg1 N-glycans 1363:1371 novel N-glycans 1357:1371 novel N-glycans enriched in SAs on the rhiBSP 1357:1401 Collectively, our results identified novel N-glycans enriched in SAs on the rhiBSP and demonstrated that SAs at both N- and O-glycans are important for BSP regulation of osteoblast differentiation and mineralization in vitro. 28958711 1 56 theme bone 226:229 arg1 deposition 231:240 new bone deposition 222:240 new bone deposition 222:240 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 8 57 theme expression 1308:1317 arg1 activation 1290:1299 ERK activation 1286:1299 ERK activation of VDR expression 1286:1317 Moreover, BSP-conjugated SAs may affect mineralization via ERK activation of VDR expression. 28958711 2 58 theme glycosylation 313:325 arg1 linkages 327:334 its different glycosylation linkages 299:334 its different glycosylation linkages 299:334 However, the structural details of its different glycosylation linkages have not been well studied and their functions in bone homeostasis are not clear. 28958711 4 59 theme sialic 689:694 arg1 SAs 703:705 SAs 703:705 SAs 703:705 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 4 59 theme sialic 689:694 arg1 acids 696:700 sialic acids 689:700 sialic acids (SAs) 689:706 Herein, we employed tandem mass spectrometry (MS/MS) to demonstrate that the N-glycanson the recombinant human integrin binding sialoprotein (rhiBSP) are also enriched in sialic acids (SAs) at their termini. 28958711 5 60 theme modification 778:789 arg1 sites 760:764 multiple novel sites 745:764 multiple novel sites of N-glycan modification 745:789 We also identified multiple novel sites of N-glycan modification. 28958711 5 60 theme modification 778:789 arg1 modification 778:789 N-glycan modification 769:789 N-glycan modification 769:789 We also identified multiple novel sites of N-glycan modification. 28958711 8 61 theme ERK 1286:1288 arg1 activation 1290:1299 ERK activation 1286:1299 ERK activation of VDR expression 1286:1317 Moreover, BSP-conjugated SAs may affect mineralization via ERK activation of VDR expression. 28958711 7 62 theme important 1192:1200 arg1 role 1202:1205 an important role 1189:1205 an important role 1189:1205 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 7 63 gly sialylation 1124:1134 arg1 termini 1143:1149 the termini 1139:1149 the termini of both N-glycans and O-glycans 1139:1181 Removal of all terminal SAs has a greater effect in reversing the effect of rhiBSP on osteogenesis, especially on mineralization, suggesting that sialylation at the termini of both N-glycans and O-glycans plays an important role in this regulation. 28958711 1 64 theme abundant 128:135 arg1 sialoprotein 103:114 The highly glycosylated bone sialoprotein 74:114 The highly glycosylated bone sialoprotein (BSP) 74:120 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 28958711 1 64 theme abundant 128:135 arg1 phosphoprotein 153:166 an abundant non-collagenous phosphoprotein 125:166 an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo 125:261 The highly glycosylated bone sialoprotein (BSP) is an abundant non-collagenous phosphoprotein in bone which enhances osteoblast differentiation and new bone deposition in vitro and in vivo. 29224385 4 0 theme exact 533:537 arg1 process 558:564 the exact pathophysiological process 529:564 the exact pathophysiological process 529:564 However, the exact pathophysiological process is not well understood. 29224385 11 1 theme larger 1769:1774 arg1 cohort 1776:1781 a larger cohort 1767:1781 a larger cohort 1767:1781 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 7 2 theme electrospray 1158:1169 arg1 ionization-Triple-Quadruple 1171:1197 electrospray ionization-Triple-Quadruple 1158:1197 electrospray ionization-Triple-Quadruple 1158:1197 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 9 3 theme fucosylated 1528:1538 arg1 N-glycans 1540:1548 fucosylated N-glycans 1528:1548 fucosylated N-glycans 1528:1548 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 6 4 theme osteonecrosis 893:905 arg1 patients 927:934 steroid-induced osteonecrosis of the femoral head patients 877:934 steroid-induced osteonecrosis of the femoral head patients 877:934 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 7 5 theme flight 1147:1152 arg1 ionization-Q-time 1126:1142 electrospray ionization-Q-time 1113:1142 electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple 1113:1197 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 3 6 theme femoral 506:512 arg1 head 514:517 the femoral head 502:517 the femoral head 502:517 There are over 30 million patients in the US taking steroids, and roughly 40% will develop osteonecrosis of the femoral head. 29224385 1 7 theme femoral 117:123 arg1 head 125:128 the femoral head 113:128 the femoral head 113:128 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 5 8 theme chemical 752:759 arg1 data 761:764 more chemical data 747:764 more chemical data for pathophysiology research and possibly biomarker discovery 747:826 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 8 9 theme whole 1251:1255 arg1 analysis 1277:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 6 10 theme serum 858:862 arg1 samples 864:870 27 serum samples 855:870 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 855:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 5 11 theme serum 635:639 arg1 glycosylation 641:653 serum glycosylation 635:653 serum glycosylation of osteonecrosis of the femoral head 635:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 8 12 theme reaction 1332:1339 arg1 monitoring 1341:1350 multiple reaction monitoring 1323:1350 multiple reaction monitoring 1323:1350 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 10 13 theme head 1741:1744 arg1 osteonecrosis 1712:1724 osteonecrosis 1712:1724 osteonecrosis of the femoral head 1712:1744 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 8 14 theme glycosylation 1263:1275 arg1 analysis 1277:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 9 15 theme osteonecrosis 1589:1601 arg1 development 1574:1584 the development 1570:1584 the development of osteonecrosis of the femoral head 1570:1621 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 1 16 theme head 125:128 arg1 Osteonecrosis 96:108 Osteonecrosis 96:108 Osteonecrosis of the femoral head 96:128 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 1 16 theme head 125:128 arg1 disease 163:169 a recalcitrant and paralyzing disease 133:169 a recalcitrant and paralyzing disease 133:169 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 5 17 theme femoral 679:685 arg1 head 687:690 the femoral head 675:690 the femoral head 675:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 8 18 theme N-glycan 1297:1304 arg1 compositions 1306:1317 14 N-glycan compositions 1294:1317 14 N-glycan compositions 1294:1317 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 11 19 from value 1868:1872 arg1 onset 1885:1889 disease onset 1877:1889 disease onset 1877:1889 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 7 20 theme site-specific 1042:1054 arg1 glycosylation 1056:1068 site-specific glycosylation 1042:1068 site-specific glycosylation of immunoglobulins 1042:1087 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 9 21 theme head 1618:1621 arg1 osteonecrosis 1589:1601 osteonecrosis 1589:1601 osteonecrosis of the femoral head 1589:1621 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 6 22 theme head 922:925 arg1 osteonecrosis 893:905 steroid-induced osteonecrosis 877:905 steroid-induced osteonecrosis of the femoral head patients 877:934 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 6 23 contain containing 844:853 arg1 set 840:842 A training set 829:842 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 829:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 6 23 contain containing 844:853 arg2 samples 864:870 27 serum samples 855:870 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 855:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 11 24 theme disease 1877:1883 arg1 onset 1885:1889 disease onset 1877:1889 disease onset 1877:1889 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 7 25 theme serum 1032:1036 arg1 Glycosylation 1009:1021 Glycosylation 1009:1021 Glycosylation of whole serum 1009:1036 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 7 25 theme serum 1032:1036 arg1 glycosylation 1056:1068 site-specific glycosylation 1042:1068 site-specific glycosylation of immunoglobulins 1042:1087 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 0 26 theme head 69:72 arg1 osteonecrosis 40:52 osteonecrosis 40:52 osteonecrosis of the femoral head 40:72 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 0 27 theme mass 77:80 arg1 spectrometry 82:93 mass spectrometry 77:93 mass spectrometry 77:93 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 2 28 theme steroid 381:387 arg1 use 389:391 long-term steroid use 371:391 long-term steroid use 371:391 Osteonecrosis of the femoral head is typically caused by trauma or long-term steroid use. 29224385 5 29 theme state-of-the-art 702:717 arg1 tools 730:734 the state-of-the-art analytical tools 698:734 the state-of-the-art analytical tools 698:734 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 5 30 theme more 747:750 arg1 data 761:764 more chemical data 747:764 more chemical data for pathophysiology research and possibly biomarker discovery 747:826 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 0 31 theme Serum 0:4 arg1 characterization 20:35 Serum glycosylation characterization 0:35 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 0:94 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 11 32 from stage 1809:1813 arg1 studies 1754:1760 Future studies 1747:1760 Future studies with a larger cohort 1747:1781 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 11 32 from stage 1809:1813 arg1 patients 1787:1794 patients 1787:1794 patients from earlier stage 1787:1813 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 6 33 from gender- 948:954 arg1 controls 972:979 age-matched controls 960:979 age-matched controls 960:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 6 33 from gender- 948:954 arg1 patients 927:934 steroid-induced osteonecrosis of the femoral head patients 877:934 steroid-induced osteonecrosis of the femoral head patients 877:934 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 7 34 theme reaction 1212:1219 arg1 monitoring 1221:1230 multiple reaction monitoring 1203:1230 multiple reaction monitoring 1203:1230 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 1 35 theme diagnostic 284:293 arg1 imaging 295:301 diagnostic imaging 284:301 diagnostic imaging 284:301 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 0 36 theme osteonecrosis 40:52 arg1 characterization 20:35 Serum glycosylation characterization 0:35 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 0:94 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 7 37 gly Glycosylation 1009:1021 arg1 immunoglobulins 1073:1087 immunoglobulins 1073:1087 immunoglobulins 1073:1087 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 7 37 gly Glycosylation 1009:1021 arg1 serum 1032:1036 whole serum 1026:1036 whole serum 1026:1036 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 10 38 theme protein 1661:1667 arg1 modification 1669:1680 a posttranslational protein modification 1641:1680 a posttranslational protein modification 1641:1680 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 10 38 theme protein 1661:1667 arg1 Glycosylation 1624:1636 Glycosylation 1624:1636 Glycosylation 1624:1636 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 2 39 theme head 333:336 arg1 Osteonecrosis 304:316 Osteonecrosis 304:316 Osteonecrosis of the femoral head 304:336 Osteonecrosis of the femoral head is typically caused by trauma or long-term steroid use. 29224385 9 40 theme nonsialylated 1472:1484 arg1 N-glycans 1502:1510 nonsialylated, nonfucosylated N-glycans 1472:1510 nonsialylated, nonfucosylated N-glycans 1472:1510 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 6 41 from 25 940:941 arg1 samples 864:870 27 serum samples 855:870 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 855:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 8 42 with controls 1416:1423 arg1 significance 1442:1453 statistical significance 1430:1453 statistical significance 1430:1453 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 9 43 gly fucosylated 1528:1538 arg1 N-glycans 1540:1548 fucosylated N-glycans 1528:1548 fucosylated N-glycans 1528:1548 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 11 44 with studies 1754:1760 arg1 cohort 1776:1781 a larger cohort 1767:1781 a larger cohort 1767:1781 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 8 45 with cases 1406:1410 arg1 significance 1442:1453 statistical significance 1430:1453 statistical significance 1430:1453 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 1 46 theme diagnosis 220:228 arg1 time 212:215 the time 208:215 the time of diagnosis 208:228 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 0 47 theme femoral 61:67 arg1 head 69:72 the femoral head 57:72 the femoral head 57:72 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 6 48 theme femoral 914:920 arg1 head 922:925 the femoral head 910:925 the femoral head 910:925 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 6 49 from patients 927:934 arg1 samples 864:870 27 serum samples 855:870 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 855:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 11 50 with patients 1787:1794 arg1 cohort 1776:1781 a larger cohort 1767:1781 a larger cohort 1767:1781 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 9 51 theme N-glycans 1540:1548 arg1 decrease 1516:1523 decrease 1516:1523 decrease of fucosylated N-glycans 1516:1548 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 9 51 theme N-glycans 1540:1548 arg1 increase 1460:1467 The increase 1456:1467 The increase of nonsialylated, nonfucosylated N-glycans 1456:1510 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 6 52 theme steroid-induced 877:891 arg1 osteonecrosis 893:905 steroid-induced osteonecrosis 877:905 steroid-induced osteonecrosis of the femoral head patients 877:934 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 4 53 theme pathophysiological 539:556 arg1 process 558:564 the exact pathophysiological process 529:564 the exact pathophysiological process 529:564 However, the exact pathophysiological process is not well understood. 29224385 7 54 theme electrospray 1113:1124 arg1 ionization-Q-time 1126:1142 electrospray ionization-Q-time 1113:1142 electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple 1113:1197 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 11 55 theme earlier 1801:1807 arg1 stage 1809:1813 earlier stage 1801:1813 earlier stage 1801:1813 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 6 56 theme age-matched 960:970 arg1 controls 972:979 age-matched controls 960:979 age-matched controls 960:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 8 57 theme multiple 1323:1330 arg1 monitoring 1341:1350 multiple reaction monitoring 1323:1350 multiple reaction monitoring 1323:1350 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 10 58 theme femoral 1733:1739 arg1 head 1741:1744 the femoral head 1729:1744 the femoral head 1729:1744 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 7 59 theme immunoglobulins 1073:1087 arg1 Glycosylation 1009:1021 Glycosylation 1009:1021 Glycosylation of whole serum 1009:1036 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 7 59 theme immunoglobulins 1073:1087 arg1 glycosylation 1056:1068 site-specific glycosylation 1042:1068 site-specific glycosylation of immunoglobulins 1042:1087 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 3 60 theme head 514:517 arg1 osteonecrosis 485:497 osteonecrosis 485:497 osteonecrosis of the femoral head 485:517 There are over 30 million patients in the US taking steroids, and roughly 40% will develop osteonecrosis of the femoral head. 29224385 1 61 theme end 195:197 arg1 stage 199:203 the end stage 191:203 the end stage 191:203 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 8 62 theme serum 1257:1261 arg1 analysis 1277:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis 1247:1284 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 7 63 gly glycosylation 1056:1068 arg1 immunoglobulins 1073:1087 immunoglobulins 1073:1087 immunoglobulins 1073:1087 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 7 63 gly glycosylation 1056:1068 arg1 serum 1032:1036 whole serum 1026:1036 whole serum 1026:1036 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 9 64 theme femoral 1610:1616 arg1 head 1618:1621 the femoral head 1606:1621 the femoral head 1606:1621 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 5 65 theme osteonecrosis 658:670 arg1 glycosylation 641:653 serum glycosylation 635:653 serum glycosylation of osteonecrosis of the femoral head 635:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 7 66 theme whole 1026:1030 arg1 serum 1032:1036 whole serum 1026:1036 whole serum 1026:1036 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 5 67 theme analytical 719:728 arg1 tools 730:734 the state-of-the-art analytical tools 698:734 the state-of-the-art analytical tools 698:734 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 8 68 gly glycopeptides 1366:1378 arg2 glycopeptides 1366:1378 eight glycopeptides 1360:1378 eight glycopeptides that were altered between cases and controls with statistical significance 1360:1453 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 5 69 theme biomarker 808:816 arg1 discovery 818:826 biomarker discovery 808:826 biomarker discovery 808:826 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 1 70 theme recalcitrant 135:146 arg1 Osteonecrosis 96:108 Osteonecrosis 96:108 Osteonecrosis of the femoral head 96:128 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 1 70 theme recalcitrant 135:146 arg1 disease 163:169 a recalcitrant and paralyzing disease 133:169 a recalcitrant and paralyzing disease 133:169 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 1 71 theme paralyzing 152:161 arg1 Osteonecrosis 96:108 Osteonecrosis 96:108 Osteonecrosis of the femoral head 96:128 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 1 71 theme paralyzing 152:161 arg1 disease 163:169 a recalcitrant and paralyzing disease 133:169 a recalcitrant and paralyzing disease 133:169 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 5 72 gly glycosylation 641:653 arg1 osteonecrosis 658:670 osteonecrosis 658:670 osteonecrosis of the femoral head 658:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 5 73 theme head 687:690 arg1 osteonecrosis 658:670 osteonecrosis 658:670 osteonecrosis of the femoral head 658:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 9 74 dep nonsialylated 1472:1484 arg1 nonfucosylated 1487:1500 nonfucosylated 1487:1500 nonfucosylated 1487:1500 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 8 75 theme statistical 1430:1440 arg1 significance 1442:1453 statistical significance 1430:1453 statistical significance 1430:1453 The whole serum glycosylation analysis yielded 14 N-glycan compositions and multiple reaction monitoring yielded eight glycopeptides that were altered between cases and controls with statistical significance. 29224385 6 76 from controls 972:979 arg1 samples 864:870 27 serum samples 855:870 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 855:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 2 77 theme long-term 371:379 arg1 use 389:391 long-term steroid use 371:391 long-term steroid use 371:391 Osteonecrosis of the femoral head is typically caused by trauma or long-term steroid use. 29224385 0 78 theme glycosylation 6:18 arg1 characterization 20:35 Serum glycosylation characterization 0:35 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 0:94 Serum glycosylation characterization of osteonecrosis of the femoral head by mass spectrometry. 29224385 11 79 theme potential 1849:1857 arg1 markers 1859:1865 these potential markers 1843:1865 these potential markers' value in disease onset 1843:1889 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 1 80 theme physical 259:266 arg1 examination 268:278 physical examination 259:278 physical examination 259:278 Osteonecrosis of the femoral head is a recalcitrant and paralyzing disease often discovered in the end stage at the time of diagnosis, which is often performed by physical examination and diagnostic imaging. 29224385 9 81 gly nonsialylated 1472:1484 arg1 N-glycans 1502:1510 nonsialylated, nonfucosylated N-glycans 1472:1510 nonsialylated, nonfucosylated N-glycans 1472:1510 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 7 82 theme multiple 1203:1210 arg1 monitoring 1221:1230 multiple reaction monitoring 1203:1230 multiple reaction monitoring 1203:1230 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 3 83 theme million 412:418 arg1 patients 420:427 30 million patients 409:427 30 million patients 409:427 There are over 30 million patients in the US taking steroids, and roughly 40% will develop osteonecrosis of the femoral head. 29224385 7 84 theme ionization-Triple-Quadruple 1171:1197 arg1 ionization-Q-time 1126:1142 electrospray ionization-Q-time 1113:1142 electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple 1113:1197 Glycosylation of whole serum and site-specific glycosylation of immunoglobulins are characterized using electrospray ionization-Q-time of flight and electrospray ionization-Triple-Quadruple via multiple reaction monitoring, respectively. 29224385 5 85 from alteration 621:630 arg1 glycosylation 641:653 serum glycosylation 635:653 serum glycosylation of osteonecrosis of the femoral head 635:690 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 29224385 6 86 theme training 831:838 arg1 set 840:842 A training set 829:842 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls 829:979 A training set containing 27 serum samples from steroid-induced osteonecrosis of the femoral head patients and 25 from gender- and age-matched controls was collected and analyzed. 29224385 10 87 theme posttranslational 1643:1659 arg1 modification 1669:1680 a posttranslational protein modification 1641:1680 a posttranslational protein modification 1641:1680 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 10 87 theme posttranslational 1643:1659 arg1 Glycosylation 1624:1636 Glycosylation 1624:1636 Glycosylation 1624:1636 Glycosylation is a posttranslational protein modification and is apparently affected by osteonecrosis of the femoral head. 29224385 2 88 theme femoral 325:331 arg1 head 333:336 the femoral head 321:336 the femoral head 321:336 Osteonecrosis of the femoral head is typically caused by trauma or long-term steroid use. 29224385 9 89 theme N-glycans 1502:1510 arg1 decrease 1516:1523 decrease 1516:1523 decrease of fucosylated N-glycans 1516:1548 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 9 89 theme N-glycans 1502:1510 arg1 increase 1460:1467 The increase 1456:1467 The increase of nonsialylated, nonfucosylated N-glycans 1456:1510 The increase of nonsialylated, nonfucosylated N-glycans and decrease of fucosylated N-glycans are associated with the development of osteonecrosis of the femoral head. 29224385 11 90 theme Future 1747:1752 arg1 studies 1754:1760 Future studies 1747:1760 Future studies with a larger cohort 1747:1781 Future studies with a larger cohort and patients from earlier stage will be performed to assess these potential markers' value in disease onset. 29224385 5 91 theme pathophysiology 770:784 arg1 research 786:793 pathophysiology research 770:793 pathophysiology research 770:793 This study aims to examine the alteration in serum glycosylation of osteonecrosis of the femoral head using the state-of-the-art analytical tools to provide more chemical data for pathophysiology research and possibly biomarker discovery. 27356208 10 0 theme suggested 1647:1655 arg1 analogs 1657:1663 five suggested analogs 1642:1663 five suggested analogs 1642:1663 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 9 1 theme relative 1593:1600 arg1 substitutions 1578:1590 amino acid substitutions 1567:1590 amino acid substitutions 1567:1590 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 3 2 theme proteins 570:577 arg1 hyper-glycosylation 535:553 the hyper-glycosylation 531:553 the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods 531:633 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 6 3 theme new 1105:1107 arg1 s 1130:1130 s 1130:1130 s 1130:1130 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 6 3 theme new 1105:1107 arg1 site 1125:1128 new N-glycosylation site 1105:1128 new N-glycosylation site(s) 1105:1131 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 9 4 theme structures 1368:1377 arg1 generation 1336:1345 generation 1336:1345 generation of three-dimensional structures 1336:1377 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 11 5 theme experimental 2025:2036 arg1 investigations 2038:2051 further experimental investigations 2017:2051 further experimental investigations 2017:2051 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 4 6 from selection 652:660 arg1 surface 692:698 the surface 688:698 the surface of a protein for introduction of a new N-glycosylation site 688:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 1 7 theme asparagine 161:170 arg1 residue 172:178 the asparagine residue 157:178 the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline 157:286 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 3 8 theme recombinant 558:568 arg1 proteins 570:577 recombinant proteins 558:577 recombinant proteins 558:577 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 5 9 theme suitable 842:849 arg1 s 860:860 suitable location(s) 842:861 suitable location(s) 842:861 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 11 10 theme similar 1922:1928 arg1 structure 1930:1938 the most similar structure 1913:1938 the most similar structure 1913:1938 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 5 11 theme computational 795:807 arg1 approach 809:816 a computational approach 793:816 a computational approach 793:816 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 11 12 theme dynamic 1944:1950 arg1 behavior 1952:1959 dynamic behavior 1944:1959 dynamic behavior 1944:1959 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 9 13 theme designed 1417:1424 arg1 analogs 1426:1432 the five designed analogs 1408:1432 the five designed analogs 1408:1432 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 11 14 theme native 1976:1981 arg1 counterpart 1983:1993 the native counterpart 1972:1993 the native counterpart 1972:1993 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 3 15 theme mutagenesis 615:625 arg1 methods 627:633 the site-directed mutagenesis methods 597:633 the site-directed mutagenesis methods 597:633 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 4 16 theme protein 705:711 arg1 surface 692:698 the surface 688:698 the surface of a protein for introduction of a new N-glycosylation site 688:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 3 17 theme methods 627:633 arg1 application 582:592 application 582:592 application of the site-directed mutagenesis methods 582:633 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 1 18 from residue 172:178 arg1 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 1 18 from residue 172:178 arg1 sequence 213:220 the N-glycosylation consensus sequence 183:220 the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr) 183:238 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 5 19 theme N-glycosylation 883:897 arg1 sites 899:903 new N-glycosylation sites 879:903 new N-glycosylation sites 879:903 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 3 20 gly hyper-glycosylation 535:553 arg1 proteins 570:577 recombinant proteins 558:577 recombinant proteins 558:577 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 4 21 theme location 676:683 arg1 concern 770:776 a main concern 763:776 a main concern 763:776 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 4 21 theme location 676:683 arg1 selection 652:660 selection 652:660 selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site 652:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 4 22 gly N-glycosylation 739:753 arg2 site 755:758 a new N-glycosylation site 733:758 a new N-glycosylation site 733:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 3 23 theme site-directed 601:613 arg1 mutagenesis 615:625 site-directed mutagenesis 601:625 the site-directed mutagenesis methods 597:633 Glyco-engineering is an approach to facilitate the hyper-glycosylation of recombinant proteins by application of the site-directed mutagenesis methods. 27356208 9 24 theme dynamic 1461:1467 arg1 simulations 1474:1484 molecular dynamic (MD) simulations 1451:1484 molecular dynamic (MD) simulations 1451:1484 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 0 25 theme factor 76:81 arg1 IX 83:84 the human coagulation factor IX 54:84 the human coagulation factor IX 54:84 In silico designing of hyper-glycosylated analogs for the human coagulation factor IX. 27356208 9 26 theme MD 1470:1471 arg1 simulations 1474:1484 molecular dynamic (MD) simulations 1451:1484 molecular dynamic (MD) simulations 1451:1484 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 10 27 theme Root 1749:1752 arg1 Deviation 1766:1774 Root Mean Square Deviation 1749:1774 relatively constant Root Mean Square Deviation values 1729:1781 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 11 28 from analysis 1795:1802 arg1 data 1811:1814 the data 1807:1814 the data obtained during MD simulations 1807:1845 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 4 29 theme N-glycosylation 739:753 arg1 site 755:758 a new N-glycosylation site 733:758 a new N-glycosylation site 733:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 5 30 theme factor 932:937 arg1 hFIX 943:946 hFIX 943:946 hFIX 943:946 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 5 30 theme factor 932:937 arg1 IX 939:940 the human coagulation factor IX 910:940 the human coagulation factor IX (hFIX) 910:947 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 6 31 theme mature 990:995 arg1 hFIX 997:1000 mature hFIX 990:1000 mature hFIX 990:1000 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 2 32 theme protein 410:416 arg1 properties 418:427 the protein properties 406:427 the protein properties such as solubility, folding, stability, and secretion 406:481 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 2 32 theme protein 410:416 arg1 solubility 437:446 solubility 437:446 solubility 437:446 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 2 32 theme protein 410:416 arg1 stability 458:466 stability 458:466 stability 458:466 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 2 32 theme protein 410:416 arg1 secretion 473:481 secretion 473:481 secretion 473:481 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 2 32 theme protein 410:416 arg1 folding 449:455 folding 449:455 folding 449:455 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 8 33 theme N-glycosylation 1291:1305 arg1 prediction 1318:1327 N-glycosylation efficiency prediction 1291:1327 N-glycosylation efficiency prediction 1291:1327 For each suggested position, an analog was defined and subjected for N-glycosylation efficiency prediction. 27356208 5 34 theme human 914:918 arg1 hFIX 943:946 hFIX 943:946 hFIX 943:946 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 5 34 theme human 914:918 arg1 IX 939:940 the human coagulation factor IX 910:940 the human coagulation factor IX (hFIX) 910:947 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 8 35 theme suggested 1231:1239 arg1 position 1241:1248 each suggested position 1226:1248 each suggested position 1226:1248 For each suggested position, an analog was defined and subjected for N-glycosylation efficiency prediction. 27356208 0 36 theme human 58:62 arg1 IX 83:84 the human coagulation factor IX 54:84 the human coagulation factor IX 54:84 In silico designing of hyper-glycosylated analogs for the human coagulation factor IX. 27356208 0 37 theme hyper-glycosylated 23:40 arg1 analogs 42:48 hyper-glycosylated analogs 23:48 hyper-glycosylated analogs for the human coagulation factor IX 23:84 In silico designing of hyper-glycosylated analogs for the human coagulation factor IX. 27356208 11 38 with analog 1901:1906 arg1 structure 1930:1938 the most similar structure 1913:1938 the most similar structure 1913:1938 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 11 38 with analog 1901:1906 arg1 behavior 1952:1959 dynamic behavior 1944:1959 dynamic behavior 1944:1959 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 9 39 theme probable 1525:1532 arg1 distortions 1545:1555 probable structural distortions 1525:1555 probable structural distortions caused by amino acid substitutions, relative to the native counterpart 1525:1626 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 6 40 theme first 969:973 arg1 residues 978:985 the first 45 residues 965:985 the first 45 residues of mature hFIX 965:1000 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 6 40 theme first 969:973 arg1 hFIX 997:1000 mature hFIX 990:1000 mature hFIX 990:1000 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 5 41 theme location 851:858 arg1 s 860:860 suitable location(s) 842:861 suitable location(s) 842:861 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 2 42 gly N-glycosylation 311:325 arg2 site 327:330 a new N-glycosylation site 305:330 a new N-glycosylation site 305:330 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 10 43 theme Square 1759:1764 arg1 Deviation 1766:1774 Root Mean Square Deviation 1749:1774 relatively constant Root Mean Square Deviation values 1729:1781 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 2 44 theme site 327:330 arg1 Introduction 289:300 Introduction 289:300 Introduction of a new N-glycosylation site into a protein backbone 289:354 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 4 45 theme main 765:768 arg1 concern 770:776 a main concern 763:776 a main concern 763:776 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 4 45 theme main 765:768 arg1 selection 652:660 selection 652:660 selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site 652:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 2 46 theme new 307:309 arg1 site 327:330 a new N-glycosylation site 305:330 a new N-glycosylation site 305:330 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 7 47 theme potential 1176:1184 arg1 positions 1186:1194 five potential positions 1171:1194 five potential positions 1171:1194 Our exploration lead to detection of five potential positions, for hyper-glycosylation. 27356208 9 48 theme acid 1573:1576 arg1 substitutions 1578:1590 amino acid substitutions 1567:1590 amino acid substitutions 1567:1590 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 6 49 gly N-glycosylation 1109:1123 arg2 s 1130:1130 s 1130:1130 s 1130:1130 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 6 49 gly N-glycosylation 1109:1123 arg2 site 1125:1128 new N-glycosylation site 1105:1128 new N-glycosylation site(s) 1105:1131 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 11 50 theme Additional 1784:1793 arg1 analysis 1795:1802 Additional analysis 1784:1802 Additional analysis 1784:1802 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 9 51 theme three-dimensional 1350:1366 arg1 structures 1368:1377 three-dimensional structures 1350:1377 three-dimensional structures 1350:1377 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 11 52 theme qualified 1891:1899 arg1 analog 1901:1906 the only qualified analog 1882:1906 the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart 1882:1993 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 11 52 theme qualified 1891:1899 arg1 R37N 1874:1877 R37N 1874:1877 R37N 1874:1877 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 9 53 theme native 1609:1614 arg1 counterpart 1616:1626 the native counterpart 1605:1626 the native counterpart 1605:1626 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 1 54 attach attaches 145:152 arg1 residue 172:178 the asparagine residue 157:178 the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline 157:286 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 1 54 attach attaches 145:152 arg2 moiety 138:143 a glycan moiety 129:143 a glycan moiety 129:143 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 9 55 theme homology-based 1383:1396 arg1 modeling 1398:1405 homology-based modeling 1383:1405 homology-based modeling 1383:1405 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 11 56 theme further 2017:2023 arg1 investigations 2038:2051 further experimental investigations 2017:2051 further experimental investigations 2017:2051 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 6 57 theme Ser/Thr 1077:1083 arg1 residues 1085:1092 either Asn or Ser/Thr residues 1063:1092 residues 1085:1092 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 1 58 theme glycan 131:136 arg1 moiety 138:143 a glycan moiety 129:143 a glycan moiety 129:143 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 1 59 theme consensus 203:211 arg1 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 1 59 theme consensus 203:211 arg1 sequence 213:220 the N-glycosylation consensus sequence 183:220 the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr) 183:238 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 10 60 theme equilibration 1704:1716 arg1 state 1718:1722 equilibration state 1704:1722 equilibration state 1704:1722 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 10 61 theme Mean 1754:1757 arg1 Deviation 1766:1774 Root Mean Square Deviation 1749:1774 relatively constant Root Mean Square Deviation values 1729:1781 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 5 62 theme new 879:881 arg1 sites 899:903 new N-glycosylation sites 879:903 new N-glycosylation sites 879:903 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 0 63 theme coagulation 64:74 arg1 IX 83:84 the human coagulation factor IX 54:84 the human coagulation factor IX 54:84 In silico designing of hyper-glycosylated analogs for the human coagulation factor IX. 27356208 10 64 theme constant 1740:1747 arg1 values 1776:1781 relatively constant Root Mean Square Deviation values 1729:1781 relatively constant Root Mean Square Deviation values 1729:1781 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 4 65 from concern 770:776 arg1 regard 644:649 this regard 639:649 this regard 639:649 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 9 66 theme molecular 1451:1459 arg1 simulations 1474:1484 molecular dynamic (MD) simulations 1451:1484 molecular dynamic (MD) simulations 1451:1484 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 6 67 theme N-glycosylation 1109:1123 arg1 s 1130:1130 s 1130:1130 s 1130:1130 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 6 67 theme N-glycosylation 1109:1123 arg1 site 1125:1128 new N-glycosylation site 1105:1128 new N-glycosylation site(s) 1105:1131 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 4 68 theme suitable 667:674 arg1 location 676:683 a suitable location 665:683 a suitable location 665:683 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 5 69 theme coagulation 920:930 arg1 hFIX 943:946 hFIX 943:946 hFIX 943:946 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 5 69 theme coagulation 920:930 arg1 IX 939:940 the human coagulation factor IX 910:940 the human coagulation factor IX (hFIX) 910:947 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 4 70 theme new 735:737 arg1 site 755:758 a new N-glycosylation site 733:758 a new N-glycosylation site 733:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 1 71 theme N-glycosylation 187:201 arg1 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 Asn-Xxx-Ser/Thr 223:237 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 1 71 theme N-glycosylation 187:201 arg1 sequence 213:220 the N-glycosylation consensus sequence 183:220 the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr) 183:238 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 4 72 theme site 755:758 arg1 introduction 717:728 introduction 717:728 introduction of a new N-glycosylation site 717:758 In this regard, selection of a suitable location on the surface of a protein for introduction of a new N-glycosylation site is a main concern. 27356208 11 73 theme MD 1832:1833 arg1 simulations 1835:1845 MD simulations 1832:1845 MD simulations 1832:1845 Additional analysis on the data obtained during MD simulations, lead us to conclude that, R37N is the only qualified analog with the most similar structure and dynamic behavior to that of the native counterpart, to be considered for further experimental investigations. 27356208 6 74 theme hFIX 997:1000 arg1 residues 978:985 the first 45 residues 965:985 the first 45 residues of mature hFIX 965:1000 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 6 74 theme hFIX 997:1000 arg1 hFIX 997:1000 mature hFIX 990:1000 mature hFIX 990:1000 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 5 75 gly N-glycosylation 883:897 arg2 sites 899:903 new N-glycosylation sites 879:903 new N-glycosylation sites 879:903 In this work, a computational approach was conducted to select suitable location(s) for introducing new N-glycosylation sites into the human coagulation factor IX (hFIX). 27356208 10 76 dep Three 1629:1633 arg1 K22N 1680:1683 K22N 1680:1683 K22N 1680:1683 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 10 76 dep Three 1629:1633 arg1 R37N 1690:1693 R37N 1690:1693 R37N 1690:1693 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 10 76 dep Three 1629:1633 arg1 E15T 1674:1677 E15T 1674:1677 E15T 1674:1677 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 1 77 theme amino 262:266 arg1 acid 268:271 any amino acid 258:271 any amino acid 258:271 N-glycosylation is a process during which a glycan moiety attaches to the asparagine residue in the N-glycosylation consensus sequence (Asn-Xxx-Ser/Thr), where Xxx can be any amino acid except proline. 27356208 9 78 theme stability 1504:1512 arg1 levels 1514:1519 their stability levels 1498:1519 their stability levels 1498:1519 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 10 79 theme Deviation 1766:1774 arg1 values 1776:1781 relatively constant Root Mean Square Deviation values 1729:1781 relatively constant Root Mean Square Deviation values 1729:1781 Three out of five suggested analogs, namely; E15T, K22N, and R37N, reached equilibration state with relatively constant Root Mean Square Deviation values. 27356208 8 80 theme efficiency 1307:1316 arg1 prediction 1318:1327 N-glycosylation efficiency prediction 1291:1327 N-glycosylation efficiency prediction 1291:1327 For each suggested position, an analog was defined and subjected for N-glycosylation efficiency prediction. 27356208 9 81 theme structural 1534:1543 arg1 distortions 1545:1555 probable structural distortions 1525:1555 probable structural distortions caused by amino acid substitutions, relative to the native counterpart 1525:1626 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 7 82 theme positions 1186:1194 arg1 detection 1158:1166 detection 1158:1166 detection of five potential positions 1158:1194 Our exploration lead to detection of five potential positions, for hyper-glycosylation. 27356208 2 83 theme N-glycosylation 311:325 arg1 site 327:330 a new N-glycosylation site 305:330 a new N-glycosylation site 305:330 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 27356208 0 84 gly hyper-glycosylated 23:40 arg1 analogs 42:48 hyper-glycosylated analogs 23:48 hyper-glycosylated analogs for the human coagulation factor IX 23:84 In silico designing of hyper-glycosylated analogs for the human coagulation factor IX. 27356208 9 85 theme amino 1567:1571 arg1 substitutions 1578:1590 amino acid substitutions 1567:1590 amino acid substitutions 1567:1590 After generation of three-dimensional structures, by homology-based modeling, the five designed analogs were examined by molecular dynamic (MD) simulations, to predict their stability levels and probable structural distortions caused by amino acid substitutions, relative to the native counterpart. 27356208 6 86 theme suitable 1028:1035 arg1 positions 1037:1045 suitable positions 1028:1045 suitable positions 1028:1045 With this aim, the first 45 residues of mature hFIX were explored to find out suitable positions for introducing either Asn or Ser/Thr residues, to create new N-glycosylation site(s). 27356208 2 87 theme protein 339:345 arg1 backbone 347:354 a protein backbone 337:354 a protein backbone 337:354 Introduction of a new N-glycosylation site into a protein backbone leads to its hyper-glycosylation, and may improve the protein properties such as solubility, folding, stability, and secretion. 26797772 8 0 theme N-glycans 1475:1483 arg1 devoid 1456:1461 devoid 1456:1461 devoid 1456:1461 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 9 1 from TNSALP 1681:1686 arg1 series 1632:1637 a series 1630:1637 a series of multiple N-glycan depletion mutants in TNSALP 1630:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 6 2 theme N430D 1105:1109 arg1 mutant 1112:1117 a TNSALP (N430D) mutant 1095:1117 a TNSALP (N430D) mutant 1095:1117 However, we found that TNSALP (N430Q) and TNSALP (N430E) mutants, but not a TNSALP (N430D) mutant, failed to form dimers. 26797772 8 3 theme transfected 1546:1556 arg1 cells 1558:1562 transfected cells 1546:1562 transfected cells 1546:1562 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 9 4 from mutants 1670:1676 arg1 TNSALP 1681:1686 TNSALP 1681:1686 TNSALP 1681:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 6 5 theme TNSALP 1097:1102 arg1 mutant 1112:1117 a TNSALP (N430D) mutant 1095:1117 a TNSALP (N430D) mutant 1095:1117 However, we found that TNSALP (N430Q) and TNSALP (N430E) mutants, but not a TNSALP (N430D) mutant, failed to form dimers. 26797772 4 6 with patient 656:662 arg1 HPP 679:681 infantile HPP 669:681 infantile HPP 669:681 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 9 7 theme N-glycan 1651:1658 arg1 mutants 1670:1676 multiple N-glycan depletion mutants 1642:1676 multiple N-glycan depletion mutants in TNSALP 1642:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 5 8 theme TNSALP 847:852 arg1 mutants 836:842 individual single N-glycan deletion mutants 800:842 individual single N-glycan deletion mutants of TNSALP 800:852 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 7 9 theme N430E 1283:1287 arg1 mutants 1290:1296 TNSALP (N430E) mutants 1275:1296 TNSALP (N430E) mutants 1275:1296 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 5 10 dep structure 987:995 arg1 the 983:985 the 983:985 the 983:985 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 3 11 theme standard 566:573 arg1 nomenclature 575:586 standard nomenclature 566:586 standard nomenclature 566:586 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 8 12 located present 1489:1495 arg1 cells 1558:1562 transfected cells 1546:1562 transfected cells 1546:1562 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 8 12 located present 1489:1495 arg2 TNSALP 1449:1454 TNSALP 1449:1454 TNSALP devoid of all five N-glycans 1449:1483 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 7 13 theme TNSALP 1275:1280 arg1 mutants 1290:1296 TNSALP (N430E) mutants 1275:1296 TNSALP (N430E) mutants 1275:1296 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 9 14 from expression 1845:1854 arg1 cell 1861:1864 a cell 1859:1864 a cell 1859:1864 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 5 15 theme N-glycan 818:825 arg1 mutants 836:842 individual single N-glycan deletion mutants 800:842 individual single N-glycan deletion mutants of TNSALP 800:852 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 5 16 theme site-directed 690:702 arg1 mutagenesis 704:714 site-directed mutagenesis 690:714 site-directed mutagenesis 690:714 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 5 17 theme TNSALP 1013:1018 arg1 function 1001:1008 function 1001:1008 function 1001:1008 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 5 17 theme TNSALP 1013:1018 arg1 structure 987:995 structure 987:995 structure 987:995 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 9 18 dep structure 1778:1786 arg1 the 1774:1776 the 1774:1776 the 1774:1776 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 1 19 theme alkaline 176:183 arg1 glycoprotein 220:231 a membrane glycoprotein 209:231 a membrane glycoprotein with a proposed role in bone mineralization 209:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 19 theme alkaline 176:183 arg1 TNSALP 198:203 TNSALP 198:203 TNSALP 198:203 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 19 theme alkaline 176:183 arg1 phosphatase 185:195 Tissue-nonspecific alkaline phosphatase 157:195 Tissue-nonspecific alkaline phosphatase (TNSALP) 157:204 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 9 20 theme prerequisite 1817:1828 arg1 function 1792:1799 function 1792:1799 function 1792:1799 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 20 theme prerequisite 1817:1828 arg1 structure 1778:1786 structure 1778:1786 structure 1778:1786 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 21 theme comprehensive 1604:1616 arg1 analysis 1618:1625 A comprehensive analysis 1602:1625 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP 1602:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 2 22 from deficiency 446:455 arg1 activity 470:477 serum ALP activity 460:477 serum ALP activity 460:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 4 23 theme infantile 669:677 arg1 HPP 679:681 infantile HPP 669:681 infantile HPP 669:681 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 9 24 theme series 1632:1637 arg1 analysis 1618:1625 A comprehensive analysis 1602:1625 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP 1602:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 2 25 theme bone 425:428 arg1 deficiency 446:455 a deficiency 444:455 a deficiency in serum ALP activity 444:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 25 theme bone 425:428 arg1 hypomineralization 403:420 hypomineralization 403:420 hypomineralization of bone and teeth 403:438 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 26 theme ALP 466:468 arg1 activity 470:477 serum ALP activity 460:477 serum ALP activity 460:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 0 27 theme Glycosylation-deficient 0:22 arg1 mutations 24:32 Glycosylation-deficient mutations 0:32 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase 0:75 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 9 28 theme depletion 1660:1668 arg1 mutants 1670:1676 multiple N-glycan depletion mutants 1642:1676 multiple N-glycan depletion mutants in TNSALP 1642:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 5 29 theme individual 800:809 arg1 mutants 836:842 individual single N-glycan deletion mutants 800:842 individual single N-glycan deletion mutants of TNSALP 800:852 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 9 30 from series 1632:1637 arg1 TNSALP 1681:1686 TNSALP 1681:1686 TNSALP 1681:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 8 31 theme individual 1404:1413 arg1 mutants 1440:1446 individual single N-glycan deletion mutants 1404:1446 individual single N-glycan deletion mutants 1404:1446 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 2 32 theme teeth 434:438 arg1 deficiency 446:455 a deficiency 444:455 a deficiency in serum ALP activity 444:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 32 theme teeth 434:438 arg1 hypomineralization 403:420 hypomineralization 403:420 hypomineralization of bone and teeth 403:438 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 0 33 theme alkaline 56:63 arg1 phosphatase 65:75 tissue-nonspecific alkaline phosphatase 37:75 tissue-nonspecific alkaline phosphatase 37:75 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 7 34 theme ALP 1375:1377 arg1 activity 1379:1386 strong ALP activity 1368:1386 strong ALP activity 1368:1386 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 2 35 from hypomineralization 403:420 arg1 activity 470:477 serum ALP activity 460:477 serum ALP activity 460:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 8 36 theme N-glycan 1422:1429 arg1 mutants 1440:1446 individual single N-glycan deletion mutants 1404:1446 individual single N-glycan deletion mutants 1404:1446 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 3 37 theme N-glycosylation 506:520 arg1 sites 522:526 five potential N-glycosylation sites 491:526 five potential N-glycosylation sites 491:526 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 5 38 theme COS-1 779:783 arg1 cells 785:789 transfected COS-1 cells 767:789 transfected COS-1 cells 767:789 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 1 39 with glycoprotein 220:231 arg1 role 249:252 a proposed role 238:252 a proposed role in bone mineralization 238:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 7 40 theme severe 1346:1351 arg1 TNSALP 1310:1315 TNSALP 1310:1315 TNSALP (N430S) 1310:1323 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 7 40 theme severe 1346:1351 arg1 allele 1353:1358 a severe allele 1344:1358 a severe allele without strong ALP activity 1344:1386 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 6 41 theme TNSALP 1063:1068 arg1 mutants 1078:1084 TNSALP (N430Q) and TNSALP (N430E) mutants 1044:1084 mutants 1078:1084 However, we found that TNSALP (N430Q) and TNSALP (N430E) mutants, but not a TNSALP (N430D) mutant, failed to form dimers. 26797772 6 41 theme TNSALP 1063:1068 arg1 N430E 1071:1075 N430E 1071:1075 N430E 1071:1075 However, we found that TNSALP (N430Q) and TNSALP (N430E) mutants, but not a TNSALP (N430D) mutant, failed to form dimers. 26797772 7 42 theme N430S 1155:1159 arg1 mutant 1162:1167 The TNSALP (N430S) mutant 1143:1167 The TNSALP (N430S) mutant linked to infantile HPP 1143:1191 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 7 42 theme N430S 1155:1159 arg1 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 2 43 theme genetic 370:376 arg1 disease 378:384 a genetic disease 368:384 a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity 368:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 43 theme genetic 370:376 arg1 hypophosphatasia 344:359 hypophosphatasia 344:359 hypophosphatasia (HPP) 344:365 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 5 44 contain has 745:747 arg1 TNSALP 738:743 TNSALP 738:743 TNSALP 738:743 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 5 44 contain has 745:747 arg2 N-glycans 754:762 five N-glycans 749:762 five N-glycans 749:762 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 7 45 theme TNSALP 1147:1152 arg1 mutant 1162:1167 The TNSALP (N430S) mutant 1143:1167 The TNSALP (N430S) mutant linked to infantile HPP 1143:1191 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 7 45 theme TNSALP 1147:1152 arg1 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 1 46 gly glycoprotein 220:231 arg1 glycoprotein 220:231 a membrane glycoprotein 209:231 a membrane glycoprotein with a proposed role in bone mineralization 209:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 46 gly glycoprotein 220:231 arg1 phosphatase 185:195 Tissue-nonspecific alkaline phosphatase 157:195 Tissue-nonspecific alkaline phosphatase (TNSALP) 157:204 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 8 47 theme wild-type 1526:1534 arg1 TNSALP 1536:1541 wild-type TNSALP 1526:1541 wild-type TNSALP 1526:1541 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 7 48 attach linked 1169:1174 arg2 mutant 1162:1167 The TNSALP (N430S) mutant 1143:1167 The TNSALP (N430S) mutant linked to infantile HPP 1143:1191 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 7 48 attach linked 1169:1174 arg2 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 glycosylation-defective 1197:1219 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 7 48 attach linked 1169:1174 arg1 HPP 1189:1191 infantile HPP 1179:1191 infantile HPP 1179:1191 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 9 49 theme minimal 1750:1756 arg1 requirement 1758:1768 the minimal requirement 1746:1768 the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell 1746:1864 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 49 theme minimal 1750:1756 arg1 N-glycans 1708:1716 three N-glycans 1702:1716 three N-glycans on N230, N271 and N303 1702:1739 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 1 50 theme membrane 211:218 arg1 glycoprotein 220:231 a membrane glycoprotein 209:231 a membrane glycoprotein with a proposed role in bone mineralization 209:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 50 theme membrane 211:218 arg1 phosphatase 185:195 Tissue-nonspecific alkaline phosphatase 157:195 Tissue-nonspecific alkaline phosphatase (TNSALP) 157:204 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 5 51 theme single 811:816 arg1 mutants 836:842 individual single N-glycan deletion mutants 800:842 individual single N-glycan deletion mutants of TNSALP 800:852 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 9 52 from N-glycans 1708:1716 arg1 N271 1727:1730 N271 1727:1730 N271 1727:1730 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 52 from N-glycans 1708:1716 arg1 N230 1721:1724 N230 1721:1724 N230 1721:1724 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 52 from N-glycans 1708:1716 arg1 N303 1736:1739 N303 1736:1739 N303 1736:1739 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 4 53 theme sites 616:620 arg1 N430 623:626 N430 623:626 N430 623:626 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 4 53 theme sites 616:620 arg1 one 603:605 one 603:605 one 603:605 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 4 53 theme sites 616:620 arg1 sites 616:620 these sites 610:620 these sites 610:620 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 5 54 theme deletion 827:834 arg1 mutants 836:842 individual single N-glycan deletion mutants 800:842 individual single N-glycan deletion mutants of TNSALP 800:852 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 3 55 contain has 487:489 arg2 sites 522:526 five potential N-glycosylation sites 491:526 five potential N-glycosylation sites 491:526 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 3 55 contain has 487:489 arg1 TNSALP 480:485 TNSALP 480:485 TNSALP 480:485 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 8 56 from present 1489:1495 arg1 cells 1558:1562 transfected cells 1546:1562 transfected cells 1546:1562 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 1 57 from role 249:252 arg1 mineralization 262:275 bone mineralization 257:275 bone mineralization 257:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 5 58 theme ALP 896:898 arg1 activity 900:907 ALP activity 896:907 ALP activity 896:907 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 9 59 theme TNSALP 1804:1809 arg1 function 1792:1799 function 1792:1799 function 1792:1799 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 9 59 theme TNSALP 1804:1809 arg1 structure 1778:1786 structure 1778:1786 structure 1778:1786 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 1 60 theme Tissue-nonspecific 157:174 arg1 glycoprotein 220:231 a membrane glycoprotein 209:231 a membrane glycoprotein with a proposed role in bone mineralization 209:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 60 theme Tissue-nonspecific 157:174 arg1 TNSALP 198:203 TNSALP 198:203 TNSALP 198:203 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 1 60 theme Tissue-nonspecific 157:174 arg1 phosphatase 185:195 Tissue-nonspecific alkaline phosphatase 157:195 Tissue-nonspecific alkaline phosphatase (TNSALP) 157:204 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 5 61 theme dimeric 865:871 arg1 structure 873:881 the dimeric structure 861:881 the dimeric structure required for ALP activity 861:907 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 7 62 theme similar 1245:1251 arg1 dimerise 1235:1242 dimerise 1235:1242 dimerise 1235:1242 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 4 63 from one 603:605 arg1 mutation 591:598 A mutation 589:598 A mutation at one of these sites, N430, 589:627 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 2 64 with patients 330:337 arg1 disease 378:384 a genetic disease 368:384 a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity 368:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 64 with patients 330:337 arg1 HPP 362:364 HPP 362:364 HPP 362:364 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 2 64 with patients 330:337 arg1 hypophosphatasia 344:359 hypophosphatasia 344:359 hypophosphatasia (HPP) 344:365 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 0 65 from mutations 24:32 arg1 phosphatase 65:75 tissue-nonspecific alkaline phosphatase 37:75 tissue-nonspecific alkaline phosphatase 37:75 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 9 66 theme stable 1838:1843 arg1 expression 1845:1854 its stable expression 1834:1854 its stable expression in a cell 1834:1864 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 5 67 theme single 945:950 arg1 N-glycan 952:959 any single N-glycan 941:959 any single N-glycan 941:959 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 0 68 theme infantile 129:137 arg1 hypophosphatasia 139:154 infantile hypophosphatasia 129:154 infantile hypophosphatasia 129:154 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 2 69 theme serum 460:464 arg1 activity 470:477 serum ALP activity 460:477 serum ALP activity 460:477 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 3 70 gly N-glycosylation 506:520 arg2 five 491:494 five 491:494 five 491:494 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 3 70 gly N-glycosylation 506:520 arg2 sites 522:526 five potential N-glycosylation sites 491:526 five potential N-glycosylation sites 491:526 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 0 71 theme tissue-nonspecific 37:54 arg1 phosphatase 65:75 tissue-nonspecific alkaline phosphatase 37:75 tissue-nonspecific alkaline phosphatase 37:75 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 8 72 attach present 1489:1495 arg1 cells 1558:1562 transfected cells 1546:1562 transfected cells 1546:1562 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 8 72 attach present 1489:1495 arg2 TNSALP 1449:1454 TNSALP 1449:1454 TNSALP devoid of all five N-glycans 1449:1483 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 9 73 theme mutants 1670:1676 arg1 series 1632:1637 a series 1630:1637 a series of multiple N-glycan depletion mutants in TNSALP 1630:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 26797772 8 74 theme single 1415:1420 arg1 mutants 1440:1446 individual single N-glycan deletion mutants 1404:1446 individual single N-glycan deletion mutants 1404:1446 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 1 75 theme proposed 240:247 arg1 role 249:252 a proposed role 238:252 a proposed role in bone mineralization 238:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 2 76 from mutations 286:294 arg1 TNSALP 299:304 TNSALP 299:304 TNSALP 299:304 Indeed, mutations in TNSALP have been identified in patients with hypophosphatasia (HPP), a genetic disease characterized by hypomineralization of bone and teeth and a deficiency in serum ALP activity. 26797772 7 77 theme infantile 1179:1187 arg1 HPP 1189:1191 infantile HPP 1179:1191 infantile HPP 1179:1191 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 8 78 theme deletion 1431:1438 arg1 mutants 1440:1446 individual single N-glycan deletion mutants 1404:1446 individual single N-glycan deletion mutants 1404:1446 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 8 79 from cells 1558:1562 arg1 present 1489:1495 present 1489:1495 present 1489:1495 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 5 80 theme transfected 767:777 arg1 cells 785:789 transfected COS-1 cells 767:789 transfected COS-1 cells 767:789 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 4 81 located detected 642:649 arg2 mutation 591:598 A mutation 589:598 A mutation at one of these sites, N430, 589:627 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 4 81 located detected 642:649 arg1 patient 656:662 a patient 654:662 a patient with infantile HPP 654:681 A mutation at one of these sites, N430, was recently detected in a patient with infantile HPP. 26797772 7 82 theme strong 1368:1373 arg1 activity 1379:1386 strong ALP activity 1368:1386 strong ALP activity 1368:1386 The TNSALP (N430S) mutant linked to infantile HPP was glycosylation-defective and unable to dimerise, similar to TNSALP (N430Q) and TNSALP (N430E) mutants; therefore, TNSALP (N430S) was established as a severe allele without strong ALP activity. 26797772 5 83 theme vital 969:973 arg1 role 975:978 a vital role 967:978 a vital role 967:978 Using site-directed mutagenesis, we demonstrated that TNSALP has five N-glycans in transfected COS-1 cells and that individual single N-glycan deletion mutants of TNSALP retain the dimeric structure required for ALP activity, excluding the possibility that any single N-glycan plays a vital role in the structure and function of TNSALP. 26797772 8 84 theme devoid 1456:1461 arg1 TNSALP 1449:1454 TNSALP 1449:1454 TNSALP devoid of all five N-glycans 1449:1483 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 3 85 theme potential 496:504 arg1 sites 522:526 five potential N-glycosylation sites 491:526 five potential N-glycosylation sites 491:526 TNSALP has five potential N-glycosylation sites at N140, N230, N271, N303 and N430 by standard nomenclature. 26797772 8 86 theme lesser 1507:1512 arg1 extent 1514:1519 a much lesser extent 1500:1519 a much lesser extent than wild-type TNSALP 1500:1541 By contrast to individual single N-glycan deletion mutants, TNSALP devoid of all five N-glycans was present to a much lesser extent than wild-type TNSALP in transfected cells, possibly reflecting its instability. 26797772 1 87 theme bone 257:260 arg1 mineralization 262:275 bone mineralization 257:275 bone mineralization 257:275 Tissue-nonspecific alkaline phosphatase (TNSALP) is a membrane glycoprotein with a proposed role in bone mineralization. 26797772 0 88 attach linked 119:124 arg1 hypophosphatasia 139:154 infantile hypophosphatasia 129:154 infantile hypophosphatasia 129:154 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 0 88 attach linked 119:124 arg2 mutations 24:32 Glycosylation-deficient mutations 0:32 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase 0:75 Glycosylation-deficient mutations in tissue-nonspecific alkaline phosphatase impair its structure and function and are linked to infantile hypophosphatasia. 26797772 9 89 theme multiple 1642:1649 arg1 mutants 1670:1676 multiple N-glycan depletion mutants 1642:1676 multiple N-glycan depletion mutants in TNSALP 1642:1686 A comprehensive analysis of a series of multiple N-glycan depletion mutants in TNSALP revealed that three N-glycans on N230, N271 and N303 were the minimal requirement for the structure and function of TNSALP and a prerequisite for its stable expression in a cell. 28920453 6 0 theme other 942:946 arg1 biotherapeutics 948:962 other biotherapeutics 942:962 not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations 917:996 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 1 1 theme recombinant 156:166 arg1 rhEPOs 191:196 rhEPOs 191:196 rhEPOs 191:196 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 1 1 theme recombinant 156:166 arg1 erythropoietins 174:188 recombinant human erythropoietins 156:188 recombinant human erythropoietins (rhEPOs) 156:197 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 0 2 theme case 78:81 arg1 study 83:87 a case study 76:87 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 1 3 theme human 168:172 arg1 rhEPOs 191:196 rhEPOs 191:196 rhEPOs 191:196 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 1 3 theme human 168:172 arg1 erythropoietins 174:188 recombinant human erythropoietins 156:188 recombinant human erythropoietins (rhEPOs) 156:197 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 0 4 theme novel 92:96 arg1 protein 125:131 novel erythropoiesis stimulating protein 92:131 novel erythropoiesis stimulating protein 92:131 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 6 5 theme tandem 839:844 arg1 CONCLUSION 850:859 tandem MS. CONCLUSION 839:859 tandem MS. CONCLUSION 839:859 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 3 6 with O-glycans 528:536 arg1 sensitivity 548:558 high sensitivity 543:558 high sensitivity 543:558 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 2 7 theme glycosylation 300:312 arg1 characterization 280:295 comprehensive characterization 266:295 comprehensive characterization of glycosylation 266:312 Thus, comprehensive characterization of glycosylation is vital to assess the biotherapeutic quality and establish the equivalency of biosimilar rhEPOs. 28920453 4 8 theme native 616:621 arg1 O-glycans 623:631 profile native O-glycans 608:631 profile native O-glycans on rhEPOs 608:641 We developed selective and sensitive method to profile native O-glycans on rhEPOs. 28920453 6 9 theme rhEPOs 926:931 arg1 QC 892:893 QC 892:893 QC 892:893 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 6 9 theme rhEPOs 926:931 arg1 analysis 905:912 batch analysis 899:912 batch analysis 899:912 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 4 10 theme profile 608:614 arg1 O-glycans 623:631 profile native O-glycans 608:631 profile native O-glycans on rhEPOs 608:641 We developed selective and sensitive method to profile native O-glycans on rhEPOs. 28920453 0 11 theme stimulating 113:123 arg1 protein 125:131 novel erythropoiesis stimulating protein 92:131 novel erythropoiesis stimulating protein 92:131 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 3 12 theme high 543:546 arg1 sensitivity 548:558 high sensitivity 543:558 high sensitivity 543:558 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 1 13 theme erythropoietins 174:188 arg1 Glycosylation 139:151 Glycosylation 139:151 Glycosylation of recombinant human erythropoietins (rhEPOs) 139:197 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 0 14 theme erythropoiesis 98:111 arg1 protein 125:131 novel erythropoiesis stimulating protein 92:131 novel erythropoiesis stimulating protein 92:131 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 0 15 theme O-glycosylation 40:54 arg1 analysis 28:35 Sensitive and comprehensive analysis 0:35 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 6 16 theme MS. 846:848 arg1 CONCLUSION 850:859 tandem MS. CONCLUSION 839:859 tandem MS. CONCLUSION 839:859 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 2 17 theme comprehensive 266:278 arg1 characterization 280:295 comprehensive characterization 266:295 comprehensive characterization of glycosylation 266:312 Thus, comprehensive characterization of glycosylation is vital to assess the biotherapeutic quality and establish the equivalency of biosimilar rhEPOs. 28920453 6 18 theme biotherapeutics 948:962 arg1 QC 892:893 QC 892:893 QC 892:893 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 6 18 theme biotherapeutics 948:962 arg1 analysis 905:912 batch analysis 899:912 batch analysis 899:912 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 4 19 from O-glycans 623:631 arg1 rhEPOs 636:641 rhEPOs 636:641 rhEPOs 636:641 We developed selective and sensitive method to profile native O-glycans on rhEPOs. 28920453 5 20 from O-glycosylation 653:667 arg1 rhEPO 672:676 rhEPO 672:676 rhEPO including O-acetylation on a sialic acid 672:717 RESULTS: O-glycosylation on rhEPO including O-acetylation on a sialic acid was comprehensively characterized. 28920453 5 21 theme sialic 707:712 arg1 acid 714:717 a sialic acid 705:717 a sialic acid 705:717 RESULTS: O-glycosylation on rhEPO including O-acetylation on a sialic acid was comprehensively characterized. 28920453 2 22 theme rhEPOs 404:409 arg1 equivalency 378:388 the equivalency 374:388 the equivalency of biosimilar rhEPOs 374:409 Thus, comprehensive characterization of glycosylation is vital to assess the biotherapeutic quality and establish the equivalency of biosimilar rhEPOs. 28920453 0 23 theme Sensitive 0:8 arg1 analysis 28:35 Sensitive and comprehensive analysis 0:35 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 4 24 theme selective 574:582 arg1 method 598:603 selective and sensitive method 574:603 selective and sensitive method 574:603 We developed selective and sensitive method to profile native O-glycans on rhEPOs. 28920453 2 25 theme biosimilar 393:402 arg1 rhEPOs 404:409 biosimilar rhEPOs 393:409 biosimilar rhEPOs 393:409 Thus, comprehensive characterization of glycosylation is vital to assess the biotherapeutic quality and establish the equivalency of biosimilar rhEPOs. 28920453 0 26 theme comprehensive 14:26 arg1 analysis 28:35 Sensitive and comprehensive analysis 0:35 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 4 27 theme sensitive 588:596 arg1 method 598:603 selective and sensitive method 574:603 selective and sensitive method 574:603 We developed selective and sensitive method to profile native O-glycans on rhEPOs. 28920453 0 28 theme protein 125:131 arg1 study 83:87 a case study 76:87 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 3 29 theme glycan 429:434 arg1 analysis 436:443 current glycan analysis 421:443 current glycan analysis 421:443 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 6 30 theme batch 899:903 arg1 analysis 905:912 batch analysis 899:912 batch analysis 899:912 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 5 31 gly O-glycosylation 653:667 arg1 rhEPO 672:676 rhEPO 672:676 rhEPO including O-acetylation on a sialic acid 672:717 RESULTS: O-glycosylation on rhEPO including O-acetylation on a sialic acid was comprehensively characterized. 28920453 3 32 theme analytical 499:508 arg1 tools 510:514 analytical tools 499:514 analytical tools 499:514 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 3 33 theme current 421:427 arg1 analysis 436:443 current glycan analysis 421:443 current glycan analysis 421:443 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 5 34 from rhEPO 672:676 arg1 acid 714:717 a sialic acid 705:717 a sialic acid 705:717 RESULTS: O-glycosylation on rhEPO including O-acetylation on a sialic acid was comprehensively characterized. 28920453 1 35 gly Glycosylation 139:151 arg1 rhEPOs 191:196 rhEPOs 191:196 rhEPOs 191:196 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 1 35 gly Glycosylation 139:151 arg1 erythropoietins 174:188 recombinant human erythropoietins 156:188 recombinant human erythropoietins (rhEPOs) 156:197 AIM: Glycosylation of recombinant human erythropoietins (rhEPOs) is significantly associated with drug's quality and potency. 28920453 6 36 theme O-acetyl-modification 793:813 arg1 site 815:818 O-acetyl-modification site 793:818 O-acetyl-modification site 793:818 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 3 37 theme tools 510:514 arg1 absence 488:494 the absence 484:494 the absence of analytical tools to liberate O-glycans with high sensitivity 484:558 However, current glycan analysis mainly focuses on the N-glycans due to the absence of analytical tools to liberate O-glycans with high sensitivity. 28920453 0 38 dep analysis 28:35 arg1 study 83:87 a case study 76:87 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 0:132 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28920453 2 39 theme biotherapeutic 337:350 arg1 quality 352:358 the biotherapeutic quality 333:358 the biotherapeutic quality 333:358 Thus, comprehensive characterization of glycosylation is vital to assess the biotherapeutic quality and establish the equivalency of biosimilar rhEPOs. 28920453 6 40 theme O-glycan 770:777 arg1 structure 779:787 O-glycan structure 770:787 O-glycan structure 770:787 Details such as O-glycan structure and O-acetyl-modification site were obtained from tandem MS. CONCLUSION: This method may be applied to QC and batch analysis of not only rhEPOs but also other biotherapeutics bearing multiple O-glycosylations. 28920453 0 41 from analysis 28:35 arg1 biotherapeutics 59:73 biotherapeutics 59:73 biotherapeutics 59:73 Sensitive and comprehensive analysis of O-glycosylation in biotherapeutics: a case study of novel erythropoiesis stimulating protein. 28955811 0 0 theme enzymatic 67:75 arg1 activity 77:84 enzymatic activity 67:84 enzymatic activity 67:84 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 28955811 7 1 theme decreased 1229:1237 arg1 levels 1239:1244 decreased levels 1229:1244 decreased levels of secreted active hCES2 1229:1269 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 4 2 theme enzymatic 803:811 arg1 deglycosylation 813:827 enzymatic deglycosylation 803:827 three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites 781:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 3 dep site 911:914 arg1 ii 907:908 ii 907:908 ii 907:908 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 3 dep site 911:914 arg1 mutagenesis 925:935 directed mutagenesis 916:935 directed mutagenesis of each 916:943 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 3 4 theme enzyme 640:645 arg1 forms 627:631 alternative active forms 608:631 alternative active forms of the enzyme that might be adequate for structure elucidation 608:694 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 2 5 theme glycosylation 316:328 arg1 heterogeneity 330:342 glycosylation heterogeneity 316:342 glycosylation heterogeneity 316:342 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 4 6 dep incubation 864:873 arg1 ii 860:861 ii 860:861 ii 860:861 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 7 7 gly N-glycosylation 1200:1214 arg2 sites 1216:1220 N-glycosylation sites 1200:1220 N-glycosylation sites 1200:1220 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 8 8 theme mutants 1322:1328 arg1 thermostability 1285:1299 the thermostability 1281:1299 the thermostability of the glycosylation mutants 1281:1328 However, the thermostability of the glycosylation mutants was decreased. 28955811 2 9 theme inhibitors 481:490 arg1 development 447:457 the development 443:457 the development of new substrates and inhibitors 443:490 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 3 10 theme structure 674:682 arg1 elucidation 684:694 structure elucidation 674:694 structure elucidation 674:694 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 9 11 theme hCES2 1508:1512 arg1 activity 1487:1494 the activity 1483:1494 the activity of secreted hCES2 1483:1512 The results indicated that glycans are involved, to some extent in protein folding in vivo, however, removal of glycans does not abrogate the activity of secreted hCES2. 28955811 4 12 with incubation 864:873 arg1 tunicamycin 894:904 the inhibitor tunicamycin 880:904 the inhibitor tunicamycin 880:904 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 7 13 dep each 1186:1189 arg1 sites 1216:1220 N-glycosylation sites 1200:1220 N-glycosylation sites 1200:1220 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 7 14 theme each 1186:1189 arg1 mutation 1174:1181 mutation 1174:1181 mutation of each and both N-glycosylation sites 1174:1220 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 4 15 theme secreted 736:743 arg1 form 745:748 a secreted form 734:748 a secreted form of hCES2 734:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 7 16 theme secreted 1249:1256 arg1 hCES2 1265:1269 secreted active hCES2 1249:1269 secreted active hCES2 1249:1269 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 2 17 theme substrates 466:475 arg1 development 447:457 the development 443:457 the development of new substrates and inhibitors 443:490 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 5 18 theme enzyme 1037:1042 arg1 activity 1044:1051 enzyme activity 1037:1051 enzyme activity 1037:1051 Deglycosylated protein did not show a detectable decrease in enzyme activity. 28955811 3 19 theme present 497:503 arg1 work 505:508 The present work 493:508 The present work 493:508 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 4 20 dep approaches 787:796 arg1 incubation 864:873 (ii) incubation 859:873 three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites 781:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 20 dep approaches 787:796 arg1 deglycosylation 813:827 enzymatic deglycosylation 803:827 three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites 781:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 20 dep approaches 787:796 arg1 sites 969:973 both N-glycosylation sites 948:973 both N-glycosylation sites 948:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 20 dep approaches 787:796 arg1 site 911:914 ii) site directed mutagenesis of each or both N-glycosylation sites 907:973 site 911:914 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 9 21 theme secreted 1499:1506 arg1 hCES2 1508:1512 secreted hCES2 1499:1512 secreted hCES2 1499:1512 The results indicated that glycans are involved, to some extent in protein folding in vivo, however, removal of glycans does not abrogate the activity of secreted hCES2. 28955811 0 22 from Studies 26:32 arg1 role 41:44 the role 37:44 the role of glycosylation for enzymatic activity 37:84 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 28955811 7 23 theme both 1195:1198 arg1 mutation 1174:1181 mutation 1174:1181 mutation of each and both N-glycosylation sites 1174:1220 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 0 24 theme glycosylation 49:61 arg1 role 41:44 the role 37:44 the role of glycosylation for enzymatic activity 37:84 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 28955811 4 25 theme Partial 697:703 arg1 forms 725:729 Partial or non-glycosylated forms 697:729 Partial or non-glycosylated forms of a secreted form of hCES2 697:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 0 26 theme Human 0:4 arg1 carboxylesterase 6:21 Human carboxylesterase 2 0:23 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 0:85 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 28955811 6 27 theme hCES2 1121:1125 arg1 levels 1102:1107 decreased levels 1092:1107 decreased levels of secreted hCES2 1092:1125 On the other hand, tunicamycin led to decreased levels of secreted hCES2 but the enzyme was still active. 28955811 3 28 theme glycans 539:545 arg1 involvement 524:534 the involvement 520:534 the involvement of glycans in hCES2 activity and thermo stability 520:584 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 3 29 theme alternative 608:618 arg1 forms 627:631 alternative active forms 608:631 alternative active forms of the enzyme that might be adequate for structure elucidation 608:694 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 5 30 from decrease 1025:1032 arg1 activity 1044:1051 enzyme activity 1037:1051 enzyme activity 1037:1051 Deglycosylated protein did not show a detectable decrease in enzyme activity. 28955811 0 31 dep carboxylesterase 6:21 arg1 Studies 26:32 Studies 26:32 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 0:85 Human carboxylesterase 2: Studies on the role of glycosylation for enzymatic activity. 28955811 7 32 theme active 1258:1263 arg1 hCES2 1265:1269 secreted active hCES2 1249:1269 secreted active hCES2 1249:1269 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 1 33 theme Human 87:91 arg1 hCES2 113:117 hCES2 113:117 hCES2 113:117 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 33 theme Human 87:91 arg1 glycoprotein 125:136 a glycoprotein 123:136 a glycoprotein 123:136 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 33 theme Human 87:91 arg1 carboxylesterase 93:108 Human carboxylesterase 2 87:110 Human carboxylesterase 2 (hCES2) 87:118 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 34 theme drugs 168:172 arg1 metabolism 154:163 the metabolism 150:163 the metabolism of drugs and several environmental xenobiotics 150:210 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 2 35 theme hCES2 398:402 arg1 structure 385:393 the 3D structure 378:393 the 3D structure of hCES2 378:402 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 1 36 theme challenging 259:269 arg1 task 271:274 a challenging task 257:274 a challenging task 257:274 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 36 theme challenging 259:269 arg1 crystallization 219:233 crystallization 219:233 crystallization 219:233 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 6 37 theme decreased 1092:1100 arg1 levels 1102:1107 decreased levels 1092:1107 decreased levels of secreted hCES2 1092:1125 On the other hand, tunicamycin led to decreased levels of secreted hCES2 but the enzyme was still active. 28955811 4 38 theme non-glycosylated 708:723 arg1 forms 725:729 Partial or non-glycosylated forms 697:729 Partial or non-glycosylated forms of a secreted form of hCES2 697:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 2 39 theme structure 385:393 arg1 disclosure 364:373 the disclosure 360:373 the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors 360:490 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 9 40 theme glycans 1457:1463 arg1 removal 1446:1452 removal 1446:1452 removal of glycans 1446:1463 The results indicated that glycans are involved, to some extent in protein folding in vivo, however, removal of glycans does not abrogate the activity of secreted hCES2. 28955811 1 41 theme several 178:184 arg1 xenobiotics 200:210 several environmental xenobiotics 178:210 several environmental xenobiotics 178:210 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 6 42 theme secreted 1112:1119 arg1 hCES2 1121:1125 secreted hCES2 1112:1125 secreted hCES2 1112:1125 On the other hand, tunicamycin led to decreased levels of secreted hCES2 but the enzyme was still active. 28955811 2 43 theme 3D 382:383 arg1 structure 385:393 the 3D structure 378:393 the 3D structure of hCES2 378:402 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 4 44 theme directed 916:923 arg1 mutagenesis 925:935 directed mutagenesis 916:935 directed mutagenesis of each 916:943 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 45 dep deglycosylation 813:827 arg1 i 800:800 i 800:800 i 800:800 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 9 46 from extent 1402:1407 arg1 folding 1420:1426 protein folding 1412:1426 protein folding in vivo 1412:1434 The results indicated that glycans are involved, to some extent in protein folding in vivo, however, removal of glycans does not abrogate the activity of secreted hCES2. 28955811 1 47 theme environmental 186:198 arg1 xenobiotics 200:210 several environmental xenobiotics 178:210 several environmental xenobiotics 178:210 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 2 48 theme upmost 422:427 arg1 relevance 429:437 upmost relevance 422:437 upmost relevance 422:437 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 4 49 theme N-glycosidase 842:854 arg1 F 856:856 peptide N-glycosidase F 834:856 peptide N-glycosidase F 834:856 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 5 50 theme detectable 1014:1023 arg1 decrease 1025:1032 a detectable decrease 1012:1032 a detectable decrease in enzyme activity 1012:1051 Deglycosylated protein did not show a detectable decrease in enzyme activity. 28955811 4 51 gly N-glycosylation 953:967 arg2 sites 969:973 both N-glycosylation sites 948:973 both N-glycosylation sites 948:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 6 52 theme other 1061:1065 arg1 hand 1067:1070 the other hand 1057:1070 the other hand 1057:1070 On the other hand, tunicamycin led to decreased levels of secreted hCES2 but the enzyme was still active. 28955811 4 53 theme peptide 834:840 arg1 F 856:856 peptide N-glycosidase F 834:856 peptide N-glycosidase F 834:856 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 3 54 theme hCES2 550:554 arg1 activity 556:563 hCES2 activity 550:563 hCES2 activity 550:563 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 4 55 theme hCES2 753:757 arg1 form 745:748 a secreted form 734:748 a secreted form of hCES2 734:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 5 56 theme Deglycosylated 976:989 arg1 protein 991:997 Deglycosylated protein 976:997 Deglycosylated protein 976:997 Deglycosylated protein did not show a detectable decrease in enzyme activity. 28955811 7 57 theme hCES2 1265:1269 arg1 levels 1239:1244 decreased levels 1229:1244 decreased levels of secreted active hCES2 1229:1269 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 3 58 from involvement 524:534 arg1 activity 556:563 hCES2 activity 550:563 hCES2 activity 550:563 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 3 58 from involvement 524:534 arg1 stability 576:584 thermo stability 569:584 thermo stability 569:584 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 1 59 theme xenobiotics 200:210 arg1 metabolism 154:163 the metabolism 150:163 the metabolism of drugs and several environmental xenobiotics 150:210 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 60 gly glycoprotein 125:136 arg1 glycoprotein 125:136 a glycoprotein 123:136 a glycoprotein 123:136 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 1 60 gly glycoprotein 125:136 arg1 carboxylesterase 93:108 Human carboxylesterase 2 87:110 Human carboxylesterase 2 (hCES2) 87:118 Human carboxylesterase 2 (hCES2) is a glycoprotein involved in the metabolism of drugs and several environmental xenobiotics, whose crystallization has been proved to be a challenging task. 28955811 3 61 theme active 620:625 arg1 forms 627:631 alternative active forms 608:631 alternative active forms of the enzyme that might be adequate for structure elucidation 608:694 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 4 62 theme form 745:748 arg1 forms 725:729 Partial or non-glycosylated forms 697:729 Partial or non-glycosylated forms of a secreted form of hCES2 697:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 9 63 theme protein 1412:1418 arg1 folding 1420:1426 protein folding 1412:1426 protein folding in vivo 1412:1434 The results indicated that glycans are involved, to some extent in protein folding in vivo, however, removal of glycans does not abrogate the activity of secreted hCES2. 28955811 2 64 theme new 462:464 arg1 substrates 466:475 new substrates 462:475 new substrates 462:475 This limitation could partly be due to glycosylation heterogeneity and has delayed the disclosure of the 3D structure of hCES2 which would be of upmost relevance for the development of new substrates and inhibitors. 28955811 7 65 theme N-glycosylation 1200:1214 arg1 sites 1216:1220 N-glycosylation sites 1200:1220 N-glycosylation sites 1200:1220 In agreement, mutation of each and both N-glycosylation sites led to decreased levels of secreted active hCES2. 28955811 3 66 theme thermo 569:574 arg1 stability 576:584 thermo stability 569:584 thermo stability 569:584 The present work evaluated the involvement of glycans in hCES2 activity and thermo stability in an attempt to find alternative active forms of the enzyme that might be adequate for structure elucidation. 28955811 4 67 gly non-glycosylated 708:723 arg1 form 745:748 a secreted form 734:748 a secreted form of hCES2 734:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 67 gly non-glycosylated 708:723 arg1 forms 725:729 Partial or non-glycosylated forms 697:729 Partial or non-glycosylated forms of a secreted form of hCES2 697:757 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 8 68 theme glycosylation 1308:1320 arg1 mutants 1322:1328 the glycosylation mutants 1304:1328 the glycosylation mutants 1304:1328 However, the thermostability of the glycosylation mutants was decreased. 28955811 4 69 with deglycosylation 813:827 arg1 F 856:856 peptide N-glycosidase F 834:856 peptide N-glycosidase F 834:856 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 70 theme each 940:943 arg1 mutagenesis 925:935 directed mutagenesis 916:935 directed mutagenesis of each 916:943 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 71 theme N-glycosylation 953:967 arg1 sites 969:973 both N-glycosylation sites 948:973 both N-glycosylation sites 948:973 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 4 72 theme inhibitor 884:892 arg1 tunicamycin 894:904 the inhibitor tunicamycin 880:904 the inhibitor tunicamycin 880:904 Partial or non-glycosylated forms of a secreted form of hCES2 have been obtained by three approaches: (i) enzymatic deglycosylation with peptide N-glycosidase F; (ii) incubation with the inhibitor tunicamycin; ii) site directed mutagenesis of each or both N-glycosylation sites. 28955811 5 73 gly Deglycosylated 976:989 arg1 protein 991:997 Deglycosylated protein 976:997 Deglycosylated protein 976:997 Deglycosylated protein did not show a detectable decrease in enzyme activity. 25646460 4 0 from impact 682:687 arg1 activity 707:714 disease-homing activity 692:714 disease-homing activity 692:714 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 4 1 theme characterization 774:789 arg1 importance 734:743 the importance 730:743 the importance of glycosylation control and characterization during process development 730:816 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 3 2 theme microheterogeneity 560:577 arg1 view 541:544 a detailed view 530:544 a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein 530:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 2 3 with studies 331:337 arg1 antibody 378:385 a novel IL9-armed cancer-specific antibody 344:385 a novel IL9-armed cancer-specific antibody 344:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 3 4 theme present 579:585 arg1 microheterogeneity 560:577 the glycan microheterogeneity 549:577 the glycan microheterogeneity present on the IL9 portion of the recombinant protein 549:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 4 5 theme glycan 648:653 arg1 structures 655:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 2 6 theme detailed 306:313 arg1 studies 331:337 detailed biodistribution studies 306:337 detailed biodistribution studies with a novel IL9-armed cancer-specific antibody 306:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 3 7 theme protein 625:631 arg1 portion 598:604 the IL9 portion 590:604 the IL9 portion of the recombinant protein 590:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 1 8 theme endothelial 272:282 arg1 lining 284:289 the endothelial lining 268:289 the endothelial lining 268:289 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 3 9 theme recombinant 613:623 arg1 protein 625:631 the recombinant protein 609:631 the recombinant protein 609:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 4 10 theme process 798:804 arg1 development 806:816 process development 798:816 process development 798:816 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 2 11 theme novel 346:350 arg1 antibody 378:385 a novel IL9-armed cancer-specific antibody 344:385 a novel IL9-armed cancer-specific antibody 344:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 3 12 theme IL9 594:596 arg1 portion 598:604 the IL9 portion 590:604 the IL9 portion of the recombinant protein 590:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 1 13 theme therapeutic 179:189 arg1 activity 191:198 therapeutic activity 179:198 therapeutic activity 179:198 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 3 14 theme glycan 505:510 arg1 analysis 512:519 Site-specific glycan analysis 491:519 Site-specific glycan analysis 491:519 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 0 15 theme Glycosylation 0:12 arg1 profiles 14:21 Glycosylation profiles 0:21 Glycosylation profiles 0:21 Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies. 25646460 2 16 theme clear 404:408 arg1 correlation 410:420 a clear correlation 402:420 a clear correlation between N-linked glycan structures and tumor-targeting efficiencies 402:488 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 1 17 dep vessels 155:161 arg1 out 142:144 out 142:144 out 142:144 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 2 18 theme tumor-targeting 461:475 arg1 efficiencies 477:488 tumor-targeting efficiencies 461:488 tumor-targeting efficiencies 461:488 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 4 19 theme Nonsialylated 634:646 arg1 structures 655:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 2 20 theme biodistribution 315:329 arg1 studies 331:337 detailed biodistribution studies 306:337 detailed biodistribution studies with a novel IL9-armed cancer-specific antibody 306:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 3 21 theme detailed 532:539 arg1 view 541:544 a detailed view 530:544 a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein 530:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 0 22 theme disease-targeting 51:67 arg1 properties 69:78 disease-targeting properties 51:78 disease-targeting properties 51:78 Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies. 25646460 3 23 theme glycan 553:558 arg1 microheterogeneity 560:577 the glycan microheterogeneity 549:577 the glycan microheterogeneity present on the IL9 portion of the recombinant protein 549:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 4 24 contain have 666:669 arg2 impact 682:687 a negative impact 671:687 a negative impact on disease-homing activity 671:714 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 4 24 contain have 666:669 arg1 structures 655:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 4 25 theme glycosylation 748:760 arg1 importance 734:743 the importance 730:743 the importance of glycosylation control and characterization during process development 730:816 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 2 26 theme N-linked 430:437 arg1 structures 446:455 N-linked glycan structures 430:455 N-linked glycan structures 430:455 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 2 27 link N-linked 430:437 arg1 structures 446:455 N-linked glycan structures 430:455 N-linked glycan structures 430:455 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 2 28 theme glycan 439:444 arg1 structures 446:455 N-linked glycan structures 430:455 N-linked glycan structures 430:455 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 3 29 theme Site-specific 491:503 arg1 analysis 512:519 Site-specific glycan analysis 491:519 Site-specific glycan analysis 491:519 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 3 30 attach present 579:585 arg2 microheterogeneity 560:577 the glycan microheterogeneity 549:577 the glycan microheterogeneity present on the IL9 portion of the recombinant protein 549:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 3 30 attach present 579:585 arg1 portion 598:604 the IL9 portion 590:604 the IL9 portion of the recombinant protein 590:631 Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. 25646460 2 31 theme cancer-specific 362:376 arg1 antibody 378:385 a novel IL9-armed cancer-specific antibody 344:385 a novel IL9-armed cancer-specific antibody 344:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 4 32 gly Nonsialylated 634:646 arg1 structures 655:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures 634:664 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 2 33 theme IL9-armed 352:360 arg1 antibody 378:385 a novel IL9-armed cancer-specific antibody 344:385 a novel IL9-armed cancer-specific antibody 344:385 By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. 25646460 1 34 theme antibodies 116:125 arg1 critical 166:173 critical 166:173 critical 166:173 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 1 34 theme antibodies 116:125 arg1 ability 105:111 The ability 101:111 The ability of antibodies to extravasate out of blood vessels 101:161 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 4 35 dep glycosylation 748:760 arg1 control 762:768 control 762:768 control 762:768 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 1 36 theme blood 149:153 arg1 vessels 155:161 blood vessels 149:161 blood vessels 149:161 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 4 37 theme negative 673:680 arg1 impact 682:687 a negative impact 671:687 a negative impact on disease-homing activity 671:714 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 0 38 theme antibodies 89:98 arg1 extravasation 33:45 extravasation 33:45 extravasation 33:45 Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies. 25646460 0 38 theme antibodies 89:98 arg1 properties 69:78 disease-targeting properties 51:78 disease-targeting properties 51:78 Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies. 25646460 1 39 theme most 231:234 arg1 diseases 236:243 most diseases 231:243 most diseases 231:243 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 1 40 theme molecular 209:217 arg1 targets 219:225 molecular targets 209:225 molecular targets for most diseases 209:243 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 1 41 theme vessels 155:161 arg1 critical 166:173 critical 166:173 critical 166:173 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 1 41 theme vessels 155:161 arg1 ability 105:111 The ability 101:111 The ability of antibodies to extravasate out of blood vessels 101:161 The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. 25646460 4 42 theme disease-homing 692:705 arg1 activity 707:714 disease-homing activity 692:714 disease-homing activity 692:714 Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development. 25646460 0 43 theme armed 83:87 arg1 antibodies 89:98 armed antibodies 83:98 armed antibodies 83:98 Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies. 26537799 6 0 theme N-glycosylation 1117:1131 arg1 patterns 1133:1140 the N-glycosylation patterns 1113:1140 the N-glycosylation patterns of CRC cell lines 1113:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 4 1 theme CRC 570:572 arg1 lines 579:583 25 different CRC cell lines 557:583 25 different CRC cell lines 557:583 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 2 2 theme glyco 329:333 arg1 changes 346:352 these (glyco)biological changes 322:352 these (glyco)biological changes 322:352 CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. 26537799 4 3 theme lines 579:583 arg1 profiles 545:552 the N-glycan profiles 532:552 the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites 532:633 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 2 4 theme CRC 278:280 arg1 lines 287:291 CRC cell lines 278:291 CRC cell lines 278:291 CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. 26537799 1 5 theme protein 255:261 arg1 glycosylation 263:275 protein glycosylation 255:275 protein glycosylation 255:275 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 0 6 theme N-glycosylation 0:14 arg1 Profiling 16:24 N-glycosylation Profiling 0:24 N-glycosylation Profiling of Colorectal Cancer Cell Lines 0:56 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 1 7 theme Various 172:178 arg1 cancers 180:186 Various cancers 172:186 Various cancers such as colorectal cancer (CRC) 172:218 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 1 7 theme Various 172:178 arg1 cancer 207:212 colorectal cancer 196:212 colorectal cancer (CRC) 196:218 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 10 8 dep glycans 1906:1912 arg1 expression 1942:1951 villin mRNA expression 1930:1951 villin mRNA expression 1930:1951 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 10 8 dep glycans 1906:1912 arg1 CDX1 1918:1921 CDX1 1918:1921 CDX1 1918:1921 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 9 9 theme glycans 1745:1751 arg1 analysis 1733:1740 the structural analysis 1718:1740 the structural analysis of glycans 1718:1751 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 0 10 theme Homebox 131:137 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 10 11 theme cancer 1820:1825 arg1 markers 1832:1838 cancer cell markers 1820:1838 cancer cell markers 1820:1838 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 5 12 theme enzymatic 831:839 arg1 release 848:854 an optimized, high-throughput membrane-based enzymatic glycan release 786:854 release 848:854 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 6 13 theme α2,6-linked 965:975 arg1 acids 996:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 1 14 from alterations 240:250 arg1 glycosylation 263:275 protein glycosylation 255:275 protein glycosylation 255:275 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 4 15 attach derived 586:592 arg1 tumors 607:612 primary tumors 599:612 primary tumors 599:612 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 4 15 attach derived 586:592 arg1 sites 629:633 metastatic sites 618:633 metastatic sites 618:633 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 4 15 attach derived 586:592 arg2 lines 579:583 25 different CRC cell lines 557:583 25 different CRC cell lines 557:583 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 6 16 theme α2,3- 955:959 arg1 acids 996:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 8 17 theme intracellular 1481:1493 arg1 precursors 1495:1504 intracellular precursors 1481:1504 intracellular precursors 1481:1504 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 4 18 theme tumor 695:699 arg1 systems 707:713 glycobiological tumor model systems 679:713 glycobiological tumor model systems 679:713 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 0 19 theme mRNA 155:158 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 10 20 theme fucosylated 1894:1904 arg1 glycans 1906:1912 highly fucosylated glycans and CDX1 and/or villin mRNA expression 1887:1951 highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation 1887:1997 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 7 21 theme cancer 1379:1384 arg1 system 1392:1397 a glycobiological cancer model system 1361:1397 a glycobiological cancer model system 1361:1397 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 7 21 theme cancer 1379:1384 arg1 use 1354:1356 their use 1348:1356 their use 1348:1356 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 11 22 theme CRC-associated 2049:2062 arg1 changes 2071:2077 CRC-associated glycan changes 2049:2077 CRC-associated glycan changes 2049:2077 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 0 23 theme Fucosylation 81:92 arg1 Association 66:76 Association 66:76 Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 66:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 9 24 theme CRC 1562:1564 arg1 features 1585:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 6 25 theme N-glycosylation 1015:1029 arg1 analysis 1031:1038 N-glycosylation analysis 1015:1038 N-glycosylation analysis 1015:1038 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 8 26 theme cell 1535:1538 arg1 surface 1540:1546 the cell surface 1531:1546 the cell surface 1531:1546 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 10 27 theme features 1806:1813 arg1 correlation 1763:1773 correlation 1763:1773 correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes 1763:1853 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 9 28 theme line 1571:1574 arg1 features 1585:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 4 29 theme line 758:761 arg1 phenotypes 763:772 cell line phenotypes 753:772 cell line phenotypes 753:772 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 0 30 theme Caudal 119:124 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 0 31 theme Cell 47:50 arg1 Lines 52:56 Colorectal Cancer Cell Lines 29:56 Colorectal Cancer Cell Lines 29:56 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 7 32 theme glycobiological 1363:1377 arg1 system 1392:1397 a glycobiological cancer model system 1361:1397 a glycobiological cancer model system 1361:1397 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 7 32 theme glycobiological 1363:1377 arg1 use 1354:1356 their use 1348:1356 their use 1348:1356 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 3 33 theme CRC 405:407 arg1 lines 414:418 CRC cell lines 405:418 CRC cell lines with regard to their glycosylation 405:453 However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. 26537799 5 34 theme optimized 789:797 arg1 release 848:854 an optimized, high-throughput membrane-based enzymatic glycan release 786:854 release 848:854 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 4 35 theme N-glycan 536:543 arg1 profiles 545:552 the N-glycan profiles 532:552 the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites 532:633 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 5 36 theme high-throughput 800:814 arg1 release 848:854 an optimized, high-throughput membrane-based enzymatic glycan release 786:854 release 848:854 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 7 37 dep large 1295:1299 arg1 overlap 1301:1307 overlap 1301:1307 overlap 1301:1307 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 7 38 theme N-glycan 1213:1220 arg1 profiles 1222:1229 tissue-derived N-glycan profiles 1198:1229 tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content 1198:1280 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 0 39 theme Cancer 40:45 arg1 Lines 52:56 Colorectal Cancer Cell Lines 29:56 Colorectal Cancer Cell Lines 29:56 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 9 40 theme mRNA 1627:1630 arg1 levels 1643:1648 mRNA expression levels 1627:1648 mRNA expression levels of glycosyltransferases 1627:1672 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 10 41 dep correlate 1963:1971 arg1 both 1958:1961 both 1958:1961 both 1958:1961 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 8 42 theme high-mannose 1436:1447 arg1 N-glycans 1449:1457 the high-mannose N-glycans 1432:1457 the high-mannose N-glycans 1432:1457 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 42 theme high-mannose 1436:1447 arg1 present 1520:1526 present 1520:1526 present 1520:1526 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 0 43 theme Lines 52:56 arg1 Profiling 16:24 N-glycosylation Profiling 0:24 N-glycosylation Profiling of Colorectal Cancer Cell Lines 0:56 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 6 44 theme lines 1154:1158 arg1 patterns 1133:1140 the N-glycosylation patterns 1113:1140 the N-glycosylation patterns of CRC cell lines 1113:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 10 45 gly fucosylated 1894:1904 arg1 glycans 1906:1912 highly fucosylated glycans and CDX1 and/or villin mRNA expression 1887:1951 highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation 1887:1997 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 10 46 theme line 1787:1790 arg1 features 1806:1813 CRC cell line glycosylation features 1778:1813 CRC cell line glycosylation features 1778:1813 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 9 47 theme glycosyltransferases 1653:1672 arg1 levels 1643:1648 mRNA expression levels 1627:1648 mRNA expression levels of glycosyltransferases 1627:1672 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 6 48 theme CRC 1145:1147 arg1 lines 1154:1158 CRC cell lines 1145:1158 CRC cell lines 1145:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 10 49 theme mRNA 1937:1940 arg1 expression 1942:1951 villin mRNA expression 1930:1951 villin mRNA expression 1930:1951 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 10 50 theme cell 1782:1785 arg1 features 1806:1813 CRC cell line glycosylation features 1778:1813 CRC cell line glycosylation features 1778:1813 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 5 51 theme small 860:864 arg1 amounts 873:879 small sample amounts 860:879 small sample amounts 860:879 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 10 52 theme cell 1978:1981 arg1 differentiation 1983:1997 cell differentiation 1978:1997 cell differentiation 1978:1997 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 4 53 theme different 560:568 arg1 lines 579:583 25 different CRC cell lines 557:583 25 different CRC cell lines 557:583 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 2 54 theme cell 282:285 arg1 lines 287:291 CRC cell lines 278:291 CRC cell lines 278:291 CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. 26537799 8 55 attach present 1520:1526 arg2 present 1520:1526 present 1520:1526 present 1520:1526 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 55 attach present 1520:1526 arg1 surface 1540:1546 the cell surface 1531:1546 the cell surface 1531:1546 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 55 attach present 1520:1526 arg2 N-glycans 1449:1457 the high-mannose N-glycans 1432:1457 the high-mannose N-glycans 1432:1457 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 4 56 theme cell 574:577 arg1 lines 579:583 25 different CRC cell lines 557:583 25 different CRC cell lines 557:583 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 0 57 with Association 66:76 arg1 Differentiation 99:113 Differentiation 99:113 Differentiation 99:113 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 0 57 with Association 66:76 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 10 58 theme glycosylation 1792:1804 arg1 features 1806:1813 CRC cell line glycosylation features 1778:1813 CRC cell line glycosylation features 1778:1813 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 7 59 theme cell 1165:1168 arg1 profiles 1175:1182 CRC cell line profiles 1161:1182 CRC cell line profiles 1161:1182 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 6 60 theme Released 882:889 arg1 glycans 891:897 Released glycans 882:897 Released glycans 882:897 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 9 61 theme structural 1722:1731 arg1 analysis 1733:1740 the structural analysis 1718:1740 the structural analysis of glycans 1718:1751 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 7 62 theme high-mannose 1252:1263 arg1 content 1274:1280 their high-mannose N-glycan content 1246:1280 their high-mannose N-glycan content 1246:1280 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 11 63 theme in-depth 2110:2117 arg1 experiments 2119:2129 more in-depth experiments 2105:2129 more in-depth experiments on glycan function and regulation 2105:2163 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 5 64 theme sample 866:871 arg1 amounts 873:879 small sample amounts 860:879 small sample amounts 860:879 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 10 65 theme cell 1827:1830 arg1 markers 1832:1838 cancer cell markers 1820:1838 cancer cell markers 1820:1838 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 5 66 theme membrane-based 816:829 arg1 release 848:854 an optimized, high-throughput membrane-based enzymatic glycan release 786:854 release 848:854 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 6 67 theme N-acetylneuraminic 977:994 arg1 acids 996:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 7 68 theme type 1321:1324 arg1 N-glycans 1326:1334 complex type N-glycans 1313:1334 complex type N-glycans 1313:1334 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 0 69 theme CDX1 142:145 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 11 70 theme new 2031:2033 arg1 insights 2035:2042 new insights 2031:2042 new insights into CRC-associated glycan changes 2031:2077 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 5 71 theme glycan 841:846 arg1 release 848:854 an optimized, high-throughput membrane-based enzymatic glycan release 786:854 release 848:854 We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. 26537799 11 72 theme glycan 2134:2139 arg1 function 2141:2148 glycan function 2134:2148 glycan function 2134:2148 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 4 73 theme metastatic 618:627 arg1 sites 629:633 metastatic sites 618:633 metastatic sites 618:633 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 0 74 theme /Villin 147:153 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 10 75 theme CRC 1778:1780 arg1 features 1806:1813 CRC cell line glycosylation features 1778:1813 CRC cell line glycosylation features 1778:1813 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 4 76 theme glycobiological 679:693 arg1 systems 707:713 glycobiological tumor model systems 679:713 glycobiological tumor model systems 679:713 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 11 77 theme glycan 2064:2069 arg1 changes 2071:2077 CRC-associated glycan changes 2049:2077 CRC-associated glycan changes 2049:2077 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 4 78 theme model 701:705 arg1 systems 707:713 glycobiological tumor model systems 679:713 glycobiological tumor model systems 679:713 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 7 79 theme N-glycan 1265:1272 arg1 content 1274:1280 their high-mannose N-glycan content 1246:1280 their high-mannose N-glycan content 1246:1280 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 7 80 link tissue-derived 1198:1211 arg1 profiles 1222:1229 tissue-derived N-glycan profiles 1198:1229 tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content 1198:1280 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 6 81 theme MALDI-TOF 1043:1051 arg1 -MS. 1058:1061 MALDI-TOF(/TOF)-MS. 1043:1061 MALDI-TOF(/TOF)-MS. 1043:1061 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 7 82 theme model 1386:1390 arg1 system 1392:1397 a glycobiological cancer model system 1361:1397 a glycobiological cancer model system 1361:1397 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 7 82 theme model 1386:1390 arg1 use 1354:1356 their use 1348:1356 their use 1348:1356 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 9 83 theme obtained 1553:1560 arg1 features 1585:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 9 84 theme cell 1566:1569 arg1 features 1585:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 8 85 located present 1520:1526 arg2 present 1520:1526 present 1520:1526 present 1520:1526 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 85 located present 1520:1526 arg1 surface 1540:1546 the cell surface 1531:1546 the cell surface 1531:1546 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 85 located present 1520:1526 arg2 N-glycans 1449:1457 the high-mannose N-glycans 1432:1457 the high-mannose N-glycans 1432:1457 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 6 86 gly N-glycosylation 1117:1131 arg1 lines 1154:1158 CRC cell lines 1145:1158 CRC cell lines 1145:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 0 87 theme Type 126:129 arg1 Expression 160:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression 119:169 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 1 88 theme colorectal 196:205 arg1 CRC 215:217 CRC 215:217 CRC 215:217 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 1 88 theme colorectal 196:205 arg1 cancer 207:212 colorectal cancer 196:212 colorectal cancer (CRC) 196:218 Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. 26537799 9 89 theme N-glycan 1576:1583 arg1 features 1585:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features 1549:1592 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 4 90 theme cell 753:756 arg1 phenotypes 763:772 cell line phenotypes 753:772 cell line phenotypes 753:772 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 7 91 theme tissue-derived 1198:1211 arg1 profiles 1222:1229 tissue-derived N-glycan profiles 1198:1229 tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content 1198:1280 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 6 92 theme pronounced 1082:1091 arg1 differences 1093:1103 pronounced differences 1082:1103 pronounced differences between the N-glycosylation patterns of CRC cell lines 1082:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 3 93 theme cell 409:412 arg1 lines 414:418 CRC cell lines 405:418 CRC cell lines with regard to their glycosylation 405:453 However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. 26537799 0 94 theme Colorectal 29:38 arg1 Lines 52:56 Colorectal Cancer Cell Lines 29:56 Colorectal Cancer Cell Lines 29:56 N-glycosylation Profiling of Colorectal Cancer Cell Lines Reveals Association of Fucosylation with Differentiation and Caudal Type Homebox 1 (CDX1)/Villin mRNA Expression. 26537799 10 95 theme villin 1930:1935 arg1 expression 1942:1951 villin mRNA expression 1930:1951 villin mRNA expression 1930:1951 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 7 96 theme line 1170:1173 arg1 profiles 1175:1182 CRC cell line profiles 1161:1182 CRC cell line profiles 1161:1182 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 11 97 from experiments 2119:2129 arg1 function 2141:2148 glycan function 2134:2148 glycan function 2134:2148 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 11 97 from experiments 2119:2129 arg1 regulation 2154:2163 regulation 2154:2163 regulation 2154:2163 Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation. 26537799 7 98 theme CRC 1161:1163 arg1 profiles 1175:1182 CRC cell line profiles 1161:1182 CRC cell line profiles 1161:1182 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 9 99 theme expression 1632:1641 arg1 levels 1643:1648 mRNA expression levels 1627:1648 mRNA expression levels of glycosyltransferases 1627:1672 The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. 26537799 7 100 theme complex 1313:1319 arg1 N-glycans 1326:1334 complex type N-glycans 1313:1334 complex type N-glycans 1313:1334 CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. 26537799 10 101 with correlation 1763:1773 arg1 phenotypes 1844:1853 phenotypes 1844:1853 phenotypes 1844:1853 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 10 101 with correlation 1763:1773 arg1 markers 1832:1838 cancer cell markers 1820:1838 cancer cell markers 1820:1838 Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. 26537799 6 102 link α2,6-linked 965:975 arg1 acids 996:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 α2,3- and α2,6-linked N-acetylneuraminic acids 955:1000 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 6 103 theme cell 1149:1152 arg1 lines 1154:1158 CRC cell lines 1145:1158 CRC cell lines 1145:1158 Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. 26537799 2 104 theme biological 335:344 arg1 changes 346:352 these (glyco)biological changes 322:352 these (glyco)biological changes 322:352 CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. 26537799 2 105 used used 308:311 arg2 lines 287:291 CRC cell lines 278:291 CRC cell lines 278:291 CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. 26537799 4 106 theme primary 599:605 arg1 tumors 607:612 primary tumors 599:612 primary tumors 599:612 Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. 26537799 8 107 from surface 1540:1546 arg1 N-glycans 1449:1457 the high-mannose N-glycans 1432:1457 the high-mannose N-glycans 1432:1457 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26537799 8 107 from surface 1540:1546 arg1 present 1520:1526 present 1520:1526 present 1520:1526 Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. 26240146 8 0 theme Site-specific 1229:1241 arg1 analysis 1259:1266 Site-specific N-glycosylation analysis 1229:1266 Site-specific N-glycosylation analysis 1229:1266 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 4 1 theme exchange 547:554 arg1 approach 563:570 a hydrogen/deuterium exchange and MS approach 526:570 a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms 526:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 3 2 gly heterogeneity 455:467 arg1 carbohydrates 495:507 carbohydrates 495:507 carbohydrates 495:507 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 7 3 theme Hydrogen/deuterium 1117:1134 arg1 exchange-MS 1136:1146 Hydrogen/deuterium exchange-MS 1117:1146 Hydrogen/deuterium exchange-MS 1117:1146 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 8 4 theme N-glycosylation 1243:1257 arg1 analysis 1259:1266 Site-specific N-glycosylation analysis 1229:1266 Site-specific N-glycosylation analysis 1229:1266 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 6 5 dep alteration 1072:1081 arg1 response 1056:1063 response 1056:1063 response 1056:1063 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 10 6 theme glycoprotein 1575:1586 arg1 diagnostics 1611:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 0 7 from Recognition 0:10 arg1 gonadotropin 47:58 human chorionic gonadotropin 31:58 human chorionic gonadotropin 31:58 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 7 8 theme subunit 1193:1199 arg1 epitope 1208:1214 the epitope 1204:1214 the epitope for MCA1024 1204:1226 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 7 8 theme subunit 1193:1199 arg1 β65-83 1173:1178 the peptide β65-83 1161:1178 the peptide β65-83 of the hCG β subunit 1161:1199 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 5 9 theme Hyperglycosylated 680:696 arg1 hCG 698:700 Hyperglycosylated hCG 680:700 Hyperglycosylated hCG 680:700 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 10 10 gly glycoprotein 1575:1586 arg1 glycoprotein 1575:1586 glycoprotein biomarker-based cancer diagnostics 1575:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 0 11 theme interaction 95:105 arg1 motifs 107:112 their interaction motifs 89:112 their interaction motifs 89:112 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 4 12 gly glycoforms 668:677 arg1 hCG 664:666 different hCG glycoforms 654:677 different hCG glycoforms 654:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 6 13 link N-linked 1090:1097 arg1 oligosaccharides 1099:1114 hCG N-linked oligosaccharides 1086:1114 hCG N-linked oligosaccharides 1086:1114 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 4 14 theme antibodies 635:644 arg1 binding 624:630 the binding 620:630 the binding of antibodies against different hCG glycoforms 620:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 5 15 theme oligosaccharides 791:806 arg1 branched 833:840 branched 833:840 branched 833:840 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 15 theme oligosaccharides 791:806 arg1 structure 765:773 the structure 761:773 the structure of its N-linked oligosaccharides 761:806 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 16 theme antibodies 886:895 arg1 kinetics 861:868 The binding kinetics 849:868 The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG 849:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 17 gly Hyperglycosylated 680:696 arg1 hCG 698:700 Hyperglycosylated hCG 680:700 Hyperglycosylated hCG 680:700 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 10 18 theme combined 1637:1644 arg1 immunoassays 1646:1657 combined immunoassays 1637:1657 combined immunoassays that can determine the level of protein 1637:1697 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 7 19 theme hCG 1187:1189 arg1 subunit 1193:1199 the hCG β subunit 1183:1199 the hCG β subunit 1183:1199 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 10 20 theme protein 1691:1697 arg1 level 1682:1686 the level 1678:1686 the level of protein 1678:1697 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 4 21 theme hydrogen/deuterium 528:545 arg1 exchange 547:554 hydrogen/deuterium exchange 528:554 hydrogen/deuterium exchange 528:554 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 4 22 from effect 591:596 arg1 binding 624:630 the binding 620:630 the binding of antibodies against different hCG glycoforms 620:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 8 23 from Asn-30 1322:1327 arg1 oligosaccharides 1291:1306 N-linked oligosaccharides 1282:1306 N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit 1282:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 2 24 theme diagnostic 290:299 arg1 accuracy 301:308 diagnostic accuracy 290:308 diagnostic accuracy 290:308 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 8 25 theme β 1336:1336 arg1 subunit 1338:1344 the β subunit 1332:1344 the β subunit 1332:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 5 26 theme binding 853:859 arg1 kinetics 861:868 The binding kinetics 849:868 The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG 849:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 9 27 theme structural 1448:1457 arg1 change 1459:1464 the structural change 1444:1464 the structural change of N-linked oligosaccharides 1444:1493 These results prove that some antibodies are sensitive to the structural change of N-linked oligosaccharides, whereas others are not affected by N-glycosylation. 26240146 10 28 theme N-glycosylation 1725:1739 arg1 degree 1715:1720 the degree 1711:1720 the degree of N-glycosylation 1711:1739 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 5 29 gly hyperglycosylated 932:948 arg1 hCG 950:952 hyperglycosylated hCG 932:952 hyperglycosylated hCG 932:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 1 30 theme important 180:188 arg1 role 190:193 an important role 177:193 an important role 177:193 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 8 31 theme N-linked 1282:1289 arg1 oligosaccharides 1291:1306 N-linked oligosaccharides 1282:1306 N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit 1282:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 4 32 theme N-glycosylation 601:615 arg1 effect 591:596 the effect 587:596 the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms 587:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 3 33 dep heterogeneity 455:467 arg1 the 451:453 the 451:453 the 451:453 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 9 34 theme N-linked 1469:1476 arg1 oligosaccharides 1478:1493 N-linked oligosaccharides 1469:1493 N-linked oligosaccharides 1469:1493 These results prove that some antibodies are sensitive to the structural change of N-linked oligosaccharides, whereas others are not affected by N-glycosylation. 26240146 2 35 theme cancers 337:343 arg1 sensitivity 314:324 sensitivity 314:324 sensitivity 314:324 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 2 35 theme cancers 337:343 arg1 accuracy 301:308 diagnostic accuracy 290:308 diagnostic accuracy 290:308 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 8 36 theme binding 1357:1363 arg1 affinity 1365:1372 the binding affinity 1353:1372 the binding affinity of MCA1024 1353:1383 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 3 37 theme low 473:475 arg1 immunogenicity 477:490 low immunogenicity 473:490 low immunogenicity 473:490 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 3 38 theme N-linked 402:409 arg1 oligosaccharides 411:426 the N-linked oligosaccharides 398:426 the N-linked oligosaccharides 398:426 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 7 39 theme peptide 1165:1171 arg1 epitope 1208:1214 the epitope 1204:1214 the epitope for MCA1024 1204:1226 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 7 39 theme peptide 1165:1171 arg1 β65-83 1173:1178 the peptide β65-83 1161:1178 the peptide β65-83 of the hCG β subunit 1161:1199 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 8 40 link N-linked 1282:1289 arg1 oligosaccharides 1291:1306 N-linked oligosaccharides 1282:1306 N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit 1282:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 2 41 theme related 329:335 arg1 cancers 337:343 related cancers 329:343 related cancers 329:343 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 7 42 theme β 1191:1191 arg1 subunit 1193:1199 the hCG β subunit 1183:1199 the hCG β subunit 1183:1199 Hydrogen/deuterium exchange-MS reveals that the peptide β65-83 of the hCG β subunit is the epitope for MCA1024. 26240146 5 43 theme anti-hCG 877:884 arg1 MCA329 897:902 MCA329 897:902 MCA329 897:902 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 43 theme anti-hCG 877:884 arg1 antibodies 886:895 the anti-hCG antibodies MCA329 and MCA1024 873:914 the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG 873:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 43 theme anti-hCG 877:884 arg1 MCA1024 908:914 MCA1024 908:914 MCA1024 908:914 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 0 44 theme N-glycoforms 15:26 arg1 Recognition 0:10 Recognition 0:10 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 0:113 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 1 45 gly glycosylation 119:131 arg1 hCG 166:168 hCG 166:168 hCG 166:168 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 1 45 gly glycosylation 119:131 arg1 gonadotropin 152:163 human chorionic gonadotropin 136:163 human chorionic gonadotropin (hCG) 136:169 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 1 46 theme reproductive 198:209 arg1 tumors 211:216 reproductive tumors 198:216 reproductive tumors 198:216 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 0 47 theme chorionic 37:45 arg1 gonadotropin 47:58 human chorionic gonadotropin 31:58 human chorionic gonadotropin 31:58 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 6 48 theme MCA1024 1023:1029 arg1 affinity 1011:1018 The binding affinity 999:1018 The binding affinity of MCA1024 999:1029 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 8 49 theme MCA1024 1377:1383 arg1 affinity 1365:1372 the binding affinity 1353:1372 the binding affinity of MCA1024 1353:1383 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 3 50 theme carbohydrates 495:507 arg1 immunogenicity 477:490 low immunogenicity 473:490 low immunogenicity 473:490 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 3 50 theme carbohydrates 495:507 arg1 heterogeneity 455:467 heterogeneity 455:467 heterogeneity 455:467 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 0 51 theme human 31:35 arg1 gonadotropin 47:58 human chorionic gonadotropin 31:58 human chorionic gonadotropin 31:58 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 10 52 theme cancer 1604:1609 arg1 diagnostics 1611:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 2 53 theme N-glycosylation 233:247 arg1 alteration 249:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 9 54 link N-linked 1469:1476 arg1 oligosaccharides 1478:1493 N-linked oligosaccharides 1469:1493 N-linked oligosaccharides 1469:1493 These results prove that some antibodies are sensitive to the structural change of N-linked oligosaccharides, whereas others are not affected by N-glycosylation. 26240146 5 55 theme hyperglycosylated 932:948 arg1 hCG 950:952 hyperglycosylated hCG 932:952 hyperglycosylated hCG 932:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 56 theme invasive 733:740 arg1 mole 742:745 invasive mole 733:745 invasive mole patients 733:754 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 2 57 theme hCG 229:231 arg1 alteration 249:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 4 58 gly N-glycosylation 601:615 arg1 antibodies 635:644 antibodies 635:644 antibodies against different hCG glycoforms 635:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 4 59 theme MS 560:561 arg1 approach 563:570 a hydrogen/deuterium exchange and MS approach 526:570 a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms 526:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 6 60 theme binding 1003:1009 arg1 affinity 1011:1018 The binding affinity 999:1018 The binding affinity of MCA1024 999:1029 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 2 61 dep accuracy 301:308 arg1 the 286:288 the 286:288 the 286:288 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 8 62 from oligosaccharides 1291:1306 arg1 subunit 1338:1344 the β subunit 1332:1344 the β subunit 1332:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 2 63 theme Detecting 219:227 arg1 alteration 249:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration 219:258 Detecting hCG N-glycosylation alteration may significantly improve the diagnostic accuracy and sensitivity of related cancers. 26240146 5 64 theme biolayer 974:981 arg1 interferometry 983:996 biolayer interferometry 974:996 biolayer interferometry 974:996 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 9 65 dep sensitive 1431:1439 arg1 whereas 1496:1502 whereas 1496:1502 whereas 1496:1502 These results prove that some antibodies are sensitive to the structural change of N-linked oligosaccharides, whereas others are not affected by N-glycosylation. 26240146 9 66 theme oligosaccharides 1478:1493 arg1 change 1459:1464 the structural change 1444:1464 the structural change of N-linked oligosaccharides 1444:1493 These results prove that some antibodies are sensitive to the structural change of N-linked oligosaccharides, whereas others are not affected by N-glycosylation. 26240146 5 67 theme patients 747:754 arg1 urine 724:728 the urine 720:728 the urine of invasive mole patients 720:754 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 3 68 link N-linked 402:409 arg1 oligosaccharides 411:426 the N-linked oligosaccharides 398:426 the N-linked oligosaccharides 398:426 However, developing an immunoassay directly against the N-linked oligosaccharides is unlikely because of the heterogeneity and low immunogenicity of carbohydrates. 26240146 6 69 theme N-linked 1090:1097 arg1 oligosaccharides 1099:1114 hCG N-linked oligosaccharides 1086:1114 hCG N-linked oligosaccharides 1086:1114 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 1 70 theme human 136:140 arg1 hCG 166:168 hCG 166:168 hCG 166:168 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 1 70 theme human 136:140 arg1 gonadotropin 152:163 human chorionic gonadotropin 136:163 human chorionic gonadotropin (hCG) 136:169 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 4 71 theme hCG 664:666 arg1 glycoforms 668:677 different hCG glycoforms 654:677 different hCG glycoforms 654:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 5 72 link N-linked 782:789 arg1 oligosaccharides 791:806 its N-linked oligosaccharides 778:806 its N-linked oligosaccharides 778:806 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 10 73 theme biomarker-based 1588:1602 arg1 diagnostics 1611:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 glycoprotein biomarker-based cancer diagnostics 1575:1621 It is promising to improve glycoprotein biomarker-based cancer diagnostics by developing combined immunoassays that can determine the level of protein and measure the degree of N-glycosylation simultaneously. 26240146 1 74 theme chorionic 142:150 arg1 hCG 166:168 hCG 166:168 hCG 166:168 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 1 74 theme chorionic 142:150 arg1 gonadotropin 152:163 human chorionic gonadotropin 136:163 human chorionic gonadotropin (hCG) 136:169 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 4 75 theme different 654:662 arg1 glycoforms 668:677 different hCG glycoforms 654:677 different hCG glycoforms 654:677 Here, we report a hydrogen/deuterium exchange and MS approach to investigate the effect of N-glycosylation on the binding of antibodies against different hCG glycoforms. 26240146 1 76 theme gonadotropin 152:163 arg1 glycosylation 119:131 The glycosylation 115:131 The glycosylation of human chorionic gonadotropin (hCG) 115:169 The glycosylation of human chorionic gonadotropin (hCG) plays an important role in reproductive tumors. 26240146 6 77 theme hCG 1086:1088 arg1 oligosaccharides 1099:1114 hCG N-linked oligosaccharides 1086:1114 hCG N-linked oligosaccharides 1086:1114 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 0 78 theme monoclonal 63:72 arg1 antibodies 74:83 monoclonal antibodies 63:83 monoclonal antibodies 63:83 Recognition of N-glycoforms in human chorionic gonadotropin by monoclonal antibodies and their interaction motifs. 26240146 8 79 from Asn-13 1311:1316 arg1 oligosaccharides 1291:1306 N-linked oligosaccharides 1282:1306 N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit 1282:1344 Site-specific N-glycosylation analysis suggests that N-linked oligosaccharides at Asn-13 and Asn-30 on the β subunit affect the binding affinity of MCA1024. 26240146 5 80 dep antibodies 886:895 arg1 MCA329 897:902 MCA329 897:902 MCA329 897:902 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 80 dep antibodies 886:895 arg1 antibodies 886:895 the anti-hCG antibodies MCA329 and MCA1024 873:914 the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG 873:952 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 80 dep antibodies 886:895 arg1 MCA1024 908:914 MCA1024 908:914 MCA1024 908:914 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 6 81 theme oligosaccharides 1099:1114 arg1 alteration 1072:1081 the alteration 1068:1081 the alteration of hCG N-linked oligosaccharides 1068:1114 The binding affinity of MCA1024 changed significantly in response to the alteration of hCG N-linked oligosaccharides. 26240146 5 82 theme mole 742:745 arg1 patients 747:754 invasive mole patients 733:754 invasive mole patients 733:754 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 26240146 5 83 theme N-linked 782:789 arg1 oligosaccharides 791:806 its N-linked oligosaccharides 778:806 its N-linked oligosaccharides 778:806 Hyperglycosylated hCG was purified from the urine of invasive mole patients, and the structure of its N-linked oligosaccharides was confirmed to be more branched by MS. The binding kinetics of the anti-hCG antibodies MCA329 and MCA1024 against hCG and hyperglycosylated hCG were compared using biolayer interferometry. 27932460 1 0 theme pathway 181:187 arg1 Disruption 147:156 Disruption 147:156 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan 147:242 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 2 1 link O-linked 299:306 arg1 POMGNT2 357:363 POMGNT2 357:363 POMGNT2 357:363 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 2 1 link O-linked 299:306 arg1 β-1,4-N-acetylglucosaminyltransferase 316:352 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 291:354 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) 291:364 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 0 2 theme Gatekeeper 81:90 arg1 Enzyme 92:97 a Gatekeeper Enzyme 79:97 a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan 79:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 2 theme Gatekeeper 81:90 arg1 β-1,4-N-Acetylglucosaminyl-transferase 25:62 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 0:64 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) 0:74 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 6 3 theme conserved 1238:1246 arg1 acids 1254:1258 the conserved amino acids 1234:1258 the conserved amino acids 1234:1258 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 5 4 theme POMGNT1-only 1043:1054 arg1 site 1056:1059 a POMGNT1-only site 1041:1059 a POMGNT1-only site 1041:1059 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 5 5 gly O-mannosylated 1090:1103 arg1 peptide 1105:1111 the O-mannosylated peptide 1086:1111 the O-mannosylated peptide 1086:1111 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 0 6 theme Functional 103:112 arg1 Glycosylation 114:126 Functional Glycosylation 103:126 Functional Glycosylation of α-Dystroglycan 103:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 7 7 theme functional 1505:1514 arg1 structures 1494:1503 glycan structures 1487:1503 glycan structures functional for binding laminin globular domain-containing proteins 1487:1570 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 3 8 link O-linked 564:571 arg1 β-1,2-N-acetylglucosaminyltransferase 581:617 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 556:619 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 556:629 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 8 link O-linked 564:571 arg1 POMGNT1 622:628 POMGNT1 622:628 POMGNT1 622:628 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 9 theme other 663:667 arg1 structures 684:693 other various glycan structures 663:693 other various glycan structures present on α-dystroglycan of unknown function 663:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 7 10 from majority 1414:1421 arg1 proteins 1450:1457 proteins 1450:1457 proteins 1450:1457 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 1 11 gly glycosylation 212:224 arg1 α-dystroglycan 229:242 α-dystroglycan 229:242 α-dystroglycan 229:242 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 3 12 theme various 669:675 arg1 structures 684:693 other various glycan structures 663:693 other various glycan structures present on α-dystroglycan of unknown function 663:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 4 13 theme amino 865:869 arg1 selectivity 876:886 significant primary amino acid selectivity 845:886 significant primary amino acid selectivity 845:886 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 7 14 theme POMGNT2 1324:1330 arg1 selectivity 1309:1319 the selectivity 1305:1319 the selectivity of POMGNT2 1305:1330 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 4 15 theme primary 857:863 arg1 selectivity 876:886 significant primary amino acid selectivity 845:886 significant primary amino acid selectivity 845:886 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 5 16 theme vertebrate 977:986 arg1 species 988:994 59 vertebrate species 974:994 59 vertebrate species 974:994 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 3 17 attach present 695:701 arg2 structures 684:693 other various glycan structures 663:693 other various glycan structures present on α-dystroglycan of unknown function 663:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 17 attach present 695:701 arg1 α-dystroglycan 706:719 α-dystroglycan 706:719 α-dystroglycan of unknown function 706:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 6 18 theme efficient 1190:1198 arg1 substrate 1200:1208 a less efficient substrate 1183:1208 a less efficient substrate for POMGNT2 1183:1220 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 6 18 theme efficient 1190:1198 arg1 glycopeptide 1167:1178 an acceptor glycopeptide 1155:1178 an acceptor glycopeptide 1155:1178 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 3 19 theme O-linked 564:571 arg1 β-1,2-N-acetylglucosaminyltransferase 581:617 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 556:619 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 556:629 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 19 theme O-linked 564:571 arg1 POMGNT1 622:628 POMGNT1 622:628 POMGNT1 622:628 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 5 20 from motif 951:955 arg1 α-dystroglycan 1000:1013 α-dystroglycan 1000:1013 α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2 1000:1138 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 1 21 theme functional 201:210 arg1 glycosylation 212:224 functional glycosylation 201:224 functional glycosylation of α-dystroglycan 201:242 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 0 22 theme O-Linked 8:15 arg1 β-1,4-N-Acetylglucosaminyl-transferase 25:62 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 0:64 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) 0:74 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 22 theme O-Linked 8:15 arg1 POMGNT2 67:73 POMGNT2 67:73 POMGNT2 67:73 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 22 theme O-Linked 8:15 arg1 Enzyme 92:97 a Gatekeeper Enzyme 79:97 a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan 79:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 2 23 from structure 425:433 arg1 α-dystroglycan 438:451 α-dystroglycan 438:451 α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses 438:538 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 3 24 theme present 695:701 arg1 structures 684:693 other various glycan structures 663:693 other various glycan structures present on α-dystroglycan of unknown function 663:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 0 25 theme Protein 0:6 arg1 β-1,4-N-Acetylglucosaminyl-transferase 25:62 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 0:64 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) 0:74 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 25 theme Protein 0:6 arg1 POMGNT2 67:73 POMGNT2 67:73 POMGNT2 67:73 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 25 theme Protein 0:6 arg1 Enzyme 92:97 a Gatekeeper Enzyme 79:97 a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan 79:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 7 26 gly O-mannosylated 1426:1439 arg1 sites 1441:1445 O-mannosylated sites 1426:1445 O-mannosylated sites 1426:1445 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 2 27 theme first 380:384 arg1 step 386:389 the first step 376:389 the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses 376:538 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 4 28 theme O-mannosylation 905:919 arg1 O-mannosylation 905:919 O-mannosylation 905:919 O-mannosylation 905:919 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 4 28 theme O-mannosylation 905:919 arg1 site 897:900 the site 893:900 the site of O-mannosylation 893:919 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 4 29 theme O-mannosylated 795:808 arg1 peptides 810:817 O-mannosylated peptides 795:817 O-mannosylated peptides 795:817 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 7 30 theme glycan 1487:1492 arg1 structures 1494:1503 glycan structures 1487:1503 glycan structures functional for binding laminin globular domain-containing proteins 1487:1570 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 0 31 gly Glycosylation 114:126 arg1 α-Dystroglycan 131:144 α-Dystroglycan 131:144 α-Dystroglycan 131:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 7 32 theme laminin 1528:1534 arg1 proteins 1563:1570 laminin globular domain-containing proteins 1528:1570 laminin globular domain-containing proteins 1528:1570 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 1 33 theme α-dystroglycan 229:242 arg1 glycosylation 212:224 functional glycosylation 201:224 functional glycosylation of α-dystroglycan 201:242 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 0 34 theme Mannose 17:23 arg1 β-1,4-N-Acetylglucosaminyl-transferase 25:62 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 0:64 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) 0:74 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 34 theme Mannose 17:23 arg1 POMGNT2 67:73 POMGNT2 67:73 POMGNT2 67:73 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 34 theme Mannose 17:23 arg1 Enzyme 92:97 a Gatekeeper Enzyme 79:97 a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan 79:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 0 35 theme α-Dystroglycan 131:144 arg1 Glycosylation 114:126 Functional Glycosylation 103:126 Functional Glycosylation of α-Dystroglycan 103:144 Protein O-Linked Mannose β-1,4-N-Acetylglucosaminyl-transferase 2 (POMGNT2) Is a Gatekeeper Enzyme for Functional Glycosylation of α-Dystroglycan. 27932460 5 36 theme O-mannosylated 1090:1103 arg1 peptide 1105:1111 the O-mannosylated peptide 1086:1111 the O-mannosylated peptide 1086:1111 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 7 37 theme gatekeeper 1376:1385 arg1 enzyme 1387:1392 a gatekeeper enzyme 1374:1392 a gatekeeper enzyme 1374:1392 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 7 37 theme gatekeeper 1376:1385 arg1 enzyme 1354:1359 this enzyme 1349:1359 this enzyme 1349:1359 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 3 38 theme mannose 573:579 arg1 β-1,2-N-acetylglucosaminyltransferase 581:617 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 556:619 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 556:629 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 38 theme mannose 573:579 arg1 POMGNT1 622:628 POMGNT1 622:628 POMGNT1 622:628 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 7 39 theme O-mannosylated 1426:1439 arg1 sites 1441:1445 O-mannosylated sites 1426:1445 O-mannosylated sites 1426:1445 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 4 40 theme significant 845:855 arg1 selectivity 876:886 significant primary amino acid selectivity 845:886 significant primary amino acid selectivity 845:886 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 3 41 theme unknown 724:730 arg1 function 732:739 unknown function 724:739 unknown function 724:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 2 42 theme matrix 499:504 arg1 proteins 506:513 extracellular matrix proteins 485:513 extracellular matrix proteins 485:513 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 4 43 gly O-mannosylated 795:808 arg1 peptides 810:817 O-mannosylated peptides 795:817 O-mannosylated peptides 795:817 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 7 44 theme domain-containing 1545:1561 arg1 proteins 1563:1570 laminin globular domain-containing proteins 1528:1570 laminin globular domain-containing proteins 1528:1570 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 3 45 theme function 732:739 arg1 α-dystroglycan 706:719 α-dystroglycan 706:719 α-dystroglycan of unknown function 706:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 2 46 theme extracellular 485:497 arg1 proteins 506:513 extracellular matrix proteins 485:513 extracellular matrix proteins 485:513 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 5 47 theme POMGNT2 934:940 arg1 motif 951:955 a POMGNT2 acceptor motif 932:955 a POMGNT2 acceptor motif 932:955 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 5 48 dep sufficient 1064:1073 arg1 α-dystroglycan 1000:1013 α-dystroglycan 1000:1013 α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2 1000:1138 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 2 49 theme matriglycan 413:423 arg1 structure 425:433 the functional matriglycan structure 398:433 the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses 398:538 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 5 50 theme acceptor 942:949 arg1 motif 951:955 a POMGNT2 acceptor motif 932:955 a POMGNT2 acceptor motif 932:955 We define a POMGNT2 acceptor motif, conserved among 59 vertebrate species, in α-dystroglycan that when engineered into a POMGNT1-only site is sufficient to convert the O-mannosylated peptide to a substrate for POMGNT2. 27932460 7 51 theme vast 1409:1412 arg1 majority 1414:1421 the vast majority 1405:1421 the vast majority of O-mannosylated sites on proteins 1405:1457 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 2 52 theme functional 402:411 arg1 structure 425:433 the functional matriglycan structure 398:433 the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses 398:538 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 4 53 dep promiscuous 779:789 arg1 whereas 820:826 whereas 820:826 whereas 820:826 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 7 54 theme globular 1536:1543 arg1 proteins 1563:1570 laminin globular domain-containing proteins 1528:1570 laminin globular domain-containing proteins 1528:1570 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 4 55 theme acid 871:874 arg1 selectivity 876:886 significant primary amino acid selectivity 845:886 significant primary amino acid selectivity 845:886 Here, we demonstrate that POMGNT1 is promiscuous for O-mannosylated peptides, whereas POMGNT2 displays significant primary amino acid selectivity near the site of O-mannosylation. 27932460 3 56 theme glycan 677:682 arg1 structures 684:693 other various glycan structures 663:693 other various glycan structures present on α-dystroglycan of unknown function 663:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 1 57 theme congenital 258:267 arg1 dystrophies 278:288 congenital muscular dystrophies 258:288 congenital muscular dystrophies 258:288 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 6 58 theme acids 1254:1258 arg1 acids 1254:1258 the conserved amino acids 1234:1258 the conserved amino acids 1234:1258 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 6 58 theme acids 1254:1258 arg1 two 1227:1229 two 1227:1229 two 1227:1229 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 2 59 theme mannose 308:314 arg1 POMGNT2 357:363 POMGNT2 357:363 POMGNT2 357:363 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 2 59 theme mannose 308:314 arg1 β-1,4-N-acetylglucosaminyltransferase 316:352 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 291:354 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) 291:364 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 6 60 gly glycopeptide 1167:1178 arg2 glycopeptide 1167:1178 an acceptor glycopeptide 1155:1178 an acceptor glycopeptide 1155:1178 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 6 60 gly glycopeptide 1167:1178 arg2 substrate 1200:1208 a less efficient substrate 1183:1208 a less efficient substrate for POMGNT2 1183:1220 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 3 61 theme first 645:649 arg1 step 651:654 the first step 641:654 the first step toward other various glycan structures present on α-dystroglycan of unknown function 641:739 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 1 62 theme muscular 269:276 arg1 dystrophies 278:288 congenital muscular dystrophies 258:288 congenital muscular dystrophies 258:288 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 27932460 3 63 theme protein 556:562 arg1 β-1,2-N-acetylglucosaminyltransferase 581:617 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 556:619 protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 556:629 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 3 63 theme protein 556:562 arg1 POMGNT1 622:628 POMGNT1 622:628 POMGNT1 622:628 Alternatively, protein O-linked mannose β-1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) catalyzes the first step toward other various glycan structures present on α-dystroglycan of unknown function. 27932460 2 64 theme O-linked 299:306 arg1 POMGNT2 357:363 POMGNT2 357:363 POMGNT2 357:363 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 2 64 theme O-linked 299:306 arg1 β-1,4-N-acetylglucosaminyltransferase 316:352 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 291:354 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) 291:364 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 2 65 theme certain 519:525 arg1 arenaviruses 527:538 certain arenaviruses 519:538 certain arenaviruses 519:538 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 6 66 theme amino 1248:1252 arg1 acids 1254:1258 the conserved amino acids 1234:1258 the conserved amino acids 1234:1258 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 7 67 theme sites 1441:1445 arg1 majority 1414:1421 the vast majority 1405:1421 the vast majority of O-mannosylated sites on proteins 1405:1457 These findings begin to define the selectivity of POMGNT2 and suggest that this enzyme functions as a gatekeeper enzyme to prevent the vast majority of O-mannosylated sites on proteins from becoming modified with glycan structures functional for binding laminin globular domain-containing proteins. 27932460 6 68 theme acceptor 1158:1165 arg1 substrate 1200:1208 a less efficient substrate 1183:1208 a less efficient substrate for POMGNT2 1183:1220 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 6 68 theme acceptor 1158:1165 arg1 glycopeptide 1167:1178 an acceptor glycopeptide 1155:1178 an acceptor glycopeptide 1155:1178 Additionally, an acceptor glycopeptide is a less efficient substrate for POMGNT2 when two of the conserved amino acids are replaced. 27932460 2 69 theme Protein 291:297 arg1 POMGNT2 357:363 POMGNT2 357:363 POMGNT2 357:363 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 2 69 theme Protein 291:297 arg1 β-1,4-N-acetylglucosaminyltransferase 316:352 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 291:354 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) 291:364 Protein O-linked mannose β-1,4-N-acetylglucosaminyltransferase 2 (POMGNT2) catalyzes the first step toward the functional matriglycan structure on α-dystroglycan that is responsible for binding extracellular matrix proteins and certain arenaviruses. 27932460 1 70 theme O-mannosylation 165:179 arg1 pathway 181:187 the O-mannosylation pathway 161:187 the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan 161:242 Disruption of the O-mannosylation pathway involved in functional glycosylation of α-dystroglycan gives rise to congenital muscular dystrophies. 25927005 6 0 theme glycans 832:838 arg1 effect 822:827 the effect 818:827 the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells 818:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 9 1 theme glycoprotein 1526:1537 arg1 E2 1539:1540 HCV glycoprotein E2 1522:1540 HCV glycoprotein E2 1522:1540 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 7 2 theme N-glycosylation 1040:1054 arg1 sites 1056:1060 the N-glycosylation sites 1036:1060 the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) 1036:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 2 theme N-glycosylation 1040:1054 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 11 3 theme glycosylation 1931:1943 arg1 sites 1945:1949 single glycosylation sites 1924:1949 single glycosylation sites of HCV E2 1924:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 3 theme glycosylation 1931:1943 arg1 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 4 4 theme envelope 529:536 arg1 proteins 538:545 the virus envelope proteins 519:545 the virus envelope proteins in a cell 519:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 10 5 theme unproductive 1761:1772 arg1 dimers 1779:1784 unproductive E1E2 dimers 1761:1784 unproductive E1E2 dimers 1761:1784 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 3 6 theme understood 432:441 arg1 processes 443:451 the least understood processes 422:451 the least understood processes 422:451 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 6 theme understood 432:441 arg1 release 387:393 release 387:393 release 387:393 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 6 theme understood 432:441 arg1 assembly 374:381 virion assembly 367:381 virion assembly 367:381 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 9 7 theme HEK293T 1551:1557 arg1 cells 1559:1563 human HEK293T cells 1545:1563 human HEK293T cells 1545:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 11 8 contain had 1961:1963 arg2 impact 1968:1973 no impact 1965:1973 no impact 1965:1973 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 8 contain had 1961:1963 arg1 elimination 1909:1919 elimination 1909:1919 elimination of single glycosylation sites of HCV E2 1909:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 7 9 theme point 1017:1021 arg1 mutations 1023:1031 point mutations 1017:1031 point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) 1017:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 10 10 from sites 1728:1732 arg1 elimination 1682:1692 elimination 1682:1692 elimination of glycans at the N1, N2, and N10 sites 1682:1732 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 11 11 theme E2 1958:1959 arg1 sites 1945:1949 single glycosylation sites 1924:1949 single glycosylation sites of HCV E2 1924:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 11 theme E2 1958:1959 arg1 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 7 12 theme genotype 1081:1088 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 12 theme genotype 1081:1088 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 6 13 gly glycoprotein 864:875 arg1 glycoprotein 864:875 ofthe E2 glycoprotein 855:875 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 8 14 from decrease 1449:1456 arg1 number 1465:1470 the number 1461:1470 the number of glycosylation sites 1461:1493 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 7 15 gly N-glycosylation 1040:1054 arg2 sites 1056:1060 the N-glycosylation sites 1036:1060 the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) 1036:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 15 gly N-glycosylation 1040:1054 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 15 gly N-glycosylation 1040:1054 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 15 gly N-glycosylation 1040:1054 arg2 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 10 16 theme dimers 1779:1784 arg1 accumulation 1745:1756 the accumulation 1741:1756 the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells 1741:1893 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 0 17 theme Mammalian 97:105 arg1 cells 107:111 insect and Mammalian cells 86:111 insect and Mammalian cells 86:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 10 18 from accumulation 1745:1756 arg1 cells 1889:1893 insect and mammalian cells 1868:1893 insect and mammalian cells 1868:1893 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 9 19 from cells 1559:1563 arg1 level 1500:1504 The level 1496:1504 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells 1496:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 4 20 theme several 574:580 arg1 sites 596:600 glycansat several glycosylation sites 564:600 glycansat several glycosylation sites of these proteins 564:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 20 theme several 574:580 arg1 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 7 21 theme E2 1077:1078 arg1 sites 1056:1060 the N-glycosylation sites 1036:1060 the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) 1036:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 21 theme E2 1077:1078 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 9 22 gly glycosylation 1618:1630 arg2 sites 1632:1636 the N1 and N8 glycosylation sites 1604:1636 sites 1632:1636 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 6 23 theme ofthe 855:859 arg1 glycoprotein 864:875 ofthe E2 glycoprotein 855:875 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 8 24 theme Sf9 1346:1348 arg1 cells 1357:1361 Sf9 insect cells 1346:1361 Sf9 insect cells 1346:1361 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 10 25 theme productive 1804:1813 arg1 suppression 1824:1834 productive assembly suppression 1804:1834 productive assembly suppression 1804:1834 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 9 26 from N1 1608:1609 arg1 glycans 1593:1599 glycans 1593:1599 glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells 1593:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 26 from N1 1608:1609 arg1 presence 1581:1588 the presence 1577:1588 the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells 1577:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 27 theme N8 1615:1616 arg1 sites 1632:1636 the N1 and N8 glycosylation sites 1604:1636 sites 1632:1636 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 0 28 from proteins 74:81 arg1 cells 107:111 insect and Mammalian cells 86:111 insect and Mammalian cells 86:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 8 29 theme single 1228:1233 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 29 theme single 1228:1233 arg1 sites 1249:1253 the single glycosylation sites 1224:1253 the single glycosylation sites of the E2 glycoprotein 1224:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 10 30 theme virus-like 1839:1848 arg1 particles 1850:1858 virus-like particles 1839:1858 virus-like particles 1839:1858 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 7 31 theme mutant 1133:1138 arg1 proteins 1140:1147 the mutant proteins 1129:1147 the mutant proteins 1129:1147 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 1 32 theme C 128:128 arg1 HCV 137:139 HCV 137:139 HCV 137:139 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 32 theme C 128:128 arg1 virus 130:134 hepatitis C virus 118:134 The hepatitis C virus (HCV) envelope proteins E1 and E2 114:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 6 33 theme virus 929:933 arg1 particles 935:943 virus particles 929:943 virus particles in insect and mammalian cells 929:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 8 34 theme electrophoretic 1374:1388 arg1 mobility 1390:1397 the electrophoretic mobility 1370:1397 the electrophoretic mobility of mutant proteins 1370:1416 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 6 35 from particles 935:943 arg1 cells 969:973 insect and mammalian cells 948:973 insect and mammalian cells 948:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 4 36 theme pivotal 627:633 arg1 role 635:638 a pivotal role 625:638 a pivotal role 625:638 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 10 37 theme mammalian 1879:1887 arg1 cells 1889:1893 insect and mammalian cells 1868:1893 insect and mammalian cells 1868:1893 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 0 38 theme virus 59:63 arg1 proteins 74:81 virus envelope proteins 59:81 virus envelope proteins in insect and Mammalian cells 59:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 11 39 theme sites 1945:1949 arg1 elimination 1909:1919 elimination 1909:1919 elimination of single glycosylation sites of HCV E2 1909:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 6 40 theme complexes 915:923 arg1 particles 935:943 virus particles 929:943 virus particles in insect and mammalian cells 929:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 6 40 theme complexes 915:923 arg1 formation 878:886 formation 878:886 formation of functional glycoprotein complexes 878:923 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 6 40 theme complexes 915:923 arg1 folding 847:853 folding 847:853 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 10 41 theme insect 1868:1873 arg1 cells 1889:1893 insect and mammalian cells 1868:1893 insect and mammalian cells 1868:1893 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 6 42 theme functional 891:900 arg1 complexes 915:923 functional glycoprotein complexes 891:923 functional glycoprotein complexes 891:923 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 9 43 theme Sf9 1653:1655 arg1 cells 1657:1661 Sf9 cells 1653:1661 Sf9 cells 1653:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 8 44 theme proteins 1409:1416 arg1 mobility 1390:1397 the electrophoretic mobility 1370:1397 the electrophoretic mobility of mutant proteins 1370:1416 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 45 theme glycoprotein 1265:1276 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 45 theme glycoprotein 1265:1276 arg1 sites 1249:1253 the single glycosylation sites 1224:1253 the single glycosylation sites of the E2 glycoprotein 1224:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 0 46 theme insect 86:91 arg1 cells 107:111 insect and Mammalian cells 86:111 insect and Mammalian cells 86:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 8 47 gly glycosylation 1235:1247 arg2 sites 1249:1253 the single glycosylation sites 1224:1253 the single glycosylation sites of the E2 glycoprotein 1224:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 47 gly glycosylation 1235:1247 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 47 gly glycosylation 1235:1247 arg2 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 5 48 theme glycoproteins 700:712 arg1 N-glycans 687:695 N-glycans 687:695 N-glycans of glycoproteins 687:712 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 11 49 theme RNA 1982:1984 arg1 synthesis 1986:1994 the RNA synthesis 1978:1994 the RNA synthesis of structural proteins 1978:2017 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 1 50 theme particles 240:248 arg1 formation 216:224 the formation 212:224 the formation of infectious particles in infected cells 212:266 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 11 51 theme virus-like 2036:2045 arg1 particles 2047:2055 virus-like particles 2036:2055 virus-like particles in insect and mammalian cells 2036:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 3 52 theme virion 367:372 arg1 processes 443:451 the least understood processes 422:451 the least understood processes 422:451 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 52 theme virion 367:372 arg1 release 387:393 release 387:393 release 387:393 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 52 theme virion 367:372 arg1 assembly 374:381 virion assembly 367:381 virion assembly 367:381 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 7 53 theme 1b 1090:1091 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 53 theme 1b 1090:1091 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 4 54 theme life 675:678 arg1 cycle 680:684 the HCV life cycle 667:684 the HCV life cycle 667:684 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 5 55 theme virus 754:758 arg1 binding 760:766 virus binding 754:766 virus binding to cell surface 754:782 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 1 56 theme infectious 229:238 arg1 particles 240:248 infectious particles 229:248 infectious particles 229:248 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 8 57 theme N6 1294:1295 arg1 site 1297:1300 the N6 site 1290:1300 the N6 site 1290:1300 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 58 gly glycosylation 1475:1487 arg2 sites 1489:1493 glycosylation sites 1475:1493 glycosylation sites 1475:1493 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 11 59 theme proteins 2010:2017 arg1 formation 2023:2031 formation 2023:2031 formation of virus-like particles in insect and mammalian cells 2023:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 59 theme proteins 2010:2017 arg1 synthesis 1986:1994 the RNA synthesis 1978:1994 the RNA synthesis of structural proteins 1978:2017 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 7 60 theme protein 1069:1075 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 60 theme protein 1069:1075 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 0 61 theme e2 16:17 arg1 glycosylation 27:39 HCV e2 protein glycosylation 12:39 HCV e2 protein glycosylation 12:39 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 3 62 dep assembly 374:381 arg1 the 363:365 the 363:365 the 363:365 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 6 63 from folding 847:853 arg1 cells 969:973 insect and mammalian cells 948:973 insect and mammalian cells 948:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 10 64 theme glycans 1697:1703 arg1 elimination 1682:1692 elimination 1682:1692 elimination of glycans at the N1, N2, and N10 sites 1682:1732 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 5 65 theme viral 728:732 arg1 formation 743:751 viral particle formation 728:751 viral particle formation 728:751 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 9 66 from sites 1632:1636 arg1 glycans 1593:1599 glycans 1593:1599 glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells 1593:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 66 from sites 1632:1636 arg1 presence 1581:1588 the presence 1577:1588 the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells 1577:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 11 67 theme particles 2047:2055 arg1 formation 2023:2031 formation 2023:2031 formation of virus-like particles in insect and mammalian cells 2023:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 67 theme particles 2047:2055 arg1 synthesis 1986:1994 the RNA synthesis 1978:1994 the RNA synthesis of structural proteins 1978:2017 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 7 68 theme expression 1190:1199 arg1 system 1201:1206 the baculovirus expression system 1174:1206 the baculovirus expression system 1174:1206 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 6 69 gly glycoprotein 902:913 arg1 glycoprotein 902:913 functional glycoprotein complexes 891:923 functional glycoprotein complexes 891:923 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 2 70 theme detailed 273:280 arg1 structure 282:290 The detailed structure 269:290 The detailed structure of the infectious particle of HCV 269:324 The detailed structure of the infectious particle of HCV remains poorly understood. 25927005 9 71 gly glycoprotein 1526:1537 arg1 glycoprotein 1526:1537 HCV glycoprotein E2 1522:1540 HCV glycoprotein E2 1522:1540 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 72 theme synthesis 1509:1517 arg1 level 1500:1504 The level 1496:1504 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells 1496:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 4 73 theme proteins 538:545 arg1 glycosylation 502:514 glycosylation 502:514 glycosylation of the virus envelope proteins in a cell 502:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 11 74 from particles 2047:2055 arg1 cells 2081:2085 insect and mammalian cells 2060:2085 insect and mammalian cells 2060:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 8 75 theme synthesis 1322:1330 arg1 efficiency 1332:1341 its synthesis efficiency 1318:1341 its synthesis efficiency in Sf9 insect cells 1318:1361 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 9 76 theme HCV 1522:1524 arg1 E2 1539:1540 HCV glycoprotein E2 1522:1540 HCV glycoprotein E2 1522:1540 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 4 77 gly glycosylation 582:594 arg2 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 77 gly glycosylation 582:594 arg2 sites 596:600 glycansat several glycosylation sites 564:600 glycansat several glycosylation sites of these proteins 564:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 77 gly glycosylation 582:594 arg1 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 11 78 from synthesis 1986:1994 arg1 cells 2081:2085 insect and mammalian cells 2060:2085 insect and mammalian cells 2060:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 2 79 theme infectious 299:308 arg1 particle 310:317 the infectious particle 295:317 the infectious particle of HCV 295:324 The detailed structure of the infectious particle of HCV remains poorly understood. 25927005 9 80 theme E2 1539:1540 arg1 synthesis 1509:1517 synthesis 1509:1517 synthesis of HCV glycoprotein E2 in human HEK293T cells 1509:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 11 81 theme HCV 1954:1956 arg1 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 10 82 theme N10 1724:1726 arg1 sites 1728:1732 the N1, N2, and N10 sites 1708:1732 the N1, N2, and N10 sites 1708:1732 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 9 83 theme human 1545:1549 arg1 cells 1559:1563 human HEK293T cells 1545:1563 human HEK293T cells 1545:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 8 84 theme sites 1249:1253 arg1 Elimination 1209:1219 Elimination 1209:1219 Elimination 1209:1219 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 4 85 theme virus 523:527 arg1 proteins 538:545 the virus envelope proteins 519:545 the virus envelope proteins in a cell 519:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 5 86 theme HCV 789:791 arg1 pathogenesis 793:804 HCV pathogenesis 789:804 HCV pathogenesis 789:804 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 0 87 theme glycosylation 27:39 arg1 role 4:7 The role 0:7 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 0:112 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 4 88 theme glycosylation 582:594 arg1 sites 596:600 glycansat several glycosylation sites 564:600 glycansat several glycosylation sites of these proteins 564:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 88 theme glycosylation 582:594 arg1 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 89 from proteins 538:545 arg1 cell 552:555 a cell 550:555 a cell 550:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 1 90 from formation 216:224 arg1 cells 262:266 infected cells 253:266 infected cells 253:266 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 91 theme envelope 142:149 arg1 proteins 151:158 The hepatitis C virus (HCV) envelope proteins 114:158 The hepatitis C virus (HCV) envelope proteins E1 and E2 114:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 91 theme envelope 142:149 arg1 E2 167:168 E2 167:168 E2 167:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 91 theme envelope 142:149 arg1 E1 160:161 E1 160:161 E1 160:161 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 8 92 theme mutant 1402:1407 arg1 proteins 1409:1416 mutant proteins 1402:1416 mutant proteins 1402:1416 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 93 from efficiency 1332:1341 arg1 cells 1357:1361 Sf9 insect cells 1346:1361 Sf9 insect cells 1346:1361 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 6 94 dep folding 847:853 arg1 glycoprotein 864:875 ofthe E2 glycoprotein 855:875 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 10 95 theme E1E2 1774:1777 arg1 dimers 1779:1784 unproductive E1E2 dimers 1761:1784 unproductive E1E2 dimers 1761:1784 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 10 96 theme same 1671:1674 arg1 time 1676:1679 the same time 1667:1679 the same time 1667:1679 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 5 97 theme cell 771:774 arg1 surface 776:782 cell surface 771:782 cell surface 771:782 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 6 98 theme E2 861:862 arg1 glycoprotein 864:875 ofthe E2 glycoprotein 855:875 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 9 99 theme glycans 1593:1599 arg1 presence 1581:1588 the presence 1577:1588 the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells 1577:1661 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 8 100 theme insect 1350:1355 arg1 cells 1357:1361 Sf9 insect cells 1346:1361 Sf9 insect cells 1346:1361 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 4 101 theme glycansat 564:572 arg1 sites 596:600 glycansat several glycosylation sites 564:600 glycansat several glycosylation sites of these proteins 564:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 101 theme glycansat 564:572 arg1 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 10 102 theme assembly 1815:1822 arg1 suppression 1824:1834 productive assembly suppression 1804:1834 productive assembly suppression 1804:1834 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 11 103 theme single 1924:1929 arg1 sites 1945:1949 single glycosylation sites 1924:1949 single glycosylation sites of HCV E2 1924:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 103 theme single 1924:1929 arg1 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 8 104 dep decrease 1449:1456 arg1 proportion 1431:1440 proportion 1431:1440 proportion 1431:1440 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 11 105 from cells 2081:2085 arg1 formation 2023:2031 formation 2023:2031 formation of virus-like particles in insect and mammalian cells 2023:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 105 from cells 2081:2085 arg1 synthesis 1986:1994 the RNA synthesis 1978:1994 the RNA synthesis of structural proteins 1978:2017 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 4 106 gly glycosylation 502:514 arg1 proteins 538:545 the virus envelope proteins 519:545 the virus envelope proteins in a cell 519:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 0 107 theme protein 19:25 arg1 glycosylation 27:39 HCV e2 protein glycosylation 12:39 HCV e2 protein glycosylation 12:39 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 6 108 theme insect 948:953 arg1 cells 969:973 insect and mammalian cells 948:973 insect and mammalian cells 948:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 10 109 theme particles 1850:1858 arg1 aggregates 1789:1798 aggregates 1789:1798 aggregates 1789:1798 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 4 110 from cell 552:555 arg1 glycosylation 502:514 glycosylation 502:514 glycosylation of the virus envelope proteins in a cell 502:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 1 111 theme hepatitis 118:126 arg1 HCV 137:139 HCV 137:139 HCV 137:139 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 111 theme hepatitis 118:126 arg1 virus 130:134 hepatitis C virus 118:134 The hepatitis C virus (HCV) envelope proteins E1 and E2 114:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 9 112 theme glycosylation 1618:1630 arg1 sites 1632:1636 the N1 and N8 glycosylation sites 1604:1636 sites 1632:1636 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 8 113 theme sites 1489:1493 arg1 number 1465:1470 the number 1461:1470 the number of glycosylation sites 1461:1493 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 4 114 theme proteins 611:618 arg1 sites 596:600 glycansat several glycosylation sites 564:600 glycansat several glycosylation sites of these proteins 564:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 4 114 theme proteins 611:618 arg1 proteins 611:618 these proteins 605:618 these proteins 605:618 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 0 115 theme envelope 65:72 arg1 proteins 74:81 virus envelope proteins 59:81 virus envelope proteins in insect and Mammalian cells 59:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 1 116 theme virus 130:134 arg1 proteins 151:158 The hepatitis C virus (HCV) envelope proteins 114:158 The hepatitis C virus (HCV) envelope proteins E1 and E2 114:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 116 theme virus 130:134 arg1 E2 167:168 E2 167:168 E2 167:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 116 theme virus 130:134 arg1 E1 160:161 E1 160:161 E1 160:161 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 0 117 from role 4:7 arg1 functioning 44:54 functioning 44:54 functioning 44:54 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 8 118 theme glycosylation 1235:1247 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 8 118 theme glycosylation 1235:1247 arg1 sites 1249:1253 the single glycosylation sites 1224:1253 the single glycosylation sites of the E2 glycoprotein 1224:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 11 119 theme insect 2060:2065 arg1 cells 2081:2085 insect and mammalian cells 2060:2085 insect and mammalian cells 2060:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 6 120 theme glycoprotein 902:913 arg1 complexes 915:923 functional glycoprotein complexes 891:923 functional glycoprotein complexes 891:923 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 11 121 theme mammalian 2071:2079 arg1 cells 2081:2085 insect and mammalian cells 2060:2085 insect and mammalian cells 2060:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 7 122 theme strain 1093:1098 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 122 theme strain 1093:1098 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 9 123 from presence 1581:1588 arg1 sites 1632:1636 the N1 and N8 glycosylation sites 1604:1636 sites 1632:1636 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 123 from presence 1581:1588 arg1 N1 1608:1609 the N1 and N8 glycosylation sites 1604:1636 N1 1608:1609 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 10 124 dep aggregates 1789:1798 arg1 both 1860:1863 both 1860:1863 both 1860:1863 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 8 125 theme E2 1262:1263 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 5 126 theme particle 734:741 arg1 formation 743:751 viral particle formation 728:751 viral particle formation 728:751 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 8 127 theme glycosylation 1475:1487 arg1 sites 1489:1493 glycosylation sites 1475:1493 glycosylation sites 1475:1493 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 9 128 from synthesis 1509:1517 arg1 cells 1559:1563 human HEK293T cells 1545:1563 human HEK293T cells 1545:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 2 129 theme HCV 322:324 arg1 particle 310:317 the infectious particle 295:317 the infectious particle of HCV 295:324 The detailed structure of the infectious particle of HCV remains poorly understood. 25927005 0 130 theme HCV 12:14 arg1 glycosylation 27:39 HCV e2 protein glycosylation 12:39 HCV e2 protein glycosylation 12:39 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 9 131 from level 1500:1504 arg1 cells 1559:1563 human HEK293T cells 1545:1563 human HEK293T cells 1545:1563 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 9 132 dep cells 1657:1661 arg1 contrast 1641:1648 contrast 1641:1648 contrast 1641:1648 The level of synthesis of HCV glycoprotein E2 in human HEK293T cells depended on the presence of glycans at the N1 and N8 glycosylation sites in contrast to Sf9 cells. 25927005 4 133 theme HCV 671:673 arg1 cycle 680:684 the HCV life cycle 667:684 the HCV life cycle 667:684 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 7 134 theme HCV 1065:1067 arg1 274933RU 1100:1107 genotype 1b strain 274933RU 1081:1107 genotype 1b strain 274933RU 1081:1107 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 7 134 theme HCV 1065:1067 arg1 E2 1077:1078 HCV protein E2 1065:1078 HCV protein E2 (genotype 1b strain 274933RU) 1065:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 8 135 gly glycoprotein 1265:1276 arg1 glycoprotein 1265:1276 the E2 glycoprotein 1258:1276 the E2 glycoprotein 1258:1276 Elimination of the single glycosylation sites of the E2 glycoprotein, except for the N6 site, did not affect its synthesis efficiency in Sf9 insect cells, while the electrophoretic mobility of mutant proteins increased in proportion to the decrease in the number of glycosylation sites. 25927005 11 136 gly glycosylation 1931:1943 arg1 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 136 gly glycosylation 1931:1943 arg2 sites 1945:1949 single glycosylation sites 1924:1949 single glycosylation sites of HCV E2 1924:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 11 136 gly glycosylation 1931:1943 arg2 E2 1958:1959 HCV E2 1954:1959 HCV E2 1954:1959 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 6 137 theme mammalian 959:967 arg1 cells 969:973 insect and mammalian cells 948:973 insect and mammalian cells 948:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 6 138 from formation 878:886 arg1 cells 969:973 insect and mammalian cells 948:973 insect and mammalian cells 948:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 1 139 dep proteins 151:158 arg1 proteins 151:158 The hepatitis C virus (HCV) envelope proteins 114:158 The hepatitis C virus (HCV) envelope proteins E1 and E2 114:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 139 dep proteins 151:158 arg1 E2 167:168 E2 167:168 E2 167:168 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 1 139 dep proteins 151:158 arg1 E1 160:161 E1 160:161 E1 160:161 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 3 140 theme virions 398:404 arg1 processes 443:451 the least understood processes 422:451 the least understood processes 422:451 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 140 theme virions 398:404 arg1 release 387:393 release 387:393 release 387:393 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 3 140 theme virions 398:404 arg1 assembly 374:381 virion assembly 367:381 virion assembly 367:381 Moreover, the virion assembly and release of virions by the cell are the least understood processes. 25927005 7 141 theme sites 1056:1060 arg1 mutations 1023:1031 point mutations 1017:1031 point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) 1017:1108 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 1 142 theme infected 253:260 arg1 cells 262:266 infected cells 253:266 infected cells 253:266 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 7 143 theme baculovirus 1178:1188 arg1 system 1201:1206 the baculovirus expression system 1174:1206 the baculovirus expression system 1174:1206 In order to investigate these processes, point mutations of the N-glycosylation sites of HCV protein E2 (genotype 1b strain 274933RU) were generated and the mutant proteins were further analyzed in the baculovirus expression system. 25927005 11 144 theme structural 1999:2008 arg1 proteins 2010:2017 structural proteins 1999:2017 structural proteins 1999:2017 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 4 145 theme virion 474:479 arg1 properties 481:490 virion properties 474:490 virion properties 474:490 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 25927005 11 146 from formation 2023:2031 arg1 cells 2081:2085 insect and mammalian cells 2060:2085 insect and mammalian cells 2060:2085 In addition, elimination of single glycosylation sites of HCV E2 had no impact on the RNA synthesis of structural proteins and formation of virus-like particles in insect and mammalian cells. 25927005 10 147 theme N2 1716:1717 arg1 sites 1728:1732 the N1, N2, and N10 sites 1708:1732 the N1, N2, and N10 sites 1708:1732 At the same time, elimination of glycans at the N1, N2, and N10 sites led to the accumulation of unproductive E1E2 dimers as aggregates and productive assembly suppression of virus-like particles both in insect and mammalian cells. 25927005 5 148 gly glycoproteins 700:712 arg1 glycoproteins 700:712 glycoproteins 700:712 glycoproteins 700:712 N-glycans of glycoproteins can influence viral particle formation, virus binding to cell surface, and HCV pathogenesis. 25927005 2 149 theme particle 310:317 arg1 structure 282:290 The detailed structure 269:290 The detailed structure of the infectious particle of HCV 269:324 The detailed structure of the infectious particle of HCV remains poorly understood. 25927005 1 150 theme virion 177:182 arg1 components 184:193 virion components 177:193 virion components 177:193 The hepatitis C virus (HCV) envelope proteins E1 and E2, being virion components, are involved in the formation of infectious particles in infected cells. 25927005 0 151 gly glycosylation 27:39 arg1 proteins 74:81 virus envelope proteins 59:81 virus envelope proteins in insect and Mammalian cells 59:111 The role of HCV e2 protein glycosylation in functioning of virus envelope proteins in insect and Mammalian cells. 25927005 6 152 from effect 822:827 arg1 particles 935:943 virus particles 929:943 virus particles in insect and mammalian cells 929:973 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 6 152 from effect 822:827 arg1 formation 878:886 formation 878:886 formation of functional glycoprotein complexes 878:923 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 6 152 from effect 822:827 arg1 folding 847:853 folding 847:853 the folding ofthe E2 glycoprotein 843:875 We studied the effect of glycans on the folding ofthe E2 glycoprotein, formation of functional glycoprotein complexes and virus particles in insect and mammalian cells. 25927005 4 153 from glycosylation 502:514 arg1 cell 552:555 a cell 550:555 a cell 550:555 It is believed that virion properties depend on glycosylation of the virus envelope proteins in a cell, while glycansat several glycosylation sites of these proteins play a pivotal role in protein functioning and the HCV life cycle. 27458127 0 0 theme antibodies 82:91 arg1 profiling 47:55 Fast, robust and high-resolution glycosylation profiling 0:55 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies 0:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 6 1 theme digestion 1095:1103 arg1 steps 1105:1109 no digestion steps 1092:1109 no digestion steps 1092:1109 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 0 2 theme IgG 78:80 arg1 antibodies 82:91 intact monoclonal IgG antibodies 60:91 intact monoclonal IgG antibodies 60:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 5 3 theme free 986:989 arg1 N-glycan 991:998 free N-glycan 986:998 free N-glycan 986:998 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 7 4 with patients 1459:1466 arg1 gammopathies 1484:1495 monoclonal gammopathies 1473:1495 monoclonal gammopathies 1473:1495 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 7 5 theme new 1388:1390 arg1 opportunities 1392:1404 new opportunities 1388:1404 new opportunities 1388:1404 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 5 6 dep lack 893:896 arg1 i.e. 884:887 i.e. 884:887 i.e. 884:887 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 1 7 from generation 178:187 arg1 essential 161:169 essential 161:169 essential 161:169 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 2 8 theme biopharmaceuticals 318:335 arg1 field 309:313 the field 305:313 the field of biopharmaceuticals 305:335 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 27458127 6 9 from information 1166:1176 arg1 site 1217:1220 each IgG attachment site 1197:1220 each IgG attachment site 1197:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 1 10 from essential 161:169 arg1 generation 178:187 the generation 174:187 the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties 174:284 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 3 11 theme sample 619:624 arg1 preparation 626:636 lengthy sample preparation 611:636 lengthy sample preparation with the possibility to introduce artifacts 611:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 7 12 from impact 1428:1433 arg1 patients 1459:1466 patients 1459:1466 patients with monoclonal gammopathies 1459:1495 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 4 13 theme IgG 807:809 arg1 profiling 825:833 intact monoclonal IgG glycosylation profiling 789:833 intact monoclonal IgG glycosylation profiling 789:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 4 14 theme monoclonal 796:805 arg1 profiling 825:833 intact monoclonal IgG glycosylation profiling 789:833 intact monoclonal IgG glycosylation profiling 789:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 3 15 theme mass 496:499 arg1 methods 514:520 proteolysis-based mass spectrometry methods 478:520 proteolysis-based mass spectrometry methods 478:520 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 15 theme mass 496:499 arg1 analyses 428:435 Current analyses 420:435 Current analyses of monoclonal antibody glycosylation 420:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 4 16 theme high-resolution 720:734 arg1 method 753:758 a fast, robust and high-resolution nanoLC-chip-QTOF method 701:758 a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling 701:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 6 17 theme IgG 1202:1204 arg1 site 1217:1220 each IgG attachment site 1197:1220 each IgG attachment site 1197:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 3 18 theme monoclonal 440:449 arg1 antibody 451:458 monoclonal antibody 440:458 monoclonal antibody glycosylation 440:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 1 19 theme therapeutic 192:202 arg1 biologicals 204:214 therapeutic biologicals 192:214 therapeutic biologicals 192:214 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 3 20 with preparation 626:636 arg1 possibility 647:657 the possibility to introduce artifacts 643:680 the possibility to introduce artifacts 643:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 7 21 theme quality 1306:1312 arg1 control 1314:1320 quality control 1306:1320 quality control of the production process of mAb biopharmaceuticals 1306:1372 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 2 22 theme fast 363:366 arg1 analysis 393:400 fast, robust and quantitative analysis 363:400 fast, robust and quantitative analysis of glycosylation 363:417 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 27458127 7 23 theme mAb 1351:1353 arg1 biopharmaceuticals 1355:1372 mAb biopharmaceuticals 1351:1372 mAb biopharmaceuticals 1351:1372 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 1 24 theme biologicals 204:214 arg1 generation 178:187 the generation 174:187 the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties 174:284 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 0 25 theme Fast 0:3 arg1 profiling 47:55 Fast, robust and high-resolution glycosylation profiling 0:55 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies 0:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 5 26 theme important 928:936 arg1 advantage 938:946 an important advantage 925:946 an important advantage over the well-established methods for free N-glycan or glycopeptide analysis 925:1023 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 5 26 theme important 928:936 arg1 hypoglycosylation 865:881 hypoglycosylation 865:881 hypoglycosylation 865:881 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 4 27 theme profiling 825:833 arg1 analysis 777:784 quantitative analysis 764:784 quantitative analysis of intact monoclonal IgG glycosylation profiling 764:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 3 28 theme spectrometry 501:512 arg1 methods 514:520 proteolysis-based mass spectrometry methods 478:520 proteolysis-based mass spectrometry methods 478:520 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 28 theme spectrometry 501:512 arg1 analyses 428:435 Current analyses 420:435 Current analyses of monoclonal antibody glycosylation 420:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 29 theme Current 420:426 arg1 methods 514:520 proteolysis-based mass spectrometry methods 478:520 proteolysis-based mass spectrometry methods 478:520 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 29 theme Current 420:426 arg1 analyses 428:435 Current analyses 420:435 Current analyses of monoclonal antibody glycosylation 420:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 7 30 theme production 1329:1338 arg1 process 1340:1346 the production process 1325:1346 the production process of mAb biopharmaceuticals 1325:1372 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 3 31 theme detailed 537:544 arg1 information 557:567 detailed structural information 537:567 detailed structural information 537:567 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 0 32 theme robust 6:11 arg1 profiling 47:55 Fast, robust and high-resolution glycosylation profiling 0:55 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies 0:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 1 33 gly glycosylation 125:137 arg1 immunoglobulins 142:156 immunoglobulins 142:156 immunoglobulins 142:156 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 1 34 theme Optimal 117:123 arg1 glycosylation 125:137 Optimal glycosylation 117:137 Optimal glycosylation of immunoglobulins 117:156 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 3 35 theme structural 546:555 arg1 information 557:567 detailed structural information 537:567 detailed structural information 537:567 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 0 36 theme glycosylation 33:45 arg1 profiling 47:55 Fast, robust and high-resolution glycosylation profiling 0:55 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies 0:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 4 37 theme intact 789:794 arg1 profiling 825:833 intact monoclonal IgG glycosylation profiling 789:833 intact monoclonal IgG glycosylation profiling 789:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 6 38 theme attachment 1206:1215 arg1 site 1217:1220 each IgG attachment site 1197:1220 each IgG attachment site 1197:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 7 39 theme monoclonal 1473:1482 arg1 gammopathies 1484:1495 monoclonal gammopathies 1473:1495 monoclonal gammopathies 1473:1495 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 2 40 from challenge 292:300 arg1 field 309:313 the field 305:313 the field of biopharmaceuticals 305:335 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 27458127 4 41 theme robust 709:714 arg1 method 753:758 a fast, robust and high-resolution nanoLC-chip-QTOF method 701:758 a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling 701:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 0 42 theme high-resolution 17:31 arg1 profiling 47:55 Fast, robust and high-resolution glycosylation profiling 0:55 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies 0:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 3 43 theme proteolysis-based 478:494 arg1 methods 514:520 proteolysis-based mass spectrometry methods 478:520 proteolysis-based mass spectrometry methods 478:520 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 43 theme proteolysis-based 478:494 arg1 analyses 428:435 Current analyses 420:435 Current analyses of monoclonal antibody glycosylation 420:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 5 44 theme whole 901:905 arg1 glycans 907:913 whole glycans 901:913 whole glycans 901:913 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 5 45 gly glycopeptide 1003:1014 arg2 glycopeptide 1003:1014 glycopeptide analysis 1003:1023 glycopeptide analysis 1003:1023 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 6 46 theme relative 1144:1151 arg1 information 1166:1176 direct relative quantitative information 1137:1176 direct relative quantitative information of both glycans on each IgG attachment site 1137:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 3 47 theme antibody 451:458 arg1 glycosylation 460:472 monoclonal antibody glycosylation 440:472 monoclonal antibody glycosylation 440:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 5 48 theme well-established 957:972 arg1 methods 974:980 the well-established methods 953:980 the well-established methods 953:980 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 3 49 dep methods 514:520 arg1 provide 529:535 provide 529:535 provide detailed structural information 529:567 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 49 dep methods 514:520 arg1 suffer 574:579 suffer 574:579 suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts 574:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 2 50 theme quantitative 380:391 arg1 analysis 393:400 fast, robust and quantitative analysis 363:400 fast, robust and quantitative analysis of glycosylation 363:417 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 27458127 3 51 theme glycosylation 460:472 arg1 methods 514:520 proteolysis-based mass spectrometry methods 478:520 proteolysis-based mass spectrometry methods 478:520 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 51 theme glycosylation 460:472 arg1 analyses 428:435 Current analyses 420:435 Current analyses of monoclonal antibody glycosylation 420:472 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 6 52 theme direct 1137:1142 arg1 information 1166:1176 direct relative quantitative information 1137:1176 direct relative quantitative information of both glycans on each IgG attachment site 1137:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 7 53 theme biopharmaceuticals 1355:1372 arg1 process 1340:1346 the production process 1325:1346 the production process of mAb biopharmaceuticals 1325:1372 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 4 54 theme nanoLC-chip-QTOF 736:751 arg1 method 753:758 a fast, robust and high-resolution nanoLC-chip-QTOF method 701:758 a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling 701:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 4 55 theme glycosylation 811:823 arg1 profiling 825:833 intact monoclonal IgG glycosylation profiling 789:833 intact monoclonal IgG glycosylation profiling 789:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 7 56 theme process 1340:1346 arg1 control 1314:1320 quality control 1306:1320 quality control of the production process of mAb biopharmaceuticals 1306:1372 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 7 57 theme mAb-glycosylation 1438:1454 arg1 impact 1428:1433 the clinical impact 1415:1433 the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies 1415:1495 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 7 58 theme clinical 1419:1426 arg1 impact 1428:1433 the clinical impact 1415:1433 the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies 1415:1495 We demonstrate that the ease and robustness make this technique ideally suited for quality control of the production process of mAb biopharmaceuticals, and provides new opportunities to study the clinical impact of mAb-glycosylation in patients with monoclonal gammopathies. 27458127 1 59 theme immunoglobulins 142:156 arg1 glycosylation 125:137 Optimal glycosylation 117:137 Optimal glycosylation of immunoglobulins 117:156 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 5 60 theme glycopeptide 1003:1014 arg1 analysis 1016:1023 glycopeptide analysis 1003:1023 glycopeptide analysis 1003:1023 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 3 61 theme lengthy 611:617 arg1 preparation 626:636 lengthy sample preparation 611:636 lengthy sample preparation with the possibility to introduce artifacts 611:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 62 theme drawbacks 593:601 arg1 number 583:588 a number 581:588 a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts 581:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 3 62 theme drawbacks 593:601 arg1 preparation 626:636 lengthy sample preparation 611:636 lengthy sample preparation with the possibility to introduce artifacts 611:680 Current analyses of monoclonal antibody glycosylation are proteolysis-based mass spectrometry methods, which provide detailed structural information, but suffer a number of drawbacks such as lengthy sample preparation with the possibility to introduce artifacts. 27458127 2 63 theme robust 369:374 arg1 analysis 393:400 fast, robust and quantitative analysis 363:400 fast, robust and quantitative analysis of glycosylation 363:417 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 27458127 0 64 theme monoclonal 67:76 arg1 antibodies 82:91 intact monoclonal IgG antibodies 60:91 intact monoclonal IgG antibodies 60:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 6 65 theme glycans 1186:1192 arg1 information 1166:1176 direct relative quantitative information 1137:1176 direct relative quantitative information of both glycans on each IgG attachment site 1137:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 4 66 theme fast 703:706 arg1 method 753:758 a fast, robust and high-resolution nanoLC-chip-QTOF method 701:758 a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling 701:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 4 67 theme quantitative 764:775 arg1 analysis 777:784 quantitative analysis 764:784 quantitative analysis of intact monoclonal IgG glycosylation profiling 764:833 Here, we describe a fast, robust and high-resolution nanoLC-chip-QTOF method for quantitative analysis of intact monoclonal IgG glycosylation profiling. 27458127 6 68 theme quantitative 1153:1164 arg1 information 1166:1176 direct relative quantitative information 1137:1176 direct relative quantitative information of both glycans on each IgG attachment site 1137:1220 Moreover, the method is highly amenable to automation and because no digestion steps are involved, it provides direct relative quantitative information of both glycans on each IgG attachment site. 27458127 1 69 theme immunogenic 263:273 arg1 properties 275:284 immunogenic properties 263:284 immunogenic properties 263:284 Optimal glycosylation of immunoglobulins is essential in the generation of therapeutic biologicals with respect to efficacy, pharmacokinetics and immunogenic properties. 27458127 0 70 theme intact 60:65 arg1 antibodies 82:91 intact monoclonal IgG antibodies 60:91 intact monoclonal IgG antibodies 60:91 Fast, robust and high-resolution glycosylation profiling of intact monoclonal IgG antibodies using nanoLC-chip-QTOF. 27458127 5 71 theme glycans 907:913 arg1 lack 893:896 the lack 889:896 the lack of whole glycans 889:913 The method is able to detect hypoglycosylation, i.e. the lack of whole glycans, which is an important advantage over the well-established methods for free N-glycan or glycopeptide analysis. 27458127 2 72 theme glycosylation 405:417 arg1 analysis 393:400 fast, robust and quantitative analysis 363:400 fast, robust and quantitative analysis of glycosylation 363:417 This challenge in the field of biopharmaceuticals requires technologies for fast, robust and quantitative analysis of glycosylation. 25160934 9 0 dep control 1767:1773 arg1 the 1763:1765 the 1763:1765 the 1763:1765 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 2 1 theme domain 616:621 arg1 features 623:630 the variable and constant domain features 590:630 the variable and constant domain features of the antibodies 590:648 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 2 theme constant 607:614 arg1 features 623:630 the variable and constant domain features 590:630 the variable and constant domain features of the antibodies 590:648 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 3 theme Fc-effector 668:678 arg1 functions 680:688 robust Fc-effector functions 661:688 robust Fc-effector functions 661:688 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 9 4 theme emerging 1717:1724 arg1 function 1738:1745 an emerging Fc-effector function 1714:1745 an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1714:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 9 4 theme emerging 1717:1724 arg1 activity 1704:1711 enhanced ADCP activity 1690:1711 enhanced ADCP activity 1690:1711 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 4 5 theme glycan 946:951 arg1 profiles 953:960 Antibody glycan profiles 937:960 Antibody glycan profiles 937:960 Antibody glycan profiles were determined by HPLC. 25160934 8 6 theme Fc-effector 1553:1563 arg1 functions 1565:1573 Fc-effector functions 1553:1573 Fc-effector functions 1553:1573 CONCLUSIONS: Our studies point to the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity. 25160934 7 7 theme independent 1413:1423 arg1 FcγRIIIa 1385:1392 FcγRIIIa 1385:1392 FcγRIIIa 1385:1392 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 2 8 theme robust 661:666 arg1 functions 680:688 robust Fc-effector functions 661:688 robust Fc-effector functions 661:688 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 9 theme hamster 300:306 arg1 cells 314:318 Chinese hamster ovary cells 292:318 Chinese hamster ovary cells 292:318 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 6 10 theme decreased 1142:1150 arg1 content 1162:1168 decreased galactose content 1142:1168 decreased galactose content 1142:1168 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 25160934 3 11 theme antibody-dependent 817:834 arg1 ADCP 859:862 ADCP 859:862 ADCP 859:862 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 11 theme antibody-dependent 817:834 arg1 phagocytosis 845:856 antibody-dependent cellular phagocytosis 817:856 antibody-dependent cellular phagocytosis (ADCP) 817:863 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 11 theme antibody-dependent 817:834 arg1 gp140 754:758 gp140(SR162) 754:765 gp140(SR162) 754:765 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 4 12 theme Antibody 937:944 arg1 profiles 953:960 Antibody glycan profiles 937:960 Antibody glycan profiles 937:960 Antibody glycan profiles were determined by HPLC. 25160934 5 13 theme function 1087:1094 arg1 potency 1064:1070 the potency 1060:1070 the potency of Fc-effector function 1060:1094 RESULTS: Neither the specificity nor the affinity of the mAbs determined the potency of Fc-effector function. 25160934 2 14 theme variable 594:601 arg1 features 623:630 the variable and constant domain features 590:630 the variable and constant domain features of the antibodies 590:648 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 15 theme Chinese 292:298 arg1 cells 314:318 Chinese hamster ovary cells 292:318 Chinese hamster ovary cells 292:318 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 3 16 theme cellular 836:843 arg1 ADCP 859:862 ADCP 859:862 ADCP 859:862 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 16 theme cellular 836:843 arg1 phagocytosis 845:856 antibody-dependent cellular phagocytosis 817:856 antibody-dependent cellular phagocytosis (ADCP) 817:863 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 16 theme cellular 836:843 arg1 gp140 754:758 gp140(SR162) 754:765 gp140(SR162) 754:765 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 2 17 theme antibodies 639:648 arg1 features 623:630 the variable and constant domain features 590:630 the variable and constant domain features of the antibodies 590:648 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 18 theme HIV 416:418 arg1 envelope 420:427 the HIV envelope 412:427 the HIV envelope 412:427 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 8 19 theme antibody 1596:1603 arg1 specificity 1605:1615 the antibody specificity 1592:1615 the antibody specificity 1592:1615 CONCLUSIONS: Our studies point to the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity. 25160934 3 20 theme antibody-dependent 768:785 arg1 gp140 754:758 gp140(SR162) 754:765 gp140(SR162) 754:765 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 20 theme antibody-dependent 768:785 arg1 ADCC 810:813 ADCC 810:813 ADCC 810:813 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 20 theme antibody-dependent 768:785 arg1 cytotoxicity 796:807 antibody-dependent cellular cytotoxicity 768:807 antibody-dependent cellular cytotoxicity (ADCC) 768:814 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 7 21 theme mAb 1451:1453 arg1 specificity 1432:1442 the specificity 1428:1442 the specificity of the mAb 1428:1453 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 2 22 theme gp120-CD4 464:472 arg1 site 485:488 gp120-CD4(+) binding site 464:488 gp120-CD4(+) binding site 464:488 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 3 23 theme antibody-binding 725:740 arg1 affinity 742:749 antibody-binding affinity 725:749 antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) 725:863 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 2 24 theme gp41-specific 495:507 arg1 antibodies 509:518 gp41-specific antibodies 495:518 gp41-specific antibodies 495:518 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 25 theme Epstein-Barr 323:334 arg1 heteromyelomas 361:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 7 26 theme enhanced 1365:1372 arg1 binding 1374:1380 enhanced binding 1365:1380 enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb 1365:1453 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 2 27 theme features 623:630 arg1 contribution 574:585 the relative contribution 561:585 the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions 561:688 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 9 28 theme Fc-effector 1726:1736 arg1 function 1738:1745 an emerging Fc-effector function 1714:1745 an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1714:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 9 28 theme Fc-effector 1726:1736 arg1 activity 1704:1711 enhanced ADCP activity 1690:1711 enhanced ADCP activity 1690:1711 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 0 29 theme glycosylation 27:39 arg1 structures 41:50 antibody glycosylation structures 18:50 antibody glycosylation structures that predict monoclonal antibody Fc-effector function 18:104 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 6 30 theme N-glycolylneuraminic 1210:1229 arg1 structures 1247:1256 N-glycolylneuraminic acid-containing structures 1210:1256 N-glycolylneuraminic acid-containing structures 1210:1256 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 25160934 9 31 theme enhanced 1690:1697 arg1 function 1738:1745 an emerging Fc-effector function 1714:1745 an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1714:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 9 31 theme enhanced 1690:1697 arg1 activity 1704:1711 enhanced ADCP activity 1690:1711 enhanced ADCP activity 1690:1711 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 6 32 theme galactose 1152:1160 arg1 content 1162:1168 decreased galactose content 1142:1168 decreased galactose content 1142:1168 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 25160934 2 33 theme gp120-V3 449:456 arg1 loop 458:461 gp120-V3 loop 449:461 gp120-V3 loop 449:461 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 34 theme ovary 308:312 arg1 cells 314:318 Chinese hamster ovary cells 292:318 Chinese hamster ovary cells 292:318 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 1 35 theme -mediated 206:214 arg1 functions 225:233 fragment crystalizable (Fc)-mediated effector functions 179:233 fragment crystalizable (Fc)-mediated effector functions 179:233 OBJECTIVE: To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. 25160934 2 36 theme binding 477:483 arg1 site 485:488 gp120-CD4(+) binding site 464:488 gp120-CD4(+) binding site 464:488 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 1 37 theme effector 216:223 arg1 functions 225:233 fragment crystalizable (Fc)-mediated effector functions 179:233 fragment crystalizable (Fc)-mediated effector functions 179:233 OBJECTIVE: To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. 25160934 0 38 theme structures 41:50 arg1 Identification 0:13 Identification 0:13 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 0:105 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 3 39 theme cellular 787:794 arg1 gp140 754:758 gp140(SR162) 754:765 gp140(SR162) 754:765 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 39 theme cellular 787:794 arg1 ADCC 810:813 ADCC 810:813 ADCC 810:813 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 3 39 theme cellular 787:794 arg1 cytotoxicity 796:807 antibody-dependent cellular cytotoxicity 768:807 antibody-dependent cellular cytotoxicity (ADCC) 768:814 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 8 40 theme specific 1494:1501 arg1 structures 1513:1522 the specific Fc-glycan structures 1490:1522 the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity 1490:1615 CONCLUSIONS: Our studies point to the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity. 25160934 7 41 theme specificity 1432:1442 arg1 independent 1413:1423 independent 1413:1423 independent 1413:1423 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 2 42 theme envelope 420:427 arg1 regions 401:407 key regions 397:407 key regions of the HIV envelope 397:427 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 0 43 theme monoclonal 65:74 arg1 function 97:104 monoclonal antibody Fc-effector function 65:104 monoclonal antibody Fc-effector function 65:104 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 9 44 theme HIV 1792:1794 arg1 infection 1796:1804 HIV infection 1792:1804 HIV infection 1792:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 5 45 theme Fc-effector 1075:1085 arg1 function 1087:1094 Fc-effector function 1075:1094 Fc-effector function 1075:1094 RESULTS: Neither the specificity nor the affinity of the mAbs determined the potency of Fc-effector function. 25160934 6 46 theme enhanced 1268:1275 arg1 ADCP 1277:1280 enhanced ADCP 1268:1280 enhanced ADCP 1268:1280 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 25160934 3 47 dep FcγRIIa 896:902 arg1 receptors 926:934 receptors 926:934 receptors 926:934 METHODS: Each mAb was assayed for antibody-binding affinity to gp140(SR162), antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcγRIIa, FcγRIIb and FcγRIIIa receptors. 25160934 1 48 theme monoclonal 131:140 arg1 mAb 152:154 mAb 152:154 mAb 152:154 OBJECTIVE: To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. 25160934 1 48 theme monoclonal 131:140 arg1 antibody 142:149 monoclonal antibody 131:149 monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV 131:245 OBJECTIVE: To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. 25160934 2 49 attach derived 279:285 arg1 heteromyelomas 361:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 49 attach derived 279:285 arg1 cells 314:318 Chinese hamster ovary cells 292:318 Chinese hamster ovary cells 292:318 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 49 attach derived 279:285 arg2 antibodies 267:276 Monoclonal antibodies 256:276 Monoclonal antibodies 256:276 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 9 50 theme infection 1796:1804 arg1 clearance 1779:1787 clearance 1779:1787 clearance 1779:1787 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 9 50 theme infection 1796:1804 arg1 control 1767:1773 control 1767:1773 control 1767:1773 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 7 51 dep FcγRIIIa 1385:1392 arg1 activity 1403:1410 activity 1403:1410 activity 1403:1410 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 1 52 theme antibody 142:149 arg1 features 157:164 monoclonal antibody (mAb) features 131:164 monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV 131:245 OBJECTIVE: To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. 25160934 2 53 with antibodies 267:276 arg1 specificity 382:392 specificity 382:392 specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies 382:518 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 8 54 theme Fc-glycan 1503:1511 arg1 structures 1513:1522 the specific Fc-glycan structures 1490:1522 the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity 1490:1615 CONCLUSIONS: Our studies point to the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity. 25160934 9 55 theme ADCP 1699:1702 arg1 function 1738:1745 an emerging Fc-effector function 1714:1745 an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1714:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 9 55 theme ADCP 1699:1702 arg1 activity 1704:1711 enhanced ADCP activity 1690:1711 enhanced ADCP activity 1690:1711 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 7 56 theme bi-antenary 1301:1311 arg1 arm 1320:1322 the bi-antenary glycan arm 1297:1322 the bi-antenary glycan arm onto which galactose was added 1297:1353 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 2 57 theme mouse 355:359 arg1 heteromyelomas 361:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 9 58 theme antibody 1647:1654 arg1 structures 1663:1672 antibody glycan structures 1647:1672 antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1647:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 2 59 theme relative 565:572 arg1 contribution 574:585 the relative contribution 561:585 the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions 561:688 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 5 60 theme mAbs 1044:1047 arg1 specificity 1008:1018 the specificity 1004:1018 the specificity 1004:1018 RESULTS: Neither the specificity nor the affinity of the mAbs determined the potency of Fc-effector function. 25160934 6 61 theme acid-containing 1231:1245 arg1 structures 1247:1256 N-glycolylneuraminic acid-containing structures 1210:1256 N-glycolylneuraminic acid-containing structures 1210:1256 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 25160934 2 62 theme virus-immortalized 336:353 arg1 heteromyelomas 361:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 Epstein-Barr virus-immortalized mouse heteromyelomas 323:374 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 7 63 theme glycan 1313:1318 arg1 arm 1320:1322 the bi-antenary glycan arm 1297:1322 the bi-antenary glycan arm onto which galactose was added 1297:1353 Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcγRIIIa and ADCC activity, independent of the specificity of the mAb. 25160934 0 64 theme Fc-effector 85:95 arg1 function 97:104 monoclonal antibody Fc-effector function 65:104 monoclonal antibody Fc-effector function 65:104 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 2 65 theme Monoclonal 256:265 arg1 antibodies 267:276 Monoclonal antibodies 256:276 Monoclonal antibodies 256:276 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 2 66 theme key 397:399 arg1 regions 401:407 key regions 397:407 key regions of the HIV envelope 397:427 DESIGN: Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein-Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4(+) binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. 25160934 9 67 theme glycan 1656:1661 arg1 structures 1663:1672 antibody glycan structures 1647:1672 antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection 1647:1804 Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection. 25160934 0 68 theme antibody 76:83 arg1 function 97:104 monoclonal antibody Fc-effector function 65:104 monoclonal antibody Fc-effector function 65:104 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 0 69 theme antibody 18:25 arg1 structures 41:50 antibody glycosylation structures 18:50 antibody glycosylation structures that predict monoclonal antibody Fc-effector function 18:104 Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function. 25160934 6 70 theme FcγRIIIa 1097:1104 arg1 binding 1106:1112 FcγRIIIa binding 1097:1112 FcγRIIIa binding 1097:1112 FcγRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. 29187599 4 0 theme native 562:567 arg1 hDAO 569:572 native hDAO 562:572 native hDAO from amniotic fluid and Caco-2 cells 562:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 11 1 theme dimer 1688:1692 arg1 formation 1694:1702 efficient DAO dimer formation 1674:1702 efficient DAO dimer formation 1674:1702 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 7 2 theme adjacent 1135:1142 arg1 residues 1126:1133 the residues 1122:1133 the residues adjacent to Asn-110 and Asn-137 1122:1165 X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. 29187599 4 3 from kidneys 629:635 arg1 rhDAO 642:646 rhDAO 642:646 rhDAO produced in two different HEK293 cell lines 642:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 3 from kidneys 629:635 arg1 Asn-110 551:557 Asn-110 551:557 Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines 551:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 3 from kidneys 629:635 arg1 hDAO 569:572 native hDAO 562:572 native hDAO from amniotic fluid and Caco-2 cells 562:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 5 4 theme complex-type 873:884 arg1 glycans 886:892 similar complex-type glycans 865:892 similar complex-type glycans 865:892 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 12 5 theme glycosylation 1794:1806 arg1 sites 1808:1812 all four glycosylation sites 1785:1812 all four glycosylation sites 1785:1812 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 11 6 theme efficient 1674:1682 arg1 formation 1694:1702 efficient DAO dimer formation 1674:1702 efficient DAO dimer formation 1674:1702 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 11 7 theme locations 1584:1592 arg1 important 1660:1668 important 1660:1668 important 1660:1668 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 3 8 theme oligomannosidic 507:521 arg1 glycans 523:529 only mammalian-atypical oligomannosidic glycans 483:529 only mammalian-atypical oligomannosidic glycans 483:529 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 12 9 theme high 1751:1754 arg1 conservation 1769:1780 the high evolutionary conservation 1747:1780 the high evolutionary conservation of all four glycosylation sites 1747:1812 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 4 10 theme Caco-2 598:603 arg1 cells 605:609 Caco-2 cells 598:609 Caco-2 cells 598:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 2 11 gly N-glycosylation 238:252 arg2 sites 254:258 multiple N-glycosylation sites 229:258 multiple N-glycosylation sites 229:258 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 7 12 theme X-ray 1091:1095 arg1 structures 1097:1106 X-ray structures 1091:1106 X-ray structures 1091:1106 X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. 29187599 5 13 theme tetra-antennary 811:825 arg1 branches 827:834 tetra-antennary branches 811:834 tetra-antennary branches 811:834 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 2 14 theme histamine 203:211 arg1 catabolism 213:222 histamine catabolism 203:222 histamine catabolism 203:222 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 4 15 theme HEK293 674:679 arg1 lines 686:690 two different HEK293 cell lines 660:690 two different HEK293 cell lines 660:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 13 16 from abundant 1828:1835 arg1 kidney 1873:1878 kidney 1873:1878 kidney 1873:1878 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 13 16 from abundant 1828:1835 arg1 placenta 1885:1892 placenta 1885:1892 placenta 1885:1892 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 13 16 from abundant 1828:1835 arg1 tract 1866:1870 the gastrointestinal tract 1845:1870 the gastrointestinal tract 1845:1870 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 1 17 theme many 133:136 arg1 processes 149:157 many biological processes 133:157 many biological processes 133:157 N-Glycosylation plays a fundamental role in many biological processes. 29187599 8 18 theme endoplasmic 1344:1354 arg1 reticulum 1356:1364 the endoplasmic reticulum 1340:1364 the endoplasmic reticulum 1340:1364 Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. 29187599 5 19 contain had 861:863 arg2 glycans 886:892 similar complex-type glycans 865:892 similar complex-type glycans 865:892 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 5 19 contain had 861:863 arg1 Asn-745 853:859 Asn-745 853:859 Asn-745 853:859 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 5 19 contain had 861:863 arg1 Asn-538 841:847 Asn-538 841:847 Asn-538 841:847 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 13 20 theme enzyme 1947:1952 arg1 levels 1965:1970 high enzyme expression levels 1942:1970 high enzyme expression levels 1942:1970 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 2 21 theme multiple 229:236 arg1 sites 254:258 multiple N-glycosylation sites 229:258 multiple N-glycosylation sites 229:258 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 2 22 from example 282:288 arg1 secretion 297:305 DAO secretion 293:305 DAO secretion 293:305 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 6 23 theme vascular 997:1004 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 2 24 theme Human 160:164 arg1 hDAO 183:186 hDAO 183:186 hDAO 183:186 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 2 24 theme Human 160:164 arg1 oxidase 174:180 Human diamine oxidase 160:180 Human diamine oxidase (hDAO) 160:187 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 0 25 theme diamine 72:78 arg1 oxidase 80:86 human diamine oxidase 66:86 human diamine oxidase 66:86 Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase. 29187599 5 26 theme similar 865:871 arg1 glycans 886:892 similar complex-type glycans 865:892 similar complex-type glycans 865:892 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 6 27 theme oxidase 983:989 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 3 28 contain carries 475:481 arg1 Asn-110 467:473 Asn-110 467:473 Asn-110 467:473 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 28 contain carries 475:481 arg2 glycans 523:529 only mammalian-atypical oligomannosidic glycans 483:529 only mammalian-atypical oligomannosidic glycans 483:529 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 7 29 theme core 1230:1233 arg1 trisaccharide 1235:1247 the core trisaccharide 1226:1247 the core trisaccharide 1226:1247 X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. 29187599 6 30 theme copper-containing 959:975 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 3 31 contain carry 431:435 arg1 sites 367:371 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 347:401 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) 347:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 31 contain carry 431:435 arg1 Asn-538 382:388 Asn-538 382:388 Asn-538 382:388 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 31 contain carry 431:435 arg1 Asn-745 395:401 Asn-745 395:401 Asn-745 395:401 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 31 contain carry 431:435 arg1 Asn-168 373:379 Asn-168 373:379 Asn-168 373:379 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 31 contain carry 431:435 arg2 glycans 450:456 complex-type glycans 437:456 complex-type glycans 437:456 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 4 32 theme oligomannosidic 725:739 arg1 glycans 741:747 oligomannosidic glycans 725:747 oligomannosidic glycans 725:747 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 5 33 from Asn-168 761:767 arg1 Glycans 750:756 Glycans 750:756 Glycans at Asn-168 750:767 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 3 34 gly N-glycosylation 351:365 arg2 Asn-538 382:388 Asn-538 382:388 Asn-538 382:388 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 34 gly N-glycosylation 351:365 arg2 sites 367:371 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 347:401 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) 347:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 34 gly N-glycosylation 351:365 arg2 Asn-168 373:379 Asn-168 373:379 Asn-168 373:379 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 34 gly N-glycosylation 351:365 arg2 Asn-745 395:401 Asn-745 395:401 Asn-745 395:401 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 35 theme N-glycosylation 351:365 arg1 sites 367:371 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 347:401 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) 347:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 35 theme N-glycosylation 351:365 arg1 Asn-538 382:388 Asn-538 382:388 Asn-538 382:388 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 35 theme N-glycosylation 351:365 arg1 Asn-745 395:401 Asn-745 395:401 Asn-745 395:401 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 35 theme N-glycosylation 351:365 arg1 Asn-168 373:379 Asn-168 373:379 Asn-168 373:379 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 10 36 theme double 1480:1485 arg1 substitutions 1514:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions reduced rhDAO secretion by 85 and 94%. 29187599 0 37 theme human 66:70 arg1 oxidase 80:86 human diamine oxidase 66:86 human diamine oxidase 66:86 Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase. 29187599 0 38 theme Oligomannosidic 0:14 arg1 glycans 16:22 Oligomannosidic glycans 0:22 Oligomannosidic glycans at Asn-110 0:33 Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase. 29187599 9 39 theme Asn-168 1380:1386 arg1 Mutations 1367:1375 Mutations 1367:1375 Mutations of Asn-168, Asn-538, and Asn-745 1367:1408 Mutations of Asn-168, Asn-538, and Asn-745 reduced rhDAO secretion by 13, 71, and 32%, respectively. 29187599 10 40 theme rhDAO 1536:1540 arg1 secretion 1542:1550 rhDAO secretion 1536:1550 rhDAO secretion 1536:1550 Asn-538/745 double and Asn-168/538/745 triple substitutions reduced rhDAO secretion by 85 and 94%. 29187599 7 41 theme conserved 1181:1189 arg1 cleft 1203:1207 a highly conserved hydrophobic cleft 1172:1207 a highly conserved hydrophobic cleft interacting with the core trisaccharide 1172:1247 X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. 29187599 9 42 theme Asn-538 1389:1395 arg1 Mutations 1367:1375 Mutations 1367:1375 Mutations of Asn-168, Asn-538, and Asn-745 1367:1408 Mutations of Asn-168, Asn-538, and Asn-745 reduced rhDAO secretion by 13, 71, and 32%, respectively. 29187599 5 43 theme line-specific 921:933 arg1 variations 935:944 some tissue- and cell line-specific variations 899:944 variations 935:944 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 6 44 theme high-mannose 1052:1063 arg1 glycosylation 1065:1077 high-mannose glycosylation 1052:1077 high-mannose glycosylation 1052:1077 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 13 45 from kidney 1873:1878 arg1 abundant 1828:1835 abundant 1828:1835 abundant 1828:1835 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 2 46 theme diamine 166:172 arg1 hDAO 183:186 hDAO 183:186 hDAO 183:186 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 2 46 theme diamine 166:172 arg1 oxidase 174:180 Human diamine oxidase 160:180 Human diamine oxidase (hDAO) 160:187 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 5 47 gly sialylated 788:797 arg1 Glycans 750:756 Glycans 750:756 Glycans at Asn-168 750:767 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 13 48 from tract 1866:1870 arg1 abundant 1828:1835 abundant 1828:1835 abundant 1828:1835 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 13 49 theme expression 1954:1963 arg1 levels 1965:1970 high enzyme expression levels 1942:1970 high enzyme expression levels 1942:1970 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 11 50 theme DAO 1601:1603 arg1 structure 1605:1613 the DAO structure 1597:1613 the DAO structure 1597:1613 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 4 51 used occupied 713:720 arg2 Asn-110 551:557 Asn-110 551:557 Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines 551:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 52 theme amniotic 579:586 arg1 fluid 588:592 amniotic fluid 579:592 amniotic fluid 579:592 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 12 53 theme sites 1808:1812 arg1 conservation 1769:1780 the high evolutionary conservation 1747:1780 the high evolutionary conservation of all four glycosylation sites 1747:1812 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 3 54 theme mammalian-atypical 488:505 arg1 glycans 523:529 only mammalian-atypical oligomannosidic glycans 483:529 only mammalian-atypical oligomannosidic glycans 483:529 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 12 55 theme evolutionary 1756:1767 arg1 conservation 1769:1780 the high evolutionary conservation 1747:1780 the high evolutionary conservation of all four glycosylation sites 1747:1812 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 11 56 theme DAO 1684:1686 arg1 formation 1694:1702 efficient DAO dimer formation 1674:1702 efficient DAO dimer formation 1674:1702 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 2 57 theme DAO 293:295 arg1 secretion 297:305 DAO secretion 293:305 DAO secretion 293:305 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 6 58 theme related 951:957 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 4 59 theme porcine 621:627 arg1 kidneys 629:635 porcine kidneys 621:635 porcine kidneys 621:635 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 12 60 gly glycosylation 1794:1806 arg2 sites 1808:1812 all four glycosylation sites 1785:1812 all four glycosylation sites 1785:1812 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 12 60 gly glycosylation 1794:1806 arg2 four 1789:1792 four 1789:1792 four 1789:1792 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 4 61 from cells 605:609 arg1 rhDAO 642:646 rhDAO 642:646 rhDAO produced in two different HEK293 cell lines 642:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 61 from cells 605:609 arg1 Asn-110 551:557 Asn-110 551:557 Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines 551:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 61 from cells 605:609 arg1 hDAO 569:572 native hDAO 562:572 native hDAO from amniotic fluid and Caco-2 cells 562:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 8 62 theme rhDAO 1300:1304 arg1 secretion 1306:1314 rhDAO secretion 1300:1314 rhDAO secretion 1300:1314 Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. 29187599 1 63 theme fundamental 113:123 arg1 role 125:128 a fundamental role 111:128 a fundamental role 111:128 N-Glycosylation plays a fundamental role in many biological processes. 29187599 8 64 theme Asn-110 1250:1256 arg1 replacement 1258:1268 Asn-110 replacement 1250:1268 Asn-110 replacement with Gln 1250:1277 Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. 29187599 11 65 from locations 1584:1592 arg1 structure 1605:1613 the DAO structure 1597:1613 the DAO structure 1597:1613 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 8 66 with replacement 1258:1268 arg1 Gln 1275:1277 Gln 1275:1277 Gln 1275:1277 Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. 29187599 4 67 theme different 664:672 arg1 lines 686:690 two different HEK293 cell lines 660:690 two different HEK293 cell lines 660:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 13 68 theme high 1942:1945 arg1 levels 1965:1970 high enzyme expression levels 1942:1970 high enzyme expression levels 1942:1970 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 4 69 from fluid 588:592 arg1 rhDAO 642:646 rhDAO 642:646 rhDAO produced in two different HEK293 cell lines 642:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 69 from fluid 588:592 arg1 Asn-110 551:557 Asn-110 551:557 Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines 551:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 69 from fluid 588:592 arg1 hDAO 569:572 native hDAO 562:572 native hDAO from amniotic fluid and Caco-2 cells 562:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 2 70 theme N-glycosylation 238:252 arg1 sites 254:258 multiple N-glycosylation sites 229:258 multiple N-glycosylation sites 229:258 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 1 71 theme biological 138:147 arg1 processes 149:157 many biological processes 133:157 many biological processes 133:157 N-Glycosylation plays a fundamental role in many biological processes. 29187599 6 72 theme adhesion 1006:1013 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 0 73 from Asn-110 27:33 arg1 glycans 16:22 Oligomannosidic glycans 0:22 Oligomannosidic glycans at Asn-110 0:33 Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase. 29187599 0 74 theme oxidase 80:86 arg1 secretion 53:61 secretion 53:61 secretion of human diamine oxidase 53:86 Oligomannosidic glycans at Asn-110 are essential for secretion of human diamine oxidase. 29187599 6 75 theme human 991:995 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 3 76 dep carry 431:435 arg1 whereas 459:465 whereas 459:465 whereas 459:465 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 12 77 theme functional 1711:1720 arg1 results 1722:1728 These functional results 1705:1728 These functional results 1705:1728 These functional results are reflected in the high evolutionary conservation of all four glycosylation sites. 29187599 6 78 theme amine 977:981 arg1 protein-1 1015:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 947:1023 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 3 79 dep sites 367:371 arg1 sites 367:371 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 347:401 the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) 347:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 79 dep sites 367:371 arg1 Asn-538 382:388 Asn-538 382:388 Asn-538 382:388 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 79 dep sites 367:371 arg1 Asn-745 395:401 Asn-745 395:401 Asn-745 395:401 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 79 dep sites 367:371 arg1 Asn-168 373:379 Asn-168 373:379 Asn-168 373:379 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 cells 605:609 Caco-2 cells 598:609 Caco-2 cells 598:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 hDAO 569:572 native hDAO 562:572 native hDAO from amniotic fluid and Caco-2 cells 562:609 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 rhDAO 642:646 rhDAO 642:646 rhDAO produced in two different HEK293 cell lines 642:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 DAO 612:614 DAO 612:614 DAO 612:614 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 fluid 588:592 amniotic fluid 579:592 amniotic fluid 579:592 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 4 80 from Asn-110 551:557 arg1 kidneys 629:635 porcine kidneys 621:635 porcine kidneys 621:635 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 8 81 from retention 1327:1335 arg1 reticulum 1356:1364 the endoplasmic reticulum 1340:1364 the endoplasmic reticulum 1340:1364 Asn-110 replacement with Gln completely abrogated rhDAO secretion and caused retention in the endoplasmic reticulum. 29187599 3 82 theme recombinant 406:416 arg1 rhDAO 424:428 rhDAO 424:428 rhDAO 424:428 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 82 theme recombinant 406:416 arg1 hDAO 418:421 recombinant hDAO 406:421 recombinant hDAO (rhDAO) 406:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 7 83 theme hydrophobic 1191:1201 arg1 cleft 1203:1207 a highly conserved hydrophobic cleft 1172:1207 a highly conserved hydrophobic cleft interacting with the core trisaccharide 1172:1247 X-ray structures revealed that the residues adjacent to Asn-110 and Asn-137 form a highly conserved hydrophobic cleft interacting with the core trisaccharide. 29187599 3 84 from sites 367:371 arg1 rhDAO 424:428 rhDAO 424:428 rhDAO 424:428 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 3 84 from sites 367:371 arg1 hDAO 418:421 recombinant hDAO 406:421 recombinant hDAO (rhDAO) 406:429 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 10 85 theme Asn-538/745 1468:1478 arg1 substitutions 1514:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions reduced rhDAO secretion by 85 and 94%. 29187599 4 86 theme cell 681:684 arg1 lines 686:690 two different HEK293 cell lines 660:690 two different HEK293 cell lines 660:690 Here, we show that Asn-110 in native hDAO from amniotic fluid and Caco-2 cells, DAO from porcine kidneys, and rhDAO produced in two different HEK293 cell lines is also consistently occupied by oligomannosidic glycans. 29187599 13 87 theme gastrointestinal 1849:1864 arg1 tract 1866:1870 the gastrointestinal tract 1845:1870 the gastrointestinal tract 1845:1870 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 5 88 theme cell 916:919 arg1 variations 935:944 some tissue- and cell line-specific variations 899:944 variations 935:944 Glycans at Asn-168 were predominantly sialylated with bi- to tetra-antennary branches, and Asn-538 and Asn-745 had similar complex-type glycans with some tissue- and cell line-specific variations. 29187599 6 89 gly glycosylation 1065:1077 arg1 Asn-137 1082:1088 Asn-137 1082:1088 Asn-137 1082:1088 The related copper-containing amine oxidase human vascular adhesion protein-1 also exclusively displayed high-mannose glycosylation at Asn-137. 29187599 10 90 theme triple 1507:1512 arg1 substitutions 1514:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions 1468:1526 Asn-538/745 double and Asn-168/538/745 triple substitutions reduced rhDAO secretion by 85 and 94%. 29187599 9 91 theme Asn-745 1402:1408 arg1 Mutations 1367:1375 Mutations 1367:1375 Mutations of Asn-168, Asn-538, and Asn-745 1367:1408 Mutations of Asn-168, Asn-538, and Asn-745 reduced rhDAO secretion by 13, 71, and 32%, respectively. 29187599 9 92 theme rhDAO 1418:1422 arg1 secretion 1424:1432 rhDAO secretion 1418:1432 rhDAO secretion 1418:1432 Mutations of Asn-168, Asn-538, and Asn-745 reduced rhDAO secretion by 13, 71, and 32%, respectively. 29187599 13 93 from placenta 1885:1892 arg1 abundant 1828:1835 abundant 1828:1835 abundant 1828:1835 Human DAO is abundant only in the gastrointestinal tract, kidney, and placenta, and glycosylation seems essential for reaching high enzyme expression levels in these tissues. 29187599 11 94 theme Asn-745 1628:1634 arg1 glycosylations 1636:1649 Asn-538 and Asn-745 glycosylations 1616:1649 Asn-538 and Asn-745 glycosylations 1616:1649 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 3 95 theme complex-type 437:448 arg1 glycans 450:456 complex-type glycans 437:456 complex-type glycans 437:456 We recently reported that the N-glycosylation sites Asn-168, Asn-538, and Asn-745 in recombinant hDAO (rhDAO) carry complex-type glycans, whereas Asn-110 carries only mammalian-atypical oligomannosidic glycans. 29187599 11 96 theme Asn-538 1616:1622 arg1 glycosylations 1636:1649 Asn-538 and Asn-745 glycosylations 1616:1649 Asn-538 and Asn-745 glycosylations 1616:1649 Because of their locations in the DAO structure, Asn-538 and Asn-745 glycosylations might be important for efficient DAO dimer formation. 29187599 2 97 contain has 225:227 arg2 sites 254:258 multiple N-glycosylation sites 229:258 multiple N-glycosylation sites 229:258 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 2 97 contain has 225:227 arg1 oxidase 174:180 Human diamine oxidase 160:180 Human diamine oxidase (hDAO) 160:187 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 29187599 2 97 contain has 225:227 arg1 hDAO 183:186 hDAO 183:186 hDAO 183:186 Human diamine oxidase (hDAO), required for histamine catabolism, has multiple N-glycosylation sites, but their roles, for example in DAO secretion, are unclear. 27938679 7 0 theme mobility 975:982 arg1 increase 947:954 The increase 943:954 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU 943:1016 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 10 1 theme EnvA129 1398:1404 arg1 mutant 1406:1411 the EnvA129 mutant 1394:1411 the EnvA129 mutant 1394:1411 Finally, anti-BLV serum inhibited syncytia formation even with the EnvA129 mutant. 27938679 2 2 from cattle 489:494 arg1 antibodies 460:469 antibodies 460:469 antibodies from BLV-infected cattle 460:494 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 2 from cattle 489:494 arg1 antibodies 438:447 monoclonal antibodies 427:447 monoclonal antibodies (MAbs) 427:454 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 2 from cattle 489:494 arg1 MAbs 450:453 MAbs 450:453 MAbs 450:453 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 5 3 gly glycosylated 744:755 arg1 sites 724:728 sites 724:728 sites 724:728 To determine which sites were actually glycosylated, we generated mutated SU forms, where each N-glycosylation site was changed to alanine (A). 27938679 1 4 theme envelope 135:142 arg1 Env 153:155 Env 153:155 Env 153:155 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 4 theme envelope 135:142 arg1 protein 144:150 The bovine leukaemia virus (BLV) envelope protein 102:150 The bovine leukaemia virus (BLV) envelope protein (Env) 102:156 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 4 theme envelope 135:142 arg1 precursor 190:198 a polyprotein precursor 176:198 a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins 176:295 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 9 5 theme SU 1295:1296 arg1 structure 1320:1328 amino-terminal SU glycan conformational structure 1280:1328 amino-terminal SU glycan conformational structure 1280:1328 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 0 6 theme viral 88:92 arg1 escape 94:99 viral escape 88:99 viral escape 88:99 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 5 7 theme SU 779:780 arg1 forms 782:786 mutated SU forms 771:786 mutated SU forms 771:786 To determine which sites were actually glycosylated, we generated mutated SU forms, where each N-glycosylation site was changed to alanine (A). 27938679 9 8 theme amino-terminal 1280:1293 arg1 structure 1320:1328 amino-terminal SU glycan conformational structure 1280:1328 amino-terminal SU glycan conformational structure 1280:1328 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 9 9 theme glycan 1298:1303 arg1 structure 1320:1328 amino-terminal SU glycan conformational structure 1280:1328 amino-terminal SU glycan conformational structure 1280:1328 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 7 10 gly glycosylated 1078:1089 arg1 N256 1054:1057 N256 1054:1057 N256 1054:1057 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 10 gly glycosylated 1078:1089 arg1 residues 1045:1052 the asparagine residues N256 and N271 1030:1066 the asparagine residues N256 and N271 1030:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 10 gly glycosylated 1078:1089 arg1 N271 1063:1066 N271 1063:1066 N271 1063:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 11 theme electrophoretic 959:973 arg1 mobility 975:982 electrophoretic mobility 959:982 electrophoretic mobility of EnvA256 and EnvA271 derived SU 959:1016 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 11 12 theme latter 1418:1423 arg1 higher 1440:1445 higher 1440:1445 higher 1440:1445 The latter inhibition was higher than Env, suggesting that the oligosaccharides could be also involved in the glycan shield for viral escape. 27938679 11 12 theme latter 1418:1423 arg1 inhibition 1425:1434 The latter inhibition 1414:1434 The latter inhibition 1414:1434 The latter inhibition was higher than Env, suggesting that the oligosaccharides could be also involved in the glycan shield for viral escape. 27938679 8 13 theme antigenic 1157:1165 arg1 conformation 1167:1178 the antigenic conformation 1153:1178 the antigenic conformation of SU 1153:1184 ELISA revealed that only the N129 oligosaccharide determined the antigenic conformation of SU. 27938679 7 14 theme asparagine 1034:1043 arg1 N256 1054:1057 N256 1054:1057 N256 1054:1057 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 14 theme asparagine 1034:1043 arg1 residues 1045:1052 the asparagine residues N256 and N271 1030:1066 the asparagine residues N256 and N271 1030:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 14 theme asparagine 1034:1043 arg1 N271 1063:1066 N271 1063:1066 N271 1063:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 10 15 theme syncytia 1365:1372 arg1 formation 1374:1382 syncytia formation 1365:1382 syncytia formation 1365:1382 Finally, anti-BLV serum inhibited syncytia formation even with the EnvA129 mutant. 27938679 6 16 theme Env 930:932 arg1 mutants 934:940 Env mutants 930:940 Env mutants 930:940 Subsequently, these N to A mutations were inserted into the env gene to generate Env mutants. 27938679 4 17 dep N203 594:597 arg1 The 584:586 The 584:586 The 584:586 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 2 18 theme conformational 339:352 arg1 F 363:363 F 363:363 F 363:363 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 18 theme conformational 339:352 arg1 H 372:372 H 372:372 H 372:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 18 theme conformational 339:352 arg1 epitopes 354:361 conformational epitopes F, G and H 339:372 conformational epitopes F, G and H 339:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 18 theme conformational 339:352 arg1 G 366:366 G 366:366 G 366:366 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 19 theme monoclonal 427:436 arg1 antibodies 438:447 monoclonal antibodies 427:447 monoclonal antibodies (MAbs) 427:454 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 19 theme monoclonal 427:436 arg1 MAbs 450:453 MAbs 450:453 MAbs 450:453 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 11 20 theme viral 1542:1546 arg1 escape 1548:1553 viral escape 1542:1553 viral escape 1542:1553 The latter inhibition was higher than Env, suggesting that the oligosaccharides could be also involved in the glycan shield for viral escape. 27938679 3 21 gly N-glycosylation 561:575 arg2 residues 534:541 eight asparagine (N) residues 513:541 eight asparagine (N) residues that are putative N-glycosylation sites 513:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 21 gly N-glycosylation 561:575 arg2 sites 577:581 putative N-glycosylation sites 552:581 putative N-glycosylation sites 552:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 4 22 theme essential 663:671 arg1 role 673:676 an essential role 660:676 an essential role 660:676 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 1 23 gly glycoproteins 283:295 arg1 glycoproteins 283:295 the mature surface (SU) and transmembrane (TM) glycoproteins 236:295 glycoproteins 283:295 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 23 gly glycoproteins 283:295 arg1 TM 279:280 TM 279:280 TM 279:280 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 4 24 dep in 685:686 arg1 vitro 688:692 vitro 688:692 vitro 688:692 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 0 25 theme N-glycosylation 7:21 arg1 site 23:26 Single N-glycosylation site 0:26 Single N-glycosylation site of bovine leukemia virus SU 0:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 0 25 theme N-glycosylation 7:21 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 6 26 dep mutations 876:884 arg1 to 871:872 to 871:872 to 871:872 Subsequently, these N to A mutations were inserted into the env gene to generate Env mutants. 27938679 9 27 theme syncytium 1191:1199 arg1 formation 1201:1209 The syncytium formation 1187:1209 The syncytium formation induced by EnvA129 1187:1228 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 4 28 theme carbohydrate 633:644 arg1 binding 646:652 carbohydrate binding 633:652 carbohydrate binding 633:652 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 2 29 theme SU 327:328 arg1 region 317:322 The amino-terminal region 298:322 The amino-terminal region of SU 298:328 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 0 30 theme Single 0:5 arg1 site 23:26 Single N-glycosylation site 0:26 Single N-glycosylation site of bovine leukemia virus SU 0:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 0 30 theme Single 0:5 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 7 31 theme derived 1007:1013 arg1 SU 1015:1016 EnvA256 and EnvA271 derived SU 987:1016 EnvA256 and EnvA271 derived SU 987:1016 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 10 32 theme anti-BLV 1340:1347 arg1 serum 1349:1353 anti-BLV serum 1340:1353 anti-BLV serum 1340:1353 Finally, anti-BLV serum inhibited syncytia formation even with the EnvA129 mutant. 27938679 4 33 dep appear 614:619 arg1 play 655:658 play 655:658 play an essential role in the in vitro infection 655:702 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 4 33 dep appear 614:619 arg1 involved 621:628 involved 621:628 appear involved in carbohydrate binding 614:652 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 6 34 theme N 869:869 arg1 mutations 876:884 these N to A mutations 863:884 these N to A mutations 863:884 Subsequently, these N to A mutations were inserted into the env gene to generate Env mutants. 27938679 7 35 theme SU 1015:1016 arg1 mobility 975:982 electrophoretic mobility 959:982 electrophoretic mobility of EnvA256 and EnvA271 derived SU 959:1016 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 1 36 theme mature 240:245 arg1 SU 256:257 the mature surface (SU) and transmembrane (TM) glycoproteins 236:295 SU 256:257 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 9 37 theme structure 1320:1328 arg1 independent 1265:1275 independent 1265:1275 independent 1265:1275 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 9 38 theme fusogenic 1242:1250 arg1 capacity 1252:1259 fusogenic capacity 1242:1259 fusogenic capacity 1242:1259 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 6 39 theme env 909:911 arg1 gene 913:916 the env gene 905:916 the env gene 905:916 Subsequently, these N to A mutations were inserted into the env gene to generate Env mutants. 27938679 1 40 theme surface 247:253 arg1 SU 256:257 the mature surface (SU) and transmembrane (TM) glycoproteins 236:295 SU 256:257 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 3 41 theme putative 552:559 arg1 sites 577:581 putative N-glycosylation sites 552:581 putative N-glycosylation sites 552:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 41 theme putative 552:559 arg1 residues 534:541 eight asparagine (N) residues 513:541 eight asparagine (N) residues that are putative N-glycosylation sites 513:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 9 42 theme conformational 1305:1318 arg1 structure 1320:1328 amino-terminal SU glycan conformational structure 1280:1328 amino-terminal SU glycan conformational structure 1280:1328 The syncytium formation induced by EnvA129 showed that fusogenic capacity was independent of amino-terminal SU glycan conformational structure. 27938679 8 43 theme SU 1183:1184 arg1 conformation 1167:1178 the antigenic conformation 1153:1178 the antigenic conformation of SU 1153:1184 ELISA revealed that only the N129 oligosaccharide determined the antigenic conformation of SU. 27938679 0 44 theme leukemia 38:45 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 3 45 theme N-glycosylation 561:575 arg1 sites 577:581 putative N-glycosylation sites 552:581 putative N-glycosylation sites 552:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 45 theme N-glycosylation 561:575 arg1 residues 534:541 eight asparagine (N) residues 513:541 eight asparagine (N) residues that are putative N-glycosylation sites 513:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 2 46 gly glycosylated 391:402 arg1 SU 404:405 a glycosylated SU 389:405 a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle 389:494 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 0 47 theme bovine 31:36 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 2 48 theme glycosylated 391:402 arg1 SU 404:405 a glycosylated SU 389:405 a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle 389:494 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 49 contain contains 330:337 arg2 G 366:366 G 366:366 G 366:366 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 49 contain contains 330:337 arg2 F 363:363 F 363:363 F 363:363 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 49 contain contains 330:337 arg2 epitopes 354:361 conformational epitopes F, G and H 339:372 conformational epitopes F, G and H 339:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 49 contain contains 330:337 arg2 H 372:372 H 372:372 H 372:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 49 contain contains 330:337 arg1 region 317:322 The amino-terminal region 298:322 The amino-terminal region of SU 298:328 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 0 50 gly N-glycosylation 7:21 arg2 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 0 50 gly N-glycosylation 7:21 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 0 50 gly N-glycosylation 7:21 arg2 site 23:26 Single N-glycosylation site 0:26 Single N-glycosylation site of bovine leukemia virus SU 0:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 5 51 gly N-glycosylation 800:814 arg2 site 816:819 each N-glycosylation site 795:819 each N-glycosylation site 795:819 To determine which sites were actually glycosylated, we generated mutated SU forms, where each N-glycosylation site was changed to alanine (A). 27938679 8 52 theme N129 1121:1124 arg1 oligosaccharide 1126:1140 only the N129 oligosaccharide 1112:1140 only the N129 oligosaccharide 1112:1140 ELISA revealed that only the N129 oligosaccharide determined the antigenic conformation of SU. 27938679 0 53 theme SU 53:54 arg1 site 23:26 Single N-glycosylation site 0:26 Single N-glycosylation site of bovine leukemia virus SU 0:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 0 53 theme SU 53:54 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 11 54 dep glycan 1524:1529 arg1 shield 1531:1536 shield 1531:1536 shield for viral escape 1531:1553 The latter inhibition was higher than Env, suggesting that the oligosaccharides could be also involved in the glycan shield for viral escape. 27938679 0 55 theme virus 47:51 arg1 SU 53:54 bovine leukemia virus SU 31:54 bovine leukemia virus SU 31:54 Single N-glycosylation site of bovine leukemia virus SU is involved in conformation and viral escape. 27938679 7 56 dep residues 1045:1052 arg1 N256 1054:1057 N256 1054:1057 N256 1054:1057 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 56 dep residues 1045:1052 arg1 residues 1045:1052 the asparagine residues N256 and N271 1030:1066 the asparagine residues N256 and N271 1030:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 56 dep residues 1045:1052 arg1 N271 1063:1066 N271 1063:1066 N271 1063:1066 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 7 57 link derived 1007:1013 arg1 SU 1015:1016 EnvA256 and EnvA271 derived SU 987:1016 EnvA256 and EnvA271 derived SU 987:1016 The increase of electrophoretic mobility of EnvA256 and EnvA271 derived SU showed that the asparagine residues N256 and N271 were also glycosylated. 27938679 1 58 theme bovine 106:111 arg1 BLV 130:132 BLV 130:132 BLV 130:132 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 58 theme bovine 106:111 arg1 virus 123:127 bovine leukaemia virus 106:127 The bovine leukaemia virus (BLV) envelope protein (Env) 102:156 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 59 theme transmembrane 264:276 arg1 glycoproteins 283:295 the mature surface (SU) and transmembrane (TM) glycoproteins 236:295 glycoproteins 283:295 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 59 theme transmembrane 264:276 arg1 TM 279:280 TM 279:280 TM 279:280 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 3 60 theme asparagine 519:528 arg1 sites 577:581 putative N-glycosylation sites 552:581 putative N-glycosylation sites 552:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 60 theme asparagine 519:528 arg1 residues 534:541 eight asparagine (N) residues 513:541 eight asparagine (N) residues that are putative N-glycosylation sites 513:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 2 61 dep epitopes 354:361 arg1 F 363:363 F 363:363 F 363:363 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 61 dep epitopes 354:361 arg1 H 372:372 H 372:372 H 372:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 61 dep epitopes 354:361 arg1 epitopes 354:361 conformational epitopes F, G and H 339:372 conformational epitopes F, G and H 339:372 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 2 61 dep epitopes 354:361 arg1 G 366:366 G 366:366 G 366:366 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 1 62 theme leukaemia 113:121 arg1 BLV 130:132 BLV 130:132 BLV 130:132 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 62 theme leukaemia 113:121 arg1 virus 123:127 bovine leukaemia virus 106:127 The bovine leukaemia virus (BLV) envelope protein (Env) 102:156 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 63 theme polyprotein 178:188 arg1 protein 144:150 The bovine leukaemia virus (BLV) envelope protein 102:150 The bovine leukaemia virus (BLV) envelope protein (Env) 102:156 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 63 theme polyprotein 178:188 arg1 gp72 201:204 gp72 201:204 gp72 201:204 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 63 theme polyprotein 178:188 arg1 precursor 190:198 a polyprotein precursor 176:198 a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins 176:295 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 2 64 theme BLV-infected 476:487 arg1 cattle 489:494 BLV-infected cattle 476:494 BLV-infected cattle 476:494 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 27938679 1 65 theme virus 123:127 arg1 Env 153:155 Env 153:155 Env 153:155 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 65 theme virus 123:127 arg1 protein 144:150 The bovine leukaemia virus (BLV) envelope protein 102:150 The bovine leukaemia virus (BLV) envelope protein (Env) 102:156 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 1 65 theme virus 123:127 arg1 precursor 190:198 a polyprotein precursor 176:198 a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins 176:295 The bovine leukaemia virus (BLV) envelope protein (Env) is synthesized as a polyprotein precursor (gp72) proteolytically cleaved into the mature surface (SU) and transmembrane (TM) glycoproteins. 27938679 5 66 theme N-glycosylation 800:814 arg1 site 816:819 each N-glycosylation site 795:819 each N-glycosylation site 795:819 To determine which sites were actually glycosylated, we generated mutated SU forms, where each N-glycosylation site was changed to alanine (A). 27938679 5 67 theme mutated 771:777 arg1 forms 782:786 mutated SU forms 771:786 mutated SU forms 771:786 To determine which sites were actually glycosylated, we generated mutated SU forms, where each N-glycosylation site was changed to alanine (A). 27938679 6 68 theme A 874:874 arg1 mutations 876:884 these N to A mutations 863:884 these N to A mutations 863:884 Subsequently, these N to A mutations were inserted into the env gene to generate Env mutants. 27938679 3 69 contain contains 504:511 arg2 residues 534:541 eight asparagine (N) residues 513:541 eight asparagine (N) residues that are putative N-glycosylation sites 513:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 69 contain contains 504:511 arg1 SU 501:502 The SU 497:502 The SU 497:502 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 3 69 contain contains 504:511 arg2 sites 577:581 putative N-glycosylation sites 552:581 putative N-glycosylation sites 552:581 The SU contains eight asparagine (N) residues that are putative N-glycosylation sites. 27938679 4 70 theme in 685:686 arg1 infection 694:702 the in vitro infection 681:702 the in vitro infection 681:702 The N129, N203, N230 and N251 appear involved in carbohydrate binding, play an essential role in the in vitro infection. 27938679 2 71 theme amino-terminal 302:315 arg1 region 317:322 The amino-terminal region 298:322 The amino-terminal region of SU 298:328 The amino-terminal region of SU contains conformational epitopes F, G and H, which require a glycosylated SU to be recognized by monoclonal antibodies (MAbs) and antibodies from BLV-infected cattle. 24130173 6 0 theme comparable 1181:1190 arg1 activity 1172:1179 functional in vitro activity 1152:1179 functional in vitro activity comparable to that of its commercially available equine-derived counterpart 1152:1255 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 4 1 theme secreted 616:623 arg1 rBChE 643:647 rBChE 643:647 rBChE 643:647 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 4 1 theme secreted 616:623 arg1 BChE 637:640 secreted recombinant BChE 616:640 secreted recombinant BChE (rBChE) collected from the intercellular fluid 616:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 2 2 theme challenging 362:372 arg1 issue 374:378 a challenging issue 360:378 a challenging issue in respect to its recombinant expression 360:419 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 2 theme challenging 362:372 arg1 presence 273:280 presence 273:280 presence 273:280 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 2 theme challenging 362:372 arg1 necessity 297:305 functional necessity 286:305 functional necessity 286:305 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 4 3 theme BChE 637:640 arg1 profiling 603:611 Site-specific sugar profiling 583:611 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid 583:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 7 4 theme sialylated 1350:1359 arg1 profiles 1375:1382 designed sialylated glycosylation profiles 1341:1382 designed sialylated glycosylation profiles optimized for therapeutic efficacy 1341:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 8 5 theme present 1471:1477 arg1 carbohydrates 1457:1469 carbohydrates 1457:1469 carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) 1457:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 2 6 theme functional 286:295 arg1 issue 374:378 a challenging issue 360:378 a challenging issue in respect to its recombinant expression 360:419 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 6 theme functional 286:295 arg1 presence 273:280 presence 273:280 presence 273:280 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 6 theme functional 286:295 arg1 necessity 297:305 functional necessity 286:305 functional necessity 286:305 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 1 7 theme nerve 202:206 arg1 agents 208:213 nerve agents 202:213 nerve agents 202:213 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 5 8 theme branched 938:945 arg1 N-glycans 947:955 branched N-glycans 938:955 branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 938:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 8 theme branched 938:945 arg1 IV 1001:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 2 9 theme N-glycans 318:326 arg1 issue 374:378 a challenging issue 360:378 a challenging issue in respect to its recombinant expression 360:419 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 9 theme N-glycans 318:326 arg1 presence 273:280 presence 273:280 presence 273:280 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 9 theme N-glycans 318:326 arg1 necessity 297:305 functional necessity 286:305 functional necessity 286:305 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 5 10 theme glycosylation 902:914 arg1 enzyme 916:921 an additional glycosylation enzyme 888:921 an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 888:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 1 11 from use 219:221 arg1 treatment 249:257 pre- and post-exposure treatment 226:257 pre- and post-exposure treatment 226:257 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 8 12 theme efficient 1434:1442 arg1 synthesis 1444:1452 the efficient synthesis 1430:1452 the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) 1430:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 13 attach present 1471:1477 arg2 carbohydrates 1457:1469 carbohydrates 1457:1469 carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) 1457:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 13 attach present 1471:1477 arg1 amounts 1494:1500 minute amounts 1487:1500 minute amounts on the native protein (tri-sialylated N-glycans) 1487:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 3 14 theme protein 562:568 arg1 sialylation 570:580 in planta protein sialylation 552:580 in planta protein sialylation 552:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 8 15 theme new 1623:1625 arg1 functions 1639:1647 new therapeutic functions 1623:1647 new therapeutic functions 1623:1647 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 7 16 theme designed 1341:1348 arg1 profiles 1375:1382 designed sialylated glycosylation profiles 1341:1382 designed sialylated glycosylation profiles optimized for therapeutic efficacy 1341:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 4 17 theme N-glycans 738:746 arg1 presence 702:709 the presence 698:709 the presence 698:709 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 5 18 theme β1,4-N-acetylglucosaminyl-transferase 963:999 arg1 N-glycans 947:955 branched N-glycans 938:955 branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 938:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 18 theme β1,4-N-acetylglucosaminyl-transferase 963:999 arg1 IV 1001:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 4 19 theme plasma-derived 780:793 arg1 orthologue 795:804 the plasma-derived orthologue 776:804 the plasma-derived orthologue 776:804 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 7 20 theme plants 1299:1304 arg1 ability 1288:1294 the ability 1284:1294 the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy 1284:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 8 21 from present 1471:1477 arg1 amounts 1494:1500 minute amounts 1487:1500 minute amounts on the native protein (tri-sialylated N-glycans) 1487:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 6 22 theme in 1163:1164 arg1 activity 1172:1179 functional in vitro activity 1152:1179 functional in vitro activity comparable to that of its commercially available equine-derived counterpart 1152:1255 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 3 23 theme plant 478:482 arg1 benthamiana 494:504 the model plant Nicotiana benthamiana 468:504 the model plant Nicotiana benthamiana 468:504 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 0 24 theme human 14:18 arg1 butyrylcholinesterase 20:40 human butyrylcholinesterase 14:40 human butyrylcholinesterase 14:40 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 5 25 theme rBChE 1032:1036 arg1 production 1018:1027 the production 1014:1027 the production of rBChE decorated with tri-sialylated structures (up to 70%) 1014:1089 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 6 26 theme plant-derived 1122:1134 arg1 rBChE 1136:1140 non-sialylated plant-derived rBChE 1107:1140 non-sialylated plant-derived rBChE 1107:1140 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 4 27 theme sugar 597:601 arg1 profiling 603:611 Site-specific sugar profiling 583:611 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid 583:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 8 28 theme tri-sialylated 1525:1538 arg1 N-glycans 1540:1548 tri-sialylated N-glycans 1525:1548 tri-sialylated N-glycans 1525:1548 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 28 theme tri-sialylated 1525:1538 arg1 protein 1516:1522 the native protein 1505:1522 the native protein (tri-sialylated N-glycans) 1505:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 5 29 theme sialylation 833:843 arg1 content 845:851 that sialylation content 828:851 that sialylation content of rBChE 828:860 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 7 30 gly sialylated 1350:1359 arg1 profiles 1375:1382 designed sialylated glycosylation profiles 1341:1382 designed sialylated glycosylation profiles optimized for therapeutic efficacy 1341:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 0 31 link plasma-derived 98:111 arg1 orthologue 113:122 the plasma-derived orthologue 94:122 the plasma-derived orthologue 94:122 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 0 32 theme engineered 50:59 arg1 profile 75:81 an engineered glycosylation profile 47:81 an engineered glycosylation profile 47:81 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 0 33 with Expression 0:9 arg1 profile 75:81 an engineered glycosylation profile 47:81 an engineered glycosylation profile 47:81 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 8 34 with product 1583:1589 arg1 functions 1639:1647 new therapeutic functions 1623:1647 new therapeutic functions 1623:1647 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 34 with product 1583:1589 arg1 efficacies 1605:1614 superior efficacies 1596:1614 superior efficacies 1596:1614 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 1 35 theme post-exposure 235:247 arg1 treatment 249:257 pre- and post-exposure treatment 226:257 pre- and post-exposure treatment 226:257 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 3 36 theme planta 555:560 arg1 sialylation 570:580 in planta protein sialylation 552:580 in planta protein sialylation 552:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 7 37 theme therapeutic 1398:1408 arg1 efficacy 1410:1417 therapeutic efficacy 1398:1417 therapeutic efficacy 1398:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 6 38 dep available 1220:1228 arg1 equine-derived 1230:1243 equine-derived 1230:1243 equine-derived 1230:1243 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 3 39 theme necessary 538:546 arg1 genes 532:536 the genes 528:536 the genes necessary for in planta protein sialylation 528:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 4 40 link plasma-derived 780:793 arg1 orthologue 795:804 the plasma-derived orthologue 776:804 the plasma-derived orthologue 776:804 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 7 41 theme valuable 1318:1325 arg1 proteins 1327:1334 valuable proteins 1318:1334 valuable proteins 1318:1334 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 7 42 theme glycosylation 1361:1373 arg1 profiles 1375:1382 designed sialylated glycosylation profiles 1341:1382 designed sialylated glycosylation profiles optimized for therapeutic efficacy 1341:1417 These results demonstrate the ability of plants to generate valuable proteins with designed sialylated glycosylation profiles optimized for therapeutic efficacy. 24130173 2 43 theme complex 310:316 arg1 structures 345:354 sialylated structures 334:354 sialylated structures 334:354 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 43 theme complex 310:316 arg1 N-glycans 318:326 complex N-glycans 310:326 complex N-glycans (i.e. sialylated structures) 310:355 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 5 44 theme i.e. 958:961 arg1 N-glycans 947:955 branched N-glycans 938:955 branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 938:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 44 theme i.e. 958:961 arg1 IV 1001:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 i.e. β1,4-N-acetylglucosaminyl-transferase IV 958:1002 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 4 45 theme recombinant 625:635 arg1 rBChE 643:647 rBChE 643:647 rBChE 643:647 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 4 45 theme recombinant 625:635 arg1 BChE 637:640 secreted recombinant BChE 616:640 secreted recombinant BChE (rBChE) collected from the intercellular fluid 616:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 3 46 theme in 552:553 arg1 sialylation 570:580 in planta protein sialylation 552:580 in planta protein sialylation 552:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 5 47 theme enzyme 916:921 arg1 over-expression 869:883 the over-expression 865:883 the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 865:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 2 48 from issue 374:378 arg1 respect 383:389 respect 383:389 respect to its recombinant expression 383:419 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 6 49 link plant-derived 1122:1134 arg1 rBChE 1136:1140 non-sialylated plant-derived rBChE 1107:1140 non-sialylated plant-derived rBChE 1107:1140 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 2 50 theme sialylated 334:343 arg1 structures 345:354 sialylated structures 334:354 sialylated structures 334:354 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 50 theme sialylated 334:343 arg1 N-glycans 318:326 complex N-glycans 310:326 complex N-glycans (i.e. sialylated structures) 310:355 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 1 51 theme agents 208:213 arg1 bioscavenger 186:197 a candidate bioscavenger 174:197 a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment 174:257 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 2 52 dep presence 273:280 arg1 the 269:271 the 269:271 the 269:271 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 8 53 theme product 1583:1589 arg1 generation 1567:1576 the generation 1563:1576 the generation of a product with superior efficacies and/or new therapeutic functions 1563:1647 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 54 theme carbohydrates 1457:1469 arg1 synthesis 1444:1452 the efficient synthesis 1430:1452 the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) 1430:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 5 55 dep 70 1086:1087 arg1 to 1083:1084 to 1083:1084 to 1083:1084 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 56 theme additional 891:900 arg1 enzyme 916:921 an additional glycosylation enzyme 888:921 an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV) 888:1003 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 57 dep Attempts 807:814 arg1 allowed 1006:1012 allowed 1006:1012 allowed the production of rBChE decorated with tri-sialylated structures (up to 70%) 1006:1089 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 8 58 theme superior 1596:1603 arg1 efficacies 1605:1614 superior efficacies 1596:1614 superior efficacies 1596:1614 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 4 59 theme intercellular 669:681 arg1 fluid 683:687 the intercellular fluid 665:687 the intercellular fluid 665:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 6 60 dep in 1163:1164 arg1 vitro 1166:1170 vitro 1166:1170 vitro 1166:1170 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 4 61 gly di-sialylated 724:736 arg1 N-glycans 738:746 mono- and di-sialylated N-glycans 714:746 mono- and di-sialylated N-glycans 714:746 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 4 62 theme mono- 714:718 arg1 N-glycans 738:746 mono- and di-sialylated N-glycans 714:746 mono- and di-sialylated N-glycans 714:746 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 0 63 theme plasma-derived 98:111 arg1 orthologue 113:122 the plasma-derived orthologue 94:122 the plasma-derived orthologue 94:122 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 1 64 theme candidate 176:184 arg1 bioscavenger 186:197 a candidate bioscavenger 174:197 a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment 174:257 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 5 65 gly tri-sialylated 1053:1066 arg1 % 1088:1088 up to 70% 1080:1088 up to 70% 1080:1088 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 65 gly tri-sialylated 1053:1066 arg1 structures 1068:1077 tri-sialylated structures 1053:1077 tri-sialylated structures (up to 70%) 1053:1089 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 4 66 theme di-sialylated 724:736 arg1 N-glycans 738:746 mono- and di-sialylated N-glycans 714:746 mono- and di-sialylated N-glycans 714:746 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 5 67 theme tri-sialylated 1053:1066 arg1 % 1088:1088 up to 70% 1080:1088 up to 70% 1080:1088 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 5 67 theme tri-sialylated 1053:1066 arg1 structures 1068:1077 tri-sialylated structures 1053:1077 tri-sialylated structures (up to 70%) 1053:1089 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 3 68 contain carrying 519:526 arg1 vectors 511:517 vectors 511:517 vectors carrying the genes necessary for in planta protein sialylation 511:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 3 68 contain carrying 519:526 arg2 genes 532:536 the genes 528:536 the genes necessary for in planta protein sialylation 528:580 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 8 69 theme minute 1487:1492 arg1 amounts 1494:1500 minute amounts 1487:1500 minute amounts on the native protein (tri-sialylated N-glycans) 1487:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 2 70 theme recombinant 398:408 arg1 expression 410:419 its recombinant expression 394:419 its recombinant expression 394:419 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 5 71 theme rBChE 856:860 arg1 content 845:851 that sialylation content 828:851 that sialylation content of rBChE 828:860 Attempts to increase that sialylation content of rBChE by the over-expression of an additional glycosylation enzyme that generates branched N-glycans (i.e. β1,4-N-acetylglucosaminyl-transferase IV), allowed the production of rBChE decorated with tri-sialylated structures (up to 70%). 24130173 6 72 theme functional 1152:1161 arg1 activity 1172:1179 functional in vitro activity 1152:1179 functional in vitro activity comparable to that of its commercially available equine-derived counterpart 1152:1255 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 4 73 theme Site-specific 583:595 arg1 profiling 603:611 Site-specific sugar profiling 583:611 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid 583:687 Site-specific sugar profiling of secreted recombinant BChE (rBChE) collected from the intercellular fluid revealed the presence of mono- and di-sialylated N-glycans, which largely resembles to the plasma-derived orthologue. 24130173 2 74 dep structures 345:354 arg1 i.e. 329:332 i.e. 329:332 i.e. 329:332 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 3 75 theme BChE 455:458 arg1 cDNA 460:463 BChE cDNA 455:463 BChE cDNA 455:463 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 0 76 theme butyrylcholinesterase 20:40 arg1 Expression 0:9 Expression 0:9 Expression of human butyrylcholinesterase with an engineered glycosylation profile 0:81 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 8 77 dep new 1623:1625 arg1 therapeutic 1627:1637 therapeutic 1627:1637 therapeutic 1627:1637 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 1 78 theme Human 125:129 arg1 BChE 154:157 BChE 154:157 BChE 154:157 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 1 78 theme Human 125:129 arg1 butyrylcholinesterase 131:151 Human butyrylcholinesterase 125:151 Human butyrylcholinesterase (BChE) 125:158 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 8 79 theme native 1509:1514 arg1 N-glycans 1540:1548 tri-sialylated N-glycans 1525:1548 tri-sialylated N-glycans 1525:1548 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 79 theme native 1509:1514 arg1 protein 1516:1522 the native protein 1505:1522 the native protein (tri-sialylated N-glycans) 1505:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 3 80 theme Nicotiana 484:492 arg1 benthamiana 494:504 the model plant Nicotiana benthamiana 468:504 the model plant Nicotiana benthamiana 468:504 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 1 81 theme pre- 226:229 arg1 treatment 249:257 pre- and post-exposure treatment 226:257 pre- and post-exposure treatment 226:257 Human butyrylcholinesterase (BChE) is considered a candidate bioscavenger of nerve agents for use in pre- and post-exposure treatment. 24130173 6 82 theme non-sialylated 1107:1120 arg1 rBChE 1136:1140 non-sialylated plant-derived rBChE 1107:1140 non-sialylated plant-derived rBChE 1107:1140 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 6 83 gly non-sialylated 1107:1120 arg1 rBChE 1136:1140 non-sialylated plant-derived rBChE 1107:1140 non-sialylated plant-derived rBChE 1107:1140 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 3 84 theme model 472:476 arg1 benthamiana 494:504 the model plant Nicotiana benthamiana 468:504 the model plant Nicotiana benthamiana 468:504 Here we transiently co-expressed BChE cDNA in the model plant Nicotiana benthamiana with vectors carrying the genes necessary for in planta protein sialylation. 24130173 0 85 theme glycosylation 61:73 arg1 profile 75:81 an engineered glycosylation profile 47:81 an engineered glycosylation profile 47:81 Expression of human butyrylcholinesterase with an engineered glycosylation profile resembling the plasma-derived orthologue. 24130173 6 86 theme available 1220:1228 arg1 counterpart 1245:1255 its commercially available equine-derived counterpart 1203:1255 its commercially available equine-derived counterpart 1203:1255 Sialylated and non-sialylated plant-derived rBChE exhibited functional in vitro activity comparable to that of its commercially available equine-derived counterpart. 24130173 8 87 from amounts 1494:1500 arg1 N-glycans 1540:1548 tri-sialylated N-glycans 1525:1548 tri-sialylated N-glycans 1525:1548 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 87 from amounts 1494:1500 arg1 present 1471:1477 present 1471:1477 present 1471:1477 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 87 from amounts 1494:1500 arg1 protein 1516:1522 the native protein 1505:1522 the native protein (tri-sialylated N-glycans) 1505:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 88 gly tri-sialylated 1525:1538 arg1 N-glycans 1540:1548 tri-sialylated N-glycans 1525:1548 tri-sialylated N-glycans 1525:1548 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 8 88 gly tri-sialylated 1525:1538 arg1 protein 1516:1522 the native protein 1505:1522 the native protein (tri-sialylated N-glycans) 1505:1549 Moreover, the efficient synthesis of carbohydrates present only in minute amounts on the native protein (tri-sialylated N-glycans) facilitates the generation of a product with superior efficacies and/or new therapeutic functions. 24130173 2 89 gly sialylated 334:343 arg1 structures 345:354 sialylated structures 334:354 sialylated structures 334:354 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 24130173 2 89 gly sialylated 334:343 arg1 N-glycans 318:326 complex N-glycans 310:326 complex N-glycans (i.e. sialylated structures) 310:355 However, the presence and functional necessity of complex N-glycans (i.e. sialylated structures) is a challenging issue in respect to its recombinant expression. 25587188 11 0 theme somatic 1727:1733 arg1 hypermutation 1735:1747 somatic hypermutation 1727:1747 somatic hypermutation 1727:1747 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 2 1 theme exclusion 361:369 arg1 chromatography 371:384 size exclusion chromatography 356:384 size exclusion chromatography 356:384 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 13 2 theme immune 1931:1936 arg1 response 1938:1945 the citrulline-specific immune response 1907:1945 the citrulline-specific immune response in RA 1907:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 5 3 theme Binding 674:680 arg1 affinity 682:689 Binding affinity 674:689 Binding affinity to citrullinated antigens 674:715 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 11 4 theme N-glycosylation 1656:1670 arg1 sites 1672:1676 the N-glycosylation sites 1652:1676 the N-glycosylation sites in ACPA variable domains 1652:1701 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 3 5 theme gel 559:561 arg1 electrophoresis 563:577 gel electrophoresis 559:577 gel electrophoresis 559:577 ACPA-IgG and non-citrulline-specific IgG were affinity purified from serum, plasma and/or synovial fluid and analysed by gel electrophoresis. 25587188 2 6 theme RA 324:325 arg1 patients 327:334 ACPA-positive RA patients 310:334 ACPA-positive RA patients 310:334 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 5 7 theme plasmon 759:765 arg1 resonance 767:775 imaging surface plasmon resonance 743:775 imaging surface plasmon resonance 743:775 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 5 8 with ACPA 806:809 arg1 modifications 826:838 molecular modifications 816:838 molecular modifications 816:838 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 1 9 theme conventional 237:248 arg1 antibodies 251:260 'conventional' antibodies 236:260 'conventional' antibodies in rheumatoid arthritis (RA) 236:289 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 13 10 theme unique 1868:1873 arg1 feature 1896:1902 This unique and completely novel feature 1863:1902 This unique and completely novel feature of the citrulline-specific immune response in RA 1863:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 9 11 theme ACPA 1332:1335 arg1 avidity 1321:1327 binding avidity 1313:1327 binding avidity of ACPA to citrullinated antigens 1313:1361 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 5 12 theme imaging 743:749 arg1 resonance 767:775 imaging surface plasmon resonance 743:775 imaging surface plasmon resonance 743:775 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 9 13 theme citrullinated 1340:1352 arg1 antigens 1354:1361 citrullinated antigens 1340:1361 citrullinated antigens 1340:1361 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 7 14 theme disease-specific 1068:1083 arg1 autoantibodies 1085:1098 other disease-specific autoantibodies 1062:1098 other disease-specific autoantibodies 1062:1098 This feature also distinguished ACPA-IgG from antibodies against recall antigens or other disease-specific autoantibodies. 25587188 11 15 from sites 1672:1676 arg1 domains 1695:1701 ACPA variable domains 1681:1701 ACPA variable domains 1681:1701 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 12 16 theme B 1854:1854 arg1 cells 1856:1860 ACPA-producing B cells 1839:1860 ACPA-producing B cells 1839:1860 This finding also suggests that ACPA-hyperglycosylation confers a selective advantage to ACPA-producing B cells. 25587188 11 17 gly N-glycosylation 1656:1670 arg2 sites 1672:1676 the N-glycosylation sites 1652:1676 the N-glycosylation sites in ACPA variable domains 1652:1701 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 1 18 from antibodies 251:260 arg1 RA 287:288 RA 287:288 RA 287:288 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 1 18 from antibodies 251:260 arg1 arthritis 276:284 rheumatoid arthritis 265:284 rheumatoid arthritis (RA) 265:289 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 11 19 theme domains 1620:1626 arg1 germline-counterparts 1574:1594 the 'germline-counterparts' 1569:1595 the 'germline-counterparts' of identified variable domains 1569:1626 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 13 20 from response 1938:1945 arg1 RA 1950:1951 RA 1950:1951 RA 1950:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 7 21 theme recall 1043:1048 arg1 antigens 1050:1057 recall antigens 1043:1057 recall antigens 1043:1057 This feature also distinguished ACPA-IgG from antibodies against recall antigens or other disease-specific autoantibodies. 25587188 1 22 theme anti-citrullinated 186:203 arg1 ACPA 225:228 ACPA 225:228 ACPA 225:228 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 1 22 theme anti-citrullinated 186:203 arg1 antibodies 213:222 anti-citrullinated protein antibodies 186:222 anti-citrullinated protein antibodies (ACPA) 186:229 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 12 23 theme selective 1816:1824 arg1 advantage 1826:1834 a selective advantage 1814:1834 a selective advantage to ACPA-producing B cells 1814:1860 This finding also suggests that ACPA-hyperglycosylation confers a selective advantage to ACPA-producing B cells. 25587188 11 24 theme identified 1600:1609 arg1 domains 1620:1626 identified variable domains 1600:1626 identified variable domains 1600:1626 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 0 25 theme rheumatoid 100:109 arg1 arthritis 111:119 rheumatoid arthritis 100:119 rheumatoid arthritis 100:119 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 8 26 theme high 1137:1140 arg1 responsible 1207:1217 responsible 1207:1217 responsible 1207:1217 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 26 theme high 1137:1140 arg1 frequency 1142:1150 a high frequency 1135:1150 a high frequency of N-glycans in the (hyper)variable domains of ACPA 1135:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 13 27 theme B 2000:2000 arg1 response 2007:2014 the underlying B cell response 1985:2014 the underlying B cell response 1985:2014 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 3 28 theme non-citrulline-specific 451:473 arg1 IgG 475:477 non-citrulline-specific IgG 451:477 non-citrulline-specific IgG 451:477 ACPA-IgG and non-citrulline-specific IgG were affinity purified from serum, plasma and/or synovial fluid and analysed by gel electrophoresis. 25587188 11 29 gly glycosylation 1487:1499 arg2 sites 1511:1515 glycosylation consensus sites 1487:1515 glycosylation consensus sites in the protein sequence 1487:1539 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 13 30 theme citrulline-specific 1911:1929 arg1 response 1938:1945 the citrulline-specific immune response 1907:1945 the citrulline-specific immune response in RA 1907:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 0 31 theme citrullinated 74:86 arg1 antigens 88:95 citrullinated antigens 74:95 citrullinated antigens 74:95 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 13 32 theme response 2007:2014 arg1 understanding 1968:1980 our understanding 1964:1980 our understanding of the underlying B cell response 1964:2014 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 0 33 theme Extensive 0:8 arg1 glycosylation 10:22 Extensive glycosylation 0:22 Extensive glycosylation of ACPA-IgG variable domains 0:51 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 4 34 theme Electrophoresis 580:594 arg1 bands 596:600 Electrophoresis bands 580:600 Electrophoresis bands 580:600 Electrophoresis bands were excised, enzymatically digested and analysed by mass spectrometry. 25587188 6 35 theme molecular 925:933 arg1 weight 935:940 a 10-20 kDa higher molecular weight 906:940 a 10-20 kDa higher molecular weight 906:940 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 10 36 theme vast 1381:1384 arg1 majority 1386:1393 The vast majority 1377:1393 The vast majority of ACPA-IgG 1377:1405 CONCLUSIONS: The vast majority of ACPA-IgG harbour N-glycans in their variable domains. 25587188 0 37 theme variable 36:43 arg1 domains 45:51 ACPA-IgG variable domains 27:51 ACPA-IgG variable domains 27:51 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 6 38 theme 10-20 kDa 908:916 arg1 weight 935:940 a 10-20 kDa higher molecular weight 906:940 a 10-20 kDa higher molecular weight 906:940 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 0 39 gly glycosylation 10:22 arg1 domains 45:51 ACPA-IgG variable domains 27:51 ACPA-IgG variable domains 27:51 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 2 40 theme size 356:359 arg1 chromatography 371:384 size exclusion chromatography 356:384 size exclusion chromatography 356:384 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 6 41 theme donor 857:861 arg1 samples 863:869 all donor samples 853:869 all donor samples studied (n=24) 853:884 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 6 41 theme donor 857:861 arg1 n=24 880:883 n=24 880:883 n=24 880:883 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 11 42 theme ACPA 1681:1684 arg1 domains 1695:1701 ACPA variable domains 1681:1701 ACPA variable domains 1681:1701 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 9 43 with line 1244:1247 arg1 localisation 1260:1271 their localisation 1254:1271 their localisation 1254:1271 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 8 44 theme ACPA 1199:1202 arg1 ACPA 1199:1202 ACPA 1199:1202 ACPA 1199:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 44 theme ACPA 1199:1202 arg1 domains 1188:1194 the (hyper)variable domains 1168:1194 the (hyper)variable domains of ACPA 1168:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 13 45 theme response 1938:1945 arg1 feature 1896:1902 This unique and completely novel feature 1863:1902 This unique and completely novel feature of the citrulline-specific immune response in RA 1863:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 4 46 theme mass 655:658 arg1 spectrometry 660:671 mass spectrometry 655:671 mass spectrometry 655:671 Electrophoresis bands were excised, enzymatically digested and analysed by mass spectrometry. 25587188 11 47 theme N-linked 1455:1462 arg1 glycosylation 1464:1476 N-linked glycosylation 1455:1476 N-linked glycosylation 1455:1476 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 5 48 theme citrullinated 694:706 arg1 antigens 708:715 citrullinated antigens 694:715 citrullinated antigens 694:715 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 10 49 theme variable 1434:1441 arg1 domains 1443:1449 their variable domains 1428:1449 their variable domains 1428:1449 CONCLUSIONS: The vast majority of ACPA-IgG harbour N-glycans in their variable domains. 25587188 3 50 theme synovial 528:535 arg1 fluid 537:541 synovial fluid 528:541 synovial fluid 528:541 ACPA-IgG and non-citrulline-specific IgG were affinity purified from serum, plasma and/or synovial fluid and analysed by gel electrophoresis. 25587188 6 51 theme non-autoreactive 956:971 arg1 IgG 973:975 non-autoreactive IgG 956:975 non-autoreactive IgG 956:975 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 11 52 theme consensus 1501:1509 arg1 sites 1511:1515 glycosylation consensus sites 1487:1515 glycosylation consensus sites in the protein sequence 1487:1539 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 1 53 theme protein 205:211 arg1 ACPA 225:228 ACPA 225:228 ACPA 225:228 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 1 53 theme protein 205:211 arg1 antibodies 213:222 anti-citrullinated protein antibodies 186:222 anti-citrullinated protein antibodies (ACPA) 186:229 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 8 54 theme Structural 1101:1110 arg1 analysis 1112:1119 Structural analysis 1101:1119 Structural analysis 1101:1119 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 13 55 from RA 1950:1951 arg1 feature 1896:1902 This unique and completely novel feature 1863:1902 This unique and completely novel feature of the citrulline-specific immune response in RA 1863:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 2 56 theme patients 327:334 arg1 Serum 301:305 Serum 301:305 Serum of ACPA-positive RA patients 301:334 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 8 57 from frequency 1142:1150 arg1 ACPA 1199:1202 ACPA 1199:1202 ACPA 1199:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 57 from frequency 1142:1150 arg1 domains 1188:1194 the (hyper)variable domains 1168:1194 the (hyper)variable domains of ACPA 1168:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 6 58 dep studied 871:877 arg1 samples 863:869 all donor samples 853:869 all donor samples studied (n=24) 853:884 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 6 58 dep studied 871:877 arg1 n=24 880:883 n=24 880:883 n=24 880:883 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 13 59 theme novel 1890:1894 arg1 feature 1896:1902 This unique and completely novel feature 1863:1902 This unique and completely novel feature of the citrulline-specific immune response in RA 1863:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 5 60 theme surface 751:757 arg1 resonance 767:775 imaging surface plasmon resonance 743:775 imaging surface plasmon resonance 743:775 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 2 61 theme ACPA-positive 310:322 arg1 patients 327:334 ACPA-positive RA patients 310:334 ACPA-positive RA patients 310:334 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 11 62 from sites 1511:1515 arg1 sequence 1532:1539 the protein sequence 1520:1539 the protein sequence 1520:1539 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 5 63 theme molecular 816:824 arg1 modifications 826:838 molecular modifications 816:838 molecular modifications 816:838 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 1 64 theme molecular 152:160 arg1 features 162:169 the molecular features 148:169 the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA) 148:289 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 12 65 theme ACPA-producing 1839:1852 arg1 cells 1856:1860 ACPA-producing B cells 1839:1860 ACPA-producing B cells 1839:1860 This finding also suggests that ACPA-hyperglycosylation confers a selective advantage to ACPA-producing B cells. 25587188 11 66 theme variable 1611:1618 arg1 domains 1620:1626 identified variable domains 1600:1626 identified variable domains 1600:1626 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 5 67 theme monoclonal 795:804 arg1 ACPA 806:809 recombinant monoclonal ACPA 783:809 recombinant monoclonal ACPA with molecular modifications 783:838 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 7 68 theme other 1062:1066 arg1 autoantibodies 1085:1098 other disease-specific autoantibodies 1062:1098 other disease-specific autoantibodies 1062:1098 This feature also distinguished ACPA-IgG from antibodies against recall antigens or other disease-specific autoantibodies. 25587188 5 69 theme recombinant 783:793 arg1 ACPA 806:809 recombinant monoclonal ACPA 783:809 recombinant monoclonal ACPA with molecular modifications 783:838 Binding affinity to citrullinated antigens was measured by ELISA and imaging surface plasmon resonance using recombinant monoclonal ACPA with molecular modifications. 25587188 0 70 from binding 63:69 arg1 arthritis 111:119 rheumatoid arthritis 100:119 rheumatoid arthritis 100:119 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 13 71 theme underlying 1989:1998 arg1 response 2007:2014 the underlying B cell response 1985:2014 the underlying B cell response 1985:2014 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 9 72 located found 1295:1299 arg1 line 1244:1247 line 1244:1247 line with their localisation 1244:1271 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 9 72 located found 1295:1299 arg2 N-glycans 1280:1288 these N-glycans 1274:1288 these N-glycans 1274:1288 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 13 73 from feature 1896:1902 arg1 RA 1950:1951 RA 1950:1951 RA 1950:1951 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 8 74 theme N-glycans 1155:1163 arg1 responsible 1207:1217 responsible 1207:1217 responsible 1207:1217 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 74 theme N-glycans 1155:1163 arg1 frequency 1142:1150 a high frequency 1135:1150 a high frequency of N-glycans in the (hyper)variable domains of ACPA 1135:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 10 75 from N-glycans 1415:1423 arg1 domains 1443:1449 their variable domains 1428:1449 their variable domains 1428:1449 CONCLUSIONS: The vast majority of ACPA-IgG harbour N-glycans in their variable domains. 25587188 13 76 theme cell 2002:2005 arg1 response 2007:2014 the underlying B cell response 1985:2014 the underlying B cell response 1985:2014 This unique and completely novel feature of the citrulline-specific immune response in RA elucidates our understanding of the underlying B cell response. 25587188 11 77 link N-linked 1455:1462 arg1 glycosylation 1464:1476 N-linked glycosylation 1455:1476 N-linked glycosylation 1455:1476 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 10 78 theme ACPA-IgG 1398:1405 arg1 majority 1386:1393 The vast majority 1377:1393 The vast majority of ACPA-IgG 1377:1405 CONCLUSIONS: The vast majority of ACPA-IgG harbour N-glycans in their variable domains. 25587188 9 79 theme binding 1313:1319 arg1 avidity 1321:1327 binding avidity 1313:1327 binding avidity of ACPA to citrullinated antigens 1313:1361 In line with their localisation, these N-glycans were found to modulate binding avidity of ACPA to citrullinated antigens. 25587188 0 80 theme ACPA-IgG 27:34 arg1 domains 45:51 ACPA-IgG variable domains 27:51 ACPA-IgG variable domains 27:51 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 8 81 theme hyper 1173:1177 arg1 ACPA 1199:1202 ACPA 1199:1202 ACPA 1199:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 81 theme hyper 1173:1177 arg1 domains 1188:1194 the (hyper)variable domains 1168:1194 the (hyper)variable domains of ACPA 1168:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 0 82 theme domains 45:51 arg1 glycosylation 10:22 Extensive glycosylation 0:22 Extensive glycosylation of ACPA-IgG variable domains 0:51 Extensive glycosylation of ACPA-IgG variable domains modulates binding to citrullinated antigens in rheumatoid arthritis. 25587188 8 83 theme variable 1179:1186 arg1 ACPA 1199:1202 ACPA 1199:1202 ACPA 1199:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 8 83 theme variable 1179:1186 arg1 domains 1188:1194 the (hyper)variable domains 1168:1194 the (hyper)variable domains of ACPA 1168:1202 Structural analysis revealed that a high frequency of N-glycans in the (hyper)variable domains of ACPA is responsible for this observation. 25587188 6 84 dep 10-20 kDa 908:916 arg1 higher 918:923 higher 918:923 higher 918:923 RESULTS: In all donor samples studied (n=24), ACPA-IgG exhibited a 10-20 kDa higher molecular weight compared with non-autoreactive IgG. 25587188 11 85 theme variable 1686:1693 arg1 domains 1695:1701 ACPA variable domains 1681:1701 ACPA variable domains 1681:1701 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 2 86 theme ACPA-IgG 419:426 arg1 presence 407:414 the presence 403:414 the presence of ACPA-IgG by ELISA 403:435 METHODS: Serum of ACPA-positive RA patients was fractionated by size exclusion chromatography and analysed for the presence of ACPA-IgG by ELISA. 25587188 11 87 theme protein 1524:1530 arg1 sequence 1532:1539 the protein sequence 1520:1539 the protein sequence 1520:1539 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 3 88 dep purified 493:500 arg1 affinity 484:491 affinity 484:491 affinity 484:491 ACPA-IgG and non-citrulline-specific IgG were affinity purified from serum, plasma and/or synovial fluid and analysed by gel electrophoresis. 25587188 11 89 theme glycosylation 1487:1499 arg1 sites 1511:1515 glycosylation consensus sites 1487:1515 glycosylation consensus sites in the protein sequence 1487:1539 As N-linked glycosylation requires glycosylation consensus sites in the protein sequence and as these are lacking in the 'germline-counterparts' of identified variable domains, our data indicate that the N-glycosylation sites in ACPA variable domains have been introduced by somatic hypermutation. 25587188 1 90 theme rheumatoid 265:274 arg1 RA 287:288 RA 287:288 RA 287:288 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25587188 1 90 theme rheumatoid 265:274 arg1 arthritis 276:284 rheumatoid arthritis 265:284 rheumatoid arthritis (RA) 265:289 OBJECTIVES: To understand the molecular features distinguishing anti-citrullinated protein antibodies (ACPA) from 'conventional' antibodies in rheumatoid arthritis (RA). 25781883 1 0 theme physiological 267:279 arg1 function 281:288 a still elusive physiological function 251:288 a still elusive physiological function 251:288 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 10 1 theme CMG2 1325:1328 arg1 glycosylation 1330:1342 CMG2 glycosylation 1325:1342 CMG2 glycosylation 1325:1342 It thus appears that CMG2 glycosylation provides a buffer towards genetic variation by promoting folding of the protein in the ER lumen. 25781883 9 2 link N-linked 1179:1186 arg1 sugars 1188:1193 N-linked sugars 1179:1193 N-linked sugars 1179:1193 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 9 3 theme Fibromatosis 1281:1292 arg1 Syndrome 1294:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 6 4 theme mutants 818:824 arg1 Expression 788:797 Expression 788:797 Expression of N-glycosylation mutants 788:824 Expression of N-glycosylation mutants reveals that CMG2 is less vulnerable to sugar loss. 25781883 0 5 from dependence 13:22 arg1 N-glycosylation 27:41 N-glycosylation 27:41 N-glycosylation 27:41 Differential dependence on N-glycosylation of anthrax toxin receptors CMG2 and TEM8. 25781883 7 6 theme receptor 998:1005 arg1 function 1007:1014 receptor function 998:1014 receptor function 998:1014 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 7 theme type 1028:1031 arg1 protein 1033:1039 the wild type protein 1019:1039 the wild type protein expressed in tissue culture cells 1019:1073 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 4 8 attach presence 494:501 arg1 vWA 572:574 the vWA 568:574 the vWA 568:574 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 8 attach presence 494:501 arg1 domains 588:594 Ig-like domains 580:594 Ig-like domains 580:594 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 8 attach presence 494:501 arg1 domains 559:565 two extracellular domains 541:565 two extracellular domains 541:565 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 8 attach presence 494:501 arg1 each 529:532 each 529:532 each 529:532 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 8 attach presence 494:501 arg2 glycan 519:524 at least one glycan 506:524 at least one glycan 506:524 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 9 theme glycan 519:524 arg1 presence 494:501 The presence 490:501 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, 490:595 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 9 theme glycan 519:524 arg1 necessary 607:615 necessary 607:615 necessary 607:615 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 9 10 theme N-linked 1179:1186 arg1 sugars 1188:1193 N-linked sugars 1179:1193 N-linked sugars 1179:1193 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 5 11 theme ER 758:759 arg1 machinery 777:785 the ER quality control machinery 754:785 the ER quality control machinery 754:785 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 2 12 theme N-glycosylation 331:345 arg1 importance 317:326 the importance 313:326 the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins 313:422 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 7 13 theme wild 1023:1026 arg1 protein 1033:1039 the wild type protein 1019:1039 the wild type protein expressed in tissue culture cells 1019:1073 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 9 14 theme sugars 1188:1193 arg1 presence 1167:1174 the presence 1163:1174 the presence of N-linked sugars 1163:1193 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 0 15 theme anthrax 46:52 arg1 toxin 54:58 anthrax toxin 46:58 anthrax toxin 46:58 Differential dependence on N-glycosylation of anthrax toxin receptors CMG2 and TEM8. 25781883 7 16 theme culture 1061:1067 arg1 cells 1069:1073 tissue culture cells 1054:1073 tissue culture cells 1054:1073 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 17 theme tissue 1054:1059 arg1 cells 1069:1073 tissue culture cells 1054:1073 tissue culture cells 1054:1073 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 10 18 theme protein 1416:1422 arg1 folding 1401:1407 folding 1401:1407 folding of the protein 1401:1422 It thus appears that CMG2 glycosylation provides a buffer towards genetic variation by promoting folding of the protein in the ER lumen. 25781883 4 19 theme cell 650:653 arg1 surface 655:661 the cell surface 646:661 the cell surface 646:661 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 7 20 theme domains 938:944 arg1 domains 938:944 the extracellular domains 920:944 the extracellular domains 920:944 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 20 theme domains 938:944 arg1 one 913:915 one 913:915 one 913:915 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 1 21 theme type 137:140 arg1 proteins 153:160 two type I membrane proteins 133:160 two type I membrane proteins 133:160 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 21 theme type 137:140 arg1 ANTXR 85:89 ANTXR 1 and 2 85:97 ANTXR 85:89 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 21 theme type 137:140 arg1 2 97:97 2 97:97 2 97:97 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 0 22 theme Differential 0:11 arg1 dependence 13:22 Differential dependence 0:22 Differential dependence on N-glycosylation of anthrax toxin 0:58 Differential dependence on N-glycosylation of anthrax toxin receptors CMG2 and TEM8. 25781883 1 23 theme anthrax 217:223 arg1 toxin 225:229 anthrax toxin 217:229 anthrax toxin receptors 217:239 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 7 24 contain has 953:955 arg1 absence 882:888 The absence 878:888 The absence of N-linked glycans in one of the extracellular domains 878:944 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 24 contain has 953:955 arg2 impact 964:969 little impact 957:969 little impact 957:969 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 1 25 theme I 142:142 arg1 proteins 153:160 two type I membrane proteins 133:160 two type I membrane proteins 133:160 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 25 theme I 142:142 arg1 ANTXR 85:89 ANTXR 1 and 2 85:97 ANTXR 85:89 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 25 theme I 142:142 arg1 2 97:97 2 97:97 2 97:97 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 2 26 theme ligand 375:380 arg1 binding 382:388 ligand binding 375:388 ligand binding 375:388 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 1 27 theme toxin 225:229 arg1 receptors 231:239 anthrax toxin receptors 217:239 anthrax toxin receptors 217:239 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 7 28 theme little 957:962 arg1 impact 964:969 little impact 957:969 little impact 957:969 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 1 29 theme membrane 144:151 arg1 proteins 153:160 two type I membrane proteins 133:160 two type I membrane proteins 133:160 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 29 theme membrane 144:151 arg1 ANTXR 85:89 ANTXR 1 and 2 85:97 ANTXR 85:89 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 1 29 theme membrane 144:151 arg1 2 97:97 2 97:97 2 97:97 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 8 30 from patients 1147:1154 arg1 fibroblasts 1124:1134 primary fibroblasts 1116:1134 primary fibroblasts from human patients 1116:1154 N-glycans do, however, seem required in primary fibroblasts from human patients. 25781883 9 31 from mutations 1222:1230 arg1 cmg2 1235:1238 cmg2 1235:1238 cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome 1235:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 7 32 theme protein 1033:1039 arg1 trafficking 983:993 trafficking 983:993 trafficking 983:993 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 32 theme protein 1033:1039 arg1 function 1007:1014 receptor function 998:1014 receptor function 998:1014 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 32 theme protein 1033:1039 arg1 folding 974:980 folding 974:980 folding 974:980 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 4 33 theme extracellular 545:557 arg1 domains 559:565 two extracellular domains 541:565 two extracellular domains 541:565 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 33 theme extracellular 545:557 arg1 vWA 572:574 the vWA 568:574 the vWA 568:574 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 33 theme extracellular 545:557 arg1 domains 588:594 Ig-like domains 580:594 Ig-like domains 580:594 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 2 34 theme proteins 415:422 arg1 binding 382:388 ligand binding 375:388 ligand binding 375:388 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 2 34 theme proteins 415:422 arg1 folding 350:356 folding 350:356 folding 350:356 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 2 34 theme proteins 415:422 arg1 trafficking 359:369 trafficking 359:369 trafficking 359:369 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 10 35 theme genetic 1370:1376 arg1 variation 1378:1386 genetic variation 1370:1386 genetic variation 1370:1386 It thus appears that CMG2 glycosylation provides a buffer towards genetic variation by promoting folding of the protein in the ER lumen. 25781883 4 36 theme Ig-like 580:586 arg1 domains 559:565 two extracellular domains 541:565 two extracellular domains 541:565 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 36 theme Ig-like 580:586 arg1 domains 588:594 Ig-like domains 580:594 Ig-like domains 580:594 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 2 37 theme related 407:413 arg1 proteins 415:422 these closely related proteins 393:422 these closely related proteins 393:422 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 7 38 link N-linked 893:900 arg1 glycans 902:908 N-linked glycans 893:908 N-linked glycans 893:908 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 5 39 theme N-linked 686:693 arg1 glycans 695:701 any N-linked glycans 682:701 any N-linked glycans 682:701 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 4 40 from presence 494:501 arg1 vWA 572:574 the vWA 568:574 the vWA 568:574 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 40 from presence 494:501 arg1 domains 559:565 two extracellular domains 541:565 two extracellular domains 541:565 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 40 from presence 494:501 arg1 each 529:532 each 529:532 each 529:532 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 4 40 from presence 494:501 arg1 domains 588:594 Ig-like domains 580:594 Ig-like domains 580:594 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 2 41 from importance 317:326 arg1 binding 382:388 ligand binding 375:388 ligand binding 375:388 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 2 41 from importance 317:326 arg1 folding 350:356 folding 350:356 folding 350:356 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 2 41 from importance 317:326 arg1 trafficking 359:369 trafficking 359:369 trafficking 359:369 Here we have analyzed the importance of N-glycosylation on folding, trafficking and ligand binding of these closely related proteins. 25781883 5 42 theme glycans 695:701 arg1 absence 671:677 the absence 667:677 the absence of any N-linked glycans 667:701 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 7 43 theme extracellular 924:936 arg1 domains 938:944 the extracellular domains 920:944 the extracellular domains 920:944 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 6 44 theme sugar 866:870 arg1 loss 872:875 sugar loss 866:875 sugar loss 866:875 Expression of N-glycosylation mutants reveals that CMG2 is less vulnerable to sugar loss. 25781883 8 45 theme primary 1116:1122 arg1 fibroblasts 1124:1134 primary fibroblasts 1116:1134 primary fibroblasts from human patients 1116:1154 N-glycans do, however, seem required in primary fibroblasts from human patients. 25781883 4 46 theme efficient 621:629 arg1 trafficking 631:641 efficient trafficking 621:641 efficient trafficking to the cell surface 621:661 The presence of at least one glycan on each of its two extracellular domains, the vWA and Ig-like domains, is indeed necessary for efficient trafficking to the cell surface. 25781883 0 47 theme toxin 54:58 arg1 dependence 13:22 Differential dependence 0:22 Differential dependence on N-glycosylation of anthrax toxin 0:58 Differential dependence on N-glycosylation of anthrax toxin receptors CMG2 and TEM8. 25781883 3 48 contain has 443:445 arg2 dependence 459:468 a stringent dependence 447:468 a stringent dependence on N-glycosylation 447:487 We find that TEM8 has a stringent dependence on N-glycosylation. 25781883 3 48 contain has 443:445 arg1 TEM8 438:441 TEM8 438:441 TEM8 438:441 We find that TEM8 has a stringent dependence on N-glycosylation. 25781883 6 49 theme N-glycosylation 802:816 arg1 mutants 818:824 N-glycosylation mutants 802:824 N-glycosylation mutants 802:824 Expression of N-glycosylation mutants reveals that CMG2 is less vulnerable to sugar loss. 25781883 5 50 link N-linked 686:693 arg1 glycans 695:701 any N-linked glycans 682:701 any N-linked glycans 682:701 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 7 51 theme N-linked 893:900 arg1 glycans 902:908 N-linked glycans 893:908 N-linked glycans 893:908 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 52 theme glycans 902:908 arg1 absence 882:888 The absence 878:888 The absence of N-linked glycans in one of the extracellular domains 878:944 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 5 53 theme quality 761:767 arg1 machinery 777:785 the ER quality control machinery 754:785 the ER quality control machinery 754:785 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 9 54 theme rare 1252:1255 arg1 Syndrome 1294:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 9 55 theme genetic 1257:1263 arg1 Syndrome 1294:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 10 56 theme ER 1431:1432 arg1 lumen 1434:1438 the ER lumen 1427:1438 the ER lumen 1427:1438 It thus appears that CMG2 glycosylation provides a buffer towards genetic variation by promoting folding of the protein in the ER lumen. 25781883 8 57 theme human 1141:1145 arg1 patients 1147:1154 human patients 1141:1154 human patients 1141:1154 N-glycans do, however, seem required in primary fibroblasts from human patients. 25781883 5 58 theme control 769:775 arg1 machinery 777:785 the ER quality control machinery 754:785 the ER quality control machinery 754:785 In the absence of any N-linked glycans, TEM8 fails to fold correctly and is recognized by the ER quality control machinery. 25781883 7 59 from absence 882:888 arg1 domains 938:944 the extracellular domains 920:944 the extracellular domains 920:944 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 7 59 from absence 882:888 arg1 one 913:915 one 913:915 one 913:915 The absence of N-linked glycans in one of the extracellular domains indeed has little impact on folding, trafficking or receptor function of the wild type protein expressed in tissue culture cells. 25781883 9 60 theme disease 1265:1271 arg1 Syndrome 1294:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 25781883 1 61 theme elusive 259:265 arg1 function 281:288 a still elusive physiological function 251:288 a still elusive physiological function 251:288 ANTXR 1 and 2, also known as TEM8 and CMG2, are two type I membrane proteins, which have been extensively studied for their role as anthrax toxin receptors, but with a still elusive physiological function. 25781883 3 62 theme stringent 449:457 arg1 dependence 459:468 a stringent dependence 447:468 a stringent dependence on N-glycosylation 447:487 We find that TEM8 has a stringent dependence on N-glycosylation. 25781883 3 63 from dependence 459:468 arg1 N-glycosylation 473:487 N-glycosylation 473:487 N-glycosylation 473:487 We find that TEM8 has a stringent dependence on N-glycosylation. 25781883 9 64 theme Hyaline 1273:1279 arg1 Syndrome 1294:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 the rare genetic disease Hyaline Fibromatosis Syndrome 1248:1301 Here, the presence of N-linked sugars increases the tolerance to mutations in cmg2 causing the rare genetic disease Hyaline Fibromatosis Syndrome. 28797655 6 0 contain had 986:988 arg1 lectins 951:957 6 lectins 949:957 6 lectins (e.g., PHA-E, PSA and DSA) 949:984 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 6 0 contain had 986:988 arg2 results 1002:1008 contrasting results 990:1008 contrasting results 990:1008 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 10 1 dep prefer 1465:1470 arg1 attach 1481:1486 attach 1481:1486 prefer entially attach to each other 1465:1500 We conclude that the existing SAα2-3/6Gal sialoglycans make the offspring of the H9N2 virus prefer entially attach to each other, which decreases the virulence. 28797655 11 2 theme same 1703:1706 arg1 strain 1718:1723 the same influenza strain 1699:1723 the same influenza strain from different hosts 1699:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 11 3 dep evolution 1581:1589 arg1 the 1577:1579 the 1577:1579 the 1577:1579 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 12 4 theme viral 1838:1842 arg1 transmission 1844:1855 viral transmission 1838:1855 viral transmission 1838:1855 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 4 5 theme lectin 639:644 arg1 microarray 646:655 the lectin microarray 635:655 the lectin microarray 635:655 The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS. 28797655 0 6 theme N-glycan 0:7 arg1 profiles 9:16 N-glycan profiles 0:16 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 0:109 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 10 7 theme H9N2 1454:1457 arg1 virus 1459:1463 the H9N2 virus 1450:1463 the H9N2 virus 1450:1463 We conclude that the existing SAα2-3/6Gal sialoglycans make the offspring of the H9N2 virus prefer entially attach to each other, which decreases the virulence. 28797655 3 8 theme lung 507:510 arg1 MRC-5 530:534 MRC-5 530:534 MRC-5 530:534 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 8 theme lung 507:510 arg1 cells 523:527 human embryonic lung fibroblast cells 491:527 human embryonic lung fibroblast cells (MRC-5) 491:535 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 12 9 theme molecular 1811:1819 arg1 mechanism 1821:1829 the molecular mechanism 1807:1829 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 6 10 theme contrasting 990:1000 arg1 results 1002:1008 contrasting results 990:1008 contrasting results 990:1008 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 4 11 theme viral 562:566 arg1 profiles 577:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS. 28797655 3 12 theme human 491:495 arg1 MRC-5 530:534 MRC-5 530:534 MRC-5 530:534 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 12 theme human 491:495 arg1 cells 523:527 human embryonic lung fibroblast cells 491:527 human embryonic lung fibroblast cells (MRC-5) 491:535 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 5 13 theme conserved 747:755 arg1 sites 773:777 six and five highly conserved N-glycosylation sites 727:777 sites 773:777 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 13 theme conserved 747:755 arg1 result 684:689 a result 682:689 a result 682:689 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 12 14 theme novel 1774:1778 arg1 way 1780:1782 a novel way 1772:1782 a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1772:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 6 15 theme expression 854:863 arg1 levels 865:870 increased expression levels 844:870 increased expression levels of the glycan structures in the MRC-5 844:908 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 7 16 from system 1039:1044 arg1 glycans 1018:1024 Eleven glycans 1011:1024 Eleven glycans from the ECE system 1011:1044 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 7 16 from system 1039:1044 arg1 glycans 1053:1059 13 glycans 1050:1059 13 glycans from the MRC-5 system 1050:1081 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 12 17 theme vital 1943:1947 arg1 information 1949:1959 vital information 1943:1959 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 8 18 theme Fucα-1,6GlcNAc 1128:1141 arg1 structure 1156:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 3 19 theme cells 523:527 arg1 system 537:542 the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system 452:542 the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system 452:542 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 12 20 from glycosylation 1893:1905 arg1 hosts 1926:1930 avian and human hosts 1910:1930 avian and human hosts 1910:1930 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 8 21 theme core 1143:1146 arg1 structure 1156:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 9 22 theme MRC-5 1359:1363 arg1 system 1365:1370 the MRC-5 system 1355:1370 the MRC-5 system 1355:1370 The SAα2-3/6Gal structures were highly expressed and Fucα1-2Galβ1-4GlcNAc structures were only observed in the MRC-5 system. 28797655 11 23 gly glycosylation 1553:1565 arg2 sites 1567:1571 the glycosylation sites 1549:1571 the glycosylation sites for the evolution and role of IAVs 1549:1606 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 7 24 theme MRC-5 1070:1074 arg1 system 1076:1081 the MRC-5 system 1066:1081 the MRC-5 system 1066:1081 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 1 25 theme host 142:145 arg1 infectivity 174:184 infectivity 174:184 infectivity 174:184 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 25 theme host 142:145 arg1 virulence 160:168 virulence 160:168 virulence 160:168 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 25 theme host 142:145 arg1 specificity 147:157 the host specificity 138:157 the host specificity 138:157 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 11 26 from Alterations 1534:1544 arg1 sites 1567:1571 the glycosylation sites 1549:1571 the glycosylation sites for the evolution and role of IAVs 1549:1606 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 0 27 theme embryonic 78:86 arg1 cells 104:108 human embryonic lung fibroblast cells 72:108 human embryonic lung fibroblast cells 72:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 6 28 theme glycan 879:884 arg1 structures 886:895 the glycan structures 875:895 the glycan structures 875:895 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 8 29 theme pentaantennary 1185:1198 arg1 N-glycans 1200:1208 pentaantennary N-glycans 1185:1208 pentaantennary N-glycans 1185:1208 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 2 30 theme virus 345:349 arg1 HA 310:311 HA 310:311 HA 310:311 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 30 theme virus 345:349 arg1 neuraminidase 318:330 neuraminidase 318:330 neuraminidase (NA) 318:335 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 30 theme virus 345:349 arg1 hemagglutinin 295:307 the hemagglutinin 291:307 the hemagglutinin (HA) 291:312 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 30 theme virus 345:349 arg1 NA 333:334 NA 333:334 NA 333:334 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 0 31 theme fibroblast 93:102 arg1 cells 104:108 human embryonic lung fibroblast cells 72:108 human embryonic lung fibroblast cells 72:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 12 32 gly glycosylation 1893:1905 arg1 hosts 1926:1930 avian and human hosts 1910:1930 avian and human hosts 1910:1930 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 5 33 theme six 727:729 arg1 sites 773:777 six and five highly conserved N-glycosylation sites 727:777 sites 773:777 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 33 theme six 727:729 arg1 result 684:689 a result 682:689 a result 682:689 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 6 34 from levels 865:870 arg1 MRC-5 904:908 the MRC-5 900:908 the MRC-5 900:908 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 2 35 theme phylogenetic 371:382 arg1 analysis 384:391 phylogenetic analysis 371:391 phylogenetic analysis 371:391 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 5 36 theme five 735:738 arg1 sites 773:777 six and five highly conserved N-glycosylation sites 727:777 sites 773:777 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 36 theme five 735:738 arg1 result 684:689 a result 682:689 a result 682:689 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 12 37 theme viral 1887:1891 arg1 glycosylation 1893:1905 viral glycosylation 1887:1905 viral glycosylation in avian and human hosts 1887:1930 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 11 38 theme glycosylation 1553:1565 arg1 sites 1567:1571 the glycosylation sites 1549:1571 the glycosylation sites for the evolution and role of IAVs 1549:1606 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 12 39 theme human 2013:2017 arg1 viruses 2019:2025 other human viruses 2007:2025 other human viruses 2007:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 0 40 theme avian 26:30 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 6 41 theme ECE 928:930 arg1 system 932:937 the ECE system 924:937 the ECE system 924:937 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 0 42 theme chicken 55:61 arg1 eggs 63:66 chicken eggs 55:66 chicken eggs 55:66 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 5 43 theme viruses 710:716 arg1 NA 699:700 NA 699:700 NA 699:700 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 43 theme viruses 710:716 arg1 HA 692:693 HA 692:693 HA 692:693 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 3 44 theme H9N2 418:421 arg1 subtypes 423:430 the H9N2 subtypes 414:430 the H9N2 subtypes 414:430 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 11 45 theme exact 1671:1675 arg1 structures 1684:1693 the exact glycan structures 1667:1693 the exact glycan structures for the same influenza strain from different hosts 1667:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 4 46 theme glycomic 605:612 arg1 method 614:619 a glycomic method 603:619 a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS 603:676 The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS. 28797655 10 47 theme SAα2-3/6Gal 1403:1413 arg1 sialoglycans 1415:1426 the existing SAα2-3/6Gal sialoglycans 1390:1426 the existing SAα2-3/6Gal sialoglycans 1390:1426 We conclude that the existing SAα2-3/6Gal sialoglycans make the offspring of the H9N2 virus prefer entially attach to each other, which decreases the virulence. 28797655 6 48 contain had 840:842 arg1 lectins 802:808 Sixteen lectins 794:808 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) 794:838 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 6 48 contain had 840:842 arg2 levels 865:870 increased expression levels 844:870 increased expression levels of the glycan structures in the MRC-5 844:908 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 11 49 theme influenza 1708:1716 arg1 strain 1718:1723 the same influenza strain 1699:1723 the same influenza strain from different hosts 1699:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 12 50 theme transmission 1844:1855 arg1 mechanism 1821:1829 the molecular mechanism 1807:1829 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 12 50 theme transmission 1844:1855 arg1 information 1949:1959 vital information 1943:1959 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 2 51 gly N-glycosylation 266:280 arg2 sites 282:286 N-glycosylation sites 266:286 N-glycosylation sites 266:286 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 12 52 theme virulence 1861:1869 arg1 mechanism 1821:1829 the molecular mechanism 1807:1829 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 12 52 theme virulence 1861:1869 arg1 information 1949:1959 vital information 1943:1959 the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses 1807:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 7 53 from system 1076:1081 arg1 glycans 1018:1024 Eleven glycans 1011:1024 Eleven glycans from the ECE system 1011:1044 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 7 53 from system 1076:1081 arg1 glycans 1053:1059 13 glycans 1050:1059 13 glycans from the MRC-5 system 1050:1081 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 6 54 dep MAL-II 817:822 arg1 e.g. 811:814 e.g. 811:814 e.g. 811:814 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 9 55 theme SAα2-3/6Gal 1252:1262 arg1 structures 1264:1273 The SAα2-3/6Gal structures 1248:1273 The SAα2-3/6Gal structures 1248:1273 The SAα2-3/6Gal structures were highly expressed and Fucα1-2Galβ1-4GlcNAc structures were only observed in the MRC-5 system. 28797655 3 56 theme eggs 476:479 arg1 system 537:542 the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system 452:542 the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system 452:542 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 8 57 located observed 1220:1227 arg1 system 1240:1245 the ECE system 1232:1245 the ECE system 1232:1245 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 8 57 located observed 1220:1227 arg2 N-glycans 1200:1208 pentaantennary N-glycans 1185:1208 pentaantennary N-glycans 1185:1208 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 10 58 theme virus 1459:1463 arg1 offspring 1437:1445 the offspring 1433:1445 the offspring of the H9N2 virus 1433:1463 We conclude that the existing SAα2-3/6Gal sialoglycans make the offspring of the H9N2 virus prefer entially attach to each other, which decreases the virulence. 28797655 3 59 theme embryonic 497:505 arg1 MRC-5 530:534 MRC-5 530:534 MRC-5 530:534 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 59 theme embryonic 497:505 arg1 cells 523:527 human embryonic lung fibroblast cells 491:527 human embryonic lung fibroblast cells (MRC-5) 491:535 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 5 60 gly N-glycosylation 757:771 arg2 sites 773:777 six and five highly conserved N-glycosylation sites 727:777 sites 773:777 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 60 gly N-glycosylation 757:771 arg2 result 684:689 a result 682:689 a result 682:689 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 1 61 gly N-glycosylation 111:125 arg1 host 142:145 the host specificity 138:157 the host specificity 138:157 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 8 62 theme ECE 1236:1238 arg1 system 1240:1245 the ECE system 1232:1245 the ECE system 1232:1245 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 2 63 theme sites 282:286 arg1 evolution 253:261 evolution 253:261 evolution 253:261 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 63 theme sites 282:286 arg1 distribution 236:247 distribution 236:247 distribution 236:247 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 9 64 theme Fucα1-2Galβ1-4GlcNAc 1301:1320 arg1 structures 1322:1331 Fucα1-2Galβ1-4GlcNAc structures 1301:1331 Fucα1-2Galβ1-4GlcNAc structures 1301:1331 The SAα2-3/6Gal structures were highly expressed and Fucα1-2Galβ1-4GlcNAc structures were only observed in the MRC-5 system. 28797655 9 65 located observed 1343:1350 arg2 structures 1322:1331 Fucα1-2Galβ1-4GlcNAc structures 1301:1331 Fucα1-2Galβ1-4GlcNAc structures 1301:1331 The SAα2-3/6Gal structures were highly expressed and Fucα1-2Galβ1-4GlcNAc structures were only observed in the MRC-5 system. 28797655 9 65 located observed 1343:1350 arg1 system 1365:1370 the MRC-5 system 1355:1370 the MRC-5 system 1355:1370 The SAα2-3/6Gal structures were highly expressed and Fucα1-2Galβ1-4GlcNAc structures were only observed in the MRC-5 system. 28797655 4 66 theme proliferated 549:560 arg1 profiles 577:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS. 28797655 5 67 theme N-glycosylation 757:771 arg1 sites 773:777 six and five highly conserved N-glycosylation sites 727:777 sites 773:777 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 5 67 theme N-glycosylation 757:771 arg1 result 684:689 a result 682:689 a result 682:689 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 6 68 theme increased 844:852 arg1 levels 865:870 increased expression levels 844:870 increased expression levels of the glycan structures in the MRC-5 844:908 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 0 69 from eggs 63:66 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 69 from eggs 63:66 arg1 profiles 9:16 N-glycan profiles 0:16 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 0:109 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 4 70 theme N-glycan 568:575 arg1 profiles 577:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles 545:584 The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS. 28797655 3 71 theme fibroblast 512:521 arg1 MRC-5 530:534 MRC-5 530:534 MRC-5 530:534 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 71 theme fibroblast 512:521 arg1 cells 523:527 human embryonic lung fibroblast cells 491:527 human embryonic lung fibroblast cells (MRC-5) 491:535 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 0 72 from cells 104:108 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 72 from cells 104:108 arg1 profiles 9:16 N-glycan profiles 0:16 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 0:109 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 12 73 theme avian 1910:1914 arg1 hosts 1926:1930 avian and human hosts 1910:1930 avian and human hosts 1910:1930 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 2 74 from distribution 236:247 arg1 HA 310:311 HA 310:311 HA 310:311 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 74 from distribution 236:247 arg1 neuraminidase 318:330 neuraminidase 318:330 neuraminidase (NA) 318:335 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 74 from distribution 236:247 arg1 hemagglutinin 295:307 the hemagglutinin 291:307 the hemagglutinin (HA) 291:312 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 74 from distribution 236:247 arg1 NA 333:334 NA 333:334 NA 333:334 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 12 75 theme human 1920:1924 arg1 hosts 1926:1930 avian and human hosts 1910:1930 avian and human hosts 1910:1930 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 11 76 theme IAVs 1603:1606 arg1 role 1595:1598 role 1595:1598 role 1595:1598 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 11 76 theme IAVs 1603:1606 arg1 evolution 1581:1589 evolution 1581:1589 evolution 1581:1589 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 1 77 theme influenza 189:197 arg1 IAVs 210:213 IAVs 210:213 IAVs 210:213 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 77 theme influenza 189:197 arg1 viruses 201:207 influenza A viruses 189:207 influenza A viruses (IAVs) 189:214 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 2 78 from evolution 253:261 arg1 HA 310:311 HA 310:311 HA 310:311 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 78 from evolution 253:261 arg1 neuraminidase 318:330 neuraminidase 318:330 neuraminidase (NA) 318:335 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 78 from evolution 253:261 arg1 hemagglutinin 295:307 the hemagglutinin 291:307 the hemagglutinin (HA) 291:312 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 2 78 from evolution 253:261 arg1 NA 333:334 NA 333:334 NA 333:334 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 11 79 from hosts 1740:1744 arg1 strain 1718:1723 the same influenza strain 1699:1723 the same influenza strain from different hosts 1699:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 8 80 theme fucose 1148:1153 arg1 structure 1156:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 the Fucα-1,6GlcNAc(core fucose) structure 1124:1164 Our results showed that the Fucα-1,6GlcNAc(core fucose) structure was increased, and pentaantennary N-glycans were only observed in the ECE system. 28797655 0 81 theme human 72:76 arg1 cells 104:108 human embryonic lung fibroblast cells 72:108 human embryonic lung fibroblast cells 72:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 82 from profiles 9:16 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 82 from profiles 9:16 arg1 eggs 63:66 chicken eggs 55:66 chicken eggs 55:66 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 82 from profiles 9:16 arg1 cells 104:108 human embryonic lung fibroblast cells 72:108 human embryonic lung fibroblast cells 72:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 0 83 theme lung 88:91 arg1 cells 104:108 human embryonic lung fibroblast cells 72:108 human embryonic lung fibroblast cells 72:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 2 84 theme N-glycosylation 266:280 arg1 sites 282:286 N-glycosylation sites 266:286 N-glycosylation sites 266:286 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 6 85 theme structures 886:895 arg1 levels 865:870 increased expression levels 844:870 increased expression levels of the glycan structures in the MRC-5 844:908 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 3 86 theme subtypes 423:430 arg1 strain 404:409 one strain 400:409 one strain of the H9N2 subtypes 400:430 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 7 87 theme ECE 1035:1037 arg1 system 1039:1044 the ECE system 1031:1044 the ECE system 1031:1044 Eleven glycans from the ECE system and 13 glycans from the MRC-5 system were identified. 28797655 1 88 theme A 199:199 arg1 IAVs 210:213 IAVs 210:213 IAVs 210:213 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 88 theme A 199:199 arg1 viruses 201:207 influenza A viruses 189:207 influenza A viruses (IAVs) 189:214 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 2 89 theme H9N2 340:343 arg1 virus 345:349 H9N2 virus 340:349 H9N2 virus 340:349 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 11 90 theme different 1730:1738 arg1 hosts 1740:1744 different hosts 1730:1744 different hosts 1730:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 0 91 theme H9N2 21:24 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 3 92 theme embryonated 456:466 arg1 ECE 482:484 ECE 482:484 ECE 482:484 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 92 theme embryonated 456:466 arg1 eggs 476:479 embryonated chicken eggs 456:479 embryonated chicken eggs (ECE) 456:485 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 0 93 theme influenza 32:40 arg1 viruses 42:48 H9N2 avian influenza viruses 21:48 H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells 21:108 N-glycan profiles in H9N2 avian influenza viruses from chicken eggs and human embryonic lung fibroblast cells. 28797655 1 94 theme viruses 201:207 arg1 infectivity 174:184 infectivity 174:184 infectivity 174:184 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 94 theme viruses 201:207 arg1 virulence 160:168 virulence 160:168 virulence 160:168 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 1 94 theme viruses 201:207 arg1 specificity 147:157 the host specificity 138:157 the host specificity 138:157 N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). 28797655 12 95 theme other 2007:2011 arg1 viruses 2019:2025 other human viruses 2007:2025 other human viruses 2007:2025 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 5 96 theme H9N2 705:708 arg1 viruses 710:716 H9N2 viruses 705:716 H9N2 viruses 705:716 As a result, HA and NA of H9N2 viruses prossess six and five highly conserved N-glycosylation sites, respectively. 28797655 6 97 dep PHA-E 966:970 arg1 e.g. 960:963 e.g. 960:963 e.g. 960:963 Sixteen lectins (e.g., MAL-II, SNA and UEA-I) had increased expression levels of the glycan structures in the MRC-5 compared with the ECE system; however, 6 lectins (e.g., PHA-E, PSA and DSA) had contrasting results. 28797655 12 98 dep transmission 1844:1855 arg1 the 1834:1836 the 1834:1836 the 1834:1836 Our findings may provide a novel way for further discussing the molecular mechanism of the viral transmission and virulence associated with viral glycosylation in avian and human hosts as well as vital information for designing a vaccine against influenza and other human viruses. 28797655 11 99 theme glycan 1677:1682 arg1 structures 1684:1693 the exact glycan structures 1667:1693 the exact glycan structures for the same influenza strain from different hosts 1667:1744 Alterations in the glycosylation sites for the evolution and role of IAVs have been widely described; however, little is known about the exact glycan structures for the same influenza strain from different hosts. 28797655 2 100 dep distribution 236:247 arg1 the 232:234 the 232:234 the 232:234 In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. 28797655 10 101 theme existing 1394:1401 arg1 sialoglycans 1415:1426 the existing SAα2-3/6Gal sialoglycans 1390:1426 the existing SAα2-3/6Gal sialoglycans 1390:1426 We conclude that the existing SAα2-3/6Gal sialoglycans make the offspring of the H9N2 virus prefer entially attach to each other, which decreases the virulence. 28797655 3 102 theme chicken 468:474 arg1 ECE 482:484 ECE 482:484 ECE 482:484 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 28797655 3 102 theme chicken 468:474 arg1 eggs 476:479 embryonated chicken eggs 456:479 embryonated chicken eggs (ECE) 456:485 Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. 26059692 8 0 theme glycosylations 1206:1219 arg1 contribution 1176:1187 the contribution 1172:1187 the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1 1172:1309 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 9 1 theme Clostridium 1351:1361 arg1 perfringens 1363:1373 Clostridium perfringens 1351:1373 Clostridium perfringens 1351:1373 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 7 2 theme hierarchical 1023:1034 arg1 order 1036:1040 a hierarchical order 1021:1040 a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites 1021:1115 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 4 3 theme O-glycans 696:704 arg1 form 657:660 the form 653:660 the form of disialylated core 1 and core 2 O-glycans 653:704 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 5 4 theme chymotryptic 747:758 arg1 ETGATEKPTVIDSTIQSEFPTY 774:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 8 5 from contribution 1176:1187 arg1 oligomerization 1237:1251 neuropilin-1 oligomerization 1224:1251 neuropilin-1 oligomerization 1224:1251 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 8 5 from contribution 1176:1187 arg1 binding 1261:1267 the binding 1257:1267 the binding to partner proteins as VEGF or galectin-1 1257:1309 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 8 6 theme described 1196:1204 arg1 glycosylations 1206:1219 the described glycosylations 1192:1219 the described glycosylations 1192:1219 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 4 7 gly disialylated 665:676 arg1 core 678:681 disialylated core 1 and core 2 O-glycans 665:704 core 678:681 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 3 8 from components 527:536 arg1 surface 548:554 a cell surface 541:554 a cell surface 541:554 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 7 9 gly glycosylation 1097:1109 arg2 sites 1111:1115 the four different glycosylation sites 1078:1115 the four different glycosylation sites 1078:1115 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 7 9 gly glycosylation 1097:1109 arg2 four 1082:1085 four 1082:1085 four 1082:1085 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 5 10 theme c/MAM 760:764 arg1 ETGATEKPTVIDSTIQSEFPTY 774:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 3 11 theme neuropilin-1 365:376 arg1 domain 349:354 The c/MAM domain 339:354 The c/MAM domain of human neuropilin-1 339:376 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 3 11 theme neuropilin-1 365:376 arg1 neuropilin-1 365:376 human neuropilin-1 359:376 human neuropilin-1 359:376 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 3 12 from protein 503:509 arg1 surface 548:554 a cell surface 541:554 a cell surface 541:554 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 4 13 from O-glycosylated 600:613 arg1 form 657:660 the form 653:660 the form of disialylated core 1 and core 2 O-glycans 653:704 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 14 dep 24 626:627 arg1 to 623:624 to 623:624 to 623:624 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 2 15 theme neuropilin 285:294 arg1 oligomerization 296:310 neuropilin oligomerization 285:310 neuropilin oligomerization required for its function 285:336 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 6 16 gly glycosylated 873:884 arg1 species 886:892 highly glycosylated species 866:892 highly glycosylated species consisting of carbohydrate to about 50 % 866:933 For highly glycosylated species consisting of carbohydrate to about 50 %, useful results could only be obtained upon partial desialylation. 26059692 9 17 from perfringens 1363:1373 arg1 sialidase 1336:1344 the sialidase 1332:1344 the sialidase from Clostridium perfringens 1332:1373 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 2 18 theme proximal 170:177 arg1 neuropilin-1 189:200 neuropilin-1 189:200 neuropilin-1 189:200 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 2 18 theme proximal 170:177 arg1 domain 179:184 The membrane proximal domain 157:184 The membrane proximal domain 157:184 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 4 19 theme disialylated 665:676 arg1 core 678:681 disialylated core 1 and core 2 O-glycans 665:704 core 678:681 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 3 20 theme human 359:363 arg1 neuropilin-1 365:376 human neuropilin-1 359:376 human neuropilin-1 359:376 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 2 21 theme membrane 161:168 arg1 neuropilin-1 189:200 neuropilin-1 189:200 neuropilin-1 189:200 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 2 21 theme membrane 161:168 arg1 domain 179:184 The membrane proximal domain 157:184 The membrane proximal domain 157:184 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 5 22 theme electron-transfer 800:816 arg1 ETD-MS/MS 850:858 ETD-MS/MS 850:858 ETD-MS/MS 850:858 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 5 22 theme electron-transfer 800:816 arg1 spectrometry 836:847 electron-transfer dissociation mass spectrometry 800:847 electron-transfer dissociation mass spectrometry (ETD-MS/MS) 800:859 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 8 23 theme functional 1147:1156 arg1 studies 1158:1164 future functional studies 1140:1164 future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1 1140:1309 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 4 24 theme c/MAM 575:579 arg1 O-glycosylated 600:613 O-glycosylated 600:613 O-glycosylated 600:613 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 24 theme c/MAM 575:579 arg1 domain 581:586 the c/MAM domain 571:586 the c/MAM domain 571:586 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 3 25 theme c/MAM 343:347 arg1 domain 349:354 The c/MAM domain 339:354 The c/MAM domain of human neuropilin-1 339:376 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 3 25 theme c/MAM 343:347 arg1 neuropilin-1 365:376 human neuropilin-1 359:376 human neuropilin-1 359:376 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 0 26 theme Detailed 0:7 arg1 characterization 9:24 Detailed characterization 0:24 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 0:88 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 9 27 theme galactose- 1410:1419 arg1 acid 1461:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 4 28 gly O-glycosylated 600:613 arg2 domain 581:586 the c/MAM domain 571:586 the c/MAM domain 571:586 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 28 gly O-glycosylated 600:613 arg1 O-glycosylated 600:613 O-glycosylated 600:613 O-glycosylated 600:613 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 28 gly O-glycosylated 600:613 arg2 O-glycosylated 600:613 O-glycosylated 600:613 O-glycosylated 600:613 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 28 gly O-glycosylated 600:613 arg1 form 657:660 the form 653:660 the form of disialylated core 1 and core 2 O-glycans 653:704 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 28 gly O-glycosylated 600:613 arg1 domain 581:586 the c/MAM domain 571:586 the c/MAM domain 571:586 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 29 from form 657:660 arg1 O-glycosylated 600:613 O-glycosylated 600:613 O-glycosylated 600:613 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 29 from form 657:660 arg1 domain 581:586 the c/MAM domain 571:586 the c/MAM domain 571:586 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 3 30 theme propensity 445:454 arg1 investigation 424:436 investigation 424:436 investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface 424:554 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 8 31 theme partner 1272:1278 arg1 proteins 1280:1287 partner proteins 1272:1287 partner proteins as VEGF or galectin-1 1272:1309 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 0 32 link O-linked 33:40 arg1 glycosylation 42:54 the O-linked glycosylation 29:54 the O-linked glycosylation of the neuropilin-1 c/MAM-domain 29:87 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 7 33 theme glycosylation 1097:1109 arg1 sites 1111:1115 the four different glycosylation sites 1078:1115 the four different glycosylation sites 1078:1115 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 5 34 theme Attachment 707:716 arg1 sites 718:722 Attachment sites 707:722 Attachment sites 707:722 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 0 35 theme glycosylation 42:54 arg1 characterization 9:24 Detailed characterization 0:24 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 0:88 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 3 36 theme carbohydrate 514:525 arg1 components 527:536 other protein or carbohydrate components 497:536 components 527:536 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 3 37 with interactions 479:490 arg1 protein 503:509 other protein or carbohydrate components 497:536 protein 503:509 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 3 37 with interactions 479:490 arg1 components 527:536 other protein or carbohydrate components 497:536 components 527:536 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 2 38 theme sequence 239:246 arg1 conservation 248:259 its sequence conservation 235:259 its sequence conservation 235:259 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 4 39 theme core 678:681 arg1 form 657:660 the form 653:660 the form of disialylated core 1 and core 2 O-glycans 653:704 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 9 40 theme N-acetylgalactosamine-linked 1425:1452 arg1 acid 1461:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 0 41 theme O-linked 33:40 arg1 glycosylation 42:54 the O-linked glycosylation 29:54 the O-linked glycosylation of the neuropilin-1 c/MAM-domain 29:87 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 6 42 theme partial 979:985 arg1 desialylation 987:999 partial desialylation 979:999 partial desialylation 979:999 For highly glycosylated species consisting of carbohydrate to about 50 %, useful results could only be obtained upon partial desialylation. 26059692 5 43 theme dissociation 818:829 arg1 ETD-MS/MS 850:858 ETD-MS/MS 850:858 ETD-MS/MS 850:858 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 5 43 theme dissociation 818:829 arg1 spectrometry 836:847 electron-transfer dissociation mass spectrometry 800:847 electron-transfer dissociation mass spectrometry (ETD-MS/MS) 800:859 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 2 44 theme neuropilin-1 189:200 arg1 neuropilin-1 189:200 neuropilin-1 189:200 neuropilin-1 189:200 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 2 44 theme neuropilin-1 189:200 arg1 domain 179:184 The membrane proximal domain 157:184 The membrane proximal domain 157:184 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 7 45 theme O-GalNAc 1057:1064 arg1 addition 1066:1073 the initial O-GalNAc addition 1045:1073 the initial O-GalNAc addition 1045:1073 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 3 46 theme molecular 469:477 arg1 interactions 479:490 molecular interactions 469:490 molecular interactions with other protein or carbohydrate components on a cell surface 469:554 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 5 47 theme mass 831:834 arg1 ETD-MS/MS 850:858 ETD-MS/MS 850:858 ETD-MS/MS 850:858 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 5 47 theme mass 831:834 arg1 spectrometry 836:847 electron-transfer dissociation mass spectrometry 800:847 electron-transfer dissociation mass spectrometry (ETD-MS/MS) 800:859 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 8 48 theme neuropilin-1 1224:1235 arg1 oligomerization 1237:1251 neuropilin-1 oligomerization 1224:1251 neuropilin-1 oligomerization 1224:1251 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26059692 5 49 theme peptide 766:772 arg1 ETGATEKPTVIDSTIQSEFPTY 774:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY 743:795 Attachment sites were identified on the chymotryptic c/MAM peptide ETGATEKPTVIDSTIQSEFPTY by electron-transfer dissociation mass spectrometry (ETD-MS/MS). 26059692 7 50 theme addition 1066:1073 arg1 order 1036:1040 a hierarchical order 1021:1040 a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites 1021:1115 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 6 51 theme useful 936:941 arg1 results 943:949 useful results 936:949 useful results 936:949 For highly glycosylated species consisting of carbohydrate to about 50 %, useful results could only be obtained upon partial desialylation. 26059692 7 52 theme different 1087:1095 arg1 sites 1111:1115 the four different glycosylation sites 1078:1115 the four different glycosylation sites 1078:1115 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 9 53 theme spin-off 1316:1323 arg1 result 1325:1330 a spin-off result 1314:1330 a spin-off result 1314:1330 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 4 54 with O-glycosylated 600:613 arg1 units 644:648 up to 24 monosaccharide units 620:648 up to 24 monosaccharide units 620:648 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 1 55 theme neuronal 135:142 arg1 development 144:154 neuronal development 135:154 neuronal development 135:154 Neuropilins are involved in angiogenesis and neuronal development. 26059692 6 56 theme glycosylated 873:884 arg1 species 886:892 highly glycosylated species 866:892 highly glycosylated species consisting of carbohydrate to about 50 % 866:933 For highly glycosylated species consisting of carbohydrate to about 50 %, useful results could only be obtained upon partial desialylation. 26059692 2 57 theme MAM 215:217 arg1 domain 219:224 c or MAM domain 210:224 domain 219:224 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 9 58 theme sialic 1454:1459 arg1 acid 1461:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 7 59 theme initial 1049:1055 arg1 addition 1066:1073 the initial O-GalNAc addition 1045:1073 the initial O-GalNAc addition 1045:1073 ETD-MS/MS revealed a hierarchical order of the initial O-GalNAc addition to the four different glycosylation sites. 26059692 3 60 theme other 497:501 arg1 protein 503:509 other protein or carbohydrate components 497:536 protein 503:509 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 0 61 gly glycosylation 42:54 arg1 c/MAM-domain 76:87 the neuropilin-1 c/MAM-domain 59:87 the neuropilin-1 c/MAM-domain 59:87 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 9 62 link N-acetylgalactosamine-linked 1425:1452 arg1 acid 1461:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 galactose- and N-acetylgalactosamine-linked sialic acid 1410:1464 As a spin-off result the sialidase from Clostridium perfringens turned out to discriminate between galactose- and N-acetylgalactosamine-linked sialic acid. 26059692 0 63 theme c/MAM-domain 76:87 arg1 glycosylation 42:54 the O-linked glycosylation 29:54 the O-linked glycosylation of the neuropilin-1 c/MAM-domain 29:87 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 2 64 theme c 210:210 arg1 domain 219:224 c or MAM domain 210:224 domain 219:224 The membrane proximal domain of neuropilin-1, called c or MAM domain based on its sequence conservation, has been implicated in neuropilin oligomerization required for its function. 26059692 4 65 theme core 689:692 arg1 O-glycans 696:704 disialylated core 1 and core 2 O-glycans 665:704 O-glycans 696:704 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 4 66 theme monosaccharide 629:642 arg1 units 644:648 up to 24 monosaccharide units 620:648 up to 24 monosaccharide units 620:648 We found that the c/MAM domain was heavily O-glycosylated with up to 24 monosaccharide units in the form of disialylated core 1 and core 2 O-glycans. 26059692 0 67 theme neuropilin-1 63:74 arg1 c/MAM-domain 76:87 the neuropilin-1 c/MAM-domain 59:87 the neuropilin-1 c/MAM-domain 59:87 Detailed characterization of the O-linked glycosylation of the neuropilin-1 c/MAM-domain. 26059692 3 68 theme cell 543:546 arg1 surface 548:554 a cell surface 541:554 a cell surface 541:554 The c/MAM domain of human neuropilin-1 has been recombinantly expressed to allow for investigation of its propensity to engage in molecular interactions with other protein or carbohydrate components on a cell surface. 26059692 8 69 theme future 1140:1145 arg1 studies 1158:1164 future functional studies 1140:1164 future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1 1140:1309 These findings enable future functional studies about the contribution of the described glycosylations in neuropilin-1 oligomerization and the binding to partner proteins as VEGF or galectin-1. 26823463 7 0 theme binding 1230:1236 arg1 subsites 1238:1245 at least four binding subsites 1216:1245 at least four binding subsites 1216:1245 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 4 1 theme terminus 740:747 arg1 extension 721:729 a long extension 714:729 the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer 670:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 2 2 theme FOS 410:412 arg1 synthesis 397:405 the synthesis 393:405 the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut 393:486 It also catalyzes the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut. 26823463 6 3 theme GH32 1028:1031 arg1 enzymes 1039:1045 GH32 yeast enzymes 1028:1045 GH32 yeast enzymes having the traits observed in XdINV 1028:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 7 4 theme D80A 1097:1100 arg1 mutant 1102:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 0 5 from Analysis 11:18 arg1 Xanthophyllomyces 49:65 Xanthophyllomyces 49:65 Xanthophyllomyces 49:65 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 7 6 theme relevant 1143:1150 arg1 substrates 1152:1161 relevant substrates 1143:1161 relevant substrates 1143:1161 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 4 7 theme C 738:738 arg1 terminus 740:747 its C terminus 734:747 its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer 734:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 9 8 theme efficient 1610:1618 arg1 activity 1635:1642 the efficient fructosylating activity 1606:1642 the efficient fructosylating activity of XdINV on α-glucosides 1606:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 9 9 theme fructosylating 1620:1633 arg1 activity 1635:1642 the efficient fructosylating activity 1606:1642 the efficient fructosylating activity of XdINV on α-glucosides 1606:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 6 10 located observed 1065:1072 arg2 traits 1058:1063 the traits 1054:1063 the traits observed in XdINV 1054:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 6 10 located observed 1065:1072 arg1 XdINV 1077:1081 XdINV 1077:1081 XdINV 1077:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 10 11 theme residues 1691:1698 arg1 role 1674:1677 The role 1670:1677 The role of relevant residues 1670:1698 The role of relevant residues has been investigated by mutagenesis and kinetics measurements, and a model for the transfructosylating reaction has been proposed. 26823463 11 12 theme novel 1925:1929 arg1 bioconjugates 1931:1943 novel bioconjugates 1925:1943 novel bioconjugates 1925:1943 The plasticity of its active site makes XdINV a valuable and flexible biocatalyst to produce novel bioconjugates. 26823463 3 13 theme interesting 575:585 arg1 target 601:606 an interesting biotechnology target 572:606 it an interesting biotechnology target 569:606 In contrast to most fructosylating enzymes, XdINV produces neo-FOS, which makes it an interesting biotechnology target. 26823463 8 14 theme flexible 1371:1378 arg1 Glu-334-His-343 1386:1400 Glu-334-His-343 1386:1400 Glu-334-His-343 1386:1400 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 8 14 theme flexible 1371:1378 arg1 essential 1406:1414 essential 1406:1414 essential 1406:1414 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 8 14 theme flexible 1371:1378 arg1 loop 1380:1383 a flexible loop 1369:1383 a flexible loop (Glu-334-His-343) 1369:1401 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 6 15 theme group 1019:1023 arg1 representative 999:1012 representative 999:1012 representative 999:1012 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 0 16 theme Crucial 111:117 arg1 Role 119:122 the Crucial Role 107:122 the Crucial Role of N-Glycosylation in Oligomerization and Activity 107:173 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 4 17 theme expected 674:681 arg1 arrangement 693:703 the expected bimodular arrangement 670:703 the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer 670:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 1 18 from fructans 365:372 arg1 fructose 310:317 fructose 310:317 fructose from various fructooligosaccharides (FOS) and fructans 310:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 3 19 theme biotechnology 587:599 arg1 target 601:606 an interesting biotechnology target 572:606 it an interesting biotechnology target 569:606 In contrast to most fructosylating enzymes, XdINV produces neo-FOS, which makes it an interesting biotechnology target. 26823463 10 20 theme relevant 1682:1689 arg1 residues 1691:1698 relevant residues 1682:1698 relevant residues 1682:1698 The role of relevant residues has been investigated by mutagenesis and kinetics measurements, and a model for the transfructosylating reaction has been proposed. 26823463 1 21 theme various 324:330 arg1 FOS 356:358 FOS 356:358 FOS 356:358 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 21 theme various 324:330 arg1 fructooligosaccharides 332:353 various fructooligosaccharides 324:353 various fructooligosaccharides (FOS) 324:359 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 0 22 theme N-Glycosylation 127:141 arg1 Role 119:122 the Crucial Role 107:122 the Crucial Role of N-Glycosylation in Oligomerization and Activity 107:173 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 0 22 theme N-Glycosylation 127:141 arg1 Features 94:101 Unique Features 87:101 Unique Features 87:101 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 3 23 dep it 569:570 arg1 target 601:606 an interesting biotechnology target 572:606 it an interesting biotechnology target 569:606 In contrast to most fructosylating enzymes, XdINV produces neo-FOS, which makes it an interesting biotechnology target. 26823463 1 24 gly glycosylated 245:256 arg1 dendrorhousβ-fructofuranosidase 194:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase 176:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV) 176:232 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 24 gly glycosylated 245:256 arg1 enzyme 266:271 a highly glycosylated dimeric enzyme 236:271 a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans 236:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 2 25 theme bacteria 466:473 arg1 growth 445:450 the growth 441:450 the growth of beneficial bacteria in human gut 441:486 It also catalyzes the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut. 26823463 4 26 theme dimer 822:826 arg1 formation 798:806 the formation 794:806 the formation of an unusual dimer 794:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 5 27 theme long 882:885 arg1 crevice 887:893 a long crevice 880:893 a long crevice 880:893 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 4 28 theme three-dimensional 629:645 arg1 structure 647:655 its three-dimensional structure 625:655 its three-dimensional structure 625:655 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 0 29 from Xanthophyllomyces 49:65 arg1 β-Fructofuranosidase 23:42 β-Fructofuranosidase 23:42 β-Fructofuranosidase from Xanthophyllomyces dendrorhous 23:77 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 0 29 from Xanthophyllomyces 49:65 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous 0:77 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 4 30 theme unusual 814:820 arg1 dimer 822:826 an unusual dimer 811:826 an unusual dimer 811:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 7 31 theme crystals 1188:1195 arg1 structures 1197:1206 their crystals structures 1182:1206 their crystals structures showing at least four binding subsites at each active site 1182:1265 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 1 32 from fructooligosaccharides 332:353 arg1 fructose 310:317 fructose 310:317 fructose from various fructooligosaccharides (FOS) and fructans 310:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 0 33 theme Structural 0:9 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous 0:77 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 9 34 theme neokestose 1591:1600 arg1 activity 1635:1642 the efficient fructosylating activity 1606:1642 the efficient fructosylating activity of XdINV on α-glucosides 1606:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 9 34 theme neokestose 1591:1600 arg1 production 1577:1586 the production 1573:1586 the production of neokestose 1573:1600 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 9 35 theme XdINV 1647:1651 arg1 activity 1635:1642 the efficient fructosylating activity 1606:1642 the efficient fructosylating activity of XdINV on α-glucosides 1606:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 9 35 theme XdINV 1647:1651 arg1 production 1577:1586 the production 1573:1586 the production of neokestose 1573:1600 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 4 36 theme long 716:719 arg1 extension 721:729 a long extension 714:729 the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer 670:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 0 37 theme β-Fructofuranosidase 23:42 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous 0:77 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 7 38 theme inactive 1088:1095 arg1 mutant 1102:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 11 39 theme active 1854:1859 arg1 site 1861:1864 its active site 1850:1864 its active site 1850:1864 The plasticity of its active site makes XdINV a valuable and flexible biocatalyst to produce novel bioconjugates. 26823463 9 40 from production 1577:1586 arg1 α-glucosides 1656:1667 α-glucosides 1656:1667 α-glucosides 1656:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 3 41 theme most 504:507 arg1 enzymes 524:530 most fructosylating enzymes 504:530 most fructosylating enzymes 504:530 In contrast to most fructosylating enzymes, XdINV produces neo-FOS, which makes it an interesting biotechnology target. 26823463 2 42 theme human 478:482 arg1 gut 484:486 human gut 478:486 human gut 478:486 It also catalyzes the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut. 26823463 6 43 contain having 1047:1052 arg1 enzymes 1039:1045 GH32 yeast enzymes 1028:1045 GH32 yeast enzymes having the traits observed in XdINV 1028:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 6 43 contain having 1047:1052 arg2 traits 1058:1063 the traits 1054:1063 the traits observed in XdINV 1054:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 8 44 theme β 1439:1439 arg1 oligosaccharides 1453:1468 β(2-1)-linked oligosaccharides 1439:1468 β(2-1)-linked oligosaccharides 1439:1468 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 5 45 theme potential 921:929 arg1 ability 931:937 its potential ability to accommodate branched fructans 917:970 its potential ability to accommodate branched fructans 917:970 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 9 46 theme neo-type 1494:1501 arg1 substrates 1503:1512 neo-type substrates 1494:1512 neo-type substrates 1494:1512 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 1 47 theme glycosylated 245:256 arg1 dendrorhousβ-fructofuranosidase 194:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase 176:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV) 176:232 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 47 theme glycosylated 245:256 arg1 enzyme 266:271 a highly glycosylated dimeric enzyme 236:271 a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans 236:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 48 theme dimeric 258:264 arg1 dendrorhousβ-fructofuranosidase 194:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase 176:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV) 176:232 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 48 theme dimeric 258:264 arg1 enzyme 266:271 a highly glycosylated dimeric enzyme 236:271 a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans 236:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 10 49 theme transfructosylating 1784:1802 arg1 reaction 1804:1811 the transfructosylating reaction 1780:1811 the transfructosylating reaction 1780:1811 The role of relevant residues has been investigated by mutagenesis and kinetics measurements, and a model for the transfructosylating reaction has been proposed. 26823463 2 50 theme beneficial 455:464 arg1 bacteria 466:473 beneficial bacteria 455:473 beneficial bacteria 455:473 It also catalyzes the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut. 26823463 11 51 theme valuable 1880:1887 arg1 biocatalyst 1902:1912 a valuable and flexible biocatalyst 1878:1912 a valuable and flexible biocatalyst to produce novel bioconjugates 1878:1943 The plasticity of its active site makes XdINV a valuable and flexible biocatalyst to produce novel bioconjugates. 26823463 9 52 from activity 1635:1642 arg1 α-glucosides 1656:1667 α-glucosides 1656:1667 α-glucosides 1656:1667 Conversely, β(2-6) and neo-type substrates are accommodated mainly by stacking to Trp-105, explaining the production of neokestose and the efficient fructosylating activity of XdINV on α-glucosides. 26823463 0 53 from Features 94:101 arg1 Activity 166:173 Activity 166:173 Activity 166:173 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 0 53 from Features 94:101 arg1 Oligomerization 146:160 Oligomerization 146:160 Oligomerization 146:160 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 7 54 used used 1113:1116 arg2 mutant 1102:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant 1084:1107 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 11 55 theme site 1861:1864 arg1 plasticity 1836:1845 The plasticity 1832:1845 The plasticity of its active site 1832:1864 The plasticity of its active site makes XdINV a valuable and flexible biocatalyst to produce novel bioconjugates. 26823463 0 56 dep Xanthophyllomyces 49:65 arg1 dendrorhous 67:77 Xanthophyllomyces dendrorhous 49:77 Xanthophyllomyces dendrorhous 49:77 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 3 57 theme fructosylating 509:522 arg1 enzymes 524:530 most fructosylating enzymes 504:530 most fructosylating enzymes 504:530 In contrast to most fructosylating enzymes, XdINV produces neo-FOS, which makes it an interesting biotechnology target. 26823463 5 58 theme active 837:842 arg1 sites 844:848 The two active sites 829:848 The two active sites of the dimer 829:861 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 5 58 theme active 837:842 arg1 dimer 857:861 the dimer 853:861 the dimer 853:861 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 8 59 theme -linked 1445:1451 arg1 oligosaccharides 1453:1468 β(2-1)-linked oligosaccharides 1439:1468 β(2-1)-linked oligosaccharides 1439:1468 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 2 60 from growth 445:450 arg1 gut 484:486 human gut 478:486 human gut 478:486 It also catalyzes the synthesis of FOS, prebiotics that stimulate the growth of beneficial bacteria in human gut. 26823463 8 61 theme 2-1 1441:1443 arg1 oligosaccharides 1453:1468 β(2-1)-linked oligosaccharides 1439:1468 β(2-1)-linked oligosaccharides 1439:1468 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 11 62 theme flexible 1893:1900 arg1 biocatalyst 1902:1912 a valuable and flexible biocatalyst 1878:1912 a valuable and flexible biocatalyst to produce novel bioconjugates 1878:1943 The plasticity of its active site makes XdINV a valuable and flexible biocatalyst to produce novel bioconjugates. 26823463 4 63 theme anN-linked 768:777 arg1 glycan 779:784 anN-linked glycan 768:784 anN-linked glycan 768:784 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 8 64 theme different 1282:1290 arg1 positions 1292:1300 two different positions 1278:1300 two different positions 1278:1300 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 0 65 from Role 119:122 arg1 Activity 166:173 Activity 166:173 Activity 166:173 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 0 65 from Role 119:122 arg1 Oligomerization 146:160 Oligomerization 146:160 Oligomerization 146:160 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 4 66 link anN-linked 768:777 arg1 glycan 779:784 anN-linked glycan 768:784 anN-linked glycan 768:784 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 5 67 theme branched 954:961 arg1 fructans 963:970 branched fructans 954:970 branched fructans 954:970 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 8 68 theme subsite 1320:1326 arg1 +2 1328:1329 subsite +2 1320:1329 subsite +2 1320:1329 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 7 69 with complexes 1128:1136 arg1 products 1167:1174 products 1167:1174 products 1167:1174 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 7 69 with complexes 1128:1136 arg1 substrates 1152:1161 relevant substrates 1143:1161 relevant substrates 1143:1161 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 4 70 theme bimodular 683:691 arg1 arrangement 693:703 the expected bimodular arrangement 670:703 the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer 670:826 We present here its three-dimensional structure, which shows the expected bimodular arrangement and also a long extension of its C terminus that together with anN-linked glycan mediate the formation of an unusual dimer. 26823463 10 71 dep mutagenesis 1725:1735 arg1 measurements 1750:1761 measurements 1750:1761 measurements 1750:1761 The role of relevant residues has been investigated by mutagenesis and kinetics measurements, and a model for the transfructosylating reaction has been proposed. 26823463 6 72 theme enzymes 1039:1045 arg1 enzymes 1039:1045 GH32 yeast enzymes 1028:1045 GH32 yeast enzymes having the traits observed in XdINV 1028:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 6 72 theme enzymes 1039:1045 arg1 group 1019:1023 a group 1017:1023 a group of GH32 yeast enzymes having the traits observed in XdINV 1017:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 26823463 0 73 theme Unique 87:92 arg1 Features 94:101 Unique Features 87:101 Unique Features 87:101 Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-Glycosylation in Oligomerization and Activity. 26823463 5 74 theme dimer 857:861 arg1 sites 844:848 The two active sites 829:848 The two active sites of the dimer 829:861 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 5 74 theme dimer 857:861 arg1 dimer 857:861 the dimer 853:861 the dimer 853:861 The two active sites of the dimer are connected by a long crevice, which might indicate its potential ability to accommodate branched fructans. 26823463 8 75 link -linked 1445:1451 arg1 oligosaccharides 1453:1468 β(2-1)-linked oligosaccharides 1439:1468 β(2-1)-linked oligosaccharides 1439:1468 Moreover, two different positions are observed from subsite +2 depending on the substrate, and thus, a flexible loop (Glu-334-His-343) is essential in binding sucrose and β(2-1)-linked oligosaccharides. 26823463 1 76 theme Xanthophyllomyces 176:192 arg1 XdINV 227:231 XdINV 227:231 XdINV 227:231 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 76 theme Xanthophyllomyces 176:192 arg1 dendrorhousβ-fructofuranosidase 194:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase 176:224 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV) 176:232 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 1 76 theme Xanthophyllomyces 176:192 arg1 enzyme 266:271 a highly glycosylated dimeric enzyme 236:271 a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans 236:372 Xanthophyllomyces dendrorhousβ-fructofuranosidase (XdINV)is a highly glycosylated dimeric enzyme that hydrolyzes sucrose and releases fructose from various fructooligosaccharides (FOS) and fructans. 26823463 7 77 theme active 1255:1260 arg1 site 1262:1265 each active site 1250:1265 each active site 1250:1265 The inactive D80A mutant was used to obtain complexes with relevant substrates and products, with their crystals structures showing at least four binding subsites at each active site. 26823463 6 78 theme yeast 1033:1037 arg1 enzymes 1039:1045 GH32 yeast enzymes 1028:1045 GH32 yeast enzymes having the traits observed in XdINV 1028:1081 This arrangement could be representative of a group of GH32 yeast enzymes having the traits observed in XdINV. 28400175 12 0 theme IgE 1519:1521 arg1 antibodies 1523:1532 IgE antibodies 1519:1532 IgE antibodies 1519:1532 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 5 1 theme cell-derived 649:660 arg1 IgE 662:664 human cell-derived IgE 643:664 human cell-derived IgE 643:664 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 4 2 theme % 604:604 arg1 occupancy 606:614 100% occupancy 601:614 100% occupancy at GS1 and 2 601:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 2 3 theme N-glycosylation 289:303 arg1 pattern 305:311 the diverse N-glycosylation pattern 277:311 the diverse N-glycosylation pattern 277:311 However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. 28400175 13 4 with generation 1701:1710 arg1 N-glycans 1739:1747 other targeted N-glycans 1724:1747 other targeted N-glycans 1724:1747 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 6 5 theme adjacent 880:887 arg1 glycosites 889:898 adjacent glycosites 880:898 adjacent glycosites 880:898 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 6 6 theme spatial 792:798 arg1 position 800:807 spatial position 792:807 spatial position 792:807 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 6 7 from occupancy 867:875 arg1 glycosites 889:898 adjacent glycosites 880:898 adjacent glycosites 880:898 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 9 8 theme unmet 1232:1236 arg1 glycosylation 1208:1220 its glycosylation 1204:1220 its glycosylation 1204:1220 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 9 8 theme unmet 1232:1236 arg1 need 1238:1241 a so far unmet need 1223:1241 a so far unmet need 1223:1241 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 4 9 theme spectrometric 473:485 arg1 profiling 487:495 Mass spectrometric profiling 468:495 Mass spectrometric profiling 468:495 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 14 10 theme detailed 1771:1778 arg1 studies 1799:1805 detailed structure-function studies 1771:1805 detailed structure-function studies 1771:1805 This will facilitate detailed structure-function studies and may lead to the production of IgE with optimized activities. 28400175 4 11 gly glycosite 580:588 arg2 GS6 593:595 GS6 593:595 GS6 593:595 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 4 11 gly glycosite 580:588 arg2 glycosite 580:588 non-occupied at glycosite 6 564:590 non-occupied at glycosite 6 (GS6) 564:596 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 12 12 from impact 1549:1554 arg1 class 1590:1594 this antibody class 1576:1594 this antibody class 1576:1594 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 11 13 theme recombinant 1396:1406 arg1 proteins 1408:1415 recombinant proteins 1396:1415 recombinant proteins 1396:1415 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 9 14 theme well-established 1254:1269 arg1 systems 1282:1288 well-established expression systems 1254:1288 well-established expression systems 1254:1288 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 13 15 theme glyco-engineer 1628:1641 arg1 IgE 1655:1657 glyco-engineer recombinant IgE 1628:1657 glyco-engineer recombinant IgE 1628:1657 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 5 16 theme complex 688:694 arg1 N-glycans 696:704 complex N-glycans 688:704 complex N-glycans 688:704 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 3 17 from expression 335:344 arg1 plants 422:427 glycan-engineered Nicotiana benthamiana plants 382:427 glycan-engineered Nicotiana benthamiana plants 382:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 3 17 from expression 335:344 arg1 type 373:376 wild type 368:376 wild type 368:376 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 11 18 gly glycosylation 1379:1391 arg1 proteins 1408:1415 recombinant proteins 1396:1415 recombinant proteins 1396:1415 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 6 19 gly glycosites 889:898 arg2 glycosites 889:898 adjacent glycosites 880:898 adjacent glycosites 880:898 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 4 20 from GS1 619:621 arg1 occupancy 606:614 100% occupancy 601:614 100% occupancy at GS1 and 2 601:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 2 21 theme molecule 244:251 arg1 complexity 226:235 the complexity 222:235 the complexity of the molecule 222:251 However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. 28400175 4 22 theme site 527:530 arg1 occupancy 532:540 site occupancy 527:540 a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2 525:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 11 23 from advantage 1432:1440 arg1 applications 1457:1468 therapeutic applications 1445:1468 therapeutic applications 1445:1468 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 7 24 theme GnGnXF 999:1004 arg1 structures 1006:1015 plant-typical GnGnXF structures 985:1015 plant-typical GnGnXF structures 985:1015 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 3 25 theme recombinant 349:359 arg1 IgE 361:363 recombinant IgE 349:363 recombinant IgE 349:363 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 4 26 theme plant 511:515 arg1 IgE 517:519 plant IgE 511:519 plant IgE 511:519 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 8 27 theme glycan 1063:1068 arg1 diversity 1070:1078 the glycan diversity 1059:1078 the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms 1059:1145 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 1 28 theme human 97:101 arg1 antibodies 107:116 human IgE antibodies 97:116 human IgE antibodies 97:116 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 3 29 theme glycan-engineered 382:398 arg1 plants 422:427 glycan-engineered Nicotiana benthamiana plants 382:427 glycan-engineered Nicotiana benthamiana plants 382:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 13 30 theme important 1674:1682 arg1 step 1684:1687 an important step 1671:1687 an important step towards the generation of IgE with other targeted N-glycans 1671:1747 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 3 31 theme N-glycosylation 442:456 arg1 analyses 458:465 in-depth N-glycosylation analyses 433:465 in-depth N-glycosylation analyses 433:465 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 0 32 theme Recombinant 0:10 arg1 glycoproteomics 36:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics. 28400175 7 33 theme wild 938:941 arg1 type 943:946 wild type 938:946 wild type 938:946 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 13 34 theme IgE 1715:1717 arg1 generation 1701:1710 the generation 1697:1710 the generation of IgE with other targeted N-glycans 1697:1747 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 10 35 theme carbohydrate-dependent 1332:1353 arg1 functions 1359:1367 possible carbohydrate-dependent IgE functions 1323:1367 possible carbohydrate-dependent IgE functions 1323:1367 This enables the elucidation of possible carbohydrate-dependent IgE functions. 28400175 0 36 theme human 26:30 arg1 glycoproteomics 36:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics. 28400175 8 37 theme cell-derived 1087:1098 arg1 IgE 1100:1102 HEK cell-derived IgE 1083:1102 HEK cell-derived IgE 1083:1102 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 1 38 theme advanced 126:133 arg1 systems 135:141 advanced systems 126:141 advanced systems which allow their proper expression 126:177 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 13 39 theme other 1724:1728 arg1 N-glycans 1739:1747 other targeted N-glycans 1724:1747 other targeted N-glycans 1724:1747 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 6 40 theme site 862:865 arg1 occupancy 867:875 site occupancy 862:875 site occupancy 862:875 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 3 41 theme benthamiana 410:420 arg1 plants 422:427 glycan-engineered Nicotiana benthamiana plants 382:427 glycan-engineered Nicotiana benthamiana plants 382:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 5 42 contain carry 682:686 arg3 structures 735:744 GS1-5 and oligomannosidic structures 709:744 GS1-5 and oligomannosidic structures at GS7 709:751 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 5 42 contain carry 682:686 arg1 versions 673:680 plant versions 667:680 plant versions 667:680 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 5 42 contain carry 682:686 arg2 N-glycans 696:704 complex N-glycans 688:704 complex N-glycans 688:704 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 4 43 contain has 521:523 arg2 rate 542:545 a site occupancy rate 525:545 a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2 525:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 4 43 contain has 521:523 arg1 IgE 517:519 plant IgE 511:519 plant IgE 511:519 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 14 44 theme optimized 1850:1858 arg1 activities 1860:1869 optimized activities 1850:1869 optimized activities 1850:1869 This will facilitate detailed structure-function studies and may lead to the production of IgE with optimized activities. 28400175 4 45 theme non-occupied 564:575 arg1 glycosite 580:588 non-occupied at glycosite 6 564:590 non-occupied at glycosite 6 (GS6) 564:596 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 4 45 theme non-occupied 564:575 arg1 GS6 593:595 GS6 593:595 GS6 593:595 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 2 46 theme due 215:217 arg1 challenge 205:213 a challenge 203:213 a challenge due to the complexity of the molecule 203:251 However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. 28400175 2 46 theme due 215:217 arg1 this 189:192 this 189:192 this 189:192 However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. 28400175 10 47 theme functions 1359:1367 arg1 elucidation 1308:1318 the elucidation 1304:1318 the elucidation of possible carbohydrate-dependent IgE functions 1304:1367 This enables the elucidation of possible carbohydrate-dependent IgE functions. 28400175 14 48 theme IgE 1841:1843 arg1 production 1827:1836 the production 1823:1836 the production of IgE with optimized activities 1823:1869 This will facilitate detailed structure-function studies and may lead to the production of IgE with optimized activities. 28400175 12 49 theme glycosylation 1559:1571 arg1 scarce 1600:1605 scarce 1600:1605 scarce 1600:1605 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 12 49 theme glycosylation 1559:1571 arg1 impact 1549:1554 impact 1549:1554 impact of glycosylation on this antibody class 1549:1594 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 12 49 theme glycosylation 1559:1571 arg1 knowledge 1535:1543 knowledge 1535:1543 knowledge 1535:1543 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 3 50 from analyses 458:465 arg1 plants 422:427 glycan-engineered Nicotiana benthamiana plants 382:427 glycan-engineered Nicotiana benthamiana plants 382:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 3 50 from analyses 458:465 arg1 type 373:376 wild type 368:376 wild type 368:376 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 1 51 theme increasing 57:66 arg1 interest 85:92 The increasing biotechnological interest 53:92 The increasing biotechnological interest in human IgE antibodies 53:116 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 5 52 theme human 643:647 arg1 IgE 662:664 human cell-derived IgE 643:664 human cell-derived IgE 643:664 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 1 53 from interest 85:92 arg1 antibodies 107:116 human IgE antibodies 97:116 human IgE antibodies 97:116 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 12 54 from interest 1507:1514 arg1 antibodies 1523:1532 IgE antibodies 1519:1532 IgE antibodies 1519:1532 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 12 55 theme biotechnological 1490:1505 arg1 interest 1507:1514 biotechnological interest 1490:1514 biotechnological interest in IgE antibodies 1490:1532 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 0 56 link plant-derived 12:24 arg1 glycoproteomics 36:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics. 28400175 4 57 theme 100 601:603 arg1 % 604:604 % 604:604 % 604:604 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 11 58 theme therapeutic 1445:1455 arg1 applications 1457:1468 therapeutic applications 1445:1468 therapeutic applications 1445:1468 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 9 59 theme glycosylation 1208:1220 arg1 control 1193:1199 the control 1189:1199 the control of its glycosylation, a so far unmet need 1189:1241 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 2 60 theme diverse 281:287 arg1 pattern 305:311 the diverse N-glycosylation pattern 277:311 the diverse N-glycosylation pattern 277:311 However, this is still a challenge due to the complexity of the molecule, particularly regarding the diverse N-glycosylation pattern. 28400175 11 61 theme Targeted 1370:1377 arg1 glycosylation 1379:1391 Targeted glycosylation 1370:1391 Targeted glycosylation of recombinant proteins 1370:1415 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 13 62 theme recombinant 1643:1653 arg1 IgE 1655:1657 glyco-engineer recombinant IgE 1628:1657 glyco-engineer recombinant IgE 1628:1657 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 8 63 link cell-derived 1087:1098 arg1 IgE 1100:1102 HEK cell-derived IgE 1083:1102 HEK cell-derived IgE 1083:1102 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 7 64 contain carry 955:959 arg2 GnGn 976:979 GnGn 976:979 GnGn 976:979 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 7 64 contain carry 955:959 arg1 IgE 901:903 IgE 901:903 IgE expressed in glycoengineered and wild type plants 901:953 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 7 64 contain carry 955:959 arg2 structures 1006:1015 plant-typical GnGnXF structures 985:1015 plant-typical GnGnXF structures 985:1015 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 7 64 contain carry 955:959 arg3 homogeneity 1026:1036 large homogeneity 1020:1036 large homogeneity 1020:1036 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 11 65 theme proteins 1408:1415 arg1 glycosylation 1379:1391 Targeted glycosylation 1370:1391 Targeted glycosylation of recombinant proteins 1370:1415 Targeted glycosylation of recombinant proteins may provide an advantage in therapeutic applications. 28400175 5 66 theme GS1-5 709:713 arg1 structures 735:744 GS1-5 and oligomannosidic structures 709:744 GS1-5 and oligomannosidic structures at GS7 709:751 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 5 67 from GS7 749:751 arg1 structures 735:744 GS1-5 and oligomannosidic structures 709:744 GS1-5 and oligomannosidic structures at GS7 709:751 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 4 68 from 2 627:627 arg1 occupancy 606:614 100% occupancy 601:614 100% occupancy at GS1 and 2 601:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 6 69 from processing 848:857 arg1 glycosites 889:898 adjacent glycosites 880:898 adjacent glycosites 880:898 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 5 70 theme plant 667:671 arg1 versions 673:680 plant versions 667:680 plant versions 667:680 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 14 71 with production 1827:1836 arg1 activities 1860:1869 optimized activities 1850:1869 optimized activities 1850:1869 This will facilitate detailed structure-function studies and may lead to the production of IgE with optimized activities. 28400175 9 72 theme expression 1271:1280 arg1 systems 1282:1288 well-established expression systems 1254:1288 well-established expression systems 1254:1288 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 7 73 theme large 1020:1024 arg1 homogeneity 1026:1036 large homogeneity 1020:1036 large homogeneity 1020:1036 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 4 74 theme Mass 468:471 arg1 profiling 487:495 Mass spectrometric profiling 468:495 Mass spectrometric profiling 468:495 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 1 75 theme proper 161:166 arg1 expression 168:177 their proper expression 155:177 their proper expression 155:177 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 4 76 theme occupancy 532:540 arg1 rate 542:545 a site occupancy rate 525:545 a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2 525:627 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 7 77 theme plant-typical 985:997 arg1 structures 1006:1015 plant-typical GnGnXF structures 985:1015 plant-typical GnGnXF structures 985:1015 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 5 78 theme oligomannosidic 719:733 arg1 structures 735:744 GS1-5 and oligomannosidic structures 709:744 GS1-5 and oligomannosidic structures at GS7 709:751 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 1 79 theme biotechnological 68:83 arg1 interest 85:92 The increasing biotechnological interest 53:92 The increasing biotechnological interest in human IgE antibodies 53:116 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 3 80 theme wild 368:371 arg1 type 373:376 wild type 368:376 wild type 368:376 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 3 81 theme IgE 361:363 arg1 expression 335:344 the expression 331:344 the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants 331:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 3 81 theme IgE 361:363 arg1 analyses 458:465 in-depth N-glycosylation analyses 433:465 in-depth N-glycosylation analyses 433:465 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 0 82 theme plant-derived 12:24 arg1 glycoproteomics 36:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics. 28400175 8 83 theme HEK 1083:1085 arg1 IgE 1100:1102 HEK cell-derived IgE 1083:1102 HEK cell-derived IgE 1083:1102 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 1 84 theme IgE 103:105 arg1 antibodies 107:116 human IgE antibodies 97:116 human IgE antibodies 97:116 The increasing biotechnological interest in human IgE antibodies demands advanced systems which allow their proper expression. 28400175 3 85 theme Nicotiana 400:408 arg1 plants 422:427 glycan-engineered Nicotiana benthamiana plants 382:427 glycan-engineered Nicotiana benthamiana plants 382:427 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 10 86 theme IgE 1355:1357 arg1 functions 1359:1367 possible carbohydrate-dependent IgE functions 1323:1367 possible carbohydrate-dependent IgE functions 1323:1367 This enables the elucidation of possible carbohydrate-dependent IgE functions. 28400175 14 87 theme structure-function 1780:1797 arg1 studies 1799:1805 detailed structure-function studies 1771:1805 detailed structure-function studies 1771:1805 This will facilitate detailed structure-function studies and may lead to the production of IgE with optimized activities. 28400175 0 88 theme IgE 32:34 arg1 glycoproteomics 36:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics 0:50 Recombinant plant-derived human IgE glycoproteomics. 28400175 8 89 theme IgE 1100:1102 arg1 diversity 1070:1078 the glycan diversity 1059:1078 the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms 1059:1145 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 7 90 theme type 943:946 arg1 plants 948:953 glycoengineered and wild type plants 918:953 glycoengineered and wild type plants 918:953 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 10 91 theme possible 1323:1330 arg1 functions 1359:1367 possible carbohydrate-dependent IgE functions 1323:1367 possible carbohydrate-dependent IgE functions 1323:1367 This enables the elucidation of possible carbohydrate-dependent IgE functions. 28400175 3 92 theme in-depth 433:440 arg1 analyses 458:465 in-depth N-glycosylation analyses 433:465 in-depth N-glycosylation analyses 433:465 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 13 93 theme targeted 1730:1737 arg1 N-glycans 1739:1747 other targeted N-glycans 1724:1747 other targeted N-glycans 1724:1747 With the ability to glyco-engineer recombinant IgE, we provide an important step towards the generation of IgE with other targeted N-glycans. 28400175 5 94 link cell-derived 649:660 arg1 IgE 662:664 human cell-derived IgE 643:664 human cell-derived IgE 643:664 Similarly to human cell-derived IgE, plant versions carry complex N-glycans at GS1-5 and oligomannosidic structures at GS7. 28400175 12 95 theme antibody 1581:1588 arg1 class 1590:1594 this antibody class 1576:1594 this antibody class 1576:1594 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 4 96 theme at 577:578 arg1 glycosite 580:588 non-occupied at glycosite 6 564:590 non-occupied at glycosite 6 (GS6) 564:596 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 4 96 theme at 577:578 arg1 GS6 593:595 GS6 593:595 GS6 593:595 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 6 97 theme Computational 754:766 arg1 modelling 768:776 Computational modelling 754:776 Computational modelling 754:776 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 7 98 theme glycoengineered 918:932 arg1 plants 948:953 glycoengineered and wild type plants 918:953 glycoengineered and wild type plants 918:953 IgE expressed in glycoengineered and wild type plants carry, respectively, GnGn and plant-typical GnGnXF structures at large homogeneity. 28400175 8 99 theme different 1126:1134 arg1 glycoforms 1136:1145 at least 20 different glycoforms 1114:1145 at least 20 different glycoforms 1114:1145 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 4 100 from glycosite 580:588 arg1 at 577:578 non-occupied at glycosite 6 564:590 non-occupied at glycosite 6 (GS6) 564:596 Mass spectrometric profiling revealed that plant IgE has a site occupancy rate that ranges from non-occupied at glycosite 6 (GS6) to 100% occupancy at GS1 and 2. 28400175 9 101 theme IgE 1161:1163 arg1 glycoengineering 1165:1180 IgE glycoengineering 1161:1180 IgE glycoengineering 1161:1180 Importantly, IgE glycoengineering allows the control of its glycosylation, a so far unmet need when using well-established expression systems. 28400175 12 102 from knowledge 1535:1543 arg1 class 1590:1594 this antibody class 1576:1594 this antibody class 1576:1594 Despite increasing biotechnological interest in IgE antibodies, knowledge and impact of glycosylation on this antibody class are scarce. 28400175 6 103 theme glycans 829:835 arg1 position 800:807 spatial position 792:807 spatial position 792:807 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 6 103 theme glycans 829:835 arg1 orientation 813:823 orientation 813:823 orientation 813:823 Computational modelling suggests that spatial position (or orientation) of glycans can impair processing or site occupancy on adjacent glycosites. 28400175 3 104 gly N-glycosylation 442:456 arg1 IgE 361:363 recombinant IgE 349:363 recombinant IgE 349:363 Here, we present the expression of recombinant IgE in wild type and glycan-engineered Nicotiana benthamiana plants and in-depth N-glycosylation analyses. 28400175 8 105 contain carrying 1105:1112 arg2 glycoforms 1136:1145 at least 20 different glycoforms 1114:1145 at least 20 different glycoforms 1114:1145 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 28400175 8 105 contain carrying 1105:1112 arg1 IgE 1100:1102 HEK cell-derived IgE 1083:1102 HEK cell-derived IgE 1083:1102 This contrasts with the glycan diversity of HEK cell-derived IgE, carrying at least 20 different glycoforms. 24917611 1 0 theme multiple 266:273 arg1 infections 290:299 multiple drug resistant infections 266:299 multiple drug resistant infections 266:299 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 3 1 theme protein 566:572 arg1 glycosylation 574:586 O-linked protein glycosylation 557:586 O-linked protein glycosylation 557:586 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 1 2 theme resistant 280:288 arg1 infections 290:299 multiple drug resistant infections 266:299 multiple drug resistant infections 266:299 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 4 3 theme glycan 691:696 arg1 biogenesis 698:707 glycan biogenesis 691:707 glycan biogenesis 691:707 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 9 4 theme O-linked 1592:1599 arg1 glycosylation 1601:1613 Acinetobacter O-linked glycosylation 1578:1613 Acinetobacter O-linked glycosylation 1578:1613 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 6 5 theme A. 1039:1040 arg1 strains 1052:1058 different A. baumannii strains 1029:1058 different A. baumannii strains 1029:1058 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 7 6 link O-linked 1346:1353 arg1 glycosylation 1355:1367 Acinetobacter O-linked glycosylation 1332:1367 Acinetobacter O-linked glycosylation 1332:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 5 7 theme extensive 894:902 arg1 diversity 911:919 extensive glycan diversity 894:919 extensive glycan diversity 894:919 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 1 8 from frequency 253:261 arg1 settings 228:235 clinical settings 219:235 clinical settings 219:235 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 9 from persistence 204:214 arg1 settings 228:235 clinical settings 219:235 clinical settings 219:235 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 8 10 theme alternative 1418:1428 arg1 glycoforms 1430:1439 20 alternative glycoforms 1415:1439 20 alternative glycoforms characterized within the 15 strains 1415:1475 By comparing the 11 main glycoforms and over 20 alternative glycoforms characterized within the 15 strains, trends within the glycan utilized for O-linked glycosylation could be observed. 24917611 6 11 dep A. 1039:1040 arg1 baumannii 1042:1050 baumannii 1042:1050 baumannii 1042:1050 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 3 12 theme single 464:469 arg1 responsible 496:506 responsible 496:506 responsible 496:506 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 3 12 theme single 464:469 arg1 pathway 484:490 a single biosynthetic pathway 462:490 a single biosynthetic pathway 462:490 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 1 13 dep pathogen 110:117 arg1 baumannii 133:141 Acinetobacter baumannii 119:141 The opportunistic human pathogen Acinetobacter baumannii 86:141 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 7 14 theme O-linked 1346:1353 arg1 glycosylation 1355:1367 Acinetobacter O-linked glycosylation 1332:1367 Acinetobacter O-linked glycosylation 1332:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 5 15 theme mass 834:837 arg1 methods 852:858 global and targeted mass spectrometry methods 814:858 global and targeted mass spectrometry methods 814:858 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 1 16 theme health 159:164 arg1 worldwide 179:187 health care systems worldwide 159:187 health care systems worldwide 159:187 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 17 theme systems 171:177 arg1 worldwide 179:187 health care systems worldwide 159:187 health care systems worldwide 159:187 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 6 18 theme similar 1067:1073 arg1 both 1095:1098 both 1095:1098 both 1095:1098 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 18 theme similar 1067:1073 arg1 substrates 1083:1092 similar protein substrates 1067:1092 similar protein substrates 1067:1092 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 9 19 dep five 1638:1641 arg1 to 1635:1636 to 1635:1636 to 1635:1636 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 7 20 theme widespread 1308:1317 arg1 heterogeneity 1289:1301 glycan heterogeneity 1282:1301 glycan heterogeneity 1282:1301 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 7 20 theme widespread 1308:1317 arg1 phenomena 1319:1327 a widespread phenomena 1306:1327 a widespread phenomena in Acinetobacter O-linked glycosylation 1306:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 3 21 link O-linked 557:564 arg1 glycosylation 574:586 O-linked protein glycosylation 557:586 O-linked protein glycosylation 557:586 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 10 22 theme capsule/O-linked 1851:1866 arg1 pathways 1888:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 0 23 theme glycosylation 38:50 arg1 systems 52:58 the O-linked protein glycosylation systems 17:58 the O-linked protein glycosylation systems of acinetobacter species 17:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 9 24 theme charged 1705:1711 arg1 derivatives 1777:1787 legionaminic/pseudaminic acid derivatives 1747:1787 legionaminic/pseudaminic acid derivatives 1747:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 24 theme charged 1705:1711 arg1 sugars 1713:1718 negatively charged sugars 1694:1718 negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives 1694:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 24 theme charged 1705:1711 arg1 GlcNAc3NAcA4OAc 1728:1742 GlcNAc3NAcA4OAc 1728:1742 GlcNAc3NAcA4OAc 1728:1742 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 0 25 theme species 77:83 arg1 systems 52:58 the O-linked protein glycosylation systems 17:58 the O-linked protein glycosylation systems of acinetobacter species 17:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 2 26 theme environmental 391:403 arg1 persistence 405:415 environmental persistence 391:415 environmental persistence of A. baumannii 391:431 To combat this threat, it is necessary to understand factors associated with disease and environmental persistence of A. baumannii. 24917611 7 27 theme glycan 1282:1287 arg1 heterogeneity 1289:1301 glycan heterogeneity 1282:1301 glycan heterogeneity 1282:1301 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 7 27 theme glycan 1282:1287 arg1 phenomena 1319:1327 a widespread phenomena 1306:1327 a widespread phenomena in Acinetobacter O-linked glycosylation 1306:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 1 28 theme infections 290:299 arg1 frequency 253:261 the growing frequency 241:261 the growing frequency of multiple drug resistant infections 241:299 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 28 theme infections 290:299 arg1 persistence 204:214 its persistence 200:214 its persistence in clinical settings 200:235 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 29 theme human 104:108 arg1 concern 148:154 a concern 146:154 a concern to health care systems worldwide 146:187 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 29 theme human 104:108 arg1 pathogen 110:117 The opportunistic human pathogen 86:117 The opportunistic human pathogen Acinetobacter baumannii 86:141 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 7 30 located observed 1223:1230 arg2 micro-heterogeneity 1194:1212 glycan micro-heterogeneity 1187:1212 glycan micro-heterogeneity 1187:1212 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 7 30 located observed 1223:1230 arg1 isolates 1250:1257 nearly all isolates 1239:1257 nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation 1239:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 4 31 theme glycoproteome 789:801 arg1 composition 729:739 composition 729:739 composition 729:739 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 4 31 theme glycoproteome 789:801 arg1 diversity 742:750 diversity 742:750 diversity 742:750 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 4 31 theme glycoproteome 789:801 arg1 properties 757:766 properties 757:766 properties 757:766 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 10 32 theme glycan 1868:1873 arg1 pathways 1888:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 8 33 theme O-linked 1516:1523 arg1 glycosylation 1525:1537 O-linked glycosylation 1516:1537 O-linked glycosylation 1516:1537 By comparing the 11 main glycoforms and over 20 alternative glycoforms characterized within the 15 strains, trends within the glycan utilized for O-linked glycosylation could be observed. 24917611 7 34 theme all 1246:1248 arg1 isolates 1250:1257 nearly all isolates 1239:1257 nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation 1239:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 0 35 theme protein 30:36 arg1 systems 52:58 the O-linked protein glycosylation systems 17:58 the O-linked protein glycosylation systems of acinetobacter species 17:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 10 36 with glycans 1906:1912 arg1 characteristics 1927:1941 similar characteristics 1919:1941 similar characteristics across all A. baumannii 1919:1965 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 4 37 dep driven 674:679 arg1 non-template 661:672 non-template 661:672 non-template 661:672 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 0 38 theme acinetobacter 63:75 arg1 species 77:83 acinetobacter species 63:83 acinetobacter species 63:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 9 39 dep short 1622:1626 arg1 residue 1643:1649 three to five residue 1629:1649 three to five residue 1629:1649 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 40 theme legionaminic/pseudaminic 1747:1770 arg1 derivatives 1777:1787 legionaminic/pseudaminic acid derivatives 1747:1787 legionaminic/pseudaminic acid derivatives 1747:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 4 41 theme carbohydrates 625:637 arg1 nature 681:686 the non-template driven nature 657:686 the non-template driven nature of glycan biogenesis 657:707 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 4 41 theme carbohydrates 625:637 arg1 requirement 604:614 the requirement 600:614 the requirement of these carbohydrates for virulence 600:651 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 10 42 link capsule/O-linked 1851:1866 arg1 pathways 1888:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 1 43 theme growing 245:251 arg1 frequency 253:261 the growing frequency 241:261 the growing frequency of multiple drug resistant infections 241:299 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 9 44 contain containing 1683:1692 arg2 GlcNAc3NAcA4OAc 1728:1742 GlcNAc3NAcA4OAc 1728:1742 GlcNAc3NAcA4OAc 1728:1742 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 44 contain containing 1683:1692 arg1 branching 1673:1681 limited branching 1665:1681 limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives 1665:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 44 contain containing 1683:1692 arg2 derivatives 1777:1787 legionaminic/pseudaminic acid derivatives 1747:1787 legionaminic/pseudaminic acid derivatives 1747:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 44 contain containing 1683:1692 arg2 sugars 1713:1718 negatively charged sugars 1694:1718 negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives 1694:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 10 45 theme similar 1919:1925 arg1 characteristics 1927:1941 similar characteristics 1919:1941 similar characteristics across all A. baumannii 1919:1965 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 0 46 link O-linked 21:28 arg1 systems 52:58 the O-linked protein glycosylation systems 17:58 the O-linked protein glycosylation systems of acinetobacter species 17:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 6 47 theme identity 1163:1170 arg1 O-glycosylation 1135:1149 O-glycosylation 1135:1149 O-glycosylation 1135:1149 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 47 theme identity 1163:1170 arg1 identity 1163:1170 protein identity 1155:1170 protein identity 1155:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 47 theme identity 1163:1170 arg1 sites 1126:1130 the sites 1122:1130 the sites of O-glycosylation and protein identity 1122:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 7 48 theme glycan 1187:1192 arg1 micro-heterogeneity 1194:1212 glycan micro-heterogeneity 1187:1212 glycan micro-heterogeneity 1187:1212 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 3 49 theme O-linked 557:564 arg1 glycosylation 574:586 O-linked protein glycosylation 557:586 O-linked protein glycosylation 557:586 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 1 50 theme drug 275:278 arg1 infections 290:299 multiple drug resistant infections 266:299 multiple drug resistant infections 266:299 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 6 51 gly glycoproteins 990:1002 arg1 glycoproteins 990:1002 the 26 glycoproteins 983:1002 the 26 glycoproteins identified 983:1013 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 3 52 theme polysaccharide 538:551 arg1 generation 516:525 the generation 512:525 the generation of capsule polysaccharide and O-linked protein glycosylation 512:586 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 5 53 theme glycan 904:909 arg1 diversity 911:919 extensive glycan diversity 894:919 extensive glycan diversity 894:919 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 9 54 theme Acinetobacter 1578:1590 arg1 glycosylation 1601:1613 Acinetobacter O-linked glycosylation 1578:1613 Acinetobacter O-linked glycosylation 1578:1613 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 4 55 theme biogenesis 698:707 arg1 nature 681:686 the non-template driven nature 657:686 the non-template driven nature of glycan biogenesis 657:707 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 4 55 theme biogenesis 698:707 arg1 requirement 604:614 the requirement 600:614 the requirement of these carbohydrates for virulence 600:651 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 3 56 theme glycosylation 574:586 arg1 generation 516:525 the generation 512:525 the generation of capsule polysaccharide and O-linked protein glycosylation 512:586 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 6 57 theme different 1029:1037 arg1 strains 1052:1058 different A. baumannii strains 1029:1058 different A. baumannii strains 1029:1058 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 5 58 theme Acinetobacter 954:966 arg1 glycoproteome 937:949 the O-linked glycoproteome 924:949 the O-linked glycoproteome of Acinetobacter 924:966 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 9 59 theme limited 1665:1671 arg1 branching 1673:1681 limited branching 1665:1681 limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives 1665:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 60 theme short 1622:1626 arg1 glycans 1652:1658 short (three to five residue) glycans 1622:1658 short (three to five residue) glycans 1622:1658 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 8 61 theme main 1390:1393 arg1 glycoforms 1395:1404 the 11 main glycoforms 1383:1404 the 11 main glycoforms 1383:1404 By comparing the 11 main glycoforms and over 20 alternative glycoforms characterized within the 15 strains, trends within the glycan utilized for O-linked glycosylation could be observed. 24917611 6 62 theme glycoproteins 990:1002 arg1 Comparison 969:978 Comparison 969:978 Comparison of the 26 glycoproteins identified 969:1013 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 7 63 from phenomena 1319:1327 arg1 glycosylation 1355:1367 Acinetobacter O-linked glycosylation 1332:1367 Acinetobacter O-linked glycosylation 1332:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 7 64 theme Acinetobacter 1332:1344 arg1 glycosylation 1355:1367 Acinetobacter O-linked glycosylation 1332:1367 Acinetobacter O-linked glycosylation 1332:1367 Surprisingly, glycan micro-heterogeneity was also observed within nearly all isolates examined demonstrating glycan heterogeneity is a widespread phenomena in Acinetobacter O-linked glycosylation. 24917611 5 65 theme O-linked 928:935 arg1 glycoproteome 937:949 the O-linked glycoproteome 924:949 the O-linked glycoproteome of Acinetobacter 924:966 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 8 66 link O-linked 1516:1523 arg1 glycosylation 1525:1537 O-linked glycosylation 1516:1537 O-linked glycosylation 1516:1537 By comparing the 11 main glycoforms and over 20 alternative glycoforms characterized within the 15 strains, trends within the glycan utilized for O-linked glycosylation could be observed. 24917611 5 67 theme global 814:819 arg1 methods 852:858 global and targeted mass spectrometry methods 814:858 global and targeted mass spectrometry methods 814:858 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 1 68 theme care 166:169 arg1 worldwide 179:187 health care systems worldwide 159:187 health care systems worldwide 159:187 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 6 69 theme protein 1075:1081 arg1 both 1095:1098 both 1095:1098 both 1095:1098 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 69 theme protein 1075:1081 arg1 substrates 1083:1092 similar protein substrates 1067:1092 similar protein substrates 1067:1092 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 70 theme protein 1155:1161 arg1 identity 1163:1170 protein identity 1155:1170 protein identity 1155:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 1 71 theme opportunistic 90:102 arg1 concern 148:154 a concern 146:154 a concern to health care systems worldwide 146:187 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 1 71 theme opportunistic 90:102 arg1 pathogen 110:117 The opportunistic human pathogen 86:117 The opportunistic human pathogen Acinetobacter baumannii 86:141 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 6 72 theme O-glycosylation 1135:1149 arg1 O-glycosylation 1135:1149 O-glycosylation 1135:1149 O-glycosylation 1135:1149 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 72 theme O-glycosylation 1135:1149 arg1 identity 1163:1170 protein identity 1155:1170 protein identity 1155:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 72 theme O-glycosylation 1135:1149 arg1 sites 1126:1130 the sites 1122:1130 the sites of O-glycosylation and protein identity 1122:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 0 73 theme O-linked 21:28 arg1 systems 52:58 the O-linked protein glycosylation systems 17:58 the O-linked protein glycosylation systems of acinetobacter species 17:83 Diversity within the O-linked protein glycosylation systems of acinetobacter species. 24917611 6 74 theme sites 1126:1130 arg1 characteristics 1103:1117 characteristics 1103:1117 characteristics of the sites of O-glycosylation and protein identity 1103:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 4 75 theme Acinetobacter 775:787 arg1 glycoproteome 789:801 the Acinetobacter glycoproteome 771:801 the Acinetobacter glycoproteome 771:801 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 3 76 theme biosynthetic 471:482 arg1 responsible 496:506 responsible 496:506 responsible 496:506 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 3 76 theme biosynthetic 471:482 arg1 pathway 484:490 a single biosynthetic pathway 462:490 a single biosynthetic pathway 462:490 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24917611 5 77 theme targeted 825:832 arg1 methods 852:858 global and targeted mass spectrometry methods 814:858 global and targeted mass spectrometry methods 814:858 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 5 78 link O-linked 928:935 arg1 glycoproteome 937:949 the O-linked glycoproteome 924:949 the O-linked glycoproteome of Acinetobacter 924:966 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 1 79 theme clinical 219:226 arg1 settings 228:235 clinical settings 219:235 clinical settings 219:235 The opportunistic human pathogen Acinetobacter baumannii is a concern to health care systems worldwide because of its persistence in clinical settings and the growing frequency of multiple drug resistant infections. 24917611 2 80 theme A. 420:421 arg1 persistence 405:415 environmental persistence 391:415 environmental persistence of A. baumannii 391:431 To combat this threat, it is necessary to understand factors associated with disease and environmental persistence of A. baumannii. 24917611 2 80 theme A. 420:421 arg1 disease 379:385 disease 379:385 disease 379:385 To combat this threat, it is necessary to understand factors associated with disease and environmental persistence of A. baumannii. 24917611 5 81 theme spectrometry 839:850 arg1 methods 852:858 global and targeted mass spectrometry methods 814:858 global and targeted mass spectrometry methods 814:858 Utilizing global and targeted mass spectrometry methods, we examined 15 strains and found extensive glycan diversity in the O-linked glycoproteome of Acinetobacter. 24917611 9 82 theme acid 1772:1775 arg1 derivatives 1777:1787 legionaminic/pseudaminic acid derivatives 1747:1787 legionaminic/pseudaminic acid derivatives 1747:1787 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 9 83 link O-linked 1592:1599 arg1 glycosylation 1601:1613 Acinetobacter O-linked glycosylation 1578:1613 Acinetobacter O-linked glycosylation 1578:1613 These trends reveal Acinetobacter O-linked glycosylation favors short (three to five residue) glycans with limited branching containing negatively charged sugars such as GlcNAc3NAcA4OAc or legionaminic/pseudaminic acid derivatives. 24917611 6 84 gly O-glycosylation 1135:1149 arg2 identity 1163:1170 protein identity 1155:1170 protein identity 1155:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 84 gly O-glycosylation 1135:1149 arg2 O-glycosylation 1135:1149 O-glycosylation 1135:1149 O-glycosylation 1135:1149 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 6 84 gly O-glycosylation 1135:1149 arg2 sites 1126:1130 the sites 1122:1130 the sites of O-glycosylation and protein identity 1122:1170 Comparison of the 26 glycoproteins identified revealed that different A. baumannii strains target similar protein substrates, both in characteristics of the sites of O-glycosylation and protein identity. 24917611 10 85 theme biosynthetic 1875:1886 arg1 pathways 1888:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 the capsule/O-linked glycan biosynthetic pathways 1847:1895 These observations suggest that although highly diverse, the capsule/O-linked glycan biosynthetic pathways generate glycans with similar characteristics across all A. baumannii. 24917611 4 86 theme driven 674:679 arg1 nature 681:686 the non-template driven nature 657:686 the non-template driven nature of glycan biogenesis 657:707 Because of the requirement of these carbohydrates for virulence and the non-template driven nature of glycan biogenesis we investigated the composition, diversity, and properties of the Acinetobacter glycoproteome. 24917611 3 87 theme capsule 530:536 arg1 polysaccharide 538:551 capsule polysaccharide 530:551 capsule polysaccharide 530:551 Recently, it was shown that a single biosynthetic pathway was responsible for the generation of capsule polysaccharide and O-linked protein glycosylation. 24632452 1 0 gly glycosylation 86:98 arg1 proteins 103:110 proteins 103:110 proteins 103:110 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 5 1 theme proteins 892:899 arg1 hypoglycosylation 864:880 hypoglycosylation 864:880 hypoglycosylation of client proteins 864:899 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 6 2 from flow 951:954 arg1 lipid 1076:1080 the donor lipid 1066:1080 the donor lipid 1066:1080 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 4 3 theme Most 528:531 arg1 strategies 533:542 Most strategies 528:542 Most strategies used for glycoengineering 528:568 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 9 4 theme client 1577:1582 arg1 proteins 1584:1591 client proteins 1577:1591 client proteins 1577:1591 Improving the glycosylation efficiency ensures that not only homogeneous N-glycan structures are generated, but also client proteins are fully glycosylated. 24632452 3 5 theme therapeutic 352:362 arg1 glycoproteins 364:376 therapeutic glycoproteins 352:376 therapeutic glycoproteins 352:376 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 6 6 theme oligosaccharide 1045:1059 arg1 transfer 1029:1036 the transfer 1025:1036 the transfer of the oligosaccharide from the donor lipid onto the protein 1025:1097 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 6 theme oligosaccharide 1045:1059 arg1 flow 951:954 the flow 947:954 the flow of lipid-linked oligosaccharide through the biosynthetic pathway 947:1019 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 7 link lipid-linked 959:970 arg1 oligosaccharide 972:986 lipid-linked oligosaccharide 959:986 lipid-linked oligosaccharide 959:986 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 1 8 link N-linked 77:84 arg1 glycosylation 86:98 N-linked glycosylation 77:98 N-linked glycosylation of proteins 77:110 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 4 9 link lipid-linked 602:613 arg1 biosynthesis 631:642 the lipid-linked oligosaccharide biosynthesis 598:642 the lipid-linked oligosaccharide biosynthesis 598:642 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 5 10 theme lipid-linked 752:763 arg1 biosynthesis 781:792 the lipid-linked oligosaccharide biosynthesis 748:792 the lipid-linked oligosaccharide biosynthesis 748:792 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 3 11 theme glycoengineering 401:416 arg1 work 393:396 pioneering work 382:396 pioneering work of glycoengineering 382:416 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 2 12 theme N-glycan 171:178 arg1 structures 180:189 N-glycan structures 171:189 N-glycan structures 171:189 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 12 theme N-glycan 171:178 arg1 efficiency 211:220 N-glycosylation efficiency 195:220 N-glycosylation efficiency 195:220 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 12 theme N-glycan 171:178 arg1 parameters 234:243 crucial parameters 226:243 crucial parameters in the production of N-glycosylated proteins 226:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 5 13 theme oligosaccharide 765:779 arg1 biosynthesis 781:792 the lipid-linked oligosaccharide biosynthesis 748:792 the lipid-linked oligosaccharide biosynthesis 748:792 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 9 14 theme N-glycan 1533:1540 arg1 structures 1542:1551 not only homogeneous N-glycan structures 1512:1551 not only homogeneous N-glycan structures 1512:1551 Improving the glycosylation efficiency ensures that not only homogeneous N-glycan structures are generated, but also client proteins are fully glycosylated. 24632452 8 15 theme flippase 1450:1457 arg1 overexpression 1418:1431 overexpression 1418:1431 overexpression of an artificial flippase 1418:1457 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 3 16 theme pioneering 382:391 arg1 work 393:396 pioneering work 382:396 pioneering work of glycoengineering 382:416 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 8 17 theme artificial 1439:1448 arg1 flippase 1450:1457 an artificial flippase 1436:1457 an artificial flippase 1436:1457 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 3 18 theme Yeast 291:295 arg1 host 343:346 an attractive production host 318:346 an attractive production host for therapeutic glycoproteins 318:376 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 3 18 theme Yeast 291:295 arg1 cells 297:301 Yeast cells 291:301 Yeast cells 291:301 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 6 19 theme complete 921:928 arg1 N-glycosylation 930:944 complete N-glycosylation 921:944 complete N-glycosylation 921:944 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 1 20 theme N-linked 77:84 arg1 glycosylation 86:98 N-linked glycosylation 77:98 N-linked glycosylation of proteins 77:110 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 9 21 theme glycosylation 1474:1486 arg1 efficiency 1488:1497 the glycosylation efficiency 1470:1497 the glycosylation efficiency 1470:1497 Improving the glycosylation efficiency ensures that not only homogeneous N-glycan structures are generated, but also client proteins are fully glycosylated. 24632452 7 22 theme altered 1237:1243 arg1 specificities 1245:1257 altered specificities 1237:1257 altered specificities for the oligosaccharide and also for the protein acceptor site 1237:1320 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 2 23 from parameters 234:243 arg1 production 252:261 the production 248:261 the production of N-glycosylated proteins 248:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 0 24 from perspective 36:46 arg1 Glycoengineering 0:15 Glycoengineering 0:15 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 0:75 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 24632452 8 25 theme ER 1390:1391 arg1 lumen 1393:1397 the ER lumen 1386:1397 the ER lumen 1386:1397 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 3 26 theme Pichia 435:440 arg1 pastoris 442:449 Pichia pastoris 435:449 Pichia pastoris 435:449 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 4 27 theme oligosaccharide 615:629 arg1 biosynthesis 631:642 the lipid-linked oligosaccharide biosynthesis 598:642 the lipid-linked oligosaccharide biosynthesis 598:642 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 1 28 theme proteins 103:110 arg1 glycosylation 86:98 N-linked glycosylation 77:98 N-linked glycosylation of proteins 77:110 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 8 29 theme lipid-linked 1352:1363 arg1 oligosaccharide 1365:1379 the lipid-linked oligosaccharide 1348:1379 the lipid-linked oligosaccharide 1348:1379 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 3 30 theme capable 476:482 arg1 strains 468:474 yeast strains 462:474 yeast strains capable of producing defined, human-type N-glycans 462:525 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 7 31 theme acceptor 1308:1315 arg1 site 1317:1320 the protein acceptor site 1296:1320 the protein acceptor site 1296:1320 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 4 32 theme lipid-linked 602:613 arg1 biosynthesis 631:642 the lipid-linked oligosaccharide biosynthesis 598:642 the lipid-linked oligosaccharide biosynthesis 598:642 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 7 33 theme oligosaccharyltransferases 1195:1220 arg1 expression 1171:1180 the expression 1167:1180 the expression 1167:1180 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 7 33 theme oligosaccharyltransferases 1195:1220 arg1 tool 1133:1136 A promising tool 1121:1136 A promising tool to improve site occupancy 1121:1162 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 8 34 link lipid-linked 1352:1363 arg1 oligosaccharide 1365:1379 the lipid-linked oligosaccharide 1348:1379 the lipid-linked oligosaccharide 1348:1379 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 5 35 gly hypoglycosylation 864:880 arg1 accumulation 814:825 the accumulation 810:825 the accumulation of intermediate structures 810:852 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 5 35 gly hypoglycosylation 864:880 arg1 proteins 892:899 client proteins 885:899 client proteins 885:899 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 5 36 link lipid-linked 752:763 arg1 biosynthesis 781:792 the lipid-linked oligosaccharide biosynthesis 748:792 the lipid-linked oligosaccharide biosynthesis 748:792 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 0 37 theme yeasts 20:25 arg1 Glycoengineering 0:15 Glycoengineering 0:15 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 0:75 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 24632452 7 38 theme protein 1300:1306 arg1 site 1317:1320 the protein acceptor site 1296:1320 the protein acceptor site 1296:1320 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 8 39 theme oligosaccharide 1365:1379 arg1 flipping 1336:1343 flipping 1336:1343 flipping of the lipid-linked oligosaccharide into the ER lumen 1336:1397 Furthermore, flipping of the lipid-linked oligosaccharide into the ER lumen can be improved by overexpression of an artificial flippase. 24632452 4 40 theme biosynthesis 631:642 arg1 modification 582:593 the modification 578:593 the modification of the lipid-linked oligosaccharide biosynthesis 578:642 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 2 41 theme N-glycosylation 195:209 arg1 structures 180:189 N-glycan structures 171:189 N-glycan structures 171:189 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 41 theme N-glycosylation 195:209 arg1 efficiency 211:220 N-glycosylation efficiency 195:220 N-glycosylation efficiency 195:220 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 41 theme N-glycosylation 195:209 arg1 parameters 234:243 crucial parameters 226:243 crucial parameters in the production of N-glycosylated proteins 226:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 42 theme N-glycosylated 266:279 arg1 proteins 281:288 N-glycosylated proteins 266:288 N-glycosylated proteins 266:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 3 43 theme defined 497:503 arg1 N-glycans 517:525 defined, human-type N-glycans 497:525 defined, human-type N-glycans 497:525 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 2 44 gly N-glycosylated 266:279 arg1 proteins 281:288 N-glycosylated proteins 266:288 N-glycosylated proteins 266:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 9 45 theme homogeneous 1521:1531 arg1 structures 1542:1551 not only homogeneous N-glycan structures 1512:1551 not only homogeneous N-glycan structures 1512:1551 Improving the glycosylation efficiency ensures that not only homogeneous N-glycan structures are generated, but also client proteins are fully glycosylated. 24632452 3 46 theme yeast 462:466 arg1 strains 468:474 yeast strains 462:474 yeast strains capable of producing defined, human-type N-glycans 462:525 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 6 47 from transfer 1029:1036 arg1 lipid 1076:1080 the donor lipid 1066:1080 the donor lipid 1066:1080 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 0 48 theme glycosylation 51:63 arg1 efficiency 65:74 glycosylation efficiency 51:74 glycosylation efficiency 51:74 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 24632452 5 49 theme structures 843:852 arg1 accumulation 814:825 the accumulation 810:825 the accumulation of intermediate structures 810:852 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 3 50 theme production 332:341 arg1 cells 297:301 Yeast cells 291:301 Yeast cells 291:301 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 3 50 theme production 332:341 arg1 host 343:346 an attractive production host 318:346 an attractive production host for therapeutic glycoproteins 318:376 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 5 51 theme client 885:890 arg1 proteins 892:899 client proteins 885:899 client proteins 885:899 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 7 52 theme site 1149:1152 arg1 occupancy 1154:1162 site occupancy 1149:1162 site occupancy 1149:1162 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 7 53 theme protozoan 1185:1193 arg1 oligosaccharyltransferases 1195:1220 protozoan oligosaccharyltransferases 1185:1220 protozoan oligosaccharyltransferases 1185:1220 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 5 54 theme intermediate 830:841 arg1 structures 843:852 intermediate structures 830:852 intermediate structures 830:852 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 9 55 gly glycosylated 1603:1614 arg1 proteins 1584:1591 client proteins 1577:1591 client proteins 1577:1591 Improving the glycosylation efficiency ensures that not only homogeneous N-glycan structures are generated, but also client proteins are fully glycosylated. 24632452 3 56 gly glycoproteins 364:376 arg1 glycoproteins 364:376 therapeutic glycoproteins 352:376 therapeutic glycoproteins 352:376 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 6 57 theme oligosaccharide 972:986 arg1 transfer 1029:1036 the transfer 1025:1036 the transfer of the oligosaccharide from the donor lipid onto the protein 1025:1097 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 57 theme oligosaccharide 972:986 arg1 flow 951:954 the flow 947:954 the flow of lipid-linked oligosaccharide through the biosynthetic pathway 947:1019 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 58 from lipid 1076:1080 arg1 transfer 1029:1036 the transfer 1025:1036 the transfer of the oligosaccharide from the donor lipid onto the protein 1025:1097 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 58 from lipid 1076:1080 arg1 oligosaccharide 1045:1059 the oligosaccharide 1041:1059 the oligosaccharide from the donor lipid 1041:1080 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 58 from lipid 1076:1080 arg1 flow 951:954 the flow 947:954 the flow of lipid-linked oligosaccharide through the biosynthetic pathway 947:1019 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 7 59 contain possess 1229:1235 arg1 expression 1171:1180 the expression 1167:1180 the expression 1167:1180 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 7 59 contain possess 1229:1235 arg2 specificities 1245:1257 altered specificities 1237:1257 altered specificities for the oligosaccharide and also for the protein acceptor site 1237:1320 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 7 59 contain possess 1229:1235 arg1 tool 1133:1136 A promising tool 1121:1136 A promising tool to improve site occupancy 1121:1162 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 6 60 theme donor 1070:1074 arg1 lipid 1076:1080 the donor lipid 1066:1080 the donor lipid 1066:1080 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 6 61 theme lipid-linked 959:970 arg1 oligosaccharide 972:986 lipid-linked oligosaccharide 959:986 lipid-linked oligosaccharide 959:986 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 3 62 dep defined 497:503 arg1 human-type 506:515 human-type 506:515 human-type 506:515 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 5 63 from modifications 731:743 arg1 biosynthesis 781:792 the lipid-linked oligosaccharide biosynthesis 748:792 the lipid-linked oligosaccharide biosynthesis 748:792 However, modifications in the lipid-linked oligosaccharide biosynthesis often result in the accumulation of intermediate structures and cause hypoglycosylation of client proteins. 24632452 3 64 theme attractive 321:330 arg1 cells 297:301 Yeast cells 291:301 Yeast cells 291:301 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 3 64 theme attractive 321:330 arg1 host 343:346 an attractive production host 318:346 an attractive production host for therapeutic glycoproteins 318:376 Yeast cells can be seen as an attractive production host for therapeutic glycoproteins and pioneering work of glycoengineering was performed in Pichia pastoris, realizing yeast strains capable of producing defined, human-type N-glycans. 24632452 2 65 theme crucial 226:232 arg1 structures 180:189 N-glycan structures 171:189 N-glycan structures 171:189 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 65 theme crucial 226:232 arg1 efficiency 211:220 N-glycosylation efficiency 195:220 N-glycosylation efficiency 195:220 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 2 65 theme crucial 226:232 arg1 parameters 234:243 crucial parameters 226:243 crucial parameters in the production of N-glycosylated proteins 226:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 1 66 theme common 131:136 arg1 modifications 156:168 the most common posttranslational modifications 122:168 the most common posttranslational modifications 122:168 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 4 67 theme substrate 670:678 arg1 generation 652:661 the generation 648:661 the generation of the substrate for Golgi-localized glycosyltransferases 648:719 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 24632452 6 68 theme biosynthetic 1000:1011 arg1 pathway 1013:1019 the biosynthetic pathway 996:1019 the biosynthetic pathway 996:1019 In order to ensure complete N-glycosylation, the flow of lipid-linked oligosaccharide through the biosynthetic pathway and the transfer of the oligosaccharide from the donor lipid onto the protein have to be optimized. 24632452 7 69 theme promising 1123:1131 arg1 expression 1171:1180 the expression 1167:1180 the expression 1167:1180 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 7 69 theme promising 1123:1131 arg1 tool 1133:1136 A promising tool 1121:1136 A promising tool to improve site occupancy 1121:1162 A promising tool to improve site occupancy is the expression of protozoan oligosaccharyltransferases, which possess altered specificities for the oligosaccharide and also for the protein acceptor site. 24632452 1 70 theme posttranslational 138:154 arg1 modifications 156:168 the most common posttranslational modifications 122:168 the most common posttranslational modifications 122:168 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 0 71 theme efficiency 65:74 arg1 perspective 36:46 the perspective 32:46 the perspective of glycosylation efficiency 32:74 Glycoengineering of yeasts from the perspective of glycosylation efficiency. 24632452 2 72 theme proteins 281:288 arg1 production 252:261 the production 248:261 the production of N-glycosylated proteins 248:288 N-glycan structures and N-glycosylation efficiency are crucial parameters in the production of N-glycosylated proteins. 24632452 1 73 theme modifications 156:168 arg1 modifications 156:168 the most common posttranslational modifications 122:168 the most common posttranslational modifications 122:168 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 1 73 theme modifications 156:168 arg1 one 115:117 one 115:117 one 115:117 N-linked glycosylation of proteins is one of the most common posttranslational modifications. 24632452 4 74 theme Golgi-localized 684:698 arg1 glycosyltransferases 700:719 Golgi-localized glycosyltransferases 684:719 Golgi-localized glycosyltransferases 684:719 Most strategies used for glycoengineering rely on the modification of the lipid-linked oligosaccharide biosynthesis for the generation of the substrate for Golgi-localized glycosyltransferases. 28620050 4 0 theme N-linked 813:820 arg1 sites 822:826 two N-linked sites 809:826 two N-linked sites 809:826 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 3 1 theme IgG1-Fc 650:656 arg1 function 638:645 function 638:645 function 638:645 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 3 1 theme IgG1-Fc 650:656 arg1 structure 624:632 structure 624:632 structure 624:632 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 6 2 theme N-linked 1193:1200 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 1 3 theme biomimetic 255:264 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 3 theme biomimetic 255:264 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 3 theme biomimetic 255:264 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 8 4 theme tail 1607:1610 arg1 piece 1612:1616 tail piece 1607:1616 tail piece 1607:1616 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 6 5 link N-linked 1193:1200 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 5 6 from Asn-563 1075:1081 arg1 glycosylation 1058:1070 glycosylation 1058:1070 glycosylation at Asn-563 1058:1081 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 2 7 theme cognate 507:513 arg1 receptors 515:523 their cognate receptors 501:523 their cognate receptors with high avidity 501:541 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 3 8 theme critical 731:738 arg1 role 740:743 a similarly critical role 719:743 a similarly critical role 719:743 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 6 9 from introduction 1148:1159 arg1 terminus 1230:1237 the N terminus 1224:1237 the N terminus 1224:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 10 theme sialic 1325:1330 arg1 content 1337:1343 overall sialic acid content 1317:1343 overall sialic acid content of the Fc multimers 1317:1363 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 7 11 theme receptor-binding 1509:1524 arg1 profiles 1526:1533 differential receptor-binding profiles 1496:1533 differential receptor-binding profiles 1496:1533 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 8 12 theme enhanced 1667:1674 arg1 sialylation 1676:1686 enhanced sialylation 1667:1686 enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion 1667:1763 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 3 13 attach attached 597:604 arg3 Asn-297 609:615 Asn-297 609:615 Asn-297 609:615 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 3 13 attach attached 597:604 arg2 glycan 590:595 the unique N-linked glycan 570:595 the unique N-linked glycan attached at Asn-297 570:615 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 5 14 with interactions 1003:1014 arg1 complement 1038:1047 complement 1038:1047 complement 1038:1047 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 5 14 with interactions 1003:1014 arg1 receptors 1024:1032 Fc receptors 1021:1032 Fc receptors 1021:1032 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 4 15 dep Asn-236 884:890 arg1 equivalent 893:902 equivalent 893:902 equivalent 893:902 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 7 16 theme acid 1479:1482 arg1 content 1484:1490 enhanced sialic acid content 1463:1490 enhanced sialic acid content 1463:1490 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 3 17 link N-linked 699:706 arg1 glycan 708:713 the N-linked glycan 695:713 the N-linked glycan 695:713 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 0 18 with interactions 102:113 arg1 receptors 142:150 sialic acid-dependent receptors 120:150 sialic acid-dependent receptors 120:150 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 1 19 theme many 325:328 arg1 diseases 330:337 many diseases 325:337 many diseases 325:337 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 3 20 theme N-linked 581:588 arg1 glycan 590:595 the unique N-linked glycan 570:595 the unique N-linked glycan attached at Asn-297 570:615 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 6 21 gly glycosylation 1202:1214 arg2 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 21 gly glycosylation 1202:1214 arg2 terminus 1230:1237 the N terminus 1224:1237 the N terminus 1224:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 7 22 theme enhanced 1463:1470 arg1 content 1484:1490 enhanced sialic acid content 1463:1490 enhanced sialic acid content 1463:1490 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 2 23 theme multimeric 458:467 arg1 structures 469:478 multimeric structures 458:478 multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity 458:541 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 2 23 theme multimeric 458:467 arg1 hexa-Fcs 481:488 hexa-Fcs 481:488 hexa-Fcs 481:488 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 6 24 theme site 1216:1219 arg1 introduction 1148:1159 introduction 1148:1159 introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) 1148:1293 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 0 25 theme IgG1 61:64 arg1 multimers 66:74 human IgG1 multimers 55:74 human IgG1 multimers 55:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 8 26 link N-linked 1565:1572 arg1 glycans 1574:1580 additional N-linked glycans 1554:1580 additional N-linked glycans 1554:1580 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 3 27 dep unknown 783:789 arg1 has 715:717 has 715:717 has a similarly critical role in multimeric, avidly binding Fcs 715:777 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 1 28 theme fragment 164:171 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 28 theme fragment 164:171 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 28 theme fragment 164:171 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 4 29 theme IgG1 874:877 arg1 Fc 868:869 the Fc 864:869 the Fc of IgG1 864:877 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 5 30 from Asn-297 979:985 arg1 glycosylation 962:974 glycosylation 962:974 glycosylation at Asn-297 962:985 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 6 31 from Asp-221 1268:1274 arg1 Fc 1283:1284 the Fc 1279:1284 the Fc of IgG1 1279:1292 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 32 theme acid 1332:1335 arg1 content 1337:1343 overall sialic acid content 1317:1343 overall sialic acid content of the Fc multimers 1317:1363 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 1 33 theme IVIG 283:286 arg1 therapy 288:294 IVIG therapy 283:294 IVIG therapy 283:294 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 2 34 theme IgG1 420:423 arg1 region 404:409 The Fc region 397:409 The Fc region of human IgG1 (IgG1-Fc) 397:433 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 3 35 contain has 715:717 arg1 glycan 708:713 the N-linked glycan 695:713 the N-linked glycan 695:713 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 3 35 contain has 715:717 arg2 role 740:743 a similarly critical role 719:743 a similarly critical role 719:743 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 3 36 from influence 557:565 arg1 function 638:645 function 638:645 function 638:645 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 3 36 from influence 557:565 arg1 structure 624:632 structure 624:632 structure 624:632 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 6 37 theme overall 1317:1323 arg1 content 1337:1343 overall sialic acid content 1317:1343 overall sialic acid content of the Fc multimers 1317:1363 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 2 38 theme Fc 401:402 arg1 region 404:409 The Fc region 397:409 The Fc region of human IgG1 (IgG1-Fc) 397:433 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 0 39 theme fragment 16:23 arg1 region 45:50 the fragment crystallizable (Fc) region 12:50 the fragment crystallizable (Fc) region of human IgG1 multimers 12:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 7 40 theme IgM 1409:1411 arg1 piece 1418:1422 the IgM tail piece 1405:1422 the IgM tail piece of multimers 1405:1435 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 4 41 theme tail 922:925 arg1 piece 927:931 the tail piece 918:931 the tail piece of IgM 918:938 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 6 42 theme IgG1 1289:1292 arg1 Fc 1283:1284 the Fc 1279:1284 the Fc of IgG1 1279:1292 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 5 43 gly glycosylation 962:974 arg2 Asn-297 979:985 Asn-297 979:985 Asn-297 979:985 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 5 43 gly glycosylation 962:974 arg1 Asn-297 979:985 Asn-297 979:985 Asn-297 979:985 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 2 44 with receptors 515:523 arg1 avidity 535:541 high avidity 530:541 high avidity 530:541 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 1 45 dep always 379:384 arg1 not 375:377 not 375:377 not 375:377 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 6 46 theme glycosylation 1202:1214 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 47 theme Fc 1352:1353 arg1 multimers 1355:1363 the Fc multimers 1348:1363 the Fc multimers 1348:1363 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 3 48 dep structure 624:632 arg1 the 620:622 the 620:622 the 620:622 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 6 49 theme occupied 1184:1191 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 50 theme additional 1167:1176 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 1 51 theme crystallizable 173:186 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 51 theme crystallizable 173:186 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 51 theme crystallizable 173:186 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 4 52 dep Asn-77 831:836 arg1 equivalent 839:848 equivalent 839:848 equivalent 839:848 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 1 53 theme Fc 189:190 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 53 theme Fc 189:190 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 53 theme Fc 189:190 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 4 54 link N-linked 813:820 arg1 sites 822:826 two N-linked sites 809:826 two N-linked sites 809:826 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 8 55 theme monomers 1621:1628 arg1 piece 1612:1616 tail piece 1607:1616 tail piece 1607:1616 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 8 55 theme monomers 1621:1628 arg1 hinge 1598:1602 the hinge 1594:1602 the hinge 1594:1602 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 3 56 theme N-linked 699:706 arg1 glycan 708:713 the N-linked glycan 695:713 the N-linked glycan 695:713 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 0 57 theme acid-dependent 127:140 arg1 receptors 142:150 sialic acid-dependent receptors 120:150 sialic acid-dependent receptors 120:150 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 3 58 dep multimeric 748:757 arg1 binding 767:773 binding 767:773 binding 767:773 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 8 59 theme multimers 1633:1641 arg1 piece 1612:1616 tail piece 1607:1616 tail piece 1607:1616 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 8 59 theme multimers 1633:1641 arg1 hinge 1598:1602 the hinge 1594:1602 the hinge 1594:1602 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 1 60 theme Fc-fusion 205:213 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 60 theme Fc-fusion 205:213 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 60 theme Fc-fusion 205:213 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 7 61 theme differential 1496:1507 arg1 profiles 1526:1533 differential receptor-binding profiles 1496:1533 differential receptor-binding profiles 1496:1533 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 8 62 with molecules 1652:1660 arg1 sialylation 1676:1686 enhanced sialylation 1667:1686 enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion 1667:1763 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 8 63 gly sialylation 1676:1686 arg1 molecules 1652:1660 molecules 1652:1660 molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion 1652:1763 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 6 64 theme N 1228:1228 arg1 terminus 1230:1237 the N terminus 1224:1237 the N terminus 1224:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 4 65 from Asn-297 853:859 arg1 Fc 868:869 the Fc 864:869 the Fc of IgG1 864:877 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 3 66 theme critical 548:555 arg1 influence 557:565 The critical influence 544:565 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc 544:656 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 7 67 from replacement 1379:1389 arg1 piece 1418:1422 the IgM tail piece 1405:1422 the IgM tail piece of multimers 1405:1435 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 3 68 theme glycan 590:595 arg1 influence 557:565 The critical influence 544:565 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc 544:656 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 7 69 with monomers 1449:1456 arg1 content 1484:1490 enhanced sialic acid content 1463:1490 enhanced sialic acid content 1463:1490 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 7 69 with monomers 1449:1456 arg1 profiles 1526:1533 differential receptor-binding profiles 1496:1533 differential receptor-binding profiles 1496:1533 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 4 70 contain contains 800:807 arg1 Hexa-Fc 792:798 Hexa-Fc 792:798 Hexa-Fc 792:798 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 4 70 contain contains 800:807 arg1 Asn-77 831:836 Asn-77 831:836 Asn-77 (equivalent to Asn-297 in the Fc of IgG1) 831:878 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 4 70 contain contains 800:807 arg2 sites 822:826 two N-linked sites 809:826 two N-linked sites 809:826 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 4 70 contain contains 800:807 arg1 Asn-236 884:890 Asn-236 884:890 Asn-236 (equivalent to Asn-563 in the tail piece of IgM) 884:939 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 4 70 contain contains 800:807 arg2 Hexa-Fc 792:798 Hexa-Fc 792:798 Hexa-Fc 792:798 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 7 71 theme sialic 1472:1477 arg1 content 1484:1490 enhanced sialic acid content 1463:1490 enhanced sialic acid content 1463:1490 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 2 72 theme high 530:533 arg1 avidity 535:541 high avidity 530:541 high avidity 530:541 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 0 73 theme human 55:59 arg1 multimers 66:74 human IgG1 multimers 55:74 human IgG1 multimers 55:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 3 74 theme unique 574:579 arg1 glycan 590:595 the unique N-linked glycan 570:595 the unique N-linked glycan attached at Asn-297 570:615 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 5 75 gly glycosylation 1058:1070 arg1 Asn-563 1075:1081 Asn-563 1075:1081 Asn-563 1075:1081 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 5 75 gly glycosylation 1058:1070 arg2 Asn-563 1075:1081 Asn-563 1075:1081 Asn-563 1075:1081 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 3 76 theme multimeric 748:757 arg1 Fcs 775:777 multimeric, avidly binding Fcs 748:777 multimeric, avidly binding Fcs 748:777 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 0 77 theme multimers 66:74 arg1 monomers 80:87 monomers 80:87 monomers to fine-tune interactions with sialic acid-dependent receptors 80:150 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 0 77 theme multimers 66:74 arg1 multimers 66:74 human IgG1 multimers 55:74 human IgG1 multimers 55:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 0 77 theme multimers 66:74 arg1 region 45:50 the fragment crystallizable (Fc) region 12:50 the fragment crystallizable (Fc) region of human IgG1 multimers 12:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 5 78 theme Fc 1021:1022 arg1 receptors 1024:1032 Fc receptors 1021:1032 Fc receptors 1021:1032 We report here that glycosylation at Asn-297 is critical for interactions with Fc receptors and complement and that glycosylation at Asn-563 is essential for controlling multimerization. 28620050 1 79 theme Multimeric 153:162 arg1 replacements 266:277 biomimetic replacements 255:277 biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient 255:394 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 79 theme Multimeric 153:162 arg1 regions 193:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions 153:199 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 1 79 theme Multimeric 153:162 arg1 proteins 215:222 Fc-fusion proteins 205:222 Fc-fusion proteins 205:222 Multimeric fragment crystallizable (Fc) regions and Fc-fusion proteins are actively being explored as biomimetic replacements for IVIG therapy, which is deployed to manage many diseases and conditions but is expensive and not always efficient. 28620050 7 80 theme multimers 1427:1435 arg1 piece 1418:1422 the IgM tail piece 1405:1422 the IgM tail piece of multimers 1405:1435 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 0 81 theme fine-tune 92:100 arg1 interactions 102:113 fine-tune interactions 92:113 fine-tune interactions with sialic acid-dependent receptors 92:150 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 6 82 theme equivalent 1254:1263 arg1 position 1242:1249 position 1 1242:1251 position 1 (equivalent to Asp-221 in the Fc of IgG1) 1242:1293 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 3 83 link N-linked 581:588 arg1 glycan 590:595 the unique N-linked glycan 570:595 the unique N-linked glycan attached at Asn-297 570:615 The critical influence of the unique N-linked glycan attached at Asn-297 on the structure and function of IgG1-Fc is well documented; however, whether the N-linked glycan has a similarly critical role in multimeric, avidly binding Fcs, is unknown. 28620050 0 84 theme crystallizable 25:38 arg1 region 45:50 the fragment crystallizable (Fc) region 12:50 the fragment crystallizable (Fc) region of human IgG1 multimers 12:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 6 85 from terminus 1230:1237 arg1 introduction 1148:1159 introduction 1148:1159 introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) 1148:1293 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 6 85 from terminus 1230:1237 arg1 site 1216:1219 an additional fully occupied N-linked glycosylation site 1164:1219 an additional fully occupied N-linked glycosylation site at the N terminus 1164:1237 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 8 86 theme pathogen 1747:1754 arg1 invasion 1756:1763 pathogen invasion 1747:1763 pathogen invasion 1747:1763 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 2 87 theme human 414:418 arg1 IgG1-Fc 426:432 IgG1-Fc 426:432 IgG1-Fc 426:432 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 2 87 theme human 414:418 arg1 IgG1 420:423 human IgG1 414:423 human IgG1 (IgG1-Fc) 414:433 The Fc region of human IgG1 (IgG1-Fc) can be engineered into multimeric structures (hexa-Fcs) that bind their cognate receptors with high avidity. 28620050 0 88 theme Fc 41:42 arg1 region 45:50 the fragment crystallizable (Fc) region 12:50 the fragment crystallizable (Fc) region of human IgG1 multimers 12:74 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 6 89 from position 1242:1249 arg1 introduction 1148:1159 introduction 1148:1159 introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) 1148:1293 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 28620050 8 90 theme N-linked 1565:1572 arg1 glycans 1574:1580 additional N-linked glycans 1554:1580 additional N-linked glycans 1554:1580 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 7 91 theme tail 1413:1416 arg1 piece 1418:1422 the IgM tail piece 1405:1422 the IgM tail piece of multimers 1405:1435 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 0 92 theme sialic 120:125 arg1 receptors 142:150 sialic acid-dependent receptors 120:150 sialic acid-dependent receptors 120:150 Engineering the fragment crystallizable (Fc) region of human IgG1 multimers and monomers to fine-tune interactions with sialic acid-dependent receptors. 28620050 8 93 theme additional 1554:1563 arg1 glycans 1574:1580 additional N-linked glycans 1554:1580 additional N-linked glycans 1554:1580 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 4 94 from Asn-563 907:913 arg1 piece 927:931 the tail piece 918:931 the tail piece of IgM 918:938 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 4 95 theme IgM 936:938 arg1 piece 927:931 the tail piece 918:931 the tail piece of IgM 918:938 Hexa-Fc contains two N-linked sites at Asn-77 (equivalent to Asn-297 in the Fc of IgG1) and Asn-236 (equivalent to Asn-563 in the tail piece of IgM). 28620050 8 96 theme glycans 1574:1580 arg1 insertion 1541:1549 insertion 1541:1549 insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers 1541:1641 Thus insertion of additional N-linked glycans into either the hinge or tail piece of monomers or multimers leads to molecules with enhanced sialylation that may be suitable for managing inflammation or blocking pathogen invasion. 28620050 7 97 theme Cys-575 1394:1400 arg1 replacement 1379:1389 replacement 1379:1389 replacement of Cys-575 in the IgM tail piece of multimers 1379:1435 Furthermore, replacement of Cys-575 in the IgM tail piece of multimers resulted in monomers with enhanced sialic acid content and differential receptor-binding profiles. 28620050 6 98 theme multimers 1355:1363 arg1 content 1337:1343 overall sialic acid content 1317:1343 overall sialic acid content of the Fc multimers 1317:1363 We also found that introduction of an additional fully occupied N-linked glycosylation site at the N terminus at position 1 (equivalent to Asp-221 in the Fc of IgG1) dramatically enhances overall sialic acid content of the Fc multimers. 26512079 9 0 from glycans 1861:1867 arg1 protein 1891:1897 the HIV-1 envelope protein 1872:1897 the HIV-1 envelope protein 1872:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 1 1 from infection 243:251 arg1 part 256:259 part 256:259 part 256:259 HIV-1 establishes persistent infection in part due to its ability to evade host immune responses. 26512079 3 2 theme mutant 649:654 arg1 virus 656:660 the mutant virus 645:660 the mutant virus 645:660 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 2 3 theme masking 348:354 arg1 sites 366:370 masking conserved sites 348:370 masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs) 348:436 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 2 3 theme masking 348:354 arg1 targets 381:387 targets 381:387 targets for some broadly neutralizing antibodies (bNAbs) 381:436 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 4 4 from glycans 863:869 arg1 regions 888:894 the conserved regions 874:894 the conserved regions of HIV-1 Env 874:907 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 4 5 theme N7 798:799 arg1 glycan 801:806 the N7 glycan 794:806 the N7 glycan 794:806 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 5 6 from origins 1123:1129 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 6 from origins 1123:1129 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 11 7 theme N-linked 2177:2184 arg1 glycan 2186:2191 a single N-linked glycan 2168:2191 a single N-linked glycan 2168:2191 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 3 8 theme site 531:534 arg1 removal 468:474 removal 468:474 removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 468:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 5 9 theme HIV-1 1071:1075 arg1 strains 1077:1083 HIV-1 strains 1071:1083 HIV-1 strains representing different clades 1071:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 12 10 theme coreceptor 2535:2544 arg1 sites 2554:2558 the receptor and coreceptor binding sites 2518:2558 sites 2554:2558 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 3 11 theme binding 669:675 arg1 site 677:680 CD4 binding site 665:680 CD4 binding site 665:680 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 0 12 theme Binding 199:205 arg1 Sites 207:211 the Receptor and Coreceptor Binding Sites 171:211 Sites 207:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 3 13 from removal 468:474 arg1 N7 563:564 N7 563:564 N7 563:564 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 13 from removal 468:474 arg1 gp120 590:594 the surface antigen gp120 570:594 the surface antigen gp120 of HIV-1 570:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 13 from removal 468:474 arg1 residue 550:556 amino acid residue 197 539:560 amino acid residue 197 (N7) 539:565 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 14 theme PNLG 525:528 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 6 15 theme conserved 1396:1404 arg1 epitopes 1406:1413 these conserved epitopes 1390:1413 these conserved epitopes 1390:1413 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 11 16 theme unique 2201:2206 arg1 role 2222:2225 a unique and conserved role 2199:2225 a unique and conserved role among conserved glycans on HIV-1 gp120 2199:2264 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 8 17 theme N7 1593:1594 arg1 glycan 1596:1601 the N7 glycan 1589:1601 the N7 glycan 1589:1601 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 9 18 link N-linked 1852:1859 arg1 glycans 1861:1867 N-linked glycans 1852:1867 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein 1840:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 4 19 theme HIV-1 835:839 arg1 isolates 841:848 diverse HIV-1 isolates 827:848 diverse HIV-1 isolates 827:848 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 3 20 theme amino 539:543 arg1 N7 563:564 N7 563:564 N7 563:564 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 20 theme amino 539:543 arg1 residue 550:556 amino acid residue 197 539:560 amino acid residue 197 (N7) 539:565 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 6 21 theme conserved 1367:1375 arg1 role 1377:1380 a conserved role 1365:1380 a conserved role masking these conserved epitopes 1365:1413 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 3 22 link N-linked 508:515 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 23 theme wild-type 726:734 arg1 counterpart 741:751 its wild-type (WT) counterpart 722:751 its wild-type (WT) counterpart 722:751 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 8 24 theme important 1612:1620 arg1 role 1636:1639 an important and conserved role 1609:1639 an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes 1609:1710 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 6 25 theme V3 1300:1301 arg1 antibodies 1317:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 0 26 theme Broadly 131:137 arg1 Antibodies 152:161 Broadly Neutralizing Antibodies 131:161 Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 131:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 8 27 theme conserved 1626:1634 arg1 role 1636:1639 an important and conserved role 1609:1639 an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes 1609:1710 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 7 28 theme virus 1456:1460 arg1 sensitivity 1462:1472 virus sensitivity 1456:1472 virus sensitivity to neutralizing antibodies directed against the V2 loop epitope 1456:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 11 29 theme Env 2393:2395 arg1 integrity 2376:2384 the integrity 2372:2384 the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4 2372:2474 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 10 30 theme specific 1999:2006 arg1 glycans 2008:2014 specific glycans 1999:2014 specific glycans 1999:2014 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 11 31 theme HIV-1 2254:2258 arg1 gp120 2260:2264 HIV-1 gp120 2254:2264 HIV-1 gp120 2254:2264 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 3 32 theme antigen 582:588 arg1 gp120 590:594 the surface antigen gp120 570:594 the surface antigen gp120 of HIV-1 570:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 7 33 theme glycan 1446:1451 arg1 effect 1429:1434 the effect 1425:1434 the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope 1425:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 7 33 theme glycan 1446:1451 arg1 dependent 1549:1557 dependent 1549:1557 dependent 1549:1557 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 4 34 theme conserved 878:886 arg1 regions 888:894 the conserved regions 874:894 the conserved regions of HIV-1 Env 874:907 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 3 35 from residue 550:556 arg1 removal 468:474 removal 468:474 removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 468:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 35 from residue 550:556 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 0 36 theme Surface 44:50 arg1 Antigen 52:58 the Surface Antigen 40:58 the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 40:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 9 37 theme N-linked 1852:1859 arg1 glycans 1861:1867 N-linked glycans 1852:1867 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein 1840:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 5 38 theme tissue 1116:1121 arg1 origins 1123:1129 tissue origins 1116:1129 tissue origins 1116:1129 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 39 theme Env 1013:1015 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 4 40 theme other 857:861 arg1 glycans 863:869 other glycans 857:869 other glycans in the conserved regions of HIV-1 Env 857:907 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 11 41 theme conserved 2212:2220 arg1 role 2222:2225 a unique and conserved role 2199:2225 a unique and conserved role among conserved glycans on HIV-1 gp120 2199:2264 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 3 42 from site 531:534 arg1 gp120 590:594 the surface antigen gp120 570:594 the surface antigen gp120 of HIV-1 570:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 0 43 theme Immunodeficiency 69:84 arg1 Type 92:95 Human Immunodeficiency Virus Type 1 63:97 Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 63:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 11 44 theme binding 2341:2347 arg1 site 2349:2352 the receptor and coreceptor binding site 2313:2352 site 2349:2352 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 9 45 theme HIV-1 1876:1880 arg1 protein 1891:1897 the HIV-1 envelope protein 1872:1897 the HIV-1 envelope protein 1872:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 0 46 theme Type 92:95 arg1 Antigen 52:58 the Surface Antigen 40:58 the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 40:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 3 47 dep wild-type 726:734 arg1 WT 737:738 WT 737:738 WT 737:738 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 12 48 theme single 2582:2587 arg1 glycan 2589:2594 a single glycan 2580:2594 a single glycan 2580:2594 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 12 49 theme select 2611:2616 arg1 modification 2625:2636 select glycan modification 2611:2636 select glycan modification 2611:2636 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 11 50 theme mutant 2448:2453 arg1 gp120 2455:2459 the mutant gp120 2444:2459 the mutant gp120 2444:2459 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 0 51 theme Conserved 0:8 arg1 Role 10:13 Conserved Role 0:13 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 0:212 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 8 52 theme epitopes 1703:1710 arg1 stability 1667:1675 stability 1667:1675 stability 1667:1675 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 8 52 theme epitopes 1703:1710 arg1 accessibility 1681:1693 accessibility 1681:1693 accessibility 1681:1693 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 8 52 theme epitopes 1703:1710 arg1 structure 1656:1664 structure 1656:1664 structure 1656:1664 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 7 53 theme loop 1525:1528 arg1 epitope 1530:1536 the V2 loop epitope 1518:1536 the V2 loop epitope 1518:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 6 54 theme CD4bs- 1289:1294 arg1 antibodies 1317:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 5 55 theme glycan 1050:1055 arg1 origins 1123:1129 tissue origins 1116:1129 tissue origins 1116:1129 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 55 theme glycan 1050:1055 arg1 role 966:969 the role 962:969 the role of PNLGs in the conserved region of HIV-1 Env 962:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 55 theme glycan 1050:1055 arg1 role 1035:1038 particularly the role 1018:1038 particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades 1018:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 55 theme glycan 1050:1055 arg1 sensitivities 1170:1182 neutralization sensitivities 1155:1182 neutralization sensitivities 1155:1182 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 55 theme glycan 1050:1055 arg1 usages 1143:1148 coreceptor usages 1132:1148 coreceptor usages 1132:1148 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 4 56 theme Env 905:907 arg1 regions 888:894 the conserved regions 874:894 the conserved regions of HIV-1 Env 874:907 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 11 57 theme conserved 2233:2241 arg1 glycans 2243:2249 conserved glycans 2233:2249 conserved glycans 2233:2249 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 11 58 link N-linked 2177:2184 arg1 glycan 2186:2191 a single N-linked glycan 2168:2191 a single N-linked glycan 2168:2191 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 2 59 theme neutralizing 406:417 arg1 bNAbs 431:435 bNAbs 431:435 bNAbs 431:435 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 2 59 theme neutralizing 406:417 arg1 antibodies 419:428 some broadly neutralizing antibodies 393:428 some broadly neutralizing antibodies (bNAbs) 393:436 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 6 60 theme isolates 1277:1284 arg1 sensitivity 1248:1258 the sensitivity 1244:1258 the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies 1244:1326 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 1 61 theme immune 294:299 arg1 responses 301:309 host immune responses 289:309 host immune responses 289:309 HIV-1 establishes persistent infection in part due to its ability to evade host immune responses. 26512079 0 62 theme N-Linked 21:28 arg1 Glycan 30:35 an N-Linked Glycan 18:35 an N-Linked Glycan 18:35 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 4 63 theme similar 917:923 arg1 functions 925:933 similar functions 917:933 similar functions 917:933 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 11 64 from role 2222:2225 arg1 gp120 2260:2264 HIV-1 gp120 2254:2264 HIV-1 gp120 2254:2264 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 6 65 theme diverse 1263:1269 arg1 isolates 1277:1284 diverse HIV-1 isolates 1263:1284 diverse HIV-1 isolates 1263:1284 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 8 66 theme coreceptor 1729:1738 arg1 region 1748:1753 the CD4bs and coreceptor binding region 1715:1753 region 1748:1753 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 8 67 dep structure 1656:1664 arg1 the 1652:1654 the 1652:1654 the 1652:1654 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 5 68 theme strains 1077:1083 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 9 69 theme host 1955:1958 arg1 responses 1967:1975 host immune responses 1955:1975 host immune responses 1955:1975 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 3 70 theme N-linked 508:515 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 5 71 theme different 1098:1106 arg1 clades 1108:1113 different clades 1098:1113 different clades 1098:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 3 72 theme CD4 665:667 arg1 site 677:680 CD4 binding site 665:680 CD4 binding site 665:680 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 5 73 from role 1035:1038 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 73 from role 1035:1038 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 2 74 theme conserved 356:364 arg1 sites 366:370 masking conserved sites 348:370 masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs) 348:436 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 2 74 theme conserved 356:364 arg1 targets 381:387 targets 381:387 targets for some broadly neutralizing antibodies (bNAbs) 381:436 Occlusion by glycans contributes to masking conserved sites that are targets for some broadly neutralizing antibodies (bNAbs). 26512079 5 75 theme HIV-1 1007:1011 arg1 Env 1013:1015 HIV-1 Env 1007:1015 HIV-1 Env 1007:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 12 76 theme sites 2554:2558 arg1 antigenicity 2502:2513 the antigenicity 2498:2513 the antigenicity of the receptor and coreceptor binding sites 2498:2558 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 11 77 theme single 2170:2175 arg1 glycan 2186:2191 a single N-linked glycan 2168:2191 a single N-linked glycan 2168:2191 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 3 78 from gp120 590:594 arg1 removal 468:474 removal 468:474 removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 468:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 79 theme virus 656:660 arg1 sensitivity 630:640 neutralization sensitivity 615:640 neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies 615:708 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 0 80 theme Coreceptor 188:197 arg1 Sites 207:211 the Receptor and Coreceptor Binding Sites 171:211 Sites 207:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 10 81 theme conserved 2023:2031 arg1 regions 2033:2039 the conserved regions 2019:2039 the conserved regions of HIV-1 Env 2019:2052 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 8 82 theme binding 1740:1746 arg1 region 1748:1753 the CD4bs and coreceptor binding region 1715:1753 region 1748:1753 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 9 83 from responses 1967:1975 arg1 escape 1943:1948 viral escape 1937:1948 viral escape from host immune responses 1937:1975 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 3 84 theme acid 545:548 arg1 N7 563:564 N7 563:564 N7 563:564 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 84 theme acid 545:548 arg1 residue 550:556 amino acid residue 197 539:560 amino acid residue 197 (N7) 539:565 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 7 85 from effect 1429:1434 arg1 sensitivity 1462:1472 virus sensitivity 1456:1472 virus sensitivity to neutralizing antibodies directed against the V2 loop epitope 1456:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 12 86 theme binding 2546:2552 arg1 sites 2554:2558 the receptor and coreceptor binding sites 2518:2558 sites 2554:2558 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 0 87 theme Virus 110:114 arg1 Sensitivity 116:126 Virus Sensitivity 110:126 Virus Sensitivity 110:126 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 10 88 theme Env 2050:2052 arg1 regions 2033:2039 the conserved regions 2019:2039 the conserved regions of HIV-1 Env 2019:2052 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 7 89 theme neutralizing 1477:1488 arg1 antibodies 1490:1499 neutralizing antibodies 1477:1499 neutralizing antibodies directed against the V2 loop epitope 1477:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 4 90 theme diverse 827:833 arg1 isolates 841:848 diverse HIV-1 isolates 827:848 diverse HIV-1 isolates 827:848 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 5 91 from usages 1143:1148 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 91 from usages 1143:1148 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 3 92 theme conserved 488:496 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 3 93 theme -directed 689:697 arg1 antibodies 699:708 CD4 binding site (CD4bs)-directed antibodies 665:708 CD4 binding site (CD4bs)-directed antibodies 665:708 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 8 94 theme potential 1776:1784 arg1 target 1786:1791 a potential target 1774:1791 a potential target for the design of immunogens and therapeutics 1774:1837 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 9 95 dep IMPORTANCE 1840:1849 arg1 glycans 1861:1867 N-linked glycans 1852:1867 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein 1840:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 11 96 theme site 2349:2352 arg1 exposure 2284:2291 the exposure 2280:2291 the exposure 2280:2291 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 11 96 theme site 2349:2352 arg1 stability 2300:2308 the stability 2296:2308 the stability of the receptor and coreceptor binding site 2296:2352 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 6 97 theme glycan 1227:1232 arg1 absence 1209:1215 the absence 1205:1215 the absence of the N7 glycan 1205:1232 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 0 98 theme Neutralizing 139:150 arg1 Antibodies 152:161 Broadly Neutralizing Antibodies 131:161 Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 131:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 5 99 theme PNLGs 974:978 arg1 origins 1123:1129 tissue origins 1116:1129 tissue origins 1116:1129 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 99 theme PNLGs 974:978 arg1 role 966:969 the role 962:969 the role of PNLGs in the conserved region of HIV-1 Env 962:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 99 theme PNLGs 974:978 arg1 role 1035:1038 particularly the role 1018:1038 particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades 1018:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 99 theme PNLGs 974:978 arg1 sensitivities 1170:1182 neutralization sensitivities 1155:1182 neutralization sensitivities 1155:1182 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 99 theme PNLGs 974:978 arg1 usages 1143:1148 coreceptor usages 1132:1148 coreceptor usages 1132:1148 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 4 100 theme glycan 801:806 arg1 role 786:789 the role 782:789 the role of the N7 glycan 782:806 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 5 101 from sensitivities 1170:1182 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 101 from sensitivities 1170:1182 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 10 102 theme glycans 2008:2014 arg1 role 1991:1994 the role 1987:1994 the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies 1987:2121 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 5 103 theme coreceptor 1132:1141 arg1 usages 1143:1148 coreceptor usages 1132:1148 coreceptor usages 1132:1148 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 3 104 theme surface 574:580 arg1 gp120 590:594 the surface antigen gp120 570:594 the surface antigen gp120 of HIV-1 570:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 7 105 theme N7 1443:1444 arg1 glycan 1446:1451 the N7 glycan 1439:1451 the N7 glycan 1439:1451 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 10 106 theme HIV-1 2044:2048 arg1 Env 2050:2052 HIV-1 Env 2044:2052 HIV-1 Env 2044:2052 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 8 107 theme immunogens 1811:1820 arg1 design 1801:1806 the design 1797:1806 the design of immunogens and therapeutics 1797:1837 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 6 108 theme N7 1224:1225 arg1 glycan 1227:1232 the N7 glycan 1220:1232 the N7 glycan 1220:1232 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 0 109 theme Human 63:67 arg1 Type 92:95 Human Immunodeficiency Virus Type 1 63:97 Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 63:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 3 110 dep -directed 689:697 arg1 site 677:680 CD4 binding site 665:680 CD4 binding site 665:680 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 0 111 from Role 10:13 arg1 Antigen 52:58 the Surface Antigen 40:58 the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 40:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 10 112 theme epitope 2068:2074 arg1 recognition 2076:2086 epitope recognition 2068:2086 epitope recognition 2068:2086 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 0 113 theme Virus 86:90 arg1 Type 92:95 Human Immunodeficiency Virus Type 1 63:97 Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites 63:211 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 3 114 dep removal 468:474 arg1 increases 605:613 increases 605:613 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart 605:751 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 11 115 theme coreceptor 2330:2339 arg1 site 2349:2352 the receptor and coreceptor binding site 2313:2352 site 2349:2352 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 12 116 theme potential 2647:2655 arg1 strategy 2657:2664 a potential strategy 2645:2664 a potential strategy for the design of HIV-1 vaccine candidates 2645:2707 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 11 117 theme receptor 2317:2324 arg1 exposure 2284:2291 the exposure 2280:2291 the exposure 2280:2291 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 11 117 theme receptor 2317:2324 arg1 stability 2300:2308 the stability 2296:2308 the stability of the receptor and coreceptor binding site 2296:2352 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 9 118 theme envelope 1882:1889 arg1 protein 1891:1897 the HIV-1 envelope protein 1872:1897 the HIV-1 envelope protein 1872:1897 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 3 119 theme Previous 439:446 arg1 work 448:451 Previous work 439:451 Previous work 439:451 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 6 120 theme loop-directed 1303:1315 arg1 antibodies 1317:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 CD4bs- and V3 loop-directed antibodies 1289:1326 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 12 121 theme receptor 2522:2529 arg1 antigenicity 2502:2513 the antigenicity 2498:2513 the antigenicity of the receptor and coreceptor binding sites 2498:2558 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 11 122 theme gp120 2455:2459 arg1 infection 2416:2424 viral infection 2410:2424 viral infection 2410:2424 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 11 122 theme gp120 2455:2459 arg1 ability 2433:2439 the ability 2429:2439 the ability of the mutant gp120 to bind to CD4 2429:2474 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 12 123 theme candidates 2698:2707 arg1 design 2674:2679 the design 2670:2679 the design of HIV-1 vaccine candidates 2670:2707 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 5 124 theme neutralization 1155:1168 arg1 sensitivities 1170:1182 neutralization sensitivities 1155:1182 neutralization sensitivities 1155:1182 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 125 theme N7 1047:1048 arg1 glycan 1050:1055 the N7 glycan 1043:1055 the N7 glycan 1043:1055 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 8 126 theme bNAb 1698:1701 arg1 epitopes 1703:1710 bNAb epitopes 1698:1710 bNAb epitopes 1698:1710 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 12 127 theme glycan 2618:2623 arg1 modification 2625:2636 select glycan modification 2611:2636 select glycan modification 2611:2636 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 3 128 theme HIV-1 599:603 arg1 gp120 590:594 the surface antigen gp120 570:594 the surface antigen gp120 of HIV-1 570:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 12 129 theme vaccine 2690:2696 arg1 candidates 2698:2707 HIV-1 vaccine candidates 2684:2707 HIV-1 vaccine candidates 2684:2707 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 0 130 theme Glycan 30:35 arg1 Role 10:13 Conserved Role 0:13 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 0:212 Conserved Role of an N-Linked Glycan on the Surface Antigen of Human Immunodeficiency Virus Type 1 Modulating Virus Sensitivity to Broadly Neutralizing Antibodies against the Receptor and Coreceptor Binding Sites. 26512079 1 131 theme host 289:292 arg1 responses 301:309 host immune responses 289:309 host immune responses 289:309 HIV-1 establishes persistent infection in part due to its ability to evade host immune responses. 26512079 7 132 theme V2 1522:1523 arg1 epitope 1530:1536 the V2 loop epitope 1518:1536 the V2 loop epitope 1518:1536 However, the effect of the N7 glycan on virus sensitivity to neutralizing antibodies directed against the V2 loop epitope is isolate dependent. 26512079 4 133 theme HIV-1 899:903 arg1 Env 905:907 HIV-1 Env 899:907 HIV-1 Env 899:907 However, it is not clear if the role of the N7 glycan is conserved among diverse HIV-1 isolates and if other glycans in the conserved regions of HIV-1 Env display similar functions. 26512079 8 134 theme therapeutics 1826:1837 arg1 design 1801:1806 the design 1797:1806 the design of immunogens and therapeutics 1797:1837 These findings indicate that the N7 glycan plays an important and conserved role modulating the structure, stability, or accessibility of bNAb epitopes in the CD4bs and coreceptor binding region, thus representing a potential target for the design of immunogens and therapeutics. 26512079 5 135 from role 966:969 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 5 135 from role 966:969 arg1 panel 1062:1066 a panel 1060:1066 a panel of HIV-1 strains representing different clades 1060:1113 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 6 136 theme HIV-1 1271:1275 arg1 isolates 1277:1284 diverse HIV-1 isolates 1263:1284 diverse HIV-1 isolates 1263:1284 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 5 137 theme conserved 987:995 arg1 region 997:1002 the conserved region 983:1002 the conserved region of HIV-1 Env 983:1015 In this work, we examined the role of PNLGs in the conserved region of HIV-1 Env, particularly the role of the N7 glycan in a panel of HIV-1 strains representing different clades, tissue origins, coreceptor usages, and neutralization sensitivities. 26512079 9 138 theme viral 1937:1941 arg1 escape 1943:1948 viral escape 1937:1948 viral escape from host immune responses 1937:1975 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 3 139 theme potential 498:506 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 10 140 theme neutralizing 2099:2110 arg1 antibodies 2112:2121 broadly neutralizing antibodies 2091:2121 broadly neutralizing antibodies 2091:2121 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 11 141 theme viral 2410:2414 arg1 infection 2416:2424 viral infection 2410:2424 viral infection 2410:2424 We show here that a single N-linked glycan plays a unique and conserved role among conserved glycans on HIV-1 gp120 in modulating the exposure or the stability of the receptor and coreceptor binding site without affecting the integrity of the Env in mediating viral infection or the ability of the mutant gp120 to bind to CD4. 26512079 3 142 theme neutralization 615:628 arg1 sensitivity 630:640 neutralization sensitivity 615:640 neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies 615:708 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 26512079 12 143 theme HIV-1 2684:2688 arg1 candidates 2698:2707 HIV-1 vaccine candidates 2684:2707 HIV-1 vaccine candidates 2684:2707 The observation that the antigenicity of the receptor and coreceptor binding sites can be modulated by a single glycan indicates that select glycan modification offers a potential strategy for the design of HIV-1 vaccine candidates. 26512079 6 144 theme N7 1349:1350 arg1 glycan 1352:1357 the N7 glycan 1345:1357 the N7 glycan 1345:1357 We demonstrate that the absence of the N7 glycan increases the sensitivity of diverse HIV-1 isolates to CD4bs- and V3 loop-directed antibodies, indicating that the N7 glycan plays a conserved role masking these conserved epitopes. 26512079 1 145 theme persistent 232:241 arg1 infection 243:251 persistent infection 232:251 persistent infection in part 232:259 HIV-1 establishes persistent infection in part due to its ability to evade host immune responses. 26512079 9 146 theme immune 1960:1965 arg1 responses 1967:1975 host immune responses 1955:1975 host immune responses 1955:1975 IMPORTANCE: N-linked glycans on the HIV-1 envelope protein have been postulated to contribute to viral escape from host immune responses. 26512079 10 147 from role 1991:1994 arg1 regions 2033:2039 the conserved regions 2019:2039 the conserved regions of HIV-1 Env 2019:2052 However, the role of specific glycans in the conserved regions of HIV-1 Env in modulating epitope recognition by broadly neutralizing antibodies has not been well defined. 26512079 3 148 theme glycan 517:522 arg1 site 531:534 a highly conserved potential N-linked glycan (PNLG) site 479:534 a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 479:603 Previous work has shown that removal of a highly conserved potential N-linked glycan (PNLG) site at amino acid residue 197 (N7) on the surface antigen gp120 of HIV-1 increases neutralization sensitivity of the mutant virus to CD4 binding site (CD4bs)-directed antibodies compared to its wild-type (WT) counterpart. 28255882 0 0 theme Mammalian 77:85 arg1 Cells 87:91 Mammalian Cells 77:91 Mammalian Cells 77:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 3 1 theme constant 499:506 arg1 domains 508:514 α/β TCR constant domains 491:514 α/β TCR constant domains expressed in mammalian cells 491:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 4 2 theme mass 584:587 arg1 mapping 589:595 peptide mass mapping 576:595 peptide mass mapping 576:595 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 3 3 dep based 382:386 arg1 spectrometry 369:380 mass spectrometry 364:380 mass spectrometry 364:380 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 4 4 theme heterogeneity 695:707 arg1 analyses 660:667 analyses 660:667 analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures 660:756 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 4 5 theme peptide 576:582 arg1 mapping 589:595 peptide mass mapping 576:595 peptide mass mapping 576:595 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 3 6 theme mass 364:367 arg1 spectrometry 369:380 mass spectrometry 364:380 mass spectrometry 364:380 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 4 7 theme glycan 723:728 arg1 compositions 730:741 possible glycan compositions 714:741 possible glycan compositions 714:741 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 4 8 theme possible 714:721 arg1 compositions 730:741 possible glycan compositions 714:741 possible glycan compositions 714:741 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 3 9 with proteins 477:484 arg1 domains 508:514 α/β TCR constant domains 491:514 α/β TCR constant domains expressed in mammalian cells 491:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 4 10 theme glycan 632:637 arg1 profiling 639:647 N-linked glycan profiling 623:647 N-linked glycan profiling 623:647 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 1 11 dep proteins 121:128 arg1 secreted 144:151 secreted 144:151 secreted from mammalian cells 144:172 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 1 11 dep proteins 121:128 arg1 expressed 130:138 expressed 130:138 expressed 130:138 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 4 12 theme occupancy 677:685 arg1 analyses 660:667 analyses 660:667 analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures 660:756 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 3 13 theme bottom 405:410 arg1 approach 416:423 a "bottom up" approach 402:423 a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 402:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 3 14 theme " 414:414 arg1 approach 416:423 a "bottom up" approach 402:423 a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 402:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 0 15 theme Glycosylation 0:12 arg1 Profiling 14:22 Glycosylation Profiling 0:22 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 0:92 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 1 16 gly glycoprotein 222:233 arg1 glycoprotein 222:233 glycoprotein receptors 222:243 glycoprotein receptors 222:243 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 4 17 theme site 672:675 arg1 occupancy 677:685 site occupancy 672:685 site occupancy 672:685 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 0 18 theme α/β 27:29 arg1 Domains 56:62 α/β T Cell Receptor Constant Domains 27:62 α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells 27:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 1 19 theme Glycoprofiling 94:107 arg1 proteins 121:128 Glycoprofiling recombinant proteins 94:128 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells 94:172 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 3 20 theme recombinant 458:468 arg1 proteins 477:484 recombinant fusion proteins 458:484 recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 458:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 3 21 theme based 382:386 arg1 protocol 388:395 a mass spectrometry based protocol 362:395 a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 362:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 3 22 theme mammalian 529:537 arg1 cells 539:543 mammalian cells 529:543 mammalian cells 529:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 4 23 link N-linked 623:630 arg1 profiling 639:647 N-linked glycan profiling 623:647 N-linked glycan profiling 623:647 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 1 24 theme recombinant 109:119 arg1 proteins 121:128 Glycoprofiling recombinant proteins 94:128 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells 94:172 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 3 25 theme fusion 470:475 arg1 proteins 477:484 recombinant fusion proteins 458:484 recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 458:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 0 26 theme Cell 33:36 arg1 Domains 56:62 α/β T Cell Receptor Constant Domains 27:62 α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells 27:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 1 27 with interactions 204:215 arg1 receptors 235:243 glycoprotein receptors 222:243 glycoprotein receptors 222:243 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 1 28 theme glycoprotein 222:233 arg1 receptors 235:243 glycoprotein receptors 222:243 glycoprotein receptors 222:243 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 28255882 0 29 theme T 31:31 arg1 Domains 56:62 α/β T Cell Receptor Constant Domains 27:62 α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells 27:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 3 30 theme up 412:413 arg1 approach 416:423 a "bottom up" approach 402:423 a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 402:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 3 31 from glycosylation 441:453 arg1 proteins 477:484 recombinant fusion proteins 458:484 recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 458:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 0 32 theme Constant 47:54 arg1 Domains 56:62 α/β T Cell Receptor Constant Domains 27:62 α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells 27:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 3 33 theme α/β 491:493 arg1 domains 508:514 α/β TCR constant domains 491:514 α/β TCR constant domains expressed in mammalian cells 491:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 0 34 theme Receptor 38:45 arg1 Domains 56:62 α/β T Cell Receptor Constant Domains 27:62 α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells 27:91 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 3 35 gly glycosylation 441:453 arg1 proteins 477:484 recombinant fusion proteins 458:484 recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 458:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 3 36 theme TCR 495:497 arg1 domains 508:514 α/β TCR constant domains 491:514 α/β TCR constant domains expressed in mammalian cells 491:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 2 37 theme cell 278:281 arg1 TCRs 294:297 TCRs 294:297 TCRs 294:297 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 37 theme cell 278:281 arg1 receptors 283:291 recombinant T cell receptors 264:291 recombinant T cell receptors (TCRs) 264:298 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 37 theme cell 278:281 arg1 moieties 336:343 therapeutic moieties 324:343 therapeutic moieties 324:343 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 4 38 theme compositions 730:741 arg1 analyses 660:667 analyses 660:667 analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures 660:756 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 3 39 with protocol 388:395 arg1 approach 416:423 a "bottom up" approach 402:423 a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells 402:543 Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. 28255882 2 40 theme T 276:276 arg1 TCRs 294:297 TCRs 294:297 TCRs 294:297 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 40 theme T 276:276 arg1 receptors 283:291 recombinant T cell receptors 264:291 recombinant T cell receptors (TCRs) 264:298 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 40 theme T 276:276 arg1 moieties 336:343 therapeutic moieties 324:343 therapeutic moieties 324:343 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 41 theme therapeutic 324:334 arg1 receptors 283:291 recombinant T cell receptors 264:291 recombinant T cell receptors (TCRs) 264:298 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 41 theme therapeutic 324:334 arg1 moieties 336:343 therapeutic moieties 324:343 therapeutic moieties 324:343 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 4 42 theme mass 601:604 arg1 spectrometry 606:617 mass spectrometry 601:617 mass spectrometry 601:617 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 2 43 theme recombinant 264:274 arg1 TCRs 294:297 TCRs 294:297 TCRs 294:297 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 43 theme recombinant 264:274 arg1 receptors 283:291 recombinant T cell receptors 264:291 recombinant T cell receptors (TCRs) 264:298 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 2 43 theme recombinant 264:274 arg1 moieties 336:343 therapeutic moieties 324:343 therapeutic moieties 324:343 Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. 28255882 4 44 theme glycan 688:693 arg1 heterogeneity 695:707 glycan heterogeneity 688:707 glycan heterogeneity 688:707 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 4 45 theme N-linked 623:630 arg1 profiling 639:647 N-linked glycan profiling 623:647 N-linked glycan profiling 623:647 The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. 28255882 0 46 theme Domains 56:62 arg1 Profiling 14:22 Glycosylation Profiling 0:22 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 0:92 Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells. 28255882 1 47 theme mammalian 158:166 arg1 cells 168:172 mammalian cells 158:172 mammalian cells 158:172 Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. 27489265 7 0 from interface 1225:1233 arg1 loops 1203:1207 the variable loops 1190:1207 the variable loops at the trimeric interface of adjacent protomers 1190:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 7 0 from interface 1225:1233 arg1 flexibility 1175:1185 the flexibility 1171:1185 the flexibility of the variable loops at the trimeric interface of adjacent protomers 1171:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 6 1 theme N197Q 1003:1007 arg1 mutant 1009:1014 the N197Q mutant 999:1014 the N197Q mutant in trimeric Env 999:1030 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 2 2 theme immune 298:303 arg1 response 305:312 the host's immune response 287:312 the host's immune response 287:312 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 10 3 from immunogens 1764:1773 arg1 vaccines 1778:1785 vaccines 1778:1785 vaccines 1778:1785 These findings present a simple modification for enhancing trimeric Env immunogens in vaccines. 27489265 14 4 theme trimer 2361:2366 arg1 crown 2368:2372 the trimer crown 2357:2372 the trimer crown 2357:2372 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 3 5 from N197 458:461 arg1 removal 414:420 the removal 410:420 the removal of a particular conserved glycan at N197 410:461 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 2 6 theme mechanisms 245:254 arg1 mechanisms 245:254 the mechanisms 241:254 the mechanisms that HIV has evolved to escape the host's immune response 241:312 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 2 6 theme mechanisms 245:254 arg1 One 234:236 One 234:236 One 234:236 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 2 7 with Env 347:349 arg1 glycosylation 362:374 dense glycosylation 356:374 dense glycosylation 356:374 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 7 8 theme mass 1319:1322 arg1 spectrometry 1324:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 3 9 theme vaccine 636:642 arg1 design 644:649 vaccine design 636:649 vaccine design 636:649 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 10 theme neutralization 477:490 arg1 sensitivity 492:502 the neutralization sensitivity 473:502 the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs) 473:568 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 15 11 theme epitope 2530:2536 arg1 presentation 2538:2549 epitope presentation 2530:2549 epitope presentation in Env-based vaccines 2530:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 5 12 theme SOSIP 787:791 arg1 trimers 793:799 native-like SOSIP trimers 775:799 native-like SOSIP trimers 775:799 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 14 13 theme structural 2334:2343 arg1 dynamics 2345:2352 the structural dynamics 2330:2352 the structural dynamics at the trimer crown 2330:2372 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 9 14 theme N197 1568:1571 arg1 glycan 1573:1578 the N197 glycan 1564:1578 the N197 glycan 1564:1578 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 7 15 theme hydrogen-deuterium 1291:1308 arg1 spectrometry 1324:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 8 16 from antibodies 1469:1478 arg1 trimer 1491:1496 the Env trimer 1483:1496 the Env trimer 1483:1496 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 7 17 from flexibility 1175:1185 arg1 interface 1225:1233 the trimeric interface 1212:1233 the trimeric interface of adjacent protomers 1212:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 0 18 theme Binding 101:107 arg1 Glycan 123:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 13 19 theme N-linked 2015:2022 arg1 glycans 2024:2030 many N-linked glycans 2010:2030 many N-linked glycans 2010:2030 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 3 20 theme virus 511:515 arg1 sensitivity 492:502 the neutralization sensitivity 473:502 the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs) 473:568 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 15 21 theme effective 2503:2511 arg1 way 2513:2515 an effective way 2500:2515 an effective way of sculpting epitope presentation in Env-based vaccines 2500:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 21 theme effective 2503:2511 arg1 modification 2480:2491 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 22 gly glycosylation 2461:2473 arg2 site 2475:2478 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 22 gly glycosylation 2461:2473 arg2 modification 2480:2491 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 22 gly glycosylation 2461:2473 arg2 way 2513:2515 an effective way 2500:2515 an effective way of sculpting epitope presentation in Env-based vaccines 2500:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 14 23 dep conserved 2157:2165 arg1 site-proximal 2180:2192 site-proximal 2180:2192 site-proximal 2180:2192 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 0 24 theme Glycan 123:128 arg1 Removal 74:80 Removal 74:80 Removal of a Conserved CD4 Binding Site-Proximal Glycan 74:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 3 25 theme binding 549:555 arg1 CD4bs 563:567 CD4bs 563:567 CD4bs 563:567 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 25 theme binding 549:555 arg1 site 557:560 the CD4 binding site 541:560 the CD4 binding site (CD4bs) 541:568 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 9 26 theme relevant 1605:1612 arg1 epitopes 1619:1626 relevant bNAb epitopes 1605:1626 relevant bNAb epitopes 1605:1626 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 8 27 gly occupancy 1405:1413 arg2 glycan 1427:1432 the N197 glycan 1418:1432 the N197 glycan 1418:1432 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 7 28 from mutant 1281:1286 arg1 increase 1159:1166 a subtle increase 1150:1166 a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers 1150:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 7 28 from mutant 1281:1286 arg1 evident 1260:1266 evident 1260:1266 evident 1260:1266 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 9 29 theme epitopes 1619:1626 arg1 exposure 1593:1600 the exposure 1589:1600 the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure 1589:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 8 30 theme chains 1372:1377 arg1 modeling 1349:1356 Structural modeling 1338:1356 Structural modeling of the glycan chains 1338:1377 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 7 31 theme protomers 1247:1255 arg1 interface 1225:1233 the trimeric interface 1212:1233 the trimeric interface of adjacent protomers 1212:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 1 32 gly glycoprotein 144:155 arg1 target 176:181 the major target 166:181 the major target for HIV-1 166:191 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 1 32 gly glycoprotein 144:155 arg1 Env 158:160 Env 158:160 Env 158:160 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 1 32 gly glycoprotein 144:155 arg1 glycoprotein 144:155 The envelope glycoprotein 131:155 The envelope glycoprotein (Env) 131:161 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 5 33 theme local 845:849 arg1 dynamics 862:869 local structural dynamics 845:869 local structural dynamics 845:869 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 7 34 theme variable 1194:1201 arg1 loops 1203:1207 the variable loops 1190:1207 the variable loops at the trimeric interface of adjacent protomers 1190:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 12 35 theme protective 1938:1947 arg1 mechanism 1949:1957 a major protective mechanism 1930:1957 a major protective mechanism against immune recognition 1930:1984 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 12 35 theme protective 1938:1947 arg1 glycan 1896:1901 This so-called glycan shield 1881:1908 This so-called glycan shield 1881:1908 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 0 36 theme Trimers 51:57 arg1 Structure 11:19 Structure 11:19 Structure 11:19 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 0 36 theme Trimers 51:57 arg1 Antigenicity 25:36 Antigenicity 25:36 Antigenicity 25:36 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 11 37 theme glycans 1872:1878 arg1 patchwork 1832:1840 a dense patchwork 1824:1840 a dense patchwork of host cell-derived N-linked glycans 1824:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 1 38 theme envelope 135:142 arg1 target 176:181 the major target 166:181 the major target for HIV-1 166:191 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 1 38 theme envelope 135:142 arg1 Env 158:160 Env 158:160 Env 158:160 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 1 38 theme envelope 135:142 arg1 glycoprotein 144:155 The envelope glycoprotein 131:155 The envelope glycoprotein (Env) 131:161 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 8 39 theme spatial 1397:1403 arg1 occupancy 1405:1413 the spatial occupancy 1393:1413 the spatial occupancy of the N197 glycan 1393:1432 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 14 40 from crown 2368:2372 arg1 dynamics 2345:2352 the structural dynamics 2330:2352 the structural dynamics at the trimer crown 2330:2372 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 11 41 theme N-linked 1863:1870 arg1 glycans 1872:1878 host cell-derived N-linked glycans 1845:1878 host cell-derived N-linked glycans 1845:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 0 42 from Changes 0:6 arg1 Structure 11:19 Structure 11:19 Structure 11:19 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 0 42 from Changes 0:6 arg1 Antigenicity 25:36 Antigenicity 25:36 Antigenicity 25:36 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 14 43 theme antibody 2269:2276 arg1 access 2278:2283 neutralizing antibody access 2256:2283 neutralizing antibody access to the CD4 binding site 2256:2307 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 8 44 theme N197 1422:1425 arg1 glycan 1427:1432 the N197 glycan 1418:1432 the N197 glycan 1418:1432 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 11 45 theme dense 1826:1830 arg1 patchwork 1832:1840 a dense patchwork 1824:1840 a dense patchwork of host cell-derived N-linked glycans 1824:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 14 46 theme conserved 2157:2165 arg1 glycan 2199:2204 the conserved, CD4 binding site-proximal N197 glycan 2153:2204 the conserved, CD4 binding site-proximal N197 glycan 2153:2204 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 11 47 theme HIV-1 1792:1796 arg1 glycoprotein 1802:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 0 48 theme Conserved 87:95 arg1 Glycan 123:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 9 49 theme trimeric 1672:1679 arg1 structure 1681:1689 the overall trimeric structure 1660:1689 the overall trimeric structure 1660:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 2 50 theme conserved 325:333 arg1 epitopes 335:342 conserved epitopes 325:342 conserved epitopes 325:342 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 13 51 theme functional 2109:2118 arg1 roles 2120:2124 key functional roles 2105:2124 key functional roles 2105:2124 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 7 52 from evident 1260:1266 arg1 mutant 1281:1286 the N197Q mutant 1271:1286 the N197Q mutant by hydrogen-deuterium exchange mass spectrometry 1271:1335 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 6 53 theme antibodies 972:981 arg1 binding 938:944 the binding 934:944 the binding of CD4bs and V3-targeting antibodies 934:981 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 8 54 theme steric 1443:1448 arg1 clashes 1450:1456 steric clashes 1443:1456 steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120 1443:1520 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 4 55 theme structural 668:677 arg1 consequences 679:690 the structural consequences 664:690 the structural consequences that result from the removal of the N197 glycan 664:738 At present, the structural consequences that result from the removal of the N197 glycan have not been characterized. 27489265 6 56 theme monomeric 1068:1076 arg1 gp120 1078:1082 monomeric gp120 1068:1082 monomeric gp120 1068:1082 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 7 57 theme loops 1203:1207 arg1 flexibility 1175:1185 the flexibility 1171:1185 the flexibility of the variable loops at the trimeric interface of adjacent protomers 1171:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 12 58 dep glycan 1896:1901 arg1 shield 1903:1908 shield 1903:1908 shield 1903:1908 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 4 59 theme glycan 733:738 arg1 removal 713:719 the removal 709:719 the removal of the N197 glycan 709:738 At present, the structural consequences that result from the removal of the N197 glycan have not been characterized. 27489265 9 60 from impact 1650:1655 arg1 structure 1681:1689 the overall trimeric structure 1660:1689 the overall trimeric structure 1660:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 3 61 theme particular 427:436 arg1 glycan 448:453 a particular conserved glycan 425:453 a particular conserved glycan 425:453 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 0 62 theme HIV-1 41:45 arg1 Trimers 51:57 HIV-1 Env Trimers 41:57 HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan 41:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 14 63 theme trimer 2414:2419 arg1 stability 2421:2429 global Env trimer stability 2403:2429 global Env trimer stability 2403:2429 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 3 64 theme interest 603:610 arg1 site 583:586 a site 581:586 it a site of significant interest 578:610 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 64 theme interest 603:610 arg1 interest 603:610 significant interest 591:610 significant interest 591:610 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 14 65 theme global 2403:2408 arg1 stability 2421:2429 global Env trimer stability 2403:2429 global Env trimer stability 2403:2429 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 6 66 theme trimeric 1019:1026 arg1 Env 1028:1030 trimeric Env 1019:1030 trimeric Env 1019:1030 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 10 67 theme Env 1760:1762 arg1 immunogens 1764:1773 trimeric Env immunogens 1751:1773 trimeric Env immunogens in vaccines 1751:1785 These findings present a simple modification for enhancing trimeric Env immunogens in vaccines. 27489265 8 68 theme Structural 1338:1347 arg1 modeling 1349:1356 Structural modeling 1338:1356 Structural modeling of the glycan chains 1338:1377 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 14 69 theme binding 2296:2302 arg1 site 2304:2307 the CD4 binding site 2288:2307 the CD4 binding site 2288:2307 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 3 70 dep it 578:579 arg1 site 583:586 a site 581:586 it a site of significant interest 578:610 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 70 dep it 578:579 arg1 interest 603:610 significant interest 591:610 significant interest 591:610 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 7 71 theme subtle 1152:1157 arg1 evident 1260:1266 evident 1260:1266 evident 1260:1266 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 7 71 theme subtle 1152:1157 arg1 increase 1159:1166 a subtle increase 1150:1166 a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers 1150:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 6 72 from mutant 1009:1014 arg1 Env 1028:1030 trimeric Env 1019:1030 trimeric Env 1019:1030 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 3 73 theme glycan 448:453 arg1 removal 414:420 the removal 410:420 the removal of a particular conserved glycan at N197 410:461 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 15 74 theme site 2475:2478 arg1 way 2513:2515 an effective way 2500:2515 an effective way of sculpting epitope presentation in Env-based vaccines 2500:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 74 theme site 2475:2478 arg1 modification 2480:2491 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 3 75 attach removal 414:420 arg2 glycan 448:453 a particular conserved glycan 425:453 a particular conserved glycan 425:453 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 75 attach removal 414:420 arg3 N197 458:461 N197 458:461 N197 458:461 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 15 76 theme surgical 2452:2459 arg1 way 2513:2515 an effective way 2500:2515 an effective way of sculpting epitope presentation in Env-based vaccines 2500:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 76 theme surgical 2452:2459 arg1 modification 2480:2491 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 12 77 theme so-called 1886:1894 arg1 mechanism 1949:1957 a major protective mechanism 1930:1957 a major protective mechanism against immune recognition 1930:1984 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 12 77 theme so-called 1886:1894 arg1 glycan 1896:1901 This so-called glycan shield 1881:1908 This so-called glycan shield 1881:1908 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 14 78 theme neutralizing 2256:2267 arg1 access 2278:2283 neutralizing antibody access 2256:2283 neutralizing antibody access to the CD4 binding site 2256:2307 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 15 79 theme Env-based 2554:2562 arg1 vaccines 2564:2571 Env-based vaccines 2554:2571 Env-based vaccines 2554:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 5 80 theme native-like 775:785 arg1 trimers 793:799 native-like SOSIP trimers 775:799 native-like SOSIP trimers 775:799 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 9 81 from exposure 1593:1600 arg1 Env 1631:1633 Env 1631:1633 Env with a minimal impact on the overall trimeric structure 1631:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 7 82 from increase 1159:1166 arg1 flexibility 1175:1185 the flexibility 1171:1185 the flexibility of the variable loops at the trimeric interface of adjacent protomers 1171:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 3 83 theme design 644:649 arg1 perspective 621:631 the perspective 617:631 the perspective of vaccine design 617:649 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 9 84 theme glycan 1573:1578 arg1 removal 1553:1559 the removal 1549:1559 the removal of the N197 glycan 1549:1578 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 7 85 theme exchange 1310:1317 arg1 spectrometry 1324:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 hydrogen-deuterium exchange mass spectrometry 1291:1335 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 6 86 from increase 922:929 arg1 binding 938:944 the binding 934:944 the binding of CD4bs and V3-targeting antibodies 934:981 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 0 87 theme Site-Proximal 109:121 arg1 Glycan 123:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 1 88 theme major 170:174 arg1 target 176:181 the major target 166:181 the major target for HIV-1 166:191 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 1 88 theme major 170:174 arg1 glycoprotein 144:155 The envelope glycoprotein 131:155 The envelope glycoprotein (Env) 131:161 The envelope glycoprotein (Env) is the major target for HIV-1 broadly neutralizing antibodies (bNAbs). 27489265 13 89 theme glycans 2024:2030 arg1 specific 2044:2051 specific 2044:2051 specific 2044:2051 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 13 89 theme glycans 2024:2030 arg1 positions 1997:2005 the positions 1993:2005 the positions of many N-linked glycans 1993:2030 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 3 90 theme CD4 545:547 arg1 CD4bs 563:567 CD4bs 563:567 CD4bs 563:567 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 90 theme CD4 545:547 arg1 site 557:560 the CD4 binding site 541:560 the CD4 binding site (CD4bs) 541:568 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 8 91 theme glycan 1365:1370 arg1 chains 1372:1377 the glycan chains 1361:1377 the glycan chains 1361:1377 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 7 92 theme N197Q 1275:1279 arg1 mutant 1281:1286 the N197Q mutant 1271:1286 the N197Q mutant by hydrogen-deuterium exchange mass spectrometry 1271:1335 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 8 93 with clashes 1450:1456 arg1 gp120 1516:1520 monomeric gp120 1506:1520 CD4bs antibodies in the Env trimer but not monomeric gp120 1463:1520 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 8 93 with clashes 1450:1456 arg1 antibodies 1469:1478 CD4bs antibodies 1463:1478 CD4bs antibodies in the Env trimer but not monomeric gp120 1463:1520 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 11 94 gly glycoprotein 1802:1813 arg1 glycoprotein 1802:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 9 95 theme bNAb 1614:1617 arg1 epitopes 1619:1626 relevant bNAb epitopes 1605:1626 relevant bNAb epitopes 1605:1626 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 12 96 theme immune 1967:1972 arg1 recognition 1974:1984 immune recognition 1967:1984 immune recognition 1967:1984 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 10 97 theme simple 1717:1722 arg1 modification 1724:1735 a simple modification 1715:1735 a simple modification for enhancing trimeric Env immunogens in vaccines 1715:1785 These findings present a simple modification for enhancing trimeric Env immunogens in vaccines. 27489265 13 98 theme many 2010:2013 arg1 glycans 2024:2030 many N-linked glycans 2010:2030 many N-linked glycans 2010:2030 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 9 99 with Env 1631:1633 arg1 impact 1650:1655 a minimal impact 1640:1655 a minimal impact on the overall trimeric structure 1640:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 7 100 theme adjacent 1238:1245 arg1 protomers 1247:1255 adjacent protomers 1238:1255 adjacent protomers 1238:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 12 101 theme major 1932:1936 arg1 mechanism 1949:1957 a major protective mechanism 1930:1957 a major protective mechanism against immune recognition 1930:1984 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 12 101 theme major 1932:1936 arg1 glycan 1896:1901 This so-called glycan shield 1881:1908 This so-called glycan shield 1881:1908 This so-called glycan shield is considered to be a major protective mechanism against immune recognition. 27489265 14 102 theme N197 2194:2197 arg1 glycan 2199:2204 the conserved, CD4 binding site-proximal N197 glycan 2153:2204 the conserved, CD4 binding site-proximal N197 glycan 2153:2204 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 6 103 theme large 916:920 arg1 increase 922:929 A large increase 914:929 A large increase in the binding of CD4bs and V3-targeting antibodies 914:981 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 9 104 theme minimal 1642:1648 arg1 impact 1650:1655 a minimal impact 1640:1655 a minimal impact on the overall trimeric structure 1640:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 2 105 gly glycosylation 362:374 arg1 Env 347:349 Env 347:349 Env with dense glycosylation 347:374 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 13 106 link N-linked 2015:2022 arg1 glycans 2024:2030 many N-linked glycans 2010:2030 many N-linked glycans 2010:2030 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 7 107 theme overall 1095:1101 arg1 structure 1103:1111 the overall structure 1091:1111 the overall structure 1091:1111 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 5 108 from effects 817:823 arg1 antigenicity 828:839 antigenicity 828:839 antigenicity 828:839 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 5 108 from effects 817:823 arg1 dynamics 862:869 local structural dynamics 845:869 local structural dynamics 845:869 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 8 109 theme glycan 1427:1432 arg1 occupancy 1405:1413 the spatial occupancy 1393:1413 the spatial occupancy of the N197 glycan 1393:1432 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 0 110 theme CD4 97:99 arg1 Glycan 123:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 a Conserved CD4 Binding Site-Proximal Glycan 85:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 7 111 theme trimeric 1216:1223 arg1 interface 1225:1233 the trimeric interface 1212:1233 the trimeric interface of adjacent protomers 1212:1255 While the overall structure and thermostability are not altered, a subtle increase in the flexibility of the variable loops at the trimeric interface of adjacent protomers is evident in the N197Q mutant by hydrogen-deuterium exchange mass spectrometry. 27489265 9 112 theme overall 1664:1670 arg1 structure 1681:1689 the overall trimeric structure 1660:1689 the overall trimeric structure 1660:1689 Our results indicate that the removal of the N197 glycan enhances the exposure of relevant bNAb epitopes on Env with a minimal impact on the overall trimeric structure. 27489265 11 113 theme Env 1798:1800 arg1 glycoprotein 1802:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein 1788:1813 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 5 114 theme structural 851:860 arg1 dynamics 862:869 local structural dynamics 845:869 local structural dynamics 845:869 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 3 115 theme Previous 377:384 arg1 studies 386:392 Previous studies 377:392 Previous studies 377:392 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 11 116 theme host 1845:1848 arg1 glycans 1872:1878 host cell-derived N-linked glycans 1845:1878 host cell-derived N-linked glycans 1845:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 6 117 theme V3-targeting 959:970 arg1 antibodies 972:981 V3-targeting antibodies 959:981 V3-targeting antibodies 959:981 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 8 118 theme CD4bs 1463:1467 arg1 antibodies 1469:1478 CD4bs antibodies 1463:1478 CD4bs antibodies in the Env trimer but not monomeric gp120 1463:1520 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 10 119 theme trimeric 1751:1758 arg1 immunogens 1764:1773 trimeric Env immunogens 1751:1773 trimeric Env immunogens in vaccines 1751:1785 These findings present a simple modification for enhancing trimeric Env immunogens in vaccines. 27489265 2 120 theme dense 356:360 arg1 glycosylation 362:374 dense glycosylation 356:374 dense glycosylation 356:374 One of the mechanisms that HIV has evolved to escape the host's immune response is to mask conserved epitopes on Env with dense glycosylation. 27489265 6 121 theme CD4bs 949:953 arg1 binding 938:944 the binding 934:944 the binding of CD4bs and V3-targeting antibodies 934:981 A large increase in the binding of CD4bs and V3-targeting antibodies is observed for the N197Q mutant in trimeric Env, while no changes are observed with monomeric gp120. 27489265 4 122 theme N197 728:731 arg1 glycan 733:738 the N197 glycan 724:738 the N197 glycan 724:738 At present, the structural consequences that result from the removal of the N197 glycan have not been characterized. 27489265 0 123 theme Env 47:49 arg1 Trimers 51:57 HIV-1 Env Trimers 41:57 HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan 41:128 Changes in Structure and Antigenicity of HIV-1 Env Trimers Resulting from Removal of a Conserved CD4 Binding Site-Proximal Glycan. 27489265 11 124 link cell-derived 1850:1861 arg1 glycans 1872:1878 host cell-derived N-linked glycans 1845:1878 host cell-derived N-linked glycans 1845:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 8 125 theme Env 1487:1489 arg1 trimer 1491:1496 the Env trimer 1483:1496 the Env trimer 1483:1496 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 3 126 theme significant 591:601 arg1 interest 603:610 significant interest 591:610 significant interest 591:610 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 3 127 theme conserved 438:446 arg1 glycan 448:453 a particular conserved glycan 425:453 a particular conserved glycan 425:453 Previous studies have shown that the removal of a particular conserved glycan at N197 increases the neutralization sensitivity of the virus to antibodies targeting the CD4 binding site (CD4bs), making it a site of significant interest from the perspective of vaccine design. 27489265 13 128 theme key 2105:2107 arg1 roles 2120:2124 key functional roles 2105:2124 key functional roles 2105:2124 While the positions of many N-linked glycans are isolate specific, some are highly conserved and are believed to play key functional roles. 27489265 14 129 theme Env 2410:2412 arg1 stability 2421:2429 global Env trimer stability 2403:2429 global Env trimer stability 2403:2429 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 5 130 theme glycan 906:911 arg1 removal 890:896 the removal 886:896 the removal of this glycan 886:911 Using native-like SOSIP trimers, we examine the effects on antigenicity and local structural dynamics resulting from the removal of this glycan. 27489265 11 131 theme cell-derived 1850:1861 arg1 glycans 1872:1878 host cell-derived N-linked glycans 1845:1878 host cell-derived N-linked glycans 1845:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 15 132 from presentation 2538:2549 arg1 vaccines 2564:2571 Env-based vaccines 2554:2571 Env-based vaccines 2554:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 8 133 theme monomeric 1506:1514 arg1 gp120 1516:1520 monomeric gp120 1506:1520 CD4bs antibodies in the Env trimer but not monomeric gp120 1463:1520 Structural modeling of the glycan chains suggests that the spatial occupancy of the N197 glycan leads to steric clashes with CD4bs antibodies in the Env trimer but not monomeric gp120. 27489265 14 134 theme CD4 2292:2294 arg1 site 2304:2307 the CD4 binding site 2288:2307 the CD4 binding site 2288:2307 In this study, we examine the conserved, CD4 binding site-proximal N197 glycan and demonstrate that its removal both facilitates neutralizing antibody access to the CD4 binding site and modestly impacts the structural dynamics at the trimer crown without drastically altering global Env trimer stability. 27489265 11 135 link N-linked 1863:1870 arg1 glycans 1872:1878 host cell-derived N-linked glycans 1845:1878 host cell-derived N-linked glycans 1845:1878 The HIV-1 Env glycoprotein presents a dense patchwork of host cell-derived N-linked glycans. 27489265 15 136 theme glycosylation 2461:2473 arg1 way 2513:2515 an effective way 2500:2515 an effective way of sculpting epitope presentation in Env-based vaccines 2500:2571 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 27489265 15 136 theme glycosylation 2461:2473 arg1 modification 2480:2491 surgical glycosylation site modification 2452:2491 surgical glycosylation site modification 2452:2491 This indicates that surgical glycosylation site modification may be an effective way of sculpting epitope presentation in Env-based vaccines. 28452462 7 0 theme site-selective 1708:1721 arg1 modification 1737:1748 the site-selective glycosylation modification 1704:1748 the site-selective glycosylation modification of the protein 1704:1763 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 8 1 theme multiple 1892:1899 arg1 N-glycans 1901:1909 multiple N-glycans 1892:1909 multiple N-glycans 1892:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 1 2 gly glycoproteins 193:205 arg1 glycoproteins 193:205 glycoproteins 193:205 glycoproteins 193:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 4 3 theme core-fucosylated 1032:1047 arg1 acceptor 1073:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor 1028:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation 1028:1113 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 6 4 theme detailed 1487:1494 arg1 analysis 1502:1509 a detailed MS/MS analysis 1485:1509 a detailed MS/MS analysis of the transglycosylation product 1485:1543 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 3 5 contain contains 824:831 arg2 N-glycans 839:847 three N-glycans 833:847 three N-glycans 833:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 3 5 contain contains 824:831 arg1 remodeling 766:775 site-selective chemoenzymatic glycosylation remodeling 722:775 site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans 722:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 7 6 theme distinct 1667:1674 arg1 oxazoline 1683:1691 a distinct glycan oxazoline 1665:1691 a distinct glycan oxazoline 1665:1691 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 1 7 gly heterogeneity 157:169 arg1 N-glycosylation 174:188 N-glycosylation 174:188 N-glycosylation of glycoproteins 174:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 1 8 theme great 215:219 arg1 challenge 221:229 a great challenge 213:229 a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics 213:339 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 6 9 theme Asn-83 1448:1453 arg1 sites 1455:1459 the Asn-38 and Asn-83 sites 1433:1459 the Asn-38 and Asn-83 sites 1433:1459 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 3 10 theme glycosylation 752:764 arg1 remodeling 766:775 site-selective chemoenzymatic glycosylation remodeling 722:775 site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans 722:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 4 11 theme intermediate 1060:1071 arg1 acceptor 1073:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor 1028:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation 1028:1113 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 6 12 theme product 1537:1543 arg1 analysis 1502:1509 a detailed MS/MS analysis 1485:1509 a detailed MS/MS analysis of the transglycosylation product 1485:1543 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 3 13 theme site-selective 722:735 arg1 remodeling 766:775 site-selective chemoenzymatic glycosylation remodeling 722:775 site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans 722:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 1 14 gly N-glycosylation 174:188 arg1 glycoproteins 193:205 glycoproteins 193:205 glycoproteins 193:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 3 15 theme human 792:796 arg1 EPO 814:816 EPO 814:816 EPO 814:816 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 3 15 theme human 792:796 arg1 erythropoietin 798:811 recombinant human erythropoietin 780:811 recombinant human erythropoietin (EPO) 780:817 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 1 16 theme structural 146:155 arg1 heterogeneity 157:169 The tremendous structural heterogeneity 131:169 The tremendous structural heterogeneity of N-glycosylation of glycoproteins 131:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 0 17 theme Mammalian 65:73 arg1 Engineering 80:90 Combined Mammalian Cell Engineering 56:90 Combined Mammalian Cell Engineering 56:90 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 2 18 theme multiple 664:671 arg1 N-glycans 673:681 multiple N-glycans 664:681 multiple N-glycans 664:681 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 4 19 theme overexpressing 912:925 arg1 line 932:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 4 20 theme line 932:935 arg1 generation 868:877 the generation 864:877 the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line 864:935 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 2 21 theme intact 478:483 arg1 antibodies 485:494 intact antibodies 478:494 intact antibodies 478:494 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 0 22 theme Chemoenzymatic 96:109 arg1 Transglycosylation 111:128 Chemoenzymatic Transglycosylation 96:128 Chemoenzymatic Transglycosylation 96:128 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 5 23 theme azide-tagged 1172:1183 arg1 glycoform 1185:1193 azide-tagged glycoform 1172:1193 azide-tagged glycoform 1172:1193 With this acceptor, homogeneous sialylated glycoform or azide-tagged glycoform were produced using the glycosynthase (EndoF3-D165A) catalyzed transglycosylation. 28452462 7 24 theme different 1557:1565 arg1 N-glycan 1567:1574 a different N-glycan 1555:1574 a different N-glycan 1555:1574 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 2 25 theme N-glycoproteins 452:466 arg1 glycoforms 438:447 homogeneous glycoforms 426:447 homogeneous glycoforms of N-glycoproteins including intact antibodies 426:494 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 0 26 theme Glycan 0:5 arg1 Remodeling 7:16 Glycan Remodeling 0:16 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 0:129 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 1 27 theme protein-based 314:326 arg1 therapeutics 328:339 protein-based therapeutics 314:339 protein-based therapeutics 314:339 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 2 28 theme endoglycosidase-catalyzed 514:538 arg1 deglycosylation 540:554 endoglycosidase-catalyzed deglycosylation 514:554 endoglycosidase-catalyzed deglycosylation 514:554 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 4 29 theme knockout 898:905 arg1 line 932:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 4 30 gly glycoform 989:997 arg1 Man5GlcNAc2Fuc 974:987 an unusual Man5GlcNAc2Fuc 963:987 an unusual Man5GlcNAc2Fuc glycoform 963:997 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 5 31 theme homogeneous 1136:1146 arg1 glycoform 1159:1167 homogeneous sialylated glycoform 1136:1167 homogeneous sialylated glycoform 1136:1167 With this acceptor, homogeneous sialylated glycoform or azide-tagged glycoform were produced using the glycosynthase (EndoF3-D165A) catalyzed transglycosylation. 28452462 8 32 theme first 1792:1796 arg1 example 1798:1804 the first example 1788:1804 the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1788:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 4 33 theme unusual 966:972 arg1 Man5GlcNAc2Fuc 974:987 an unusual Man5GlcNAc2Fuc 963:987 an unusual Man5GlcNAc2Fuc glycoform 963:997 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 0 34 theme Erythropoietin 27:40 arg1 Remodeling 7:16 Glycan Remodeling 0:16 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 0:129 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 7 35 theme protein 1757:1763 arg1 modification 1737:1748 the site-selective glycosylation modification 1704:1748 the site-selective glycosylation modification of the protein 1704:1763 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 2 36 theme glycosynthase-catalyzed 566:588 arg1 transglycosylation 590:607 novel glycosynthase-catalyzed transglycosylation 560:607 novel glycosynthase-catalyzed transglycosylation 560:607 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 4 37 theme glycoform 989:997 arg1 production 949:958 the production 945:958 the production 945:958 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 8 38 gly glycoproteins 1869:1881 arg1 glycoproteins 1869:1881 complex glycoproteins 1861:1881 complex glycoproteins carrying multiple N-glycans 1861:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 8 39 theme chemoenzymatic 1824:1837 arg1 engineering 1846:1856 site-selective chemoenzymatic glycan engineering 1809:1856 site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1809:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 2 40 theme glycan 387:392 arg1 method 405:410 a chemoenzymatic glycan remodeling method 370:410 a chemoenzymatic glycan remodeling method 370:410 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 1 41 theme biological 251:260 arg1 functions 262:270 the biological functions 247:270 the biological functions of specific glycoforms 247:293 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 7 42 theme glycosylation 1723:1735 arg1 modification 1737:1748 the site-selective glycosylation modification 1704:1748 the site-selective glycosylation modification of the protein 1704:1763 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 8 43 theme engineering 1846:1856 arg1 example 1798:1804 the first example 1788:1804 the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1788:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 1 44 dep challenge 221:229 arg1 deciphering 235:245 deciphering 235:245 deciphering the biological functions of specific glycoforms 235:293 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 1 44 dep challenge 221:229 arg1 developing 303:312 developing 303:312 developing protein-based therapeutics 303:339 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 8 45 theme complex 1861:1867 arg1 glycoproteins 1869:1881 complex glycoproteins 1861:1881 complex glycoproteins carrying multiple N-glycans 1861:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 1 46 theme glycoforms 284:293 arg1 functions 262:270 the biological functions 247:270 the biological functions of specific glycoforms 247:293 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 6 47 theme MS/MS 1496:1500 arg1 analysis 1502:1509 a detailed MS/MS analysis 1485:1509 a detailed MS/MS analysis of the transglycosylation product 1485:1543 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 7 48 theme glycan 1676:1681 arg1 oxazoline 1683:1691 a distinct glycan oxazoline 1665:1691 a distinct glycan oxazoline 1665:1691 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 6 49 theme N-glycans 1408:1416 arg1 introduction 1388:1399 the introduction 1384:1399 the introduction of two N-glycans 1384:1416 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 1 50 theme glycoproteins 193:205 arg1 N-glycosylation 174:188 N-glycosylation 174:188 N-glycosylation of glycoproteins 174:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 7 51 theme enzymatic 1631:1639 arg1 transglycosylation 1641:1658 the enzymatic transglycosylation 1627:1658 the enzymatic transglycosylation with a distinct glycan oxazoline 1627:1691 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 4 52 theme enzymatic 1086:1094 arg1 transglycosylation 1096:1113 enzymatic transglycosylation 1086:1113 enzymatic transglycosylation 1086:1113 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 6 53 located observed 1327:1334 arg1 sites 1455:1459 the Asn-38 and Asn-83 sites 1433:1459 the Asn-38 and Asn-83 sites 1433:1459 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 6 53 located observed 1327:1334 arg1 reactions 1362:1370 the transglycosylation reactions 1339:1370 the transglycosylation reactions 1339:1370 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 6 53 located observed 1327:1334 arg2 site-selectivity 1306:1321 a remarkable site-selectivity 1293:1321 a remarkable site-selectivity 1293:1321 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 3 54 theme chemoenzymatic 737:750 arg1 remodeling 766:775 site-selective chemoenzymatic glycosylation remodeling 722:775 site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans 722:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 4 55 gly core-fucosylated 1032:1047 arg1 acceptor 1073:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor 1028:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation 1028:1113 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 4 56 theme GlcNAc-EPO 1049:1058 arg1 acceptor 1073:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor 1028:1080 the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation 1028:1113 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 7 57 theme third 1596:1600 arg1 site 1611:1614 the third (Asn-24) site 1592:1614 the third (Asn-24) site 1592:1614 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 7 57 theme third 1596:1600 arg1 Asn-24 1603:1608 Asn-24 1603:1608 Asn-24 1603:1608 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 6 58 theme transglycosylation 1518:1535 arg1 product 1537:1543 the transglycosylation product 1514:1543 the transglycosylation product 1514:1543 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 3 59 theme recombinant 780:790 arg1 EPO 814:816 EPO 814:816 EPO 814:816 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 3 59 theme recombinant 780:790 arg1 erythropoietin 798:811 recombinant human erythropoietin 780:811 recombinant human erythropoietin (EPO) 780:817 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 1 60 theme tremendous 135:144 arg1 heterogeneity 157:169 The tremendous structural heterogeneity 131:169 The tremendous structural heterogeneity of N-glycosylation of glycoproteins 131:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 6 61 theme Asn-38 1437:1442 arg1 sites 1455:1459 the Asn-38 and Asn-83 sites 1433:1459 the Asn-38 and Asn-83 sites 1433:1459 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 7 62 attach attached 1580:1587 arg2 N-glycan 1567:1574 a different N-glycan 1555:1574 a different N-glycan 1555:1574 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 7 62 attach attached 1580:1587 arg3 site 1611:1614 the third (Asn-24) site 1592:1614 the third (Asn-24) site 1592:1614 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 7 62 attach attached 1580:1587 arg3 Asn-24 1603:1608 Asn-24 1603:1608 Asn-24 1603:1608 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 0 63 theme Combined 56:63 arg1 Engineering 80:90 Combined Mammalian Cell Engineering 56:90 Combined Mammalian Cell Engineering 56:90 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 5 64 gly sialylated 1148:1157 arg1 glycoform 1159:1167 homogeneous sialylated glycoform 1136:1167 homogeneous sialylated glycoform 1136:1167 With this acceptor, homogeneous sialylated glycoform or azide-tagged glycoform were produced using the glycosynthase (EndoF3-D165A) catalyzed transglycosylation. 28452462 7 65 mod modification 1737:1748 arg3 glycosylation 1723:1735 the site-selective glycosylation modification 1704:1748 the site-selective glycosylation modification of the protein 1704:1763 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 7 65 mod modification 1737:1748 arg1 protein 1757:1763 the protein 1753:1763 the protein 1753:1763 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 0 66 theme Cell 75:78 arg1 Engineering 80:90 Combined Mammalian Cell Engineering 56:90 Combined Mammalian Cell Engineering 56:90 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 1 67 theme N-glycosylation 174:188 arg1 heterogeneity 157:169 The tremendous structural heterogeneity 131:169 The tremendous structural heterogeneity of N-glycosylation of glycoproteins 131:205 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28452462 5 68 theme sialylated 1148:1157 arg1 glycoform 1159:1167 homogeneous sialylated glycoform 1136:1167 homogeneous sialylated glycoform 1136:1167 With this acceptor, homogeneous sialylated glycoform or azide-tagged glycoform were produced using the glycosynthase (EndoF3-D165A) catalyzed transglycosylation. 28452462 8 69 contain carrying 1883:1890 arg2 N-glycans 1901:1909 multiple N-glycans 1892:1909 multiple N-glycans 1892:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 8 69 contain carrying 1883:1890 arg1 glycoproteins 1869:1881 complex glycoproteins 1861:1881 complex glycoproteins carrying multiple N-glycans 1861:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 4 70 theme FUT8 907:910 arg1 line 932:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 6 71 theme remarkable 1295:1304 arg1 site-selectivity 1306:1321 a remarkable site-selectivity 1293:1321 a remarkable site-selectivity 1293:1321 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 2 72 theme complex 633:639 arg1 glycoproteins 641:653 complex glycoproteins 633:653 complex glycoproteins carrying multiple N-glycans 633:681 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 4 73 theme cell 927:930 arg1 line 932:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 a HEK293S GnT I knockout FUT8 overexpressing cell line 882:935 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 2 74 gly N-glycoproteins 452:466 arg1 N-glycoproteins 452:466 N-glycoproteins 452:466 N-glycoproteins including intact antibodies 452:494 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 0 75 theme Human 21:25 arg1 EPO 43:45 EPO 43:45 EPO 43:45 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 0 75 theme Human 21:25 arg1 Erythropoietin 27:40 Human Erythropoietin 21:40 Human Erythropoietin (EPO) 21:46 Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation. 28452462 2 76 gly glycoforms 438:447 arg1 N-glycoproteins 452:466 N-glycoproteins 452:466 N-glycoproteins including intact antibodies 452:494 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 8 77 theme site-selective 1809:1822 arg1 engineering 1846:1856 site-selective chemoenzymatic glycan engineering 1809:1856 site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1809:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 3 78 theme erythropoietin 798:811 arg1 remodeling 766:775 site-selective chemoenzymatic glycosylation remodeling 722:775 site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans 722:847 We report here site-selective chemoenzymatic glycosylation remodeling of recombinant human erythropoietin (EPO) that contains three N-glycans. 28452462 7 79 with transglycosylation 1641:1658 arg1 oxazoline 1683:1691 a distinct glycan oxazoline 1665:1691 a distinct glycan oxazoline 1665:1691 Finally, a different N-glycan was attached at the third (Asn-24) site by pushing the enzymatic transglycosylation with a distinct glycan oxazoline, achieving the site-selective glycosylation modification of the protein. 28452462 2 80 theme novel 560:564 arg1 transglycosylation 590:607 novel glycosynthase-catalyzed transglycosylation 560:607 novel glycosynthase-catalyzed transglycosylation 560:607 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 8 81 theme glycan 1839:1844 arg1 engineering 1846:1856 site-selective chemoenzymatic glycan engineering 1809:1856 site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1809:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 4 82 theme Man5GlcNAc2Fuc 974:987 arg1 glycoform 989:997 an unusual Man5GlcNAc2Fuc glycoform 963:997 an unusual Man5GlcNAc2Fuc glycoform 963:997 We found that the generation of a HEK293S GnT I knockout FUT8 overexpressing cell line enabled the production of an unusual Man5GlcNAc2Fuc glycoform, which could be converted to the core-fucosylated GlcNAc-EPO intermediate acceptor for enzymatic transglycosylation. 28452462 2 83 theme remodeling 394:403 arg1 method 405:410 a chemoenzymatic glycan remodeling method 370:410 a chemoenzymatic glycan remodeling method 370:410 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 6 84 theme transglycosylation 1343:1360 arg1 reactions 1362:1370 the transglycosylation reactions 1339:1370 the transglycosylation reactions 1339:1370 Interestingly, a remarkable site-selectivity was observed in the transglycosylation reactions, leading to the introduction of two N-glycans selectively at the Asn-38 and Asn-83 sites, which was confirmed by a detailed MS/MS analysis of the transglycosylation product. 28452462 2 85 gly glycoproteins 641:653 arg1 glycoproteins 641:653 complex glycoproteins 633:653 complex glycoproteins carrying multiple N-glycans 633:681 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 2 86 theme chemoenzymatic 372:385 arg1 method 405:410 a chemoenzymatic glycan remodeling method 370:410 a chemoenzymatic glycan remodeling method 370:410 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 8 87 theme glycoproteins 1869:1881 arg1 engineering 1846:1856 site-selective chemoenzymatic glycan engineering 1809:1856 site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans 1809:1909 This study represents the first example of site-selective chemoenzymatic glycan engineering of complex glycoproteins carrying multiple N-glycans. 28452462 2 88 contain carrying 655:662 arg1 glycoproteins 641:653 complex glycoproteins 633:653 complex glycoproteins carrying multiple N-glycans 633:681 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 2 88 contain carrying 655:662 arg2 N-glycans 673:681 multiple N-glycans 664:681 multiple N-glycans 664:681 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 2 89 theme homogeneous 426:436 arg1 glycoforms 438:447 homogeneous glycoforms 426:447 homogeneous glycoforms of N-glycoproteins including intact antibodies 426:494 We have previously reported a chemoenzymatic glycan remodeling method for producing homogeneous glycoforms of N-glycoproteins including intact antibodies, which consist of endoglycosidase-catalyzed deglycosylation and novel glycosynthase-catalyzed transglycosylation, but its application to complex glycoproteins carrying multiple N-glycans remains to be examined. 28452462 1 90 theme specific 275:282 arg1 glycoforms 284:293 specific glycoforms 275:293 specific glycoforms 275:293 The tremendous structural heterogeneity of N-glycosylation of glycoproteins poses a great challenge for deciphering the biological functions of specific glycoforms and for developing protein-based therapeutics. 28714086 9 0 theme protein-protein 1683:1697 arg1 interactions 1699:1710 many protein-protein interactions 1678:1710 many protein-protein interactions 1678:1710 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 3 1 attach attached 490:497 arg3 sites 502:506 sites 502:506 sites along iGluRs that position them for involvement in the structural changes underlying gating 502:598 Glycan moieties are large, polar and occasionally charged, and they are attached at sites along iGluRs that position them for involvement in the structural changes underlying gating. 28714086 3 1 attach attached 490:497 arg2 they 481:484 they 481:484 they 481:484 Glycan moieties are large, polar and occasionally charged, and they are attached at sites along iGluRs that position them for involvement in the structural changes underlying gating. 28714086 7 2 theme consensus 1259:1267 arg1 sites 1269:1273 four and 12 consensus sites 1247:1273 sites 1269:1273 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 2 3 theme binding 370:376 arg1 site 378:381 a binding site 368:381 a binding site for carbohydrate binding proteins 368:415 N-glycosylation is crucial for receptor biogenesis, influences receptor trafficking and provides a binding site for carbohydrate binding proteins. 28714086 7 4 from sites 1269:1273 arg1 subunit 1303:1309 each subunit 1298:1309 each subunit 1298:1309 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 9 5 theme Glycan 1611:1616 arg1 moieties 1618:1625 Glycan moieties 1611:1625 Glycan moieties 1611:1625 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 9 6 theme nervous 1727:1733 arg1 system 1735:1740 the nervous system 1723:1740 the nervous system 1723:1740 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 1 7 theme Golgi 164:168 arg1 processing 170:179 Golgi processing 164:179 Golgi processing 164:179 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 11 8 theme modified 2073:2080 arg1 sugars 2082:2087 both the modified sugars 2064:2087 both the modified sugars 2064:2087 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 5 9 theme KAR 949:951 arg1 properties 964:973 recombinant KAR functional properties 937:973 recombinant KAR functional properties 937:973 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 8 10 from folding 1469:1475 arg1 cells 1500:1504 mammalian cells 1490:1504 mammalian cells 1490:1504 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 5 11 theme trisaccharide 846:858 arg1 HNK-1 860:864 the charged trisaccharide HNK-1 834:864 the charged trisaccharide HNK-1 834:864 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 11 12 theme sugars 2082:2087 arg1 identity 2052:2059 the identity 2048:2059 the identity of both the modified sugars 2048:2087 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 12 theme sugars 2082:2087 arg1 composition 2105:2115 the subunit composition 2093:2115 the subunit composition of the receptor 2093:2131 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 13 theme kainate 1947:1953 arg1 receptors 1955:1963 kainate receptors 1947:1963 kainate receptors (KARs) 1947:1970 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 13 theme kainate 1947:1953 arg1 KARs 1966:1969 KARs 1966:1969 KARs 1966:1969 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 4 14 theme receptor 714:721 arg1 properties 696:705 functional properties 685:705 functional properties 685:705 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 14 theme receptor 714:721 arg1 recovery 749:756 recovery 749:756 recovery from desensitization 749:777 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 14 theme receptor 714:721 arg1 desensitization 732:746 desensitization 732:746 desensitization 732:746 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 14 theme receptor 714:721 arg1 deactivation 783:794 deactivation 783:794 deactivation 783:794 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 10 15 theme structural 1847:1856 arg1 changes 1858:1864 the structural changes 1843:1864 the structural changes underlying gating 1843:1882 Additionally, they are attached at sites along iGluR subunits that position them for involvement in the structural changes underlying gating. 28714086 12 16 dep oligosaccharide 2219:2233 arg1 killer-1 2249:2256 human natural killer-1 2235:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 7 17 theme structural 1358:1367 arg1 diversity 1369:1377 structural diversity 1358:1377 structural diversity conferred by this post-translational modification 1358:1427 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 4 18 theme functional 685:694 arg1 properties 696:705 functional properties 685:705 functional properties 685:705 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 18 theme functional 685:694 arg1 recovery 749:756 recovery 749:756 recovery from desensitization 749:777 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 18 theme functional 685:694 arg1 desensitization 732:746 desensitization 732:746 desensitization 732:746 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 18 theme functional 685:694 arg1 deactivation 783:794 deactivation 783:794 deactivation 783:794 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 7 19 theme 12 1256:1257 arg1 sites 1269:1273 four and 12 consensus sites 1247:1273 sites 1269:1273 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 1 20 theme 4-12 148:151 arg1 sites 153:157 4-12 sites 148:157 4-12 sites 148:157 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 10 21 from involvement 1828:1838 arg1 changes 1858:1864 the structural changes 1843:1864 the structural changes underlying gating 1843:1882 Additionally, they are attached at sites along iGluR subunits that position them for involvement in the structural changes underlying gating. 28714086 8 22 theme oligosaccharides 1530:1545 arg1 clear 1604:1608 clear 1604:1608 clear 1604:1608 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 8 22 theme oligosaccharides 1530:1545 arg1 impact 1520:1525 the impact 1516:1525 the impact of oligosaccharides on the function of successfully folded receptors 1516:1594 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 3 23 from involvement 544:554 arg1 changes 574:580 the structural changes 559:580 the structural changes underlying gating 559:598 Glycan moieties are large, polar and occasionally charged, and they are attached at sites along iGluRs that position them for involvement in the structural changes underlying gating. 28714086 7 24 theme four 1247:1250 arg1 sites 1269:1273 four and 12 consensus sites 1247:1273 sites 1269:1273 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 1 25 from sites 153:157 arg1 N-glycosylated 130:143 N-glycosylated 130:143 N-glycosylated 130:143 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 6 26 theme molecular 1002:1010 arg1 composition 1012:1022 the molecular composition 998:1022 the molecular composition of N-glycans 998:1035 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 8 27 from impact 1520:1525 arg1 function 1554:1561 the function 1550:1561 the function of successfully folded receptors 1550:1594 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 4 28 from desensitization 763:777 arg1 recovery 749:756 recovery 749:756 recovery from desensitization 749:777 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 28 from desensitization 763:777 arg1 desensitization 732:746 desensitization 732:746 desensitization 732:746 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 28 from desensitization 763:777 arg1 deactivation 783:794 deactivation 783:794 deactivation 783:794 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 7 29 theme tetrameric 1214:1223 arg1 proteins 1225:1232 tetrameric proteins 1214:1232 tetrameric proteins 1214:1232 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 29 theme tetrameric 1214:1223 arg1 ABSTRACT 1160:1167 ABSTRACT 1160:1167 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 1160:1428 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 29 theme tetrameric 1214:1223 arg1 receptors 1191:1199 Ionotropic glutamate receptors 1170:1199 Ionotropic glutamate receptors (iGluRs) 1170:1208 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 0 30 theme functional 44:53 arg1 properties 55:64 kainate receptor functional properties 27:64 kainate receptor functional properties 27:64 N-glycan content modulates kainate receptor functional properties. 28714086 1 31 theme Ionotropic 79:88 arg1 receptor 100:107 Ionotropic glutamate receptor 79:107 Ionotropic glutamate receptor (iGluR) subunits 79:124 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 31 theme Ionotropic 79:88 arg1 iGluR 110:114 iGluR 110:114 iGluR 110:114 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 6 32 theme potential 1091:1099 arg1 mechanism 1101:1109 a potential mechanism 1089:1109 a potential mechanism for fine-tuning the function of these receptors 1089:1157 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 8 33 theme folded 1579:1584 arg1 receptors 1586:1594 successfully folded receptors 1566:1594 successfully folded receptors 1566:1594 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 12 34 theme human 2235:2239 arg1 killer-1 2249:2256 human natural killer-1 2235:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 0 35 theme N-glycan 0:7 arg1 content 9:15 N-glycan content 0:15 N-glycan content 0:15 N-glycan content modulates kainate receptor functional properties. 28714086 1 36 theme receptor 100:107 arg1 subunits 117:124 Ionotropic glutamate receptor (iGluR) subunits 79:124 Ionotropic glutamate receptor (iGluR) subunits 79:124 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 7 37 theme high 1343:1346 arg1 degree 1348:1353 a high degree 1341:1353 a high degree of structural diversity conferred by this post-translational modification 1341:1427 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 13 38 from fine-tuning 2430:2440 arg1 structure 2486:2494 glycan structure 2479:2494 glycan structure 2479:2494 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 0 39 theme receptor 35:42 arg1 properties 55:64 kainate receptor functional properties 27:64 kainate receptor functional properties 27:64 N-glycan content modulates kainate receptor functional properties. 28714086 7 40 theme glutamate 1181:1189 arg1 proteins 1225:1232 tetrameric proteins 1214:1232 tetrameric proteins 1214:1232 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 40 theme glutamate 1181:1189 arg1 ABSTRACT 1160:1167 ABSTRACT 1160:1167 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 1160:1428 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 40 theme glutamate 1181:1189 arg1 iGluRs 1202:1207 iGluRs 1202:1207 iGluRs 1202:1207 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 40 theme glutamate 1181:1189 arg1 receptors 1191:1199 Ionotropic glutamate receptors 1170:1199 Ionotropic glutamate receptors (iGluRs) 1170:1208 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 6 41 theme KAR 1051:1053 arg1 properties 1067:1076 KAR biophysical properties 1051:1076 KAR biophysical properties 1051:1076 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 12 42 contain carry 2193:2197 arg2 oligosaccharide 2219:2233 the complex capping oligosaccharide human natural killer-1 2199:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 12 42 contain carry 2193:2197 arg1 KARs 2188:2191 native KARs 2181:2191 native KARs 2181:2191 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 12 43 theme capping 2211:2217 arg1 oligosaccharide 2219:2233 the complex capping oligosaccharide human natural killer-1 2199:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 13 44 theme KAR 2445:2447 arg1 function 2449:2456 KAR function 2445:2456 KAR function 2445:2456 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 6 45 theme receptors 1149:1157 arg1 function 1131:1138 the function 1127:1138 the function of these receptors 1127:1157 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 7 46 dep proteins 1225:1232 arg1 with 1234:1237 with 1234:1237 with 1234:1237 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 8 47 gly N-glycosylation 1430:1444 arg1 iGluRs 1480:1485 iGluRs 1480:1485 iGluRs 1480:1485 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 13 48 theme potential 2389:2397 arg1 mechanism 2399:2407 a potential mechanism 2387:2407 a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure 2387:2494 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 11 49 attach attached 2151:2158 arg1 identity 2052:2059 the identity 2048:2059 the identity of both the modified sugars 2048:2087 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 49 attach attached 2151:2158 arg1 composition 2105:2115 the subunit composition 2093:2115 the subunit composition of the receptor 2093:2131 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 49 attach attached 2151:2158 arg2 they 2142:2145 they 2142:2145 they 2142:2145 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 8 50 theme iGluRs 1480:1485 arg1 folding 1469:1475 proper folding 1462:1475 proper folding of iGluRs in mammalian cells 1462:1504 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 10 51 theme iGluR 1790:1794 arg1 subunits 1796:1803 iGluR subunits 1790:1803 iGluR subunits that position them for involvement in the structural changes underlying gating 1790:1882 Additionally, they are attached at sites along iGluR subunits that position them for involvement in the structural changes underlying gating. 28714086 1 52 theme hybrid 234:239 arg1 oligosaccharides 253:268 high-mannose, hybrid and complex oligosaccharides 220:268 high-mannose, hybrid and complex oligosaccharides 220:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 13 53 theme function 2449:2456 arg1 fine-tuning 2430:2440 context-specific fine-tuning 2413:2440 context-specific fine-tuning of KAR function through diversity in glycan structure 2413:2494 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 9 54 theme many 1678:1681 arg1 interactions 1699:1710 many protein-protein interactions 1678:1710 many protein-protein interactions 1678:1710 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 1 55 theme complex 245:251 arg1 oligosaccharides 253:268 high-mannose, hybrid and complex oligosaccharides 220:268 high-mannose, hybrid and complex oligosaccharides 220:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 2 56 theme receptor 302:309 arg1 biogenesis 311:320 receptor biogenesis 302:320 receptor biogenesis 302:320 N-glycosylation is crucial for receptor biogenesis, influences receptor trafficking and provides a binding site for carbohydrate binding proteins. 28714086 4 57 theme glycan 610:615 arg1 content 617:623 glycan content 610:623 glycan content 610:623 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 11 58 theme dependent 2035:2043 arg1 manner 2028:2033 a manner 2026:2033 a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached 2026:2158 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 2 59 theme receptor 334:341 arg1 trafficking 343:353 receptor trafficking 334:353 receptor trafficking 334:353 N-glycosylation is crucial for receptor biogenesis, influences receptor trafficking and provides a binding site for carbohydrate binding proteins. 28714086 5 60 theme native 883:888 arg1 KARs 890:893 native KARs 883:893 native KARs 883:893 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 4 61 theme iGluRs 669:674 arg1 receptors 636:644 kainate receptors 628:644 kainate receptors (KARs) 628:651 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 61 theme iGluRs 669:674 arg1 subfamily 656:664 a subfamily 654:664 a subfamily of iGluRs 654:674 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 1 62 theme high-mannose 220:231 arg1 oligosaccharides 253:268 high-mannose, hybrid and complex oligosaccharides 220:268 high-mannose, hybrid and complex oligosaccharides 220:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 3 63 theme structural 563:572 arg1 changes 574:580 the structural changes 559:580 the structural changes underlying gating 559:598 Glycan moieties are large, polar and occasionally charged, and they are attached at sites along iGluRs that position them for involvement in the structural changes underlying gating. 28714086 1 64 theme mature 190:195 arg1 receptors 197:205 mature receptors 190:205 mature receptors that contain high-mannose, hybrid and complex oligosaccharides 190:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 11 65 theme subunit 2097:2103 arg1 composition 2105:2115 the subunit composition 2093:2115 the subunit composition of the receptor 2093:2131 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 8 66 theme mammalian 1490:1498 arg1 cells 1500:1504 mammalian cells 1490:1504 mammalian cells 1490:1504 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 1 67 gly N-glycosylated 130:143 arg2 sites 153:157 4-12 sites 148:157 4-12 sites 148:157 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 67 gly N-glycosylated 130:143 arg1 subunits 117:124 Ionotropic glutamate receptor (iGluR) subunits 79:124 Ionotropic glutamate receptor (iGluR) subunits 79:124 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 67 gly N-glycosylated 130:143 arg1 POINTS 71:76 KEY POINTS 67:76 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 67:269 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 11 68 theme present 1892:1898 arg1 study 1900:1904 the present study 1888:1904 the present study 1888:1904 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 69 theme functional 1984:1993 arg1 properties 1995:2004 the functional properties 1980:2004 the functional properties of the receptors 1980:2021 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 70 theme receptor 2124:2131 arg1 identity 2052:2059 the identity 2048:2059 the identity of both the modified sugars 2048:2087 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 11 70 theme receptor 2124:2131 arg1 composition 2105:2115 the subunit composition 2093:2115 the subunit composition of the receptor 2093:2131 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 7 71 theme post-translational 1397:1414 arg1 modification 1416:1427 this post-translational modification 1392:1427 this post-translational modification 1392:1427 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 13 72 theme glycan 2479:2484 arg1 structure 2486:2494 glycan structure 2479:2494 glycan structure 2479:2494 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 1 73 dep POINTS 71:76 arg1 produces 181:188 produces 181:188 produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides 181:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 73 dep POINTS 71:76 arg1 N-glycosylated 130:143 N-glycosylated 130:143 N-glycosylated 130:143 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 11 74 theme receptors 2013:2021 arg1 properties 1995:2004 the functional properties 1980:2004 the functional properties of the receptors 1980:2021 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 5 75 theme charged 838:844 arg1 HNK-1 860:864 the charged trisaccharide HNK-1 834:864 the charged trisaccharide HNK-1 834:864 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 6 76 theme N-glycans 1027:1035 arg1 composition 1012:1022 the molecular composition 998:1022 the molecular composition of N-glycans 998:1035 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 7 77 theme diversity 1369:1377 arg1 degree 1348:1353 a high degree 1341:1353 a high degree of structural diversity conferred by this post-translational modification 1341:1427 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 9 78 dep large 1631:1635 arg1 polar 1638:1642 polar 1638:1642 polar 1638:1642 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 9 78 dep large 1631:1635 arg1 charged 1658:1664 charged 1658:1664 charged 1658:1664 Glycan moieties are large, polar, occasionally charged and mediate many protein-protein interactions throughout the nervous system. 28714086 13 79 theme cell 2306:2309 arg1 types 2311:2315 cell types 2306:2315 cell types 2306:2315 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 2 80 theme carbohydrate 387:398 arg1 proteins 408:415 carbohydrate binding proteins 387:415 carbohydrate binding proteins 387:415 N-glycosylation is crucial for receptor biogenesis, influences receptor trafficking and provides a binding site for carbohydrate binding proteins. 28714086 1 81 theme KEY 67:69 arg1 POINTS 71:76 KEY POINTS 67:76 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 67:269 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 8 82 theme receptors 1586:1594 arg1 function 1554:1561 the function 1550:1561 the function of successfully folded receptors 1550:1594 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 5 83 theme first 811:815 arg1 observation 817:827 the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties 807:973 the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties 807:973 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 1 84 theme glutamate 90:98 arg1 receptor 100:107 Ionotropic glutamate receptor 79:107 Ionotropic glutamate receptor (iGluR) subunits 79:124 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 84 theme glutamate 90:98 arg1 iGluR 110:114 iGluR 110:114 iGluR 110:114 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 3 85 theme Glycan 418:423 arg1 moieties 425:432 Glycan moieties 418:432 Glycan moieties 418:432 Glycan moieties are large, polar and occasionally charged, and they are attached at sites along iGluRs that position them for involvement in the structural changes underlying gating. 28714086 2 86 theme binding 400:406 arg1 proteins 408:415 carbohydrate binding proteins 387:415 carbohydrate binding proteins 387:415 N-glycosylation is crucial for receptor biogenesis, influences receptor trafficking and provides a binding site for carbohydrate binding proteins. 28714086 0 87 theme kainate 27:33 arg1 properties 55:64 kainate receptor functional properties 27:64 kainate receptor functional properties 27:64 N-glycan content modulates kainate receptor functional properties. 28714086 13 88 theme Glycosylation 2259:2271 arg1 patterns 2273:2280 Glycosylation patterns 2259:2280 Glycosylation patterns 2259:2280 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 5 89 theme functional 953:962 arg1 properties 964:973 recombinant KAR functional properties 937:973 recombinant KAR functional properties 937:973 We report the first observation that the charged trisaccharide HNK-1 is conjugated to native KARs, and we find that it substantially alters recombinant KAR functional properties. 28714086 12 90 theme natural 2241:2247 arg1 killer-1 2249:2256 human natural killer-1 2235:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 11 91 theme glycan 1929:1934 arg1 content 1936:1942 glycan content 1929:1942 glycan content 1929:1942 In the present study, we show that altering glycan content on kainate receptors (KARs) changes the functional properties of the receptors in a manner dependent on the identity of both the modified sugars and the subunit composition of the receptor to which they are attached. 28714086 7 92 theme Ionotropic 1170:1179 arg1 proteins 1225:1232 tetrameric proteins 1214:1232 tetrameric proteins 1214:1232 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 92 theme Ionotropic 1170:1179 arg1 ABSTRACT 1160:1167 ABSTRACT 1160:1167 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 1160:1428 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 92 theme Ionotropic 1170:1179 arg1 iGluRs 1202:1207 iGluRs 1202:1207 iGluRs 1202:1207 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 7 92 theme Ionotropic 1170:1179 arg1 receptors 1191:1199 Ionotropic glutamate receptors 1170:1199 Ionotropic glutamate receptors (iGluRs) 1170:1208 ABSTRACT: Ionotropic glutamate receptors (iGluRs) are tetrameric proteins with between four and 12 consensus sites for N-glycosylation on each subunit, which potentially allows for a high degree of structural diversity conferred by this post-translational modification. 28714086 13 93 theme context-specific 2413:2428 arg1 fine-tuning 2430:2440 context-specific fine-tuning 2413:2440 context-specific fine-tuning of KAR function through diversity in glycan structure 2413:2494 Glycosylation patterns probably differ between cell types, across development or with pathologies, and thus our findings reveal a potential mechanism for context-specific fine-tuning of KAR function through diversity in glycan structure. 28714086 6 94 theme biophysical 1055:1065 arg1 properties 1067:1076 KAR biophysical properties 1051:1076 KAR biophysical properties 1051:1076 Our results show that the molecular composition of N-glycans can influence KAR biophysical properties, revealing a potential mechanism for fine-tuning the function of these receptors. 28714086 10 95 attach attached 1766:1773 arg2 they 1757:1760 they 1757:1760 they 1757:1760 Additionally, they are attached at sites along iGluR subunits that position them for involvement in the structural changes underlying gating. 28714086 10 95 attach attached 1766:1773 arg3 sites 1778:1782 sites 1778:1782 sites along iGluR subunits that position them for involvement in the structural changes underlying gating 1778:1882 Additionally, they are attached at sites along iGluR subunits that position them for involvement in the structural changes underlying gating. 28714086 1 96 contain contain 212:218 arg1 receptors 197:205 mature receptors 190:205 mature receptors that contain high-mannose, hybrid and complex oligosaccharides 190:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 1 96 contain contain 212:218 arg2 oligosaccharides 253:268 high-mannose, hybrid and complex oligosaccharides 220:268 high-mannose, hybrid and complex oligosaccharides 220:268 KEY POINTS: Ionotropic glutamate receptor (iGluR) subunits are N-glycosylated at 4-12 sites, and Golgi processing produces mature receptors that contain high-mannose, hybrid and complex oligosaccharides. 28714086 12 97 theme complex 2203:2209 arg1 oligosaccharide 2219:2233 the complex capping oligosaccharide human natural killer-1 2199:2256 the complex capping oligosaccharide human natural killer-1 2199:2256 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 28714086 4 98 theme kainate 628:634 arg1 KARs 647:650 KARs 647:650 KARs 647:650 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 98 theme kainate 628:634 arg1 receptors 636:644 kainate receptors 628:644 kainate receptors (KARs) 628:651 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 4 98 theme kainate 628:634 arg1 subfamily 656:664 a subfamily 654:664 a subfamily of iGluRs 654:674 Altering glycan content on kainate receptors (KARs), a subfamily of iGluRs, changes functional properties of the receptor, such as desensitization, recovery from desensitization and deactivation. 28714086 8 99 theme proper 1462:1467 arg1 folding 1469:1475 proper folding 1462:1475 proper folding of iGluRs in mammalian cells 1462:1504 N-glycosylation is required for proper folding of iGluRs in mammalian cells, although the impact of oligosaccharides on the function of successfully folded receptors is less clear. 28714086 12 100 theme native 2181:2186 arg1 KARs 2188:2191 native KARs 2181:2191 native KARs 2181:2191 We also report that native KARs carry the complex capping oligosaccharide human natural killer-1. 29274340 8 0 theme ions 1711:1714 arg1 types 1928:1932 two types 1924:1932 two types 1924:1932 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 0 theme ions 1711:1714 arg1 chromatograms 1674:1686 the extracted-ion chromatograms 1656:1686 the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types) 1656:1933 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 0 theme ions 1711:1714 arg1 nano-LC-ESI-HCD-MS/MS 1629:1649 nano-LC-ESI-HCD-MS/MS 1629:1649 nano-LC-ESI-HCD-MS/MS 1629:1649 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 1 1 theme high 404:407 arg1 dose 409:412 a high dose 402:412 a high dose 402:412 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 2 2 theme replacement 558:568 arg1 therapy 570:576 enzyme replacement therapy 551:576 enzyme replacement therapy (ERT) 551:582 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 2 theme replacement 558:568 arg1 ERT 579:581 ERT 579:581 ERT 579:581 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 1 3 theme human 264:268 arg1 drug 328:331 the only drug 319:331 the only drug approved for treating Pompe disease 319:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 3 theme human 264:268 arg1 rhGAA 294:298 rhGAA 294:298 rhGAA 294:298 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 3 theme human 264:268 arg1 required 417:424 required 417:424 is required 414:424 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 3 theme human 264:268 arg1 alpha-glucosidase 275:291 a recombinant human acid alpha-glucosidase 250:291 a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease 250:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 3 theme human 264:268 arg1 Myozyme 239:245 Myozyme 239:245 Myozyme 239:245 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 8 4 theme glycosylation 1736:1748 arg1 sites 1750:1754 three M6P glycosylation sites 1726:1754 three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1726:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 6 5 theme M6P 1320:1322 arg1 glycans 1324:1330 hybrid-type M6P glycans 1308:1330 hybrid-type M6P glycans (2 variants, 11.2%) 1308:1350 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 0 6 theme found 125:129 arg1 233 135:137 newly found Asn 233 119:137 newly found Asn 233 119:137 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 6 7 theme hybrid-type 1357:1367 arg1 glycans 1373:1379 hybrid-type M6P glycans 1357:1379 hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%) 1357:1417 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 2 8 theme lysosomal 504:512 arg1 targeting 521:529 lysosomal enzyme targeting 504:529 lysosomal enzyme targeting 504:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 3 9 theme liquid 737:742 arg1 LC 760:761 LC 760:761 LC 760:761 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 9 theme liquid 737:742 arg1 chromatography 744:757 liquid chromatography 737:757 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 6 10 theme high-mannose-type 1216:1232 arg1 glycans 1238:1244 high-mannose-type M6P glycans 1216:1244 high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%) 1216:1305 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 7 11 link M6P-derived 1527:1537 arg1 ions 1547:1550 six distinctive M6P-derived oxonium ions 1511:1550 six distinctive M6P-derived oxonium ions 1511:1550 HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. 29274340 1 12 theme efficacy 382:389 arg1 means 391:395 its low efficacy means 374:395 its low efficacy means that a high dose 374:412 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 3 13 theme dissociation 818:829 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 13 theme dissociation 818:829 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 9 14 theme rhGAA 1977:1981 arg1 This 1950:1953 This 1950:1953 This 1950:1953 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 9 14 theme rhGAA 1977:1981 arg1 study 1968:1972 the first study 1958:1972 the first study of rhGAA to differentiate M6P glycans and identify their attachment sites 1958:2046 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 6 15 theme found 1193:1197 arg1 ones 1199:1202 3 newly found ones 1185:1202 4 types: 3 newly found ones 1176:1202 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 5 16 theme relative 1006:1013 arg1 % 1027:1027 % 1027:1027 % 1027:1027 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 16 theme relative 1006:1013 arg1 quantities 1015:1024 The relative quantities 1002:1024 The relative quantities (%) of 78 glycans 1002:1042 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 3 17 theme -high-energy 793:804 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 17 theme -high-energy 793:804 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 7 18 theme M6P-derived 1527:1537 arg1 ions 1547:1550 six distinctive M6P-derived oxonium ions 1511:1550 six distinctive M6P-derived oxonium ions 1511:1550 HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. 29274340 0 19 theme human 155:159 arg1 alpha-glucosidase 166:182 recombinant human acid alpha-glucosidase 143:182 recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease 143:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 19 theme human 155:159 arg1 treatment 210:218 the only approved treatment 192:218 the only approved treatment for Pompe disease 192:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 7 20 theme HCD-MS/MS 1482:1490 arg1 spectra 1492:1498 HCD-MS/MS spectra 1482:1498 HCD-MS/MS spectra 1482:1498 HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. 29274340 3 21 theme ESI 789:791 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 21 theme ESI 789:791 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 8 22 theme Asn 1803:1805 arg1 470 1807:1809 Asn 470 1803:1809 Asn 470 1803:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 3 23 theme HCD 832:834 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 23 theme HCD 832:834 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 2 24 theme complex 598:604 arg1 structure 606:614 its complex structure 594:614 its complex structure 594:614 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 3 25 theme ionization 777:786 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 25 theme ionization 777:786 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 8 26 theme extracted-ion 1660:1672 arg1 types 1928:1932 two types 1924:1932 two types 1924:1932 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 26 theme extracted-ion 1660:1672 arg1 chromatograms 1674:1686 the extracted-ion chromatograms 1656:1686 the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types) 1656:1933 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 3 27 gly glycosylation 708:720 arg1 rhGAA 725:729 rhGAA 725:729 rhGAA 725:729 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 9 28 theme attachment 2031:2040 arg1 sites 2042:2046 their attachment sites 2025:2046 their attachment sites 2025:2046 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 1 29 theme Pompe 355:359 arg1 disease 361:367 Pompe disease 355:367 Pompe disease 355:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 2 30 theme M6P 448:450 arg1 glycosylation 453:465 Mannose-6-phosphate (M6P) glycosylation 427:465 Mannose-6-phosphate (M6P) glycosylation on rhGAA 427:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 30 theme M6P 448:450 arg1 factor 485:490 a key factor 479:490 a key factor influencing lysosomal enzyme targeting 479:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 6 31 theme M6P 1449:1451 arg1 glycans 1453:1459 high-mannose-type M6P glycans 1431:1459 high-mannose-type M6P glycans (3 variants, 64.4%) 1431:1479 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 0 32 dep sites 102:106 arg1 including 109:117 including 109:117 including newly found Asn 233 109:137 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 8 33 theme M6P 1875:1877 arg1 glycans 1879:1885 eight M6P glycans 1869:1885 eight M6P glycans (four types) 1869:1898 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 33 theme M6P 1875:1877 arg1 types 1893:1897 four types 1888:1897 four types 1888:1897 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 2 34 theme small 631:635 arg1 quantity 637:644 relatively small quantity 620:644 relatively small quantity 620:644 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 8 35 attach attached 1811:1818 arg1 glycans 1879:1885 eight M6P glycans 1869:1885 eight M6P glycans (four types) 1869:1898 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg2 types 1928:1932 two types 1924:1932 two types 1924:1932 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg1 types 1861:1865 two types 1857:1865 two types 1857:1865 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg2 chromatograms 1674:1686 the extracted-ion chromatograms 1656:1686 the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types) 1656:1933 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg1 glycans 1915:1921 seven M6P glycans 1905:1921 seven M6P glycans 1905:1921 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg1 types 1893:1897 four types 1888:1897 four types 1888:1897 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 35 attach attached 1811:1818 arg1 glycans 1848:1854 nine M6P glycans 1839:1854 nine M6P glycans (two types) 1839:1866 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 36 theme M6P 1844:1846 arg1 glycans 1848:1854 nine M6P glycans 1839:1854 nine M6P glycans (two types) 1839:1866 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 36 theme M6P 1844:1846 arg1 types 1861:1865 two types 1857:1865 two types 1857:1865 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 1 37 theme acid 270:273 arg1 drug 328:331 the only drug 319:331 the only drug approved for treating Pompe disease 319:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 37 theme acid 270:273 arg1 rhGAA 294:298 rhGAA 294:298 rhGAA 294:298 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 37 theme acid 270:273 arg1 required 417:424 required 417:424 is required 414:424 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 37 theme acid 270:273 arg1 alpha-glucosidase 275:291 a recombinant human acid alpha-glucosidase 250:291 a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease 250:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 37 theme acid 270:273 arg1 Myozyme 239:245 Myozyme 239:245 Myozyme 239:245 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 8 38 theme oxonium 1703:1709 arg1 ions 1711:1714 M6P-derived oxonium ions 1691:1714 M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1691:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 4 39 theme MS/MS 995:999 arg1 efficiency 961:970 mass ionization efficiency 945:970 mass ionization efficiency 945:970 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 4 39 theme MS/MS 995:999 arg1 sensitivity 980:990 the sensitivity 976:990 the sensitivity of MS/MS 976:999 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 2 40 gly glycosylation 453:465 arg1 rhGAA 470:474 rhGAA 470:474 rhGAA 470:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 4 41 theme ionization 950:959 arg1 efficiency 961:970 mass ionization efficiency 945:970 mass ionization efficiency 945:970 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 0 42 theme glycans 25:31 arg1 types 16:20 Four unreported types 0:20 Four unreported types of glycans containing mannose-6-phosphate 0:62 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 8 43 dep Asn 1776:1778 arg1 found 1791:1795 found 1791:1795 found 1791:1795 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 2 44 theme Mannose-6-phosphate 427:445 arg1 glycosylation 453:465 Mannose-6-phosphate (M6P) glycosylation 427:465 Mannose-6-phosphate (M6P) glycosylation on rhGAA 427:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 44 theme Mannose-6-phosphate 427:445 arg1 factor 485:490 a key factor 479:490 a key factor influencing lysosomal enzyme targeting 479:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 9 45 theme approved 2080:2087 arg1 drug 2089:2092 an approved drug 2077:2092 an approved drug for Pompe disease 2077:2110 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 9 45 theme approved 2080:2087 arg1 rhGAA 2057:2061 rhGAA 2057:2061 rhGAA 2057:2061 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 7 46 theme oxonium 1539:1545 arg1 ions 1547:1550 six distinctive M6P-derived oxonium ions 1511:1550 six distinctive M6P-derived oxonium ions 1511:1550 HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. 29274340 3 47 theme tandem 837:842 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 47 theme tandem 837:842 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 8 48 gly glycosylation 1736:1748 arg2 sites 1750:1754 three M6P glycosylation sites 1726:1754 three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1726:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 48 gly glycosylation 1736:1748 arg2 three 1726:1730 three 1726:1730 three 1726:1730 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 2 49 theme therapy 570:576 arg1 efficacy 539:546 the efficacy 535:546 the efficacy of enzyme replacement therapy (ERT) 535:582 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 49 theme therapy 570:576 arg1 glycosylation 453:465 Mannose-6-phosphate (M6P) glycosylation 427:465 Mannose-6-phosphate (M6P) glycosylation on rhGAA 427:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 49 theme therapy 570:576 arg1 factor 485:490 a key factor 479:490 a key factor influencing lysosomal enzyme targeting 479:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 8 50 theme M6P 1732:1734 arg1 sites 1750:1754 three M6P glycosylation sites 1726:1754 three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1726:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 1 51 theme recombinant 252:262 arg1 drug 328:331 the only drug 319:331 the only drug approved for treating Pompe disease 319:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 51 theme recombinant 252:262 arg1 rhGAA 294:298 rhGAA 294:298 rhGAA 294:298 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 51 theme recombinant 252:262 arg1 required 417:424 required 417:424 is required 414:424 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 51 theme recombinant 252:262 arg1 alpha-glucosidase 275:291 a recombinant human acid alpha-glucosidase 250:291 a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease 250:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 51 theme recombinant 252:262 arg1 Myozyme 239:245 Myozyme 239:245 Myozyme 239:245 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 6 52 dep types 1178:1182 arg1 ones 1199:1202 3 newly found ones 1185:1202 4 types: 3 newly found ones 1176:1202 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 2 53 theme enzyme 551:556 arg1 therapy 570:576 enzyme replacement therapy 551:576 enzyme replacement therapy (ERT) 551:582 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 53 theme enzyme 551:556 arg1 ERT 579:581 ERT 579:581 ERT 579:581 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 6 54 dep glycans 1373:1379 arg1 variants 1403:1410 3 variants 1401:1410 3 variants 1401:1410 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 6 54 dep glycans 1373:1379 arg1 % 1416:1416 6.9% 1413:1416 6.9% 1413:1416 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 0 55 theme approved 201:208 arg1 alpha-glucosidase 166:182 recombinant human acid alpha-glucosidase 143:182 recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease 143:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 55 theme approved 201:208 arg1 treatment 210:218 the only approved treatment 192:218 the only approved treatment for Pompe disease 192:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 56 contain containing 33:42 arg2 mannose-6-phosphate 44:62 mannose-6-phosphate 44:62 mannose-6-phosphate 44:62 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 56 contain containing 33:42 arg1 glycans 25:31 glycans 25:31 glycans containing mannose-6-phosphate 25:62 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 57 theme Asn 131:133 arg1 233 135:137 newly found Asn 233 119:137 newly found Asn 233 119:137 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 6 58 theme M6P 1369:1371 arg1 glycans 1373:1379 hybrid-type M6P glycans 1357:1379 hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%) 1357:1417 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 2 59 theme enzyme 514:519 arg1 targeting 521:529 lysosomal enzyme targeting 504:529 lysosomal enzyme targeting 504:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 60 from glycosylation 453:465 arg1 rhGAA 470:474 rhGAA 470:474 rhGAA 470:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 3 61 theme M6P 704:706 arg1 glycosylation 708:720 M6P glycosylation 704:720 M6P glycosylation 704:720 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 6 62 theme M6P 1234:1236 arg1 glycans 1238:1244 high-mannose-type M6P glycans 1216:1244 high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%) 1216:1305 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 8 63 link M6P-derived 1691:1701 arg1 ions 1711:1714 M6P-derived oxonium ions 1691:1714 M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1691:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 5 64 contain containing 1089:1098 arg2 glycans 1109:1115 M6P glycans 1105:1115 M6P glycans 1105:1115 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 64 contain containing 1089:1098 arg2 M6P 1100:1102 M6P 1100:1102 M6P (M6P glycans) 1100:1116 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 64 contain containing 1089:1098 arg1 glycans 1081:1087 glycans 1081:1087 glycans containing M6P (M6P glycans) 1081:1116 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 3 65 theme chromatography 744:757 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 65 theme chromatography 744:757 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 0 66 theme recombinant 143:153 arg1 alpha-glucosidase 166:182 recombinant human acid alpha-glucosidase 143:182 recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease 143:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 66 theme recombinant 143:153 arg1 treatment 210:218 the only approved treatment 192:218 the only approved treatment for Pompe disease 192:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 1 67 theme low 378:380 arg1 means 391:395 its low efficacy means 374:395 its low efficacy means that a high dose 374:412 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 6 68 dep glycans 1238:1244 arg1 % 1304:1304 17.5% 1300:1304 17.5% 1300:1304 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 6 68 dep glycans 1238:1244 arg1 variants 1290:1297 2 variants 1288:1297 2 variants 1288:1297 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 0 69 theme acid 161:164 arg1 alpha-glucosidase 166:182 recombinant human acid alpha-glucosidase 143:182 recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease 143:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 69 theme acid 161:164 arg1 treatment 210:218 the only approved treatment 192:218 the only approved treatment for Pompe disease 192:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 8 70 theme protease-digested 1585:1601 arg1 rhGAA 1603:1607 protease-digested rhGAA 1585:1607 protease-digested rhGAA 1585:1607 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 8 71 gly glycopeptides 1557:1569 arg2 glycopeptides 1557:1569 The glycopeptides 1553:1569 The glycopeptides obtained from protease-digested rhGAA 1553:1607 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 3 72 theme -electrospray 763:775 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 72 theme -electrospray 763:775 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 73 theme collisional 806:816 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 73 theme collisional 806:816 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 9 74 theme M6P 2000:2002 arg1 glycans 2004:2010 M6P glycans 2000:2010 M6P glycans 2000:2010 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 1 75 theme only 323:326 arg1 drug 328:331 the only drug 319:331 the only drug approved for treating Pompe disease 319:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 75 theme only 323:326 arg1 required 417:424 required 417:424 is required 414:424 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 75 theme only 323:326 arg1 alpha-glucosidase 275:291 a recombinant human acid alpha-glucosidase 250:291 a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease 250:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 75 theme only 323:326 arg1 Myozyme 239:245 Myozyme 239:245 Myozyme 239:245 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 3 76 theme mass 844:847 arg1 MS/MS 863:867 MS/MS 863:867 MS/MS 863:867 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 3 76 theme mass 844:847 arg1 spectrometry 849:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry 737:860 liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) 737:868 This study investigated M6P glycosylation on rhGAA using liquid chromatography (LC)-electrospray ionization (ESI)-high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS). 29274340 1 77 theme approved 333:340 arg1 drug 328:331 the only drug 319:331 the only drug approved for treating Pompe disease 319:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 77 theme approved 333:340 arg1 required 417:424 required 417:424 is required 414:424 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 77 theme approved 333:340 arg1 alpha-glucosidase 275:291 a recombinant human acid alpha-glucosidase 250:291 a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease 250:367 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 1 77 theme approved 333:340 arg1 Myozyme 239:245 Myozyme 239:245 Myozyme 239:245 Myozyme is a recombinant human acid alpha-glucosidase (rhGAA) that is currently the only drug approved for treating Pompe disease, and its low efficacy means that a high dose is required. 29274340 0 78 theme Pompe 224:228 arg1 disease 230:236 Pompe disease 224:236 Pompe disease 224:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 7 79 theme distinctive 1515:1525 arg1 ions 1547:1550 six distinctive M6P-derived oxonium ions 1511:1550 six distinctive M6P-derived oxonium ions 1511:1550 HCD-MS/MS spectra identified six distinctive M6P-derived oxonium ions. 29274340 2 80 theme key 481:483 arg1 glycosylation 453:465 Mannose-6-phosphate (M6P) glycosylation 427:465 Mannose-6-phosphate (M6P) glycosylation on rhGAA 427:474 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 2 80 theme key 481:483 arg1 factor 485:490 a key factor 479:490 a key factor influencing lysosomal enzyme targeting 479:529 Mannose-6-phosphate (M6P) glycosylation on rhGAA is a key factor influencing lysosomal enzyme targeting and the efficacy of enzyme replacement therapy (ERT); however, its complex structure and relatively small quantity still remain to be characterized. 29274340 5 81 theme glycans 1036:1042 arg1 % 1027:1027 % 1027:1027 % 1027:1027 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 81 theme glycans 1036:1042 arg1 quantities 1015:1024 The relative quantities 1002:1024 The relative quantities (%) of 78 glycans 1002:1042 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 0 82 theme unreported 5:14 arg1 types 16:20 Four unreported types 0:20 Four unreported types of glycans containing mannose-6-phosphate 0:62 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 6 83 theme high-mannose-type 1431:1447 arg1 glycans 1453:1459 high-mannose-type M6P glycans 1431:1459 high-mannose-type M6P glycans (3 variants, 64.4%) 1431:1479 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 4 84 attach released 883:890 arg2 glycans 875:881 The glycans 871:881 The glycans released from rhGAA 871:901 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 4 84 attach released 883:890 arg1 rhGAA 897:901 rhGAA 897:901 rhGAA 897:901 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 6 85 theme hybrid-type 1308:1318 arg1 glycans 1324:1330 hybrid-type M6P glycans 1308:1330 hybrid-type M6P glycans (2 variants, 11.2%) 1308:1350 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 8 86 theme M6P-derived 1691:1701 arg1 ions 1711:1714 M6P-derived oxonium ions 1691:1714 M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 1691:1809 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 6 87 dep glycans 1453:1459 arg1 % 1478:1478 64.4% 1474:1478 64.4% 1474:1478 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 6 87 dep glycans 1453:1459 arg1 variants 1464:1471 3 variants 1462:1471 3 variants 1462:1471 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 5 88 theme M6P 1105:1107 arg1 M6P 1100:1102 M6P 1100:1102 M6P (M6P glycans) 1100:1116 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 88 theme M6P 1105:1107 arg1 glycans 1109:1115 M6P glycans 1105:1115 M6P glycans 1105:1115 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 4 89 theme mass 945:948 arg1 efficiency 961:970 mass ionization efficiency 945:970 mass ionization efficiency 945:970 The glycans released from rhGAA were labeled with procainamide to improve mass ionization efficiency and the sensitivity of MS/MS. 29274340 5 90 theme them 1071:1074 arg1 them 1071:1074 them 1071:1074 them 1071:1074 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 5 90 theme them 1071:1074 arg1 % 1066:1066 1.0% 1063:1066 1.0% of them 1063:1074 The relative quantities (%) of 78 glycans were obtained, and 1.0% of them were glycans containing M6P (M6P glycans). 29274340 0 91 attach attached 84:91 arg1 treatment 210:218 the only approved treatment 192:218 the only approved treatment for Pompe disease 192:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 91 attach attached 84:91 arg1 alpha-glucosidase 166:182 recombinant human acid alpha-glucosidase 143:182 recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease 143:236 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 91 attach attached 84:91 arg3 sites 102:106 three sites 96:106 three sites (including newly found Asn 233) 96:138 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 0 91 attach attached 84:91 arg2 types 16:20 Four unreported types 0:20 Four unreported types of glycans containing mannose-6-phosphate 0:62 Four unreported types of glycans containing mannose-6-phosphate are heterogeneously attached at three sites (including newly found Asn 233) to recombinant human acid alpha-glucosidase that is the only approved treatment for Pompe disease. 29274340 8 92 theme M6P 1911:1913 arg1 glycans 1915:1921 seven M6P glycans 1905:1921 seven M6P glycans 1905:1921 The glycopeptides obtained from protease-digested rhGAA were analyzed using nano-LC-ESI-HCD-MS/MS, and the extracted-ion chromatograms of M6P-derived oxonium ions confirmed three M6P glycosylation sites comprising Asn 140, Asn 233 (newly found), and Asn 470 attached heterogeneously to nine M6P glycans (two types), eight M6P glycans (four types), and seven M6P glycans (two types), respectively. 29274340 6 93 dep glycans 1324:1330 arg1 variants 1335:1342 2 variants 1333:1342 2 variants 1333:1342 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 6 93 dep glycans 1324:1330 arg1 % 1349:1349 11.2% 1345:1349 11.2% 1345:1349 These were categorized according to their structure into 4 types: 3 newly found ones, comprising high-mannose-type M6P glycans capped with N-acetylglucosamine (GlcNAc) (2 variants, 17.5%), hybrid-type M6P glycans (2 variants, 11.2%), and hybrid-type M6P glycans capped with GlcNAc (3 variants, 6.9%), as well as high-mannose-type M6P glycans (3 variants, 64.4%). 29274340 9 94 theme Pompe 2098:2102 arg1 disease 2104:2110 Pompe disease 2098:2110 Pompe disease 2098:2110 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 9 95 theme first 1962:1966 arg1 This 1950:1953 This 1950:1953 This 1950:1953 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 29274340 9 95 theme first 1962:1966 arg1 study 1968:1972 the first study 1958:1972 the first study of rhGAA to differentiate M6P glycans and identify their attachment sites 1958:2046 This is the first study of rhGAA to differentiate M6P glycans and identify their attachment sites, despite rhGAA already being an approved drug for Pompe disease. 28489325 1 0 theme leukocytes 231:240 arg1 surface 215:221 the surface 211:221 the surface of most leukocytes 211:240 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 4 1 theme binding 926:932 arg1 LE010 961:965 a symmetry-related LE010 942:965 a symmetry-related LE010 942:965 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 4 1 theme binding 926:932 arg1 site 934:937 the binding site 922:937 the binding site of a symmetry-related LE010 922:965 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 3 2 attach attached 778:785 arg2 N22Q 727:730 N22Q 727:730 N22Q 727:730 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 2 attach attached 778:785 arg2 N139Q 736:740 N139Q 736:740 N139Q 736:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 2 attach attached 778:785 arg1 Asn66 790:794 Asn66 790:794 Asn66 790:794 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 2 attach attached 778:785 arg2 exchanges 717:725 exchanges N22Q and N139Q 717:740 exchanges N22Q and N139Q 717:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 2 3 theme L-selectin 331:340 arg1 structure 312:320 the crystal structure 300:320 the crystal structure of human L-selectin 300:340 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 5 4 theme domain 1011:1016 arg1 similar 1022:1028 similar 1022:1028 similar 1022:1028 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 5 4 theme domain 1011:1016 arg1 orientation 972:982 The orientation 968:982 The orientation of the lectin and EGF-like domain 968:1016 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 2 5 theme glycosylation 503:515 arg1 sites 517:521 the three glycosylation sites 493:521 the three glycosylation sites (Asn22, Asn66 and Asn139) 493:547 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 5 theme glycosylation 503:515 arg1 Asn139 541:546 Asn139 541:546 Asn139 541:546 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 5 theme glycosylation 503:515 arg1 Asn66 531:535 Asn66 531:535 Asn66 531:535 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 5 6 theme E- 1070:1071 arg1 conformation 1054:1065 the described "bent" conformation 1033:1065 the described "bent" conformation of E- and P-selectins 1033:1087 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 4 7 theme terminal 840:847 arg1 mannose 849:855 the terminal mannose 836:855 the terminal mannose of GlcNAc2 Man5 of one LE010 molecule 836:893 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 4 8 theme Man5 868:871 arg1 mannose 849:855 the terminal mannose 836:855 the terminal mannose of GlcNAc2 Man5 of one LE010 molecule 836:893 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 3 9 theme N-glycan 763:770 arg1 chain 772:776 one GlcNAc2 Man5 N-glycan chain 746:776 one GlcNAc2 Man5 N-glycan chain 746:776 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 0 10 theme Lectin 89:94 arg1 L-Selectin 126:135 Human L-Selectin 120:135 Human L-Selectin 120:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 10 theme Lectin 89:94 arg1 Domains 109:115 the Lectin and EGF-Like Domains 85:115 Domains 109:115 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 3 11 dep exchanges 717:725 arg1 N22Q 727:730 N22Q 727:730 N22Q 727:730 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 11 dep exchanges 717:725 arg1 exchanges 717:725 exchanges N22Q and N139Q 717:740 exchanges N22Q and N139Q 717:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 11 dep exchanges 717:725 arg1 N139Q 736:740 N139Q 736:740 N139Q 736:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 2 12 gly glycosylation 503:515 arg2 sites 517:521 the three glycosylation sites 493:521 the three glycosylation sites (Asn22, Asn66 and Asn139) 493:547 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 12 gly glycosylation 503:515 arg2 three 497:501 three 497:501 three 497:501 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 12 gly glycosylation 503:515 arg2 Asn66 531:535 Asn66 531:535 Asn66 531:535 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 12 gly glycosylation 503:515 arg2 Asn139 541:546 Asn139 541:546 Asn139 541:546 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 0 13 theme Domains 109:115 arg1 Structure 72:80 the Crystal Structure 60:80 the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin 60:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 5 14 theme described 1037:1045 arg1 conformation 1054:1065 the described "bent" conformation 1033:1065 the described "bent" conformation of E- and P-selectins 1033:1087 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 6 15 theme -binding 1099:1106 arg1 site 1108:1111 The Ca2+ -binding site 1090:1111 The Ca2+ -binding site 1090:1111 The Ca2+ -binding site reflects the binding mode seen in E- and P-selectin structures co-crystallised with ligands. 28489325 0 16 theme EGF-Like 100:107 arg1 L-Selectin 126:135 Human L-Selectin 120:135 Human L-Selectin 120:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 16 theme EGF-Like 100:107 arg1 Domains 109:115 the Lectin and EGF-Like Domains 85:115 Domains 109:115 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 6 17 theme Ca2+ 1094:1097 arg1 site 1108:1111 The Ca2+ -binding site 1090:1111 The Ca2+ -binding site 1090:1111 The Ca2+ -binding site reflects the binding mode seen in E- and P-selectin structures co-crystallised with ligands. 28489325 5 18 theme bent 1048:1051 arg1 conformation 1054:1065 the described "bent" conformation 1033:1065 the described "bent" conformation of E- and P-selectins 1033:1087 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 2 19 theme shortened 358:366 arg1 version 368:374 a shortened version 356:374 a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) 356:450 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 20 theme EGF-like 423:430 arg1 domains 432:438 the C-type lectin and EGF-like domains 401:438 domains 432:438 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 1 21 theme N-glycosylation 258:272 arg1 sites 274:278 seven N-glycosylation sites 252:278 seven N-glycosylation sites 252:278 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 4 22 theme Crystal-structure 797:813 arg1 analysis 815:822 Crystal-structure analysis 797:822 Crystal-structure analysis 797:822 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 4 23 theme LE010 961:965 arg1 LE010 961:965 a symmetry-related LE010 942:965 a symmetry-related LE010 942:965 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 4 23 theme LE010 961:965 arg1 site 934:937 the binding site 922:937 the binding site of a symmetry-related LE010 922:965 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 6 24 theme binding 1126:1132 arg1 mode 1134:1137 the binding mode 1122:1137 the binding mode seen in E- and P-selectin structures co-crystallised with ligands 1122:1203 The Ca2+ -binding site reflects the binding mode seen in E- and P-selectin structures co-crystallised with ligands. 28489325 0 25 theme Human 120:124 arg1 L-Selectin 126:135 Human L-Selectin 120:135 Human L-Selectin 120:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 2 26 theme C-type 405:410 arg1 lectin 412:417 the C-type lectin and EGF-like domains 401:438 lectin 412:417 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 26 theme C-type 405:410 arg1 LE 448:449 termed LE 441:449 termed LE 441:449 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 3 27 from variant 696:702 arg1 1.9 Å 683:687 1.9 Å 683:687 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66 683:794 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 4 28 theme LE010 880:884 arg1 molecule 886:893 one LE010 molecule 876:893 one LE010 molecule 876:893 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 1 29 theme cell-adhesion 185:197 arg1 L-Selectin 171:180 L-Selectin 171:180 L-Selectin 171:180 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 1 29 theme cell-adhesion 185:197 arg1 receptor 199:206 a cell-adhesion receptor 183:206 a cell-adhesion receptor on the surface of most leukocytes 183:240 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 0 30 theme N-Glycans 42:50 arg1 Microheterogeneity 20:37 Microheterogeneity 20:37 Microheterogeneity 20:37 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 30 theme N-Glycans 42:50 arg1 Macro- 9:14 Macro- 9:14 Macro- 9:14 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 4 31 theme molecule 886:893 arg1 Man5 868:871 GlcNAc2 Man5 860:871 GlcNAc2 Man5 of one LE010 molecule 860:893 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 6 32 theme P-selectin 1154:1163 arg1 structures 1165:1174 E- and P-selectin structures 1147:1174 E- and P-selectin structures co-crystallised with ligands 1147:1203 The Ca2+ -binding site reflects the binding mode seen in E- and P-selectin structures co-crystallised with ligands. 28489325 4 33 theme symmetry-related 944:959 arg1 LE010 961:965 a symmetry-related LE010 942:965 a symmetry-related LE010 942:965 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 2 34 theme sites 517:521 arg1 mutations 480:488 mutations 480:488 mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order 480:556 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 1 35 from receptor 199:206 arg1 surface 215:221 the surface 211:221 the surface of most leukocytes 211:240 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 2 36 theme L-selectin 379:388 arg1 version 368:374 a shortened version 356:374 a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) 356:450 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 37 theme termed 441:446 arg1 lectin 412:417 the C-type lectin and EGF-like domains 401:438 lectin 412:417 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 37 theme termed 441:446 arg1 LE 448:449 termed LE 441:449 termed LE 441:449 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 5 38 theme P-selectins 1077:1087 arg1 conformation 1054:1065 the described "bent" conformation 1033:1065 the described "bent" conformation of E- and P-selectins 1033:1087 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 6 39 theme E- 1147:1148 arg1 structures 1165:1174 E- and P-selectin structures 1147:1174 E- and P-selectin structures co-crystallised with ligands 1147:1203 The Ca2+ -binding site reflects the binding mode seen in E- and P-selectin structures co-crystallised with ligands. 28489325 1 40 gly N-glycosylation 258:272 arg2 seven 252:256 seven 252:256 seven 252:256 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 1 40 gly N-glycosylation 258:272 arg2 sites 274:278 seven N-glycosylation sites 252:278 seven N-glycosylation sites 252:278 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 2 41 theme human 325:329 arg1 L-selectin 331:340 human L-selectin 325:340 human L-selectin 325:340 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 0 42 theme L-Selectin 126:135 arg1 L-Selectin 126:135 Human L-Selectin 120:135 Human L-Selectin 120:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 42 theme L-Selectin 126:135 arg1 Domains 109:115 the Lectin and EGF-Like Domains 85:115 Domains 109:115 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 43 gly Microheterogeneity 20:37 arg1 N-Glycans 42:50 N-Glycans 42:50 N-Glycans 42:50 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 0 44 theme 1.9 Å 153:157 arg1 Resolution 159:168 1.9 Å Resolution 153:168 1.9 Å Resolution 153:168 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 5 45 theme lectin 991:996 arg1 similar 1022:1028 similar 1022:1028 similar 1022:1028 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 5 45 theme lectin 991:996 arg1 orientation 972:982 The orientation 968:982 The orientation of the lectin and EGF-like domain 968:1016 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 3 46 theme GlcNAc2 750:756 arg1 chain 772:776 one GlcNAc2 Man5 N-glycan chain 746:776 one GlcNAc2 Man5 N-glycan chain 746:776 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 4 47 theme GlcNAc2 860:866 arg1 Man5 868:871 GlcNAc2 Man5 860:871 GlcNAc2 Man5 of one LE010 molecule 860:893 Crystal-structure analysis showed that the terminal mannose of GlcNAc2 Man5 of one LE010 molecule was coordinated to Ca2+ in the binding site of a symmetry-related LE010. 28489325 1 48 contain contains 243:250 arg1 L-Selectin 171:180 L-Selectin 171:180 L-Selectin 171:180 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 1 48 contain contains 243:250 arg1 receptor 199:206 a cell-adhesion receptor 183:206 a cell-adhesion receptor on the surface of most leukocytes 183:240 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 1 48 contain contains 243:250 arg2 sites 274:278 seven N-glycosylation sites 252:278 seven N-glycosylation sites 252:278 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 28489325 2 49 theme crystal 304:310 arg1 structure 312:320 the crystal structure 300:320 the crystal structure of human L-selectin 300:340 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 3 50 theme Man5 758:761 arg1 chain 772:776 one GlcNAc2 Man5 N-glycan chain 746:776 one GlcNAc2 Man5 N-glycan chain 746:776 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 5 51 theme EGF-like 1002:1009 arg1 domain 1011:1016 EGF-like domain 1002:1016 EGF-like domain 1002:1016 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 2 52 dep sites 517:521 arg1 sites 517:521 the three glycosylation sites 493:521 the three glycosylation sites (Asn22, Asn66 and Asn139) 493:547 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 52 dep sites 517:521 arg1 Asn139 541:546 Asn139 541:546 Asn139 541:546 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 2 52 dep sites 517:521 arg1 Asn66 531:535 Asn66 531:535 Asn66 531:535 In order to obtain the crystal structure of human L-selectin, we expressed a shortened version of L-selectin comprising the C-type lectin and EGF-like domains (termed LE) and systematically analysed mutations of the three glycosylation sites (Asn22, Asn66 and Asn139) in order to reduce macroheterogeneity. 28489325 3 53 with variant 696:702 arg1 chain 772:776 one GlcNAc2 Man5 N-glycan chain 746:776 one GlcNAc2 Man5 N-glycan chain 746:776 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 53 with variant 696:702 arg1 N139Q 736:740 N139Q 736:740 N139Q 736:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 53 with variant 696:702 arg1 exchanges 717:725 exchanges N22Q and N139Q 717:740 exchanges N22Q and N139Q 717:740 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 3 53 with variant 696:702 arg1 N22Q 727:730 N22Q 727:730 N22Q 727:730 After we further removed microheterogeneity, we obtained crystals that diffracted X-rays up to 1.9 Å from a variant (LE010) with exchanges N22Q and N139Q and one GlcNAc2 Man5 N-glycan chain attached to Asn66. 28489325 0 54 theme Crystal 64:70 arg1 Structure 72:80 the Crystal Structure 60:80 the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin 60:135 Reducing Macro- and Microheterogeneity of N-Glycans Enables the Crystal Structure of the Lectin and EGF-Like Domains of Human L-Selectin To Be Solved at 1.9 Å Resolution. 28489325 5 55 theme " 1052:1052 arg1 conformation 1054:1065 the described "bent" conformation 1033:1065 the described "bent" conformation of E- and P-selectins 1033:1087 The orientation of the lectin and EGF-like domain was similar to the described "bent" conformation of E- and P-selectins. 28489325 1 56 theme most 226:229 arg1 leukocytes 231:240 most leukocytes 226:240 most leukocytes 226:240 L-Selectin, a cell-adhesion receptor on the surface of most leukocytes, contains seven N-glycosylation sites. 25499076 8 0 gly fucosylated 1507:1517 arg1 glycans 1527:1533 fucosylated (90.5%) glycans 1507:1533 fucosylated (90.5%) glycans 1507:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 0 gly fucosylated 1507:1517 arg1 % 1524:1524 90.5% 1520:1524 90.5% 1520:1524 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 2 1 from Asn-31 381:386 arg1 located 370:376 located 370:376 located 370:376 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 6 2 theme ovary 1026:1030 arg1 cells 1032:1036 genetically modified Chinese hamster ovary cells 989:1036 genetically modified Chinese hamster ovary cells (CHO) 989:1042 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 2 theme ovary 1026:1030 arg1 CHO 1039:1041 CHO 1039:1041 CHO 1039:1041 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 5 3 from G-hPRL 888:893 arg1 present 859:865 present 859:865 present 859:865 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 6 4 dep % 1177:1177 arg1 to 1170:1171 to 1170:1171 to 1170:1171 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 5 5 from present 923:929 arg1 hormone 950:956 the recombinant hormone 934:956 the recombinant hormone 934:956 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 6 6 gly glycosylation 1131:1143 arg2 occupancy 1150:1158 the glycosylation site occupancy 1127:1158 the glycosylation site occupancy 1127:1158 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 6 gly glycosylation 1131:1143 arg2 site 1145:1148 the glycosylation site occupancy 1127:1158 the glycosylation site occupancy 1127:1158 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 7 theme % 1168:1168 arg1 % 1177:1177 5.5% to 38.3% 1165:1177 5.5% to 38.3% 1165:1177 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 1 8 theme polypeptide 163:173 arg1 hormone 175:181 a polypeptide hormone 161:181 a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively 161:310 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 8 theme polypeptide 163:173 arg1 prolactin 141:149 Human prolactin 135:149 Human prolactin (hPRL) 135:156 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 5 9 from hormone 950:956 arg1 present 923:929 present 923:929 present 923:929 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 10 10 from content 1754:1760 arg1 agreement 1817:1825 good agreement 1812:1825 good agreement with the values obtained directly via MALDI-TOF-MS 1812:1876 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 9 11 theme in 1552:1553 arg1 bioactivity 1561:1571 A ∼6-fold lower in vitro bioactivity 1536:1571 A ∼6-fold lower in vitro bioactivity 1536:1571 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 1 12 dep glycosylated 231:242 arg1 G-hPRL 245:250 G-hPRL 245:250 G-hPRL 245:250 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 10 13 from methodology 1713:1723 arg1 agreement 1817:1825 good agreement 1812:1825 good agreement with the values obtained directly via MALDI-TOF-MS 1812:1876 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 8 14 theme main 1359:1362 arg1 differences 1364:1374 the main differences 1355:1374 the main differences found in the pituitary sample 1355:1404 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 6 15 theme glycosylation 1131:1143 arg1 occupancy 1150:1158 the glycosylation site occupancy 1127:1158 the glycosylation site occupancy 1127:1158 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 5 16 theme N-glycan 839:846 arg1 structures 848:857 the N-glycan structures 835:857 the N-glycan structures present in native, pituitary G-hPRL 835:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 10 17 with agreement 1817:1825 arg1 values 1836:1841 the values 1832:1841 the values obtained directly via MALDI-TOF-MS 1832:1876 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 5 18 theme present 859:865 arg1 structures 848:857 the N-glycan structures 835:857 the N-glycan structures present in native, pituitary G-hPRL 835:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 9 19 theme clearance 1586:1594 arg1 rate 1596:1599 a higher clearance rate 1577:1599 a higher clearance rate in mice 1577:1607 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 10 20 theme N-Glycan 1659:1666 arg1 profiling 1668:1676 N-Glycan profiling 1659:1676 N-Glycan profiling 1659:1676 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 10 20 theme N-Glycan 1659:1666 arg1 methodology 1713:1723 a useful and accurate methodology 1691:1723 a useful and accurate methodology also for MM 1691:1735 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 8 21 located found 1376:1380 arg1 sample 1399:1404 the pituitary sample 1385:1404 the pituitary sample 1385:1404 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 21 located found 1376:1380 arg2 differences 1364:1374 the main differences 1355:1374 the main differences found in the pituitary sample 1355:1404 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 2 22 contain has 316:318 arg1 It 313:314 It 313:314 It 313:314 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 2 22 contain has 316:318 arg2 site 365:368 a single, partially occupied N-glycosylation site 320:368 a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies 320:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 1 23 gly glycosylated 231:242 arg1 forms 253:257 glycosylated (G-hPRL) forms 231:257 glycosylated (G-hPRL) forms 231:257 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 5 24 attach present 923:929 arg1 hormone 950:956 the recombinant hormone 934:956 the recombinant hormone 934:956 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 5 24 attach present 923:929 arg2 those 917:921 those 917:921 those 917:921 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 8 25 theme low 1424:1426 arg1 presence 1428:1435 an extremely low presence 1411:1435 an extremely low presence of sialylated (1.7%) 1411:1456 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 2 26 gly N-glycosylation 349:363 arg2 site 365:368 a single, partially occupied N-glycosylation site 320:368 a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies 320:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 0 27 theme glycosylated 105:116 arg1 prolactin 124:132 recombinant and pituitary glycosylated human prolactin 79:132 recombinant and pituitary glycosylated human prolactin 79:132 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 6 28 theme modified 1001:1008 arg1 cells 1032:1036 genetically modified Chinese hamster ovary cells 989:1036 genetically modified Chinese hamster ovary cells (CHO) 989:1042 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 28 theme modified 1001:1008 arg1 CHO 1039:1041 CHO 1039:1041 CHO 1039:1041 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 0 29 theme N-glycoprofiling 0:15 arg1 analysis 17:24 N-glycoprofiling analysis 0:24 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 0:133 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 5 30 theme native 870:875 arg1 G-hPRL 888:893 native, pituitary G-hPRL 870:893 native, pituitary G-hPRL 870:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 7 31 theme cell-derived 1226:1237 arg1 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 7 31 theme cell-derived 1226:1237 arg1 G-hPRL 1239:1244 CHO cell-derived G-hPRL 1222:1244 CHO cell-derived G-hPRL (CHO-G-hPRL) 1222:1257 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 6 32 theme recombinant 969:979 arg1 G-hPRL 981:986 recombinant G-hPRL 969:986 recombinant G-hPRL 969:986 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 2 33 theme interesting 430:440 arg1 model 442:446 a particularly simple and interesting model 404:446 a particularly simple and interesting model for glycosylation studies 404:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 10 34 theme useful 1693:1698 arg1 methodology 1713:1723 a useful and accurate methodology 1691:1723 a useful and accurate methodology also for MM 1691:1735 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 10 34 theme useful 1693:1698 arg1 profiling 1668:1676 N-Glycan profiling 1659:1676 N-Glycan profiling 1659:1676 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 0 35 theme simple 31:36 arg1 model 51:55 a simple glycoprotein model 29:55 a simple glycoprotein model 29:55 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 5 36 theme recombinant 938:948 arg1 hormone 950:956 the recombinant hormone 934:956 the recombinant hormone 934:956 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 2 37 theme simple 419:424 arg1 model 442:446 a particularly simple and interesting model 404:446 a particularly simple and interesting model for glycosylation studies 404:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 7 38 link cell-derived 1226:1237 arg1 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 7 38 link cell-derived 1226:1237 arg1 G-hPRL 1239:1244 CHO cell-derived G-hPRL 1222:1244 CHO cell-derived G-hPRL (CHO-G-hPRL) 1222:1257 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 8 39 theme high 1464:1467 arg1 percentage 1469:1478 a high percentage 1462:1478 a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans 1462:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 4 40 theme glycohormones 730:742 arg1 secretion 675:683 secretion 675:683 secretion 675:683 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 4 40 theme glycohormones 730:742 arg1 activity 697:704 biological activity 686:704 biological activity 686:704 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 4 40 theme glycohormones 730:742 arg1 biosynthesis 661:672 biosynthesis 661:672 biosynthesis 661:672 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 4 40 theme glycohormones 730:742 arg1 survival 718:725 plasma survival 711:725 plasma survival 711:725 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 8 41 theme fucosylated 1507:1517 arg1 glycans 1527:1533 fucosylated (90.5%) glycans 1507:1533 fucosylated (90.5%) glycans 1507:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 41 theme fucosylated 1507:1517 arg1 % 1524:1524 90.5% 1520:1524 90.5% 1520:1524 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 0 42 gly glycoprotein 38:49 arg1 glycoprotein 38:49 a simple glycoprotein model 29:55 a simple glycoprotein model 29:55 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 10 43 theme accurate 1704:1711 arg1 methodology 1713:1723 a useful and accurate methodology 1691:1723 a useful and accurate methodology also for MM 1691:1735 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 10 43 theme accurate 1704:1711 arg1 profiling 1668:1676 N-Glycan profiling 1659:1676 N-Glycan profiling 1659:1676 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 10 44 theme carbohydrate 1741:1752 arg1 content 1754:1760 carbohydrate content determination 1741:1774 carbohydrate content determination for the two G-hPRL preparations 1741:1806 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 5 45 attach present 859:865 arg2 structures 848:857 the N-glycan structures 835:857 the N-glycan structures present in native, pituitary G-hPRL 835:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 5 45 attach present 859:865 arg1 G-hPRL 888:893 native, pituitary G-hPRL 870:893 native, pituitary G-hPRL 870:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 6 46 theme hamster 1018:1024 arg1 cells 1032:1036 genetically modified Chinese hamster ovary cells 989:1036 genetically modified Chinese hamster ovary cells (CHO) 989:1042 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 46 theme hamster 1018:1024 arg1 CHO 1039:1041 CHO 1039:1041 CHO 1039:1041 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 3 47 theme G-hPRL 494:499 arg1 lower 504:508 lower 504:508 lower 504:508 The bioactivity of G-hPRL is lower than that of NG-hPRL (by ca. 4-fold) and its physiological function is not clear. 25499076 3 47 theme G-hPRL 494:499 arg1 bioactivity 479:489 The bioactivity 475:489 The bioactivity of G-hPRL 475:499 The bioactivity of G-hPRL is lower than that of NG-hPRL (by ca. 4-fold) and its physiological function is not clear. 25499076 5 48 theme main 789:792 arg1 objective 794:802 The main objective 785:802 The main objective of this study 785:816 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 4 49 theme host 774:777 arg1 cell 779:782 the host cell 770:782 the host cell 770:782 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 3 50 dep 4-fold 539:544 arg1 ca. 535:537 ca. 4-fold 535:544 ca. 4-fold 535:544 The bioactivity of G-hPRL is lower than that of NG-hPRL (by ca. 4-fold) and its physiological function is not clear. 25499076 4 51 theme carbohydrate 601:612 arg1 moieties 614:621 carbohydrate moieties 601:621 carbohydrate moieties 601:621 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 9 52 theme lower 1546:1550 arg1 bioactivity 1561:1571 A ∼6-fold lower in vitro bioactivity 1536:1571 A ∼6-fold lower in vitro bioactivity 1536:1571 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 5 53 dep native 870:875 arg1 pituitary 878:886 pituitary 878:886 pituitary 878:886 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 10 54 theme G-hPRL 1788:1793 arg1 preparations 1795:1806 the two G-hPRL preparations 1780:1806 the two G-hPRL preparations 1780:1806 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 6 55 theme site 1145:1148 arg1 occupancy 1150:1158 the glycosylation site occupancy 1127:1158 the glycosylation site occupancy 1127:1158 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 7 56 theme pituitary 1275:1283 arg1 pit-G-hPRL 1293:1302 pit-G-hPRL 1293:1302 pit-G-hPRL 1293:1302 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 7 56 theme pituitary 1275:1283 arg1 G-hPRL 1285:1290 pituitary G-hPRL 1275:1290 pituitary G-hPRL (pit-G-hPRL) 1275:1303 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 10 57 theme good 1812:1815 arg1 agreement 1817:1825 good agreement 1812:1825 good agreement with the values obtained directly via MALDI-TOF-MS 1812:1876 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 6 58 theme G-hPRL 1201:1206 arg1 purification 1208:1219 G-hPRL purification 1201:1219 G-hPRL purification 1201:1219 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 3 59 theme physiological 555:567 arg1 function 569:576 its physiological function 551:576 its physiological function 551:576 The bioactivity of G-hPRL is lower than that of NG-hPRL (by ca. 4-fold) and its physiological function is not clear. 25499076 1 60 theme Human 135:139 arg1 hormone 175:181 a polypeptide hormone 161:181 a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively 161:310 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 60 theme Human 135:139 arg1 hPRL 152:155 hPRL 152:155 hPRL 152:155 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 60 theme Human 135:139 arg1 prolactin 141:149 Human prolactin 135:149 Human prolactin (hPRL) 135:156 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 9 61 theme higher 1579:1584 arg1 rate 1596:1599 a higher clearance rate 1577:1599 a higher clearance rate in mice 1577:1607 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 8 62 theme pituitary 1389:1397 arg1 sample 1399:1404 the pituitary sample 1385:1404 the pituitary sample 1385:1404 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 9 63 dep in 1552:1553 arg1 vitro 1555:1559 vitro 1555:1559 vitro 1555:1559 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 5 64 theme study 812:816 arg1 objective 794:802 The main objective 785:802 The main objective of this study 785:816 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 9 65 from bioactivity 1561:1571 arg1 mice 1604:1607 mice 1604:1607 mice 1604:1607 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 2 66 theme glycosylation 452:464 arg1 studies 466:472 glycosylation studies 452:472 glycosylation studies 452:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 0 67 theme recombinant 79:89 arg1 prolactin 124:132 recombinant and pituitary glycosylated human prolactin 79:132 recombinant and pituitary glycosylated human prolactin 79:132 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 0 68 from analysis 17:24 arg1 model 51:55 a simple glycoprotein model 29:55 a simple glycoprotein model 29:55 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 4 69 theme important 638:646 arg1 roles 648:652 important roles 638:652 important roles 638:652 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 0 70 theme pituitary 95:103 arg1 prolactin 124:132 recombinant and pituitary glycosylated human prolactin 79:132 recombinant and pituitary glycosylated human prolactin 79:132 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 0 71 theme human 118:122 arg1 prolactin 124:132 recombinant and pituitary glycosylated human prolactin 79:132 recombinant and pituitary glycosylated human prolactin 79:132 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 6 72 theme Chinese 1010:1016 arg1 cells 1032:1036 genetically modified Chinese hamster ovary cells 989:1036 genetically modified Chinese hamster ovary cells (CHO) 989:1042 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 6 72 theme Chinese 1010:1016 arg1 CHO 1039:1041 CHO 1039:1041 CHO 1039:1041 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 8 73 theme sialylated 1440:1449 arg1 presence 1428:1435 an extremely low presence 1411:1435 an extremely low presence of sialylated (1.7%) 1411:1456 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 73 theme sialylated 1440:1449 arg1 percentage 1469:1478 a high percentage 1462:1478 a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans 1462:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 2 74 dep single 322:327 arg1 occupied 340:347 occupied 340:347 occupied 340:347 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 2 75 theme N-glycosylation 349:363 arg1 site 365:368 a single, partially occupied N-glycosylation site 320:368 a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies 320:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 5 76 from present 859:865 arg1 G-hPRL 888:893 native, pituitary G-hPRL 870:893 native, pituitary G-hPRL 870:893 The main objective of this study was to determine the N-glycan structures present in native, pituitary G-hPRL and compare them with those present in the recombinant hormone. 25499076 7 77 theme CHO 1222:1224 arg1 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO-G-hPRL 1247:1256 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 7 77 theme CHO 1222:1224 arg1 G-hPRL 1239:1244 CHO cell-derived G-hPRL 1222:1244 CHO cell-derived G-hPRL (CHO-G-hPRL) 1222:1257 CHO cell-derived G-hPRL (CHO-G-hPRL) was compared to pituitary G-hPRL (pit-G-hPRL) especially with regard to N-glycoprofiling. 25499076 6 78 from growth 1056:1061 arg1 suspension 1066:1075 suspension 1066:1075 suspension 1066:1075 To obtain recombinant G-hPRL, genetically modified Chinese hamster ovary cells (CHO), adapted to growth in suspension, were treated with cycloheximide, thus increasing the glycosylation site occupancy from 5.5% to 38.3%, thereby facilitating G-hPRL purification. 25499076 0 79 gly glycosylated 105:116 arg1 prolactin 124:132 recombinant and pituitary glycosylated human prolactin 79:132 recombinant and pituitary glycosylated human prolactin 79:132 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 4 80 theme biological 686:695 arg1 activity 697:704 biological activity 686:704 biological activity 686:704 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 0 81 theme glycoprotein 38:49 arg1 model 51:55 a simple glycoprotein model 29:55 a simple glycoprotein model 29:55 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 2 82 theme located 370:376 arg1 site 365:368 a single, partially occupied N-glycosylation site 320:368 a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies 320:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 1 83 theme 23 285:286 arg1 MM 265:266 MM 265:266 MM of approximately 23 265:286 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 83 theme 23 285:286 arg1 25kDa 292:296 25kDa 292:296 25kDa 292:296 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 84 with forms 253:257 arg1 MM 265:266 MM 265:266 MM of approximately 23 265:286 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 1 84 with forms 253:257 arg1 25kDa 292:296 25kDa 292:296 25kDa 292:296 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 25499076 10 85 dep content 1754:1760 arg1 determination 1762:1774 determination 1762:1774 carbohydrate content determination for the two G-hPRL preparations 1741:1806 N-Glycan profiling proved to be a useful and accurate methodology also for MM and carbohydrate content determination for the two G-hPRL preparations, in good agreement with the values obtained directly via MALDI-TOF-MS. 25499076 8 86 theme sulfated 1483:1490 arg1 presence 1428:1435 an extremely low presence 1411:1435 an extremely low presence of sialylated (1.7%) 1411:1456 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 86 theme sulfated 1483:1490 arg1 percentage 1469:1478 a high percentage 1462:1478 a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans 1462:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 4 87 theme plasma 711:716 arg1 survival 718:725 plasma survival 711:725 plasma survival 711:725 However, carbohydrate moieties generally play important roles in the biosynthesis, secretion, biological activity, and plasma survival of glycohormones and can vary depending on the host cell. 25499076 9 88 from rate 1596:1599 arg1 mice 1604:1607 mice 1604:1607 mice 1604:1607 A ∼6-fold lower in vitro bioactivity and a higher clearance rate in mice were also found for pit-G-hPRL versus CHO-G-hPRL. 25499076 0 89 dep analysis 17:24 arg1 comparison 60:69 a comparison 58:69 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 0:133 N-glycoprofiling analysis in a simple glycoprotein model: a comparison between recombinant and pituitary glycosylated human prolactin. 25499076 2 90 theme single 322:327 arg1 site 365:368 a single, partially occupied N-glycosylation site 320:368 a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies 320:472 It has a single, partially occupied N-glycosylation site located at Asn-31, which makes it a particularly simple and interesting model for glycosylation studies. 25499076 8 91 theme glycans 1527:1533 arg1 presence 1428:1435 an extremely low presence 1411:1435 an extremely low presence of sialylated (1.7%) 1411:1456 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 8 91 theme glycans 1527:1533 arg1 percentage 1469:1478 a high percentage 1462:1478 a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans 1462:1533 Among the main differences found in the pituitary sample were an extremely low presence of sialylated (1.7%) and a high percentage of sulfated (74.0%) and of fucosylated (90.5%) glycans. 25499076 1 92 theme glycosylated 231:242 arg1 forms 253:257 glycosylated (G-hPRL) forms 231:257 glycosylated (G-hPRL) forms 231:257 Human prolactin (hPRL) is a polypeptide hormone occurring in the non-glycosylated (NG-hPRL) and glycosylated (G-hPRL) forms, with MM of approximately 23 and 25kDa, respectively. 26339047 5 0 contain carry 1031:1035 arg2 types 1046:1050 the same types 1037:1050 the same types of glycans as WT VLPs 1037:1072 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 5 0 contain carry 1031:1035 arg1 VLPs 1026:1029 the hyperglycosylated VLPs 1004:1029 the hyperglycosylated VLPs 1004:1029 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 3 1 theme potential 698:706 arg1 sites 722:726 potential glycosylation sites 698:726 potential glycosylation sites 698:726 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 13 2 theme wild-type 2685:2693 arg1 VLPs 2715:2718 wild-type or hypoglycosylated VLPs 2685:2718 wild-type or hypoglycosylated VLPs 2685:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 11 3 gly N-glycosylation 2351:2365 arg2 sites 2367:2371 additional N-glycosylation sites 2340:2371 additional N-glycosylation sites 2340:2371 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 6 4 theme WT 1215:1216 arg1 VLPs 1250:1253 the WT and hypo- and hyperglycosylated VLPs 1211:1253 the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies 1211:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 8 5 theme hypoglycosylated 1715:1730 arg1 VLPs 1739:1742 the hypoglycosylated and WT VLPs 1711:1742 the hypoglycosylated and WT VLPs 1711:1742 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 6 6 theme antibodies 1298:1307 arg1 panel 1263:1267 a panel 1261:1267 a panel of 19 anti-HBsAgS monoclonal antibodies 1261:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 3 7 theme VLPs 577:580 arg1 generation 546:555 the generation 542:555 the generation of hypoglycosylated VLPs 542:580 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 1 8 theme HBV 252:254 arg1 vaccine 256:262 the HBV vaccine 248:262 the HBV vaccine 248:262 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 4 9 theme T116N 849:853 arg1 anchors 902:908 glycosylation anchors 888:908 glycosylation anchors resulting in the formation of hyperglycosylated VLPs 888:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 9 theme T116N 849:853 arg1 sites 865:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 9 10 theme preventive 1976:1985 arg1 applications 1987:1998 superior preventive applications 1967:1998 superior preventive applications 1967:1998 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 6 11 theme anti-HBsAgS 1275:1285 arg1 antibodies 1298:1307 19 anti-HBsAgS monoclonal antibodies 1272:1307 19 anti-HBsAgS monoclonal antibodies 1272:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 4 12 theme G130N 859:863 arg1 anchors 902:908 glycosylation anchors 888:908 glycosylation anchors resulting in the formation of hyperglycosylated VLPs 888:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 12 theme G130N 859:863 arg1 sites 865:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 13 used utilized 876:883 arg2 sites 865:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 13 used utilized 876:883 arg2 anchors 902:908 glycosylation anchors 888:908 glycosylation anchors resulting in the formation of hyperglycosylated VLPs 888:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 6 14 theme Antigenic 1184:1192 arg1 fingerprints 1194:1205 Antigenic fingerprints 1184:1205 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies 1184:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 0 15 with Particles 96:104 arg1 Immunogenicity 120:133 Enhanced Immunogenicity 111:133 Enhanced Immunogenicity 111:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 1 16 theme hepatitis 186:194 arg1 virus 198:202 hepatitis B virus 186:202 hepatitis B virus (HBV) 186:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 16 theme hepatitis 186:194 arg1 HBV 205:207 HBV 205:207 HBV 205:207 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 9 17 theme VLPs 1770:1773 arg1 ability 1749:1755 The ability 1745:1755 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system 1745:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 13 18 theme immunization 2535:2546 arg1 studies 2548:2554 Comparative immunization studies 2523:2554 Comparative immunization studies 2523:2554 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 3 19 theme external 758:765 arg1 region 772:777 the HBsAgS external loop region 747:777 the HBsAgS external loop region 747:777 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 13 20 theme earlier 2624:2630 arg1 responses 2667:2675 earlier and longer-lasting antibody immune responses 2624:2675 earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs 2624:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 13 21 with VLPs 2578:2581 arg1 density 2609:2615 the highest N-glycan density 2588:2615 the highest N-glycan density 2588:2615 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 4 22 theme hyperglycosylated 940:956 arg1 VLPs 958:961 hyperglycosylated VLPs 940:961 hyperglycosylated VLPs 940:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 13 23 theme longer-lasting 2636:2649 arg1 responses 2667:2675 earlier and longer-lasting antibody immune responses 2624:2675 earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs 2624:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 13 24 dep do 2682:2683 arg1 than 2677:2680 than 2677:2680 than 2677:2680 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 13 25 theme highest 2592:2598 arg1 density 2609:2615 the highest N-glycan density 2588:2615 the highest N-glycan density 2588:2615 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 6 26 theme hyperglycosylated 1232:1248 arg1 VLPs 1250:1253 the WT and hypo- and hyperglycosylated VLPs 1211:1253 the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies 1211:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 4 27 gly hyperglycosylated 940:956 arg1 VLPs 958:961 hyperglycosylated VLPs 940:961 hyperglycosylated VLPs 940:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 2 28 theme glycoengineered 514:528 arg1 HBsAgS 530:535 glycoengineered HBsAgS 514:535 glycoengineered HBsAgS 514:535 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 6 29 theme hypo- 1222:1226 arg1 VLPs 1250:1253 the WT and hypo- and hyperglycosylated VLPs 1211:1253 the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies 1211:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 12 30 theme glycans 2496:2502 arg1 types 2487:2491 the same types 2478:2491 the same types of glycans as wild-type VLPs 2478:2520 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 1 31 theme foreign 302:308 arg1 sequences 320:328 foreign antigenic sequences 302:328 foreign antigenic sequences 302:328 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 13 32 theme vaccine 2758:2764 arg1 injections 2766:2775 vaccine injections 2758:2775 vaccine injections 2758:2775 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 12 33 gly Hyperglycosylated 2374:2390 arg1 VLPs 2392:2395 Hyperglycosylated VLPs 2374:2395 Hyperglycosylated VLPs 2374:2395 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 0 34 theme B 77:77 arg1 Particles 96:104 Hepatitis B Virus Virus-Like Particles 67:104 Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity 67:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 1 35 theme sequences 320:328 arg1 delivery 290:297 the delivery 286:297 the delivery of foreign antigenic sequences 286:328 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 13 36 theme hypoglycosylated 2698:2713 arg1 VLPs 2715:2718 wild-type or hypoglycosylated VLPs 2685:2718 wild-type or hypoglycosylated VLPs 2685:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 11 37 theme N-glycosylation 2351:2365 arg1 sites 2367:2371 additional N-glycosylation sites 2340:2371 additional N-glycosylation sites 2340:2371 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 3 38 theme WT 825:826 arg1 site 828:831 the WT site 821:831 the WT site 821:831 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 0 39 theme Virus-Like 85:94 arg1 Particles 96:104 Hepatitis B Virus Virus-Like Particles 67:104 Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity 67:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 3 40 dep positions 782:790 arg1 T116 792:795 T116 792:795 T116 792:795 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 3 40 dep positions 782:790 arg1 positions 782:790 positions T116 and G130 782:804 positions T116 and G130 in addition to the WT site 782:831 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 3 40 dep positions 782:790 arg1 G130 801:804 G130 801:804 G130 801:804 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 1 41 theme small 140:144 arg1 proteins 155:162 The small envelope proteins 136:162 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) 136:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 41 theme small 140:144 arg1 HBsAgS 165:170 HBsAgS 165:170 HBsAgS 165:170 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 6 42 theme different 1375:1383 arg1 glyco-analogues 1389:1403 the different VLP glyco-analogues 1371:1403 the different VLP glyco-analogues 1371:1403 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 13 43 dep responses 2667:2675 arg1 do 2682:2683 do 2682:2683 do 2682:2683 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 0 44 theme Hepatitis 67:75 arg1 Particles 96:104 Hepatitis B Virus Virus-Like Particles 67:104 Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity 67:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 10 45 theme modified 2038:2045 arg1 immunogens 2058:2067 biochemically modified, nonnative immunogens 2024:2067 biochemically modified, nonnative immunogens 2024:2067 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 1 46 theme vaccine 256:262 arg1 components 234:243 the antigenic components 220:243 the antigenic components of the HBV vaccine 220:262 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 13 47 theme antibody 2651:2658 arg1 responses 2667:2675 earlier and longer-lasting antibody immune responses 2624:2675 earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs 2624:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 14 48 theme delivery 2902:2909 arg1 platforms 2911:2919 VLP delivery platforms 2898:2919 VLP delivery platforms 2898:2919 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 9 49 theme immune 1899:1904 arg1 system 1906:1911 the innate immune system 1888:1911 the innate immune system 1888:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 0 50 theme Asparagine-Linked 16:32 arg1 Density 41:47 Asparagine-Linked Glycan Density 16:47 Asparagine-Linked Glycan Density 16:47 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 5 51 theme spectroscopic 969:981 arg1 analyses 983:990 Mass spectroscopic analyses 964:990 Mass spectroscopic analyses 964:990 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 11 52 theme virus 2302:2306 arg1 HBsAgS 2327:2332 HBsAgS 2327:2332 HBsAgS 2327:2332 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 11 52 theme virus 2302:2306 arg1 proteins 2317:2324 hepatitis B virus envelope proteins 2290:2324 hepatitis B virus envelope proteins (HBsAgS) 2290:2333 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 10 53 dep modified 2038:2045 arg1 nonnative 2048:2056 nonnative 2048:2056 nonnative 2048:2056 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 2 54 theme immunization 412:423 arg1 vectors 425:431 improved immunization vectors 403:431 improved immunization vectors 403:431 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 0 55 theme Density 41:47 arg1 Modification 0:11 Modification 0:11 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 0:134 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 11 56 theme envelope 2308:2315 arg1 HBsAgS 2327:2332 HBsAgS 2327:2332 HBsAgS 2327:2332 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 11 56 theme envelope 2308:2315 arg1 proteins 2317:2324 hepatitis B virus envelope proteins 2290:2324 hepatitis B virus envelope proteins (HBsAgS) 2290:2333 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 14 57 theme optimized 2875:2883 arg1 vaccines 2885:2892 optimized vaccines 2875:2892 optimized vaccines 2875:2892 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 2 58 theme virus-like 475:484 arg1 VLPs 497:500 VLPs 497:500 VLPs 497:500 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 2 58 theme virus-like 475:484 arg1 particles 486:494 biochemically modified virus-like particles 452:494 biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS 452:535 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 8 59 theme T116N 1637:1641 arg1 VLPs 1643:1646 The T116N VLPs 1633:1646 The T116N VLPs 1633:1646 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 11 60 theme virus-like 2250:2259 arg1 VLPs 2272:2275 VLPs 2272:2275 VLPs 2272:2275 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 11 60 theme virus-like 2250:2259 arg1 particles 2261:2269 virus-like particles 2250:2269 virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) 2250:2333 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 7 61 theme strong 1566:1571 arg1 correlation 1573:1583 a strong correlation 1564:1583 a strong correlation between N-glycan abundance and antibody titers 1564:1630 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 5 62 theme glycans 1055:1061 arg1 types 1046:1050 the same types 1037:1050 the same types of glycans as WT VLPs 1037:1072 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 9 63 from differences 1819:1829 arg1 interaction 1862:1872 their glycosylation-related interaction 1834:1872 their glycosylation-related interaction with cells of the innate immune system 1834:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 9 64 theme immunogens 1952:1961 arg1 design 1942:1947 the design 1938:1947 the design of immunogens 1938:1961 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 3 65 theme hyperglycosylated 674:690 arg1 VLPs 692:695 hyperglycosylated VLPs 674:695 hyperglycosylated VLPs 674:695 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 7 66 with studies 1525:1531 arg1 VLPs 1552:1555 the different VLPs 1538:1555 the different VLPs 1538:1555 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 11 67 theme hepatitis 2290:2298 arg1 virus 2302:2306 hepatitis B virus 2290:2306 hepatitis B virus envelope proteins (HBsAgS) 2290:2333 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 7 68 theme different 1542:1550 arg1 VLPs 1552:1555 the different VLPs 1538:1555 the different VLPs 1538:1555 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 8 69 gly hypoglycosylated 1715:1730 arg1 VLPs 1739:1742 the hypoglycosylated and WT VLPs 1711:1742 the hypoglycosylated and WT VLPs 1711:1742 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 14 70 theme immunogen 2827:2835 arg1 immunogenicity 2806:2819 the immunogenicity 2802:2819 the immunogenicity of an immunogen 2802:2835 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 9 71 theme superior 1967:1974 arg1 applications 1987:1998 superior preventive applications 1967:1998 superior preventive applications 1967:1998 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 8 72 theme antibody 1683:1690 arg1 responses 1692:1700 earlier and longer-lasting antibody responses 1656:1700 earlier and longer-lasting antibody responses 1656:1700 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 5 73 theme bisecting 1135:1143 arg1 N-acetylglucosamines 1145:1164 bisecting N-acetylglucosamines 1135:1164 bisecting N-acetylglucosamines 1135:1164 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 8 74 dep did 1707:1709 arg1 than 1702:1705 than 1702:1705 than 1702:1705 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 12 75 theme same 2482:2485 arg1 types 2487:2491 the same types 2478:2491 the same types of glycans as wild-type VLPs 2478:2520 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 3 76 theme glycosylation 708:720 arg1 sites 722:726 potential glycosylation sites 698:726 potential glycosylation sites 698:726 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 3 77 theme hypoglycosylated 560:575 arg1 VLPs 577:580 hypoglycosylated VLPs 560:580 hypoglycosylated VLPs 560:580 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 14 78 theme adjuvanting 2999:3009 arg1 compounds 3011:3019 suitable adjuvanting compounds 2990:3019 suitable adjuvanting compounds 2990:3019 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 7 79 theme Immunization 1512:1523 arg1 studies 1525:1531 Immunization studies 1512:1531 Immunization studies with the different VLPs 1512:1555 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 10 80 theme immunogens 2058:2067 arg1 use 2017:2019 The use 2013:2019 The use of biochemically modified, nonnative immunogens 2013:2067 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 3 81 theme HBsAgS 751:756 arg1 region 772:777 the HBsAgS external loop region 747:777 the HBsAgS external loop region 747:777 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 6 82 theme monoclonal 1287:1296 arg1 antibodies 1298:1307 19 anti-HBsAgS monoclonal antibodies 1272:1307 19 anti-HBsAgS monoclonal antibodies 1272:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 12 83 theme wild-type 2507:2515 arg1 VLPs 2517:2520 wild-type VLPs 2507:2520 wild-type VLPs 2507:2520 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 11 84 theme B 2300:2300 arg1 virus 2302:2306 hepatitis B virus 2290:2306 hepatitis B virus envelope proteins (HBsAgS) 2290:2333 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 4 85 theme introduced 838:847 arg1 anchors 902:908 glycosylation anchors 888:908 glycosylation anchors resulting in the formation of hyperglycosylated VLPs 888:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 85 theme introduced 838:847 arg1 sites 865:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 14 86 theme compounds 3011:3019 arg1 use 2983:2985 the use 2979:2985 the use of suitable adjuvanting compounds 2979:3019 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 8 87 theme earlier 1656:1662 arg1 responses 1692:1700 earlier and longer-lasting antibody responses 1656:1700 earlier and longer-lasting antibody responses 1656:1700 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 3 88 theme wild-type 587:595 arg1 site 628:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 5 89 theme N-acetylglucosamines 1145:1164 arg1 degree 1111:1116 the degree 1107:1116 the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation 1107:1181 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 9 90 theme nonnative 1760:1768 arg1 VLPs 1770:1773 nonnative VLPs 1760:1773 nonnative VLPs 1760:1773 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 14 91 theme suitable 2990:2997 arg1 compounds 3011:3019 suitable adjuvanting compounds 2990:3019 suitable adjuvanting compounds 2990:3019 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 3 92 gly glycosylation 614:626 arg2 site 628:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 3 92 gly glycosylation 614:626 arg2 N146 609:612 N146 609:612 N146 609:612 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 13 93 theme immune 2660:2665 arg1 responses 2667:2675 earlier and longer-lasting antibody immune responses 2624:2675 earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs 2624:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 1 94 theme B 196:196 arg1 virus 198:202 hepatitis B virus 186:202 hepatitis B virus (HBV) 186:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 94 theme B 196:196 arg1 HBV 205:207 HBV 205:207 HBV 205:207 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 12 95 contain carry 2472:2476 arg1 they 2467:2470 they 2467:2470 they 2467:2470 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 12 95 contain carry 2472:2476 arg2 types 2487:2491 the same types 2478:2491 the same types of glycans as wild-type VLPs 2478:2520 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 3 96 theme loop 767:770 arg1 region 772:777 the HBsAgS external loop region 747:777 the HBsAgS external loop region 747:777 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 5 97 theme minor 1080:1084 arg1 variations 1086:1095 minor variations 1080:1095 minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation 1080:1181 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 3 98 theme HBsAgS 602:607 arg1 site 628:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 9 99 theme immune 1786:1791 arg1 responses 1793:1801 immune responses 1786:1801 immune responses 1786:1801 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 13 100 theme injections 2766:2775 arg1 numbers 2747:2753 reduced numbers 2739:2753 reduced numbers of vaccine injections 2739:2775 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 10 101 with interaction 2201:2211 arg1 cells 2225:2229 immune cells 2218:2229 immune cells 2218:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 9 102 with interaction 1862:1872 arg1 cells 1879:1883 cells 1879:1883 cells of the innate immune system 1879:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 4 103 theme glycosylation 888:900 arg1 anchors 902:908 glycosylation anchors 888:908 glycosylation anchors resulting in the formation of hyperglycosylated VLPs 888:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 4 103 theme glycosylation 888:900 arg1 sites 865:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites 834:869 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 10 104 theme attractive 2083:2092 arg1 strategy 2094:2101 an attractive strategy 2080:2101 an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells 2080:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 13 105 theme N-glycan 2600:2607 arg1 density 2609:2615 the highest N-glycan density 2588:2615 the highest N-glycan density 2588:2615 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 5 106 theme WT 1066:1067 arg1 VLPs 1069:1072 WT VLPs 1066:1072 WT VLPs 1066:1072 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 13 107 theme reduced 2739:2745 arg1 numbers 2747:2753 reduced numbers 2739:2753 reduced numbers of vaccine injections 2739:2775 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 10 108 theme modulated 2125:2133 arg1 responses 2154:2162 modulated or enhanced immune responses 2125:2162 modulated or enhanced immune responses possibly due to differences in their interaction with immune cells 2125:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 6 109 theme additional 1426:1435 arg1 N-glycans 1437:1445 the additional N-glycans 1422:1445 the additional N-glycans 1422:1445 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 3 110 theme glycosylation 614:626 arg1 site 628:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 the wild-type (WT) HBsAgS N146 glycosylation site 583:631 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 1 111 theme envelope 146:153 arg1 proteins 155:162 The small envelope proteins 136:162 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) 136:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 111 theme envelope 146:153 arg1 HBsAgS 165:170 HBsAgS 165:170 HBsAgS 165:170 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 10 112 theme immune 2147:2152 arg1 responses 2154:2162 modulated or enhanced immune responses 2125:2162 modulated or enhanced immune responses possibly due to differences in their interaction with immune cells 2125:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 14 113 theme antigenic 2933:2941 arg1 sequences 2943:2951 foreign antigenic sequences 2925:2951 foreign antigenic sequences 2925:2951 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 3 114 gly hypoglycosylated 560:575 arg1 VLPs 577:580 hypoglycosylated VLPs 560:580 hypoglycosylated VLPs 560:580 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 0 115 theme Virus 79:83 arg1 Particles 96:104 Hepatitis B Virus Virus-Like Particles 67:104 Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity 67:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 1 116 theme antigenic 310:318 arg1 sequences 320:328 foreign antigenic sequences 302:328 foreign antigenic sequences 302:328 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 3 117 dep wild-type 587:595 arg1 WT 598:599 WT 598:599 WT 598:599 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 9 118 theme glycosylation-related 1840:1860 arg1 interaction 1862:1872 their glycosylation-related interaction 1834:1872 their glycosylation-related interaction with cells of the innate immune system 1834:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 10 119 theme responses 2154:2162 arg1 generation 2111:2120 the generation 2107:2120 the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells 2107:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 0 120 theme Particles 96:104 arg1 Design 57:62 the Design 53:62 the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity 53:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 7 121 theme antibody 1616:1623 arg1 titers 1625:1630 antibody titers 1616:1630 antibody titers 1616:1630 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 4 122 theme VLPs 958:961 arg1 formation 927:935 the formation 923:935 the formation of hyperglycosylated VLPs 923:961 The introduced T116N and G130N sites were utilized as glycosylation anchors resulting in the formation of hyperglycosylated VLPs. 26339047 12 123 theme Hyperglycosylated 2374:2390 arg1 VLPs 2392:2395 Hyperglycosylated VLPs 2374:2395 Hyperglycosylated VLPs 2374:2395 Hyperglycosylated VLPs were synthesized and characterized, and the results demonstrated that they carry the same types of glycans as wild-type VLPs. 26339047 10 124 theme enhanced 2138:2145 arg1 responses 2154:2162 modulated or enhanced immune responses 2125:2162 modulated or enhanced immune responses possibly due to differences in their interaction with immune cells 2125:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 0 125 theme Enhanced 111:118 arg1 Immunogenicity 120:133 Enhanced Immunogenicity 111:133 Enhanced Immunogenicity 111:133 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 13 126 gly hypoglycosylated 2698:2713 arg1 VLPs 2715:2718 wild-type or hypoglycosylated VLPs 2685:2718 wild-type or hypoglycosylated VLPs 2685:2718 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 14 127 theme foreign 2925:2931 arg1 sequences 2943:2951 foreign antigenic sequences 2925:2951 foreign antigenic sequences 2925:2951 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 11 128 theme additional 2340:2349 arg1 sites 2367:2371 additional N-glycosylation sites 2340:2371 additional N-glycosylation sites 2340:2371 We have generated virus-like particles (VLPs) composed of hepatitis B virus envelope proteins (HBsAgS) with additional N-glycosylation sites. 26339047 10 129 from differences 2180:2190 arg1 interaction 2201:2211 their interaction 2195:2211 their interaction with immune cells 2195:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 6 130 theme VLP 1385:1387 arg1 glyco-analogues 1389:1403 the different VLP glyco-analogues 1371:1403 the different VLP glyco-analogues 1371:1403 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 6 131 theme HBsAgS-specific 1482:1496 arg1 antigenicity 1498:1509 the HBsAgS-specific antigenicity 1478:1509 the HBsAgS-specific antigenicity 1478:1509 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 5 132 theme fucosylation 1121:1132 arg1 degree 1111:1116 the degree 1107:1116 the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation 1107:1181 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 13 133 theme Comparative 2523:2533 arg1 studies 2548:2554 Comparative immunization studies 2523:2554 Comparative immunization studies 2523:2554 Comparative immunization studies demonstrated that the VLPs with the highest N-glycan density induce earlier and longer-lasting antibody immune responses than do wild-type or hypoglycosylated VLPs, possibly allowing reduced numbers of vaccine injections. 26339047 9 134 theme innate 1892:1897 arg1 system 1906:1911 the innate immune system 1888:1911 the innate immune system 1888:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 0 135 theme Glycan 34:39 arg1 Density 41:47 Asparagine-Linked Glycan Density 16:47 Asparagine-Linked Glycan Density 16:47 Modification of Asparagine-Linked Glycan Density for the Design of Hepatitis B Virus Virus-Like Particles with Enhanced Immunogenicity. 26339047 5 136 theme Mass 964:967 arg1 analyses 983:990 Mass spectroscopic analyses 964:990 Mass spectroscopic analyses 964:990 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 14 137 theme VLP 2898:2900 arg1 platforms 2911:2919 VLP delivery platforms 2898:2919 VLP delivery platforms 2898:2919 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 9 138 theme system 1906:1911 arg1 cells 1879:1883 cells 1879:1883 cells of the innate immune system 1879:1911 The ability of nonnative VLPs to promote immune responses possibly due to differences in their glycosylation-related interaction with cells of the innate immune system illustrates pathways for the design of immunogens for superior preventive applications. 26339047 1 139 attach derived 173:179 arg2 HBsAgS 165:170 HBsAgS 165:170 HBsAgS 165:170 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 139 attach derived 173:179 arg1 virus 198:202 hepatitis B virus 186:202 hepatitis B virus (HBV) 186:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 139 attach derived 173:179 arg2 proteins 155:162 The small envelope proteins 136:162 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) 136:208 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 1 139 attach derived 173:179 arg1 HBV 205:207 HBV 205:207 HBV 205:207 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 26339047 2 140 theme vectors 425:431 arg1 design 393:398 the design 389:398 the design of improved immunization vectors 389:431 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 5 141 theme same 1041:1044 arg1 types 1046:1050 the same types 1037:1050 the same types of glycans as WT VLPs 1037:1072 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 5 142 gly hyperglycosylated 1008:1024 arg1 VLPs 1026:1029 the hyperglycosylated VLPs 1004:1029 the hyperglycosylated VLPs 1004:1029 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 2 143 theme improved 403:410 arg1 vectors 425:431 improved immunization vectors 403:431 improved immunization vectors 403:431 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 5 144 theme sialylation 1171:1181 arg1 degree 1111:1116 the degree 1107:1116 the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation 1107:1181 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 7 145 theme N-glycan 1593:1600 arg1 abundance 1602:1610 N-glycan abundance 1593:1610 N-glycan abundance 1593:1610 Immunization studies with the different VLPs showed a strong correlation between N-glycan abundance and antibody titers. 26339047 2 146 theme modified 466:473 arg1 VLPs 497:500 VLPs 497:500 VLPs 497:500 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 2 146 theme modified 466:473 arg1 particles 486:494 biochemically modified virus-like particles 452:494 biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS 452:535 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 3 147 gly glycosylation 708:720 arg2 sites 722:726 potential glycosylation sites 698:726 potential glycosylation sites 698:726 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 14 148 with synergy 2966:2972 arg1 use 2983:2985 the use 2979:2985 the use of suitable adjuvanting compounds 2979:3019 The ability to modulate the immunogenicity of an immunogen will provide opportunities to develop optimized vaccines and VLP delivery platforms for foreign antigenic sequences, possibly in synergy with the use of suitable adjuvanting compounds. 26339047 8 149 theme WT 1736:1737 arg1 VLPs 1739:1742 the hypoglycosylated and WT VLPs 1711:1742 the hypoglycosylated and WT VLPs 1711:1742 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 5 150 theme hyperglycosylated 1008:1024 arg1 VLPs 1026:1029 the hyperglycosylated VLPs 1004:1029 the hyperglycosylated VLPs 1004:1029 Mass spectroscopic analyses showed that the hyperglycosylated VLPs carry the same types of glycans as WT VLPs, with minor variations regarding the degree of fucosylation, bisecting N-acetylglucosamines, and sialylation. 26339047 8 151 theme longer-lasting 1668:1681 arg1 responses 1692:1700 earlier and longer-lasting antibody responses 1656:1700 earlier and longer-lasting antibody responses 1656:1700 The T116N VLPs induced earlier and longer-lasting antibody responses than did the hypoglycosylated and WT VLPs. 26339047 10 152 theme immune 2218:2223 arg1 cells 2225:2229 immune cells 2218:2229 immune cells 2218:2229 IMPORTANCE: The use of biochemically modified, nonnative immunogens represents an attractive strategy for the generation of modulated or enhanced immune responses possibly due to differences in their interaction with immune cells. 26339047 3 153 gly hyperglycosylated 674:690 arg1 VLPs 692:695 hyperglycosylated VLPs 674:695 hyperglycosylated VLPs 674:695 For the generation of hypoglycosylated VLPs, the wild-type (WT) HBsAgS N146 glycosylation site was converted to N146Q; for constructing hyperglycosylated VLPs, potential glycosylation sites were introduced in the HBsAgS external loop region at positions T116 and G130 in addition to the WT site. 26339047 6 154 gly hyperglycosylated 1232:1248 arg1 VLPs 1250:1253 the WT and hypo- and hyperglycosylated VLPs 1211:1253 the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies 1211:1307 Antigenic fingerprints for the WT and hypo- and hyperglycosylated VLPs using a panel of 19 anti-HBsAgS monoclonal antibodies revealed that 15 antibodies retained their ability to bind to the different VLP glyco-analogues, suggesting that the additional N-glycans did not shield extensively for the HBsAgS-specific antigenicity. 26339047 2 155 theme structure-immunogenicity 346:369 arg1 relationships 371:383 structure-immunogenicity relationships 346:383 structure-immunogenicity relationships for the design of improved immunization vectors 346:431 To investigate structure-immunogenicity relationships for the design of improved immunization vectors, we have generated biochemically modified virus-like particles (VLPs) exhibiting glycoengineered HBsAgS. 26339047 1 156 theme antigenic 224:232 arg1 components 234:243 the antigenic components 220:243 the antigenic components of the HBV vaccine 220:262 The small envelope proteins (HBsAgS) derived from hepatitis B virus (HBV) represent the antigenic components of the HBV vaccine and are platforms for the delivery of foreign antigenic sequences. 27884519 11 0 theme wild 1303:1306 arg1 WT 1314:1315 WT 1314:1315 WT 1314:1315 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 11 0 theme wild 1303:1306 arg1 type 1308:1311 wild type 1303:1311 wild type (WT) 1303:1316 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 6 1 theme large 791:795 arg1 system 814:819 A large scale production system 789:819 A large scale production system 789:819 A large scale production system was established and the characteristics of modified hIL-11s were evaluated. 27884519 10 2 gly glycosylated 1185:1196 arg1 proteins 1166:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins were well glycosylated and produced. 27884519 9 3 theme biological 1078:1087 arg1 stability 1089:1097 biological stability 1078:1097 biological stability 1078:1097 Growth promoting activity and biological stability were analyzed by proliferation of T1165 cells. 27884519 7 4 theme amino 927:931 arg1 sequence 938:945 amino acid sequence 927:945 amino acid sequence 927:945 The structure was analyzed by amino acid sequence and composition analysis and CD-spectra. 27884519 4 5 theme secondary 574:582 arg1 structure 584:592 the predicted secondary structure 560:592 the predicted secondary structure 560:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 5 6 dep 71 657:658 arg1 to 654:655 to 654:655 to 654:655 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 4 7 theme Modified 402:409 arg1 vectors 411:417 Modified vectors 402:417 Modified vectors 402:417 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 4 8 theme predicted 564:572 arg1 structure 584:592 the predicted secondary structure 560:592 the predicted secondary structure 560:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 3 9 theme various 355:361 arg1 kinds 363:367 various kinds 355:367 various kinds of genetically modified hIL-11s 355:399 Here, we will demonstrate the creation of various kinds of genetically modified hIL-11s. 27884519 11 10 from activity 1219:1226 arg1 N-terminal 1262:1271 N-terminal 1262:1271 N-terminal 1262:1271 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 14 11 from 3NN 1560:1562 arg1 M1 1589:1590 M1 1589:1590 M1 1589:1590 Both of 3NN and OM1 with AAATPAPG on M1 associated with hIL-11R strongly. 27884519 14 12 with 3NN 1560:1562 arg1 AAATPAPG 1577:1584 AAATPAPG 1577:1584 AAATPAPG 1577:1584 Both of 3NN and OM1 with AAATPAPG on M1 associated with hIL-11R strongly. 27884519 2 13 theme cancer 293:298 arg1 progression 300:310 IL-11 related cancer progression 279:310 IL-11 related cancer progression 279:310 Whereas, development of IL-11 inhibitor is also expected for a treatment against IL-11 related cancer progression. 27884519 6 14 theme hIL-11s 873:879 arg1 characteristics 845:859 the characteristics 841:859 the characteristics of modified hIL-11s 841:879 A large scale production system was established and the characteristics of modified hIL-11s were evaluated. 27884519 3 15 theme kinds 363:367 arg1 creation 343:350 the creation 339:350 the creation of various kinds of genetically modified hIL-11s 339:399 Here, we will demonstrate the creation of various kinds of genetically modified hIL-11s. 27884519 2 16 theme related 285:291 arg1 progression 300:310 IL-11 related cancer progression 279:310 IL-11 related cancer progression 279:310 Whereas, development of IL-11 inhibitor is also expected for a treatment against IL-11 related cancer progression. 27884519 13 17 theme growth 1462:1467 arg1 activity 1469:1476 growth activity 1462:1476 growth activity 1462:1476 M1 variants lacked growth activity though they are well glycosylated and secondary structure is very stable. 27884519 6 18 theme modified 864:871 arg1 hIL-11s 873:879 modified hIL-11s 864:879 modified hIL-11s 864:879 A large scale production system was established and the characteristics of modified hIL-11s were evaluated. 27884519 5 19 theme second 669:674 arg1 loop 676:679 the second loop 665:679 the second loop (M2:114-115 aa) 665:695 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 19 theme second 669:674 arg1 aa 693:694 M2:114-115 aa 682:694 M2:114-115 aa 682:694 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 19 theme second 669:674 arg1 loop 642:645 the first loop 632:645 the first loop (M1:70 to 71 aa) 632:662 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 12 20 theme biological 1334:1343 arg1 stabilities 1345:1355 Structural and biological stabilities 1319:1355 Structural and biological stabilities of 3NN 1319:1362 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 10 21 theme N-terminal 1146:1155 arg1 proteins 1166:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins were well glycosylated and produced. 27884519 14 22 from OM1 1568:1570 arg1 M1 1589:1590 M1 1589:1590 M1 1589:1590 Both of 3NN and OM1 with AAATPAPG on M1 associated with hIL-11R strongly. 27884519 15 23 theme potent 1714:1719 arg1 agonists 1721:1728 potent agonists 1714:1728 potent agonists 1714:1728 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 4 24 theme hIL-11 495:500 arg1 region 485:490 the region 481:490 the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure 481:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 11 25 theme NASNASNAS 1240:1248 arg1 sequence 1250:1257 NASNASNAS sequence 1240:1257 NASNASNAS sequence 1240:1257 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 12 26 theme mouse 1411:1415 arg1 blood 1417:1421 mouse blood 1411:1421 mouse blood 1411:1421 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 12 27 theme Structural 1319:1328 arg1 stabilities 1345:1355 Structural and biological stabilities 1319:1355 Structural and biological stabilities of 3NN 1319:1362 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 1 28 theme myelo-suppressive 166:182 arg1 chemotherapy 184:195 myelo-suppressive chemotherapy 166:195 myelo-suppressive chemotherapy 166:195 Interleukin-11 (IL-11) has been expected as a drug on severe thrombocytopenia caused by myelo-suppressive chemotherapy. 27884519 3 29 theme modified 384:391 arg1 hIL-11s 393:399 genetically modified hIL-11s 372:399 genetically modified hIL-11s 372:399 Here, we will demonstrate the creation of various kinds of genetically modified hIL-11s. 27884519 1 30 from drug 124:127 arg1 thrombocytopenia 139:154 severe thrombocytopenia 132:154 severe thrombocytopenia caused by myelo-suppressive chemotherapy 132:195 Interleukin-11 (IL-11) has been expected as a drug on severe thrombocytopenia caused by myelo-suppressive chemotherapy. 27884519 14 31 with OM1 1568:1570 arg1 AAATPAPG 1577:1584 AAATPAPG 1577:1584 AAATPAPG 1577:1584 Both of 3NN and OM1 with AAATPAPG on M1 associated with hIL-11R strongly. 27884519 3 32 theme hIL-11s 393:399 arg1 kinds 363:367 various kinds 355:367 various kinds of genetically modified hIL-11s 355:399 Here, we will demonstrate the creation of various kinds of genetically modified hIL-11s. 27884519 13 33 gly glycosylated 1499:1510 arg1 they 1485:1488 they 1485:1488 they 1485:1488 M1 variants lacked growth activity though they are well glycosylated and secondary structure is very stable. 27884519 11 34 theme 3NN 1231:1233 arg1 activity 1219:1226 Growth activity 1212:1226 Growth activity of 3NN with NASNASNAS sequence on N-terminal 1212:1271 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 9 35 theme T1165 1133:1137 arg1 cells 1139:1143 T1165 cells 1133:1143 T1165 cells 1133:1143 Growth promoting activity and biological stability were analyzed by proliferation of T1165 cells. 27884519 7 36 theme composition 951:961 arg1 analysis 963:970 composition analysis 951:970 composition analysis 951:970 The structure was analyzed by amino acid sequence and composition analysis and CD-spectra. 27884519 9 37 theme cells 1139:1143 arg1 proliferation 1116:1128 proliferation 1116:1128 proliferation of T1165 cells 1116:1143 Growth promoting activity and biological stability were analyzed by proliferation of T1165 cells. 27884519 5 38 theme third 702:706 arg1 loop 708:711 the third loop 698:711 the third loop (M3:160-161 aa) 698:727 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 38 theme third 702:706 arg1 aa 725:726 M3:160-161 aa 714:726 M3:160-161 aa 714:726 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 38 theme third 702:706 arg1 loop 642:645 the first loop 632:645 the first loop (M1:70 to 71 aa) 632:662 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 12 39 from time 1403:1406 arg1 blood 1417:1421 mouse blood 1411:1421 mouse blood 1411:1421 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 4 40 from site 473:476 arg1 region 485:490 the region 481:490 the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure 481:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 4 41 theme O-glycosylation 457:471 arg1 site 473:476 N- or O-glycosylation site 451:476 N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure 451:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 15 42 theme M1 1664:1665 arg1 variants 1667:1674 N-terminal and M1 variants 1649:1674 variants 1667:1674 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 0 43 theme modified 47:54 arg1 variants 68:75 genetically modified human IL-11 variants 35:75 genetically modified human IL-11 variants 35:75 Production and characterization of genetically modified human IL-11 variants. 27884519 3 44 dep demonstrate 327:337 arg1 will 322:325 will 322:325 will 322:325 Here, we will demonstrate the creation of various kinds of genetically modified hIL-11s. 27884519 13 45 theme M1 1443:1444 arg1 variants 1446:1453 M1 variants 1443:1453 M1 variants 1443:1453 M1 variants lacked growth activity though they are well glycosylated and secondary structure is very stable. 27884519 11 46 dep tenfold 1283:1289 arg1 higher 1291:1296 higher 1291:1296 higher 1291:1296 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 5 47 dep 23 624:625 arg1 to 621:622 to 621:622 to 621:622 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 6 48 theme production 803:812 arg1 system 814:819 A large scale production system 789:819 A large scale production system 789:819 A large scale production system was established and the characteristics of modified hIL-11s were evaluated. 27884519 0 49 theme IL-11 62:66 arg1 variants 68:75 genetically modified human IL-11 variants 35:75 genetically modified human IL-11 variants 35:75 Production and characterization of genetically modified human IL-11 variants. 27884519 5 50 theme first 636:640 arg1 N-terminal 595:604 N-terminal 595:604 N-terminal 595:604 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 50 theme first 636:640 arg1 loop 708:711 the third loop 698:711 the third loop (M3:160-161 aa) 698:727 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 50 theme first 636:640 arg1 aa 660:661 M1:70 to 71 aa 648:661 M1:70 to 71 aa 648:661 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 50 theme first 636:640 arg1 loop 642:645 the first loop 632:645 the first loop (M1:70 to 71 aa) 632:662 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 50 theme first 636:640 arg1 loop 676:679 the second loop 665:679 the second loop (M2:114-115 aa) 665:695 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 8 51 theme composition 1026:1036 arg1 analysis 1038:1045 monosaccharide composition analysis 1011:1045 monosaccharide composition analysis 1011:1045 Glycan was assessed by monosaccharide composition analysis. 27884519 2 52 theme inhibitor 228:236 arg1 development 207:217 development 207:217 development of IL-11 inhibitor 207:236 Whereas, development of IL-11 inhibitor is also expected for a treatment against IL-11 related cancer progression. 27884519 12 53 theme 3NN 1360:1362 arg1 stabilities 1345:1355 Structural and biological stabilities 1319:1355 Structural and biological stabilities of 3NN 1319:1362 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 15 54 theme N-terminal 1649:1658 arg1 variants 1667:1674 N-terminal and M1 variants 1649:1674 variants 1667:1674 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 0 55 theme human 56:60 arg1 variants 68:75 genetically modified human IL-11 variants 35:75 genetically modified human IL-11 variants 35:75 Production and characterization of genetically modified human IL-11 variants. 27884519 4 56 theme α-helical 535:543 arg1 motif 545:549 the core α-helical motif 526:549 the core α-helical motif based on the predicted secondary structure 526:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 7 57 theme acid 933:936 arg1 sequence 938:945 amino acid sequence 927:945 amino acid sequence 927:945 The structure was analyzed by amino acid sequence and composition analysis and CD-spectra. 27884519 15 58 theme practical 1697:1705 arg1 use 1707:1709 practical use 1697:1709 practical use as potent agonists or antagonists of hIL-11 1697:1753 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 2 59 theme IL-11 222:226 arg1 inhibitor 228:236 IL-11 inhibitor 222:236 IL-11 inhibitor 222:236 Whereas, development of IL-11 inhibitor is also expected for a treatment against IL-11 related cancer progression. 27884519 11 60 with 3NN 1231:1233 arg1 sequence 1250:1257 NASNASNAS sequence 1240:1257 NASNASNAS sequence 1240:1257 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 5 61 dep N 607:607 arg1 aa 627:628 between 22 to 23 aa 610:628 N: between 22 to 23 aa 607:628 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 4 62 theme core 530:533 arg1 motif 545:549 the core α-helical motif 526:549 the core α-helical motif based on the predicted secondary structure 526:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 8 63 theme monosaccharide 1011:1024 arg1 analysis 1038:1045 monosaccharide composition analysis 1011:1045 monosaccharide composition analysis 1011:1045 Glycan was assessed by monosaccharide composition analysis. 27884519 4 64 gly O-glycosylation 457:471 arg2 site 473:476 N- or O-glycosylation site 451:476 N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure 451:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 13 65 theme secondary 1516:1524 arg1 structure 1526:1534 secondary structure 1516:1534 secondary structure 1516:1534 M1 variants lacked growth activity though they are well glycosylated and secondary structure is very stable. 27884519 4 66 theme N- 451:452 arg1 site 473:476 N- or O-glycosylation site 451:476 N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure 451:592 Modified vectors were constructed by introducing N- or O-glycosylation site on the region of hIL-11 that does not belong to the core α-helical motif based on the predicted secondary structure. 27884519 5 67 dep C 745:745 arg1 aa 753:754 200- aa 748:754 C: 200- aa 745:754 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 5 68 dep N-terminal 595:604 arg1 N 607:607 N 607:607 N: between 22 to 23 aa 607:628 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 15 69 theme hIL-11 1748:1753 arg1 agonists 1721:1728 potent agonists 1714:1728 potent agonists 1714:1728 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 15 69 theme hIL-11 1748:1753 arg1 antagonists 1733:1743 antagonists 1733:1743 antagonists 1733:1743 These results indicate N-terminal and M1 variants will be expected for practical use as potent agonists or antagonists of hIL-11. 27884519 12 70 theme residence 1393:1401 arg1 time 1403:1406 residence time 1393:1406 residence time in mouse blood 1393:1421 Structural and biological stabilities of 3NN were also better than WT and residence time in mouse blood was longer than WT. 27884519 11 71 theme Growth 1212:1217 arg1 activity 1219:1226 Growth activity 1212:1226 Growth activity of 3NN with NASNASNAS sequence on N-terminal 1212:1271 Growth activity of 3NN with NASNASNAS sequence on N-terminal was about tenfold higher than wild type (WT). 27884519 10 72 theme modified 1157:1164 arg1 proteins 1166:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins 1146:1173 N-terminal modified proteins were well glycosylated and produced. 27884519 0 73 theme variants 68:75 arg1 characterization 15:30 characterization 15:30 characterization 15:30 Production and characterization of genetically modified human IL-11 variants. 27884519 0 73 theme variants 68:75 arg1 Production 0:9 Production 0:9 Production 0:9 Production and characterization of genetically modified human IL-11 variants. 27884519 6 74 theme scale 797:801 arg1 system 814:819 A large scale production system 789:819 A large scale production system 789:819 A large scale production system was established and the characteristics of modified hIL-11s were evaluated. 27884519 5 75 dep C-terminal 733:742 arg1 C 745:745 C 745:745 C: 200- aa 745:754 N-terminal (N: between 22 to 23 aa), the first loop (M1:70 to 71 aa), the second loop (M2:114-115 aa), the third loop (M3:160-161 aa) and C-terminal (C: 200- aa) were selected for modification. 27884519 1 76 theme severe 132:137 arg1 thrombocytopenia 139:154 severe thrombocytopenia 132:154 severe thrombocytopenia caused by myelo-suppressive chemotherapy 132:195 Interleukin-11 (IL-11) has been expected as a drug on severe thrombocytopenia caused by myelo-suppressive chemotherapy. 26869352 11 0 from type 1876:1879 arg1 different 1851:1859 different 1851:1859 different 1851:1859 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 11 0 from type 1876:1879 arg1 tissue/cell 1802:1812 the original tissue/cell 1789:1812 the original tissue/cell producing fucosylated haptoglobin 1789:1846 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 1 1 theme cancer 305:310 arg1 proteins 275:282 fucosylated proteins 263:282 fucosylated proteins 263:282 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 1 1 theme cancer 305:310 arg1 biomarkers 312:321 cancer biomarkers 305:321 cancer biomarkers 305:321 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 6 2 theme difucosylated 1169:1181 arg1 N-glycans 1183:1191 difucosylated N-glycans 1169:1191 difucosylated N-glycans 1169:1191 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 0 3 theme cancer 180:185 arg1 diagnosis 167:175 the differential diagnosis 150:175 the differential diagnosis of cancer 150:185 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 3 4 from increase 512:519 arg1 haptoglobin 536:546 fucosylated haptoglobin 524:546 fucosylated haptoglobin 524:546 However, an increase in fucosylated haptoglobin has been reported in various types of cancer. 26869352 11 5 from different 1851:1859 arg1 type 1876:1879 each cancer type 1864:1879 each cancer type 1864:1879 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 4 6 dep cancer 807:812 arg1 gallbladder 843:853 gallbladder 843:853 gallbladder 843:853 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 6 dep cancer 807:812 arg1 colon 836:840 colon 836:840 colon 836:840 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 6 dep cancer 807:812 arg1 gastric 827:833 gastric 827:833 gastric 827:833 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 6 dep cancer 807:812 arg1 pancreatic 856:865 pancreatic 856:865 pancreatic 856:865 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 6 dep cancer 807:812 arg1 esophageal 815:824 esophageal 815:824 esophageal 815:824 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 2 7 with patients 405:412 arg1 cancer 430:435 pancreatic cancer 419:435 pancreatic cancer 419:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 4 8 theme normal 924:929 arg1 controls 931:938 normal controls 924:938 normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 924:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 6 9 gly difucosylated 1169:1181 arg1 N-glycans 1183:1191 difucosylated N-glycans 1169:1191 difucosylated N-glycans 1169:1191 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 4 10 with patients 750:757 arg1 types 769:773 five types 764:773 five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 764:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 1 11 theme important 207:215 arg1 type 217:220 an important type 204:220 an important type of glycosylation involved in cancer 204:256 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 1 11 theme important 207:215 arg1 Fucosylation 188:199 Fucosylation 188:199 Fucosylation 188:199 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 4 12 theme cancer 895:900 arg1 types 769:773 five types 764:773 five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 764:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 0 13 from haptoglobin 63:73 arg1 sera 78:81 sera 78:81 sera of patients with various types of cancer 78:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 3 14 theme cancer 586:591 arg1 types 577:581 various types 569:581 various types of cancer 569:591 However, an increase in fucosylated haptoglobin has been reported in various types of cancer. 26869352 4 15 theme prostate 903:910 arg1 cancer 895:900 a non-gastroenterological cancer 869:900 a non-gastroenterological cancer (prostate cancer) 869:918 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 15 theme prostate 903:910 arg1 cancer 912:917 prostate cancer 903:917 prostate cancer 903:917 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 16 located observed 641:648 arg2 fucosylation 625:636 characteristic fucosylation 610:636 characteristic fucosylation 610:636 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 16 located observed 641:648 arg1 type 665:668 each cancer type 653:668 each cancer type 653:668 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 11 17 theme original 1793:1800 arg1 different 1851:1859 different 1851:1859 different 1851:1859 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 11 17 theme original 1793:1800 arg1 tissue/cell 1802:1812 the original tissue/cell 1789:1812 the original tissue/cell producing fucosylated haptoglobin 1789:1846 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 0 18 dep implication 134:144 arg1 analyses 26:33 Site-specific and linkage analyses 0:33 analyses 26:33 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 8 19 theme fucosylation 1474:1485 arg1 linkage 1463:1469 the linkage 1459:1469 the linkage of fucosylation 1459:1485 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 4 20 theme site-specific 684:696 arg1 analyses 698:705 site-specific analyses 684:705 site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 684:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 5 21 gly glycosylation 1138:1150 arg2 sites 1152:1156 all glycosylation sites 1134:1156 all glycosylation sites 1134:1156 In all cancer samples, monofucosylated N-glycans were significantly increased at all glycosylation sites. 26869352 6 22 theme cancer 1245:1250 arg1 samples 1252:1258 cancer samples 1245:1258 cancer samples 1245:1258 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 0 23 theme patients 86:93 arg1 sera 78:81 sera 78:81 sera of patients with various types of cancer 78:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 5 24 theme cancer 1060:1065 arg1 samples 1067:1073 all cancer samples 1056:1073 all cancer samples 1056:1073 In all cancer samples, monofucosylated N-glycans were significantly increased at all glycosylation sites. 26869352 9 25 theme cancer 1573:1578 arg1 samples 1580:1586 gastroenterological cancer samples 1553:1586 gastroenterological cancer samples 1553:1586 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 9 26 theme fucosylated 1513:1523 arg1 N-glycans 1525:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 27 theme column 950:955 arg1 LC-ESI 957:962 ODS column LC-ESI 946:962 ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 946:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 10 28 theme Lewis-type 1713:1722 arg1 N-glycan 1736:1743 Lewis-type fucosylated N-glycan 1713:1743 Lewis-type fucosylated N-glycan 1713:1743 In metastatic prostate cancer, Lewis-type fucosylated N-glycan was also increased. 26869352 11 29 gly fucosylated 1824:1834 arg1 haptoglobin 1836:1846 fucosylated haptoglobin 1824:1846 fucosylated haptoglobin 1824:1846 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 10 30 theme prostate 1696:1703 arg1 cancer 1705:1710 metastatic prostate cancer 1685:1710 metastatic prostate cancer 1685:1710 In metastatic prostate cancer, Lewis-type fucosylated N-glycan was also increased. 26869352 0 31 theme various 100:106 arg1 types 108:112 various types 100:112 various types of cancer 100:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 2 32 theme mass 472:475 arg1 analyses 490:497 mass spectrometry analyses 472:497 mass spectrometry analyses 472:497 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 0 33 theme Site-specific 0:12 arg1 analyses 26:33 Site-specific and linkage analyses 0:33 analyses 26:33 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 1 34 theme fucosylated 263:273 arg1 proteins 275:282 fucosylated proteins 263:282 fucosylated proteins 263:282 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 1 34 theme fucosylated 263:273 arg1 biomarkers 312:321 cancer biomarkers 305:321 cancer biomarkers 305:321 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 7 35 theme cancer 1312:1317 arg1 types 1319:1323 cancer types 1312:1323 cancer types 1312:1323 Remarkable differences in N-glycan structure among cancer types were not observed. 26869352 2 36 gly fucosylated 353:363 arg1 N-glycans 365:373 fucosylated N-glycans 353:373 fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer 353:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 4 37 theme patients 750:757 arg1 sera 742:745 the sera 738:745 the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 738:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 10 38 theme fucosylated 1724:1734 arg1 N-glycan 1736:1743 Lewis-type fucosylated N-glycan 1713:1743 Lewis-type fucosylated N-glycan 1713:1743 In metastatic prostate cancer, Lewis-type fucosylated N-glycan was also increased. 26869352 0 39 theme linkage 18:24 arg1 analyses 26:33 Site-specific and linkage analyses 0:33 analyses 26:33 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 9 40 theme fucosylated 1608:1618 arg1 N-glycan 1620:1627 only core-type fucosylated N-glycan 1593:1627 only core-type fucosylated N-glycan 1593:1627 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 41 theme potential 989:997 arg1 sites 1013:1017 four potential glycosylation sites 984:1017 four potential glycosylation sites 984:1017 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 11 42 theme differential 1963:1974 arg1 diagnosis 1976:1984 a differential diagnosis 1961:1984 a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers 1961:2056 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 2 43 from haptoglobin 378:388 arg1 sera 397:400 the sera 393:400 the sera of patients with pancreatic cancer 393:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 0 44 theme N-glycans 50:58 arg1 analyses 26:33 Site-specific and linkage analyses 0:33 analyses 26:33 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 5 45 theme glycosylation 1138:1150 arg1 sites 1152:1156 all glycosylation sites 1134:1156 all glycosylation sites 1134:1156 In all cancer samples, monofucosylated N-glycans were significantly increased at all glycosylation sites. 26869352 7 46 theme Remarkable 1261:1270 arg1 differences 1272:1282 Remarkable differences 1261:1282 Remarkable differences in N-glycan structure among cancer types 1261:1323 Remarkable differences in N-glycan structure among cancer types were not observed. 26869352 8 47 theme carbon 1423:1428 arg1 column 1430:1435 graphitized carbon column 1411:1435 graphitized carbon column 1411:1435 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 11 48 theme fucosylation 1896:1907 arg1 linkage 1885:1891 linkage 1885:1891 linkage of fucosylation 1885:1907 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 11 48 theme fucosylation 1896:1907 arg1 clue 1920:1923 a clue 1918:1923 a clue of primary lesion 1918:1941 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 4 49 from haptoglobin 723:733 arg1 sera 742:745 the sera 738:745 the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 738:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 9 50 theme prostate 1657:1664 arg1 samples 1673:1679 prostate cancer samples 1657:1679 prostate cancer samples 1657:1679 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 8 51 theme LC-ESI 1437:1442 arg1 MS 1444:1445 LC-ESI MS 1437:1445 LC-ESI MS 1437:1445 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 11 52 theme primary 1928:1934 arg1 lesion 1936:1941 primary lesion 1928:1941 primary lesion 1928:1941 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 1 53 theme glycosylation 225:237 arg1 type 217:220 an important type 204:220 an important type of glycosylation involved in cancer 204:256 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 1 53 theme glycosylation 225:237 arg1 Fucosylation 188:199 Fucosylation 188:199 Fucosylation 188:199 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 4 54 theme characteristic 610:623 arg1 fucosylation 625:636 characteristic fucosylation 610:636 characteristic fucosylation 610:636 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 55 from analyses 698:705 arg1 haptoglobin 723:733 haptoglobin 723:733 haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 723:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 3 56 theme fucosylated 524:534 arg1 haptoglobin 536:546 fucosylated haptoglobin 524:546 fucosylated haptoglobin 524:546 However, an increase in fucosylated haptoglobin has been reported in various types of cancer. 26869352 4 57 theme cancer 658:663 arg1 type 665:668 each cancer type 653:668 each cancer type 653:668 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 8 58 theme graphitized 1411:1421 arg1 column 1430:1435 graphitized carbon column 1411:1435 graphitized carbon column 1411:1435 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 4 59 dep Asn184 1020:1025 arg1 Asn241 1044:1049 Asn241 1044:1049 Asn241 1044:1049 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 59 dep Asn184 1020:1025 arg1 Asn211 1036:1041 Asn211 1036:1041 Asn211 1036:1041 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 3 60 theme various 569:575 arg1 types 577:581 various types 569:581 various types of cancer 569:591 However, an increase in fucosylated haptoglobin has been reported in various types of cancer. 26869352 2 61 from N-glycans 365:373 arg1 haptoglobin 378:388 haptoglobin 378:388 haptoglobin in the sera of patients with pancreatic cancer 378:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 11 62 theme cancer 1869:1874 arg1 type 1876:1879 each cancer type 1864:1879 each cancer type 1864:1879 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 0 63 theme cancer 117:122 arg1 types 108:112 various types 100:112 various types of cancer 100:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 0 64 with patients 86:93 arg1 types 108:112 various types 100:112 various types of cancer 100:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 5 65 gly monofucosylated 1076:1090 arg1 N-glycans 1092:1100 monofucosylated N-glycans 1076:1100 monofucosylated N-glycans 1076:1100 In all cancer samples, monofucosylated N-glycans were significantly increased at all glycosylation sites. 26869352 0 66 theme possible 125:132 arg1 implication 134:144 possible implication 125:144 possible implication for the differential diagnosis of cancer 125:185 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 10 67 gly fucosylated 1724:1734 arg1 N-glycan 1736:1743 Lewis-type fucosylated N-glycan 1713:1743 Lewis-type fucosylated N-glycan 1713:1743 In metastatic prostate cancer, Lewis-type fucosylated N-glycan was also increased. 26869352 11 68 theme non-gastroenterological 2026:2048 arg1 cancers 2050:2056 non-gastroenterological cancers 2026:2056 non-gastroenterological cancers 2026:2056 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 4 69 dep has 980:982 arg1 Asn184 1020:1025 Asn184 1020:1025 Asn184 1020:1025 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 1 70 gly fucosylated 263:273 arg1 proteins 275:282 fucosylated proteins 263:282 fucosylated proteins 263:282 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 1 70 gly fucosylated 263:273 arg1 biomarkers 312:321 cancer biomarkers 305:321 cancer biomarkers 305:321 Fucosylation is an important type of glycosylation involved in cancer, and fucosylated proteins could be employed as cancer biomarkers. 26869352 9 71 theme Lewis-type 1488:1497 arg1 N-glycans 1525:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 72 theme non-gastroenterological 871:893 arg1 cancer 895:900 a non-gastroenterological cancer 869:900 a non-gastroenterological cancer (prostate cancer) 869:918 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 72 theme non-gastroenterological 871:893 arg1 cancer 912:917 prostate cancer 903:917 prostate cancer 903:917 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 73 theme N-glycans 710:718 arg1 analyses 698:705 site-specific analyses 684:705 site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 684:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 0 74 theme differential 154:165 arg1 diagnosis 167:175 the differential diagnosis 150:175 the differential diagnosis of cancer 150:185 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 4 75 theme gastroenterological 787:805 arg1 cancer 807:812 operable gastroenterological cancer 778:812 operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic) 778:866 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 9 76 theme core-type 1503:1511 arg1 N-glycans 1525:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 0 77 from analyses 26:33 arg1 haptoglobin 63:73 haptoglobin 63:73 haptoglobin in sera of patients with various types of cancer 63:122 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 2 78 theme spectrometry 477:488 arg1 analyses 490:497 mass spectrometry analyses 472:497 mass spectrometry analyses 472:497 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 9 79 gly fucosylated 1513:1523 arg1 N-glycans 1525:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans 1488:1533 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 3 80 gly fucosylated 524:534 arg1 haptoglobin 536:546 fucosylated haptoglobin 524:546 fucosylated haptoglobin 524:546 However, an increase in fucosylated haptoglobin has been reported in various types of cancer. 26869352 0 81 gly fucosylated 38:48 arg1 N-glycans 50:58 fucosylated N-glycans 38:58 fucosylated N-glycans 38:58 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 9 82 theme gastroenterological 1553:1571 arg1 samples 1580:1586 gastroenterological cancer samples 1553:1586 gastroenterological cancer samples 1553:1586 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 83 theme controls 931:938 arg1 types 769:773 five types 764:773 five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 764:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 11 84 theme fucosylated 1824:1834 arg1 haptoglobin 1836:1846 fucosylated haptoglobin 1824:1846 fucosylated haptoglobin 1824:1846 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 4 85 gly fucosylation 625:636 arg1 type 665:668 each cancer type 653:668 each cancer type 653:668 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 86 theme ODS 946:948 arg1 LC-ESI 957:962 ODS column LC-ESI 946:962 ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 946:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 10 87 theme metastatic 1685:1694 arg1 cancer 1705:1710 metastatic prostate cancer 1685:1710 metastatic prostate cancer 1685:1710 In metastatic prostate cancer, Lewis-type fucosylated N-glycan was also increased. 26869352 8 88 attach released 1379:1386 arg2 alditols 1370:1377 N-glycan alditols 1361:1377 N-glycan alditols released from haptoglobin 1361:1403 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 8 88 attach released 1379:1386 arg1 haptoglobin 1393:1403 haptoglobin 1393:1403 haptoglobin 1393:1403 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 5 89 theme monofucosylated 1076:1090 arg1 N-glycans 1092:1100 monofucosylated N-glycans 1076:1100 monofucosylated N-glycans 1076:1100 In all cancer samples, monofucosylated N-glycans were significantly increased at all glycosylation sites. 26869352 6 90 located detected 1198:1205 arg1 samples 1252:1258 cancer samples 1245:1258 cancer samples 1245:1258 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 6 90 located detected 1198:1205 arg1 Asn207 1219:1224 Asn207 1219:1224 Asn207 1219:1224 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 6 90 located detected 1198:1205 arg1 Asn241 1230:1235 Asn241 1230:1235 Asn241 1230:1235 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 6 90 located detected 1198:1205 arg1 Asn 1210:1212 Asn 184 1210:1216 Asn 184 1210:1216 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 6 90 located detected 1198:1205 arg2 N-glycans 1183:1191 difucosylated N-glycans 1169:1191 difucosylated N-glycans 1169:1191 Moreover, difucosylated N-glycans were detected at Asn 184, Asn207 and Asn241 only in cancer samples. 26869352 4 91 gly glycosylation 999:1011 arg2 four 984:987 four 984:987 four 984:987 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 91 gly glycosylation 999:1011 arg2 sites 1013:1017 four potential glycosylation sites 984:1017 four potential glycosylation sites 984:1017 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 9 92 theme core-type 1598:1606 arg1 N-glycan 1620:1627 only core-type fucosylated N-glycan 1593:1627 only core-type fucosylated N-glycan 1593:1627 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 7 93 theme N-glycan 1287:1294 arg1 structure 1296:1304 N-glycan structure 1287:1304 N-glycan structure 1287:1304 Remarkable differences in N-glycan structure among cancer types were not observed. 26869352 2 94 theme pancreatic 419:428 arg1 cancer 430:435 pancreatic cancer 419:435 pancreatic cancer 419:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 8 95 theme N-glycan 1361:1368 arg1 alditols 1370:1377 N-glycan alditols 1361:1377 N-glycan alditols released from haptoglobin 1361:1403 We next analyzed N-glycan alditols released from haptoglobin using graphitized carbon column LC-ESI MS to identify the linkage of fucosylation. 26869352 2 96 theme patients 405:412 arg1 sera 397:400 the sera 393:400 the sera of patients with pancreatic cancer 393:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26869352 0 97 theme fucosylated 38:48 arg1 N-glycans 50:58 fucosylated N-glycans 38:58 fucosylated N-glycans 38:58 Site-specific and linkage analyses of fucosylated N-glycans on haptoglobin in sera of patients with various types of cancer: possible implication for the differential diagnosis of cancer. 26869352 11 98 theme gastroenterological 1994:2012 arg1 cancers 2014:2020 gastroenterological cancers 1994:2020 gastroenterological cancers 1994:2020 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 7 99 from differences 1272:1282 arg1 structure 1296:1304 N-glycan structure 1287:1304 N-glycan structure 1287:1304 Remarkable differences in N-glycan structure among cancer types were not observed. 26869352 4 100 theme glycosylation 999:1011 arg1 sites 1013:1017 four potential glycosylation sites 984:1017 four potential glycosylation sites 984:1017 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 9 101 gly fucosylated 1608:1618 arg1 N-glycan 1620:1627 only core-type fucosylated N-glycan 1593:1627 only core-type fucosylated N-glycan 1593:1627 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 102 theme operable 778:785 arg1 cancer 807:812 operable gastroenterological cancer 778:812 operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic) 778:866 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 11 103 theme lesion 1936:1941 arg1 linkage 1885:1891 linkage 1885:1891 linkage of fucosylation 1885:1907 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 11 103 theme lesion 1936:1941 arg1 clue 1920:1923 a clue 1918:1923 a clue of primary lesion 1918:1941 These data suggest that the original tissue/cell producing fucosylated haptoglobin is different in each cancer type and linkage of fucosylation might be a clue of primary lesion, thereby enabling a differential diagnosis between gastroenterological cancers and non-gastroenterological cancers. 26869352 9 104 theme cancer 1666:1671 arg1 samples 1673:1679 prostate cancer samples 1657:1679 prostate cancer samples 1657:1679 Lewis-type and core-type fucosylated N-glycans were increased in gastroenterological cancer samples, but only core-type fucosylated N-glycan was relatively increased in prostate cancer samples. 26869352 4 105 contain has 980:982 arg2 sites 1013:1017 four potential glycosylation sites 984:1017 four potential glycosylation sites 984:1017 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 105 contain has 980:982 arg2 MS. 964:966 MS. 964:966 MS. 964:966 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 105 contain has 980:982 arg1 Haptoglobin 968:978 Haptoglobin 968:978 Haptoglobin 968:978 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 4 106 theme cancer 807:812 arg1 types 769:773 five types 764:773 five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241) 764:1050 To ascertain if characteristic fucosylation is observed in each cancer type, we undertook site-specific analyses of N-glycans on haptoglobin in the sera of patients with five types of operable gastroenterological cancer (esophageal, gastric, colon, gallbladder, pancreatic), a non-gastroenterological cancer (prostate cancer) and normal controls using ODS column LC-ESI MS. Haptoglobin has four potential glycosylation sites (Asn184, Asn207, Asn211, Asn241). 26869352 2 107 theme fucosylated 353:363 arg1 N-glycans 365:373 fucosylated N-glycans 353:373 fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer 353:435 Previously, we reported that fucosylated N-glycans on haptoglobin in the sera of patients with pancreatic cancer were increased by lectin-ELISA and mass spectrometry analyses. 26582203 5 0 theme cleavage 813:820 arg1 sites 822:826 protease cleavage sites 804:826 protease cleavage sites 804:826 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 3 1 contain carries 525:531 arg2 glycan 540:545 a core glycan 533:545 a core glycan 533:545 Mass spectrometry revealed that Asn-95 carries a core glycan, consisting of two GlcNAc and three hexoses. 26582203 3 1 contain carries 525:531 arg1 Asn-95 518:523 Asn-95 518:523 Asn-95 518:523 Mass spectrometry revealed that Asn-95 carries a core glycan, consisting of two GlcNAc and three hexoses. 26582203 5 2 theme specificity 733:743 arg1 similar 840:846 similar 840:846 similar 840:846 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 5 2 theme specificity 733:743 arg1 patterns 745:752 The specificity patterns 729:752 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method 729:834 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 4 3 theme Autocatalytic 592:604 arg1 activation 606:615 Autocatalytic activation 592:615 Autocatalytic activation 592:615 Autocatalytic activation was retarded in glyco-pro-KLK2, whereas the activated glyco-form exhibited an increased proteolytic resistance. 26582203 5 4 theme sites 822:826 arg1 identification 786:799 proteomic identification 776:799 proteomic identification of protease cleavage sites 776:826 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 6 5 theme enzymatic 943:951 arg1 activity 953:960 the enzymatic activity 939:960 the enzymatic activity of KLK2 939:968 However, glycosylation changes the enzymatic activity of KLK2 in a drastically substrate-dependent manner. 26582203 1 6 theme KLK3/prostate-specific 241:262 arg1 antigen 264:270 KLK3/prostate-specific antigen 241:270 KLK3/prostate-specific antigen 241:270 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 5 7 theme PICS 770:773 arg1 method 829:834 the PICS (proteomic identification of protease cleavage sites) method 766:834 the PICS (proteomic identification of protease cleavage sites) method 766:834 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 9 8 dep favor 1395:1399 arg1 i.e. 1492:1495 i.e. 1492:1495 i.e. 1492:1495 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 10 9 from kinetics 1551:1558 arg1 inactivation 1573:1584 autolytic inactivation 1563:1584 autolytic inactivation of both KLK2 variants 1563:1606 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 10 10 theme variants 1599:1606 arg1 inactivation 1573:1584 autolytic inactivation 1563:1584 autolytic inactivation of both KLK2 variants 1563:1606 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 2 11 from expression 474:483 arg1 counterpart 440:450 its non-glycosylated counterpart 419:450 its non-glycosylated counterpart from Escherichia coli expression 419:483 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 7 12 theme protein 1177:1183 arg1 glyco-pro-KLK2 1205:1218 glyco-pro-KLK2 itself 1205:1225 glyco-pro-KLK2 itself 1205:1225 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 7 12 theme protein 1177:1183 arg1 substrates 1185:1194 protein substrates 1177:1194 protein substrates 1177:1194 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 10 13 theme due 1656:1658 arg1 sites 1650:1654 the target sites 1639:1654 the target sites due to the presence of the glycan 1639:1688 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 5 14 with similar 840:846 arg1 preference 885:894 a major preference 877:894 a major preference for P1-Arg 877:905 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 5 15 theme proteomic 776:784 arg1 identification 786:799 proteomic identification 776:799 proteomic identification of protease cleavage sites 776:826 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 9 16 theme non-glycosylated 1361:1376 arg1 99-loop 1378:1384 the non-glycosylated 99-loop 1357:1384 the non-glycosylated 99-loop 1357:1384 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 8 17 theme active 1320:1325 arg1 site 1327:1330 the active site 1316:1330 the active site 1316:1330 These findings can be rationalized by the glycan-carrying 99-loop that prefers to cover the active site like a lid. 26582203 2 18 theme tarentolae 381:390 arg1 cells 392:396 Leishmania tarentolae cells 370:396 Leishmania tarentolae cells 370:396 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 0 19 theme Enzymatic 96:104 arg1 Activity 106:113 Enzymatic Activity 96:113 Enzymatic Activity 96:113 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 0 20 from 99-Loop 23:29 arg1 Glycan 9:14 A Single Glycan 0:14 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 0:69 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 10 21 from pattern 1539:1545 arg1 inactivation 1573:1584 autolytic inactivation 1563:1584 autolytic inactivation of both KLK2 variants 1563:1606 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 1 22 theme prostate 211:218 arg1 cancer 220:225 prostate cancer 211:225 prostate cancer 211:225 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 10 23 theme sites 1650:1654 arg1 shift 1630:1634 a shift 1628:1634 a shift of the target sites due to the presence of the glycan 1628:1688 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 10 24 theme KLK2 1594:1597 arg1 variants 1599:1606 both KLK2 variants 1589:1606 both KLK2 variants 1589:1606 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 0 25 theme Single 2:7 arg1 Glycan 9:14 A Single Glycan 0:14 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 0:69 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 6 26 theme substrate-dependent 987:1005 arg1 manner 1007:1012 a drastically substrate-dependent manner 973:1012 a drastically substrate-dependent manner 973:1012 However, glycosylation changes the enzymatic activity of KLK2 in a drastically substrate-dependent manner. 26582203 2 27 dep Escherichia 457:467 arg1 coli 469:472 coli 469:472 coli 469:472 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 7 28 contain has 1035:1037 arg1 glyco-KLK2 1024:1033 glyco-KLK2 1024:1033 glyco-KLK2 1024:1033 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 7 28 contain has 1035:1037 arg2 efficiency 1070:1079 a considerably lower catalytic efficiency 1039:1079 a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe 1039:1140 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 10 29 theme cleavage 1530:1537 arg1 pattern 1539:1545 cleavage pattern 1530:1545 cleavage pattern 1530:1545 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 1 30 theme key 165:167 arg1 peptidase 141:149 Human kallikrein-related peptidase 2 116:151 Human kallikrein-related peptidase 2 (KLK2) 116:158 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 30 theme key 165:167 arg1 protease 176:183 a key serine protease 163:183 a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen 163:270 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 11 31 theme kallikrein-related 1773:1790 arg1 biology 1802:1808 kallikrein-related peptidase biology 1773:1808 kallikrein-related peptidase biology 1773:1808 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 11 32 theme pathology 1814:1822 arg1 understanding 1756:1768 a deeper understanding 1747:1768 a deeper understanding of kallikrein-related peptidase biology and pathology 1747:1822 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 11 33 theme glycosylation 1717:1729 arg1 effects 1706:1712 These striking effects 1691:1712 These striking effects of glycosylation 1691:1729 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 1 34 theme serine 169:174 arg1 peptidase 141:149 Human kallikrein-related peptidase 2 116:151 Human kallikrein-related peptidase 2 (KLK2) 116:158 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 34 theme serine 169:174 arg1 protease 176:183 a key serine protease 163:183 a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen 163:270 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 5 35 theme KLK2 857:860 arg1 variants 862:869 both KLK2 variants 852:869 both KLK2 variants 852:869 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 11 36 theme biology 1802:1808 arg1 understanding 1756:1768 a deeper understanding 1747:1768 a deeper understanding of kallikrein-related peptidase biology and pathology 1747:1822 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 2 37 from pro-KLK2 358:365 arg1 cells 392:396 Leishmania tarentolae cells 370:396 Leishmania tarentolae cells 370:396 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 9 38 gly non-glycosylated 1361:1376 arg1 99-loop 1378:1384 the non-glycosylated 99-loop 1357:1384 the non-glycosylated 99-loop 1357:1384 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 7 39 with substrates 1119:1128 arg1 P2-Phe 1135:1140 P2-Phe 1135:1140 P2-Phe 1135:1140 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 2 40 theme non-glycosylated 423:438 arg1 counterpart 440:450 its non-glycosylated counterpart 419:450 its non-glycosylated counterpart from Escherichia coli expression 419:483 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 10 41 dep pattern 1539:1545 arg1 the 1526:1528 the 1526:1528 the 1526:1528 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 7 42 theme lower 1054:1058 arg1 efficiency 1070:1079 a considerably lower catalytic efficiency 1039:1079 a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe 1039:1140 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 11 43 theme striking 1697:1704 arg1 effects 1706:1712 These striking effects 1691:1712 These striking effects of glycosylation 1691:1729 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 3 44 theme Mass 486:489 arg1 spectrometry 491:502 Mass spectrometry 486:502 Mass spectrometry 486:502 Mass spectrometry revealed that Asn-95 carries a core glycan, consisting of two GlcNAc and three hexoses. 26582203 10 45 theme target 1643:1648 arg1 sites 1650:1654 the target sites 1639:1654 the target sites due to the presence of the glycan 1639:1688 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 2 46 theme site 340:343 arg1 function 298:305 the function 294:305 the function of its potential N-glycosylation site 294:343 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 9 47 theme Km 1515:1516 arg1 reduction 1502:1510 a reduction 1500:1510 a reduction of Km 1500:1516 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 4 48 theme increased 695:703 arg1 resistance 717:726 an increased proteolytic resistance 692:726 an increased proteolytic resistance 692:726 Autocatalytic activation was retarded in glyco-pro-KLK2, whereas the activated glyco-form exhibited an increased proteolytic resistance. 26582203 1 49 theme semen 188:192 arg1 liquefaction 194:205 semen liquefaction 188:205 semen liquefaction 188:205 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 8 50 theme glycan-carrying 1270:1284 arg1 99-loop 1286:1292 the glycan-carrying 99-loop 1266:1292 the glycan-carrying 99-loop that prefers to cover the active site like a lid 1266:1341 These findings can be rationalized by the glycan-carrying 99-loop that prefers to cover the active site like a lid. 26582203 9 51 theme open 1408:1411 arg1 conformation 1413:1424 a wide open conformation 1401:1424 a wide open conformation 1401:1424 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 11 52 theme peptidase 1792:1800 arg1 biology 1802:1808 kallikrein-related peptidase biology 1773:1808 kallikrein-related peptidase biology 1773:1808 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 11 53 theme deeper 1749:1754 arg1 understanding 1756:1768 a deeper understanding 1747:1768 a deeper understanding of kallikrein-related peptidase biology and pathology 1747:1822 These striking effects of glycosylation pave the way to a deeper understanding of kallikrein-related peptidase biology and pathology. 26582203 4 54 theme activated 661:669 arg1 glyco-form 671:680 the activated glyco-form 657:680 the activated glyco-form 657:680 Autocatalytic activation was retarded in glyco-pro-KLK2, whereas the activated glyco-form exhibited an increased proteolytic resistance. 26582203 0 55 theme Human 34:38 arg1 Peptidase 59:67 Human Kallikrein-related Peptidase 2 34:69 Human Kallikrein-related Peptidase 2 34:69 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 4 56 theme proteolytic 705:715 arg1 resistance 717:726 an increased proteolytic resistance 692:726 an increased proteolytic resistance 692:726 Autocatalytic activation was retarded in glyco-pro-KLK2, whereas the activated glyco-form exhibited an increased proteolytic resistance. 26582203 5 57 theme protease 804:811 arg1 sites 822:826 protease cleavage sites 804:826 protease cleavage sites 804:826 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 7 58 theme glycan-free 1086:1096 arg1 KLK2 1098:1101 glycan-free KLK2 1086:1101 glycan-free KLK2 1086:1101 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 7 59 theme peptidic 1110:1117 arg1 substrates 1119:1128 peptidic substrates 1110:1128 peptidic substrates with P2-Phe 1110:1140 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 10 60 theme autolytic 1563:1571 arg1 inactivation 1573:1584 autolytic inactivation 1563:1584 autolytic inactivation of both KLK2 variants 1563:1606 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 1 61 from protease 176:183 arg1 cancer 220:225 prostate cancer 211:225 prostate cancer 211:225 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 61 from protease 176:183 arg1 liquefaction 194:205 semen liquefaction 188:205 semen liquefaction 188:205 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 62 theme Human 116:120 arg1 peptidase 141:149 Human kallikrein-related peptidase 2 116:151 Human kallikrein-related peptidase 2 (KLK2) 116:158 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 62 theme Human 116:120 arg1 KLK2 154:157 KLK2 154:157 KLK2 154:157 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 62 theme Human 116:120 arg1 protease 176:183 a key serine protease 163:183 a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen 163:270 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 6 63 theme KLK2 965:968 arg1 activity 953:960 the enzymatic activity 939:960 the enzymatic activity of KLK2 939:968 However, glycosylation changes the enzymatic activity of KLK2 in a drastically substrate-dependent manner. 26582203 5 64 dep PICS 770:773 arg1 identification 786:799 proteomic identification 776:799 proteomic identification of protease cleavage sites 776:826 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 9 65 theme wide 1403:1406 arg1 conformation 1413:1424 a wide open conformation 1401:1424 a wide open conformation 1401:1424 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 26582203 7 66 theme catalytic 1060:1068 arg1 efficiency 1070:1079 a considerably lower catalytic efficiency 1039:1079 a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe 1039:1140 Although glyco-KLK2 has a considerably lower catalytic efficiency than glycan-free KLK2 toward peptidic substrates with P2-Phe, the situation was reverted toward protein substrates, such as glyco-pro-KLK2 itself. 26582203 1 67 theme kallikrein-related 122:139 arg1 peptidase 141:149 Human kallikrein-related peptidase 2 116:151 Human kallikrein-related peptidase 2 (KLK2) 116:158 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 67 theme kallikrein-related 122:139 arg1 KLK2 154:157 KLK2 154:157 KLK2 154:157 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 1 67 theme kallikrein-related 122:139 arg1 protease 176:183 a key serine protease 163:183 a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen 163:270 Human kallikrein-related peptidase 2 (KLK2) is a key serine protease in semen liquefaction and prostate cancer together with KLK3/prostate-specific antigen. 26582203 2 68 gly N-glycosylation 324:338 arg2 site 340:343 its potential N-glycosylation site 310:343 its potential N-glycosylation site 310:343 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 2 69 theme potential 314:322 arg1 site 340:343 its potential N-glycosylation site 310:343 its potential N-glycosylation site 310:343 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 2 70 theme N-glycosylation 324:338 arg1 site 340:343 its potential N-glycosylation site 310:343 its potential N-glycosylation site 310:343 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 0 71 theme Peptidase 59:67 arg1 99-Loop 23:29 the 99-Loop 19:29 the 99-Loop of Human Kallikrein-related Peptidase 2 19:69 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 5 72 theme major 879:883 arg1 preference 885:894 a major preference 877:894 a major preference for P1-Arg 877:905 The specificity patterns obtained by the PICS (proteomic identification of protease cleavage sites) method are similar for both KLK2 variants, with a major preference for P1-Arg. 26582203 2 73 gly non-glycosylated 423:438 arg1 counterpart 440:450 its non-glycosylated counterpart 419:450 its non-glycosylated counterpart from Escherichia coli expression 419:483 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 10 74 theme glycan 1683:1688 arg1 presence 1667:1674 the presence 1663:1674 the presence of the glycan 1663:1688 Also, the cleavage pattern and kinetics in autolytic inactivation of both KLK2 variants can be explained by a shift of the target sites due to the presence of the glycan. 26582203 3 75 theme core 535:538 arg1 glycan 540:545 a core glycan 533:545 a core glycan 533:545 Mass spectrometry revealed that Asn-95 carries a core glycan, consisting of two GlcNAc and three hexoses. 26582203 0 76 theme Kallikrein-related 40:57 arg1 Peptidase 59:67 Human Kallikrein-related Peptidase 2 34:69 Human Kallikrein-related Peptidase 2 34:69 A Single Glycan at the 99-Loop of Human Kallikrein-related Peptidase 2 Regulates Activation and Enzymatic Activity. 26582203 2 77 theme Escherichia 457:467 arg1 expression 474:483 Escherichia coli expression 457:483 Escherichia coli expression 457:483 In order to decipher the function of its potential N-glycosylation site, we produced pro-KLK2 in Leishmania tarentolae cells and compared it with its non-glycosylated counterpart from Escherichia coli expression. 26582203 9 78 theme apparent 1454:1461 arg1 affinity 1463:1470 the apparent affinity 1450:1470 the apparent affinity for the substrates 1450:1489 By contrast, the non-glycosylated 99-loop seems to favor a wide open conformation, which mostly increases the apparent affinity for the substrates (i.e. by a reduction of Km). 27493216 2 0 theme α-dystroglycan 367:380 arg1 structure 354:362 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure 294:362 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, 294:452 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 8 1 dep findings 1761:1768 arg1 the 1744:1746 the 1744:1746 the 1744:1746 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 8 1 dep findings 1761:1768 arg1 basis 1748:1752 basis 1748:1752 basis 1748:1752 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 4 2 theme -Man 1093:1096 arg1 glycan 1098:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 5 3 theme hydrophobic 1272:1282 arg1 interactions 1284:1295 hydrophobic interactions 1272:1295 hydrophobic interactions with little sequence specificity 1272:1328 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 3 4 theme O-mannosyl 705:714 arg1 synthesis 723:731 O-mannosyl glycan synthesis 705:731 O-mannosyl glycan synthesis 705:731 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 1 5 theme basement 183:190 arg1 membrane 192:199 the basement membrane 179:199 the basement membrane 179:199 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 4 6 theme GlcNAc-β1,2-Man 880:894 arg1 glycan 896:901 GlcNAc-β1,2-Man glycan 880:901 GlcNAc-β1,2-Man glycan 880:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 5 7 theme catalytic 1206:1214 arg1 domain 1216:1221 the catalytic domain 1202:1221 the catalytic domain 1202:1221 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 7 8 dep fukutin 1632:1638 arg1 e.g. 1626:1629 e.g. 1626:1629 e.g. 1626:1629 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 6 9 theme β-linked 1390:1397 arg1 GlcNAc 1399:1404 the β-linked GlcNAc 1386:1404 the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1 1386:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 3 10 theme dystrophy 588:596 arg1 types 560:564 some types 555:564 some types of inherited muscular dystrophy 555:596 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 7 11 theme GlcNAc-β1,2-Man 1536:1550 arg1 clustering 1552:1561 GlcNAc-β1,2-Man clustering 1536:1561 GlcNAc-β1,2-Man clustering 1536:1561 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 3 12 theme inherited 569:577 arg1 dystrophy 588:596 inherited muscular dystrophy 569:596 inherited muscular dystrophy 569:596 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 8 13 theme MEB 1890:1892 arg1 patients 1894:1901 MEB patients 1890:1901 MEB patients 1890:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 6 14 theme stem 1363:1366 arg1 domain 1368:1373 the stem domain 1359:1373 the stem domain 1359:1373 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 8 15 located observed 1858:1865 arg2 glycan 1851:1856 the glycan 1847:1856 the glycan observed in POMGnT1-KO mice and MEB patients 1847:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 8 15 located observed 1858:1865 arg1 mice 1881:1884 POMGnT1-KO mice 1870:1884 POMGnT1-KO mice 1870:1884 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 8 15 located observed 1858:1865 arg1 patients 1894:1901 MEB patients 1890:1901 MEB patients 1890:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 6 16 theme POMGnT1 1452:1458 arg1 glycan 1420:1425 O-mannosyl glycan 1409:1425 O-mannosyl glycan 1409:1425 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 6 16 theme POMGnT1 1452:1458 arg1 product 1441:1447 an enzymatic product 1428:1447 an enzymatic product of POMGnT1 1428:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 1 17 theme cell 162:165 arg1 membrane 167:174 the cell membrane 158:174 the cell membrane 158:174 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 4 18 theme congenital 962:971 arg1 disease 945:951 muscle-eye-brain disease 928:951 muscle-eye-brain disease (MEB) 928:957 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 18 theme congenital 962:971 arg1 dystrophy 982:990 a congenital muscular dystrophy 960:990 a congenital muscular dystrophy 960:990 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 5 19 theme crystal 1126:1132 arg1 structures 1134:1143 Our crystal structures 1122:1143 Our crystal structures of POMGnT1 1122:1154 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 8 20 theme POMGnT1-KO 1870:1879 arg1 mice 1881:1884 POMGnT1-KO mice 1870:1884 POMGnT1-KO mice 1870:1884 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 6 21 theme glycan 1420:1425 arg1 GlcNAc 1399:1404 the β-linked GlcNAc 1386:1404 the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1 1386:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 2 22 theme cell 439:442 arg1 membrane 444:451 the cell membrane 435:451 the cell membrane 435:451 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 5 23 gly O-mannosylated 1244:1257 arg1 proteins 1259:1266 substrate O-mannosylated proteins 1234:1266 substrate O-mannosylated proteins 1234:1266 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 0 24 theme Carbohydrate-binding 0:19 arg1 region 48:53 the POMGnT1 stem region 31:53 the POMGnT1 stem region 31:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 0 24 theme Carbohydrate-binding 0:19 arg1 domain 21:26 Carbohydrate-binding domain 0:26 Carbohydrate-binding domain of the POMGnT1 stem region 0:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 1 25 theme muscle 276:281 arg1 integrity 283:291 muscle integrity 276:291 muscle integrity 276:291 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 7 26 theme -Man 1728:1731 arg1 glycan 1733:1738 the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1687:1738 the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1687:1738 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 4 27 theme O-linked 753:760 arg1 β1,2-N-acetylglucosaminyltransferase 770:805 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 745:807 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) 745:817 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 27 theme O-linked 753:760 arg1 POMGnT1 810:816 POMGnT1 810:816 POMGnT1 810:816 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 27 theme O-linked 753:760 arg1 glycosyltransferase 822:840 a glycosyltransferase 820:840 a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan 820:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 1 28 theme glycoprotein 121:132 arg1 essential 205:213 essential 205:213 essential 205:213 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 1 28 theme glycoprotein 121:132 arg1 complex 134:140 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 4 29 theme POMGnT1 1014:1020 arg1 role 1006:1009 the role 1002:1009 the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1002:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 0 30 theme O-mannosylation 65:79 arg1 α-dystroglycan 90:103 α-dystroglycan 90:103 α-dystroglycan 90:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 0 30 theme O-mannosylation 65:79 arg1 sites 81:85 O-mannosylation sites 65:85 O-mannosylation sites of α-dystroglycan 65:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 3 31 theme disease-related 639:653 arg1 mutations 655:663 many disease-related mutations 634:663 many disease-related mutations 634:663 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 2 32 theme complex 407:413 arg1 subunit 392:398 a subunit 390:398 a subunit of the complex that is anchored to the cell membrane 390:451 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 2 32 theme complex 407:413 arg1 α-dystroglycan 367:380 α-dystroglycan 367:380 α-dystroglycan (α-DG) 367:387 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 0 33 theme POMGnT1 35:41 arg1 region 48:53 the POMGnT1 stem region 31:53 the POMGnT1 stem region 31:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 4 34 theme postphosphoryl 1025:1038 arg1 modification 1040:1051 postphosphoryl modification 1025:1051 postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1025:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 3 35 with consistent 599:608 arg1 relationship 620:631 this relationship 615:631 this relationship 615:631 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 0 36 theme region 48:53 arg1 region 48:53 the POMGnT1 stem region 31:53 the POMGnT1 stem region 31:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 0 36 theme region 48:53 arg1 domain 21:26 Carbohydrate-binding domain 0:26 Carbohydrate-binding domain of the POMGnT1 stem region 0:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 4 37 mod modification 1040:1051 arg1 glycan 1098:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 37 mod modification 1040:1051 arg3 postphosphoryl 1025:1038 postphosphoryl modification 1025:1051 postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1025:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 2 38 theme -Man 349:352 arg1 structure 354:362 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure 294:362 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, 294:452 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 4 39 from role 1006:1009 arg1 modification 1040:1051 postphosphoryl modification 1025:1051 postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1025:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 5 40 theme little 1302:1307 arg1 specificity 1318:1328 little sequence specificity 1302:1328 little sequence specificity 1302:1328 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 5 41 theme O-mannosylated 1244:1257 arg1 proteins 1259:1266 substrate O-mannosylated proteins 1234:1266 substrate O-mannosylated proteins 1234:1266 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 4 42 theme glycan 896:901 arg1 formation 867:875 the formation 863:875 the formation of GlcNAc-β1,2-Man glycan 863:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 43 theme glycan 1098:1103 arg1 modification 1040:1051 postphosphoryl modification 1025:1051 postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1025:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 3 44 theme α-DG 537:540 arg1 glycosylation 520:532 Reduced glycosylation 512:532 Reduced glycosylation of α-DG 512:540 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 7 45 theme α-DG 1520:1523 arg1 α-DG 1520:1523 α-DG 1520:1523 α-DG 1520:1523 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 7 45 theme α-DG 1520:1523 arg1 site 1512:1515 a specific site 1501:1515 a specific site of α-DG 1501:1523 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 7 46 theme specific 1503:1510 arg1 α-DG 1520:1523 α-DG 1520:1523 α-DG 1520:1523 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 7 46 theme specific 1503:1510 arg1 site 1512:1515 a specific site 1501:1515 a specific site of α-DG 1501:1523 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 8 47 from deficiency 1802:1811 arg1 modification 1831:1842 postphosphoryl modification 1816:1842 postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients 1816:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 3 48 theme glycan 716:721 arg1 synthesis 723:731 O-mannosyl glycan synthesis 705:731 O-mannosyl glycan synthesis 705:731 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 7 49 theme GalNAc-β1,3-GlcNAc-β1,4- 1691:1714 arg1 glycan 1733:1738 the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1687:1738 the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1687:1738 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 6 50 theme O-mannosyl 1409:1418 arg1 glycan 1420:1425 O-mannosyl glycan 1409:1425 O-mannosyl glycan 1409:1425 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 6 50 theme O-mannosyl 1409:1418 arg1 product 1441:1447 an enzymatic product 1428:1447 an enzymatic product of POMGnT1 1428:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 2 51 from laminin 478:484 arg1 membrane 502:509 the basement membrane 489:509 the basement membrane 489:509 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 3 52 theme muscular 579:586 arg1 dystrophy 588:596 inherited muscular dystrophy 569:596 inherited muscular dystrophy 569:596 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 7 53 theme further 1663:1669 arg1 modification 1671:1682 further modification 1663:1682 further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1663:1738 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 4 54 theme muscle-eye-brain 928:943 arg1 dystrophy 982:990 a congenital muscular dystrophy 960:990 a congenital muscular dystrophy 960:990 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 54 theme muscle-eye-brain 928:943 arg1 MEB 954:956 MEB 954:956 MEB 954:956 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 54 theme muscle-eye-brain 928:943 arg1 disease 945:951 muscle-eye-brain disease 928:951 muscle-eye-brain disease (MEB) 928:957 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 8 55 theme postphosphoryl 1816:1829 arg1 modification 1831:1842 postphosphoryl modification 1816:1842 postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients 1816:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 0 56 theme α-dystroglycan 90:103 arg1 α-dystroglycan 90:103 α-dystroglycan 90:103 α-dystroglycan 90:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 0 56 theme α-dystroglycan 90:103 arg1 sites 81:85 O-mannosylation sites 65:85 O-mannosylation sites of α-dystroglycan 65:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 3 57 gly glycosylation 520:532 arg1 α-DG 537:540 α-DG 537:540 α-DG 537:540 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 6 58 theme enzymatic 1431:1439 arg1 glycan 1420:1425 O-mannosyl glycan 1409:1425 O-mannosyl glycan 1409:1425 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 6 58 theme enzymatic 1431:1439 arg1 product 1441:1447 an enzymatic product 1428:1447 an enzymatic product of POMGnT1 1428:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 5 59 theme previous 1172:1179 arg1 results 1181:1187 our previous results 1168:1187 our previous results 1168:1187 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 8 60 theme glycan 1851:1856 arg1 modification 1831:1842 postphosphoryl modification 1816:1842 postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients 1816:1901 On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the glycan observed in POMGnT1-KO mice and MEB patients. 27493216 0 61 gly O-mannosylation 65:79 arg2 α-dystroglycan 90:103 α-dystroglycan 90:103 α-dystroglycan 90:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 0 61 gly O-mannosylation 65:79 arg2 sites 81:85 O-mannosylation sites 65:85 O-mannosylation sites of α-dystroglycan 65:103 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 4 62 theme mannose 762:768 arg1 β1,2-N-acetylglucosaminyltransferase 770:805 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 745:807 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) 745:817 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 62 theme mannose 762:768 arg1 POMGnT1 810:816 POMGnT1 810:816 POMGnT1 810:816 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 62 theme mannose 762:768 arg1 glycosyltransferase 822:840 a glycosyltransferase 820:840 a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan 820:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 1 63 gly glycoprotein 121:132 arg1 glycoprotein 121:132 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 2 64 theme basement 493:500 arg1 membrane 502:509 the basement membrane 489:509 the basement membrane 489:509 An O-mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with laminin in the basement membrane. 27493216 3 65 located detected 675:682 arg1 genes 687:691 genes 687:691 genes involved in O-mannosyl glycan synthesis 687:731 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 3 65 located detected 675:682 arg2 mutations 655:663 many disease-related mutations 634:663 many disease-related mutations 634:663 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 5 66 theme POMGnT1 1148:1154 arg1 structures 1134:1143 Our crystal structures 1122:1143 Our crystal structures of POMGnT1 1122:1154 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 3 67 attach linked 545:550 arg2 glycosylation 520:532 Reduced glycosylation 512:532 Reduced glycosylation of α-DG 512:540 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 3 67 attach linked 545:550 arg1 types 560:564 some types 555:564 some types of inherited muscular dystrophy 555:596 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 1 68 theme integrity 283:291 arg1 maintenance 261:271 maintenance 261:271 maintenance of muscle integrity 261:291 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 4 69 from Defects 734:740 arg1 β1,2-N-acetylglucosaminyltransferase 770:805 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 745:807 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) 745:817 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 69 from Defects 734:740 arg1 POMGnT1 810:816 POMGnT1 810:816 POMGnT1 810:816 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 69 from Defects 734:740 arg1 glycosyltransferase 822:840 a glycosyltransferase 820:840 a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan 820:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 1 70 theme dystrophin 110:119 arg1 essential 205:213 essential 205:213 essential 205:213 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 1 70 theme dystrophin 110:119 arg1 complex 134:140 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex 106:140 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 7 71 theme glycan 1733:1738 arg1 modification 1671:1682 further modification 1663:1682 further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1663:1738 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 4 72 theme GalNAc-β1,3-GlcNAc-β1,4- 1056:1079 arg1 glycan 1098:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1056:1103 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 3 73 theme many 634:637 arg1 mutations 655:663 many disease-related mutations 634:663 many disease-related mutations 634:663 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 4 74 theme protein 745:751 arg1 β1,2-N-acetylglucosaminyltransferase 770:805 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 745:807 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) 745:817 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 74 theme protein 745:751 arg1 POMGnT1 810:816 POMGnT1 810:816 POMGnT1 810:816 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 74 theme protein 745:751 arg1 glycosyltransferase 822:840 a glycosyltransferase 820:840 a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan 820:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 0 75 theme stem 43:46 arg1 region 48:53 the POMGnT1 stem region 31:53 the POMGnT1 stem region 31:53 Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan. 27493216 5 76 theme substrate 1234:1242 arg1 proteins 1259:1266 substrate O-mannosylated proteins 1234:1266 substrate O-mannosylated proteins 1234:1266 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 1 77 theme biological 232:241 arg1 events 243:248 biological events 232:248 biological events 232:248 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 1 77 theme biological 232:241 arg1 maintenance 261:271 maintenance 261:271 maintenance of muscle integrity 261:291 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 4 78 theme muscular 973:980 arg1 disease 945:951 muscle-eye-brain disease 928:951 muscle-eye-brain disease (MEB) 928:957 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 78 theme muscular 973:980 arg1 dystrophy 982:990 a congenital muscular dystrophy 960:990 a congenital muscular dystrophy 960:990 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 7 79 theme other 1584:1588 arg1 enzymes 1590:1596 other enzymes 1584:1596 other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan 1584:1738 This interaction may recruit POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other enzymes that interact with POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan. 27493216 6 80 link β-linked 1390:1397 arg1 GlcNAc 1399:1404 the β-linked GlcNAc 1386:1404 the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1 1386:1458 Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl glycan, an enzymatic product of POMGnT1. 27493216 4 81 link O-linked 753:760 arg1 β1,2-N-acetylglucosaminyltransferase 770:805 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 745:807 protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) 745:817 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 81 link O-linked 753:760 arg1 POMGnT1 810:816 POMGnT1 810:816 POMGnT1 810:816 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 4 81 link O-linked 753:760 arg1 glycosyltransferase 822:840 a glycosyltransferase 820:840 a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan 820:901 Defects in protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1), a glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man glycan, are causally related to muscle-eye-brain disease (MEB), a congenital muscular dystrophy, although the role of POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man glycan remains elusive. 27493216 1 82 theme events 243:248 arg1 variety 221:227 a variety 219:227 a variety of biological events, including maintenance of muscle integrity 219:291 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 1 82 theme events 243:248 arg1 events 243:248 biological events 232:248 biological events 232:248 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 1 82 theme events 243:248 arg1 maintenance 261:271 maintenance 261:271 maintenance of muscle integrity 261:291 The dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. 27493216 5 83 with interactions 1284:1295 arg1 specificity 1318:1328 little sequence specificity 1302:1328 little sequence specificity 1302:1328 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 27493216 3 84 theme Reduced 512:518 arg1 glycosylation 520:532 Reduced glycosylation 512:532 Reduced glycosylation of α-DG 512:540 Reduced glycosylation of α-DG is linked to some types of inherited muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl glycan synthesis. 27493216 5 85 theme sequence 1309:1316 arg1 specificity 1318:1328 little sequence specificity 1302:1328 little sequence specificity 1302:1328 Our crystal structures of POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated proteins via hydrophobic interactions with little sequence specificity. 25824821 0 0 from glycan 9:14 arg1 IgE 19:21 IgE 19:21 IgE 19:21 A single glycan on IgE is indispensable for initiation of anaphylaxis. 25824821 7 1 link N-linked 848:855 arg1 structure 870:878 a single N-linked oligomannose structure 839:878 a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE 839:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 0 2 theme anaphylaxis 58:68 arg1 initiation 44:53 initiation 44:53 initiation of anaphylaxis 44:68 A single glycan on IgE is indispensable for initiation of anaphylaxis. 25824821 9 3 theme unappreciated 1245:1257 arg1 requirement 1273:1283 an unappreciated and essential requirement 1242:1283 an unappreciated and essential requirement of glycosylation in IgE biology 1242:1315 These results underscore an unappreciated and essential requirement of glycosylation in IgE biology. 25824821 1 4 theme allergic 134:141 arg1 diseases 143:150 allergic diseases 134:150 allergic diseases 134:150 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 7 5 theme oligomannose 857:868 arg1 structure 870:878 a single N-linked oligomannose structure 839:878 a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE 839:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 4 6 theme glycosylation 559:571 arg1 patterns 573:580 precise glycosylation patterns 551:580 precise glycosylation patterns 551:580 It is well established that precise glycosylation patterns exert profound effects on the biological activity of IgG. 25824821 4 7 theme profound 588:595 arg1 effects 597:603 profound effects 588:603 profound effects 588:603 It is well established that precise glycosylation patterns exert profound effects on the biological activity of IgG. 25824821 8 8 theme IgE 1101:1103 arg1 binding 1105:1111 IgE binding 1101:1111 IgE binding to FcεRI 1101:1120 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 9 9 theme essential 1263:1271 arg1 requirement 1273:1283 an unappreciated and essential requirement 1242:1283 an unappreciated and essential requirement of glycosylation in IgE biology 1242:1315 These results underscore an unappreciated and essential requirement of glycosylation in IgE biology. 25824821 8 10 theme Genetic 976:982 arg1 disruption 984:993 Genetic disruption 976:993 Genetic disruption of the site 976:1005 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 7 11 from structure 870:878 arg1 domain 896:901 the constant domain 3 883:903 the constant domain 3 (Cε3) of IgE 883:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 11 from structure 870:878 arg1 Cε3 906:908 Cε3 906:908 Cε3 906:908 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 11 from structure 870:878 arg1 IgE 914:916 IgE 914:916 IgE 914:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 12 theme single 841:846 arg1 structure 870:878 a single N-linked oligomannose structure 839:878 a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE 839:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 6 13 theme allergic 783:790 arg1 reactions 792:800 allergic reactions 783:800 allergic reactions 783:800 Here, we demonstrate an absolute requirement for IgE glycosylation in allergic reactions. 25824821 3 14 theme cascade 514:520 arg1 initiation 487:496 initiation 487:496 initiation of the allergic cascade 487:520 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 3 14 theme cascade 514:520 arg1 release 449:455 the release 445:455 the release of inflammatory mediators 445:481 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 7 15 theme N-linked 848:855 arg1 structure 870:878 a single N-linked oligomannose structure 839:878 a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE 839:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 1 16 theme diseases 143:150 arg1 mediators 121:129 the primary mediators 109:129 the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide 109:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 1 16 theme diseases 143:150 arg1 antibodies 94:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 8 17 theme mast 1160:1163 arg1 degranulation 1170:1182 mast cell degranulation 1160:1182 mast cell degranulation 1160:1182 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 2 18 theme environmental 234:246 arg1 antigens 248:255 innocuous environmental antigens 224:255 innocuous environmental antigens 224:255 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 0 19 theme single 2:7 arg1 glycan 9:14 A single glycan 0:14 A single glycan on IgE 0:21 A single glycan on IgE is indispensable for initiation of anaphylaxis. 25824821 0 19 theme single 2:7 arg1 indispensable 26:38 indispensable 26:38 indispensable 26:38 A single glycan on IgE is indispensable for initiation of anaphylaxis. 25824821 3 20 theme allergic 505:512 arg1 cascade 514:520 the allergic cascade 501:520 the allergic cascade 501:520 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 7 21 theme IgE 914:916 arg1 domain 896:901 the constant domain 3 883:903 the constant domain 3 (Cε3) of IgE 883:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 21 theme IgE 914:916 arg1 Cε3 906:908 Cε3 906:908 Cε3 906:908 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 21 theme IgE 914:916 arg1 IgE 914:916 IgE 914:916 IgE 914:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 1 22 theme Immunoglobulin 71:84 arg1 mediators 121:129 the primary mediators 109:129 the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide 109:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 1 22 theme Immunoglobulin 71:84 arg1 antibodies 94:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 8 23 theme oligomannose 1035:1046 arg1 structure 1077:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 3 24 theme mast 411:414 arg1 IgE 427:429 mast cell-bound IgE 411:429 mast cell-bound IgE 411:429 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 9 25 theme glycosylation 1288:1300 arg1 requirement 1273:1283 an unappreciated and essential requirement 1242:1283 an unappreciated and essential requirement of glycosylation in IgE biology 1242:1315 These results underscore an unappreciated and essential requirement of glycosylation in IgE biology. 25824821 8 26 theme site 1002:1005 arg1 disruption 984:993 Genetic disruption 976:993 Genetic disruption of the site 976:1005 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 8 26 theme site 1002:1005 arg1 removal 1020:1026 enzymatic removal 1010:1026 enzymatic removal of the oligomannose glycan altered IgE secondary structure 1010:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 6 27 theme IgE 762:764 arg1 glycosylation 766:778 IgE glycosylation 762:778 IgE glycosylation 762:778 Here, we demonstrate an absolute requirement for IgE glycosylation in allergic reactions. 25824821 1 28 theme ε 86:86 arg1 mediators 121:129 the primary mediators 109:129 the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide 109:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 1 28 theme ε 86:86 arg1 antibodies 94:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 3 29 theme cell-bound 416:425 arg1 IgE 427:429 mast cell-bound IgE 411:429 mast cell-bound IgE 411:429 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 9 30 theme IgE 1305:1307 arg1 biology 1309:1315 IgE biology 1305:1315 IgE biology 1305:1315 These results underscore an unappreciated and essential requirement of glycosylation in IgE biology. 25824821 9 31 from requirement 1273:1283 arg1 biology 1309:1315 IgE biology 1305:1315 IgE biology 1305:1315 These results underscore an unappreciated and essential requirement of glycosylation in IgE biology. 25824821 4 32 theme precise 551:557 arg1 patterns 573:580 precise glycosylation patterns 551:580 precise glycosylation patterns 551:580 It is well established that precise glycosylation patterns exert profound effects on the biological activity of IgG. 25824821 8 33 theme enzymatic 1010:1018 arg1 removal 1020:1026 enzymatic removal 1010:1026 enzymatic removal of the oligomannose glycan altered IgE secondary structure 1010:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 1 34 theme IgE 89:91 arg1 mediators 121:129 the primary mediators 109:129 the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide 109:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 1 34 theme IgE 89:91 arg1 antibodies 94:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 7 35 theme obligatory 807:816 arg1 glycan 818:823 The obligatory glycan 803:823 The obligatory glycan 803:823 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 1 36 theme primary 113:119 arg1 mediators 121:129 the primary mediators 109:129 the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide 109:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 1 36 theme primary 113:119 arg1 antibodies 94:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies 71:103 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 7 37 from N384 961:964 arg1 mouse 969:973 mouse 969:973 mouse 969:973 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 38 from N394 938:941 arg1 IgE 953:955 human IgE 947:955 human IgE 947:955 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 1 39 from 1 176:176 arg1 worldwide 196:204 10 individuals worldwide 181:204 10 individuals worldwide 181:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 4 40 theme IgG 635:637 arg1 activity 623:630 the biological activity 608:630 the biological activity of IgG 608:637 It is well established that precise glycosylation patterns exert profound effects on the biological activity of IgG. 25824821 3 41 theme Subsequent 370:379 arg1 exposure 390:397 Subsequent allergen exposure 370:397 Subsequent allergen exposure 370:397 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 8 42 theme glycan 1048:1053 arg1 structure 1077:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 8 43 theme secondary 1067:1075 arg1 structure 1077:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 6 44 theme absolute 737:744 arg1 requirement 746:756 an absolute requirement 734:756 an absolute requirement for IgE glycosylation in allergic reactions 734:800 Here, we demonstrate an absolute requirement for IgE glycosylation in allergic reactions. 25824821 3 45 theme mediators 473:481 arg1 initiation 487:496 initiation 487:496 initiation of the allergic cascade 487:520 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 3 45 theme mediators 473:481 arg1 release 449:455 the release 445:455 the release of inflammatory mediators 445:481 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 3 46 theme allergen 381:388 arg1 exposure 390:397 Subsequent allergen exposure 370:397 Subsequent allergen exposure 370:397 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 5 47 theme glycosylation 669:681 arg1 clear 706:710 clear 706:710 clear 706:710 However, the contribution of glycosylation to IgE biology is less clear. 25824821 5 47 theme glycosylation 669:681 arg1 contribution 653:664 the contribution 649:664 the contribution of glycosylation to IgE biology 649:696 However, the contribution of glycosylation to IgE biology is less clear. 25824821 2 48 theme mast 309:312 arg1 cells 314:318 tissue-resident mast cells 293:318 tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI 293:367 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 8 49 theme IgE 1063:1065 arg1 structure 1077:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 6 50 from requirement 746:756 arg1 reactions 792:800 allergic reactions 783:800 allergic reactions 783:800 Here, we demonstrate an absolute requirement for IgE glycosylation in allergic reactions. 25824821 2 51 theme innocuous 224:232 arg1 antigens 248:255 innocuous environmental antigens 224:255 innocuous environmental antigens 224:255 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 8 52 theme altered 1055:1061 arg1 structure 1077:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 the oligomannose glycan altered IgE secondary structure 1031:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 2 53 theme tissue-resident 293:307 arg1 cells 314:318 tissue-resident mast cells 293:318 tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI 293:367 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 5 54 theme IgE 686:688 arg1 biology 690:696 IgE biology 686:696 IgE biology 686:696 However, the contribution of glycosylation to IgE biology is less clear. 25824821 3 55 theme inflammatory 460:471 arg1 mediators 473:481 inflammatory mediators 460:481 inflammatory mediators 460:481 Subsequent allergen exposure cross-links mast cell-bound IgE, resulting in the release of inflammatory mediators and initiation of the allergic cascade. 25824821 7 56 theme constant 887:894 arg1 domain 896:901 the constant domain 3 883:903 the constant domain 3 (Cε3) of IgE 883:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 56 theme constant 887:894 arg1 Cε3 906:908 Cε3 906:908 Cε3 906:908 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 7 56 theme constant 887:894 arg1 IgE 914:916 IgE 914:916 IgE 914:916 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 1 57 theme individuals 184:194 arg1 worldwide 196:204 10 individuals worldwide 181:204 10 individuals worldwide 181:204 Immunoglobulin ε (IgE) antibodies are the primary mediators of allergic diseases, which affect more than 1 in 10 individuals worldwide. 25824821 8 58 theme structure 1077:1085 arg1 disruption 984:993 Genetic disruption 976:993 Genetic disruption of the site 976:1005 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 8 58 theme structure 1077:1085 arg1 removal 1020:1026 enzymatic removal 1010:1026 enzymatic removal of the oligomannose glycan altered IgE secondary structure 1010:1085 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 2 59 theme specific 211:218 arg1 IgE 207:209 IgE 207:209 IgE 207:209 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 7 60 theme human 947:951 arg1 IgE 953:955 human IgE 947:955 human IgE 947:955 The obligatory glycan was mapped to a single N-linked oligomannose structure in the constant domain 3 (Cε3) of IgE, at asparagine-394 (N394) in human IgE and N384 in mouse. 25824821 2 61 theme IgE 349:351 arg1 FcεRI 363:367 FcεRI 363:367 FcεRI 363:367 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 2 61 theme IgE 349:351 arg1 receptor 353:360 the high-affinity IgE receptor 331:360 the high-affinity IgE receptor 331:360 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 4 62 theme biological 612:621 arg1 activity 623:630 the biological activity 608:630 the biological activity of IgG 608:637 It is well established that precise glycosylation patterns exert profound effects on the biological activity of IgG. 25824821 8 63 theme cell 1165:1168 arg1 degranulation 1170:1182 mast cell degranulation 1160:1182 mast cell degranulation 1160:1182 Genetic disruption of the site or enzymatic removal of the oligomannose glycan altered IgE secondary structure and abrogated IgE binding to FcεRI, rendering IgE incapable of eliciting mast cell degranulation, thereby preventing anaphylaxis. 25824821 2 64 theme high-affinity 335:347 arg1 FcεRI 363:367 FcεRI 363:367 FcεRI 363:367 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 25824821 2 64 theme high-affinity 335:347 arg1 receptor 353:360 the high-affinity IgE receptor 331:360 the high-affinity IgE receptor 331:360 IgE specific for innocuous environmental antigens, or allergens, binds and sensitizes tissue-resident mast cells expressing the high-affinity IgE receptor, FcεRI. 24828077 3 0 theme variants 691:698 arg1 range 664:668 an extended range 652:668 an extended range of glycan site viral variants 652:698 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 1 1 theme HIV 250:252 arg1 Env 254:256 HIV Env 250:256 HIV Env 250:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 8 2 theme glycan 1662:1667 arg1 properties 1677:1686 promiscuous glycan binding properties 1650:1686 promiscuous glycan binding properties 1650:1686 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 5 3 contain containing 1164:1173 arg1 viruses 1156:1162 viruses 1156:1162 viruses containing the N332/N334 glycan site 1156:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 3 contain containing 1164:1173 arg2 site 1196:1199 the N332/N334 glycan site 1175:1199 the N332/N334 glycan site 1175:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 4 dep coverage 1215:1222 arg1 % 1213:1213 % 1213:1213 % 1213:1213 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 8 5 theme properties 1677:1686 arg1 importance 1636:1645 the importance 1632:1645 the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities 1632:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 5 6 dep 66 1211:1212 arg1 to 1208:1209 to 1208:1209 to 1208:1209 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 3 7 theme N-linked 745:752 arg1 glycans 754:760 alternate N-linked glycans 735:760 alternate N-linked glycans 735:760 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 1 8 theme high-mannose 185:196 arg1 patch 198:202 the high-mannose patch 181:202 the high-mannose patch centered around the glycan at position 332 on HIV Env 181:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 4 9 theme antibodies 901:910 arg1 many 889:892 many 889:892 many 889:892 Furthermore, many of the antibodies can neutralize viruses in which the N332 glycan site is shifted to the 334 position. 24828077 4 9 theme antibodies 901:910 arg1 antibodies 901:910 the antibodies 897:910 the antibodies 897:910 Furthermore, many of the antibodies can neutralize viruses in which the N332 glycan site is shifted to the 334 position. 24828077 7 10 theme glycosylation 1581:1593 arg1 sites 1595:1599 glycosylation sites 1581:1599 glycosylation sites 1581:1599 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 2 11 from degrees 502:508 arg1 site 543:546 the N332 glycan site 527:546 the N332 glycan site 527:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 8 12 from activity 1746:1753 arg1 modalities 1791:1800 either protective or therapeutic modalities 1758:1800 either protective or therapeutic modalities 1758:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 2 13 theme breadth 589:595 arg1 obvious 612:618 obvious 612:618 obvious 612:618 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 2 13 theme breadth 589:595 arg1 origins 557:563 the origins 553:563 the origins of their neutralization breadth 553:595 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 2 14 from dependency 513:522 arg1 site 543:546 the N332 glycan site 527:546 the N332 glycan site 527:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 1 15 theme established 415:425 arg1 infection 427:435 established infection 415:435 established infection in macaque models 415:453 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 8 16 theme optimal 1728:1734 arg1 activity 1746:1753 optimal antiviral activity 1728:1753 optimal antiviral activity in either protective or therapeutic modalities 1728:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 3 17 theme glycan 789:794 arg1 site 796:799 the N332 glycan site 780:799 the N332 glycan site 780:799 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 1 18 theme monoclonal 138:147 arg1 bnmAbs 161:166 bnmAbs 161:166 bnmAbs 161:166 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 18 theme monoclonal 138:147 arg1 vaccine 272:278 promising vaccine leads 262:284 promising vaccine leads 262:284 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 18 theme monoclonal 138:147 arg1 antibodies 149:158 Broadly neutralizing monoclonal antibodies 117:158 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env 117:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 3 19 theme substantial 828:838 arg1 number 840:845 a substantial number 826:845 a substantial number of viruses lacking the site 826:873 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 7 20 theme neutralization 1535:1548 arg1 escape 1550:1555 neutralization escape 1535:1555 neutralization escape mediated by shifting of glycosylation sites 1535:1599 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 4 21 theme glycan 953:958 arg1 site 960:963 the N332 glycan site 944:963 the N332 glycan site 944:963 Furthermore, many of the antibodies can neutralize viruses in which the N332 glycan site is shifted to the 334 position. 24828077 0 22 theme high-mannose 57:68 arg1 patch 70:74 the high-mannose patch 53:74 the high-mannose patch of gp120 53:83 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 5 23 theme high-mannose 1077:1088 arg1 patch 1090:1094 the high-mannose patch 1073:1094 the high-mannose patch 1073:1094 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 6 24 theme near-equivalent 1361:1375 arg1 coverage 1377:1384 near-equivalent coverage 1361:1384 near-equivalent coverage 1361:1384 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 6 24 theme near-equivalent 1361:1375 arg1 combination 1391:1401 a combination 1389:1401 a combination of bnmAbs targeting multiple epitopes 1389:1439 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 1 25 from infection 427:435 arg1 models 448:453 macaque models 440:453 macaque models 440:453 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 26 dep vaccine 272:278 arg1 leads 280:284 leads 280:284 leads 280:284 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 7 27 link N-linked 1496:1503 arg1 sites 1512:1516 other N-linked glycan sites 1490:1516 other N-linked glycan sites 1490:1516 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 5 28 theme families 1052:1059 arg1 combination 1022:1032 a combination 1020:1032 a combination of three antibody families that target the high-mannose patch 1020:1094 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 7 29 theme N-linked 1496:1503 arg1 sites 1512:1516 other N-linked glycan sites 1490:1516 other N-linked glycan sites 1490:1516 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 2 30 theme N332 531:534 arg1 site 543:546 the N332 glycan site 527:546 the N332 glycan site 527:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 0 31 theme Promiscuous 0:10 arg1 recognition 24:34 Promiscuous glycan site recognition 0:34 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 0:83 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 1 32 theme promising 262:270 arg1 vaccine 272:278 promising vaccine leads 262:284 promising vaccine leads 262:284 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 32 theme promising 262:270 arg1 antibodies 149:158 Broadly neutralizing monoclonal antibodies 117:158 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env 117:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 0 33 theme site 19:22 arg1 recognition 24:34 Promiscuous glycan site recognition 0:34 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 0:83 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 2 34 theme varying 494:500 arg1 degrees 502:508 varying degrees 494:508 varying degrees of dependency on the N332 glycan site 494:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 5 35 theme N332/N334 1250:1258 arg1 site 1267:1270 the N332/N334 glycan site 1246:1270 the N332/N334 glycan site 1246:1270 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 1 36 theme therapeutic 290:300 arg1 candidates 302:311 therapeutic candidates 290:311 therapeutic candidates 290:311 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 4 37 theme N332 948:951 arg1 site 960:963 the N332 glycan site 944:963 the N332 glycan site 944:963 Furthermore, many of the antibodies can neutralize viruses in which the N332 glycan site is shifted to the 334 position. 24828077 6 38 theme high-mannose 1330:1341 arg1 patch 1343:1347 the high-mannose patch 1326:1347 the high-mannose patch 1326:1347 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 5 39 theme % 1110:1110 arg1 coverage 1127:1134 99% neutralization coverage 1108:1134 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site 1108:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 3 40 theme viral 685:689 arg1 variants 691:698 glycan site viral variants 673:698 glycan site viral variants 673:698 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 3 41 theme glycan 673:678 arg1 variants 691:698 glycan site viral variants 673:698 glycan site viral variants 673:698 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 6 42 theme multiple 1423:1430 arg1 epitopes 1432:1439 multiple epitopes 1423:1439 multiple epitopes 1423:1439 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 5 43 theme glycan 1189:1194 arg1 site 1196:1199 the N332/N334 glycan site 1175:1199 the N332/N334 glycan site 1175:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 7 44 gly glycosylation 1581:1593 arg2 sites 1595:1599 glycosylation sites 1581:1599 glycosylation sites 1581:1599 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 6 45 theme bnmAbs 1406:1411 arg1 coverage 1377:1384 near-equivalent coverage 1361:1384 near-equivalent coverage 1361:1384 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 6 45 theme bnmAbs 1406:1411 arg1 combination 1391:1401 a combination 1389:1401 a combination of bnmAbs targeting multiple epitopes 1389:1439 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 5 46 theme panel 1147:1151 arg1 coverage 1215:1222 up to 66% coverage 1205:1222 up to 66% coverage for viruses that lack the N332/N334 glycan site 1205:1270 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 46 theme panel 1147:1151 arg1 coverage 1127:1134 99% neutralization coverage 1108:1134 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site 1108:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 47 theme viruses 1156:1162 arg1 panel 1147:1151 a large panel 1139:1151 a large panel of viruses containing the N332/N334 glycan site 1139:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 48 dep % 1213:1213 arg1 66 1211:1212 66 1211:1212 66 1211:1212 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 8 49 theme promiscuous 1650:1660 arg1 properties 1677:1686 promiscuous glycan binding properties 1650:1686 promiscuous glycan binding properties 1650:1686 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 6 50 theme diverse 1301:1307 arg1 response 1309:1316 a diverse response 1299:1316 a diverse response against the high-mannose patch 1299:1347 The results indicate that a diverse response against the high-mannose patch may provide near-equivalent coverage as a combination of bnmAbs targeting multiple epitopes. 24828077 8 51 theme binding 1669:1675 arg1 properties 1677:1686 promiscuous glycan binding properties 1650:1686 promiscuous glycan binding properties 1650:1686 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 7 52 theme sites 1595:1599 arg1 shifting 1569:1576 shifting 1569:1576 shifting of glycosylation sites 1569:1599 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 3 53 theme alternate 735:743 arg1 glycans 754:760 alternate N-linked glycans 735:760 alternate N-linked glycans 735:760 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 1 54 theme SHIV 399:402 arg1 viremia 404:410 SHIV viremia 399:410 SHIV viremia 399:410 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 55 from position 234:241 arg1 glycan 224:229 the glycan 220:229 the glycan at position 332 on HIV Env 220:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 5 56 theme antibody 1043:1050 arg1 families 1052:1059 three antibody families 1037:1059 three antibody families that target the high-mannose patch 1037:1094 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 8 57 theme high-mannose 1705:1716 arg1 patch 1718:1722 the high-mannose patch 1701:1722 the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities 1701:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 2 58 theme neutralization 574:587 arg1 breadth 589:595 their neutralization breadth 568:595 their neutralization breadth 568:595 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 1 59 from glycan 224:229 arg1 Env 254:256 HIV Env 250:256 HIV Env 250:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 60 theme neutralizing 125:136 arg1 bnmAbs 161:166 bnmAbs 161:166 bnmAbs 161:166 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 60 theme neutralizing 125:136 arg1 vaccine 272:278 promising vaccine leads 262:284 promising vaccine leads 262:284 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 1 60 theme neutralizing 125:136 arg1 antibodies 149:158 Broadly neutralizing monoclonal antibodies 117:158 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env 117:256 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 3 61 theme viruses 850:856 arg1 number 840:845 a substantial number 826:845 a substantial number of viruses lacking the site 826:873 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 2 62 from site 543:546 arg1 degrees 502:508 varying degrees 494:508 varying degrees of dependency on the N332 glycan site 494:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 3 63 theme site 796:799 arg1 absence 769:775 the absence 765:775 the absence of the N332 glycan site 765:799 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 1 64 theme macaque 440:446 arg1 models 448:453 macaque models 440:453 macaque models 440:453 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 8 65 theme antiviral 1736:1744 arg1 activity 1746:1753 optimal antiviral activity 1728:1753 optimal antiviral activity in either protective or therapeutic modalities 1728:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 2 66 theme dependency 513:522 arg1 degrees 502:508 varying degrees 494:508 varying degrees of dependency on the N332 glycan site 494:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 3 67 link N-linked 745:752 arg1 glycans 754:760 alternate N-linked glycans 735:760 alternate N-linked glycans 735:760 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 3 68 theme N332 784:787 arg1 site 796:799 the N332 glycan site 780:799 the N332 glycan site 780:799 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 0 69 theme gp120 79:83 arg1 patch 70:74 the high-mannose patch 53:74 the high-mannose patch of gp120 53:83 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 1 70 theme SHIV 362:365 arg1 challenge 367:375 mucosal SHIV challenge 354:375 mucosal SHIV challenge 354:375 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 7 71 theme glycan 1505:1510 arg1 sites 1512:1516 other N-linked glycan sites 1490:1516 other N-linked glycan sites 1490:1516 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 1 72 theme mucosal 354:360 arg1 challenge 367:375 mucosal SHIV challenge 354:375 mucosal SHIV challenge 354:375 Broadly neutralizing monoclonal antibodies (bnmAbs) that target the high-mannose patch centered around the glycan at position 332 on HIV Env are promising vaccine leads and therapeutic candidates because they effectively protect against mucosal SHIV challenge and strongly suppress SHIV viremia in established infection in macaque models. 24828077 8 73 theme protective 1765:1774 arg1 modalities 1791:1800 either protective or therapeutic modalities 1758:1800 either protective or therapeutic modalities 1758:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 7 74 theme other 1490:1494 arg1 sites 1512:1516 other N-linked glycan sites 1490:1516 other N-linked glycan sites 1490:1516 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 2 75 theme glycan 536:541 arg1 site 543:546 the N332 glycan site 527:546 the N332 glycan site 527:546 However, these antibodies demonstrate varying degrees of dependency on the N332 glycan site, and the origins of their neutralization breadth are not always obvious. 24828077 5 76 theme glycan 1260:1265 arg1 site 1267:1270 the N332/N334 glycan site 1246:1270 the N332/N334 glycan site 1246:1270 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 0 77 theme HIV 112:114 arg1 neutralization 94:107 neutralization 94:107 neutralization of HIV 94:114 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 8 78 theme therapeutic 1779:1789 arg1 modalities 1791:1800 either protective or therapeutic modalities 1758:1800 either protective or therapeutic modalities 1758:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 0 79 theme glycan 12:17 arg1 recognition 24:34 Promiscuous glycan site recognition 0:34 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 0:83 Promiscuous glycan site recognition by antibodies to the high-mannose patch of gp120 broadens neutralization of HIV. 24828077 8 80 from importance 1636:1645 arg1 bnmAbs 1691:1696 bnmAbs 1691:1696 bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities 1691:1800 Overall, this work highlights the importance of promiscuous glycan binding properties in bnmAbs to the high-mannose patch for optimal antiviral activity in either protective or therapeutic modalities. 24828077 3 81 theme extended 655:662 arg1 range 664:668 an extended range 652:668 an extended range of glycan site viral variants 652:698 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 5 82 theme large 1141:1145 arg1 panel 1147:1151 a large panel 1139:1151 a large panel of viruses containing the N332/N334 glycan site 1139:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 83 theme 99 1108:1109 arg1 % 1110:1110 % 1110:1110 % 1110:1110 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 7 84 theme bnmAbs 1476:1481 arg1 ability 1460:1466 the ability 1456:1466 the ability of some bnmAbs to use other N-linked glycan sites 1456:1516 Additionally, the ability of some bnmAbs to use other N-linked glycan sites can help counter neutralization escape mediated by shifting of glycosylation sites. 24828077 3 85 theme site 680:683 arg1 variants 691:698 glycan site viral variants 673:698 glycan site viral variants 673:698 By measuring neutralization on an extended range of glycan site viral variants, we found that some bnmAbs can use alternate N-linked glycans in the absence of the N332 glycan site and therefore neutralize a substantial number of viruses lacking the site. 24828077 5 86 theme neutralization 1112:1125 arg1 coverage 1127:1134 99% neutralization coverage 1108:1134 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site 1108:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24828077 5 87 theme N332/N334 1179:1187 arg1 site 1196:1199 the N332/N334 glycan site 1175:1199 the N332/N334 glycan site 1175:1199 Finally, we found that a combination of three antibody families that target the high-mannose patch can lead to 99% neutralization coverage of a large panel of viruses containing the N332/N334 glycan site and up to 66% coverage for viruses that lack the N332/N334 glycan site. 24497634 5 0 theme molecular 755:763 arg1 determinants 765:776 the molecular determinants 751:776 the molecular determinants of contactin targeted by the autoantibodies 751:820 In the present study, we mapped the molecular determinants of contactin targeted by the autoantibodies. 24497634 6 1 theme contactin 898:906 arg1 contactin 898:906 contactin 898:906 contactin 898:906 In three patients, immunoreactivity was directed against the Ig domains of contactin and was dependent on N-glycans. 24497634 6 1 theme contactin 898:906 arg1 domains 887:893 the Ig domains 880:893 the Ig domains of contactin 880:906 In three patients, immunoreactivity was directed against the Ig domains of contactin and was dependent on N-glycans. 24497634 1 2 theme rapid 223:227 arg1 propagation 229:239 the rapid propagation 219:239 the rapid propagation of the nerve impulses along myelinated axons 219:284 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 15 3 theme blocking 2032:2039 arg1 activity 2041:2048 functional blocking activity 2021:2048 functional blocking activity 2021:2048 These results strongly suggest that antibodies to CAMs may be pathogenic and induce demyelination via functional blocking activity. 24497634 14 4 theme junctions 1877:1885 arg1 alteration 1853:1862 alteration 1853:1862 alteration of paranodal junctions 1853:1885 Importantly, we showed that the anti-contactin autoantibodies induced alteration of paranodal junctions in myelinated neuronal culture. 24497634 12 5 dep sites 1546:1550 arg1 sites 1546:1550 N-glycosylation sites 1530:1550 N-glycosylation sites (Asn-467, Asn-473, and Asn-494) 1530:1582 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 5 dep sites 1546:1550 arg1 Asn-473 1562:1568 Asn-473 1562:1568 Asn-473 1562:1568 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 5 dep sites 1546:1550 arg1 Asn-494 1575:1581 Asn-494 1575:1581 Asn-494 1575:1581 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 5 dep sites 1546:1550 arg1 Asn-467 1553:1559 Asn-467 1553:1559 Asn-467 1553:1559 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 7 6 theme patient 957:963 arg1 serum 944:948 The serum 940:948 The serum of one patient 940:963 The serum of one patient was selectively directed against contactin bearing mannose-rich N-glycans. 24497634 1 7 theme crucial 158:164 arg1 role 166:169 a crucial role 156:169 a crucial role 156:169 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 5 8 theme present 726:732 arg1 study 734:738 the present study 722:738 the present study 722:738 In the present study, we mapped the molecular determinants of contactin targeted by the autoantibodies. 24497634 2 9 theme autoimmunity 317:328 arg1 targets 306:312 the targets 302:312 the targets of autoimmunity in inflammatory neuropathies 302:357 These CAMs are the targets of autoimmunity in inflammatory neuropathies. 24497634 2 9 theme autoimmunity 317:328 arg1 CAMs 293:296 These CAMs 287:296 These CAMs 287:296 These CAMs are the targets of autoimmunity in inflammatory neuropathies. 24497634 10 10 theme contactin 1375:1383 arg1 contactin 1375:1383 contactin 1375:1383 contactin 1375:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 10 10 theme contactin 1375:1383 arg1 domain 1363:1368 Ig domain 5 1360:1370 Ig domain 5 of contactin 1360:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 8 11 theme oligomannose 1056:1067 arg1 sugars 1074:1079 the oligomannose type sugars 1052:1079 the oligomannose type sugars of contactin 1052:1092 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 0 12 theme peripheral 95:104 arg1 neuropathies 106:117 peripheral neuropathies 95:117 peripheral neuropathies 95:117 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 0 13 from binding 59:65 arg1 neuropathies 106:117 peripheral neuropathies 95:117 peripheral neuropathies 95:117 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 13 14 theme aggregation 1645:1655 arg1 assays 1657:1662 cell aggregation assays 1640:1662 cell aggregation assays 1640:1662 Using cell aggregation assays, we showed that the IgGs from the four CIDP patients prevented adhesive interaction between contactin·Caspr and NF155. 24497634 2 15 from targets 306:312 arg1 neuropathies 346:357 inflammatory neuropathies 333:357 inflammatory neuropathies 333:357 These CAMs are the targets of autoimmunity in inflammatory neuropathies. 24497634 3 16 dep shows 488:492 arg1 1 523:523 1 523:523 1 523:523 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 17 theme aggressive 412:421 arg1 CIDP 482:485 CIDP 482:485 CIDP 482:485 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 17 theme aggressive 412:421 arg1 polyradiculoneuropathy 458:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy 412:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 412:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 13 18 theme CIDP 1703:1706 arg1 patients 1708:1715 the four CIDP patients 1694:1715 the four CIDP patients 1694:1715 Using cell aggregation assays, we showed that the IgGs from the four CIDP patients prevented adhesive interaction between contactin·Caspr and NF155. 24497634 13 19 from patients 1708:1715 arg1 IgGs 1684:1687 the IgGs 1680:1687 the IgGs from the four CIDP patients 1680:1715 Using cell aggregation assays, we showed that the IgGs from the four CIDP patients prevented adhesive interaction between contactin·Caspr and NF155. 24497634 8 20 theme type 1069:1072 arg1 sugars 1074:1079 the oligomannose type sugars 1052:1079 the oligomannose type sugars of contactin 1052:1092 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 9 21 theme N-glycans 1204:1212 arg1 role 1186:1189 the role 1182:1189 the role of contactin N-glycans 1182:1212 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 3 22 theme chronic 423:429 arg1 CIDP 482:485 CIDP 482:485 CIDP 482:485 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 22 theme chronic 423:429 arg1 polyradiculoneuropathy 458:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy 412:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 412:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 0 23 theme contactin 9:17 arg1 N-glycans 19:27 Specific contactin N-glycans 0:27 Specific contactin N-glycans 0:27 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 6 24 theme Ig 884:885 arg1 contactin 898:906 contactin 898:906 contactin 898:906 In three patients, immunoreactivity was directed against the Ig domains of contactin and was dependent on N-glycans. 24497634 6 24 theme Ig 884:885 arg1 domains 887:893 the Ig domains 880:893 the Ig domains of contactin 880:906 In three patients, immunoreactivity was directed against the Ig domains of contactin and was dependent on N-glycans. 24497634 8 25 theme contactin 1084:1092 arg1 sugars 1074:1079 the oligomannose type sugars 1052:1079 the oligomannose type sugars of contactin 1052:1092 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 4 26 theme contactin·Caspr·neurofascin-155 542:572 arg1 complex 531:537 The complex 527:537 The complex of contactin·Caspr·neurofascin-155 (NF155) 527:580 The complex of contactin·Caspr·neurofascin-155 (NF155) enables the formation of paranodal junctions, suggesting that antibody attack against paranodes may participate in the severity of CIDP. 24497634 3 27 theme patients 398:405 arg1 subgroup 386:393 a subgroup 384:393 a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 384:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 0 28 theme Specific 0:7 arg1 N-glycans 19:27 Specific contactin N-glycans 0:27 Specific contactin N-glycans 0:27 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 1 29 theme nerve 248:252 arg1 impulses 254:261 the nerve impulses 244:261 the nerve impulses along myelinated axons 244:284 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 10 30 theme NF155-Fc 1403:1410 arg1 binding 1412:1418 soluble NF155-Fc binding 1395:1418 soluble NF155-Fc binding 1395:1418 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 14 31 theme neuronal 1901:1908 arg1 culture 1910:1916 myelinated neuronal culture 1890:1916 myelinated neuronal culture 1890:1916 Importantly, we showed that the anti-contactin autoantibodies induced alteration of paranodal junctions in myelinated neuronal culture. 24497634 13 32 theme adhesive 1727:1734 arg1 interaction 1736:1746 adhesive interaction 1727:1746 adhesive interaction between contactin·Caspr and NF155 1727:1780 Using cell aggregation assays, we showed that the IgGs from the four CIDP patients prevented adhesive interaction between contactin·Caspr and NF155. 24497634 1 33 theme impulses 254:261 arg1 propagation 229:239 the rapid propagation 219:239 the rapid propagation of the nerve impulses along myelinated axons 219:284 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 9 34 gly N-glycosylation 1258:1272 arg2 sites 1274:1278 the nine consensus N-glycosylation sites 1239:1278 the nine consensus N-glycosylation sites 1239:1278 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 9 34 gly N-glycosylation 1258:1272 arg2 nine 1243:1246 nine 1243:1246 nine 1243:1246 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 9 35 theme contactin 1194:1202 arg1 N-glycans 1204:1212 contactin N-glycans 1194:1212 contactin N-glycans 1194:1212 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 12 36 from immunoreactivity 1601:1616 arg1 patient 1625:1631 one patient 1621:1631 one patient 1621:1631 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 37 theme sites 1546:1550 arg1 cluster 1519:1525 the cluster 1515:1525 the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) 1515:1582 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 14 38 theme myelinated 1890:1899 arg1 culture 1910:1916 myelinated neuronal culture 1890:1916 myelinated neuronal culture 1890:1916 Importantly, we showed that the anti-contactin autoantibodies induced alteration of paranodal junctions in myelinated neuronal culture. 24497634 4 39 theme paranodal 607:615 arg1 junctions 617:625 paranodal junctions 607:625 paranodal junctions 607:625 The complex of contactin·Caspr·neurofascin-155 (NF155) enables the formation of paranodal junctions, suggesting that antibody attack against paranodes may participate in the severity of CIDP. 24497634 11 40 with cis-association 1466:1480 arg1 Caspr 1487:1491 Caspr 1487:1491 Caspr 1487:1491 In contrast, these mutations did not abolish cis-association with Caspr. 24497634 1 41 theme Cell 120:123 arg1 CAMs 145:148 CAMs 145:148 CAMs 145:148 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 1 41 theme Cell 120:123 arg1 molecules 134:142 Cell adhesion molecules 120:142 Cell adhesion molecules (CAMs) 120:149 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 1 42 theme myelinated 269:278 arg1 axons 280:284 myelinated axons 269:284 myelinated axons 269:284 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 0 43 from targeting 82:90 arg1 neuropathies 106:117 peripheral neuropathies 95:117 peripheral neuropathies 95:117 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 12 44 gly N-glycosylation 1530:1544 arg2 Asn-467 1553:1559 Asn-467 1553:1559 Asn-467 1553:1559 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 44 gly N-glycosylation 1530:1544 arg2 sites 1546:1550 N-glycosylation sites 1530:1550 N-glycosylation sites (Asn-467, Asn-473, and Asn-494) 1530:1582 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 44 gly N-glycosylation 1530:1544 arg2 Asn-494 1575:1581 Asn-494 1575:1581 Asn-494 1575:1581 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 44 gly N-glycosylation 1530:1544 arg2 Asn-473 1562:1568 Asn-473 1562:1568 Asn-473 1562:1568 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 1 45 theme nodes 195:199 arg1 formation 178:186 the formation 174:186 the formation of the nodes of Ranvier 174:210 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 5 46 theme contactin 781:789 arg1 determinants 765:776 the molecular determinants 751:776 the molecular determinants of contactin targeted by the autoantibodies 751:820 In the present study, we mapped the molecular determinants of contactin targeted by the autoantibodies. 24497634 13 47 theme cell 1640:1643 arg1 assays 1657:1662 cell aggregation assays 1640:1662 cell aggregation assays 1640:1662 Using cell aggregation assays, we showed that the IgGs from the four CIDP patients prevented adhesive interaction between contactin·Caspr and NF155. 24497634 3 48 theme demyelinating 444:456 arg1 CIDP 482:485 CIDP 482:485 CIDP 482:485 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 48 theme demyelinating 444:456 arg1 polyradiculoneuropathy 458:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy 412:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 412:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 49 with patients 398:405 arg1 CIDP 482:485 CIDP 482:485 CIDP 482:485 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 49 with patients 398:405 arg1 polyradiculoneuropathy 458:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy 412:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 412:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 1 50 theme adhesion 125:132 arg1 CAMs 145:148 CAMs 145:148 CAMs 145:148 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 1 50 theme adhesion 125:132 arg1 molecules 134:142 Cell adhesion molecules 120:142 Cell adhesion molecules (CAMs) 120:149 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 0 51 theme neurofascin 47:57 arg1 binding 59:65 neurofascin binding 47:65 neurofascin binding 47:65 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 15 52 theme functional 2021:2030 arg1 activity 2041:2048 functional blocking activity 2021:2048 functional blocking activity 2021:2048 These results strongly suggest that antibodies to CAMs may be pathogenic and induce demyelination via functional blocking activity. 24497634 10 53 theme sites 1331:1335 arg1 mutation 1313:1320 the mutation 1309:1320 the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin 1309:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 4 54 theme CIDP 713:716 arg1 severity 701:708 the severity 697:708 the severity of CIDP 697:716 The complex of contactin·Caspr·neurofascin-155 (NF155) enables the formation of paranodal junctions, suggesting that antibody attack against paranodes may participate in the severity of CIDP. 24497634 8 55 with association 1111:1121 arg1 2 1153:1153 2 1153:1153 2 1153:1153 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 8 55 with association 1111:1121 arg1 NF155 1146:1150 its glial partner NF155 1128:1150 its glial partner NF155 (2) 1128:1154 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 10 56 theme soluble 1395:1401 arg1 binding 1412:1418 soluble NF155-Fc binding 1395:1418 soluble NF155-Fc binding 1395:1418 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 8 57 theme partner 1138:1144 arg1 2 1153:1153 2 1153:1153 2 1153:1153 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 8 57 theme partner 1138:1144 arg1 NF155 1146:1150 its glial partner NF155 1128:1150 its glial partner NF155 (2) 1128:1154 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 9 58 theme consensus 1248:1256 arg1 sites 1274:1278 the nine consensus N-glycosylation sites 1239:1278 the nine consensus N-glycosylation sites 1239:1278 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 7 59 theme mannose-rich 1016:1027 arg1 N-glycans 1029:1037 mannose-rich N-glycans 1016:1037 mannose-rich N-glycans 1016:1037 The serum of one patient was selectively directed against contactin bearing mannose-rich N-glycans. 24497634 1 60 theme Ranvier 204:210 arg1 nodes 195:199 the nodes 191:199 the nodes of Ranvier 191:210 Cell adhesion molecules (CAMs) play a crucial role in the formation of the nodes of Ranvier and in the rapid propagation of the nerve impulses along myelinated axons. 24497634 8 61 theme glial 1132:1136 arg1 2 1153:1153 2 1153:1153 2 1153:1153 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 8 61 theme glial 1132:1136 arg1 NF155 1146:1150 its glial partner NF155 1128:1150 its glial partner NF155 (2) 1128:1154 Strikingly, the oligomannose type sugars of contactin are required for association with its glial partner NF155 (2). 24497634 9 62 theme N-glycosylation 1258:1272 arg1 sites 1274:1278 the nine consensus N-glycosylation sites 1239:1278 the nine consensus N-glycosylation sites 1239:1278 To investigate precisely the role of contactin N-glycans, we have mutated each of the nine consensus N-glycosylation sites independently. 24497634 14 63 theme paranodal 1867:1875 arg1 junctions 1877:1885 paranodal junctions 1867:1885 paranodal junctions 1867:1885 Importantly, we showed that the anti-contactin autoantibodies induced alteration of paranodal junctions in myelinated neuronal culture. 24497634 12 64 theme N-glycosylation 1530:1544 arg1 sites 1546:1550 N-glycosylation sites 1530:1550 N-glycosylation sites (Asn-467, Asn-473, and Asn-494) 1530:1582 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 64 theme N-glycosylation 1530:1544 arg1 Asn-473 1562:1568 Asn-473 1562:1568 Asn-473 1562:1568 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 64 theme N-glycosylation 1530:1544 arg1 Asn-494 1575:1581 Asn-494 1575:1581 Asn-494 1575:1581 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 12 64 theme N-glycosylation 1530:1544 arg1 Asn-467 1553:1559 Asn-467 1553:1559 Asn-467 1553:1559 Next, we showed that the cluster of N-glycosylation sites (Asn-467, Asn-473, and Asn-494) was required for immunoreactivity in one patient. 24497634 0 65 theme autoimmune 71:80 arg1 targeting 82:90 autoimmune targeting 71:90 autoimmune targeting 71:90 Specific contactin N-glycans are implicated in neurofascin binding and autoimmune targeting in peripheral neuropathies. 24497634 4 66 theme antibody 644:651 arg1 attack 653:658 antibody attack 644:658 antibody attack against paranodes 644:676 The complex of contactin·Caspr·neurofascin-155 (NF155) enables the formation of paranodal junctions, suggesting that antibody attack against paranodes may participate in the severity of CIDP. 24497634 3 67 theme inflammatory 431:442 arg1 CIDP 482:485 CIDP 482:485 CIDP 482:485 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 3 67 theme inflammatory 431:442 arg1 polyradiculoneuropathy 458:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy 412:479 aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) 412:486 We recently showed that a subgroup of patients with aggressive chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) shows autoantibodies to contactin (1). 24497634 10 68 theme Ig 1360:1361 arg1 contactin 1375:1383 contactin 1375:1383 contactin 1375:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 10 68 theme Ig 1360:1361 arg1 domain 1363:1368 Ig domain 5 1360:1370 Ig domain 5 of contactin 1360:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 14 69 theme anti-contactin 1815:1828 arg1 autoantibodies 1830:1843 the anti-contactin autoantibodies 1811:1843 the anti-contactin autoantibodies 1811:1843 Importantly, we showed that the anti-contactin autoantibodies induced alteration of paranodal junctions in myelinated neuronal culture. 24497634 10 70 from mutation 1313:1320 arg1 contactin 1375:1383 contactin 1375:1383 contactin 1375:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 10 70 from mutation 1313:1320 arg1 domain 1363:1368 Ig domain 5 1360:1370 Ig domain 5 of contactin 1360:1383 We found that the mutation of three sites (N467Q/N473Q/N494Q) in Ig domain 5 of contactin prevented soluble NF155-Fc binding. 24497634 2 71 theme inflammatory 333:344 arg1 neuropathies 346:357 inflammatory neuropathies 333:357 inflammatory neuropathies 333:357 These CAMs are the targets of autoimmunity in inflammatory neuropathies. 24497634 4 72 theme junctions 617:625 arg1 formation 594:602 the formation 590:602 the formation of paranodal junctions 590:625 The complex of contactin·Caspr·neurofascin-155 (NF155) enables the formation of paranodal junctions, suggesting that antibody attack against paranodes may participate in the severity of CIDP. 25253346 13 0 theme Fine 1822:1825 arg1 mapping 1827:1833 Fine mapping 1822:1833 Fine mapping at the CD4bs 1822:1846 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 11 1 theme limited 1628:1634 arg1 activity 1651:1658 limited neutralization activity 1628:1658 limited neutralization activity 1628:1658 However, the core-elicited sera showed limited neutralization activity. 25253346 19 2 theme glycan 2972:2977 arg1 masking 2979:2985 glycan masking 2972:2985 glycan masking 2972:2985 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 6 3 gly glycoproteins 836:848 arg1 glycoproteins 836:848 most glycoproteins 831:848 most glycoproteins 831:848 Following purification, most glycoproteins, except for 10G, were recognized by broadly neutralizing CD4bs-directed antibodies. 25253346 2 4 theme primary 314:320 arg1 CD4bs 349:353 CD4bs 349:353 CD4bs 349:353 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 4 theme primary 314:320 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 4 theme primary 314:320 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 18 5 theme atomic-level 2800:2811 arg1 structure 2813:2821 the atomic-level structure 2796:2821 the atomic-level structure 2796:2821 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 4 6 theme cysteine-stabilized 565:583 arg1 core 591:594 a cysteine-stabilized gp120 core 563:594 a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs 563:695 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 4 6 theme cysteine-stabilized 565:583 arg1 0G 597:598 0G 597:598 0G 597:598 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 10 7 theme CD4bs-directed 1553:1566 arg1 specificity 1576:1586 CD4bs-directed binding specificity 1553:1586 CD4bs-directed binding specificity 1553:1586 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 1 8 theme host 236:239 arg1 cells 241:245 CD4(+) CCR5(+) host cells 221:245 CD4(+) CCR5(+) host cells 221:245 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 4 9 theme variable 622:629 arg1 regions 631:637 its major variable regions 612:637 its major variable regions to preferentially expose the conformationally fixed CD4bs 612:695 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 9 10 theme parental 1428:1435 arg1 0G 1437:1438 parental 0G 1428:1438 parental 0G 1428:1438 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 8 11 theme neutralizing 1128:1139 arg1 antibodies 1156:1165 seven broadly neutralizing CD4bs-directed antibodies 1114:1165 seven broadly neutralizing CD4bs-directed antibodies 1114:1165 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 14 12 theme envelope 2230:2237 arg1 glycoproteins 2247:2259 envelope surface glycoproteins 2230:2259 envelope surface glycoproteins 2230:2259 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 14 13 dep glycan 2282:2287 arg1 shield 2289:2294 shield 2289:2294 shield 2289:2294 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 8 14 theme neutralizing 1204:1215 arg1 antibodies 1232:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 19 15 theme unmodified 3125:3134 arg1 gp120 3136:3140 unmodified gp120 3125:3140 unmodified gp120 3125:3140 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 16 16 theme binding 2593:2599 arg1 site 2601:2604 this gp120 CD4 binding site 2578:2604 this gp120 CD4 binding site 2578:2604 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 13 17 theme Env 1964:1966 arg1 sites 1968:1972 Env sites 1964:1972 Env sites of vulnerability to neutralizing antibody 1964:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 13 17 theme Env 1964:1966 arg1 vulnerability 1977:1989 vulnerability 1977:1989 vulnerability to neutralizing antibody 1977:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 14 18 theme effective 2175:2183 arg1 vaccine 2191:2197 an effective HIV-1 vaccine 2172:2197 developing an effective HIV-1 vaccine 2161:2197 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 7 19 theme additional 982:991 arg1 N-glycans 993:1001 additional N-glycans 982:1001 additional N-glycans 982:1001 Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. 25253346 2 20 theme conserved 262:270 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 20 theme conserved 262:270 arg1 determinants 282:293 conserved antigenic determinants 262:293 conserved antigenic determinants 262:293 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 16 21 dep target 2571:2576 arg1 CD4bs 2607:2611 CD4bs 2607:2611 CD4bs 2607:2611 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 3 22 contain contains 397:404 arg1 Env 388:390 Env 388:390 Env 388:390 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 3 22 contain contains 397:404 arg2 regions 415:421 variable regions 406:421 variable regions 406:421 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 3 22 contain contains 397:404 arg2 surfaces 435:442 protein surfaces 427:442 protein surfaces 427:442 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 14 23 theme developing 2161:2170 arg1 vaccine 2191:2197 an effective HIV-1 vaccine 2172:2197 developing an effective HIV-1 vaccine 2161:2197 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 15 24 theme sites 2390:2394 arg1 one 2446:2448 one 2446:2448 one 2446:2448 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 24 theme sites 2390:2394 arg1 site 2495:2498 the virus receptor (CD4) binding site 2462:2498 the virus receptor (CD4) binding site 2462:2498 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 24 theme sites 2390:2394 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 25 theme CD4 2482:2484 arg1 one 2446:2448 one 2446:2448 one 2446:2448 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 25 theme CD4 2482:2484 arg1 site 2495:2498 the virus receptor (CD4) binding site 2462:2498 the virus receptor (CD4) binding site 2462:2498 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 25 theme CD4 2482:2484 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 1 26 theme CD4 221:223 arg1 cells 241:245 CD4(+) CCR5(+) host cells 221:245 CD4(+) CCR5(+) host cells 221:245 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 1 27 gly glycoprotein 175:186 arg1 Env 189:191 Env 189:191 Env 189:191 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 1 27 gly glycoprotein 175:186 arg1 glycoprotein 175:186 HIV-1 surface envelope glycoprotein 152:186 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 7 28 theme mass 949:952 arg1 spectrometry 954:965 Gel and glycan mass spectrometry 934:965 spectrometry 954:965 Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. 25253346 18 29 theme nonneutralizing 2929:2943 arg1 determinants 2945:2956 nonneutralizing determinants 2929:2956 (by glycan) nonneutralizing determinants 2917:2956 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 15 30 theme conserved 2380:2388 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 31 theme receptor 2472:2479 arg1 one 2446:2448 one 2446:2448 one 2446:2448 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 31 theme receptor 2472:2479 arg1 site 2495:2498 the virus receptor (CD4) binding site 2462:2498 the virus receptor (CD4) binding site 2462:2498 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 31 theme receptor 2472:2479 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 12 32 theme tier 1720:1723 arg1 neutralization 1733:1746 tier 1-level neutralization 1720:1746 tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent 1720:1819 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 9 33 gly hyperglycosylated 1272:1288 arg1 cores 1290:1294 the hyperglycosylated cores 1268:1294 the hyperglycosylated cores 1268:1294 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 19 34 theme structure-guided 3165:3180 arg1 modifications 3182:3194 the structure-guided modifications 3161:3194 the structure-guided modifications of the modified gp120 core 3161:3221 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 13 35 theme design 2113:2118 arg1 strategy 2120:2127 this rational design strategy 2099:2127 this rational design strategy 2099:2127 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 0 36 theme CD4 65:67 arg1 site 77:80 the CD4 binding site 61:80 the CD4 binding site 61:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 1 37 theme surface 158:164 arg1 Env 189:191 Env 189:191 Env 189:191 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 1 37 theme surface 158:164 arg1 glycoprotein 175:186 HIV-1 surface envelope glycoprotein 152:186 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 6 38 theme neutralizing 894:905 arg1 antibodies 922:931 broadly neutralizing CD4bs-directed antibodies 886:931 broadly neutralizing CD4bs-directed antibodies 886:931 Following purification, most glycoproteins, except for 10G, were recognized by broadly neutralizing CD4bs-directed antibodies. 25253346 14 39 theme Major 2142:2146 arg1 obstacles 2148:2156 Major obstacles 2142:2156 Major obstacles to developing an effective HIV-1 vaccine 2142:2197 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 19 40 theme modified 3203:3210 arg1 core 3218:3221 the modified gp120 core 3199:3221 the modified gp120 core 3199:3221 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 5 41 theme new 718:720 arg1 motifs 739:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 41 theme new 718:720 arg1 7G 788:789 7G 788:789 7G 788:789 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 41 theme new 718:720 arg1 6G 784:785 6G 784:785 6G 784:785 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 41 theme new 718:720 arg1 cores 800:804 10G cores 796:804 10G cores 796:804 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 41 theme new 718:720 arg1 3G 780:781 3G 780:781 3G 780:781 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 15 42 theme gp120 2439:2443 arg1 surface 2428:2434 the exposed envelope protein surface 2399:2434 the exposed envelope protein surface of gp120 2399:2443 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 1 43 theme glycoprotein 175:186 arg1 trimer 194:199 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 12 44 theme immunogens 1700:1709 arg1 priming 1668:1674 Trimer priming 1661:1674 Trimer priming 1661:1674 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 12 44 theme immunogens 1700:1709 arg1 boosting 1679:1686 boosting 1679:1686 boosting 1679:1686 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 15 45 from sites 2390:2394 arg1 surface 2428:2434 the exposed envelope protein surface 2399:2434 the exposed envelope protein surface of gp120 2399:2443 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 16 46 theme gp120 2583:2587 arg1 site 2601:2604 this gp120 CD4 binding site 2578:2604 this gp120 CD4 binding site 2578:2604 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 14 47 theme glycosylation 2340:2352 arg1 incorporation 2310:2322 incorporation 2310:2322 incorporation of host (human) glycosylation 2310:2352 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 9 48 theme IgM 1305:1307 arg1 responses 1317:1325 IgM and IgG responses 1305:1325 IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G 1305:1438 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 10 49 theme glycan 1455:1460 arg1 effects 1470:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 19 50 theme core 3218:3221 arg1 modifications 3182:3194 the structure-guided modifications 3161:3194 the structure-guided modifications of the modified gp120 core 3161:3221 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 0 51 theme Hyperglycosylated 0:16 arg1 immunogens 30:39 Hyperglycosylated stable core immunogens 0:39 Hyperglycosylated stable core immunogens designed to present the CD4 binding site 0:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 9 52 theme IgG 1313:1315 arg1 responses 1317:1325 IgM and IgG responses 1305:1325 IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G 1305:1438 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 15 53 theme binding 2487:2493 arg1 one 2446:2448 one 2446:2448 one 2446:2448 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 53 theme binding 2487:2493 arg1 site 2495:2498 the virus receptor (CD4) binding site 2462:2498 the virus receptor (CD4) binding site 2462:2498 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 53 theme binding 2487:2493 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 17 54 theme conformational 2728:2741 arg1 stabilization 2743:2755 conformational stabilization 2728:2755 conformational stabilization 2728:2755 In this study, we targeted the CD4bs by conformational stabilization and additional glycan masking. 25253346 14 55 theme glycan 2282:2287 arg1 variability 2211:2221 the variability 2207:2221 the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation 2207:2352 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 2 56 theme neutralization 365:378 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 56 theme neutralization 365:378 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 0 57 theme core 25:28 arg1 immunogens 30:39 Hyperglycosylated stable core immunogens 0:39 Hyperglycosylated stable core immunogens designed to present the CD4 binding site 0:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 4 58 theme additional 530:539 arg1 glycans 550:556 additional N-linked glycans 530:556 additional N-linked glycans 530:556 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 8 59 dep low 1171:1173 arg1 to 1175:1176 to 1175:1176 to 1175:1176 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 5 60 theme glycan 728:733 arg1 motifs 739:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 60 theme glycan 728:733 arg1 7G 788:789 7G 788:789 7G 788:789 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 60 theme glycan 728:733 arg1 6G 784:785 6G 784:785 6G 784:785 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 60 theme glycan 728:733 arg1 cores 800:804 10G cores 796:804 10G cores 796:804 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 60 theme glycan 728:733 arg1 3G 780:781 3G 780:781 3G 780:781 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 9 61 theme given 1335:1339 arg1 protein 1341:1347 each given protein 1330:1347 each given protein 1330:1347 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 16 62 theme Several 2501:2507 arg1 antibodies 2530:2539 Several broadly neutralizing antibodies 2501:2539 Several broadly neutralizing antibodies elicited from HIV patients 2501:2566 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 13 63 from CD4bs 1842:1846 arg1 mapping 1827:1833 Fine mapping 1822:1833 Fine mapping at the CD4bs 1822:1846 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 17 64 theme glycan 2772:2777 arg1 masking 2779:2785 additional glycan masking 2761:2785 additional glycan masking 2761:2785 In this study, we targeted the CD4bs by conformational stabilization and additional glycan masking. 25253346 19 65 theme CD4bs-directed 3065:3078 arg1 antibodies 3080:3089 the elicited CD4bs-directed antibodies 3052:3089 the elicited CD4bs-directed antibodies 3052:3089 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 2 66 theme gp120 308:312 arg1 CD4bs 349:353 CD4bs 349:353 CD4bs 349:353 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 66 theme gp120 308:312 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 66 theme gp120 308:312 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 5 67 theme G 736:736 arg1 motifs 739:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 67 theme G 736:736 arg1 7G 788:789 7G 788:789 7G 788:789 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 67 theme G 736:736 arg1 6G 784:785 6G 784:785 6G 784:785 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 67 theme G 736:736 arg1 cores 800:804 10G cores 796:804 10G cores 796:804 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 67 theme G 736:736 arg1 3G 780:781 3G 780:781 3G 780:781 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 16 68 theme neutralizing 2517:2528 arg1 antibodies 2530:2539 Several broadly neutralizing antibodies 2501:2539 Several broadly neutralizing antibodies elicited from HIV patients 2501:2566 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 16 69 theme gp120 2624:2628 arg1 immunogens 2630:2639 gp120 immunogens 2624:2639 gp120 immunogens 2624:2639 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 3 70 theme protein 427:433 arg1 surfaces 435:442 protein surfaces 427:442 protein surfaces 427:442 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 4 71 theme gp120 585:589 arg1 core 591:594 a cysteine-stabilized gp120 core 563:594 a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs 563:695 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 4 71 theme gp120 585:589 arg1 0G 597:598 0G 597:598 0G 597:598 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 13 72 theme neutralizing 1994:2005 arg1 antibody 2007:2014 neutralizing antibody 1994:2014 neutralizing antibody 1994:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 2 73 theme binding 335:341 arg1 CD4bs 349:353 CD4bs 349:353 CD4bs 349:353 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 73 theme binding 335:341 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 73 theme binding 335:341 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 15 74 theme envelope 2411:2418 arg1 surface 2428:2434 the exposed envelope protein surface 2399:2434 the exposed envelope protein surface of gp120 2399:2443 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 4 75 link N-linked 541:548 arg1 glycans 550:556 additional N-linked glycans 530:556 additional N-linked glycans 530:556 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 14 76 theme surface 2239:2245 arg1 glycoproteins 2247:2259 envelope surface glycoproteins 2230:2259 envelope surface glycoproteins 2230:2259 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 7 77 theme redesigned 1041:1050 arg1 cores 1052:1056 the redesigned cores 1037:1056 the redesigned cores 1037:1056 Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. 25253346 11 78 theme core-elicited 1602:1614 arg1 sera 1616:1619 the core-elicited sera 1598:1619 the core-elicited sera 1598:1619 However, the core-elicited sera showed limited neutralization activity. 25253346 2 79 theme receptor 322:329 arg1 CD4bs 349:353 CD4bs 349:353 CD4bs 349:353 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 79 theme receptor 322:329 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 79 theme receptor 322:329 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 16 80 theme neutralizing 2663:2674 arg1 antibodies 2676:2685 broadly neutralizing antibodies 2655:2685 broadly neutralizing antibodies 2655:2685 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 10 81 theme binding 1568:1574 arg1 specificity 1576:1586 CD4bs-directed binding specificity 1553:1586 CD4bs-directed binding specificity 1553:1586 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 11 82 theme neutralization 1636:1649 arg1 activity 1651:1658 limited neutralization activity 1628:1658 limited neutralization activity 1628:1658 However, the core-elicited sera showed limited neutralization activity. 25253346 13 83 theme sites 1968:1972 arg1 surface 1953:1959 the molecular surface 1939:1959 the molecular surface of Env sites of vulnerability to neutralizing antibody 1939:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 8 84 theme high-affinity 1085:1097 arg1 recognition 1099:1109 high-affinity recognition 1085:1109 high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies 1085:1165 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 13 85 theme vulnerability 1977:1989 arg1 sites 1968:1972 Env sites 1964:1972 Env sites of vulnerability to neutralizing antibody 1964:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 13 85 theme vulnerability 1977:1989 arg1 vulnerability 1977:1989 vulnerability 1977:1989 vulnerability to neutralizing antibody 1977:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 3 86 theme nonneutralizing 483:497 arg1 antibodies 499:508 nonneutralizing antibodies 483:508 nonneutralizing antibodies 483:508 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 4 87 theme major 616:620 arg1 regions 631:637 its major variable regions 612:637 its major variable regions to preferentially expose the conformationally fixed CD4bs 612:695 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 13 88 theme molecular 1943:1951 arg1 surface 1953:1959 the molecular surface 1939:1959 the molecular surface of Env sites of vulnerability to neutralizing antibody 1939:2014 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 8 89 theme non-broadly 1192:1202 arg1 antibodies 1232:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 14 90 theme HIV-1 2185:2189 arg1 vaccine 2191:2197 an effective HIV-1 vaccine 2172:2197 developing an effective HIV-1 vaccine 2161:2197 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 2 91 theme antigenic 272:280 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 91 theme antigenic 272:280 arg1 determinants 282:293 conserved antigenic determinants 262:293 conserved antigenic determinants 262:293 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 8 92 theme CD4bs-directed 1141:1154 arg1 antibodies 1156:1165 seven broadly neutralizing CD4bs-directed antibodies 1114:1165 seven broadly neutralizing CD4bs-directed antibodies 1114:1165 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 5 93 theme 10G 796:798 arg1 motifs 739:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 93 theme 10G 796:798 arg1 cores 800:804 10G cores 796:804 10G cores 796:804 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 8 94 theme CD4bs-directed 1217:1230 arg1 antibodies 1232:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 non-broadly neutralizing CD4bs-directed antibodies 1192:1241 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 13 95 theme cores 1899:1903 arg1 addition 1909:1916 addition 1909:1916 addition of N-glycans 1909:1929 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 13 95 theme cores 1899:1903 arg1 stabilization 1878:1890 conformational stabilization 1863:1890 conformational stabilization of the cores 1863:1903 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 18 96 used used 2791:2794 arg2 We 2788:2789 We 2788:2789 We 2788:2789 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 7 97 theme glycan 942:947 arg1 spectrometry 954:965 Gel and glycan mass spectrometry 934:965 spectrometry 954:965 Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. 25253346 18 98 theme gp120 2837:2841 arg1 cores 2843:2847 gp120 cores 2837:2847 gp120 cores 2837:2847 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 6 99 theme most 831:834 arg1 glycoproteins 836:848 most glycoproteins 831:848 most glycoproteins 831:848 Following purification, most glycoproteins, except for 10G, were recognized by broadly neutralizing CD4bs-directed antibodies. 25253346 4 100 theme fixed 685:689 arg1 CD4bs 691:695 the conformationally fixed CD4bs 664:695 the conformationally fixed CD4bs 664:695 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 7 101 theme Gel 934:936 arg1 spectrometry 954:965 Gel and glycan mass spectrometry 934:965 spectrometry 954:965 Gel and glycan mass spectrometry confirmed that additional N-glycans were posttranslationally added to the redesigned cores. 25253346 13 102 theme rational 2104:2111 arg1 design 2113:2118 this rational design 2099:2118 this rational design strategy 2099:2127 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 16 103 theme HIV 2555:2557 arg1 patients 2559:2566 HIV patients 2555:2566 HIV patients 2555:2566 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 13 104 theme N-glycans 1921:1929 arg1 addition 1909:1916 addition 1909:1916 addition of N-glycans 1909:1929 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 13 104 theme N-glycans 1921:1929 arg1 stabilization 1878:1890 conformational stabilization 1863:1890 conformational stabilization of the cores 1863:1903 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 0 105 theme binding 69:75 arg1 site 77:80 the CD4 binding site 61:80 the CD4 binding site 61:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 15 106 theme virus 2466:2470 arg1 one 2446:2448 one 2446:2448 one 2446:2448 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 106 theme virus 2466:2470 arg1 site 2495:2498 the virus receptor (CD4) binding site 2462:2498 the virus receptor (CD4) binding site 2462:2498 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 15 106 theme virus 2466:2470 arg1 sites 2390:2394 highly conserved sites 2373:2394 highly conserved sites 2373:2394 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 1 107 theme HIV-1 152:156 arg1 Env 189:191 Env 189:191 Env 189:191 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 1 107 theme HIV-1 152:156 arg1 glycoprotein 175:186 HIV-1 surface envelope glycoprotein 152:186 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 12 108 theme 1-level 1725:1731 arg1 neutralization 1733:1746 tier 1-level neutralization 1720:1746 tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent 1720:1819 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 6 109 theme CD4bs-directed 907:920 arg1 antibodies 922:931 broadly neutralizing CD4bs-directed antibodies 886:931 broadly neutralizing CD4bs-directed antibodies 886:931 Following purification, most glycoproteins, except for 10G, were recognized by broadly neutralizing CD4bs-directed antibodies. 25253346 14 110 theme host 2327:2330 arg1 glycosylation 2340:2352 host (human) glycosylation 2327:2352 host (human) glycosylation 2327:2352 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 14 111 theme human 2333:2337 arg1 glycosylation 2340:2352 host (human) glycosylation 2327:2352 host (human) glycosylation 2327:2352 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 1 112 theme envelope 166:173 arg1 Env 189:191 Env 189:191 Env 189:191 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 1 112 theme envelope 166:173 arg1 glycoprotein 175:186 HIV-1 surface envelope glycoprotein 152:186 The HIV-1 surface envelope glycoprotein (Env) trimer 148:199 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 0 113 theme neutralizing 123:134 arg1 antibodies 136:145 broadly neutralizing antibodies 115:145 broadly neutralizing antibodies 115:145 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 10 114 located detected 1483:1490 arg2 effects 1470:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 10 114 located detected 1483:1490 arg1 sera 1508:1511 the elicited sera 1495:1511 the elicited sera 1495:1511 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 12 115 dep CD4bs 1772:1776 arg1 the 1768:1770 the 1768:1770 the 1768:1770 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 9 116 theme hyperglycosylated 1272:1288 arg1 cores 1290:1294 the hyperglycosylated cores 1268:1294 the hyperglycosylated cores 1268:1294 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 12 117 theme core 1695:1698 arg1 immunogens 1700:1709 the core immunogens 1691:1709 the core immunogens 1691:1709 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 16 118 theme CD4 2589:2591 arg1 site 2601:2604 this gp120 CD4 binding site 2578:2604 this gp120 CD4 binding site 2578:2604 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 18 119 theme cysteine-stabilized 2879:2897 arg1 CD4bs 2899:2903 the cysteine-stabilized CD4bs 2875:2903 the cysteine-stabilized CD4bs 2875:2903 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 12 120 theme Trimer 1661:1666 arg1 priming 1668:1674 Trimer priming 1661:1674 Trimer priming 1661:1674 Trimer priming or boosting of the core immunogens elicited tier 1-level neutralization that mapped to both the CD4bs and V3 and appeared to be trimer dependent. 25253346 14 121 dep glycoproteins 2247:2259 arg1 the 2226:2228 the 2226:2228 the 2226:2228 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 10 122 theme masking 1462:1468 arg1 effects 1470:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 13 123 theme conformational 1863:1876 arg1 stabilization 1878:1890 conformational stabilization 1863:1890 conformational stabilization of the cores 1863:1903 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 0 124 theme stable 18:23 arg1 immunogens 30:39 Hyperglycosylated stable core immunogens 0:39 Hyperglycosylated stable core immunogens designed to present the CD4 binding site 0:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 15 125 theme protein 2420:2426 arg1 surface 2428:2434 the exposed envelope protein surface 2399:2434 the exposed envelope protein surface of gp120 2399:2443 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 10 126 theme Site-specific 1441:1453 arg1 effects 1470:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects 1441:1476 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 15 127 theme exposed 2403:2409 arg1 surface 2428:2434 the exposed envelope protein surface 2399:2434 the exposed envelope protein surface of gp120 2399:2443 HIV-1 does harbor highly conserved sites on the exposed envelope protein surface of gp120, one of which is the virus receptor (CD4) binding site. 25253346 4 128 theme N-linked 541:548 arg1 glycans 550:556 additional N-linked glycans 530:556 additional N-linked glycans 530:556 Here we engineered additional N-linked glycans onto a cysteine-stabilized gp120 core (0G) deleted of its major variable regions to preferentially expose the conformationally fixed CD4bs. 25253346 14 129 theme high-density 2269:2280 arg1 glycan 2282:2287 its high-density glycan shield 2265:2294 its high-density glycan shield 2265:2294 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 2 130 theme known 359:363 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 130 theme known 359:363 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 5 131 theme NXT/S 722:726 arg1 motifs 739:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs 698:744 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 131 theme NXT/S 722:726 arg1 7G 788:789 7G 788:789 7G 788:789 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 131 theme NXT/S 722:726 arg1 6G 784:785 6G 784:785 6G 784:785 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 131 theme NXT/S 722:726 arg1 cores 800:804 10G cores 796:804 10G cores 796:804 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 5 131 theme NXT/S 722:726 arg1 3G 780:781 3G 780:781 3G 780:781 Three, 6, 7, and 10 new NXT/S glycan (G) motifs were engineered into 0G to encode 3G, 6G, 7G, and 10G cores. 25253346 2 132 contain possesses 252:260 arg1 Env 248:250 Env 248:250 Env 248:250 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 132 contain possesses 252:260 arg2 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 132 contain possesses 252:260 arg2 determinants 282:293 conserved antigenic determinants 262:293 conserved antigenic determinants 262:293 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 9 133 from characteristics 1391:1405 arg1 similar 1359:1365 similar 1359:1365 similar 1359:1365 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 0 134 gly Hyperglycosylated 0:16 arg1 immunogens 30:39 Hyperglycosylated stable core immunogens 0:39 Hyperglycosylated stable core immunogens designed to present the CD4 binding site 0:80 Hyperglycosylated stable core immunogens designed to present the CD4 binding site are preferentially recognized by broadly neutralizing antibodies. 25253346 18 135 dep used 2791:2794 arg1 mask 2912:2915 mask 2912:2915 to mask (by glycan) nonneutralizing determinants 2909:2956 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 18 135 dep used 2791:2794 arg1 reengineer 2826:2835 reengineer 2826:2835 to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs 2823:2903 We used the atomic-level structure to reengineer gp120 cores to preferentially present the cysteine-stabilized CD4bs and to mask (by glycan) nonneutralizing determinants. 25253346 17 136 theme additional 2761:2770 arg1 masking 2779:2785 additional glycan masking 2761:2785 additional glycan masking 2761:2785 In this study, we targeted the CD4bs by conformational stabilization and additional glycan masking. 25253346 19 137 theme antibody 3010:3017 arg1 responses 3019:3027 antibody responses 3010:3027 antibody responses to the CD4bs 3010:3040 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 19 138 theme elicited 3056:3063 arg1 antibodies 3080:3089 the elicited CD4bs-directed antibodies 3052:3089 the elicited CD4bs-directed antibodies 3052:3089 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 3 139 theme variable 406:413 arg1 regions 415:421 variable regions 406:421 variable regions 406:421 Env also contains variable regions and protein surfaces occluded within the trimer that elicit nonneutralizing antibodies. 25253346 13 140 theme elicited 2055:2062 arg1 neutralization 2064:2077 the elicited neutralization 2051:2077 the elicited neutralization 2051:2077 Fine mapping at the CD4bs indicated that conformational stabilization of the cores and addition of N-glycans altered the molecular surface of Env sites of vulnerability to neutralizing antibody, suggesting an explanation for why the elicited neutralization was not improved by this rational design strategy. 25253346 9 141 from similar 1359:1365 arg1 characteristics 1391:1405 their neutralization characteristics 1370:1405 their neutralization characteristics 1370:1405 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 25253346 16 142 theme broadly 2509:2515 arg1 antibodies 2530:2539 Several broadly neutralizing antibodies 2501:2539 Several broadly neutralizing antibodies elicited from HIV patients 2501:2566 Several broadly neutralizing antibodies elicited from HIV patients do target this gp120 CD4 binding site (CD4bs); however, gp120 immunogens do not elicit broadly neutralizing antibodies. 25253346 10 143 theme elicited 1499:1506 arg1 sera 1508:1511 the elicited sera 1495:1511 the elicited sera 1495:1511 Site-specific glycan masking effects were detected in the elicited sera, and the antisera competed with b12 for CD4bs-directed binding specificity. 25253346 19 144 theme gp120 3212:3216 arg1 core 3218:3221 the modified gp120 core 3199:3221 the modified gp120 core 3199:3221 Importantly, glycan masking did successfully focus antibody responses to the CD4bs; however, the elicited CD4bs-directed antibodies did not neutralize HIV or bind to unmodified gp120, presumably due to the structure-guided modifications of the modified gp120 core. 25253346 8 145 theme Binding 1059:1065 arg1 kinetics 1067:1074 Binding kinetics 1059:1074 Binding kinetics 1059:1074 Binding kinetics revealed high-affinity recognition by seven broadly neutralizing CD4bs-directed antibodies and low to no binding by non-broadly neutralizing CD4bs-directed antibodies. 25253346 14 146 gly glycoproteins 2247:2259 arg1 glycoproteins 2247:2259 envelope surface glycoproteins 2230:2259 envelope surface glycoproteins 2230:2259 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 14 147 theme glycoproteins 2247:2259 arg1 variability 2211:2221 the variability 2207:2221 the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation 2207:2352 IMPORTANCE: Major obstacles to developing an effective HIV-1 vaccine include the variability of the envelope surface glycoproteins and its high-density glycan shield, generated by incorporation of host (human) glycosylation. 25253346 1 148 theme CCR5 228:231 arg1 cells 241:245 CD4(+) CCR5(+) host cells 221:245 CD4(+) CCR5(+) host cells 221:245 The HIV-1 surface envelope glycoprotein (Env) trimer mediates entry into CD4(+) CCR5(+) host cells. 25253346 2 149 theme CD4 331:333 arg1 CD4bs 349:353 CD4bs 349:353 CD4bs 349:353 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 149 theme CD4 331:333 arg1 site 343:346 the gp120 primary receptor CD4 binding site 304:346 the gp120 primary receptor CD4 binding site (CD4bs) 304:354 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 2 149 theme CD4 331:333 arg1 target 380:385 a known neutralization target 357:385 a known neutralization target 357:385 Env possesses conserved antigenic determinants, such as the gp120 primary receptor CD4 binding site (CD4bs), a known neutralization target. 25253346 9 150 theme neutralization 1376:1389 arg1 characteristics 1391:1405 their neutralization characteristics 1370:1405 their neutralization characteristics 1370:1405 Rabbits inoculated with the hyperglycosylated cores elicited IgM and IgG responses to each given protein that were similar in their neutralization characteristics to those elicited by parental 0G. 24533768 5 0 theme Periodate 716:724 arg1 oxidation 726:734 Periodate oxidation 716:734 Periodate oxidation of these sialic acids 716:756 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 3 1 from site 494:497 arg1 antibodies 502:511 antibodies 502:511 antibodies 502:511 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 1 2 theme native 171:176 arg1 antibodies 178:187 native antibodies 171:187 native antibodies 171:187 Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. 24533768 7 3 theme residues 1175:1182 arg1 number 1153:1158 the number 1149:1158 the number of sialic acid residues 1149:1182 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 8 4 theme antitumor 1329:1337 arg1 efficacy 1339:1346 significantly greater antitumor efficacy 1307:1346 significantly greater antitumor efficacy 1307:1346 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 4 5 theme native 562:567 arg1 glycans 569:575 The native glycans 558:575 The native glycans on Asn-297 of antibodies 558:600 The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. 24533768 9 6 theme oxime 1473:1477 arg1 ligation 1479:1486 oxime ligation 1473:1486 oxime ligation of the native glycans of antibodies 1473:1522 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 7 7 theme cytotoxic 1095:1103 arg1 agents 1105:1110 ~1.6 cytotoxic agents 1090:1110 ~1.6 cytotoxic agents 1090:1110 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 8 8 theme naked 1353:1357 arg1 antibody 1359:1366 naked antibody 1353:1366 naked antibody 1353:1366 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 9 9 theme glycans 1502:1508 arg1 ligation 1479:1486 oxime ligation 1473:1486 oxime ligation of the native glycans of antibodies 1473:1522 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 7 10 theme agents 1105:1110 arg1 incorporation 1073:1085 the incorporation 1069:1085 the incorporation of ~1.6 cytotoxic agents per antibody molecule 1069:1132 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 7 11 theme antibody 1116:1123 arg1 molecule 1125:1132 antibody molecule 1116:1132 antibody molecule 1116:1132 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 10 12 theme controlled 1794:1803 arg1 process 1814:1820 a highly controlled chemical process 1785:1820 a highly controlled chemical process to ensure reproducible drug loading 1785:1856 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 5 13 used used 806:809 arg2 groups 775:780 aldehyde groups 766:780 aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation 766:882 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 8 14 theme tumor 1291:1295 arg1 cells 1297:1301 antigen-positive tumor cells 1274:1301 antigen-positive tumor cells 1274:1301 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 2 15 theme product 355:361 arg1 profiles 363:370 heterogeneous product profiles 341:370 heterogeneous product profiles 341:370 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 10 16 theme reproducible 1832:1843 arg1 loading 1850:1856 reproducible drug loading 1832:1856 reproducible drug loading 1832:1856 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 9 17 theme antibodies 1513:1522 arg1 glycans 1502:1508 the native glycans 1491:1508 the native glycans of antibodies 1491:1522 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 8 18 theme glyco-conjugated 1191:1206 arg1 ADCs 1208:1211 These glyco-conjugated ADCs 1185:1211 These glyco-conjugated ADCs 1185:1211 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 7 19 theme Hydrophobic 1002:1012 arg1 chromatography 1026:1039 Hydrophobic interaction chromatography 1002:1039 Hydrophobic interaction chromatography 1002:1039 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 10 20 theme viable 1672:1677 arg1 alternative 1679:1689 a viable alternative 1670:1689 a viable alternative 1670:1689 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 3 21 theme glycosylation 480:492 arg1 approach 519:526 an approach 516:526 an approach to address these limitations 516:555 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 3 21 theme glycosylation 480:492 arg1 site 494:497 the native glycosylation site 469:497 the native glycosylation site on antibodies 469:511 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 5 22 theme aminooxy 824:831 arg1 agents 858:863 aminooxy functionalized cytotoxic agents 824:863 aminooxy functionalized cytotoxic agents 824:863 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 0 23 theme Site-specific 0:12 arg1 antibody-drug 14:26 Site-specific antibody-drug 0:26 Site-specific antibody-drug 0:26 Site-specific antibody-drug conjugation through glycoengineering. 24533768 3 24 theme novel 417:421 arg1 method 449:454 a novel site-specific conjugation method 415:454 a novel site-specific conjugation method 415:454 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 6 25 theme cytotoxic 984:992 arg1 trastuzumab 964:974 trastuzumab 964:974 trastuzumab 964:974 The process has been successfully demonstrated with three antibodies including trastuzumab and two cytotoxic agents. 24533768 6 25 theme cytotoxic 984:992 arg1 agents 994:999 two cytotoxic agents 980:999 two cytotoxic agents 980:999 The process has been successfully demonstrated with three antibodies including trastuzumab and two cytotoxic agents. 24533768 10 26 theme other 1694:1698 arg1 methods 1700:1706 other methods 1694:1706 other methods 1694:1706 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 8 27 theme greater 1321:1327 arg1 efficacy 1339:1346 significantly greater antitumor efficacy 1307:1346 significantly greater antitumor efficacy 1307:1346 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 2 28 theme heterogeneous 341:353 arg1 profiles 363:370 heterogeneous product profiles 341:370 heterogeneous product profiles 341:370 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 8 29 theme antigen-positive 1274:1289 arg1 cells 1297:1301 antigen-positive tumor cells 1274:1301 antigen-positive tumor cells 1274:1301 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 3 30 theme site-specific 423:435 arg1 method 449:454 a novel site-specific conjugation method 415:454 a novel site-specific conjugation method 415:454 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 9 31 theme attractive 1534:1543 arg1 approach 1545:1552 an attractive approach 1531:1552 an attractive approach to generate ADCs with well-defined product profiles 1531:1604 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 9 32 with ADCs 1566:1569 arg1 profiles 1597:1604 well-defined product profiles 1576:1604 well-defined product profiles 1576:1604 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 3 33 theme native 473:478 arg1 approach 519:526 an approach 516:526 an approach to address these limitations 516:555 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 3 33 theme native 473:478 arg1 site 494:497 the native glycosylation site 469:497 the native glycosylation site on antibodies 469:511 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 8 34 theme xenograft 1393:1401 arg1 model 1403:1407 a Her2-positive tumor xenograft model 1371:1407 a Her2-positive tumor xenograft model 1371:1407 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 2 35 contain have 301:304 arg2 number 308:313 a number 306:313 a number of shortcomings including heterogeneous product profiles and linkage instability 306:394 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 2 35 contain have 301:304 arg1 chemistries 225:235 the classical conjugation chemistries 199:235 the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides 199:299 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 2 35 contain have 301:304 arg2 instability 384:394 linkage instability 376:394 linkage instability 376:394 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 2 35 contain have 301:304 arg2 profiles 363:370 heterogeneous product profiles 341:370 heterogeneous product profiles 341:370 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 8 36 theme antiproliferative 1240:1256 arg1 activity 1258:1265 target-dependent antiproliferative activity 1223:1265 target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody 1223:1366 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 10 37 theme conjugation 1625:1635 arg1 approach 1637:1644 The site-specific conjugation approach 1607:1644 The site-specific conjugation approach presented here 1607:1659 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 2 38 theme shortcomings 318:329 arg1 number 308:313 a number 306:313 a number of shortcomings including heterogeneous product profiles and linkage instability 306:394 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 2 38 theme shortcomings 318:329 arg1 profiles 363:370 heterogeneous product profiles 341:370 heterogeneous product profiles 341:370 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 2 38 theme shortcomings 318:329 arg1 instability 384:394 linkage instability 376:394 linkage instability 376:394 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 10 39 theme site-specific 1611:1623 arg1 approach 1637:1644 The site-specific conjugation approach 1607:1644 The site-specific conjugation approach presented here 1607:1659 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 4 40 theme sialic 702:707 arg1 acids 709:713 terminal sialic acids 693:713 terminal sialic acids 693:713 The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. 24533768 8 41 theme tumor 1387:1391 arg1 model 1403:1407 a Her2-positive tumor xenograft model 1371:1407 a Her2-positive tumor xenograft model 1371:1407 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 8 42 theme target-dependent 1223:1238 arg1 activity 1258:1265 target-dependent antiproliferative activity 1223:1265 target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody 1223:1366 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 7 43 theme acid 1170:1173 arg1 residues 1175:1182 sialic acid residues 1163:1182 sialic acid residues 1163:1182 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 4 44 theme terminal 693:700 arg1 acids 709:713 terminal sialic acids 693:713 terminal sialic acids 693:713 The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. 24533768 5 45 theme oxime 869:873 arg1 ligation 875:882 oxime ligation 869:882 oxime ligation 869:882 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 10 46 theme drug 1845:1848 arg1 loading 1850:1856 reproducible drug loading 1832:1856 reproducible drug loading 1832:1856 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 9 47 theme enzymatic 1438:1446 arg1 remodeling 1448:1457 enzymatic remodeling 1438:1457 enzymatic remodeling combined with oxime ligation of the native glycans of antibodies 1438:1522 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 3 48 theme conjugation 437:447 arg1 method 449:454 a novel site-specific conjugation method 415:454 a novel site-specific conjugation method 415:454 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 10 49 theme chemical 1805:1812 arg1 process 1814:1820 a highly controlled chemical process 1785:1820 a highly controlled chemical process to ensure reproducible drug loading 1785:1856 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 5 50 theme sialic 745:750 arg1 acids 752:756 these sialic acids 739:756 these sialic acids 739:756 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 4 51 theme antibodies 591:600 arg1 Asn-297 580:586 Asn-297 580:586 Asn-297 of antibodies 580:600 The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. 24533768 10 52 dep sequence 1765:1772 arg1 either 1733:1738 either 1733:1738 either 1733:1738 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 10 52 dep sequence 1765:1772 arg1 re-engineer 1740:1750 re-engineer 1740:1750 re-engineer 1740:1750 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 2 53 theme linkage 376:382 arg1 instability 384:394 linkage instability 376:394 linkage instability 376:394 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 4 54 from glycans 569:575 arg1 Asn-297 580:586 Asn-297 580:586 Asn-297 of antibodies 580:600 The native glycans on Asn-297 of antibodies were enzymatically remodeled in vitro using galactosyl and sialyltransferases to introduce terminal sialic acids. 24533768 9 55 theme well-defined 1576:1587 arg1 profiles 1597:1604 well-defined product profiles 1576:1604 well-defined product profiles 1576:1604 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 5 56 theme acids 752:756 arg1 oxidation 726:734 Periodate oxidation 716:734 Periodate oxidation of these sialic acids 716:756 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 9 57 theme product 1589:1595 arg1 profiles 1597:1604 well-defined product profiles 1576:1604 well-defined product profiles 1576:1604 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 7 58 theme sialic 1163:1168 arg1 residues 1175:1182 sialic acid residues 1163:1182 sialic acid residues 1163:1182 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 2 59 theme hinge 284:288 arg1 disulfides 290:299 hinge disulfides 284:299 hinge disulfides 284:299 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 1 60 theme effective 137:145 arg1 agents 159:164 more effective anti-cancer agents 132:164 more effective anti-cancer agents 132:164 Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. 24533768 7 61 theme LC-MS 1045:1049 arg1 analyses 1051:1058 LC-MS analyses 1045:1058 LC-MS analyses 1045:1058 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 5 62 theme aldehyde 766:773 arg1 groups 775:780 aldehyde groups 766:780 aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation 766:882 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 2 63 theme conjugation 213:223 arg1 chemistries 225:235 the classical conjugation chemistries 199:235 the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides 199:299 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 3 64 gly glycosylation 480:492 arg2 approach 519:526 an approach 516:526 an approach to address these limitations 516:555 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 3 64 gly glycosylation 480:492 arg2 site 494:497 the native glycosylation site 469:497 the native glycosylation site on antibodies 469:511 We have developed a novel site-specific conjugation method by targeting the native glycosylation site on antibodies as an approach to address these limitations. 24533768 1 65 theme anti-cancer 147:157 arg1 agents 159:164 more effective anti-cancer agents 132:164 more effective anti-cancer agents 132:164 Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. 24533768 7 66 theme interaction 1014:1024 arg1 chromatography 1026:1039 Hydrophobic interaction chromatography 1002:1039 Hydrophobic interaction chromatography 1002:1039 Hydrophobic interaction chromatography and LC-MS analyses revealed the incorporation of ~1.6 cytotoxic agents per antibody molecule, approximating the number of sialic acid residues. 24533768 5 67 theme functionalized 833:846 arg1 agents 858:863 aminooxy functionalized cytotoxic agents 824:863 aminooxy functionalized cytotoxic agents 824:863 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 2 68 theme classical 203:211 arg1 chemistries 225:235 the classical conjugation chemistries 199:235 the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides 199:299 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 1 69 theme Antibody-drug 66:78 arg1 ADCs 92:95 ADCs 92:95 ADCs 92:95 Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. 24533768 1 69 theme Antibody-drug 66:78 arg1 conjugates 80:89 Antibody-drug conjugates 66:89 Antibody-drug conjugates (ADCs) 66:96 Antibody-drug conjugates (ADCs) have been proven clinically to be more effective anti-cancer agents than native antibodies. 24533768 10 70 theme antibody 1756:1763 arg1 sequence 1765:1772 the antibody sequence 1752:1772 the antibody sequence 1752:1772 The site-specific conjugation approach presented here provides a viable alternative to other methods, which involve a need to either re-engineer the antibody sequence or develop a highly controlled chemical process to ensure reproducible drug loading. 24533768 5 71 theme cytotoxic 848:856 arg1 agents 858:863 aminooxy functionalized cytotoxic agents 824:863 aminooxy functionalized cytotoxic agents 824:863 Periodate oxidation of these sialic acids yielded aldehyde groups which were subsequently used to conjugate aminooxy functionalized cytotoxic agents via oxime ligation. 24533768 9 72 theme native 1495:1500 arg1 glycans 1502:1508 the native glycans 1491:1508 the native glycans of antibodies 1491:1522 These findings suggest that enzymatic remodeling combined with oxime ligation of the native glycans of antibodies offers an attractive approach to generate ADCs with well-defined product profiles. 24533768 2 73 theme primary 266:272 arg1 amines 274:279 primary amines 266:279 primary amines 266:279 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24533768 8 74 theme Her2-positive 1373:1385 arg1 model 1403:1407 a Her2-positive tumor xenograft model 1371:1407 a Her2-positive tumor xenograft model 1371:1407 These glyco-conjugated ADCs exhibited target-dependent antiproliferative activity toward antigen-positive tumor cells and significantly greater antitumor efficacy than naked antibody in a Her2-positive tumor xenograft model. 24533768 2 75 dep chemistries 225:235 arg1 prepare 240:246 prepare 240:246 to prepare ADCs by targeting primary amines or hinge disulfides 237:299 However, the classical conjugation chemistries to prepare ADCs by targeting primary amines or hinge disulfides have a number of shortcomings including heterogeneous product profiles and linkage instability. 24941220 8 0 theme 2224 1217:1220 arg1 spectra 1222:1228 2224 spectra 1217:1228 2224 spectra using only 18 spectral templates 1217:1261 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 6 1 theme glycosylation 957:969 arg1 glycosylation 957:969 glycosylation 957:969 glycosylation 957:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 6 1 theme glycosylation 957:969 arg1 site 949:952 the site 945:952 the site of glycosylation 945:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 4 2 theme low 608:610 arg1 glycopeptides 621:633 low abundant glycopeptides 608:633 low abundant glycopeptides from virion-associated gp120 608:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 6 3 theme MS/MS 885:889 arg1 pattern 891:897 very similar MS/MS pattern 872:897 very similar MS/MS pattern 872:897 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 12 4 contain has 1908:1910 arg2 implications 1912:1923 implications 1912:1923 implications 1912:1923 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 12 4 contain has 1908:1910 arg1 observation 1841:1851 This direct observation 1829:1851 This direct observation of site-specific glycosylation of virion-derived gp120 1829:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 1 5 theme immunodeficiency 197:212 arg1 virus 214:218 human immunodeficiency virus 191:218 human immunodeficiency virus (HIV) 191:224 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 1 5 theme immunodeficiency 197:212 arg1 HIV 221:223 HIV 221:223 HIV 221:223 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 5 6 theme HCD-MS 679:684 arg1 data 686:689 the HCD-MS data 675:689 the HCD-MS data for glycopeptides 675:707 To search the HCD-MS data for glycopeptides, a novel spectral-aligning strategy was developed. 24941220 6 7 theme corresponding 828:840 arg1 peptides 857:864 the corresponding deglycosylated peptides 824:864 the corresponding deglycosylated peptides 824:864 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 8 8 link O-linked 1396:1403 arg1 glycosites 1426:1435 glycosites 1426:1435 glycosites 1426:1435 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 8 link O-linked 1396:1403 arg1 sites 1419:1423 one O-linked glycosylation sites 1392:1423 one O-linked glycosylation sites (glycosites) 1392:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 10 9 gly glycosites 1697:1706 arg2 glycosites 1697:1706 an additional four N-linked glycosites 1669:1706 an additional four N-linked glycosites 1669:1706 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 8 10 theme precursor 1274:1282 arg1 correction 1289:1298 precursor mass correction 1274:1298 precursor mass correction 1274:1298 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 1 11 dep glycan 250:255 arg1 shield 257:262 shield 257:262 shield 257:262 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 6 12 contain contain 937:943 arg2 glycosylation 957:969 glycosylation 957:969 glycosylation 957:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 6 12 contain contain 937:943 arg1 y-ions 918:923 y-ions 918:923 y-ions 918:923 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 6 12 contain contain 937:943 arg2 site 949:952 the site 945:952 the site of glycosylation 945:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 6 12 contain contain 937:943 arg1 b- 911:912 b- 911:912 b- 911:912 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 9 13 theme native 1514:1519 arg1 gp120 1521:1525 native gp120 1514:1525 native gp120 1514:1525 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 5 14 theme spectral-aligning 718:734 arg1 strategy 736:743 a novel spectral-aligning strategy 710:743 a novel spectral-aligning strategy 710:743 To search the HCD-MS data for glycopeptides, a novel spectral-aligning strategy was developed. 24941220 10 15 theme high 1628:1631 arg1 mannose 1633:1639 high mannose 1628:1639 high mannose 1628:1639 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 5 16 theme novel 712:716 arg1 strategy 736:743 a novel spectral-aligning strategy 710:743 a novel spectral-aligning strategy 710:743 To search the HCD-MS data for glycopeptides, a novel spectral-aligning strategy was developed. 24941220 8 17 theme unique 1341:1346 arg1 glycopeptides 1348:1360 460 unique glycopeptides 1337:1360 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites) 1337:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 7 18 theme resembling 1043:1052 arg1 spectra 1054:1060 nearly resembling spectra 1036:1060 nearly resembling spectra regardless of the attached glycan structures 1036:1105 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 11 19 theme O-linked 1754:1761 arg1 Gal1GalNAc1 1770:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 4 20 from gp120 658:662 arg1 glycopeptides 621:633 low abundant glycopeptides 608:633 low abundant glycopeptides from virion-associated gp120 608:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 1 21 theme protein 174:180 arg1 gp120 182:186 Envelope protein gp120 165:186 Envelope protein gp120 of human immunodeficiency virus (HIV) 165:224 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 7 22 theme peptide 1014:1020 arg1 backbone 1022:1029 an identical peptide backbone 1001:1029 an identical peptide backbone 1001:1029 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 12 23 theme virion-derived 1887:1900 arg1 gp120 1902:1906 virion-derived gp120 1887:1906 virion-derived gp120 1887:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 0 24 theme higher 94:99 arg1 dissociation 120:131 higher energy collisional dissociation and spectral-aligning strategy 94:162 dissociation 120:131 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 8 25 gly glycopeptides 1348:1360 arg2 glycopeptides 1348:1360 460 unique glycopeptides 1337:1360 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites) 1337:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 11 26 link O-linked 1799:1806 arg1 glycosite 1808:1816 the O-linked glycosite 1795:1816 the O-linked glycosite 1795:1816 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 8 27 theme glycosylation 1405:1417 arg1 glycosites 1426:1435 glycosites 1426:1435 glycosites 1426:1435 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 27 theme glycosylation 1405:1417 arg1 sites 1419:1423 one O-linked glycosylation sites 1392:1423 one O-linked glycosylation sites (glycosites) 1392:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 10 28 theme additional 1672:1681 arg1 glycosites 1697:1706 an additional four N-linked glycosites 1669:1706 an additional four N-linked glycosites 1669:1706 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 0 29 theme collisional 108:118 arg1 dissociation 120:131 higher energy collisional dissociation and spectral-aligning strategy 94:162 dissociation 120:131 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 12 30 theme HIV 1928:1930 arg1 glycobiology 1932:1943 HIV glycobiology 1928:1943 HIV glycobiology 1928:1943 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 1 31 theme envelope 295:302 arg1 folding 304:310 envelope folding 295:310 envelope folding 295:310 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 2 32 theme recombinant 403:413 arg1 gp120 415:419 recombinant gp120 403:419 recombinant gp120 403:419 Site-specific glycosylation profiling of recombinant gp120 is very challenging. 24941220 0 33 theme Glycoform 0:8 arg1 analysis 10:17 Glycoform analysis 0:17 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 0:163 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 1 34 theme critical 277:284 arg1 roles 286:290 critical roles 277:290 critical roles 277:290 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 10 35 from Asn276 1564:1569 arg1 region 1581:1586 the C2 region 1574:1586 the C2 region 1574:1586 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 4 36 theme Q-Exactive 567:576 arg1 spectrometer 583:594 a Q-Exactive mass spectrometer 565:594 a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120 565:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 12 37 gly glycosylation 1870:1882 arg1 gp120 1902:1906 virion-derived gp120 1887:1906 virion-derived gp120 1887:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 3 38 theme gp120 493:497 arg1 analysis 468:475 glycoproteomic analysis 453:475 glycoproteomic analysis of native viral gp120 453:497 Therefore, glycoproteomic analysis of native viral gp120 is still formidable to date. 24941220 12 39 theme gp120 1902:1906 arg1 glycosylation 1870:1882 site-specific glycosylation 1856:1882 site-specific glycosylation of virion-derived gp120 1856:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 10 40 theme N-linked 1688:1695 arg1 glycosites 1697:1706 an additional four N-linked glycosites 1669:1706 an additional four N-linked glycosites 1669:1706 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 8 41 gly glycopeptides 1173:1185 arg2 glycopeptides 1173:1185 glycopeptides 1173:1185 glycopeptides 1173:1185 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 3 42 theme native 480:485 arg1 gp120 493:497 native viral gp120 480:497 native viral gp120 480:497 Therefore, glycoproteomic analysis of native viral gp120 is still formidable to date. 24941220 9 43 theme five 1475:1478 arg1 N- 1480:1481 N- 1480:1481 N- 1480:1481 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 8 44 theme copy-paste 1141:1150 arg1 pattern 1162:1168 this "copy-paste" spectral pattern 1135:1168 this "copy-paste" spectral pattern of glycopeptides 1135:1185 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 12 45 theme direct 1834:1839 arg1 observation 1841:1851 This direct observation 1829:1851 This direct observation of site-specific glycosylation of virion-derived gp120 1829:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 0 46 theme envelope 67:74 arg1 gp120 84:88 envelope protein gp120 67:88 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 10 47 theme mannose 1733:1739 arg1 type 1741:1744 high mannose type 1728:1744 high mannose type 1728:1744 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 0 48 theme immunodeficiency 44:59 arg1 virus 61:65 recombinant and human immunodeficiency virus 22:65 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 8 49 gly glycosites 1426:1435 arg2 glycosites 1426:1435 glycosites 1426:1435 glycosites 1426:1435 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 49 gly glycosites 1426:1435 arg2 sites 1419:1423 one O-linked glycosylation sites 1392:1423 one O-linked glycosylation sites (glycosites) 1392:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 50 theme spectral 1153:1160 arg1 pattern 1162:1168 this "copy-paste" spectral pattern 1135:1168 this "copy-paste" spectral pattern of glycopeptides 1135:1185 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 7 51 gly glycopeptides 982:994 arg2 glycopeptides 982:994 glycopeptides 982:994 glycopeptides with an identical peptide backbone 982:1029 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 10 52 theme C2 1578:1579 arg1 region 1581:1586 the C2 region 1574:1586 the C2 region 1574:1586 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 9 53 theme N- 1480:1481 arg1 glycosites 1500:1509 five N- and one O-linked glycosites 1475:1509 five N- and one O-linked glycosites 1475:1509 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 1 54 theme dense 244:248 arg1 glycan 250:255 a dense glycan shield 242:262 a dense glycan shield 242:262 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 2 55 theme Site-specific 362:374 arg1 profiling 390:398 Site-specific glycosylation profiling 362:398 Site-specific glycosylation profiling of recombinant gp120 362:419 Site-specific glycosylation profiling of recombinant gp120 is very challenging. 24941220 6 56 theme similar 877:883 arg1 pattern 891:897 very similar MS/MS pattern 872:897 very similar MS/MS pattern 872:897 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 8 57 theme spectra 1222:1228 arg1 identification 1199:1212 identification 1199:1212 identification of 2224 spectra using only 18 spectral templates 1199:1261 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 4 58 theme abundant 612:619 arg1 glycopeptides 621:633 low abundant glycopeptides 608:633 low abundant glycopeptides from virion-associated gp120 608:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 1 59 theme human 191:195 arg1 virus 214:218 human immunodeficiency virus 191:218 human immunodeficiency virus (HIV) 191:224 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 1 59 theme human 191:195 arg1 HIV 221:223 HIV 221:223 HIV 221:223 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 6 60 theme deglycosylated 842:855 arg1 peptides 857:864 the corresponding deglycosylated peptides 824:864 the corresponding deglycosylated peptides 824:864 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 12 61 theme site-specific 1856:1868 arg1 glycosylation 1870:1882 site-specific glycosylation 1856:1882 site-specific glycosylation of virion-derived gp120 1856:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 1 62 theme virus 214:218 arg1 gp120 182:186 Envelope protein gp120 165:186 Envelope protein gp120 of human immunodeficiency virus (HIV) 165:224 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 8 63 theme spectral 1244:1251 arg1 templates 1253:1261 only 18 spectral templates 1236:1261 only 18 spectral templates 1236:1261 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 6 64 gly deglycosylated 842:855 arg1 peptides 857:864 the corresponding deglycosylated peptides 824:864 the corresponding deglycosylated peptides 824:864 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 11 65 theme Core 1747:1750 arg1 Gal1GalNAc1 1770:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 5 66 gly glycopeptides 695:707 arg2 glycopeptides 695:707 glycopeptides 695:707 glycopeptides 695:707 To search the HCD-MS data for glycopeptides, a novel spectral-aligning strategy was developed. 24941220 0 67 theme spectral-aligning 137:153 arg1 strategy 155:162 higher energy collisional dissociation and spectral-aligning strategy 94:162 strategy 155:162 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 9 68 theme O-linked 1491:1498 arg1 glycosites 1500:1509 five N- and one O-linked glycosites 1475:1509 five N- and one O-linked glycosites 1475:1509 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 7 69 with glycopeptides 982:994 arg1 backbone 1022:1029 an identical peptide backbone 1001:1029 an identical peptide backbone 1001:1029 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 10 70 theme hybrid/complex 1645:1658 arg1 glycans 1660:1666 hybrid/complex glycans 1645:1666 hybrid/complex glycans 1645:1666 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 7 71 theme attached 1080:1087 arg1 structures 1096:1105 the attached glycan structures 1076:1105 the attached glycan structures 1076:1105 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 8 72 theme mass 1284:1287 arg1 correction 1289:1298 precursor mass correction 1274:1298 precursor mass correction 1274:1298 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 9 73 gly glycosites 1500:1509 arg2 glycosites 1500:1509 five N- and one O-linked glycosites 1475:1509 five N- and one O-linked glycosites 1475:1509 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 7 74 theme glycan 1089:1094 arg1 structures 1096:1105 the attached glycan structures 1076:1105 the attached glycan structures 1076:1105 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 0 75 theme protein 76:82 arg1 gp120 84:88 envelope protein gp120 67:88 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 1 76 theme Envelope 165:172 arg1 gp120 182:186 Envelope protein gp120 165:186 Envelope protein gp120 of human immunodeficiency virus (HIV) 165:224 Envelope protein gp120 of human immunodeficiency virus (HIV) is armored with a dense glycan shield, which plays critical roles in envelope folding, immune-evasion, infectivity, and immunogenicity. 24941220 6 77 gly glycosylation 957:969 arg2 glycosylation 957:969 glycosylation 957:969 glycosylation 957:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 6 77 gly glycosylation 957:969 arg2 site 949:952 the site 945:952 the site of glycosylation 945:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 11 78 theme glycan 1763:1768 arg1 Gal1GalNAc1 1770:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 6 79 gly glycopeptides 806:818 arg2 glycopeptides 806:818 glycopeptides 806:818 glycopeptides 806:818 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 11 80 theme O-linked 1799:1806 arg1 glycosite 1808:1816 the O-linked glycosite 1795:1816 the O-linked glycosite 1795:1816 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 0 81 theme energy 101:106 arg1 dissociation 120:131 higher energy collisional dissociation and spectral-aligning strategy 94:162 dissociation 120:131 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 9 82 link O-linked 1491:1498 arg1 glycosites 1500:1509 five N- and one O-linked glycosites 1475:1509 five N- and one O-linked glycosites 1475:1509 Strikingly, we were able to observe five N- and one O-linked glycosites in native gp120. 24941220 8 83 theme recombinant 1116:1126 arg1 gp120 1128:1132 the recombinant gp120 1112:1132 the recombinant gp120 1112:1132 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 7 84 theme identical 1004:1012 arg1 backbone 1022:1029 an identical peptide backbone 1001:1029 an identical peptide backbone 1001:1029 Moreover, glycopeptides with an identical peptide backbone show nearly resembling spectra regardless of the attached glycan structures. 24941220 11 85 link O-linked 1754:1761 arg1 Gal1GalNAc1 1770:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 1747:1780 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 8 86 theme O-linked 1396:1403 arg1 glycosites 1426:1435 glycosites 1426:1435 glycosites 1426:1435 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 86 theme O-linked 1396:1403 arg1 sites 1419:1423 one O-linked glycosylation sites 1392:1423 one O-linked glycosylation sites (glycosites) 1392:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 2 87 theme gp120 415:419 arg1 profiling 390:398 Site-specific glycosylation profiling 362:398 Site-specific glycosylation profiling of recombinant gp120 362:419 Site-specific glycosylation profiling of recombinant gp120 is very challenging. 24941220 12 88 theme vaccine 1949:1955 arg1 design 1957:1962 vaccine design 1949:1962 vaccine design 1949:1962 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 4 89 theme mass 578:581 arg1 spectrometer 583:594 a Q-Exactive mass spectrometer 565:594 a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120 565:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 3 90 theme glycoproteomic 453:466 arg1 analysis 468:475 glycoproteomic analysis 453:475 glycoproteomic analysis of native viral gp120 453:497 Therefore, glycoproteomic analysis of native viral gp120 is still formidable to date. 24941220 0 91 theme recombinant 22:32 arg1 virus 61:65 recombinant and human immunodeficiency virus 22:65 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 4 92 gly glycopeptides 621:633 arg1 gp120 658:662 virion-associated gp120 640:662 virion-associated gp120 640:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 4 92 gly glycopeptides 621:633 arg2 glycopeptides 621:633 low abundant glycopeptides 608:633 low abundant glycopeptides from virion-associated gp120 608:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 0 93 theme human 38:42 arg1 virus 61:65 recombinant and human immunodeficiency virus 22:65 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 8 94 gly glycosylation 1405:1417 arg2 glycosites 1426:1435 glycosites 1426:1435 glycosites 1426:1435 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 94 gly glycosylation 1405:1417 arg2 sites 1419:1423 one O-linked glycosylation sites 1392:1423 one O-linked glycosylation sites (glycosites) 1392:1436 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 94 gly glycosylation 1405:1417 arg2 one 1392:1394 one 1392:1394 one 1392:1394 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 8 95 theme " 1151:1151 arg1 pattern 1162:1168 this "copy-paste" spectral pattern 1135:1168 this "copy-paste" spectral pattern of glycopeptides 1135:1185 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 6 96 theme y-ions 918:923 arg1 terms 902:906 terms 902:906 terms of b- and y-ions that do not contain the site of glycosylation 902:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 0 97 dep virus 61:65 arg1 gp120 84:88 envelope protein gp120 67:88 recombinant and human immunodeficiency virus envelope protein gp120 22:88 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 3 98 theme viral 487:491 arg1 gp120 493:497 native viral gp120 480:497 native viral gp120 480:497 Therefore, glycoproteomic analysis of native viral gp120 is still formidable to date. 24941220 0 99 theme virus 61:65 arg1 analysis 10:17 Glycoform analysis 0:17 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 0:163 Glycoform analysis of recombinant and human immunodeficiency virus envelope protein gp120 via higher energy collisional dissociation and spectral-aligning strategy. 24941220 12 100 theme glycosylation 1870:1882 arg1 observation 1841:1851 This direct observation 1829:1851 This direct observation of site-specific glycosylation of virion-derived gp120 1829:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 24941220 6 101 theme b- 911:912 arg1 terms 902:906 terms 902:906 terms of b- and y-ions that do not contain the site of glycosylation 902:969 This strategy depends on the observation that glycopeptides and the corresponding deglycosylated peptides share very similar MS/MS pattern in terms of b- and y-ions that do not contain the site of glycosylation. 24941220 10 102 contain contain 1615:1621 arg2 glycans 1660:1666 hybrid/complex glycans 1645:1666 hybrid/complex glycans 1645:1666 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 10 102 contain contain 1615:1621 arg2 mannose 1633:1639 high mannose 1628:1639 high mannose 1628:1639 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 10 102 contain contain 1615:1621 arg1 glycans 1589:1595 glycans 1589:1595 glycans 1589:1595 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 10 103 link N-linked 1688:1695 arg1 glycosites 1697:1706 an additional four N-linked glycosites 1669:1706 an additional four N-linked glycosites 1669:1706 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 10 104 theme high 1728:1731 arg1 type 1741:1744 high mannose type 1728:1744 high mannose type 1728:1744 We further revealed that except for Asn276 in the C2 region, glycans were processed to contain both high mannose and hybrid/complex glycans; an additional four N-linked glycosites were decorated with high mannose type. 24941220 8 105 theme glycopeptides 1173:1185 arg1 pattern 1162:1168 this "copy-paste" spectral pattern 1135:1168 this "copy-paste" spectral pattern of glycopeptides 1135:1185 For the recombinant gp120, this "copy-paste" spectral pattern of glycopeptides facilitated identification of 2224 spectra using only 18 spectral templates, and after precursor mass correction, 1268 (57%) spectra were assigned to 460 unique glycopeptides accommodating 19 N-linked and one O-linked glycosylation sites (glycosites). 24941220 4 106 theme virion-associated 640:656 arg1 gp120 658:662 virion-associated gp120 640:662 virion-associated gp120 640:662 This challenge promoted us to employ a Q-Exactive mass spectrometer to identify low abundant glycopeptides from virion-associated gp120. 24941220 11 107 gly glycosite 1808:1816 arg2 glycosite 1808:1816 the O-linked glycosite 1795:1816 the O-linked glycosite 1795:1816 Core 1 O-linked glycan Gal1GalNAc1 was seen for the O-linked glycosite at Thr499. 24941220 2 108 theme glycosylation 376:388 arg1 profiling 390:398 Site-specific glycosylation profiling 362:398 Site-specific glycosylation profiling of recombinant gp120 362:419 Site-specific glycosylation profiling of recombinant gp120 is very challenging. 24941220 12 109 link virion-derived 1887:1900 arg1 gp120 1902:1906 virion-derived gp120 1887:1906 virion-derived gp120 1887:1906 This direct observation of site-specific glycosylation of virion-derived gp120 has implications in HIV glycobiology and vaccine design. 28336547 10 0 theme glycans 2184:2190 arg1 replacement 2165:2175 replacement 2165:2175 replacement of N50 glycans with structurally dissimilar biotin derivatives 2165:2238 This effect could be largely reversed by replacement of N50 glycans with structurally dissimilar biotin derivatives. 28336547 8 1 theme proteinase 1641:1650 arg1 K 1652:1652 proteinase K 1641:1652 proteinase K 1641:1652 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 2 link N-linked 1736:1743 arg1 glycans 1745:1751 N-linked glycans 1736:1751 N-linked glycans 1736:1751 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 7 3 theme biotin 1369:1374 arg1 derivatives 1376:1386 biotin derivatives 1369:1386 biotin derivatives 1369:1386 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 8 4 theme N50-linked 1818:1827 arg1 glycans 1829:1835 N50-linked glycans 1818:1835 N50-linked glycans 1818:1835 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 3 5 theme site 507:510 arg1 N50 512:514 the individual N-glycosylation site N50 476:514 the individual N-glycosylation site N50 476:514 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 7 6 theme comparable 1389:1398 arg1 derivatives 1376:1386 biotin derivatives 1369:1386 biotin derivatives 1369:1386 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 6 7 theme flux 1198:1201 arg1 studies 1203:1209 tracer flux studies 1191:1209 tracer flux studies 1191:1209 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 8 8 theme intrinsic 1676:1684 arg1 NOTEWORTHY 1773:1782 detected.NEW & NOTEWORTHY 1758:1782 detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1758:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 8 theme intrinsic 1676:1684 arg1 stability 1686:1694 a remarkable intrinsic stability 1663:1694 a remarkable intrinsic stability 1663:1694 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 6 9 theme PEPT1 1284:1288 arg1 N50Q 1290:1293 PEPT1 N50Q 1284:1293 PEPT1 N50Q 1284:1293 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 5 10 theme colonic 1001:1007 arg1 cells 1028:1032 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 cells 1028:1032 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 2 11 theme proton-coupled 291:304 arg1 transporter 319:329 the proton-coupled oligopeptide transporter 1 287:331 the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine 287:356 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 11 theme proton-coupled 291:304 arg1 PEPT1 334:338 PEPT1 334:338 PEPT1 334:338 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 6 12 theme tracer 1191:1196 arg1 studies 1203:1209 tracer flux studies 1191:1209 tracer flux studies 1191:1209 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 8 13 theme transport 1869:1877 arg1 activity 1879:1886 the bidirectional transport activity 1851:1886 the bidirectional transport activity of the murine peptide transporter PEPT1 1851:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 4 14 theme PEPT1 838:842 arg1 kinetics 854:861 PEPT1 transport kinetics 838:861 PEPT1 transport kinetics 838:861 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 6 15 theme transport 1252:1260 arg1 velocity 1262:1269 transport velocity 1252:1269 transport velocity 1252:1269 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 9 16 theme Electrophysiological 1929:1948 arg1 measurements 1966:1977 Electrophysiological and tracer flux measurements 1929:1977 measurements 1966:1977 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 8 17 theme murine 1895:1900 arg1 PEPT1 1922:1926 the murine peptide transporter PEPT1 1891:1926 the murine peptide transporter PEPT1 1891:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 1 18 theme many 127:130 arg1 proteins 141:148 many membrane proteins 127:148 many membrane proteins 127:148 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 9 19 theme tracer 1954:1959 arg1 measurements 1966:1977 Electrophysiological and tracer flux measurements 1929:1977 measurements 1966:1977 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 8 20 theme transporter 1910:1920 arg1 PEPT1 1922:1926 the murine peptide transporter PEPT1 1891:1926 the murine peptide transporter PEPT1 1891:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 21 theme N-linked 1736:1743 arg1 glycans 1745:1751 N-linked glycans 1736:1751 N-linked glycans 1736:1751 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 5 22 theme transporter 1062:1072 arg1 N50Q 1074:1077 the PEPT1 transporter N50Q 1052:1077 the PEPT1 transporter N50Q 1052:1077 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 1 23 theme extracellular 156:168 arg1 glycans 170:176 extracellular glycans 156:176 extracellular glycans 156:176 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 0 24 from Glycans 0:6 arg1 PEPT1 46:50 the intestinal peptide transporter PEPT1 11:50 the intestinal peptide transporter PEPT1 11:50 Glycans in the intestinal peptide transporter PEPT1 contribute to function and protect from proteolysis. 28336547 9 25 theme Xenopus 1982:1988 arg1 oocytes 1990:1996 Xenopus oocytes 1982:1996 Xenopus oocytes 1982:1996 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 4 26 theme sequon 722:727 arg1 N50 729:731 sequon N50 722:731 sequon N50 722:731 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 8 27 theme detected.NEW 1758:1769 arg1 NOTEWORTHY 1773:1782 detected.NEW & NOTEWORTHY 1758:1782 detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1758:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 27 theme detected.NEW 1758:1769 arg1 stability 1686:1694 a remarkable intrinsic stability 1663:1694 a remarkable intrinsic stability 1663:1694 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 4 28 gly glycosylation 705:717 arg2 N50 729:731 sequon N50 722:731 sequon N50 722:731 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 28 gly glycosylation 705:717 arg1 N50 729:731 sequon N50 722:731 sequon N50 722:731 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 9 29 from measurements 1966:1977 arg1 oocytes 1990:1996 Xenopus oocytes 1982:1996 Xenopus oocytes 1982:1996 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 5 30 theme wild-type 1127:1135 arg1 protein 1137:1143 the wild-type protein 1123:1143 the wild-type protein 1123:1143 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 10 31 theme biotin 2221:2226 arg1 derivatives 2228:2238 structurally dissimilar biotin derivatives 2197:2238 structurally dissimilar biotin derivatives 2197:2238 This effect could be largely reversed by replacement of N50 glycans with structurally dissimilar biotin derivatives. 28336547 9 32 theme outward 2104:2110 arg1 directions 2112:2121 the inward and outward directions 2089:2121 the inward and outward directions 2089:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 4 33 from inhibition 691:700 arg1 N50 729:731 sequon N50 722:731 sequon N50 722:731 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 34 theme glycosylation 705:717 arg1 inhibition 691:700 the inhibition 687:700 the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine 687:820 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 9 35 theme N50 2029:2031 arg1 glycans 2033:2039 the N50 glycans 2025:2039 the N50 glycans 2025:2039 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 2 36 dep N50 402:404 arg1 N515 425:428 N515 425:428 N515 425:428 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 36 dep N50 402:404 arg1 N532 435:438 N532 435:438 N532 435:438 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 36 dep N50 402:404 arg1 N510 419:422 N510 419:422 N510 419:422 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 36 dep N50 402:404 arg1 N439 413:416 N439 413:416 N439 413:416 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 0 37 theme peptide 26:32 arg1 PEPT1 46:50 the intestinal peptide transporter PEPT1 11:50 the intestinal peptide transporter PEPT1 11:50 Glycans in the intestinal peptide transporter PEPT1 contribute to function and protect from proteolysis. 28336547 7 38 theme PEPT1 1451:1455 arg1 transporter 1462:1472 the PEPT1 N50C transporter 1447:1472 the PEPT1 N50C transporter 1447:1472 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 5 39 theme murine 965:970 arg1 MODE-K 990:995 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 MODE-K 990:995 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 3 40 theme PEPT1-mediated 600:613 arg1 transport 622:630 the PEPT1-mediated inward transport 596:630 the PEPT1-mediated inward transport of peptides 596:642 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 5 41 theme intestinal 978:987 arg1 MODE-K 990:995 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 MODE-K 990:995 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 9 42 theme maximal 2055:2061 arg1 rate 2081:2084 the maximal peptide transport rate 2051:2084 the maximal peptide transport rate in the inward and outward directions 2051:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 8 43 theme PEPT1 1584:1588 arg1 glycosylation 1567:1579 glycosylation 1567:1579 glycosylation of PEPT1 1567:1588 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 1 44 theme glycan 213:218 arg1 chains 220:225 the glycan chains 209:225 the glycan chains 209:225 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 3 45 theme peptides 635:642 arg1 transport 622:630 the PEPT1-mediated inward transport 596:630 the PEPT1-mediated inward transport of peptides 596:642 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 2 46 dep residues 392:399 arg1 N50 402:404 N50 402:404 N50 402:404 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 7 47 with weight 1403:1408 arg1 monosaccharides 1427:1441 two to four monosaccharides 1415:1441 two to four monosaccharides 1415:1441 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 10 48 with replacement 2165:2175 arg1 derivatives 2228:2238 structurally dissimilar biotin derivatives 2197:2238 structurally dissimilar biotin derivatives 2197:2238 This effect could be largely reversed by replacement of N50 glycans with structurally dissimilar biotin derivatives. 28336547 8 49 theme proteolytic 1617:1627 arg1 cleavage 1629:1636 proteolytic cleavage 1617:1636 proteolytic cleavage by proteinase K 1617:1652 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 1 50 theme protein 239:245 arg1 function 247:254 protein function 239:254 protein function 239:254 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 9 51 theme inward 2093:2098 arg1 directions 2112:2121 the inward and outward directions 2089:2121 the inward and outward directions 2089:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 3 52 theme N50 512:514 arg1 disruption 462:471 Mutagenesis-induced disruption 442:471 Mutagenesis-induced disruption 442:471 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 7 53 theme derivatives 1376:1386 arg1 attachment 1355:1364 attachment 1355:1364 attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter 1355:1472 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 1 54 contain carry 150:154 arg1 proteins 141:148 many membrane proteins 127:148 many membrane proteins 127:148 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 1 54 contain carry 150:154 arg2 glycans 170:176 extracellular glycans 156:176 extracellular glycans 156:176 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 8 55 theme glycans 1829:1835 arg1 role 1810:1813 the role 1806:1813 the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1806:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 3 56 theme N-glycosylation 491:505 arg1 N50 512:514 the individual N-glycosylation site N50 476:514 the individual N-glycosylation site N50 476:514 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 8 57 dep NOTEWORTHY 1773:1782 arg1 highlights 1795:1804 highlights 1795:1804 highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1795:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 7 58 from comparable 1389:1398 arg1 weight 1403:1408 weight 1403:1408 weight with two to four monosaccharides 1403:1441 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 8 59 theme bidirectional 1855:1867 arg1 activity 1879:1886 the bidirectional transport activity 1851:1886 the bidirectional transport activity of the murine peptide transporter PEPT1 1851:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 2 60 theme oligopeptide 306:317 arg1 transporter 319:329 the proton-coupled oligopeptide transporter 1 287:331 the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine 287:356 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 60 theme oligopeptide 306:317 arg1 PEPT1 334:338 PEPT1 334:338 PEPT1 334:338 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 8 61 theme remarkable 1665:1674 arg1 NOTEWORTHY 1773:1782 detected.NEW & NOTEWORTHY 1758:1782 detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1758:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 61 theme remarkable 1665:1674 arg1 stability 1686:1694 a remarkable intrinsic stability 1663:1694 a remarkable intrinsic stability 1663:1694 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 7 62 from weight 1403:1408 arg1 comparable 1389:1398 comparable 1389:1398 comparable 1389:1398 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 5 63 theme epithelial 1009:1018 arg1 cells 1028:1032 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 cells 1028:1032 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 5 64 theme PTK-6 1021:1025 arg1 cells 1028:1032 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 cells 1028:1032 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 6 65 theme electrophysiological 1155:1174 arg1 recordings 1176:1185 electrophysiological recordings 1155:1185 electrophysiological recordings 1155:1185 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 7 66 attach attachment 1355:1364 arg2 derivatives 1376:1386 biotin derivatives 1369:1386 biotin derivatives 1369:1386 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 7 66 attach attachment 1355:1364 arg1 transporter 1462:1472 the PEPT1 N50C transporter 1447:1472 the PEPT1 N50C transporter 1447:1472 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 8 67 theme peptide 1902:1908 arg1 PEPT1 1922:1926 the murine peptide transporter PEPT1 1891:1926 the murine peptide transporter PEPT1 1891:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 68 theme glycans 1745:1751 arg1 absence 1725:1731 the absence 1721:1731 the absence of N-linked glycans 1721:1751 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 4 69 dep inhibition 691:700 arg1 substituting 736:747 substituting 736:747 substituting N50 with glutamine, lysine, or cysteine 736:787 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 69 dep inhibition 691:700 arg1 replacing 795:803 replacing 795:803 replacing S52 with alanine 795:820 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 70 theme transport 844:852 arg1 kinetics 854:861 PEPT1 transport kinetics 838:861 PEPT1 transport kinetics 838:861 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 71 theme murine 672:677 arg1 protein 679:685 the murine protein 668:685 the murine protein 668:685 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 8 72 theme PEPT1 1922:1926 arg1 activity 1879:1886 the bidirectional transport activity 1851:1886 the bidirectional transport activity of the murine peptide transporter PEPT1 1851:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 73 link N50-linked 1818:1827 arg1 glycans 1829:1835 N50-linked glycans 1818:1835 N50-linked glycans 1818:1835 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 1 74 theme membrane 132:139 arg1 proteins 141:148 many membrane proteins 127:148 many membrane proteins 127:148 Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. 28336547 9 75 theme flux 1961:1964 arg1 measurements 1966:1977 Electrophysiological and tracer flux measurements 1929:1977 measurements 1966:1977 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 5 76 theme PEPT1 1056:1060 arg1 N50Q 1074:1077 the PEPT1 transporter N50Q 1052:1077 the PEPT1 transporter N50Q 1052:1077 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 7 77 theme transport 1489:1497 arg1 velocity 1499:1506 the transport velocity 1485:1506 the transport velocity 1485:1506 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 3 78 gly N-glycosylation 491:505 arg2 site 507:510 the individual N-glycosylation site N50 476:514 the individual N-glycosylation site N50 476:514 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 3 78 gly N-glycosylation 491:505 arg2 N50 512:514 the individual N-glycosylation site N50 476:514 the individual N-glycosylation site N50 476:514 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 7 79 with line 1316:1319 arg1 findings 1332:1339 these findings 1326:1339 these findings 1326:1339 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 4 80 from N50 729:731 arg1 glycosylation 705:717 glycosylation 705:717 glycosylation at sequon N50 705:731 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 4 80 from N50 729:731 arg1 inhibition 691:700 the inhibition 687:700 the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine 687:820 Here, we show that for the murine protein the inhibition of glycosylation at sequon N50 by substituting N50 with glutamine, lysine, or cysteine or by replacing S52 with alanine equally altered PEPT1 transport kinetics in oocytes. 28336547 2 81 from transporter 319:329 arg1 intestine 348:356 the intestine 344:356 the intestine 344:356 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 10 82 theme N50 2180:2182 arg1 glycans 2184:2190 N50 glycans 2180:2190 N50 glycans 2180:2190 This effect could be largely reversed by replacement of N50 glycans with structurally dissimilar biotin derivatives. 28336547 2 83 theme asparagine 381:390 arg1 residues 392:399 six asparagine residues 377:399 six asparagine residues (N50, N406, N439, N510, N515, and N532) 377:439 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 8 84 theme & 1771:1771 arg1 NOTEWORTHY 1773:1782 detected.NEW & NOTEWORTHY 1758:1782 detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1758:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 84 theme & 1771:1771 arg1 stability 1686:1694 a remarkable intrinsic stability 1663:1694 a remarkable intrinsic stability 1663:1694 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 9 85 theme transport 2071:2079 arg1 rate 2081:2084 the maximal peptide transport rate 2051:2084 the maximal peptide transport rate in the inward and outward directions 2051:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 10 86 theme dissimilar 2210:2219 arg1 derivatives 2228:2238 structurally dissimilar biotin derivatives 2197:2238 structurally dissimilar biotin derivatives 2197:2238 This effect could be largely reversed by replacement of N50 glycans with structurally dissimilar biotin derivatives. 28336547 11 87 located detected 2269:2276 arg1 addition 2244:2251 addition 2244:2251 addition 2244:2251 In addition, N-glycans were detected to stabilize PEPT1 against proteolytic cleavage. 28336547 11 87 located detected 2269:2276 arg2 N-glycans 2254:2262 N-glycans 2254:2262 N-glycans 2254:2262 In addition, N-glycans were detected to stabilize PEPT1 against proteolytic cleavage. 28336547 0 88 theme intestinal 15:24 arg1 PEPT1 46:50 the intestinal peptide transporter PEPT1 11:50 the intestinal peptide transporter PEPT1 11:50 Glycans in the intestinal peptide transporter PEPT1 contribute to function and protect from proteolysis. 28336547 5 89 theme glycyl-sarcosine 932:947 arg1 uptake 917:922 the uptake 913:922 the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q 913:1077 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 2 90 gly glycosylated 361:372 arg2 residues 392:399 six asparagine residues 377:399 six asparagine residues (N50, N406, N439, N510, N515, and N532) 377:439 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 90 gly glycosylated 361:372 arg1 transporter 319:329 the proton-coupled oligopeptide transporter 1 287:331 the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine 287:356 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 2 90 gly glycosylated 361:372 arg1 PEPT1 334:338 PEPT1 334:338 PEPT1 334:338 We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, and N532). 28336547 3 91 theme inward 615:620 arg1 transport 622:630 the PEPT1-mediated inward transport 596:630 the PEPT1-mediated inward transport of peptides 596:642 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 0 92 theme transporter 34:44 arg1 PEPT1 46:50 the intestinal peptide transporter PEPT1 11:50 the intestinal peptide transporter PEPT1 11:50 Glycans in the intestinal peptide transporter PEPT1 contribute to function and protect from proteolysis. 28336547 7 93 dep four 1422:1425 arg1 to 1419:1420 to 1419:1420 to 1419:1420 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 5 94 theme immortalized 952:963 arg1 MODE-K 990:995 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 MODE-K 990:995 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 9 95 theme glycans 2033:2039 arg1 removal 2014:2020 removal 2014:2020 removal of the N50 glycans 2014:2039 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 9 96 from rate 2081:2084 arg1 directions 2112:2121 the inward and outward directions 2089:2121 the inward and outward directions 2089:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 6 97 from rise 1244:1247 arg1 velocity 1262:1269 transport velocity 1252:1269 transport velocity 1252:1269 By using electrophysiological recordings and tracer flux studies, we further demonstrate that the rise in transport velocity observed for PEPT1 N50Q is bidirectional. 28336547 7 98 theme N50C 1457:1460 arg1 transporter 1462:1472 the PEPT1 N50C transporter 1447:1472 the PEPT1 N50C transporter 1447:1472 In line with these findings, we show that attachment of biotin derivatives, comparable in weight with two to four monosaccharides, to the PEPT1 N50C transporter slows down the transport velocity. 28336547 5 99 theme small 972:976 arg1 MODE-K 990:995 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 MODE-K 990:995 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 3 100 theme Mutagenesis-induced 442:460 arg1 disruption 462:471 Mutagenesis-induced disruption 442:471 Mutagenesis-induced disruption 442:471 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 11 101 theme proteolytic 2305:2315 arg1 cleavage 2317:2324 proteolytic cleavage 2305:2324 proteolytic cleavage 2305:2324 In addition, N-glycans were detected to stabilize PEPT1 against proteolytic cleavage. 28336547 9 102 theme peptide 2063:2069 arg1 rate 2081:2084 the maximal peptide transport rate 2051:2084 the maximal peptide transport rate in the inward and outward directions 2051:2121 Electrophysiological and tracer flux measurements in Xenopus oocytes have shown that removal of the N50 glycans increases the maximal peptide transport rate in the inward and outward directions. 28336547 8 103 from stability 1686:1694 arg1 absence 1725:1731 the absence 1721:1731 the absence of N-linked glycans 1721:1751 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 104 gly glycosylation 1567:1579 arg1 PEPT1 1584:1588 PEPT1 1584:1588 PEPT1 1584:1588 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 8 105 theme strong 1546:1551 arg1 evidence 1553:1560 strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1546:1926 strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1 1546:1926 In addition, our experiments provide strong evidence that glycosylation of PEPT1 confers resistance against proteolytic cleavage by proteinase K, whereas a remarkable intrinsic stability against trypsin, even in the absence of N-linked glycans, was detected.NEW & NOTEWORTHY This study highlights the role of N50-linked glycans in modulating the bidirectional transport activity of the murine peptide transporter PEPT1. 28336547 3 106 theme individual 480:489 arg1 N50 512:514 the individual N-glycosylation site N50 476:514 the individual N-glycosylation site N50 476:514 Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1-mediated inward transport of peptides. 28336547 5 107 from uptake 917:922 arg1 MODE-K 990:995 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 MODE-K 990:995 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 28336547 5 107 from uptake 917:922 arg1 cells 1028:1032 immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells 952:1032 cells 1028:1032 Furthermore, we provide evidence that the uptake of [14C]glycyl-sarcosine in immortalized murine small intestinal (MODE-K) or colonic epithelial (PTK-6) cells stably expressing the PEPT1 transporter N50Q is also significantly increased relative to the wild-type protein. 27063138 0 0 theme Chinese 74:80 arg1 cells 102:106 Chinese Hamster Ovary (CHO) cells 74:106 Chinese Hamster Ovary (CHO) cells 74:106 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 3 1 from production 394:403 arg1 isolation 337:345 isolation 337:345 isolation from either native sources or the heterologous production in insect cells 337:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 3 1 from production 394:403 arg1 cells 415:419 insect cells 408:419 insect cells 408:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 11 2 theme previous 1304:1311 arg1 publications 1313:1324 previous publications 1304:1324 previous publications 1304:1324 The substrate preference and kinetic parameters were in accordance with previous publications. 27063138 4 3 theme non-human 488:496 arg1 patterns 512:519 non-human glycosylation patterns 488:519 non-human glycosylation patterns that may alter its biochemical properties 488:561 Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. 27063138 5 4 theme hDAO 606:609 arg1 expression 592:601 the heterologous expression 575:601 the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells 575:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 5 4 theme hDAO 606:609 arg1 protocol 678:685 a three step purification protocol 652:685 a three step purification protocol 652:685 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 7 5 theme phenylhydrazine 846:860 arg1 titration 862:870 phenylhydrazine titration 846:870 phenylhydrazine titration 846:870 Topaquinone (TPQ) cofactor content was determined using phenylhydrazine titration. 27063138 0 6 theme Ovary 90:94 arg1 cells 102:106 Chinese Hamster Ovary (CHO) cells 74:106 Chinese Hamster Ovary (CHO) cells 74:106 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 2 7 theme histamine 273:281 arg1 intolerance 283:293 histamine intolerance 273:293 histamine intolerance 273:293 Reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. 27063138 12 8 theme decent 1412:1417 arg1 protein 1430:1436 protein 1430:1436 protein 1430:1436 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 12 8 theme decent 1412:1417 arg1 amounts 1419:1425 decent amounts 1412:1425 decent amounts of protein 1412:1436 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 10 9 theme structural 1209:1218 arg1 properties 1220:1229 structural properties 1209:1229 structural properties 1209:1229 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 0 10 theme Hamster 82:88 arg1 cells 102:106 Chinese Hamster Ovary (CHO) cells 74:106 Chinese Hamster Ovary (CHO) cells 74:106 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 4 11 theme insect 467:472 arg1 cells 474:478 insect cells 467:478 insect cells 467:478 Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. 27063138 5 12 theme Chinese 614:620 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 6 13 theme content 706:712 arg1 Analysis 688:695 Analysis 688:695 Analysis of metal content using ICP-MS 688:725 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 0 14 theme CHO 97:99 arg1 cells 102:106 Chinese Hamster Ovary (CHO) cells 74:106 Chinese Hamster Ovary (CHO) cells 74:106 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 3 15 from sources 366:372 arg1 isolation 337:345 isolation 337:345 isolation from either native sources or the heterologous production in insect cells 337:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 3 15 from sources 366:372 arg1 cells 415:419 insect cells 408:419 insect cells 408:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 5 16 theme Ovary 630:634 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 5 17 theme Hamster 622:628 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 12 18 theme negligible 1443:1452 arg1 impurities 1454:1463 negligible impurities to address new scientific questions 1443:1499 negligible impurities to address new scientific questions 1443:1499 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 6 19 theme metal 700:704 arg1 content 706:712 metal content 700:712 metal content 700:712 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 10 20 used used 1188:1191 arg2 data 1178:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 8 21 theme active 964:969 arg1 site 971:974 the active site 960:974 the active site 960:974 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 8 22 theme Ninety-four 873:883 arg1 percent 885:891 Ninety-four percent 873:891 Ninety-four percent of DAO molecules 873:908 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 8 23 contain contained 910:918 arg2 TPQ 920:922 TPQ 920:922 TPQ 920:922 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 8 23 contain contained 910:918 arg1 percent 885:891 Ninety-four percent 873:891 Ninety-four percent of DAO molecules 873:908 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 5 24 theme CHO 637:639 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 5 25 theme heterologous 579:590 arg1 expression 592:601 the heterologous expression 575:601 the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells 575:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 10 26 theme circular 1140:1147 arg1 dichroism 1149:1157 Electronic circular dichroism 1129:1157 Electronic circular dichroism 1129:1157 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 6 27 theme sites 759:763 arg1 % 743:743 93% 741:743 93% of the active sites 741:763 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 6 27 theme sites 759:763 arg1 sites 759:763 the active sites 748:763 the active sites 748:763 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 6 28 used occupied 770:777 arg2 % 743:743 93% 741:743 93% of the active sites 741:763 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 6 28 used occupied 770:777 arg2 sites 759:763 the active sites 748:763 the active sites 748:763 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 3 29 theme native 359:364 arg1 sources 366:372 native sources 359:372 native sources 359:372 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 7 30 theme Topaquinone 790:800 arg1 content 817:823 Topaquinone (TPQ) cofactor content 790:823 Topaquinone (TPQ) cofactor content 790:823 Topaquinone (TPQ) cofactor content was determined using phenylhydrazine titration. 27063138 6 31 theme active 752:757 arg1 sites 759:763 the active sites 748:763 the active sites 748:763 Analysis of metal content using ICP-MS revealed that 93% of the active sites were occupied by copper. 27063138 7 32 theme cofactor 808:815 arg1 content 817:823 Topaquinone (TPQ) cofactor content 790:823 Topaquinone (TPQ) cofactor content 790:823 Topaquinone (TPQ) cofactor content was determined using phenylhydrazine titration. 27063138 0 33 theme recombinant 20:30 arg1 oxidase 46:52 recombinant human diamine oxidase 20:52 recombinant human diamine oxidase (rhDAO) 20:60 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 0 33 theme recombinant 20:30 arg1 rhDAO 55:59 rhDAO 55:59 rhDAO 55:59 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 11 34 theme substrate 1236:1244 arg1 preference 1246:1255 The substrate preference 1232:1255 The substrate preference 1232:1255 The substrate preference and kinetic parameters were in accordance with previous publications. 27063138 9 35 theme glycosylation 1106:1118 arg1 profile 1120:1126 the glycosylation profile 1102:1126 the glycosylation profile 1102:1126 Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. 27063138 8 36 theme molecules 900:908 arg1 percent 885:891 Ninety-four percent 873:891 Ninety-four percent of DAO molecules 873:908 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 0 37 theme oxidase 46:52 arg1 Characterization 0:15 Characterization 0:15 Characterization of recombinant human diamine oxidase (rhDAO) 0:60 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 4 38 theme human 439:443 arg1 enzyme 445:450 human enzyme 439:450 human enzyme 439:450 Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. 27063138 9 39 theme sequence 1054:1061 arg1 integrity 1063:1071 sequence integrity 1054:1071 sequence integrity of the protein 1054:1086 Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. 27063138 8 40 theme DAO 896:898 arg1 molecules 900:908 DAO molecules 896:908 DAO molecules 896:908 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 0 41 theme diamine 38:44 arg1 oxidase 46:52 recombinant human diamine oxidase 20:52 recombinant human diamine oxidase (rhDAO) 20:60 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 0 41 theme diamine 38:44 arg1 rhDAO 55:59 rhDAO 55:59 rhDAO 55:59 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 10 42 theme spectra 1170:1176 arg1 data 1178:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 3 43 theme insect 408:413 arg1 cells 415:419 insect cells 408:419 insect cells 408:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 0 44 theme human 32:36 arg1 oxidase 46:52 recombinant human diamine oxidase 20:52 recombinant human diamine oxidase (rhDAO) 20:60 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 0 44 theme human 32:36 arg1 rhDAO 55:59 rhDAO 55:59 rhDAO 55:59 Characterization of recombinant human diamine oxidase (rhDAO) produced in Chinese Hamster Ovary (CHO) cells. 27063138 3 45 theme heterologous 381:392 arg1 production 394:403 the heterologous production 377:403 the heterologous production in insect cells 377:419 So far hDAO has been characterized after isolation from either native sources or the heterologous production in insect cells. 27063138 10 46 theme UV-vis 1163:1168 arg1 spectra 1170:1176 UV-vis spectra 1163:1176 UV-vis spectra 1163:1176 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 5 47 theme step 660:663 arg1 protocol 678:685 a three step purification protocol 652:685 a three step purification protocol 652:685 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 10 48 theme Electronic 1129:1138 arg1 dichroism 1149:1157 Electronic circular dichroism 1129:1157 Electronic circular dichroism 1129:1157 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 12 49 theme new 1476:1478 arg1 questions 1491:1499 new scientific questions 1476:1499 new scientific questions 1476:1499 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 5 50 from expression 592:601 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 9 51 theme Mass 1002:1005 arg1 analysis 1021:1028 Mass spectrometric analysis 1002:1028 Mass spectrometric analysis 1002:1028 Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. 27063138 4 52 theme biochemical 540:550 arg1 properties 552:561 its biochemical properties 536:561 its biochemical properties 536:561 Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. 27063138 5 53 theme purification 665:676 arg1 protocol 678:685 a three step purification protocol 652:685 a three step purification protocol 652:685 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 2 54 theme Reduced 185:191 arg1 activities 200:209 Reduced enzyme activities 185:209 Reduced enzyme activities 185:209 Reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. 27063138 2 55 theme enzyme 193:198 arg1 activities 200:209 Reduced enzyme activities 185:209 Reduced enzyme activities 185:209 Reduced enzyme activities might cause complications during pregnancy and be involved in histamine intolerance. 27063138 8 56 from site 971:974 arg1 content 949:955 the copper content 938:955 the copper content at the active site 938:974 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 9 57 theme spectrometric 1007:1019 arg1 analysis 1021:1028 Mass spectrometric analysis 1002:1028 Mass spectrometric analysis 1002:1028 Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. 27063138 10 58 theme dichroism 1149:1157 arg1 data 1178:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data 1129:1181 Electronic circular dichroism and UV-vis spectra data were used to characterize structural properties. 27063138 11 59 theme kinetic 1261:1267 arg1 parameters 1269:1278 kinetic parameters 1261:1278 kinetic parameters 1261:1278 The substrate preference and kinetic parameters were in accordance with previous publications. 27063138 12 60 theme recombinant 1350:1360 arg1 system 1373:1378 a recombinant production system 1348:1378 a recombinant production system for hDAO 1348:1387 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 5 61 from protocol 678:685 arg1 cells 642:646 Chinese Hamster Ovary (CHO) cells 614:646 Chinese Hamster Ovary (CHO) cells 614:646 We present the heterologous expression of hDAO in Chinese Hamster Ovary (CHO) cells and a three step purification protocol. 27063138 12 62 theme protein 1430:1436 arg1 protein 1430:1436 protein 1430:1436 protein 1430:1436 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 12 62 theme protein 1430:1436 arg1 amounts 1419:1425 decent amounts 1412:1425 decent amounts of protein 1412:1436 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 8 63 theme copper 942:947 arg1 content 949:955 the copper content 938:955 the copper content at the active site 938:974 Ninety-four percent of DAO molecules contained TPQ and therefore the copper content at the active site was indirectly confirmed. 27063138 1 64 theme Human 109:113 arg1 oxidase 123:129 Human diamine oxidase 109:129 Human diamine oxidase (hDAO) 109:136 Human diamine oxidase (hDAO) efficiently degrades polyamines and histamine. 27063138 1 64 theme Human 109:113 arg1 hDAO 132:135 hDAO 132:135 hDAO 132:135 Human diamine oxidase (hDAO) efficiently degrades polyamines and histamine. 27063138 9 65 theme protein 1080:1086 arg1 integrity 1063:1071 sequence integrity 1054:1071 sequence integrity of the protein 1054:1086 Mass spectrometric analysis was conducted to verify sequence integrity of the protein and to assess the glycosylation profile. 27063138 12 66 theme system 1373:1378 arg1 establishment 1331:1343 The establishment 1327:1343 The establishment of a recombinant production system for hDAO 1327:1387 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 1 67 theme diamine 115:121 arg1 oxidase 123:129 Human diamine oxidase 109:129 Human diamine oxidase (hDAO) 109:136 Human diamine oxidase (hDAO) efficiently degrades polyamines and histamine. 27063138 1 67 theme diamine 115:121 arg1 hDAO 132:135 hDAO 132:135 hDAO 132:135 Human diamine oxidase (hDAO) efficiently degrades polyamines and histamine. 27063138 4 68 theme glycosylation 498:510 arg1 patterns 512:519 non-human glycosylation patterns 488:519 non-human glycosylation patterns that may alter its biochemical properties 488:561 Accessibility to human enzyme is limited and insect cells produce non-human glycosylation patterns that may alter its biochemical properties. 27063138 12 69 theme production 1362:1371 arg1 system 1373:1378 a recombinant production system 1348:1378 a recombinant production system for hDAO 1348:1387 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 27063138 12 70 theme scientific 1480:1489 arg1 questions 1491:1499 new scientific questions 1476:1499 new scientific questions 1476:1499 The establishment of a recombinant production system for hDAO enables us to generate decent amounts of protein with negligible impurities to address new scientific questions. 25575808 0 0 theme inflammatory 78:89 arg1 reaction 91:98 stromal inflammatory reaction 70:98 stromal inflammatory reaction 70:98 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 3 1 theme mouse 326:330 arg1 proteoglycan 376:387 a chondroitin sulfate proteoglycan 354:387 a chondroitin sulfate proteoglycan 354:387 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 3 1 theme mouse 326:330 arg1 endocan 332:338 mouse endocan 326:338 mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan 326:431 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 11 2 theme tumor 1381:1385 arg1 growth 1387:1392 tumor growth 1381:1392 tumor growth 1381:1392 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 6 3 theme growth 712:717 arg1 inhibition 719:728 This tumor growth inhibition 701:728 This tumor growth inhibition 701:728 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 5 4 from overexpression 590:603 arg1 cells 631:635 HT-29 cells 625:635 HT-29 cells 625:635 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 7 5 theme anti-tumor 875:884 arg1 effect 886:891 an anti-tumor effect 872:891 an anti-tumor effect 872:891 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 0 6 theme stromal 70:76 arg1 reaction 91:98 stromal inflammatory reaction 70:98 stromal inflammatory reaction 70:98 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 5 7 theme tumor 649:653 arg1 appearance 655:664 the tumor appearance 645:664 the tumor appearance 645:664 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 10 8 theme tissue 1096:1101 arg1 sections 1103:1110 tumor tissue sections 1090:1110 tumor tissue sections 1090:1110 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 10 9 theme endocan 1188:1194 arg1 polypeptide 1196:1206 endocan polypeptide 1188:1206 endocan polypeptide 1188:1206 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 3 10 theme chondroitin 356:366 arg1 proteoglycan 376:387 a chondroitin sulfate proteoglycan 354:387 a chondroitin sulfate proteoglycan 354:387 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 3 10 theme chondroitin 356:366 arg1 endocan 332:338 mouse endocan 326:338 mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan 326:431 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 6 11 theme endocan 775:781 arg1 form 761:764 non glycanated form 746:764 non glycanated form of mouse endocan 746:781 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 8 12 theme tumor 976:980 arg1 growth 982:987 HT-29 tumor growth 970:987 HT-29 tumor growth delay 970:993 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 2 13 theme growth 289:294 arg1 processes 270:278 various processes 262:278 various processes of tumor growth 262:294 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 8 14 theme endocan 946:952 arg1 delivery 913:920 systemic delivery 904:920 systemic delivery of non glycanated human endocan 904:952 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 3 15 theme sulfate 368:374 arg1 proteoglycan 376:387 a chondroitin sulfate proteoglycan 354:387 a chondroitin sulfate proteoglycan 354:387 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 3 15 theme sulfate 368:374 arg1 endocan 332:338 mouse endocan 326:338 mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan 326:431 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 8 16 theme systemic 904:911 arg1 delivery 913:920 systemic delivery 904:920 systemic delivery of non glycanated human endocan 904:952 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 5 17 theme mouse 608:612 arg1 endocan 614:620 mouse endocan 608:620 mouse endocan 608:620 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 2 18 theme tumor 283:287 arg1 growth 289:294 tumor growth 283:294 tumor growth 283:294 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 6 19 theme tumor 706:710 arg1 inhibition 719:728 This tumor growth inhibition 701:728 This tumor growth inhibition 701:728 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 8 20 theme growth 982:987 arg1 delay 989:993 HT-29 tumor growth delay 970:993 HT-29 tumor growth delay 970:993 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 4 21 theme located 479:485 arg1 site 473:476 the O-glycanation site 455:476 the O-glycanation site 455:476 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 5 22 theme endocan 614:620 arg1 overexpression 590:603 overexpression 590:603 overexpression of mouse endocan in HT-29 cells 590:635 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 11 23 theme immunity 1443:1450 arg1 cells 1423:1427 inflammatory cells 1410:1427 inflammatory cells of the innate immunity 1410:1450 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 5 24 theme HT-29 625:629 arg1 cells 631:635 HT-29 cells 625:635 HT-29 cells 625:635 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 7 25 theme A549 837:840 arg1 cells 851:855 HT-29, A549 or K1000 cells 830:855 HT-29, A549 or K1000 cells 830:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 10 26 theme stromal 1115:1121 arg1 reaction 1136:1143 a stromal inflammatory reaction 1113:1143 a stromal inflammatory reaction 1113:1143 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 0 27 theme non 4:6 arg1 polypeptide 27:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 10 28 theme tumor 1090:1094 arg1 sections 1103:1110 tumor tissue sections 1090:1110 tumor tissue sections 1090:1110 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 5 29 theme human 573:577 arg1 homologue 579:587 the human homologue 569:587 the human homologue 569:587 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 7 30 theme human 799:803 arg1 endocan 805:811 Non glycanated human endocan 784:811 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells 784:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 6 31 theme mouse 769:773 arg1 endocan 775:781 mouse endocan 769:781 mouse endocan 769:781 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 7 32 theme HT-29 830:834 arg1 cells 851:855 HT-29, A549 or K1000 cells 830:855 HT-29, A549 or K1000 cells 830:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 0 33 theme endocan 19:25 arg1 polypeptide 27:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 8 34 theme HT-29 970:974 arg1 growth 982:987 HT-29 tumor growth 970:987 HT-29 tumor growth delay 970:993 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 1 35 theme Endocan 101:107 arg1 expression 109:118 Endocan expression 101:118 Endocan expression 101:118 Endocan expression is increasingly studied in various human cancers. 25575808 11 36 theme innate 1436:1441 arg1 immunity 1443:1450 the innate immunity 1432:1450 the innate immunity 1432:1450 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 0 37 theme glycanated 8:17 arg1 polypeptide 27:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide 0:37 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 7 38 theme Non 784:786 arg1 endocan 805:811 Non glycanated human endocan 784:811 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells 784:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 4 39 from site 473:476 arg1 domains 442:448 Distant domains 434:448 Distant domains from the O-glycanation site, located within exons 1 and 2 434:506 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 4 40 theme O-glycanation 459:471 arg1 site 473:476 the O-glycanation site 455:476 the O-glycanation site 455:476 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 9 41 theme cell 1071:1074 arg1 viability 1076:1084 cell viability 1071:1084 cell viability 1071:1084 In vitro, endocan polypeptide did not affect HT-29 cell proliferation, nor cell viability. 25575808 3 42 theme human 419:423 arg1 endocan 425:431 human endocan 419:431 human endocan 419:431 We functionally characterize mouse endocan which is also a chondroitin sulfate proteoglycan but much less glycanated than human endocan. 25575808 7 43 theme glycanated 788:797 arg1 endocan 805:811 Non glycanated human endocan 784:811 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells 784:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 10 44 theme cells 1233:1237 arg1 depletion 1213:1221 depletion 1213:1221 depletion of CD122+ cells 1213:1237 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 10 45 theme anti-tumor 1272:1281 arg1 effect 1283:1288 the anti-tumor effect 1268:1288 the anti-tumor effect of endocan polypeptide 1268:1311 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 11 46 theme inflammatory 1410:1421 arg1 cells 1423:1427 inflammatory cells 1410:1427 inflammatory cells of the innate immunity 1410:1450 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 7 47 theme K1000 845:849 arg1 cells 851:855 HT-29, A549 or K1000 cells 830:855 HT-29, A549 or K1000 cells 830:855 Non glycanated human endocan overexpressed in HT-29, A549 or K1000 cells also exhibited an anti-tumor effect. 25575808 11 48 theme novel 1337:1341 arg1 pathway 1343:1349 a novel pathway 1335:1349 a novel pathway 1335:1349 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 4 49 theme glycanation 522:532 arg1 pattern 534:540 the glycanation pattern 518:540 the glycanation pattern of endocan 518:551 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 2 50 theme Experimental 170:181 arg1 evidence 183:190 Experimental evidence 170:190 Experimental evidence 170:190 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 2 51 theme glycan 231:236 arg1 chain 238:242 its glycan chain 227:242 its glycan chain 227:242 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 6 52 theme glycanated 750:759 arg1 form 761:764 non glycanated form 746:764 non glycanated form of mouse endocan 746:781 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 0 53 theme tumor 45:49 arg1 growth 51:56 tumor growth 45:56 tumor growth 45:56 The non glycanated endocan polypeptide slows tumor growth by inducing stromal inflammatory reaction. 25575808 4 54 theme endocan 545:551 arg1 pattern 534:540 the glycanation pattern 518:540 the glycanation pattern of endocan 518:551 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 4 55 theme Distant 434:440 arg1 domains 442:448 Distant domains 434:448 Distant domains from the O-glycanation site, located within exons 1 and 2 434:506 Distant domains from the O-glycanation site, located within exons 1 and 2 determine the glycanation pattern of endocan. 25575808 8 56 theme non 925:927 arg1 endocan 946:952 non glycanated human endocan 925:952 non glycanated human endocan 925:952 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 6 57 theme non 746:748 arg1 form 761:764 non glycanated form 746:764 non glycanated form of mouse endocan 746:781 This tumor growth inhibition is supported by non glycanated form of mouse endocan. 25575808 10 58 located observed 1149:1156 arg1 sections 1103:1110 tumor tissue sections 1090:1110 tumor tissue sections 1090:1110 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 10 58 located observed 1149:1156 arg2 reaction 1136:1143 a stromal inflammatory reaction 1113:1143 a stromal inflammatory reaction 1113:1143 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 10 58 located observed 1149:1156 arg1 tumors 1166:1171 tumors 1166:1171 tumors overexpressing endocan polypeptide 1166:1206 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 5 59 theme tumor 682:686 arg1 rate 695:698 the tumor growth rate 678:698 the tumor growth rate 678:698 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 11 60 from endocan 1355:1361 arg1 control 1370:1376 the control 1366:1376 the control of tumor growth 1366:1392 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 2 61 theme various 262:268 arg1 processes 270:278 various processes 262:278 various processes of tumor growth 262:294 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 10 62 theme polypeptide 1301:1311 arg1 effect 1283:1288 the anti-tumor effect 1268:1288 the anti-tumor effect of endocan polypeptide 1268:1311 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 10 63 theme endocan 1293:1299 arg1 polypeptide 1301:1311 endocan polypeptide 1293:1311 endocan polypeptide 1293:1311 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 11 64 theme growth 1387:1392 arg1 control 1370:1376 the control 1366:1376 the control of tumor growth 1366:1392 These results reveal a novel pathway for endocan in the control of tumor growth, which involves inflammatory cells of the innate immunity. 25575808 5 65 theme growth 688:693 arg1 rate 695:698 the tumor growth rate 678:698 the tumor growth rate 678:698 In opposite to the human homologue, overexpression of mouse endocan in HT-29 cells delayed the tumor appearance and reduced the tumor growth rate. 25575808 10 66 theme CD122+ 1226:1231 arg1 cells 1233:1237 CD122+ cells 1226:1237 CD122+ cells 1226:1237 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 25575808 8 67 theme human 940:944 arg1 endocan 946:952 non glycanated human endocan 925:952 non glycanated human endocan 925:952 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 9 68 theme endocan 1006:1012 arg1 polypeptide 1014:1024 endocan polypeptide 1006:1024 endocan polypeptide 1006:1024 In vitro, endocan polypeptide did not affect HT-29 cell proliferation, nor cell viability. 25575808 2 69 theme human 204:208 arg1 endocan 210:216 human endocan 204:216 human endocan 204:216 Experimental evidence showed that human endocan, through its glycan chain, is implicated in various processes of tumor growth. 25575808 8 70 theme glycanated 929:938 arg1 endocan 946:952 non glycanated human endocan 925:952 non glycanated human endocan 925:952 Moreover, systemic delivery of non glycanated human endocan also results in HT-29 tumor growth delay. 25575808 1 71 theme various 147:153 arg1 cancers 161:167 various human cancers 147:167 various human cancers 147:167 Endocan expression is increasingly studied in various human cancers. 25575808 9 72 theme HT-29 1041:1045 arg1 proliferation 1052:1064 HT-29 cell proliferation 1041:1064 HT-29 cell proliferation 1041:1064 In vitro, endocan polypeptide did not affect HT-29 cell proliferation, nor cell viability. 25575808 1 73 theme human 155:159 arg1 cancers 161:167 various human cancers 147:167 various human cancers 147:167 Endocan expression is increasingly studied in various human cancers. 25575808 9 74 theme cell 1047:1050 arg1 proliferation 1052:1064 HT-29 cell proliferation 1041:1064 HT-29 cell proliferation 1041:1064 In vitro, endocan polypeptide did not affect HT-29 cell proliferation, nor cell viability. 25575808 10 75 theme inflammatory 1123:1134 arg1 reaction 1136:1143 a stromal inflammatory reaction 1113:1143 a stromal inflammatory reaction 1113:1143 In tumor tissue sections, a stromal inflammatory reaction was observed only in tumors overexpressing endocan polypeptide, and depletion of CD122+ cells was able to delete partially the anti-tumor effect of endocan polypeptide. 24798328 6 0 theme residue 959:965 arg1 position 925:932 position -1 925:935 position -1 of the first cysteine residue of most repeats 925:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 1 theme A 855:855 arg1 repeats 857:863 LDLR class A repeats 844:863 LDLR class A repeats 844:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 2 theme first 944:948 arg1 residue 959:965 the first cysteine residue 940:965 the first cysteine residue of most repeats 940:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 2 theme first 944:948 arg1 repeats 975:981 most repeats 970:981 most repeats 970:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 4 3 theme SimpleCell 611:620 arg1 strategy 647:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 6 4 from position 925:932 arg1 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 5 5 theme O-glycosylation 702:716 arg1 sites 718:722 O-glycosylation sites 702:722 O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells 702:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 6 6 theme LDLR 844:847 arg1 A 855:855 LDLR class A 844:855 LDLR class A repeats 844:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 2 7 from surface 334:340 arg1 expression 311:320 stable expression 304:320 stable expression at the cell surface 304:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 7 8 theme repeats 1124:1130 arg1 regions 1100:1106 linker regions 1093:1106 linker regions of LDLR class A repeats 1093:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 7 9 theme other 1187:1191 arg1 receptors 1204:1212 other homologous receptors 1187:1212 other homologous receptors 1187:1212 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 8 10 theme polypeptide 1266:1276 arg1 transferases 1285:1296 polypeptide GalNAc transferases 1266:1296 polypeptide GalNAc transferases 1266:1296 O-Glycosylation is controlled by a large family of polypeptide GalNAc transferases. 24798328 7 11 theme class 1116:1120 arg1 A 1122:1122 LDLR class A 1111:1122 LDLR class A repeats 1111:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 6 12 theme short 815:819 arg1 regions 828:834 the short linker regions 811:834 the short linker regions between LDLR class A repeats 811:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 13 theme CHO 1003:1005 arg1 cells 1007:1011 wild-type CHO cells 993:1011 wild-type CHO cells 993:1011 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 8 14 theme transferases 1285:1296 arg1 family 1256:1261 a large family 1248:1261 a large family of polypeptide GalNAc transferases 1248:1296 O-Glycosylation is controlled by a large family of polypeptide GalNAc transferases. 24798328 1 15 theme LDLR 188:191 arg1 functions 193:201 LDLR functions 188:201 LDLR functions 188:201 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 7 16 theme linker 1093:1098 arg1 regions 1100:1106 linker regions 1093:1106 linker regions of LDLR class A repeats 1093:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 0 17 from O-glycosylated 53:66 arg1 regions 78:84 linker regions 71:84 linker regions 71:84 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 7 18 gly glycosites 1079:1088 arg2 glycosites 1079:1088 The glycosites 1075:1088 The glycosites in linker regions of LDLR class A repeats 1075:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 6 19 theme most 970:973 arg1 repeats 975:981 most repeats 970:981 most repeats 970:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 2 20 theme I 246:246 arg1 LDLR 231:234 LDLR 231:234 LDLR 231:234 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 2 20 theme I 246:246 arg1 protein 262:268 a type I transmembrane protein 239:268 a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface 239:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 6 21 theme core 1057:1060 arg1 structure 1064:1072 the typical sialylated core 1 structure 1034:1072 the typical sialylated core 1 structure 1034:1072 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 22 theme typical 1038:1044 arg1 structure 1064:1072 the typical sialylated core 1 structure 1034:1072 the typical sialylated core 1 structure 1034:1072 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 7 23 from man 1159:1161 arg1 LDLR 1149:1152 LDLR 1149:1152 LDLR from man to Xenopus 1149:1172 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 0 24 theme linker 71:76 arg1 regions 78:84 linker regions 71:84 linker regions 71:84 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 9 25 theme isoform 1318:1324 arg1 s 1326:1326 isoform(s) 1318:1327 isoform(s) 1318:1327 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 5 26 theme CHO 777:779 arg1 cells 791:795 CHO wild-type cells 777:795 CHO wild-type cells 777:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 10 27 gly glycosylation 1596:1608 arg1 regions 1634:1640 three of four linker regions 1613:1640 three of four linker regions 1613:1640 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 10 28 theme regions 1634:1640 arg1 glycosylation 1596:1608 glycosylation 1596:1608 glycosylation of three of four linker regions 1596:1640 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 0 29 theme A 39:39 arg1 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 3 30 theme LDLR 381:384 arg1 O-glycosylation 386:400 LDLR O-glycosylation 381:400 LDLR O-glycosylation 381:400 It has previously been suggested that LDLR O-glycosylation is found N-terminal to the juxtamembrane region. 24798328 0 31 theme Low 0:2 arg1 lipoprotein 12:22 Low density lipoprotein 0:22 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 0 32 theme lipoprotein 12:22 arg1 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 1 33 theme density 95:101 arg1 lipoprotein 103:113 low density lipoprotein 91:113 The low density lipoprotein receptor (LDLR) 87:129 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 4 34 theme LDLR-related 577:588 arg1 receptors 590:598 several LDLR-related receptors 569:598 several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 569:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 6 35 gly O-glycosylation 901:915 arg2 position 925:932 position -1 925:935 position -1 of the first cysteine residue of most repeats 925:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 35 gly O-glycosylation 901:915 arg2 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 0 36 theme class 33:37 arg1 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 9 37 theme regions 1372:1378 arg1 glycosylation 1344:1356 glycosylation 1344:1356 glycosylation of the linker regions of the LDLR class A repeats 1344:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 6 38 theme O-glycosylation 901:915 arg1 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 10 39 from expression 1494:1503 arg1 cells 1523:1527 HEK293 cells 1516:1527 HEK293 cells 1516:1527 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 4 40 theme LDLR 545:548 arg1 repeats 558:564 the characteristic LDLR class A repeats 526:564 the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 526:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 10 41 theme HEK293 1516:1521 arg1 cells 1523:1527 HEK293 cells 1516:1527 HEK293 cells 1516:1527 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 4 42 theme A 556:556 arg1 repeats 558:564 the characteristic LDLR class A repeats 526:564 the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 526:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 10 43 theme LDLR 1508:1511 arg1 expression 1494:1503 expression 1494:1503 expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions 1494:1640 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 6 44 gly glycosylated 1016:1027 arg2 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 44 gly glycosylated 1016:1027 arg1 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 44 gly glycosylated 1016:1027 arg1 cells 1007:1011 wild-type CHO cells 993:1011 wild-type CHO cells 993:1011 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 2 45 theme cell 329:332 arg1 surface 334:340 the cell surface 325:340 the cell surface 325:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 9 46 theme class 1392:1396 arg1 repeats 1400:1406 the LDLR class A repeats 1383:1406 the LDLR class A repeats 1383:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 4 47 theme " 621:621 arg1 strategy 647:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 4 48 gly O-glycosylation 474:488 arg2 sites 490:494 O-glycosylation sites 474:494 O-glycosylation sites 474:494 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 5 49 from sites 718:722 arg1 LDLR 739:742 recombinant LDLR 727:742 recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells 727:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 7 50 from glycosites 1079:1088 arg1 regions 1100:1106 linker regions 1093:1106 linker regions of LDLR class A repeats 1093:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 6 51 gly sialylated 1046:1055 arg1 structure 1064:1072 the typical sialylated core 1 structure 1034:1072 the typical sialylated core 1 structure 1034:1072 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 9 52 theme in 1411:1412 arg1 assays 1427:1432 in vitro enzyme assays 1411:1432 in vitro enzyme assays 1411:1432 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 6 53 theme cysteine 950:957 arg1 residue 959:965 the first cysteine residue 940:965 the first cysteine residue of most repeats 940:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 53 theme cysteine 950:957 arg1 repeats 975:981 most repeats 970:981 most repeats 970:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 10 54 theme isoform 1564:1570 arg1 knock-out 1536:1544 knock-out 1536:1544 knock-out of the GalNAc-T11 isoform 1536:1570 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 6 55 theme class 849:853 arg1 A 855:855 LDLR class A 844:855 LDLR class A repeats 844:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 9 56 theme enzyme 1420:1425 arg1 assays 1427:1432 in vitro enzyme assays 1411:1432 in vitro enzyme assays 1411:1432 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 7 57 theme A 1122:1122 arg1 repeats 1124:1130 LDLR class A repeats 1111:1130 LDLR class A repeats 1111:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 5 58 gly O-glycosylation 702:716 arg2 sites 718:722 O-glycosylation sites 702:722 O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells 702:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 1 59 theme cholesterol 146:156 arg1 homeostasis 158:168 cholesterol homeostasis 146:168 cholesterol homeostasis 146:168 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 8 60 theme large 1250:1254 arg1 family 1256:1261 a large family 1248:1261 a large family of polypeptide GalNAc transferases 1248:1296 O-Glycosylation is controlled by a large family of polypeptide GalNAc transferases. 24798328 7 61 theme LDLR 1111:1114 arg1 A 1122:1122 LDLR class A 1111:1122 LDLR class A repeats 1111:1130 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 6 62 theme linker 821:826 arg1 regions 828:834 the short linker regions 811:834 the short linker regions between LDLR class A repeats 811:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 9 63 theme major 1446:1450 arg1 role 1452:1455 a major role 1444:1455 a major role of GalNAc-T11 1444:1469 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 2 64 theme stable 304:309 arg1 expression 311:320 stable expression 304:320 stable expression at the cell surface 304:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 6 65 theme wild-type 993:1001 arg1 cells 1007:1011 wild-type CHO cells 993:1011 wild-type CHO cells 993:1011 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 0 66 from regions 78:84 arg1 O-glycosylated 53:66 O-glycosylated 53:66 O-glycosylated 53:66 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 8 67 theme GalNAc 1278:1283 arg1 transferases 1285:1296 polypeptide GalNAc transferases 1266:1296 polypeptide GalNAc transferases 1266:1296 O-Glycosylation is controlled by a large family of polypeptide GalNAc transferases. 24798328 5 68 theme HEK293 754:759 arg1 SimpleCells 761:771 HEK293 SimpleCells 754:771 HEK293 SimpleCells 754:771 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 4 69 theme O-glycosylation 474:488 arg1 sites 490:494 O-glycosylation sites 474:494 O-glycosylation sites 474:494 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 10 70 theme glycosylation 1596:1608 arg1 loss 1588:1591 the loss 1584:1591 the loss of glycosylation of three of four linker regions 1584:1640 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 1 71 theme lipoprotein 103:113 arg1 LDLR 125:128 LDLR 125:128 LDLR 125:128 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 1 71 theme lipoprotein 103:113 arg1 crucial 134:140 crucial 134:140 crucial 134:140 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 1 71 theme lipoprotein 103:113 arg1 receptor 115:122 The low density lipoprotein receptor 87:122 The low density lipoprotein receptor (LDLR) 87:129 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 6 72 theme repeats 975:981 arg1 residue 959:965 the first cysteine residue 940:965 the first cysteine residue of most repeats 940:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 72 theme repeats 975:981 arg1 repeats 975:981 most repeats 970:981 most repeats 970:981 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 5 73 theme recombinant 727:737 arg1 LDLR 739:742 recombinant LDLR 727:742 recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells 727:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 2 74 theme transmembrane 248:260 arg1 LDLR 231:234 LDLR 231:234 LDLR 231:234 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 2 74 theme transmembrane 248:260 arg1 protein 262:268 a type I transmembrane protein 239:268 a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface 239:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 9 75 dep in 1411:1412 arg1 vitro 1414:1418 vitro 1414:1418 vitro 1414:1418 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 2 76 theme type 241:244 arg1 LDLR 231:234 LDLR 231:234 LDLR 231:234 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 2 76 theme type 241:244 arg1 protein 262:268 a type I transmembrane protein 239:268 a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface 239:340 LDLR is a type I transmembrane protein that requires O-glycosylation for stable expression at the cell surface. 24798328 1 77 from deficiency 174:183 arg1 functions 193:201 LDLR functions 188:201 LDLR functions 188:201 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 6 78 theme sialylated 1046:1055 arg1 structure 1064:1072 the typical sialylated core 1 structure 1034:1072 the typical sialylated core 1 structure 1034:1072 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 79 contain contain 865:871 arg1 regions 828:834 the short linker regions 811:834 the short linker regions between LDLR class A repeats 811:863 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 6 79 contain contain 865:871 arg2 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 4 80 theme characteristic 530:543 arg1 repeats 558:564 the characteristic LDLR class A repeats 526:564 the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 526:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 0 81 gly O-glycosylated 53:66 arg2 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 0 81 gly O-glycosylated 53:66 arg1 regions 78:84 linker regions 71:84 linker regions 71:84 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 0 81 gly O-glycosylated 53:66 arg1 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 9 82 gly glycosylation 1344:1356 arg1 regions 1372:1378 the linker regions 1361:1378 the linker regions of the LDLR class A repeats 1361:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 5 83 theme wild-type 781:789 arg1 cells 791:795 CHO wild-type cells 777:795 CHO wild-type cells 777:795 Herein, we have systematically characterized O-glycosylation sites on recombinant LDLR shed from HEK293 SimpleCells and CHO wild-type cells. 24798328 9 84 theme GalNAc-T11 1460:1469 arg1 role 1452:1455 a major role 1444:1455 a major role of GalNAc-T11 1444:1469 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 4 85 theme linker 503:508 arg1 regions 510:516 the linker regions 499:516 the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 499:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 7 86 theme homologous 1193:1202 arg1 receptors 1204:1212 other homologous receptors 1187:1212 other homologous receptors 1187:1212 The glycosites in linker regions of LDLR class A repeats are conserved in LDLR from man to Xenopus and found in other homologous receptors. 24798328 0 87 theme density 4:10 arg1 lipoprotein 12:22 Low density lipoprotein 0:22 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 10 88 theme GalNAc-T11 1553:1562 arg1 isoform 1564:1570 the GalNAc-T11 isoform 1549:1570 the GalNAc-T11 isoform 1549:1570 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 4 89 theme several 569:575 arg1 receptors 590:598 several LDLR-related receptors 569:598 several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 569:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 3 90 theme juxtamembrane 429:441 arg1 region 443:448 the juxtamembrane region 425:448 the juxtamembrane region 425:448 It has previously been suggested that LDLR O-glycosylation is found N-terminal to the juxtamembrane region. 24798328 0 91 theme receptor 24:31 arg1 repeats 41:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats 0:47 Low density lipoprotein receptor class A repeats are O-glycosylated in linker regions. 24798328 1 92 theme low 91:93 arg1 lipoprotein 103:113 low density lipoprotein 91:113 The low density lipoprotein receptor (LDLR) 87:129 The low density lipoprotein receptor (LDLR) is crucial for cholesterol homeostasis and deficiency in LDLR functions cause hypercholesterolemia. 24798328 4 93 from repeats 558:564 arg1 receptors 590:598 several LDLR-related receptors 569:598 several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 569:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 9 94 theme linker 1365:1370 arg1 regions 1372:1378 the linker regions 1361:1378 the linker regions of the LDLR class A repeats 1361:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 4 95 theme class 550:554 arg1 repeats 558:564 the characteristic LDLR class A repeats 526:564 the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy 526:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 9 96 theme repeats 1400:1406 arg1 regions 1372:1378 the linker regions 1361:1378 the linker regions of the LDLR class A repeats 1361:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 6 97 theme conserved 891:899 arg1 site 917:920 an evolutionarily conserved O-glycosylation site 873:920 an evolutionarily conserved O-glycosylation site 873:920 We find that the short linker regions between LDLR class A repeats contain an evolutionarily conserved O-glycosylation site at position -1 of the first cysteine residue of most repeats, which in wild-type CHO cells is glycosylated with the typical sialylated core 1 structure. 24798328 9 98 theme LDLR 1387:1390 arg1 repeats 1400:1406 the LDLR class A repeats 1383:1406 the LDLR class A repeats 1383:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 4 99 theme O-glycoproteome 623:637 arg1 strategy 647:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 24798328 9 100 theme A 1398:1398 arg1 repeats 1400:1406 the LDLR class A repeats 1383:1406 the LDLR class A repeats 1383:1406 Probing into which isoform(s) contributed to glycosylation of the linker regions of the LDLR class A repeats by in vitro enzyme assays suggested a major role of GalNAc-T11. 24798328 10 101 theme linker 1627:1632 arg1 regions 1634:1640 three of four linker regions 1613:1640 three of four linker regions 1613:1640 This was supported by expression of LDLR in HEK293 cells, where knock-out of the GalNAc-T11 isoform resulted in the loss of glycosylation of three of four linker regions. 24798328 4 102 theme shotgun 639:645 arg1 strategy 647:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 the "SimpleCell" O-glycoproteome shotgun strategy 606:654 Recently we identified O-glycosylation sites in the linker regions between the characteristic LDLR class A repeats in several LDLR-related receptors using the "SimpleCell" O-glycoproteome shotgun strategy. 28921966 8 0 theme peptides 1499:1506 arg1 library 1472:1478 a library 1470:1478 a library of short synthetic peptides 1470:1506 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 4 1 theme intact 705:710 arg1 glycopeptides 712:724 intact glycopeptides 705:724 intact glycopeptides 705:724 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 2 2 theme glycoprotein 354:365 arg1 analysis 367:374 glycoprotein analysis 354:374 glycoprotein analysis 354:374 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 0 3 theme Site 70:73 arg1 Mapping 75:81 One-Pot Site Mapping 62:81 One-Pot Site Mapping 62:81 Tool for Rapid Analysis of Glycopeptide by Permethylation via One-Pot Site Mapping and Glycan Analysis. 28921966 10 4 theme glycan 1835:1840 arg1 structures 1842:1851 O-linked glycan structures 1826:1851 O-linked glycan structures 1826:1851 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 7 5 theme high 1120:1123 arg1 system 1147:1152 a high resolution tandem-MSn system 1118:1152 a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1118:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 6 6 theme single 1072:1077 arg1 procedure 1092:1100 a single experimental procedure 1070:1100 a single experimental procedure 1070:1100 Remarkably, the glycan patterns, glycosylation site, and their occupancy by N-glycans are all detected and identified in a single experimental procedure. 28921966 0 7 theme Glycan 87:92 arg1 Analysis 94:101 Glycan Analysis 87:101 Glycan Analysis 87:101 Tool for Rapid Analysis of Glycopeptide by Permethylation via One-Pot Site Mapping and Glycan Analysis. 28921966 8 8 theme basic 1409:1413 arg1 conditions 1430:1439 the basic permethylation conditions 1405:1439 the basic permethylation conditions 1405:1439 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 2 9 gly glycopeptide 413:424 arg2 glycopeptide 413:424 glycopeptide analysis 413:433 glycopeptide analysis 413:433 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 3 10 from prokaryotes 528:538 arg1 glycans 469:475 glycans 469:475 glycans 469:475 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 9 11 theme data 1686:1689 arg1 interpretation 1662:1675 the interpretation 1658:1675 the interpretation of MS/MS data 1658:1689 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 9 12 theme chemical 1595:1602 arg1 transformations 1604:1618 simple, limited, and predictable chemical transformations 1562:1618 simple, limited, and predictable chemical transformations on peptides 1562:1630 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 1 13 theme glycopeptides 252:264 arg1 analysis 302:309 their tandem mass spectrometric analysis 270:309 their tandem mass spectrometric analysis 270:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 1 13 theme glycopeptides 252:264 arg1 permethylation 234:247 permethylation 234:247 permethylation of glycopeptides 234:264 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 9 14 theme MS/MS 1680:1684 arg1 data 1686:1689 MS/MS data 1680:1689 MS/MS data 1680:1689 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 3 15 theme enzymatic 564:572 arg1 release 586:592 enzymatic or chemical release 564:592 enzymatic or chemical release 564:592 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 10 16 link O-linked 1826:1833 arg1 structures 1842:1851 O-linked glycan structures 1826:1851 O-linked glycan structures 1826:1851 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 7 17 theme high-energy 1195:1205 arg1 HCD 1231:1233 high-energy collision dissociation (HCD) 1195:1234 high-energy collision dissociation (HCD) 1195:1234 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 5 18 theme method 941:946 arg1 feasibility 922:932 the feasibility 918:932 the feasibility of the method 918:946 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 6 19 theme experimental 1079:1090 arg1 procedure 1092:1100 a single experimental procedure 1070:1100 a single experimental procedure 1070:1100 Remarkably, the glycan patterns, glycosylation site, and their occupancy by N-glycans are all detected and identified in a single experimental procedure. 28921966 7 20 with system 1147:1152 arg1 methodologies 1173:1185 fragmentation methodologies 1159:1185 fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1159:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 20 with system 1147:1152 arg1 HCD 1231:1233 high-energy collision dissociation (HCD) 1195:1234 high-energy collision dissociation (HCD) 1195:1234 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 20 with system 1147:1152 arg1 collision 1240:1248 collision 1240:1248 collision 1240:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 21 theme glycan 1316:1321 arg1 structures 1323:1332 the glycan structures 1312:1332 the glycan structures attached to the peptides 1312:1357 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 1 22 theme protein 150:156 arg1 glycosylation 158:170 protein glycosylation 150:170 protein glycosylation 150:170 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 7 23 theme collision 1207:1215 arg1 HCD 1231:1233 high-energy collision dissociation (HCD) 1195:1234 high-energy collision dissociation (HCD) 1195:1234 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 9 24 theme limited 1570:1576 arg1 transformations 1604:1618 simple, limited, and predictable chemical transformations 1562:1618 simple, limited, and predictable chemical transformations on peptides 1562:1630 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 1 25 theme glycosylation 158:170 arg1 analysis 138:145 the analysis 134:145 the analysis of protein glycosylation 134:170 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 10 26 theme β 1768:1768 arg1 elimination 1770:1780 β elimination 1768:1780 β elimination 1768:1780 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 1 27 from challenges 120:129 arg1 analysis 138:145 the analysis 134:145 the analysis of protein glycosylation 134:170 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 4 28 theme protease 751:758 arg1 digest 760:765 controlled protease digest 740:765 controlled protease digest 740:765 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 2 29 theme glycan 391:396 arg1 sequencing 398:407 glycan sequencing 391:407 glycan sequencing 391:407 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 8 30 theme short 1483:1487 arg1 peptides 1499:1506 short synthetic peptides 1483:1506 short synthetic peptides 1483:1506 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 4 31 theme controlled 740:749 arg1 digest 760:765 controlled protease digest 740:765 controlled protease digest 740:765 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 5 32 theme bovine 879:884 arg1 models 896:901 models 896:901 models 896:901 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 32 theme bovine 879:884 arg1 fetuin 886:891 bovine fetuin 879:891 bovine fetuin 879:891 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 32 theme bovine 879:884 arg1 B 853:853 bovine RNase B 840:853 bovine RNase B 840:853 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 32 theme bovine 879:884 arg1 transferrin 862:872 human transferrin 856:872 human transferrin 856:872 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 7 33 theme fragmentation 1159:1171 arg1 methodologies 1173:1185 fragmentation methodologies 1159:1185 fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1159:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 33 theme fragmentation 1159:1171 arg1 HCD 1231:1233 high-energy collision dissociation (HCD) 1195:1234 high-energy collision dissociation (HCD) 1195:1234 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 33 theme fragmentation 1159:1171 arg1 collision 1240:1248 collision 1240:1248 collision 1240:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 1 34 theme tandem 276:281 arg1 analysis 302:309 their tandem mass spectrometric analysis 270:309 their tandem mass spectrometric analysis 270:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 4 35 gly glycopeptides 712:724 arg2 glycopeptides 712:724 intact glycopeptides 705:724 intact glycopeptides 705:724 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 1 36 gly glycopeptides 252:264 arg2 glycopeptides 252:264 glycopeptides 252:264 glycopeptides 252:264 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 8 37 theme natural 1379:1385 arg1 acids 1393:1397 20 natural amino acids 1376:1397 20 natural amino acids 1376:1397 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 5 38 theme RNase 847:851 arg1 models 896:901 models 896:901 models 896:901 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 38 theme RNase 847:851 arg1 fetuin 886:891 bovine fetuin 879:891 bovine fetuin 879:891 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 38 theme RNase 847:851 arg1 B 853:853 bovine RNase B 840:853 bovine RNase B 840:853 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 38 theme RNase 847:851 arg1 transferrin 862:872 human transferrin 856:872 human transferrin 856:872 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 2 39 theme single 440:445 arg1 experiment 447:456 a single experiment 438:456 a single experiment 438:456 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 1 40 theme mass 283:286 arg1 analysis 302:309 their tandem mass spectrometric analysis 270:309 their tandem mass spectrometric analysis 270:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 3 41 with glycans 469:475 arg1 linkages 500:507 unique glycosidic linkages 482:507 unique glycosidic linkages 482:507 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 3 42 theme chemical 577:584 arg1 release 586:592 enzymatic or chemical release 564:592 enzymatic or chemical release 564:592 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 9 43 from transformations 1604:1618 arg1 peptides 1623:1630 peptides 1623:1630 peptides 1623:1630 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 3 44 theme unique 482:487 arg1 linkages 500:507 unique glycosidic linkages 482:507 unique glycosidic linkages 482:507 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 0 45 theme Glycopeptide 27:38 arg1 Analysis 15:22 Rapid Analysis 9:22 Rapid Analysis of Glycopeptide by Permethylation 9:56 Tool for Rapid Analysis of Glycopeptide by Permethylation via One-Pot Site Mapping and Glycan Analysis. 28921966 1 46 theme spectrometric 288:300 arg1 analysis 302:309 their tandem mass spectrometric analysis 270:309 their tandem mass spectrometric analysis 270:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 8 47 theme acids 1393:1397 arg1 behavior 1364:1371 The behavior 1360:1371 The behavior of 20 natural amino acids under the basic permethylation conditions 1360:1439 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 7 48 theme structures 1323:1332 arg1 sequence 1300:1307 the complete sequence 1287:1307 the complete sequence of the glycan structures attached to the peptides 1287:1357 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 49 theme complete 1291:1298 arg1 sequence 1300:1307 the complete sequence 1287:1307 the complete sequence of the glycan structures attached to the peptides 1287:1357 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 50 from Acquisition 1103:1113 arg1 system 1147:1152 a high resolution tandem-MSn system 1118:1152 a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1118:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 2 51 gly glycoprotein 354:365 arg1 glycoprotein 354:365 glycoprotein analysis 354:374 glycoprotein analysis 354:374 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 8 52 theme amino 1387:1391 arg1 acids 1393:1397 20 natural amino acids 1376:1397 20 natural amino acids 1376:1397 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 5 53 theme human 856:860 arg1 models 896:901 models 896:901 models 896:901 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 53 theme human 856:860 arg1 fetuin 886:891 bovine fetuin 879:891 bovine fetuin 879:891 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 53 theme human 856:860 arg1 B 853:853 bovine RNase B 840:853 bovine RNase B 840:853 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 53 theme human 856:860 arg1 transferrin 862:872 human transferrin 856:872 human transferrin 856:872 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 1 54 theme comprehensive 193:205 arg1 tool 221:224 a comprehensive and universal tool 191:224 a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis 191:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 7 55 theme resolution 1125:1134 arg1 system 1147:1152 a high resolution tandem-MSn system 1118:1152 a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1118:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 56 theme dissociation 1217:1228 arg1 HCD 1231:1233 high-energy collision dissociation (HCD) 1195:1234 high-energy collision dissociation (HCD) 1195:1234 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 6 57 theme glycan 965:970 arg1 patterns 972:979 the glycan patterns 961:979 the glycan patterns 961:979 Remarkably, the glycan patterns, glycosylation site, and their occupancy by N-glycans are all detected and identified in a single experimental procedure. 28921966 10 58 theme permethylated 1713:1725 arg1 O-glycans 1727:1735 permethylated O-glycans 1713:1735 permethylated O-glycans in unreduced form (released by β elimination) 1713:1781 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 7 59 theme tandem-MSn 1136:1145 arg1 system 1147:1152 a high resolution tandem-MSn system 1118:1152 a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision 1118:1248 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 5 60 used used 835:838 arg2 We 832:833 We 832:833 We 832:833 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 4 61 theme glycopeptides 712:724 arg1 permethylation 687:700 the permethylation 683:700 the permethylation of intact glycopeptides, obtained via controlled protease digest, 683:766 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 8 62 theme permethylation 1415:1428 arg1 conditions 1430:1439 the basic permethylation conditions 1405:1439 the basic permethylation conditions 1405:1439 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 4 63 theme mass 813:816 arg1 spectrometry 818:829 advanced mass spectrometry 804:829 advanced mass spectrometry 804:829 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 7 64 attach attached 1334:1341 arg2 structures 1323:1332 the glycan structures 1312:1332 the glycan structures attached to the peptides 1312:1357 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 7 64 attach attached 1334:1341 arg1 peptides 1350:1357 the peptides 1346:1357 the peptides 1346:1357 Acquisition on a high resolution tandem-MSn system with fragmentation methodologies such as high-energy collision dissociation (HCD) and collision induced dissociation (CID), provided the complete sequence of the glycan structures attached to the peptides. 28921966 4 65 theme advanced 804:811 arg1 spectrometry 818:829 advanced mass spectrometry 804:829 advanced mass spectrometry 804:829 Here we present a strategy for the permethylation of intact glycopeptides, obtained via controlled protease digest, and their characterization by using advanced mass spectrometry. 28921966 6 66 gly glycosylation 982:994 arg2 site 996:999 glycosylation site 982:999 glycosylation site 982:999 Remarkably, the glycan patterns, glycosylation site, and their occupancy by N-glycans are all detected and identified in a single experimental procedure. 28921966 3 67 theme glycosidic 489:498 arg1 linkages 500:507 unique glycosidic linkages 482:507 unique glycosidic linkages 482:507 Moreover, glycans with unique glycosidic linkages, particularly from prokaryotes, which are resistant to enzymatic or chemical release, could also be detected and analyzed by this methodology. 28921966 9 68 theme predictable 1583:1593 arg1 transformations 1604:1618 simple, limited, and predictable chemical transformations 1562:1618 simple, limited, and predictable chemical transformations on peptides 1562:1630 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 10 69 from O-glycans 1727:1735 arg1 form 1750:1753 unreduced form 1740:1753 unreduced form 1740:1753 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 8 70 theme synthetic 1489:1497 arg1 peptides 1499:1506 short synthetic peptides 1483:1506 short synthetic peptides 1483:1506 The behavior of 20 natural amino acids under the basic permethylation conditions was probed by permethylating a library of short synthetic peptides. 28921966 5 71 theme bovine 840:845 arg1 models 896:901 models 896:901 models 896:901 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 71 theme bovine 840:845 arg1 fetuin 886:891 bovine fetuin 879:891 bovine fetuin 879:891 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 71 theme bovine 840:845 arg1 B 853:853 bovine RNase B 840:853 bovine RNase B 840:853 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 5 71 theme bovine 840:845 arg1 transferrin 862:872 human transferrin 856:872 human transferrin 856:872 We used bovine RNase B, human transferrin, and bovine fetuin as models to demonstrate the feasibility of the method. 28921966 1 72 theme universal 211:219 arg1 tool 221:224 a comprehensive and universal tool 191:224 a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis 191:309 To overcome the challenges in the analysis of protein glycosylation, we have developed a comprehensive and universal tool through permethylation of glycopeptides and their tandem mass spectrometric analysis. 28921966 0 73 theme One-Pot 62:68 arg1 Mapping 75:81 One-Pot Site Mapping 62:81 One-Pot Site Mapping 62:81 Tool for Rapid Analysis of Glycopeptide by Permethylation via One-Pot Site Mapping and Glycan Analysis. 28921966 2 74 theme glycopeptide 413:424 arg1 analysis 426:433 glycopeptide analysis 413:433 glycopeptide analysis 413:433 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 9 75 theme simple 1562:1567 arg1 transformations 1604:1618 simple, limited, and predictable chemical transformations 1562:1618 simple, limited, and predictable chemical transformations on peptides 1562:1630 Our studies indicate that the permethylation imparts simple, limited, and predictable chemical transformations on peptides and do not interfere with the interpretation of MS/MS data. 28921966 6 76 theme glycosylation 982:994 arg1 site 996:999 glycosylation site 982:999 glycosylation site 982:999 Remarkably, the glycan patterns, glycosylation site, and their occupancy by N-glycans are all detected and identified in a single experimental procedure. 28921966 2 77 contain has 324:326 arg2 potential 332:340 the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment 328:456 the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment 328:456 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 2 77 contain has 324:326 arg1 method 317:322 This method 312:322 This method 312:322 This method has the potential to simplify glycoprotein analysis by integrating glycan sequencing and glycopeptide analysis in a single experiment. 28921966 0 78 theme Rapid 9:13 arg1 Analysis 15:22 Rapid Analysis 9:22 Rapid Analysis of Glycopeptide by Permethylation 9:56 Tool for Rapid Analysis of Glycopeptide by Permethylation via One-Pot Site Mapping and Glycan Analysis. 28921966 10 79 theme unreduced 1740:1748 arg1 form 1750:1753 unreduced form 1740:1753 unreduced form 1740:1753 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 28921966 10 80 theme O-linked 1826:1833 arg1 structures 1842:1851 O-linked glycan structures 1826:1851 O-linked glycan structures 1826:1851 In addition to this, permethylated O-glycans in unreduced form (released by β elimination) were also detected, allowing us to profile O-linked glycan structures simultaneously. 26687240 1 0 theme activities 137:146 arg1 regulators 114:123 important regulators 104:123 important regulators of antibody activities and half-lives 104:161 Glycans serve as important regulators of antibody activities and half-lives. 26687240 1 0 theme activities 137:146 arg1 Glycans 87:93 Glycans 87:93 Glycans 87:93 Glycans serve as important regulators of antibody activities and half-lives. 26687240 4 1 with patient 660:666 arg1 mutation 678:685 PGM3 mutation 673:685 PGM3 mutation 673:685 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 4 1 with patient 660:666 arg1 dermatitis 713:722 atopic dermatitis 706:722 atopic dermatitis 706:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 8 2 theme previous 1167:1174 arg1 knowledge 1176:1184 previous knowledge 1167:1184 previous knowledge of IgE glycosylation 1167:1205 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 7 3 theme similar 1073:1079 arg1 profiles 1095:1102 similar glycosylation profiles 1073:1102 similar glycosylation profiles between the two IgE samples 1073:1130 The results showed similar glycosylation profiles between the two IgE samples. 26687240 6 4 theme tri-antennary 925:937 arg1 glycans 939:945 tri-antennary glycans 925:945 tri-antennary glycans 925:945 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 5 located observed 964:971 arg1 Asn252 986:991 Asn252 986:991 Asn252 986:991 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 5 located observed 964:971 arg1 Asn99 976:980 Asn99 976:980 Asn99 976:980 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 5 located observed 964:971 arg2 glycans 939:945 tri-antennary glycans 925:945 tri-antennary glycans 925:945 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 3 6 with patient 381:387 arg1 syndrome 412:419 a novel hyper IgE syndrome 394:419 a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 394:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 4 7 theme two-step 557:564 arg1 method 566:571 A two-step method 555:571 A two-step method 555:571 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 0 8 theme PGM3 72:75 arg1 mutation 77:84 PGM3 mutation 72:84 PGM3 mutation 72:84 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 5 9 used used 782:785 arg2 strategy 769:776 a glycoproteomic strategy 752:776 a glycoproteomic strategy 752:776 Then, a glycoproteomic strategy was used to study the site-specific glycosylation. 26687240 3 10 theme sugar 508:512 arg1 UDP-GlcNAc 494:503 UDP-GlcNAc 494:503 UDP-GlcNAc 494:503 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 3 10 theme sugar 508:512 arg1 donor 514:518 a sugar donor 506:518 a sugar donor widely required for glycosylation 506:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 8 11 theme hyper 1308:1312 arg1 syndrome 1318:1325 PGM3 related hyper IgE syndrome 1295:1325 PGM3 related hyper IgE syndrome 1295:1325 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 5 12 theme site-specific 800:812 arg1 glycosylation 814:826 the site-specific glycosylation 796:826 the site-specific glycosylation 796:826 Then, a glycoproteomic strategy was used to study the site-specific glycosylation. 26687240 1 13 theme half-lives 152:161 arg1 regulators 114:123 important regulators 104:123 important regulators of antibody activities and half-lives 104:161 Glycans serve as important regulators of antibody activities and half-lives. 26687240 1 13 theme half-lives 152:161 arg1 Glycans 87:93 Glycans 87:93 Glycans 87:93 Glycans serve as important regulators of antibody activities and half-lives. 26687240 7 14 theme glycosylation 1081:1093 arg1 profiles 1095:1102 similar glycosylation profiles 1073:1102 similar glycosylation profiles between the two IgE samples 1073:1130 The results showed similar glycosylation profiles between the two IgE samples. 26687240 3 15 attach linked 421:426 arg1 mutations 431:439 mutations 431:439 mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 431:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 3 15 attach linked 421:426 arg2 syndrome 412:419 a novel hyper IgE syndrome 394:419 a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 394:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 8 16 theme related 1300:1306 arg1 syndrome 1318:1325 PGM3 related hyper IgE syndrome 1295:1325 PGM3 related hyper IgE syndrome 1295:1325 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 4 17 from less 619:622 arg1 subject 737:743 a control subject 727:743 a control subject 727:743 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 4 17 from less 619:622 arg1 samples 606:612 two IgE samples 598:612 two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis 598:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 6 18 theme complex 1014:1020 arg1 glycans 1022:1028 non-fucosylated complex glycans 998:1028 non-fucosylated complex glycans 998:1028 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 7 19 theme IgE 1120:1122 arg1 samples 1124:1130 the two IgE samples 1112:1130 the two IgE samples 1112:1130 The results showed similar glycosylation profiles between the two IgE samples. 26687240 2 20 theme function 292:299 arg1 relationships 301:313 its glycan structure function relationships 271:313 its glycan structure function relationships 271:313 IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. 26687240 6 21 theme non-fucosylated 998:1012 arg1 glycans 1022:1028 non-fucosylated complex glycans 998:1028 non-fucosylated complex glycans 998:1028 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 3 22 theme hyper 402:406 arg1 syndrome 412:419 a novel hyper IgE syndrome 394:419 a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 394:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 0 23 theme Glycoproteomic 0:13 arg1 studies 15:21 Glycoproteomic studies 0:21 Glycoproteomic studies of IgE from a novel hyper IgE syndrome 0:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 8 24 theme IgE 1189:1191 arg1 glycosylation 1193:1205 IgE glycosylation 1189:1205 IgE glycosylation 1189:1205 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 2 25 theme structure 282:290 arg1 relationships 301:313 its glycan structure function relationships 271:313 its glycan structure function relationships 271:313 IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. 26687240 4 26 theme atopic 706:711 arg1 dermatitis 713:722 atopic dermatitis 706:722 atopic dermatitis 706:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 3 27 theme IgE 408:410 arg1 syndrome 412:419 a novel hyper IgE syndrome 394:419 a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 394:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 3 28 theme IgE 356:358 arg1 glycosylation 360:372 the site specific IgE glycosylation 338:372 the site specific IgE glycosylation 338:372 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 2 29 theme glycan 275:280 arg1 relationships 301:313 its glycan structure function relationships 271:313 its glycan structure function relationships 271:313 IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. 26687240 5 30 theme glycoproteomic 754:767 arg1 strategy 769:776 a glycoproteomic strategy 752:776 a glycoproteomic strategy 752:776 Then, a glycoproteomic strategy was used to study the site-specific glycosylation. 26687240 8 31 theme glycosylation 1193:1205 arg1 knowledge 1176:1184 previous knowledge 1167:1184 previous knowledge of IgE glycosylation 1167:1205 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 6 32 located found 850:854 arg1 Asn264 859:864 Asn264 859:864 Asn264 859:864 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 32 located found 850:854 arg2 glycosylation 832:844 No glycosylation 829:844 No glycosylation 829:844 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 3 33 theme novel 396:400 arg1 syndrome 412:419 a novel hyper IgE syndrome 394:419 a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 394:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 4 34 with patient 693:699 arg1 mutation 678:685 PGM3 mutation 673:685 PGM3 mutation 673:685 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 4 34 with patient 693:699 arg1 dermatitis 713:722 atopic dermatitis 706:722 atopic dermatitis 706:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 0 35 from syndrome 53:60 arg1 studies 15:21 Glycoproteomic studies 0:21 Glycoproteomic studies of IgE from a novel hyper IgE syndrome 0:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 0 36 theme IgE 26:28 arg1 studies 15:21 Glycoproteomic studies 0:21 Glycoproteomic studies of IgE from a novel hyper IgE syndrome 0:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 4 37 theme IgE 602:604 arg1 subject 737:743 a control subject 727:743 a control subject 727:743 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 4 37 theme IgE 602:604 arg1 samples 606:612 two IgE samples 598:612 two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis 598:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 0 38 theme hyper 43:47 arg1 syndrome 53:60 a novel hyper IgE syndrome 35:60 a novel hyper IgE syndrome 35:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 6 39 theme mannose 879:885 arg1 glycans 887:893 high mannose glycans 874:893 high mannose glycans 874:893 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 8 40 theme PGM3 1295:1298 arg1 syndrome 1318:1325 PGM3 related hyper IgE syndrome 1295:1325 PGM3 related hyper IgE syndrome 1295:1325 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 6 41 located detected 1035:1042 arg2 glycans 1022:1028 non-fucosylated complex glycans 998:1028 non-fucosylated complex glycans 998:1028 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 41 located detected 1035:1042 arg1 Asn99 1047:1051 Asn99 1047:1051 Asn99 1047:1051 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 3 42 from mutations 431:439 arg1 enzyme 462:467 an enzyme 459:467 an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation 459:552 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 3 42 from mutations 431:439 arg1 PGM3 444:447 PGM3 444:447 PGM3 444:447 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26687240 0 43 theme novel 37:41 arg1 syndrome 53:60 a novel hyper IgE syndrome 35:60 a novel hyper IgE syndrome 35:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 6 44 theme high 874:877 arg1 glycans 887:893 high mannose glycans 874:893 high mannose glycans 874:893 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 0 45 attach linked 62:67 arg1 mutation 77:84 PGM3 mutation 72:84 PGM3 mutation 72:84 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 0 45 attach linked 62:67 arg2 studies 15:21 Glycoproteomic studies 0:21 Glycoproteomic studies of IgE from a novel hyper IgE syndrome 0:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 6 46 gly glycosylation 832:844 arg2 Asn264 859:864 Asn264 859:864 Asn264 859:864 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 46 gly glycosylation 832:844 arg1 Asn264 859:864 Asn264 859:864 Asn264 859:864 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 47 located detected 905:912 arg1 Asn275 917:922 Asn275 917:922 Asn275 917:922 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 6 47 located detected 905:912 arg2 glycans 887:893 high mannose glycans 874:893 high mannose glycans 874:893 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 4 48 theme control 729:735 arg1 subject 737:743 a control subject 727:743 a control subject 727:743 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 4 48 theme control 729:735 arg1 samples 606:612 two IgE samples 598:612 two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis 598:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 1 49 theme important 104:112 arg1 regulators 114:123 important regulators 104:123 important regulators of antibody activities and half-lives 104:161 Glycans serve as important regulators of antibody activities and half-lives. 26687240 1 49 theme important 104:112 arg1 Glycans 87:93 Glycans 87:93 Glycans 87:93 Glycans serve as important regulators of antibody activities and half-lives. 26687240 2 50 gly glycosylated 188:199 arg1 IgE 164:166 IgE 164:166 IgE 164:166 IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. 26687240 6 51 gly non-fucosylated 998:1012 arg1 glycans 1022:1028 non-fucosylated complex glycans 998:1028 non-fucosylated complex glycans 998:1028 No glycosylation was found at Asn264, whilst high mannose glycans were only detected at Asn275, tri-antennary glycans were exclusively observed at Asn99 and Asn252, and non-fucosylated complex glycans were detected at Asn99. 26687240 2 52 theme other 232:236 arg1 antibodies 238:247 other antibodies 232:247 other antibodies 232:247 IgE is the most heavily glycosylated antibody, but in comparison to other antibodies little is known about its glycan structure function relationships. 26687240 8 53 theme healthy 1266:1272 arg1 control 1274:1280 healthy control 1266:1280 healthy control 1266:1280 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 8 54 theme IgE 1314:1316 arg1 syndrome 1318:1325 PGM3 related hyper IgE syndrome 1295:1325 PGM3 related hyper IgE syndrome 1295:1325 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 4 55 theme PGM3 673:676 arg1 mutation 678:685 PGM3 mutation 673:685 PGM3 mutation 673:685 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 8 56 theme IgE 1219:1221 arg1 glycosylation 1223:1235 IgE glycosylation 1219:1235 IgE glycosylation 1219:1235 These observations, together with previous knowledge of IgE glycosylation, imply that IgE glycosylation is similarly regulated among healthy control, allergy and PGM3 related hyper IgE syndrome. 26687240 4 57 theme serum 637:641 arg1 1 mL 629:632 1 mL 629:632 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis 629:722 A two-step method was developed to prepare two IgE samples from less than 1 mL of serum collected from a patient with PGM3 mutation and a patient with atopic dermatitis as a control subject. 26687240 0 58 theme IgE 49:51 arg1 syndrome 53:60 a novel hyper IgE syndrome 35:60 a novel hyper IgE syndrome 35:60 Glycoproteomic studies of IgE from a novel hyper IgE syndrome linked to PGM3 mutation. 26687240 1 59 theme antibody 128:135 arg1 activities 137:146 antibody activities 128:146 antibody activities 128:146 Glycans serve as important regulators of antibody activities and half-lives. 26687240 3 60 theme specific 347:354 arg1 glycosylation 360:372 the site specific IgE glycosylation 338:372 the site specific IgE glycosylation 338:372 We therefore describe the site specific IgE glycosylation from a patient with a novel hyper IgE syndrome linked to mutations in PGM3, which is an enzyme involved in synthesizing UDP-GlcNAc, a sugar donor widely required for glycosylation. 26972002 0 0 theme HIV-1 75:79 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 4 1 dep soluble 630:636 arg1 recombinant 639:649 recombinant 639:649 recombinant 639:649 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 0 2 theme Trimeric 66:73 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 2 3 theme enzymes 346:352 arg1 actions 317:323 the actions 313:323 the actions 313:323 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 0 4 theme Glycoprotein 90:101 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 0 4 theme Glycoprotein 90:101 arg1 Sites 55:59 Individual Glycan Sites 37:59 Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein 37:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 4 5 gly glycosylation 602:614 arg1 trimer 651:656 a soluble, recombinant trimer 628:656 a soluble, recombinant trimer (BG505 SOSIP.664) 628:674 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 5 gly glycosylation 602:614 arg1 SOSIP.664 665:673 BG505 SOSIP.664 659:673 BG505 SOSIP.664 659:673 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 1 6 dep glycoprotein 123:134 arg1 trimer 136:141 trimer 136:141 The HIV-1 envelope glycoprotein trimer 104:141 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 4 7 theme processing 732:741 arg1 simplicity 697:706 simplicity 697:706 simplicity 697:706 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 7 theme processing 732:741 arg1 diversity 712:720 diversity 712:720 diversity 712:720 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 0 8 theme Envelope 81:88 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 4 9 from domain 839:844 arg1 clusters 793:800 dense clusters 787:800 dense clusters of oligomannose glycans on the outer domain 787:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 10 theme individual 746:755 arg1 sites 757:761 individual sites 746:761 individual sites 746:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 11 from sites 757:761 arg1 processing 732:741 glycan processing 725:741 glycan processing at individual sites 725:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 11 from sites 757:761 arg1 simplicity 697:706 simplicity 697:706 simplicity 697:706 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 11 from sites 757:761 arg1 diversity 712:720 diversity 712:720 diversity 712:720 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 2 12 theme trimer 262:267 arg1 surface 269:275 the trimer surface 258:275 the trimer surface 258:275 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 2 13 theme glycan-processing 328:344 arg1 enzymes 346:352 glycan-processing enzymes 328:352 glycan-processing enzymes 328:352 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 1 14 link N-linked 169:176 arg1 glycans 178:184 N-linked glycans 169:184 N-linked glycans 169:184 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 2 15 theme under-processed 372:386 arg1 structures 388:397 multiple under-processed structures 363:397 multiple under-processed structures 363:397 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 5 16 dep glycan 926:931 arg1 shield 933:938 shield 933:938 shield 933:938 Although individual sites usually minimally affect the global integrity of the glycan shield, we identify examples of how deleting some glycans can subtly influence neutralization by bNAbs that bind at distant sites. 26972002 3 17 theme oligomannose 431:442 arg1 glycans 444:450 These oligomannose glycans 425:450 These oligomannose glycans 425:450 These oligomannose glycans are recognized by broadly neutralizing antibodies (bNAbs) that are not thwarted by the glycan shield but, paradoxically, target it. 26972002 5 18 theme glycan 926:931 arg1 integrity 909:917 the global integrity 898:917 the global integrity of the glycan shield 898:938 Although individual sites usually minimally affect the global integrity of the glycan shield, we identify examples of how deleting some glycans can subtly influence neutralization by bNAbs that bind at distant sites. 26972002 4 19 theme trimer 651:656 arg1 analysis 616:623 Our site-specific glycosylation analysis 584:623 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) 584:674 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 20 theme glycan 725:730 arg1 processing 732:741 glycan processing 725:741 glycan processing at individual sites 725:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 1 21 theme N-linked 169:176 arg1 glycans 178:184 N-linked glycans 169:184 N-linked glycans 169:184 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 4 22 theme outer 833:837 arg1 domain 839:844 the outer domain 829:844 the outer domain 829:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 2 23 from density 236:242 arg1 surface 269:275 the trimer surface 258:275 the trimer surface 258:275 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 1 24 theme glycans 178:184 arg1 array 160:164 an array 157:164 an array of N-linked glycans that shield it from immune surveillance 157:224 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 2 25 theme steric 285:290 arg1 constraints 292:302 steric constraints 285:302 steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas 285:422 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 4 26 theme soluble 630:636 arg1 trimer 651:656 a soluble, recombinant trimer 628:656 a soluble, recombinant trimer (BG505 SOSIP.664) 628:674 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 26 theme soluble 630:636 arg1 SOSIP.664 665:673 BG505 SOSIP.664 659:673 BG505 SOSIP.664 659:673 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 0 27 theme Antigenic 16:24 arg1 Effects 26:32 Antigenic Effects 16:32 Antigenic Effects 16:32 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 5 28 theme distant 1049:1055 arg1 sites 1057:1061 distant sites 1049:1061 distant sites 1049:1061 Although individual sites usually minimally affect the global integrity of the glycan shield, we identify examples of how deleting some glycans can subtly influence neutralization by bNAbs that bind at distant sites. 26972002 3 29 dep thwarted 523:530 arg1 target 573:578 target 573:578 target it 573:581 These oligomannose glycans are recognized by broadly neutralizing antibodies (bNAbs) that are not thwarted by the glycan shield but, paradoxically, target it. 26972002 3 30 theme neutralizing 478:489 arg1 bNAbs 503:507 bNAbs 503:507 bNAbs 503:507 These oligomannose glycans are recognized by broadly neutralizing antibodies (bNAbs) that are not thwarted by the glycan shield but, paradoxically, target it. 26972002 3 30 theme neutralizing 478:489 arg1 antibodies 491:500 broadly neutralizing antibodies 470:500 broadly neutralizing antibodies (bNAbs) that are not thwarted by the glycan shield but, paradoxically, target it 470:581 These oligomannose glycans are recognized by broadly neutralizing antibodies (bNAbs) that are not thwarted by the glycan shield but, paradoxically, target it. 26972002 0 31 theme Individual 37:46 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 0 31 theme Individual 37:46 arg1 Sites 55:59 Individual Glycan Sites 37:59 Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein 37:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 2 32 theme multiple 363:370 arg1 structures 388:397 multiple under-processed structures 363:397 multiple under-processed structures 363:397 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 2 33 theme specific 409:416 arg1 areas 418:422 specific areas 409:422 specific areas 409:422 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 1 34 gly glycoprotein 123:134 arg1 glycoprotein 123:134 The HIV-1 envelope glycoprotein 104:134 The HIV-1 envelope glycoprotein trimer 104:141 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 4 35 theme clusters 793:800 arg1 mosaic 777:782 a mosaic 775:782 a mosaic of dense clusters of oligomannose glycans on the outer domain 775:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 36 from glycans 818:824 arg1 domain 839:844 the outer domain 829:844 the outer domain 829:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 0 37 theme Sites 55:59 arg1 Composition 0:10 Composition 0:10 Composition 0:10 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 0 37 theme Sites 55:59 arg1 Effects 26:32 Antigenic Effects 16:32 Antigenic Effects 16:32 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 5 38 theme individual 856:865 arg1 sites 867:871 individual sites 856:871 individual sites 856:871 Although individual sites usually minimally affect the global integrity of the glycan shield, we identify examples of how deleting some glycans can subtly influence neutralization by bNAbs that bind at distant sites. 26972002 4 39 theme glycans 818:824 arg1 clusters 793:800 dense clusters 787:800 dense clusters of oligomannose glycans on the outer domain 787:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 1 40 theme HIV-1 108:112 arg1 glycoprotein 123:134 The HIV-1 envelope glycoprotein 104:134 The HIV-1 envelope glycoprotein trimer 104:141 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 0 41 theme Glycan 48:53 arg1 Glycoprotein 90:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 a Trimeric HIV-1 Envelope Glycoprotein 64:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 0 41 theme Glycan 48:53 arg1 Sites 55:59 Individual Glycan Sites 37:59 Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein 37:101 Composition and Antigenic Effects of Individual Glycan Sites of a Trimeric HIV-1 Envelope Glycoprotein. 26972002 4 42 theme oligomannose 805:816 arg1 glycans 818:824 oligomannose glycans 805:824 oligomannose glycans on the outer domain 805:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 1 43 theme envelope 114:121 arg1 glycoprotein 123:134 The HIV-1 envelope glycoprotein 104:134 The HIV-1 envelope glycoprotein trimer 104:141 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 4 44 from diversity 712:720 arg1 sites 757:761 individual sites 746:761 individual sites 746:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 45 theme simplicity 697:706 arg1 extremes 685:692 the extremes 681:692 the extremes of simplicity and diversity of glycan processing at individual sites 681:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 2 46 theme glycans 247:253 arg1 density 236:242 The high density 227:242 The high density of glycans on the trimer surface 227:275 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 4 47 from clusters 793:800 arg1 domain 839:844 the outer domain 829:844 the outer domain 829:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 48 theme diversity 712:720 arg1 extremes 685:692 the extremes 681:692 the extremes of simplicity and diversity of glycan processing at individual sites 681:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 1 49 theme immune 206:211 arg1 surveillance 213:224 immune surveillance 206:224 immune surveillance 206:224 The HIV-1 envelope glycoprotein trimer is covered by an array of N-linked glycans that shield it from immune surveillance. 26972002 5 50 theme global 902:907 arg1 integrity 909:917 the global integrity 898:917 the global integrity of the glycan shield 898:938 Although individual sites usually minimally affect the global integrity of the glycan shield, we identify examples of how deleting some glycans can subtly influence neutralization by bNAbs that bind at distant sites. 26972002 6 51 theme bNAb-targeted 1079:1091 arg1 glycans 1093:1099 bNAb-targeted glycans 1079:1099 bNAb-targeted glycans 1079:1099 The network of bNAb-targeted glycans should be preserved on vaccine antigens. 26972002 4 52 from simplicity 697:706 arg1 sites 757:761 individual sites 746:761 individual sites 746:761 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 53 theme site-specific 588:600 arg1 analysis 616:623 Our site-specific glycosylation analysis 584:623 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) 584:674 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 4 54 theme dense 787:791 arg1 clusters 793:800 dense clusters 787:800 dense clusters of oligomannose glycans on the outer domain 787:844 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 26972002 6 55 theme glycans 1093:1099 arg1 network 1068:1074 The network 1064:1074 The network of bNAb-targeted glycans 1064:1099 The network of bNAb-targeted glycans should be preserved on vaccine antigens. 26972002 6 56 theme vaccine 1124:1130 arg1 antigens 1132:1139 vaccine antigens 1124:1139 vaccine antigens 1124:1139 The network of bNAb-targeted glycans should be preserved on vaccine antigens. 26972002 2 57 theme high 231:234 arg1 density 236:242 The high density 227:242 The high density of glycans on the trimer surface 227:275 The high density of glycans on the trimer surface imposes steric constraints limiting the actions of glycan-processing enzymes, so that multiple under-processed structures remain on specific areas. 26972002 4 58 theme glycosylation 602:614 arg1 analysis 616:623 Our site-specific glycosylation analysis 584:623 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) 584:674 Our site-specific glycosylation analysis of a soluble, recombinant trimer (BG505 SOSIP.664) maps the extremes of simplicity and diversity of glycan processing at individual sites and reveals a mosaic of dense clusters of oligomannose glycans on the outer domain. 25300029 0 0 theme mouse 81:85 arg1 cells 94:98 mouse kidney cells 81:98 mouse kidney cells with the phenotype of polycystic kidney disease 81:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 5 1 theme cyst 1020:1023 arg1 formation 1025:1033 cyst formation 1020:1033 cyst formation 1020:1033 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 2 2 link N-linked 494:501 arg1 glycosylation 503:515 normal and abnormal N-linked glycosylation 474:515 normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 474:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 3 theme Autosomal 149:157 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 3 theme Autosomal 149:157 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 3 theme Autosomal 149:157 arg1 ADPKD 195:199 ADPKD 195:199 ADPKD 195:199 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 4 theme α3 520:521 arg1 subunit 532:538 α3 integrin subunit 520:538 α3 integrin subunit 520:538 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 5 theme normal 307:312 arg1 tissue 321:326 normal kidney tissue 307:326 normal kidney tissue 307:326 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 6 from glycosylation 503:515 arg1 cells 553:557 Pkd1(-/-) cells 543:557 Pkd1(-/-) cells derived from mouse kidneys 543:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 7 theme dominant 159:166 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 7 theme dominant 159:166 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 7 theme dominant 159:166 arg1 ADPKD 195:199 ADPKD 195:199 ADPKD 195:199 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 8 from patterns 462:469 arg1 cells 553:557 Pkd1(-/-) cells 543:557 Pkd1(-/-) cells derived from mouse kidneys 543:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 9 theme glycosylation 503:515 arg1 patterns 462:469 patterns 462:469 patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 462:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 10 theme kidney 314:319 arg1 tissue 321:326 normal kidney tissue 307:326 normal kidney tissue 307:326 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 3 11 theme glycan 611:616 arg1 structures 618:627 Higher molecular weight glycan structures 587:627 Higher molecular weight glycan structures with a different monosaccharide composition 587:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 4 12 located observed 881:888 arg1 cells 910:914 Pkd1(-/-) cells 900:914 Pkd1(-/-) cells 900:914 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 4 12 located observed 881:888 arg1 addition 814:821 addition 814:821 addition 814:821 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 4 12 located observed 881:888 arg2 structure 867:875 an unusual and unique disialic acid glycan structure 824:875 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 0 13 theme kidney 87:92 arg1 cells 94:98 mouse kidney cells 81:98 mouse kidney cells with the phenotype of polycystic kidney disease 81:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 2 14 gly glycosylation 503:515 arg1 subunit 532:538 α3 integrin subunit 520:538 α3 integrin subunit 520:538 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 14 gly glycosylation 503:515 arg1 cells 553:557 Pkd1(-/-) cells 543:557 Pkd1(-/-) cells derived from mouse kidneys 543:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 15 theme abnormal 485:492 arg1 glycosylation 503:515 normal and abnormal N-linked glycosylation 474:515 normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 474:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 0 16 from structures 50:59 arg1 cells 94:98 mouse kidney cells 81:98 mouse kidney cells with the phenotype of polycystic kidney disease 81:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 0 16 from structures 50:59 arg1 integrin 67:74 α3 integrin 64:74 α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 64:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 1 17 theme polycystic 168:177 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 17 theme polycystic 168:177 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 17 theme polycystic 168:177 arg1 ADPKD 195:199 ADPKD 195:199 ADPKD 195:199 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 18 theme nano-HPLC-mass 408:421 arg1 approach 437:444 a powerful nano-HPLC-mass spectrometric approach 397:444 a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 397:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 5 19 contain have 985:988 arg2 role 992:995 a role 990:995 a role 990:995 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 5 19 contain have 985:988 arg1 glycosylation 967:979 abnormal protein glycosylation 950:979 abnormal protein glycosylation 950:979 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 4 20 theme acid 855:858 arg1 structure 867:875 an unusual and unique disialic acid glycan structure 824:875 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 1 21 theme kidney 179:184 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 21 theme kidney 179:184 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 21 theme kidney 179:184 arg1 ADPKD 195:199 ADPKD 195:199 ADPKD 195:199 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 22 theme powerful 399:406 arg1 approach 437:444 a powerful nano-HPLC-mass spectrometric approach 397:444 a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 397:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 4 23 theme Pkd1 900:903 arg1 cells 910:914 Pkd1(-/-) cells 900:914 Pkd1(-/-) cells 900:914 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 2 24 used utilized 388:395 arg2 we 380:381 we 380:381 we 380:381 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 25 dep Pkd1 258:261 arg1 genes 271:275 genes 271:275 genes 271:275 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 26 from cells 553:557 arg1 patterns 462:469 patterns 462:469 patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 462:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 27 theme mouse 572:576 arg1 kidneys 578:584 mouse kidneys 572:584 mouse kidneys 572:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 0 28 with cells 94:98 arg1 phenotype 109:117 the phenotype 105:117 the phenotype of polycystic kidney disease 105:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 3 29 attach isolated 750:757 arg2 integrin 741:748 α3 integrin 738:748 α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells 738:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 29 attach isolated 750:757 arg1 cells 774:778 Pkd1(+/+) cells 764:778 Pkd1(+/+) cells compared with Pkd1(-/-) cells 764:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 5 30 theme protein 959:965 arg1 glycosylation 967:979 abnormal protein glycosylation 950:979 abnormal protein glycosylation 950:979 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 0 31 theme glycans 24:30 arg1 Identification 0:13 Identification 0:13 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 0:147 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 2 32 theme subunit 532:538 arg1 glycosylation 503:515 normal and abnormal N-linked glycosylation 474:515 normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 474:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 0 33 theme novel 18:22 arg1 glycans 24:30 novel glycans 18:30 novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 18:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 2 34 theme integrin 523:530 arg1 subunit 532:538 α3 integrin subunit 520:538 α3 integrin subunit 520:538 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 1 35 theme end-stage 340:348 arg1 disease 357:363 end-stage kidney disease 340:363 end-stage kidney disease 340:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 0 36 theme disialylated 37:48 arg1 structures 50:59 disialylated structures 37:59 disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 37:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 0 37 theme kidney 133:138 arg1 disease 140:146 polycystic kidney disease 122:146 polycystic kidney disease 122:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 1 38 theme kidney 350:355 arg1 disease 357:363 end-stage kidney disease 340:363 end-stage kidney disease 340:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 4 39 theme disialic 846:853 arg1 acid 855:858 an unusual and unique disialic acid 824:858 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 3 40 with structures 618:627 arg1 composition 661:671 a different monosaccharide composition 634:671 a different monosaccharide composition 634:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 4 41 theme -/- 905:907 arg1 cells 910:914 Pkd1(-/-) cells 900:914 Pkd1(-/-) cells 900:914 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 3 42 theme α3 738:739 arg1 integrin 741:748 α3 integrin 738:748 α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells 738:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 0 43 theme polycystic 122:131 arg1 disease 140:146 polycystic kidney disease 122:146 polycystic kidney disease 122:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 3 44 located observed 678:685 arg1 sites 694:698 two sites 690:698 two sites 690:698 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 44 located observed 678:685 arg2 structures 618:627 Higher molecular weight glycan structures 587:627 Higher molecular weight glycan structures with a different monosaccharide composition 587:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 44 located observed 678:685 arg1 sites 729:733 Asn-925 and Asn-928 sites 709:733 Asn-925 and Asn-928 sites 709:733 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 44 located observed 678:685 arg1 integrin 741:748 α3 integrin 738:748 α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells 738:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 0 45 with glycans 24:30 arg1 structures 50:59 disialylated structures 37:59 disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 37:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 2 46 theme spectrometric 423:435 arg1 approach 437:444 a powerful nano-HPLC-mass spectrometric approach 397:444 a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 397:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 3 47 theme weight 604:609 arg1 structures 618:627 Higher molecular weight glycan structures 587:627 Higher molecular weight glycan structures with a different monosaccharide composition 587:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 48 theme Higher 587:592 arg1 structures 618:627 Higher molecular weight glycan structures 587:627 Higher molecular weight glycan structures with a different monosaccharide composition 587:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 4 49 theme unusual 827:833 arg1 acid 855:858 an unusual and unique disialic acid 824:858 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 5 50 theme formation 1025:1033 arg1 pathogenesis 1004:1015 the pathogenesis 1000:1015 the pathogenesis of cyst formation in ADPKD 1000:1042 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 1 51 from mutations 241:249 arg1 Pkd2 266:269 Pkd2 266:269 Pkd2 266:269 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 51 from mutations 241:249 arg1 Pkd1 258:261 the Pkd1 254:261 the Pkd1 254:261 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 0 52 theme disease 140:146 arg1 phenotype 109:117 the phenotype 105:117 the phenotype of polycystic kidney disease 105:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 3 53 theme different 636:644 arg1 composition 661:671 a different monosaccharide composition 634:671 a different monosaccharide composition 634:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 54 theme molecular 594:602 arg1 structures 618:627 Higher molecular weight glycan structures 587:627 Higher molecular weight glycan structures with a different monosaccharide composition 587:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 1 55 theme large 287:291 arg1 cysts 293:297 large cysts 287:297 large cysts 287:297 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 56 theme Pkd1 543:546 arg1 cells 553:557 Pkd1(-/-) cells 543:557 Pkd1(-/-) cells derived from mouse kidneys 543:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 3 57 theme monosaccharide 646:659 arg1 composition 661:671 a different monosaccharide composition 634:671 a different monosaccharide composition 634:671 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 3 58 theme -/- 799:801 arg1 cells 804:808 Pkd1(-/-) cells 794:808 Pkd1(-/-) cells 794:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 1 59 theme common 207:212 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 59 theme common 207:212 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 0 60 from cells 94:98 arg1 structures 50:59 disialylated structures 37:59 disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 37:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 0 60 from cells 94:98 arg1 integrin 67:74 α3 integrin 64:74 α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 64:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 0 61 gly disialylated 37:48 arg1 structures 50:59 disialylated structures 37:59 disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 37:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 4 62 theme glycan 860:865 arg1 structure 867:875 an unusual and unique disialic acid glycan structure 824:875 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 5 63 theme abnormal 950:957 arg1 glycosylation 967:979 abnormal protein glycosylation 950:979 abnormal protein glycosylation 950:979 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 1 64 theme genetic 214:220 arg1 disorder 222:229 a common genetic disorder 205:229 a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease 205:363 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 1 64 theme genetic 214:220 arg1 disease 186:192 Autosomal dominant polycystic kidney disease 149:192 Autosomal dominant polycystic kidney disease (ADPKD) 149:200 Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder caused by mutations in the Pkd1 or Pkd2 genes, in which large cysts replace normal kidney tissue, leading to end-stage kidney disease. 25300029 2 65 theme normal 474:479 arg1 glycosylation 503:515 normal and abnormal N-linked glycosylation 474:515 normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 474:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 4 66 theme unique 839:844 arg1 acid 855:858 an unusual and unique disialic acid 824:858 an unusual and unique disialic acid glycan structure 824:875 In addition, an unusual and unique disialic acid glycan structure was observed solely in Pkd1(-/-) cells. 25300029 5 67 from pathogenesis 1004:1015 arg1 ADPKD 1038:1042 ADPKD 1038:1042 ADPKD 1038:1042 Thus, these studies suggest that abnormal protein glycosylation may have a role on the pathogenesis of cyst formation in ADPKD. 25300029 3 68 theme Pkd1 794:797 arg1 cells 804:808 Pkd1(-/-) cells 794:808 Pkd1(-/-) cells 794:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 0 69 theme α3 64:65 arg1 integrin 67:74 α3 integrin 64:74 α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease 64:146 Identification of novel glycans with disialylated structures in α3 integrin from mouse kidney cells with the phenotype of polycystic kidney disease. 25300029 3 70 theme Pkd1 764:767 arg1 cells 774:778 Pkd1(+/+) cells 764:778 Pkd1(+/+) cells compared with Pkd1(-/-) cells 764:808 Higher molecular weight glycan structures with a different monosaccharide composition were observed at two sites, namely, Asn-925 and Asn-928 sites in α3 integrin isolated from Pkd1(+/+) cells compared with Pkd1(-/-) cells. 25300029 2 71 attach derived 559:565 arg1 kidneys 578:584 mouse kidneys 572:584 mouse kidneys 572:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 71 attach derived 559:565 arg2 cells 553:557 Pkd1(-/-) cells 543:557 Pkd1(-/-) cells derived from mouse kidneys 543:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 25300029 2 72 theme N-linked 494:501 arg1 glycosylation 503:515 normal and abnormal N-linked glycosylation 474:515 normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys 474:584 In this study we have utilized a powerful nano-HPLC-mass spectrometric approach to characterize patterns of normal and abnormal N-linked glycosylation of α3 integrin subunit in Pkd1(-/-) cells derived from mouse kidneys. 24803430 3 0 from O-fucose 645:652 arg1 EGF12 665:669 EGF12 665:669 EGF12 665:669 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 1 theme Notch1 486:491 arg1 hN1 494:496 hN1 494:496 hN1 494:496 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 1 theme Notch1 486:491 arg1 region 450:455 the ligand-binding region 431:455 the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) 431:497 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 1 theme Notch1 486:491 arg1 domains 462:468 EGF domains 11-13 458:474 EGF domains 11-13 458:474 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 1 2 theme homeostasis 240:250 arg1 aspects 180:186 many aspects 175:186 many aspects of development, cell fate determination, and tissue homeostasis 175:250 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 0 3 theme Notch1 77:82 arg1 region 67:72 the ligand-binding region 48:72 the ligand-binding region of Notch1 48:82 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 0 4 from extension 16:24 arg1 region 67:72 the ligand-binding region 48:72 the ligand-binding region of Notch1 48:82 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 4 5 theme hN1 826:828 arg1 structures 791:800 the crystal structures 779:800 the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide 779:911 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 0 6 link O-linked 29:36 arg1 fucose 38:43 O-linked fucose 29:43 O-linked fucose 29:43 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 6 7 theme Fringe-catalyzed 1222:1237 arg1 enhancement 1239:1249 the Fringe-catalyzed enhancement 1218:1249 the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans 1218:1303 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 6 8 theme Notch 1418:1422 arg1 regions 1407:1413 other regions 1401:1413 other regions of Notch 1401:1422 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 2 9 theme Notch 328:332 arg1 receptor 334:341 the Notch receptor 324:341 the Notch receptor 324:341 Notch signaling can be modulated by posttranslational modifications to the Notch receptor, which are known to alter both ligand binding and receptor activation. 24803430 0 10 theme Notch 115:119 arg1 ligands 121:127 mammalian Notch ligands 105:127 mammalian Notch ligands 105:127 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 4 11 theme domains 809:815 arg1 structures 791:800 the crystal structures 779:800 the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide 779:911 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 0 12 theme mammalian 105:113 arg1 ligands 121:127 mammalian Notch ligands 105:127 mammalian Notch ligands 105:127 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 5 13 theme ligand-binding 1066:1079 arg1 site 1081:1084 the ligand-binding site 1062:1084 the ligand-binding site in hN1 EGF12 1062:1097 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 4 14 theme EGF 805:807 arg1 domains 809:815 EGF domains 11-13 805:821 EGF domains 11-13 805:821 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 3 15 dep O-fucose 504:511 arg1 glycans 527:533 glycans 527:533 glycans 527:533 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 1 16 theme many 175:178 arg1 aspects 180:186 many aspects 175:186 many aspects of development, cell fate determination, and tissue homeostasis 175:250 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 5 17 theme region 1143:1148 arg1 affinity 1126:1133 an increased affinity 1113:1133 an increased affinity of this region for ligands Jagged1 and DLL1 1113:1177 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 2 18 theme ligand 374:379 arg1 binding 381:387 ligand binding 374:387 ligand binding 374:387 Notch signaling can be modulated by posttranslational modifications to the Notch receptor, which are known to alter both ligand binding and receptor activation. 24803430 0 19 theme Fringe-mediated 0:14 arg1 extension 16:24 Fringe-mediated extension 0:24 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 0:82 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 3 20 theme 1 729:729 arg1 ligands 738:744 Delta-like 1 (DLL1) ligands 718:744 Delta-like 1 (DLL1) ligands 718:744 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 5 21 dep ligands 1154:1160 arg1 Jagged1 1162:1168 Jagged1 1162:1168 Jagged1 1162:1168 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 5 21 dep ligands 1154:1160 arg1 ligands 1154:1160 ligands Jagged1 and DLL1 1154:1177 ligands Jagged1 and DLL1 1154:1177 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 5 21 dep ligands 1154:1160 arg1 DLL1 1174:1177 DLL1 1174:1177 DLL1 1174:1177 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 0 22 theme O-linked 29:36 arg1 fucose 38:43 O-linked fucose 29:43 O-linked fucose 29:43 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 4 23 theme GlcNAc-fucose 886:898 arg1 disaccharide 900:911 the GlcNAc-fucose disaccharide 882:911 the GlcNAc-fucose disaccharide 882:911 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 6 24 theme inhibitory 1327:1336 arg1 effect 1338:1343 the inhibitory effect 1323:1343 the inhibitory effect of Fringe on Jagged/Serrate mediated signaling 1323:1390 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 1 25 theme development 191:201 arg1 aspects 180:186 many aspects 175:186 many aspects of development, cell fate determination, and tissue homeostasis 175:250 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 6 26 theme other 1401:1405 arg1 regions 1407:1413 other regions 1401:1413 other regions of Notch 1401:1422 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 6 27 theme mediated 1373:1380 arg1 signaling 1382:1390 Jagged/Serrate mediated signaling 1358:1390 Jagged/Serrate mediated signaling 1358:1390 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 2 28 theme posttranslational 289:305 arg1 modifications 307:319 posttranslational modifications 289:319 posttranslational modifications 289:319 Notch signaling can be modulated by posttranslational modifications to the Notch receptor, which are known to alter both ligand binding and receptor activation. 24803430 3 29 from T466 657:660 arg1 O-fucose 645:652 the O-fucose 641:652 the O-fucose at T466 in EGF12 641:669 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 30 theme Delta-like 718:727 arg1 ligands 738:744 Delta-like 1 (DLL1) ligands 718:744 Delta-like 1 (DLL1) ligands 718:744 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 31 theme GlcNAc 631:636 arg1 addition 619:626 the Fringe-catalyzed addition 598:626 the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 598:669 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 4 32 theme crystal 783:789 arg1 structures 791:800 the crystal structures 779:800 the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide 779:911 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 1 33 theme cell 204:207 arg1 determination 214:226 cell fate determination 204:226 cell fate determination 204:226 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 0 34 theme fucose 38:43 arg1 extension 16:24 Fringe-mediated extension 0:24 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 0:82 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 4 35 theme backbone 956:963 arg1 structure 965:973 backbone structure 956:973 backbone structure 956:973 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 3 36 theme Fringe-catalyzed 602:617 arg1 addition 619:626 the Fringe-catalyzed addition 598:626 the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 598:669 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 37 theme surface 567:573 arg1 resonance 583:591 surface plasmon resonance 567:591 surface plasmon resonance 567:591 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 2 38 theme receptor 393:400 arg1 activation 402:411 receptor activation 393:411 receptor activation 393:411 Notch signaling can be modulated by posttranslational modifications to the Notch receptor, which are known to alter both ligand binding and receptor activation. 24803430 0 39 theme ligand-binding 52:65 arg1 region 67:72 the ligand-binding region 48:72 the ligand-binding region of Notch1 48:82 Fringe-mediated extension of O-linked fucose in the ligand-binding region of Notch1 increases binding to mammalian Notch ligands. 24803430 4 40 theme O-fucose 855:862 arg1 monosaccharide 864:877 the O-fucose monosaccharide 851:877 the O-fucose monosaccharide 851:877 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 3 41 theme flow 548:551 arg1 cytometry 553:561 flow cytometry 548:561 flow cytometry 548:561 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 5 42 from site 1081:1084 arg1 hN1 1089:1091 hN1 EGF12 1089:1097 hN1 EGF12 1089:1097 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 3 43 theme human 480:484 arg1 Notch1 486:491 human Notch1 480:491 human Notch1 480:491 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 6 44 located observed 1276:1283 arg2 enhancement 1239:1249 the Fringe-catalyzed enhancement 1218:1249 the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans 1218:1303 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 6 44 located observed 1276:1283 arg1 humans 1298:1303 humans 1298:1303 humans 1298:1303 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 6 44 located observed 1276:1283 arg1 flies 1288:1292 flies 1288:1292 flies 1288:1292 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 5 45 theme increased 1116:1124 arg1 affinity 1126:1133 an increased affinity 1113:1133 an increased affinity of this region for ligands Jagged1 and DLL1 1113:1177 Collectively, these data demonstrate a role for GlcNAc in modulating the ligand-binding site in hN1 EGF12, resulting in an increased affinity of this region for ligands Jagged1 and DLL1. 24803430 6 46 theme signaling 1266:1274 arg1 enhancement 1239:1249 the Fringe-catalyzed enhancement 1218:1249 the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans 1218:1303 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 1 47 theme determination 214:226 arg1 aspects 180:186 many aspects 175:186 many aspects of development, cell fate determination, and tissue homeostasis 175:250 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 6 48 theme Notch-Delta 1254:1264 arg1 signaling 1266:1274 Notch-Delta signaling 1254:1274 Notch-Delta signaling 1254:1274 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 2 49 theme Notch 253:257 arg1 signaling 259:267 Notch signaling 253:267 Notch signaling 253:267 Notch signaling can be modulated by posttranslational modifications to the Notch receptor, which are known to alter both ligand binding and receptor activation. 24803430 1 50 theme Notch 134:138 arg1 pathway 150:156 The Notch signaling pathway 130:156 The Notch signaling pathway 130:156 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 1 50 theme Notch 134:138 arg1 essential 161:169 essential 161:169 essential 161:169 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 4 51 from change 946:951 arg1 structure 965:973 backbone structure 956:973 backbone structure 956:973 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 6 52 theme Fringe 1348:1353 arg1 effect 1338:1343 the inhibitory effect 1323:1343 the inhibitory effect of Fringe on Jagged/Serrate mediated signaling 1323:1390 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 1 53 theme fate 209:212 arg1 determination 214:226 cell fate determination 204:226 cell fate determination 204:226 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 3 54 theme plasmon 575:581 arg1 resonance 583:591 surface plasmon resonance 567:591 surface plasmon resonance 567:591 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 55 theme EGF 458:460 arg1 region 450:455 the ligand-binding region 431:455 the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) 431:497 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 55 theme EGF 458:460 arg1 domains 462:468 EGF domains 11-13 458:474 EGF domains 11-13 458:474 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 1 56 theme signaling 140:148 arg1 pathway 150:156 The Notch signaling pathway 130:156 The Notch signaling pathway 130:156 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 1 56 theme signaling 140:148 arg1 essential 161:169 essential 161:169 essential 161:169 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 24803430 3 57 theme ligand-binding 435:448 arg1 hN1 494:496 hN1 494:496 hN1 494:496 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 57 theme ligand-binding 435:448 arg1 region 450:455 the ligand-binding region 431:455 the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) 431:497 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 3 57 theme ligand-binding 435:448 arg1 domains 462:468 EGF domains 11-13 458:474 EGF domains 11-13 458:474 We have modified the ligand-binding region (EGF domains 11-13) of human Notch1 (hN1) with O-fucose and O-glucose glycans and shown by flow cytometry and surface plasmon resonance that the Fringe-catalyzed addition of GlcNAc to the O-fucose at T466 in EGF12 substantially increases binding to Jagged1 and Delta-like 1 (DLL1) ligands. 24803430 6 58 from effect 1338:1343 arg1 signaling 1382:1390 Jagged/Serrate mediated signaling 1358:1390 Jagged/Serrate mediated signaling 1358:1390 We propose that this finding explains the Fringe-catalyzed enhancement of Notch-Delta signaling observed in flies and humans, but suggest that the inhibitory effect of Fringe on Jagged/Serrate mediated signaling involves other regions of Notch. 24803430 4 59 theme EGF12 924:928 arg1 T466 916:919 T466 916:919 T466 of EGF12 916:928 We have subsequently determined the crystal structures of EGF domains 11-13 of hN1 modified with either the O-fucose monosaccharide or the GlcNAc-fucose disaccharide at T466 of EGF12 and observed no change in backbone structure for each variant. 24803430 1 60 theme tissue 233:238 arg1 homeostasis 240:250 tissue homeostasis 233:250 tissue homeostasis 233:250 The Notch signaling pathway is essential for many aspects of development, cell fate determination, and tissue homeostasis. 26868756 6 0 used used 892:895 arg2 GC 873:874 The recombinant GC 857:874 The recombinant GC 857:874 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 4 1 theme promising 665:673 arg1 system 675:680 a promising system 663:680 a promising system for the production of therapeutic glycoproteins in plants 663:738 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 4 1 theme promising 665:673 arg1 NbGNTI-RNAi 642:652 this NbGNTI-RNAi 637:652 this NbGNTI-RNAi 637:652 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 7 2 theme GC 1129:1130 arg1 sites 1108:1112 the four occupied N-glycosylation sites 1074:1112 the four occupied N-glycosylation sites of recombinant GC 1074:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 2 theme GC 1129:1130 arg1 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 6 3 theme therapeutic 998:1008 arg1 efficacy 1010:1017 its therapeutic efficacy 994:1017 its therapeutic efficacy 994:1017 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 6 4 from therapy 920:926 arg1 patients 931:938 patients 931:938 patients with Gaucher's disease 931:961 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 0 5 from production 4:13 arg1 plants 85:90 glyco-engineered Nicotiana benthamiana plants 46:90 glyco-engineered Nicotiana benthamiana plants 46:90 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 1 6 theme β1,2-xylose 163:173 arg1 presence 151:158 the presence 147:158 the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures 147:225 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 1 7 theme antigenic 254:262 arg1 activity 264:271 their antigenic activity 248:271 their antigenic activity 248:271 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 4 8 gly glycoproteins 716:728 arg1 glycoproteins 716:728 therapeutic glycoproteins 704:728 therapeutic glycoproteins 704:728 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 6 9 theme enzyme 901:906 arg1 therapy 920:926 enzyme replacement therapy 901:926 enzyme replacement therapy in patients with Gaucher's disease 901:961 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 7 10 theme β1,2-xylose 1299:1309 arg1 epitopes 1327:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 5 11 theme transgenic 789:798 arg1 N. benthamiana 800:813 transgenic N. benthamiana 789:813 transgenic N. benthamiana expressing human glucocerebrosidase (GC) 789:854 The NbGNTI-RNAi plant was cross-pollinated with transgenic N. benthamiana expressing human glucocerebrosidase (GC). 26868756 2 12 theme RNA 289:291 arg1 interference 293:304 RNA interference 289:304 RNA interference (RNAi) technology 289:322 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 2 12 theme RNA 289:291 arg1 RNAi 307:310 RNAi 307:310 RNAi 307:310 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 7 13 theme recombinant 1117:1127 arg1 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 3 14 theme strong 492:497 arg1 reduction 499:507 a strong reduction 490:507 a strong reduction of plant-specific N-glycans 490:535 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 7 15 gly N-glycosylation 1092:1106 arg1 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 15 gly N-glycosylation 1092:1106 arg2 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 15 gly N-glycosylation 1092:1106 arg2 sites 1108:1112 the four occupied N-glycosylation sites 1074:1112 the four occupied N-glycosylation sites of recombinant GC 1074:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 15 gly N-glycosylation 1092:1106 arg2 four 1078:1081 four 1078:1081 four 1078:1081 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 10 16 theme GC 1735:1736 arg1 production 1721:1730 the production 1717:1730 the production of GC with customized N-glycan structures 1717:1772 Therefore, it would provide a robust tool for the production of GC with customized N-glycan structures. 26868756 2 17 theme GNTI 400:403 arg1 expression 406:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression 350:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana 350:440 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 7 18 theme NbGNTI-RNAi 1135:1145 arg1 GC 1155:1156 GC 1155:1156 GC(gnt1) 1155:1162 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 18 theme NbGNTI-RNAi 1135:1145 arg1 plants 1147:1152 NbGNTI-RNAi plants 1135:1152 NbGNTI-RNAi plants (GC(gnt1) ) 1135:1164 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 6 19 theme recombinant 861:871 arg1 GC 873:874 The recombinant GC 857:874 The recombinant GC 857:874 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 1 20 theme core 179:182 arg1 α1,3-fucose 184:194 core α1,3-fucose 179:194 core α1,3-fucose 179:194 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 7 21 dep % 1209:1209 arg1 to 1214:1215 to 1214:1215 to 1214:1215 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 21 dep % 1209:1209 arg1 up 1211:1212 up 1211:1212 up 1211:1212 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 1 22 theme α1,3-fucose 184:194 arg1 presence 151:158 the presence 147:158 the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures 147:225 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 5 23 theme human 826:830 arg1 GC 852:853 GC 852:853 GC 852:853 The NbGNTI-RNAi plant was cross-pollinated with transgenic N. benthamiana expressing human glucocerebrosidase (GC). 26868756 5 23 theme human 826:830 arg1 glucocerebrosidase 832:849 human glucocerebrosidase 826:849 human glucocerebrosidase (GC) 826:854 The NbGNTI-RNAi plant was cross-pollinated with transgenic N. benthamiana expressing human glucocerebrosidase (GC). 26868756 6 24 theme terminal 973:980 arg1 mannose 982:988 terminal mannose 973:988 terminal mannose for its therapeutic efficacy 973:1017 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 7 25 theme occupied 1083:1090 arg1 sites 1108:1112 the four occupied N-glycosylation sites 1074:1112 the four occupied N-glycosylation sites of recombinant GC 1074:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 25 theme occupied 1083:1090 arg1 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 4 26 theme glycoproteins 716:728 arg1 production 690:699 the production 686:699 the production of therapeutic glycoproteins in plants 686:738 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 2 27 theme N-acetylglucosaminyltransferase 365:395 arg1 expression 406:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression 350:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana 350:440 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 0 28 theme human 18:22 arg1 glucocerebrosidase 24:41 human glucocerebrosidase 18:41 human glucocerebrosidase 18:41 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 2 29 used used 328:331 arg2 technology 313:322 RNA interference (RNAi) technology 289:322 RNA interference (RNAi) technology 289:322 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 2 30 theme interference 293:304 arg1 technology 313:322 RNA interference (RNAi) technology 289:322 RNA interference (RNAi) technology 289:322 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 10 31 theme robust 1701:1706 arg1 tool 1708:1711 a robust tool 1699:1711 a robust tool for the production of GC with customized N-glycan structures 1699:1772 Therefore, it would provide a robust tool for the production of GC with customized N-glycan structures. 26868756 2 32 theme Nicotiana 420:428 arg1 benthamiana 430:440 Nicotiana benthamiana 420:440 Nicotiana benthamiana 420:440 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 6 33 with patients 931:938 arg1 disease 955:961 Gaucher's disease 945:961 Gaucher's disease 945:961 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 10 34 theme customized 1743:1752 arg1 structures 1763:1772 customized N-glycan structures 1743:1772 customized N-glycan structures 1743:1772 Therefore, it would provide a robust tool for the production of GC with customized N-glycan structures. 26868756 9 35 theme NbGNTI-RNAi 1600:1610 arg1 plants 1612:1617 the NbGNTI-RNAi plants 1596:1617 the NbGNTI-RNAi plants 1596:1617 Notably, the NbGNTI-RNAi line, producing GC, was stable and the NbGNTI-RNAi plants were viable and did not show any obvious phenotype. 26868756 9 35 theme NbGNTI-RNAi 1600:1610 arg1 viable 1624:1629 viable 1624:1629 viable 1624:1629 Notably, the NbGNTI-RNAi line, producing GC, was stable and the NbGNTI-RNAi plants were viable and did not show any obvious phenotype. 26868756 7 36 contain had 1241:1243 arg1 majority 1182:1189 the majority 1178:1189 the majority (ranging from 73.3% up to 85.5%) of the N-glycans 1178:1239 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 36 contain had 1241:1243 arg2 structures 1258:1267 mannose-type structures 1245:1267 mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1245:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 1 37 theme therapeutic 115:125 arg1 proteins 127:134 therapeutic proteins 115:134 therapeutic proteins 115:134 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 3 38 theme N-glycans 527:535 arg1 reduction 499:507 a strong reduction 490:507 a strong reduction of plant-specific N-glycans 490:535 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 8 39 theme target 1483:1488 arg1 liver 1461:1465 liver 1461:1465 liver 1461:1465 Moreover, GC(gnt1) could be taken up into the macrophage cells via mannose receptors, and distributed and taken up into the liver and spleen, the target organs in the treatment of Gaucher's disease. 26868756 8 39 theme target 1483:1488 arg1 organs 1490:1495 the target organs 1479:1495 the target organs in the treatment of Gaucher's disease 1479:1533 Moreover, GC(gnt1) could be taken up into the macrophage cells via mannose receptors, and distributed and taken up into the liver and spleen, the target organs in the treatment of Gaucher's disease. 26868756 7 40 theme N-glycans 1231:1239 arg1 majority 1182:1189 the majority 1178:1189 the majority (ranging from 73.3% up to 85.5%) of the N-glycans 1178:1239 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 8 41 from organs 1490:1495 arg1 treatment 1504:1512 the treatment 1500:1512 the treatment of Gaucher's disease 1500:1533 Moreover, GC(gnt1) could be taken up into the macrophage cells via mannose receptors, and distributed and taken up into the liver and spleen, the target organs in the treatment of Gaucher's disease. 26868756 7 42 theme potential 1277:1285 arg1 epitopes 1327:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 1 43 theme N-glycan 207:214 arg1 structures 216:225 plants' N-glycan structures 199:225 plants' N-glycan structures 199:225 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 0 44 theme glucocerebrosidase 24:41 arg1 production 4:13 The production 0:13 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants 0:90 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 7 45 theme immunogenic 1287:1297 arg1 epitopes 1327:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 3 46 theme total 583:587 arg1 N-glycans 589:597 the total N-glycans 579:597 the total N-glycans 579:597 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 0 47 theme Nicotiana 63:71 arg1 plants 85:90 glyco-engineered Nicotiana benthamiana plants 46:90 glyco-engineered Nicotiana benthamiana plants 46:90 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 5 48 theme NbGNTI-RNAi 745:755 arg1 plant 757:761 The NbGNTI-RNAi plant 741:761 The NbGNTI-RNAi plant 741:761 The NbGNTI-RNAi plant was cross-pollinated with transgenic N. benthamiana expressing human glucocerebrosidase (GC). 26868756 7 49 theme N-glycan 1024:1031 arg1 structures 1033:1042 The N-glycan structures 1020:1042 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) 1020:1164 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 1 50 theme proteins 127:134 arg1 production 101:110 the production 97:110 the production of therapeutic proteins in plants 97:144 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 0 51 theme glyco-engineered 46:61 arg1 plants 85:90 glyco-engineered Nicotiana benthamiana plants 46:90 glyco-engineered Nicotiana benthamiana plants 46:90 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 10 52 with production 1721:1730 arg1 structures 1763:1772 customized N-glycan structures 1743:1772 customized N-glycan structures 1743:1772 Therefore, it would provide a robust tool for the production of GC with customized N-glycan structures. 26868756 6 53 theme replacement 908:918 arg1 therapy 920:926 enzyme replacement therapy 901:926 enzyme replacement therapy in patients with Gaucher's disease 901:961 The recombinant GC, which has been used for enzyme replacement therapy in patients with Gaucher's disease, requires terminal mannose for its therapeutic efficacy. 26868756 3 54 theme plant-specific 512:525 arg1 N-glycans 527:535 plant-specific N-glycans 512:535 plant-specific N-glycans 512:535 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 8 55 theme mannose 1404:1410 arg1 receptors 1412:1420 mannose receptors 1404:1420 mannose receptors 1404:1420 Moreover, GC(gnt1) could be taken up into the macrophage cells via mannose receptors, and distributed and taken up into the liver and spleen, the target organs in the treatment of Gaucher's disease. 26868756 8 56 theme macrophage 1383:1392 arg1 cells 1394:1398 the macrophage cells 1379:1398 the macrophage cells 1379:1398 Moreover, GC(gnt1) could be taken up into the macrophage cells via mannose receptors, and distributed and taken up into the liver and spleen, the target organs in the treatment of Gaucher's disease. 26868756 9 57 theme NbGNTI-RNAi 1549:1559 arg1 line 1561:1564 the NbGNTI-RNAi line 1545:1564 the NbGNTI-RNAi line 1545:1564 Notably, the NbGNTI-RNAi line, producing GC, was stable and the NbGNTI-RNAi plants were viable and did not show any obvious phenotype. 26868756 9 57 theme NbGNTI-RNAi 1549:1559 arg1 stable 1585:1590 stable 1585:1590 stable 1585:1590 Notably, the NbGNTI-RNAi line, producing GC, was stable and the NbGNTI-RNAi plants were viable and did not show any obvious phenotype. 26868756 7 58 theme N-glycosylation 1092:1106 arg1 sites 1108:1112 the four occupied N-glycosylation sites 1074:1112 the four occupied N-glycosylation sites of recombinant GC 1074:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 58 theme N-glycosylation 1092:1106 arg1 GC 1129:1130 recombinant GC 1117:1130 recombinant GC 1117:1130 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 4 59 theme therapeutic 704:714 arg1 glycoproteins 716:728 therapeutic glycoproteins 704:728 therapeutic glycoproteins 704:728 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 9 60 theme obvious 1652:1658 arg1 phenotype 1660:1668 any obvious phenotype 1648:1668 any obvious phenotype 1648:1668 Notably, the NbGNTI-RNAi line, producing GC, was stable and the NbGNTI-RNAi plants were viable and did not show any obvious phenotype. 26868756 3 61 theme high-mannose 607:618 arg1 type 620:623 high-mannose type 607:623 high-mannose type 607:623 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 2 62 from expression 406:415 arg1 benthamiana 430:440 Nicotiana benthamiana 420:440 Nicotiana benthamiana 420:440 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 4 63 from production 690:699 arg1 plants 733:738 plants 733:738 plants 733:738 Therefore, this NbGNTI-RNAi would be a promising system for the production of therapeutic glycoproteins in plants. 26868756 3 64 theme glyco-engineered 447:462 arg1 NbGNTI-RNAi 470:480 NbGNTI-RNAi 470:480 NbGNTI-RNAi 470:480 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 3 64 theme glyco-engineered 447:462 arg1 line 464:467 One glyco-engineered line 443:467 One glyco-engineered line (NbGNTI-RNAi) 443:481 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 7 65 theme 73.3 1205:1208 arg1 % 1209:1209 % 1209:1209 % 1209:1209 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 10 66 theme N-glycan 1754:1761 arg1 structures 1763:1772 customized N-glycan structures 1743:1772 customized N-glycan structures 1743:1772 Therefore, it would provide a robust tool for the production of GC with customized N-glycan structures. 26868756 3 67 theme N-glycans 589:597 arg1 % 574:574 as much as 90.9% 559:574 as much as 90.9% of the total N-glycans 559:597 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 3 67 theme N-glycans 589:597 arg1 N-glycans 589:597 the total N-glycans 579:597 the total N-glycans 579:597 One glyco-engineered line (NbGNTI-RNAi) showed a strong reduction of plant-specific N-glycans, with the result that as much as 90.9% of the total N-glycans were of high-mannose type. 26868756 7 68 theme mannose-type 1245:1256 arg1 structures 1258:1267 mannose-type structures 1245:1267 mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1245:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 2 69 theme endogenous 354:363 arg1 expression 406:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression 350:415 the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana 350:440 In this study, RNA interference (RNAi) technology was used to down-regulate the endogenous N-acetylglucosaminyltransferase I (GNTI) expression in Nicotiana benthamiana. 26868756 1 70 from production 101:110 arg1 plants 139:144 plants 139:144 plants 139:144 For the production of therapeutic proteins in plants, the presence of β1,2-xylose and core α1,3-fucose on plants' N-glycan structures has been debated for their antigenic activity. 26868756 0 71 theme benthamiana 73:83 arg1 plants 85:90 glyco-engineered Nicotiana benthamiana plants 46:90 glyco-engineered Nicotiana benthamiana plants 46:90 The production of human glucocerebrosidase in glyco-engineered Nicotiana benthamiana plants. 26868756 7 72 theme α1,3-fucose 1315:1325 arg1 epitopes 1327:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 potential immunogenic β1,2-xylose and α1,3-fucose epitopes 1277:1334 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 26868756 7 73 theme 85.5 1217:1220 arg1 % 1209:1209 % 1209:1209 % 1209:1209 The N-glycan structures that were presented on all of the four occupied N-glycosylation sites of recombinant GC in NbGNTI-RNAi plants (GC(gnt1) ) showed that the majority (ranging from 73.3% up to 85.5%) of the N-glycans had mannose-type structures lacking potential immunogenic β1,2-xylose and α1,3-fucose epitopes. 25697358 8 0 theme anomeric 1147:1154 arg1 center 1156:1161 the anomeric center 1143:1161 the anomeric center of the transferred GlcNAc-moiety 1143:1194 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 7 1 theme typical 1066:1072 arg1 fold 1083:1086 a typical Rossmann fold 1064:1086 a typical Rossmann fold 1064:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 2 theme Rossmann 1074:1081 arg1 fold 1083:1086 a typical Rossmann fold 1064:1086 a typical Rossmann fold 1064:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 3 contain contains 1011:1018 arg1 homotrimer 980:989 a propeller-like homotrimer 963:989 a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold 963:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 3 contain contains 1011:1018 arg2 domain 1052:1057 a GT-B-type glycosyltransferase domain 1020:1057 a GT-B-type glycosyltransferase domain with a typical Rossmann fold 1020:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 3 contain contains 1011:1018 arg1 blade 1005:1009 each blade 1000:1009 each blade 1000:1009 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 3 contain contains 1011:1018 arg2 blade 1005:1009 each blade 1000:1009 each blade 1000:1009 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 1 4 theme Anionic 173:179 arg1 glycopolymers 181:193 Anionic glycopolymers 173:193 Anionic glycopolymers known as wall teichoic acids (WTAs) 173:229 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 8 5 theme polyribitol 1203:1213 arg1 backbone 1215:1222 the polyribitol backbone 1199:1222 the polyribitol backbone 1199:1222 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 6 6 theme Å 922:922 arg1 resolution 924:933 2.8 Å resolution 918:933 2.8 Å resolution 918:933 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 5 7 theme α-O-N-acetylglycosylated 684:707 arg1 poly-5'-phosphoribitol 709:730 α-O-N-acetylglycosylated poly-5'-phosphoribitol 684:730 α-O-N-acetylglycosylated poly-5'-phosphoribitol 684:730 TarM performs the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure. 25697358 2 8 theme central 312:318 arg1 roles 320:324 central roles 312:324 central roles 312:324 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 1 9 theme many 273:276 arg1 bacteria 292:299 many Gram-positive bacteria 273:299 many Gram-positive bacteria 273:299 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 0 10 theme oligomeric 100:109 arg1 protein 111:117 an oligomeric protein 97:117 an oligomeric protein specific for the glycosylation of wall teichoic acid 97:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 5 11 theme poly-5'-phosphoribitol 709:730 arg1 synthesis 671:679 the synthesis 667:679 the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure 667:751 TarM performs the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure. 25697358 7 12 theme propeller-like 965:978 arg1 homotrimer 980:989 a propeller-like homotrimer 963:989 a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold 963:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 10 13 theme TarM 1362:1365 arg1 experiments 1347:1357 Structure-guided mutagenesis experiments 1318:1357 Structure-guided mutagenesis experiments of TarM 1318:1365 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 1 14 theme Gram-positive 278:290 arg1 bacteria 292:299 many Gram-positive bacteria 273:299 many Gram-positive bacteria 273:299 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 10 15 theme reaction 1501:1508 arg1 mechanism 1510:1518 a likely reaction mechanism 1492:1518 a likely reaction mechanism 1492:1518 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 10 16 theme mutagenesis 1335:1345 arg1 experiments 1347:1357 Structure-guided mutagenesis experiments 1318:1357 Structure-guided mutagenesis experiments of TarM 1318:1365 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 6 17 with complex 877:883 arg1 UDP-GlcNAc 904:913 its substrate UDP-GlcNAc 890:913 its substrate UDP-GlcNAc 890:913 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 0 18 theme specific 119:126 arg1 protein 111:117 an oligomeric protein 97:117 an oligomeric protein specific for the glycosylation of wall teichoic acid 97:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 10 19 theme likely 1494:1499 arg1 mechanism 1510:1518 a likely reaction mechanism 1492:1518 a likely reaction mechanism 1492:1518 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 8 20 from stereochemistry 1124:1138 arg1 backbone 1215:1222 the polyribitol backbone 1199:1222 the polyribitol backbone 1199:1222 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 10 21 theme Structure-guided 1318:1333 arg1 experiments 1347:1357 Structure-guided mutagenesis experiments 1318:1357 Structure-guided mutagenesis experiments of TarM 1318:1365 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 10 22 theme critical 1385:1392 arg1 residues 1376:1383 residues 1376:1383 residues critical for enzyme activity 1376:1412 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 6 23 theme substrate 894:902 arg1 UDP-GlcNAc 904:913 its substrate UDP-GlcNAc 890:913 its substrate UDP-GlcNAc 890:913 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 2 24 theme many 329:332 arg1 aspects 346:352 many fundamental aspects 329:352 many fundamental aspects of bacterial physiology 329:376 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 2 25 theme important 392:400 arg1 determinants 402:413 important determinants 392:413 important determinants of pathogenesis and antibiotic resistance 392:455 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 2 25 theme important 392:400 arg1 they 383:386 they 383:386 they 383:386 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 0 26 theme Structural 0:9 arg1 analysis 25:32 Structural and enzymatic analysis 0:32 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus 0:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 1 27 theme bacteria 292:299 arg1 layers 263:268 the peptidoglycan layers 245:268 the peptidoglycan layers of many Gram-positive bacteria 245:299 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 8 28 theme enzymatic 1093:1101 arg1 reaction 1103:1110 The enzymatic reaction 1089:1110 The enzymatic reaction 1089:1110 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 10 29 theme TarM 1453:1456 arg1 function 1458:1465 TarM function 1453:1465 TarM function 1453:1465 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 1 30 theme wall 204:207 arg1 acids 218:222 wall teichoic acids 204:222 wall teichoic acids (WTAs) 204:229 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 1 30 theme wall 204:207 arg1 WTAs 225:228 WTAs 225:228 WTAs 225:228 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 4 31 theme glycosyltransferase 574:592 arg1 TarM 594:597 the glycosyltransferase TarM 570:597 the glycosyltransferase TarM 570:597 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 4 31 theme glycosyltransferase 574:592 arg1 component 602:610 a component 600:610 a component of the WTA de novo biosynthesis pathway 600:650 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 9 32 theme novel 1262:1266 arg1 domain 1282:1287 a novel trimerization domain 1260:1287 a novel trimerization domain 1260:1287 TarM assembles into a trimer using a novel trimerization domain, here termed the HUB domain. 25697358 2 33 theme resistance 446:455 arg1 determinants 402:413 important determinants 392:413 important determinants of pathogenesis and antibiotic resistance 392:455 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 2 33 theme resistance 446:455 arg1 they 383:386 they 383:386 they 383:386 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 0 34 theme enzymatic 15:23 arg1 analysis 25:32 Structural and enzymatic analysis 0:32 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus 0:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 9 35 theme trimerization 1268:1280 arg1 domain 1282:1287 a novel trimerization domain 1260:1287 a novel trimerization domain 1260:1287 TarM assembles into a trimer using a novel trimerization domain, here termed the HUB domain. 25697358 2 36 theme physiology 367:376 arg1 aspects 346:352 many fundamental aspects 329:352 many fundamental aspects of bacterial physiology 329:376 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 8 37 theme GlcNAc-moiety 1182:1194 arg1 center 1156:1161 the anomeric center 1143:1161 the anomeric center of the transferred GlcNAc-moiety 1143:1194 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 2 38 theme antibiotic 435:444 arg1 resistance 446:455 antibiotic resistance 435:455 antibiotic resistance 435:455 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 0 39 theme TarM 37:40 arg1 glycosyltransferase 42:60 TarM glycosyltransferase 37:60 TarM glycosyltransferase from Staphylococcus aureus 37:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 10 40 from role 1433:1436 arg1 function 1458:1465 TarM function 1453:1465 TarM function 1453:1465 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 0 41 theme wall 153:156 arg1 acid 167:170 wall teichoic acid 153:170 wall teichoic acid 153:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 5 42 gly α-O-N-acetylglycosylated 684:707 arg1 poly-5'-phosphoribitol 709:730 α-O-N-acetylglycosylated poly-5'-phosphoribitol 684:730 α-O-N-acetylglycosylated poly-5'-phosphoribitol 684:730 TarM performs the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure. 25697358 3 43 theme enzymes 470:476 arg1 number 460:465 A number 458:465 A number of enzymes that glycosylate WTA in Staphylococcus aureus 458:522 A number of enzymes that glycosylate WTA in Staphylococcus aureus have recently been identified. 25697358 0 44 from aureus 82:87 arg1 analysis 25:32 Structural and enzymatic analysis 0:32 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus 0:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 0 44 from aureus 82:87 arg1 glycosyltransferase 42:60 TarM glycosyltransferase 37:60 TarM glycosyltransferase from Staphylococcus aureus 37:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 3 45 gly glycosylate 483:493 arg1 WTA 495:497 WTA 495:497 WTA 495:497 A number of enzymes that glycosylate WTA in Staphylococcus aureus have recently been identified. 25697358 6 46 theme Å 806:806 arg1 resolution 808:817 2.4 Å resolution 802:817 2.4 Å resolution 802:817 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 2 47 theme pathogenesis 418:429 arg1 determinants 402:413 important determinants 392:413 important determinants of pathogenesis and antibiotic resistance 392:455 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 2 47 theme pathogenesis 418:429 arg1 they 383:386 they 383:386 they 383:386 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 0 48 theme acid 167:170 arg1 glycosylation 136:148 the glycosylation 132:148 the glycosylation of wall teichoic acid 132:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 10 49 theme enzyme 1398:1403 arg1 activity 1405:1412 enzyme activity 1398:1412 enzyme activity 1398:1412 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 2 50 theme bacterial 357:365 arg1 physiology 367:376 bacterial physiology 357:376 bacterial physiology 357:376 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 0 51 theme glycosyltransferase 42:60 arg1 analysis 25:32 Structural and enzymatic analysis 0:32 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus 0:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 5 52 from synthesis 671:679 arg1 structure 743:751 the WTA structure 735:751 the WTA structure 735:751 TarM performs the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure. 25697358 0 53 theme teichoic 158:165 arg1 acid 167:170 wall teichoic acid 153:170 wall teichoic acid 153:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 6 54 theme enzyme 867:872 arg1 structure 850:858 a structure 848:858 a structure of the enzyme 848:872 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 7 55 theme GT-B-type 1022:1030 arg1 domain 1052:1057 a GT-B-type glycosyltransferase domain 1020:1057 a GT-B-type glycosyltransferase domain with a typical Rossmann fold 1020:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 0 56 gly glycosylation 136:148 arg1 acid 167:170 wall teichoic acid 153:170 wall teichoic acid 153:170 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 5 57 theme WTA 739:741 arg1 structure 743:751 the WTA structure 735:751 the WTA structure 735:751 TarM performs the synthesis of α-O-N-acetylglycosylated poly-5'-phosphoribitol in the WTA structure. 25697358 6 58 theme TarM 794:797 arg1 structure 781:789 the crystal structure 769:789 the crystal structure of TarM 769:797 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 7 59 with domain 1052:1057 arg1 fold 1083:1086 a typical Rossmann fold 1064:1086 a typical Rossmann fold 1064:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 7 60 theme glycosyltransferase 1032:1050 arg1 domain 1052:1057 a GT-B-type glycosyltransferase domain 1020:1057 a GT-B-type glycosyltransferase domain with a typical Rossmann fold 1020:1086 The protein assembles into a propeller-like homotrimer in which each blade contains a GT-B-type glycosyltransferase domain with a typical Rossmann fold. 25697358 9 61 theme HUB 1306:1308 arg1 domain 1310:1315 the HUB domain 1302:1315 the HUB domain 1302:1315 TarM assembles into a trimer using a novel trimerization domain, here termed the HUB domain. 25697358 10 62 theme putative 1424:1431 arg1 role 1433:1436 a putative role 1422:1436 a putative role for the HUB in TarM function 1422:1465 Structure-guided mutagenesis experiments of TarM identify residues critical for enzyme activity, assign a putative role for the HUB in TarM function, and allow us to propose a likely reaction mechanism. 25697358 2 63 theme fundamental 334:344 arg1 aspects 346:352 many fundamental aspects 329:352 many fundamental aspects of bacterial physiology 329:376 WTAs play central roles in many fundamental aspects of bacterial physiology, and they are important determinants of pathogenesis and antibiotic resistance. 25697358 4 64 theme de 623:624 arg1 pathway 644:650 the WTA de novo biosynthesis pathway 615:650 the WTA de novo biosynthesis pathway 615:650 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 8 65 theme transferred 1170:1180 arg1 GlcNAc-moiety 1182:1194 the transferred GlcNAc-moiety 1166:1194 the transferred GlcNAc-moiety 1166:1194 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 4 66 dep de 623:624 arg1 novo 626:629 novo 626:629 novo 626:629 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 1 67 theme teichoic 209:216 arg1 acids 218:222 wall teichoic acids 204:222 wall teichoic acids (WTAs) 204:229 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 1 67 theme teichoic 209:216 arg1 WTAs 225:228 WTAs 225:228 WTAs 225:228 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 25697358 4 68 theme pathway 644:650 arg1 TarM 594:597 the glycosyltransferase TarM 570:597 the glycosyltransferase TarM 570:597 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 4 68 theme pathway 644:650 arg1 component 602:610 a component 600:610 a component of the WTA de novo biosynthesis pathway 600:650 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 4 69 theme WTA 619:621 arg1 pathway 644:650 the WTA de novo biosynthesis pathway 615:650 the WTA de novo biosynthesis pathway 615:650 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 0 70 from analysis 25:32 arg1 aureus 82:87 Staphylococcus aureus 67:87 Staphylococcus aureus 67:87 Structural and enzymatic analysis of TarM glycosyltransferase from Staphylococcus aureus reveals an oligomeric protein specific for the glycosylation of wall teichoic acid. 25697358 4 71 theme biosynthesis 631:642 arg1 pathway 644:650 the WTA de novo biosynthesis pathway 615:650 the WTA de novo biosynthesis pathway 615:650 Among these is the glycosyltransferase TarM, a component of the WTA de novo biosynthesis pathway. 25697358 6 72 theme crystal 773:779 arg1 structure 781:789 the crystal structure 769:789 the crystal structure of TarM 769:797 We have solved the crystal structure of TarM at 2.4 Å resolution, and we have also determined a structure of the enzyme in complex with its substrate UDP-GlcNAc at 2.8 Å resolution. 25697358 8 73 theme center 1156:1161 arg1 stereochemistry 1124:1138 the stereochemistry 1120:1138 the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone 1120:1222 The enzymatic reaction retains the stereochemistry of the anomeric center of the transferred GlcNAc-moiety on the polyribitol backbone. 25697358 1 74 theme peptidoglycan 249:261 arg1 layers 263:268 the peptidoglycan layers 245:268 the peptidoglycan layers of many Gram-positive bacteria 245:299 Anionic glycopolymers known as wall teichoic acids (WTAs) functionalize the peptidoglycan layers of many Gram-positive bacteria. 28956227 6 0 with GlcNAc-Asn 1245:1254 arg1 treatment 1227:1235 kifunesine treatment 1216:1235 kifunesine treatment 1216:1235 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 10 1 theme evidence 2101:2108 arg1 mechanisms 2052:2061 the enzymatic mechanisms 2038:2061 the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities 2038:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 6 2 with treatment 1259:1267 arg1 EndoH 1274:1278 EndoH 1274:1278 EndoH 1274:1278 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 9 3 theme putative 1754:1761 arg1 structures 1763:1772 putative structures 1754:1772 putative structures derived from bacterial-originated CD39 domain proteins 1754:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 3 4 theme glycosylation 629:641 arg1 sites 643:647 seven N-linked glycosylation sites 614:647 seven N-linked glycosylation sites 614:647 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 5 5 theme ecto-NTPDase 957:968 arg1 activity 970:977 ecto-NTPDase activity 957:977 ecto-NTPDase activity 957:977 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 8 6 from Vmax 1592:1595 arg1 tri-phosphate 1618:1630 tri-phosphate 1618:1630 tri-phosphate 1618:1630 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 8 6 from Vmax 1592:1595 arg1 di- 1611:1613 di- 1611:1613 di- 1611:1613 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 1 7 theme glycosylated 230:241 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 5 8 from cells 1130:1134 arg1 domains 1077:1083 soluble enzymatic domains 1059:1083 soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1059:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 5 8 from cells 1130:1134 arg1 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1088:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 7 9 theme specific 1543:1550 arg1 activity 1560:1567 specific NTPDase activity 1543:1567 specific NTPDase activity 1543:1567 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 5 10 with NTPDase3/CD39L3 1088:1102 arg1 modifications 1163:1175 four different glycan modifications 1141:1175 four different glycan modifications 1141:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 1 11 theme extracellular 271:283 arg1 nucleosides 285:295 extracellular nucleosides di- and tri-phosphates 271:318 extracellular nucleosides di- and tri-phosphates 271:318 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 11 theme extracellular 271:283 arg1 tri-phosphates 305:318 tri-phosphates 305:318 tri-phosphates 305:318 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 11 theme extracellular 271:283 arg1 di- 297:299 di- 297:299 di- 297:299 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 9 12 attach derived 1774:1780 arg2 structures 1763:1772 putative structures 1754:1772 putative structures derived from bacterial-originated CD39 domain proteins 1754:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 9 12 attach derived 1774:1780 arg1 proteins 1820:1827 bacterial-originated CD39 domain proteins 1787:1827 bacterial-originated CD39 domain proteins 1787:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 5 13 theme glycan 1156:1161 arg1 modifications 1163:1175 four different glycan modifications 1141:1175 four different glycan modifications 1141:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 3 14 theme conserved 680:688 arg1 domains 690:696 four close to five highly conserved domains 654:696 four close to five highly conserved domains called "apyrase conserved regions" (ACRs) 654:738 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 15 theme regions 724:730 arg1 ACRs 734:737 ACRs 734:737 ACRs 734:737 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 15 theme regions 724:730 arg1 " 731:731 "apyrase conserved regions" 705:731 "apyrase conserved regions" (ACRs) 705:738 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 8 16 from Km 1605:1606 arg1 tri-phosphate 1618:1630 tri-phosphate 1618:1630 tri-phosphate 1618:1630 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 8 16 from Km 1605:1606 arg1 di- 1611:1613 di- 1611:1613 di- 1611:1613 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 3 17 theme apyrase 706:712 arg1 regions 724:730 apyrase conserved regions 706:730 "apyrase conserved regions" (ACRs) 705:738 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 5 18 theme specificity 993:1003 arg1 mechanism 940:948 the mechanism 936:948 the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans 936:1038 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 3 19 gly glycosylation 629:641 arg2 seven 614:618 seven 614:618 seven 614:618 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 19 gly glycosylation 629:641 arg2 sites 643:647 seven N-linked glycosylation sites 614:647 seven N-linked glycosylation sites 614:647 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 10 20 theme ectonucleotidase 2154:2169 arg1 activities 2171:2180 functional ectonucleotidase activities 2143:2180 functional ectonucleotidase activities 2143:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 0 21 theme domain 104:109 arg1 activity 57:64 E-NTPDase activity 47:64 E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain 47:109 Various N-glycoforms differentially upregulate E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain. 28956227 3 22 dep five 668:671 arg1 to 665:666 to 665:666 to 665:666 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 23 link N-linked 620:627 arg1 sites 643:647 seven N-linked glycosylation sites 614:647 seven N-linked glycosylation sites 614:647 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 24 contain has 610:612 arg2 sites 643:647 seven N-linked glycosylation sites 614:647 seven N-linked glycosylation sites 614:647 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 3 24 contain has 610:612 arg1 it 571:572 it 571:572 it 571:572 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 1 25 theme triphosphate 142:153 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 25 theme triphosphate 142:153 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 25 theme triphosphate 142:153 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 2 26 theme signaling 478:486 arg1 responses 488:496 purinergic signaling responses 467:496 purinergic signaling responses 467:496 This catalytic process differentially regulates a complex array of purinergic signaling responses. 28956227 5 27 theme NTPDase3/CD39L3 1088:1102 arg1 domains 1077:1083 soluble enzymatic domains 1059:1083 soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1059:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 5 27 theme NTPDase3/CD39L3 1088:1102 arg1 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1088:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 0 28 theme ecto-enzymatic 89:102 arg1 domain 104:109 the NTPDase3/CD39L3 ecto-enzymatic domain 69:109 the NTPDase3/CD39L3 ecto-enzymatic domain 69:109 Various N-glycoforms differentially upregulate E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain. 28956227 4 29 theme preferential 802:813 arg1 substrate 815:823 its preferential substrate 798:823 its preferential substrate 798:823 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 0 30 theme Various 0:6 arg1 N-glycoforms 8:19 Various N-glycoforms 0:19 Various N-glycoforms 0:19 Various N-glycoforms differentially upregulate E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain. 28956227 1 31 theme divalent 339:346 arg1 cations 348:354 divalent cations 339:354 divalent cations 339:354 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 32 theme GDA1/CD39 116:124 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 32 theme GDA1/CD39 116:124 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 32 theme GDA1/CD39 116:124 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 5 33 theme soluble 1059:1065 arg1 domains 1077:1083 soluble enzymatic domains 1059:1083 soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1059:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 5 33 theme soluble 1059:1065 arg1 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1088:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 5 34 dep activity 970:977 arg1 the 953:955 the 953:955 the 953:955 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 0 35 theme E-NTPDase 47:55 arg1 activity 57:64 E-NTPDase activity 47:64 E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain 47:109 Various N-glycoforms differentially upregulate E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain. 28956227 0 36 theme NTPDase3/CD39L3 73:87 arg1 domain 104:109 the NTPDase3/CD39L3 ecto-enzymatic domain 69:109 the NTPDase3/CD39L3 ecto-enzymatic domain 69:109 Various N-glycoforms differentially upregulate E-NTPDase activity of the NTPDase3/CD39L3 ecto-enzymatic domain. 28956227 3 37 theme insulin 587:593 arg1 secretion 595:603 insulin secretion 587:603 insulin secretion 587:603 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 1 38 theme monophosphate 373:385 arg1 derivatives 387:397 the monophosphate derivatives 369:397 the monophosphate derivatives 369:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 5 39 theme different 1146:1154 arg1 modifications 1163:1175 four different glycan modifications 1141:1175 four different glycan modifications 1141:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 10 40 theme members 2081:2087 arg1 mechanisms 2052:2061 the enzymatic mechanisms 2038:2061 the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities 2038:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 4 41 theme other 874:878 arg1 triphosphate 880:891 other triphosphate and diphosphate nucleosides 874:919 triphosphate 880:891 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 5 42 theme human 1107:1111 arg1 cells 1130:1134 human embryotic kidney cells 1107:1134 human embryotic kidney cells 1107:1134 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 7 43 theme ecto-enzymatic 1423:1436 arg1 domain 1438:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 10 44 theme NTPDase 2066:2072 arg1 members 2081:2087 NTPDase family members 2066:2087 NTPDase family members 2066:2087 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 2 45 theme complex 450:456 arg1 array 458:462 a complex array 448:462 a complex array of purinergic signaling responses 448:496 This catalytic process differentially regulates a complex array of purinergic signaling responses. 28956227 5 46 theme kidney 1123:1128 arg1 cells 1130:1134 human embryotic kidney cells 1107:1134 human embryotic kidney cells 1107:1134 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 7 47 theme non-glycosylated 1390:1405 arg1 domain 1438:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 5 48 from NTPDase3/CD39L3 1088:1102 arg1 cells 1130:1134 human embryotic kidney cells 1107:1134 human embryotic kidney cells 1107:1134 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 9 49 theme N-glycan 1843:1850 arg1 modifications 1852:1864 N-glycan modifications 1843:1864 N-glycan modifications 1843:1864 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 6 50 theme kifunesine 1216:1225 arg1 treatment 1227:1235 kifunesine treatment 1216:1235 kifunesine treatment 1216:1235 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 10 51 theme further 2093:2099 arg1 evidence 2101:2108 further evidence 2093:2108 further evidence that N-glycans directly modulate functional ectonucleotidase activities 2093:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 1 52 gly glycosylated 230:241 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 5 53 from domains 1077:1083 arg1 cells 1130:1134 human embryotic kidney cells 1107:1134 human embryotic kidney cells 1107:1134 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 10 54 theme functional 2143:2152 arg1 activities 2171:2180 functional ectonucleotidase activities 2143:2180 functional ectonucleotidase activities 2143:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 3 55 theme N-linked 620:627 arg1 sites 643:647 seven N-linked glycosylation sites 614:647 seven N-linked glycosylation sites 614:647 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 6 56 gly de-glycosylated 1281:1295 arg1 form 1297:1300 de-glycosylated form 1281:1300 de-glycosylated form by treatment with PNGaseF 1281:1326 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 5 57 theme activity 970:977 arg1 mechanism 940:948 the mechanism 936:948 the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans 936:1038 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 6 58 with treatment 1305:1313 arg1 PNGaseF 1320:1326 PNGaseF 1320:1326 PNGaseF 1320:1326 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 6 59 theme de-glycosylated 1281:1295 arg1 form 1297:1300 de-glycosylated form 1281:1300 de-glycosylated form by treatment with PNGaseF 1281:1326 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 9 60 theme bacterial-originated 1787:1806 arg1 proteins 1820:1827 bacterial-originated CD39 domain proteins 1787:1827 bacterial-originated CD39 domain proteins 1787:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 1 61 theme ecto-enzymes 243:254 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 61 theme ecto-enzymes 243:254 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 61 theme ecto-enzymes 243:254 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 9 62 from Asn149 1869:1874 arg1 modifications 1852:1864 N-glycan modifications 1843:1864 N-glycan modifications 1843:1864 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 9 63 theme N-terminal 1904:1913 arg1 modifications 1852:1864 N-glycan modifications 1843:1864 N-glycan modifications 1843:1864 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 1 64 dep nucleosides 285:295 arg1 nucleosides 285:295 extracellular nucleosides di- and tri-phosphates 271:318 extracellular nucleosides di- and tri-phosphates 271:318 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 64 dep nucleosides 285:295 arg1 tri-phosphates 305:318 tri-phosphates 305:318 tri-phosphates 305:318 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 64 dep nucleosides 285:295 arg1 di- 297:299 di- 297:299 di- 297:299 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 7 65 theme NTPDase 1552:1558 arg1 activity 1560:1567 specific NTPDase activity 1543:1567 specific NTPDase activity 1543:1567 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 9 66 theme catalytic 1961:1969 arg1 pocket 1971:1976 the catalytic pocket 1957:1976 the catalytic pocket of NTPDase3/CD39L3 1957:1995 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 1 67 theme diphosphosphohydrolase 155:176 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 67 theme diphosphosphohydrolase 155:176 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 67 theme diphosphosphohydrolase 155:176 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 6 68 with form 1297:1300 arg1 treatment 1227:1235 kifunesine treatment 1216:1235 kifunesine treatment 1216:1235 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 4 69 contain possesses 834:842 arg2 activities 856:865 significant activities 844:865 significant activities toward other triphosphate and diphosphate nucleosides 844:919 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 4 69 contain possesses 834:842 arg1 NTPDase3/CD39L3 770:784 NTPDase3/CD39L3 770:784 NTPDase3/CD39L3 770:784 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 3 70 theme conserved 714:722 arg1 regions 724:730 apyrase conserved regions 706:730 "apyrase conserved regions" (ACRs) 705:738 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 1 71 theme E-NTPDase 179:187 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 71 theme E-NTPDase 179:187 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 71 theme E-NTPDase 179:187 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 9 72 theme critical 1928:1935 arg1 roles 1937:1941 critical roles 1928:1941 critical roles 1928:1941 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 6 73 theme wild-type 1333:1341 arg1 glycans 1343:1349 wild-type glycans 1333:1349 wild-type glycans 1333:1349 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 7 74 theme functional 1356:1365 arg1 data 1367:1370 Our functional data 1352:1370 Our functional data 1352:1370 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 10 75 theme new 2020:2022 arg1 insights 2024:2031 both new insights 2015:2031 both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities 2015:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 5 76 theme substrate 983:991 arg1 specificity 993:1003 substrate specificity 983:1003 substrate specificity 983:1003 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 4 77 theme similar 753:759 arg1 manner 746:751 a manner 744:751 a manner similar to CD39 744:767 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 9 78 theme domain 1813:1818 arg1 proteins 1820:1827 bacterial-originated CD39 domain proteins 1787:1827 bacterial-originated CD39 domain proteins 1787:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 5 79 theme enzymatic 1067:1075 arg1 domains 1077:1083 soluble enzymatic domains 1059:1083 soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1059:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 5 79 theme enzymatic 1067:1075 arg1 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 1088:1102 NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications 1088:1175 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 6 80 with glycans 1343:1349 arg1 treatment 1227:1235 kifunesine treatment 1216:1235 kifunesine treatment 1216:1235 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 9 81 theme Structural 1716:1725 arg1 analysis 1736:1743 Structural modeling analysis 1716:1743 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins 1716:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 9 82 theme NTPDase3/CD39L3 1981:1995 arg1 pocket 1971:1976 the catalytic pocket 1957:1976 the catalytic pocket of NTPDase3/CD39L3 1957:1995 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 8 83 theme glycan 1696:1701 arg1 attachments 1703:1713 the glycan attachments 1692:1713 the glycan attachments 1692:1713 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 4 84 theme significant 844:854 arg1 activities 856:865 significant activities 844:865 significant activities toward other triphosphate and diphosphate nucleosides 844:919 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 2 85 theme purinergic 467:476 arg1 responses 488:496 purinergic signaling responses 467:496 purinergic signaling responses 467:496 This catalytic process differentially regulates a complex array of purinergic signaling responses. 28956227 7 86 theme N-glycan 1472:1479 arg1 GlcNAc-Asn 1506:1515 the GlcNAc-Asn 1502:1515 the GlcNAc-Asn 1502:1515 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 7 86 theme N-glycan 1472:1479 arg1 attachments 1481:1491 N-glycan attachments 1472:1491 N-glycan attachments 1472:1491 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 1 87 theme cations 348:354 arg1 presence 327:334 the presence 323:334 the presence of divalent cations 323:354 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 7 88 gly non-glycosylated 1390:1405 arg1 domain 1438:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 1 89 theme ecto-nucleoside 126:140 arg1 ecto-enzymes 243:254 eight heavily glycosylated ecto-enzymes 216:254 eight heavily glycosylated ecto-enzymes 216:254 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 89 theme ecto-nucleoside 126:140 arg1 superfamily 190:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily 112:200 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 1 89 theme ecto-nucleoside 126:140 arg1 group 207:211 a group 205:211 a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives 205:397 The GDA1/CD39 ecto-nucleoside triphosphate diphosphosphohydrolase (E-NTPDase) superfamily is a group of eight heavily glycosylated ecto-enzymes that hydrolyze extracellular nucleosides di- and tri-phosphates in the presence of divalent cations, to generate the monophosphate derivatives. 28956227 8 90 dep di- 1611:1613 arg1 nucleosides 1632:1642 nucleosides 1632:1642 nucleosides 1632:1642 Both the Vmax and the Km on di- or tri-phosphate nucleosides are substantially and differentially altered by the glycan attachments. 28956227 3 91 theme pancreatic 541:550 arg1 cells 558:562 pancreatic islet cells 541:562 pancreatic islet cells 541:562 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 10 92 theme enzymatic 2042:2050 arg1 mechanisms 2052:2061 the enzymatic mechanisms 2038:2061 the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities 2038:2180 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 4 93 theme diphosphate 897:907 arg1 nucleosides 909:919 other triphosphate and diphosphate nucleosides 874:919 nucleosides 909:919 In a manner similar to CD39, NTPDase3/CD39L3 uses ATP as its preferential substrate and also possesses significant activities toward other triphosphate and diphosphate nucleosides. 28956227 2 94 theme responses 488:496 arg1 array 458:462 a complex array 448:462 a complex array of purinergic signaling responses 448:496 This catalytic process differentially regulates a complex array of purinergic signaling responses. 28956227 6 95 with glycans 1203:1209 arg1 treatment 1227:1235 kifunesine treatment 1216:1235 kifunesine treatment 1216:1235 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 2 96 theme catalytic 405:413 arg1 process 415:421 This catalytic process 400:421 This catalytic process 400:421 This catalytic process differentially regulates a complex array of purinergic signaling responses. 28956227 9 97 theme CD39 1808:1811 arg1 proteins 1820:1827 bacterial-originated CD39 domain proteins 1787:1827 bacterial-originated CD39 domain proteins 1787:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 7 98 theme NTPDase3/CD39L3 1407:1421 arg1 domain 1438:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain 1386:1443 Our functional data indicate that the non-glycosylated NTPDase3/CD39L3 ecto-enzymatic domain retains activity, but that N-glycan attachments, such as the GlcNAc-Asn, substantially upregulate specific NTPDase activity by 2-20 fold. 28956227 9 99 theme modeling 1727:1734 arg1 analysis 1736:1743 Structural modeling analysis 1716:1743 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins 1716:1827 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 10 100 theme family 2074:2079 arg1 members 2081:2087 NTPDase family members 2066:2087 NTPDase family members 2066:2087 Our data provide both new insights into the enzymatic mechanisms of NTPDase family members and further evidence that N-glycans directly modulate functional ectonucleotidase activities. 28956227 5 101 theme embryotic 1113:1121 arg1 cells 1130:1134 human embryotic kidney cells 1107:1134 human embryotic kidney cells 1107:1134 To understand the mechanism of the ecto-NTPDase activity and substrate specificity, potentially impacted by N-glycans, we have generated soluble enzymatic domains of NTPDase3/CD39L3 in human embryotic kidney cells with four different glycan modifications. 28956227 6 102 theme single 1238:1243 arg1 GlcNAc-Asn 1245:1254 single GlcNAc-Asn 1238:1254 single GlcNAc-Asn by treatment with EndoH 1238:1278 These include mannose5-9 glycans with kifunesine treatment, single GlcNAc-Asn by treatment with EndoH, de-glycosylated form by treatment with PNGaseF, and wild-type glycans. 28956227 3 103 theme islet 552:556 arg1 cells 558:562 pancreatic islet cells 541:562 pancreatic islet cells 541:562 NTPDase3/CD39L3is dominantly expressed in pancreatic islet cells, where it may regulate insulin secretion, and has seven N-linked glycosylation sites with four close to five highly conserved domains called "apyrase conserved regions" (ACRs). 28956227 9 104 contain have 1923:1926 arg1 modifications 1852:1864 N-glycan modifications 1843:1864 N-glycan modifications 1843:1864 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28956227 9 104 contain have 1923:1926 arg2 roles 1937:1941 critical roles 1928:1941 critical roles 1928:1941 Structural modeling analysis based on putative structures derived from bacterial-originated CD39 domain proteins suggests that N-glycan modifications at Asn149 next to ACR2 and/or Asn454, N-terminal to ACR5 have critical roles in regulating the catalytic pocket of NTPDase3/CD39L3. 28289219 7 0 theme core 1228:1231 arg1 fucosylation 1233:1244 core fucosylation 1228:1244 core fucosylation 1228:1244 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 7 1 from effect 1167:1172 arg1 ADCC 1192:1195 ADCC 1192:1195 ADCC 1192:1195 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 5 2 dep in 857:858 arg1 vitro 860:864 vitro 860:864 vitro 860:864 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 6 3 theme in 1075:1076 arg1 depletion 1105:1113 in vivo IgG-mediated cellular depletion 1075:1113 in vivo IgG-mediated cellular depletion 1075:1113 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 5 4 theme IgG-mediated 925:936 arg1 studies 957:963 in vivo IgG-mediated cellular depletion studies 917:963 in vivo IgG-mediated cellular depletion studies 917:963 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 7 5 from dependent 1201:1209 arg1 contrast 1153:1160 contrast 1153:1160 contrast 1153:1160 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 4 6 theme specific 585:592 arg1 combinations 594:605 specific combinations 585:605 specific combinations of sugars in the glycan 585:629 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 4 7 from sugars 610:615 arg1 glycan 624:629 the glycan 620:629 the glycan 620:629 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 5 8 theme fucosylation 801:812 arg1 combinations 785:796 different combinations 775:796 different combinations of fucosylation and sialylation 775:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 1 9 theme complex 143:149 arg1 glycan 164:169 a complex, biantennary glycan 141:169 a complex, biantennary glycan 141:169 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 6 10 theme FcγRIIIA 1038:1045 arg1 binding 1047:1053 FcγRIIIA binding 1038:1053 FcγRIIIA binding 1038:1053 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 5 11 theme homogeneous 733:743 arg1 glycoforms 745:754 structurally well-defined homogeneous glycoforms 707:754 structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation 707:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 2 12 theme binding 273:279 arg1 affinity 281:288 the binding affinity 269:288 the binding affinity of the Fc to diverse Fc receptor family members 269:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 9 13 theme fucosylation 1432:1443 arg1 absence 1421:1427 the absence 1417:1427 the absence of fucosylation 1417:1443 In contrast, in the absence of fucosylation, sialylation did not adversely impact ADCC. 28289219 4 14 from combinations 594:605 arg1 glycan 624:629 the glycan 620:629 the glycan 620:629 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 6 15 dep in 1056:1057 arg1 vitro 1059:1063 vitro 1059:1063 vitro 1059:1063 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 3 16 theme immunoglobulin 547:560 arg1 therapy 562:568 human i.v. immunoglobulin therapy 536:568 human i.v. immunoglobulin therapy 536:568 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 1 17 theme IgG 59:61 arg1 antibodies 63:72 IgG antibodies 59:72 IgG antibodies 59:72 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 4 18 theme combinations 594:605 arg1 effect 575:580 The effect 571:580 The effect of specific combinations of sugars in the glycan on ADCC 571:637 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 3 19 theme critical 485:492 arg1 role 494:497 a critical role 483:497 a critical role 483:497 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 4 20 from effect 575:580 arg1 ADCC 634:637 ADCC 634:637 ADCC 634:637 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 5 21 dep in 917:918 arg1 vivo 920:923 vivo 920:923 vivo 920:923 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 9 22 theme impact 1476:1481 arg1 ADCC 1483:1486 impact ADCC 1476:1486 impact ADCC 1476:1486 In contrast, in the absence of fucosylation, sialylation did not adversely impact ADCC. 28289219 5 23 theme ADCC 900:903 arg1 assays 905:910 cell-based ADCC assays 889:910 cell-based ADCC assays 889:910 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 1 24 gly N-glycosylation 94:108 arg2 site 110:113 a conserved N-glycosylation site 82:113 a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached 82:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 5 25 theme cellular 938:945 arg1 studies 957:963 in vivo IgG-mediated cellular depletion studies 917:963 in vivo IgG-mediated cellular depletion studies 917:963 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 3 26 theme antibody-dependent 394:411 arg1 ADCC 436:439 ADCC 436:439 ADCC 436:439 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 3 26 theme antibody-dependent 394:411 arg1 cytotoxicity 422:433 antibody-dependent cellular cytotoxicity 394:433 antibody-dependent cellular cytotoxicity (ADCC) 394:440 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 3 27 theme human 536:540 arg1 therapy 562:568 human i.v. immunoglobulin therapy 536:568 human i.v. immunoglobulin therapy 536:568 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 7 28 from contrast 1153:1160 arg1 effect 1167:1172 the effect 1163:1172 the effect of sialylation on ADCC 1163:1195 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 7 28 from contrast 1153:1160 arg1 dependent 1201:1209 dependent 1201:1209 dependent 1201:1209 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 0 29 theme IgG 11:13 arg1 function 24:31 IgG effector function 11:31 IgG effector function 11:31 Modulating IgG effector function by Fc glycan engineering. 28289219 7 30 theme fucosylation 1233:1244 arg1 status 1218:1223 the status 1214:1223 the status of core fucosylation 1214:1244 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 6 31 theme adverse 1020:1026 arg1 effect 1028:1033 a significant adverse effect 1006:1033 a significant adverse effect 1006:1033 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 2 32 theme Fc 297:298 arg1 affinity 281:288 the binding affinity 269:288 the binding affinity of the Fc to diverse Fc receptor family members 269:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 5 33 gly glycoforms 745:754 arg1 antibodies 759:768 antibodies 759:768 antibodies with different combinations of fucosylation and sialylation 759:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 6 34 theme cellular 1096:1103 arg1 depletion 1105:1113 in vivo IgG-mediated cellular depletion 1075:1113 in vivo IgG-mediated cellular depletion 1075:1113 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 5 35 theme side-by-side 844:855 arg1 analyses 879:886 side-by-side in vitro FcγR-binding analyses 844:886 side-by-side in vitro FcγR-binding analyses 844:886 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 1 36 contain contain 74:80 arg1 antibodies 63:72 IgG antibodies 59:72 IgG antibodies 59:72 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 1 36 contain contain 74:80 arg2 site 110:113 a conserved N-glycosylation site 82:113 a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached 82:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 1 37 theme conserved 84:92 arg1 site 110:113 a conserved N-glycosylation site 82:113 a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached 82:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 8 38 theme core 1277:1280 arg1 fucosylation 1282:1293 core fucosylation 1277:1293 core fucosylation 1277:1293 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 6 39 theme IgG-mediated 1083:1094 arg1 depletion 1105:1113 in vivo IgG-mediated cellular depletion 1075:1113 in vivo IgG-mediated cellular depletion 1075:1113 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 4 40 theme sugars 610:615 arg1 combinations 594:605 specific combinations 585:605 specific combinations of sugars in the glycan 585:629 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 6 41 theme significant 1008:1018 arg1 effect 1028:1033 a significant adverse effect 1006:1033 a significant adverse effect 1006:1033 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 1 42 theme N-glycosylation 94:108 arg1 site 110:113 a conserved N-glycosylation site 82:113 a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached 82:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 3 43 theme terminal 451:458 arg1 α2,6-sialylation 460:475 terminal α2,6-sialylation 451:475 terminal α2,6-sialylation 451:475 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 1 44 dep complex 143:149 arg1 biantennary 152:162 biantennary 152:162 biantennary 152:162 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 8 45 theme fucosylation 1282:1293 arg1 context 1266:1272 the context 1262:1272 the context of core fucosylation 1262:1293 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 2 46 theme family 323:328 arg1 members 330:336 diverse Fc receptor family members 303:336 diverse Fc receptor family members 303:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 7 47 theme sialylation 1177:1187 arg1 effect 1167:1172 the effect 1163:1172 the effect of sialylation on ADCC 1163:1195 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 7 47 theme sialylation 1177:1187 arg1 dependent 1201:1209 dependent 1201:1209 dependent 1201:1209 In contrast, the effect of sialylation on ADCC was dependent on the status of core fucosylation. 28289219 5 48 theme in 917:918 arg1 studies 957:963 in vivo IgG-mediated cellular depletion studies 917:963 in vivo IgG-mediated cellular depletion studies 917:963 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 0 49 theme Fc 36:37 arg1 engineering 46:56 Fc glycan engineering 36:56 Fc glycan engineering 36:56 Modulating IgG effector function by Fc glycan engineering. 28289219 2 50 theme glycan 212:217 arg1 structures 193:202 The fine structures 184:202 The fine structures of this glycan 184:217 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 8 51 theme cell-based 1329:1338 arg1 assay 1340:1344 a cell-based assay 1327:1344 a cell-based assay 1327:1344 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 3 52 theme cellular 413:420 arg1 ADCC 436:439 ADCC 436:439 ADCC 436:439 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 3 52 theme cellular 413:420 arg1 cytotoxicity 422:433 antibody-dependent cellular cytotoxicity 394:433 antibody-dependent cellular cytotoxicity (ADCC) 394:440 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 5 53 theme well-defined 720:731 arg1 glycoforms 745:754 structurally well-defined homogeneous glycoforms 707:754 structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation 707:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 5 54 theme in 857:858 arg1 analyses 879:886 side-by-side in vitro FcγR-binding analyses 844:886 side-by-side in vitro FcγR-binding analyses 844:886 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 8 55 from Sialylation 1247:1257 arg1 context 1266:1272 the context 1262:1272 the context of core fucosylation 1262:1293 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 5 56 theme antibodies 759:768 arg1 glycoforms 745:754 structurally well-defined homogeneous glycoforms 707:754 structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation 707:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 5 57 theme sialylation 818:828 arg1 combinations 785:796 different combinations 775:796 different combinations of fucosylation and sialylation 775:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 6 58 theme core 980:983 arg1 fucosylation 985:996 core fucosylation 980:996 core fucosylation 980:996 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 2 59 theme receptor 314:321 arg1 members 330:336 diverse Fc receptor family members 303:336 diverse Fc receptor family members 303:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 0 60 theme glycan 39:44 arg1 engineering 46:56 Fc glycan engineering 36:56 Fc glycan engineering 36:56 Modulating IgG effector function by Fc glycan engineering. 28289219 3 61 theme therapy 562:568 arg1 activity 524:531 the anti-inflammatory activity 502:531 the anti-inflammatory activity of human i.v. immunoglobulin therapy 502:568 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 2 62 theme fine 188:191 arg1 structures 193:202 The fine structures 184:202 The fine structures of this glycan 184:217 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 2 63 theme Fc 311:312 arg1 members 330:336 diverse Fc receptor family members 303:336 diverse Fc receptor family members 303:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 1 64 attach attached 174:181 arg1 domain 125:130 the Fc domain 118:130 the Fc domain to which a complex, biantennary glycan is attached 118:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 1 64 attach attached 174:181 arg2 glycan 164:169 a complex, biantennary glycan 141:169 a complex, biantennary glycan 141:169 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 5 65 with antibodies 759:768 arg1 combinations 785:796 different combinations 775:796 different combinations of fucosylation and sialylation 775:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 1 66 theme Fc 122:123 arg1 domain 125:130 the Fc domain 118:130 the Fc domain to which a complex, biantennary glycan is attached 118:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 1 67 from site 110:113 arg1 domain 125:130 the Fc domain 118:130 the Fc domain to which a complex, biantennary glycan is attached 118:181 IgG antibodies contain a conserved N-glycosylation site on the Fc domain to which a complex, biantennary glycan is attached. 28289219 6 68 dep in 1075:1076 arg1 vivo 1078:1081 vivo 1078:1081 vivo 1078:1081 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 5 69 theme depletion 947:955 arg1 studies 957:963 in vivo IgG-mediated cellular depletion studies 917:963 in vivo IgG-mediated cellular depletion studies 917:963 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 2 70 theme effector 237:244 arg1 functions 246:254 antibody effector functions 228:254 antibody effector functions 228:254 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 6 71 theme sialylation 1130:1140 arg1 status 1142:1147 sialylation status 1130:1147 sialylation status 1130:1147 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 8 72 theme cell 1379:1382 arg1 killing 1384:1390 antibody-mediated cell killing 1361:1390 antibody-mediated cell killing 1361:1390 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 2 73 theme diverse 303:309 arg1 members 330:336 diverse Fc receptor family members 303:336 diverse Fc receptor family members 303:336 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 4 74 from glycan 624:629 arg1 combinations 594:605 specific combinations 585:605 specific combinations of sugars in the glycan 585:629 The effect of specific combinations of sugars in the glycan on ADCC remains to be further addressed, however. 28289219 5 75 theme cell-based 889:898 arg1 assays 905:910 cell-based ADCC assays 889:910 cell-based ADCC assays 889:910 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 2 76 theme antibody 228:235 arg1 functions 246:254 antibody effector functions 228:254 antibody effector functions 228:254 The fine structures of this glycan modulate antibody effector functions by affecting the binding affinity of the Fc to diverse Fc receptor family members. 28289219 5 77 theme FcγR-binding 866:877 arg1 analyses 879:886 side-by-side in vitro FcγR-binding analyses 844:886 side-by-side in vitro FcγR-binding analyses 844:886 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 3 78 theme anti-inflammatory 506:522 arg1 activity 524:531 the anti-inflammatory activity 502:531 the anti-inflammatory activity of human i.v. immunoglobulin therapy 502:568 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 6 79 theme in 1056:1057 arg1 ADCC 1065:1068 in vitro ADCC 1056:1068 in vitro ADCC 1056:1068 We found that core fucosylation exerted a significant adverse effect on FcγRIIIA binding, in vitro ADCC, and in vivo IgG-mediated cellular depletion, regardless of sialylation status. 28289219 5 80 theme different 775:783 arg1 combinations 785:796 different combinations 775:796 different combinations of fucosylation and sialylation 775:828 Therefore, we synthesized structurally well-defined homogeneous glycoforms of antibodies with different combinations of fucosylation and sialylation and performed side-by-side in vitro FcγR-binding analyses, cell-based ADCC assays, and in vivo IgG-mediated cellular depletion studies. 28289219 3 81 theme i.v. 542:545 arg1 therapy 562:568 human i.v. immunoglobulin therapy 536:568 human i.v. immunoglobulin therapy 536:568 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 28289219 0 82 theme effector 15:22 arg1 function 24:31 IgG effector function 11:31 IgG effector function 11:31 Modulating IgG effector function by Fc glycan engineering. 28289219 8 83 theme antibody-mediated 1361:1377 arg1 killing 1384:1390 antibody-mediated cell killing 1361:1390 antibody-mediated cell killing 1361:1390 Sialylation in the context of core fucosylation significantly decreased ADCC in a cell-based assay and suppressed antibody-mediated cell killing in vivo. 28289219 3 84 theme core 352:355 arg1 fucosylation 357:368 core fucosylation 352:368 core fucosylation 352:368 For example, core fucosylation significantly decreases antibody-dependent cellular cytotoxicity (ADCC), whereas terminal α2,6-sialylation plays a critical role in the anti-inflammatory activity of human i.v. immunoglobulin therapy. 25643168 2 0 theme motifs 474:479 arg1 presence 449:456 the presence 445:456 the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion 445:611 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 4 1 dep high 894:897 arg1 > 902:902 R > 0.98 900:907 R > 0.98 900:907 Reproducibility was extraordinarily high (R > 0.98) for both method and instrumental replicates. 25643168 2 2 theme structures 343:352 arg1 Hundreds 324:331 Hundreds 324:331 Hundreds of glycan structures 324:352 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 5 3 theme quantitative 1080:1091 arg1 exploration 1093:1103 qualitative and quantitative exploration 1064:1103 qualitative and quantitative exploration of the brain glycome 1064:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 2 4 theme glycan 467:472 arg1 glucuronidation 528:542 glucuronidation 528:542 glucuronidation 528:542 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 4 theme glycan 467:472 arg1 fucosylation 499:510 antennary fucosylation 489:510 antennary fucosylation 489:510 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 4 theme glycan 467:472 arg1 motifs 474:479 novel glycan motifs 461:479 novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion 461:611 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 4 theme glycan 467:472 arg1 sulfation 513:521 sulfation 513:521 sulfation 513:521 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 5 5 theme qualitative 1064:1074 arg1 exploration 1093:1103 qualitative and quantitative exploration 1064:1103 qualitative and quantitative exploration of the brain glycome 1064:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 2 6 theme glycan 336:341 arg1 structures 343:352 glycan structures 336:352 glycan structures 336:352 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 1 7 theme structure-specific 229:246 arg1 nano-LC/MS 248:257 structure-specific nano-LC/MS 229:257 structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome 229:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 8 theme isolated 807:814 arg1 structures 822:831 isolated brain structures 807:831 not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus) 721:855 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 3 9 theme brain 730:734 arg1 sections 736:743 brain sections 730:743 not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus) 721:855 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 2 10 theme cellular 581:588 arg1 signaling 590:598 cellular signaling 581:598 cellular signaling 581:598 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 3 11 theme brain 816:820 arg1 structures 822:831 isolated brain structures 807:831 not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus) 721:855 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 5 12 with pairing 959:965 arg1 nano-LC/MS 998:1007 structure-specific nano-LC/MS 979:1007 structure-specific nano-LC/MS 979:1007 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 3 13 dep hippocampus 844:854 arg1 e.g. 834:837 e.g. 834:837 e.g. 834:837 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 1 14 theme sensitive 141:149 arg1 method 165:170 a highly sensitive MS-compatible method 132:170 a highly sensitive MS-compatible method for extraction of glycans from tissue 132:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 14 theme sensitive 141:149 arg1 glyco-capture 111:123 Tissue glyco-capture 104:123 Tissue glyco-capture (TGC) 104:129 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 2 15 theme antennary 489:497 arg1 fucosylation 499:510 antennary fucosylation 489:510 antennary fucosylation 489:510 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 16 theme novel 461:465 arg1 glucuronidation 528:542 glucuronidation 528:542 glucuronidation 528:542 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 16 theme novel 461:465 arg1 fucosylation 499:510 antennary fucosylation 489:510 antennary fucosylation 489:510 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 16 theme novel 461:465 arg1 motifs 474:479 novel glycan motifs 461:479 novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion 461:611 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 2 16 theme novel 461:465 arg1 sulfation 513:521 sulfation 513:521 sulfation 513:521 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 1 17 theme MS-compatible 151:163 arg1 method 165:170 a highly sensitive MS-compatible method 132:170 a highly sensitive MS-compatible method for extraction of glycans from tissue 132:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 17 theme MS-compatible 151:163 arg1 glyco-capture 111:123 Tissue glyco-capture 104:123 Tissue glyco-capture (TGC) 104:129 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 0 18 theme Spatially-resolved 0:17 arg1 exploration 19:29 Spatially-resolved exploration 0:29 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 0:102 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 1 19 theme sensitive 263:271 arg1 profiling 286:294 sensitive and detailed profiling 263:294 sensitive and detailed profiling of the mouse brain glycome 263:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 20 theme one-dimensional 753:767 arg1 resolution 777:786 a one-dimensional spatial resolution 751:786 a one-dimensional spatial resolution of 20 μm 751:795 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 4 21 dep instrumental 930:941 arg1 replicates 943:952 replicates 943:952 replicates 943:952 Reproducibility was extraordinarily high (R > 0.98) for both method and instrumental replicates. 25643168 1 22 theme detailed 277:284 arg1 profiling 286:294 sensitive and detailed profiling 263:294 sensitive and detailed profiling of the mouse brain glycome 263:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 23 with structures 822:831 arg1 resolution 777:786 a one-dimensional spatial resolution 751:786 a one-dimensional spatial resolution of 20 μm 751:795 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 0 24 theme brain 44:48 arg1 glycome 50:56 the mouse brain glycome 34:56 the mouse brain glycome 34:56 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 5 25 theme structure-specific 979:996 arg1 nano-LC/MS 998:1007 structure-specific nano-LC/MS 979:1007 structure-specific nano-LC/MS 979:1007 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 3 26 theme specific 671:678 arg1 regions 680:686 specific regions 671:686 specific regions of the brain 671:699 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 3 27 with sections 736:743 arg1 resolution 777:786 a one-dimensional spatial resolution 751:786 a one-dimensional spatial resolution of 20 μm 751:795 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 1 28 theme glycans 190:196 arg1 extraction 176:185 extraction 176:185 extraction of glycans from tissue 176:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 2 29 theme mass 389:392 arg1 MS 394:395 accurate mass MS 380:395 accurate mass MS 380:395 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 0 30 theme mouse 38:42 arg1 glycome 50:56 the mouse brain glycome 34:56 the mouse brain glycome 34:56 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 3 31 theme spatial 769:775 arg1 resolution 777:786 a one-dimensional spatial resolution 751:786 a one-dimensional spatial resolution of 20 μm 751:795 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 5 32 theme glycome 1118:1124 arg1 exploration 1093:1103 qualitative and quantitative exploration 1064:1103 qualitative and quantitative exploration of the brain glycome 1064:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 4 33 theme R 900:900 arg1 > 902:902 R > 0.98 900:907 R > 0.98 900:907 Reproducibility was extraordinarily high (R > 0.98) for both method and instrumental replicates. 25643168 1 34 theme Tissue 104:109 arg1 method 165:170 a highly sensitive MS-compatible method 132:170 a highly sensitive MS-compatible method for extraction of glycans from tissue 132:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 34 theme Tissue 104:109 arg1 TGC 126:128 TGC 126:128 TGC 126:128 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 34 theme Tissue 104:109 arg1 glyco-capture 111:123 Tissue glyco-capture 104:123 Tissue glyco-capture (TGC) 104:129 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 0 35 theme glycome 50:56 arg1 exploration 19:29 Spatially-resolved exploration 0:29 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 0:102 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 1 36 theme mouse 303:307 arg1 glycome 315:321 the mouse brain glycome 299:321 the mouse brain glycome 299:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 5 37 theme brain 1112:1116 arg1 glycome 1118:1124 the brain glycome 1108:1124 the brain glycome 1108:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 1 38 from extraction 176:185 arg1 tissue 203:208 tissue 203:208 tissue 203:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 39 theme glycomic 650:657 arg1 analysis 659:666 glycomic analysis 650:666 glycomic analysis of specific regions of the brain 650:699 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 1 40 theme brain 309:313 arg1 glycome 315:321 the mouse brain glycome 299:321 the mouse brain glycome 299:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 41 from tissue 203:208 arg1 glycans 190:196 glycans 190:196 glycans from tissue 190:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 1 41 from tissue 203:208 arg1 extraction 176:185 extraction 176:185 extraction of glycans from tissue 176:208 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 42 theme brain 695:699 arg1 regions 680:686 specific regions 671:686 specific regions of the brain 671:699 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 3 43 theme regions 680:686 arg1 analysis 659:666 glycomic analysis 650:666 glycomic analysis of specific regions of the brain 650:699 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 5 44 theme TGC 970:972 arg1 platform 1051:1058 an exceptionally powerful platform 1025:1058 an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome 1025:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 5 44 theme TGC 970:972 arg1 pairing 959:965 The pairing 955:965 The pairing of TGC with structure-specific nano-LC/MS 955:1007 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 5 45 theme powerful 1042:1049 arg1 platform 1051:1058 an exceptionally powerful platform 1025:1058 an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome 1025:1124 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 5 45 theme powerful 1042:1049 arg1 pairing 959:965 The pairing 955:965 The pairing of TGC with structure-specific nano-LC/MS 955:1007 The pairing of TGC with structure-specific nano-LC/MS was found to be an exceptionally powerful platform for qualitative and quantitative exploration of the brain glycome. 25643168 2 46 theme accurate 380:387 arg1 mass 389:392 accurate mass 380:392 accurate mass MS 380:395 Hundreds of glycan structures were directly detected by accurate mass MS and structurally elucidated by MS/MS, revealing the presence of novel glycan motifs such as antennary fucosylation, sulfation, and glucuronidation that are potentially associated with cellular signaling and adhesion. 25643168 3 47 theme μm 794:795 arg1 resolution 777:786 a one-dimensional spatial resolution 751:786 a one-dimensional spatial resolution of 20 μm 751:795 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 25643168 0 48 theme tissue 61:66 arg1 TGC 83:85 TGC 83:85 TGC 83:85 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 0 48 theme tissue 61:66 arg1 glyco-capture 68:80 tissue glyco-capture 61:80 tissue glyco-capture (TGC) 61:86 Spatially-resolved exploration of the mouse brain glycome by tissue glyco-capture (TGC) and nano-LC/MS. 25643168 1 49 theme glycome 315:321 arg1 profiling 286:294 sensitive and detailed profiling 263:294 sensitive and detailed profiling of the mouse brain glycome 263:321 Tissue glyco-capture (TGC), a highly sensitive MS-compatible method for extraction of glycans from tissue, was combined with structure-specific nano-LC/MS for sensitive and detailed profiling of the mouse brain glycome. 25643168 3 50 theme Microgram-level 614:628 arg1 sensitivity 630:640 Microgram-level sensitivity 614:640 Microgram-level sensitivity 614:640 Microgram-level sensitivity enabled glycomic analysis of specific regions of the brain, as demonstrated on not only brain sections (with a one-dimensional spatial resolution of 20 μm) but also isolated brain structures (e.g., the hippocampus). 24799124 0 0 from core-fucosylation 28:44 arg1 carcinoma 97:105 alcohol-related hepatocellular carcinoma 66:105 alcohol-related hepatocellular carcinoma 66:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 6 1 theme test 1427:1430 arg1 AFP 1405:1407 AFP 1405:1407 AFP (AUC = 0.867) (LR test p = 0.0002) 1405:1442 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 1 theme test 1427:1430 arg1 0.0002 1436:1441 LR test p = 0.0002 1424:1441 LR test p = 0.0002 1424:1441 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 2 2 theme different 400:408 arg1 virus 475:479 hepatitis C virus 463:479 hepatitis C virus 463:479 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 2 2 theme different 400:408 arg1 virus 453:457 hepatitis B virus 441:457 hepatitis B virus 441:457 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 2 2 theme different 400:408 arg1 etiologies 410:419 different etiologies 400:419 different etiologies 400:419 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 2 2 theme different 400:408 arg1 alcohol 432:438 alcohol 432:438 alcohol 432:438 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 2 3 theme hepatitis 441:449 arg1 virus 453:457 hepatitis B virus 441:457 hepatitis B virus 441:457 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 5 4 theme AUC 1124:1126 arg1 value 1128:1132 the highest AUC value 1112:1132 the highest AUC value of 0.838 at site 138 1112:1153 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 5 from site 1146:1149 arg1 value 1128:1132 the highest AUC value 1112:1132 the highest AUC value of 0.838 at site 138 1112:1153 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 6 theme core-fucosylation 911:927 arg1 ratio 929:933 The core-fucosylation ratio 907:933 The core-fucosylation ratio of three of these sites 907:957 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 1 7 theme serum 206:210 arg1 ceruloplasmin 212:224 serum ceruloplasmin 206:224 serum ceruloplasmin 206:224 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 6 8 theme = 1347:1347 arg1 0.022 1349:1353 p = 0.022 1345:1353 p = 0.022 1345:1353 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 7 9 located observed 1577:1584 arg2 change 1528:1533 no significant site-specific change 1499:1533 no significant site-specific change in core-fucosylation of ceruloplasmin 1499:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 7 9 located observed 1577:1584 arg1 diseases 1489:1496 HBV- or HCV-related liver diseases 1463:1496 HBV- or HCV-related liver diseases 1463:1496 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 3 10 theme glycan 609:614 arg1 structure 616:624 the glycan structure 605:624 the glycan structure 605:624 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 5 11 theme alcohol-related 986:1000 arg1 samples 1006:1012 alcohol-related HCC samples 986:1012 alcohol-related HCC samples (sample size = 24) 986:1031 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 11 theme alcohol-related 986:1000 arg1 size 1022:1025 sample size = 24 1015:1030 sample size = 24 1015:1030 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 12 theme sites 953:957 arg1 three 938:942 three 938:942 three 938:942 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 12 theme sites 953:957 arg1 sites 953:957 these sites 947:957 these sites 947:957 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 13 theme HCC 1002:1004 arg1 samples 1006:1012 alcohol-related HCC samples 986:1012 alcohol-related HCC samples (sample size = 24) 986:1031 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 13 theme HCC 1002:1004 arg1 size 1022:1025 sample size = 24 1015:1030 sample size = 24 1015:1030 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 6 14 dep = 1337:1337 arg1 3.64 1339:1342 3.64 1339:1342 3.64 1339:1342 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 7 15 theme site-specific 1514:1526 arg1 change 1528:1533 no significant site-specific change 1499:1533 no significant site-specific change in core-fucosylation of ceruloplasmin 1499:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 3 16 theme F3 569:570 arg1 digestion 572:580 Endo F3 digestion 564:580 Endo F3 digestion 564:580 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 7 17 theme ceruloplasmin 1559:1571 arg1 core-fucosylation 1538:1554 core-fucosylation 1538:1554 core-fucosylation of ceruloplasmin 1538:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 1 18 theme ceruloplasmin 212:224 arg1 core-fucosylation 185:201 core-fucosylation 185:201 core-fucosylation of serum ceruloplasmin 185:224 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 6 19 theme = 1311:1311 arg1 0.954 1294:1298 0.954 1294:1298 0.954 1294:1298 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 19 theme = 1311:1311 arg1 ORsite138 1301:1309 ORsite138 = 12.26 1301:1317 ORsite138 = 12.26 1301:1317 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 7 20 gly core-fucosylation 1538:1554 arg1 ceruloplasmin 1559:1571 ceruloplasmin 1559:1571 ceruloplasmin 1559:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 6 21 theme alpha-fetoprotein 1236:1252 arg1 value 1260:1264 the alpha-fetoprotein (AFP) value 1232:1264 the alpha-fetoprotein (AFP) value 1232:1264 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 1 22 theme mass 110:113 arg1 methodology 134:144 A mass spectrometry-based methodology 108:144 A mass spectrometry-based methodology 108:144 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 0 23 theme site-specific 14:26 arg1 core-fucosylation 28:44 Mass-selected site-specific core-fucosylation 0:44 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma 0:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 4 24 gly core-fucosylated 821:836 arg1 sites 845:849 sites 138, 358, 397, and 762 845:872 sites 138, 358, 397, and 762 845:872 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 24 gly core-fucosylated 821:836 arg1 sites 838:842 four core-fucosylated sites 816:842 four core-fucosylated sites (sites 138, 358, 397, and 762) 816:873 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 1 25 from changes 174:180 arg1 core-fucosylation 185:201 core-fucosylation 185:201 core-fucosylation of serum ceruloplasmin 185:224 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 1 26 theme spectrometry-based 115:132 arg1 methodology 134:144 A mass spectrometry-based methodology 108:144 A mass spectrometry-based methodology 108:144 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 6 27 dep = 1322:1322 arg1 0.017 1324:1328 0.017 1324:1328 0.017 1324:1328 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 0 28 theme Mass-selected 0:12 arg1 core-fucosylation 28:44 Mass-selected site-specific core-fucosylation 0:44 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma 0:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 6 29 theme = 1414:1414 arg1 AUC 1410:1412 AUC = 0.867 1410:1420 AUC = 0.867 1410:1420 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 29 theme = 1414:1414 arg1 AFP 1405:1407 AFP 1405:1407 AFP (AUC = 0.867) (LR test p = 0.0002) 1405:1442 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 2 30 theme C 473:473 arg1 virus 475:479 hepatitis C virus 463:479 hepatitis C virus 463:479 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 6 31 theme core-fucosylation 1175:1191 arg1 ratio 1193:1197 the core-fucosylation ratio 1171:1197 the core-fucosylation ratio of site 138 1171:1209 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 7 32 from change 1528:1533 arg1 core-fucosylation 1538:1554 core-fucosylation 1538:1554 core-fucosylation of ceruloplasmin 1538:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 6 33 theme = 1322:1322 arg1 p 1320:1320 p 1320:1320 p = 0.017 1320:1328 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 2 34 theme hepatitis 463:471 arg1 virus 475:479 hepatitis C virus 463:479 hepatitis C virus 463:479 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 5 35 theme highest 1116:1122 arg1 value 1128:1132 the highest AUC value 1112:1132 the highest AUC value of 0.838 at site 138 1112:1153 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 4 36 theme core-fucosylated 821:836 arg1 sites 845:849 sites 138, 358, 397, and 762 845:872 sites 138, 358, 397, and 762 845:872 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 36 theme core-fucosylated 821:836 arg1 sites 838:842 four core-fucosylated sites 816:842 four core-fucosylated sites (sites 138, 358, 397, and 762) 816:873 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 37 theme mass 791:794 arg1 spectrometry 796:807 mass spectrometry 791:807 mass spectrometry 791:807 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 3 38 theme trypsin 503:509 arg1 digestion 511:519 trypsin digestion 503:519 trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide 503:723 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 6 39 theme p 1345:1345 arg1 0.022 1349:1353 p = 0.022 1345:1353 p = 0.022 1345:1353 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 7 40 theme liver 1483:1487 arg1 diseases 1489:1496 HBV- or HCV-related liver diseases 1463:1496 HBV- or HCV-related liver diseases 1463:1496 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 5 41 theme sample 1088:1093 arg1 samples 1079:1085 alcohol-related cirrhosis samples 1053:1085 alcohol-related cirrhosis samples (sample size = 18) 1053:1104 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 41 theme sample 1088:1093 arg1 size 1095:1098 sample size = 18 1088:1103 sample size = 18 1088:1103 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 7 42 theme significant 1502:1512 arg1 change 1528:1533 no significant site-specific change 1499:1533 no significant site-specific change in core-fucosylation of ceruloplasmin 1499:1571 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 0 43 theme ceruloplasmin 49:61 arg1 core-fucosylation 28:44 Mass-selected site-specific core-fucosylation 0:44 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma 0:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 7 44 theme HBV- 1463:1466 arg1 diseases 1489:1496 HBV- or HCV-related liver diseases 1463:1496 HBV- or HCV-related liver diseases 1463:1496 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 1 45 theme hepatocellular 271:284 arg1 HCC 297:299 HCC 297:299 HCC 297:299 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 1 45 theme hepatocellular 271:284 arg1 carcinoma 286:294 hepatocellular carcinoma 271:294 hepatocellular carcinoma (HCC) 271:300 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 0 46 theme hepatocellular 82:95 arg1 carcinoma 97:105 alcohol-related hepatocellular carcinoma 66:105 alcohol-related hepatocellular carcinoma 66:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 5 47 theme 0.838 1137:1141 arg1 value 1128:1132 the highest AUC value 1112:1132 the highest AUC value of 0.838 at site 138 1112:1153 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 1 48 gly core-fucosylation 185:201 arg1 ceruloplasmin 212:224 serum ceruloplasmin 206:224 serum ceruloplasmin 206:224 A mass spectrometry-based methodology has been developed to study changes in core-fucosylation of serum ceruloplasmin that are site-specific between cirrhosis and hepatocellular carcinoma (HCC). 24799124 5 49 theme = 1100:1100 arg1 samples 1079:1085 alcohol-related cirrhosis samples 1053:1085 alcohol-related cirrhosis samples (sample size = 18) 1053:1104 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 49 theme = 1100:1100 arg1 size 1095:1098 sample size = 18 1088:1103 sample size = 18 1088:1103 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 0 50 gly core-fucosylation 28:44 arg1 carcinoma 97:105 alcohol-related hepatocellular carcinoma 66:105 alcohol-related hepatocellular carcinoma 66:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 0 50 gly core-fucosylation 28:44 arg1 ceruloplasmin 49:61 ceruloplasmin 49:61 ceruloplasmin 49:61 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 3 51 theme ceruloplasmin 524:536 arg1 digestion 511:519 trypsin digestion 503:519 trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide 503:723 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 0 52 theme alcohol-related 66:80 arg1 carcinoma 97:105 alcohol-related hepatocellular carcinoma 66:105 alcohol-related hepatocellular carcinoma 66:105 Mass-selected site-specific core-fucosylation of ceruloplasmin in alcohol-related hepatocellular carcinoma. 24799124 7 53 theme HCV-related 1471:1481 arg1 diseases 1489:1496 HBV- or HCV-related liver diseases 1463:1496 HBV- or HCV-related liver diseases 1463:1496 However, in HBV- or HCV-related liver diseases, no significant site-specific change in core-fucosylation of ceruloplasmin was observed between HCC and cirrhosis. 24799124 6 54 theme site 1202:1205 arg1 ratio 1193:1197 the core-fucosylation ratio 1171:1197 the core-fucosylation ratio of site 138 1171:1209 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 5 55 theme three 938:942 arg1 ratio 929:933 The core-fucosylation ratio 907:933 The core-fucosylation ratio of three of these sites 907:957 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 6 56 theme AUC 1271:1273 arg1 value 1275:1279 the AUC value 1267:1279 the AUC value 1267:1279 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 5 57 theme alcohol-related 1053:1067 arg1 samples 1079:1085 alcohol-related cirrhosis samples 1053:1085 alcohol-related cirrhosis samples (sample size = 18) 1053:1104 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 57 theme alcohol-related 1053:1067 arg1 size 1095:1098 sample size = 18 1088:1103 sample size = 18 1088:1103 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 58 theme sample 1015:1020 arg1 samples 1006:1012 alcohol-related HCC samples 986:1012 alcohol-related HCC samples (sample size = 24) 986:1031 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 58 theme sample 1015:1020 arg1 size 1022:1025 sample size = 24 1015:1030 sample size = 24 1015:1030 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 4 59 theme further 771:777 arg1 analysis 779:786 further analysis 771:786 further analysis 771:786 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 60 located detected 880:887 arg2 sites 838:842 four core-fucosylated sites 816:842 four core-fucosylated sites (sites 138, 358, 397, and 762) 816:873 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 60 located detected 880:887 arg2 sites 845:849 sites 138, 358, 397, and 762 845:872 sites 138, 358, 397, and 762 845:872 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 4 60 located detected 880:887 arg1 ceruloplasmin 892:904 ceruloplasmin 892:904 ceruloplasmin 892:904 This procedure simplified the structures for further analysis by mass spectrometry, where four core-fucosylated sites (sites 138, 358, 397, and 762) were detected in ceruloplasmin. 24799124 5 61 theme cirrhosis 1069:1077 arg1 samples 1079:1085 alcohol-related cirrhosis samples 1053:1085 alcohol-related cirrhosis samples (sample size = 18) 1053:1104 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 61 theme cirrhosis 1069:1077 arg1 size 1095:1098 sample size = 18 1088:1103 sample size = 18 1088:1103 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 2 62 theme serum 307:311 arg1 samples 313:319 The serum samples 303:319 The serum samples studied for these changes 303:345 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 2 63 theme B 451:451 arg1 virus 453:457 hepatitis B virus 441:457 hepatitis B virus 441:457 The serum samples studied for these changes were from patients affected by cirrhosis or HCC with different etiologies, including alcohol, hepatitis B virus, or hepatitis C virus. 24799124 5 64 theme = 1027:1027 arg1 samples 1006:1012 alcohol-related HCC samples 986:1012 alcohol-related HCC samples (sample size = 24) 986:1031 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 5 64 theme = 1027:1027 arg1 size 1022:1025 sample size = 24 1015:1030 sample size = 24 1015:1030 The core-fucosylation ratio of three of these sites increased significantly in alcohol-related HCC samples (sample size = 24) compared to that in alcohol-related cirrhosis samples (sample size = 18), with the highest AUC value of 0.838 at site 138. 24799124 6 65 theme = 1434:1434 arg1 AFP 1405:1407 AFP 1405:1407 AFP (AUC = 0.867) (LR test p = 0.0002) 1405:1442 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 65 theme = 1434:1434 arg1 0.0002 1436:1441 LR test p = 0.0002 1424:1441 LR test p = 0.0002 1424:1441 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 66 theme = 1337:1337 arg1 ORAFP 1331:1335 ORAFP 1331:1335 ORAFP = 3.64 1331:1342 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 67 dep ORsite138 1301:1309 arg1 0.022 1349:1353 p = 0.022 1345:1353 p = 0.022 1345:1353 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 67 dep ORsite138 1301:1309 arg1 p 1320:1320 p 1320:1320 p = 0.017 1320:1328 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 67 dep ORsite138 1301:1309 arg1 ORAFP 1331:1335 ORAFP 1331:1335 ORAFP = 3.64 1331:1342 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 3 68 theme innermost 646:654 arg1 GlcNAc 677:682 GlcNAc 677:682 GlcNAc 677:682 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 3 68 theme innermost 646:654 arg1 N-acetylglucosamine 656:674 the innermost N-acetylglucosamine 642:674 the innermost N-acetylglucosamine (GlcNAc) 642:683 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 24799124 6 69 theme p 1432:1432 arg1 AFP 1405:1407 AFP 1405:1407 AFP (AUC = 0.867) (LR test p = 0.0002) 1405:1442 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 6 69 theme p 1432:1432 arg1 0.0002 1436:1441 LR test p = 0.0002 1424:1441 LR test p = 0.0002 1424:1441 When combining the core-fucosylation ratio of site 138 in ceruloplasmin and the alpha-fetoprotein (AFP) value, the AUC value increased to 0.954 (ORsite138 = 12.26, p = 0.017; ORAFP = 3.64, p = 0.022), which was markedly improved compared to that of AFP (AUC = 0.867) (LR test p = 0.0002) alone. 24799124 3 70 theme Endo 564:567 arg1 digestion 572:580 Endo F3 digestion 564:580 Endo F3 digestion 564:580 The methods involved trypsin digestion of ceruloplasmin into peptides followed by Endo F3 digestion, which removed most of the glycan structure while retaining the innermost N-acetylglucosamine (GlcNAc) and/or core-fucose bound to the peptide. 26791533 5 0 theme glycan 753:758 arg1 compositions 760:771 glycan compositions 753:771 glycan compositions 753:771 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 4 1 theme proteins 662:669 arg1 mixture 633:639 a complex, unknown mixture 614:639 a complex, unknown mixture of protease-digested proteins 614:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 7 2 from sites 1017:1021 arg1 glycoproteins 1029:1041 50 glycoproteins 1026:1041 50 glycoproteins 1026:1041 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 6 3 theme seminal 864:870 arg1 glycoproteins 885:897 human seminal plasma total glycoproteins 858:897 human seminal plasma total glycoproteins 858:897 This strategy has been applied to human seminal plasma total glycoproteins. 26791533 7 4 gly glycopeptides 971:983 arg2 glycopeptides 971:983 243 glycopeptides 967:983 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins 967:1041 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 5 5 theme compositions 760:771 arg1 identification 695:708 identification 695:708 identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures 695:796 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 2 6 attach present 320:326 arg2 y 302:302 y 302:302 y 302:302 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 6 attach present 320:326 arg1 glycoproteins 346:358 seminal plasma glycoproteins 331:358 seminal plasma glycoproteins 331:358 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 6 attach present 320:326 arg2 x 296:296 Lewis x 290:296 Lewis x 290:296 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 6 attach present 320:326 arg2 Epitopes 273:280 Epitopes 273:280 Epitopes such as Lewis x and y 273:302 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 8 7 theme structures 1080:1089 arg1 majority 1048:1055 The majority 1044:1055 The majority of the proposed glycan structures 1044:1089 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 8 7 theme structures 1080:1089 arg1 type 1104:1107 complex type 1096:1107 complex type (83%) followed by high-mannose (10%) and then hybrid (7%) 1096:1165 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 10 8 theme seminal 1350:1356 arg1 glycoproteins 1365:1377 multiple seminal plasma glycoproteins 1341:1377 multiple seminal plasma glycoproteins 1341:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 10 9 theme multiple 1341:1348 arg1 glycoproteins 1365:1377 multiple seminal plasma glycoproteins 1341:1377 multiple seminal plasma glycoproteins 1341:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 11 10 theme sample 1580:1585 arg1 complexity 1562:1571 complexity 1562:1571 complexity 1562:1571 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 11 10 theme sample 1580:1585 arg1 prefractionation 1541:1556 prefractionation 1541:1556 prefractionation 1541:1556 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 1 11 theme fertilization 250:262 arg1 process 264:270 the fertilization process 246:270 the fertilization process 246:270 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 5 12 gly glycoproteins 717:729 arg1 glycoproteins 717:729 the glycoproteins 713:729 the glycoproteins 713:729 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 10 13 theme Many 1240:1243 arg1 x/a 1251:1253 Many Lewis x/a and y/b epitopes 1240:1270 x/a 1251:1253 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 3 14 theme seminal 451:457 arg1 glycoproteins 466:478 seminal plasma glycoproteins 451:478 seminal plasma glycoproteins 451:478 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 6 15 theme total 879:883 arg1 glycoproteins 885:897 human seminal plasma total glycoproteins 858:897 human seminal plasma total glycoproteins 858:897 This strategy has been applied to human seminal plasma total glycoproteins. 26791533 4 16 gly glycopeptides 595:607 arg2 glycopeptides 595:607 glycopeptides 595:607 glycopeptides from a complex, unknown mixture of protease-digested proteins 595:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 4 16 gly glycopeptides 595:607 arg1 mixture 633:639 a complex, unknown mixture 614:639 a complex, unknown mixture of protease-digested proteins 614:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 9 17 gly sialylated 1206:1215 arg1 Most 1168:1171 Most 1168:1171 Most 1168:1171 Most of the glycoproteins were either sialylated, fucosylated, or both. 26791533 9 17 gly sialylated 1206:1215 arg1 glycoproteins 1180:1192 the glycoproteins 1176:1192 the glycoproteins 1176:1192 Most of the glycoproteins were either sialylated, fucosylated, or both. 26791533 4 18 from mixture 633:639 arg1 glycopeptides 595:607 glycopeptides 595:607 glycopeptides from a complex, unknown mixture of protease-digested proteins 595:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 9 19 gly glycoproteins 1180:1192 arg1 glycoproteins 1180:1192 the glycoproteins 1176:1192 the glycoproteins 1176:1192 Most of the glycoproteins were either sialylated, fucosylated, or both. 26791533 5 20 theme glycosylation 732:744 arg1 sites 746:750 glycosylation sites 732:750 glycosylation sites 732:750 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 6 21 theme plasma 872:877 arg1 glycoproteins 885:897 human seminal plasma total glycoproteins 858:897 human seminal plasma total glycoproteins 858:897 This strategy has been applied to human seminal plasma total glycoproteins. 26791533 3 22 theme important 508:516 arg1 roles 518:522 important roles 508:522 important roles 508:522 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 1 23 theme oviductal 180:188 arg1 reservoir 196:204 an oviductal sperm reservoir 177:204 an oviductal sperm reservoir 177:204 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 11 24 dep prefractionation 1541:1556 arg1 the 1537:1539 the 1537:1539 the 1537:1539 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 3 25 theme undiscovered 494:505 arg1 roles 518:522 important roles 508:522 important roles 508:522 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 2 26 gly glycoproteins 346:358 arg1 glycoproteins 346:358 seminal plasma glycoproteins 331:358 seminal plasma glycoproteins 331:358 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 7 27 theme proposed 943:950 arg1 structures 952:961 proposed structures 943:961 proposed structures 943:961 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 3 28 gly glycoproteins 466:478 arg1 glycoproteins 466:478 seminal plasma glycoproteins 451:478 seminal plasma glycoproteins 451:478 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 11 29 with achievable 1445:1454 arg1 techniques 1469:1478 current techniques 1461:1478 current techniques 1461:1478 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 1 30 theme sperm 190:194 arg1 reservoir 196:204 an oviductal sperm reservoir 177:204 an oviductal sperm reservoir 177:204 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 2 31 located present 320:326 arg2 y 302:302 y 302:302 y 302:302 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 31 located present 320:326 arg1 glycoproteins 346:358 seminal plasma glycoproteins 331:358 seminal plasma glycoproteins 331:358 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 31 located present 320:326 arg2 x 296:296 Lewis x 290:296 Lewis x 290:296 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 2 31 located present 320:326 arg2 Epitopes 273:280 Epitopes 273:280 Epitopes such as Lewis x and y 273:302 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 7 32 gly N-glycosylation 1001:1015 arg2 sites 1017:1021 73 N-glycosylation sites 998:1021 73 N-glycosylation sites on 50 glycoproteins 998:1041 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 7 32 gly N-glycosylation 1001:1015 arg2 73 998:999 73 998:999 73 998:999 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 8 33 theme glycan 1073:1078 arg1 structures 1080:1089 the proposed glycan structures 1060:1089 the proposed glycan structures 1060:1089 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 7 34 theme glycan 919:924 arg1 compositions 926:937 glycan compositions 919:937 glycan compositions 919:937 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 1 35 theme premature 97:105 arg1 capacitation 107:118 premature capacitation 97:118 premature capacitation 97:118 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 10 36 theme y/b 1259:1261 arg1 epitopes 1263:1270 Many Lewis x/a and y/b epitopes 1240:1270 epitopes 1263:1270 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 1 37 theme reservoir 196:204 arg1 formation 164:172 formation 164:172 formation 164:172 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 1 37 theme reservoir 196:204 arg1 transport 150:158 transport 150:158 transport 150:158 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 0 38 theme Seminal 27:33 arg1 Glycoproteins 42:54 Human Seminal Plasma Glycoproteins 21:54 Human Seminal Plasma Glycoproteins 21:54 N-Glycoproteomics of Human Seminal Plasma Glycoproteins. 26791533 8 39 theme complex 1096:1102 arg1 type 1104:1107 complex type 1096:1107 complex type (83%) followed by high-mannose (10%) and then hybrid (7%) 1096:1165 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 8 39 theme complex 1096:1102 arg1 majority 1048:1055 The majority 1044:1055 The majority of the proposed glycan structures 1044:1089 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 8 39 theme complex 1096:1102 arg1 % 1112:1112 83% 1110:1112 83% 1110:1112 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 10 40 theme plasma 1358:1363 arg1 glycoproteins 1365:1377 multiple seminal plasma glycoproteins 1341:1377 multiple seminal plasma glycoproteins 1341:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 5 41 gly glycosylation 732:744 arg2 sites 746:750 glycosylation sites 732:750 glycosylation sites 732:750 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 11 42 theme analysis 1501:1508 arg1 proportional 1521:1532 proportional 1521:1532 proportional 1521:1532 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 11 42 theme analysis 1501:1508 arg1 depth 1488:1492 the depth 1484:1492 the depth of the analysis 1484:1508 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 2 43 theme immune 393:398 arg1 response 400:407 the maternal immune response 380:407 the maternal immune response 380:407 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 0 44 theme Human 21:25 arg1 Glycoproteins 42:54 Human Seminal Plasma Glycoproteins 21:54 Human Seminal Plasma Glycoproteins 21:54 N-Glycoproteomics of Human Seminal Plasma Glycoproteins. 26791533 1 45 from process 264:270 arg1 important 233:241 important 233:241 important 233:241 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 6 46 theme human 858:862 arg1 glycoproteins 885:897 human seminal plasma total glycoproteins 858:897 human seminal plasma total glycoproteins 858:897 This strategy has been applied to human seminal plasma total glycoproteins. 26791533 2 47 theme Lewis 290:294 arg1 x 296:296 Lewis x 290:296 Lewis x 290:296 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 11 48 theme large 1406:1410 arg1 mapping 1434:1440 large scale N-glycosylation mapping 1406:1440 large scale N-glycosylation mapping 1406:1440 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 2 49 theme maternal 384:391 arg1 response 400:407 the maternal immune response 380:407 the maternal immune response 380:407 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 0 50 theme Glycoproteins 42:54 arg1 N-Glycoproteomics 0:16 N-Glycoproteomics 0:16 N-Glycoproteomics of Human Seminal Plasma Glycoproteins. 0:55 N-Glycoproteomics of Human Seminal Plasma Glycoproteins. 26791533 4 51 theme complex 616:622 arg1 mixture 633:639 a complex, unknown mixture 614:639 a complex, unknown mixture of protease-digested proteins 614:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 4 52 dep complex 616:622 arg1 unknown 625:631 unknown 625:631 unknown 625:631 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 10 53 theme Lewis 1245:1249 arg1 x/a 1251:1253 Many Lewis x/a and y/b epitopes 1240:1270 x/a 1251:1253 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 3 54 theme fertilization 542:554 arg1 process 531:537 the process 527:537 the process of fertilization 527:554 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 0 55 theme Plasma 35:40 arg1 Glycoproteins 42:54 Human Seminal Plasma Glycoproteins 21:54 Human Seminal Plasma Glycoproteins 21:54 N-Glycoproteomics of Human Seminal Plasma Glycoproteins. 26791533 7 56 gly glycoproteins 1029:1041 arg1 glycoproteins 1029:1041 50 glycoproteins 1026:1041 50 glycoproteins 1026:1041 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 8 57 theme proposed 1064:1071 arg1 structures 1080:1089 the proposed glycan structures 1060:1089 the proposed glycan structures 1060:1089 The majority of the proposed glycan structures were complex type (83%) followed by high-mannose (10%) and then hybrid (7%). 26791533 5 58 theme original 807:814 arg1 sample 816:821 the original sample 803:821 the original sample 803:821 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 9 59 gly fucosylated 1218:1228 arg1 Most 1168:1171 Most 1168:1171 Most 1168:1171 Most of the glycoproteins were either sialylated, fucosylated, or both. 26791533 9 59 gly fucosylated 1218:1228 arg1 glycoproteins 1180:1192 the glycoproteins 1176:1192 the glycoproteins 1176:1192 Most of the glycoproteins were either sialylated, fucosylated, or both. 26791533 5 60 theme proposed 778:785 arg1 structures 787:796 proposed structures 778:796 proposed structures 778:796 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 11 61 theme current 1461:1467 arg1 techniques 1469:1478 current techniques 1461:1478 current techniques 1461:1478 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 3 62 theme plasma 459:464 arg1 glycoproteins 466:478 seminal plasma glycoproteins 451:478 seminal plasma glycoproteins 451:478 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 5 63 theme structures 787:796 arg1 identification 695:708 identification 695:708 identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures 695:796 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 1 64 from important 233:241 arg1 process 264:270 the fertilization process 246:270 the fertilization process 246:270 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 10 65 from epitopes 1329:1336 arg1 glycoproteins 1365:1377 multiple seminal plasma glycoproteins 1341:1377 multiple seminal plasma glycoproteins 1341:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 7 66 theme N-glycosylation 1001:1015 arg1 sites 1017:1021 73 N-glycosylation sites 998:1021 73 N-glycosylation sites on 50 glycoproteins 998:1041 We have elucidated glycan compositions and proposed structures for 243 glycopeptides belonging to 73 N-glycosylation sites on 50 glycoproteins. 26791533 4 67 theme protease-digested 644:660 arg1 proteins 662:669 protease-digested proteins 644:669 protease-digested proteins 644:669 We have devised a strategy to analyze glycopeptides from a complex, unknown mixture of protease-digested proteins. 26791533 6 68 gly glycoproteins 885:897 arg1 glycoproteins 885:897 human seminal plasma total glycoproteins 858:897 human seminal plasma total glycoproteins 858:897 This strategy has been applied to human seminal plasma total glycoproteins. 26791533 10 69 theme immune-modulating 1311:1327 arg1 epitopes 1329:1336 immune-modulating epitopes 1311:1336 immune-modulating epitopes on multiple seminal plasma glycoproteins 1311:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 1 70 theme intracervical 136:148 arg1 transport 150:158 transport 150:158 transport 150:158 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 5 71 theme sites 746:750 arg1 identification 695:708 identification 695:708 identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures 695:796 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 26791533 3 72 theme multiple 429:436 arg1 studies 438:444 multiple studies 429:444 multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization 429:554 It is suggested by multiple studies that seminal plasma glycoproteins play, largely undiscovered, important roles in the process of fertilization. 26791533 2 73 theme plasma 339:344 arg1 glycoproteins 346:358 seminal plasma glycoproteins 331:358 seminal plasma glycoproteins 331:358 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 11 74 theme N-glycosylation 1418:1432 arg1 mapping 1434:1440 large scale N-glycosylation mapping 1406:1440 large scale N-glycosylation mapping 1406:1440 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 11 75 theme scale 1412:1416 arg1 mapping 1434:1440 large scale N-glycosylation mapping 1406:1440 large scale N-glycosylation mapping 1406:1440 The study also shows that large scale N-glycosylation mapping is achievable with current techniques and the depth of the analysis is roughly proportional to the prefractionation and complexity of the sample. 26791533 10 76 gly glycoproteins 1365:1377 arg1 glycoproteins 1365:1377 multiple seminal plasma glycoproteins 1341:1377 multiple seminal plasma glycoproteins 1341:1377 Many Lewis x/a and y/b epitopes bearing glycans were found, suggesting immune-modulating epitopes on multiple seminal plasma glycoproteins. 26791533 1 77 theme Seminal 57:63 arg1 plasma 65:70 Seminal plasma 57:70 Seminal plasma 57:70 Seminal plasma aids sperm by inhibiting premature capacitation, helping in the intracervical transport and formation of an oviductal sperm reservoir, all of which appear to be important in the fertilization process. 26791533 2 78 theme seminal 331:337 arg1 glycoproteins 346:358 seminal plasma glycoproteins 331:358 seminal plasma glycoproteins 331:358 Epitopes such as Lewis x and y are known to be present on seminal plasma glycoproteins, which can modulate the maternal immune response. 26791533 5 79 theme glycoproteins 717:729 arg1 identification 695:708 identification 695:708 identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures 695:796 This analysis provides identification of the glycoproteins, glycosylation sites, glycan compositions, and proposed structures from the original sample. 25737449 0 0 theme virus 69:73 arg1 glycoprotein 78:89 hepatitis C virus E2 glycoprotein 57:89 hepatitis C virus E2 glycoprotein 57:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 10 1 theme evasion 1650:1656 arg1 epitope 1612:1618 the E2(412-423) epitope 1596:1618 the E2(412-423) epitope 1596:1618 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 10 1 theme evasion 1650:1656 arg1 strategy 1658:1665 an immune evasion strategy 1640:1665 an immune evasion strategy 1640:1665 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 9 2 theme Asn-415 1487:1493 arg1 glycosylation 1470:1482 glycosylation 1470:1482 glycosylation of Asn-415 1470:1493 Whereas Asn-415 is buried by HCV1 and AP33, it is solvent-exposed in the HC33.1-E2(412-423) complex, such that glycosylation of Asn-415 would not prevent antibody binding. 25737449 1 3 theme C 143:143 arg1 cause 168:172 a major cause 160:172 a major cause of liver cirrhosis and hepatocellular carcinoma 160:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 3 theme C 143:143 arg1 HCV 152:154 HCV 152:154 HCV 152:154 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 3 theme C 143:143 arg1 virus 145:149 Hepatitis C virus 133:149 Hepatitis C virus (HCV) 133:155 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 4 theme carcinoma 212:220 arg1 cause 168:172 a major cause 160:172 a major cause of liver cirrhosis and hepatocellular carcinoma 160:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 4 theme carcinoma 212:220 arg1 virus 145:149 Hepatitis C virus 133:149 Hepatitis C virus (HCV) 133:155 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 0 5 theme C 67:67 arg1 glycoprotein 78:89 hepatitis C virus E2 glycoprotein 57:89 hepatitis C virus E2 glycoprotein 57:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 2 6 theme protective 312:321 arg1 antibodies 323:332 protective antibodies 312:332 protective antibodies against this highly diverse virus 312:366 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 10 7 theme antibodies 1690:1699 arg1 induction 1677:1685 induction 1677:1685 induction of antibodies targeting this site 1677:1719 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 0 8 theme glycoprotein 78:89 arg1 glycan 40:45 the glycan shield 36:52 the glycan shield of hepatitis C virus E2 glycoprotein 36:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 9 9 theme antibody 1513:1520 arg1 binding 1522:1528 antibody binding 1513:1528 antibody binding 1513:1528 Whereas Asn-415 is buried by HCV1 and AP33, it is solvent-exposed in the HC33.1-E2(412-423) complex, such that glycosylation of Asn-415 would not prevent antibody binding. 25737449 7 10 from structure 976:984 arg1 complex 1022:1028 complex 1022:1028 complex with its E2(412-423) epitope to 2.0 Å resolution 1022:1077 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 0 11 theme E2 75:76 arg1 glycoprotein 78:89 hepatitis C virus E2 glycoprotein 57:89 hepatitis C virus E2 glycoprotein 57:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 5 12 theme linear 613:618 arg1 epitope 620:626 this linear epitope 608:626 this linear epitope 608:626 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 8 13 from conformation 1159:1170 arg1 distinct 1131:1138 distinct 1131:1138 distinct 1131:1138 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 8 13 from conformation 1159:1170 arg1 conformation 1084:1095 The conformation 1080:1095 The conformation of E2(412-423) bound to HC33.1 1080:1126 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 2 14 theme vaccine 243:249 arg1 development 251:261 HCV vaccine development 239:261 HCV vaccine development 239:261 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 3 15 theme E2 441:442 arg1 glycoprotein 453:464 its E2 envelope glycoprotein 437:464 its E2 envelope glycoprotein 437:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 7 16 theme neutralizing 1002:1013 arg1 mAb 1015:1017 this broadly neutralizing mAb 989:1017 this broadly neutralizing mAb 989:1017 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 2 17 theme HCV 239:241 arg1 development 251:261 HCV vaccine development 239:261 HCV vaccine development 239:261 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 8 18 theme chain 1352:1356 arg1 region 1323:1328 the unusually long complementarity-determining region 3 1276:1330 the unusually long complementarity-determining region 3 of the HC33.1 heavy chain 1276:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 6 19 theme mAb 797:799 arg1 HC33.1 801:806 the human mAb HC33.1 787:806 the human mAb HC33.1 787:806 By contrast, the human mAb HC33.1 can neutralize virus bearing the N417S mutation. 25737449 7 20 theme mAb 1015:1017 arg1 structure 976:984 the structure 972:984 the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution 972:1077 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 7 21 theme Å 1066:1066 arg1 resolution 1068:1077 2.0 Å resolution 1062:1077 2.0 Å resolution 1062:1077 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 4 22 theme neutralizing 550:561 arg1 activities 563:572 broadly neutralizing activities 542:572 broadly neutralizing activities 542:572 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 6 23 theme human 791:795 arg1 HC33.1 801:806 the human mAb HC33.1 787:806 the human mAb HC33.1 787:806 By contrast, the human mAb HC33.1 can neutralize virus bearing the N417S mutation. 25737449 2 24 theme conserved 278:286 arg1 epitopes 288:295 conserved epitopes 278:295 conserved epitopes able to elicit protective antibodies against this highly diverse virus 278:366 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 2 25 theme diverse 354:360 arg1 virus 362:366 this highly diverse virus 342:366 this highly diverse virus 342:366 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 0 26 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 0:131 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 5 27 theme adaptive 587:594 arg1 N417S 629:633 N417S 629:633 N417S 629:633 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 5 27 theme adaptive 587:594 arg1 mutation 596:603 an adaptive mutation 584:603 an adaptive mutation in this linear epitope 584:626 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 0 28 gly glycoprotein 78:89 arg1 glycoprotein 78:89 hepatitis C virus E2 glycoprotein 57:89 hepatitis C virus E2 glycoprotein 57:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 5 29 theme glycosylation 657:669 arg1 shift 671:675 a glycosylation shift 655:675 a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33 655:771 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 10 30 theme immune 1643:1648 arg1 epitope 1612:1618 the E2(412-423) epitope 1596:1618 the E2(412-423) epitope 1596:1618 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 10 30 theme immune 1643:1648 arg1 strategy 1658:1665 an immune evasion strategy 1640:1665 an immune evasion strategy 1640:1665 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 8 31 with interactions 1258:1269 arg1 region 1323:1328 the unusually long complementarity-determining region 3 1276:1330 the unusually long complementarity-determining region 3 of the HC33.1 heavy chain 1276:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 0 32 theme neutralizing 104:115 arg1 antibody 123:130 a broadly neutralizing human antibody 94:130 a broadly neutralizing human antibody 94:130 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 9 33 theme HC33.1-E2 1432:1440 arg1 complex 1451:1457 the HC33.1-E2(412-423) complex 1428:1457 the HC33.1-E2(412-423) complex 1428:1457 Whereas Asn-415 is buried by HCV1 and AP33, it is solvent-exposed in the HC33.1-E2(412-423) complex, such that glycosylation of Asn-415 would not prevent antibody binding. 25737449 7 34 dep glycan 897:902 arg1 shield 904:909 shield 904:909 shield 904:909 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 0 35 theme broadly 96:102 arg1 antibody 123:130 a broadly neutralizing human antibody 94:130 a broadly neutralizing human antibody 94:130 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 8 36 theme β-hairpin 1240:1248 arg1 disruption 1222:1231 disruption 1222:1231 disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain 1222:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 1 37 theme major 162:166 arg1 cause 168:172 a major cause 160:172 a major cause of liver cirrhosis and hepatocellular carcinoma 160:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 37 theme major 162:166 arg1 virus 145:149 Hepatitis C virus 133:149 Hepatitis C virus (HCV) 133:155 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 8 38 theme E2 1100:1101 arg1 distinct 1131:1138 distinct 1131:1138 distinct 1131:1138 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 8 38 theme E2 1100:1101 arg1 conformation 1084:1095 The conformation 1080:1095 The conformation of E2(412-423) bound to HC33.1 1080:1126 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 5 39 from Asn-417 682:688 arg1 shift 671:675 a glycosylation shift 655:675 a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33 655:771 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 0 40 theme human 117:121 arg1 antibody 123:130 a broadly neutralizing human antibody 94:130 a broadly neutralizing human antibody 94:130 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 8 41 theme peptide 1180:1186 arg1 conformation 1159:1170 the β-hairpin conformation 1145:1170 the β-hairpin conformation of this peptide bound to HCV1 or AP33 1145:1208 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 0 42 dep glycan 40:45 arg1 shield 47:52 shield 47:52 shield 47:52 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 7 43 theme glycosylation 926:938 arg1 shift 940:944 the glycosylation shift 922:944 the glycosylation shift to Asn-415 922:955 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 3 44 theme such 420:423 arg1 epitopes 425:432 such epitopes 420:432 such epitopes on its E2 envelope glycoprotein 420:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 3 45 theme Glycan 369:374 arg1 shielding 376:384 Glycan shielding 369:384 Glycan shielding 369:384 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 3 45 theme Glycan 369:374 arg1 mechanism 391:399 a mechanism 389:399 a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein 389:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 4 46 theme E2 485:486 arg1 region 488:493 the E2 region 481:493 the E2 region comprising residues 412-423 (E2(412-423)) 481:535 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 3 47 theme envelope 444:451 arg1 glycoprotein 453:464 its E2 envelope glycoprotein 437:464 its E2 envelope glycoprotein 437:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 0 48 theme glycan 40:45 arg1 penetration 21:31 penetration 21:31 penetration of the glycan shield of hepatitis C virus E2 glycoprotein 21:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 5 49 from mutation 596:603 arg1 epitope 620:626 this linear epitope 608:626 this linear epitope 608:626 However, an adaptive mutation in this linear epitope, N417S, is associated with a glycosylation shift from Asn-417 to Asn-415 that enables HCV to escape neutralization by mAbs such as HCV1 and AP33. 25737449 9 50 dep prevent 1505:1511 arg1 such 1460:1463 such 1460:1463 such 1460:1463 Whereas Asn-415 is buried by HCV1 and AP33, it is solvent-exposed in the HC33.1-E2(412-423) complex, such that glycosylation of Asn-415 would not prevent antibody binding. 25737449 8 51 theme heavy 1346:1350 arg1 chain 1352:1356 the HC33.1 heavy chain 1335:1356 the HC33.1 heavy chain 1335:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 10 52 theme E2 1600:1601 arg1 epitope 1612:1618 the E2(412-423) epitope 1596:1618 the E2(412-423) epitope 1596:1618 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 10 52 theme E2 1600:1601 arg1 strategy 1658:1665 an immune evasion strategy 1640:1665 an immune evasion strategy 1640:1665 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 3 53 from epitopes 425:432 arg1 glycoprotein 453:464 its E2 envelope glycoprotein 437:464 its E2 envelope glycoprotein 437:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 8 54 theme β-hairpin 1149:1157 arg1 conformation 1159:1170 the β-hairpin conformation 1145:1170 the β-hairpin conformation of this peptide bound to HCV1 or AP33 1145:1208 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 6 55 theme N417S 841:845 arg1 mutation 847:854 the N417S mutation 837:854 the N417S mutation 837:854 By contrast, the human mAb HC33.1 can neutralize virus bearing the N417S mutation. 25737449 8 56 theme HC33.1 1339:1344 arg1 chain 1352:1356 the HC33.1 heavy chain 1335:1356 the HC33.1 heavy chain 1335:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 4 57 contain have 537:540 arg1 Antibodies 467:476 Antibodies 467:476 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) 467:535 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 4 57 contain have 537:540 arg2 activities 563:572 broadly neutralizing activities 542:572 broadly neutralizing activities 542:572 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 2 58 theme able 297:300 arg1 epitopes 288:295 conserved epitopes 278:295 conserved epitopes able to elicit protective antibodies against this highly diverse virus 278:366 A challenge for HCV vaccine development is to identify conserved epitopes able to elicit protective antibodies against this highly diverse virus. 25737449 10 59 theme epitope 1612:1618 arg1 flexibility 1581:1591 the structural flexibility 1566:1591 the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity 1566:1748 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 1 60 theme liver 177:181 arg1 cirrhosis 183:191 liver cirrhosis 177:191 liver cirrhosis 177:191 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 3 61 gly glycoprotein 453:464 arg1 glycoprotein 453:464 its E2 envelope glycoprotein 437:464 its E2 envelope glycoprotein 437:464 Glycan shielding is a mechanism by which HCV masks such epitopes on its E2 envelope glycoprotein. 25737449 1 62 theme cirrhosis 183:191 arg1 cause 168:172 a major cause 160:172 a major cause of liver cirrhosis and hepatocellular carcinoma 160:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 62 theme cirrhosis 183:191 arg1 virus 145:149 Hepatitis C virus 133:149 Hepatitis C virus (HCV) 133:155 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 0 63 theme hepatitis 57:65 arg1 glycoprotein 78:89 hepatitis C virus E2 glycoprotein 57:89 hepatitis C virus E2 glycoprotein 57:89 Structural basis for penetration of the glycan shield of hepatitis C virus E2 glycoprotein by a broadly neutralizing human antibody. 25737449 7 64 with complex 1022:1028 arg1 epitope 1051:1057 its E2(412-423) epitope 1035:1057 its E2(412-423) epitope to 2.0 Å resolution 1035:1077 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 8 65 theme complementarity-determining 1295:1321 arg1 region 1323:1328 the unusually long complementarity-determining region 3 1276:1330 the unusually long complementarity-determining region 3 of the HC33.1 heavy chain 1276:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 9 66 gly glycosylation 1470:1482 arg1 Asn-415 1487:1493 Asn-415 1487:1493 Asn-415 1487:1493 Whereas Asn-415 is buried by HCV1 and AP33, it is solvent-exposed in the HC33.1-E2(412-423) complex, such that glycosylation of Asn-415 would not prevent antibody binding. 25737449 10 67 theme structural 1570:1579 arg1 flexibility 1581:1591 the structural flexibility 1566:1591 the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity 1566:1748 Furthermore, our results highlight the structural flexibility of the E2(412-423) epitope, which may serve as an immune evasion strategy to impede induction of antibodies targeting this site by reducing its antigenicity. 25737449 7 68 theme E2 1039:1040 arg1 epitope 1051:1057 its E2(412-423) epitope 1035:1057 its E2(412-423) epitope to 2.0 Å resolution 1035:1077 To understand how HC33.1 penetrates the glycan shield created by the glycosylation shift to Asn-415, we determined the structure of this broadly neutralizing mAb in complex with its E2(412-423) epitope to 2.0 Å resolution. 25737449 4 69 dep residues 506:513 arg1 E2 524:525 E2(412-423) 524:534 E2(412-423) 524:534 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 4 69 dep residues 506:513 arg1 412-423 515:521 412-423 515:521 412-423 515:521 Antibodies to the E2 region comprising residues 412-423 (E2(412-423)) have broadly neutralizing activities. 25737449 8 70 theme long 1290:1293 arg1 region 1323:1328 the unusually long complementarity-determining region 3 1276:1330 the unusually long complementarity-determining region 3 of the HC33.1 heavy chain 1276:1356 The conformation of E2(412-423) bound to HC33.1 is distinct from the β-hairpin conformation of this peptide bound to HCV1 or AP33, because of disruption of the β-hairpin through interactions with the unusually long complementarity-determining region 3 of the HC33.1 heavy chain. 25737449 1 71 theme Hepatitis 133:141 arg1 cause 168:172 a major cause 160:172 a major cause of liver cirrhosis and hepatocellular carcinoma 160:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 71 theme Hepatitis 133:141 arg1 HCV 152:154 HCV 152:154 HCV 152:154 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 71 theme Hepatitis 133:141 arg1 virus 145:149 Hepatitis C virus 133:149 Hepatitis C virus (HCV) 133:155 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 25737449 1 72 theme hepatocellular 197:210 arg1 carcinoma 212:220 hepatocellular carcinoma 197:220 hepatocellular carcinoma 197:220 Hepatitis C virus (HCV) is a major cause of liver cirrhosis and hepatocellular carcinoma. 27604319 4 0 gly glycosylation 713:725 arg1 gp120 730:734 gp120 730:734 gp120 730:734 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 5 1 theme single 1061:1066 arg1 site 1068:1071 a single site 1059:1071 a single site 1059:1071 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 3 2 theme potent 647:652 arg1 antibodies 675:684 the most potent broadly neutralizing antibodies 638:684 the most potent broadly neutralizing antibodies 638:684 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 2 3 theme shield 431:436 arg1 " 437:437 this "glycan shield" 418:437 this "glycan shield" 418:437 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 3 4 theme recognition 615:625 arg1 targets 604:610 the targets 600:610 the targets of recognition by some of the most potent broadly neutralizing antibodies 600:684 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 0 5 theme antibody 109:116 arg1 binding 118:124 broadly neutralizing antibody binding 88:124 broadly neutralizing antibody binding 88:124 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 1 6 theme major 297:301 arg1 target 311:316 a major vaccine target 295:316 a major vaccine target 295:316 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 4 7 theme site-specific 699:711 arg1 glycosylation 713:725 the site-specific glycosylation 695:725 the site-specific glycosylation of gp120 695:734 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 5 8 theme first 932:936 arg1 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 0 9 theme neutralizing 96:107 arg1 binding 118:124 broadly neutralizing antibody binding 88:124 broadly neutralizing antibody binding 88:124 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 1 10 theme vaccine 303:309 arg1 target 311:316 a major vaccine target 295:316 a major vaccine target 295:316 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 11 theme essential 233:241 arg1 role 243:246 an essential role 230:246 an essential role 230:246 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 5 12 attach derived 986:992 arg2 gp120 980:984 gp120 980:984 gp120 derived from virions produced by infected T lymphoid cells 980:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 12 attach derived 986:992 arg1 virions 999:1005 virions 999:1005 virions produced by infected T lymphoid cells 999:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 1 13 theme Human 169:173 arg1 type 198:201 Human immunodeficiency virus type 1 169:203 Human immunodeficiency virus type 1 (HIV-1) 169:211 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 13 theme Human 169:173 arg1 HIV-1 206:210 HIV-1 206:210 HIV-1 206:210 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 13 theme Human 169:173 arg1 gp120 214:218 gp120 214:218 gp120(SU) 214:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 2 14 theme glycan 424:429 arg1 " 437:437 this "glycan shield" 418:437 this "glycan shield" 418:437 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 1 15 theme immunodeficiency 175:190 arg1 type 198:201 Human immunodeficiency virus type 1 169:203 Human immunodeficiency virus type 1 (HIV-1) 169:211 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 15 theme immunodeficiency 175:190 arg1 HIV-1 206:210 HIV-1 206:210 HIV-1 206:210 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 15 theme immunodeficiency 175:190 arg1 gp120 214:218 gp120 214:218 gp120(SU) 214:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 4 16 theme vaccine 804:810 arg1 development 812:822 HIV vaccine development 800:822 HIV vaccine development 800:822 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 5 17 theme recombinant 885:895 arg1 forms 897:901 recombinant forms 885:901 recombinant forms of gp120 885:910 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 1 18 theme virus 192:196 arg1 type 198:201 Human immunodeficiency virus type 1 169:203 Human immunodeficiency virus type 1 (HIV-1) 169:211 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 18 theme virus 192:196 arg1 HIV-1 206:210 HIV-1 206:210 HIV-1 206:210 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 18 theme virus 192:196 arg1 gp120 214:218 gp120 214:218 gp120(SU) 214:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 2 19 theme considerable 391:402 arg1 evidence 404:411 considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization 391:502 considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization 391:502 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 4 20 theme HIV 800:802 arg1 development 812:822 HIV vaccine development 800:822 HIV vaccine development 800:822 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 0 21 link virion-derived 38:51 arg1 gp120 59:63 virion-derived HIV-1 gp120 38:63 virion-derived HIV-1 gp120 38:63 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 5 22 theme forms 897:901 arg1 analysis 873:880 glycan analysis 866:880 glycan analysis of recombinant forms of gp120 866:910 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 1 23 theme type 198:201 arg1 SU 162:163 SU 162:163 SU 162:163 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 23 theme type 198:201 arg1 glycoprotein 148:159 The surface envelope glycoprotein 127:159 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) 127:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 5 24 theme gp120 980:984 arg1 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 3 25 theme recent 508:513 arg1 years 515:519 recent years 508:519 recent years 508:519 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 6 26 theme immunogens 1176:1185 arg1 design 1158:1163 the design 1154:1163 the design of vaccine immunogens 1154:1185 These results should help guide the design of vaccine immunogens. 27604319 0 27 theme complete 12:19 arg1 glycoproteome 21:33 the complete glycoproteome 8:33 the complete glycoproteome of virion-derived HIV-1 gp120 8:63 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 1 28 theme virus 251:255 arg1 binding 257:263 virus binding 251:263 virus binding to target CD4+ T-cells 251:286 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 2 29 contain has 325:327 arg2 levels 345:350 remarkably high levels 329:350 remarkably high levels of N-linked glycosylation 329:376 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 2 29 contain has 325:327 arg1 Gp120 319:323 Gp120 319:323 Gp120 319:323 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 5 30 theme gp120 906:910 arg1 forms 897:901 recombinant forms 885:901 recombinant forms of gp120 885:910 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 3 31 dep potent 647:652 arg1 neutralizing 662:673 neutralizing 662:673 neutralizing 662:673 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 2 32 theme glycosylation 364:376 arg1 levels 345:350 remarkably high levels 329:350 remarkably high levels of N-linked glycosylation 329:376 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 0 33 theme virion-derived 38:51 arg1 gp120 59:63 virion-derived HIV-1 gp120 38:63 virion-derived HIV-1 gp120 38:63 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 5 34 theme glycan 866:871 arg1 analysis 873:880 glycan analysis 866:880 glycan analysis of recombinant forms of gp120 866:910 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 1 35 gly glycoprotein 148:159 arg1 SU 162:163 SU 162:163 SU 162:163 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 35 gly glycoprotein 148:159 arg1 glycoprotein 148:159 The surface envelope glycoprotein 127:159 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) 127:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 5 36 theme infected 1019:1026 arg1 cells 1039:1043 infected T lymphoid cells 1019:1043 infected T lymphoid cells 1019:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 37 theme site 963:966 arg1 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 38 theme most 831:834 arg1 studies 842:848 most prior studies 831:848 most prior studies 831:848 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 2 39 theme N-linked 355:362 arg1 glycosylation 364:376 N-linked glycosylation 355:376 N-linked glycosylation 355:376 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 3 40 gly glycosylation 562:574 arg1 targets 604:610 the targets 600:610 the targets of recognition by some of the most potent broadly neutralizing antibodies 600:684 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 2 41 theme antibody-mediated 471:487 arg1 neutralization 489:502 antibody-mediated neutralization 471:502 antibody-mediated neutralization 471:502 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 5 42 theme T 1028:1028 arg1 cells 1039:1043 infected T lymphoid cells 1019:1043 infected T lymphoid cells 1019:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 4 43 theme rational 755:762 arg1 design 764:769 the rational design 751:769 the rational design of glycopeptide antigens for HIV vaccine development 751:822 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 5 44 theme complex 1105:1111 arg1 glycans 1113:1119 complex glycans 1105:1119 complex glycans 1105:1119 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 0 45 theme gp120 59:63 arg1 glycoproteome 21:33 the complete glycoproteome 8:33 the complete glycoproteome of virion-derived HIV-1 gp120 8:63 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 1 46 theme target 268:273 arg1 T-cells 280:286 target CD4+ T-cells 268:286 target CD4+ T-cells 268:286 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 5 47 theme lymphoid 1030:1037 arg1 cells 1039:1043 infected T lymphoid cells 1019:1043 infected T lymphoid cells 1019:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 4 48 theme glycopeptide 774:785 arg1 antigens 787:794 glycopeptide antigens 774:794 glycopeptide antigens 774:794 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 0 49 theme HIV-1 53:57 arg1 gp120 59:63 virion-derived HIV-1 gp120 38:63 virion-derived HIV-1 gp120 38:63 Mapping the complete glycoproteome of virion-derived HIV-1 gp120 provides insights into broadly neutralizing antibody binding. 27604319 1 50 theme CD4+ 275:278 arg1 T-cells 280:286 target CD4+ T-cells 268:286 target CD4+ T-cells 268:286 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 2 51 link N-linked 355:362 arg1 glycosylation 364:376 N-linked glycosylation 355:376 N-linked glycosylation 355:376 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 4 52 theme antigens 787:794 arg1 design 764:769 the rational design 751:769 the rational design of glycopeptide antigens for HIV vaccine development 751:822 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 1 53 theme surface 131:137 arg1 SU 162:163 SU 162:163 SU 162:163 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 53 theme surface 131:137 arg1 glycoprotein 148:159 The surface envelope glycoprotein 127:159 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) 127:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 5 54 theme glycosylation 949:961 arg1 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 3 55 theme gp120 556:560 arg1 glycosylation 562:574 gp120 glycosylation 556:574 gp120 glycosylation 556:574 In recent years, however, it has become clear that gp120 glycosylation can also be included in the targets of recognition by some of the most potent broadly neutralizing antibodies. 27604319 6 56 theme vaccine 1168:1174 arg1 immunogens 1176:1185 vaccine immunogens 1168:1185 vaccine immunogens 1168:1185 These results should help guide the design of vaccine immunogens. 27604319 1 57 theme envelope 139:146 arg1 SU 162:163 SU 162:163 SU 162:163 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 1 57 theme envelope 139:146 arg1 glycoprotein 148:159 The surface envelope glycoprotein 127:159 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) 127:222 The surface envelope glycoprotein (SU) of Human immunodeficiency virus type 1 (HIV-1), gp120(SU) plays an essential role in virus binding to target CD4+ T-cells and is a major vaccine target. 27604319 4 58 theme gp120 730:734 arg1 glycosylation 713:725 the site-specific glycosylation 695:725 the site-specific glycosylation of gp120 695:734 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 27604319 5 59 theme systematic 938:947 arg1 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 60 theme prior 836:840 arg1 studies 842:848 most prior studies 831:848 most prior studies 831:848 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 2 61 theme high 340:343 arg1 levels 345:350 remarkably high levels 329:350 remarkably high levels of N-linked glycosylation 329:376 Gp120 has remarkably high levels of N-linked glycosylation and there is considerable evidence that this "glycan shield" can help protect the virus from antibody-mediated neutralization. 27604319 5 62 gly glycosylation 949:961 arg2 analysis 968:975 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 62 gly glycosylation 949:961 arg2 site 963:966 the first systematic glycosylation site analysis 928:975 the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells 928:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 5 62 gly glycosylation 949:961 arg1 gp120 980:984 gp120 980:984 gp120 derived from virions produced by infected T lymphoid cells 980:1043 While most prior studies have focused on glycan analysis of recombinant forms of gp120, here we report the first systematic glycosylation site analysis of gp120 derived from virions produced by infected T lymphoid cells and show that a single site is exclusively substituted with complex glycans. 27604319 4 63 gly glycopeptide 774:785 arg2 glycopeptide 774:785 glycopeptide antigens 774:794 glycopeptide antigens 774:794 Knowing the site-specific glycosylation of gp120 can facilitate the rational design of glycopeptide antigens for HIV vaccine development. 29069609 10 0 from mixtures 1732:1739 arg1 glycopeptides 1694:1706 specific glycopeptides 1685:1706 specific glycopeptides from complex biological mixtures 1685:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 12 1 theme immunoglobulins 1953:1967 arg1 analysis 1941:1948 the analysis 1937:1948 the analysis of immunoglobulins 1937:1967 This is demonstrated through the analysis of immunoglobulins in human plasma where we detected two IgG1 glycoforms that are rarely observed. 29069609 10 2 theme complex 1713:1719 arg1 mixtures 1732:1739 complex biological mixtures 1713:1739 complex biological mixtures 1713:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 3 3 theme acquisition 593:603 arg1 acquisition 560:570 data-independent acquisition 543:570 data-independent acquisition (DIA) 543:576 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 3 theme acquisition 593:603 arg1 strategies 605:614 alternative acquisition strategies 581:614 alternative acquisition strategies for glycopeptide analysis 581:640 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 3 theme acquisition 593:603 arg1 pseudo-MRM 519:528 pseudo-MRM 519:528 pseudo-MRM (MRM-HR) 519:537 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 5 4 gly N-glycosylation 922:936 arg2 three 916:920 three 916:920 three 916:920 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 5 4 gly N-glycosylation 922:936 arg2 sites 938:942 all three N-glycosylation sites 912:942 all three N-glycosylation sites 912:942 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 4 5 theme data 723:726 arg1 post-acquisition 740:755 data re-analysis post-acquisition 723:755 data re-analysis post-acquisition 723:755 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 7 6 theme method 1165:1170 arg1 development 1135:1145 development 1135:1145 development of a targeted DIA method 1135:1170 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 9 7 theme truncated 1553:1561 arg1 structures 1563:1572 two truncated structures 1549:1572 two truncated structures that are rarely reported 1549:1597 21 glycoforms of IgG1 glycopeptides were identified including two truncated structures that are rarely reported. 29069609 2 8 gly glycopeptides 302:314 arg2 glycopeptides 302:314 glycopeptides 302:314 glycopeptides from data-dependent acquisition (DDA) 302:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 2 8 gly glycopeptides 302:314 arg1 acquisition 336:346 data-dependent acquisition 321:346 data-dependent acquisition (DDA) 321:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 2 8 gly glycopeptides 302:314 arg1 DDA 349:351 DDA 349:351 DDA 349:351 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 7 9 theme targeted 1152:1159 arg1 method 1165:1170 a targeted DIA method 1150:1170 a targeted DIA method 1150:1170 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 6 10 theme acquisition 1040:1050 arg1 limitation 1019:1028 the duty cycle limitation 1004:1028 the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma 1004:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 4 11 theme MS/MS 697:701 arg1 range 708:712 the full MS/MS scan range 688:712 the full MS/MS scan range allowing data re-analysis post-acquisition 688:755 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 0 12 theme compositional 131:143 arg1 knowledge 145:153 predefined glycan compositional knowledge 113:153 predefined glycan compositional knowledge 113:153 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 5 13 theme N-glycosylation 922:936 arg1 sites 938:942 all three N-glycosylation sites 912:942 all three N-glycosylation sites 912:942 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 7 14 theme glycan 1237:1242 arg1 composition 1244:1254 the glycan composition 1233:1254 the glycan composition 1233:1254 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 1 15 theme functional 241:250 arg1 roles 252:256 the functional roles 237:256 the functional roles of protein glycosylation 237:281 Glycoproteomics investigates glycan moieties in a site specific manner to reveal the functional roles of protein glycosylation. 29069609 6 16 theme cycle 1013:1017 arg1 limitation 1019:1028 the duty cycle limitation 1004:1028 the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma 1004:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 8 17 theme sites 1408:1412 arg1 detection 1376:1384 detection 1376:1384 detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest 1376:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 11 18 gly glycopeptides 1848:1860 arg2 glycopeptides 1848:1860 glycopeptides 1848:1860 glycopeptides carrying unexpected glycans 1848:1888 The novelty is that this approach does not require glycan composition to be pre-defined, thereby allowing glycopeptides carrying unexpected glycans to be identified. 29069609 7 19 theme unexpected 1319:1328 arg1 structures 1330:1339 unexpected structures 1319:1339 unexpected structures 1319:1339 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 1 20 theme glycan 185:190 arg1 moieties 192:199 glycan moieties 185:199 glycan moieties 185:199 Glycoproteomics investigates glycan moieties in a site specific manner to reveal the functional roles of protein glycosylation. 29069609 6 21 theme samples 1083:1089 arg1 analysis 1063:1070 analysis 1063:1070 analysis of complex samples such as plasma 1063:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 5 22 with case 977:980 arg1 DDA 987:989 DDA 987:989 DDA 987:989 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 8 23 gly N-glycosylation 1392:1406 arg2 sites 1408:1412 59 N-glycosylation sites 1389:1412 59 N-glycosylation sites 1389:1412 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 8 23 gly N-glycosylation 1392:1406 arg2 59 1389:1390 59 1389:1390 59 1389:1390 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 8 24 theme HILIC 1443:1447 arg1 digest 1479:1484 a HILIC enriched human plasma tryptic digest 1441:1484 a HILIC enriched human plasma tryptic digest 1441:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 8 25 theme human 1458:1462 arg1 digest 1479:1484 a HILIC enriched human plasma tryptic digest 1441:1484 a HILIC enriched human plasma tryptic digest 1441:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 0 26 theme predefined 113:122 arg1 knowledge 145:153 predefined glycan compositional knowledge 113:153 predefined glycan compositional knowledge 113:153 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 8 27 gly glycoproteins 1422:1434 arg1 glycoproteins 1422:1434 41 glycoproteins 1419:1434 41 glycoproteins from a HILIC enriched human plasma tryptic digest 1419:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 0 28 gly glycopeptide 83:94 arg2 glycopeptide 83:94 glycopeptide analysis 83:103 glycopeptide analysis 83:103 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 8 29 theme tryptic 1471:1477 arg1 digest 1479:1484 a HILIC enriched human plasma tryptic digest 1441:1484 a HILIC enriched human plasma tryptic digest 1441:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 5 30 theme fetuin 899:904 arg1 analysis 880:887 targeted analysis 871:887 targeted analysis of bovine fetuin 871:904 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 2 31 from Identification 284:297 arg1 acquisition 336:346 data-dependent acquisition 321:346 data-dependent acquisition (DDA) 321:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 2 31 from Identification 284:297 arg1 DDA 349:351 DDA 349:351 DDA 349:351 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 7 32 gly N-glycopeptides 1298:1312 arg2 N-glycopeptides 1298:1312 N-glycopeptides 1298:1312 N-glycopeptides with unexpected structures 1298:1339 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 7 33 theme composition 1244:1254 arg1 knowledge 1220:1228 pre-defined knowledge 1208:1228 pre-defined knowledge of the glycan composition 1208:1254 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 9 34 gly glycoforms 1490:1499 arg1 glycopeptides 1509:1521 IgG1 glycopeptides 1504:1521 IgG1 glycopeptides 1504:1521 21 glycoforms of IgG1 glycopeptides were identified including two truncated structures that are rarely reported. 29069609 12 35 gly glycoforms 2012:2021 arg1 IgG1 2007:2010 two IgG1 glycoforms 2003:2021 two IgG1 glycoforms that are rarely observed 2003:2046 This is demonstrated through the analysis of immunoglobulins in human plasma where we detected two IgG1 glycoforms that are rarely observed. 29069609 0 36 theme independent 22:32 arg1 acquisition 34:44 a data independent acquisition 15:44 a data independent acquisition mass spectrometry 15:62 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 5 37 theme targeted 871:878 arg1 analysis 880:887 targeted analysis 871:887 targeted analysis of bovine fetuin 871:904 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 0 38 theme mass 46:49 arg1 spectrometry 51:62 a data independent acquisition mass spectrometry 15:62 a data independent acquisition mass spectrometry 15:62 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 10 39 theme data-independent 1629:1644 arg1 spectrometry 1651:1662 a data-independent mass spectrometry 1627:1662 a data-independent mass spectrometry 1627:1662 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 2 40 theme quality 369:375 arg1 spectra 383:389 high quality MS/MS spectra 364:389 high quality MS/MS spectra of glycopeptide precursors 364:416 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 12 41 theme IgG1 2007:2010 arg1 glycoforms 2012:2021 two IgG1 glycoforms 2003:2021 two IgG1 glycoforms that are rarely observed 2003:2046 This is demonstrated through the analysis of immunoglobulins in human plasma where we detected two IgG1 glycoforms that are rarely observed. 29069609 4 42 theme data 666:669 arg1 acquisition 671:681 data acquisition 666:681 data acquisition 666:681 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 10 43 theme specific 1685:1692 arg1 glycopeptides 1694:1706 specific glycopeptides 1685:1706 specific glycopeptides from complex biological mixtures 1685:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 11 44 theme unexpected 1871:1880 arg1 glycans 1882:1888 unexpected glycans 1871:1888 unexpected glycans 1871:1888 The novelty is that this approach does not require glycan composition to be pre-defined, thereby allowing glycopeptides carrying unexpected glycans to be identified. 29069609 1 45 theme protein 261:267 arg1 glycosylation 269:281 protein glycosylation 261:281 protein glycosylation 261:281 Glycoproteomics investigates glycan moieties in a site specific manner to reveal the functional roles of protein glycosylation. 29069609 7 46 theme pre-defined 1208:1218 arg1 knowledge 1220:1228 pre-defined knowledge 1208:1228 pre-defined knowledge of the glycan composition 1208:1254 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 11 47 theme glycan 1793:1798 arg1 composition 1800:1810 glycan composition 1793:1810 glycan composition 1793:1810 The novelty is that this approach does not require glycan composition to be pre-defined, thereby allowing glycopeptides carrying unexpected glycans to be identified. 29069609 2 48 theme glycopeptide 394:405 arg1 precursors 407:416 glycopeptide precursors 394:416 glycopeptide precursors 394:416 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 8 49 from digest 1479:1484 arg1 glycoproteins 1422:1434 41 glycoproteins 1419:1434 41 glycoproteins from a HILIC enriched human plasma tryptic digest 1419:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 9 50 theme glycopeptides 1509:1521 arg1 glycoforms 1490:1499 21 glycoforms 1487:1499 21 glycoforms of IgG1 glycopeptides 1487:1521 21 glycoforms of IgG1 glycopeptides were identified including two truncated structures that are rarely reported. 29069609 10 51 theme biological 1721:1730 arg1 mixtures 1732:1739 complex biological mixtures 1713:1739 complex biological mixtures 1713:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 3 52 theme alternative 581:591 arg1 acquisition 560:570 data-independent acquisition 543:570 data-independent acquisition (DIA) 543:576 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 52 theme alternative 581:591 arg1 strategies 605:614 alternative acquisition strategies 581:614 alternative acquisition strategies for glycopeptide analysis 581:640 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 52 theme alternative 581:591 arg1 pseudo-MRM 519:528 pseudo-MRM 519:528 pseudo-MRM (MRM-HR) 519:537 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 7 53 theme DIA 1161:1163 arg1 method 1165:1170 a targeted DIA method 1150:1170 a targeted DIA method 1150:1170 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 2 54 theme data-dependent 321:334 arg1 acquisition 336:346 data-dependent acquisition 321:346 data-dependent acquisition (DDA) 321:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 2 54 theme data-dependent 321:334 arg1 DDA 349:351 DDA 349:351 DDA 349:351 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 6 55 dep trialed 1109:1115 arg1 overcome 995:1002 overcome 995:1002 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma 992:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 3 56 theme glycopeptide 620:631 arg1 analysis 633:640 glycopeptide analysis 620:640 glycopeptide analysis 620:640 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 1 57 theme specific 211:218 arg1 manner 220:225 a site specific manner to reveal the functional roles of protein glycosylation 204:281 a site specific manner to reveal the functional roles of protein glycosylation 204:281 Glycoproteomics investigates glycan moieties in a site specific manner to reveal the functional roles of protein glycosylation. 29069609 4 58 theme re-analysis 728:738 arg1 post-acquisition 740:755 data re-analysis post-acquisition 723:755 data re-analysis post-acquisition 723:755 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 6 59 theme MRM-HR 1033:1038 arg1 acquisition 1040:1050 MRM-HR acquisition 1033:1050 MRM-HR acquisition needed for analysis of complex samples such as plasma 1033:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 4 60 theme full 692:695 arg1 range 708:712 the full MS/MS scan range 688:712 the full MS/MS scan range allowing data re-analysis post-acquisition 688:755 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 3 61 theme data-independent 543:558 arg1 acquisition 560:570 data-independent acquisition 543:570 data-independent acquisition (DIA) 543:576 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 61 theme data-independent 543:558 arg1 pseudo-MRM 519:528 pseudo-MRM 519:528 pseudo-MRM (MRM-HR) 519:537 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 61 theme data-independent 543:558 arg1 strategies 605:614 alternative acquisition strategies 581:614 alternative acquisition strategies for glycopeptide analysis 581:640 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 3 61 theme data-independent 543:558 arg1 DIA 573:575 DIA 573:575 DIA 573:575 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 6 62 theme duty 1008:1011 arg1 limitation 1019:1028 the duty cycle limitation 1004:1028 the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma 1004:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 10 63 gly glycopeptides 1694:1706 arg1 mixtures 1732:1739 complex biological mixtures 1713:1739 complex biological mixtures 1713:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 10 63 gly glycopeptides 1694:1706 arg2 glycopeptides 1694:1706 specific glycopeptides 1685:1706 specific glycopeptides from complex biological mixtures 1685:1739 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 3 64 gly glycopeptide 620:631 arg2 glycopeptide 620:631 glycopeptide analysis 620:640 glycopeptide analysis 620:640 In this study, we investigated pseudo-MRM (MRM-HR) and data-independent acquisition (DIA) as alternative acquisition strategies for glycopeptide analysis. 29069609 2 65 theme glycopeptides 302:314 arg1 Identification 284:297 Identification 284:297 Identification of glycopeptides from data-dependent acquisition (DDA) 284:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 11 66 contain carrying 1862:1869 arg1 glycopeptides 1848:1860 glycopeptides 1848:1860 glycopeptides carrying unexpected glycans 1848:1888 The novelty is that this approach does not require glycan composition to be pre-defined, thereby allowing glycopeptides carrying unexpected glycans to be identified. 29069609 11 66 contain carrying 1862:1869 arg2 glycans 1882:1888 unexpected glycans 1871:1888 unexpected glycans 1871:1888 The novelty is that this approach does not require glycan composition to be pre-defined, thereby allowing glycopeptides carrying unexpected glycans to be identified. 29069609 7 67 with N-glycopeptides 1298:1312 arg1 structures 1330:1339 unexpected structures 1319:1339 unexpected structures 1319:1339 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 8 68 theme N-glycosylation 1392:1406 arg1 sites 1408:1412 59 N-glycosylation sites 1389:1412 59 N-glycosylation sites 1389:1412 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 5 69 theme N-glycopeptide 824:837 arg1 detection 839:847 N-glycopeptide detection 824:847 N-glycopeptide detection 824:847 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 2 70 from acquisition 336:346 arg1 Identification 284:297 Identification 284:297 Identification of glycopeptides from data-dependent acquisition (DDA) 284:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 2 70 from acquisition 336:346 arg1 glycopeptides 302:314 glycopeptides 302:314 glycopeptides from data-dependent acquisition (DDA) 302:352 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 0 71 theme glycopeptide 83:94 arg1 analysis 96:103 glycopeptide analysis 83:103 glycopeptide analysis 83:103 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 6 72 theme complex 1075:1081 arg1 samples 1083:1089 complex samples 1075:1089 complex samples such as plasma 1075:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 6 72 theme complex 1075:1081 arg1 plasma 1099:1104 plasma 1099:1104 plasma 1099:1104 To overcome the duty cycle limitation of MRM-HR acquisition needed for analysis of complex samples such as plasma we trialed DIA. 29069609 4 73 theme data 766:769 arg1 re-acquisition 771:784 data re-acquisition 766:784 data re-acquisition 766:784 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 5 74 theme MRM-HR 804:809 arg1 advantage 791:799 The advantage 787:799 The advantage of MRM-HR over DDA for N-glycopeptide detection 787:847 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 5 75 gly N-glycopeptide 824:837 arg2 N-glycopeptide 824:837 N-glycopeptide detection 824:847 N-glycopeptide detection 824:847 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 0 76 theme glycan 124:129 arg1 knowledge 145:153 predefined glycan compositional knowledge 113:153 predefined glycan compositional knowledge 113:153 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 7 77 gly N-glycopeptides 1184:1198 arg2 N-glycopeptides 1184:1198 N-glycopeptides 1184:1198 N-glycopeptides 1184:1198 This allowed development of a targeted DIA method to identify N-glycopeptides without pre-defined knowledge of the glycan composition, thus providing the potential to identify N-glycopeptides with unexpected structures. 29069609 8 78 theme enriched 1449:1456 arg1 digest 1479:1484 a HILIC enriched human plasma tryptic digest 1441:1484 a HILIC enriched human plasma tryptic digest 1441:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 2 79 theme manual 437:442 arg1 validation 444:453 manual validation 437:453 manual validation to ensure confident assignments 437:485 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 5 80 theme bovine 892:897 arg1 fetuin 899:904 bovine fetuin 892:904 bovine fetuin 892:904 The advantage of MRM-HR over DDA for N-glycopeptide detection was demonstrated from targeted analysis of bovine fetuin where all three N-glycosylation sites were detected, which was not the case with DDA. 29069609 8 81 from glycoproteins 1422:1434 arg1 detection 1376:1384 detection 1376:1384 detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest 1376:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 8 82 theme plasma 1464:1469 arg1 digest 1479:1484 a HILIC enriched human plasma tryptic digest 1441:1484 a HILIC enriched human plasma tryptic digest 1441:1484 This workflow was demonstrated by detection of 59 N-glycosylation sites from 41 glycoproteins from a HILIC enriched human plasma tryptic digest. 29069609 0 83 theme data 17:20 arg1 acquisition 34:44 a data independent acquisition 15:44 a data independent acquisition mass spectrometry 15:62 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 2 84 theme confident 465:473 arg1 assignments 475:485 confident assignments 465:485 confident assignments 465:485 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 0 85 theme acquisition 34:44 arg1 spectrometry 51:62 a data independent acquisition mass spectrometry 15:62 a data independent acquisition mass spectrometry 15:62 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 4 86 dep range 708:712 arg1 scan 703:706 scan 703:706 scan 703:706 These approaches allow data acquisition over the full MS/MS scan range allowing data re-analysis post-acquisition, without data re-acquisition. 29069609 2 87 gly glycopeptide 394:405 arg2 glycopeptide 394:405 glycopeptide 394:405 glycopeptide 394:405 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 0 88 theme spectrometry 51:62 arg1 Development 0:10 Development 0:10 Development of a data independent acquisition mass spectrometry 0:62 Development of a data independent acquisition mass spectrometry workflow to enable glycopeptide analysis without predefined glycan compositional knowledge. 29069609 12 89 theme human 1972:1976 arg1 plasma 1978:1983 human plasma 1972:1983 human plasma where we detected two IgG1 glycoforms that are rarely observed 1972:2046 This is demonstrated through the analysis of immunoglobulins in human plasma where we detected two IgG1 glycoforms that are rarely observed. 29069609 10 90 theme mass 1646:1649 arg1 spectrometry 1651:1662 a data-independent mass spectrometry 1627:1662 a data-independent mass spectrometry 1627:1662 SIGNIFICANCE: We developed a data-independent mass spectrometry workflow to identify specific glycopeptides from complex biological mixtures. 29069609 2 91 theme MS/MS 377:381 arg1 spectra 383:389 high quality MS/MS spectra 364:389 high quality MS/MS spectra of glycopeptide precursors 364:416 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 9 92 gly glycopeptides 1509:1521 arg2 glycopeptides 1509:1521 IgG1 glycopeptides 1504:1521 IgG1 glycopeptides 1504:1521 21 glycoforms of IgG1 glycopeptides were identified including two truncated structures that are rarely reported. 29069609 2 93 theme high 364:367 arg1 spectra 383:389 high quality MS/MS spectra 364:389 high quality MS/MS spectra of glycopeptide precursors 364:416 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 1 94 theme glycosylation 269:281 arg1 roles 252:256 the functional roles 237:256 the functional roles of protein glycosylation 237:281 Glycoproteomics investigates glycan moieties in a site specific manner to reveal the functional roles of protein glycosylation. 29069609 2 95 theme precursors 407:416 arg1 spectra 383:389 high quality MS/MS spectra 364:389 high quality MS/MS spectra of glycopeptide precursors 364:416 Identification of glycopeptides from data-dependent acquisition (DDA) relies on high quality MS/MS spectra of glycopeptide precursors and often requires manual validation to ensure confident assignments. 29069609 9 96 theme IgG1 1504:1507 arg1 glycopeptides 1509:1521 IgG1 glycopeptides 1504:1521 IgG1 glycopeptides 1504:1521 21 glycoforms of IgG1 glycopeptides were identified including two truncated structures that are rarely reported. 26059044 0 0 theme interleukin-22 82:95 arg1 composition 52:62 the atypical N-glycan composition 30:62 the atypical N-glycan composition of plant-produced interleukin-22 30:95 The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity. 26059044 5 1 theme N-glycan 741:748 arg1 analysis 750:757 N-glycan analysis 741:757 N-glycan analysis 741:757 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 8 2 theme Asn54 1095:1099 arg1 mutagenesis 1080:1090 site-directed mutagenesis 1066:1090 site-directed mutagenesis of Asn54 and in vivo deglycosylation 1066:1127 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 0 3 theme plant-produced 67:80 arg1 interleukin-22 82:95 plant-produced interleukin-22 67:95 plant-produced interleukin-22 67:95 The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity. 26059044 0 4 from N-glycan 4:11 arg1 Asn54 16:20 Asn54 16:20 Asn54 16:20 The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity. 26059044 8 5 gly N-glycosylation 1162:1176 arg1 position 1186:1193 this position 1181:1193 this position 1181:1193 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 8 5 gly N-glycosylation 1162:1176 arg2 position 1186:1193 this position 1181:1193 this position 1181:1193 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 8 5 gly N-glycosylation 1162:1176 arg1 protein 1222:1228 proper protein folding 1215:1236 proper protein folding 1215:1236 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 3 6 theme epithelial 482:491 arg1 function 501:508 epithelial barrier function 482:508 epithelial barrier function 482:508 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 4 7 theme human 551:555 arg1 IL-22 557:561 human IL-22 551:561 human IL-22 in Nicotiana benthamiana plants 551:593 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 9 8 from composition 1401:1411 arg1 positions 1435:1443 other positions 1429:1443 other positions 1429:1443 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 10 9 gly glycoproteins 1597:1609 arg1 IL-22 1620:1624 IL-22 1620:1624 IL-22 1620:1624 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 10 9 gly glycoproteins 1597:1609 arg1 glycoproteins 1597:1609 recombinant glycoproteins 1585:1609 recombinant glycoproteins 1585:1609 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 10 10 theme glycoproteins 1597:1609 arg1 activity 1573:1580 activity 1573:1580 activity 1573:1580 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 10 10 theme glycoproteins 1597:1609 arg1 folding 1561:1567 folding 1561:1567 folding 1561:1567 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 4 11 theme Nicotiana 566:574 arg1 plants 588:593 Nicotiana benthamiana plants 566:593 Nicotiana benthamiana plants 566:593 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 6 12 from N-glycans 861:869 arg1 IL-22 874:878 IL-22 874:878 IL-22 874:878 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 6 13 from FUT8 903:906 arg1 plants 918:923 ΔXT/FT plants 911:923 ΔXT/FT plants 911:923 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 4 14 from IL-22 557:561 arg1 plants 588:593 Nicotiana benthamiana plants 566:593 Nicotiana benthamiana plants 566:593 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 9 15 theme atypical 1319:1326 arg1 composition 1337:1347 the atypical N-glycan composition 1315:1347 the atypical N-glycan composition of plant-produced IL-22 1315:1371 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 9 16 theme other 1429:1433 arg1 positions 1435:1443 other positions 1429:1443 other positions 1429:1443 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 4 17 theme protein 643:649 arg1 folding 651:657 protein folding 643:657 protein folding 643:657 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 6 18 from IL-22 874:878 arg1 engineering 835:845 engineering 835:845 engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants 835:923 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 9 19 theme N-glycan 1282:1289 arg1 presence 1268:1275 the presence 1264:1275 the presence of a N-glycan on Asn54 1264:1298 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 9 20 theme IL-22 1367:1371 arg1 composition 1337:1347 the atypical N-glycan composition 1315:1347 the atypical N-glycan composition of plant-produced IL-22 1315:1371 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 9 21 attach presence 1268:1275 arg1 Asn54 1294:1298 Asn54 1294:1298 Asn54 1294:1298 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 9 21 attach presence 1268:1275 arg2 N-glycan 1282:1289 a N-glycan 1280:1289 a N-glycan 1280:1289 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 5 22 theme 5.4 μg/mg 722:730 arg1 TSP 732:734 5.4 μg/mg TSP 722:734 5.4 μg/mg TSP 722:734 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 6 23 from engineering 835:845 arg1 IL-22 874:878 IL-22 874:878 IL-22 874:878 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 6 24 theme N-glycans 861:869 arg1 engineering 835:845 engineering 835:845 engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants 835:923 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 2 25 theme unusual 283:289 arg1 cytokine 291:298 an unusual cytokine 280:298 an unusual cytokine as it does not act directly on immune cells 280:342 IL-22 is an unusual cytokine as it does not act directly on immune cells. 26059044 2 25 theme unusual 283:289 arg1 IL-22 271:275 IL-22 271:275 IL-22 271:275 IL-22 is an unusual cytokine as it does not act directly on immune cells. 26059044 8 26 theme mutagenesis 1080:1090 arg1 combination 1051:1061 The combination 1047:1061 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F 1047:1141 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 4 27 theme N-glycosylation 624:638 arg1 role 616:619 the role 612:619 the role of N-glycosylation on protein folding and biological activity 612:681 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 5 28 theme atypical 788:795 arg1 structure 805:813 the atypical Lewis A structure 784:813 the atypical Lewis A structure 784:813 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 1 29 theme major 242:246 arg1 potential 260:268 major therapeutic potential 242:268 major therapeutic potential 242:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 8 30 theme PNGase 1134:1139 arg1 F 1141:1141 PNGase F 1134:1141 PNGase F 1134:1141 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 2 31 theme immune 331:336 arg1 cells 338:342 immune cells 331:342 immune cells 331:342 IL-22 is an unusual cytokine as it does not act directly on immune cells. 26059044 5 32 theme Lewis 797:801 arg1 structure 805:813 the atypical Lewis A structure 784:813 the atypical Lewis A structure 784:813 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 1 33 theme therapeutic 248:258 arg1 potential 260:268 major therapeutic potential 242:268 major therapeutic potential 242:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 7 34 theme IL-22 1040:1044 arg1 activity 1028:1035 the biological activity 1013:1035 the biological activity of IL-22 1013:1044 Also, core α1,6-fucoylation did not improve the biological activity of IL-22. 26059044 9 35 theme N-glycans 1416:1424 arg1 composition 1401:1411 the N-glycan composition 1388:1411 the N-glycan composition of N-glycans on other positions 1388:1443 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 5 36 theme structure 805:813 arg1 presence 772:779 the presence 768:779 the presence of the atypical Lewis A structure 768:813 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 5 37 theme A 803:803 arg1 structure 805:813 the atypical Lewis A structure 784:813 the atypical Lewis A structure 784:813 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 6 38 from reduction 934:942 arg1 A 953:953 Lewis A 947:953 Lewis A 947:953 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 6 39 theme human-like 850:859 arg1 N-glycans 861:869 human-like N-glycans 850:869 human-like N-glycans on IL-22 850:878 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 4 40 from role 616:619 arg1 folding 651:657 protein folding 643:657 protein folding 643:657 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 4 40 from role 616:619 arg1 activity 674:681 biological activity 663:681 biological activity 663:681 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 1 41 theme Human 135:139 arg1 interleukin-22 141:154 Human interleukin-22 135:154 Human interleukin-22 (IL-22) 135:162 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 1 41 theme Human 135:139 arg1 member 169:174 a member 167:174 a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential 167:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 1 41 theme Human 135:139 arg1 IL-22 157:161 IL-22 157:161 IL-22 157:161 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 10 42 theme N-glycosylation 1542:1556 arg1 role 1534:1537 the role 1530:1537 the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22 1530:1624 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 5 43 theme IL-22 705:709 arg1 levels 695:700 Expression levels 684:700 Expression levels of IL-22 684:709 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 1 44 theme IL-10 183:187 arg1 family 198:203 the IL-10 cytokine family 179:203 the IL-10 cytokine family 179:203 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 3 45 dep differentiation 373:387 arg1 the 369:371 the 369:371 the 369:371 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 3 46 theme antimicrobial 408:420 arg1 expression 430:439 antimicrobial protein expression 408:439 antimicrobial protein expression 408:439 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 8 47 theme protein 1222:1228 arg1 folding 1230:1236 proper protein folding 1215:1236 proper protein folding 1215:1236 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 6 48 theme strong 927:932 arg1 reduction 934:942 a strong reduction 925:942 a strong reduction in Lewis A 925:953 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 9 49 theme N-glycan 1392:1399 arg1 composition 1401:1411 the N-glycan composition 1388:1411 the N-glycan composition of N-glycans on other positions 1388:1443 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 10 50 from role 1534:1537 arg1 activity 1573:1580 activity 1573:1580 activity 1573:1580 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 10 50 from role 1534:1537 arg1 folding 1561:1567 folding 1561:1567 folding 1561:1567 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 1 51 theme cytokine 189:196 arg1 family 198:203 the IL-10 cytokine family 179:203 the IL-10 cytokine family 179:203 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 0 52 theme N-glycan 43:50 arg1 composition 52:62 the atypical N-glycan composition 30:62 the atypical N-glycan composition of plant-produced interleukin-22 30:95 The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity. 26059044 9 53 theme N-glycan 1328:1335 arg1 composition 1337:1347 the atypical N-glycan composition 1315:1347 the atypical N-glycan composition of plant-produced IL-22 1315:1371 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 6 54 theme Lewis 947:951 arg1 A 953:953 Lewis A 947:953 Lewis A 947:953 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 10 55 theme excellent 1500:1508 arg1 tool 1510:1513 an excellent tool 1497:1513 an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22 1497:1624 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 0 56 theme atypical 34:41 arg1 composition 52:62 the atypical N-glycan composition 30:62 the atypical N-glycan composition of plant-produced interleukin-22 30:95 The N-glycan on Asn54 affects the atypical N-glycan composition of plant-produced interleukin-22, but does not influence its activity. 26059044 7 57 theme biological 1017:1026 arg1 activity 1028:1035 the biological activity 1013:1035 the biological activity of IL-22 1013:1044 Also, core α1,6-fucoylation did not improve the biological activity of IL-22. 26059044 6 58 theme mouse 897:901 arg1 FUT8 903:906 mouse FUT8 897:906 mouse FUT8 in ΔXT/FT plants 897:923 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 6 59 theme ΔXT/FT 911:916 arg1 plants 918:923 ΔXT/FT plants 911:923 ΔXT/FT plants 911:923 Surprisingly, upon engineering of human-like N-glycans on IL-22 by co-expressing mouse FUT8 in ΔXT/FT plants a strong reduction in Lewis A was observed. 26059044 8 60 theme site-directed 1066:1078 arg1 mutagenesis 1080:1090 site-directed mutagenesis 1066:1090 site-directed mutagenesis of Asn54 and in vivo deglycosylation 1066:1127 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 1 61 contain have 237:240 arg1 interleukin-22 141:154 Human interleukin-22 135:154 Human interleukin-22 (IL-22) 135:162 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 1 61 contain have 237:240 arg1 member 169:174 a member 167:174 a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential 167:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 1 61 contain have 237:240 arg2 potential 260:268 major therapeutic potential 242:268 major therapeutic potential 242:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 7 62 theme core 975:978 arg1 α1,6-fucoylation 980:995 core α1,6-fucoylation 975:995 core α1,6-fucoylation 975:995 Also, core α1,6-fucoylation did not improve the biological activity of IL-22. 26059044 3 63 theme epithelial 444:453 arg1 cells 455:459 epithelial cells 444:459 epithelial cells 444:459 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 9 64 theme plant-produced 1352:1365 arg1 IL-22 1367:1371 plant-produced IL-22 1352:1371 plant-produced IL-22 1352:1371 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 1 65 theme family 198:203 arg1 interleukin-22 141:154 Human interleukin-22 135:154 Human interleukin-22 (IL-22) 135:162 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 1 65 theme family 198:203 arg1 member 169:174 a member 167:174 a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential 167:268 Human interleukin-22 (IL-22) is a member of the IL-10 cytokine family that has recently been shown to have major therapeutic potential. 26059044 5 66 theme Expression 684:693 arg1 levels 695:700 Expression levels 684:700 Expression levels of IL-22 684:709 Expression levels of IL-22 were up to 5.4 μg/mg TSP, and N-glycan analysis revealed the presence of the atypical Lewis A structure. 26059044 3 67 theme cells 455:459 arg1 proliferation 390:402 proliferation 390:402 proliferation 390:402 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 3 67 theme cells 455:459 arg1 differentiation 373:387 differentiation 373:387 differentiation 373:387 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 3 67 theme cells 455:459 arg1 expression 430:439 antimicrobial protein expression 408:439 antimicrobial protein expression 408:439 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 8 68 with combination 1051:1061 arg1 F 1141:1141 PNGase F 1134:1141 PNGase F 1134:1141 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 4 69 theme benthamiana 576:586 arg1 plants 588:593 Nicotiana benthamiana plants 566:593 Nicotiana benthamiana plants 566:593 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26059044 8 70 theme deglycosylation 1113:1127 arg1 mutagenesis 1080:1090 site-directed mutagenesis 1066:1090 site-directed mutagenesis of Asn54 and in vivo deglycosylation 1066:1127 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 3 71 theme protein 422:428 arg1 expression 430:439 antimicrobial protein expression 408:439 antimicrobial protein expression 408:439 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 3 72 theme barrier 493:499 arg1 function 501:508 epithelial barrier function 482:508 epithelial barrier function 482:508 Instead, IL-22 controls the differentiation, proliferation and antimicrobial protein expression of epithelial cells, thereby maintaining epithelial barrier function. 26059044 8 73 theme proper 1215:1220 arg1 folding 1230:1236 proper protein folding 1215:1236 proper protein folding 1215:1236 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 9 74 from presence 1268:1275 arg1 Asn54 1294:1298 Asn54 1294:1298 Asn54 1294:1298 However, we do show that the presence of a N-glycan on Asn54 contributes to the atypical N-glycan composition of plant-produced IL-22 and influences the N-glycan composition of N-glycans on other positions. 26059044 8 75 theme in vivo 1105:1111 arg1 deglycosylation 1113:1127 in vivo deglycosylation 1105:1127 in vivo deglycosylation 1105:1127 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 8 76 from position 1186:1193 arg1 N-glycosylation 1162:1176 N-glycosylation 1162:1176 N-glycosylation at this position 1162:1193 The combination of site-directed mutagenesis of Asn54 and in vivo deglycosylation with PNGase F also revealed that N-glycosylation at this position is not required for proper protein folding. 26059044 10 77 theme recombinant 1585:1595 arg1 IL-22 1620:1624 IL-22 1620:1624 IL-22 1620:1624 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 10 77 theme recombinant 1585:1595 arg1 glycoproteins 1597:1609 recombinant glycoproteins 1585:1609 recombinant glycoproteins 1585:1609 Altogether, our data demonstrate that plants offer an excellent tool to investigate the role of N-glycosylation on folding and activity of recombinant glycoproteins, such as IL-22. 26059044 4 78 theme biological 663:672 arg1 activity 674:681 biological activity 663:681 biological activity 663:681 In this study, we transiently expressed human IL-22 in Nicotiana benthamiana plants and investigated the role of N-glycosylation on protein folding and biological activity. 26984886 6 0 theme combined 842:849 arg1 analysis 851:858 combined analysis 842:858 combined analysis of glycan micro and macro-heterogeneity and bioassays 842:912 Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. 26984886 3 1 theme virulence 531:539 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 1 theme virulence 531:539 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 1 theme virulence 531:539 arg1 determinant 502:512 the primary determinant 490:512 the primary determinant of antigenicity, virulence and zoonotic potential 490:562 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 1 2 theme major 264:268 arg1 burden 281:286 a major healthcare burden 262:286 a major healthcare burden 262:286 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 8 3 theme lectin 1208:1213 arg1 data 1223:1226 immune lectin binding data 1201:1226 immune lectin binding data 1201:1226 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 4 4 theme glycosylation 614:626 arg1 sites 628:632 HA glycosylation sites 611:632 HA glycosylation sites 611:632 Mutations leading to changes in the number of HA glycosylation sites are often reported. 26984886 5 5 theme phenotypic 795:804 arg1 changes 806:812 actual phenotypic changes 788:812 actual phenotypic changes in HA structure 788:828 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 11 6 theme glycan 1771:1776 arg1 enzymes 1789:1795 glycan remodeling enzymes 1771:1795 glycan remodeling enzymes 1771:1795 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 5 7 theme Such 654:657 arg1 studies 678:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 11 8 theme immature 1673:1680 arg1 glycans 1701:1707 immature, high-mannose type glycans 1673:1707 glycans 1701:1707 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 1 9 theme A 199:199 arg1 virus 201:205 influenza A virus 189:205 influenza A virus 189:205 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 9 10 from evolution 1386:1394 arg1 response 1399:1406 response 1399:1406 response to selection pressures and interactions with innate immune-lectins 1399:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 7 11 theme system 1133:1138 arg1 pressure 1140:1147 innate immune system pressure 1119:1147 innate immune system pressure 1119:1147 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 10 12 theme structural 1494:1503 arg1 basis 1505:1509 the structural basis 1490:1509 the structural basis for site-specific glycan microheterogeneity at these sites 1490:1568 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 8 13 theme phenotypic 1245:1254 arg1 changes 1256:1262 the phenotypic changes 1241:1262 the phenotypic changes 1241:1262 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 12 14 theme immune 1855:1860 arg1 recognition 1869:1879 immune lectin recognition 1855:1879 immune lectin recognition 1855:1879 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 7 15 theme innate 1119:1124 arg1 system 1133:1138 innate immune system 1119:1138 innate immune system pressure 1119:1147 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 2 16 theme developing 306:315 arg1 vaccines 317:324 developing vaccines 306:324 developing vaccines against influenza 306:342 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 1 17 theme imminent 218:225 arg1 threat 227:232 an imminent threat 215:232 an imminent threat to the world population and a major healthcare burden 215:286 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 12 18 from sites 1835:1839 arg1 glycans 1824:1830 the high mannose glycans 1807:1830 the high mannose glycans at sites implicated in immune lectin recognition 1807:1879 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 12 18 from sites 1835:1839 arg1 capable 1902:1908 capable 1902:1908 capable 1902:1908 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 5 19 theme N-linked 744:751 arg1 glycosylation 753:765 N-linked glycosylation 744:765 N-linked glycosylation 744:765 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 4 20 from changes 586:592 arg1 number 601:606 the number 597:606 the number of HA glycosylation sites 597:632 Mutations leading to changes in the number of HA glycosylation sites are often reported. 26984886 6 21 theme viral 972:976 arg1 bioactivity 978:988 viral bioactivity 972:988 viral bioactivity 972:988 Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. 26984886 7 22 theme HA 1085:1086 arg1 glycomics 1052:1060 glycomics 1052:1060 glycomics 1052:1060 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 7 22 theme HA 1085:1086 arg1 proteomics 1040:1049 proteomics 1040:1049 proteomics 1040:1049 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 7 22 theme HA 1085:1086 arg1 glycoproteomics 1066:1080 glycoproteomics 1066:1080 glycoproteomics of HA before and after adaptation to innate immune system pressure 1066:1147 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 9 23 theme glycosylation 1336:1348 arg1 sites 1350:1354 the glycosylation sites 1332:1354 the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins 1332:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 1 24 theme sustained 151:159 arg1 effort 181:186 sustained biomedical research effort 151:186 sustained biomedical research effort 151:186 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 1 25 theme research 172:179 arg1 effort 181:186 sustained biomedical research effort 151:186 sustained biomedical research effort 151:186 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 0 26 theme A 86:86 arg1 Virus 88:92 Influenza A Virus Glycan Microheterogeneity and Host Interactions 76:140 Virus 88:92 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 5 27 from changes 806:812 arg1 structure 820:828 HA structure 817:828 HA structure 817:828 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 12 28 theme high 1811:1814 arg1 glycans 1824:1830 the high mannose glycans 1807:1830 the high mannose glycans at sites implicated in immune lectin recognition 1807:1879 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 12 28 theme high 1811:1814 arg1 capable 1902:1908 capable 1902:1908 capable 1902:1908 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 10 29 theme glycan 1529:1534 arg1 microheterogeneity 1536:1553 site-specific glycan microheterogeneity 1515:1553 site-specific glycan microheterogeneity at these sites 1515:1568 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 0 30 theme Glycan 94:99 arg1 Microheterogeneity 101:118 Glycan Microheterogeneity 94:118 Glycan Microheterogeneity 94:118 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 0 31 theme Integrated 0:9 arg1 Omics 11:15 Integrated Omics 0:15 Integrated Omics 0:15 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 9 32 theme selection 1411:1419 arg1 pressures 1421:1429 selection pressures 1411:1429 selection pressures 1411:1429 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 11 33 attach presence 1661:1668 arg1 site 1725:1728 a particular site 1712:1728 a particular site 1712:1728 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 11 33 attach presence 1661:1668 arg2 glycans 1701:1707 immature, high-mannose type glycans 1673:1707 glycans 1701:1707 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 10 34 theme structural 1584:1593 arg1 modeling 1595:1602 structural modeling 1584:1602 structural modeling 1584:1602 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 2 35 theme immune 415:420 arg1 pressure 422:429 selective immune pressure 405:429 selective immune pressure 405:429 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 3 36 theme surface 465:471 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 36 theme surface 465:471 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 6 37 theme macro-heterogeneity 880:898 arg1 analysis 851:858 combined analysis 842:858 combined analysis of glycan micro and macro-heterogeneity and bioassays 842:912 Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. 26984886 3 38 gly glycoprotein 473:484 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 38 gly glycoprotein 473:484 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 0 39 theme Structural 55:64 arg1 Basis 66:70 Structural Basis 55:70 Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions 55:140 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 12 40 theme immune-lectin 1952:1964 arg1 protein-D 1977:1985 the immune-lectin surfactant protein-D 1948:1985 the immune-lectin surfactant protein-D 1948:1985 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 6 41 theme micro 870:874 arg1 analysis 851:858 combined analysis 842:858 combined analysis of glycan micro and macro-heterogeneity and bioassays 842:912 Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. 26984886 10 42 theme dynamics 1618:1625 arg1 simulations 1627:1637 molecular dynamics simulations 1608:1637 molecular dynamics simulations 1608:1637 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 11 43 theme type 1696:1699 arg1 glycans 1701:1707 immature, high-mannose type glycans 1673:1707 glycans 1701:1707 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 3 44 theme primary 494:500 arg1 determinant 502:512 the primary determinant 490:512 the primary determinant of antigenicity, virulence and zoonotic potential 490:562 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 4 45 theme sites 628:632 arg1 number 601:606 the number 597:606 the number of HA glycosylation sites 597:632 Mutations leading to changes in the number of HA glycosylation sites are often reported. 26984886 11 46 theme particular 1714:1723 arg1 site 1725:1728 a particular site 1712:1728 a particular site 1712:1728 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 2 47 theme virus 366:370 arg1 ability 351:357 the ability 347:357 the ability of the virus to mutate rapidly in response to selective immune pressure 347:429 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 2 47 theme virus 366:370 arg1 challenge 293:301 The challenge 289:301 The challenge in developing vaccines against influenza 289:342 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 5 48 theme genetic 659:665 arg1 studies 678:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 11 49 theme reduced 1746:1752 arg1 accessibility 1754:1766 reduced accessibility 1746:1766 reduced accessibility to glycan remodeling enzymes 1746:1795 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 1 50 theme healthcare 270:279 arg1 burden 281:286 a major healthcare burden 262:286 a major healthcare burden 262:286 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 8 51 theme immune 1201:1206 arg1 data 1223:1226 immune lectin binding data 1201:1226 immune lectin binding data 1201:1226 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 4 52 theme HA 611:612 arg1 sites 628:632 HA glycosylation sites 611:632 HA glycosylation sites 611:632 Mutations leading to changes in the number of HA glycosylation sites are often reported. 26984886 9 53 with interactions 1435:1446 arg1 immune-lectins 1460:1473 innate immune-lectins 1453:1473 innate immune-lectins 1453:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 3 54 theme potential 554:562 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 54 theme potential 554:562 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 54 theme potential 554:562 arg1 determinant 502:512 the primary determinant 490:512 the primary determinant of antigenicity, virulence and zoonotic potential 490:562 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 1 55 theme influenza 189:197 arg1 virus 201:205 influenza A virus 189:205 influenza A virus 189:205 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 8 56 theme binding 1215:1221 arg1 data 1223:1226 immune lectin binding data 1201:1226 immune lectin binding data 1201:1226 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 11 57 theme remodeling 1778:1787 arg1 enzymes 1789:1795 glycan remodeling enzymes 1771:1795 glycan remodeling enzymes 1771:1795 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 7 58 theme immune 1126:1131 arg1 system 1133:1138 innate immune system 1119:1138 innate immune system pressure 1119:1147 We present a study that integrates proteomics, glycomics and glycoproteomics of HA before and after adaptation to innate immune system pressure. 26984886 9 59 with pressures 1421:1429 arg1 immune-lectins 1460:1473 innate immune-lectins 1453:1473 innate immune-lectins 1453:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 11 60 from site 1725:1728 arg1 presence 1661:1668 the presence 1657:1668 the presence of immature, high-mannose type glycans at a particular site 1657:1728 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 5 61 theme sequencing 667:676 arg1 studies 678:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies 654:684 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 12 62 theme lectin 1862:1867 arg1 recognition 1869:1879 immune lectin recognition 1855:1879 immune lectin recognition 1855:1879 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 9 63 theme Underprocessed 1290:1303 arg1 glycoforms 1305:1314 Underprocessed glycoforms 1290:1314 Underprocessed glycoforms 1290:1314 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 10 64 from sites 1564:1568 arg1 microheterogeneity 1536:1553 site-specific glycan microheterogeneity 1515:1553 site-specific glycan microheterogeneity at these sites 1515:1568 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 9 65 gly glycosylation 1336:1348 arg2 sites 1350:1354 the glycosylation sites 1332:1354 the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins 1332:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 8 66 theme biological 1269:1278 arg1 activity 1280:1287 biological activity 1269:1287 biological activity 1269:1287 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 12 67 theme mannose 1816:1822 arg1 glycans 1824:1830 the high mannose glycans 1807:1830 the high mannose glycans at sites implicated in immune lectin recognition 1807:1879 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 12 67 theme mannose 1816:1822 arg1 capable 1902:1908 capable 1902:1908 capable 1902:1908 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 0 68 theme Influenza 76:84 arg1 Virus 88:92 Influenza A Virus Glycan Microheterogeneity and Host Interactions 76:140 Virus 88:92 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 1 69 theme biomedical 161:170 arg1 effort 181:186 sustained biomedical research effort 151:186 sustained biomedical research effort 151:186 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 2 70 from challenge 293:301 arg1 vaccines 317:324 developing vaccines 306:324 developing vaccines against influenza 306:342 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 5 71 theme actual 788:793 arg1 changes 806:812 actual phenotypic changes 788:812 actual phenotypic changes in HA structure 788:828 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 5 72 theme sequons 732:738 arg1 creation 720:727 creation 720:727 creation 720:727 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 5 72 theme sequons 732:738 arg1 disruption 706:715 disruption 706:715 disruption 706:715 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 10 73 theme site-specific 1515:1527 arg1 microheterogeneity 1536:1553 site-specific glycan microheterogeneity 1515:1553 site-specific glycan microheterogeneity at these sites 1515:1568 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 0 74 theme Host 124:127 arg1 Interactions 129:140 Host Interactions 124:140 Host Interactions 124:140 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 9 75 theme viral 1380:1384 arg1 evolution 1386:1394 viral evolution 1380:1394 viral evolution in response to selection pressures and interactions with innate immune-lectins 1380:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 5 76 dep disruption 706:715 arg1 the 702:704 the 702:704 the 702:704 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 3 77 theme predominant 453:463 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 77 theme predominant 453:463 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 0 78 dep Virus 88:92 arg1 Microheterogeneity 101:118 Glycan Microheterogeneity 94:118 Glycan Microheterogeneity 94:118 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 0 79 theme Computational 21:33 arg1 Glycobiology 35:46 Computational Glycobiology 21:46 Computational Glycobiology 21:46 Integrated Omics and Computational Glycobiology Reveal Structural Basis for Influenza A Virus Glycan Microheterogeneity and Host Interactions. 26984886 4 80 gly glycosylation 614:626 arg2 sites 628:632 HA glycosylation sites 611:632 HA glycosylation sites 611:632 Mutations leading to changes in the number of HA glycosylation sites are often reported. 26984886 11 81 theme glycans 1701:1707 arg1 presence 1661:1668 the presence 1657:1668 the presence of immature, high-mannose type glycans at a particular site 1657:1728 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 2 82 theme selective 405:413 arg1 pressure 422:429 selective immune pressure 405:429 selective immune pressure 405:429 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 5 83 theme HA 817:818 arg1 structure 820:828 HA structure 817:828 HA structure 817:828 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 26984886 12 84 theme surfactant 1966:1975 arg1 protein-D 1977:1985 the immune-lectin surfactant protein-D 1948:1985 the immune-lectin surfactant protein-D 1948:1985 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 12 85 theme trimeric 1921:1928 arg1 interactions 1930:1941 trimeric interactions 1921:1941 trimeric interactions 1921:1941 Further, the high mannose glycans at sites implicated in immune lectin recognition were predicted to be capable of forming trimeric interactions with the immune-lectin surfactant protein-D. 26984886 2 86 dep pressure 422:429 arg1 response 393:400 response 393:400 response 393:400 The challenge in developing vaccines against influenza is the ability of the virus to mutate rapidly in response to selective immune pressure. 26984886 11 87 theme high-mannose 1683:1694 arg1 glycans 1701:1707 immature, high-mannose type glycans 1673:1707 glycans 1701:1707 We observed that the presence of immature, high-mannose type glycans at a particular site correlated with reduced accessibility to glycan remodeling enzymes. 26984886 3 88 theme antigenicity 517:528 arg1 glycoprotein 473:484 the predominant surface glycoprotein 449:484 the predominant surface glycoprotein 449:484 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 88 theme antigenicity 517:528 arg1 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin 432:444 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 3 88 theme antigenicity 517:528 arg1 determinant 502:512 the primary determinant 490:512 the primary determinant of antigenicity, virulence and zoonotic potential 490:562 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 9 89 theme innate 1453:1458 arg1 immune-lectins 1460:1473 innate immune-lectins 1453:1473 innate immune-lectins 1453:1473 Underprocessed glycoforms predominated at the glycosylation sites found to be involved in viral evolution in response to selection pressures and interactions with innate immune-lectins. 26984886 6 90 theme glycan 863:868 arg1 micro 870:874 glycan micro 863:874 glycan micro 863:874 Therefore, combined analysis of glycan micro and macro-heterogeneity and bioassays will better define the relationships among glycosylation, viral bioactivity and evolution. 26984886 10 91 theme molecular 1608:1616 arg1 simulations 1627:1637 molecular dynamics simulations 1608:1637 molecular dynamics simulations 1608:1637 To understand the structural basis for site-specific glycan microheterogeneity at these sites, we performed structural modeling and molecular dynamics simulations. 26984886 3 92 theme zoonotic 545:552 arg1 potential 554:562 zoonotic potential 545:562 zoonotic potential 545:562 Hemagglutinin is the predominant surface glycoprotein and the primary determinant of antigenicity, virulence and zoonotic potential. 26984886 1 93 theme world 241:245 arg1 population 247:256 the world population 237:256 the world population 237:256 Despite sustained biomedical research effort, influenza A virus remains an imminent threat to the world population and a major healthcare burden. 26984886 8 94 theme glycan 1184:1189 arg1 array 1191:1195 glycan array 1184:1195 glycan array 1184:1195 We combined this information with glycan array and immune lectin binding data to correlate the phenotypic changes with biological activity. 26984886 5 95 link N-linked 744:751 arg1 glycosylation 753:765 N-linked glycosylation 744:765 N-linked glycosylation 744:765 Such genetic sequencing studies predict at best the disruption or creation of sequons for N-linked glycosylation; they do not reflect actual phenotypic changes in HA structure. 25110774 9 0 theme mouse 1695:1699 arg1 liver 1701:1705 mouse liver 1695:1705 mouse liver 1695:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 6 1 from enrichment 980:989 arg1 samples 1036:1042 the complex biological samples 1013:1042 the complex biological samples 1013:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 10 2 theme enriching 1941:1949 arg1 glycopeptides 1964:1976 identifying and enriching low-abundant glycopeptides 1925:1976 identifying and enriching low-abundant glycopeptides 1925:1976 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 10 3 from potential 1911:1919 arg1 samples 2000:2006 complex biological samples 1981:2006 complex biological samples 1981:2006 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 1 4 from samples 178:184 arg1 enrichment 122:131 Selective enrichment 112:131 Selective enrichment of glycopeptides from complicated biological samples 112:184 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 1 5 theme biological 167:176 arg1 samples 178:184 complicated biological samples 155:184 complicated biological samples 155:184 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 6 6 gly glycopeptides 994:1006 arg2 glycopeptides 994:1006 glycopeptides 994:1006 glycopeptides from the complex biological samples 994:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 6 gly glycopeptides 994:1006 arg1 samples 1036:1042 the complex biological samples 1013:1042 the complex biological samples 1013:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 10 7 theme complex 1981:1987 arg1 samples 2000:2006 complex biological samples 1981:2006 complex biological samples 1981:2006 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 3 8 theme methods 591:597 arg1 application 570:580 the application 566:580 the application of these methods 566:597 However, the application of these methods is limited by the medium selectivity of HILIC matrices. 25110774 9 9 theme N-glycosylation 1723:1737 arg1 sites 1739:1743 344 unique N-glycosylation sites 1712:1743 344 unique N-glycosylation sites that mapped to 290 different glycoproteins 1712:1786 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 8 10 with limit 1484:1488 arg1 yield 1530:1534 a satisfactory recovery yield 1506:1534 a satisfactory recovery yield (84-103%) 1506:1544 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 10 with limit 1484:1488 arg1 % 1543:1543 84-103% 1537:1543 84-103% 1537:1543 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 9 11 gly glycoproteins 1774:1786 arg1 glycoproteins 1774:1786 290 different glycoproteins 1760:1786 290 different glycoproteins 1760:1786 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 4 12 theme metal-organic 682:694 arg1 frameworks 696:705 hydrophilic metal-organic frameworks 670:705 hydrophilic metal-organic frameworks (MOFs) 670:712 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 12 theme metal-organic 682:694 arg1 matrix 750:755 a HILIC matrix 742:755 a HILIC matrix 742:755 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 12 theme metal-organic 682:694 arg1 MOFs 708:711 MOFs 708:711 MOFs 708:711 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 1 13 theme glycosylation 246:258 arg1 information 260:270 glycosylation information 246:270 glycosylation information 246:270 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 6 14 theme biological 1025:1034 arg1 samples 1036:1042 the complex biological samples 1013:1042 the complex biological samples 1013:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 5 15 theme nano-sized 861:870 arg1 structure 878:886 nano-sized cubic structure 861:886 nano-sized cubic structure 861:886 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 9 16 from liver 1701:1705 arg1 glycopeptides 1676:1688 glycopeptides 1676:1688 glycopeptides from mouse liver 1676:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 9 16 from liver 1701:1705 arg1 enrichment 1662:1671 the selective enrichment 1648:1671 the selective enrichment of glycopeptides from mouse liver 1648:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 9 17 theme different 1764:1772 arg1 glycoproteins 1774:1786 290 different glycoproteins 1760:1786 290 different glycoproteins 1760:1786 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 8 18 theme satisfactory 1508:1519 arg1 yield 1530:1534 a satisfactory recovery yield 1506:1534 a satisfactory recovery yield (84-103%) 1506:1544 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 18 theme satisfactory 1508:1519 arg1 % 1543:1543 84-103% 1537:1543 84-103% 1537:1543 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 3 19 theme HILIC 639:643 arg1 matrices 645:652 HILIC matrices 639:652 HILIC matrices 639:652 However, the application of these methods is limited by the medium selectivity of HILIC matrices. 25110774 1 20 theme complicated 155:165 arg1 samples 178:184 complicated biological samples 155:184 complicated biological samples 155:184 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 6 21 theme glycopeptides 994:1006 arg1 enrichment 980:989 the selective enrichment 966:989 the selective enrichment of glycopeptides from the complex biological samples 966:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 22 theme immunoglobulin 1095:1108 arg1 samples 1144:1150 standard samples 1135:1150 standard samples 1135:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 22 theme immunoglobulin 1095:1108 arg1 IgG 1113:1115 IgG 1113:1115 IgG 1113:1115 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 22 theme immunoglobulin 1095:1108 arg1 G 1110:1110 human immunoglobulin G 1089:1110 human immunoglobulin G (IgG) that was used as standard samples 1089:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 0 23 theme hydrophilic 72:82 arg1 chromatography 96:109 hydrophilic interaction chromatography 72:109 hydrophilic interaction chromatography 72:109 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 10 24 gly glycopeptides 1964:1976 arg2 glycopeptides 1964:1976 identifying and enriching low-abundant glycopeptides 1925:1976 identifying and enriching low-abundant glycopeptides 1925:1976 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 9 25 theme selective 1652:1660 arg1 enrichment 1662:1671 the selective enrichment 1648:1671 the selective enrichment of glycopeptides from mouse liver 1648:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 2 26 theme matrices 547:554 arg1 glycan 515:520 the glycan 511:520 the glycan of glycopeptides and the matrices 511:554 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 2 27 gly glycopeptides 468:480 arg2 glycopeptides 468:480 glycopeptides 468:480 glycopeptides 468:480 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 6 28 theme digested 1069:1076 arg1 mixture 1078:1084 a digested mixture 1067:1084 a digested mixture of human immunoglobulin G (IgG) that was used as standard samples 1067:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 8 29 theme glycopeptide 1550:1561 arg1 enrichment 1563:1572 glycopeptide enrichment 1550:1572 glycopeptide enrichment from a digest of IgG 1550:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 2 30 theme selective 445:453 arg1 isolation 455:463 selective isolation 445:463 selective isolation of glycopeptides 445:480 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 0 31 theme metal-organic 44:56 arg1 frameworks 58:67 metal-organic frameworks 44:67 metal-organic frameworks 44:67 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 9 32 from enrichment 1662:1671 arg1 liver 1701:1705 mouse liver 1695:1705 mouse liver 1695:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 7 33 theme digest 1407:1412 arg1 fmol 1395:1398 67 fmol 1392:1398 67 fmol of IgG digest 1392:1412 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 0 34 theme interaction 84:94 arg1 chromatography 96:109 hydrophilic interaction chromatography 72:109 hydrophilic interaction chromatography 72:109 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 8 35 theme resulting 1432:1440 arg1 LCD-MOFs 1442:1449 the resulting LCD-MOFs 1428:1449 the resulting LCD-MOFs 1428:1449 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 0 36 theme Efficient 0:8 arg1 enrichment 10:19 Efficient enrichment 0:19 Efficient enrichment of glycopeptides 0:36 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 9 37 theme promising 1611:1619 arg1 protocol 1621:1628 a promising protocol 1609:1628 a promising protocol 1609:1628 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 8 38 theme IgG 1591:1593 arg1 digest 1581:1586 a digest 1579:1586 a digest of IgG 1579:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 6 39 from samples 1036:1042 arg1 glycopeptides 994:1006 glycopeptides 994:1006 glycopeptides from the complex biological samples 994:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 39 from samples 1036:1042 arg1 enrichment 980:989 the selective enrichment 966:989 the selective enrichment of glycopeptides from the complex biological samples 966:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 5 40 theme cubic 872:876 arg1 structure 878:886 nano-sized cubic structure 861:886 nano-sized cubic structure 861:886 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 1 41 theme glycopeptides 136:148 arg1 enrichment 122:131 Selective enrichment 112:131 Selective enrichment of glycopeptides from complicated biological samples 112:184 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 8 42 theme lower 1468:1472 arg1 limit 1484:1488 the lower detection limit 1464:1488 the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG 1464:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 42 theme lower 1468:1472 arg1 fmol 1495:1498 3.3 fmol 1491:1498 3.3 fmol 1491:1498 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 43 from digest 1581:1586 arg1 enrichment 1563:1572 glycopeptide enrichment 1550:1572 glycopeptide enrichment from a digest of IgG 1550:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 1 44 theme great 341:345 arg1 challenge 347:355 a great challenge 339:355 a great challenge 339:355 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 0 45 gly glycopeptides 24:36 arg2 glycopeptides 24:36 glycopeptides 24:36 glycopeptides 24:36 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 2 46 theme chromatography 382:395 arg1 strategies 411:420 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 1 47 theme glycoproteins 275:287 arg1 structure 232:240 structure 232:240 structure 232:240 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 1 47 theme glycoproteins 275:287 arg1 information 260:270 glycosylation information 246:270 glycosylation information 246:270 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 10 48 theme great 1905:1909 arg1 potential 1911:1919 great potential 1905:1919 great potential for identifying and enriching low-abundant glycopeptides in complex biological samples 1905:2006 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 1 49 theme mass 295:298 arg1 MS 314:315 MS 314:315 MS 314:315 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 1 49 theme mass 295:298 arg1 spectrometry 300:311 mass spectrometry 295:311 mass spectrometry (MS) 295:316 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 9 50 theme single 1809:1814 arg1 run 1819:1821 a single MS run 1807:1821 a single MS run 1807:1821 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 2 51 theme -based 404:409 arg1 strategies 411:420 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 6 52 theme selective 970:978 arg1 enrichment 980:989 the selective enrichment 966:989 the selective enrichment of glycopeptides from the complex biological samples 966:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 7 53 theme selectivity 1160:1170 arg1 assessment 1172:1181 the selectivity assessment 1156:1181 the selectivity assessment 1156:1181 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 9 54 gly N-glycosylation 1723:1737 arg2 sites 1739:1743 344 unique N-glycosylation sites 1712:1743 344 unique N-glycosylation sites that mapped to 290 different glycoproteins 1712:1786 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 9 54 gly N-glycosylation 1723:1737 arg2 344 1712:1714 344 1712:1714 344 1712:1714 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 3 55 theme medium 617:622 arg1 selectivity 624:634 the medium selectivity 613:634 the medium selectivity of HILIC matrices 613:652 However, the application of these methods is limited by the medium selectivity of HILIC matrices. 25110774 9 56 gly glycopeptides 1676:1688 arg2 glycopeptides 1676:1688 glycopeptides 1676:1688 glycopeptides from mouse liver 1676:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 9 56 gly glycopeptides 1676:1688 arg1 liver 1701:1705 mouse liver 1695:1705 mouse liver 1695:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 10 57 theme biological 1989:1998 arg1 samples 2000:2006 complex biological samples 1981:2006 complex biological samples 1981:2006 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 5 58 link cross-linked 762:773 arg1 LCD-MOFs 784:791 LCD-MOFs 784:791 LCD-MOFs 784:791 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 5 58 link cross-linked 762:773 arg1 CD-MOFs 775:781 The cross-linked CD-MOFs 758:781 The cross-linked CD-MOFs (LCD-MOFs) 758:792 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 5 58 link cross-linked 762:773 arg1 ligand 840:845 ligand 840:845 ligand 840:845 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 6 59 theme LCD-MOFs 941:948 arg1 performance 950:960 The LCD-MOFs performance 937:960 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples 937:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 9 60 theme unique 1716:1721 arg1 sites 1739:1743 344 unique N-glycosylation sites 1712:1743 344 unique N-glycosylation sites that mapped to 290 different glycoproteins 1712:1786 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 2 61 theme Hydrophilic 358:368 arg1 HILIC 398:402 HILIC 398:402 HILIC 398:402 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 2 61 theme Hydrophilic 358:368 arg1 chromatography 382:395 Hydrophilic interaction chromatography 358:395 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 8 62 theme detection 1474:1482 arg1 limit 1484:1488 the lower detection limit 1464:1488 the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG 1464:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 62 theme detection 1474:1482 arg1 fmol 1495:1498 3.3 fmol 1491:1498 3.3 fmol 1491:1498 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 4 63 theme hydrophilic 670:680 arg1 frameworks 696:705 hydrophilic metal-organic frameworks 670:705 hydrophilic metal-organic frameworks (MOFs) 670:712 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 63 theme hydrophilic 670:680 arg1 matrix 750:755 a HILIC matrix 742:755 a HILIC matrix 742:755 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 63 theme hydrophilic 670:680 arg1 MOFs 708:711 MOFs 708:711 MOFs 708:711 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 3 64 theme matrices 645:652 arg1 selectivity 624:634 the medium selectivity 613:634 the medium selectivity of HILIC matrices 613:652 However, the application of these methods is limited by the medium selectivity of HILIC matrices. 25110774 5 65 theme superior 889:896 arg1 hydrophilicity 898:911 superior hydrophilicity 889:911 superior hydrophilicity 889:911 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 6 66 theme complex 1017:1023 arg1 samples 1036:1042 the complex biological samples 1013:1042 the complex biological samples 1013:1042 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 8 67 theme recovery 1521:1528 arg1 yield 1530:1534 a satisfactory recovery yield 1506:1534 a satisfactory recovery yield (84-103%) 1506:1544 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 8 67 theme recovery 1521:1528 arg1 % 1543:1543 84-103% 1537:1543 84-103% 1537:1543 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 4 68 theme HILIC 744:748 arg1 frameworks 696:705 hydrophilic metal-organic frameworks 670:705 hydrophilic metal-organic frameworks (MOFs) 670:712 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 68 theme HILIC 744:748 arg1 matrix 750:755 a HILIC matrix 742:755 a HILIC matrix 742:755 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 10 69 theme HILIC 1877:1881 arg1 matrix 1883:1888 a HILIC matrix 1875:1888 a HILIC matrix 1875:1888 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 8 70 gly glycopeptide 1550:1561 arg2 glycopeptide 1550:1561 glycopeptide enrichment 1550:1572 glycopeptide enrichment from a digest of IgG 1550:1593 In addition, the resulting LCD-MOFs demonstrated the lower detection limit (3.3 fmol) with a satisfactory recovery yield (84-103%) for glycopeptide enrichment from a digest of IgG. 25110774 6 71 theme G 1110:1110 arg1 mixture 1078:1084 a digested mixture 1067:1084 a digested mixture of human immunoglobulin G (IgG) that was used as standard samples 1067:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 7 72 theme glycopeptides 1291:1303 arg1 signal 1281:1286 the signal 1277:1286 the signal of glycopeptides 1277:1303 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 6 73 theme human 1089:1093 arg1 samples 1144:1150 standard samples 1135:1150 standard samples 1135:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 73 theme human 1089:1093 arg1 IgG 1113:1115 IgG 1113:1115 IgG 1113:1115 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 73 theme human 1089:1093 arg1 G 1110:1110 human immunoglobulin G 1089:1110 human immunoglobulin G (IgG) that was used as standard samples 1089:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 9 74 theme MS 1816:1817 arg1 run 1819:1821 a single MS run 1807:1821 a single MS run 1807:1821 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 5 75 theme cross-linked 762:773 arg1 LCD-MOFs 784:791 LCD-MOFs 784:791 LCD-MOFs 784:791 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 5 75 theme cross-linked 762:773 arg1 CD-MOFs 775:781 The cross-linked CD-MOFs 758:781 The cross-linked CD-MOFs (LCD-MOFs) 758:792 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 5 75 theme cross-linked 762:773 arg1 ligand 840:845 ligand 840:845 ligand 840:845 The cross-linked CD-MOFs (LCD-MOFs) were facilely prepared with γ-cyclodextrin as ligand and possessed nano-sized cubic structure, superior hydrophilicity, and bio-compatibility. 25110774 7 76 theme IgG 1403:1405 arg1 digest 1407:1412 IgG digest 1403:1412 IgG digest 1403:1412 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 1 77 gly glycoproteins 275:287 arg1 glycoproteins 275:287 glycoproteins 275:287 glycoproteins using mass spectrometry (MS) 275:316 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 1 78 gly glycopeptides 136:148 arg2 glycopeptides 136:148 glycopeptides 136:148 glycopeptides 136:148 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 6 79 theme standard 1135:1142 arg1 samples 1144:1150 standard samples 1135:1150 standard samples 1135:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 79 theme standard 1135:1142 arg1 G 1110:1110 human immunoglobulin G 1089:1110 human immunoglobulin G (IgG) that was used as standard samples 1089:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 10 80 theme low-abundant 1951:1962 arg1 glycopeptides 1964:1976 identifying and enriching low-abundant glycopeptides 1925:1976 identifying and enriching low-abundant glycopeptides 1925:1976 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 6 81 used used 1127:1130 arg2 G 1110:1110 human immunoglobulin G 1089:1110 human immunoglobulin G (IgG) that was used as standard samples 1089:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 81 used used 1127:1130 arg2 samples 1144:1150 standard samples 1135:1150 standard samples 1135:1150 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 6 81 used used 1127:1130 arg2 IgG 1113:1115 IgG 1113:1115 IgG 1113:1115 The LCD-MOFs performance for the selective enrichment of glycopeptides from the complex biological samples were investigated with a digested mixture of human immunoglobulin G (IgG) that was used as standard samples. 25110774 1 82 theme Selective 112:120 arg1 enrichment 122:131 Selective enrichment 112:131 Selective enrichment of glycopeptides from complicated biological samples 112:184 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 7 83 gly glycopeptides 1291:1303 arg2 glycopeptides 1291:1303 glycopeptides 1291:1303 glycopeptides 1291:1303 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 0 84 theme glycopeptides 24:36 arg1 enrichment 10:19 Efficient enrichment 0:19 Efficient enrichment of glycopeptides 0:36 Efficient enrichment of glycopeptides using metal-organic frameworks by hydrophilic interaction chromatography. 25110774 4 85 used used 734:737 arg2 matrix 750:755 a HILIC matrix 742:755 a HILIC matrix 742:755 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 85 used used 734:737 arg2 MOFs 708:711 MOFs 708:711 MOFs 708:711 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 4 85 used used 734:737 arg2 frameworks 696:705 hydrophilic metal-organic frameworks 670:705 hydrophilic metal-organic frameworks (MOFs) 670:712 In this study, hydrophilic metal-organic frameworks (MOFs) were fabricated and used as a HILIC matrix. 25110774 2 86 theme glycopeptides 468:480 arg1 isolation 455:463 selective isolation 445:463 selective isolation of glycopeptides 445:480 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 7 87 theme ion 1214:1216 arg1 suppression 1218:1228 ion suppression 1214:1228 ion suppression 1214:1228 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 2 88 gly glycopeptides 525:537 arg2 glycopeptides 525:537 glycopeptides 525:537 glycopeptides 525:537 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 2 89 theme glycopeptides 525:537 arg1 glycan 515:520 the glycan 511:520 the glycan of glycopeptides and the matrices 511:554 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 1 90 dep structure 232:240 arg1 the 228:230 the 228:230 the 228:230 Selective enrichment of glycopeptides from complicated biological samples is critical for glycoproteomics to obtain the structure and glycosylation information of glycoproteins using mass spectrometry (MS), which still remains a great challenge. 25110774 7 91 gly glycopeptides 1357:1369 arg2 glycopeptides 1357:1369 twenty glycopeptides 1350:1369 twenty glycopeptides 1350:1369 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 10 92 theme identifying 1925:1935 arg1 glycopeptides 1964:1976 identifying and enriching low-abundant glycopeptides 1925:1976 identifying and enriching low-abundant glycopeptides 1925:1976 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 7 93 gly non-glycopeptides 1188:1204 arg2 non-glycopeptides 1188:1204 the non-glycopeptides 1184:1204 the non-glycopeptides causing ion suppression to the glycopeptides 1184:1249 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 7 94 gly glycopeptides 1237:1249 arg2 glycopeptides 1237:1249 the glycopeptides 1233:1249 the glycopeptides 1233:1249 In the selectivity assessment, the non-glycopeptides causing ion suppression to the glycopeptides were effectively removed, the signal of glycopeptides were enhanced significantly by LCD-MOFs, and twenty glycopeptides were identified with 67 fmol of IgG digest. 25110774 2 95 theme interaction 370:380 arg1 HILIC 398:402 HILIC 398:402 HILIC 398:402 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 2 95 theme interaction 370:380 arg1 chromatography 382:395 Hydrophilic interaction chromatography 358:395 Hydrophilic interaction chromatography (HILIC)-based strategies 358:420 Hydrophilic interaction chromatography (HILIC)-based strategies have been proposed for selective isolation of glycopeptides via the interactions between the glycan of glycopeptides and the matrices. 25110774 9 96 theme glycopeptides 1676:1688 arg1 enrichment 1662:1671 the selective enrichment 1648:1671 the selective enrichment of glycopeptides from mouse liver 1648:1705 Furthermore, a promising protocol was developed for the selective enrichment of glycopeptides from mouse liver, and 344 unique N-glycosylation sites that mapped to 290 different glycoproteins were identified in a single MS run. 25110774 10 97 contain have 1900:1903 arg1 LCD-MOFs 1891:1898 LCD-MOFs 1891:1898 LCD-MOFs 1891:1898 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 25110774 10 97 contain have 1900:1903 arg2 potential 1911:1919 great potential 1905:1919 great potential for identifying and enriching low-abundant glycopeptides in complex biological samples 1905:2006 The results clearly demonstrated that when used in a HILIC matrix, LCD-MOFs have great potential for identifying and enriching low-abundant glycopeptides in complex biological samples. 24451549 7 0 from significance 1365:1376 arg1 pathogenesis 1405:1416 pathogenesis 1405:1416 pathogenesis 1405:1416 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 3 1 theme BgAg-binding 665:676 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 1 theme BgAg-binding 665:676 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 1 theme BgAg-binding 665:676 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 7 2 theme O-glycosylation 1386:1400 arg1 significance 1365:1376 the significance 1361:1376 the significance of this O-glycosylation in pathogenesis 1361:1416 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 0 3 theme protein-mediated 76:91 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 3 theme protein-mediated 76:91 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 4 4 theme flagellin 795:803 arg1 proteins 805:812 previously only flagellin proteins 779:812 previously only flagellin proteins 779:812 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 5 from jejuni 852:857 arg1 O-glycosylated 831:844 O-glycosylated 831:844 O-glycosylated 831:844 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 6 6 from effect 1088:1093 arg1 conformation 1102:1113 the conformation 1098:1113 the conformation of MOMP 1098:1121 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 2 7 theme C. 508:509 arg1 jejuni 511:516 C. jejuni 508:516 C. jejuni 508:516 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 0 8 theme human 105:109 arg1 antigens 129:136 human histo-blood group antigens 105:136 human histo-blood group antigens 105:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 7 9 theme biofilm 1230:1236 arg1 formation 1238:1246 biofilm formation 1230:1246 biofilm formation 1230:1246 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 0 10 link O-linked 8:15 arg1 glycan 17:22 A novel O-linked glycan 0:22 A novel O-linked glycan 0:22 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 3 11 theme C. 690:691 arg1 jejuni 693:698 C. jejuni NCTC11168 690:708 C. jejuni NCTC11168 690:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 6 12 theme BgAg-binding 1142:1153 arg1 capacity 1155:1162 BgAg-binding capacity 1142:1162 BgAg-binding capacity 1142:1162 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 0 13 theme group 123:127 arg1 antigens 129:136 human histo-blood group antigens 105:136 human histo-blood group antigens 105:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 7 14 theme chickens 1327:1334 arg1 colonization 1311:1322 the optimal colonization 1299:1322 the optimal colonization of chickens by C. jejuni 1299:1347 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 2 15 theme human 414:418 arg1 BgAgs 448:452 BgAgs 448:452 BgAgs 448:452 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 2 15 theme human 414:418 arg1 antigens 438:445 human histo-blood group antigens 414:445 human histo-blood group antigens (BgAgs) 414:453 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 0 16 theme histo-blood 111:121 arg1 antigens 129:136 human histo-blood group antigens 105:136 human histo-blood group antigens 105:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 7 17 theme optimal 1303:1309 arg1 colonization 1311:1322 the optimal colonization 1299:1322 the optimal colonization of chickens by C. jejuni 1299:1347 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 1 18 theme important 191:199 arg1 cause 201:205 an important cause 188:205 an important cause of human foodborne gastroenteritis 188:240 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 1 18 theme important 191:199 arg1 jejuni 178:183 Campylobacter jejuni 164:183 Campylobacter jejuni 164:183 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 0 19 theme novel 2:6 arg1 glycan 17:22 A novel O-linked glycan 0:22 A novel O-linked glycan 0:22 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 4 20 gly O-glycosylated 751:764 arg1 O-glycosylated 751:764 O-glycosylated 751:764 O-glycosylated 751:764 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 20 gly O-glycosylated 751:764 arg1 MOMP 730:733 the MOMP 726:733 the MOMP 726:733 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 20 gly O-glycosylated 751:764 arg2 268 773:775 268 773:775 268 773:775 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 20 gly O-glycosylated 751:764 arg2 Thr 769:771 Thr 769:771 Thr(268) 769:776 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 5 21 theme MOMP 876:879 arg1 268 885:887 268 885:887 268 885:887 Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. 24451549 5 21 theme MOMP 876:879 arg1 Thr 881:883 MOMP Thr 876:883 MOMP Thr(268) 876:888 Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. 24451549 6 22 theme β1-3 982:985 arg1 moiety 950:955 The O-glycan moiety 937:955 The O-glycan moiety 937:955 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 22 theme β1-3 982:985 arg1 -GalNAcα1-Thr 1013:1025 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 1 23 theme poor 293:296 arg1 understanding 298:310 a poor understanding 291:310 a poor understanding of the complex interactions between host and pathogen 291:364 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 2 24 theme human 521:525 arg1 mucosa 538:543 human intestinal mucosa 521:543 human intestinal mucosa 521:543 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 5 25 theme Thr 881:883 arg1 Substitution 860:871 Substitution 860:871 Substitution of MOMP Thr(268) 860:888 Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. 24451549 3 26 theme membrane 622:629 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 26 theme membrane 622:629 arg1 MOMP 640:643 MOMP 640:643 MOMP 640:643 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 26 theme membrane 622:629 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 26 theme membrane 622:629 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 7 27 theme Caco-2 1264:1269 arg1 cells 1271:1275 Caco-2 cells 1264:1275 Caco-2 cells 1264:1275 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 1 28 theme human 210:214 arg1 gastroenteritis 226:240 human foodborne gastroenteritis 210:240 human foodborne gastroenteritis 210:240 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 0 29 theme O-linked 8:15 arg1 glycan 17:22 A novel O-linked glycan 0:22 A novel O-linked glycan 0:22 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 3 30 theme outer 616:620 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 30 theme outer 616:620 arg1 MOMP 640:643 MOMP 640:643 MOMP 640:643 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 30 theme outer 616:620 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 30 theme outer 616:620 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 1 31 theme foodborne 216:224 arg1 gastroenteritis 226:240 human foodborne gastroenteritis 210:240 human foodborne gastroenteritis 210:240 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 0 32 theme Campylobacter 34:46 arg1 colonization 150:161 chicken colonization 142:161 chicken colonization 142:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 32 theme Campylobacter 34:46 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 32 theme Campylobacter 34:46 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 2 33 theme group 432:436 arg1 BgAgs 448:452 BgAgs 448:452 BgAgs 448:452 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 2 33 theme group 432:436 arg1 antigens 438:445 human histo-blood group antigens 414:445 human histo-blood group antigens (BgAgs) 414:453 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 3 34 theme subunit 579:585 arg1 FlaA 596:599 FlaA 596:599 FlaA 596:599 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 34 theme subunit 579:585 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 34 theme subunit 579:585 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 34 theme subunit 579:585 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 35 from adhesins 678:685 arg1 jejuni 693:698 C. jejuni NCTC11168 690:708 C. jejuni NCTC11168 690:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 1 36 theme gastroenteritis 226:240 arg1 cause 201:205 an important cause 188:205 an important cause of human foodborne gastroenteritis 188:240 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 1 36 theme gastroenteritis 226:240 arg1 jejuni 178:183 Campylobacter jejuni 164:183 Campylobacter jejuni 164:183 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 6 37 theme -GalNAc 987:993 arg1 moiety 950:955 The O-glycan moiety 937:955 The O-glycan moiety 937:955 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 37 theme -GalNAc 987:993 arg1 -GalNAcα1-Thr 1013:1025 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 1 38 theme complex 319:325 arg1 interactions 327:338 the complex interactions 315:338 the complex interactions between host and pathogen 315:364 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 0 39 theme chicken 142:148 arg1 colonization 150:161 chicken colonization 142:161 chicken colonization 142:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 39 theme chicken 142:148 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 1 40 dep strategies 243:252 arg1 prevent 257:263 prevent 257:263 to prevent infection 254:273 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 0 41 dep jejuni 48:53 arg1 colonization 150:161 chicken colonization 142:161 chicken colonization 142:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 41 dep jejuni 48:53 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 41 dep jejuni 48:53 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 2 42 theme histo-blood 420:430 arg1 BgAgs 448:452 BgAgs 448:452 BgAgs 448:452 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 2 42 theme histo-blood 420:430 arg1 antigens 438:445 human histo-blood group antigens 414:445 human histo-blood group antigens (BgAgs) 414:453 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 0 43 theme major 55:59 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 43 theme major 55:59 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 1 44 theme interactions 327:338 arg1 understanding 298:310 a poor understanding 291:310 a poor understanding of the complex interactions between host and pathogen 291:364 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 7 45 gly Glycosylation 1165:1177 arg2 268 1194:1196 268 1194:1196 268 1194:1196 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 7 45 gly Glycosylation 1165:1177 arg2 Thr 1190:1192 Thr 1190:1192 Thr(268) 1190:1197 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 7 45 gly Glycosylation 1165:1177 arg1 MOMP 1182:1185 MOMP 1182:1185 MOMP 1182:1185 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 3 46 theme major 610:614 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 46 theme major 610:614 arg1 MOMP 640:643 MOMP 640:643 MOMP 640:643 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 46 theme major 610:614 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 46 theme major 610:614 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 47 theme major 564:568 arg1 FlaA 596:599 FlaA 596:599 FlaA 596:599 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 47 theme major 564:568 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 47 theme major 564:568 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 47 theme major 564:568 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 6 48 theme -GalNAc 1000:1006 arg1 moiety 950:955 The O-glycan moiety 937:955 The O-glycan moiety 937:955 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 48 theme -GalNAc 1000:1006 arg1 -GalNAcα1-Thr 1013:1025 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 2 49 theme Previous 367:374 arg1 work 376:379 Previous work 367:379 Previous work 367:379 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 2 50 theme intestinal 527:536 arg1 mucosa 538:543 human intestinal mucosa 521:543 human intestinal mucosa 521:543 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 2 51 theme jejuni 511:516 arg1 binding 497:503 the binding 493:503 the binding of C. jejuni to human intestinal mucosa 493:543 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 3 52 theme flagella 570:577 arg1 FlaA 596:599 FlaA 596:599 FlaA 596:599 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 52 theme flagella 570:577 arg1 protein 587:593 the major flagella subunit protein 560:593 the major flagella subunit protein (FlaA) 560:600 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 52 theme flagella 570:577 arg1 protein 631:637 the major outer membrane protein 606:637 the major outer membrane protein (MOMP) 606:644 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 3 52 theme flagella 570:577 arg1 adhesins 678:685 BgAg-binding adhesins 665:685 BgAg-binding adhesins in C. jejuni NCTC11168 665:708 Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168. 24451549 4 53 theme C. 849:850 arg1 jejuni 852:857 C. jejuni 849:857 C. jejuni 849:857 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 5 54 theme reduced 911:917 arg1 binding 919:925 significantly reduced binding 897:925 significantly reduced binding to BgAgs 897:934 Substitution of MOMP Thr(268) led to significantly reduced binding to BgAgs. 24451549 4 55 from Thr 769:771 arg1 O-glycosylated 751:764 O-glycosylated 751:764 O-glycosylated 751:764 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 55 from Thr 769:771 arg1 MOMP 730:733 the MOMP 726:733 the MOMP 726:733 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 7 56 theme C. 1339:1340 arg1 jejuni 1342:1347 C. jejuni 1339:1347 C. jejuni 1339:1347 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 7 57 from Thr 1190:1192 arg1 Glycosylation 1165:1177 Glycosylation 1165:1177 Glycosylation of MOMP at Thr(268) 1165:1197 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 4 58 from O-glycosylated 831:844 arg1 jejuni 852:857 C. jejuni 849:857 C. jejuni 849:857 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 6 59 theme notable 1080:1086 arg1 effect 1088:1093 a notable effect 1078:1093 a notable effect on the conformation of MOMP 1078:1121 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 4 60 gly O-glycosylated 831:844 arg1 jejuni 852:857 C. jejuni 849:857 C. jejuni 849:857 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 60 gly O-glycosylated 831:844 arg1 proteins 805:812 previously only flagellin proteins 779:812 previously only flagellin proteins 779:812 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 4 60 gly O-glycosylated 831:844 arg2 proteins 805:812 previously only flagellin proteins 779:812 previously only flagellin proteins 779:812 Significantly, the MOMP was shown to be O-glycosylated at Thr(268); previously only flagellin proteins were known to be O-glycosylated in C. jejuni. 24451549 6 61 theme O-glycan 941:948 arg1 -GalNAcα1-Thr 1013:1025 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 61 theme O-glycan 941:948 arg1 moiety 950:955 The O-glycan moiety 937:955 The O-glycan moiety 937:955 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 0 62 theme membrane 67:74 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 62 theme membrane 67:74 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 7 63 theme MOMP 1182:1185 arg1 Glycosylation 1165:1177 Glycosylation 1165:1177 Glycosylation of MOMP at Thr(268) 1165:1197 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 7 64 theme cell-to-cell 1208:1219 arg1 binding 1221:1227 cell-to-cell binding 1208:1227 cell-to-cell binding 1208:1227 Glycosylation of MOMP at Thr(268) promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni, confirming the significance of this O-glycosylation in pathogenesis. 24451549 6 65 theme Gal 978:980 arg1 moiety 950:955 The O-glycan moiety 937:955 The O-glycan moiety 937:955 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 65 theme Gal 978:980 arg1 -GalNAcα1-Thr 1013:1025 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268) 978:1030 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 2 66 theme C. 393:394 arg1 jejuni 396:401 C. jejuni 393:401 C. jejuni 393:401 Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. 24451549 6 67 contain has 1074:1076 arg2 effect 1088:1093 a notable effect 1078:1093 a notable effect on the conformation of MOMP 1078:1121 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 6 67 contain has 1074:1076 arg1 O-glycosylation 1058:1072 O-glycosylation 1058:1072 O-glycosylation 1058:1072 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 0 68 theme outer 61:65 arg1 adhesion 93:100 major outer membrane protein-mediated adhesion 55:100 major outer membrane protein-mediated adhesion to human histo-blood group antigens 55:136 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 0 68 theme outer 61:65 arg1 jejuni 48:53 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization 34:161 A novel O-linked glycan modulates Campylobacter jejuni major outer membrane protein-mediated adhesion to human histo-blood group antigens and chicken colonization. 24451549 6 69 theme MOMP 1118:1121 arg1 conformation 1102:1113 the conformation 1098:1113 the conformation of MOMP 1098:1121 The O-glycan moiety was characterized as Gal(β1-3)-GalNAc(β1-4)-GalNAc(β1-4)-GalNAcα1-Thr(268); modelling suggested that O-glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. 24451549 1 70 theme Campylobacter 164:176 arg1 cause 201:205 an important cause 188:205 an important cause of human foodborne gastroenteritis 188:240 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 24451549 1 70 theme Campylobacter 164:176 arg1 jejuni 178:183 Campylobacter jejuni 164:183 Campylobacter jejuni 164:183 Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. 28230186 0 0 theme cognate 73:79 arg1 receptor 90:97 its cognate cellular receptor 69:97 its cognate cellular receptor 69:97 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 0 0 theme cognate 73:79 arg1 FCMR 63:66 FCMR 63:66 FCMR 63:66 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 6 1 with interaction 803:813 arg1 IgM 820:822 IgM 820:822 IgM 820:822 We generated a homology-based structure for hFCMR and used molecular dynamic simulations to show how this interaction with IgM may occur. 28230186 2 2 from sites 302:306 arg1 chain 317:321 the μ chain 311:321 the μ chain 311:321 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 2 3 theme μ 315:315 arg1 chain 317:321 the μ chain 311:321 the μ chain 311:321 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 1 4 theme viruses 227:233 arg1 neutralization 196:209 the neutralization 192:209 the neutralization of bacteria and viruses 192:233 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 5 5 theme hFCMR 609:613 arg1 IgM 588:590 IgM 588:590 IgM 588:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 5 theme hFCMR 609:613 arg1 target 599:604 the target 595:604 the target of hFCMR 595:613 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 5 theme hFCMR 609:613 arg1 domain 578:583 the Cμ4 domain 570:583 the Cμ4 domain of IgM 570:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 4 6 theme Fcμ 521:523 arg1 hFCMR 535:539 hFCMR 535:539 hFCMR 535:539 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 4 6 theme Fcμ 521:523 arg1 receptor 525:532 human Fcμ receptor 515:532 the human Fcμ receptor (hFCMR) 511:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 2 7 gly glycosylated 257:268 arg1 IgM 242:244 Human IgM 236:244 Human IgM 236:244 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 3 8 gly Glycosylation 347:359 arg1 IgG 364:366 IgG 364:366 IgG 364:366 Glycosylation of IgG is known to modulate the effector functions of Fcγ receptors. 28230186 4 9 theme human 515:519 arg1 hFCMR 535:539 hFCMR 535:539 hFCMR 535:539 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 4 9 theme human 515:519 arg1 receptor 525:532 human Fcμ receptor 515:532 the human Fcμ receptor (hFCMR) 511:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 3 10 theme Fcγ 415:417 arg1 receptors 419:427 Fcγ receptors 415:427 Fcγ receptors 415:427 Glycosylation of IgG is known to modulate the effector functions of Fcγ receptors. 28230186 1 11 theme immune 144:149 arg1 responses 151:159 immune responses 144:159 immune responses 144:159 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 6 12 used used 751:754 arg2 We 697:698 We 697:698 We 697:698 We generated a homology-based structure for hFCMR and used molecular dynamic simulations to show how this interaction with IgM may occur. 28230186 6 13 theme dynamic 766:772 arg1 simulations 774:784 molecular dynamic simulations 756:784 molecular dynamic simulations 756:784 We generated a homology-based structure for hFCMR and used molecular dynamic simulations to show how this interaction with IgM may occur. 28230186 3 14 theme IgG 364:366 arg1 Glycosylation 347:359 Glycosylation 347:359 Glycosylation of IgG 347:366 Glycosylation of IgG is known to modulate the effector functions of Fcγ receptors. 28230186 6 15 theme molecular 756:764 arg1 simulations 774:784 molecular dynamic simulations 756:784 molecular dynamic simulations 756:784 We generated a homology-based structure for hFCMR and used molecular dynamic simulations to show how this interaction with IgM may occur. 28230186 2 16 theme N-linked 286:293 arg1 sites 302:306 five N-linked glycan sites 281:306 five N-linked glycan sites on the μ chain 281:321 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 0 17 theme Glycan-independent 0:17 arg1 binding 19:25 Glycan-independent binding 0:25 Glycan-independent binding 0:25 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 2 18 from one 327:329 arg1 J-chain 338:344 the J-chain 334:344 the J-chain 334:344 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 7 19 from function 868:875 arg1 proliferation 892:904 the proliferation 888:904 the proliferation of T cells 888:915 Finally, we reveal an inhibitory function for IgM in the proliferation of T cells. 28230186 4 20 theme glycosylation 479:491 arg1 effect 469:474 the effect 465:474 the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR) 465:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 1 21 theme important 174:182 arg1 role 184:187 an important role 171:187 an important role 171:187 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 0 22 theme human 50:54 arg1 IgM 56:58 human IgM 50:58 human IgM 50:58 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 4 23 theme IgM 496:498 arg1 binding 500:506 IgM binding 496:506 IgM binding to the human Fcμ receptor (hFCMR) 496:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 3 24 theme effector 393:400 arg1 functions 402:410 the effector functions 389:410 the effector functions of Fcγ receptors 389:427 Glycosylation of IgG is known to modulate the effector functions of Fcγ receptors. 28230186 4 25 from effect 469:474 arg1 binding 500:506 IgM binding 496:506 IgM binding to the human Fcμ receptor (hFCMR) 496:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 5 26 theme IgM 661:663 arg1 binding 630:636 binding 630:636 binding 630:636 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 26 theme IgM 661:663 arg1 internalization 642:656 internalization 642:656 internalization 642:656 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 4 27 gly glycosylation 479:491 arg1 IgM 496:498 IgM binding 496:506 IgM binding to the human Fcμ receptor (hFCMR) 496:540 In contrast, little is known about the effect of glycosylation on IgM binding to the human Fcμ receptor (hFCMR). 28230186 5 28 theme Cμ4 574:576 arg1 IgM 588:590 IgM 588:590 IgM 588:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 28 theme Cμ4 574:576 arg1 target 599:604 the target 595:604 the target of hFCMR 595:613 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 28 theme Cμ4 574:576 arg1 domain 578:583 the Cμ4 domain 570:583 the Cμ4 domain of IgM 570:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 0 29 theme IgM 56:58 arg1 internalization 31:45 internalization 31:45 internalization 31:45 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 0 29 theme IgM 56:58 arg1 binding 19:25 Glycan-independent binding 0:25 Glycan-independent binding 0:25 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 2 30 link N-linked 286:293 arg1 sites 302:306 five N-linked glycan sites 281:306 five N-linked glycan sites on the μ chain 281:321 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 7 31 theme cells 911:915 arg1 proliferation 892:904 the proliferation 888:904 the proliferation of T cells 888:915 Finally, we reveal an inhibitory function for IgM in the proliferation of T cells. 28230186 6 32 theme homology-based 712:725 arg1 structure 727:735 a homology-based structure 710:735 a homology-based structure for hFCMR 710:745 We generated a homology-based structure for hFCMR and used molecular dynamic simulations to show how this interaction with IgM may occur. 28230186 2 33 theme Human 236:240 arg1 IgM 242:244 Human IgM 236:244 Human IgM 236:244 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 1 34 theme first 111:115 arg1 antibody 117:124 the first antibody 107:124 the first antibody to be produced in immune responses 107:159 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 1 34 theme first 111:115 arg1 IgM 100:102 IgM 100:102 IgM 100:102 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 5 35 theme IgM 588:590 arg1 IgM 588:590 IgM 588:590 IgM 588:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 35 theme IgM 588:590 arg1 target 599:604 the target 595:604 the target of hFCMR 595:613 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 5 35 theme IgM 588:590 arg1 domain 578:583 the Cμ4 domain 570:583 the Cμ4 domain of IgM 570:590 In this study, we identify the Cμ4 domain of IgM as the target of hFCMR, and show that binding and internalization of IgM by hFCMR is glycan-independent. 28230186 7 36 theme inhibitory 857:866 arg1 function 868:875 an inhibitory function 854:875 an inhibitory function for IgM in the proliferation of T cells 854:915 Finally, we reveal an inhibitory function for IgM in the proliferation of T cells. 28230186 2 37 theme glycan 295:300 arg1 sites 302:306 five N-linked glycan sites 281:306 five N-linked glycan sites on the μ chain 281:321 Human IgM is heavily glycosylated, featuring five N-linked glycan sites on the μ chain and one on the J-chain. 28230186 3 38 theme receptors 419:427 arg1 functions 402:410 the effector functions 389:410 the effector functions of Fcγ receptors 389:427 Glycosylation of IgG is known to modulate the effector functions of Fcγ receptors. 28230186 7 39 theme T 909:909 arg1 cells 911:915 T cells 909:915 T cells 909:915 Finally, we reveal an inhibitory function for IgM in the proliferation of T cells. 28230186 1 40 theme bacteria 214:221 arg1 neutralization 196:209 the neutralization 192:209 the neutralization of bacteria and viruses 192:233 IgM is the first antibody to be produced in immune responses and plays an important role in the neutralization of bacteria and viruses. 28230186 0 41 theme cellular 81:88 arg1 receptor 90:97 its cognate cellular receptor 69:97 its cognate cellular receptor 69:97 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 28230186 0 41 theme cellular 81:88 arg1 FCMR 63:66 FCMR 63:66 FCMR 63:66 Glycan-independent binding and internalization of human IgM to FCMR, its cognate cellular receptor. 29285644 10 0 theme biological 1764:1773 arg1 information 1775:1785 biological information 1764:1785 biological information as well as high sensitivity and reproducibility 1764:1833 From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery. 29285644 11 1 theme Graphical 1878:1886 arg1 abstract 1888:1895 Graphical abstract 1878:1895 Graphical abstract 1878:1895 Graphical abstract ᅟ. 29285644 4 2 with platform 870:877 arg1 reaction 902:909 a one-shot enzyme reaction 884:909 a one-shot enzyme reaction 884:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 0 3 theme haptoglobin 87:97 arg1 analysis 69:76 targeted glycoproteomic analysis 45:76 targeted glycoproteomic analysis of serum haptoglobin 45:97 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 5 4 theme multispecific 989:1001 arg1 proteolysis 1003:1013 multispecific proteolysis 989:1013 multispecific proteolysis 989:1013 Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. 29285644 7 5 gly glycopeptides 1282:1294 arg2 each 1297:1300 each 1297:1300 each 1297:1300 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 7 5 gly glycopeptides 1282:1294 arg2 glycopeptides 1282:1294 Ninety-six glycopeptides 1271:1294 Ninety-six glycopeptides 1271:1294 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 4 6 theme enzyme 895:900 arg1 reaction 902:909 a one-shot enzyme reaction 884:909 a one-shot enzyme reaction 884:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 4 7 theme target-protein-specific 793:815 arg1 it 785:786 it 785:786 it 785:786 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 4 7 theme target-protein-specific 793:815 arg1 platform 870:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform 791:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction 791:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 6 8 theme chromatography-mass 1181:1199 arg1 spectrometry 1201:1212 nano liquid chromatography-mass spectrometry 1169:1212 nano liquid chromatography-mass spectrometry 1169:1212 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 5 9 gly glycopeptides 1041:1053 arg2 glycopeptides 1041:1053 site-specific glycopeptides 1027:1053 site-specific glycopeptides 1027:1053 Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. 29285644 5 10 theme site-specific 1027:1039 arg1 glycopeptides 1041:1053 site-specific glycopeptides 1027:1053 site-specific glycopeptides 1027:1053 Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. 29285644 3 11 with platform 548:555 arg1 approach 588:595 a targeted glycoproteomic approach 562:595 a targeted glycoproteomic approach for GC biomarker discovery 562:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 3 12 theme targeted 564:571 arg1 approach 588:595 a targeted glycoproteomic approach 562:595 a targeted glycoproteomic approach for GC biomarker discovery 562:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 2 13 theme aberrant 332:339 arg1 glycosylation 341:353 aberrant glycosylation 332:353 aberrant glycosylation 332:353 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 0 14 theme cancer 122:127 arg1 discovery 139:147 gastric cancer biomarker discovery 114:147 gastric cancer biomarker discovery 114:147 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 8 15 from Differences 1406:1416 arg1 abundance 1421:1429 abundance 1421:1429 abundance 1421:1429 Differences in abundance between the two groups were marked by particularly high magnitudes. 29285644 7 16 theme Ninety-six 1271:1280 arg1 each 1297:1300 each 1297:1300 each 1297:1300 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 7 16 theme Ninety-six 1271:1280 arg1 glycopeptides 1282:1294 Ninety-six glycopeptides 1271:1294 Ninety-six glycopeptides 1271:1294 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 6 17 theme mass 1252:1255 arg1 spectrometry 1257:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 4 18 theme glycosylation-site-specific 818:844 arg1 it 785:786 it 785:786 it 785:786 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 4 18 theme glycosylation-site-specific 818:844 arg1 platform 870:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform 791:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction 791:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 5 19 theme immunoaffinity 942:955 arg1 chromatography 957:970 immunoaffinity chromatography 942:970 immunoaffinity chromatography 942:970 Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. 29285644 2 20 theme glycosylation 412:424 arg1 localization 392:403 site-specific localization 378:403 site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use 378:481 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 2 20 theme glycosylation 412:424 arg1 challenge 506:514 an analytical challenge 492:514 an analytical challenge 492:514 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 0 21 theme gastric 114:120 arg1 discovery 139:147 gastric cancer biomarker discovery 114:147 gastric cancer biomarker discovery 114:147 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 2 22 theme analytical 495:504 arg1 challenge 506:514 an analytical challenge 492:514 an analytical challenge 492:514 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 2 22 theme analytical 495:504 arg1 localization 392:403 site-specific localization 378:403 site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use 378:481 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 9 23 theme curve 1624:1628 arg1 areas 1630:1634 receiver operating characteristic curve areas 1590:1634 receiver operating characteristic curve areas of 1.0 1590:1641 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 4 24 theme conventional 636:647 arg1 approach 658:665 the conventional glycomic approach 632:665 the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan 632:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 6 25 theme chromatography-tandem 1230:1250 arg1 spectrometry 1257:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 10 26 theme high 1798:1801 arg1 sensitivity 1803:1813 high sensitivity 1798:1813 high sensitivity 1798:1813 From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery. 29285644 9 27 theme high 1543:1546 arg1 changes 1571:1577 exceptionally high control-to-cancer fold changes 1529:1577 exceptionally high control-to-cancer fold changes 1529:1577 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 6 28 theme liquid 1223:1228 arg1 spectrometry 1257:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 4 29 theme glycan 724:729 arg1 profiling 702:710 untargeted mass spectrometric profiling 672:710 untargeted mass spectrometric profiling of released glycan 672:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 2 30 theme clinical 470:477 arg1 use 479:481 clinical use 470:481 clinical use 470:481 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 1 31 theme early 269:273 arg1 diagnosis 275:283 early diagnosis 269:283 early diagnosis 269:283 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 7 32 theme glycan/glycosite 1328:1343 arg1 pairing 1345:1351 a unique glycan/glycosite pairing 1319:1351 a unique glycan/glycosite pairing 1319:1351 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 3 33 theme GC 601:602 arg1 discovery 614:622 GC biomarker discovery 601:622 GC biomarker discovery 601:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 4 34 theme released 715:722 arg1 glycan 724:729 released glycan 715:729 released glycan 715:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 0 35 theme glycopeptides 27:39 arg1 Designation 0:10 Designation 0:10 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 0:148 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 9 36 theme control-to-cancer 1548:1564 arg1 changes 1571:1577 exceptionally high control-to-cancer fold changes 1529:1577 exceptionally high control-to-cancer fold changes 1529:1577 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 1 37 from difficulties 253:264 arg1 diagnosis 275:283 early diagnosis 269:283 early diagnosis 269:283 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 4 38 theme one-shot 886:893 arg1 reaction 902:909 a one-shot enzyme reaction 884:909 a one-shot enzyme reaction 884:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 3 39 theme glycoproteomic 573:586 arg1 approach 588:595 a targeted glycoproteomic approach 562:595 a targeted glycoproteomic approach for GC biomarker discovery 562:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 9 40 gly glycopeptides 1505:1517 arg2 glycopeptides 1505:1517 Three glycopeptides 1499:1517 Three glycopeptides 1499:1517 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 1 41 theme leading 184:190 arg1 causes 192:197 the leading causes 180:197 the leading causes of cancer-related death worldwide 180:231 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 0 42 theme fingerprint 15:25 arg1 glycopeptides 27:39 fingerprint glycopeptides 15:39 fingerprint glycopeptides 15:39 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 9 43 theme fold 1566:1569 arg1 changes 1571:1577 exceptionally high control-to-cancer fold changes 1529:1577 exceptionally high control-to-cancer fold changes 1529:1577 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 0 44 theme biomarker 129:137 arg1 discovery 139:147 gastric cancer biomarker discovery 114:147 gastric cancer biomarker discovery 114:147 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 2 45 theme accumulating 294:305 arg1 evidence 307:314 accumulating evidence 294:314 accumulating evidence indicating that aberrant glycosylation is associated with GC 294:375 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 9 46 theme perfect 1655:1661 arg1 discrimination 1663:1676 perfect discrimination 1655:1676 perfect discrimination between the two groups 1655:1699 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 1 47 theme causes 192:197 arg1 causes 192:197 the leading causes 180:197 the leading causes of cancer-related death worldwide 180:231 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 1 47 theme causes 192:197 arg1 one 173:175 one 173:175 one 173:175 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 0 48 theme targeted 45:52 arg1 analysis 69:76 targeted glycoproteomic analysis 45:76 targeted glycoproteomic analysis of serum haptoglobin 45:97 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 5 49 theme Serum 912:916 arg1 haptoglobin 918:928 Serum haptoglobin 912:928 Serum haptoglobin enriched by immunoaffinity chromatography 912:970 Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. 29285644 4 50 with approach 658:665 arg1 profiling 702:710 untargeted mass spectrometric profiling 672:710 untargeted mass spectrometric profiling of released glycan 672:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 6 51 theme nano 1218:1221 arg1 spectrometry 1257:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 nano liquid chromatography-tandem mass spectrometry 1218:1268 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 4 52 theme spectrometric 688:700 arg1 profiling 702:710 untargeted mass spectrometric profiling 672:710 untargeted mass spectrometric profiling of released glycan 672:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 6 53 theme nano 1169:1172 arg1 spectrometry 1201:1212 nano liquid chromatography-mass spectrometry 1169:1212 nano liquid chromatography-mass spectrometry 1169:1212 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 9 54 theme operating 1599:1607 arg1 curve 1624:1628 receiver operating characteristic curve 1590:1628 receiver operating characteristic curve areas of 1.0 1590:1641 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 7 55 gly glycan/glycosite 1328:1343 arg2 glycan/glycosite 1328:1343 a unique glycan/glycosite pairing 1319:1351 a unique glycan/glycosite pairing 1319:1351 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 9 56 theme 1.0 1639:1641 arg1 areas 1630:1634 receiver operating characteristic curve areas 1590:1634 receiver operating characteristic curve areas of 1.0 1590:1641 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 1 57 theme cancer-related 202:215 arg1 worldwide 223:231 cancer-related death worldwide 202:231 cancer-related death worldwide 202:231 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 10 58 theme biomarker 1857:1865 arg1 discovery 1867:1875 GC biomarker discovery 1854:1875 GC biomarker discovery 1854:1875 From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery. 29285644 6 59 theme liquid 1174:1179 arg1 spectrometry 1201:1212 nano liquid chromatography-mass spectrometry 1169:1212 nano liquid chromatography-mass spectrometry 1169:1212 Glycopeptides were identified and quantified by nano liquid chromatography-mass spectrometry and nano liquid chromatography-tandem mass spectrometry. 29285644 3 60 theme biomarker 604:612 arg1 discovery 614:622 GC biomarker discovery 601:622 GC biomarker discovery 601:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 8 61 theme high 1482:1485 arg1 magnitudes 1487:1496 particularly high magnitudes 1469:1496 particularly high magnitudes 1469:1496 Differences in abundance between the two groups were marked by particularly high magnitudes. 29285644 0 62 theme glycoproteomic 54:67 arg1 analysis 69:76 targeted glycoproteomic analysis 45:76 targeted glycoproteomic analysis of serum haptoglobin 45:97 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 7 63 theme cancer 1378:1383 arg1 samples 1397:1403 all cancer and control samples 1374:1403 samples 1397:1403 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 0 64 gly glycopeptides 27:39 arg2 glycopeptides 27:39 fingerprint glycopeptides 15:39 fingerprint glycopeptides 15:39 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 7 65 theme control 1389:1395 arg1 samples 1397:1403 all cancer and control samples 1374:1403 samples 1397:1403 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 4 66 theme structure-specific 851:868 arg1 it 785:786 it 785:786 it 785:786 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 4 66 theme structure-specific 851:868 arg1 platform 870:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform 791:877 a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction 791:909 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 10 67 theme GC 1854:1855 arg1 discovery 1867:1875 GC biomarker discovery 1854:1875 GC biomarker discovery 1854:1875 From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery. 29285644 7 68 theme unique 1321:1326 arg1 pairing 1345:1351 a unique glycan/glycosite pairing 1319:1351 a unique glycan/glycosite pairing 1319:1351 Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. 29285644 4 69 theme key 771:773 arg1 features 775:782 three key features 765:782 three key features 765:782 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 9 70 theme characteristic 1609:1622 arg1 curve 1624:1628 receiver operating characteristic curve 1590:1628 receiver operating characteristic curve areas of 1.0 1590:1641 Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. 29285644 1 71 theme death 217:221 arg1 worldwide 223:231 cancer-related death worldwide 202:231 cancer-related death worldwide 202:231 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 1 72 theme Gastric 150:156 arg1 GC 166:167 GC 166:167 GC 166:167 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 1 72 theme Gastric 150:156 arg1 cancer 158:163 Gastric cancer 150:163 Gastric cancer (GC) 150:168 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 1 73 theme worldwide 223:231 arg1 causes 192:197 the leading causes 180:197 the leading causes of cancer-related death worldwide 180:231 Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. 29285644 0 74 theme serum 81:85 arg1 haptoglobin 87:97 serum haptoglobin 81:97 serum haptoglobin 81:97 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 4 75 theme mass 683:686 arg1 profiling 702:710 untargeted mass spectrometric profiling 672:710 untargeted mass spectrometric profiling of released glycan 672:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 4 76 theme untargeted 672:681 arg1 profiling 702:710 untargeted mass spectrometric profiling 672:710 untargeted mass spectrometric profiling of released glycan 672:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 29285644 2 77 theme site-specific 378:390 arg1 localization 392:403 site-specific localization 378:403 site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use 378:481 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 2 77 theme site-specific 378:390 arg1 challenge 506:514 an analytical challenge 492:514 an analytical challenge 492:514 Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. 29285644 10 78 from results 1711:1717 arg1 useful 1843:1848 useful 1843:1848 useful 1843:1848 From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery. 29285644 0 79 dep Designation 0:10 arg1 insights 100:107 insights 100:107 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 0:148 Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery. 29285644 3 80 theme analytical 537:546 arg1 platform 548:555 an analytical platform 534:555 an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery 534:622 Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. 29285644 4 81 theme glycomic 649:656 arg1 approach 658:665 the conventional glycomic approach 632:665 the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan 632:729 Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. 28529241 0 0 theme N-Glycans 164:172 arg1 β4-Galactosylation 126:143 Decreased β4-Galactosylation 116:143 Decreased β4-Galactosylation of Highly Branched N-Glycans 116:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 4 1 theme glycoproteins 648:660 arg1 properties 676:685 malignant properties 666:685 malignant properties of A549 human lung cancer cell line 666:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 4 1 theme glycoproteins 648:660 arg1 N-glycosylation 629:643 N-glycosylation 629:643 N-glycosylation of glycoproteins 629:660 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 10 2 theme A549 1877:1880 arg1 cells 1882:1886 A549 cells 1877:1886 A549 cells 1877:1886 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 1 3 theme malignant 266:274 arg1 transformation 276:289 malignant transformation 266:289 malignant transformation of cells 266:298 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 4 4 gly glycoproteins 648:660 arg1 glycoproteins 648:660 glycoproteins 648:660 glycoproteins 648:660 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 4 5 theme malignant 666:674 arg1 properties 676:685 malignant properties 666:685 malignant properties of A549 human lung cancer cell line 666:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 9 6 from suppression 1660:1670 arg1 agar 1716:1719 soft agar 1711:1719 soft agar 1711:1719 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 9 6 from suppression 1660:1670 arg1 activity 1735:1742 migratory activity 1725:1742 migratory activity 1725:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 10 7 theme decreased 1900:1908 arg1 β4-galactosylation 1910:1927 the decreased β4-galactosylation 1896:1927 the decreased β4-galactosylation of highly branched N-glycans 1896:1956 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 6 8 theme adhesion 947:954 arg1 E-cadherin 966:975 E-cadherin 966:975 E-cadherin 966:975 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 6 8 theme adhesion 947:954 arg1 molecule 956:963 cell adhesion molecule 942:963 cell adhesion molecule 942:963 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 1 9 theme glycan 199:204 arg1 structures 206:215 the glycan structures 195:215 the glycan structures of cell surface proteins 195:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 4 10 theme cancer 706:711 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 0 11 theme Decreased 116:124 arg1 β4-Galactosylation 126:143 Decreased β4-Galactosylation 116:143 Decreased β4-Galactosylation of Highly Branched N-Glycans 116:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 10 12 theme branched 1939:1946 arg1 N-glycans 1948:1956 highly branched N-glycans 1932:1956 highly branched N-glycans 1932:1956 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 10 13 theme present 1784:1790 arg1 study 1792:1796 The present study 1780:1796 The present study 1780:1796 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 4 14 theme line 718:721 arg1 properties 676:685 malignant properties 666:685 malignant properties of A549 human lung cancer cell line 666:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 4 14 theme line 718:721 arg1 N-glycosylation 629:643 N-glycosylation 629:643 N-glycosylation of glycoproteins 629:660 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 7 15 theme β4-galactosyltransferase 1102:1125 arg1 β4GalT 1128:1133 β4-galactosyltransferase (β4GalT) 1 1102:1136 β4-galactosyltransferase (β4GalT) 1 1102:1136 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 1 16 theme surface 225:231 arg1 proteins 233:240 cell surface proteins 220:240 cell surface proteins 220:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 0 17 theme Branched 155:162 arg1 N-Glycans 164:172 Highly Branched N-Glycans 148:172 Highly Branched N-Glycans 148:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 9 18 theme Sp1-downregulated 1625:1641 arg1 cells 1643:1647 the Sp1-downregulated cells 1621:1647 the Sp1-downregulated cells 1621:1647 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 8 19 theme gene 1602:1605 arg1 expression 1576:1585 the reduced expression 1564:1585 the reduced expression of the β4GalT1 gene 1564:1605 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 5 20 theme Sp1-expression 760:773 arg1 level 775:779 level 775:779 level 775:779 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 7 21 theme gene 1077:1080 arg1 level 1093:1097 the gene expression level 1073:1097 the gene expression level of β4-galactosyltransferase (β4GalT) 1 1073:1136 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 9 22 from agar 1716:1719 arg1 suppression 1660:1670 the suppression 1656:1670 the suppression of the anchorage-independent growth in soft agar and migratory activity 1656:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 7 23 theme promoter 1341:1348 arg1 activity 1350:1357 the promoter activity 1337:1357 the promoter activity 1337:1357 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 24 theme Sp1-binding 1278:1288 arg1 promoter 1316:1323 the β4GalT1 gene promoter 1299:1323 the β4GalT1 gene promoter 1299:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 24 theme Sp1-binding 1278:1288 arg1 sites 1290:1294 the Sp1-binding sites 1274:1294 the Sp1-binding sites of the β4GalT1 gene promoter 1274:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 1 25 from changes 184:190 arg1 structures 206:215 the glycan structures 195:215 the glycan structures of cell surface proteins 195:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 0 26 theme Human 83:87 arg1 Line 106:109 A549 Human Lung Cancer Cell Line 78:109 A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 78:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 7 27 theme N-glycans 1051:1059 arg1 β4-galactosylation 1029:1046 decreased β4-galactosylation 1019:1046 decreased β4-galactosylation of N-glycans 1019:1059 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 1 28 theme Dramatic 175:182 arg1 changes 184:190 Dramatic changes 175:190 Dramatic changes in the glycan structures of cell surface proteins 175:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 0 29 theme Cancer 94:99 arg1 Line 106:109 A549 Human Lung Cancer Cell Line 78:109 A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 78:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 9 30 theme growth 1701:1706 arg1 suppression 1660:1670 the suppression 1656:1670 the suppression of the anchorage-independent growth in soft agar and migratory activity 1656:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 10 31 theme Sp1 1834:1836 arg1 downregulation 1816:1829 downregulation 1816:1829 downregulation of Sp1 1816:1836 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 0 32 theme Line 106:109 arg1 Properties 64:73 Malignant Properties 54:73 Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 54:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 9 33 theme soft 1711:1714 arg1 agar 1716:1719 soft agar 1711:1719 soft agar 1711:1719 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 0 34 theme A549 78:81 arg1 Line 106:109 A549 Human Lung Cancer Cell Line 78:109 A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 78:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 3 35 theme Transcription 452:464 arg1 Sp1 473:475 Transcription factor Sp1 452:475 Transcription factor Sp1 452:475 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 1 36 theme cells 294:298 arg1 transformation 276:289 malignant transformation 266:289 malignant transformation of cells 266:298 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 7 37 theme underlying 994:1003 arg1 mechanism 1005:1013 underlying mechanism 994:1013 underlying mechanism for decreased β4-galactosylation of N-glycans 994:1059 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 8 38 theme N-glycans 1511:1519 arg1 β4-galactosylation 1473:1490 the β4-galactosylation 1469:1490 the β4-galactosylation of highly branched N-glycans 1469:1519 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 0 39 theme Transcription 18:30 arg1 Sp1 39:41 Transcription Factor Sp1 18:41 Transcription Factor Sp1 18:41 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 6 40 theme cell 942:945 arg1 E-cadherin 966:975 E-cadherin 966:975 E-cadherin 966:975 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 6 40 theme cell 942:945 arg1 molecule 956:963 cell adhesion molecule 942:963 cell adhesion molecule 942:963 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 2 41 theme cancer 438:443 arg1 cells 445:449 cancer cells 438:449 cancer cells 438:449 Such changes are closely associated with the malignant properties of cancer cells. 28529241 0 42 theme Sp1 39:41 arg1 Downregulation 0:13 Downregulation 0:13 Downregulation of Transcription Factor Sp1 0:41 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 5 43 theme clone 813:817 arg1 clone 813:817 a control clone 803:817 a control clone 803:817 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 5 43 theme clone 813:817 arg1 % 798:798 50% 796:798 50% of a control clone 796:817 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 2 44 theme Such 369:372 arg1 changes 374:380 Such changes 369:380 Such changes 369:380 Such changes are closely associated with the malignant properties of cancer cells. 28529241 0 45 theme Malignant 54:62 arg1 Properties 64:73 Malignant Properties 54:73 Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 54:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 1 46 theme expression 326:335 arg1 levels 337:342 the altered expression levels 314:342 the altered expression levels of glycosyltransferases 314:366 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 6 47 theme N-glycans 912:920 arg1 β4-galactosylation 874:891 the β4-galactosylation 870:891 the β4-galactosylation of highly branched N-glycans 870:920 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 7 48 theme gene 1404:1407 arg1 expression 1409:1418 the gene expression 1400:1418 the gene expression 1400:1418 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 3 49 theme various 510:516 arg1 glycosyltransferases 538:557 glycosyltransferases 538:557 glycosyltransferases 538:557 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 3 49 theme various 510:516 arg1 molecules 518:526 various molecules 510:526 various molecules including glycosyltransferases 510:557 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 1 50 theme proteins 233:240 arg1 structures 206:215 the glycan structures 195:215 the glycan structures of cell surface proteins 195:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 3 51 theme gene 491:494 arg1 expression 496:505 the gene expression 487:505 the gene expression of various molecules including glycosyltransferases 487:557 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 8 52 theme reduced 1568:1574 arg1 expression 1576:1585 the reduced expression 1564:1585 the reduced expression of the β4GalT1 gene 1564:1605 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 4 53 theme A549 690:693 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 10 54 theme cells 1882:1886 arg1 properties 1863:1872 the malignant properties 1849:1872 the malignant properties of A549 cells 1849:1886 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 0 55 with Line 106:109 arg1 β4-Galactosylation 126:143 Decreased β4-Galactosylation 116:143 Decreased β4-Galactosylation of Highly Branched N-Glycans 116:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 7 56 theme V. 1258:1259 arg1 Mutations 1261:1269 N-acetylglucosaminyltransferase V. Mutations 1226:1269 N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter 1226:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 5 57 theme RNA 822:824 arg1 interference 826:837 RNA interference 822:837 RNA interference 822:837 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 7 58 theme Sp1 1182:1184 arg1 downregulation 1164:1177 downregulation 1164:1177 downregulation of Sp1 1164:1184 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 6 59 theme Lectin 840:845 arg1 blotting 847:854 Lectin blotting 840:854 Lectin blotting 840:854 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 10 60 theme N-glycans 1948:1956 arg1 β4-galactosylation 1910:1927 the decreased β4-galactosylation 1896:1927 the decreased β4-galactosylation of highly branched N-glycans 1896:1956 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 4 61 theme lung 701:704 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 1 62 theme cell 220:223 arg1 proteins 233:240 cell surface proteins 220:240 cell surface proteins 220:240 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 8 63 theme β4GalT1 1594:1600 arg1 gene 1602:1605 the β4GalT1 gene 1590:1605 the β4GalT1 gene 1590:1605 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 4 64 theme cell 713:716 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 7 65 theme gene 1311:1314 arg1 promoter 1316:1323 the β4GalT1 gene promoter 1299:1323 the β4GalT1 gene promoter 1299:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 5 66 theme stable 741:746 arg1 clone 748:752 a stable clone 739:752 a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference 739:837 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 7 67 theme β4GalT 1128:1133 arg1 level 1093:1097 the gene expression level 1073:1097 the gene expression level of β4-galactosyltransferase (β4GalT) 1 1073:1136 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 4 68 gly N-glycosylation 629:643 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 4 68 gly N-glycosylation 629:643 arg1 glycoproteins 648:660 glycoproteins 648:660 glycoproteins 648:660 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 28529241 9 69 from activity 1735:1742 arg1 suppression 1660:1670 the suppression 1656:1670 the suppression of the anchorage-independent growth in soft agar and migratory activity 1656:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 10 70 theme malignant 1853:1861 arg1 properties 1863:1872 the malignant properties 1849:1872 the malignant properties of A549 cells 1849:1886 The present study demonstrates that downregulation of Sp1 suppresses the malignant properties of A549 cells through the decreased β4-galactosylation of highly branched N-glycans. 28529241 5 71 theme control 805:811 arg1 clone 813:817 a control clone 803:817 a control clone 803:817 We established a stable clone whose Sp1-expression level was reduced to 50% of a control clone by RNA interference. 28529241 7 72 theme expression 1082:1091 arg1 level 1093:1097 the gene expression level 1073:1097 the gene expression level of β4-galactosyltransferase (β4GalT) 1 1073:1136 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 73 theme promoter 1316:1323 arg1 promoter 1316:1323 the β4GalT1 gene promoter 1299:1323 the β4GalT1 gene promoter 1299:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 73 theme promoter 1316:1323 arg1 sites 1290:1294 the Sp1-binding sites 1274:1294 the Sp1-binding sites of the β4GalT1 gene promoter 1274:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 0 74 theme Lung 89:92 arg1 Line 106:109 A549 Human Lung Cancer Cell Line 78:109 A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 78:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 1 75 theme glycosyltransferases 347:366 arg1 levels 337:342 the altered expression levels 314:342 the altered expression levels of glycosyltransferases 314:366 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 7 76 from Mutations 1261:1269 arg1 promoter 1316:1323 the β4GalT1 gene promoter 1299:1323 the β4GalT1 gene promoter 1299:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 76 from Mutations 1261:1269 arg1 sites 1290:1294 the Sp1-binding sites 1274:1294 the Sp1-binding sites of the β4GalT1 gene promoter 1274:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 0 77 theme Cell 101:104 arg1 Line 106:109 A549 Human Lung Cancer Cell Line 78:109 A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans 78:172 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 9 78 theme anchorage-independent 1679:1699 arg1 growth 1701:1706 the anchorage-independent growth 1675:1706 the anchorage-independent growth in soft agar and migratory activity 1675:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 9 79 from growth 1701:1706 arg1 agar 1716:1719 soft agar 1711:1719 soft agar 1711:1719 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 9 79 from growth 1701:1706 arg1 activity 1735:1742 migratory activity 1725:1742 migratory activity 1725:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 2 80 theme malignant 414:422 arg1 properties 424:433 the malignant properties 410:433 the malignant properties of cancer cells 410:449 Such changes are closely associated with the malignant properties of cancer cells. 28529241 7 81 theme decreased 1019:1027 arg1 β4-galactosylation 1029:1046 decreased β4-galactosylation 1019:1046 decreased β4-galactosylation of N-glycans 1019:1059 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 3 82 theme factor 466:471 arg1 Sp1 473:475 Transcription factor Sp1 452:475 Transcription factor Sp1 452:475 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 0 83 theme Factor 32:37 arg1 Sp1 39:41 Transcription Factor Sp1 18:41 Transcription Factor Sp1 18:41 Downregulation of Transcription Factor Sp1 Suppresses Malignant Properties of A549 Human Lung Cancer Cell Line with Decreased β4-Galactosylation of Highly Branched N-Glycans. 28529241 7 84 theme mechanism 1005:1013 arg1 analysis 982:989 The analysis 978:989 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans 978:1059 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 9 85 theme migratory 1725:1733 arg1 activity 1735:1742 migratory activity 1725:1742 migratory activity 1725:1742 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 7 86 from changes 1194:1200 arg1 those 1205:1209 those 1205:1209 those 1205:1209 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 8 87 theme branched 1502:1509 arg1 N-glycans 1511:1519 highly branched N-glycans 1495:1519 highly branched N-glycans 1495:1519 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 2 88 theme cells 445:449 arg1 properties 424:433 the malignant properties 410:433 the malignant properties of cancer cells 410:449 Such changes are closely associated with the malignant properties of cancer cells. 28529241 7 89 theme β4GalT1 1303:1309 arg1 promoter 1316:1323 the β4GalT1 gene promoter 1299:1323 the β4GalT1 gene promoter 1299:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 7 90 theme N-acetylglucosaminyltransferase 1226:1256 arg1 Mutations 1261:1269 N-acetylglucosaminyltransferase V. Mutations 1226:1269 N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter 1226:1323 The analysis of underlying mechanism for decreased β4-galactosylation of N-glycans showed that the gene expression level of β4-galactosyltransferase (β4GalT) 1 decreases dramatically by downregulation of Sp1 without changes in those of β4GalT2 and N-acetylglucosaminyltransferase V. Mutations in the Sp1-binding sites of the β4GalT1 gene promoter showed that the promoter activity decreases significantly, indicating that the gene expression is regulated by Sp1. 28529241 1 91 theme altered 318:324 arg1 levels 337:342 the altered expression levels 314:342 the altered expression levels of glycosyltransferases 314:366 Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. 28529241 6 92 theme branched 903:910 arg1 N-glycans 912:920 highly branched N-glycans 896:920 highly branched N-glycans 896:920 Lectin blotting revealed that the β4-galactosylation of highly branched N-glycans decreases mainly in cell adhesion molecule, E-cadherin. 28529241 9 93 theme control 1765:1771 arg1 cells 1773:1777 the control cells 1761:1777 the control cells 1761:1777 Furthermore, the Sp1-downregulated cells showed the suppression of the anchorage-independent growth in soft agar and migratory activity when compared to the control cells. 28529241 8 94 theme Sp1 1552:1554 arg1 downregulation 1534:1547 downregulation 1534:1547 downregulation of Sp1 1534:1554 These results indicate that the β4-galactosylation of highly branched N-glycans decreases by downregulation of Sp1 through the reduced expression of the β4GalT1 gene. 28529241 3 95 theme molecules 518:526 arg1 expression 496:505 the gene expression 487:505 the gene expression of various molecules including glycosyltransferases 487:557 Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. 28529241 4 96 theme human 695:699 arg1 line 718:721 A549 human lung cancer cell line 690:721 A549 human lung cancer cell line 690:721 Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung cancer cell line. 26657071 0 0 theme Carboxylesterase 81:96 arg1 Activity 32:39 Activity 32:39 Activity 32:39 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 0 0 theme Carboxylesterase 81:96 arg1 Structure 18:26 Structure 18:26 Structure 18:26 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 2 1 theme human 461:465 arg1 kidney 477:482 human embryonic kidney 461:482 human embryonic kidney cells 461:488 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 5 2 theme Crystal 772:778 arg1 structures 780:789 Crystal structures 772:789 Crystal structures of hCES1 772:798 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 1 3 theme key 141:143 arg1 enzyme 162:167 the key liver microsomal enzyme 137:167 the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs 137:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 3 theme key 141:143 arg1 Carboxylesterase 107:122 Human Carboxylesterase 1 101:124 Human Carboxylesterase 1 (hCES1) 101:132 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 4 theme drugs 240:244 arg1 variety 220:226 a variety 218:226 a variety of clinical drugs 218:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 4 theme drugs 240:244 arg1 drugs 240:244 clinical drugs 231:244 clinical drugs 231:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 0 5 theme Human 75:79 arg1 Carboxylesterase 81:96 Aglycosylated Human Carboxylesterase 1 61:98 Aglycosylated Human Carboxylesterase 1 61:98 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 1 6 theme liver 145:149 arg1 enzyme 162:167 the key liver microsomal enzyme 137:167 the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs 137:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 6 theme liver 145:149 arg1 Carboxylesterase 107:122 Human Carboxylesterase 1 101:124 Human Carboxylesterase 1 (hCES1) 101:132 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 7 7 theme un-liganded 1187:1197 arg1 enzymes 1199:1205 these un-liganded enzymes 1181:1205 these un-liganded enzymes 1181:1205 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 7 8 theme sites 1172:1176 arg1 Comparison 1147:1156 Comparison 1147:1156 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes 1147:1251 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 5 9 theme high 930:933 arg1 resolutions 935:945 high resolutions 930:945 high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively) 930:987 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 4 10 gly glycosylated 662:673 arg1 hCES1 675:679 glycosylated hCES1 662:679 glycosylated hCES1 662:679 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 4 11 gly aglycosylated 608:620 arg1 active 650:655 active 650:655 active 650:655 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 4 11 gly aglycosylated 608:620 arg1 enzyme 622:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 0 12 gly Aglycosylated 61:73 arg1 Carboxylesterase 81:96 Aglycosylated Human Carboxylesterase 1 61:98 Aglycosylated Human Carboxylesterase 1 61:98 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 7 13 theme hCES1-ligand 1230:1241 arg1 complexes 1243:1251 hCES1-ligand complexes 1230:1251 hCES1-ligand complexes 1230:1251 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 5 14 theme ligand 907:912 arg1 absence 892:898 the absence 888:898 the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively) 888:987 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 6 15 theme conformational 1046:1059 arg1 differences 1061:1071 only minor conformational differences 1035:1071 only minor conformational differences 1035:1071 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 6 16 theme Cα 1133:1134 arg1 positions 1136:1144 all Cα positions 1129:1144 all Cα positions 1129:1144 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 2 17 theme glycosylated 356:367 arg1 hCES1 387:391 authentically glycosylated and aglycosylated hCES1 342:391 authentically glycosylated and aglycosylated hCES1 342:391 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 1 18 theme microsomal 151:160 arg1 enzyme 162:167 the key liver microsomal enzyme 137:167 the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs 137:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 18 theme microsomal 151:160 arg1 Carboxylesterase 107:122 Human Carboxylesterase 1 101:124 Human Carboxylesterase 1 (hCES1) 101:132 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 2 19 gly aglycosylated 373:385 arg1 hCES1 387:391 authentically glycosylated and aglycosylated hCES1 342:391 authentically glycosylated and aglycosylated hCES1 342:391 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 6 20 theme minor 1040:1044 arg1 differences 1061:1071 only minor conformational differences 1035:1071 only minor conformational differences 1035:1071 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 5 21 theme inactive 816:823 arg1 mutant 825:830 a catalytically inactive mutant 800:830 a catalytically inactive mutant (S221A) 800:838 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 5 21 theme inactive 816:823 arg1 S221A 833:837 S221A 833:837 S221A 833:837 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 5 22 gly aglycosylated 848:860 arg1 enzyme 862:867 the aglycosylated enzyme 844:867 the aglycosylated enzyme 844:867 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 2 23 theme kidney 477:482 arg1 cells 484:488 human embryonic kidney cells 461:488 human embryonic kidney cells 461:488 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 2 24 dep structure 304:312 arg1 the 300:302 the 300:302 the 300:302 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 1 25 theme responsible 169:179 arg1 enzyme 162:167 the key liver microsomal enzyme 137:167 the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs 137:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 25 theme responsible 169:179 arg1 Carboxylesterase 107:122 Human Carboxylesterase 1 101:124 Human Carboxylesterase 1 (hCES1) 101:132 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 2 26 gly glycosylated 356:367 arg1 hCES1 387:391 authentically glycosylated and aglycosylated hCES1 342:391 authentically glycosylated and aglycosylated hCES1 342:391 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 2 27 theme glycan 290:295 arg1 role 262:265 the role 258:265 the role of the single N-linked glycan on the structure and activity of the enzyme 258:339 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 3 28 from trimeric 539:546 arg1 solution 551:558 solution 551:558 solution 551:558 Purified enzymes were shown to be predominantly trimeric in solution by analytical ultracentrifugation. 26657071 0 29 theme Structure 18:26 arg1 Comparison 0:9 Comparison 0:9 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 0:99 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 1 30 theme Human 101:105 arg1 hCES1 127:131 hCES1 127:131 hCES1 127:131 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 30 theme Human 101:105 arg1 enzyme 162:167 the key liver microsomal enzyme 137:167 the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs 137:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 30 theme Human 101:105 arg1 Carboxylesterase 107:122 Human Carboxylesterase 1 101:124 Human Carboxylesterase 1 (hCES1) 101:132 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 6 31 theme structures 1017:1026 arg1 Superposition 990:1002 Superposition 990:1002 Superposition of all three structures 990:1026 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 6 32 theme Å 1122:1122 arg1 deviations 1097:1106 a root mean square deviations 1078:1106 a root mean square deviations of around 0.5 Å 1078:1122 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 5 33 dep structures 780:789 arg1 mutant 825:830 a catalytically inactive mutant 800:830 a catalytically inactive mutant (S221A) 800:838 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 5 33 dep structures 780:789 arg1 S221A 833:837 S221A 833:837 S221A 833:837 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 2 34 theme single 274:279 arg1 glycan 290:295 the single N-linked glycan 270:295 the single N-linked glycan 270:295 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 6 35 theme root 1080:1083 arg1 deviations 1097:1106 a root mean square deviations 1078:1106 a root mean square deviations of around 0.5 Å 1078:1122 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 0 36 theme Activity 32:39 arg1 Comparison 0:9 Comparison 0:9 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 0:99 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 4 37 theme glycosylated 662:673 arg1 hCES1 675:679 glycosylated hCES1 662:679 glycosylated hCES1 662:679 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 7 38 with Comparison 1147:1156 arg1 structures 1216:1225 the structures 1212:1225 the structures of hCES1-ligand complexes 1212:1251 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 7 39 theme triad 1294:1298 arg1 side-chains 1265:1275 side-chains 1265:1275 side-chains of the catalytic triad 1265:1298 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 2 40 theme aglycosylated 373:385 arg1 hCES1 387:391 authentically glycosylated and aglycosylated hCES1 342:391 authentically glycosylated and aglycosylated hCES1 342:391 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 2 41 link N-linked 281:288 arg1 glycan 290:295 the single N-linked glycan 270:295 the single N-linked glycan 270:295 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 5 42 theme hCES1 794:798 arg1 enzyme 862:867 the aglycosylated enzyme 844:867 the aglycosylated enzyme 844:867 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 5 42 theme hCES1 794:798 arg1 structures 780:789 Crystal structures 772:789 Crystal structures of hCES1 772:798 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 4 43 theme positive 748:755 arg1 cooperativity 757:769 positive cooperativity 748:769 positive cooperativity 748:769 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 0 44 theme Glycosylated 44:55 arg1 Activity 32:39 Activity 32:39 Activity 32:39 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 0 44 theme Glycosylated 44:55 arg1 Structure 18:26 Structure 18:26 Structure 18:26 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 8 45 dep structure 1445:1453 arg1 the 1441:1443 the 1441:1443 the 1441:1443 Overall the results indicate that preventing N-glycosylation of hCES1 does not significantly affect the structure or activity of the enzyme. 26657071 7 46 theme active 1165:1170 arg1 enzymes 1199:1205 these un-liganded enzymes 1181:1205 these un-liganded enzymes 1181:1205 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 7 46 theme active 1165:1170 arg1 sites 1172:1176 the active sites 1161:1176 the active sites of these un-liganded enzymes 1161:1205 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 6 47 theme square 1090:1095 arg1 deviations 1097:1106 a root mean square deviations 1078:1106 a root mean square deviations of around 0.5 Å 1078:1122 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 4 48 theme kinetics 704:711 arg1 analysis 685:692 analysis 685:692 analysis of enzyme kinetics 685:711 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 5 49 dep resolutions 935:945 arg1 Å 972:972 1.48 Å and 2.01 Å 956:972 1.48 Å and 2.01 Å 956:972 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 5 49 dep resolutions 935:945 arg1 Å 953:953 1.86 Å 948:953 1.86 Å 948:953 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 3 50 theme analytical 563:572 arg1 ultracentrifugation 574:592 analytical ultracentrifugation 563:592 analytical ultracentrifugation 563:592 Purified enzymes were shown to be predominantly trimeric in solution by analytical ultracentrifugation. 26657071 7 51 theme complexes 1243:1251 arg1 structures 1216:1225 the structures 1212:1225 the structures of hCES1-ligand complexes 1212:1251 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 4 52 theme enzyme 697:702 arg1 kinetics 704:711 enzyme kinetics 697:711 enzyme kinetics 697:711 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 2 53 from role 262:265 arg1 activity 318:325 activity 318:325 activity 318:325 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 2 53 from role 262:265 arg1 structure 304:312 structure 304:312 structure 304:312 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 7 54 theme substrate 1322:1330 arg1 binding 1332:1338 substrate binding 1322:1338 substrate binding 1322:1338 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 0 55 dep Structure 18:26 arg1 the 14:16 the 14:16 the 14:16 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 5 56 theme substrate 917:925 arg1 absence 892:898 the absence 888:898 the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively) 888:987 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 2 57 theme N-linked 281:288 arg1 glycan 290:295 the single N-linked glycan 270:295 the single N-linked glycan 270:295 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 4 58 theme purified 599:606 arg1 active 650:655 active 650:655 active 650:655 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 4 58 theme purified 599:606 arg1 enzyme 622:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 8 59 theme enzyme 1474:1479 arg1 structure 1445:1453 structure 1445:1453 structure 1445:1453 Overall the results indicate that preventing N-glycosylation of hCES1 does not significantly affect the structure or activity of the enzyme. 26657071 8 59 theme enzyme 1474:1479 arg1 activity 1458:1465 activity 1458:1465 activity 1458:1465 Overall the results indicate that preventing N-glycosylation of hCES1 does not significantly affect the structure or activity of the enzyme. 26657071 8 60 theme hCES1 1405:1409 arg1 N-glycosylation 1386:1400 N-glycosylation 1386:1400 N-glycosylation of hCES1 1386:1409 Overall the results indicate that preventing N-glycosylation of hCES1 does not significantly affect the structure or activity of the enzyme. 26657071 6 61 theme mean 1085:1088 arg1 deviations 1097:1106 a root mean square deviations 1078:1106 a root mean square deviations of around 0.5 Å 1078:1122 Superposition of all three structures showed only minor conformational differences with a root mean square deviations of around 0.5 Å over all Cα positions. 26657071 7 62 theme catalytic 1284:1292 arg1 triad 1294:1298 the catalytic triad 1280:1298 the catalytic triad 1280:1298 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 1 63 theme variety 220:226 arg1 metabolism 204:213 metabolism 204:213 metabolism 204:213 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 1 63 theme variety 220:226 arg1 detoxification 185:198 detoxification 185:198 detoxification 185:198 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 26657071 0 64 theme Aglycosylated 61:73 arg1 Carboxylesterase 81:96 Aglycosylated Human Carboxylesterase 1 61:98 Aglycosylated Human Carboxylesterase 1 61:98 Comparison of the Structure and Activity of Glycosylated and Aglycosylated Human Carboxylesterase 1. 26657071 5 65 theme aglycosylated 848:860 arg1 enzyme 862:867 the aglycosylated enzyme 844:867 the aglycosylated enzyme 844:867 Crystal structures of hCES1 a catalytically inactive mutant (S221A) and the aglycosylated enzyme were determined in the absence of any ligand or substrate to high resolutions (1.86 Å, 1.48 Å and 2.01 Å, respectively). 26657071 3 66 theme Purified 491:498 arg1 enzymes 500:506 Purified enzymes 491:506 Purified enzymes 491:506 Purified enzymes were shown to be predominantly trimeric in solution by analytical ultracentrifugation. 26657071 3 67 from solution 551:558 arg1 trimeric 539:546 trimeric 539:546 trimeric 539:546 Purified enzymes were shown to be predominantly trimeric in solution by analytical ultracentrifugation. 26657071 8 68 gly N-glycosylation 1386:1400 arg1 hCES1 1405:1409 hCES1 1405:1409 hCES1 1405:1409 Overall the results indicate that preventing N-glycosylation of hCES1 does not significantly affect the structure or activity of the enzyme. 26657071 2 69 theme embryonic 467:475 arg1 kidney 477:482 human embryonic kidney 461:482 human embryonic kidney cells 461:488 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 4 70 theme aglycosylated 608:620 arg1 active 650:655 active 650:655 active 650:655 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 4 70 theme aglycosylated 608:620 arg1 enzyme 622:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme 595:627 The purified aglycosylated enzyme was found to be more active than glycosylated hCES1 and analysis of enzyme kinetics revealed that both enzymes exhibit positive cooperativity. 26657071 2 71 theme enzyme 334:339 arg1 activity 318:325 activity 318:325 activity 318:325 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 2 71 theme enzyme 334:339 arg1 structure 304:312 structure 304:312 structure 304:312 To analyse the role of the single N-linked glycan on the structure and activity of the enzyme, authentically glycosylated and aglycosylated hCES1, generated by mutating asparagine 79 to glutamine, were produced in human embryonic kidney cells. 26657071 7 72 theme enzymes 1199:1205 arg1 enzymes 1199:1205 these un-liganded enzymes 1181:1205 these un-liganded enzymes 1181:1205 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 7 72 theme enzymes 1199:1205 arg1 sites 1172:1176 the active sites 1161:1176 the active sites of these un-liganded enzymes 1161:1205 Comparison of the active sites of these un-liganded enzymes with the structures of hCES1-ligand complexes showed that side-chains of the catalytic triad were pre-disposed for substrate binding. 26657071 1 73 theme clinical 231:238 arg1 drugs 240:244 clinical drugs 231:244 clinical drugs 231:244 Human Carboxylesterase 1 (hCES1) is the key liver microsomal enzyme responsible for detoxification and metabolism of a variety of clinical drugs. 28696719 6 0 theme protein 760:766 arg1 chain 777:781 the protein backbone chain 756:781 the protein backbone chain 756:781 N-Glycosite occupancy varied along the protein backbone chain. 28696719 1 1 theme essential 142:150 arg1 roles 152:156 essential roles 142:156 essential roles 142:156 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 4 2 theme O-glycoforms 612:623 arg1 characterization 585:600 simultaneous characterization 572:600 simultaneous characterization of N- and O-glycoforms 572:623 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 0 3 theme Mass 76:79 arg1 Spectrometry 81:92 Mass Spectrometry 76:92 Mass Spectrometry 76:92 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 5 4 theme HCD 676:678 arg1 mass 687:690 HCD tandem mass 676:690 HCD tandem mass 676:690 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass was utilized to analyze VWF. 28696719 5 5 theme RP-LC-MS/MS 629:639 arg1 system 641:646 An RP-LC-MS/MS system 626:646 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass 626:690 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass was utilized to analyze VWF. 28696719 9 6 theme high 1126:1129 arg1 mannose 1131:1137 high mannose 1126:1137 high mannose 1126:1137 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 13 7 theme profound 1487:1494 arg1 understanding 1496:1508 a profound understanding 1485:1508 a profound understanding of the biological roles of VWF 1485:1539 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 13 8 theme VWF-based 1581:1589 arg1 therapeutics 1591:1602 VWF-based therapeutics 1581:1602 VWF-based therapeutics 1581:1602 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 13 9 theme therapeutics 1591:1602 arg1 production 1567:1576 the production 1563:1576 the production of VWF-based therapeutics 1563:1602 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 11 10 theme unreported 1267:1276 arg1 N-glycosites 1278:1289 previously unreported N-glycosites 1256:1289 previously unreported N-glycosites within domain D'(TIL'-E') 1256:1315 Noteworthy, previously unreported N-glycosites within domain D'(TIL'-E') showed glycosylation. 28696719 5 11 used utilized 696:703 arg2 system 641:646 An RP-LC-MS/MS system 626:646 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass 626:690 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass was utilized to analyze VWF. 28696719 4 12 theme exploration 492:502 arg1 intention 474:482 the intention 470:482 the intention of this exploration 470:502 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 3 13 gly O-glycosites 422:433 arg2 O-glycosites 422:433 O-glycosites 422:433 O-glycosites 422:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 2 14 theme biological 286:295 arg1 phases 297:302 many biological phases 281:302 many biological phases 281:302 There is ample indication that glycosylation affects many biological phases during the protein life cycle. 28696719 8 15 theme bonds 916:920 arg1 cleavage 893:900 Sequential cleavage 882:900 Sequential cleavage of glycosidic bonds along with Human Database mass matching 882:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 3 16 theme comprehensive 344:356 arg1 characterization 358:373 comprehensive characterization 344:373 comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites 344:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 13 17 theme subtle 1418:1423 arg1 characterization 1425:1440 subtle characterization 1418:1440 subtle characterization of site-specific glycoforms 1418:1468 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 13 18 theme VWF 1537:1539 arg1 roles 1528:1532 the biological roles 1513:1532 the biological roles of VWF 1513:1539 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 2 19 theme many 281:284 arg1 phases 297:302 many biological phases 281:302 many biological phases 281:302 There is ample indication that glycosylation affects many biological phases during the protein life cycle. 28696719 8 20 theme glycosidic 905:914 arg1 bonds 916:920 glycosidic bonds 905:920 glycosidic bonds 905:920 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 12 21 located detected 1393:1400 arg1 2298T 1405:1409 2298T 1405:1409 2298T 1405:1409 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 12 21 located detected 1393:1400 arg2 core 1360:1363 sialylated core 1 and core 2 O-glycans 1349:1386 core 1360:1363 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 12 21 located detected 1393:1400 arg2 O-glycans 1378:1386 sialylated core 1 and core 2 O-glycans 1349:1386 O-glycans 1378:1386 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 11 22 gly N-glycosites 1278:1289 arg2 N-glycosites 1278:1289 previously unreported N-glycosites 1256:1289 previously unreported N-glycosites within domain D'(TIL'-E') 1256:1315 Noteworthy, previously unreported N-glycosites within domain D'(TIL'-E') showed glycosylation. 28696719 4 23 gly occupancy 524:532 arg2 N-glycosites 551:562 all canonical N-glycosites 537:562 all canonical N-glycosites 537:562 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 1 24 theme primary 161:167 arg1 hemostasis 169:178 primary hemostasis 161:178 primary hemostasis 161:178 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 3 25 theme simultaneous 404:415 arg1 N-glycosites 391:402 all probable N-glycosites 378:402 all probable N-glycosites simultaneous with O-glycosites 378:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 3 26 gly N-glycosites 391:402 arg2 N-glycosites 391:402 all probable N-glycosites 378:402 all probable N-glycosites simultaneous with O-glycosites 378:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 8 27 dep along 922:926 arg1 with 928:931 with 928:931 with 928:931 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 2 28 theme life 323:326 arg1 cycle 328:332 the protein life cycle 311:332 the protein life cycle 311:332 There is ample indication that glycosylation affects many biological phases during the protein life cycle. 28696719 9 29 theme ABH 1148:1150 arg1 antigen-terminated 1152:1169 ABH antigen-terminated 1148:1169 ABH antigen-terminated 1148:1169 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 12 30 theme core 1371:1374 arg1 O-glycans 1378:1386 sialylated core 1 and core 2 O-glycans 1349:1386 O-glycans 1378:1386 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 13 31 theme glycoforms 1459:1468 arg1 characterization 1425:1440 subtle characterization 1418:1440 subtle characterization of site-specific glycoforms 1418:1468 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 5 32 theme tandem 680:685 arg1 mass 687:690 HCD tandem mass 676:690 HCD tandem mass 676:690 An RP-LC-MS/MS system functionalized with CID and HCD tandem mass was utilized to analyze VWF. 28696719 9 33 theme tetra-antennary 1093:1107 arg1 N-glycans 1109:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 2 34 theme protein 315:321 arg1 cycle 328:332 the protein life cycle 311:332 the protein life cycle 311:332 There is ample indication that glycosylation affects many biological phases during the protein life cycle. 28696719 6 35 theme N-Glycosite 721:731 arg1 occupancy 733:741 N-Glycosite occupancy 721:741 N-Glycosite occupancy 721:741 N-Glycosite occupancy varied along the protein backbone chain. 28696719 3 36 theme probable 382:389 arg1 N-glycosites 391:402 all probable N-glycosites 378:402 all probable N-glycosites simultaneous with O-glycosites 378:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 4 37 theme simultaneous 572:583 arg1 characterization 585:600 simultaneous characterization 572:600 simultaneous characterization of N- and O-glycoforms 572:623 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 1 38 theme plasma 101:106 arg1 VWF 131:133 VWF 131:133 VWF 131:133 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 1 38 theme plasma 101:106 arg1 Factor 123:128 Human plasma von Willebrand Factor 95:128 Human plasma von Willebrand Factor (VWF) 95:134 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 8 39 theme Human 933:937 arg1 mass 948:951 Human Database mass matching 933:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching 882:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 1 40 theme Human 95:99 arg1 VWF 131:133 VWF 131:133 VWF 131:133 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 1 40 theme Human 95:99 arg1 Factor 123:128 Human plasma von Willebrand Factor 95:128 Human plasma von Willebrand Factor (VWF) 95:134 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 4 41 theme N- 605:606 arg1 characterization 585:600 simultaneous characterization 572:600 simultaneous characterization of N- and O-glycoforms 572:623 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 1 42 theme von 108:110 arg1 VWF 131:133 VWF 131:133 VWF 131:133 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 1 42 theme von 108:110 arg1 Factor 123:128 Human plasma von Willebrand Factor 95:128 Human plasma von Willebrand Factor (VWF) 95:134 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 8 43 theme matching 953:960 arg1 mass 948:951 Human Database mass matching 933:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching 882:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 3 44 theme N-glycosites 391:402 arg1 characterization 358:373 comprehensive characterization 344:373 comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites 344:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 0 45 theme Plasma 44:49 arg1 Factor 66:71 Plasma von Willebrand Factor 44:71 Plasma von Willebrand Factor 44:71 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 1 46 with cooperation 183:193 arg1 factors 219:225 other coagulations factors 200:225 other coagulations factors 200:225 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 12 47 theme sialylated 1349:1358 arg1 core 1360:1363 sialylated core 1 and core 2 O-glycans 1349:1386 core 1360:1363 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 9 48 theme sulfated 1176:1183 arg1 N-glycans 1185:1193 sulfated N-glycans 1176:1193 sulfated N-glycans 1176:1193 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 1 49 theme other 200:204 arg1 factors 219:225 other coagulations factors 200:225 other coagulations factors 200:225 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 4 50 theme canonical 541:549 arg1 N-glycosites 551:562 all canonical N-glycosites 537:562 all canonical N-glycosites 537:562 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 9 51 theme glycoforms 1018:1027 arg1 total 1005:1009 A total 1003:1009 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans 1003:1117 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 1 52 theme coagulations 206:217 arg1 factors 219:225 other coagulations factors 200:225 other coagulations factors 200:225 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 0 53 theme Willebrand 55:64 arg1 Factor 66:71 Plasma von Willebrand Factor 44:71 Plasma von Willebrand Factor 44:71 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 7 54 theme characterized 812:824 arg1 O-glycosites 868:879 O-glycosites 868:879 O-glycosites 868:879 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 7 54 theme characterized 812:824 arg1 N- 862:863 N- 862:863 N- 862:863 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 7 54 theme characterized 812:824 arg1 glycoforms 826:835 181 characterized glycoforms 808:835 181 characterized glycoforms 808:835 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 10 55 theme Many 1196:1199 arg1 glycoforms 1201:1210 Many glycoforms 1196:1210 Many glycoforms 1196:1210 Many glycoforms were common across all N-sites. 28696719 1 56 theme Willebrand 112:121 arg1 VWF 131:133 VWF 131:133 VWF 131:133 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 1 56 theme Willebrand 112:121 arg1 Factor 123:128 Human plasma von Willebrand Factor 95:128 Human plasma von Willebrand Factor (VWF) 95:134 Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. 28696719 0 57 theme von 51:53 arg1 Factor 66:71 Plasma von Willebrand Factor 44:71 Plasma von Willebrand Factor 44:71 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 0 58 gly Glyco-Microheterogeneity 16:39 arg1 Factor 66:71 Plasma von Willebrand Factor 44:71 Plasma von Willebrand Factor 44:71 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 8 59 theme glycoform 981:989 arg1 structures 991:1000 the glycoform structures 977:1000 the glycoform structures 977:1000 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 4 60 theme N-glycosites 551:562 arg1 occupancy 524:532 the occupancy 520:532 the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms 520:623 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 13 61 theme biological 1517:1526 arg1 roles 1528:1532 the biological roles 1513:1532 the biological roles of VWF 1513:1539 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 3 62 with simultaneous 404:415 arg1 O-glycosites 422:433 O-glycosites 422:433 O-glycosites 422:433 However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. 28696719 7 63 theme 257 791:793 arg1 spectra 799:805 257 HCD spectra 791:805 257 HCD spectra 791:805 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 13 64 theme site-specific 1445:1457 arg1 glycoforms 1459:1468 site-specific glycoforms 1445:1468 site-specific glycoforms 1445:1468 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 6 65 theme backbone 768:775 arg1 chain 777:781 the protein backbone chain 756:781 the protein backbone chain 756:781 N-Glycosite occupancy varied along the protein backbone chain. 28696719 13 66 theme roles 1528:1532 arg1 understanding 1496:1508 a profound understanding 1485:1508 a profound understanding of the biological roles of VWF 1485:1539 Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics. 28696719 7 67 theme HCD 795:797 arg1 spectra 799:805 257 HCD spectra 791:805 257 HCD spectra 791:805 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 11 68 dep domain 1298:1303 arg1 TIL'-E 1308:1313 TIL'-E' 1308:1314 TIL'-E' 1308:1314 Noteworthy, previously unreported N-glycosites within domain D'(TIL'-E') showed glycosylation. 28696719 11 68 dep domain 1298:1303 arg1 D' 1305:1306 D' 1305:1306 domain D'(TIL'-E') 1298:1315 Noteworthy, previously unreported N-glycosites within domain D'(TIL'-E') showed glycosylation. 28696719 12 69 gly sialylated 1349:1358 arg1 core 1360:1363 sialylated core 1 and core 2 O-glycans 1349:1386 core 1360:1363 Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. 28696719 8 70 theme Database 939:946 arg1 mass 948:951 Human Database mass matching 933:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching 882:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28696719 9 71 theme tri- 1084:1087 arg1 N-glycans 1109:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 9 72 theme biantennary 1055:1065 arg1 N-glycans 1109:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 biantennary and infrequently tri- and tetra-antennary N-glycans 1055:1117 A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. 28696719 0 73 theme Factor 66:71 arg1 Glyco-Microheterogeneity 16:39 Glyco-Microheterogeneity 16:39 Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry 16:92 An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry. 28696719 7 74 gly O-glycosites 868:879 arg2 O-glycosites 868:879 O-glycosites 868:879 O-glycosites 868:879 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 7 74 gly O-glycosites 868:879 arg2 glycoforms 826:835 181 characterized glycoforms 808:835 181 characterized glycoforms 808:835 Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. 28696719 2 75 theme ample 237:241 arg1 indication 243:252 ample indication that glycosylation affects many biological phases during the protein life cycle 237:332 ample indication that glycosylation affects many biological phases during the protein life cycle 237:332 There is ample indication that glycosylation affects many biological phases during the protein life cycle. 28696719 4 76 gly N-glycosites 551:562 arg2 N-glycosites 551:562 all canonical N-glycosites 537:562 all canonical N-glycosites 537:562 Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. 28696719 8 77 theme Sequential 882:891 arg1 cleavage 893:900 Sequential cleavage 882:900 Sequential cleavage of glycosidic bonds along with Human Database mass matching 882:960 Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. 28258464 6 0 theme Byonic 932:937 arg1 tools 948:952 the Byonic or pFind tools 928:952 the Byonic or pFind tools 928:952 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 4 1 theme N-linked 592:599 arg1 glycans 601:607 high-mannose (Man) and hybrid N-linked glycans 562:607 glycans 601:607 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 9 2 theme unique 1477:1482 arg1 sequences 1492:1500 1011 unique peptide sequences 1472:1500 1011 unique peptide sequences 1472:1500 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 12 3 theme fragment 1859:1866 arg1 ions 1868:1871 fragment ions 1859:1871 fragment ions 1859:1871 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 3 theme fragment 1859:1866 arg1 information 1823:1833 the information 1819:1833 the information of intact glycopeptide 1819:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 10 4 theme glycoforms 1642:1651 arg1 ratios 1618:1623 the occupancy ratios 1604:1623 the occupancy ratios of site-specific glycoforms 1604:1651 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 13 5 theme same 2105:2108 arg1 time 2110:2113 the same time 2101:2113 the same time 2101:2113 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 11 6 theme protein 1738:1744 arg1 glycosylation 1746:1758 site-specific protein glycosylation 1724:1758 site-specific protein glycosylation 1724:1758 These results indicate that our method can be used for characterizing site-specific protein glycosylation in complex samples. 28258464 9 7 theme HepG2 1507:1511 arg1 cells 1513:1517 HepG2 cells 1507:1517 HepG2 cells 1507:1517 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 7 8 from information 1025:1035 arg1 glycopeptide 1078:1089 each glycopeptide 1073:1089 each glycopeptide 1073:1089 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 5 9 theme H-treated 642:650 arg1 glycopeptides 652:664 the Endo H-treated glycopeptides 633:664 the Endo H-treated glycopeptides 633:664 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 2 10 theme intact 343:348 arg1 N-glycopeptides 350:364 intact N-glycopeptides 343:364 intact N-glycopeptides in complex proteome samples 343:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 1 11 theme glycan 224:229 arg1 structures 231:240 the glycan structures 220:240 the glycan structures associated with each site 220:266 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 8 12 theme approach 1257:1264 arg1 effectiveness 1235:1247 the effectiveness 1231:1247 the effectiveness of this approach using RNase B and IgG 1231:1286 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 12 13 theme glycopeptide 1845:1856 arg1 ions 1868:1871 fragment ions 1859:1871 fragment ions 1859:1871 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 13 theme glycopeptide 1845:1856 arg1 digestion 1901:1909 endoglycosidase digestion 1885:1909 endoglycosidase digestion 1885:1909 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 13 theme glycopeptide 1845:1856 arg1 reliability 1916:1926 the reliability 1912:1926 the reliability of the identification 1912:1948 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 13 theme glycopeptide 1845:1856 arg1 information 1823:1833 the information 1819:1833 the information of intact glycopeptide 1819:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 7 14 from indication 1041:1050 arg1 glycopeptide 1078:1089 each glycopeptide 1073:1089 each glycopeptide 1073:1089 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 5 15 theme 18O 713:715 arg1 water 717:721 18O water 713:721 18O water 713:721 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 8 16 theme RNase 1272:1276 arg1 B 1278:1278 RNase B 1272:1278 RNase B 1272:1278 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 13 17 gly glycopeptide 2085:2096 arg2 glycopeptide 2085:2096 each glycopeptide 2080:2096 each glycopeptide 2080:2096 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 5 18 theme PNGase 691:696 arg1 F 698:698 PNGase F 691:698 PNGase F treatment in 18O water 691:721 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 6 19 theme pFind 942:946 arg1 tools 948:952 the Byonic or pFind tools 928:952 the Byonic or pFind tools 928:952 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 8 20 theme glycopeptides 1361:1373 arg1 identification 1343:1356 the identification 1339:1356 the identification of glycopeptides from the HepG2 cell line 1339:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 1 21 gly glycosylation 195:207 arg2 glycosylation 195:207 glycosylation 195:207 glycosylation 195:207 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 1 21 gly glycosylation 195:207 arg2 sites 186:190 the sites 182:190 the sites of glycosylation 182:207 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 13 22 theme response 2058:2065 arg1 signaling 2067:2075 the MS response signaling 2051:2075 the MS response signaling of each glycopeptide 2051:2096 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 0 23 theme intact 81:86 arg1 glycopeptides 88:100 intact glycopeptides 81:100 intact glycopeptides 81:100 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 7 24 theme candidate 1004:1012 arg1 information 1025:1035 candidate glycosites information 1004:1035 candidate glycosites information 1004:1035 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 8 25 theme cell 1390:1393 arg1 line 1395:1398 the HepG2 cell line 1380:1398 the HepG2 cell line 1380:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 13 26 theme glycoforms 2015:2024 arg1 ratios 2036:2041 the site-specific glycoforms occupancy ratios 1997:2041 the site-specific glycoforms occupancy ratios 1997:2041 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 2 27 from N-glycopeptides 350:364 arg1 samples 386:392 complex proteome samples 369:392 complex proteome samples 369:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 10 28 from glycosite 1572:1580 arg1 intensity 1524:1532 The intensity 1520:1532 The intensity of different glycoforms at a specific glycosite 1520:1580 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 4 29 theme Endo 544:547 arg1 H 541:541 endoglycosidase H 525:541 endoglycosidase H (Endo H) 525:550 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 4 29 theme Endo 544:547 arg1 H 549:549 Endo H 544:549 Endo H 544:549 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 8 30 from identification 1343:1356 arg1 line 1395:1398 the HepG2 cell line 1380:1398 the HepG2 cell line 1380:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 0 31 gly glycopeptides 88:100 arg2 glycopeptides 88:100 intact glycopeptides 81:100 intact glycopeptides 81:100 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 10 32 theme different 1537:1545 arg1 glycoforms 1547:1556 different glycoforms 1537:1556 different glycoforms 1537:1556 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 1 33 gly glycoprotein 132:143 arg1 glycoprotein 132:143 glycoprotein structures 132:154 glycoprotein structures 132:154 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 0 34 theme endoglycosidase-assisted 38:61 arg1 identification 63:76 endoglycosidase-assisted identification 38:76 endoglycosidase-assisted identification of intact glycopeptides 38:100 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 1 35 theme Detailed 103:110 arg1 characterization 112:127 Detailed characterization 103:127 Detailed characterization of glycoprotein structures 103:154 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 3 36 theme tryptic 414:420 arg1 glycopeptides 422:434 tryptic glycopeptides 414:434 tryptic glycopeptides 414:434 In the first step, tryptic glycopeptides were enriched using ZIC-HILIC. 28258464 0 37 theme Sequential 0:9 arg1 filtering 24:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 10 38 theme specific 1563:1570 arg1 glycosite 1572:1580 a specific glycosite 1561:1580 a specific glycosite 1561:1580 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 0 39 theme ion 20:22 arg1 filtering 24:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 6 40 theme peptide 885:891 arg1 sequences 893:901 the peptide sequences 881:901 the peptide sequences 881:901 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 1 41 theme structures 145:154 arg1 characterization 112:127 Detailed characterization 103:127 Detailed characterization of glycoprotein structures 103:154 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 7 42 theme intact 1179:1184 arg1 identification 1199:1212 intact glycopeptide identification 1179:1212 intact glycopeptide identification 1179:1212 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 6 43 theme glycan 859:864 arg1 structures 866:875 the glycan structures 855:875 the glycan structures 855:875 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 12 44 theme identification 1935:1948 arg1 reliability 1916:1926 the reliability 1912:1926 the reliability of the identification 1912:1948 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 44 theme identification 1935:1948 arg1 information 1823:1833 the information 1819:1833 the information of intact glycopeptide 1819:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 2 45 theme complex 369:375 arg1 samples 386:392 complex proteome samples 369:392 complex proteome samples 369:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 4 46 link N-linked 592:599 arg1 glycans 601:607 high-mannose (Man) and hybrid N-linked glycans 562:607 glycans 601:607 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 8 47 from line 1395:1398 arg1 glycopeptides 1361:1373 glycopeptides 1361:1373 glycopeptides from the HepG2 cell line 1361:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 8 47 from line 1395:1398 arg1 identification 1343:1356 the identification 1339:1356 the identification of glycopeptides from the HepG2 cell line 1339:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 11 48 theme site-specific 1724:1736 arg1 glycosylation 1746:1758 site-specific protein glycosylation 1724:1758 site-specific protein glycosylation 1724:1758 These results indicate that our method can be used for characterizing site-specific protein glycosylation in complex samples. 28258464 5 49 theme glycopeptides 652:664 arg1 fraction 621:628 a fraction 619:628 a fraction of the Endo H-treated glycopeptides 619:664 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 9 50 theme 1011 1472:1475 arg1 sequences 1492:1500 1011 unique peptide sequences 1472:1500 1011 unique peptide sequences 1472:1500 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 4 51 theme hybrid 585:590 arg1 glycans 601:607 high-mannose (Man) and hybrid N-linked glycans 562:607 glycans 601:607 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 9 52 theme peptide 1484:1490 arg1 sequences 1492:1500 1011 unique peptide sequences 1472:1500 1011 unique peptide sequences 1472:1500 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 2 53 from samples 386:392 arg1 characterization 323:338 characterization 323:338 characterization of intact N-glycopeptides in complex proteome samples 323:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 10 54 theme site-specific 1628:1640 arg1 glycoforms 1642:1651 site-specific glycoforms 1628:1651 site-specific glycoforms 1628:1651 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 11 55 theme complex 1763:1769 arg1 samples 1771:1777 complex samples 1763:1777 complex samples 1763:1777 These results indicate that our method can be used for characterizing site-specific protein glycosylation in complex samples. 28258464 7 56 theme search 1129:1134 arg1 space 1136:1140 the database search space 1116:1140 the database search space 1116:1140 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 2 57 theme analytical 299:308 arg1 strategy 310:317 an analytical strategy 296:317 an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples 296:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 11 58 gly glycosylation 1746:1758 arg1 samples 1771:1777 complex samples 1763:1777 complex samples 1763:1777 These results indicate that our method can be used for characterizing site-specific protein glycosylation in complex samples. 28258464 12 59 theme intact 1838:1843 arg1 glycopeptide 1845:1856 intact glycopeptide 1838:1856 intact glycopeptide 1838:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 10 60 gly glycosite 1572:1580 arg2 glycosite 1572:1580 a specific glycosite 1561:1580 a specific glycosite 1561:1580 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 5 61 gly glycopeptides 652:664 arg2 glycopeptides 652:664 the Endo H-treated glycopeptides 633:664 the Endo H-treated glycopeptides 633:664 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 5 62 theme Endo 637:640 arg1 glycopeptides 652:664 the Endo H-treated glycopeptides 633:664 the Endo H-treated glycopeptides 633:664 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 6 63 theme deglycosylated 793:806 arg1 peptides 808:815 deglycosylated peptides 793:815 deglycosylated peptides 793:815 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 2 64 theme N-glycopeptides 350:364 arg1 characterization 323:338 characterization 323:338 characterization of intact N-glycopeptides in complex proteome samples 323:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 11 65 used used 1700:1703 arg2 method 1686:1691 our method 1682:1691 our method 1682:1691 These results indicate that our method can be used for characterizing site-specific protein glycosylation in complex samples. 28258464 6 66 gly deglycosylated 793:806 arg1 peptides 808:815 deglycosylated peptides 793:815 deglycosylated peptides 793:815 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 7 67 theme database 1120:1127 arg1 space 1136:1140 the database search space 1116:1140 the database search space 1116:1140 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 13 68 theme glycopeptide 2085:2096 arg1 signaling 2067:2075 the MS response signaling 2051:2075 the MS response signaling of each glycopeptide 2051:2096 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 6 69 gly glycopeptides 775:787 arg2 glycopeptides 775:787 The intact glycopeptides 764:787 The intact glycopeptides 764:787 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 8 70 theme sequential 1305:1314 arg1 digestion 1316:1324 sequential digestion 1305:1324 this sequential digestion strategy 1300:1333 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 13 71 theme occupancy 2026:2034 arg1 ratios 2036:2041 the site-specific glycoforms occupancy ratios 1997:2041 the site-specific glycoforms occupancy ratios 1997:2041 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 4 72 theme glycopeptides 494:506 arg1 portion 479:485 a portion 477:485 a portion of the glycopeptides 477:506 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 12 73 gly glycopeptide 1845:1856 arg2 glycopeptide 1845:1856 intact glycopeptide 1838:1856 intact glycopeptide 1838:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 7 74 theme glycoforms 1059:1068 arg1 information 1025:1035 candidate glycosites information 1004:1035 candidate glycosites information 1004:1035 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 7 74 theme glycoforms 1059:1068 arg1 indication 1041:1050 indication 1041:1050 indication of the glycoforms 1041:1068 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 7 75 gly glycopeptide 1078:1089 arg2 glycopeptide 1078:1089 each glycopeptide 1073:1089 each glycopeptide 1073:1089 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 5 76 theme F 698:698 arg1 treatment 700:708 PNGase F treatment 691:708 PNGase F treatment in 18O water 691:721 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 13 77 theme MS 2055:2056 arg1 signaling 2067:2075 the MS response signaling 2051:2075 the MS response signaling of each glycopeptide 2051:2096 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 0 78 theme glycopeptides 88:100 arg1 filtering 24:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 0 78 theme glycopeptides 88:100 arg1 identification 63:76 endoglycosidase-assisted identification 38:76 endoglycosidase-assisted identification of intact glycopeptides 38:100 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 8 79 theme digestion 1316:1324 arg1 strategy 1326:1333 this sequential digestion strategy 1300:1333 this sequential digestion strategy 1300:1333 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 13 80 theme site-specific 2001:2013 arg1 ratios 2036:2041 the site-specific glycoforms occupancy ratios 1997:2041 the site-specific glycoforms occupancy ratios 1997:2041 We quantified the site-specific glycoforms occupancy ratios through the MS response signaling of each glycopeptide at the same time. 28258464 7 81 theme glycosites 1014:1023 arg1 information 1025:1035 candidate glycosites information 1004:1035 candidate glycosites information 1004:1035 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 1 82 theme glycosylation 195:207 arg1 structures 231:240 the glycan structures 220:240 the glycan structures associated with each site 220:266 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 1 82 theme glycosylation 195:207 arg1 glycosylation 195:207 glycosylation 195:207 glycosylation 195:207 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 1 82 theme glycosylation 195:207 arg1 determining 165:175 determining 165:175 determining 165:175 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 1 82 theme glycosylation 195:207 arg1 sites 186:190 the sites 182:190 the sites of glycosylation 182:207 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 2 83 from characterization 323:338 arg1 samples 386:392 complex proteome samples 369:392 complex proteome samples 369:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 8 84 theme HepG2 1384:1388 arg1 line 1395:1398 the HepG2 cell line 1380:1398 the HepG2 cell line 1380:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 12 85 dep ions 1868:1871 arg1 filters 1873:1879 filters 1873:1879 filters 1873:1879 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 8 86 gly glycopeptides 1361:1373 arg1 line 1395:1398 the HepG2 cell line 1380:1398 the HepG2 cell line 1380:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 8 86 gly glycopeptides 1361:1373 arg2 glycopeptides 1361:1373 glycopeptides 1361:1373 glycopeptides from the HepG2 cell line 1361:1398 We demonstrated the effectiveness of this approach using RNase B and IgG and applied this sequential digestion strategy for the identification of glycopeptides from the HepG2 cell line. 28258464 12 87 theme Graphical 1780:1788 arg1 abstract 1790:1797 Graphical abstract 1780:1797 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification 1780:1948 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 3 88 theme first 402:406 arg1 step 408:411 the first step 398:411 the first step 398:411 In the first step, tryptic glycopeptides were enriched using ZIC-HILIC. 28258464 5 89 theme remaining 737:745 arg1 glycans 755:761 the remaining complex glycans 733:761 the remaining complex glycans 733:761 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 3 90 gly glycopeptides 422:434 arg2 glycopeptides 422:434 tryptic glycopeptides 414:434 tryptic glycopeptides 414:434 In the first step, tryptic glycopeptides were enriched using ZIC-HILIC. 28258464 4 91 theme endoglycosidase 525:539 arg1 H 541:541 endoglycosidase H 525:541 endoglycosidase H (Endo H) 525:550 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 4 91 theme endoglycosidase 525:539 arg1 H 549:549 Endo H 544:549 Endo H 544:549 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 5 92 from treatment 700:708 arg1 water 717:721 18O water 713:721 18O water 713:721 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 0 93 theme fragment 11:18 arg1 filtering 24:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering 0:32 Sequential fragment ion filtering and endoglycosidase-assisted identification of intact glycopeptides. 28258464 7 94 theme Sequential 955:964 arg1 digestion 966:974 Sequential digestion 955:974 Sequential digestion by endoglycosidase 955:993 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 1 95 theme glycoprotein 132:143 arg1 structures 145:154 glycoprotein structures 132:154 glycoprotein structures 132:154 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 7 96 gly glycosites 1014:1023 arg2 glycosites 1014:1023 candidate glycosites information 1004:1035 candidate glycosites information 1004:1035 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 9 97 gly glycopeptides 1427:1439 arg2 glycopeptides 1427:1439 4514 intact glycopeptides 1415:1439 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells 1415:1517 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 7 98 theme glycopeptide 1186:1197 arg1 identification 1199:1212 intact glycopeptide identification 1179:1212 intact glycopeptide identification 1179:1212 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 4 99 gly glycopeptides 494:506 arg2 glycopeptides 494:506 the glycopeptides 490:506 the glycopeptides 490:506 Secondly, a portion of the glycopeptides was treated with endoglycosidase H (Endo H) to remove high-mannose (Man) and hybrid N-linked glycans. 28258464 10 100 theme glycoforms 1547:1556 arg1 intensity 1524:1532 The intensity 1520:1532 The intensity of different glycoforms at a specific glycosite 1520:1580 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 28258464 7 101 gly glycopeptide 1186:1197 arg2 glycopeptide 1186:1197 intact glycopeptide identification 1179:1212 intact glycopeptide identification 1179:1212 Sequential digestion by endoglycosidase provided candidate glycosites information and indication of the glycoforms on each glycopeptide, thus helping to confine the database search space and improve the confidence regarding intact glycopeptide identification. 28258464 6 102 theme intact 768:773 arg1 glycopeptides 775:787 The intact glycopeptides 764:787 The intact glycopeptides 764:787 The intact glycopeptides and deglycosylated peptides were analyzed by nano-RPLC-MS/MS, and the glycan structures and the peptide sequences were identified by using the Byonic or pFind tools. 28258464 5 103 theme complex 747:753 arg1 glycans 755:761 the remaining complex glycans 733:761 the remaining complex glycans 733:761 Thirdly, a fraction of the Endo H-treated glycopeptides was further subjected to PNGase F treatment in 18O water to remove the remaining complex glycans. 28258464 9 104 theme intact 1420:1425 arg1 glycopeptides 1427:1439 4514 intact glycopeptides 1415:1439 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells 1415:1517 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 2 105 gly N-glycopeptides 350:364 arg2 N-glycopeptides 350:364 intact N-glycopeptides 343:364 intact N-glycopeptides in complex proteome samples 343:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 9 106 gly glycosites 1457:1466 arg2 glycosites 1457:1466 947 glycosites 1453:1466 947 glycosites 1453:1466 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 2 107 theme proteome 377:384 arg1 samples 386:392 complex proteome samples 369:392 complex proteome samples 369:392 In this work, we developed an analytical strategy for characterization of intact N-glycopeptides in complex proteome samples. 28258464 1 108 dep determining 165:175 arg1 both 177:180 both 177:180 both 177:180 Detailed characterization of glycoprotein structures requires determining both the sites of glycosylation as well as the glycan structures associated with each site. 28258464 9 109 theme 947 1453:1455 arg1 glycosites 1457:1466 947 glycosites 1453:1466 947 glycosites 1453:1466 We identified 4514 intact glycopeptides coming from 947 glycosites and 1011 unique peptide sequences from HepG2 cells. 28258464 12 110 theme endoglycosidase 1885:1899 arg1 digestion 1901:1909 endoglycosidase digestion 1885:1909 endoglycosidase digestion 1885:1909 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 12 110 theme endoglycosidase 1885:1899 arg1 information 1823:1833 the information 1819:1833 the information of intact glycopeptide 1819:1856 Graphical abstract Through integrating the information of intact glycopeptide, fragment ions filters and endoglycosidase digestion, the reliability of the identification could be significantly improved. 28258464 10 111 theme occupancy 1608:1616 arg1 ratios 1618:1623 the occupancy ratios 1604:1623 the occupancy ratios of site-specific glycoforms 1604:1651 The intensity of different glycoforms at a specific glycosite was obtained to reach the occupancy ratios of site-specific glycoforms. 24325898 9 0 theme age 1624:1626 arg1 biomarkers 1610:1619 other biomarkers 1604:1619 other biomarkers of age 1604:1626 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 8 1 with glycosylation 1467:1479 arg1 age 1486:1488 age 1486:1488 age 1486:1488 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 9 2 theme telomere 1633:1640 arg1 lengths 1642:1648 telomere lengths 1633:1648 telomere lengths 1633:1648 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 1 3 theme structural 73:82 arg1 details 84:90 Fine structural details 68:90 Fine structural details of glycans attached to the conserved N-glycosylation site 68:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 10 4 theme biological 1757:1766 arg1 age 1768:1770 biological age 1757:1770 biological age 1757:1770 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 3 5 theme other 663:667 arg1 markers 669:675 other markers 663:675 other markers of biological age like telomere lengths 663:715 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 6 theme telomere 700:707 arg1 lengths 709:715 telomere lengths 700:715 telomere lengths 700:715 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 2 7 theme IgG 302:304 arg1 glycosylation 306:318 IgG glycosylation 302:318 IgG glycosylation 302:318 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 5 8 theme IgG 846:848 arg1 glycosylation 850:862 IgG glycosylation 846:862 IgG glycosylation 846:862 Thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 2 9 from populations 360:370 arg1 individuals 329:339 5,117 individuals 323:339 5,117 individuals from four European populations 323:370 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 3 10 theme age 691:693 arg1 markers 669:675 other markers 663:675 other markers of biological age like telomere lengths 663:715 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 1 11 theme immunoglobulin 203:216 arg1 IgG 221:223 IgG 221:223 IgG 221:223 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 1 11 theme immunoglobulin 203:216 arg1 G 218:218 immunoglobulin G 203:218 individual immunoglobulin G (IgG) molecules 192:234 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 8 12 theme complex 1444:1450 arg1 changes 1452:1458 very extensive and complex changes 1425:1458 very extensive and complex changes in IgG glycosylation with age 1425:1488 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 11 13 theme diverse 1974:1980 arg1 treatment 2020:2028 treatment 2020:2028 treatment 2020:2028 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 11 13 theme diverse 1974:1980 arg1 forensics 2034:2042 forensics 2034:2042 forensics 2034:2042 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 11 13 theme diverse 1974:1980 arg1 prevention 2005:2014 disease prevention 1997:2014 disease prevention 1997:2014 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 11 13 theme diverse 1974:1980 arg1 fields 1982:1987 diverse fields 1974:1987 diverse fields such as disease prevention and treatment, or forensics 1974:2042 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 1 14 gly N-glycosylation 129:143 arg2 site 145:148 the conserved N-glycosylation site 115:148 the conserved N-glycosylation site 115:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 9 15 theme combined 1495:1502 arg1 index 1504:1508 The combined index 1491:1508 The combined index composed of only three glycans 1491:1539 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 9 16 theme variance 1564:1571 arg1 variance 1564:1571 variance 1564:1571 variance 1564:1571 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 9 16 theme variance 1564:1571 arg1 % 1559:1559 up to 58% 1551:1559 up to 58% of variance 1551:1571 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 11 17 theme disease 1997:2003 arg1 prevention 2005:2014 disease prevention 1997:2014 disease prevention 1997:2014 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 1 18 theme conserved 119:127 arg1 site 145:148 the conserved N-glycosylation site 115:148 the conserved N-glycosylation site 115:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 3 19 theme Several 454:460 arg1 FA2G2 491:495 FA2G2 491:495 FA2G2 491:495 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 19 theme Several 454:460 arg1 glycans 466:472 Several IgG glycans 454:472 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) 454:508 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 19 theme Several 454:460 arg1 FA2B 485:488 FA2B 485:488 FA2B 485:488 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 19 theme Several 454:460 arg1 FA2BG2 502:507 FA2BG2 502:507 FA2BG2 502:507 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 0 20 theme biological 51:60 arg1 ages 62:65 chronological and biological ages 33:65 chronological and biological ages 33:65 Glycans are a novel biomarker of chronological and biological ages. 24325898 7 21 theme key 1194:1196 arg1 Glycosylation 1173:1185 Glycosylation 1173:1185 Glycosylation 1173:1185 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 7 21 theme key 1194:1196 arg1 mechanism 1216:1224 the key posttranslational mechanism 1190:1224 the key posttranslational mechanism that regulates function of immunoglobulins 1190:1267 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 4 22 from variance 732:739 arg1 glycans 750:756 these glycans 744:756 these glycans 744:756 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age. 24325898 6 23 theme IgG 972:974 arg1 glycans 976:982 IgG glycans 972:982 IgG glycans 972:982 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 2 24 from glycosylation 430:442 arg1 patterns 403:410 very complex patterns 390:410 very complex patterns of changes in IgG glycosylation with age 390:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 10 25 theme remaining 1655:1663 arg1 variance 1665:1672 The remaining variance 1651:1672 The remaining variance in these glycans 1651:1689 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 8 26 theme glycosylation 1346:1358 arg1 study 1333:1337 Our study 1329:1337 Our study of IgG glycosylation in 5,117 individuals from four European populations 1329:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 11 27 theme biological 1900:1909 arg1 aging 1911:1915 human biological aging 1894:1915 human biological aging 1894:1915 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 1 28 theme Fine 68:71 arg1 details 84:90 Fine structural details 68:90 Fine structural details of glycans attached to the conserved N-glycosylation site 68:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 0 29 theme ages 62:65 arg1 Glycans 0:6 Glycans 0:6 Glycans 0:6 Glycans are a novel biomarker of chronological and biological ages. 24325898 0 29 theme ages 62:65 arg1 biomarker 20:28 a novel biomarker 12:28 a novel biomarker of chronological and biological ages 12:65 Glycans are a novel biomarker of chronological and biological ages. 24325898 7 30 theme SIGNIFICANCE 1149:1160 arg1 STATEMENT 1162:1170 SIGNIFICANCE STATEMENT 1149:1170 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 1149:1327 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 2 31 theme IgG 426:428 arg1 glycosylation 430:442 IgG glycosylation 426:442 IgG glycosylation with age 426:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 2 32 theme changes 415:421 arg1 patterns 403:410 very complex patterns 390:410 very complex patterns of changes in IgG glycosylation with age 390:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 1 33 theme glycans 95:101 arg1 details 84:90 Fine structural details 68:90 Fine structural details of glycans attached to the conserved N-glycosylation site 68:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 6 34 theme observed 1017:1024 arg1 changes 1026:1032 the observed changes 1013:1032 the observed changes with age 1013:1041 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 10 35 theme biological 1854:1863 arg1 ages 1865:1868 both chronological and biological ages 1831:1868 both chronological and biological ages 1831:1868 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 8 36 theme European 1391:1398 arg1 populations 1400:1410 four European populations 1386:1410 four European populations 1386:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 7 37 dep STATEMENT 1162:1170 arg1 Glycosylation 1173:1185 Glycosylation 1173:1185 Glycosylation 1173:1185 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 7 37 dep STATEMENT 1162:1170 arg1 mechanism 1216:1224 the key posttranslational mechanism 1190:1224 the key posttranslational mechanism that regulates function of immunoglobulins 1190:1267 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 10 38 theme chronological 1836:1848 arg1 ages 1865:1868 both chronological and biological ages 1831:1868 both chronological and biological ages 1831:1868 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 0 39 theme chronological 33:45 arg1 ages 62:65 chronological and biological ages 33:65 chronological and biological ages 33:65 Glycans are a novel biomarker of chronological and biological ages. 24325898 9 40 theme more 1594:1597 arg1 age 1576:1578 age 1576:1578 age 1576:1578 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 7 41 theme multiple 1275:1282 arg1 repercussions 1293:1305 multiple systemic repercussions 1275:1305 multiple systemic repercussions to the immune system 1275:1326 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 6 42 with changes 1026:1032 arg1 age 1039:1041 age 1039:1041 age 1039:1041 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 9 43 theme other 1604:1608 arg1 biomarkers 1610:1619 other biomarkers 1604:1619 other biomarkers of age 1604:1626 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 2 44 from patterns 403:410 arg1 glycosylation 430:442 IgG glycosylation 426:442 IgG glycosylation with age 426:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 11 45 theme molecular 1923:1931 arg1 profiling 1933:1941 molecular profiling 1923:1941 molecular profiling 1923:1941 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 8 46 from study 1333:1337 arg1 populations 1400:1410 four European populations 1386:1410 four European populations 1386:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 8 46 from study 1333:1337 arg1 individuals 1369:1379 5,117 individuals 1363:1379 5,117 individuals from four European populations 1363:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 1 47 theme molecules 226:234 arg1 function 180:187 function 180:187 function of individual immunoglobulin G (IgG) molecules 180:234 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 10 48 from variance 1665:1672 arg1 glycans 1683:1689 these glycans 1677:1689 these glycans 1677:1689 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 1 49 attach attached 103:110 arg2 glycans 95:101 glycans 95:101 glycans attached to the conserved N-glycosylation site 95:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 1 49 attach attached 103:110 arg1 site 145:148 the conserved N-glycosylation site 115:148 the conserved N-glycosylation site 115:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 2 50 theme European 351:358 arg1 populations 360:370 four European populations 346:370 four European populations 346:370 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 3 51 theme biological 680:689 arg1 age 691:693 biological age 680:693 biological age 680:693 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 8 52 from populations 1400:1410 arg1 study 1333:1337 Our study 1329:1337 Our study of IgG glycosylation in 5,117 individuals from four European populations 1329:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 8 52 from populations 1400:1410 arg1 individuals 1369:1379 5,117 individuals 1363:1379 5,117 individuals from four European populations 1363:1410 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 10 53 theme IgG 1779:1781 arg1 glycosylation 1783:1795 IgG glycosylation 1779:1795 IgG glycosylation 1779:1795 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 6 54 theme important 954:962 arg1 role 964:967 the important role 950:967 the important role of IgG glycans in inflammation 950:998 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 4 55 theme biological 824:833 arg1 age 835:837 biological age 824:837 biological age 824:837 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age. 24325898 1 56 theme individual 192:201 arg1 molecules 226:234 individual immunoglobulin G (IgG) molecules 192:234 individual immunoglobulin G (IgG) molecules 192:234 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 1 57 theme G 218:218 arg1 molecules 226:234 individual immunoglobulin G (IgG) molecules 192:234 individual immunoglobulin G (IgG) molecules 192:234 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 8 58 theme IgG 1463:1465 arg1 glycosylation 1467:1479 IgG glycosylation 1463:1479 IgG glycosylation with age 1463:1488 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 11 59 theme practical 1947:1955 arg1 applications 1957:1968 practical applications 1947:1968 practical applications for diverse fields such as disease prevention and treatment, or forensics 1947:2042 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 9 60 dep 58 1557:1558 arg1 to 1554:1555 to 1554:1555 to 1554:1555 The combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths. 24325898 2 61 with glycosylation 430:442 arg1 age 449:451 age 449:451 age 449:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 7 62 theme posttranslational 1198:1214 arg1 Glycosylation 1173:1185 Glycosylation 1173:1185 Glycosylation 1173:1185 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 7 62 theme posttranslational 1198:1214 arg1 mechanism 1216:1224 the key posttranslational mechanism 1190:1224 the key posttranslational mechanism that regulates function of immunoglobulins 1190:1267 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 3 63 theme glycans 575:581 arg1 combination 548:558 the combination 544:558 the combination of these three glycans 544:581 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 1 64 theme N-glycosylation 129:143 arg1 site 145:148 the conserved N-glycosylation site 115:148 the conserved N-glycosylation site 115:148 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 7 65 with mechanism 1216:1224 arg1 repercussions 1293:1305 multiple systemic repercussions 1275:1305 multiple systemic repercussions to the immune system 1275:1326 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 8 66 from changes 1452:1458 arg1 glycosylation 1467:1479 IgG glycosylation 1463:1479 IgG glycosylation with age 1463:1488 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 6 67 theme glycans 976:982 arg1 role 964:967 the important role 950:967 the important role of IgG glycans in inflammation 950:998 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 11 68 contain has 1943:1945 arg1 ability 1875:1881 The ability to measure human biological aging using molecular profiling 1871:1941 The ability to measure human biological aging using molecular profiling 1871:1941 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 11 68 contain has 1943:1945 arg2 applications 1957:1968 practical applications 1947:1968 practical applications for diverse fields such as disease prevention and treatment, or forensics 1947:2042 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 6 69 from role 964:967 arg1 inflammation 987:998 inflammation 987:998 inflammation 987:998 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 8 70 theme IgG 1342:1344 arg1 glycosylation 1346:1358 IgG glycosylation 1342:1358 IgG glycosylation 1342:1358 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 5 71 theme chronological 903:915 arg1 ages 932:935 both chronological and biological ages 898:935 both chronological and biological ages 898:935 Thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 7 72 theme immune 1314:1319 arg1 system 1321:1326 the immune system 1310:1326 the immune system 1310:1326 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 11 73 theme human 1894:1898 arg1 aging 1911:1915 human biological aging 1894:1915 human biological aging 1894:1915 The ability to measure human biological aging using molecular profiling has practical applications for diverse fields such as disease prevention and treatment, or forensics. 24325898 0 74 theme novel 14:18 arg1 Glycans 0:6 Glycans 0:6 Glycans 0:6 Glycans are a novel biomarker of chronological and biological ages. 24325898 0 74 theme novel 14:18 arg1 biomarker 20:28 a novel biomarker 12:28 a novel biomarker of chronological and biological ages 12:65 Glycans are a novel biomarker of chronological and biological ages. 24325898 1 75 theme systemic 273:280 arg1 level 282:286 the systemic level 269:286 the systemic level 269:286 Fine structural details of glycans attached to the conserved N-glycosylation site significantly not only affect function of individual immunoglobulin G (IgG) molecules but also mediate inflammation at the systemic level. 24325898 5 76 theme biological 921:930 arg1 ages 932:935 both chronological and biological ages 898:935 both chronological and biological ages 898:935 Thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 24325898 3 77 theme IgG 462:464 arg1 FA2G2 491:495 FA2G2 491:495 FA2G2 491:495 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 77 theme IgG 462:464 arg1 glycans 466:472 Several IgG glycans 454:472 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) 454:508 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 77 theme IgG 462:464 arg1 FA2B 485:488 FA2B 485:488 FA2B 485:488 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 77 theme IgG 462:464 arg1 FA2BG2 502:507 FA2BG2 502:507 FA2BG2 502:507 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 7 78 theme systemic 1284:1291 arg1 repercussions 1293:1305 multiple systemic repercussions 1275:1305 multiple systemic repercussions to the immune system 1275:1326 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 2 79 theme complex 395:401 arg1 patterns 403:410 very complex patterns 390:410 very complex patterns of changes in IgG glycosylation with age 390:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 3 80 theme chronological 620:632 arg1 age 634:636 chronological age 620:636 chronological age 620:636 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 4 81 theme physiological 783:795 arg1 parameters 797:806 physiological parameters 783:806 physiological parameters associated with biological age 783:837 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age. 24325898 7 82 theme immunoglobulins 1253:1267 arg1 function 1241:1248 function 1241:1248 function of immunoglobulins 1241:1267 SIGNIFICANCE STATEMENT: Glycosylation is the key posttranslational mechanism that regulates function of immunoglobulins, with multiple systemic repercussions to the immune system. 24325898 2 83 from changes 415:421 arg1 glycosylation 430:442 IgG glycosylation 426:442 IgG glycosylation with age 426:451 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 4 84 theme remaining 722:730 arg1 variance 732:739 The remaining variance 718:739 The remaining variance in these glycans 718:756 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age. 24325898 3 85 theme variance 608:615 arg1 % 603:603 up to 58% 595:603 up to 58% of variance 595:615 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 3 85 theme variance 608:615 arg1 variance 608:615 variance 608:615 variance 608:615 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 6 86 from changes 1065:1071 arg1 glycosylation 1080:1092 IgG glycosylation 1076:1092 IgG glycosylation 1076:1092 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 3 87 dep 58 601:602 arg1 to 598:599 to 598:599 to 598:599 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 6 88 theme IgG 1076:1078 arg1 glycosylation 1080:1092 IgG glycosylation 1076:1092 IgG glycosylation 1076:1092 Considering the important role of IgG glycans in inflammation, and because the observed changes with age promote inflammation, changes in IgG glycosylation also seem to represent a factor contributing to aging. 24325898 2 89 gly glycosylation 306:318 arg1 individuals 329:339 5,117 individuals 323:339 5,117 individuals from four European populations 323:370 By analyzing IgG glycosylation in 5,117 individuals from four European populations, we have revealed very complex patterns of changes in IgG glycosylation with age. 24325898 3 90 theme more 653:656 arg1 age 634:636 chronological age 620:636 chronological age 620:636 Several IgG glycans (including FA2B, FA2G2, and FA2BG2) changed considerably with age and the combination of these three glycans can explain up to 58% of variance in chronological age, significantly more than other markers of biological age like telomere lengths. 24325898 8 91 theme extensive 1430:1438 arg1 changes 1452:1458 very extensive and complex changes 1425:1458 very extensive and complex changes in IgG glycosylation with age 1425:1488 Our study of IgG glycosylation in 5,117 individuals from four European populations has revealed very extensive and complex changes in IgG glycosylation with age. 24325898 10 92 theme physiological 1716:1728 arg1 parameters 1730:1739 physiological parameters 1716:1739 physiological parameters associated with biological age 1716:1770 The remaining variance in these glycans strongly correlated with physiological parameters associated with biological age; thus, IgG glycosylation appears to be closely linked with both chronological and biological ages. 29119347 4 0 theme downstream 1448:1457 arg1 phosphorylation 1476:1490 downstream in vitro mannose phosphorylation 1448:1490 downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1448:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 1 theme N-glycan 210:217 arg1 mutants 230:236 Arabidopsis N-glycan processing mutants 198:236 Arabidopsis N-glycan processing mutants 198:236 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 4 2 dep in 1459:1460 arg1 vitro 1462:1466 vitro 1462:1466 vitro 1462:1466 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 6 3 theme gm1-IDUA 1919:1926 arg1 N-glycans 1906:1914 the high-mannose N-glycans 1889:1914 the high-mannose N-glycans of gm1-IDUA 1889:1926 This may be due to the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA. 29119347 4 4 theme IDUA 1409:1412 arg1 proteins 1414:1421 the two IDUA proteins 1401:1421 the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1401:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 8 5 dep platforms 2164:2172 arg1 create 2177:2182 create 2177:2182 to create enzyme replacement therapeutics for lysosomal storage diseases 2174:2245 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 4 6 theme lysosomal 1234:1242 arg1 enzymes 1244:1250 recombinant lysosomal enzymes 1222:1250 recombinant lysosomal enzymes produced in plants 1222:1269 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 7 theme lysosomal 416:424 arg1 trafficking 426:436 lysosomal trafficking 416:436 lysosomal trafficking 416:436 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 7 8 theme % 2058:2058 arg1 TSP 2060:2062 1.5% TSP 2055:2062 1.5% TSP 2055:2062 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 3 9 theme kinetic 1109:1115 arg1 parameters 1117:1126 their kinetic parameters 1103:1126 their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide 1103:1211 Both forms of IDUA were equivalent with respect to their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide. 29119347 1 10 theme gm1 733:735 arg1 mutant 737:742 the Arabidopsis gm1 mutant 717:742 the Arabidopsis gm1 mutant 717:742 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 4 11 theme soluble 1506:1512 arg1 form 1514:1517 a soluble form 1504:1517 a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1504:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 12 theme therapy 148:154 arg1 development 114:124 development 114:124 development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE 114:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 1 13 theme Functional 452:461 arg1 alpha-L-iduronidase 481:499 Functional recombinant human alpha-L-iduronidase 452:499 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes 452:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 7 14 theme maximum 2038:2044 arg1 yield 2046:2050 a maximum yield 2036:2050 a maximum yield of 1.5% TSP 2036:2062 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 2 15 theme N-glycan 833:840 arg1 maturation 842:851 N-glycan maturation 833:851 N-glycan maturation 833:851 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 0 16 theme mucopolysaccharidosis 160:180 arg1 MESSAGE 189:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 1 17 theme human 475:479 arg1 alpha-L-iduronidase 481:499 Functional recombinant human alpha-L-iduronidase 452:499 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes 452:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 18 theme lysosomal 335:343 arg1 diseases 353:360 lysosomal storage diseases 335:360 lysosomal storage diseases 335:360 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 6 19 theme high-mannose 1893:1904 arg1 N-glycans 1906:1914 the high-mannose N-glycans 1889:1914 the high-mannose N-glycans of gm1-IDUA 1889:1926 This may be due to the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA. 29119347 1 20 from deficient 636:644 arg1 activity 653:660 the activity 649:660 the activity of N-acetylglucosaminyl transferase I 649:698 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 21 theme KEY 185:187 arg1 MESSAGE 189:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 5 22 dep m 1781:1781 arg1 i.e. 1768:1771 i.e. 1768:1771 i.e. 1768:1771 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 1 23 theme N-acetylglucosaminyl 665:684 arg1 transferase 686:696 N-acetylglucosaminyl transferase I 665:698 N-acetylglucosaminyl transferase I 665:698 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 5 24 theme lower 1773:1777 arg1 m 1781:1781 lower K m 1773:1781 lower K m 1773:1781 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 5 24 theme lower 1773:1777 arg1 affinity 1758:1765 a better affinity 1749:1765 a better affinity (i.e. lower K m) for the soluble phosphotransferase 1749:1817 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 0 25 theme human 80:84 arg1 alpha-L-iduronidase 86:104 plant recombinant human alpha-L-iduronidase 62:104 plant recombinant human alpha-L-iduronidase 62:104 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 0 26 theme mannose 382:388 arg1 phosphorylation 390:404 N-glycan mannose phosphorylation 373:404 N-glycan mannose phosphorylation 373:404 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 7 27 theme lines 2023:2027 arg1 screening 2002:2010 screening 2002:2010 screening of the gm1 lines 2002:2027 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 5 28 from cgl-IDUA 1680:1687 arg1 process 1717:1723 the in vitro M6P-tagging process 1692:1723 the in vitro M6P-tagging process 1692:1723 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 4 29 theme human 1526:1530 arg1 UDP-GlcNAc 1552:1561 UDP-GlcNAc 1552:1561 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 4 29 theme human 1526:1530 arg1 phosphotransferase 1532:1549 the human phosphotransferase 1522:1549 the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1522:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 5 30 theme K 1779:1779 arg1 m 1781:1781 lower K m 1773:1781 lower K m 1773:1781 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 5 30 theme K 1779:1779 arg1 affinity 1758:1765 a better affinity 1749:1765 a better affinity (i.e. lower K m) for the soluble phosphotransferase 1749:1817 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 1 31 dep IDUA 502:505 arg1 EC 508:509 EC 3.2.1.76 508:518 IDUA; EC 3.2.1.76 502:518 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 3 32 theme substrate 1172:1180 arg1 4-methylumbelliferyl-iduronide 1182:1211 the artificial substrate 4-methylumbelliferyl-iduronide 1157:1211 the artificial substrate 4-methylumbelliferyl-iduronide 1157:1211 Both forms of IDUA were equivalent with respect to their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide. 29119347 0 33 theme replacement 301:311 arg1 therapeutics 313:324 replacement therapeutics 301:324 replacement therapeutics 301:324 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 5 34 theme soluble 1792:1798 arg1 phosphotransferase 1800:1817 the soluble phosphotransferase 1788:1817 the soluble phosphotransferase 1788:1817 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 0 35 theme N-glycan 0:7 arg1 structures 9:18 N-glycan structures 0:18 N-glycan structures 0:18 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 2 36 gly N-glycosylation 918:932 arg1 enzyme 947:952 the human enzyme 937:952 the human enzyme 937:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 2 37 theme Man5 1003:1006 arg1 N-glycans 976:984 high-mannose N-glycans 963:984 high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures 963:1049 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 4 38 theme enzyme 1574:1579 arg1 -1-phosphotransferase 1609:1629 lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1564:1629 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 7 39 theme soluble 1984:1990 arg1 protein 1992:1998 total soluble protein 1978:1998 total soluble protein 1978:1998 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 7 39 theme soluble 1984:1990 arg1 TSP 1973:1975 > 5.7% TSP 1966:1975 > 5.7% TSP (total soluble protein) 1966:1999 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 4 40 theme [GlcNAc 1601:1607 arg1 -1-phosphotransferase 1609:1629 lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1564:1629 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 6 41 theme residues 1869:1876 arg1 number 1851:1856 the greater number 1839:1856 the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA 1839:1926 This may be due to the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA. 29119347 1 42 theme Arabidopsis 560:570 arg1 background 615:624 the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background 556:624 the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background 556:624 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 43 theme plant 62:66 arg1 alpha-L-iduronidase 86:104 plant recombinant human alpha-L-iduronidase 62:104 plant recombinant human alpha-L-iduronidase 62:104 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 6 44 theme greater 1843:1849 arg1 number 1851:1856 the greater number 1839:1856 the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA 1839:1926 This may be due to the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA. 29119347 7 45 theme elite 1933:1937 arg1 line 1944:1947 Our elite cgl- line 1929:1947 Our elite cgl- line 1929:1947 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 8 46 theme replacement 2191:2201 arg1 therapeutics 2203:2214 enzyme replacement therapeutics 2184:2214 enzyme replacement therapeutics for lysosomal storage diseases 2184:2245 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 2 47 theme N-glycan 871:878 arg1 structures 880:889 the resultant N-glycan structures 857:889 the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme 857:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 1 48 dep alpha-L-iduronidase 481:499 arg1 IDUA 502:505 IDUA 502:505 IDUA; EC 3.2.1.76 502:518 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 5 49 theme M6P-tagging 1705:1715 arg1 process 1717:1723 the in vitro M6P-tagging process 1692:1723 the in vitro M6P-tagging process 1692:1723 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 4 50 theme human 1371:1375 arg1 cells 1377:1381 human cells 1371:1381 human cells 1371:1381 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 6 51 theme mannose 1861:1867 arg1 residues 1869:1876 mannose residues 1861:1876 mannose residues comprising the high-mannose N-glycans of gm1-IDUA 1861:1926 This may be due to the greater number of mannose residues comprising the high-mannose N-glycans of gm1-IDUA. 29119347 1 52 theme I 777:777 arg1 activity 785:792 Golgi α-mannosidase I (GM1) activity 757:792 Golgi α-mannosidase I (GM1) activity 757:792 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 53 theme processing 219:228 arg1 mutants 230:236 Arabidopsis N-glycan processing mutants 198:236 Arabidopsis N-glycan processing mutants 198:236 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 1 54 theme C5 612:613 arg1 background 615:624 the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background 556:624 the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background 556:624 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 7 55 theme > 5.7 1966:1970 arg1 % 1971:1971 % 1971:1971 % 1971:1971 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 8 56 theme storage 2230:2236 arg1 diseases 2238:2245 lysosomal storage diseases 2220:2245 lysosomal storage diseases 2220:2245 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 2 57 theme consensus 898:906 arg1 sites 908:912 the consensus sites 894:912 the consensus sites for N-glycosylation of the human enzyme 894:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 2 58 theme high-mannose 963:974 arg1 N-glycans 976:984 high-mannose N-glycans 963:984 high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures 963:1049 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 3 59 theme IDUA 1066:1069 arg1 forms 1057:1061 Both forms 1052:1061 Both forms of IDUA 1052:1069 Both forms of IDUA were equivalent with respect to their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide. 29119347 2 60 theme enzyme 947:952 arg1 N-glycosylation 918:932 N-glycosylation 918:932 N-glycosylation of the human enzyme 918:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 5 61 theme in 1696:1697 arg1 process 1717:1723 the in vitro M6P-tagging process 1692:1723 the in vitro M6P-tagging process 1692:1723 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 8 62 theme relative 2102:2109 arg1 advantages 2111:2120 the relative advantages 2098:2120 the relative advantages 2098:2120 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 5 63 theme better 1751:1756 arg1 m 1781:1781 lower K m 1773:1781 lower K m 1773:1781 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 5 63 theme better 1751:1756 arg1 affinity 1758:1765 a better affinity 1749:1765 a better affinity (i.e. lower K m) for the soluble phosphotransferase 1749:1817 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 7 64 theme % 1971:1971 arg1 protein 1992:1998 total soluble protein 1978:1998 total soluble protein 1978:1998 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 7 64 theme % 1971:1971 arg1 TSP 1973:1975 > 5.7% TSP 1966:1975 > 5.7% TSP (total soluble protein) 1966:1999 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 7 65 theme TSP 2060:2062 arg1 yield 2046:2050 a maximum yield 2036:2050 a maximum yield of 1.5% TSP 2036:2062 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 1 66 theme Arabidopsis 721:731 arg1 mutant 737:742 the Arabidopsis gm1 mutant 717:742 the Arabidopsis gm1 mutant 717:742 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 7 67 theme 1.5 2055:2057 arg1 % 2058:2058 % 2058:2058 % 2058:2058 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 4 68 theme recombinant 1222:1232 arg1 enzymes 1244:1250 recombinant lysosomal enzymes 1222:1250 recombinant lysosomal enzymes produced in plants 1222:1269 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 1 69 theme mutant 737:742 arg1 seeds 708:712 seeds 708:712 seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity 708:792 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 70 theme I. 182:183 arg1 MESSAGE 189:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 mucopolysaccharidosis I. KEY MESSAGE 160:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 1 71 theme recombinant 463:473 arg1 alpha-L-iduronidase 481:499 Functional recombinant human alpha-L-iduronidase 452:499 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes 452:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 1 72 from activity 653:660 arg1 deficient 636:644 deficient 636:644 deficient 636:644 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 4 73 theme mannose-6-phosphate 1295:1313 arg1 suitable 1336:1343 suitable 1336:1343 suitable 1336:1343 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 4 73 theme mannose-6-phosphate 1295:1313 arg1 addition 1283:1290 the addition 1279:1290 the addition of mannose-6-phosphate (M6P) in order 1279:1328 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 4 74 theme in 1459:1460 arg1 phosphorylation 1476:1490 downstream in vitro mannose phosphorylation 1448:1490 downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1448:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 1 75 theme alpha-L-iduronidase 481:499 arg1 enzymes 521:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes 452:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes 452:527 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 4 76 theme mannose 1468:1474 arg1 phosphorylation 1476:1490 downstream in vitro mannose phosphorylation 1448:1490 downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1448:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 77 theme Arabidopsis 198:208 arg1 mutants 230:236 Arabidopsis N-glycan processing mutants 198:236 Arabidopsis N-glycan processing mutants 198:236 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 0 78 theme storage 345:351 arg1 diseases 353:360 lysosomal storage diseases 335:360 lysosomal storage diseases 335:360 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 1 79 theme transferase 686:696 arg1 activity 653:660 the activity 649:660 the activity of N-acetylglucosaminyl transferase I 649:698 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 4 80 theme phosphotransferase 1532:1549 arg1 form 1514:1517 a soluble form 1504:1517 a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase) 1504:1630 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 81 theme alpha-L-iduronidase 86:104 arg1 structures 9:18 N-glycan structures 0:18 N-glycan structures 0:18 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 0 81 theme alpha-L-iduronidase 86:104 arg1 mannose-phosphorylation 35:57 downstream mannose-phosphorylation 24:57 downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE 24:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 2 82 theme structures 1040:1049 arg1 N-glycans 976:984 high-mannose N-glycans 963:984 high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures 963:1049 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 7 83 theme gm1 2019:2021 arg1 lines 2023:2027 the gm1 lines 2015:2027 the gm1 lines 2015:2027 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 0 84 theme N-glycan 373:380 arg1 phosphorylation 390:404 N-glycan mannose phosphorylation 373:404 N-glycan mannose phosphorylation 373:404 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 2 85 theme gm1-IDUA 1030:1037 arg1 structures 1040:1049 Man6-8 (gm1-IDUA) structures 1022:1049 Man6-8 (gm1-IDUA) structures 1022:1049 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 0 86 theme recombinant 270:280 arg1 enzymes 282:288 recombinant enzymes 270:288 recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy 270:449 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 4 87 theme lysosomal 1349:1357 arg1 delivery 1359:1366 lysosomal delivery 1349:1366 lysosomal delivery in human cells 1349:1381 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 1 88 theme background 615:624 arg1 seeds 547:551 seeds 547:551 seeds 547:551 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 0 89 theme replacement 136:146 arg1 therapy 148:154 enzyme replacement therapy 129:154 enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE 129:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 3 90 theme artificial 1161:1170 arg1 4-methylumbelliferyl-iduronide 1182:1211 the artificial substrate 4-methylumbelliferyl-iduronide 1157:1211 the artificial substrate 4-methylumbelliferyl-iduronide 1157:1211 Both forms of IDUA were equivalent with respect to their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide. 29119347 1 91 dep complex-glycan-deficient 581:604 arg1 cgl 607:609 cgl 607:609 cgl 607:609 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 2 92 from structures 880:889 arg1 sites 908:912 the consensus sites 894:912 the consensus sites for N-glycosylation of the human enzyme 894:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 1 93 theme Golgi 757:761 arg1 I 777:777 Golgi α-mannosidase I 757:777 Golgi α-mannosidase I (GM1) activity 757:792 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 1 93 theme Golgi 757:761 arg1 GM1 780:782 GM1 780:782 GM1 780:782 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 4 94 theme N-acetylglucosamine 1581:1599 arg1 -1-phosphotransferase 1609:1629 lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1564:1629 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 1 95 dep Arabidopsis 560:570 arg1 thaliana 572:579 thaliana 572:579 thaliana 572:579 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 1 95 dep Arabidopsis 560:570 arg1 complex-glycan-deficient 581:604 complex-glycan-deficient 581:604 complex-glycan-deficient 581:604 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 5 96 dep having 1742:1747 arg1 respect 1731:1737 respect 1731:1737 respect 1731:1737 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 0 97 theme downstream 24:33 arg1 mannose-phosphorylation 35:57 downstream mannose-phosphorylation 24:57 downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE 24:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 7 98 theme total 1978:1982 arg1 protein 1992:1998 total soluble protein 1978:1998 total soluble protein 1978:1998 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 7 98 theme total 1978:1982 arg1 TSP 1973:1975 > 5.7% TSP 1966:1975 > 5.7% TSP (total soluble protein) 1966:1999 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 4 99 theme lysosomal 1564:1572 arg1 -1-phosphotransferase 1609:1629 lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1564:1629 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 3 100 theme 4-methylumbelliferyl-iduronide 1182:1211 arg1 cleavage 1145:1152 cleavage 1145:1152 cleavage of the artificial substrate 4-methylumbelliferyl-iduronide 1145:1211 Both forms of IDUA were equivalent with respect to their kinetic parameters characterized by cleavage of the artificial substrate 4-methylumbelliferyl-iduronide. 29119347 2 101 theme Man6-8 1022:1027 arg1 structures 1040:1049 Man6-8 (gm1-IDUA) structures 1022:1049 Man6-8 (gm1-IDUA) structures 1022:1049 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 4 102 from delivery 1359:1366 arg1 cells 1377:1381 human cells 1371:1381 human cells 1371:1381 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 0 103 theme enzyme 129:134 arg1 therapy 148:154 enzyme replacement therapy 129:154 enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE 129:195 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 0 104 theme recombinant 68:78 arg1 alpha-L-iduronidase 86:104 plant recombinant human alpha-L-iduronidase 62:104 plant recombinant human alpha-L-iduronidase 62:104 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 5 105 dep in 1696:1697 arg1 vitro 1699:1703 vitro 1699:1703 vitro 1699:1703 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 8 106 theme enzyme 2184:2189 arg1 therapeutics 2203:2214 enzyme replacement therapeutics 2184:2214 enzyme replacement therapeutics for lysosomal storage diseases 2184:2245 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 4 107 dep UDP-GlcNAc 1552:1561 arg1 -1-phosphotransferase 1609:1629 lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1564:1629 UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase 1552:1629 Because recombinant lysosomal enzymes produced in plants require the addition of mannose-6-phosphate (M6P) in order to be suitable for lysosomal delivery in human cells, we characterized the two IDUA proteins for their amenability to downstream in vitro mannose phosphorylation mediated by a soluble form of the human phosphotransferase (UDP-GlcNAc: lysosomal enzyme N-acetylglucosamine [GlcNAc]-1-phosphotransferase). 29119347 7 108 theme cgl- 1939:1942 arg1 line 1944:1947 Our elite cgl- line 1929:1947 Our elite cgl- line 1929:1947 Our elite cgl- line produces IDUA at > 5.7% TSP (total soluble protein); screening of the gm1 lines showed a maximum yield of 1.5% TSP. 29119347 2 109 theme human 941:945 arg1 enzyme 947:952 the human enzyme 937:952 the human enzyme 937:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 5 110 theme slight 1654:1659 arg1 advantage 1661:1669 a slight advantage 1652:1669 a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process 1652:1723 Gm1-IDUA exhibited a slight advantage over the cgl-IDUA in the in vitro M6P-tagging process, with respect to having a better affinity (i.e. lower K m) for the soluble phosphotransferase. 29119347 0 111 dep structures 9:18 arg1 provide 238:244 provide 238:244 provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy 238:449 N-glycan structures and downstream mannose-phosphorylation of plant recombinant human alpha-L-iduronidase: toward development of enzyme replacement therapy for mucopolysaccharidosis I. KEY MESSAGE: Arabidopsis N-glycan processing mutants provide the basis for tailoring recombinant enzymes for use as replacement therapeutics to treat lysosomal storage diseases, including N-glycan mannose phosphorylation to ensure lysosomal trafficking and efficacy. 29119347 8 112 theme lysosomal 2220:2228 arg1 diseases 2238:2245 lysosomal storage diseases 2220:2245 lysosomal storage diseases 2220:2245 Overall our findings demonstrate the relative advantages and disadvantages associated with the two platforms to create enzyme replacement therapeutics for lysosomal storage diseases. 29119347 2 113 theme resultant 861:869 arg1 structures 880:889 the resultant N-glycan structures 857:889 the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme 857:952 Both strategies effectively prevented N-glycan maturation and the resultant N-glycan structures on the consensus sites for N-glycosylation of the human enzyme revealed high-mannose N-glycans of predominantly Man5 (cgl-IDUA) or Man6-8 (gm1-IDUA) structures. 29119347 1 114 theme α-mannosidase 763:775 arg1 I 777:777 Golgi α-mannosidase I 757:777 Golgi α-mannosidase I (GM1) activity 757:792 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 29119347 1 114 theme α-mannosidase 763:775 arg1 GM1 780:782 GM1 780:782 GM1 780:782 Functional recombinant human alpha-L-iduronidase (IDUA; EC 3.2.1.76) enzymes were generated in seeds of the Arabidopsis thaliana complex-glycan-deficient (cgl) C5 background, which is deficient in the activity of N-acetylglucosaminyl transferase I, and in seeds of the Arabidopsis gm1 mutant, which lacks Golgi α-mannosidase I (GM1) activity. 27617431 7 0 theme 8ANC131-class 1157:1169 arg1 antibodies 1171:1180 VRC01-class and 8ANC131-class antibodies 1141:1180 VRC01-class and 8ANC131-class antibodies 1141:1180 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 7 1 theme VRC01-like 1237:1246 arg1 bNAbs 1248:1252 VRC01-like bNAbs 1237:1252 VRC01-like bNAbs 1237:1252 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 4 2 theme Env 724:726 arg1 trimer 728:733 Env trimer 724:733 Env trimer 724:733 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 6 3 theme CD4bs 1047:1051 arg1 bNAbs 1053:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 1 4 theme structural 145:154 arg1 studies 156:162 structural studies 145:162 structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env) 145:317 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 6 5 contain have 1066:1069 arg2 CDRL3s 1085:1090 normal-length CDRL3s 1071:1090 normal-length CDRL3s 1071:1090 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 6 5 contain have 1066:1069 arg1 bNAbs 1053:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 7 6 contain has 1182:1184 arg2 implications 1186:1197 implications 1186:1197 implications for immunization strategies targeting VRC01-like bNAbs 1186:1252 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 7 6 contain has 1182:1184 arg1 existence 1097:1105 The existence 1093:1105 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies 1093:1180 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 2 7 theme Env 365:367 arg1 glycoforms 369:378 high-mannose and complex-type Env glycoforms 335:378 high-mannose and complex-type Env glycoforms 335:378 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 2 8 from Variability 320:330 arg1 glycoforms 369:378 high-mannose and complex-type Env glycoforms 335:378 high-mannose and complex-type Env glycoforms 335:378 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 0 9 theme CD4-binding 91:101 arg1 site 103:106 the CD4-binding site 87:106 the CD4-binding site 87:106 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 3 10 theme bNAbs 717:721 arg1 epitopes 701:708 complete epitopes 692:708 complete epitopes of two bNAbs 692:721 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 5 11 theme light 932:936 arg1 chain 938:942 its light chain 928:942 its light chain 928:942 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 7 12 theme immunization 1203:1214 arg1 strategies 1216:1225 immunization strategies 1203:1225 immunization strategies targeting VRC01-like bNAbs 1203:1252 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 6 13 link 8ANC131-class/VH1-46-derived 1018:1045 arg1 bNAbs 1053:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 3 14 theme native 576:581 arg1 glycosylation 583:595 fully processed and native glycosylation 556:595 fully processed and native glycosylation 556:595 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 3 15 theme HIV-1 534:538 arg1 trimer 544:549 the HIV-1 Env trimer 530:549 the HIV-1 Env trimer 530:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 2 16 theme complex-type 352:363 arg1 glycoforms 369:378 high-mannose and complex-type Env glycoforms 335:378 high-mannose and complex-type Env glycoforms 335:378 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 3 17 theme 3.9-Å-resolution 491:506 arg1 structures 516:525 3.5-Å- and 3.9-Å-resolution crystal structures 480:525 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer 480:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 5 18 theme CD4bs-targeting 893:907 arg1 bNAbs 921:925 CD4bs-targeting VRC01-class bNAbs 893:925 CD4bs-targeting VRC01-class bNAbs 893:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 4 19 theme site 812:815 arg1 antibody 825:832 a new CD4-binding site (CD4bs) antibody 794:832 a new CD4-binding site (CD4bs) antibody 794:832 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 4 19 theme site 812:815 arg1 IOMA 788:791 IOMA 788:791 IOMA 788:791 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 0 20 theme site 103:106 arg1 recognition 72:82 antibody recognition 63:82 antibody recognition of the CD4-binding site 63:106 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 5 21 theme germline 876:883 arg1 02 868:869 VH1-2*02 862:869 VH1-2*02 862:869 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 21 theme germline 876:883 arg1 gene 885:888 the germline gene 872:888 the germline gene of CD4bs-targeting VRC01-class bNAbs 872:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 2 22 dep glycan 454:459 arg1 shield 461:466 shield 461:466 shield 461:466 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 3 23 theme processed 562:570 arg1 glycosylation 583:595 fully processed and native glycosylation 556:595 fully processed and native glycosylation 556:595 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 0 24 theme glycosylated 9:20 arg1 structure 32:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 5 25 theme bNAbs 921:925 arg1 02 868:869 VH1-2*02 862:869 VH1-2*02 862:869 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 25 theme bNAbs 921:925 arg1 gene 885:888 the germline gene 872:888 the germline gene of CD4bs-targeting VRC01-class bNAbs 872:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 26 theme VH1-2 862:866 arg1 02 868:869 VH1-2*02 862:869 VH1-2*02 862:869 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 26 theme VH1-2 862:866 arg1 gene 885:888 the germline gene 872:888 the germline gene of CD4bs-targeting VRC01-class bNAbs 872:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 1 27 theme trimeric 282:289 arg1 glycoprotein 300:311 the trimeric envelope glycoprotein 278:311 the trimeric envelope glycoprotein (Env) 278:317 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 27 theme trimeric 282:289 arg1 Env 314:316 Env 314:316 Env 314:316 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 0 28 theme new 50:52 arg1 mode 54:57 new mode 50:57 new mode for antibody recognition of the CD4-binding site 50:106 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 5 29 theme * 867:867 arg1 02 868:869 VH1-2*02 862:869 VH1-2*02 862:869 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 29 theme * 867:867 arg1 gene 885:888 the germline gene 872:888 the germline gene of CD4bs-targeting VRC01-class bNAbs 872:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 0 30 theme Env 28:30 arg1 structure 32:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 1 31 theme envelope 291:298 arg1 glycoprotein 300:311 the trimeric envelope glycoprotein 278:311 the trimeric envelope glycoprotein (Env) 278:317 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 31 theme envelope 291:298 arg1 Env 314:316 Env 314:316 Env 314:316 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 4 32 theme CD4bs 818:822 arg1 antibody 825:832 a new CD4-binding site (CD4bs) antibody 794:832 a new CD4-binding site (CD4bs) antibody 794:832 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 4 32 theme CD4bs 818:822 arg1 IOMA 788:791 IOMA 788:791 IOMA 788:791 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 2 33 theme native 447:452 arg1 glycan 454:459 the native glycan shield 443:466 the native glycan shield 443:466 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 2 34 theme glycan 454:459 arg1 visualization 426:438 visualization 426:438 visualization of the native glycan shield 426:466 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 3 35 theme Env 540:542 arg1 trimer 544:549 the HIV-1 Env trimer 530:549 the HIV-1 Env trimer 530:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 1 36 theme neutralizing 204:215 arg1 bNAbs 229:233 bNAbs 229:233 bNAbs 229:233 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 36 theme neutralizing 204:215 arg1 antibodies 217:226 broadly neutralizing antibodies 196:226 broadly neutralizing antibodies (bNAbs) 196:234 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 7 37 theme VRC01-class 1141:1151 arg1 antibodies 1171:1180 VRC01-class and 8ANC131-class antibodies 1141:1180 VRC01-class and 8ANC131-class antibodies 1141:1180 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 3 38 used used 677:680 arg2 we 674:675 we 674:675 we 674:675 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 4 39 theme new 796:798 arg1 antibody 825:832 a new CD4-binding site (CD4bs) antibody 794:832 a new CD4-binding site (CD4bs) antibody 794:832 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 4 39 theme new 796:798 arg1 IOMA 788:791 IOMA 788:791 IOMA 788:791 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 6 40 theme 8ANC131-class/VH1-46-derived 1018:1045 arg1 bNAbs 1053:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 8ANC131-class/VH1-46-derived CD4bs bNAbs 1018:1057 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 6 41 theme normal-length 1071:1083 arg1 CDRL3s 1085:1090 normal-length CDRL3s 1071:1090 normal-length CDRL3s 1071:1090 Thus IOMA resembles 8ANC131-class/VH1-46-derived CD4bs bNAbs, which have normal-length CDRL3s. 27617431 2 42 gly glycoforms 369:378 arg1 Env 365:367 high-mannose and complex-type Env glycoforms 335:378 high-mannose and complex-type Env glycoforms 335:378 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 0 43 gly glycosylated 9:20 arg1 structure 32:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 3 44 theme 3.5-Å- 480:485 arg1 structures 516:525 3.5-Å- and 3.9-Å-resolution crystal structures 480:525 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer 480:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 5 45 theme VRC01-class 979:989 arg1 bNAbs 991:995 VRC01-class bNAbs 979:995 VRC01-class bNAbs 979:995 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 1 46 theme HIV-1 109:113 arg1 design 123:128 HIV-1 vaccine design 109:128 HIV-1 vaccine design 109:128 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 47 from N-glycans 265:273 arg1 glycoprotein 300:311 the trimeric envelope glycoprotein 278:311 the trimeric envelope glycoprotein (Env) 278:317 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 47 from N-glycans 265:273 arg1 Env 314:316 Env 314:316 Env 314:316 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 3 48 theme shield 617:622 arg1 glycan 610:615 a glycan 608:615 a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs 608:721 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 1 49 theme vaccine 115:121 arg1 design 123:128 HIV-1 vaccine design 109:128 HIV-1 vaccine design 109:128 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 3 50 theme complex-type 644:655 arg1 N-glycans 657:665 high-mannose and complex-type N-glycans 627:665 high-mannose and complex-type N-glycans 627:665 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 3 51 theme N-glycans 657:665 arg1 shield 617:622 shield 617:622 shield 617:622 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 3 52 theme high-mannose 627:638 arg1 N-glycans 657:665 high-mannose and complex-type N-glycans 627:665 high-mannose and complex-type N-glycans 627:665 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 1 53 gly glycoprotein 300:311 arg1 glycoprotein 300:311 the trimeric envelope glycoprotein 278:311 the trimeric envelope glycoprotein (Env) 278:317 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 1 53 gly glycoprotein 300:311 arg1 Env 314:316 Env 314:316 Env 314:316 HIV-1 vaccine design is informed by structural studies elucidating mechanisms by which broadly neutralizing antibodies (bNAbs) recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env). 27617431 2 54 theme high-mannose 335:346 arg1 glycoforms 369:378 high-mannose and complex-type Env glycoforms 335:378 high-mannose and complex-type Env glycoforms 335:378 Variability in high-mannose and complex-type Env glycoforms leads to heterogeneity that usually precludes visualization of the native glycan shield. 27617431 5 55 theme short 954:958 arg1 CDRL3 960:964 the short CDRL3 950:964 the short CDRL3 that defines VRC01-class bNAbs 950:995 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 5 56 theme VRC01-class 909:919 arg1 bNAbs 921:925 CD4bs-targeting VRC01-class bNAbs 893:925 CD4bs-targeting VRC01-class bNAbs 893:925 Although IOMA derives from VH1-2*02, the germline gene of CD4bs-targeting VRC01-class bNAbs, its light chain lacks the short CDRL3 that defines VRC01-class bNAbs. 27617431 3 57 theme crystal 508:514 arg1 structures 516:525 3.5-Å- and 3.9-Å-resolution crystal structures 480:525 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer 480:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 0 58 theme antibody 63:70 arg1 recognition 72:82 antibody recognition 63:82 antibody recognition of the CD4-binding site 63:106 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 3 59 theme complete 692:699 arg1 epitopes 701:708 complete epitopes 692:708 complete epitopes of two bNAbs 692:721 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 4 60 theme CD4-binding 800:810 arg1 antibody 825:832 a new CD4-binding site (CD4bs) antibody 794:832 a new CD4-binding site (CD4bs) antibody 794:832 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 4 60 theme CD4-binding 800:810 arg1 IOMA 788:791 IOMA 788:791 IOMA 788:791 Env trimer was complexed with 10-1074 (against the V3-loop) and IOMA, a new CD4-binding site (CD4bs) antibody. 27617431 3 61 theme trimer 544:549 arg1 structures 516:525 3.5-Å- and 3.9-Å-resolution crystal structures 480:525 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer 480:549 We present 3.5-Å- and 3.9-Å-resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation, revealing a glycan shield of high-mannose and complex-type N-glycans, which we used to define complete epitopes of two bNAbs. 27617431 0 62 theme HIV-1 22:26 arg1 structure 32:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure 0:40 Natively glycosylated HIV-1 Env structure reveals new mode for antibody recognition of the CD4-binding site. 27617431 7 63 theme antibodies 1171:1180 arg1 features 1129:1136 features 1129:1136 features of VRC01-class and 8ANC131-class antibodies 1129:1180 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27617431 7 64 theme bNAbs 1110:1114 arg1 existence 1097:1105 The existence 1093:1105 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies 1093:1180 The existence of bNAbs that combine features of VRC01-class and 8ANC131-class antibodies has implications for immunization strategies targeting VRC01-like bNAbs. 27773703 3 0 theme potential 326:334 arg1 sites 352:356 30 potential N-glycosylation sites 323:356 30 potential N-glycosylation sites 323:356 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 6 1 theme ligands 585:591 arg1 absorption 559:568 Kidney absorption 552:568 Kidney absorption of Cy5-labeled ligands 552:591 Kidney absorption of Cy5-labeled ligands was examined in vivo. 27773703 15 2 from megalin 1480:1486 arg1 kidneys 1526:1532 Fut8-/- mouse kidneys 1512:1532 Fut8-/- mouse kidneys 1512:1532 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 13 3 theme higher 1278:1283 arg1 activity 1293:1300 higher binding activity 1278:1300 higher binding activity for RBP and vitamin D-binding protein 1278:1338 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 13 4 contain had 1274:1276 arg2 activity 1293:1300 higher binding activity 1278:1300 higher binding activity for RBP and vitamin D-binding protein 1278:1338 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 13 4 contain had 1274:1276 arg1 megalin 1266:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 3 5 theme N-glycosylation 336:350 arg1 sites 352:356 30 potential N-glycosylation sites 323:356 30 potential N-glycosylation sites 323:356 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 18 6 gly glycoforms 1759:1768 arg1 megalin 1773:1779 megalin 1773:1779 megalin 1773:1779 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 13 7 theme glycoform 1362:1370 arg1 megalin 1372:1378 WGA-positive glycoform megalin 1349:1378 WGA-positive glycoform megalin 1349:1378 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 18 8 theme megalin 1773:1779 arg1 glycoforms 1759:1768 Cell type-specific glycoforms 1740:1768 Cell type-specific glycoforms of megalin 1740:1779 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 8 9 theme convoluted 727:736 arg1 PCTs 747:750 PCTs 747:750 PCTs 747:750 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 9 theme convoluted 727:736 arg1 tubules 738:744 renal proximal convoluted tubules 712:744 renal proximal convoluted tubules (PCTs) 712:751 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 10 10 theme PST 972:974 arg1 megalin 976:982 PST megalin 972:982 PST megalin 972:982 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 10 11 theme wheat 997:1001 arg1 WGA 1020:1022 WGA 1020:1022 WGA 1020:1022 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 10 11 theme wheat 997:1001 arg1 agglutinin 1008:1017 wheat germ agglutinin 997:1017 wheat germ agglutinin (WGA) 997:1023 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 13 12 theme glycoform 1256:1264 arg1 megalin 1266:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 18 13 theme type-specific 1745:1757 arg1 glycoforms 1759:1768 Cell type-specific glycoforms 1740:1768 Cell type-specific glycoforms of megalin 1740:1779 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 15 14 theme RBP-binding 1456:1466 arg1 analysis 1468:1475 RBP-binding analysis 1456:1475 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, 1456:1533 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 6 15 theme Kidney 552:557 arg1 absorption 559:568 Kidney absorption 552:568 Kidney absorption of Cy5-labeled ligands 552:591 Kidney absorption of Cy5-labeled ligands was examined in vivo. 27773703 14 16 theme BSA- 1417:1420 arg1 activities 1444:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 Both glycoforms had nearly the same BSA- and kanamycin-binding activities. 27773703 17 17 from fucosylation 1655:1666 arg1 particular 1672:1681 particular 1672:1681 particular 1672:1681 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 16 18 theme megalin 1600:1606 arg1 N-Glycosylation 1581:1595 N-Glycosylation 1581:1595 N-Glycosylation of megalin 1581:1606 N-Glycosylation of megalin can modulate its ligand-binding activity. 27773703 15 19 theme mouse 1520:1524 arg1 kidneys 1526:1532 Fut8-/- mouse kidneys 1512:1532 Fut8-/- mouse kidneys 1512:1532 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 1 20 gly glycoprotein 115:126 arg1 Megalin 64:70 Megalin 64:70 Megalin 64:70 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 1 20 gly glycoprotein 115:126 arg1 glycoprotein 115:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 7 21 theme ligand 663:668 arg1 blotting 670:677 ligand blotting 663:677 ligand blotting 663:677 Megalin-ligand interactions were analyzed using ligand blotting and ELISA. 27773703 1 22 theme 600-kDa 77:83 arg1 Megalin 64:70 Megalin 64:70 Megalin 64:70 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 1 22 theme 600-kDa 77:83 arg1 glycoprotein 115:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 5 23 theme mouse 537:541 arg1 kidneys 543:549 mouse kidneys 537:549 mouse kidneys 537:549 Distribution of megalin and glycans was histochemically analyzed in mouse kidneys. 27773703 15 24 theme megalin 1480:1486 arg1 analysis 1468:1475 RBP-binding analysis 1456:1475 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, 1456:1533 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 14 25 theme same 1412:1415 arg1 activities 1444:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 Both glycoforms had nearly the same BSA- and kanamycin-binding activities. 27773703 6 26 theme Cy5-labeled 573:583 arg1 ligands 585:591 Cy5-labeled ligands 573:591 Cy5-labeled ligands 573:591 Kidney absorption of Cy5-labeled ligands was examined in vivo. 27773703 15 27 theme Fut8-/- 1512:1518 arg1 kidneys 1526:1532 Fut8-/- mouse kidneys 1512:1532 Fut8-/- mouse kidneys 1512:1532 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 1 28 theme single-spanning 85:99 arg1 Megalin 64:70 Megalin 64:70 Megalin 64:70 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 1 28 theme single-spanning 85:99 arg1 glycoprotein 115:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 3 29 contain has 319:321 arg1 Megalin 311:317 Megalin 311:317 Megalin 311:317 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 3 29 contain has 319:321 arg2 sites 352:356 30 potential N-glycosylation sites 323:356 30 potential N-glycosylation sites 323:356 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 0 30 theme N-Glycoform-dependent 0:20 arg1 interactions 22:33 N-Glycoform-dependent interactions 0:33 N-Glycoform-dependent interactions of megalin with its ligands 0:61 N-Glycoform-dependent interactions of megalin with its ligands. 27773703 8 31 theme straight 766:773 arg1 tubules 775:781 proximal straight tubules 757:781 proximal straight tubules (PSTs) 757:788 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 31 theme straight 766:773 arg1 PSTs 784:787 PSTs 784:787 PSTs 784:787 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 9 32 theme core-fucosyl 891:902 arg1 N-glycans 904:912 core-fucosyl N-glycans 891:912 core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8) 891:956 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 15 33 from analysis 1468:1475 arg1 kidneys 1526:1532 Fut8-/- mouse kidneys 1512:1532 Fut8-/- mouse kidneys 1512:1532 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 1 34 theme transmembrane 101:113 arg1 Megalin 64:70 Megalin 64:70 Megalin 64:70 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 1 34 theme transmembrane 101:113 arg1 glycoprotein 115:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 a 600-kDa single-spanning transmembrane glycoprotein 75:126 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 17 35 theme megalin-RBP 1714:1724 arg1 interactions 1726:1737 megalin-RBP interactions 1714:1737 megalin-RBP interactions 1714:1737 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 0 36 theme megalin 38:44 arg1 interactions 22:33 N-Glycoform-dependent interactions 0:33 N-Glycoform-dependent interactions of megalin with its ligands 0:61 N-Glycoform-dependent interactions of megalin with its ligands. 27773703 8 37 theme proximal 757:764 arg1 tubules 775:781 proximal straight tubules 757:781 proximal straight tubules (PSTs) 757:788 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 37 theme proximal 757:764 arg1 PSTs 784:787 PSTs 784:787 PSTs 784:787 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 15 38 from kidneys 1526:1532 arg1 analysis 1468:1475 RBP-binding analysis 1456:1475 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, 1456:1533 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 15 39 contain had 1535:1537 arg2 activity 1571:1578 significantly decreased binding activity 1539:1578 significantly decreased binding activity 1539:1578 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 15 39 contain had 1535:1537 arg1 analysis 1468:1475 RBP-binding analysis 1456:1475 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, 1456:1533 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 8 40 theme renal 712:716 arg1 PCTs 747:750 PCTs 747:750 PCTs 747:750 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 40 theme renal 712:716 arg1 tubules 738:744 renal proximal convoluted tubules 712:744 renal proximal convoluted tubules (PCTs) 712:751 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 5 41 theme glycans 497:503 arg1 Distribution 469:480 Distribution 469:480 Distribution of megalin and glycans 469:503 Distribution of megalin and glycans was histochemically analyzed in mouse kidneys. 27773703 7 42 theme Megalin-ligand 615:628 arg1 interactions 630:641 Megalin-ligand interactions 615:641 Megalin-ligand interactions 615:641 Megalin-ligand interactions were analyzed using ligand blotting and ELISA. 27773703 13 43 theme purified 1234:1241 arg1 megalin 1266:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 15 44 theme binding 1563:1569 arg1 activity 1571:1578 significantly decreased binding activity 1539:1578 significantly decreased binding activity 1539:1578 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 18 45 theme tubular 1803:1809 arg1 cells 1811:1815 the proximal tubular cells 1790:1815 the proximal tubular cells 1790:1815 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 15 46 theme core 1496:1499 arg1 fucose 1501:1506 core fucose 1496:1506 core fucose 1496:1506 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 17 47 theme Core 1650:1653 arg1 fucosylation 1655:1666 Core fucosylation 1650:1666 Core fucosylation 1650:1666 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 17 47 theme Core 1650:1653 arg1 modification 1689:1700 a modification 1687:1700 a modification crucial for megalin-RBP interactions 1687:1737 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 13 48 theme WGA-positive 1349:1360 arg1 megalin 1372:1378 WGA-positive glycoform megalin 1349:1378 WGA-positive glycoform megalin 1349:1378 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 8 49 contain have 790:793 arg1 Megalins 690:697 Megalins 690:697 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) 690:788 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 49 contain have 790:793 arg2 N-glycans 805:813 different N-glycans 795:813 different N-glycans 795:813 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 1 50 theme other 215:219 arg1 tissues 221:227 other tissues 215:227 other tissues 215:227 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 18 51 theme absorption 1837:1846 arg1 capacity 1848:1855 ligand absorption capacity 1830:1855 ligand absorption capacity 1830:1855 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 10 52 theme hybrid-type 1043:1053 arg1 N-glycans 1055:1063 hybrid-type N-glycans 1043:1063 hybrid-type N-glycans 1043:1063 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 9 53 theme PCT 816:818 arg1 megalin 820:826 PCT megalin 816:826 PCT megalin 816:826 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 18 54 theme ligand 1830:1835 arg1 capacity 1848:1855 ligand absorption capacity 1830:1855 ligand absorption capacity 1830:1855 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 8 55 theme different 795:803 arg1 N-glycans 805:813 different N-glycans 795:813 different N-glycans 795:813 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 3 56 theme extracellular 365:377 arg1 domain 379:384 its extracellular domain 361:384 its extracellular domain 361:384 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 10 57 theme germ 1003:1006 arg1 WGA 1020:1022 WGA 1020:1022 WGA 1020:1022 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 10 57 theme germ 1003:1006 arg1 agglutinin 1008:1017 wheat germ agglutinin 997:1017 wheat germ agglutinin (WGA) 997:1023 In contrast, PST megalin stained with wheat germ agglutinin (WGA), which recognizes hybrid-type N-glycans. 27773703 0 58 with interactions 22:33 arg1 ligands 55:61 its ligands 51:61 its ligands 51:61 N-Glycoform-dependent interactions of megalin with its ligands. 27773703 11 59 theme Retinol-binding 1066:1080 arg1 RBP-Cy5 1095:1101 RBP-Cy5 1095:1101 RBP-Cy5 1095:1101 Retinol-binding protein-Cy5 (RBP-Cy5) was endocytosed by megalin on PCTs but minimally endocytosed by PSTs. 27773703 11 59 theme Retinol-binding 1066:1080 arg1 protein-Cy5 1082:1092 Retinol-binding protein-Cy5 1066:1092 Retinol-binding protein-Cy5 (RBP-Cy5) 1066:1102 Retinol-binding protein-Cy5 (RBP-Cy5) was endocytosed by megalin on PCTs but minimally endocytosed by PSTs. 27773703 14 60 contain had 1397:1399 arg1 glycoforms 1386:1395 Both glycoforms 1381:1395 Both glycoforms 1381:1395 Both glycoforms had nearly the same BSA- and kanamycin-binding activities. 27773703 14 60 contain had 1397:1399 arg2 activities 1444:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 Both glycoforms had nearly the same BSA- and kanamycin-binding activities. 27773703 16 61 theme ligand-binding 1625:1638 arg1 activity 1640:1647 its ligand-binding activity 1621:1647 its ligand-binding activity 1621:1647 N-Glycosylation of megalin can modulate its ligand-binding activity. 27773703 3 62 gly N-glycosylation 336:350 arg2 30 323:324 30 323:324 30 323:324 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 3 62 gly N-glycosylation 336:350 arg2 sites 352:356 30 potential N-glycosylation sites 323:356 30 potential N-glycosylation sites 323:356 Megalin has 30 potential N-glycosylation sites in its extracellular domain. 27773703 9 63 theme Lens 841:844 arg1 LCA 868:870 LCA 868:870 LCA 868:870 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 9 63 theme Lens 841:844 arg1 agglutinin 856:865 Lens culinaris agglutinin 841:865 Lens culinaris agglutinin (LCA) 841:871 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 13 64 theme LCA-positive 1243:1254 arg1 megalin 1266:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin 1230:1272 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 14 65 theme kanamycin-binding 1426:1442 arg1 activities 1444:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 nearly the same BSA- and kanamycin-binding activities 1401:1453 Both glycoforms had nearly the same BSA- and kanamycin-binding activities. 27773703 16 66 gly N-Glycosylation 1581:1595 arg1 megalin 1600:1606 megalin 1600:1606 megalin 1600:1606 N-Glycosylation of megalin can modulate its ligand-binding activity. 27773703 17 67 gly fucosylation 1655:1666 arg1 particular 1672:1681 particular 1672:1681 particular 1672:1681 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 13 68 theme D-binding 1322:1330 arg1 protein 1332:1338 vitamin D-binding protein 1314:1338 vitamin D-binding protein 1314:1338 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 1 69 theme endocytic 148:156 arg1 Megalin 64:70 Megalin 64:70 Megalin 64:70 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 1 69 theme endocytic 148:156 arg1 receptor 158:165 an endocytic receptor 145:165 an endocytic receptor 145:165 Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. 27773703 17 70 theme crucial 1702:1708 arg1 fucosylation 1655:1666 Core fucosylation 1650:1666 Core fucosylation 1650:1666 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 17 70 theme crucial 1702:1708 arg1 modification 1689:1700 a modification 1687:1700 a modification crucial for megalin-RBP interactions 1687:1737 Core fucosylation, in particular, is a modification crucial for megalin-RBP interactions. 27773703 4 71 theme glycoform-dependent 441:459 arg1 manner 461:466 a glycoform-dependent manner 439:466 a glycoform-dependent manner 439:466 We found that megalin interacts with its ligands in a glycoform-dependent manner. 27773703 2 72 theme distinct 260:267 arg1 ligands 269:275 Structurally and functionally distinct ligands 230:275 Structurally and functionally distinct ligands for megalin 230:287 Structurally and functionally distinct ligands for megalin have been identified. 27773703 13 73 theme binding 1285:1291 arg1 activity 1293:1300 higher binding activity 1278:1300 higher binding activity for RBP and vitamin D-binding protein 1278:1338 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 27773703 5 74 theme megalin 485:491 arg1 Distribution 469:480 Distribution 469:480 Distribution of megalin and glycans 469:503 Distribution of megalin and glycans was histochemically analyzed in mouse kidneys. 27773703 15 75 theme decreased 1553:1561 arg1 activity 1571:1578 significantly decreased binding activity 1539:1578 significantly decreased binding activity 1539:1578 RBP-binding analysis of megalin lacking core fucose, in Fut8-/- mouse kidneys, had significantly decreased binding activity. 27773703 8 76 theme proximal 718:725 arg1 PCTs 747:750 PCTs 747:750 PCTs 747:750 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 8 76 theme proximal 718:725 arg1 tubules 738:744 renal proximal convoluted tubules 712:744 renal proximal convoluted tubules (PCTs) 712:751 Megalins expressed on renal proximal convoluted tubules (PCTs) and proximal straight tubules (PSTs) have different N-glycans. 27773703 9 77 theme culinaris 846:854 arg1 LCA 868:870 LCA 868:870 LCA 868:870 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 9 77 theme culinaris 846:854 arg1 agglutinin 856:865 Lens culinaris agglutinin 841:865 Lens culinaris agglutinin (LCA) 841:871 PCT megalin stained with Lens culinaris agglutinin (LCA), which recognizes core-fucosyl N-glycans catalyzed by α1,6-fucosyltransferase (Fut8). 27773703 18 78 theme proximal 1794:1801 arg1 cells 1811:1815 the proximal tubular cells 1790:1815 the proximal tubular cells 1790:1815 Cell type-specific glycoforms of megalin exist in the proximal tubular cells and modulate ligand absorption capacity. 27773703 13 79 dep did 1345:1347 arg1 than 1340:1343 than 1340:1343 than 1340:1343 The purified LCA-positive glycoform megalin had higher binding activity for RBP and vitamin D-binding protein than did WGA-positive glycoform megalin. 26088564 9 0 theme AGP-ligand-binding 1266:1283 arg1 site 1285:1288 the AGP-ligand-binding site 1262:1288 the AGP-ligand-binding site 1262:1288 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 1 1 gly glycoprotein 175:186 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 1 gly glycoprotein 175:186 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 1 gly glycoprotein 175:186 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 7 2 theme glycosylated 931:942 arg1 form 944:947 its fully glycosylated form 921:947 its fully glycosylated form 921:947 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 1 3 theme abundant 153:160 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 3 theme abundant 153:160 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 3 theme abundant 153:160 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 6 4 theme AGP 606:608 arg1 structure 578:586 the X-ray structure 568:586 the X-ray structure of unglycosylated AGP 568:608 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 8 5 theme influence 1070:1078 arg1 absence 1051:1057 the absence 1047:1057 the absence of a major influence of glycosylation on AGP's secondary structure 1047:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 8 6 from structure 1116:1124 arg1 absence 1051:1057 the absence 1047:1057 the absence of a major influence of glycosylation on AGP's secondary structure 1047:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 7 7 dep glycobiology 854:865 arg1 the 839:841 the 839:841 the 839:841 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 1 8 theme human 162:166 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 8 theme human 162:166 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 8 theme human 162:166 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 6 9 theme unglycosylated 591:604 arg1 AGP 606:608 unglycosylated AGP 591:608 unglycosylated AGP 591:608 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 7 10 theme form 944:947 arg1 model 912:916 a structural model 899:916 a structural model of its fully glycosylated form 899:947 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 6 11 gly unglycosylated 591:604 arg1 AGP 606:608 unglycosylated AGP 591:608 unglycosylated AGP 591:608 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 11 12 theme increased 1658:1666 arg1 participation 1668:1680 an increased participation 1655:1680 an increased participation of carbohydrates 1655:1697 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 8 13 with agreement 1130:1138 arg1 observations 1161:1172 crystallography observations 1145:1172 crystallography observations 1145:1172 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 3 14 theme glycan 350:355 arg1 chains 357:362 the glycan chains 346:362 the glycan chains attached to Asn54, Asn75 and Asn85 346:397 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 9 15 theme protein 1234:1240 arg1 dynamics 1242:1249 the protein dynamics 1230:1249 the protein dynamics 1230:1249 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 7 16 theme structural 901:910 arg1 model 912:916 a structural model 899:916 a structural model of its fully glycosylated form 899:947 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 10 17 theme AGP 1424:1426 arg1 structure 1428:1436 AGP structure 1424:1436 AGP structure 1424:1436 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 6 18 theme further 675:681 arg1 insights 683:690 the further insights 671:690 the further insights into its structural biology and, ultimately, into its biological function 671:764 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 9 19 theme other 1351:1355 arg1 compounds 1367:1375 other bioactive compounds 1351:1375 other bioactive compounds 1351:1375 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 1 20 gly N-glycosylated 200:213 arg2 positions 223:231 five positions 218:231 five positions 218:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 20 gly N-glycosylated 200:213 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 20 gly N-glycosylated 200:213 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 20 gly N-glycosylated 200:213 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 7 21 theme structural 843:852 arg1 glycobiology 854:865 structural glycobiology 843:865 structural glycobiology 843:865 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 1 22 theme plasma 168:173 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 22 theme plasma 168:173 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 22 theme plasma 168:173 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 6 23 theme biological 746:755 arg1 function 757:764 its biological function 742:764 its biological function 742:764 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 6 24 theme X-ray 572:576 arg1 structure 578:586 the X-ray structure 568:586 the X-ray structure of unglycosylated AGP 568:608 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 7 25 theme different 974:982 arg1 glycoforms 984:993 the different glycoforms 970:993 the different glycoforms that are found in vivo 970:1016 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 8 26 theme secondary 1106:1114 arg1 structure 1116:1124 AGP's secondary structure 1100:1124 AGP's secondary structure 1100:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 10 27 theme glycan 1507:1512 arg1 chains 1514:1519 glycan chains 1507:1519 glycan chains linked to Asn54 and Asn75 1507:1545 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 9 28 from role 1313:1316 arg1 complexation 1325:1336 its complexation 1321:1336 its complexation to drugs and other bioactive compounds 1321:1375 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 2 29 theme inflammatory 309:320 arg1 events 322:327 inflammatory events 309:327 inflammatory events 309:327 AGP plays important roles on pharmacokinetics and can rise up to 5-fold in inflammatory events. 26088564 6 30 theme chains 655:660 arg1 absence 633:639 the absence 629:639 the absence of the glycan chains 629:660 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 0 31 theme Structural 0:9 arg1 glycobiology 11:22 Structural glycobiology 0:22 Structural glycobiology of human α1-acid glycoprotein 0:52 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 7 32 theme AGP 883:885 arg1 function 871:878 function 871:878 function 871:878 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 7 32 theme AGP 883:885 arg1 glycobiology 854:865 structural glycobiology 843:865 structural glycobiology 843:865 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 4 33 from epitope 466:472 arg1 turn 478:481 turn 478:481 turn 478:481 This epitope, in turn, can bind selectin proteins. 26088564 6 34 theme glycan 648:653 arg1 chains 655:660 the glycan chains 644:660 the glycan chains 644:660 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 0 35 theme human 27:31 arg1 glycoprotein 41:52 human α1-acid glycoprotein 27:52 human α1-acid glycoprotein 27:52 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 10 36 theme fucosylation 1408:1419 arg1 influence 1395:1403 the influence 1391:1403 the influence of fucosylation on AGP structure and binding to selectins 1391:1461 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 8 37 theme obtained 1023:1030 arg1 data 1032:1035 The obtained data 1019:1035 The obtained data 1019:1035 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 9 38 theme bioactive 1357:1365 arg1 compounds 1367:1375 other bioactive compounds 1351:1375 other bioactive compounds 1351:1375 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 1 39 theme Human 114:118 arg1 AGP 142:144 AGP 142:144 AGP 142:144 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 39 theme Human 114:118 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 39 theme Human 114:118 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 39 theme Human 114:118 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 3 40 theme sialyl-Lewis 437:448 arg1 epitope 452:458 a sialyl-Lewis X epitope 435:458 a sialyl-Lewis X epitope 435:458 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 7 41 gly glycosylated 931:942 arg1 form 944:947 its fully glycosylated form 921:947 its fully glycosylated form 921:947 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 1 42 gly glycoprotein 128:139 arg1 AGP 142:144 AGP 142:144 AGP 142:144 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 42 gly glycoprotein 128:139 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 42 gly glycoprotein 128:139 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 42 gly glycoprotein 128:139 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 43 theme α1-acid 120:126 arg1 AGP 142:144 AGP 142:144 AGP 142:144 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 43 theme α1-acid 120:126 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 43 theme α1-acid 120:126 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 43 theme α1-acid 120:126 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 10 44 gly fucosylation 1408:1419 arg1 AGP 1424:1426 AGP structure 1424:1436 AGP structure 1424:1436 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 3 45 theme X 450:450 arg1 epitope 452:458 a sialyl-Lewis X epitope 435:458 a sialyl-Lewis X epitope 435:458 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 0 46 theme glycoprotein 41:52 arg1 implications 62:73 its implications 58:73 its implications for pharmacokinetics and inflammation 58:111 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 0 46 theme glycoprotein 41:52 arg1 glycobiology 11:22 Structural glycobiology 0:22 Structural glycobiology of human α1-acid glycoprotein 0:52 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 8 47 from influence 1070:1078 arg1 structure 1116:1124 AGP's secondary structure 1100:1124 AGP's secondary structure 1100:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 7 48 theme current 777:783 arg1 work 785:788 the current work 773:788 the current work 773:788 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 0 49 theme α1-acid 33:39 arg1 glycoprotein 41:52 human α1-acid glycoprotein 27:52 human α1-acid glycoprotein 27:52 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 11 50 theme observed 1706:1713 arg1 roles 1719:1723 the observed AGP roles 1702:1723 the observed AGP roles in pharmacokinetics and inflammation 1702:1760 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 8 51 from absence 1051:1057 arg1 structure 1116:1124 AGP's secondary structure 1100:1124 AGP's secondary structure 1100:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 10 52 attach attached 1613:1620 arg2 one 1609:1611 the one 1605:1611 the one attached to Asn15 1605:1629 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 10 52 attach attached 1613:1620 arg1 Asn15 1625:1629 Asn15 1625:1629 Asn15 1625:1629 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 9 53 theme possible 1304:1311 arg1 role 1313:1316 a possible role 1302:1316 a possible role in its complexation to drugs and other bioactive compounds 1302:1375 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 10 54 from influence 1395:1403 arg1 binding 1442:1448 binding 1442:1448 binding to selectins 1442:1461 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 10 54 from influence 1395:1403 arg1 structure 1428:1436 AGP structure 1424:1436 AGP structure 1424:1436 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 11 55 theme AGP 1715:1717 arg1 roles 1719:1723 the observed AGP roles 1702:1723 the observed AGP roles in pharmacokinetics and inflammation 1702:1760 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 11 56 theme carbohydrates 1685:1697 arg1 participation 1668:1680 an increased participation 1655:1680 an increased participation of carbohydrates 1655:1697 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 10 57 attach linked 1521:1526 arg1 Asn54 1531:1535 Asn54 1531:1535 Asn54 1531:1535 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 10 57 attach linked 1521:1526 arg1 Asn75 1541:1545 Asn75 1541:1545 Asn75 1541:1545 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 10 57 attach linked 1521:1526 arg2 chains 1514:1519 glycan chains 1507:1519 glycan chains linked to Asn54 and Asn75 1507:1545 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 5 58 theme Such 512:515 arg1 interplay 517:525 Such interplay 512:525 Such interplay 512:525 Such interplay is important for immunomodulation. 26088564 8 59 theme crystallography 1145:1159 arg1 observations 1161:1172 crystallography observations 1145:1172 crystallography observations 1145:1172 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 9 60 theme glycan 1188:1193 arg1 chains 1195:1200 the glycan chains 1184:1200 the glycan chains 1184:1200 However, the glycan chains seem able to interfere with the protein dynamics, mainly at the AGP-ligand-binding site, indicating a possible role in its complexation to drugs and other bioactive compounds. 26088564 7 61 theme function 871:878 arg1 characterization 819:834 the characterization 815:834 the characterization of the structural glycobiology and function of AGP 815:885 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 4 62 theme selectin 493:500 arg1 proteins 502:509 selectin proteins 493:509 selectin proteins 493:509 This epitope, in turn, can bind selectin proteins. 26088564 1 63 from positions 223:231 arg1 N-glycosylated 200:213 N-glycosylated 200:213 N-glycosylated 200:213 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 63 from positions 223:231 arg1 glycoprotein 128:139 Human α1-acid glycoprotein 114:139 Human α1-acid glycoprotein (AGP) 114:145 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 1 63 from positions 223:231 arg1 glycoprotein 175:186 an abundant human plasma glycoprotein 150:186 an abundant human plasma glycoprotein that may be N-glycosylated at five positions 150:231 Human α1-acid glycoprotein (AGP) is an abundant human plasma glycoprotein that may be N-glycosylated at five positions. 26088564 11 64 from roles 1719:1723 arg1 pharmacokinetics 1728:1743 pharmacokinetics 1728:1743 pharmacokinetics 1728:1743 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 11 64 from roles 1719:1723 arg1 inflammation 1749:1760 inflammation 1749:1760 inflammation 1749:1760 These results point to an increased participation of carbohydrates on the observed AGP roles in pharmacokinetics and inflammation. 26088564 3 65 attach attached 364:371 arg2 chains 357:362 the glycan chains 346:362 the glycan chains attached to Asn54, Asn75 and Asn85 346:397 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 3 65 attach attached 364:371 arg1 Asn54 376:380 Asn54 376:380 Asn54 376:380 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 3 65 attach attached 364:371 arg1 Asn85 393:397 Asn85 393:397 Asn85 393:397 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 3 65 attach attached 364:371 arg1 Asn75 383:387 Asn75 383:387 Asn75 383:387 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 0 66 gly glycoprotein 41:52 arg1 glycoprotein 41:52 human α1-acid glycoprotein 27:52 human α1-acid glycoprotein 27:52 Structural glycobiology of human α1-acid glycoprotein and its implications for pharmacokinetics and inflammation. 26088564 6 67 theme structural 701:710 arg1 biology 712:718 its structural biology 697:718 its structural biology 697:718 While the X-ray structure of unglycosylated AGP has been reported, the absence of the glycan chains hampered the further insights into its structural biology and, ultimately, into its biological function. 26088564 10 68 theme other 1582:1586 arg1 glycans 1588:1594 other glycans 1582:1594 other glycans 1582:1594 By examining the influence of fucosylation on AGP structure and binding to selectins, it is proposed that the latter may bind to glycan chains linked to Asn54 and Asn75, and that this binding may involve other glycans, such as the one attached to Asn15. 26088564 7 69 theme glycobiology 854:865 arg1 characterization 819:834 the characterization 815:834 the characterization of the structural glycobiology and function of AGP 815:885 Thus, the current work intends to contribute in the characterization of the structural glycobiology and function of AGP by building a structural model of its fully glycosylated form, taking into account the different glycoforms that are found in vivo. 26088564 8 70 theme major 1064:1068 arg1 influence 1070:1078 a major influence 1062:1078 a major influence of glycosylation on AGP's secondary structure 1062:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 26088564 3 71 theme such 333:336 arg1 events 338:343 such events 333:343 such events 333:343 In such events, the glycan chains attached to Asn54, Asn75 and Asn85 may become fucosylated, originating a sialyl-Lewis X epitope. 26088564 2 72 theme important 244:252 arg1 roles 254:258 important roles 244:258 important roles 244:258 AGP plays important roles on pharmacokinetics and can rise up to 5-fold in inflammatory events. 26088564 8 73 theme glycosylation 1083:1095 arg1 influence 1070:1078 a major influence 1062:1078 a major influence of glycosylation on AGP's secondary structure 1062:1124 The obtained data points to the absence of a major influence of glycosylation on AGP's secondary structure, in agreement with crystallography observations. 25700513 3 0 theme antiparallel 302:313 arg1 orientation 323:333 a two-site, antiparallel binding orientation 290:333 orientation 323:333 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 3 1 theme crystal 210:216 arg1 structure 218:226 The 2.3 angstrom resolution crystal structure 182:226 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex 182:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 5 2 from residues 585:592 arg1 Notch1 597:602 Notch1 597:602 Notch1 597:602 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 3 3 theme binding 315:321 arg1 orientation 323:333 a two-site, antiparallel binding orientation 290:333 orientation 323:333 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 2 4 theme fate 113:116 arg1 decisions 118:126 mammalian cell fate decisions 98:126 mammalian cell fate decisions 98:126 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 4 5 from N-terminus 506:515 arg1 MNNL 535:538 MNNL 535:538 MNNL 535:538 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 4 5 from N-terminus 506:515 arg1 module 492:497 module 492:497 module at the N-terminus of Notch ligands 492:532 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 4 5 from N-terminus 506:515 arg1 domain 481:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 6 6 theme role 789:792 arg1 elucidation 756:766 The elucidation 752:766 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement 752:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 3 7 theme O-linked 354:361 arg1 glycosylation 363:375 Notch1 O-linked glycosylation 347:375 Notch1 O-linked glycosylation 347:375 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 2 8 theme cell 108:111 arg1 decisions 118:126 mammalian cell fate decisions 98:126 mammalian cell fate decisions 98:126 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 3 9 theme angstrom 190:197 arg1 resolution 199:208 2.3 angstrom resolution 186:208 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex 182:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 4 10 theme factor-like 402:412 arg1 repeats 414:420 Notch1 epidermal growth factor-like repeats 378:420 Notch1 epidermal growth factor-like repeats 11 and 12 378:430 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 6 11 theme functional 964:973 arg1 capacity 975:982 their functional capacity 958:982 their functional capacity 958:982 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 5 12 theme essential 711:719 arg1 contacts 722:729 specific, and essential, contacts 697:729 specific, and essential, contacts 697:729 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 4 13 theme domain 481:486 arg1 domains 541:547 the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains 446:547 the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains 446:547 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 3 14 theme resolution 199:208 arg1 structure 218:226 The 2.3 angstrom resolution crystal structure 182:226 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex 182:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 6 15 theme direct 773:778 arg1 role 789:792 a direct chemical role 771:792 a direct chemical role for O-glycans in Notch1 ligand engagement 771:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 5 16 theme serine 578:583 arg1 residues 585:592 Threonine and serine residues 564:592 Threonine and serine residues on Notch1 564:602 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 3 17 theme interacting 235:245 arg1 regions 247:253 the interacting regions 231:253 the interacting regions of the Notch1-DLL4 complex 231:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 1 18 theme 4 73:73 arg1 engagement 48:57 Notch1 engagement 41:57 Notch1 engagement of Delta-like 4 41:73 Structural basis for Notch1 engagement of Delta-like 4. 25700513 3 19 theme Notch1 347:352 arg1 glycosylation 363:375 Notch1 O-linked glycosylation 347:375 Notch1 O-linked glycosylation 347:375 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 4 20 theme module 492:497 arg1 domains 541:547 the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains 446:547 the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains 446:547 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 3 21 theme regions 247:253 arg1 structure 218:226 The 2.3 angstrom resolution crystal structure 182:226 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex 182:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 4 22 theme Notch1 378:383 arg1 repeats 414:420 Notch1 epidermal growth factor-like repeats 378:420 Notch1 epidermal growth factor-like repeats 11 and 12 378:430 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 6 23 from engagement 825:834 arg1 elucidation 756:766 The elucidation 752:766 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement 752:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 6 24 theme Notch 931:935 arg1 proteins 937:944 Notch proteins 931:944 Notch proteins 931:944 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 5 25 theme Threonine 564:572 arg1 residues 585:592 Threonine and serine residues 564:592 Threonine and serine residues on Notch1 564:602 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 6 26 from role 789:792 arg1 engagement 825:834 Notch1 ligand engagement 811:834 Notch1 ligand engagement 811:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 0 27 theme Structural 0:9 arg1 biology 11:17 Structural biology 0:17 Structural biology. 0:18 Structural biology. 25700513 4 28 theme growth 395:400 arg1 repeats 414:420 Notch1 epidermal growth factor-like repeats 378:420 Notch1 epidermal growth factor-like repeats 11 and 12 378:430 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 3 29 theme Notch1-DLL4 262:272 arg1 complex 274:280 the Notch1-DLL4 complex 258:280 the Notch1-DLL4 complex 258:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 4 30 theme epidermal 385:393 arg1 repeats 414:420 Notch1 epidermal growth factor-like repeats 378:420 Notch1 epidermal growth factor-like repeats 11 and 12 378:430 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 3 31 link O-linked 354:361 arg1 glycosylation 363:375 Notch1 O-linked glycosylation 347:375 Notch1 O-linked glycosylation 347:375 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 4 32 theme Delta/Serrate/Lag-2 455:473 arg1 MNNL 535:538 MNNL 535:538 MNNL 535:538 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 4 32 theme Delta/Serrate/Lag-2 455:473 arg1 domain 481:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 2 33 dep proteins 144:151 arg1 DLL 176:178 DLL 176:178 DLL 176:178 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 2 33 dep proteins 144:151 arg1 Delta-like 164:173 Delta-like 164:173 Delta-like 164:173 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 2 33 dep proteins 144:151 arg1 Jagged 153:158 Jagged 153:158 Jagged 153:158 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 5 34 theme specific 697:704 arg1 contacts 722:729 specific, and essential, contacts 697:729 specific, and essential, contacts 697:729 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 4 35 theme DLL4 450:453 arg1 MNNL 535:538 MNNL 535:538 MNNL 535:538 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 4 35 theme DLL4 450:453 arg1 domain 481:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 2 36 theme mammalian 98:106 arg1 decisions 118:126 mammalian cell fate decisions 98:126 mammalian cell fate decisions 98:126 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 4 37 theme DSL 476:478 arg1 MNNL 535:538 MNNL 535:538 MNNL 535:538 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 4 37 theme DSL 476:478 arg1 domain 481:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 DLL4 Delta/Serrate/Lag-2 (DSL) domain 450:486 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 6 38 theme ligand 909:914 arg1 sites 924:928 their ligand binding sites 903:928 their ligand binding sites 903:928 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 4 39 theme Notch 520:524 arg1 ligands 526:532 Notch ligands 520:532 Notch ligands 520:532 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 3 40 theme two-site 292:299 arg1 orientation 323:333 a two-site, antiparallel binding orientation 290:333 orientation 323:333 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 6 41 theme ligand 818:823 arg1 engagement 825:834 Notch1 ligand engagement 811:834 Notch1 ligand engagement 811:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 4 42 theme ligands 526:532 arg1 N-terminus 506:515 the N-terminus 502:515 the N-terminus of Notch ligands 502:532 Notch1 epidermal growth factor-like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. 25700513 6 43 theme posttranslational 868:884 arg1 modifications 886:898 posttranslational modifications 868:898 posttranslational modifications of their ligand binding sites 868:928 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 6 44 theme binding 916:922 arg1 sites 924:928 their ligand binding sites 903:928 their ligand binding sites 903:928 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 5 45 theme surrogate 665:673 arg1 O-fucose 628:635 O-fucose 628:635 O-fucose 628:635 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 5 45 theme surrogate 665:673 arg1 acids 681:685 surrogate amino acids 665:685 surrogate amino acids 665:685 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 5 45 theme surrogate 665:673 arg1 O-glucose 641:649 O-glucose 641:649 O-glucose 641:649 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 6 46 theme Notch1 811:816 arg1 engagement 825:834 Notch1 ligand engagement 811:834 Notch1 ligand engagement 811:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 1 47 theme Structural 20:29 arg1 basis 31:35 Structural basis 20:35 Structural basis for Notch1 engagement of Delta-like 4. 20:74 Structural basis for Notch1 engagement of Delta-like 4. 25700513 6 48 from elucidation 756:766 arg1 engagement 825:834 Notch1 ligand engagement 811:834 Notch1 ligand engagement 811:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 2 49 theme Notch 76:80 arg1 receptors 82:90 Notch receptors 76:90 Notch receptors 76:90 Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). 25700513 5 50 theme amino 675:679 arg1 O-fucose 628:635 O-fucose 628:635 O-fucose 628:635 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 5 50 theme amino 675:679 arg1 acids 681:685 surrogate amino acids 665:685 surrogate amino acids 665:685 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 5 50 theme amino 675:679 arg1 O-glucose 641:649 O-glucose 641:649 O-glucose 641:649 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 6 51 theme chemical 780:787 arg1 role 789:792 a direct chemical role 771:792 a direct chemical role for O-glycans in Notch1 ligand engagement 771:834 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 6 52 attach linked 951:956 arg2 proteins 937:944 Notch proteins 931:944 Notch proteins 931:944 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 6 52 attach linked 951:956 arg1 pathways 1026:1033 developmentally regulated biosynthetic pathways 987:1033 developmentally regulated biosynthetic pathways 987:1033 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 3 53 theme complex 274:280 arg1 regions 247:253 the interacting regions 231:253 the interacting regions of the Notch1-DLL4 complex 231:280 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 6 54 theme biosynthetic 1013:1024 arg1 pathways 1026:1033 developmentally regulated biosynthetic pathways 987:1033 developmentally regulated biosynthetic pathways 987:1033 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 5 55 from residues 734:741 arg1 DLL4 746:749 DLL4 746:749 DLL4 746:749 Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. 25700513 1 56 theme Notch1 41:46 arg1 engagement 48:57 Notch1 engagement 41:57 Notch1 engagement of Delta-like 4 41:73 Structural basis for Notch1 engagement of Delta-like 4. 25700513 6 57 theme regulated 1003:1011 arg1 pathways 1026:1033 developmentally regulated biosynthetic pathways 987:1033 developmentally regulated biosynthetic pathways 987:1033 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 25700513 3 58 theme assisted 335:342 arg1 orientation 323:333 a two-site, antiparallel binding orientation 290:333 orientation 323:333 The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. 25700513 6 59 theme sites 924:928 arg1 modifications 886:898 posttranslational modifications 868:898 posttranslational modifications of their ligand binding sites 868:928 The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways. 24820161 0 0 theme enzymatic 75:83 arg1 activity 85:92 enzymatic activity 75:92 enzymatic activity 75:92 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 2 1 gly C-mannosylation 284:298 arg1 glycosylation 269:281 the protein glycosylation 257:281 the protein glycosylation 257:281 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 2 1 gly C-mannosylation 284:298 arg1 ribonuclease 324:335 ribonuclease 2 324:337 ribonuclease 2 324:337 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 5 2 theme mass 709:712 arg1 spectrometry 714:725 mass spectrometry 709:725 mass spectrometry 709:725 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 7 3 theme opposite 1112:1119 arg1 direction 1121:1129 the opposite direction 1108:1129 the opposite direction 1108:1129 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 8 4 theme enzymatic 1467:1475 arg1 activity 1477:1484 enzymatic activity 1467:1484 enzymatic activity 1467:1484 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 2 5 gly C-mannosylated 383:396 arg1 proteins 349:356 some proteins 344:356 some proteins 344:356 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 8 6 theme C-mannosylation 1347:1361 arg1 presence 1335:1342 the presence 1331:1342 the presence of C-mannosylation among HYAL family proteins 1331:1388 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 7 7 from Trp¹³⁰ 1030:1035 arg1 C-mannosylation 1002:1016 C-mannosylation 1002:1016 C-mannosylation of HYAL1 at Trp¹³⁰ 1002:1035 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 7 8 theme site 1082:1085 arg1 conformation 1045:1056 conformation 1045:1056 conformation of the catalytic active site 1045:1085 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 6 9 theme secreted 925:932 arg1 HYAL1 934:938 secreted HYAL1 925:938 secreted HYAL1 925:938 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 1 10 gly glycosylation 103:115 arg1 protein 185:191 many protein functions 180:201 many protein functions 180:201 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 5 11 gly C-mannosylated 778:791 arg2 Trp³²¹ 814:819 Trp³²¹ 814:819 Trp³²¹ 814:819 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 5 11 gly C-mannosylated 778:791 arg2 Trp¹³⁰ 796:801 Trp¹³⁰ 796:801 Trp¹³⁰ 796:801 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 5 11 gly C-mannosylated 778:791 arg1 HYAL1 769:773 intracellular HYAL1 755:773 intracellular HYAL1 755:773 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 2 12 theme protein 261:267 arg1 glycosylation 269:281 the protein glycosylation 257:281 the protein glycosylation 257:281 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 7 13 theme catalytic 1065:1073 arg1 site 1082:1085 the catalytic active site 1061:1085 the catalytic active site 1061:1085 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 3 14 theme hyaluronic 508:517 arg1 acid 519:522 degrading hyaluronic acid 498:522 degrading hyaluronic acid (HA) 498:527 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 14 theme hyaluronic 508:517 arg1 Hyaluronidase 473:485 Hyaluronidase 1 473:487 Hyaluronidase 1 (HYAL1) 473:495 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 14 theme hyaluronic 508:517 arg1 HA 525:526 HA 525:526 HA 525:526 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 6 15 contain possessed 895:903 arg1 it 892:893 it 892:893 it 892:893 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 6 15 contain possessed 895:903 arg2 activity 915:922 enzymatic activity 905:922 enzymatic activity 905:922 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 2 16 theme modifications 423:435 arg1 effects 408:414 effects 408:414 effects of its modifications for target proteins 408:455 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 1 17 theme many 180:183 arg1 secretion 243:251 secretion 243:251 secretion 243:251 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 17 theme many 180:183 arg1 folding 231:237 folding 231:237 folding 231:237 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 17 theme many 180:183 arg1 stability 220:228 protein stability 212:228 protein stability 212:228 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 17 theme many 180:183 arg1 functions 193:201 many protein functions 180:201 many protein functions 180:201 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 18 theme protein 185:191 arg1 secretion 243:251 secretion 243:251 secretion 243:251 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 18 theme protein 185:191 arg1 folding 231:237 folding 231:237 folding 231:237 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 18 theme protein 185:191 arg1 stability 220:228 protein stability 212:228 protein stability 212:228 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 18 theme protein 185:191 arg1 functions 193:201 many protein functions 180:201 many protein functions 180:201 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 0 19 theme hyaluronidase 25:37 arg1 C-mannosylation 0:14 C-mannosylation 0:14 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 0:93 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 6 20 theme enzymatic 905:913 arg1 activity 915:922 enzymatic activity 905:922 enzymatic activity 905:922 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 4 21 from effect 667:672 arg1 HYAL1 696:700 HYAL1 696:700 HYAL1 696:700 In this study, we examined whether HYAL1 is C-mannosylated or not, and the effect of C-mannosylation on HYAL1. 24820161 1 22 theme Protein 95:101 arg1 modifications 148:160 the post-translational modifications 125:160 the post-translational modifications 125:160 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 22 theme Protein 95:101 arg1 one 118:120 one 118:120 one 118:120 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 22 theme Protein 95:101 arg1 glycosylation 103:115 Protein glycosylation 95:115 Protein glycosylation 95:115 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 3 23 theme C-mannosylation 548:562 arg1 sites 564:568 two predicted C-mannosylation sites 534:568 two predicted C-mannosylation sites 534:568 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 0 24 theme human 19:23 arg1 hyaluronidase 25:37 human hyaluronidase 1 19:39 human hyaluronidase 1 19:39 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 3 25 contain has 530:532 arg1 Hyaluronidase 473:485 Hyaluronidase 1 473:487 Hyaluronidase 1 (HYAL1) 473:495 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 25 contain has 530:532 arg1 HYAL1 490:494 HYAL1 490:494 HYAL1 490:494 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 25 contain has 530:532 arg2 sites 564:568 two predicted C-mannosylation sites 534:568 two predicted C-mannosylation sites 534:568 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 25 contain has 530:532 arg1 acid 519:522 degrading hyaluronic acid 498:522 degrading hyaluronic acid (HA) 498:527 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 26 theme degrading 498:506 arg1 acid 519:522 degrading hyaluronic acid 498:522 degrading hyaluronic acid (HA) 498:527 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 26 theme degrading 498:506 arg1 Hyaluronidase 473:485 Hyaluronidase 1 473:487 Hyaluronidase 1 (HYAL1) 473:495 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 3 26 theme degrading 498:506 arg1 HA 525:526 HA 525:526 HA 525:526 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 4 27 theme C-mannosylation 677:691 arg1 effect 667:672 the effect 663:672 the effect of C-mannosylation on HYAL1 663:700 In this study, we examined whether HYAL1 is C-mannosylated or not, and the effect of C-mannosylation on HYAL1. 24820161 3 28 gly C-mannosylation 548:562 arg2 sites 564:568 two predicted C-mannosylation sites 534:568 two predicted C-mannosylation sites 534:568 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 7 29 theme HYAL1 1021:1025 arg1 C-mannosylation 1002:1016 C-mannosylation 1002:1016 C-mannosylation of HYAL1 at Trp¹³⁰ 1002:1035 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 0 30 gly C-mannosylation 0:14 arg1 hyaluronidase 25:37 human hyaluronidase 1 19:39 human hyaluronidase 1 19:39 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 8 31 theme possible 1415:1422 arg1 roles 1424:1428 possible roles 1415:1428 possible roles of C-mannosylation for secretion and enzymatic activity 1415:1484 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 5 32 theme intracellular 755:767 arg1 HYAL1 769:773 intracellular HYAL1 755:773 intracellular HYAL1 755:773 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 7 33 theme Computer 964:971 arg1 simulation 973:982 Computer simulation 964:982 Computer simulation 964:982 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 7 34 theme enzymatic 1252:1260 arg1 activity 1262:1269 its enzymatic activity 1248:1269 its enzymatic activity 1248:1269 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 2 35 theme target 441:446 arg1 proteins 448:455 target proteins 441:455 target proteins 441:455 In the protein glycosylation, C-mannosylation was first identified in ribonuclease 2, and some proteins have been reported to be C-mannosylated; however, effects of its modifications for target proteins remain unclear. 24820161 8 36 theme HYAL 1369:1372 arg1 proteins 1381:1388 HYAL family proteins 1369:1388 HYAL family proteins 1369:1388 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 5 37 from Trp¹³⁰ 796:801 arg1 C-mannosylated 778:791 C-mannosylated 778:791 C-mannosylated 778:791 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 0 38 theme possible 42:49 arg1 roles 51:55 possible roles 42:55 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 0:93 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 3 39 theme predicted 538:546 arg1 sites 564:568 two predicted C-mannosylation sites 534:568 two predicted C-mannosylation sites 534:568 Hyaluronidase 1 (HYAL1), degrading hyaluronic acid (HA), has two predicted C-mannosylation sites at Trp¹³⁰ and Trp³²¹. 24820161 7 40 gly C-mannosylation 1002:1016 arg1 Trp¹³⁰ 1030:1035 Trp¹³⁰ 1030:1035 Trp¹³⁰ 1030:1035 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 7 40 gly C-mannosylation 1002:1016 arg1 HYAL1 1021:1025 HYAL1 1021:1025 HYAL1 1021:1025 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 6 41 theme conditioned 869:879 arg1 medium 881:886 conditioned medium 869:886 conditioned medium 869:886 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 4 42 gly C-mannosylated 636:649 arg1 HYAL1 627:631 HYAL1 627:631 HYAL1 627:631 In this study, we examined whether HYAL1 is C-mannosylated or not, and the effect of C-mannosylation on HYAL1. 24820161 7 43 from Glu¹³¹ 1098:1103 arg1 direction 1121:1129 the opposite direction 1108:1129 the opposite direction 1108:1129 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 1 44 theme protein 212:218 arg1 stability 220:228 protein stability 212:228 protein stability 212:228 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 8 45 theme first 1300:1304 arg1 report 1306:1311 the first report 1296:1311 the first report that demonstrates the presence of C-mannosylation among HYAL family proteins 1296:1388 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 8 45 theme first 1300:1304 arg1 this 1288:1291 this 1288:1291 this 1288:1291 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 8 46 theme family 1374:1379 arg1 proteins 1381:1388 HYAL family proteins 1369:1388 HYAL family proteins 1369:1388 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 7 47 theme active 1075:1080 arg1 site 1082:1085 the catalytic active site 1061:1085 the catalytic active site 1061:1085 Computer simulation demonstrated that C-mannosylation of HYAL1 at Trp¹³⁰ changed conformation of the catalytic active site, and faced Glu¹³¹ in the opposite direction toward its substrate, HA, indicating that C-mannosylation will negatively regulate its secretion, and will attenuate its enzymatic activity. 24820161 8 48 theme C-mannosylation 1433:1447 arg1 roles 1424:1428 possible roles 1415:1428 possible roles of C-mannosylation for secretion and enzymatic activity 1415:1484 Taken together, this is the first report that demonstrates the presence of C-mannosylation among HYAL family proteins, and our results suggest possible roles of C-mannosylation for secretion and enzymatic activity. 24820161 5 49 from Trp³²¹ 814:819 arg1 C-mannosylated 778:791 C-mannosylated 778:791 C-mannosylated 778:791 Using mass spectrometry, we first demonstrated that intracellular HYAL1 is C-mannosylated at Trp¹³⁰ but not at Trp³²¹. 24820161 1 50 theme post-translational 129:146 arg1 modifications 148:160 the post-translational modifications 125:160 the post-translational modifications 125:160 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 0 51 dep C-mannosylation 0:14 arg1 roles 51:55 possible roles 42:55 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 0:93 C-mannosylation of human hyaluronidase 1: possible roles for secretion and enzymatic activity. 24820161 6 52 gly C-mannosylated 948:961 arg1 HYAL1 934:938 secreted HYAL1 925:938 secreted HYAL1 925:938 Surprisingly, although HYAL1 was secreted into conditioned medium and it possessed enzymatic activity, secreted HYAL1 was not C-mannosylated. 24820161 1 53 theme modifications 148:160 arg1 modifications 148:160 the post-translational modifications 125:160 the post-translational modifications 125:160 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 53 theme modifications 148:160 arg1 one 118:120 one 118:120 one 118:120 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 24820161 1 53 theme modifications 148:160 arg1 glycosylation 103:115 Protein glycosylation 95:115 Protein glycosylation 95:115 Protein glycosylation, one of the post-translational modifications, is important for many protein functions, such as protein stability, folding and secretion. 25406038 0 0 from approach 16:23 arg1 C-terminus 81:90 the C-terminus 77:90 the C-terminus of the intestinal MUC2mucin 77:118 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 11 1 gly glycoproteins 1432:1444 arg1 glycoproteins 1432:1444 complex glycoproteins 1424:1444 complex glycoproteins such as mucins 1424:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 1 gly glycoproteins 1432:1444 arg1 mucins 1454:1459 mucins 1454:1459 mucins 1454:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 2 2 dep O-glycosylated 280:293 arg1 binds 359:363 binds 359:363 binds bacteria 359:372 The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. 25406038 5 3 theme MUC2 739:742 arg1 acids 724:728 the last 981 amino acids 705:728 the last 981 amino acids of human MUC2 705:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 5 4 theme CHO-K1 760:765 arg1 cells 767:771 CHO-K1 cells 760:771 CHO-K1 cells 760:771 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 3 5 theme proper 501:506 arg1 folding 516:522 proper protein folding 501:522 proper protein folding 501:522 Glycosylation outside the mucin domain is suggested to be important for proper protein folding and protection against intestinal proteases. 25406038 5 6 theme purified 657:664 arg1 protein 686:692 A purified 250 kDa recombinant protein 655:692 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 655:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 4 7 theme regions 601:607 arg1 glycosylation 578:590 glycosylation 578:590 glycosylation of these regions of the MUC2 578:619 However, glycosylation of these regions of the MUC2 has not been extensively studied. 25406038 2 8 gly O-glycosylated 280:293 arg1 domain 263:268 The protein's central mucin domain 235:268 The protein's central mucin domain 235:268 The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. 25406038 5 9 theme 250 666:668 arg1 kDa 670:672 kDa 670:672 kDa 670:672 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 3 10 theme intestinal 547:556 arg1 proteases 558:566 intestinal proteases 547:566 intestinal proteases 547:566 Glycosylation outside the mucin domain is suggested to be important for proper protein folding and protection against intestinal proteases. 25406038 0 11 theme MUC2mucin 110:118 arg1 C-terminus 81:90 the C-terminus 77:90 the C-terminus of the intestinal MUC2mucin 77:118 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 8 12 theme multiple 1011:1018 arg1 approach 1027:1034 The multiple enzyme approach 1007:1034 The multiple enzyme approach 1007:1034 The multiple enzyme approach increased peptide coverage from 36% when only using trypsin, to 86%. 25406038 9 13 theme N-glycan 1125:1132 arg1 sites 1144:1148 the 18 N-glycan consensus sites 1118:1148 the 18 N-glycan consensus sites 1118:1148 Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. 25406038 11 14 gly glycosylation 1407:1419 arg1 glycoproteins 1432:1444 complex glycoproteins 1424:1444 complex glycoproteins such as mucins 1424:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 14 gly glycosylation 1407:1419 arg1 mucins 1454:1459 mucins 1454:1459 mucins 1454:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 1 15 theme structural 167:176 arg1 component 178:186 the main structural component 158:186 the main structural component of the mucus that covers the colon epithelium 158:232 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 0 16 theme intestinal 99:108 arg1 MUC2mucin 110:118 the intestinal MUC2mucin 95:118 the intestinal MUC2mucin 95:118 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 4 17 gly glycosylation 578:590 arg1 regions 601:607 these regions 595:607 these regions of the MUC2 595:619 However, glycosylation of these regions of the MUC2 has not been extensively studied. 25406038 11 18 theme presented 1298:1306 arg1 method 1308:1313 The presented method 1294:1313 The presented method of protein digestion 1294:1334 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 5 19 theme human 733:737 arg1 MUC2 739:742 human MUC2 733:742 human MUC2 733:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 0 20 theme enzyme 9:14 arg1 approach 16:23 Multiple enzyme approach 0:23 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin 0:118 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 9 21 theme sites 1144:1148 arg1 sites 1144:1148 the 18 N-glycan consensus sites 1118:1148 the 18 N-glycan consensus sites 1118:1148 Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. 25406038 9 21 theme sites 1144:1148 arg1 glycosylated 1169:1180 glycosylated 1169:1180 glycosylated 1169:1180 Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. 25406038 9 21 theme sites 1144:1148 arg1 Seventeen 1105:1113 Seventeen 1105:1113 Seventeen 1105:1113 Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. 25406038 0 22 theme Multiple 0:7 arg1 approach 16:23 Multiple enzyme approach 0:23 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin 0:118 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 8 23 theme enzyme 1020:1025 arg1 approach 1027:1034 The multiple enzyme approach 1007:1034 The multiple enzyme approach 1007:1034 The multiple enzyme approach increased peptide coverage from 36% when only using trypsin, to 86%. 25406038 6 24 dep analyzed 790:797 arg1 followed 836:843 followed 836:843 followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N 836:912 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 8 25 theme peptide 1046:1052 arg1 coverage 1054:1061 peptide coverage 1046:1061 peptide coverage 1046:1061 The multiple enzyme approach increased peptide coverage from 36% when only using trypsin, to 86%. 25406038 11 26 theme complex 1424:1430 arg1 glycoproteins 1432:1444 complex glycoproteins 1424:1444 complex glycoproteins such as mucins 1424:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 26 theme complex 1424:1430 arg1 mucins 1454:1459 mucins 1454:1459 mucins 1454:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 2 27 theme epithelial 411:420 arg1 cells 422:426 the epithelial cells 407:426 the epithelial cells 407:426 The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. 25406038 5 28 theme last 709:712 arg1 acids 724:728 the last 981 amino acids 705:728 the last 981 amino acids of human MUC2 705:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 6 29 theme in-gel 848:853 arg1 digestion 855:863 in-gel digestion 848:863 in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N 848:912 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 1 30 theme main 162:165 arg1 component 178:186 the main structural component 158:186 the main structural component of the mucus that covers the colon epithelium 158:232 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 11 31 theme protein 1318:1324 arg1 digestion 1326:1334 protein digestion 1318:1334 protein digestion 1318:1334 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 10 32 gly O-glycopeptides 1244:1258 arg2 O-glycopeptides 1244:1258 14 O-glycopeptides 1241:1258 14 O-glycopeptides 1241:1258 Fifty-six N-glycopeptides covering 10 N-glycan sites, and 14 O-glycopeptides were sequenced and characterized. 25406038 11 33 theme glycosylation 1407:1419 arg1 complexity 1393:1402 complexity 1393:1402 complexity 1393:1402 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 33 theme glycosylation 1407:1419 arg1 density 1381:1387 density 1381:1387 density 1381:1387 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 34 theme digestion 1326:1334 arg1 method 1308:1313 The presented method 1294:1313 The presented method of protein digestion 1294:1334 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 5 35 contain containing 694:703 arg1 protein 686:692 A purified 250 kDa recombinant protein 655:692 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 655:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 5 35 contain containing 694:703 arg2 acids 724:728 the last 981 amino acids 705:728 the last 981 amino acids of human MUC2 705:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 5 36 theme amino 718:722 arg1 acids 724:728 the last 981 amino acids 705:728 the last 981 amino acids of human MUC2 705:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 11 37 theme better 1356:1361 arg1 insights 1363:1370 better insights 1356:1370 better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins 1356:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 7 38 theme ETD 979:981 arg1 combination 959:969 a combination 957:969 a combination of CID, ETD, and HCD fragmentation 957:1004 Peptides were analyzed by nLC/MS/MS using a combination of CID, ETD, and HCD fragmentation. 25406038 3 39 theme mucin 455:459 arg1 domain 461:466 the mucin domain 451:466 the mucin domain 451:466 Glycosylation outside the mucin domain is suggested to be important for proper protein folding and protection against intestinal proteases. 25406038 0 40 theme glycan 53:58 arg1 modifications 60:72 glycan modifications 53:72 glycan modifications 53:72 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 5 41 theme kDa 670:672 arg1 protein 686:692 A purified 250 kDa recombinant protein 655:692 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 655:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 11 42 used used 1343:1346 arg2 method 1308:1313 The presented method 1294:1313 The presented method of protein digestion 1294:1334 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 11 43 dep density 1381:1387 arg1 the 1377:1379 the 1377:1379 the 1377:1379 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 9 44 theme consensus 1134:1142 arg1 sites 1144:1148 the 18 N-glycan consensus sites 1118:1148 the 18 N-glycan consensus sites 1118:1148 Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. 25406038 5 45 theme recombinant 674:684 arg1 protein 686:692 A purified 250 kDa recombinant protein 655:692 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 655:742 A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. 25406038 7 46 theme CID 974:976 arg1 combination 959:969 a combination 957:969 a combination of CID, ETD, and HCD fragmentation 957:1004 Peptides were analyzed by nLC/MS/MS using a combination of CID, ETD, and HCD fragmentation. 25406038 10 47 gly N-glycopeptides 1193:1207 arg2 N-glycopeptides 1193:1207 Fifty-six N-glycopeptides 1183:1207 Fifty-six N-glycopeptides covering 10 N-glycan sites 1183:1234 Fifty-six N-glycopeptides covering 10 N-glycan sites, and 14 O-glycopeptides were sequenced and characterized. 25406038 10 48 theme Fifty-six 1183:1191 arg1 N-glycopeptides 1193:1207 Fifty-six N-glycopeptides 1183:1207 Fifty-six N-glycopeptides covering 10 N-glycan sites 1183:1234 Fifty-six N-glycopeptides covering 10 N-glycan sites, and 14 O-glycopeptides were sequenced and characterized. 25406038 6 49 theme F 823:823 arg1 treatment 825:833 PNGase F treatment 816:833 PNGase F treatment 816:833 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 2 50 theme mucin 257:261 arg1 domain 263:268 The protein's central mucin domain 235:268 The protein's central mucin domain 235:268 The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. 25406038 1 51 theme polymeric 125:133 arg1 MUC2 141:144 The polymeric mucin MUC2 121:144 The polymeric mucin MUC2 121:144 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 1 52 theme colon 217:221 arg1 epithelium 223:232 the colon epithelium 213:232 the colon epithelium 213:232 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 7 53 theme HCD 988:990 arg1 fragmentation 992:1004 HCD fragmentation 988:1004 HCD fragmentation 988:1004 Peptides were analyzed by nLC/MS/MS using a combination of CID, ETD, and HCD fragmentation. 25406038 6 54 theme PNGase 816:821 arg1 treatment 825:833 PNGase F treatment 816:833 PNGase F treatment 816:833 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 2 55 theme central 249:255 arg1 domain 263:268 The protein's central mucin domain 235:268 The protein's central mucin domain 235:268 The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. 25406038 1 56 theme mucin 135:139 arg1 MUC2 141:144 The polymeric mucin MUC2 121:144 The polymeric mucin MUC2 121:144 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 11 57 theme glycoproteins 1432:1444 arg1 glycosylation 1407:1419 glycosylation 1407:1419 glycosylation of complex glycoproteins such as mucins 1407:1459 The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins. 25406038 7 58 theme fragmentation 992:1004 arg1 combination 959:969 a combination 957:969 a combination of CID, ETD, and HCD fragmentation 957:1004 Peptides were analyzed by nLC/MS/MS using a combination of CID, ETD, and HCD fragmentation. 25406038 1 59 theme mucus 195:199 arg1 component 178:186 the main structural component 158:186 the main structural component of the mucus that covers the colon epithelium 158:232 The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. 25406038 3 60 theme protein 508:514 arg1 folding 516:522 proper protein folding 501:522 proper protein folding 501:522 Glycosylation outside the mucin domain is suggested to be important for proper protein folding and protection against intestinal proteases. 25406038 0 61 theme modifications 60:72 arg1 characterization 33:48 the characterization 29:48 the characterization of glycan modifications 29:72 Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin. 25406038 6 62 with digestion 855:863 arg1 trypsin 870:876 trypsin 870:876 trypsin 870:876 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 6 62 with digestion 855:863 arg1 subtilisin 893:902 subtilisin 893:902 subtilisin 893:902 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 6 62 with digestion 855:863 arg1 Asp-N 908:912 Asp-N 908:912 Asp-N 908:912 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 6 62 with digestion 855:863 arg1 chymotrypsin 879:890 chymotrypsin 879:890 chymotrypsin 879:890 The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. 25406038 10 63 theme N-glycan 1221:1228 arg1 sites 1230:1234 10 N-glycan sites 1218:1234 10 N-glycan sites 1218:1234 Fifty-six N-glycopeptides covering 10 N-glycan sites, and 14 O-glycopeptides were sequenced and characterized. 25406038 4 64 theme MUC2 616:619 arg1 regions 601:607 these regions 595:607 these regions of the MUC2 595:619 However, glycosylation of these regions of the MUC2 has not been extensively studied. 25227423 0 0 theme antibodies 77:86 arg1 binding 61:67 binding 61:67 binding of host antibodies 61:86 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 1 1 theme different 301:309 arg1 assumptions 311:321 widely different assumptions 294:321 widely different assumptions 294:321 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 4 2 from analysis 733:740 arg1 vicinity 762:769 the vicinity 758:769 the vicinity of N-glycans 758:782 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 1 3 theme role 133:136 arg1 extent 119:124 The extent 115:124 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies 115:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 0 4 theme host 72:75 arg1 antibodies 77:86 host antibodies 72:86 host antibodies 72:86 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 0 5 from glycans 9:15 arg1 hemagglutinin 37:49 influenza A H3N2 hemagglutinin 20:49 influenza A H3N2 hemagglutinin 20:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 4 6 theme better 807:812 arg1 understanding 814:826 a better understanding 805:826 a better understanding of their protective role 805:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 1 7 from role 133:136 arg1 HA 210:211 HA 210:211 HA 210:211 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 7 from role 133:136 arg1 hemagglutinin 195:207 shielding influenza A hemagglutinin 173:207 shielding influenza A hemagglutinin (HA) against host antibodies 173:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 5 8 theme sequences 914:922 arg1 analysis 875:882 an analysis 872:882 an analysis of over 6000 influenza A H3N2 sequences 872:922 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 3 9 theme responses 653:661 arg1 antibodies 558:567 antibodies 558:567 antibodies that bind close to N-glycans 558:596 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 3 9 theme responses 653:661 arg1 feature 624:630 a rare or commonplace feature 602:630 a rare or commonplace feature of human herd immune responses to influenza AHA 602:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 5 10 theme influenza 897:905 arg1 sequences 914:922 over 6000 influenza A H3N2 sequences 887:922 over 6000 influenza A H3N2 sequences 887:922 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 4 11 theme paper 697:701 arg1 aim 685:687 The aim 681:687 The aim of this paper 681:701 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 1 12 theme host 222:225 arg1 antibodies 227:236 host antibodies 222:236 host antibodies 222:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 3 13 theme immune 646:651 arg1 responses 653:661 human herd immune responses 635:661 human herd immune responses to influenza AHA 635:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 5 14 theme A 907:907 arg1 sequences 914:922 over 6000 influenza A H3N2 sequences 887:922 over 6000 influenza A H3N2 sequences 887:922 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 2 15 theme common 328:333 arg1 assumption 335:344 One common assumption 324:344 One common assumption 324:344 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 5 16 theme antigenic 1018:1026 arg1 escape 1028:1033 antigenic escape 1018:1033 antigenic escape from host antibodies 1018:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 0 17 link N-linked 0:7 arg1 glycans 9:15 N-linked glycans 0:15 N-linked glycans on influenza A H3N2 hemagglutinin 0:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 4 18 theme computational 719:731 arg1 analysis 733:740 a computational analysis 717:740 a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role 717:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 1 19 theme N-linked 141:148 arg1 N-glycans 159:167 N-glycans 159:167 N-glycans 159:167 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 19 theme N-linked 141:148 arg1 glycans 150:156 N-linked glycans 141:156 N-linked glycans (N-glycans) 141:168 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 2 20 theme glycosylation 416:428 arg1 residues 390:397 surface residues 382:397 surface residues that are near to glycosylation sites 382:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 2 20 theme glycosylation 416:428 arg1 sites 430:434 glycosylation sites 416:434 glycosylation sites 416:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 1 21 theme glycans 150:156 arg1 role 133:136 the role 129:136 the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies 129:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 6 22 gly N-glycosylation 1107:1121 arg2 site 1123:1126 an N-glycosylation site 1104:1126 an N-glycosylation site 1104:1126 Fifteen of these residues occur within 10 Å of an N-glycosylation site. 25227423 1 23 link N-linked 141:148 arg1 N-glycans 159:167 N-glycans 159:167 N-glycans 159:167 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 23 link N-linked 141:148 arg1 glycans 150:156 N-linked glycans 141:156 N-linked glycans (N-glycans) 141:168 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 24 from extent 119:124 arg1 HA 210:211 HA 210:211 HA 210:211 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 24 from extent 119:124 arg1 hemagglutinin 195:207 shielding influenza A hemagglutinin 173:207 shielding influenza A hemagglutinin (HA) against host antibodies 173:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 7 25 theme host 1320:1323 arg1 antibodies 1325:1334 host antibodies 1320:1334 host antibodies 1320:1334 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 7 26 theme N-glycan 1354:1361 arg1 site 1374:1377 an N-glycan attachment site 1351:1377 an N-glycan attachment site 1351:1377 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 0 27 theme N-linked 0:7 arg1 glycans 9:15 N-linked glycans 0:15 N-linked glycans on influenza A H3N2 hemagglutinin 0:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 7 28 theme antibodies 1325:1334 arg1 inability 1307:1315 the inability 1303:1315 the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues 1303:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 6 29 theme residues 1074:1081 arg1 residues 1074:1081 these residues 1068:1081 these residues 1068:1081 Fifteen of these residues occur within 10 Å of an N-glycosylation site. 25227423 6 29 theme residues 1074:1081 arg1 Fifteen 1057:1063 Fifteen 1057:1063 Fifteen 1057:1063 Fifteen of these residues occur within 10 Å of an N-glycosylation site. 25227423 7 30 theme residues 1431:1438 arg1 masking 1408:1414 the physical masking 1395:1414 the physical masking of neighboring residues 1395:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 0 31 theme influenza 20:28 arg1 hemagglutinin 37:49 influenza A H3N2 hemagglutinin 20:49 influenza A H3N2 hemagglutinin 20:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 7 32 theme surface 1239:1245 arg1 ofHA 1247:1250 the surface ofHA 1235:1250 the surface ofHA 1235:1250 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 7 33 theme shielding 1262:1270 arg1 effect 1272:1277 any shielding effect 1258:1277 any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues 1258:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 7 34 theme attributable 1287:1298 arg1 effect 1272:1277 any shielding effect 1258:1277 any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues 1258:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 7 35 theme close 1203:1207 arg1 proximity 1209:1217 close proximity 1203:1217 close proximity to N-glycans on the surface ofHA 1203:1250 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 5 36 theme H3N2 909:912 arg1 sequences 914:922 over 6000 influenza A H3N2 sequences 887:922 over 6000 influenza A H3N2 sequences 887:922 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 7 37 theme neighboring 1419:1429 arg1 residues 1431:1438 neighboring residues 1419:1438 neighboring residues 1419:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 3 38 theme experimental 527:538 arg1 evidence 540:547 experimental evidence 527:547 experimental evidence 527:547 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 0 39 theme H3N2 32:35 arg1 hemagglutinin 37:49 influenza A H3N2 hemagglutinin 20:49 influenza A H3N2 hemagglutinin 20:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 3 40 theme commonplace 612:622 arg1 antibodies 558:567 antibodies 558:567 antibodies that bind close to N-glycans 558:596 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 3 40 theme commonplace 612:622 arg1 feature 624:630 a rare or commonplace feature 602:630 a rare or commonplace feature of human herd immune responses to influenza AHA 602:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 5 41 theme residues 934:941 arg1 set 927:929 a set 925:929 a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies 925:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 5 41 theme residues 934:941 arg1 residues 934:941 residues 934:941 residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies 934:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 7 42 theme physical 1399:1406 arg1 masking 1408:1414 the physical masking 1395:1414 the physical masking of neighboring residues 1395:1438 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 1 43 theme shielding 173:181 arg1 HA 210:211 HA 210:211 HA 210:211 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 43 theme shielding 173:181 arg1 hemagglutinin 195:207 shielding influenza A hemagglutinin 173:207 shielding influenza A hemagglutinin (HA) against host antibodies 173:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 0 44 theme A 30:30 arg1 hemagglutinin 37:49 influenza A H3N2 hemagglutinin 20:49 influenza A H3N2 hemagglutinin 20:49 N-linked glycans on influenza A H3N2 hemagglutinin constrain binding of host antibodies, but shielding is limited. 25227423 7 45 theme attachment 1363:1372 arg1 site 1374:1377 an N-glycan attachment site 1351:1377 an N-glycan attachment site 1351:1377 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 5 46 gly N-glycosylation 955:969 arg2 sites 971:975 N-glycosylation sites 955:975 N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies 955:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 1 47 theme influenza 183:191 arg1 HA 210:211 HA 210:211 HA 210:211 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 47 theme influenza 183:191 arg1 hemagglutinin 195:207 shielding influenza A hemagglutinin 173:207 shielding influenza A hemagglutinin (HA) against host antibodies 173:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 48 theme different 269:277 arg1 authors 279:285 different authors 269:285 different authors making widely different assumptions 269:321 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 4 49 theme role 848:851 arg1 understanding 814:826 a better understanding 805:826 a better understanding of their protective role 805:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 3 50 theme rare 604:607 arg1 antibodies 558:567 antibodies 558:567 antibodies that bind close to N-glycans 558:596 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 3 50 theme rare 604:607 arg1 feature 624:630 a rare or commonplace feature 602:630 a rare or commonplace feature of human herd immune responses to influenza AHA 602:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 4 51 theme N-glycans 774:782 arg1 vicinity 762:769 the vicinity 758:769 the vicinity of N-glycans 758:782 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 5 52 from antibodies 1045:1054 arg1 escape 1028:1033 antigenic escape 1018:1033 antigenic escape from host antibodies 1018:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 4 53 theme protective 837:846 arg1 role 848:851 their protective role 831:851 their protective role 831:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 3 54 theme human 635:639 arg1 responses 653:661 human herd immune responses 635:661 human herd immune responses to influenza AHA 635:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 5 55 theme N-glycosylation 955:969 arg1 sites 971:975 N-glycosylation sites 955:975 N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies 955:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 4 56 from vicinity 762:769 arg1 analysis 733:740 a computational analysis 717:740 a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role 717:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 3 57 theme herd 641:644 arg1 responses 653:661 human herd immune responses 635:661 human herd immune responses to influenza AHA 635:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25227423 2 58 gly glycosylation 416:428 arg2 residues 390:397 surface residues 382:397 surface residues that are near to glycosylation sites 382:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 2 58 gly glycosylation 416:428 arg2 sites 430:434 glycosylation sites 416:434 glycosylation sites 416:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 5 59 theme adjacent 943:950 arg1 residues 934:941 residues 934:941 residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies 934:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 7 60 from proximity 1209:1217 arg1 ofHA 1247:1250 the surface ofHA 1235:1250 the surface ofHA 1235:1250 Hence, we conclude that it is relatively common for antibodies to bind in close proximity to N-glycans on the surface ofHA, with any shielding effect largely attributable to the inability of host antibodies to bind across an N-glycan attachment site, rather than to the physical masking of neighboring residues. 25227423 1 61 theme A 193:193 arg1 HA 210:211 HA 210:211 HA 210:211 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 61 theme A 193:193 arg1 hemagglutinin 195:207 shielding influenza A hemagglutinin 173:207 shielding influenza A hemagglutinin (HA) against host antibodies 173:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 1 62 from hemagglutinin 195:207 arg1 extent 119:124 The extent 115:124 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies 115:236 The extent of the role of N-linked glycans (N-glycans) in shielding influenza A hemagglutinin (HA) against host antibodies has proved controversial, with different authors making widely different assumptions. 25227423 6 63 theme site 1123:1126 arg1 Å 1099:1099 10 Å 1096:1099 10 Å of an N-glycosylation site 1096:1126 Fifteen of these residues occur within 10 Å of an N-glycosylation site. 25227423 2 64 theme surface 382:388 arg1 residues 390:397 surface residues 382:397 surface residues that are near to glycosylation sites 382:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 2 64 theme surface 382:388 arg1 sites 430:434 glycosylation sites 416:434 glycosylation sites 416:434 One common assumption is that N-glycans physically shield surface residues that are near to glycosylation sites, thereby preventing antibodies from binding to them. 25227423 6 65 theme N-glycosylation 1107:1121 arg1 site 1123:1126 an N-glycosylation site 1104:1126 an N-glycosylation site 1104:1126 Fifteen of these residues occur within 10 Å of an N-glycosylation site. 25227423 4 66 from mutations 745:753 arg1 vicinity 762:769 the vicinity 758:769 the vicinity of N-glycans 758:782 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 5 67 theme host 1040:1043 arg1 antibodies 1045:1054 host antibodies 1040:1054 host antibodies 1040:1054 We identify, from an analysis of over 6000 influenza A H3N2 sequences, a set of residues adjacent to N-glycosylation sites that are highly likely to be involved in antigenic escape from host antibodies. 25227423 4 68 theme mutations 745:753 arg1 analysis 733:740 a computational analysis 717:740 a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role 717:851 The aim of this paper is to present a computational analysis of mutations in the vicinity of N-glycans that will facilitate a better understanding of their protective role. 25227423 3 69 theme influenza 666:674 arg1 AHA 676:678 influenza AHA 666:678 influenza AHA 666:678 However, it is unclear, from existing experimental evidence, whether antibodies that bind close to N-glycans are a rare or commonplace feature of human herd immune responses to influenza AHA. 25155433 5 0 theme GalNAc-T 769:776 arg1 isoenzymes 778:787 The GalNAc-T isoenzymes 765:787 The GalNAc-T isoenzymes 765:787 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 6 1 theme peptide 986:992 arg1 specificities 1004:1016 acceptor peptide substrate specificities 977:1016 acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 977:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 1 2 theme abundant 176:183 arg1 modification 204:215 an abundant and highly diverse modification 173:215 an abundant and highly diverse modification of proteins 173:227 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 1 2 theme abundant 176:183 arg1 O-glycosylation 154:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 5 3 theme isoforms 938:945 arg1 subset 928:933 the specific subset 915:933 the specific subset of isoforms present 915:953 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 5 4 theme specific 919:926 arg1 subset 928:933 the specific subset 915:933 the specific subset of isoforms present 915:953 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 4 5 theme site 582:585 arg1 occupancy 587:595 O-glycan site occupancy 573:595 O-glycan site occupancy 573:595 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 7 6 theme 195 1365:1367 arg1 substrates 1377:1386 195 peptide substrates 1365:1386 195 peptide substrates covering known O-glycosylation sites 1365:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 6 7 theme O-glycosylation 1205:1219 arg1 sites 1221:1225 actual O-glycosylation sites 1198:1225 actual O-glycosylation sites in proteins and number of substrate testable 1198:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 3 8 theme O-glycan 536:543 arg1 structures 545:554 complex O-glycan structures 528:554 complex O-glycan structures 528:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 4 9 theme key 749:751 arg1 GalNAc-Ts 726:734 GalNAc-Ts 726:734 GalNAc-Ts in cells 726:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 9 theme key 749:751 arg1 parameters 753:762 key parameters 749:762 key parameters 749:762 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 9 theme key 749:751 arg1 specificities 665:677 the substrate specificities 651:677 the substrate specificities of individual isoenzymes 651:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 9 theme key 749:751 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 9 theme key 749:751 arg1 repertoire 712:721 the repertoire 708:721 the repertoire of GalNAc-Ts in cells 708:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 6 10 theme recombinant 1024:1034 arg1 GalNAc-Ts 1046:1054 recombinant expressed GalNAc-Ts 1024:1054 recombinant expressed GalNAc-Ts 1024:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 11 theme actual 1198:1203 arg1 sites 1221:1225 actual O-glycosylation sites 1198:1225 actual O-glycosylation sites in proteins and number of substrate testable 1198:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 7 12 theme human 1334:1338 arg1 isoenzymes 1349:1358 10 human GalNAc-T isoenzymes 1331:1358 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites 1331:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 3 13 theme structures 545:554 arg1 structures 545:554 complex O-glycan structures 528:554 complex O-glycan structures 528:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 3 13 theme structures 545:554 arg1 variety 517:523 a variety 515:523 a variety of complex O-glycan structures 515:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 6 14 from sites 1221:1225 arg1 number 1243:1248 number 1243:1248 number of substrate testable 1243:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 14 from sites 1221:1225 arg1 proteins 1230:1237 proteins 1230:1237 proteins 1230:1237 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 2 15 dep 20 312:313 arg1 to 309:310 to 309:310 to 309:310 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 6 16 theme isoforms 1123:1130 arg1 activities 1098:1107 activities 1098:1107 activities of individual isoforms 1098:1130 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 2 17 theme large 290:294 arg1 family 296:301 a large family 288:301 a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues 288:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 4 18 from specificities 665:677 arg1 cells 739:743 cells 739:743 cells 739:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 19 theme substrate 655:663 arg1 GalNAc-Ts 726:734 GalNAc-Ts 726:734 GalNAc-Ts in cells 726:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 19 theme substrate 655:663 arg1 parameters 753:762 key parameters 749:762 key parameters 749:762 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 19 theme substrate 655:663 arg1 specificities 665:677 the substrate specificities 651:677 the substrate specificities of individual isoenzymes 651:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 19 theme substrate 655:663 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 19 theme substrate 655:663 arg1 repertoire 712:721 the repertoire 708:721 the repertoire of GalNAc-Ts in cells 708:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 3 20 theme complex 528:534 arg1 structures 545:554 complex O-glycan structures 528:554 complex O-glycan structures 528:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 6 21 from proteins 1230:1237 arg1 validation 1184:1193 biological validation 1173:1193 biological validation of actual O-glycosylation sites in proteins and number of substrate testable 1173:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 1 22 theme diverse 196:202 arg1 modification 204:215 an abundant and highly diverse modification 173:215 an abundant and highly diverse modification of proteins 173:227 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 1 22 theme diverse 196:202 arg1 O-glycosylation 154:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 0 23 theme polypeptide 8:18 arg1 specificities 57:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 6 24 theme acceptor 977:984 arg1 specificities 1004:1016 acceptor peptide substrate specificities 977:1016 acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 977:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 7 25 theme systematic 1292:1301 arg1 analysis 1303:1310 a systematic analysis 1290:1310 a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites 1290:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 2 26 theme homologous 315:324 arg1 isoenzymes 347:356 up to 20 homologous polypeptide GalNAc-T isoenzymes 306:356 up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues 306:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 3 27 theme GalNAc 424:429 arg1 residues 431:438 These GalNAc residues 418:438 These GalNAc residues 418:438 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 6 28 theme specificities 1004:1016 arg1 analysis 965:972 In vitro analysis 956:972 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 956:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 28 theme specificities 1004:1016 arg1 method 1069:1074 the method 1065:1074 the method of choice for probing activities of individual isoforms 1065:1130 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 4 29 theme GalNAc-Ts 726:734 arg1 GalNAc-Ts 726:734 GalNAc-Ts 726:734 GalNAc-Ts in cells 726:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 29 theme GalNAc-Ts 726:734 arg1 parameters 753:762 key parameters 749:762 key parameters 749:762 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 29 theme GalNAc-Ts 726:734 arg1 specificities 665:677 the substrate specificities 651:677 the substrate specificities of individual isoenzymes 651:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 29 theme GalNAc-Ts 726:734 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 29 theme GalNAc-Ts 726:734 arg1 repertoire 712:721 the repertoire 708:721 the repertoire of GalNAc-Ts in cells 708:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 5 30 theme unique 878:883 arg1 O-glycoproteomes 885:900 unique O-glycoproteomes 878:900 unique O-glycoproteomes dependent on the specific subset of isoforms present 878:953 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 6 31 from validation 1184:1193 arg1 number 1243:1248 number 1243:1248 number of substrate testable 1243:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 31 from validation 1184:1193 arg1 proteins 1230:1237 proteins 1230:1237 proteins 1230:1237 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 5 32 theme dependent 902:910 arg1 O-glycoproteomes 885:900 unique O-glycoproteomes 878:900 unique O-glycoproteomes dependent on the specific subset of isoforms present 878:953 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 0 33 theme isoform 39:45 arg1 specificities 57:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 6 34 theme substrate 994:1002 arg1 specificities 1004:1016 acceptor peptide substrate specificities 977:1016 acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 977:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 4 35 from cells 739:743 arg1 GalNAc-Ts 726:734 GalNAc-Ts 726:734 GalNAc-Ts in cells 726:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 35 from cells 739:743 arg1 parameters 753:762 key parameters 749:762 key parameters 749:762 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 35 from cells 739:743 arg1 specificities 665:677 the substrate specificities 651:677 the substrate specificities of individual isoenzymes 651:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 35 from cells 739:743 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 35 from cells 739:743 arg1 repertoire 712:721 the repertoire 708:721 the repertoire of GalNAc-Ts in cells 708:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 7 36 theme comprehensive 1439:1451 arg1 dataset 1453:1459 a comprehensive dataset 1437:1459 a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome 1437:1528 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 6 37 theme individual 1112:1121 arg1 isoforms 1123:1130 individual isoforms 1112:1130 individual isoforms 1112:1130 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 1 38 theme proteins 220:227 arg1 modification 204:215 an abundant and highly diverse modification 173:215 an abundant and highly diverse modification of proteins 173:227 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 1 38 theme proteins 220:227 arg1 O-glycosylation 154:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 0 39 theme GalNAc-transferase 20:37 arg1 specificities 57:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 4 40 theme O-glycan 573:580 arg1 occupancy 587:595 O-glycan site occupancy 573:595 O-glycan site occupancy 573:595 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 6 41 theme sites 1221:1225 arg1 validation 1184:1193 biological validation 1173:1193 biological validation of actual O-glycosylation sites in proteins and number of substrate testable 1173:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 1 42 theme -type 135:139 arg1 modification 204:215 an abundant and highly diverse modification 173:215 an abundant and highly diverse modification of proteins 173:227 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 1 42 theme -type 135:139 arg1 O-glycosylation 154:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 7 43 theme GalNAc-T 1340:1347 arg1 isoenzymes 1349:1358 10 human GalNAc-T isoenzymes 1331:1358 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites 1331:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 2 44 theme isoenzymes 347:356 arg1 family 296:301 a large family 288:301 a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues 288:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 6 45 dep In 956:957 arg1 vitro 959:963 vitro 959:963 vitro 959:963 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 0 46 dep in 74:75 arg1 vitro 77:81 vitro 77:81 vitro 77:81 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 6 47 theme choice 1079:1084 arg1 analysis 965:972 In vitro analysis 956:972 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 956:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 47 theme choice 1079:1084 arg1 method 1069:1074 the method 1065:1074 the method of choice for probing activities of individual isoforms 1065:1130 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 0 48 theme substrate 47:55 arg1 specificities 57:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 polypeptide GalNAc-transferase isoform substrate specificities 8:69 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 6 49 theme expressed 1036:1044 arg1 GalNAc-Ts 1046:1054 recombinant expressed GalNAc-Ts 1024:1054 recombinant expressed GalNAc-Ts 1024:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 2 50 theme Tyr 404:406 arg1 residues 408:415 Tyr residues 404:415 Tyr residues 404:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 0 51 theme in 74:75 arg1 analysis 83:90 in vitro analysis 74:90 in vitro analysis 74:90 Probing polypeptide GalNAc-transferase isoform substrate specificities by in vitro analysis. 25155433 7 52 with isoenzymes 1349:1358 arg1 substrates 1377:1386 195 peptide substrates 1365:1386 195 peptide substrates covering known O-glycosylation sites 1365:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 2 53 theme O-glycosylation 243:257 arg1 type 235:238 This type 230:238 This type of O-glycosylation 230:257 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 6 54 theme testable 1263:1270 arg1 number 1243:1248 number 1243:1248 number of substrate testable 1243:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 54 theme testable 1263:1270 arg1 proteins 1230:1237 proteins 1230:1237 proteins 1230:1237 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 7 55 theme peptide 1369:1375 arg1 substrates 1377:1386 195 peptide substrates 1365:1386 195 peptide substrates covering known O-glycosylation sites 1365:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 1 56 theme mucin-type 142:151 arg1 modification 204:215 an abundant and highly diverse modification 173:215 an abundant and highly diverse modification of proteins 173:227 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 1 56 theme mucin-type 142:151 arg1 O-glycosylation 154:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation 93:168 N-acetylgalactosaminyltransferase (GalNAc)-type (mucin-type) O-glycosylation is an abundant and highly diverse modification of proteins. 25155433 6 57 theme In 956:957 arg1 analysis 965:972 In vitro analysis 956:972 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts 956:1054 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 57 theme In 956:957 arg1 method 1069:1074 the method 1065:1074 the method of choice for probing activities of individual isoforms 1065:1130 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 7 58 theme known 1397:1401 arg1 sites 1419:1423 known O-glycosylation sites 1397:1423 known O-glycosylation sites 1397:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 7 59 theme isoform-specific 1476:1491 arg1 contributions 1493:1505 isoform-specific contributions 1476:1505 isoform-specific contributions to the O-glycoproteome 1476:1528 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 2 60 theme GalNAc-T 338:345 arg1 isoenzymes 347:356 up to 20 homologous polypeptide GalNAc-T isoenzymes 306:356 up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues 306:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 4 61 from GalNAc-Ts 726:734 arg1 cells 739:743 cells 739:743 cells 739:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 5 62 theme present 947:953 arg1 isoforms 938:945 isoforms 938:945 isoforms present 938:953 The GalNAc-T isoenzymes are differentially expressed in cells and tissues in principle allowing cells to produce unique O-glycoproteomes dependent on the specific subset of isoforms present. 25155433 7 63 theme O-glycosylation 1403:1417 arg1 sites 1419:1423 known O-glycosylation sites 1397:1423 known O-glycosylation sites 1397:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 2 64 theme polypeptide 326:336 arg1 isoenzymes 347:356 up to 20 homologous polypeptide GalNAc-T isoenzymes 306:356 up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues 306:415 This type of O-glycosylation is initiated in the Golgi by a large family of up to 20 homologous polypeptide GalNAc-T isoenzymes that transfer GalNAc to Ser, Thr and possibly Tyr residues. 25155433 7 65 theme isoenzymes 1349:1358 arg1 activity 1319:1326 the activity 1315:1326 the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites 1315:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 3 66 theme large 472:476 arg1 glycosyltransferases 485:504 glycosyltransferases 485:504 glycosyltransferases 485:504 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 3 66 theme large 472:476 arg1 set 478:480 a large set 470:480 a large set of glycosyltransferases to build a variety of complex O-glycan structures 470:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 7 67 gly O-glycosylation 1403:1417 arg2 sites 1419:1423 known O-glycosylation sites 1397:1423 known O-glycosylation sites 1397:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 6 68 from number 1243:1248 arg1 validation 1184:1193 biological validation 1173:1193 biological validation of actual O-glycosylation sites in proteins and number of substrate testable 1173:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 4 69 theme isoenzymes 693:702 arg1 GalNAc-Ts 726:734 GalNAc-Ts 726:734 GalNAc-Ts in cells 726:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 69 theme isoenzymes 693:702 arg1 parameters 753:762 key parameters 749:762 key parameters 749:762 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 69 theme isoenzymes 693:702 arg1 specificities 665:677 the substrate specificities 651:677 the substrate specificities of individual isoenzymes 651:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 69 theme isoenzymes 693:702 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 69 theme isoenzymes 693:702 arg1 repertoire 712:721 the repertoire 708:721 the repertoire of GalNAc-Ts in cells 708:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 4 70 from repertoire 712:721 arg1 cells 739:743 cells 739:743 cells 739:743 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 6 71 gly O-glycosylation 1205:1219 arg2 sites 1221:1225 actual O-glycosylation sites 1198:1225 actual O-glycosylation sites in proteins and number of substrate testable 1198:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 6 72 theme biological 1173:1182 arg1 validation 1184:1193 biological validation 1173:1193 biological validation of actual O-glycosylation sites in proteins and number of substrate testable 1173:1270 In vitro analysis of acceptor peptide substrate specificities using recombinant expressed GalNAc-Ts has been the method of choice for probing activities of individual isoforms, but these studies have been hampered by biological validation of actual O-glycosylation sites in proteins and number of substrate testable. 25155433 7 73 theme activity 1319:1326 arg1 analysis 1303:1310 a systematic analysis 1290:1310 a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites 1290:1423 Here, we present a systematic analysis of the activity of 10 human GalNAc-T isoenzymes with 195 peptide substrates covering known O-glycosylation sites and provide a comprehensive dataset for evaluating isoform-specific contributions to the O-glycoproteome. 25155433 4 74 theme individual 682:691 arg1 isoenzymes 693:702 individual isoenzymes 682:702 individual isoenzymes 682:702 What determines O-glycan site occupancy is still poorly understood, although it is clear that the substrate specificities of individual isoenzymes and the repertoire of GalNAc-Ts in cells are key parameters. 25155433 3 75 theme glycosyltransferases 485:504 arg1 glycosyltransferases 485:504 glycosyltransferases 485:504 glycosyltransferases 485:504 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 25155433 3 75 theme glycosyltransferases 485:504 arg1 set 478:480 a large set 470:480 a large set of glycosyltransferases to build a variety of complex O-glycan structures 470:554 These GalNAc residues are then further elongated by a large set of glycosyltransferases to build a variety of complex O-glycan structures. 27807235 6 0 theme glycan 1111:1116 arg1 epitopes 1118:1125 the glycan epitopes 1107:1125 the glycan epitopes presented on recombinant vaccine candidates 1107:1169 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 1 1 theme Golgi 264:268 arg1 apparatus 270:278 the Golgi apparatus 260:278 the Golgi apparatus 260:278 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 0 2 theme Molecular 0:8 arg1 Architecture 10:21 Molecular Architecture 0:21 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 0:110 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 1 3 theme viral 180:184 arg1 spike 186:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 4 4 theme gp120 881:885 arg1 monomer 887:893 the matched gp120 monomer 869:893 the matched gp120 monomer 869:893 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 4 5 theme quantitative 693:704 arg1 analysis 720:727 a quantitative site-specific analysis 691:727 a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer 691:893 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 10 6 theme dense 1610:1614 arg1 array 1616:1620 the dense array 1606:1620 the dense array of glycans that coat the surface of the viral molecule 1606:1675 These antibodies have evolved to recognize the dense array of glycans that coat the surface of the viral molecule. 27807235 9 7 theme vaccine 1459:1465 arg1 research 1467:1474 HIV-1 vaccine research 1453:1474 HIV-1 vaccine research 1453:1474 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 11 8 theme spike 1806:1810 arg1 folding 1785:1791 the native folding 1774:1791 the native folding of the viral spike 1774:1810 We show how the structure of these glycans is shaped by steric constraints imposed upon them by the native folding of the viral spike. 27807235 8 9 theme wide 1402:1405 arg1 range 1407:1411 a wide range 1400:1411 a wide range of circulating viruses 1400:1434 For some patients infected with HIV-1, highly potent antibodies have been isolated that can neutralize a wide range of circulating viruses. 27807235 11 10 theme steric 1734:1739 arg1 constraints 1741:1751 steric constraints 1734:1751 steric constraints imposed upon them by the native folding of the viral spike 1734:1810 We show how the structure of these glycans is shaped by steric constraints imposed upon them by the native folding of the viral spike. 27807235 5 11 theme glycan 955:960 arg1 remodeling 962:971 a trimer-associated glycan remodeling 935:971 a trimer-associated glycan remodeling 935:971 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 3 12 theme glycosylation 459:471 arg1 pathway 473:479 the glycosylation pathway 455:479 the glycosylation pathway 455:479 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 5 13 theme detailed 909:916 arg1 map 928:930 a detailed molecular map 907:930 a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch 907:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 0 14 theme HIV-1 76:80 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 1 15 theme glycosylated 161:172 arg1 spike 186:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 4 16 theme spike 788:792 arg1 SOSIP.664 801:809 BG505 SOSIP.664 795:809 BG505 SOSIP.664 795:809 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 4 16 theme spike 788:792 arg1 recombinant 734:744 recombinant 734:744 recombinant 734:744 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 0 17 theme Glycoprotein 91:102 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 9 18 theme spike 1556:1560 arg1 mimics 1536:1541 recombinant mimics 1524:1541 recombinant mimics of the viral spike 1524:1560 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 0 19 from Patch 57:61 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 9 20 with immunization 1506:1517 arg1 mimics 1536:1541 recombinant mimics 1524:1541 recombinant mimics of the viral spike 1524:1560 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 2 21 theme further 385:391 arg1 maturation 393:402 further maturation 385:402 further maturation of its extensive glycosylation 385:433 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 7 22 theme virus 1229:1233 arg1 type 1235:1238 human immunodeficiency virus type 1 1206:1240 human immunodeficiency virus type 1 (HIV-1) 1206:1248 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 22 theme virus 1229:1233 arg1 HIV-1 1243:1247 HIV-1 1243:1247 HIV-1 1243:1247 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 23 theme human 1206:1210 arg1 type 1235:1238 human immunodeficiency virus type 1 1206:1240 human immunodeficiency virus type 1 (HIV-1) 1206:1248 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 23 theme human 1206:1210 arg1 HIV-1 1243:1247 HIV-1 1243:1247 HIV-1 1243:1247 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 12 24 theme candidates 1881:1890 arg1 development 1858:1868 the development 1854:1868 the development of vaccine candidates 1854:1890 This information is important in guiding the development of vaccine candidates. 27807235 6 25 theme design 1081:1086 arg1 feature 1088:1094 a critical design feature 1070:1094 a critical design feature 1070:1094 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 6 25 theme design 1081:1086 arg1 formation 1039:1047 The formation 1035:1047 The formation of native trimers 1035:1065 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 4 26 theme HIV-1 776:780 arg1 spike 788:792 the native HIV-1 viral spike 765:792 the native HIV-1 viral spike 765:792 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 2 27 theme extensive 411:419 arg1 glycosylation 421:433 its extensive glycosylation 407:433 its extensive glycosylation 407:433 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 4 28 dep recombinant 734:744 arg1 trimeric 747:754 trimeric 747:754 trimeric 747:754 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 4 28 dep recombinant 734:744 arg1 mimic 756:760 mimic 756:760 mimic 756:760 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 1 29 theme folded 141:146 arg1 spike 186:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 4 30 theme corresponding 828:840 arg1 pseudotrimer 852:863 the corresponding uncleaved pseudotrimer 824:863 the corresponding uncleaved pseudotrimer 824:863 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 10 31 theme molecule 1668:1675 arg1 surface 1647:1653 the surface 1643:1653 the surface of the viral molecule 1643:1675 These antibodies have evolved to recognize the dense array of glycans that coat the surface of the viral molecule. 27807235 0 32 theme Cleavage-Dependent 30:47 arg1 Patch 57:61 the Cleavage-Dependent Mannose Patch 26:61 the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer 26:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 6 33 theme trimers 1059:1065 arg1 formation 1039:1047 The formation 1035:1047 The formation of native trimers 1035:1065 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 6 33 theme trimers 1059:1065 arg1 feature 1088:1094 a critical design feature 1070:1094 a critical design feature 1070:1094 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 3 34 theme antibodies 663:672 arg1 epitopes 621:628 the epitopes 617:628 the epitopes of numerous broadly neutralizing antibodies 617:672 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 8 35 theme viruses 1428:1434 arg1 range 1407:1411 a wide range 1400:1411 a wide range of circulating viruses 1400:1434 For some patients infected with HIV-1, highly potent antibodies have been isolated that can neutralize a wide range of circulating viruses. 27807235 3 36 theme underprocessed 493:506 arg1 glycans 526:532 underprocessed oligomannose-type glycans 493:532 underprocessed oligomannose-type glycans 493:532 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 0 37 theme Patch 57:61 arg1 Architecture 10:21 Molecular Architecture 0:21 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 0:110 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 6 38 theme vaccine 1152:1158 arg1 candidates 1160:1169 recombinant vaccine candidates 1140:1169 recombinant vaccine candidates 1140:1169 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 5 39 theme mannose 1020:1026 arg1 patch 1028:1032 the native mannose patch 1009:1032 the native mannose patch 1009:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 2 40 theme fusion 362:367 arg1 capable 351:357 capable 351:357 capable 351:357 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 1 41 theme gp160 233:237 arg1 protein 249:255 the gp160 precursor protein 229:255 the gp160 precursor protein 229:255 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 7 42 theme antibody-based 1266:1279 arg1 neutralization 1281:1294 antibody-based neutralization 1266:1294 antibody-based neutralization 1266:1294 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 2 43 theme capable 351:357 arg1 spike 345:349 the spike 341:349 the spike capable of fusion 341:367 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 1 44 theme protein 249:255 arg1 cleavage 217:224 protease cleavage 208:224 protease cleavage of the gp160 precursor protein in the Golgi apparatus 208:278 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 9 45 theme HIV-1 1453:1457 arg1 research 1467:1474 HIV-1 vaccine research 1453:1474 HIV-1 vaccine research 1453:1474 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 1 46 theme HIV-1 174:178 arg1 spike 186:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 1 47 theme spike 186:190 arg1 formation 116:124 The formation 112:124 The formation of a correctly folded and natively glycosylated HIV-1 viral spike 112:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 1 47 theme spike 186:190 arg1 dependent 195:203 dependent 195:203 dependent 195:203 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 4 48 theme matched 873:879 arg1 monomer 887:893 the matched gp120 monomer 869:893 the matched gp120 monomer 869:893 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 4 49 theme uncleaved 842:850 arg1 pseudotrimer 852:863 the corresponding uncleaved pseudotrimer 824:863 the corresponding uncleaved pseudotrimer 824:863 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 2 50 theme compact 300:306 arg1 structure 308:316 a compact structure 298:316 a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation 298:433 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 9 51 theme research 1467:1474 arg1 It 1437:1438 It 1437:1438 It 1437:1438 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 9 51 theme research 1467:1474 arg1 goal 1445:1448 a goal 1443:1448 a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike 1443:1560 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 11 52 theme native 1778:1783 arg1 folding 1785:1791 the native folding 1774:1791 the native folding of the viral spike 1774:1810 We show how the structure of these glycans is shaped by steric constraints imposed upon them by the native folding of the viral spike. 27807235 1 53 theme protease 208:215 arg1 cleavage 217:224 protease cleavage 208:224 protease cleavage of the gp160 precursor protein in the Golgi apparatus 208:278 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 10 54 theme glycans 1625:1631 arg1 array 1616:1620 the dense array 1606:1620 the dense array of glycans that coat the surface of the viral molecule 1606:1675 These antibodies have evolved to recognize the dense array of glycans that coat the surface of the viral molecule. 27807235 1 55 from cleavage 217:224 arg1 apparatus 270:278 the Golgi apparatus 260:278 the Golgi apparatus 260:278 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 4 56 theme recombinant 734:744 arg1 analysis 720:727 a quantitative site-specific analysis 691:727 a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer 691:893 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 11 57 theme viral 1800:1804 arg1 spike 1806:1810 the viral spike 1796:1810 the viral spike 1796:1810 We show how the structure of these glycans is shaped by steric constraints imposed upon them by the native folding of the viral spike. 27807235 1 58 gly glycosylated 161:172 arg1 spike 186:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 a correctly folded and natively glycosylated HIV-1 viral spike 129:190 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 27807235 3 59 theme numerous 633:640 arg1 antibodies 663:672 numerous broadly neutralizing antibodies 633:672 numerous broadly neutralizing antibodies 633:672 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 5 60 theme trimer-associated 937:953 arg1 remodeling 962:971 a trimer-associated glycan remodeling 935:971 a trimer-associated glycan remodeling 935:971 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 6 61 theme native 1052:1057 arg1 trimers 1059:1065 native trimers 1052:1065 native trimers 1052:1065 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 0 62 from Trimer 104:109 arg1 Architecture 10:21 Molecular Architecture 0:21 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 0:110 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 5 63 theme localized 986:994 arg1 patch 1028:1032 the native mannose patch 1009:1032 the native mannose patch 1009:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 5 63 theme localized 986:994 arg1 subdomain 996:1004 a localized subdomain 984:1004 a localized subdomain of the native mannose patch 984:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 0 64 theme Soluble 68:74 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 9 65 theme recombinant 1524:1534 arg1 mimics 1536:1541 recombinant mimics 1524:1541 recombinant mimics of the viral spike 1524:1560 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 4 66 theme site-specific 706:718 arg1 analysis 720:727 a quantitative site-specific analysis 691:727 a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer 691:893 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 3 67 from feature 550:556 arg1 design 571:576 immunogen design 561:576 immunogen design 561:576 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 0 68 theme Envelope 82:89 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 5 69 theme molecular 918:926 arg1 map 928:930 a detailed molecular map 907:930 a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch 907:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 0 70 from Architecture 10:21 arg1 Trimer 104:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 a Soluble HIV-1 Envelope Glycoprotein Trimer 66:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 8 71 theme potent 1343:1348 arg1 antibodies 1350:1359 highly potent antibodies 1336:1359 highly potent antibodies 1336:1359 For some patients infected with HIV-1, highly potent antibodies have been isolated that can neutralize a wide range of circulating viruses. 27807235 7 72 theme type 1235:1238 arg1 spike 1197:1201 The envelope spike 1184:1201 The envelope spike of human immunodeficiency virus type 1 (HIV-1) 1184:1248 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 72 theme type 1235:1238 arg1 target 1255:1260 a target 1253:1260 a target for antibody-based neutralization 1253:1294 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 72 theme type 1235:1238 arg1 IMPORTANCE 1172:1181 IMPORTANCE 1172:1181 IMPORTANCE 1172:1181 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 9 73 theme viral 1550:1554 arg1 spike 1556:1560 the viral spike 1546:1560 the viral spike 1546:1560 It is a goal of HIV-1 vaccine research to elicit these antibodies by immunization with recombinant mimics of the viral spike. 27807235 4 74 theme viral 782:786 arg1 spike 788:792 the native HIV-1 viral spike 765:792 the native HIV-1 viral spike 765:792 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 7 75 theme immunodeficiency 1212:1227 arg1 type 1235:1238 human immunodeficiency virus type 1 1206:1240 human immunodeficiency virus type 1 (HIV-1) 1206:1248 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 75 theme immunodeficiency 1212:1227 arg1 HIV-1 1243:1247 HIV-1 1243:1247 HIV-1 1243:1247 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 12 76 theme vaccine 1873:1879 arg1 candidates 1881:1890 vaccine candidates 1873:1890 vaccine candidates 1873:1890 This information is important in guiding the development of vaccine candidates. 27807235 5 77 theme remodeling 962:971 arg1 map 928:930 a detailed molecular map 907:930 a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch 907:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 7 78 theme envelope 1188:1195 arg1 spike 1197:1201 The envelope spike 1184:1201 The envelope spike of human immunodeficiency virus type 1 (HIV-1) 1184:1248 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 78 theme envelope 1188:1195 arg1 target 1255:1260 a target 1253:1260 a target for antibody-based neutralization 1253:1294 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 7 78 theme envelope 1188:1195 arg1 IMPORTANCE 1172:1181 IMPORTANCE 1172:1181 IMPORTANCE 1172:1181 IMPORTANCE: The envelope spike of human immunodeficiency virus type 1 (HIV-1) is a target for antibody-based neutralization. 27807235 6 79 theme critical 1072:1079 arg1 feature 1088:1094 a critical design feature 1070:1094 a critical design feature 1070:1094 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 6 79 theme critical 1072:1079 arg1 formation 1039:1047 The formation 1035:1047 The formation of native trimers 1035:1065 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 2 80 theme glycosylation 421:433 arg1 maturation 393:402 further maturation 385:402 further maturation of its extensive glycosylation 385:433 Cleavage induces a compact structure which not only renders the spike capable of fusion but also limits further maturation of its extensive glycosylation. 27807235 4 81 theme native 769:774 arg1 spike 788:792 the native HIV-1 viral spike 765:792 the native HIV-1 viral spike 765:792 Here we present a quantitative site-specific analysis of a recombinant, trimeric mimic of the native HIV-1 viral spike (BG505 SOSIP.664) compared to the corresponding uncleaved pseudotrimer and the matched gp120 monomer. 27807235 3 82 theme pathway 473:479 arg1 feature 550:556 an important feature 537:556 an important feature in immunogen design 537:576 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 3 82 theme pathway 473:479 arg1 redirection 440:450 The redirection 436:450 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans 436:532 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 0 83 theme Mannose 49:55 arg1 Patch 57:61 the Cleavage-Dependent Mannose Patch 26:61 the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer 26:109 Molecular Architecture of the Cleavage-Dependent Mannose Patch on a Soluble HIV-1 Envelope Glycoprotein Trimer. 27807235 3 84 theme neutralizing 650:661 arg1 antibodies 663:672 numerous broadly neutralizing antibodies 633:672 numerous broadly neutralizing antibodies 633:672 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 10 85 theme viral 1662:1666 arg1 molecule 1668:1675 the viral molecule 1658:1675 the viral molecule 1658:1675 These antibodies have evolved to recognize the dense array of glycans that coat the surface of the viral molecule. 27807235 11 86 theme glycans 1713:1719 arg1 structure 1694:1702 the structure 1690:1702 the structure of these glycans 1690:1719 We show how the structure of these glycans is shaped by steric constraints imposed upon them by the native folding of the viral spike. 27807235 8 87 theme circulating 1416:1426 arg1 viruses 1428:1434 circulating viruses 1416:1434 circulating viruses 1416:1434 For some patients infected with HIV-1, highly potent antibodies have been isolated that can neutralize a wide range of circulating viruses. 27807235 3 88 theme oligomannose-type 508:524 arg1 glycans 526:532 underprocessed oligomannose-type glycans 493:532 underprocessed oligomannose-type glycans 493:532 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 5 89 theme native 1013:1018 arg1 patch 1028:1032 the native mannose patch 1009:1032 the native mannose patch 1009:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 6 90 theme recombinant 1140:1150 arg1 candidates 1160:1169 recombinant vaccine candidates 1140:1169 recombinant vaccine candidates 1140:1169 The formation of native trimers is a critical design feature in shaping the glycan epitopes presented on recombinant vaccine candidates. 27807235 5 91 theme patch 1028:1032 arg1 patch 1028:1032 the native mannose patch 1009:1032 the native mannose patch 1009:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 5 91 theme patch 1028:1032 arg1 subdomain 996:1004 a localized subdomain 984:1004 a localized subdomain of the native mannose patch 984:1032 We present a detailed molecular map of a trimer-associated glycan remodeling that forms a localized subdomain of the native mannose patch. 27807235 3 92 theme important 540:548 arg1 feature 550:556 an important feature 537:556 an important feature in immunogen design 537:576 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 3 92 theme important 540:548 arg1 redirection 440:450 The redirection 436:450 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans 436:532 The redirection of the glycosylation pathway to preserve underprocessed oligomannose-type glycans is an important feature in immunogen design, as glycans contribute to or influence the epitopes of numerous broadly neutralizing antibodies. 27807235 1 93 theme precursor 239:247 arg1 protein 249:255 the gp160 precursor protein 229:255 the gp160 precursor protein 229:255 The formation of a correctly folded and natively glycosylated HIV-1 viral spike is dependent on protease cleavage of the gp160 precursor protein in the Golgi apparatus. 28445724 1 0 from response 159:166 arg1 Env 131:133 shelter Env 123:133 shelter Env from the humoral immune response 123:166 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 5 1 theme Env-elicited 962:973 arg1 ID50 991:994 ID50 991:994 ID50 991:994 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 5 1 theme Env-elicited 962:973 arg1 neutralization 975:988 Env-elicited neutralization 962:988 Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody 962:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 6 2 theme priming 1190:1196 arg1 trimers 1179:1185 site-selective deglycosylated trimers 1149:1185 site-selective deglycosylated trimers 1149:1185 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 6 2 theme priming 1190:1196 arg1 immunogens 1198:1207 priming immunogens 1190:1207 priming immunogens 1190:1207 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 1 3 theme humoral 144:150 arg1 response 159:166 the humoral immune response 140:166 the humoral immune response 140:166 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 3 4 theme guinea 529:534 arg1 pigs 536:539 guinea pigs 529:539 guinea pigs 529:539 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 2 5 from impact 288:293 arg1 elicitation 319:329 elicitation 319:329 elicitation of antibodies against the CD4 supersite 319:369 Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. 28445724 4 6 dep % 753:753 arg1 90 751:752 90 751:752 90 751:752 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 7 with viruses 733:739 arg1 strains 804:810 over 90% breadth against four-glycan-deleted heterologous strains 746:810 over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 746:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 5 8 theme protein-surface 1036:1050 arg1 area 1052:1055 the protein-surface area 1032:1055 the protein-surface area accessible to antibody 1032:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 4 9 theme little 584:589 arg1 neutralization 591:604 little neutralization 584:604 little neutralization against wild-type viruses 584:630 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 10 theme autologous 722:731 arg1 viruses 733:739 four-glycan-deleted autologous viruses 702:739 four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 702:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 11 theme potent 636:641 arg1 neutralization 657:670 potent CD4-supersite neutralization 636:670 potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character) 636:854 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 11 theme potent 636:641 arg1 titers 673:678 titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 673:853 titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 673:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 2 12 theme targeted 248:255 arg1 deglycosylation 257:271 targeted deglycosylation 248:271 targeted deglycosylation 248:271 Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. 28445724 1 13 theme immune 152:157 arg1 response 159:166 the humoral immune response 140:166 the humoral immune response 140:166 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 0 14 theme HIV-1-Glycan 36:47 arg1 Impact 22:27 the Impact 18:27 the Impact of the HIV-1-Glycan 18:47 Quantification of the Impact of the HIV-1-Glycan Shield on Antibody Elicitation. 28445724 3 15 with trimers 398:404 arg1 glycans 418:424 select glycans 411:424 select glycans 411:424 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 5 16 theme surface 933:939 arg1 immunogenicity 887:900 the immunogenicity 883:900 the immunogenicity of the glycan-shielded protein surface 883:939 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 5 16 theme surface 933:939 arg1 negligible 945:954 negligible 945:954 negligible 945:954 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 6 17 theme deglycosylated 1164:1177 arg1 trimers 1179:1185 site-selective deglycosylated trimers 1149:1185 site-selective deglycosylated trimers 1149:1185 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 6 17 theme deglycosylated 1164:1177 arg1 immunogens 1198:1207 priming immunogens 1190:1207 priming immunogens 1190:1207 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 4 18 theme four-glycan-deleted 771:789 arg1 strains 804:810 over 90% breadth against four-glycan-deleted heterologous strains 746:810 over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 746:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 6 19 gly deglycosylated 1164:1177 arg1 trimers 1179:1185 site-selective deglycosylated trimers 1149:1185 site-selective deglycosylated trimers 1149:1185 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 6 19 gly deglycosylated 1164:1177 arg1 immunogens 1198:1207 priming immunogens 1190:1207 priming immunogens 1190:1207 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 2 20 theme CD4 357:359 arg1 supersite 361:369 the CD4 supersite 353:369 the CD4 supersite 353:369 Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. 28445724 5 21 theme protein 925:931 arg1 surface 933:939 the glycan-shielded protein surface 905:939 the glycan-shielded protein surface 905:939 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 4 22 theme breadth against 755:769 arg1 strains 804:810 over 90% breadth against four-glycan-deleted heterologous strains 746:810 over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 746:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 3 23 theme select 411:416 arg1 glycans 418:424 select glycans 411:424 select glycans 411:424 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 0 24 theme Impact 22:27 arg1 Quantification 0:13 Quantification 0:13 Quantification of the Impact of the HIV-1-Glycan 0:47 Quantification of the Impact of the HIV-1-Glycan Shield on Antibody Elicitation. 28445724 5 25 theme proportional 997:1008 arg1 ID50 991:994 ID50 991:994 ID50 991:994 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 5 25 theme proportional 997:1008 arg1 neutralization 975:988 Env-elicited neutralization 962:988 Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody 962:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 6 26 theme site-targeting 1238:1251 arg1 antibodies 1253:1262 site-targeting antibodies 1238:1262 site-targeting antibodies 1238:1262 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 1 27 theme quantitative 173:184 arg1 impact 186:191 its quantitative impact 169:191 its quantitative impact on antibody elicitation 169:215 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 6 28 theme antibodies 1253:1262 arg1 frequency 1225:1233 the frequency 1221:1233 the frequency of site-targeting antibodies 1221:1262 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 4 29 theme neutralization 830:843 arg1 character 845:853 tier 2 neutralization character 823:853 tier 2 neutralization character 823:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 3 30 theme rhesus 545:550 arg1 macaques 552:559 rhesus macaques 545:559 rhesus macaques 545:559 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 3 31 theme CD4 450:452 arg1 supersite 454:462 the CD4 supersite 446:462 the CD4 supersite 446:462 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 3 32 theme Env 394:396 arg1 trimers 398:404 We engineered diverse Env trimers 372:404 We engineered diverse Env trimers with select glycans 372:424 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 6 33 theme exponential 1112:1122 arg1 relationship 1124:1135 exponential relationship 1112:1135 exponential relationship 1112:1135 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 4 34 theme four-glycan-deleted 702:720 arg1 viruses 733:739 four-glycan-deleted autologous viruses 702:739 four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 702:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 5 35 theme glycan-shielded 909:923 arg1 surface 933:939 the glycan-shielded protein surface 905:939 the glycan-shielded protein surface 905:939 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 1 36 theme antibody 196:203 arg1 elicitation 205:215 antibody elicitation 196:215 antibody elicitation 196:215 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 3 37 theme We engineered 372:384 arg1 trimers 398:404 We engineered diverse Env trimers 372:404 We engineered diverse Env trimers with select glycans 372:424 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 4 38 dep strains 804:810 arg1 % 753:753 % 753:753 % 753:753 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 3 39 theme diverse 386:392 arg1 trimers 398:404 We engineered diverse Env trimers 372:404 We engineered diverse Env trimers with select glycans 372:424 We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. 28445724 5 40 with negligible 945:954 arg1 ID50 991:994 ID50 991:994 ID50 991:994 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 5 40 with negligible 945:954 arg1 neutralization 975:988 Env-elicited neutralization 962:988 Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody 962:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 2 41 theme shield 309:314 arg1 impact 288:293 the impact 284:293 the impact of the glycan shield on elicitation of antibodies against the CD4 supersite 284:369 Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. 28445724 1 42 from impact 186:191 arg1 elicitation 205:215 antibody elicitation 196:215 antibody elicitation 196:215 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 4 43 theme wild-type 614:622 arg1 viruses 624:630 wild-type viruses 614:630 wild-type viruses 614:630 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 5 44 theme accessible 1057:1066 arg1 area 1052:1055 the protein-surface area 1032:1055 the protein-surface area accessible to antibody 1032:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 4 45 theme tier 823:826 arg1 character 845:853 tier 2 neutralization character 823:853 tier 2 neutralization character 823:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 46 theme heterologous 791:802 arg1 strains 804:810 over 90% breadth against four-glycan-deleted heterologous strains 746:810 over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 746:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 1 47 theme shelter 123:129 arg1 Env 131:133 shelter Env 123:133 shelter Env from the humoral immune response 123:166 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 6 48 theme high 1096:1099 arg1 titers 1101:1106 these high titers 1090:1106 these high titers 1090:1106 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 0 49 theme Antibody 59:66 arg1 Elicitation 68:78 Antibody Elicitation 59:78 Antibody Elicitation 59:78 Quantification of the Impact of the HIV-1-Glycan Shield on Antibody Elicitation. 28445724 5 50 theme area 1052:1055 arg1 exponential 1017:1027 the exponential 1013:1027 the exponential of the protein-surface area accessible to antibody 1013:1078 To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID50) proportional to the exponential of the protein-surface area accessible to antibody. 28445724 4 51 theme CD4-supersite 643:655 arg1 neutralization 657:670 potent CD4-supersite neutralization 636:670 potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character) 636:854 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 4 51 theme CD4-supersite 643:655 arg1 titers 673:678 titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 673:853 titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character 673:853 Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). 28445724 1 52 dep HIV-1-glycan 91:102 arg1 shield 104:109 shield 104:109 shield 104:109 While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. 28445724 6 53 theme site-selective 1149:1162 arg1 trimers 1179:1185 site-selective deglycosylated trimers 1149:1185 site-selective deglycosylated trimers 1149:1185 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 6 53 theme site-selective 1149:1162 arg1 immunogens 1198:1207 priming immunogens 1190:1207 priming immunogens 1190:1207 Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. 28445724 2 54 theme antibodies 334:343 arg1 elicitation 319:329 elicitation 319:329 elicitation of antibodies against the CD4 supersite 319:369 Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. 26765751 7 0 theme EGF 1562:1564 arg1 modules 1566:1572 synthetic human NOTCH1 EGF modules 1539:1572 synthetic human NOTCH1 EGF modules 1539:1572 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 1 theme Xylα1 941:945 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 1 theme Xylα1 941:945 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 6 2 theme amino 1324:1328 arg1 residues 1335:1342 neighboring amino acid residues 1312:1342 neighboring amino acid residues 1312:1342 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 1 3 theme multicellular 263:275 arg1 organisms 277:285 multicellular organisms 263:285 multicellular organisms 263:285 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 7 4 from effects 1604:1610 arg1 adhesiveness 1637:1648 adhesiveness 1637:1648 adhesiveness 1637:1648 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 4 from effects 1604:1610 arg1 proliferation 1619:1631 proliferation 1619:1631 proliferation 1619:1631 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 5 theme human 1549:1553 arg1 modules 1566:1572 synthetic human NOTCH1 EGF modules 1539:1572 synthetic human NOTCH1 EGF modules 1539:1572 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 6 theme 3Xylα1 949:954 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 6 theme 3Xylα1 949:954 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 3 7 theme human 588:592 arg1 receptor 614:621 the human Notch transmembrane receptor 584:621 the human Notch transmembrane receptor 584:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 2 8 mod modified 398:405 arg1 signaling 385:393 Notch signaling 379:393 Notch signaling 379:393 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 2 8 mod modified 398:405 arg3 glycosylations 428:441 posttranslational glycosylations 410:441 posttranslational glycosylations of the Notch extracellular EGF-like domain 410:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 7 9 contain have 1829:1832 arg2 ability 1838:1844 the ability to interact with intrinsic Notch ligands on the surface of cancer cells 1834:1916 the ability to interact with intrinsic Notch ligands on the surface of cancer cells 1834:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 9 contain have 1829:1832 arg1 modules 1821:1827 exogenously applied synthetic EGF modules 1787:1827 exogenously applied synthetic EGF modules 1787:1827 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 6 10 theme magnetic 1196:1203 arg1 resonance 1205:1213 A nuclear magnetic resonance 1186:1213 A nuclear magnetic resonance study 1186:1219 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 6 11 theme sugar 1293:1297 arg1 moieties 1299:1306 key sugar moieties 1289:1306 key sugar moieties 1289:1306 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 0 12 theme Synthetic 0:8 arg1 Modules 27:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 1 13 theme Notch 170:174 arg1 pathway 186:192 The Notch signaling pathway 166:192 The Notch signaling pathway 166:192 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 13 theme Notch 170:174 arg1 mechanism 232:240 an evolutionarily highly conserved mechanism 197:240 an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues 197:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 13 theme Notch 170:174 arg1 critical 294:301 critical 294:301 critical 294:301 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 7 14 theme preliminary 1516:1526 arg1 test 1528:1531 A preliminary test 1514:1531 A preliminary test using synthetic human NOTCH1 EGF modules 1514:1572 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 15 theme intrinsic 1863:1871 arg1 ligands 1879:1885 intrinsic Notch ligands 1863:1885 intrinsic Notch ligands on the surface of cancer cells 1863:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 16 theme A549 1734:1737 arg1 adhesiveness 1637:1648 adhesiveness 1637:1648 adhesiveness 1637:1648 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 16 theme A549 1734:1737 arg1 proliferation 1619:1631 proliferation 1619:1631 proliferation 1619:1631 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 0 17 theme Ions 117:120 arg1 Roles 100:104 the Structural and Functional Roles 70:104 the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region 70:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 3 18 theme transmembrane 600:612 arg1 receptor 614:621 the human Notch transmembrane receptor 584:621 the human Notch transmembrane receptor 584:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 19 theme cell 1724:1727 arg1 A549 1734:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 20 theme α-L-fucopyranose-initiated 982:1007 arg1 glycan 1009:1014 α-L-fucopyranose-initiated glycan 982:1014 α-L-fucopyranose-initiated glycan 982:1014 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 6 21 theme effects 1273:1279 arg1 existence 1234:1242 the existence 1230:1242 the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues 1230:1342 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 0 22 theme O-Glycans 126:134 arg1 Roles 100:104 the Structural and Functional Roles 70:104 the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region 70:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 5 23 theme glycosylated 1114:1125 arg1 EGF12 1127:1131 the glycosylated EGF12 1110:1131 the glycosylated EGF12 1110:1131 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 3 24 theme universal 641:649 arg1 method 651:656 a universal method 639:656 a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites 639:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 25 theme adenocarcinoma 1698:1711 arg1 A549 1734:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 26 theme 3Glcβ1 958:963 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 26 theme 3Glcβ1 958:963 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 6 27 theme nuclear 1254:1260 arg1 effects 1273:1279 strong nuclear Overhauser effects 1247:1279 strong nuclear Overhauser effects 1247:1279 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 5 28 theme proper 1092:1097 arg1 folding 1099:1105 the proper folding 1088:1105 the proper folding of the glycosylated EGF12 1088:1131 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 7 29 theme line 1678:1681 arg1 MCF-7 1683:1687 human breast cancer cell line MCF-7 1653:1687 human breast cancer cell line MCF-7 1653:1687 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 30 theme structural 503:512 arg1 roles 529:533 the structural and functional roles 499:533 the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor 499:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 6 31 theme antiparallel 1444:1455 arg1 structure 1465:1473 the antiparallel β-sheet structure 1440:1473 the antiparallel β-sheet structure in the ligand-binding region of EGF12 1440:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 0 32 theme NOTCH1 16:21 arg1 Modules 27:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 7 33 theme synthetic 1807:1815 arg1 modules 1821:1827 exogenously applied synthetic EGF modules 1787:1827 exogenously applied synthetic EGF modules 1787:1827 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 34 theme cancer 1666:1671 arg1 MCF-7 1683:1687 human breast cancer cell line MCF-7 1653:1687 human breast cancer cell line MCF-7 1653:1687 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 0 35 theme Structural 74:83 arg1 Roles 100:104 the Structural and Functional Roles 70:104 the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region 70:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 1 36 theme most 365:368 arg1 tissues 370:376 most tissues 365:376 most tissues 365:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 7 37 theme human 1653:1657 arg1 MCF-7 1683:1687 human breast cancer cell line MCF-7 1653:1687 human breast cancer cell line MCF-7 1653:1687 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 0 38 theme Functional 89:98 arg1 Roles 100:104 the Structural and Functional Roles 70:104 the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region 70:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 4 39 theme rapid 830:834 arg1 synthesis 857:865 the rapid and highly efficient synthesis 826:865 the rapid and highly efficient synthesis of NOTCH1 EGF12 826:881 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 6 40 from stabilization 1423:1435 arg1 region 1497:1502 the ligand-binding region 1478:1502 the ligand-binding region of EGF12 1478:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 7 41 theme first 1762:1766 arg1 time 1768:1771 the first time 1758:1771 the first time 1758:1771 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 6 42 theme EGF12 1507:1511 arg1 region 1497:1502 the ligand-binding region 1478:1502 the ligand-binding region of EGF12 1478:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 2 43 theme domain 479:484 arg1 glycosylations 428:441 posttranslational glycosylations 410:441 posttranslational glycosylations of the Notch extracellular EGF-like domain 410:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 2 44 theme extracellular 456:468 arg1 domain 479:484 the Notch extracellular EGF-like domain 446:484 the Notch extracellular EGF-like domain 446:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 4 45 dep having 896:901 arg1 → 1026:1026 → 1026:1026 → 1026:1026 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 3 46 theme modules 693:699 arg1 construction 666:677 the construction 662:677 the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites 662:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 4 47 theme NOTCH1 870:875 arg1 EGF12 877:881 NOTCH1 EGF12 870:881 NOTCH1 EGF12 870:881 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 5 48 theme mM 1176:1177 arg1 CaCl2 1179:1183 5 mM CaCl2 1174:1183 5 mM CaCl2 1174:1183 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 3 49 theme NOTCH1 682:687 arg1 modules 693:699 NOTCH1 EGF modules 682:699 NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites 682:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 6 50 theme structure 1465:1473 arg1 stabilization 1423:1435 the intramolecular stabilization 1404:1435 the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12 1404:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 7 51 theme significant 1581:1591 arg1 effects 1604:1610 significant inhibitory effects 1581:1610 significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549 1581:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 52 theme functional 518:527 arg1 roles 529:533 the structural and functional roles 499:533 the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor 499:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 1 53 theme conserved 222:230 arg1 pathway 186:192 The Notch signaling pathway 166:192 The Notch signaling pathway 166:192 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 53 theme conserved 222:230 arg1 mechanism 232:240 an evolutionarily highly conserved mechanism 197:240 an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues 197:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 53 theme conserved 222:230 arg1 critical 294:301 critical 294:301 critical 294:301 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 7 54 theme cancer 1905:1910 arg1 cells 1912:1916 cancer cells 1905:1916 cancer cells 1905:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 55 theme multiple 560:567 arg1 domains 573:579 multiple EGF domains 560:579 multiple EGF domains in the human Notch transmembrane receptor 560:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 56 theme NOTCH1 1555:1560 arg1 modules 1566:1572 synthetic human NOTCH1 EGF modules 1539:1572 synthetic human NOTCH1 EGF modules 1539:1572 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 57 from ligands 1879:1885 arg1 surface 1894:1900 the surface 1890:1900 the surface of cancer cells 1890:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 58 theme glycosylation 752:764 arg1 sites 766:770 the designated glycosylation sites 737:770 the designated glycosylation sites 737:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 1 59 from development 334:344 arg1 tissues 370:376 most tissues 365:376 most tissues 365:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 2 60 theme Notch 379:383 arg1 signaling 385:393 Notch signaling 379:393 Notch signaling 379:393 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 7 61 theme synthetic 1539:1547 arg1 modules 1566:1572 synthetic human NOTCH1 EGF modules 1539:1572 synthetic human NOTCH1 EGF modules 1539:1572 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 6 62 theme neighboring 1312:1322 arg1 residues 1335:1342 neighboring amino acid residues 1312:1342 neighboring amino acid residues 1312:1342 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 4 63 theme → 947:947 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 63 theme → 947:947 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 7 64 theme Notch 1873:1877 arg1 ligands 1879:1885 intrinsic Notch ligands 1863:1885 intrinsic Notch ligands on the surface of cancer cells 1863:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 6 65 theme resonance 1205:1213 arg1 study 1215:1219 A nuclear magnetic resonance study 1186:1219 A nuclear magnetic resonance study 1186:1219 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 0 66 theme Molecular 45:53 arg1 Mechanisms 55:64 Molecular Mechanisms 45:64 Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region 45:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 6 67 theme nuclear 1188:1194 arg1 resonance 1205:1213 A nuclear magnetic resonance 1186:1213 A nuclear magnetic resonance study 1186:1219 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 6 68 theme key 1289:1291 arg1 moieties 1299:1306 key sugar moieties 1289:1306 key sugar moieties 1289:1306 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 1 69 theme signaling 176:184 arg1 pathway 186:192 The Notch signaling pathway 166:192 The Notch signaling pathway 166:192 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 69 theme signaling 176:184 arg1 mechanism 232:240 an evolutionarily highly conserved mechanism 197:240 an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues 197:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 1 69 theme signaling 176:184 arg1 critical 294:301 critical 294:301 critical 294:301 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 7 70 theme line 1729:1732 arg1 A549 1734:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 71 theme β-D-glucopyranose-initiated 905:931 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 71 theme β-D-glucopyranose-initiated 905:931 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 72 dep → 1026:1026 arg1 4GlcNAcβ1 1037:1045 → 4GlcNAcβ1 → 3Fucα1 → 1035:1056 Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 → 1017:1056 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 3 73 theme Notch 594:598 arg1 receptor 614:621 the human Notch transmembrane receptor 584:621 the human Notch transmembrane receptor 584:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 74 theme epithelial 1713:1722 arg1 A549 1734:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 75 theme glycoforms 546:555 arg1 roles 529:533 the structural and functional roles 499:533 the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor 499:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 6 76 from region 1497:1502 arg1 stabilization 1423:1435 the intramolecular stabilization 1404:1435 the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12 1404:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 6 77 theme acid 1330:1333 arg1 residues 1335:1342 neighboring amino acid residues 1312:1342 neighboring amino acid residues 1312:1342 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 7 78 theme lung 1693:1696 arg1 A549 1734:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 lung adenocarcinoma epithelial cell line A549 1693:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 6 79 theme Overhauser 1262:1271 arg1 effects 1273:1279 strong nuclear Overhauser effects 1247:1279 strong nuclear Overhauser effects 1247:1279 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 0 80 theme Ligand-Binding 143:156 arg1 Region 158:163 the Ligand-Binding Region 139:163 the Ligand-Binding Region 139:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 5 81 theme EGF12 1127:1131 arg1 folding 1099:1105 the proper folding 1088:1105 the proper folding of the glycosylated EGF12 1088:1131 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 7 82 theme MCF-7 1683:1687 arg1 adhesiveness 1637:1648 adhesiveness 1637:1648 adhesiveness 1637:1648 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 82 theme MCF-7 1683:1687 arg1 proliferation 1619:1631 proliferation 1619:1631 proliferation 1619:1631 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 4 83 theme → 956:956 arg1 → 965:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 Xylα1 → 3Xylα1 → 3Glcβ1 → 941:965 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 4 83 theme → 956:956 arg1 glycan 933:938 a β-D-glucopyranose-initiated glycan 903:938 a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) 903:966 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 6 84 theme strong 1247:1252 arg1 effects 1273:1279 strong nuclear Overhauser effects 1247:1279 strong nuclear Overhauser effects 1247:1279 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 6 85 from structure 1465:1473 arg1 region 1497:1502 the ligand-binding region 1478:1502 the ligand-binding region of EGF12 1478:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 7 86 theme cell 1673:1676 arg1 MCF-7 1683:1687 human breast cancer cell line MCF-7 1653:1687 human breast cancer cell line MCF-7 1653:1687 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 0 87 from Roles 100:104 arg1 Region 158:163 the Ligand-Binding Region 139:163 the Ligand-Binding Region 139:163 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 6 88 theme β-sheet 1457:1463 arg1 structure 1465:1473 the antiparallel β-sheet structure 1440:1473 the antiparallel β-sheet structure in the ligand-binding region of EGF12 1440:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 5 89 theme folding 1099:1105 arg1 efficiency 1074:1083 The efficiency 1070:1083 The efficiency of the proper folding of the glycosylated EGF12 1070:1131 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 7 90 theme breast 1659:1664 arg1 MCF-7 1683:1687 human breast cancer cell line MCF-7 1653:1687 human breast cancer cell line MCF-7 1653:1687 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 3 91 from domains 573:579 arg1 receptor 614:621 the human Notch transmembrane receptor 584:621 the human Notch transmembrane receptor 584:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 3 91 from domains 573:579 arg1 roles 529:533 the structural and functional roles 499:533 the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor 499:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 6 92 theme intramolecular 1408:1421 arg1 stabilization 1423:1435 the intramolecular stabilization 1404:1435 the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12 1404:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 7 93 theme EGF 1817:1819 arg1 modules 1821:1827 exogenously applied synthetic EGF modules 1787:1827 exogenously applied synthetic EGF modules 1787:1827 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 5 94 theme 5 1174:1174 arg1 mM 1176:1177 mM 1176:1177 mM 1176:1177 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 7 95 theme applied 1799:1805 arg1 modules 1821:1827 exogenously applied synthetic EGF modules 1787:1827 exogenously applied synthetic EGF modules 1787:1827 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 0 96 theme Calcium 109:115 arg1 Ions 117:120 Calcium Ions 109:120 Calcium Ions 109:120 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 2 97 gly glycosylations 428:441 arg1 domain 479:484 the Notch extracellular EGF-like domain 446:484 the Notch extracellular EGF-like domain 446:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 2 98 theme posttranslational 410:426 arg1 glycosylations 428:441 posttranslational glycosylations 410:441 posttranslational glycosylations of the Notch extracellular EGF-like domain 410:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 0 99 theme Human 10:14 arg1 Modules 27:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 0 100 theme EGF 23:25 arg1 Modules 27:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules 0:33 Synthetic Human NOTCH1 EGF Modules Unraveled Molecular Mechanisms for the Structural and Functional Roles of Calcium Ions and O-Glycans in the Ligand-Binding Region. 26765751 2 101 theme EGF-like 470:477 arg1 domain 479:484 the Notch extracellular EGF-like domain 446:484 the Notch extracellular EGF-like domain 446:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 3 102 theme EGF 689:691 arg1 modules 693:699 NOTCH1 EGF modules 682:699 NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites 682:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 4 103 theme strategy 797:804 arg1 versatility 777:787 The versatility 773:787 The versatility of this strategy 773:804 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 2 104 theme Notch 450:454 arg1 domain 479:484 the Notch extracellular EGF-like domain 446:484 the Notch extracellular EGF-like domain 446:484 Notch signaling is modified by posttranslational glycosylations of the Notch extracellular EGF-like domain. 26765751 6 105 theme ligand-binding 1482:1495 arg1 region 1497:1502 the ligand-binding region 1478:1502 the ligand-binding region of EGF12 1478:1511 A nuclear magnetic resonance study revealed the existence of strong nuclear Overhauser effects between key sugar moieties and neighboring amino acid residues, indicating that both O-glycans contribute independently to the intramolecular stabilization of the antiparallel β-sheet structure in the ligand-binding region of EGF12. 26765751 1 106 theme cell-fate 307:315 arg1 decisions 317:325 cell-fate decisions 307:325 cell-fate decisions during development and homeostasis in most tissues 307:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 3 107 gly glycosylation 752:764 arg2 sites 766:770 the designated glycosylation sites 737:770 the designated glycosylation sites 737:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 1 108 from homeostasis 350:360 arg1 tissues 370:376 most tissues 365:376 most tissues 365:376 The Notch signaling pathway is an evolutionarily highly conserved mechanism that operates across multicellular organisms and is critical for cell-fate decisions during development and homeostasis in most tissues. 26765751 5 109 theme CaCl2 1179:1183 arg1 presence 1162:1169 the presence 1158:1169 the presence of 5 mM CaCl2 1158:1183 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 3 110 theme desired 716:722 arg1 O-glycans 724:732 the desired O-glycans 712:732 the desired O-glycans 712:732 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 4 111 theme EGF12 877:881 arg1 synthesis 857:865 the rapid and highly efficient synthesis 826:865 the rapid and highly efficient synthesis of NOTCH1 EGF12 826:881 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 3 112 theme various 538:544 arg1 glycoforms 546:555 various glycoforms 538:555 various glycoforms 538:555 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 113 theme inhibitory 1593:1602 arg1 effects 1604:1610 significant inhibitory effects 1581:1610 significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549 1581:1737 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 7 114 theme cells 1912:1916 arg1 surface 1894:1900 the surface 1890:1900 the surface of cancer cells 1890:1916 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 26765751 5 115 gly glycosylated 1114:1125 arg1 EGF12 1127:1131 the glycosylated EGF12 1110:1131 the glycosylated EGF12 1110:1131 The efficiency of the proper folding of the glycosylated EGF12 was markedly enhanced in the presence of 5 mM CaCl2. 26765751 3 116 theme designated 741:750 arg1 sites 766:770 the designated glycosylation sites 737:770 the designated glycosylation sites 737:770 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 4 117 theme efficient 847:855 arg1 synthesis 857:865 the rapid and highly efficient synthesis 826:865 the rapid and highly efficient synthesis of NOTCH1 EGF12 826:881 The versatility of this strategy was demonstrated by the rapid and highly efficient synthesis of NOTCH1 EGF12 concurrently having a β-D-glucopyranose-initiated glycan (Xylα1 → 3Xylα1 → 3Glcβ1 →) at Ser458 and α-L-fucopyranose-initiated glycan (Neu5Acα2 → 3Galβ1 → 4GlcNAcβ1 → 3Fucα1 →) at Thr466. 26765751 3 118 theme EGF 569:571 arg1 domains 573:579 multiple EGF domains 560:579 multiple EGF domains in the human Notch transmembrane receptor 560:621 To evaluate the structural and functional roles of various glycoforms at multiple EGF domains in the human Notch transmembrane receptor, we established a universal method for the construction of NOTCH1 EGF modules displaying the desired O-glycans at the designated glycosylation sites. 26765751 7 119 dep proliferation 1619:1631 arg1 the 1615:1617 the 1615:1617 the 1615:1617 A preliminary test using synthetic human NOTCH1 EGF modules showed significant inhibitory effects on the proliferation and adhesiveness of human breast cancer cell line MCF-7 and lung adenocarcinoma epithelial cell line A549, demonstrating for the first time evidence that exogenously applied synthetic EGF modules have the ability to interact with intrinsic Notch ligands on the surface of cancer cells. 25265424 7 0 theme enzymatic 1265:1273 arg1 activity 1275:1282 enzymatic activity 1265:1282 enzymatic activity 1265:1282 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 6 1 theme trace 915:919 arg1 amounts 921:927 only trace amounts 910:927 only trace amounts in haptoglobin 910:942 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 5 2 theme detached 693:700 arg1 glycans 702:708 permethylated detached glycans 679:708 permethylated detached glycans 679:708 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 2 3 theme serologic 282:290 arg1 monitoring 292:301 noninvasive serologic monitoring 270:301 noninvasive serologic monitoring 270:301 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 0 4 from specificity 17:27 arg1 glycoproteins 63:75 liver-secreted glycoproteins 48:75 liver-secreted glycoproteins 48:75 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 5 5 theme MSn 663:665 arg1 analysis 667:674 MSn analysis 663:674 MSn analysis of permethylated detached glycans 663:708 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 6 6 theme complement 981:990 arg1 H 999:999 complement factor H 981:999 complement factor H 981:999 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 5 7 theme LeY 735:737 arg1 glycoforms 739:748 LeY glycoforms 735:748 LeY glycoforms 735:748 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 8 8 theme functional 1505:1514 arg1 consequences 1516:1527 functional consequences 1505:1527 functional consequences for specific subsets of proteins or their domains 1505:1577 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 4 9 gly fucosylation 561:572 arg1 some 577:580 some 577:580 some 577:580 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 4 9 gly fucosylation 561:572 arg1 proteins 589:596 the proteins 585:596 the proteins 585:596 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 5 10 theme H 863:863 arg1 epitopes 865:872 H epitopes 863:872 H epitopes 863:872 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 8 11 theme proteins 1553:1560 arg1 subsets 1542:1548 specific subsets 1533:1548 specific subsets of proteins or their domains 1533:1577 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 7 12 theme same 1195:1198 arg1 source 1208:1213 the same patient source 1191:1213 the same patient source 1191:1213 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 3 13 theme case 317:320 arg1 study 322:326 a case study 315:326 a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient 315:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 4 14 theme exoglycosidase-assisted 491:513 arg1 analysis 524:531 Our exoglycosidase-assisted LC-MS/MS analysis 487:531 Our exoglycosidase-assisted LC-MS/MS analysis 487:531 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 8 15 theme proteins 1380:1387 arg1 selection 1357:1365 selection 1357:1365 selection of candidate proteins for disease monitoring 1357:1410 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 0 16 from Protein 0:6 arg1 glycoproteins 63:75 liver-secreted glycoproteins 48:75 liver-secreted glycoproteins 48:75 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 7 17 attach isolated 1177:1184 arg2 proteins 1168:1175 proteins 1168:1175 proteins isolated from the same patient source 1168:1213 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 7 17 attach isolated 1177:1184 arg1 source 1208:1213 the same patient source 1191:1213 the same patient source 1191:1213 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 8 18 theme disease 1393:1399 arg1 monitoring 1401:1410 disease monitoring 1393:1410 disease monitoring 1393:1410 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 1 19 theme liver 86:90 arg1 problem 122:128 a serious health problem worldwide 105:138 a serious health problem worldwide 105:138 Chronic liver diseases are a serious health problem worldwide. 25265424 1 19 theme liver 86:90 arg1 diseases 92:99 Chronic liver diseases 78:99 Chronic liver diseases 78:99 Chronic liver diseases are a serious health problem worldwide. 25265424 6 20 theme % 1091:1091 arg1 intensity 1093:1101 15-20% intensity 1086:1101 15-20% intensity 1086:1101 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 4 21 theme some 577:580 arg1 fucosylation 561:572 fucosylation 561:572 fucosylation of some of the proteins 561:596 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 4 22 theme fucosylation 561:572 arg1 degree 551:556 the high degree 542:556 the high degree of fucosylation of some of the proteins 542:596 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 5 23 contain carry 847:851 arg2 epitopes 865:872 H epitopes 863:872 H epitopes 863:872 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 23 contain carry 847:851 arg2 Lewis 853:857 Lewis 853:857 Lewis 853:857 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 23 contain carry 847:851 arg1 proteins 838:845 all three proteins 828:845 all three proteins 828:845 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 0 24 theme site 12:15 arg1 specificity 17:27 site specificity 12:27 site specificity 12:27 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 3 25 gly glycoforms 356:365 arg1 haptoglobin 394:404 haptoglobin 394:404 haptoglobin 394:404 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 3 25 gly glycoforms 356:365 arg1 hemopexin 445:453 hemopexin 445:453 hemopexin isolated from the same patient 445:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 3 25 gly glycoforms 356:365 arg1 proteins 375:382 four proteins 370:382 four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient 370:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 3 25 gly glycoforms 356:365 arg1 kininogen-1 428:438 kininogen-1 428:438 kininogen-1 428:438 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 3 25 gly glycoforms 356:365 arg1 H 425:425 complement factor H 407:425 complement factor H 407:425 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 1 26 theme serious 107:113 arg1 problem 122:128 a serious health problem worldwide 105:138 a serious health problem worldwide 105:138 Chronic liver diseases are a serious health problem worldwide. 25265424 1 26 theme serious 107:113 arg1 diseases 92:99 Chronic liver diseases 78:99 Chronic liver diseases 78:99 Chronic liver diseases are a serious health problem worldwide. 25265424 6 27 theme select 1057:1062 arg1 glycopeptides 1064:1076 select glycopeptides 1057:1076 select glycopeptides 1057:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 3 28 theme factor 418:423 arg1 H 425:425 complement factor H 407:425 complement factor H 407:425 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 7 29 from differences 1127:1137 arg1 fucosylation 1142:1153 fucosylation 1142:1153 fucosylation 1142:1153 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 2 30 theme alterations 179:189 arg1 alterations 179:189 the frequently reported glycan alterations 148:189 the frequently reported glycan alterations in liver disease 148:206 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 2 30 theme alterations 179:189 arg1 One 141:143 One 141:143 One 141:143 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 8 31 theme structural 1460:1469 arg1 determinants 1471:1482 structural determinants 1460:1482 structural determinants 1460:1482 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 5 32 theme complement 807:816 arg1 H 825:825 complement factor H 807:825 complement factor H 807:825 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 6 33 theme glycans 1046:1052 arg1 fucosylation 1013:1024 core fucosylation 1008:1024 core fucosylation of the bi-antennary glycans on select glycopeptides 1008:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 8 34 contain have 1455:1458 arg2 determinants 1471:1482 structural determinants 1460:1482 structural determinants 1460:1482 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 8 34 contain have 1455:1458 arg1 glycoforms 1444:1453 site-specific glycoforms 1430:1453 site-specific glycoforms 1430:1453 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 2 35 theme reported 163:170 arg1 alterations 179:189 the frequently reported glycan alterations 148:189 the frequently reported glycan alterations in liver disease 148:206 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 8 36 theme domains 1571:1577 arg1 subsets 1542:1548 specific subsets 1533:1548 specific subsets of proteins or their domains 1533:1577 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 3 37 theme same 473:476 arg1 patient 478:484 the same patient 469:484 the same patient 469:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 7 38 gly microheterogeneity 1297:1314 arg1 glycoforms 1319:1328 glycoforms 1319:1328 glycoforms 1319:1328 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 6 39 from amounts 921:927 arg1 detectable 896:905 detectable 896:905 detectable 896:905 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 6 39 from amounts 921:927 arg1 haptoglobin 932:942 haptoglobin 932:942 haptoglobin 932:942 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 2 40 theme liver 194:198 arg1 disease 200:206 liver disease 194:206 liver disease 194:206 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 5 41 theme factor 818:823 arg1 H 825:825 complement factor H 807:825 complement factor H 807:825 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 2 42 from alterations 179:189 arg1 disease 200:206 liver disease 194:206 liver disease 194:206 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 7 43 theme activity 1275:1282 arg1 up-regulation 1248:1260 up-regulation 1248:1260 up-regulation of enzymatic activity 1248:1282 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 6 44 from detectable 896:905 arg1 amounts 921:927 only trace amounts 910:927 only trace amounts in haptoglobin 910:942 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 5 45 theme permethylated 679:691 arg1 glycans 702:708 permethylated detached glycans 679:708 permethylated detached glycans 679:708 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 3 46 attach isolated 455:462 arg2 hemopexin 445:453 hemopexin 445:453 hemopexin isolated from the same patient 445:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 3 46 attach isolated 455:462 arg1 patient 478:484 the same patient 469:484 the same patient 469:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 7 47 theme protein-specific 1110:1125 arg1 differences 1127:1137 These protein-specific differences 1104:1137 These protein-specific differences 1104:1137 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 2 48 theme noninvasive 270:280 arg1 monitoring 292:301 noninvasive serologic monitoring 270:301 noninvasive serologic monitoring 270:301 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 6 49 theme factor 992:997 arg1 H 999:999 complement factor H 981:999 complement factor H 981:999 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 5 50 theme glycoforms 739:748 arg1 presence 723:730 the presence 719:730 the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H 719:825 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 8 51 theme specific 1533:1540 arg1 subsets 1542:1548 specific subsets 1533:1548 specific subsets of proteins or their domains 1533:1577 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 5 52 theme glycans 702:708 arg1 analysis 667:674 MSn analysis 663:674 MSn analysis of permethylated detached glycans 663:708 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 1 53 theme health 115:120 arg1 problem 122:128 a serious health problem worldwide 105:138 a serious health problem worldwide 105:138 Chronic liver diseases are a serious health problem worldwide. 25265424 1 53 theme health 115:120 arg1 diseases 92:99 Chronic liver diseases 78:99 Chronic liver diseases 78:99 Chronic liver diseases are a serious health problem worldwide. 25265424 1 54 theme worldwide 130:138 arg1 problem 122:128 a serious health problem worldwide 105:138 a serious health problem worldwide 105:138 Chronic liver diseases are a serious health problem worldwide. 25265424 1 54 theme worldwide 130:138 arg1 diseases 92:99 Chronic liver diseases 78:99 Chronic liver diseases 78:99 Chronic liver diseases are a serious health problem worldwide. 25265424 6 55 theme bi-antennary 1033:1044 arg1 glycans 1046:1052 the bi-antennary glycans 1029:1052 the bi-antennary glycans 1029:1052 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 8 56 contain has 1336:1338 arg1 This 1331:1334 This 1331:1334 This 1331:1334 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 8 56 contain has 1336:1338 arg2 implications 1340:1351 implications 1340:1351 implications for selection of candidate proteins for disease monitoring 1340:1410 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 5 57 gly glycoforms 739:748 arg1 LeY 735:737 LeY glycoforms 735:748 LeY glycoforms 735:748 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 4 58 theme LC-MS/MS 515:522 arg1 analysis 524:531 Our exoglycosidase-assisted LC-MS/MS analysis 487:531 Our exoglycosidase-assisted LC-MS/MS analysis 487:531 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 7 59 located observed 1156:1163 arg2 differences 1127:1137 These protein-specific differences 1104:1137 These protein-specific differences 1104:1137 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 7 59 located observed 1156:1163 arg1 proteins 1168:1175 proteins 1168:1175 proteins isolated from the same patient source 1168:1213 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 7 60 theme other 1237:1241 arg1 factors 1229:1235 factors 1229:1235 factors other than up-regulation of enzymatic activity 1229:1282 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 6 61 theme core 1008:1011 arg1 fucosylation 1013:1024 core fucosylation 1008:1024 core fucosylation of the bi-antennary glycans on select glycopeptides 1008:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 6 62 from fucosylation 1013:1024 arg1 glycopeptides 1064:1076 select glycopeptides 1057:1076 select glycopeptides 1057:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 8 63 theme candidate 1370:1378 arg1 proteins 1380:1387 candidate proteins 1370:1387 candidate proteins 1370:1387 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 0 64 gly glycoproteins 63:75 arg1 glycoproteins 63:75 liver-secreted glycoproteins 48:75 liver-secreted glycoproteins 48:75 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 1 65 theme Chronic 78:84 arg1 problem 122:128 a serious health problem worldwide 105:138 a serious health problem worldwide 105:138 Chronic liver diseases are a serious health problem worldwide. 25265424 1 65 theme Chronic 78:84 arg1 diseases 92:99 Chronic liver diseases 78:99 Chronic liver diseases 78:99 Chronic liver diseases are a serious health problem worldwide. 25265424 3 66 theme specific 347:354 arg1 glycoforms 356:365 site specific glycoforms 342:365 site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient 342:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 7 67 theme patient 1200:1206 arg1 source 1208:1213 the same patient source 1191:1213 the same patient source 1191:1213 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 6 68 theme 15-20 1086:1090 arg1 % 1091:1091 % 1091:1091 % 1091:1091 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 3 69 theme proteins 375:382 arg1 glycoforms 356:365 site specific glycoforms 342:365 site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient 342:484 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 6 70 gly glycopeptides 1064:1076 arg2 glycopeptides 1064:1076 select glycopeptides 1057:1076 select glycopeptides 1057:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 8 71 theme site-specific 1430:1442 arg1 glycoforms 1444:1453 site-specific glycoforms 1430:1453 site-specific glycoforms 1430:1453 This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains. 25265424 5 72 attach presence 723:730 arg1 haptoglobin 753:763 haptoglobin 753:763 haptoglobin 753:763 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 72 attach presence 723:730 arg2 glycoforms 739:748 LeY glycoforms 735:748 LeY glycoforms 735:748 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 0 73 theme fucosylation 32:43 arg1 Protein 0:6 Protein 0:6 Protein 0:6 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 0 73 theme fucosylation 32:43 arg1 specificity 17:27 site specificity 12:27 site specificity 12:27 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 6 74 theme Core 875:878 arg1 fucosylation 880:891 Core fucosylation 875:891 Core fucosylation 875:891 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 4 75 theme high 546:549 arg1 degree 551:556 the high degree 542:556 the high degree of fucosylation of some of the proteins 542:596 Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. 25265424 2 76 theme glycan 172:177 arg1 alterations 179:189 the frequently reported glycan alterations 148:189 the frequently reported glycan alterations in liver disease 148:206 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 0 77 theme liver-secreted 48:61 arg1 glycoproteins 63:75 liver-secreted glycoproteins 48:75 liver-secreted glycoproteins 48:75 Protein and site specificity of fucosylation in liver-secreted glycoproteins. 25265424 7 78 theme glycoforms 1319:1328 arg1 microheterogeneity 1297:1314 the microheterogeneity 1293:1314 the microheterogeneity of glycoforms 1293:1328 These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. 25265424 3 79 theme complement 407:416 arg1 H 425:425 complement factor H 407:425 complement factor H 407:425 We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. 25265424 2 80 theme aberrant 211:218 arg1 fucosylation 220:231 aberrant fucosylation 211:231 aberrant fucosylation 211:231 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 2 80 theme aberrant 211:218 arg1 marker 259:264 a marker 257:264 a marker for noninvasive serologic monitoring 257:301 One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. 25265424 6 81 from confidence 953:962 arg1 H 999:999 complement factor H 981:999 complement factor H 981:999 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 6 81 from confidence 953:962 arg1 hemopexin 967:975 hemopexin 967:975 hemopexin 967:975 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 5 82 located detected 782:789 arg2 haptoglobin 753:763 haptoglobin 753:763 haptoglobin 753:763 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 82 located detected 782:789 arg1 H 825:825 complement factor H 807:825 complement factor H 807:825 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 82 located detected 782:789 arg1 hemopexin 794:802 hemopexin 794:802 hemopexin 794:802 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 5 83 from presence 723:730 arg1 haptoglobin 753:763 haptoglobin 753:763 haptoglobin 753:763 MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. 25265424 6 84 with detectable 896:905 arg1 confidence 953:962 confidence 953:962 confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity 953:1101 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 6 85 gly fucosylation 1013:1024 arg1 glycans 1046:1052 the bi-antennary glycans 1029:1052 the bi-antennary glycans 1029:1052 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 25265424 6 85 gly fucosylation 1013:1024 arg1 glycopeptides 1064:1076 select glycopeptides 1057:1076 select glycopeptides 1057:1076 Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. 27649061 6 0 theme detection 906:914 arg1 specificity 891:901 specificity 891:901 specificity of detection 891:914 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 7 1 theme IgG4 1223:1226 arg1 glycoforms 1156:1165 26 glycoforms 1153:1165 26 glycoforms of IgG1 1153:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 1 theme IgG4 1223:1226 arg1 glycoforms 1209:1218 19 glycoforms 1206:1218 19 glycoforms of IgG4 1206:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 1 theme IgG4 1223:1226 arg1 glycoforms 1179:1188 22 glycoforms 1176:1188 22 glycoforms of IgG 2/3 1176:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 2 theme IgG 1148:1150 arg1 glycoforms 1134:1143 previously reported glycoforms 1114:1143 previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1114:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 5 3 dep 60 798:799 arg1 to 795:796 to 795:796 to 795:796 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 4 4 theme independent 598:608 arg1 DIA 623:625 DIA 623:625 DIA 623:625 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 4 4 theme independent 598:608 arg1 acquisition 610:620 data independent acquisition 593:620 data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions 593:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 7 5 theme glycoforms 1134:1143 arg1 majority 1102:1109 the majority 1098:1109 the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1098:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 9 6 theme IgG 1674:1676 arg1 glycoforms 1678:1687 IgG glycoforms 1674:1687 IgG glycoforms 1674:1687 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 4 7 theme minimal 655:661 arg1 conditions 678:687 "minimal" fragmentation conditions 654:687 "minimal" fragmentation conditions 654:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 2 8 theme microheterogeneity 278:295 arg1 determination 244:256 the determination 240:256 the determination of the IgG N-glycan microheterogeneity 240:295 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 6 9 theme high 866:869 arg1 resolution 871:880 high resolution 866:880 high resolution 866:880 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 4 10 theme fragmentation 664:676 arg1 conditions 678:687 "minimal" fragmentation conditions 654:687 "minimal" fragmentation conditions 654:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 7 11 theme majority 1102:1109 arg1 quantification 1080:1093 label-free quantification 1069:1093 label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1069:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 4 12 theme Y-ions 631:636 arg1 DIA 623:625 DIA 623:625 DIA 623:625 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 4 12 theme Y-ions 631:636 arg1 acquisition 610:620 data independent acquisition 593:620 data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions 593:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 7 13 dep IgG 1148:1150 arg1 glycoforms 1156:1165 26 glycoforms 1153:1165 26 glycoforms of IgG1 1153:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 13 dep IgG 1148:1150 arg1 glycoforms 1209:1218 19 glycoforms 1206:1218 19 glycoforms of IgG4 1206:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 13 dep IgG 1148:1150 arg1 glycoforms 1179:1188 22 glycoforms 1176:1188 22 glycoforms of IgG 2/3 1176:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 5 14 theme induced 713:719 arg1 conditions 740:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 8 15 from quantification 1414:1427 arg1 samples 1464:1470 pooled plasma samples 1450:1470 pooled plasma samples of patients with liver cirrhosis 1450:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 8 16 theme glycoforms 1436:1445 arg1 quantification 1414:1427 The SWATH data independent quantification 1387:1427 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis 1387:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 7 17 theme fucosylated 1363:1373 arg1 glycoforms 1375:1384 doubly fucosylated glycoforms 1356:1384 doubly fucosylated glycoforms 1356:1384 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 18 gly glycoforms 1134:1143 arg1 IgG 1148:1150 IgG 1148:1150 IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1148:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 19 theme label-free 1069:1078 arg1 quantification 1080:1093 label-free quantification 1069:1093 label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1069:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 20 theme IgG1 1170:1173 arg1 glycoforms 1156:1165 26 glycoforms 1153:1165 26 glycoforms of IgG1 1153:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 20 theme IgG1 1170:1173 arg1 glycoforms 1209:1218 19 glycoforms 1206:1218 19 glycoforms of IgG4 1206:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 20 theme IgG1 1170:1173 arg1 glycoforms 1179:1188 22 glycoforms 1176:1188 22 glycoforms of IgG 2/3 1176:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 8 21 theme pooled 1450:1455 arg1 samples 1464:1470 pooled plasma samples 1450:1470 pooled plasma samples of patients with liver cirrhosis 1450:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 7 22 theme IgG1 1340:1343 arg1 glycoforms 1326:1335 previously unreported glycoforms 1304:1335 previously unreported glycoforms of IgG1 1304:1343 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 5 23 theme adjusted 694:701 arg1 conditions 740:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 2 24 theme IgG 265:267 arg1 microheterogeneity 278:295 the IgG N-glycan microheterogeneity 261:295 the IgG N-glycan microheterogeneity 261:295 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 8 25 theme patients 1475:1482 arg1 samples 1464:1470 pooled plasma samples 1450:1470 pooled plasma samples of patients with liver cirrhosis 1450:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 7 26 theme glycoforms 1326:1335 arg1 traces 1294:1299 traces 1294:1299 traces 1294:1299 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 26 theme glycoforms 1326:1335 arg1 glycoforms 1375:1384 doubly fucosylated glycoforms 1356:1384 doubly fucosylated glycoforms 1356:1384 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 6 27 theme signal-to-noise 1006:1020 arg1 ratios 1022:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 0 28 from Analysis 17:24 arg1 Samples 47:53 Samples 47:53 Samples of Unfractionated Human Plasma 47:84 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 0 29 theme Human 73:77 arg1 Plasma 79:84 Unfractionated Human Plasma 58:84 Unfractionated Human Plasma 58:84 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 6 30 theme overall 978:984 arg1 intensity 986:994 overall intensity 978:994 overall intensity 978:994 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 7 31 theme IgG 1193:1195 arg1 glycoforms 1156:1165 26 glycoforms 1153:1165 26 glycoforms of IgG1 1153:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 31 theme IgG 1193:1195 arg1 glycoforms 1209:1218 19 glycoforms 1206:1218 19 glycoforms of IgG4 1206:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 31 theme IgG 1193:1195 arg1 glycoforms 1179:1188 22 glycoforms 1176:1188 22 glycoforms of IgG 2/3 1176:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 32 theme unreported 1315:1324 arg1 glycoforms 1326:1335 previously unreported glycoforms 1304:1335 previously unreported glycoforms of IgG1 1304:1343 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 0 33 theme Unfractionated 58:71 arg1 Plasma 79:84 Unfractionated Human Plasma 58:84 Unfractionated Human Plasma 58:84 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 0 34 theme Data 0:3 arg1 Analysis 17:24 Data Independent Analysis 0:24 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 0:85 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 4 35 theme specific 557:564 arg1 glycoforms 566:575 site specific glycoforms 552:575 site specific glycoforms of IgG 552:582 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 5 36 theme ion 812:814 arg1 intensity 816:824 up to 60% precursor ion intensity 792:824 up to 60% precursor ion intensity 792:824 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 1 37 theme functional 111:120 arg1 responses 122:130 functional responses 111:130 functional responses mediated by the interaction of IgG with their receptors 111:186 Glycosylation regulates functional responses mediated by the interaction of IgG with their receptors. 27649061 0 38 theme IgG 29:31 arg1 Glycoforms 33:42 IgG Glycoforms 29:42 IgG Glycoforms 29:42 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 8 39 from changes 1535:1541 arg1 quantity 1550:1557 the quantity 1546:1557 the quantity of major glycoforms compared to healthy controls 1546:1606 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 9 40 from matrixes 1785:1792 arg1 analyses 1744:1751 quantitative analyses 1731:1751 quantitative analyses of the glycoproteome in complex matrixes 1731:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 3 41 theme abundant 400:407 arg1 glycoforms 409:418 low abundant glycoforms 396:418 low abundant glycoforms 396:418 However, measurement of low abundant glycoforms remains challenging in complex samples like serum without enrichment of the IgG. 27649061 4 42 theme quantitative 527:538 arg1 analysis 540:547 quantitative analysis 527:547 quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions 527:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 8 43 theme data 1397:1400 arg1 quantification 1414:1427 The SWATH data independent quantification 1387:1427 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis 1387:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 2 44 theme glycoforms 353:362 arg1 analysis 327:334 the analysis 323:334 the analysis of site specific glycoforms of IgG 323:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 6 45 theme selective 833:841 arg1 fragments 843:851 These selective fragments 827:851 These selective fragments 827:851 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 7 46 theme human 1267:1271 arg1 plasma 1273:1278 human plasma 1267:1278 human plasma 1267:1278 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 47 gly glycoforms 1179:1188 arg1 IgG4 1223:1226 IgG4 1223:1226 IgG4 1223:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 47 gly glycoforms 1179:1188 arg1 IgG 1193:1195 IgG 2/3 1193:1199 IgG 2/3 1193:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 47 gly glycoforms 1179:1188 arg1 IgG1 1170:1173 IgG1 1170:1173 IgG1 1170:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 2 48 theme site 339:342 arg1 glycoforms 353:362 site specific glycoforms 339:362 site specific glycoforms of IgG 339:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 2 49 theme analytical 198:207 arg1 methods 209:215 Multiple analytical methods 189:215 Multiple analytical methods 189:215 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 5 50 dep intensity 816:824 arg1 % 800:800 % 800:800 % 800:800 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 9 51 theme CID 1641:1643 arg1 fragmentation 1645:1657 optimized CID fragmentation 1631:1657 optimized CID fragmentation 1631:1657 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 8 52 theme glycoforms 1568:1577 arg1 quantity 1550:1557 the quantity 1546:1557 the quantity of major glycoforms compared to healthy controls 1546:1606 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 2 53 theme IgG 367:369 arg1 glycoforms 353:362 site specific glycoforms 339:362 site specific glycoforms of IgG 339:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 8 54 theme liver 1489:1493 arg1 cirrhosis 1495:1503 liver cirrhosis 1489:1503 liver cirrhosis 1489:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 7 55 theme reported 1125:1132 arg1 glycoforms 1134:1143 previously reported glycoforms 1114:1143 previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) 1114:1227 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 6 56 contain have 966:969 arg2 ratios 1022:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 6 56 contain have 966:969 arg1 B-ions 953:958 the typically quantified B-ions 928:958 the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios 928:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 2 57 theme MS 308:309 arg1 methods 311:317 MS methods 308:317 MS methods for the analysis of site specific glycoforms of IgG 308:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 5 58 dep % 800:800 arg1 60 798:799 60 798:799 60 798:799 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 4 59 theme data 593:596 arg1 DIA 623:625 DIA 623:625 DIA 623:625 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 4 59 theme data 593:596 arg1 acquisition 610:620 data independent acquisition 593:620 data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions 593:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 8 60 gly glycoforms 1436:1445 arg1 IgG 1432:1434 IgG glycoforms 1432:1445 IgG glycoforms 1432:1445 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 9 61 theme glycoforms 1678:1687 arg1 DIA 1667:1669 DIA 1667:1669 DIA of IgG glycoforms 1667:1687 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 6 62 theme higher 971:976 arg1 ratios 1022:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 4 63 theme " 662:662 arg1 conditions 678:687 "minimal" fragmentation conditions 654:687 "minimal" fragmentation conditions 654:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 5 64 theme collision 703:711 arg1 conditions 740:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 5 65 theme intensity 816:824 arg1 yield 783:787 the yield 779:787 the yield of up to 60% precursor ion intensity 779:824 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 7 66 gly glycoforms 1326:1335 arg1 IgG1 1340:1343 IgG1 1340:1343 IgG1 1340:1343 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 9 67 theme such 1706:1709 arg1 workflow 1711:1718 such workflow 1706:1718 such workflow 1706:1718 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 8 68 theme major 1562:1566 arg1 glycoforms 1568:1577 major glycoforms 1562:1577 major glycoforms compared to healthy controls 1562:1606 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 5 69 theme dissociation 721:732 arg1 conditions 740:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 7 70 gly fucosylated 1363:1373 arg1 glycoforms 1375:1384 doubly fucosylated glycoforms 1356:1384 doubly fucosylated glycoforms 1356:1384 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 6 71 theme quantified 942:951 arg1 B-ions 953:958 the typically quantified B-ions 928:958 the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios 928:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 8 72 theme plasma 1457:1462 arg1 samples 1464:1470 pooled plasma samples 1450:1470 pooled plasma samples of patients with liver cirrhosis 1450:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 4 73 gly glycoforms 566:575 arg1 IgG 580:582 IgG 580:582 IgG 580:582 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 9 74 theme quantitative 1731:1742 arg1 analyses 1744:1751 quantitative analyses 1731:1751 quantitative analyses of the glycoproteome in complex matrixes 1731:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 7 75 theme optimized 1036:1044 arg1 conditions 1046:1055 optimized conditions 1036:1055 optimized conditions 1036:1055 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 2 76 theme N-glycan 269:276 arg1 microheterogeneity 278:295 the IgG N-glycan microheterogeneity 261:295 the IgG N-glycan microheterogeneity 261:295 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 9 77 from analyses 1744:1751 arg1 matrixes 1785:1792 complex matrixes 1777:1792 complex matrixes 1777:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 6 78 dep higher 971:976 arg1 intensity 986:994 overall intensity 978:994 overall intensity 978:994 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 5 79 theme specific 760:767 arg1 Y-ions 769:774 specific Y-ions 760:774 specific Y-ions 760:774 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 9 80 theme glycoproteome 1760:1772 arg1 analyses 1744:1751 quantitative analyses 1731:1751 quantitative analyses of the glycoproteome in complex matrixes 1731:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 0 81 theme Plasma 79:84 arg1 Samples 47:53 Samples 47:53 Samples of Unfractionated Human Plasma 47:84 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 1 82 theme IgG 163:165 arg1 interaction 148:158 the interaction 144:158 the interaction of IgG with their receptors 144:186 Glycosylation regulates functional responses mediated by the interaction of IgG with their receptors. 27649061 6 83 theme lower 1000:1004 arg1 ratios 1022:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 higher overall intensity but lower signal-to-noise ratios 971:1027 These selective fragments, measured in high resolution, improve specificity of detection compared to the typically quantified B-ions which have higher overall intensity but lower signal-to-noise ratios. 27649061 9 84 theme complex 1777:1783 arg1 matrixes 1785:1792 complex matrixes 1777:1792 complex matrixes 1777:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 5 85 theme CID 735:737 arg1 conditions 740:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions 690:749 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 1 86 with interaction 148:158 arg1 receptors 178:186 their receptors 172:186 their receptors 172:186 Glycosylation regulates functional responses mediated by the interaction of IgG with their receptors. 27649061 8 87 theme expected 1526:1533 arg1 changes 1535:1541 the expected changes 1522:1541 the expected changes in the quantity of major glycoforms compared to healthy controls 1522:1606 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 0 88 theme Independent 5:15 arg1 Analysis 17:24 Data Independent Analysis 0:24 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 0:85 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 3 89 theme low 396:398 arg1 glycoforms 409:418 low abundant glycoforms 396:418 low abundant glycoforms 396:418 However, measurement of low abundant glycoforms remains challenging in complex samples like serum without enrichment of the IgG. 27649061 4 90 theme glycoforms 566:575 arg1 analysis 540:547 quantitative analysis 527:547 quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions 527:687 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 7 91 gly glycoforms 1209:1218 arg1 IgG4 1223:1226 IgG4 1223:1226 IgG4 1223:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 91 gly glycoforms 1209:1218 arg1 IgG 1193:1195 IgG 2/3 1193:1199 IgG 2/3 1193:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 91 gly glycoforms 1209:1218 arg1 IgG1 1170:1173 IgG1 1170:1173 IgG1 1170:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 3 92 theme complex 443:449 arg1 samples 451:457 complex samples 443:457 complex samples like serum without enrichment of the IgG 443:498 However, measurement of low abundant glycoforms remains challenging in complex samples like serum without enrichment of the IgG. 27649061 5 93 theme precursor 802:810 arg1 intensity 816:824 up to 60% precursor ion intensity 792:824 up to 60% precursor ion intensity 792:824 The adjusted collision induced dissociation (CID) conditions generate specific Y-ions in the yield of up to 60% precursor ion intensity. 27649061 8 94 theme IgG 1432:1434 arg1 glycoforms 1436:1445 IgG glycoforms 1432:1445 IgG glycoforms 1432:1445 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 4 95 theme IgG 580:582 arg1 glycoforms 566:575 site specific glycoforms 552:575 site specific glycoforms of IgG 552:582 We present a workflow for quantitative analysis of site specific glycoforms of IgG based on data independent acquisition (DIA) of Y-ions generated under "minimal" fragmentation conditions. 27649061 2 96 theme Multiple 189:196 arg1 methods 209:215 Multiple analytical methods 189:215 Multiple analytical methods 189:215 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 0 97 theme Glycoforms 33:42 arg1 Analysis 17:24 Data Independent Analysis 0:24 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 0:85 Data Independent Analysis of IgG Glycoforms in Samples of Unfractionated Human Plasma. 27649061 7 98 theme plasma 1273:1278 arg1 samples 1256:1262 unfractionated samples 1241:1262 unfractionated samples of human plasma 1241:1278 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 99 gly glycoforms 1156:1165 arg1 IgG4 1223:1226 IgG4 1223:1226 IgG4 1223:1226 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 99 gly glycoforms 1156:1165 arg1 IgG 1193:1195 IgG 2/3 1193:1199 IgG 2/3 1193:1199 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 7 99 gly glycoforms 1156:1165 arg1 IgG1 1170:1173 IgG1 1170:1173 IgG1 1170:1173 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 3 100 theme glycoforms 409:418 arg1 measurement 381:391 measurement 381:391 measurement of low abundant glycoforms 381:418 However, measurement of low abundant glycoforms remains challenging in complex samples like serum without enrichment of the IgG. 27649061 8 101 theme healthy 1591:1597 arg1 controls 1599:1606 healthy controls 1591:1606 healthy controls 1591:1606 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 2 102 gly glycoforms 353:362 arg1 IgG 367:369 IgG 367:369 IgG 367:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 2 102 gly glycoforms 353:362 arg1 site 339:342 site specific glycoforms 339:362 site specific glycoforms of IgG 339:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 8 103 theme SWATH 1391:1395 arg1 quantification 1414:1427 The SWATH data independent quantification 1387:1427 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis 1387:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 2 104 theme specific 344:351 arg1 glycoforms 353:362 site specific glycoforms 339:362 site specific glycoforms of IgG 339:369 Multiple analytical methods have been designed for the determination of the IgG N-glycan microheterogeneity, including MS methods for the analysis of site specific glycoforms of IgG. 27649061 7 105 theme unfractionated 1241:1254 arg1 samples 1256:1262 unfractionated samples 1241:1262 unfractionated samples of human plasma 1241:1278 Under optimized conditions, we achieve label-free quantification of the majority of previously reported glycoforms of IgG (26 glycoforms of IgG1, 22 glycoforms of IgG 2/3, and 19 glycoforms of IgG4) directly in unfractionated samples of human plasma and we detect traces of previously unreported glycoforms of IgG1, including doubly fucosylated glycoforms. 27649061 8 106 theme independent 1402:1412 arg1 quantification 1414:1427 The SWATH data independent quantification 1387:1427 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis 1387:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 3 107 theme IgG 496:498 arg1 enrichment 478:487 enrichment 478:487 enrichment of the IgG 478:498 However, measurement of low abundant glycoforms remains challenging in complex samples like serum without enrichment of the IgG. 27649061 9 108 from glycoproteome 1760:1772 arg1 matrixes 1785:1792 complex matrixes 1777:1792 complex matrixes 1777:1792 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 9 109 theme optimized 1631:1639 arg1 fragmentation 1645:1657 optimized CID fragmentation 1631:1657 optimized CID fragmentation 1631:1657 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 27649061 8 110 with patients 1475:1482 arg1 cirrhosis 1495:1503 liver cirrhosis 1489:1503 liver cirrhosis 1489:1503 The SWATH data independent quantification of IgG glycoforms in pooled plasma samples of patients with liver cirrhosis detects reliably the expected changes in the quantity of major glycoforms compared to healthy controls. 27649061 9 111 gly glycoforms 1678:1687 arg1 IgG 1674:1676 IgG glycoforms 1674:1687 IgG glycoforms 1674:1687 Our results show that optimized CID fragmentation enables DIA of IgG glycoforms and suggest that such workflow may enable quantitative analyses of the glycoproteome in complex matrixes. 28887379 13 0 theme fragmentation 2090:2102 arg1 necessary 2107:2115 necessary 2107:2115 necessary 2107:2115 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 13 0 theme fragmentation 2090:2102 arg1 consideration 2054:2066 the simultaneous consideration 2037:2066 the simultaneous consideration of peptide and glycan fragmentation 2037:2102 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 4 1 theme computational 633:645 arg1 platforms 647:655 comprehensive computational platforms 619:655 comprehensive computational platforms for data analyses 619:673 However, comprehensive computational platforms for data analyses are scarce. 28887379 4 1 theme computational 633:645 arg1 scarce 679:684 scarce 679:684 scarce 679:684 However, comprehensive computational platforms for data analyses are scarce. 28887379 3 2 theme high-throughput 527:541 arg1 workflow 561:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28887379 9 3 theme original 1586:1593 arg1 mass 1601:1604 the original sugar mass 1582:1604 the original sugar mass 1582:1604 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 3 4 theme site-specific 581:593 arg1 glycosylation 595:607 site-specific glycosylation 581:607 site-specific glycosylation 581:607 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28887379 9 5 theme decoy 1437:1441 arg1 glycopeptides 1443:1455 decoy glycopeptides 1437:1455 decoy glycopeptides 1437:1455 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 8 6 theme Ensemble 1214:1221 arg1 Score 1223:1227 the "Ensemble Score 1209:1227 the "Ensemble Score (ES) 1209:1232 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 8 6 theme Ensemble 1214:1221 arg1 ES 1230:1231 ES 1230:1231 ES 1230:1231 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 2 7 from nature 362:367 arg1 site 464:467 a given site 456:467 a given site 456:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 7 from nature 362:367 arg1 distribution 440:451 the glycan distribution 429:451 the glycan distribution 429:451 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 5 8 theme comprehensive 728:740 arg1 software 764:771 a comprehensive, open-source, modular software 726:771 a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat) 726:911 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 6 9 theme data 1033:1036 arg1 analysis 1038:1045 MSn data analysis 1029:1045 MSn data analysis 1029:1045 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 11 10 from mixtures 1807:1814 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 2 11 from glycans 372:378 arg1 distribution 440:451 the glycan distribution 429:451 the glycan distribution 429:451 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 10 12 theme Parallel 1607:1614 arg1 facilities 1626:1635 Parallel computing facilities 1607:1635 Parallel computing facilities 1607:1635 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 11 13 theme cryoprecipitate 1833:1847 arg1 samples 1849:1855 human plasma cryoprecipitate samples 1820:1855 human plasma cryoprecipitate samples 1820:1855 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 14 dep glycoproteins 1775:1787 arg1 ETD 1913:1915 ETD 1913:1915 ETD 1913:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 14 dep glycoproteins 1775:1787 arg1 glycoproteins 1775:1787 simple glycoproteins 1768:1787 simple glycoproteins 1768:1787 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 14 dep glycoproteins 1775:1787 arg1 HCD 1905:1907 HCD 1905:1907 HCD 1905:1907 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 14 dep glycoproteins 1775:1787 arg1 CID 1900:1902 CID 1900:1902 CID 1900:1902 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 12 15 theme cell 1974:1977 arg1 lysates 1979:1985 cell lysates 1974:1985 cell lysates from prostate cancer cells 1974:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 1 16 theme abundant 129:136 arg1 PTMs 192:195 PTMs 192:195 PTMs 192:195 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 1 16 theme abundant 129:136 arg1 modifications 177:189 the most abundant and diverse protein post-translational modifications 120:189 the most abundant and diverse protein post-translational modifications (PTMs) identified to date 120:215 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 5 17 dep GlycoPAT 814:821 arg1 Toolbox 849:855 GlycoProteomics Analysis Toolbox 824:855 GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat 824:910 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 11 18 theme fragmentation 1879:1891 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 7 19 theme PTM 1128:1130 arg1 fragmentation 1081:1093 facile serial fragmentation 1067:1093 facile serial fragmentation of both the peptide backbone and PTM 1067:1130 This format allows facile serial fragmentation of both the peptide backbone and PTM at one or more locations. 28887379 11 20 from glycoproteins 1775:1787 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 9 21 gly glycopeptides 1443:1455 arg2 glycopeptides 1443:1455 decoy glycopeptides 1437:1455 decoy glycopeptides 1437:1455 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 10 22 theme user-friendly 1641:1653 arg1 GUIs 1655:1658 user-friendly GUIs 1641:1658 user-friendly GUIs (Graphical User Interfaces) 1641:1686 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 10 22 theme user-friendly 1641:1653 arg1 Interfaces 1676:1685 Graphical User Interfaces 1661:1685 Graphical User Interfaces 1661:1685 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 11 23 theme common 1866:1871 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 7 24 theme backbone 1115:1122 arg1 fragmentation 1081:1093 facile serial fragmentation 1067:1093 facile serial fragmentation of both the peptide backbone and PTM 1067:1130 This format allows facile serial fragmentation of both the peptide backbone and PTM at one or more locations. 28887379 13 25 theme quality 2126:2132 arg1 annotation 2147:2156 high quality MSn spectrum annotation 2121:2156 high quality MSn spectrum annotation in CID and HCD fragmentation modes 2121:2191 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 7 26 theme facile 1067:1072 arg1 fragmentation 1081:1093 facile serial fragmentation 1067:1093 facile serial fragmentation of both the peptide backbone and PTM 1067:1130 This format allows facile serial fragmentation of both the peptide backbone and PTM at one or more locations. 28887379 9 27 theme amino 1502:1506 arg1 sequence 1513:1520 the amino acid sequence 1498:1520 the amino acid sequence 1498:1520 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 6 28 theme linear 986:991 arg1 representation 993:1006 a minimal linear representation 976:1006 a minimal linear representation of glycopeptides for MSn data analysis 976:1045 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 11 29 gly glycosylation 1751:1763 arg1 CID 1900:1902 CID 1900:1902 CID 1900:1902 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 29 gly glycosylation 1751:1763 arg1 mixtures 1807:1814 standard protein mixtures 1790:1814 standard protein mixtures 1790:1814 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 29 gly glycosylation 1751:1763 arg1 HCD 1905:1907 HCD 1905:1907 HCD 1905:1907 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 29 gly glycosylation 1751:1763 arg1 glycoproteins 1775:1787 simple glycoproteins 1768:1787 simple glycoproteins 1768:1787 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 29 gly glycosylation 1751:1763 arg1 ETD 1913:1915 ETD 1913:1915 ETD 1913:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 29 gly glycosylation 1751:1763 arg1 samples 1849:1855 human plasma cryoprecipitate samples 1820:1855 human plasma cryoprecipitate samples 1820:1855 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 30 theme standard 1790:1797 arg1 mixtures 1807:1814 standard protein mixtures 1790:1814 standard protein mixtures 1790:1814 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 13 31 theme spectrum 2138:2145 arg1 annotation 2147:2156 high quality MSn spectrum annotation 2121:2156 high quality MSn spectrum annotation in CID and HCD fragmentation modes 2121:2191 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 1 32 theme diverse 142:148 arg1 PTMs 192:195 PTMs 192:195 PTMs 192:195 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 1 32 theme diverse 142:148 arg1 modifications 177:189 the most abundant and diverse protein post-translational modifications 120:189 the most abundant and diverse protein post-translational modifications (PTMs) identified to date 120:215 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 8 33 theme O-linked 1300:1307 arg1 glycopeptides 1309:1321 O-linked glycopeptides 1300:1321 O-linked glycopeptides using cross-correlation and probability based analyses 1300:1376 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 12 34 gly glycopeptides 1957:1969 arg2 glycopeptides 1957:1969 960 unique glycopeptides 1946:1969 960 unique glycopeptides in cell lysates from prostate cancer cells 1946:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 0 35 theme Glycoproteomics 66:80 arg1 Analysis 87:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis. 28887379 1 36 theme post-translational 158:175 arg1 PTMs 192:195 PTMs 192:195 PTMs 192:195 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 1 36 theme post-translational 158:175 arg1 modifications 177:189 the most abundant and diverse protein post-translational modifications 120:189 the most abundant and diverse protein post-translational modifications (PTMs) identified to date 120:215 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 14 37 theme GlycoPAT 2240:2247 arg1 suitability 2225:2235 the suitability 2221:2235 the suitability of GlycoPAT to analyze shotgun glycoproteomics data 2221:2287 Additionally, they confirm the suitability of GlycoPAT to analyze shotgun glycoproteomics data. 28887379 11 38 theme human 1820:1824 arg1 samples 1849:1855 human plasma cryoprecipitate samples 1820:1855 human plasma cryoprecipitate samples 1820:1855 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 5 39 theme Analysis 840:847 arg1 Toolbox 849:855 GlycoProteomics Analysis Toolbox 824:855 GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat 824:910 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 9 40 theme artificial 1541:1550 arg1 monosaccharides 1552:1566 artificial monosaccharides 1541:1566 artificial monosaccharides 1541:1566 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 8 41 theme based 1363:1367 arg1 analyses 1369:1376 probability based analyses 1351:1376 probability based analyses 1351:1376 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 5 42 theme data 793:796 arg1 analysis 798:805 glycoproteomics data analysis 777:805 glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat) 777:911 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 8 43 gly glycopeptides 1309:1321 arg2 glycopeptides 1309:1321 O-linked glycopeptides 1300:1321 O-linked glycopeptides using cross-correlation and probability based analyses 1300:1376 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 12 44 theme unique 1950:1955 arg1 glycopeptides 1957:1969 960 unique glycopeptides 1946:1969 960 unique glycopeptides in cell lysates from prostate cancer cells 1946:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 8 45 dep based 1185:1189 arg1 2 1159:1159 2 1159:1159 2 1159:1159 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 13 46 theme glycan 2083:2088 arg1 fragmentation 2090:2102 glycan fragmentation 2083:2102 glycan fragmentation 2083:2102 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 2 47 theme isomeric 387:394 arg1 structures 396:405 their isomeric structures 381:405 their isomeric structures 381:405 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 9 48 theme sugar 1595:1599 arg1 mass 1601:1604 the original sugar mass 1582:1604 the original sugar mass 1582:1604 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 2 49 theme heterogeneity 412:424 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 49 theme heterogeneity 412:424 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 14 50 theme glycoproteomics 2268:2282 arg1 data 2284:2287 shotgun glycoproteomics data 2260:2287 shotgun glycoproteomics data 2260:2287 Additionally, they confirm the suitability of GlycoPAT to analyze shotgun glycoproteomics data. 28887379 13 51 theme simultaneous 2041:2052 arg1 necessary 2107:2115 necessary 2107:2115 necessary 2107:2115 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 13 51 theme simultaneous 2041:2052 arg1 consideration 2054:2066 the simultaneous consideration 2037:2066 the simultaneous consideration of peptide and glycan fragmentation 2037:2102 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 2 52 theme branched 353:360 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 52 theme branched 353:360 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 5 53 theme available 865:873 arg1 Toolbox 849:855 GlycoProteomics Analysis Toolbox 824:855 GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat 824:910 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 11 54 theme simple 1768:1773 arg1 ETD 1913:1915 ETD 1913:1915 ETD 1913:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 54 theme simple 1768:1773 arg1 glycoproteins 1775:1787 simple glycoproteins 1768:1787 simple glycoproteins 1768:1787 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 54 theme simple 1768:1773 arg1 HCD 1905:1907 HCD 1905:1907 HCD 1905:1907 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 54 theme simple 1768:1773 arg1 CID 1900:1902 CID 1900:1902 CID 1900:1902 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 9 55 theme false 1385:1389 arg1 rate 1401:1404 A false discovery rate 1383:1404 A false discovery rate (FDR) 1383:1410 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 9 55 theme false 1385:1389 arg1 FDR 1407:1409 FDR 1407:1409 FDR 1407:1409 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 2 56 from heterogeneity 412:424 arg1 distribution 440:451 the glycan distribution 429:451 the glycan distribution 429:451 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 6 57 dep 1 958:958 arg1 SmallGlyPep 962:972 SmallGlyPep 962:972 three major advances: (1) "SmallGlyPep 935:972 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 9 58 dep created 1461:1467 arg1 introducing 1529:1539 introducing 1529:1539 introducing artificial monosaccharides by perturbing the original sugar mass 1529:1604 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 9 58 dep created 1461:1467 arg1 scrambling 1487:1496 scrambling 1487:1496 scrambling the amino acid sequence 1487:1520 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 3 59 theme LC-MSn 543:548 arg1 workflow 561:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28887379 8 60 theme novel 1164:1168 arg1 scheme 1178:1183 A novel scoring scheme 1162:1183 A novel scoring scheme 1162:1183 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 6 61 theme minimal 978:984 arg1 representation 993:1006 a minimal linear representation 976:1006 a minimal linear representation of glycopeptides for MSn data analysis 976:1045 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 12 62 theme cancer 2001:2006 arg1 cells 2008:2012 prostate cancer cells 1992:2012 prostate cancer cells 1992:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 13 63 dep CID 2161:2163 arg1 modes 2187:2191 modes 2187:2191 modes 2187:2191 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 14 64 theme shotgun 2260:2266 arg1 data 2284:2287 shotgun glycoproteomics data 2260:2287 shotgun glycoproteomics data 2260:2287 Additionally, they confirm the suitability of GlycoPAT to analyze shotgun glycoproteomics data. 28887379 3 65 theme traditional 515:525 arg1 workflow 561:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28887379 4 66 theme comprehensive 619:631 arg1 platforms 647:655 comprehensive computational platforms 619:655 comprehensive computational platforms for data analyses 619:673 However, comprehensive computational platforms for data analyses are scarce. 28887379 4 66 theme comprehensive 619:631 arg1 scarce 679:684 scarce 679:684 scarce 679:684 However, comprehensive computational platforms for data analyses are scarce. 28887379 6 67 gly glycopeptides 1011:1023 arg2 glycopeptides 1011:1023 glycopeptides 1011:1023 glycopeptides 1011:1023 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 2 68 theme diverse 284:290 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 68 theme diverse 284:290 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 11 69 from samples 1849:1855 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 12 70 theme prostate 1992:1999 arg1 cells 2008:2012 prostate cancer cells 1992:2012 prostate cancer cells 1992:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 10 71 theme computing 1616:1624 arg1 facilities 1626:1635 Parallel computing facilities 1607:1635 Parallel computing facilities 1607:1635 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 8 72 theme Score 1223:1227 arg1 calculation 1194:1204 calculation 1194:1204 calculation of the "Ensemble Score (ES) 1194:1232 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 6 73 theme MSn 1029:1031 arg1 analysis 1038:1045 MSn data analysis 1029:1045 MSn data analysis 1029:1045 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 10 74 theme Graphical 1661:1669 arg1 GUIs 1655:1658 user-friendly GUIs 1641:1658 user-friendly GUIs (Graphical User Interfaces) 1641:1686 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 10 74 theme Graphical 1661:1669 arg1 Interfaces 1676:1685 Graphical User Interfaces 1661:1685 Graphical User Interfaces 1661:1685 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 2 75 theme PTM 250:252 arg1 analysis 233:240 The structural analysis 218:240 The structural analysis of this PTM 218:252 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 6 76 theme glycopeptides 1011:1023 arg1 representation 993:1006 a minimal linear representation 976:1006 a minimal linear representation of glycopeptides for MSn data analysis 976:1045 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 8 77 theme MS/MS 1274:1278 arg1 spectrum 1280:1287 MS/MS spectrum 1274:1287 MS/MS spectrum 1274:1287 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 12 78 used used 1929:1932 arg2 It 1918:1919 It 1918:1919 It 1918:1919 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 7 79 theme peptide 1107:1113 arg1 backbone 1115:1122 the peptide backbone 1103:1122 the peptide backbone 1103:1122 This format allows facile serial fragmentation of both the peptide backbone and PTM at one or more locations. 28887379 13 80 theme HCD 2169:2171 arg1 fragmentation 2173:2185 HCD fragmentation 2169:2185 HCD fragmentation 2169:2185 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 9 81 dep calculation 1412:1422 arg1 3 1380:1380 3 1380:1380 3 1380:1380 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 11 82 theme MS/MS 1873:1877 arg1 modes 1893:1897 three common MS/MS fragmentation modes 1860:1897 three common MS/MS fragmentation modes 1860:1897 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 13 83 theme MSn 2134:2136 arg1 annotation 2147:2156 high quality MSn spectrum annotation 2121:2156 high quality MSn spectrum annotation in CID and HCD fragmentation modes 2121:2191 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 7 84 theme serial 1074:1079 arg1 fragmentation 1081:1093 facile serial fragmentation 1067:1093 facile serial fragmentation of both the peptide backbone and PTM 1067:1130 This format allows facile serial fragmentation of both the peptide backbone and PTM at one or more locations. 28887379 9 85 theme acid 1508:1511 arg1 sequence 1513:1520 the amino acid sequence 1498:1520 the amino acid sequence 1498:1520 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 12 86 from glycopeptides 1957:1969 arg1 lysates 1979:1985 cell lysates 1974:1985 cell lysates from prostate cancer cells 1974:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 12 86 from glycopeptides 1957:1969 arg1 cells 2008:2012 prostate cancer cells 1992:2012 prostate cancer cells 1992:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 11 87 theme protein 1799:1805 arg1 mixtures 1807:1814 standard protein mixtures 1790:1814 standard protein mixtures 1790:1814 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 0 88 theme Data 82:85 arg1 Analysis 87:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis. 28887379 1 89 theme protein 150:156 arg1 PTMs 192:195 PTMs 192:195 PTMs 192:195 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 1 89 theme protein 150:156 arg1 modifications 177:189 the most abundant and diverse protein post-translational modifications 120:189 the most abundant and diverse protein post-translational modifications (PTMs) identified to date 120:215 Glycosylation is among the most abundant and diverse protein post-translational modifications (PTMs) identified to date. 28887379 11 90 from glycosylation 1751:1763 arg1 CID 1900:1902 CID 1900:1902 CID 1900:1902 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 90 from glycosylation 1751:1763 arg1 mixtures 1807:1814 standard protein mixtures 1790:1814 standard protein mixtures 1790:1814 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 90 from glycosylation 1751:1763 arg1 HCD 1905:1907 HCD 1905:1907 HCD 1905:1907 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 90 from glycosylation 1751:1763 arg1 glycoproteins 1775:1787 simple glycoproteins 1768:1787 simple glycoproteins 1768:1787 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 90 from glycosylation 1751:1763 arg1 ETD 1913:1915 ETD 1913:1915 ETD 1913:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 90 from glycosylation 1751:1763 arg1 samples 1849:1855 human plasma cryoprecipitate samples 1820:1855 human plasma cryoprecipitate samples 1820:1855 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 2 91 from distribution 440:451 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 91 from distribution 440:451 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 5 92 theme GlycoProteomics 824:838 arg1 Toolbox 849:855 GlycoProteomics Analysis Toolbox 824:855 GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat 824:910 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 13 93 theme peptide 2071:2077 arg1 necessary 2107:2115 necessary 2107:2115 necessary 2107:2115 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 13 93 theme peptide 2071:2077 arg1 consideration 2054:2066 the simultaneous consideration 2037:2066 the simultaneous consideration of peptide and glycan fragmentation 2037:2102 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 11 94 theme plasma 1826:1831 arg1 samples 1849:1855 human plasma cryoprecipitate samples 1820:1855 human plasma cryoprecipitate samples 1820:1855 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 5 95 theme glycoproteomics 777:791 arg1 analysis 798:805 glycoproteomics data analysis 777:805 glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat) 777:911 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 13 96 theme high 2121:2124 arg1 annotation 2147:2156 high quality MSn spectrum annotation 2121:2156 high quality MSn spectrum annotation in CID and HCD fragmentation modes 2121:2191 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 2 97 from site 464:467 arg1 structures 396:405 their isomeric structures 381:405 their isomeric structures 381:405 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 97 from site 464:467 arg1 heterogeneity 412:424 heterogeneity 412:424 heterogeneity in the glycan distribution at a given site 412:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 97 from site 464:467 arg1 glycans 372:378 glycans 372:378 glycans 372:378 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 97 from site 464:467 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 97 from site 464:467 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 8 98 theme probability 1351:1361 arg1 analyses 1369:1376 probability based analyses 1351:1376 probability based analyses 1351:1376 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 5 99 dep comprehensive 728:740 arg1 modular 756:762 modular 756:762 modular 756:762 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 5 99 dep comprehensive 728:740 arg1 open-source 743:753 open-source 743:753 open-source 743:753 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 4 100 theme data 661:664 arg1 analyses 666:673 data analyses 661:673 data analyses 661:673 However, comprehensive computational platforms for data analyses are scarce. 28887379 2 101 theme structures 396:405 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 101 theme structures 396:405 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 0 102 theme Comprehensive 2:14 arg1 Platform 29:36 A Comprehensive, Open-source Platform 0:36 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis 0:94 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis. 28887379 5 103 from www.VirtualGlycome.org/glycopat 880:910 arg1 available 865:873 available 865:873 available 865:873 To address this limitation, we present a comprehensive, open-source, modular software for glycoproteomics data analysis called GlycoPAT (GlycoProteomics Analysis Toolbox; freely available from www.VirtualGlycome.org/glycopat). 28887379 11 104 used used 1719:1722 arg2 GlycoPAT 1707:1714 GlycoPAT 1707:1714 GlycoPAT 1707:1714 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 8 105 dep scores 1251:1256 arg1 spectrum 1280:1287 MS/MS spectrum 1274:1287 MS/MS spectrum 1274:1287 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 10 106 theme User 1671:1674 arg1 GUIs 1655:1658 user-friendly GUIs 1641:1658 user-friendly GUIs (Graphical User Interfaces) 1641:1686 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 10 106 theme User 1671:1674 arg1 Interfaces 1676:1685 Graphical User Interfaces 1661:1685 Graphical User Interfaces 1661:1685 Parallel computing facilities and user-friendly GUIs (Graphical User Interfaces) are also provided. 28887379 2 107 theme glycan 433:438 arg1 distribution 440:451 the glycan distribution 429:451 the glycan distribution 429:451 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 11 108 gly glycoproteins 1775:1787 arg1 ETD 1913:1915 ETD 1913:1915 ETD 1913:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 108 gly glycoproteins 1775:1787 arg1 glycoproteins 1775:1787 simple glycoproteins 1768:1787 simple glycoproteins 1768:1787 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 108 gly glycoproteins 1775:1787 arg1 HCD 1905:1907 HCD 1905:1907 HCD 1905:1907 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 11 108 gly glycoproteins 1775:1787 arg1 CID 1900:1902 CID 1900:1902 CID 1900:1902 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 3 109 theme Glycoproteomics 470:484 arg1 experiments 486:496 Glycoproteomics experiments 470:496 Glycoproteomics experiments 470:496 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28887379 12 110 from cells 2008:2012 arg1 lysates 1979:1985 cell lysates 1974:1985 cell lysates from prostate cancer cells 1974:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 12 110 from cells 2008:2012 arg1 glycopeptides 1957:1969 960 unique glycopeptides 1946:1969 960 unique glycopeptides in cell lysates from prostate cancer cells 1946:2012 It is also used to identify 960 unique glycopeptides in cell lysates from prostate cancer cells. 28887379 6 111 dep advances 947:954 arg1 1 958:958 1 958:958 1 958:958 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 0 112 theme Spectrometry-based 47:64 arg1 Analysis 87:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 Mass Spectrometry-based Glycoproteomics Data Analysis 42:94 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis. 28887379 6 113 theme major 941:945 arg1 advances 947:954 three major advances 935:954 three major advances: (1) "SmallGlyPep 935:972 The program includes three major advances: (1) "SmallGlyPep," a minimal linear representation of glycopeptides for MSn data analysis. 28887379 2 114 theme given 458:462 arg1 site 464:467 a given site 456:467 a given site 456:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 8 115 link O-linked 1300:1307 arg1 glycopeptides 1309:1321 O-linked glycopeptides 1300:1321 O-linked glycopeptides using cross-correlation and probability based analyses 1300:1376 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 2 116 theme structural 222:231 arg1 analysis 233:240 The structural analysis 218:240 The structural analysis of this PTM 218:252 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 117 from structures 396:405 arg1 distribution 440:451 the glycan distribution 429:451 the glycan distribution 429:451 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 11 118 theme site-specific 1737:1749 arg1 glycosylation 1751:1763 site-specific glycosylation 1737:1763 site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD 1737:1915 GlycoPAT is used to catalogue site-specific glycosylation on simple glycoproteins, standard protein mixtures and human plasma cryoprecipitate samples in three common MS/MS fragmentation modes: CID, HCD and ETD. 28887379 2 119 theme glycans 372:378 arg1 monosaccharides 292:306 the diverse monosaccharides 280:306 the diverse monosaccharides which are not conserved among organisms 280:346 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 2 119 theme glycans 372:378 arg1 nature 362:367 the branched nature 349:367 the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site 349:467 The structural analysis of this PTM is challenging because of the diverse monosaccharides which are not conserved among organisms, the branched nature of glycans, their isomeric structures, and heterogeneity in the glycan distribution at a given site. 28887379 9 120 theme discovery 1391:1399 arg1 rate 1401:1404 A false discovery rate 1383:1404 A false discovery rate (FDR) 1383:1410 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 9 120 theme discovery 1391:1399 arg1 FDR 1407:1409 FDR 1407:1409 FDR 1407:1409 (3) A false discovery rate (FDR) calculation scheme where decoy glycopeptides are created by simultaneously scrambling the amino acid sequence and by introducing artificial monosaccharides by perturbing the original sugar mass. 28887379 8 121 theme scoring 1170:1176 arg1 scheme 1178:1183 A novel scoring scheme 1162:1183 A novel scoring scheme 1162:1183 (2) A novel scoring scheme based on calculation of the "Ensemble Score (ES)," a measure that scores and rank-orders MS/MS spectrum for N- and O-linked glycopeptides using cross-correlation and probability based analyses. 28887379 0 122 dep Comprehensive 2:14 arg1 Open-source 17:27 Open-source 17:27 Open-source 17:27 A Comprehensive, Open-source Platform for Mass Spectrometry-based Glycoproteomics Data Analysis. 28887379 13 123 from annotation 2147:2156 arg1 CID 2161:2163 CID 2161:2163 CID 2161:2163 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 13 123 from annotation 2147:2156 arg1 fragmentation 2173:2185 HCD fragmentation 2169:2185 HCD fragmentation 2169:2185 The results show that the simultaneous consideration of peptide and glycan fragmentation is necessary for high quality MSn spectrum annotation in CID and HCD fragmentation modes. 28887379 3 124 theme proteomics 550:559 arg1 workflow 561:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 the traditional high-throughput LC-MSn proteomics workflow 511:568 Glycoproteomics experiments have adopted the traditional high-throughput LC-MSn proteomics workflow to analyze site-specific glycosylation. 28935113 6 0 theme desired 1041:1047 arg1 glycan 1049:1054 any desired glycan 1037:1054 any desired glycan 1037:1054 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 7 1 theme in 1077:1078 arg1 selection 1094:1102 in vitro peptide selection 1077:1102 in vitro peptide selection 1077:1102 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 9 2 theme high-mannose 1519:1530 arg1 oligosaccharides 1532:1547 high-mannose oligosaccharides 1519:1547 high-mannose oligosaccharides on the HIV envelope glycoprotein gp120 1519:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 7 3 theme oligosaccharides 1197:1212 arg1 incorporation 1146:1158 unnatural amino acid incorporation 1125:1158 unnatural amino acid incorporation 1125:1158 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 7 3 theme oligosaccharides 1197:1212 arg1 attachment 1173:1182 chemical attachment 1164:1182 chemical attachment 1164:1182 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 4 4 from site 656:659 arg1 protein 668:674 the protein 664:674 the protein 664:674 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 2 5 theme glycosylation 245:257 arg1 importance 231:240 importance 231:240 importance 231:240 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 2 5 theme glycosylation 245:257 arg1 abundance 217:225 abundance 217:225 abundance 217:225 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 1 6 theme desired 162:168 arg1 abilities 178:186 desired binding abilities 162:186 desired binding abilities 162:186 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 9 7 theme vaccine 1652:1658 arg1 applications 1660:1671 HIV vaccine applications 1648:1671 HIV vaccine applications 1648:1671 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 4 8 theme heterogeneous 612:624 arg1 mixture 626:632 a heterogeneous mixture 610:632 a heterogeneous mixture of glycoforms 610:646 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 7 9 dep in 1077:1078 arg1 vitro 1080:1084 vitro 1080:1084 vitro 1080:1084 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 2 10 theme directed 270:277 arg1 evolution 279:287 directed evolution 270:287 directed evolution of glycopeptides and glycoproteins 270:322 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 9 11 from useful 1638:1643 arg1 applications 1660:1671 HIV vaccine applications 1648:1671 HIV vaccine applications 1648:1671 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 8 12 theme updated 1235:1241 arg1 protocol 1257:1264 an updated and optimized protocol 1232:1264 an updated and optimized protocol 1232:1264 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 5 13 theme glycosylation 789:801 arg1 sites 803:807 all glycosylation sites 785:807 all glycosylation sites 785:807 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 8 14 theme optimized 1247:1255 arg1 protocol 1257:1264 an updated and optimized protocol 1232:1264 an updated and optimized protocol 1232:1264 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 2 15 theme importance 231:240 arg1 desirable 339:347 desirable 339:347 desirable 339:347 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 9 16 theme monoclonal 1468:1477 arg1 antibody 1479:1486 monoclonal antibody 2G12 1468:1491 monoclonal antibody 2G12 1468:1491 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 6 17 theme glycopeptide 970:981 arg1 libraries 983:991 glycopeptide libraries 970:991 glycopeptide libraries 970:991 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 7 18 theme synthetic 1187:1195 arg1 oligosaccharides 1197:1212 synthetic oligosaccharides 1187:1212 synthetic oligosaccharides 1187:1212 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 2 19 theme abundance 217:225 arg1 desirable 339:347 desirable 339:347 desirable 339:347 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 1 20 theme peptides 130:137 arg1 discovery 102:110 the discovery 98:110 the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions 98:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 9 21 from cluster 1508:1514 arg1 gp120 1582:1586 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 5 22 theme Escherichia 707:717 arg1 cells 724:728 Escherichia coli cells 707:728 Escherichia coli cells 707:728 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 10 23 from targets 1754:1760 arg1 glycobiology 1777:1788 glycobiology 1777:1788 glycobiology 1777:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 9 24 from applications 1660:1671 arg1 useful 1638:1643 useful 1638:1643 useful 1638:1643 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 7 25 theme acid 1141:1144 arg1 incorporation 1146:1158 unnatural amino acid incorporation 1125:1158 unnatural amino acid incorporation 1125:1158 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 10 26 from interest 1765:1772 arg1 glycobiology 1777:1788 glycobiology 1777:1788 glycobiology 1777:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 3 27 theme directed 366:373 arg1 display 437:443 mammalian-cell display 422:443 mammalian-cell display 422:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 27 theme directed 366:373 arg1 yeast- 411:416 yeast- 411:416 yeast- 411:416 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 27 theme directed 366:373 arg1 platforms 385:393 common directed evolution platforms 359:393 common directed evolution platforms such as phage-, yeast-, or mammalian-cell display 359:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 27 theme directed 366:373 arg1 phage- 403:408 phage- 403:408 phage- 403:408 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 1 28 theme acids 123:127 arg1 discovery 102:110 the discovery 98:110 the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions 98:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 8 29 theme mRNA 1324:1327 arg1 libraries 1337:1345 glycopeptide mRNA display libraries 1311:1345 glycopeptide mRNA display libraries containing ~1013 sequences 1311:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 7 30 theme unnatural 1125:1133 arg1 incorporation 1146:1158 unnatural amino acid incorporation 1125:1158 unnatural amino acid incorporation 1125:1158 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 9 31 theme oligosaccharides 1532:1547 arg1 cluster 1508:1514 cluster 1508:1514 cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120 1508:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 10 32 from glycobiology 1777:1788 arg1 types 1733:1737 other types 1727:1737 other types of glycans 1727:1748 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 10 32 from glycobiology 1777:1788 arg1 targets 1754:1760 targets 1754:1760 targets of interest in glycobiology 1754:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 4 33 from site 539:542 arg1 structure 507:515 Glycan structure 500:515 Glycan structure at each glycosylation site 500:542 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 1 34 contain have 157:160 arg2 functions 191:199 functions 191:199 functions 191:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 34 contain have 157:160 arg1 peptides 130:137 peptides 130:137 peptides 130:137 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 34 contain have 157:160 arg1 proteins 143:150 proteins 143:150 proteins 143:150 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 34 contain have 157:160 arg1 acids 123:127 nucleic acids 115:127 nucleic acids 115:127 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 34 contain have 157:160 arg2 abilities 178:186 desired binding abilities 162:186 desired binding abilities 162:186 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 8 35 theme ~1013 1358:1362 arg1 sequences 1364:1372 ~1013 sequences 1358:1372 ~1013 sequences 1358:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 3 36 theme mammalian-cell 422:435 arg1 display 437:443 mammalian-cell display 422:443 mammalian-cell display 422:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 8 37 gly glycopeptide 1311:1322 arg2 glycopeptide 1311:1322 glycopeptide mRNA display libraries 1311:1345 glycopeptide mRNA display libraries containing ~1013 sequences 1311:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 9 38 theme envelope 1560:1567 arg1 gp120 1582:1586 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 8 39 contain containing 1347:1356 arg2 sequences 1364:1372 ~1013 sequences 1358:1372 ~1013 sequences 1358:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 8 39 contain containing 1347:1356 arg1 libraries 1337:1345 glycopeptide mRNA display libraries 1311:1345 glycopeptide mRNA display libraries containing ~1013 sequences 1311:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 7 40 theme peptide 1086:1092 arg1 selection 1094:1102 in vitro peptide selection 1077:1102 in vitro peptide selection 1077:1102 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 2 41 theme glycoproteins 310:322 arg1 evolution 279:287 directed evolution 270:287 directed evolution of glycopeptides and glycoproteins 270:322 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 8 42 theme target 1394:1399 arg1 binding 1401:1407 target binding 1394:1407 target binding 1394:1407 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 10 43 from types 1733:1737 arg1 glycobiology 1777:1788 glycobiology 1777:1788 glycobiology 1777:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 4 44 theme glycoforms 637:646 arg1 mixture 626:632 a heterogeneous mixture 610:632 a heterogeneous mixture of glycoforms 610:646 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 9 45 gly glycoprotein 1569:1580 arg1 glycoprotein 1569:1580 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 3 46 theme several 483:489 arg1 factors 491:497 several factors 483:497 several factors 483:497 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 10 47 theme glycans 1742:1748 arg1 types 1733:1737 other types 1727:1737 other types of glycans 1727:1748 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 10 47 theme glycans 1742:1748 arg1 targets 1754:1760 targets 1754:1760 targets of interest in glycobiology 1754:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 9 48 gly glycopeptides 1593:1605 arg2 glycopeptides 1593:1605 glycopeptides 1593:1605 glycopeptides that mimic this epitope 1593:1629 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 1 49 theme proteins 143:150 arg1 discovery 102:110 the discovery 98:110 the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions 98:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 2 50 dep abundance 217:225 arg1 the 213:215 the 213:215 the 213:215 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 2 51 theme glycopeptides 292:304 arg1 evolution 279:287 directed evolution 270:287 directed evolution of glycopeptides and glycoproteins 270:322 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 5 52 theme specific 831:838 arg1 structures 847:856 just a few specific glycan structures 820:856 just a few specific glycan structures that can readily be accessed in this manner 820:900 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 6 53 theme novel 927:931 arg1 system 933:938 a novel system 925:938 a novel system 925:938 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 9 54 theme HIV 1648:1650 arg1 applications 1660:1671 HIV vaccine applications 1648:1671 HIV vaccine applications 1648:1671 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 7 55 with display 1112:1118 arg1 incorporation 1146:1158 unnatural amino acid incorporation 1125:1158 unnatural amino acid incorporation 1125:1158 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 7 55 with display 1112:1118 arg1 attachment 1173:1182 chemical attachment 1164:1182 chemical attachment 1164:1182 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 1 56 theme binding 170:176 arg1 abilities 178:186 desired binding abilities 162:186 desired binding abilities 162:186 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 10 57 theme interest 1765:1772 arg1 types 1733:1737 other types 1727:1737 other types of glycans 1727:1748 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 10 57 theme interest 1765:1772 arg1 targets 1754:1760 targets 1754:1760 targets of interest in glycobiology 1754:1788 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 5 58 theme glycan 840:845 arg1 structures 847:856 just a few specific glycan structures 820:856 just a few specific glycan structures that can readily be accessed in this manner 820:900 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 9 59 from gp120 1582:1586 arg1 cluster 1508:1514 cluster 1508:1514 cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120 1508:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 7 60 theme mRNA 1107:1110 arg1 display 1112:1118 mRNA display 1107:1118 mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides 1107:1212 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 1 61 theme nucleic 115:121 arg1 acids 123:127 nucleic acids 115:127 nucleic acids 115:127 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 6 62 theme libraries 983:991 arg1 evolution 957:965 the directed evolution 944:965 the directed evolution of glycopeptide libraries 944:991 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 5 63 dep Escherichia 707:717 arg1 coli 719:722 coli 719:722 coli 719:722 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 9 64 from oligosaccharides 1532:1547 arg1 gp120 1582:1586 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 7 65 theme chemical 1164:1171 arg1 attachment 1173:1182 chemical attachment 1164:1182 chemical attachment 1164:1182 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 4 66 gly glycosylation 525:537 arg2 site 539:542 each glycosylation site 520:542 each glycosylation site 520:542 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 6 67 theme directed 948:955 arg1 evolution 957:965 the directed evolution 944:965 the directed evolution of glycopeptide libraries 944:991 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 4 68 theme Glycan 500:505 arg1 structure 507:515 Glycan structure 500:515 Glycan structure at each glycosylation site 500:542 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 1 69 theme Directed 56:63 arg1 method 87:92 a useful method 78:92 a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions 78:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 69 theme Directed 56:63 arg1 evolution 65:73 Directed evolution 56:73 Directed evolution 56:73 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 10 70 theme other 1727:1731 arg1 types 1733:1737 other types 1727:1737 other types of glycans 1727:1748 This method is expected to be readily applicable for other types of glycans and targets of interest in glycobiology. 28935113 0 71 theme Directed 0:7 arg1 Evolution 9:17 Directed Evolution 0:17 Directed Evolution of Glycopeptides 0:34 Directed Evolution of Glycopeptides Using mRNA Display. 28935113 7 72 theme amino 1135:1139 arg1 incorporation 1146:1158 unnatural amino acid incorporation 1125:1158 unnatural amino acid incorporation 1125:1158 Our method combines in vitro peptide selection by mRNA display with unnatural amino acid incorporation and chemical attachment of synthetic oligosaccharides. 28935113 1 73 theme useful 80:85 arg1 method 87:92 a useful method 78:92 a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions 78:199 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 1 73 theme useful 80:85 arg1 evolution 65:73 Directed evolution 56:73 Directed evolution 56:73 Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities or functions. 28935113 3 74 theme evolution 375:383 arg1 display 437:443 mammalian-cell display 422:443 mammalian-cell display 422:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 74 theme evolution 375:383 arg1 yeast- 411:416 yeast- 411:416 yeast- 411:416 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 74 theme evolution 375:383 arg1 platforms 385:393 common directed evolution platforms 359:393 common directed evolution platforms such as phage-, yeast-, or mammalian-cell display 359:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 74 theme evolution 375:383 arg1 phage- 403:408 phage- 403:408 phage- 403:408 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 8 75 theme glycopeptide 1311:1322 arg1 libraries 1337:1345 glycopeptide mRNA display libraries 1311:1345 glycopeptide mRNA display libraries containing ~1013 sequences 1311:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 0 76 theme Glycopeptides 22:34 arg1 Evolution 9:17 Directed Evolution 0:17 Directed Evolution of Glycopeptides 0:34 Directed Evolution of Glycopeptides Using mRNA Display. 28935113 2 77 from abundance 217:225 arg1 nature 262:267 nature 262:267 nature 262:267 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 3 78 theme common 359:364 arg1 display 437:443 mammalian-cell display 422:443 mammalian-cell display 422:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 78 theme common 359:364 arg1 yeast- 411:416 yeast- 411:416 yeast- 411:416 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 78 theme common 359:364 arg1 platforms 385:393 common directed evolution platforms 359:393 common directed evolution platforms such as phage-, yeast-, or mammalian-cell display 359:443 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 3 78 theme common 359:364 arg1 phage- 403:408 phage- 403:408 phage- 403:408 However, common directed evolution platforms such as phage-, yeast-, or mammalian-cell display are limited for these applications by several factors. 28935113 8 79 theme display 1329:1335 arg1 libraries 1337:1345 glycopeptide mRNA display libraries 1311:1345 glycopeptide mRNA display libraries containing ~1013 sequences 1311:1372 Here, we provide an updated and optimized protocol for this method, which is designed to create glycopeptide mRNA display libraries containing ~1013 sequences and select them for target binding. 28935113 0 80 theme mRNA 42:45 arg1 Display 47:53 mRNA Display 42:53 mRNA Display 42:53 Directed Evolution of Glycopeptides Using mRNA Display. 28935113 9 81 theme HIV 1556:1558 arg1 gp120 1582:1586 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 2 82 from importance 231:240 arg1 nature 262:267 nature 262:267 nature 262:267 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 5 83 theme homogenous 761:770 arg1 glycoform 772:780 a homogenous glycoform 759:780 a homogenous glycoform 759:780 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 9 84 theme glycoprotein 1569:1580 arg1 gp120 1582:1586 the HIV envelope glycoprotein gp120 1552:1586 the HIV envelope glycoprotein gp120 1552:1586 The target described here is the HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds to cluster of high-mannose oligosaccharides on the HIV envelope glycoprotein gp120; and glycopeptides that mimic this epitope may be useful in HIV vaccine applications. 28935113 4 85 theme glycosylation 525:537 arg1 site 539:542 each glycosylation site 520:542 each glycosylation site 520:542 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 5 86 gly glycosylation 789:801 arg2 sites 803:807 all glycosylation sites 785:807 all glycosylation sites 785:807 Although yeast, mammalian and Escherichia coli cells can be engineered to produce a homogenous glycoform at all glycosylation sites, there are just a few specific glycan structures that can readily be accessed in this manner. 28935113 4 87 theme mammalian 586:594 arg1 cells 596:600 mammalian cells 586:600 mammalian cells 586:600 Glycan structure at each glycosylation site is not genetically encoded, and yeast and mammalian cells produce a heterogeneous mixture of glycoforms at each site on the protein. 28935113 2 88 gly glycoproteins 310:322 arg1 glycoproteins 310:322 glycoproteins 310:322 glycoproteins 310:322 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28935113 6 89 gly glycopeptide 970:981 arg2 glycopeptide 970:981 glycopeptide libraries 970:991 glycopeptide libraries 970:991 Recently, we reported a novel system for the directed evolution of glycopeptide libraries, which could in principle be decorated with any desired glycan. 28935113 2 90 gly glycopeptides 292:304 arg2 glycopeptides 292:304 glycopeptides 292:304 glycopeptides 292:304 Because of the abundance and importance of glycosylation in nature, directed evolution of glycopeptides and glycoproteins is also highly desirable. 28649731 0 0 from Interactions 23:34 arg1 Environment 48:58 Crowding Environment 39:58 Crowding Environment 39:58 Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. 28649731 5 1 theme N-HSQC 651:656 arg1 signals 658:664 the Gal-3 1 H15 N-HSQC signals 635:664 the Gal-3 1 H15 N-HSQC signals 635:664 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 5 2 theme crowders 699:706 arg1 presence 683:690 the presence 679:690 the presence of all crowders 679:706 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 3 3 from event 402:406 arg1 conditions 462:471 crowding conditions 453:471 crowding conditions 453:471 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 7 4 theme signal 1047:1052 arg1 intensity 1054:1062 the signal intensity 1043:1062 the signal intensity of 1 H15 N-HSQC 1043:1078 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 6 5 theme glycosylated 809:820 arg1 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 7 6 theme lactose 1107:1113 arg1 addition 1095:1102 addition 1095:1102 addition of lactose 1095:1113 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 4 7 theme Gal-3 488:492 arg1 interactions 494:505 Gal-3 interactions 488:505 Gal-3 interactions 488:505 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 2 8 theme serum 307:311 arg1 Ficoll 336:341 Ficoll 70 and PEG3350 336:356 Ficoll 336:341 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 8 theme serum 307:311 arg1 PEG3350 350:356 PEG3350 350:356 PEG3350 350:356 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 8 theme serum 307:311 arg1 albumin 313:319 serum albumin 307:319 bovine and human serum albumin (BSA and HSA) 290:333 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 9 9 theme serum 1335:1339 arg1 glycoproteins 1341:1353 serum glycoproteins 1335:1353 serum glycoproteins 1335:1353 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 6 10 theme BSA 844:846 arg1 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 9 11 theme adhesion 1432:1439 arg1 functions 1456:1464 adhesion and signalling functions 1432:1464 adhesion and signalling functions of this protein 1432:1480 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 2 12 theme human 301:305 arg1 Ficoll 336:341 Ficoll 70 and PEG3350 336:356 Ficoll 336:341 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 12 theme human 301:305 arg1 PEG3350 350:356 PEG3350 350:356 PEG3350 350:356 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 12 theme human 301:305 arg1 albumin 313:319 serum albumin 307:319 bovine and human serum albumin (BSA and HSA) 290:333 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 3 13 theme crowding 453:460 arg1 conditions 462:471 crowding conditions 453:471 crowding conditions 453:471 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 9 14 theme cell 1395:1398 arg1 surface 1400:1406 the cell surface 1391:1406 the cell surface 1391:1406 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 6 15 theme PEG3350 969:975 arg1 solutions 956:964 the more viscous solutions 939:964 the more viscous solutions of PEG3350 and Ficoll 70 939:989 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 6 16 located observed 911:918 arg2 that 906:909 that 906:909 that 906:909 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 6 16 located observed 911:918 arg1 presence 927:934 the presence 923:934 the presence of the more viscous solutions of PEG3350 and Ficoll 70 923:989 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 3 17 theme ligand 424:429 arg1 event 402:406 The molecular recognition event 376:406 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions 376:471 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 5 18 theme solution 736:743 arg1 viscosity 745:753 the solution viscosity 732:753 the solution viscosity 732:753 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 1 19 theme Protein-glycan 90:103 arg1 interactions 105:116 Protein-glycan interactions 90:116 Protein-glycan interactions as modulators for quinary structures in crowding environments 90:178 Protein-glycan interactions as modulators for quinary structures in crowding environments were explored. 28649731 0 20 theme Quinary 15:21 arg1 Interactions 23:34 Protein-Glycan Quinary Interactions 0:34 Protein-Glycan Quinary Interactions in Crowding Environment 0:58 Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. 28649731 6 21 theme containing 822:831 arg1 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 9 22 theme signalling 1445:1454 arg1 functions 1456:1464 adhesion and signalling functions 1432:1464 adhesion and signalling functions of this protein 1432:1480 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 8 23 theme binding 1279:1285 arg1 site 1287:1290 the same binding site 1270:1290 the same binding site 1270:1290 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 28649731 0 24 theme Protein-Glycan 0:13 arg1 Interactions 23:34 Protein-Glycan Quinary Interactions 0:34 Protein-Glycan Quinary Interactions in Crowding Environment 0:58 Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. 28649731 4 25 theme chemical 539:546 arg1 CSP 568:570 CSP 568:570 CSP 568:570 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 4 25 theme chemical 539:546 arg1 perturbation 554:565 chemical shift perturbation 539:565 chemical shift perturbation (CSP) 539:571 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 5 26 theme complexes 793:801 arg1 formation 766:774 the formation 762:774 the formation of large protein complexes 762:801 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 8 27 link α2,3-linked 1179:1189 arg1 moieties 1207:1214 their α2,3-linked sialylgalactose moieties 1173:1214 their α2,3-linked sialylgalactose moieties exposed at their surfaces 1173:1240 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 28649731 3 28 theme specific 415:422 arg1 ligand 424:429 the specific ligand 411:429 the specific ligand 411:429 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 3 28 theme specific 415:422 arg1 lactose 432:438 lactose 432:438 lactose 432:438 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 7 29 theme H15 1069:1071 arg1 N-HSQC 1073:1078 1 H15 N-HSQC 1067:1078 1 H15 N-HSQC 1067:1078 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 5 30 theme signals 658:664 arg1 intensity 622:630 The intensity 618:630 The intensity of the Gal-3 1 H15 N-HSQC signals 618:664 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 1 31 from structures 144:153 arg1 environments 167:178 crowding environments 158:178 crowding environments 158:178 Protein-glycan interactions as modulators for quinary structures in crowding environments were explored. 28649731 6 32 gly glycosylated 809:820 arg1 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 4 33 theme N-HSQC 602:607 arg1 signals 609:615 1 H15 N-HSQC signals 596:615 1 H15 N-HSQC signals 596:615 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 7 34 theme N-HSQC 1073:1078 arg1 intensity 1054:1062 the signal intensity 1043:1062 the signal intensity of 1 H15 N-HSQC 1043:1078 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 2 35 theme human 219:223 arg1 Gal-3 237:241 human galectin 3 (Gal-3) and distinct macromolecular crowders 219:279 Gal-3 237:241 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 9 36 theme protein 1474:1480 arg1 functions 1456:1464 adhesion and signalling functions 1432:1464 adhesion and signalling functions of this protein 1432:1480 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 4 37 theme H15 598:600 arg1 signals 609:615 1 H15 N-HSQC signals 596:615 1 H15 N-HSQC signals 596:615 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 4 38 theme shift 548:552 arg1 CSP 568:570 CSP 568:570 CSP 568:570 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 4 38 theme shift 548:552 arg1 perturbation 554:565 chemical shift perturbation 539:565 chemical shift perturbation (CSP) 539:571 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 7 39 gly glycoproteins 1028:1040 arg1 glycoproteins 1028:1040 glycoproteins 1028:1040 glycoproteins 1028:1040 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 2 40 theme macromolecular 257:270 arg1 albumin 313:319 serum albumin 307:319 bovine and human serum albumin (BSA and HSA) 290:333 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 40 theme macromolecular 257:270 arg1 crowders 272:279 human galectin 3 (Gal-3) and distinct macromolecular crowders 219:279 crowders 272:279 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 6 41 theme Ficoll 981:986 arg1 solutions 956:964 the more viscous solutions 939:964 the more viscous solutions of PEG3350 and Ficoll 70 939:989 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 5 42 theme large 779:783 arg1 complexes 793:801 large protein complexes 779:801 large protein complexes 779:801 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 0 43 theme Crowding 39:46 arg1 Environment 48:58 Crowding Environment 39:58 Crowding Environment 39:58 Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. 28649731 8 44 theme α2,3-linked 1179:1189 arg1 moieties 1207:1214 their α2,3-linked sialylgalactose moieties 1173:1214 their α2,3-linked sialylgalactose moieties exposed at their surfaces 1173:1240 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 28649731 2 45 theme distinct 248:255 arg1 albumin 313:319 serum albumin 307:319 bovine and human serum albumin (BSA and HSA) 290:333 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 45 theme distinct 248:255 arg1 crowders 272:279 human galectin 3 (Gal-3) and distinct macromolecular crowders 219:279 crowders 272:279 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 5 46 theme protein 785:791 arg1 complexes 793:801 large protein complexes 779:801 large protein complexes 779:801 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 4 47 theme signals 609:615 arg1 broadening 582:591 line broadening 577:591 line broadening of 1 H15 N-HSQC signals 577:615 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 4 47 theme signals 609:615 arg1 CSP 568:570 CSP 568:570 CSP 568:570 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 4 47 theme signals 609:615 arg1 perturbation 554:565 chemical shift perturbation 539:565 chemical shift perturbation (CSP) 539:571 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 3 48 theme molecular 380:388 arg1 event 402:406 The molecular recognition event 376:406 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions 376:471 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 5 49 theme Gal-3 639:643 arg1 signals 658:664 the Gal-3 1 H15 N-HSQC signals 635:664 the Gal-3 1 H15 N-HSQC signals 635:664 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 8 50 theme same 1274:1277 arg1 site 1287:1290 the same binding site 1270:1290 the same binding site 1270:1290 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 28649731 1 51 theme quinary 136:142 arg1 structures 144:153 quinary structures 136:153 quinary structures in crowding environments 136:178 Protein-glycan interactions as modulators for quinary structures in crowding environments were explored. 28649731 3 52 theme recognition 390:400 arg1 event 402:406 The molecular recognition event 376:406 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions 376:471 The molecular recognition event of the specific ligand, lactose, by Gal-3 in crowding conditions was evaluated. 28649731 6 53 theme signal 867:872 arg1 broadening 874:883 signal broadening 867:883 signal broadening 867:883 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 9 54 theme quinary 1297:1303 arg1 interaction 1305:1315 The quinary interaction 1293:1315 The quinary interaction 1293:1315 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 6 55 theme HSA 852:854 arg1 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 6 56 theme solutions 956:964 arg1 presence 927:934 the presence 923:934 the presence of the more viscous solutions of PEG3350 and Ficoll 70 923:989 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 7 57 contain containing 1017:1026 arg2 glycoproteins 1028:1040 glycoproteins 1028:1040 glycoproteins 1028:1040 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 7 57 contain containing 1017:1026 arg1 samples 1009:1015 the samples 1005:1015 the samples containing glycoproteins 1005:1040 However, for the samples containing glycoproteins, the signal intensity of 1 H15 N-HSQC recovered upon addition of lactose. 28649731 2 58 dep albumin 313:319 arg1 HSA 330:332 HSA 330:332 HSA 330:332 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 58 dep albumin 313:319 arg1 BSA 322:324 BSA 322:324 BSA 322:324 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 5 59 theme H15 647:649 arg1 signals 658:664 the Gal-3 1 H15 N-HSQC signals 635:664 the Gal-3 1 H15 N-HSQC signals 635:664 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 6 60 theme viscous 948:954 arg1 solutions 956:964 the more viscous solutions 939:964 the more viscous solutions of PEG3350 and Ficoll 70 939:989 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 2 61 theme bovine 290:295 arg1 Ficoll 336:341 Ficoll 70 and PEG3350 336:356 Ficoll 336:341 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 61 theme bovine 290:295 arg1 PEG3350 350:356 PEG3350 350:356 PEG3350 350:356 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 2 61 theme bovine 290:295 arg1 albumin 313:319 serum albumin 307:319 bovine and human serum albumin (BSA and HSA) 290:333 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 1 62 theme crowding 158:165 arg1 environments 167:178 crowding environments 158:178 crowding environments 158:178 Protein-glycan interactions as modulators for quinary structures in crowding environments were explored. 28649731 8 63 theme serum 1129:1133 arg1 proteins 1135:1142 serum proteins 1129:1142 serum proteins 1129:1142 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 28649731 4 64 theme line 577:580 arg1 broadening 582:591 line broadening 577:591 line broadening of 1 H15 N-HSQC signals 577:615 Gal-3 interactions were monitored by NMR analysing chemical shift perturbation (CSP) and line broadening of 1 H15 N-HSQC signals. 28649731 2 65 theme galectin 225:232 arg1 Gal-3 237:241 human galectin 3 (Gal-3) and distinct macromolecular crowders 219:279 Gal-3 237:241 The interaction between human galectin 3 (Gal-3) and distinct macromolecular crowders, such as bovine and human serum albumin (BSA and HSA), Ficoll 70 and PEG3350, was scrutinized. 28649731 9 66 gly glycoproteins 1341:1353 arg1 glycoproteins 1341:1353 serum glycoproteins 1335:1353 serum glycoproteins 1335:1353 The quinary interaction between Gal-3 and serum glycoproteins, could help to co-localize Gal-3 at the cell surface, and may play a role in adhesion and signalling functions of this protein. 28649731 6 67 used used 861:864 arg2 samples 833:839 glycosylated containing samples 809:839 glycosylated containing samples of BSA and HSA 809:854 When glycosylated containing samples of BSA and HSA were used, signal broadening was more severe than that observed in the presence of the more viscous solutions of PEG3350 and Ficoll 70. 28649731 0 68 theme NMR 72:74 arg1 Spectroscopy 76:87 NMR Spectroscopy 72:87 NMR Spectroscopy 72:87 Protein-Glycan Quinary Interactions in Crowding Environment Unveiled by NMR Spectroscopy. 28649731 5 69 from increase 720:727 arg1 viscosity 745:753 the solution viscosity 732:753 the solution viscosity 732:753 The intensity of the Gal-3 1 H15 N-HSQC signals decreased in the presence of all crowders, due to the increase in the solution viscosity and to the formation of large protein complexes. 28649731 8 70 theme sialylgalactose 1191:1205 arg1 moieties 1207:1214 their α2,3-linked sialylgalactose moieties 1173:1214 their α2,3-linked sialylgalactose moieties exposed at their surfaces 1173:1240 We show that serum proteins interact with Gal-3, through their α2,3-linked sialylgalactose moieties exposed at their surfaces, competing with lactose for the same binding site. 25936869 6 0 theme siRNA 1241:1245 arg1 transfection 1221:1232 transfection 1221:1232 transfection of BGN siRNA 1221:1245 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 7 1 theme mRNA 1420:1423 arg1 levels 1436:1441 mRNA expression levels 1420:1441 mRNA expression levels 1420:1441 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 4 2 theme size 877:880 arg1 shift 882:886 size shift 877:886 size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation 877:990 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 13 3 theme dermal 2737:2742 arg1 fibroblasts 2744:2754 cultured human dermal fibroblasts 2722:2754 cultured human dermal fibroblasts 2722:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 4 4 theme BGN 840:842 arg1 protein 849:855 BGN core protein 840:855 BGN core protein 840:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 10 5 theme little 1984:1989 arg1 change 1991:1996 a little change 1982:1996 a little change of its molecular weight 1982:2020 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 9 6 theme XYLT2 1739:1743 arg1 down-regulation 1720:1734 down-regulation 1720:1734 down-regulation of XYLT2 1720:1743 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 7 7 theme UV 1390:1391 arg1 irradiation 1393:1403 UV irradiation 1390:1403 UV irradiation 1390:1403 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 9 8 theme siRNA-mediated 1613:1626 arg1 down-regulation 1628:1642 siRNA-mediated down-regulation 1613:1642 siRNA-mediated down-regulation of XYLT1 1613:1651 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 9 theme GAG-attaching 1833:1845 arg1 sites 1854:1858 both GAG-attaching serine sites 1828:1858 both GAG-attaching serine sites of BGN 1828:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 9 theme GAG-attaching 1833:1845 arg1 BGN 1863:1865 BGN 1863:1865 BGN 1863:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 3 10 theme xylosyltransferase 566:583 arg1 XYLT 586:589 xylosyltransferase (XYLT) 1 566:592 xylosyltransferase (XYLT) 1 566:592 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 12 11 theme overexpression 2567:2580 arg1 downregulation 2534:2547 UV-caused downregulation 2524:2547 UV-caused downregulation of both XYLTs and overexpression condition 2524:2590 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 4 12 theme mutants 910:916 arg1 shift 882:886 size shift 877:886 size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation 877:990 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 10 13 theme molecular 2005:2013 arg1 weight 2015:2020 its molecular weight 2001:2020 its molecular weight 2001:2020 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 3 14 theme BGN 739:741 arg1 knockdown 758:766 BGN siRNA-mediated knockdown 739:766 BGN siRNA-mediated knockdown 739:766 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 12 15 theme size 2482:2485 arg1 reduction 2487:2495 size reduction 2482:2495 size reduction 2482:2495 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 5 16 theme BGN 1039:1041 arg1 protein 1043:1049 BGN protein 1039:1049 BGN protein (I-BGN) 1039:1057 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 5 16 theme BGN 1039:1041 arg1 I-BGN 1052:1056 I-BGN 1052:1056 I-BGN 1052:1056 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 3 17 theme irradiation-induced 543:561 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 17 theme irradiation-induced 543:561 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 1 18 theme 42-kDa 207:212 arg1 protein 219:225 a 42-kDa core protein 205:225 a 42-kDa core protein 205:225 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 4 19 theme core 921:924 arg1 size 934:937 core protein size 921:937 core protein size caused by XYLT1 siRNA transfection or UV irradiation 921:990 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 9 20 theme BGN 1863:1865 arg1 sites 1854:1858 both GAG-attaching serine sites 1828:1858 both GAG-attaching serine sites of BGN 1828:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 20 theme BGN 1863:1865 arg1 BGN 1863:1865 BGN 1863:1865 BGN 1863:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 6 21 theme core 1267:1270 arg1 size 1280:1283 the core protein size 1263:1283 the core protein size 1263:1283 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 13 22 theme monoglycosylated 2696:2711 arg1 forms 2713:2717 BGN monoglycosylated forms 2692:2717 BGN monoglycosylated forms 2692:2717 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 10 23 dep sulfate 1933:1939 arg1 proteoglycan 1946:1957 proteoglycan 1946:1957 proteoglycan 1946:1957 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 3 24 theme chondroitinase 772:785 arg1 ABC 787:789 chondroitinase ABC 772:789 chondroitinase ABC (ChABC) 772:797 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 24 theme chondroitinase 772:785 arg1 ChABC 792:796 ChABC 792:796 ChABC 792:796 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 13 25 theme XYLT1 2890:2894 arg1 reduction 2877:2885 UV-mediated stronger reduction 2856:2885 UV-mediated stronger reduction of XYLT1 2856:2894 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 13 26 theme cultured 2722:2729 arg1 fibroblasts 2744:2754 cultured human dermal fibroblasts 2722:2754 cultured human dermal fibroblasts 2722:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 4 27 from activity 813:820 arg1 protein 849:855 BGN core protein 840:855 BGN core protein 840:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 4 28 theme siRNA 955:959 arg1 transfection 961:972 XYLT1 siRNA transfection 949:972 XYLT1 siRNA transfection 949:972 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 12 29 theme incomplete 2604:2613 arg1 glycosylation 2615:2627 incomplete glycosylation 2604:2627 incomplete glycosylation 2604:2627 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 3 30 theme smaller-sized 609:621 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 30 theme smaller-sized 609:621 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 7 31 theme synthesis 1509:1517 arg1 initiation 1485:1494 initiation 1485:1494 initiation of GAG chain synthesis 1485:1517 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 2 32 theme GAG 473:475 arg1 synthesis 483:491 its GAG chain synthesis 469:491 its GAG chain synthesis in cultured human dermal fibroblasts 469:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 0 33 from production 23:32 arg1 fibroblasts 135:145 cultured human dermal fibroblasts 113:145 cultured human dermal fibroblasts 113:145 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 9 34 theme D-BGN 1681:1685 arg1 decrease 1695:1702 the decrease 1691:1702 the decrease of I-BGN 1691:1711 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 34 theme D-BGN 1681:1685 arg1 increase 1669:1676 the increase 1665:1676 the increase of D-BGN 1665:1685 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 7 35 theme GAG 1499:1501 arg1 synthesis 1509:1517 GAG chain synthesis 1499:1517 GAG chain synthesis 1499:1517 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 2 36 theme dermal 511:516 arg1 fibroblasts 518:528 cultured human dermal fibroblasts 496:528 cultured human dermal fibroblasts 496:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 11 37 theme XYLT1 2316:2320 arg1 downregulation 2322:2335 XYLT1 downregulation 2316:2335 XYLT1 downregulation 2316:2335 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 10 38 theme DS 2085:2086 arg1 synthesis 2088:2096 DS synthesis 2085:2096 DS synthesis 2085:2096 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 10 38 theme DS 2085:2086 arg1 factor 2120:2125 a critical factor 2109:2125 a critical factor 2109:2125 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 10 39 theme dermatan 1924:1931 arg1 DS 1942:1943 DS 1942:1943 DS 1942:1943 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 10 39 theme dermatan 1924:1931 arg1 sulfate 1933:1939 Another dermatan sulfate 1916:1939 Another dermatan sulfate (DS) 1916:1944 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 10 39 theme dermatan 1924:1931 arg1 decorin 1960:1966 decorin 1960:1966 decorin 1960:1966 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 0 40 theme cultured 113:120 arg1 fibroblasts 135:145 cultured human dermal fibroblasts 113:145 cultured human dermal fibroblasts 113:145 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 1 41 theme physiological 340:352 arg1 regulations 354:364 many physiological regulations 335:364 many physiological regulations in the skin 335:376 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 2 42 theme cultured 496:503 arg1 fibroblasts 518:528 cultured human dermal fibroblasts 496:528 cultured human dermal fibroblasts 496:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 0 43 theme dermal 128:133 arg1 fibroblasts 135:145 cultured human dermal fibroblasts 113:145 cultured human dermal fibroblasts 113:145 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 5 44 theme core 1063:1066 arg1 form 1076:1079 core protein form 1063:1079 core protein form 1063:1079 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 0 45 theme biglycan 54:61 arg1 production 23:32 UV irradiation-induced production 0:32 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 0:146 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 12 46 dep XYLTs 2557:2561 arg1 condition 2582:2590 condition 2582:2590 condition 2582:2590 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 5 47 theme other 1123:1127 arg1 bands 1143:1147 other smaller-sized bands 1123:1147 other smaller-sized bands 1123:1147 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 13 48 from increase 2680:2687 arg1 fibroblasts 2744:2754 cultured human dermal fibroblasts 2722:2754 cultured human dermal fibroblasts 2722:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 11 49 theme vectors 2239:2245 arg1 XYLT1 2171:2175 XYLT1 2171:2175 XYLT1 2171:2175 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 11 49 theme vectors 2239:2245 arg1 siRNAs 2180:2185 2 siRNAs 2178:2185 2 siRNAs 2178:2185 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 11 49 theme vectors 2239:2245 arg1 forms 2211:2215 wild-type or mutant forms 2191:2215 wild-type or mutant forms of BGN overexpression vectors 2191:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 1 50 from regulations 354:364 arg1 skin 373:376 the skin 369:376 the skin 369:376 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 4 51 theme XYLT1 825:829 arg1 activity 813:820 Differential activity 800:820 Differential activity of XYLT1 and 2 on BGN core protein 800:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 0 52 theme monoglycosylated 37:52 arg1 biglycan 54:61 monoglycosylated biglycan 37:61 monoglycosylated biglycan 37:61 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 5 53 theme cultured 1097:1104 arg1 fibroblasts 1106:1116 cultured fibroblasts 1097:1116 cultured fibroblasts 1097:1116 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 2 54 theme protein 457:463 arg1 changes 442:448 ultraviolet (UV) irradiation-induced changes 405:448 ultraviolet (UV) irradiation-induced changes of BGN protein 405:463 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 2 54 theme protein 457:463 arg1 synthesis 483:491 its GAG chain synthesis 469:491 its GAG chain synthesis in cultured human dermal fibroblasts 469:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 2 55 theme BGN 453:455 arg1 protein 457:463 BGN protein 453:463 BGN protein 453:463 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 3 56 gly monoglycosylated 702:717 arg1 BGN 728:730 BGN 728:730 BGN 728:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 56 gly monoglycosylated 702:717 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 56 gly monoglycosylated 702:717 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 2 57 theme chain 477:481 arg1 synthesis 483:491 its GAG chain synthesis 469:491 its GAG chain synthesis in cultured human dermal fibroblasts 469:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 4 58 theme 2 835:835 arg1 activity 813:820 Differential activity 800:820 Differential activity of XYLT1 and 2 on BGN core protein 800:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 1 59 from functions 321:329 arg1 skin 373:376 the skin 369:376 the skin 369:376 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 1 60 theme structural 295:304 arg1 functions 321:329 structural, space-filling functions 295:329 structural, space-filling functions 295:329 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 13 61 theme XYLT2 2786:2790 arg1 activity 2792:2799 XYLT2 activity 2786:2799 XYLT2 activity 2786:2799 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 11 62 theme core 2413:2416 arg1 protein 2418:2424 BGN core protein 2409:2424 BGN core protein 2409:2424 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 3 63 theme BGN 668:670 arg1 antibodies 672:681 BGN antibodies 668:681 BGN antibodies 668:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 1 64 theme glycosaminoglycan 235:251 arg1 chains 259:264 two glycosaminoglycan (GAG) chains 231:264 two glycosaminoglycan (GAG) chains 231:264 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 7 65 theme expression 1425:1434 arg1 levels 1436:1441 mRNA expression levels 1420:1441 mRNA expression levels 1420:1441 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 13 66 theme UV-mediated 2856:2866 arg1 reduction 2877:2885 UV-mediated stronger reduction 2856:2885 UV-mediated stronger reduction of XYLT1 2856:2894 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 11 67 theme wild-type 2191:2199 arg1 forms 2211:2215 wild-type or mutant forms 2191:2215 wild-type or mutant forms of BGN overexpression vectors 2191:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 8 68 theme XYLT1 1545:1549 arg1 expression 1551:1560 XYLT1 expression 1545:1560 XYLT1 expression 1545:1560 UV-mediated reduction of XYLT1 expression was much stronger than that of XYLT2. 25936869 10 69 theme D-BGN 2143:2147 arg1 formation 2130:2138 formation 2130:2138 formation of D-BGN 2130:2147 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 6 70 theme BGN 1237:1239 arg1 siRNA 1241:1245 BGN siRNA 1237:1245 BGN siRNA 1237:1245 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 10 71 theme XYLT1 2043:2047 arg1 transfection 2055:2066 XYLT1 siRNA transfection 2043:2066 XYLT1 siRNA transfection 2043:2066 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 13 72 theme human 2731:2735 arg1 fibroblasts 2744:2754 cultured human dermal fibroblasts 2722:2754 cultured human dermal fibroblasts 2722:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 10 73 theme critical 2111:2118 arg1 synthesis 2088:2096 DS synthesis 2085:2096 DS synthesis 2085:2096 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 10 73 theme critical 2111:2118 arg1 factor 2120:2125 a critical factor 2109:2125 a critical factor 2109:2125 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 6 74 theme defectively-glycosylated 1335:1358 arg1 they 1326:1329 they 1326:1329 they 1326:1329 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 6 74 theme defectively-glycosylated 1335:1358 arg1 forms 1360:1364 defectively-glycosylated forms 1335:1364 defectively-glycosylated forms of BGN (D-BGN) protein 1335:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 3 75 theme BGN 728:730 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 75 theme BGN 728:730 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 4 76 theme core 844:847 arg1 protein 849:855 BGN core protein 840:855 BGN core protein 840:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 9 77 theme serine 1847:1852 arg1 sites 1854:1858 both GAG-attaching serine sites 1828:1858 both GAG-attaching serine sites of BGN 1828:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 77 theme serine 1847:1852 arg1 BGN 1863:1865 BGN 1863:1865 BGN 1863:1865 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 12 78 theme XYLTs 2557:2561 arg1 downregulation 2534:2547 UV-caused downregulation 2524:2547 UV-caused downregulation of both XYLTs and overexpression condition 2524:2590 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 13 79 theme BGN 2828:2830 arg1 protein 2837:2843 BGN core protein 2828:2843 BGN core protein 2828:2843 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 4 80 theme S47A-BGN 901:908 arg1 mutants 910:916 S42A- and S47A-BGN mutants 891:916 S42A- and S47A-BGN mutants 891:916 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 10 81 theme weight 2015:2020 arg1 change 1991:1996 a little change 1982:1996 a little change of its molecular weight 1982:2020 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 3 82 theme siRNA-mediated 743:756 arg1 knockdown 758:766 BGN siRNA-mediated knockdown 739:766 BGN siRNA-mediated knockdown 739:766 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 6 83 theme protein 1272:1278 arg1 size 1280:1283 the core protein size 1263:1283 the core protein size 1263:1283 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 5 84 theme protein 1043:1049 arg1 form 1031:1034 intact form 1024:1034 intact form of BGN protein (I-BGN) 1024:1057 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 5 84 theme protein 1043:1049 arg1 form 1076:1079 core protein form 1063:1079 core protein form 1063:1079 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 1 85 theme core 214:217 arg1 protein 219:225 a 42-kDa core protein 205:225 a 42-kDa core protein 205:225 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 13 86 theme stronger 2868:2875 arg1 reduction 2877:2885 UV-mediated stronger reduction 2856:2885 UV-mediated stronger reduction of XYLT1 2856:2894 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 12 87 theme UV-caused 2524:2532 arg1 downregulation 2534:2547 UV-caused downregulation 2524:2547 UV-caused downregulation of both XYLTs and overexpression condition 2524:2590 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 2 88 from synthesis 483:491 arg1 fibroblasts 518:528 cultured human dermal fibroblasts 496:528 cultured human dermal fibroblasts 496:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 6 89 theme smaller-sized 1186:1198 arg1 ones 1200:1203 These smaller-sized ones 1180:1203 These smaller-sized ones 1180:1203 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 11 90 theme size 2277:2280 arg1 reduction 2282:2290 size reduction 2277:2290 size reduction 2277:2290 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 9 91 theme I-BGN 1707:1711 arg1 decrease 1695:1702 the decrease 1691:1702 the decrease of I-BGN 1691:1711 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 91 theme I-BGN 1707:1711 arg1 increase 1669:1676 the increase 1665:1676 the increase of D-BGN 1665:1685 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 13 92 theme forms 2713:2717 arg1 increase 2680:2687 UV irradiation-induced increase 2657:2687 UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts 2657:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 13 93 theme activity 2792:2799 arg1 dominance 2773:2781 dominance 2773:2781 dominance 2773:2781 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 10 94 theme UV 2025:2026 arg1 irradiation 2028:2038 UV irradiation 2025:2038 UV irradiation 2025:2038 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 5 95 theme intact 1024:1029 arg1 form 1031:1034 intact form 1024:1034 intact form of BGN protein (I-BGN) 1024:1057 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 3 96 theme protein 623:629 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 96 theme protein 623:629 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 11 97 theme BGN 2220:2222 arg1 vectors 2239:2245 BGN overexpression vectors 2220:2245 BGN overexpression vectors 2220:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 4 98 theme S42A- 891:895 arg1 mutants 910:916 S42A- and S47A-BGN mutants 891:916 S42A- and S47A-BGN mutants 891:916 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 11 99 theme protein 2300:2306 arg1 size 2308:2311 core protein size 2295:2311 core protein size 2295:2311 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 0 100 theme xylosyltransferase 89:106 arg1 downregulation 71:84 downregulation 71:84 downregulation of xylosyltransferase 1 71:108 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 3 101 theme siRNA-mediated 594:607 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 101 theme siRNA-mediated 594:607 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 6 102 with treatment 1288:1296 arg1 ChABC 1303:1307 ChABC 1303:1307 ChABC 1303:1307 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 7 103 theme chain 1503:1507 arg1 synthesis 1509:1517 GAG chain synthesis 1499:1517 GAG chain synthesis 1499:1517 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 4 104 theme XYLT1 949:953 arg1 transfection 961:972 XYLT1 siRNA transfection 949:972 XYLT1 siRNA transfection 949:972 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 10 105 theme siRNA 2049:2053 arg1 transfection 2055:2066 XYLT1 siRNA transfection 2043:2066 XYLT1 siRNA transfection 2043:2066 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 2 106 theme human 505:509 arg1 fibroblasts 518:528 cultured human dermal fibroblasts 496:528 cultured human dermal fibroblasts 496:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 13 107 theme BGN 2692:2694 arg1 forms 2713:2717 BGN monoglycosylated forms 2692:2717 BGN monoglycosylated forms 2692:2717 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 9 108 theme XYLT1 1647:1651 arg1 down-regulation 1628:1642 siRNA-mediated down-regulation 1613:1642 siRNA-mediated down-regulation of XYLT1 1613:1651 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 2 109 from changes 442:448 arg1 fibroblasts 518:528 cultured human dermal fibroblasts 496:528 cultured human dermal fibroblasts 496:528 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 13 110 theme irradiation-induced 2660:2678 arg1 increase 2680:2687 UV irradiation-induced increase 2657:2687 UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts 2657:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 0 111 theme human 122:126 arg1 fibroblasts 135:145 cultured human dermal fibroblasts 113:145 cultured human dermal fibroblasts 113:145 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 1 112 theme many 335:338 arg1 regulations 354:364 many physiological regulations 335:364 many physiological regulations in the skin 335:376 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 11 113 theme mutant 2204:2209 arg1 forms 2211:2215 wild-type or mutant forms 2191:2215 wild-type or mutant forms of BGN overexpression vectors 2191:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 13 114 theme core 2832:2835 arg1 protein 2837:2843 BGN core protein 2828:2843 BGN core protein 2828:2843 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 4 115 theme protein 926:932 arg1 size 934:937 core protein size 921:937 core protein size caused by XYLT1 siRNA transfection or UV irradiation 921:990 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 1 116 dep structural 295:304 arg1 space-filling 307:319 space-filling 307:319 space-filling 307:319 BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. 25936869 9 117 theme them 1910:1913 arg1 them 1910:1913 them 1910:1913 them 1910:1913 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 9 117 theme them 1910:1913 arg1 one 1903:1905 one 1903:1905 one 1903:1905 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 13 118 theme UV 2657:2658 arg1 increase 2680:2687 UV irradiation-induced increase 2657:2687 UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts 2657:2754 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 6 119 gly defectively-glycosylated 1335:1358 arg1 they 1326:1329 they 1326:1329 they 1326:1329 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 6 119 gly defectively-glycosylated 1335:1358 arg1 forms 1360:1364 defectively-glycosylated forms 1335:1364 defectively-glycosylated forms of BGN (D-BGN) protein 1335:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 6 119 gly defectively-glycosylated 1335:1358 arg1 protein 1381:1387 BGN (D-BGN) protein 1369:1387 BGN (D-BGN) protein 1369:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 0 120 theme irradiation-induced 3:21 arg1 production 23:32 UV irradiation-induced production 0:32 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 0:146 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 5 121 theme protein 1068:1074 arg1 form 1076:1079 core protein form 1063:1079 core protein form 1063:1079 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 2 122 theme irradiation-induced 422:440 arg1 changes 442:448 ultraviolet (UV) irradiation-induced changes 405:448 ultraviolet (UV) irradiation-induced changes of BGN protein 405:463 OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. 25936869 10 123 from factor 2120:2125 arg1 formation 2130:2138 formation 2130:2138 formation of D-BGN 2130:2147 Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. 25936869 11 124 theme overexpression 2224:2237 arg1 vectors 2239:2245 BGN overexpression vectors 2220:2245 BGN overexpression vectors 2220:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 6 125 theme D-BGN 1374:1378 arg1 protein 1381:1387 BGN (D-BGN) protein 1369:1387 BGN (D-BGN) protein 1369:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 5 126 theme smaller-sized 1129:1141 arg1 bands 1143:1147 other smaller-sized bands 1123:1147 other smaller-sized bands 1123:1147 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 3 127 theme Western 649:655 arg1 blot 657:660 Western blot 649:660 Western blot using BGN antibodies 649:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 11 128 theme core 2295:2298 arg1 size 2308:2311 core protein size 2295:2311 core protein size 2295:2311 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 6 129 theme BGN 1369:1371 arg1 protein 1381:1387 BGN (D-BGN) protein 1369:1387 BGN (D-BGN) protein 1369:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 4 130 theme UV 977:978 arg1 irradiation 980:990 UV irradiation 977:990 UV irradiation 977:990 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 13 131 gly monoglycosylated 2696:2711 arg1 forms 2713:2717 BGN monoglycosylated forms 2692:2717 BGN monoglycosylated forms 2692:2717 CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1. 25936869 11 132 from S42 2402:2404 arg1 protein 2418:2424 BGN core protein 2409:2424 BGN core protein 2409:2424 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 11 133 with Co-transfection 2150:2164 arg1 XYLT1 2171:2175 XYLT1 2171:2175 XYLT1 2171:2175 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 11 133 with Co-transfection 2150:2164 arg1 siRNAs 2180:2185 2 siRNAs 2178:2185 2 siRNAs 2178:2185 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 11 133 with Co-transfection 2150:2164 arg1 forms 2211:2215 wild-type or mutant forms 2191:2215 wild-type or mutant forms of BGN overexpression vectors 2191:2245 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 5 134 theme UV 1008:1009 arg1 irradiation 1011:1021 UV irradiation 1008:1021 UV irradiation 1008:1021 RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. 25936869 9 135 theme D-BGN 1770:1774 arg1 change 1760:1765 no change 1757:1765 no change of D-BGN and I-BGN 1757:1784 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 8 136 theme UV-mediated 1520:1530 arg1 reduction 1532:1540 UV-mediated reduction 1520:1540 UV-mediated reduction of XYLT1 expression 1520:1560 UV-mediated reduction of XYLT1 expression was much stronger than that of XYLT2. 25936869 7 137 theme 2 1456:1456 arg1 levels 1436:1441 mRNA expression levels 1420:1441 mRNA expression levels 1420:1441 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 0 138 gly monoglycosylated 37:52 arg1 biglycan 54:61 monoglycosylated biglycan 37:61 monoglycosylated biglycan 37:61 UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts. 25936869 9 139 theme I-BGN 1780:1784 arg1 change 1760:1765 no change 1757:1765 no change of D-BGN and I-BGN 1757:1784 Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. 25936869 11 140 theme BGN 2409:2411 arg1 protein 2418:2424 BGN core protein 2409:2424 BGN core protein 2409:2424 Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. 25936869 12 141 theme UV 2432:2433 arg1 irradiation 2435:2445 UV irradiation 2432:2445 UV irradiation 2432:2445 With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. 25936869 7 142 theme XYLT1 1446:1450 arg1 levels 1436:1441 mRNA expression levels 1420:1441 mRNA expression levels 1420:1441 UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. 25936869 8 143 theme expression 1551:1560 arg1 reduction 1532:1540 UV-mediated reduction 1520:1540 UV-mediated reduction of XYLT1 expression 1520:1560 UV-mediated reduction of XYLT1 expression was much stronger than that of XYLT2. 25936869 3 144 theme monoglycosylated 702:717 arg1 forms 719:723 monoglycosylated forms 702:723 monoglycosylated forms of BGN 702:730 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 3 144 theme monoglycosylated 702:717 arg1 bands 631:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands 540:635 UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies 540:681 METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). 25936869 4 145 theme Differential 800:811 arg1 activity 813:820 Differential activity 800:820 Differential activity of XYLT1 and 2 on BGN core protein 800:855 Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. 25936869 6 146 theme protein 1381:1387 arg1 they 1326:1329 they 1326:1329 they 1326:1329 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 25936869 6 146 theme protein 1381:1387 arg1 forms 1360:1364 defectively-glycosylated forms 1335:1364 defectively-glycosylated forms of BGN (D-BGN) protein 1335:1387 These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. 26160412 4 0 theme resulting 763:771 arg1 extracts 773:780 the resulting extracts 759:780 the resulting extracts 759:780 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 11 1 theme major 1828:1832 arg1 proteins 1844:1851 major human CSF proteins 1828:1851 major human CSF proteins 1828:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 7 2 theme site-specific 1179:1191 arg1 information 1193:1203 site-specific information 1179:1203 site-specific information on the N-glycosylation of CSF glycoproteins 1179:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 4 3 from proteins 699:706 arg1 glycopeptides 676:688 Tryptic glycopeptides 668:688 Tryptic glycopeptides from CSF proteins 668:706 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 9 4 theme disorders 1571:1579 arg1 field 1544:1548 the field 1540:1548 the field of neurodegenerative disorders 1540:1579 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 3 5 theme Reproducible 507:518 arg1 profiles 520:527 Reproducible profiles 507:527 Reproducible profiles of CSF N-glycans 507:544 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 7 6 theme versatile 1146:1154 arg1 approach 1156:1163 a versatile approach 1144:1163 a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins 1144:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 8 7 gly N-glycosites 1310:1321 arg2 N-glycosites 1310:1321 55 N-glycosites 1307:1321 55 N-glycosites from 36 glycoproteins 1307:1343 Under these conditions, 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins were tentatively identified. 26160412 2 8 theme cerebrospinal 315:327 arg1 CSF 336:338 CSF 336:338 CSF 336:338 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 8 theme cerebrospinal 315:327 arg1 fluid 329:333 cerebrospinal fluid 315:333 cerebrospinal fluid (CSF) 315:339 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 3 9 theme N-glycans 536:544 arg1 profiles 520:527 Reproducible profiles 507:527 Reproducible profiles of CSF N-glycans 507:544 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 1 10 theme most 198:201 arg1 types 211:215 the most complex types 194:215 the most complex types of post-translational modifications of proteins 194:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 6 11 theme Site-specific 1019:1031 arg1 data 1033:1036 Site-specific data 1019:1036 Site-specific data 1019:1036 Site-specific data were obtained by combining the data from these three experiments. 26160412 4 12 theme Tryptic 668:674 arg1 glycopeptides 676:688 Tryptic glycopeptides 668:688 Tryptic glycopeptides from CSF proteins 668:706 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 0 13 theme combined 16:23 arg1 approach 55:62 a combined glycomics and glycoproteomics approach 14:62 a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids 14:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 1 14 theme types 211:215 arg1 one 187:189 one 187:189 one 187:189 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 1 14 theme types 211:215 arg1 types 211:215 the most complex types 194:215 the most complex types of post-translational modifications of proteins 194:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 8 15 from glycoproteins 1331:1343 arg1 N-glycosites 1310:1321 55 N-glycosites 1307:1321 55 N-glycosites from 36 glycoproteins 1307:1343 Under these conditions, 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins were tentatively identified. 26160412 5 16 theme enriched 972:979 arg1 glycopeptides 981:993 intact enriched glycopeptides 965:993 intact enriched glycopeptides 965:993 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 9 17 theme Special 1374:1380 arg1 emphasis 1382:1389 Special emphasis 1374:1389 Special emphasis 1374:1389 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 3 18 theme time-of-flight 613:626 arg1 spectrometry 633:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 569:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation 569:665 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 2 19 from fluid 329:333 arg1 proteins 301:308 proteins 301:308 proteins from cerebrospinal fluid (CSF) 301:339 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 6 20 from experiments 1091:1101 arg1 data 1069:1072 the data 1065:1072 the data from these three experiments 1065:1101 Site-specific data were obtained by combining the data from these three experiments. 26160412 2 21 theme proteins 488:495 arg1 glycosylation 467:479 the glycosylation 463:479 the glycosylation of CSF proteins 463:495 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 22 theme system 389:394 arg1 disorders 356:364 disorders 356:364 disorders of the central nervous system 356:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 9 23 theme glycoproteins/glycopeptides 1421:1447 arg1 analysis 1409:1416 the analysis 1405:1416 the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans 1405:1478 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 1 24 dep RATIONALE 159:167 arg1 Glycosylation 170:182 Glycosylation 170:182 RATIONALE: Glycosylation 159:182 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 2 25 theme central 373:379 arg1 system 389:394 the central nervous system 369:394 the central nervous system 369:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 26 theme relevant 408:415 arg1 subject 417:423 a highly relevant subject 399:423 a highly relevant subject 399:423 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 26 theme relevant 408:415 arg1 alteration 270:279 The alteration 266:279 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system 266:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 12 27 theme disorders 2021:2029 arg1 understanding 1980:1992 the understanding 1976:1992 the understanding of particular neurological disorders 1976:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 2 28 from alteration 270:279 arg1 relation 344:351 relation 344:351 relation to disorders of the central nervous system 344:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 5 29 theme second 942:947 arg1 one 949:951 the second one 938:951 the second one 938:951 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 7 30 theme CSF 1231:1233 arg1 glycoproteins 1235:1247 CSF glycoproteins 1231:1247 CSF glycoproteins 1231:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 0 31 theme glycomics 25:33 arg1 approach 55:62 a combined glycomics and glycoproteomics approach 14:62 a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids 14:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 10 32 theme glycan 1658:1663 arg1 motifs 1665:1670 such particular glycan motifs 1642:1670 such particular glycan motifs 1642:1670 Using our workflow, only a few proteins were shown to carry such particular glycan motifs. 26160412 11 33 theme proteins 1844:1851 arg1 glycosylation 1811:1823 the site-specific glycosylation 1793:1823 the site-specific glycosylation of major human CSF proteins 1793:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 7 34 gly N-glycosylation 1212:1226 arg1 glycoproteins 1235:1247 CSF glycoproteins 1231:1247 CSF glycoproteins 1231:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 0 35 theme glycoproteomics 39:53 arg1 approach 55:62 a combined glycomics and glycoproteomics approach 14:62 a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids 14:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 9 36 theme potential 1494:1502 arg1 structures 1526:1535 potential biologically relevant structures 1494:1535 potential biologically relevant structures 1494:1535 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 10 37 theme such 1642:1645 arg1 motifs 1665:1670 such particular glycan motifs 1642:1670 such particular glycan motifs 1642:1670 Using our workflow, only a few proteins were shown to carry such particular glycan motifs. 26160412 9 38 theme relevant 1517:1524 arg1 structures 1526:1535 potential biologically relevant structures 1494:1535 potential biologically relevant structures 1494:1535 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 12 39 theme particular 1997:2006 arg1 disorders 2021:2029 particular neurological disorders 1997:2029 particular neurological disorders 1997:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 12 40 theme omics 1938:1942 arg1 approaches 1944:1953 other omics approaches 1932:1953 other omics approaches to delve deeper into the understanding of particular neurological disorders 1932:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 1 41 theme modifications 239:251 arg1 types 211:215 the most complex types 194:215 the most complex types of post-translational modifications of proteins 194:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 5 42 theme first 817:821 arg1 aliquot 823:829 A first aliquot 815:829 A first aliquot 815:829 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 0 43 dep Evaluation 0:9 arg1 Application 123:133 Application 123:133 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 0:157 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 11 44 theme site-specific 1797:1809 arg1 glycosylation 1811:1823 the site-specific glycosylation 1793:1823 the site-specific glycosylation of major human CSF proteins 1793:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 4 45 gly glycopeptides 676:688 arg2 glycopeptides 676:688 Tryptic glycopeptides 668:688 Tryptic glycopeptides from CSF proteins 668:706 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 4 45 gly glycopeptides 676:688 arg1 proteins 699:706 CSF proteins 695:706 CSF proteins 695:706 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 1 46 theme proteins 256:263 arg1 modifications 239:251 post-translational modifications 220:251 post-translational modifications of proteins 220:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 9 47 theme neurodegenerative 1553:1569 arg1 disorders 1571:1579 neurodegenerative disorders 1553:1579 neurodegenerative disorders 1553:1579 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 11 48 theme human 1834:1838 arg1 proteins 1844:1851 major human CSF proteins 1828:1851 major human CSF proteins 1828:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 7 49 theme approach 1156:1163 arg1 development 1129:1139 the development 1125:1139 the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins 1125:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 8 50 gly N-glycopeptides 1278:1292 arg2 N-glycopeptides 1278:1292 124 N-glycopeptides 1274:1292 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins 1274:1343 Under these conditions, 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins were tentatively identified. 26160412 2 51 theme glycans 284:290 arg1 subject 417:423 a highly relevant subject 399:423 a highly relevant subject 399:423 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 51 theme glycans 284:290 arg1 alteration 270:279 The alteration 266:279 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system 266:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 5 52 theme mass 914:917 arg1 spectrometry 919:930 nano-liquid chromatography/tandem mass spectrometry 880:930 nano-liquid chromatography/tandem mass spectrometry 880:930 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 4 53 theme CSF 695:697 arg1 proteins 699:706 CSF proteins 695:706 CSF proteins 695:706 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 3 54 theme CSF 532:534 arg1 N-glycans 536:544 CSF N-glycans 532:544 CSF N-glycans 532:544 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 8 55 theme 124 1274:1276 arg1 N-glycopeptides 1278:1292 124 N-glycopeptides 1274:1292 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins 1274:1343 Under these conditions, 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins were tentatively identified. 26160412 0 56 theme cerebrospinal 138:150 arg1 fluid 152:156 cerebrospinal fluid 138:156 cerebrospinal fluid 138:156 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 8 57 gly glycoproteins 1331:1343 arg1 glycoproteins 1331:1343 36 glycoproteins 1328:1343 36 glycoproteins 1328:1343 Under these conditions, 124 N-glycopeptides representing 55 N-glycosites from 36 glycoproteins were tentatively identified. 26160412 3 58 theme matrix-assisted 569:583 arg1 spectrometry 633:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 569:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation 569:665 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 11 59 gly glycosylation 1811:1823 arg1 proteins 1844:1851 major human CSF proteins 1828:1851 major human CSF proteins 1828:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 5 60 theme nano-liquid 880:890 arg1 spectrometry 919:930 nano-liquid chromatography/tandem mass spectrometry 880:930 nano-liquid chromatography/tandem mass spectrometry 880:930 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 5 61 gly glycopeptides 981:993 arg2 glycopeptides 981:993 intact enriched glycopeptides 965:993 intact enriched glycopeptides 965:993 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 5 62 theme intact 965:970 arg1 glycopeptides 981:993 intact enriched glycopeptides 965:993 intact enriched glycopeptides 965:993 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 0 63 from biofluids 112:120 arg1 present 101:107 present 101:107 present 101:107 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 4 64 theme hydrophilic 730:740 arg1 interaction 742:752 hydrophilic interaction 730:752 hydrophilic interaction 730:752 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 3 65 theme mass 628:631 arg1 spectrometry 633:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 569:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation 569:665 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 12 66 theme final 1858:1862 arg1 objective 1881:1889 The final rather long-term objective 1854:1889 The final rather long-term objective 1854:1889 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 0 67 from present 101:107 arg1 biofluids 112:120 biofluids 112:120 biofluids 112:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 5 68 contain containing 954:963 arg2 glycopeptides 981:993 intact enriched glycopeptides 965:993 intact enriched glycopeptides 965:993 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 5 68 contain containing 954:963 arg1 one 949:951 the second one 938:951 the second one 938:951 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 3 69 theme desorption/ionization 591:611 arg1 spectrometry 633:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 569:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation 569:665 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 10 70 contain carry 1636:1640 arg2 motifs 1665:1670 such particular glycan motifs 1642:1670 such particular glycan motifs 1642:1670 Using our workflow, only a few proteins were shown to carry such particular glycan motifs. 26160412 10 70 contain carry 1636:1640 arg1 proteins 1613:1620 only a few proteins 1602:1620 only a few proteins 1602:1620 Using our workflow, only a few proteins were shown to carry such particular glycan motifs. 26160412 12 71 theme long-term 1871:1879 arg1 objective 1881:1889 The final rather long-term objective 1854:1889 The final rather long-term objective 1854:1889 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 0 72 theme major 81:85 arg1 glycoproteins 87:99 the major glycoproteins 77:99 the major glycoproteins present in biofluids 77:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 9 73 gly glycoproteins/glycopeptides 1421:1447 arg2 glycoproteins/glycopeptides 1421:1447 glycoproteins/glycopeptides 1421:1447 glycoproteins/glycopeptides bearing 'brain-type' N-glycans 1421:1478 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 0 74 theme present 101:107 arg1 glycoproteins 87:99 the major glycoproteins 77:99 the major glycoproteins present in biofluids 77:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 2 75 theme CSF 484:486 arg1 proteins 488:495 CSF proteins 484:495 CSF proteins 484:495 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 4 76 theme equal 799:803 arg1 aliquots 805:812 two equal aliquots 795:812 two equal aliquots 795:812 Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. 26160412 2 77 theme nervous 381:387 arg1 system 389:394 the central nervous system 369:394 the central nervous system 369:394 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 2 78 gly glycosylation 467:479 arg1 proteins 488:495 CSF proteins 484:495 CSF proteins 484:495 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 7 79 theme glycoproteins 1235:1247 arg1 N-glycosylation 1212:1226 the N-glycosylation 1208:1226 the N-glycosylation of CSF glycoproteins 1208:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 9 80 theme brain-type 1458:1467 arg1 N-glycans 1470:1478 'brain-type' N-glycans 1457:1478 'brain-type' N-glycans 1457:1478 Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain-type' N-glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. 26160412 12 81 theme other 1932:1936 arg1 approaches 1944:1953 other omics approaches 1932:1953 other omics approaches to delve deeper into the understanding of particular neurological disorders 1932:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 0 82 gly glycoproteins 87:99 arg1 glycoproteins 87:99 the major glycoproteins 77:99 the major glycoproteins present in biofluids 77:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 1 83 theme complex 203:209 arg1 types 211:215 the most complex types 194:215 the most complex types of post-translational modifications of proteins 194:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 11 84 theme CSF 1840:1842 arg1 proteins 1844:1851 major human CSF proteins 1828:1851 major human CSF proteins 1828:1851 CONCLUSIONS: We developed an approach combining N-glycomics and N-glycoproteomics and underline its usefulness to study the site-specific glycosylation of major human CSF proteins. 26160412 0 85 theme approach 55:62 arg1 Evaluation 0:9 Evaluation 0:9 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 0:157 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 12 86 theme delve 1958:1962 arg1 deeper 1964:1969 delve deeper 1958:1969 delve deeper into the understanding of particular neurological disorders 1958:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 2 87 theme few 435:437 arg1 studies 439:445 only few studies 430:445 only few studies 430:445 The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. 26160412 7 88 from information 1193:1203 arg1 N-glycosylation 1212:1226 the N-glycosylation 1208:1226 the N-glycosylation of CSF glycoproteins 1208:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 26160412 10 89 theme particular 1647:1656 arg1 motifs 1665:1670 such particular glycan motifs 1642:1670 such particular glycan motifs 1642:1670 Using our workflow, only a few proteins were shown to carry such particular glycan motifs. 26160412 5 90 gly deglycosylated 849:862 arg1 aliquot 823:829 A first aliquot 815:829 A first aliquot 815:829 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 12 91 theme neurological 2008:2019 arg1 disorders 2021:2029 particular neurological disorders 1997:2029 particular neurological disorders 1997:2029 The final rather long-term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. 26160412 3 92 theme laser 585:589 arg1 spectrometry 633:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 569:644 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation 569:665 METHODS: Reproducible profiles of CSF N-glycans were first obtained by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after permethylation. 26160412 1 93 theme post-translational 220:237 arg1 modifications 239:251 post-translational modifications 220:251 post-translational modifications of proteins 220:263 RATIONALE: Glycosylation is one of the most complex types of post-translational modifications of proteins. 26160412 5 94 theme chromatography/tandem 892:912 arg1 spectrometry 919:930 nano-liquid chromatography/tandem mass spectrometry 880:930 nano-liquid chromatography/tandem mass spectrometry 880:930 A first aliquot was enzymatically deglycosylated and analyzed by nano-liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. 26160412 0 95 attach present 101:107 arg2 glycoproteins 87:99 the major glycoproteins 77:99 the major glycoproteins present in biofluids 77:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 0 95 attach present 101:107 arg1 biofluids 112:120 biofluids 112:120 biofluids 112:120 Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. 26160412 7 96 gly glycoproteins 1235:1247 arg1 glycoproteins 1235:1247 CSF glycoproteins 1231:1247 CSF glycoproteins 1231:1247 RESULTS: We describe the development of a versatile approach for obtaining site-specific information on the N-glycosylation of CSF glycoproteins. 24632142 5 0 dep study 786:790 arg1 resolve 817:823 resolve 817:823 to resolve this discrepancy 814:840 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 2 1 theme fatal 306:310 arg1 disorder 325:332 a fatal neurological disorder 304:332 a fatal neurological disorder called Canavan disease 304:355 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 5 2 theme human 885:889 arg1 aspartoacylase 891:904 recombinantly-expressed human aspartoacylase 861:904 recombinantly-expressed human aspartoacylase 861:904 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 4 3 theme enzyme 667:672 arg1 structure 649:657 the structure 645:657 the structure of this enzyme 645:672 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 6 4 theme consistent 1090:1099 arg1 picture 1101:1107 a consistent picture 1088:1107 a consistent picture of the structural components of this essential brain enzyme 1088:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 4 5 attach presence 691:698 arg2 components 722:731 any non-amino acid components 703:731 any non-amino acid components 703:731 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 4 5 attach presence 691:698 arg1 site 763:766 the putative glycosylation site 736:766 the putative glycosylation site 736:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 1 6 from brain 151:155 arg1 metabolism 106:115 the metabolism 102:115 the metabolism of an important amino acid in the brain 102:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 5 7 theme recombinantly-expressed 861:883 arg1 aspartoacylase 891:904 recombinantly-expressed human aspartoacylase 861:904 recombinantly-expressed human aspartoacylase 861:904 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 6 8 theme enzyme 1162:1167 arg1 components 1127:1136 the structural components 1112:1136 the structural components of this essential brain enzyme 1112:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 1 9 from acid 139:142 arg1 brain 151:155 the brain 147:155 the brain 147:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 4 10 theme components 722:731 arg1 presence 691:698 the presence 687:698 the presence of any non-amino acid components at the putative glycosylation site 687:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 3 11 theme host 451:454 arg1 expression 456:465 eukaryotic host expression 440:465 eukaryotic host expression 440:465 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 0 12 gly glycoprotein 63:74 arg1 glycoprotein 63:74 a glycoprotein 61:74 a glycoprotein 61:74 Reexamination of aspartoacylase: is this human enzyme really a glycoprotein? 24632142 0 12 gly glycoprotein 63:74 arg1 Reexamination 0:12 Reexamination 0:12 Reexamination of aspartoacylase: 0:31 Reexamination of aspartoacylase: is this human enzyme really a glycoprotein? 24632142 2 13 theme Canavan 341:347 arg1 disease 349:355 Canavan disease 341:355 Canavan disease 341:355 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 3 14 theme differential 413:424 arg1 activity 426:433 differential activity 413:433 differential activity upon eukaryotic host expression 413:465 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 2 15 theme activity 275:282 arg1 symptoms 292:299 the symptoms 288:299 the symptoms of a fatal neurological disorder called Canavan disease 288:355 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 2 15 theme activity 275:282 arg1 loss 267:270 a loss 265:270 a loss of activity 265:282 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 3 16 theme deglycosylation 388:402 arg1 studies 404:410 deglycosylation studies 388:410 deglycosylation studies 388:410 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 5 17 theme extensive 776:784 arg1 study 786:790 A more extensive study 769:790 A more extensive study specifically designed to resolve this discrepancy 769:840 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 6 18 theme brain 1156:1160 arg1 enzyme 1162:1167 this essential brain enzyme 1141:1167 this essential brain enzyme 1141:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 1 19 theme release 167:173 arg1 acetate 175:181 the release acetate 163:181 the release acetate serving as the source for fatty acid biosynthesis 163:231 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 3 20 theme glycan 531:536 arg1 presence 517:524 the presence 513:524 the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase 513:633 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 21 theme eukaryotic 440:449 arg1 expression 456:465 eukaryotic host expression 440:465 eukaryotic host expression 440:465 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 0 22 theme aspartoacylase 17:30 arg1 glycoprotein 63:74 a glycoprotein 61:74 a glycoprotein 61:74 Reexamination of aspartoacylase: is this human enzyme really a glycoprotein? 24632142 0 22 theme aspartoacylase 17:30 arg1 Reexamination 0:12 Reexamination 0:12 Reexamination of aspartoacylase: 0:31 Reexamination of aspartoacylase: is this human enzyme really a glycoprotein? 24632142 4 23 theme putative 740:747 arg1 site 763:766 the putative glycosylation site 736:766 the putative glycosylation site 736:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 3 24 theme aspartoacylase 620:633 arg1 stability 574:582 stability 574:582 stability 574:582 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 24 theme aspartoacylase 620:633 arg1 activity 598:605 catalytic activity 588:605 catalytic activity 588:605 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 4 25 theme non-amino 707:715 arg1 components 722:731 any non-amino acid components 703:731 any non-amino acid components 703:731 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 1 26 theme acid 139:142 arg1 metabolism 106:115 the metabolism 102:115 the metabolism of an important amino acid in the brain 102:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 4 27 gly glycosylation 749:761 arg2 site 763:766 the putative glycosylation site 736:766 the putative glycosylation site 736:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 6 28 theme components 1127:1136 arg1 picture 1101:1107 a consistent picture 1088:1107 a consistent picture of the structural components of this essential brain enzyme 1088:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 5 29 theme bacterial 994:1002 arg1 system 1015:1020 a bacterial expression system 992:1020 a bacterial expression system 992:1020 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 2 30 from Defects 234:240 arg1 enzyme 250:255 this enzyme 245:255 this enzyme 245:255 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 6 31 theme experiments 1064:1074 arg1 interpretations 1035:1049 Alternative interpretations 1023:1049 Alternative interpretations of the prior experiments 1023:1074 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 5 32 gly glycosylated 913:924 arg1 aspartoacylase 891:904 recombinantly-expressed human aspartoacylase 861:904 recombinantly-expressed human aspartoacylase 861:904 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 5 33 theme expression 1004:1013 arg1 system 1015:1020 a bacterial expression system 992:1020 a bacterial expression system 992:1020 A more extensive study specifically designed to resolve this discrepancy has now shown that recombinantly-expressed human aspartoacylase is not glycosylated, but is still fully functional and stable even when produced from a bacterial expression system. 24632142 4 34 theme acid 717:720 arg1 components 722:731 any non-amino acid components 703:731 any non-amino acid components 703:731 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 3 35 theme Extensive 358:366 arg1 evidence 368:375 Extensive evidence 358:375 Extensive evidence 358:375 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 36 theme mammalian 610:618 arg1 aspartoacylase 620:633 mammalian aspartoacylase 610:633 mammalian aspartoacylase 610:633 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 37 theme essential 552:560 arg1 role 562:565 an essential role 549:565 an essential role 549:565 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 38 theme site 471:474 arg1 mutagenesis 485:495 site directed mutagenesis 471:495 site directed mutagenesis 471:495 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 0 39 theme human 41:45 arg1 enzyme 47:52 this human enzyme 36:52 this human enzyme 36:52 Reexamination of aspartoacylase: is this human enzyme really a glycoprotein? 24632142 6 40 theme prior 1058:1062 arg1 experiments 1064:1074 the prior experiments 1054:1074 the prior experiments 1054:1074 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 6 41 theme essential 1146:1154 arg1 enzyme 1162:1167 this essential brain enzyme 1141:1167 this essential brain enzyme 1141:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 3 42 theme directed 476:483 arg1 mutagenesis 485:495 site directed mutagenesis 471:495 site directed mutagenesis 471:495 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 3 43 theme catalytic 588:596 arg1 activity 598:605 catalytic activity 588:605 catalytic activity 588:605 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 1 44 theme important 123:131 arg1 acid 139:142 an important amino acid 120:142 an important amino acid in the brain 120:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 1 45 from metabolism 106:115 arg1 brain 151:155 the brain 147:155 the brain 147:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 4 46 theme glycosylation 749:761 arg1 site 763:766 the putative glycosylation site 736:766 the putative glycosylation site 736:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 1 47 theme amino 133:137 arg1 acid 139:142 an important amino acid 120:142 an important amino acid in the brain 120:155 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 6 48 theme structural 1116:1125 arg1 components 1127:1136 the structural components 1112:1136 the structural components of this essential brain enzyme 1112:1167 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 6 49 theme Alternative 1023:1033 arg1 interpretations 1035:1049 Alternative interpretations 1023:1049 Alternative interpretations of the prior experiments 1023:1074 Alternative interpretations of the prior experiments now present a consistent picture of the structural components of this essential brain enzyme. 24632142 3 50 dep stability 574:582 arg1 the 570:572 the 570:572 the 570:572 Extensive evidence, including deglycosylation studies, differential activity upon eukaryotic host expression and site directed mutagenesis, have supported the presence of a glycan that plays an essential role in the stability and catalytic activity of mammalian aspartoacylase. 24632142 2 51 theme disorder 325:332 arg1 symptoms 292:299 the symptoms 288:299 the symptoms of a fatal neurological disorder called Canavan disease 288:355 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 2 51 theme disorder 325:332 arg1 loss 267:270 a loss 265:270 a loss of activity 265:282 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 1 52 theme fatty 209:213 arg1 biosynthesis 220:231 fatty acid biosynthesis 209:231 fatty acid biosynthesis 209:231 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 24632142 4 53 from site 763:766 arg1 presence 691:698 the presence 687:698 the presence of any non-amino acid components at the putative glycosylation site 687:766 However, the structure of this enzyme did not show the presence of any non-amino acid components at the putative glycosylation site. 24632142 2 54 theme neurological 312:323 arg1 disorder 325:332 a fatal neurological disorder 304:332 a fatal neurological disorder called Canavan disease 304:355 Defects in this enzyme lead to a loss of activity and the symptoms of a fatal neurological disorder called Canavan disease. 24632142 1 55 theme acid 215:218 arg1 biosynthesis 220:231 fatty acid biosynthesis 209:231 fatty acid biosynthesis 209:231 Aspartoacylase catalyzes the metabolism of an important amino acid in the brain, with the release acetate serving as the source for fatty acid biosynthesis. 26590834 0 0 theme terminal 69:76 arg1 sugar 78:82 the terminal sugar 65:82 the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis 65:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 2 1 theme cluster 549:555 arg1 mmp1089-1094 557:568 a six-gene cluster mmp1089-1094 538:568 a six-gene cluster mmp1089-1094 538:568 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 2 2 theme N-glycosylation 670:684 arg1 pathway 686:692 the archaellin N-glycosylation pathway 655:692 the archaellin N-glycosylation pathway 655:692 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 1 3 theme multiple 255:262 arg1 sites 264:268 multiple sites 255:268 multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species 255:520 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 2 4 theme six-gene 540:547 arg1 mmp1089-1094 557:568 a six-gene cluster mmp1089-1094 538:568 a six-gene cluster mmp1089-1094 538:568 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 2 5 theme archaellin 659:668 arg1 pathway 686:692 the archaellin N-glycosylation pathway 655:692 the archaellin N-glycosylation pathway 655:692 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 5 6 theme sugar 1195:1199 arg1 residue 1201:1207 the terminal sugar residue 1182:1207 the terminal sugar residue 1182:1207 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 6 theme sugar 1195:1199 arg1 missing 1266:1272 missing 1266:1272 missing 1266:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 8 7 theme N-glycosylation 1837:1851 arg1 pathway 1853:1859 the archaellin N-glycosylation pathway 1822:1859 the archaellin N-glycosylation pathway 1822:1859 Based on its demonstrated involvement in the archaellin N-glycosylation pathway, we designated mmp1090 as aglW. 26590834 6 8 theme terminal 1402:1409 arg1 sugar 1411:1415 the unique terminal sugar 1391:1415 the unique terminal sugar of the archaellin N-glycan 1391:1442 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 5 9 theme third 1241:1245 arg1 residue 1253:1259 the third sugar residue 1237:1259 the third sugar residue 1237:1259 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 2 10 theme potential 716:724 arg1 involvement 726:736 its potential involvement 712:736 its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway 712:800 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 5 11 theme spectrometry 1061:1072 arg1 analysis 1074:1081 mass spectrometry analysis 1056:1081 mass spectrometry analysis of purified archaella 1056:1103 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 8 12 theme archaellin 1826:1835 arg1 pathway 1853:1859 the archaellin N-glycosylation pathway 1822:1859 the archaellin N-glycosylation pathway 1822:1859 Based on its demonstrated involvement in the archaellin N-glycosylation pathway, we designated mmp1090 as aglW. 26590834 0 13 theme sugar 78:82 arg1 pathway 54:60 the biosynthesis pathway 37:60 the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis 37:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 1 14 theme unique 392:397 arg1 -2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 409:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 14 theme unique 392:397 arg1 Sug 383:385 Sug 383:385 Sug 383:385 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 6 15 theme gene 1280:1283 arg1 annotation 1285:1294 gene annotation 1280:1294 gene annotation 1280:1294 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 0 16 from maripaludis 144:154 arg1 pathway 54:60 the biosynthesis pathway 37:60 the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis 37:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 0 17 theme archaellin 91:100 arg1 tetrasaccharide 111:125 the archaellin N-linked tetrasaccharide 87:125 the archaellin N-linked tetrasaccharide 87:125 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 1 18 theme sugar 399:403 arg1 -2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 409:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 18 theme sugar 399:403 arg1 Sug 383:385 Sug 383:385 Sug 383:385 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 6 19 theme UDP-glucose 1350:1360 arg1 MMP1090 1337:1343 MMP1090 1337:1343 MMP1090 1337:1343 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 6 19 theme UDP-glucose 1350:1360 arg1 4-epimerase 1362:1372 a UDP-glucose 4-epimerase 1348:1372 a UDP-glucose 4-epimerase 1348:1372 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 8 20 from involvement 1807:1817 arg1 pathway 1853:1859 the archaellin N-glycosylation pathway 1822:1859 the archaellin N-glycosylation pathway 1822:1859 Based on its demonstrated involvement in the archaellin N-glycosylation pathway, we designated mmp1090 as aglW. 26590834 5 21 theme purified 1086:1093 arg1 archaella 1095:1103 purified archaella 1086:1103 purified archaella 1086:1103 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 7 22 located observed 1650:1657 arg1 contrast 1576:1583 contrast 1576:1583 contrast 1576:1583 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 7 22 located observed 1650:1657 arg2 phenotype 1600:1608 no detectable phenotype 1586:1608 no detectable phenotype related to archaellin glycosylation 1586:1644 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 7 22 located observed 1650:1657 arg1 mutants 1662:1668 mutants 1662:1668 mutants deleted for either mmp1091 or mmp1092 1662:1706 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 1 23 theme 5S 406:407 arg1 -2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 409:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 23 theme 5S 406:407 arg1 Sug 383:385 Sug 383:385 Sug 383:385 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 2 24 theme archaellin 745:754 arg1 N-glycosylation 756:770 the archaellin N-glycosylation or sugar biosynthesis pathway 741:800 N-glycosylation 756:770 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 5 25 theme electron 1032:1039 arg1 microscopy 1041:1050 electron microscopy 1032:1050 electron microscopy 1032:1050 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 0 26 link N-linked 102:109 arg1 tetrasaccharide 111:125 the archaellin N-linked tetrasaccharide 87:125 the archaellin N-linked tetrasaccharide 87:125 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 1 27 theme N-linked 278:285 arg1 tetrasaccharide 287:301 an N-linked tetrasaccharide 275:301 an N-linked tetrasaccharide 275:301 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 3 28 theme genes 837:841 arg1 co-transcription 807:822 The co-transcription 803:822 The co-transcription of these six genes 803:841 The co-transcription of these six genes was demonstrated by RT-PCR. 26590834 7 29 theme archaellin 1621:1630 arg1 glycosylation 1632:1644 archaellin glycosylation 1621:1644 archaellin glycosylation 1621:1644 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 5 30 from missing 1266:1272 arg1 N-glycan 1164:1171 a truncated N-glycan 1152:1171 a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing 1152:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 0 31 theme gene 20:23 arg1 Identification 0:13 Identification 0:13 Identification of a gene 0:23 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 6 32 theme bioinformatic 1300:1312 arg1 analyses 1314:1321 bioinformatic analyses 1300:1321 bioinformatic analyses 1300:1321 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 0 33 theme tetrasaccharide 111:125 arg1 sugar 78:82 the terminal sugar 65:82 the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis 65:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 1 34 with tetrasaccharide 287:301 arg1 structure 312:320 the structure 308:320 the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc 308:374 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 35 located found 488:492 arg2 Sug 383:385 Sug 383:385 Sug 383:385 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 35 located found 488:492 arg1 species 514:520 this species 509:520 this species 509:520 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 1 35 located found 488:492 arg2 -2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 409:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose 390:478 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 0 36 theme N-linked 102:109 arg1 tetrasaccharide 111:125 the archaellin N-linked tetrasaccharide 87:125 the archaellin N-linked tetrasaccharide 87:125 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 7 37 dep attempts 1714:1721 arg1 unsuccessful 1767:1778 unsuccessful 1767:1778 were unsuccessful 1762:1778 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 5 38 theme mass 1056:1059 arg1 analysis 1074:1081 mass spectrometry analysis 1056:1081 mass spectrometry analysis of purified archaella 1056:1103 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 2 39 theme regions 602:608 arg1 one 583:585 one 583:585 one 583:585 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 2 39 theme regions 602:608 arg1 regions 602:608 the genomic regions 590:608 the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway 590:692 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 7 40 theme detectable 1589:1598 arg1 phenotype 1600:1608 no detectable phenotype 1586:1608 no detectable phenotype related to archaellin glycosylation 1586:1644 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 5 41 theme sugar 1247:1251 arg1 residue 1253:1259 the third sugar residue 1237:1259 the third sugar residue 1237:1259 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 2 42 theme genomic 594:600 arg1 regions 602:608 the genomic regions 590:608 the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway 590:692 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 5 43 theme terminal 1186:1193 arg1 residue 1201:1207 the terminal sugar residue 1182:1207 the terminal sugar residue 1182:1207 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 43 theme terminal 1186:1193 arg1 missing 1266:1272 missing 1266:1272 missing 1266:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 44 mod modified 1138:1145 arg3 N-glycan 1164:1171 a truncated N-glycan 1152:1171 a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing 1152:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 44 mod modified 1138:1145 arg1 archaellins 1121:1131 the archaellins 1117:1131 the archaellins 1117:1131 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 0 45 theme biosynthesis 41:52 arg1 pathway 54:60 the biosynthesis pathway 37:60 the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis 37:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 6 46 theme archaellin 1424:1433 arg1 N-glycan 1435:1442 the archaellin N-glycan 1420:1442 the archaellin N-glycan 1420:1442 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 0 47 from pathway 54:60 arg1 maripaludis 144:154 Methanococcus maripaludis 130:154 Methanococcus maripaludis 130:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 2 48 theme biosynthesis 781:792 arg1 pathway 794:800 the archaellin N-glycosylation or sugar biosynthesis pathway 741:800 pathway 794:800 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 6 49 theme N-glycan 1435:1442 arg1 sugar 1411:1415 the unique terminal sugar 1391:1415 the unique terminal sugar of the archaellin N-glycan 1391:1442 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 1 50 theme Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc 325:374 arg1 structure 312:320 the structure 308:320 the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc 308:374 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 6 51 from step 1533:1536 arg1 synthesis 1541:1549 synthesis 1541:1549 synthesis catalysed by MMP1090 1541:1570 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 2 52 theme sugar 775:779 arg1 pathway 794:800 the archaellin N-glycosylation or sugar biosynthesis pathway 741:800 pathway 794:800 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 5 53 attach linked 1227:1232 arg2 missing 1266:1272 missing 1266:1272 missing 1266:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 53 attach linked 1227:1232 arg2 threonine 1217:1225 the threonine 1213:1225 the threonine linked to the third sugar residue 1213:1259 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 53 attach linked 1227:1232 arg1 residue 1253:1259 the third sugar residue 1237:1259 the third sugar residue 1237:1259 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 7 54 theme related 1610:1616 arg1 phenotype 1600:1608 no detectable phenotype 1586:1608 no detectable phenotype related to archaellin glycosylation 1586:1644 In contrast, no detectable phenotype related to archaellin glycosylation was observed in mutants deleted for either mmp1091 or mmp1092 while attempts to delete mmp1089, mmp1093 and mmp1094 were unsuccessful. 26590834 6 55 theme unique 1395:1400 arg1 sugar 1411:1415 the unique terminal sugar 1391:1415 the unique terminal sugar of the archaellin N-glycan 1391:1442 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 5 56 theme truncated 1154:1162 arg1 N-glycan 1164:1171 a truncated N-glycan 1152:1171 a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing 1152:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 57 theme archaella 1095:1103 arg1 analysis 1074:1081 mass spectrometry analysis 1056:1081 mass spectrometry analysis of purified archaella 1056:1103 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 1 58 with sites 264:268 arg1 tetrasaccharide 287:301 an N-linked tetrasaccharide 275:301 an N-linked tetrasaccharide 275:301 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 2 59 from involvement 726:736 arg1 pathway 794:800 the archaellin N-glycosylation or sugar biosynthesis pathway 741:800 pathway 794:800 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 2 59 from involvement 726:736 arg1 N-glycosylation 756:770 the archaellin N-glycosylation or sugar biosynthesis pathway 741:800 N-glycosylation 756:770 In this study, a six-gene cluster mmp1089-1094, neighboring one of the genomic regions already known to contain genes involved with the archaellin N-glycosylation pathway, was examined for its potential involvement in the archaellin N-glycosylation or sugar biosynthesis pathway. 26590834 6 60 theme early 1527:1531 arg1 step 1533:1536 an essential early step 1514:1536 an essential early step in synthesis catalysed by MMP1090 1514:1570 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 5 61 from N-glycan 1164:1171 arg1 residue 1201:1207 the terminal sugar residue 1182:1207 the terminal sugar residue 1182:1207 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 61 from N-glycan 1164:1171 arg1 missing 1266:1272 missing 1266:1272 missing 1266:1272 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 5 61 from N-glycan 1164:1171 arg1 threonine 1217:1225 the threonine 1213:1225 the threonine linked to the third sugar residue 1213:1259 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 0 62 from sugar 78:82 arg1 maripaludis 144:154 Methanococcus maripaludis 130:154 Methanococcus maripaludis 130:154 Identification of a gene involved in the biosynthesis pathway of the terminal sugar of the archaellin N-linked tetrasaccharide in Methanococcus maripaludis. 26590834 4 63 theme in-frame 891:898 arg1 deletion 900:907 an in-frame deletion 888:907 an in-frame deletion 888:907 Mutants carrying an in-frame deletion in mmp1090, mmp1091 or mmp1092 were successfully generated. 26590834 5 64 theme Δmmp1090 973:980 arg1 mutant 991:996 The Δmmp1090 deletion mutant 969:996 The Δmmp1090 deletion mutant 969:996 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 8 65 theme demonstrated 1794:1805 arg1 involvement 1807:1817 its demonstrated involvement 1790:1817 its demonstrated involvement in the archaellin N-glycosylation pathway 1790:1859 Based on its demonstrated involvement in the archaellin N-glycosylation pathway, we designated mmp1090 as aglW. 26590834 6 66 theme essential 1517:1525 arg1 step 1533:1536 an essential early step 1514:1536 an essential early step in synthesis catalysed by MMP1090 1514:1570 Both gene annotation and bioinformatic analyses indicate that MMP1090 is a UDP-glucose 4-epimerase, suggesting that the unique terminal sugar of the archaellin N-glycan might be synthesised from UDP-glucose or UDP-N-acetylglucosamine with an essential early step in synthesis catalysed by MMP1090. 26590834 1 67 link N-linked 278:285 arg1 tetrasaccharide 287:301 an N-linked tetrasaccharide 275:301 an N-linked tetrasaccharide 275:301 In Methanococcus maripaludis, the three archaellins which comprise the archaellum are modified at multiple sites with an N-linked tetrasaccharide with the structure of Sug-4-β-ManNAc3NAmA6Thr-4-β-GlcNAc3NAcA-3-β-GalNAc, where Sug is a unique sugar (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-L-erythro-hexos-5-ulo-1,5-pyranose, so far found exclusively in this species. 26590834 5 68 theme deletion 982:989 arg1 mutant 991:996 The Δmmp1090 deletion mutant 969:996 The Δmmp1090 deletion mutant 969:996 The Δmmp1090 deletion mutant was archaellated when examined by electron microscopy and mass spectrometry analysis of purified archaella showed that the archaellins were modified with a truncated N-glycan in which the terminal sugar residue and the threonine linked to the third sugar residue were missing. 26590834 4 69 contain carrying 879:886 arg1 Mutants 871:877 Mutants 871:877 Mutants carrying an in-frame deletion in mmp1090, mmp1091 or mmp1092 871:938 Mutants carrying an in-frame deletion in mmp1090, mmp1091 or mmp1092 were successfully generated. 26590834 4 69 contain carrying 879:886 arg2 deletion 900:907 an in-frame deletion 888:907 an in-frame deletion 888:907 Mutants carrying an in-frame deletion in mmp1090, mmp1091 or mmp1092 were successfully generated. 24806200 2 0 dep portion 533:539 arg1 presence 498:505 presence 498:505 presence 498:505 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 2 0 dep portion 533:539 arg1 the 494:496 the 494:496 the 494:496 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 2 1 theme spaced 568:573 arg1 glycans 584:590 four closely spaced O-linked glycans 555:590 four closely spaced O-linked glycans 555:590 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 2 2 link O-linked 575:582 arg1 glycans 584:590 four closely spaced O-linked glycans 555:590 four closely spaced O-linked glycans 555:590 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 8 3 theme individual 1620:1629 arg1 fragments 1644:1652 the individual glycopeptide fragments 1616:1652 the individual glycopeptide fragments 1616:1652 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 7 4 theme spaced 1462:1467 arg1 sites 1485:1489 closely spaced O-glycosylation sites 1454:1489 closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1454:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 8 5 theme fragments 1644:1652 arg1 assembly 1604:1611 The final assembly 1594:1611 The final assembly of the individual glycopeptide fragments 1594:1652 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 3 6 theme infertility 694:704 arg1 treatment 681:689 the treatment 677:689 the treatment of infertility 677:704 These glycoproteins play important roles in reproduction and are used clinically in the treatment of infertility. 24806200 7 7 theme installation 1389:1400 arg1 fragment 1352:1359 the N-terminal fragment 1337:1359 the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1337:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 6 8 theme native 1217:1222 arg1 sites 1238:1242 all native glycosylation sites 1213:1242 all native glycosylation sites 1213:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 7 9 theme acid 1434:1437 arg1 cassettes 1439:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 4 10 theme significant 797:807 arg1 interest 809:816 significant interest 797:816 significant interest 797:816 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 7 11 dep access 1547:1552 arg1 portion 1569:1575 the C-terminal portion 1554:1575 the C-terminal portion of the molecule 1554:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 8 12 theme complex 1740:1746 arg1 β-hCG 1776:1780 β-hCG 1776:1780 β-hCG 1776:1780 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 12 theme complex 1740:1746 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 12 theme complex 1740:1746 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 12 theme complex 1740:1746 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 1 13 theme human 155:159 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 13 theme human 155:159 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 13 theme human 155:159 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 13 theme human 155:159 arg1 hCG 185:187 hCG 185:187 hCG 185:187 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 7 14 from steps 1249:1253 arg1 syntheses 1264:1272 these syntheses 1258:1272 these syntheses 1258:1272 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 15 theme β-hCG 1364:1368 arg1 fragment 1352:1359 the N-terminal fragment 1337:1359 the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1337:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 5 16 theme glycoproteins 1025:1037 arg1 samples 1008:1014 homogeneous samples 996:1014 homogeneous samples of these glycoproteins 996:1037 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 8 17 theme glycoprotein 1754:1765 arg1 β-hCG 1776:1780 β-hCG 1776:1780 β-hCG 1776:1780 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 17 theme glycoprotein 1754:1765 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 17 theme glycoprotein 1754:1765 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 17 theme glycoprotein 1754:1765 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 5 18 theme clinical 861:868 arg1 applications 870:881 clinical applications 861:881 clinical applications 861:881 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 4 19 theme cancer 841:846 arg1 vaccines 848:855 cancer vaccines 841:855 cancer vaccines 841:855 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 5 20 theme homogeneous 996:1006 arg1 samples 1008:1014 homogeneous samples 996:1014 homogeneous samples of these glycoproteins 996:1037 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 6 21 gly glycosylation 1224:1236 arg2 sites 1238:1242 all native glycosylation sites 1213:1242 all native glycosylation sites 1213:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 2 22 theme extended 513:520 arg1 portion 533:539 an extended C-terminal portion 510:539 an extended C-terminal portion incorporating four closely spaced O-linked glycans 510:590 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 7 23 theme glycosylation 1311:1323 arg1 route 1328:1332 a successful double Lansbury glycosylation en route 1282:1332 a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1282:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 1 24 theme unique 284:289 arg1 β-subunit 291:299 a unique β-subunit 282:299 a unique β-subunit 282:299 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 24 theme unique 284:289 arg1 subunits 245:252 two subunits 241:252 two subunits 241:252 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 25 theme luteinizing 125:135 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 25 theme luteinizing 125:135 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 25 theme luteinizing 125:135 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 8 26 gly glycoprotein 1754:1765 arg1 glycoprotein 1754:1765 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 27 theme related 1806:1812 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 27 theme related 1806:1812 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 27 theme related 1806:1812 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 27 theme related 1806:1812 arg1 β-hLH 1825:1829 β-hLH 1825:1829 β-hLH 1825:1829 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 0 28 theme chorionic 67:75 arg1 gonadotropin 77:88 chorionic gonadotropin 67:88 chorionic gonadotropin (hCG) 67:94 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 0 28 theme chorionic 67:75 arg1 hCG 91:93 hCG 91:93 hCG 91:93 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 6 29 theme convergent 1136:1145 arg1 syntheses 1147:1155 the highly convergent syntheses 1125:1155 the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites 1125:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 1 30 gly glycoprotein 200:211 arg1 glycoprotein 200:211 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 8 31 theme glycopeptide 1631:1642 arg1 fragments 1644:1652 the individual glycopeptide fragments 1616:1652 the individual glycopeptide fragments 1616:1652 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 5 32 theme glycoforms 931:940 arg1 mixtures 919:926 mixtures 919:926 mixtures of glycoforms due to limitations of biological methods 919:981 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 5 32 theme glycoforms 931:940 arg1 hormones 890:897 these hormones 884:897 these hormones 884:897 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 7 33 theme Lansbury 1302:1309 arg1 route 1328:1332 a successful double Lansbury glycosylation en route 1282:1332 a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1282:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 1 34 dep hormone 137:143 arg1 hLH 146:148 hLH 146:148 hLH 146:148 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 0 35 theme Chemical 0:7 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 0:117 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 8 36 theme discrete 1835:1842 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 36 theme discrete 1835:1842 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 36 theme discrete 1835:1842 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 7 37 theme successful 1284:1293 arg1 route 1328:1332 a successful double Lansbury glycosylation en route 1282:1332 a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1282:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 2 38 theme similarity 391:400 arg1 degree 372:377 a high degree 365:377 a high degree of sequence similarity 365:400 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 6 39 theme homogeneous 1160:1170 arg1 β-hLH 1172:1176 homogeneous β-hLH 1160:1176 homogeneous β-hLH 1160:1176 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 0 40 dep luteinizing 45:55 arg1 hormones 109:116 glycoprotein hormones 96:116 glycoprotein hormones 96:116 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 0 40 dep luteinizing 45:55 arg1 hLH 58:60 hLH 58:60 hLH 58:60 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 2 41 theme common 426:431 arg1 site 449:452 a common N-glycosylation site 424:452 a common N-glycosylation site 424:452 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 2 42 theme high 367:370 arg1 degree 372:377 a high degree 365:377 a high degree of sequence similarity 365:400 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 0 43 theme β-subunit 26:34 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 0:117 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 1 44 theme identical 258:266 arg1 α-subunit 268:276 an identical α-subunit 255:276 an identical α-subunit 255:276 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 44 theme identical 258:266 arg1 subunits 245:252 two subunits 241:252 two subunits 241:252 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 5 45 theme methods 975:981 arg1 limitations 949:959 limitations 949:959 limitations of biological methods 949:981 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 4 46 theme cancers 770:776 arg1 variety 759:765 a variety 757:765 a variety of cancers 757:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 4 46 theme cancers 770:776 arg1 cancers 770:776 cancers 770:776 cancers 770:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 7 47 theme single 1496:1501 arg1 synthesis 1534:1542 a single, high-yielding solid-supported synthesis 1494:1542 a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1494:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 4 48 theme tumor 741:745 arg1 marker 747:752 a tumor marker 739:752 a tumor marker in a variety of cancers 739:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 6 49 theme synthesis 1077:1085 arg1 tools 1059:1063 the powerful tools 1046:1063 the powerful tools of chemical synthesis 1046:1085 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 3 50 used used 658:661 arg2 glycoproteins 599:611 These glycoproteins 593:611 These glycoproteins 593:611 These glycoproteins play important roles in reproduction and are used clinically in the treatment of infertility. 24806200 8 51 theme stepwise 1665:1672 arg1 strategy 1699:1706 a stepwise native chemical ligation strategy 1663:1706 a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms 1663:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 1 52 theme human 194:198 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 52 theme human 194:198 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 52 theme human 194:198 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 7 53 theme O-glycosylation 1469:1483 arg1 sites 1485:1489 closely spaced O-glycosylation sites 1454:1489 closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1454:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 8 54 theme chemical 1681:1688 arg1 strategy 1699:1706 a stepwise native chemical ligation strategy 1663:1706 a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms 1663:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 6 55 theme powerful 1050:1057 arg1 tools 1059:1063 the powerful tools 1046:1063 the powerful tools of chemical synthesis 1046:1085 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 7 56 dep single 1496:1501 arg1 high-yielding 1504:1516 high-yielding 1504:1516 high-yielding 1504:1516 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 57 theme cassettes 1439:1447 arg1 β-hCG 1364:1368 β-hCG 1364:1368 β-hCG 1364:1368 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 57 theme cassettes 1439:1447 arg1 installation 1389:1400 the sequential installation 1374:1400 the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1374:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 58 theme O-linked 1410:1417 arg1 cassettes 1439:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 3 59 gly glycoproteins 599:611 arg1 glycoproteins 599:611 These glycoproteins 593:611 These glycoproteins 593:611 These glycoproteins play important roles in reproduction and are used clinically in the treatment of infertility. 24806200 6 60 theme glycosylation 1224:1236 arg1 sites 1238:1242 all native glycosylation sites 1213:1242 all native glycosylation sites 1213:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 7 61 theme glycosyl-amino 1419:1432 arg1 cassettes 1439:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 0 62 gly glycoprotein 96:107 arg1 glycoprotein 96:107 glycoprotein hormones 96:116 glycoprotein hormones 96:116 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 8 63 theme longest 1723:1729 arg1 β-hCG 1776:1780 β-hCG 1776:1780 β-hCG 1776:1780 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 63 theme longest 1723:1729 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 63 theme longest 1723:1729 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 63 theme longest 1723:1729 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 2 64 theme O-linked 575:582 arg1 glycans 584:590 four closely spaced O-linked glycans 555:590 four closely spaced O-linked glycans 555:590 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 7 65 theme sequential 1378:1387 arg1 installation 1389:1400 the sequential installation 1374:1400 the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1374:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 0 66 theme glycoprotein 96:107 arg1 hormones 109:116 glycoprotein hormones 96:116 glycoprotein hormones 96:116 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 1 67 theme chorionic 161:169 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 67 theme chorionic 161:169 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 67 theme chorionic 161:169 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 67 theme chorionic 161:169 arg1 hCG 185:187 hCG 185:187 hCG 185:187 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 8 68 theme human 1748:1752 arg1 β-hCG 1776:1780 β-hCG 1776:1780 β-hCG 1776:1780 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 68 theme human 1748:1752 arg1 homologue 1814:1822 its closely related homologue 1794:1822 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 68 theme human 1748:1752 arg1 hormone 1767:1773 human glycoprotein hormone 1748:1773 the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) 1719:1830 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 68 theme human 1748:1752 arg1 glycoforms 1844:1853 discrete glycoforms 1835:1853 discrete glycoforms 1835:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 6 69 theme model 1196:1200 arg1 glycans 1202:1208 model glycans 1196:1208 model glycans 1196:1208 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 7 70 from sites 1485:1489 arg1 synthesis 1534:1542 a single, high-yielding solid-supported synthesis 1494:1542 a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1494:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 2 71 theme C-terminal 522:531 arg1 portion 533:539 an extended C-terminal portion 510:539 an extended C-terminal portion incorporating four closely spaced O-linked glycans 510:590 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 7 72 theme N-terminal 1341:1350 arg1 fragment 1352:1359 the N-terminal fragment 1337:1359 the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1337:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 6 73 theme β-hCG 1182:1186 arg1 syntheses 1147:1155 the highly convergent syntheses 1125:1155 the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites 1125:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 7 74 theme molecule 1584:1591 arg1 portion 1569:1575 the C-terminal portion 1554:1575 the C-terminal portion of the molecule 1554:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 4 75 theme vaccines 848:855 arg1 development 826:836 the development 822:836 the development of cancer vaccines 822:855 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 1 76 dep hormones 213:220 arg1 each 222:225 each 222:225 each 222:225 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 6 77 theme β-hLH 1172:1176 arg1 syntheses 1147:1155 the highly convergent syntheses 1125:1155 the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites 1125:1242 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 4 78 from marker 747:752 arg1 variety 759:765 a variety 757:765 a variety of cancers 757:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 4 78 from marker 747:752 arg1 cancers 770:776 cancers 770:776 cancers 770:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 1 79 theme Human 119:123 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 79 theme Human 119:123 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 79 theme Human 119:123 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 7 80 theme en 1325:1326 arg1 route 1328:1332 a successful double Lansbury glycosylation en route 1282:1332 a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1282:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 81 theme C-terminal 1558:1567 arg1 portion 1569:1575 the C-terminal portion 1554:1575 the C-terminal portion of the molecule 1554:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 2 82 theme N-glycosylation 433:447 arg1 site 449:452 a common N-glycosylation site 424:452 a common N-glycosylation site 424:452 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 0 83 theme gonadotropin 77:88 arg1 β-subunit 26:34 the β-subunit 22:34 the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones 22:116 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 1 84 theme noncovalent 312:322 arg1 heterodimers 324:335 noncovalent heterodimers 312:335 noncovalent heterodimers 312:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 7 85 theme double 1295:1300 arg1 route 1328:1332 a successful double Lansbury glycosylation en route 1282:1332 a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1282:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 86 link O-linked 1410:1417 arg1 cassettes 1439:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 four O-linked glycosyl-amino acid cassettes 1405:1447 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 4 87 theme hCG 732:734 arg1 role 724:727 the role 720:727 the role of hCG as a tumor marker in a variety of cancers 720:776 In addition, the role of hCG as a tumor marker in a variety of cancers has also attracted significant interest for the development of cancer vaccines. 24806200 2 88 gly N-glycosylation 433:447 arg2 site 449:452 a common N-glycosylation site 424:452 a common N-glycosylation site 424:452 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 7 89 gly O-glycosylation 1469:1483 arg2 sites 1485:1489 closely spaced O-glycosylation sites 1454:1489 closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1454:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 2 90 theme sequence 382:389 arg1 similarity 391:400 sequence similarity 382:400 sequence similarity 382:400 Structurally, β-hCG shares a high degree of sequence similarity with β-hLH, including a common N-glycosylation site at the N-terminus but differs mainly in the presence of an extended C-terminal portion incorporating four closely spaced O-linked glycans. 24806200 3 91 theme important 618:626 arg1 roles 628:632 important roles 618:632 important roles 618:632 These glycoproteins play important roles in reproduction and are used clinically in the treatment of infertility. 24806200 5 92 theme biological 964:973 arg1 methods 975:981 biological methods 964:981 biological methods 964:981 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 0 93 theme luteinizing 45:55 arg1 β-subunit 26:34 the β-subunit 22:34 the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones 22:116 Chemical synthesis of the β-subunit of human luteinizing (hLH) and chorionic gonadotropin (hCG) glycoprotein hormones. 24806200 7 94 theme solid-supported 1518:1532 arg1 synthesis 1534:1542 a single, high-yielding solid-supported synthesis 1494:1542 a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule 1494:1591 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 7 95 theme Key 1245:1247 arg1 steps 1249:1253 Key steps 1245:1253 Key steps in these syntheses 1245:1272 Key steps in these syntheses include a successful double Lansbury glycosylation en route to the N-terminal fragment of β-hCG and the sequential installation of four O-linked glycosyl-amino acid cassettes into closely spaced O-glycosylation sites in a single, high-yielding solid-supported synthesis to access the C-terminal portion of the molecule. 24806200 5 96 theme due 942:944 arg1 glycoforms 931:940 glycoforms 931:940 glycoforms due to limitations of biological methods 931:981 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 5 97 gly glycoproteins 1025:1037 arg1 glycoproteins 1025:1037 these glycoproteins 1019:1037 these glycoproteins 1019:1037 In clinical applications, these hormones are administered as mixtures of glycoforms due to limitations of biological methods in producing homogeneous samples of these glycoproteins. 24806200 8 98 theme final 1598:1602 arg1 assembly 1604:1611 The final assembly 1594:1611 The final assembly of the individual glycopeptide fragments 1594:1652 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 99 theme native 1674:1679 arg1 strategy 1699:1706 a stepwise native chemical ligation strategy 1663:1706 a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms 1663:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 8 100 gly glycopeptide 1631:1642 arg2 glycopeptide 1631:1642 glycopeptide 1631:1642 glycopeptide 1631:1642 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 6 101 theme chemical 1068:1075 arg1 synthesis 1077:1085 chemical synthesis 1068:1085 chemical synthesis 1068:1085 Using the powerful tools of chemical synthesis, the work presented herein focuses on the highly convergent syntheses of homogeneous β-hLH and β-hCG bearing model glycans at all native glycosylation sites. 24806200 8 102 theme ligation 1690:1697 arg1 strategy 1699:1706 a stepwise native chemical ligation strategy 1663:1706 a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms 1663:1853 The final assembly of the individual glycopeptide fragments involved a stepwise native chemical ligation strategy to provide the longest and most complex human glycoprotein hormone (β-hCG) as well as its closely related homologue (β-hLH) as discrete glycoforms. 24806200 1 103 theme glycoprotein 200:211 arg1 gonadotropin 171:182 human chorionic gonadotropin 155:182 human chorionic gonadotropin (hCG) 155:188 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 103 theme glycoprotein 200:211 arg1 hormone 137:143 Human luteinizing hormone 119:143 Human luteinizing hormone (hLH) 119:149 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 24806200 1 103 theme glycoprotein 200:211 arg1 hormones 213:220 human glycoprotein hormones 194:220 human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers 194:335 Human luteinizing hormone (hLH) and human chorionic gonadotropin (hCG) are human glycoprotein hormones each consisting of two subunits, an identical α-subunit and a unique β-subunit, that form noncovalent heterodimers. 27216994 7 0 theme HEK293 1331:1336 arg1 cells 1338:1342 HEK293 cells 1331:1342 HEK293 cells that express recombinant GluN1/GluN2B receptors 1331:1390 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 5 1 used used 722:725 arg2 we 719:720 we 719:720 we 719:720 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 9 2 theme central 1734:1740 arg1 system 1750:1755 the central nervous system 1730:1755 the central nervous system 1730:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 4 3 theme GluN1 651:655 arg1 subunits 657:664 GluN1 subunits 651:664 GluN1 subunits 651:664 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 6 4 theme GluN2B 940:945 arg1 subunits 953:960 GluN2B NMDAR subunits 940:960 GluN2B NMDAR subunits 940:960 Our experiments identified 23 lectins that pulled down both the GluN1 and GluN2B NMDAR subunits. 27216994 13 5 gly N-glycosylation 2244:2258 arg2 sites 2260:2264 11 out of 12 predicted N-glycosylation sites 2221:2264 11 out of 12 predicted N-glycosylation sites in GluN1 2221:2273 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 13 5 gly N-glycosylation 2244:2258 arg2 12 2231:2232 12 2231:2232 12 2231:2232 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 10 6 theme ethics 1838:1843 arg1 guidelines 1845:1854 relevant institutional ethics guidelines 1815:1854 relevant institutional ethics guidelines 1815:1854 All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare. 27216994 4 7 gly N-glycosylated 573:586 arg1 NMDARs 550:555 native NMDARs 543:555 native NMDARs 543:555 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 1 8 theme α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic 185:231 arg1 receptors 238:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 14 9 theme therapeutic 2465:2475 arg1 approaches 2477:2486 novel therapeutic approaches 2459:2486 novel therapeutic approaches 2459:2486 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 4 10 theme H 698:698 arg1 sensitivity 700:710 endoglycosidase H sensitivity 682:710 endoglycosidase H sensitivity 682:710 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 13 11 theme predicted 2234:2242 arg1 sites 2260:2264 11 out of 12 predicted N-glycosylation sites 2221:2264 11 out of 12 predicted N-glycosylation sites in GluN1 2221:2273 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 12 from sites 1521:1525 arg1 GluN2B 1576:1581 GluN2B 1576:1581 GluN2B 1576:1581 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 12 from sites 1521:1525 arg1 GluN1 1530:1534 GluN1 1530:1534 GluN1 1530:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 13 gly N-glycosylation 1551:1565 arg2 sites 1567:1571 7 out of 7 N-glycosylation sites 1540:1571 7 out of 7 N-glycosylation sites in GluN2B 1540:1581 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 13 gly N-glycosylation 1551:1565 arg2 7 1549:1549 7 1549:1549 7 1549:1549 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 11 14 theme N-glycan 1931:1938 arg1 composition 1940:1950 the N-glycan composition 1927:1950 the N-glycan composition of NMDA receptors (NMDARs) 1927:1977 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 12 15 theme different 2122:2130 arg1 lectins 2132:2138 23 different lectins 2119:2138 23 different lectins that have unique specificities for glycan structures 2119:2191 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 5 16 theme complete 729:736 arg1 panel 738:742 a complete panel 727:742 a complete panel of lectins 727:753 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 10 17 theme animal 1899:1904 arg1 welfare 1906:1912 animal welfare 1899:1912 animal welfare 1899:1912 All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare. 27216994 7 18 theme granule 1084:1090 arg1 cells 1092:1096 pre-incubating cerebellar granule cells 1058:1096 pre-incubating cerebellar granule cells with the AAL, WGA, or ConA 1058:1123 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 8 19 theme mutant 1408:1413 arg1 subunits 1432:1439 mutant recombinant GluN subunits 1408:1439 mutant recombinant GluN subunits expressed in HEK293 cells 1408:1465 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 20 used occupied 1587:1594 arg2 sites 1521:1525 11 out of 12 predicted N-glycosylation sites 1482:1525 11 out of 12 predicted N-glycosylation sites in GluN1 1482:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 20 used occupied 1587:1594 arg2 sites 1567:1571 7 out of 7 N-glycosylation sites 1540:1571 7 out of 7 N-glycosylation sites in GluN2B 1540:1581 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 7 21 theme pre-incubating 1058:1071 arg1 cells 1092:1096 pre-incubating cerebellar granule cells 1058:1096 pre-incubating cerebellar granule cells with the AAL, WGA, or ConA 1058:1123 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 8 22 theme GluN 1427:1430 arg1 subunits 1432:1439 mutant recombinant GluN subunits 1408:1439 mutant recombinant GluN subunits expressed in HEK293 cells 1408:1465 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 0 23 theme receptor 75:82 arg1 subunits 84:91 NMDA receptor subunits 70:91 NMDA receptor subunits 70:91 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 2 24 theme receptor 298:305 arg1 types 307:311 Both receptor types 293:311 Both receptor types 293:311 Both receptor types are comprised of several subunits that are post-translationally modified by N-glycosylation. 27216994 1 25 theme ionotropic 153:162 arg1 receptors 174:182 ionotropic glutamate receptors 153:182 ionotropic glutamate receptors 153:182 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 13 26 from sites 2260:2264 arg1 GluN2B 2315:2320 GluN2B 2315:2320 GluN2B 2315:2320 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 13 26 from sites 2260:2264 arg1 GluN1 2269:2273 GluN1 2269:2273 GluN1 2269:2273 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 14 27 theme potential 2418:2426 arg1 targets 2428:2434 potential targets 2418:2434 potential targets for the development of novel therapeutic approaches 2418:2486 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 4 28 used used 509:512 arg2 we 506:507 we 506:507 we 506:507 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 13 29 used occupied 2326:2333 arg2 sites 2260:2264 11 out of 12 predicted N-glycosylation sites 2221:2264 11 out of 12 predicted N-glycosylation sites in GluN1 2221:2273 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 13 29 used occupied 2326:2333 arg2 7 2279:2279 7 2279:2279 7 2279:2279 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 1 30 theme receptors 174:182 arg1 types 144:148 two major types 134:148 two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs) 134:290 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 7 31 with cells 1092:1096 arg1 WGA 1112:1114 WGA 1112:1114 WGA 1112:1114 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 7 31 with cells 1092:1096 arg1 ConA 1120:1123 ConA 1120:1123 ConA 1120:1123 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 7 31 with cells 1092:1096 arg1 AAL 1107:1109 AAL 1107:1109 AAL 1107:1109 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 8 32 theme HEK293 1454:1459 arg1 cells 1461:1465 HEK293 cells 1454:1465 HEK293 cells 1454:1465 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 33 from sites 1567:1571 arg1 GluN2B 1576:1581 GluN2B 1576:1581 GluN2B 1576:1581 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 33 from sites 1567:1571 arg1 GluN1 1530:1534 GluN1 1530:1534 GluN1 1530:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 9 34 theme new 1629:1631 arg1 insight 1633:1639 new insight 1629:1639 new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system 1629:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 11 35 theme lectin-based 2010:2021 arg1 biochemistry 2023:2034 lectin-based biochemistry 2010:2034 lectin-based biochemistry 2010:2034 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 7 36 theme F. 1276:1277 arg1 effects 1303:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 7 37 theme electrophysiological 984:1003 arg1 analysis 1005:1012 an electrophysiological analysis 981:1012 an electrophysiological analysis 981:1012 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 9 38 from function 1700:1707 arg1 system 1750:1755 the central nervous system 1730:1755 the central nervous system 1730:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 4 39 theme native 543:548 arg1 NMDARs 550:555 native NMDARs 543:555 native NMDARs 543:555 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 14 40 theme glycan 2378:2383 arg1 composition 2385:2395 the glycan composition 2374:2395 the glycan composition of NMDARs 2374:2405 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 0 41 theme Biochemical 0:10 arg1 characterization 37:52 Biochemical and electrophysiological characterization 0:52 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 0:92 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 2 42 mod modified 377:384 arg1 subunits 338:345 several subunits 330:345 several subunits that are post-translationally modified by N-glycosylation 330:403 Both receptor types are comprised of several subunits that are post-translationally modified by N-glycosylation. 27216994 2 42 mod modified 377:384 arg3 N-glycosylation 389:403 N-glycosylation 389:403 N-glycosylation 389:403 Both receptor types are comprised of several subunits that are post-translationally modified by N-glycosylation. 27216994 10 43 theme animal 1762:1767 arg1 experiments 1769:1779 All animal experiments 1758:1779 All animal experiments 1758:1779 All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare. 27216994 0 44 theme electrophysiological 16:35 arg1 characterization 37:52 Biochemical and electrophysiological characterization 0:52 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 0:92 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 4 45 theme NMDAR 617:621 arg1 subunit 630:636 the NMDAR GluN2B subunit 613:636 the NMDAR GluN2B subunit 613:636 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 13 46 gly 7 N-glycosylation 2288:2304 arg2 sites 2306:2310 7 N-glycosylation sites 2288:2310 7 N-glycosylation sites 2288:2310 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 5 47 theme cultured 831:838 arg1 cells 859:863 cultured cerebellar granule cells 831:863 cultured cerebellar granule cells 831:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 3 48 theme receptor 464:471 arg1 types 473:477 these receptor types 458:477 these receptor types 458:477 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 8 49 theme predicted 1495:1503 arg1 sites 1521:1525 11 out of 12 predicted N-glycosylation sites 1482:1525 11 out of 12 predicted N-glycosylation sites in GluN1 1482:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 11 50 theme receptors 1960:1968 arg1 composition 1940:1950 the N-glycan composition 1927:1950 the N-glycan composition of NMDA receptors (NMDARs) 1927:1977 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 5 51 theme granule 851:857 arg1 cells 859:863 cultured cerebellar granule cells 831:863 cultured cerebellar granule cells 831:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 7 52 theme GluN1/GluN2B 1369:1380 arg1 receptors 1382:1390 recombinant GluN1/GluN2B receptors 1357:1390 recombinant GluN1/GluN2B receptors 1357:1390 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 10 53 theme institutional 1824:1836 arg1 guidelines 1845:1854 relevant institutional ethics guidelines 1815:1854 relevant institutional ethics guidelines 1815:1854 All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare. 27216994 0 54 theme NMDA 70:73 arg1 subunits 84:91 NMDA receptor subunits 70:91 NMDA receptor subunits 70:91 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 9 55 theme receptors 1717:1725 arg1 function 1700:1707 the function 1696:1707 the function of NMDA receptors in the central nervous system 1696:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 7 56 theme lectin-mediated 1176:1190 arg1 effect 1192:1197 this lectin-mediated effect 1171:1197 this lectin-mediated effect 1171:1197 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 8 57 gly N-glycosylation 1505:1519 arg2 sites 1521:1525 11 out of 12 predicted N-glycosylation sites 1482:1525 11 out of 12 predicted N-glycosylation sites in GluN1 1482:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 8 57 gly N-glycosylation 1505:1519 arg2 12 1492:1493 12 1492:1493 12 1492:1493 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 7 58 dep AAL 1107:1109 arg1 the 1103:1105 the 1103:1105 the 1103:1105 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 1 59 contain contain 126:132 arg2 synapses 117:124 excitatory synapses 106:124 excitatory synapses 106:124 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 59 contain contain 126:132 arg2 types 144:148 two major types 134:148 two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs) 134:290 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 59 contain contain 126:132 arg1 mammals 97:103 mammals 97:103 mammals 97:103 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 59 contain contain 126:132 arg1 synapses 117:124 excitatory synapses 106:124 excitatory synapses 106:124 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 6 60 theme NMDAR 947:951 arg1 subunits 953:960 GluN2B NMDAR subunits 940:960 GluN2B NMDAR subunits 940:960 Our experiments identified 23 lectins that pulled down both the GluN1 and GluN2B NMDAR subunits. 27216994 9 61 theme nervous 1742:1748 arg1 system 1750:1755 the central nervous system 1730:1755 the central nervous system 1730:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 7 62 theme biophysical 1147:1157 arg1 properties 1159:1168 the receptor's biophysical properties 1132:1168 the receptor's biophysical properties 1132:1168 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 2 63 theme several 330:336 arg1 subunits 338:345 several subunits 330:345 several subunits that are post-translationally modified by N-glycosylation 330:403 Both receptor types are comprised of several subunits that are post-translationally modified by N-glycosylation. 27216994 14 64 theme approaches 2477:2486 arg1 development 2444:2454 the development 2440:2454 the development of novel therapeutic approaches 2440:2486 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 1 65 theme acid 233:236 arg1 receptors 238:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 12 66 theme cerebellar 2086:2095 arg1 NMDARs 2097:2102 cerebellar NMDARs 2086:2102 cerebellar NMDARs 2086:2102 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 5 67 theme lectins 747:753 arg1 panel 738:742 a complete panel 727:742 a complete panel of lectins 727:753 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 14 68 theme novel 2459:2463 arg1 approaches 2477:2486 novel therapeutic approaches 2459:2486 novel therapeutic approaches 2459:2486 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 12 69 contain have 2145:2148 arg1 lectins 2132:2138 23 different lectins 2119:2138 23 different lectins that have unique specificities for glycan structures 2119:2191 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 12 69 contain have 2145:2148 arg2 specificities 2157:2169 unique specificities 2150:2169 unique specificities for glycan structures 2150:2191 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 11 70 gly deglycosylating 1985:1999 arg0 enzymes 2001:2007 deglycosylating enzymes 1985:2007 deglycosylating enzymes 1985:2007 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 4 71 theme endoglycosidase 682:696 arg1 sensitivity 700:710 endoglycosidase H sensitivity 682:710 endoglycosidase H sensitivity 682:710 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 13 72 theme N-glycosylation 2244:2258 arg1 sites 2260:2264 11 out of 12 predicted N-glycosylation sites 2221:2264 11 out of 12 predicted N-glycosylation sites in GluN1 2221:2273 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 5 73 theme glycan 772:777 arg1 composition 779:789 the glycan composition 768:789 the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells 768:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 5 74 from composition 779:789 arg1 cells 859:863 cultured cerebellar granule cells 831:863 cultured cerebellar granule cells 831:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 5 74 from composition 779:789 arg1 tissue 820:825 cerebellar tissue 809:825 cerebellar tissue 809:825 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 8 75 theme N-glycosylation 1551:1565 arg1 sites 1567:1571 7 out of 7 N-glycosylation sites 1540:1571 7 out of 7 N-glycosylation sites in GluN2B 1540:1581 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 1 76 theme major 138:142 arg1 types 144:148 two major types 134:148 two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs) 134:290 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 3 77 attach attached 446:453 arg1 types 473:477 these receptor types 458:477 these receptor types 458:477 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 3 77 attach attached 446:453 arg2 N-glycans 427:435 the precise N-glycans 415:435 the precise N-glycans that are attached to these receptor types 415:477 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 3 77 attach attached 446:453 arg2 unknown 491:497 unknown 491:497 unknown 491:497 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 7 78 theme cerebellar 1073:1082 arg1 cells 1092:1096 pre-incubating cerebellar granule cells 1058:1096 pre-incubating cerebellar granule cells with the AAL, WGA, or ConA 1058:1123 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 0 79 from characterization 37:52 arg1 subunits 84:91 NMDA receptor subunits 70:91 NMDA receptor subunits 70:91 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 8 80 theme recombinant 1415:1425 arg1 subunits 1432:1439 mutant recombinant GluN subunits 1408:1439 mutant recombinant GluN subunits expressed in HEK293 cells 1408:1465 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 7 81 theme lectin-mediated 1287:1301 arg1 effects 1303:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 5 82 theme cerebellar 809:818 arg1 tissue 820:825 cerebellar tissue 809:825 cerebellar tissue 809:825 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 1 83 theme glutamate 164:172 arg1 receptors 174:182 ionotropic glutamate receptors 153:182 ionotropic glutamate receptors 153:182 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 9 84 dep role 1650:1653 arg1 plays 1676:1680 plays 1676:1680 plays in regulating the function of NMDA receptors in the central nervous system 1676:1755 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 7 85 theme Similar 1279:1285 arg1 effects 1303:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 8 86 theme N-glycosylation 1505:1519 arg1 sites 1521:1525 11 out of 12 predicted N-glycosylation sites 1482:1525 11 out of 12 predicted N-glycosylation sites in GluN1 1482:1534 Finally, using mutant recombinant GluN subunits expressed in HEK293 cells, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 1 87 dep types 144:148 arg1 NMDARs 284:289 NMDARs 284:289 NMDARs 284:289 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 87 dep types 144:148 arg1 receptors 238:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors 185:246 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 87 dep types 144:148 arg1 receptors 273:281 N-methyl-d-aspartate receptors 252:281 N-methyl-d-aspartate receptors (NMDARs) 252:290 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 14 88 theme NMDARs 2400:2405 arg1 composition 2385:2395 the glycan composition 2374:2395 the glycan composition of NMDARs 2374:2405 These data shed light on the glycan composition of NMDARs, revealing potential targets for the development of novel therapeutic approaches. 27216994 11 89 theme NMDA 1955:1958 arg1 receptors 1960:1968 NMDA receptors 1955:1968 NMDA receptors (NMDARs) 1955:1977 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 11 89 theme NMDA 1955:1958 arg1 NMDARs 1971:1976 NMDARs 1971:1976 NMDARs 1971:1976 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 7 90 theme peptide-N-glycosidase 1254:1274 arg1 effects 1303:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 peptide-N-glycosidase F. Similar lectin-mediated effects 1254:1309 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 7 91 theme representative 1020:1033 arg1 lectins 1035:1041 representative lectins 1020:1041 representative lectins 1020:1041 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 3 92 theme precise 419:425 arg1 unknown 491:497 unknown 491:497 unknown 491:497 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 3 92 theme precise 419:425 arg1 N-glycans 427:435 the precise N-glycans 415:435 the precise N-glycans that are attached to these receptor types 415:477 However, the precise N-glycans that are attached to these receptor types are largely unknown. 27216994 1 93 theme excitatory 106:115 arg1 synapses 117:124 excitatory synapses 106:124 excitatory synapses 106:124 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 4 94 theme GluN2B 623:628 arg1 subunit 630:636 the NMDAR GluN2B subunit 613:636 the NMDAR GluN2B subunit 613:636 Here, we used biochemistry to confirm that native NMDARs are extensively N-glycosylated; moreover, we found that the NMDAR GluN2B subunit differs from GluN1 subunits with respect to endoglycosidase H sensitivity. 27216994 0 95 theme N-glycans 57:65 arg1 characterization 37:52 Biochemical and electrophysiological characterization 0:52 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 0:92 Biochemical and electrophysiological characterization of N-glycans on NMDA receptor subunits. 27216994 12 96 theme glycan 2175:2180 arg1 structures 2182:2191 glycan structures 2175:2191 glycan structures 2175:2191 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 5 97 theme NMDARs 794:799 arg1 composition 779:789 the glycan composition 768:789 the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells 768:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 5 98 theme cerebellar 840:849 arg1 cells 859:863 cultured cerebellar granule cells 831:863 cultured cerebellar granule cells 831:863 Next, we used a complete panel of lectins to determine the glycan composition of NMDARs in both cerebellar tissue and cultured cerebellar granule cells. 27216994 7 99 theme recombinant 1357:1367 arg1 receptors 1382:1390 recombinant GluN1/GluN2B receptors 1357:1390 recombinant GluN1/GluN2B receptors 1357:1390 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 7 100 gly deglycosylated 1234:1247 arg1 cells 1223:1227 the cells 1219:1227 the cells 1219:1227 We then performed an electrophysiological analysis using representative lectins and found that pre-incubating cerebellar granule cells with the AAL, WGA, or ConA alters the receptor's biophysical properties; this lectin-mediated effect was eliminated when the cells were deglycosylated with peptide-N-glycosidase F. Similar lectin-mediated effects were observed using HEK293 cells that express recombinant GluN1/GluN2B receptors. 27216994 1 101 theme N-methyl-d-aspartate 252:271 arg1 NMDARs 284:289 NMDARs 284:289 NMDARs 284:289 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 1 101 theme N-methyl-d-aspartate 252:271 arg1 receptors 273:281 N-methyl-d-aspartate receptors 252:281 N-methyl-d-aspartate receptors (NMDARs) 252:290 In mammals, excitatory synapses contain two major types of ionotropic glutamate receptors: α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors and N-methyl-d-aspartate receptors (NMDARs). 27216994 11 102 theme deglycosylating 1985:1999 arg1 enzymes 2001:2007 deglycosylating enzymes 1985:2007 deglycosylating enzymes 1985:2007 We examined the N-glycan composition of NMDA receptors (NMDARs) using deglycosylating enzymes, lectin-based biochemistry, and electrophysiology. 27216994 13 103 theme 7 N-glycosylation 2288:2304 arg1 sites 2306:2310 7 N-glycosylation sites 2288:2310 7 N-glycosylation sites 2288:2310 Furthermore, we found that 11 out of 12 predicted N-glycosylation sites in GluN1 and 7 out of 7 N-glycosylation sites in GluN2B are occupied by N-glycans. 27216994 12 104 theme unique 2150:2155 arg1 specificities 2157:2169 unique specificities 2150:2169 unique specificities for glycan structures 2150:2191 Our results revealed that cerebellar NMDARs associate with 23 different lectins that have unique specificities for glycan structures. 27216994 9 105 theme NMDA 1712:1715 arg1 receptors 1717:1725 NMDA receptors 1712:1725 NMDA receptors 1712:1725 These data provide new insight into the role that N-glycosylation plays in regulating the function of NMDA receptors in the central nervous system. 27216994 10 106 theme relevant 1815:1822 arg1 guidelines 1845:1854 relevant institutional ethics guidelines 1815:1854 relevant institutional ethics guidelines 1815:1854 All animal experiments were performed in accordance with relevant institutional ethics guidelines and regulations with respect to protecting animal welfare. 26439794 7 0 contain containing 1019:1028 arg1 type 1014:1017 high-mannose type 1001:1017 high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides 1001:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 0 contain containing 1019:1028 arg2 M9 1053:1054 M9 1053:1054 M9 1053:1054 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 0 contain containing 1019:1028 arg2 oligosaccharides 1095:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 0 contain containing 1019:1028 arg2 residues 1043:1050 nine mannose residues 1030:1050 nine mannose residues (M9) 1030:1055 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 0 contain containing 1019:1028 arg1 forms 966:970 The most frequently occurring forms 936:970 The most frequently occurring forms at N-glycosylation site 936:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 8 1 theme fucosylated 1212:1222 arg1 types 1238:1242 fucosylated pauci-mannose types 1212:1242 fucosylated pauci-mannose types 1212:1242 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 3 2 gly glycopeptide 417:428 arg2 glycopeptide 417:428 a glycopeptide enrichment method 415:446 a glycopeptide enrichment method 415:446 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 7 3 theme high-mannose 1001:1012 arg1 type 1014:1017 high-mannose type 1001:1017 high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides 1001:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 3 theme high-mannose 1001:1012 arg1 forms 966:970 The most frequently occurring forms 936:970 The most frequently occurring forms at N-glycosylation site 936:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 3 4 theme human 509:513 arg1 glycoproteomics 521:535 human serum glycoproteomics 509:535 human serum glycoproteomics 509:535 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 8 5 theme lysosome 1265:1272 arg1 proteins 1274:1281 lysosome proteins 1265:1281 lysosome proteins 1265:1281 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 5 6 theme fibroblasts 715:725 arg1 fraction 736:743 the human fetal lung fibroblasts membrane fraction 694:743 the human fetal lung fibroblasts membrane fraction 694:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 2 7 from intensity 330:338 arg1 existence 364:372 the existence 360:372 the existence of surfactants 360:387 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 3 8 theme serum 515:519 arg1 glycoproteomics 521:535 human serum glycoproteomics 509:535 human serum glycoproteomics 509:535 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 2 9 contain has 242:244 arg2 trouble 246:252 trouble 246:252 trouble in mass spectrometric glycoproteomics of membrane fraction 246:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 2 9 contain has 242:244 arg1 one 238:240 one 238:240 one 238:240 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 9 10 theme new 1304:1306 arg1 insights 1308:1315 new insights 1304:1315 new insights into transport, biosynthesis, and degradation of glycoproteins 1304:1378 Our method provides new insights into transport, biosynthesis, and degradation of glycoproteins. 26439794 9 11 theme glycoproteins 1366:1378 arg1 biosynthesis 1333:1344 biosynthesis 1333:1344 biosynthesis 1333:1344 Our method provides new insights into transport, biosynthesis, and degradation of glycoproteins. 26439794 9 11 theme glycoproteins 1366:1378 arg1 degradation 1351:1361 degradation 1351:1361 degradation 1351:1361 Our method provides new insights into transport, biosynthesis, and degradation of glycoproteins. 26439794 9 11 theme glycoproteins 1366:1378 arg1 transport 1322:1330 transport 1322:1330 transport 1322:1330 Our method provides new insights into transport, biosynthesis, and degradation of glycoproteins. 26439794 5 12 theme glycoproteomic 667:680 arg1 approach 682:689 The glycoproteomic approach 663:689 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction 663:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 8 13 located found 1249:1253 arg2 high-mannose 1192:1203 high-mannose 1192:1203 high-mannose 1192:1203 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 13 located found 1249:1253 arg1 proteins 1274:1281 lysosome proteins 1265:1281 lysosome proteins 1265:1281 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 13 located found 1249:1253 arg2 types 1238:1242 fucosylated pauci-mannose types 1212:1242 fucosylated pauci-mannose types 1212:1242 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 13 located found 1249:1253 arg2 complex-type 1163:1174 complex-type 1163:1174 complex-type 1163:1174 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 13 located found 1249:1253 arg1 ER 1258:1259 ER 1258:1259 ER 1258:1259 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 13 located found 1249:1253 arg2 N-glycans 1132:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 6 14 from features 888:895 arg1 sites 903:907 41 sites 900:907 41 sites 900:907 According to the existing databases, the structural features on 41 sites are previously unreported. 26439794 1 15 theme tumor 113:117 arg1 N-glycosylations 78:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 1 15 theme tumor 113:117 arg1 markers 119:125 tumor markers 113:125 tumor markers 113:125 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 1 16 theme aberrant 69:76 arg1 N-glycosylations 78:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 1 16 theme aberrant 69:76 arg1 markers 119:125 tumor markers 113:125 tumor markers 113:125 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 3 17 theme enrichment 430:439 arg1 method 441:446 a glycopeptide enrichment method 415:446 a glycopeptide enrichment method 415:446 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 7 18 gly monosialo-fucosylated 1061:1081 arg1 oligosaccharides 1095:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 2 19 theme mass 257:260 arg1 glycoproteomics 276:290 mass spectrometric glycoproteomics 257:290 mass spectrometric glycoproteomics of membrane fraction 257:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 1 20 theme novel 171:175 arg1 markers 187:193 novel diagnosis markers 171:193 novel diagnosis markers 171:193 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 5 21 theme glycoforms 788:797 arg1 identification 761:774 the identification 757:774 the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins 757:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 8 22 theme Several 1113:1119 arg1 complex-type 1163:1174 complex-type 1163:1174 complex-type 1163:1174 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 22 theme Several 1113:1119 arg1 N-glycans 1132:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 22 theme Several 1113:1119 arg1 high-mannose 1192:1203 high-mannose 1192:1203 high-mannose 1192:1203 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 7 23 theme occurring 956:964 arg1 type 1014:1017 high-mannose type 1001:1017 high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides 1001:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 23 theme occurring 956:964 arg1 forms 966:970 The most frequently occurring forms 936:970 The most frequently occurring forms at N-glycosylation site 936:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 24 theme monosialo-fucosylated 1061:1081 arg1 oligosaccharides 1095:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 1 25 theme diagnosis 177:185 arg1 markers 187:193 novel diagnosis markers 171:193 novel diagnosis markers 171:193 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 0 26 theme Membrane 0:7 arg1 glycoproteomics 9:23 Membrane glycoproteomics 0:23 Membrane glycoproteomics of fetal lung fibroblasts 0:49 Membrane glycoproteomics of fetal lung fibroblasts using LC/MS. 26439794 2 27 theme surfactants 377:387 arg1 existence 364:372 the existence 360:372 the existence of surfactants 360:387 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 0 28 theme fetal 28:32 arg1 fibroblasts 39:49 fetal lung fibroblasts 28:49 fetal lung fibroblasts 28:49 Membrane glycoproteomics of fetal lung fibroblasts using LC/MS. 26439794 5 29 theme lung 710:713 arg1 fibroblasts 715:725 human fetal lung fibroblasts 698:725 the human fetal lung fibroblasts membrane fraction 694:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 7 30 theme most 940:943 arg1 type 1014:1017 high-mannose type 1001:1017 high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides 1001:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 30 theme most 940:943 arg1 forms 966:970 The most frequently occurring forms 936:970 The most frequently occurring forms at N-glycosylation site 936:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 2 31 theme fraction 304:311 arg1 glycoproteomics 276:290 mass spectrometric glycoproteomics 257:290 mass spectrometric glycoproteomics of membrane fraction 257:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 7 32 theme N-glycosylation 975:989 arg1 site 991:994 N-glycosylation site 975:994 N-glycosylation site 975:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 33 theme biantennary 1083:1093 arg1 oligosaccharides 1095:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 monosialo-fucosylated biantennary oligosaccharides 1061:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 2 34 theme spectrometric 262:274 arg1 glycoproteomics 276:290 mass spectrometric glycoproteomics 257:290 mass spectrometric glycoproteomics of membrane fraction 257:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 6 35 theme structural 877:886 arg1 features 888:895 the structural features 873:895 the structural features on 41 sites 873:907 According to the existing databases, the structural features on 41 sites are previously unreported. 26439794 2 36 theme membrane 295:302 arg1 fraction 304:311 membrane fraction 295:311 membrane fraction 295:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 0 37 theme fibroblasts 39:49 arg1 glycoproteomics 9:23 Membrane glycoproteomics 0:23 Membrane glycoproteomics of fetal lung fibroblasts 0:49 Membrane glycoproteomics of fetal lung fibroblasts using LC/MS. 26439794 5 38 theme membrane 727:734 arg1 fraction 736:743 the human fetal lung fibroblasts membrane fraction 694:743 the human fetal lung fibroblasts membrane fraction 694:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 39 from identification 761:774 arg1 sites 805:809 63 sites 802:809 63 sites of the 44 glycoproteins 802:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 39 from identification 761:774 arg1 glycoproteins 821:833 the 44 glycoproteins 814:833 the 44 glycoproteins 814:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 8 40 theme unexpected 1121:1130 arg1 complex-type 1163:1174 complex-type 1163:1174 complex-type 1163:1174 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 40 theme unexpected 1121:1130 arg1 N-glycans 1132:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans 1113:1140 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 8 40 theme unexpected 1121:1130 arg1 high-mannose 1192:1203 high-mannose 1192:1203 high-mannose 1192:1203 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 2 41 from trouble 246:252 arg1 glycoproteomics 276:290 mass spectrometric glycoproteomics 257:290 mass spectrometric glycoproteomics of membrane fraction 257:311 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 7 42 from site 991:994 arg1 type 1014:1017 high-mannose type 1001:1017 high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides 1001:1110 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 42 from site 991:994 arg1 forms 966:970 The most frequently occurring forms 936:970 The most frequently occurring forms at N-glycosylation site 936:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 0 43 theme lung 34:37 arg1 fibroblasts 39:49 fetal lung fibroblasts 28:49 fetal lung fibroblasts 28:49 Membrane glycoproteomics of fetal lung fibroblasts using LC/MS. 26439794 3 44 theme glycopeptide 417:428 arg1 method 441:446 a glycopeptide enrichment method 415:446 a glycopeptide enrichment method 415:446 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 5 45 theme glycoproteins 821:833 arg1 sites 805:809 63 sites 802:809 63 sites of the 44 glycoproteins 802:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 45 theme glycoproteins 821:833 arg1 glycoproteins 821:833 the 44 glycoproteins 814:833 the 44 glycoproteins 814:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 1 46 theme drug 210:213 arg1 developments 215:226 drug developments 210:226 drug developments 210:226 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 7 47 gly N-glycosylation 975:989 arg2 site 991:994 N-glycosylation site 975:994 N-glycosylation site 975:994 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 8 48 gly fucosylated 1212:1222 arg1 types 1238:1242 fucosylated pauci-mannose types 1212:1242 fucosylated pauci-mannose types 1212:1242 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 26439794 2 49 theme glycopeptides 343:355 arg1 intensity 330:338 lower intensity 324:338 lower intensity of glycopeptides in the existence of surfactants 324:387 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 2 50 gly glycopeptides 343:355 arg2 glycopeptides 343:355 glycopeptides 343:355 glycopeptides 343:355 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 6 51 theme existing 853:860 arg1 databases 862:870 the existing databases 849:870 the existing databases 849:870 According to the existing databases, the structural features on 41 sites are previously unreported. 26439794 5 52 gly glycoproteins 821:833 arg1 glycoproteins 821:833 the 44 glycoproteins 814:833 the 44 glycoproteins 814:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 1 53 used used 105:108 arg2 N-glycosylations 78:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations 64:93 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 1 53 used used 105:108 arg2 markers 119:125 tumor markers 113:125 tumor markers 113:125 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 5 54 theme human 698:702 arg1 fibroblasts 715:725 human fetal lung fibroblasts 698:725 the human fetal lung fibroblasts membrane fraction 694:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 55 from glycoforms 788:797 arg1 sites 805:809 63 sites 802:809 63 sites of the 44 glycoproteins 802:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 55 from glycoforms 788:797 arg1 glycoproteins 821:833 the 44 glycoproteins 814:833 the 44 glycoproteins 814:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 7 56 theme mannose 1035:1041 arg1 M9 1053:1054 M9 1053:1054 M9 1053:1054 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 7 56 theme mannose 1035:1041 arg1 residues 1043:1050 nine mannose residues 1030:1050 nine mannose residues (M9) 1030:1055 The most frequently occurring forms at N-glycosylation site were high-mannose type containing nine mannose residues (M9) and monosialo-fucosylated biantennary oligosaccharides. 26439794 9 57 gly glycoproteins 1366:1378 arg1 glycoproteins 1366:1378 glycoproteins 1366:1378 glycoproteins 1366:1378 Our method provides new insights into transport, biosynthesis, and degradation of glycoproteins. 26439794 1 58 theme developments 215:226 arg1 targets 199:205 targets 199:205 targets of drug developments 199:226 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 1 58 theme developments 215:226 arg1 markers 187:193 novel diagnosis markers 171:193 novel diagnosis markers 171:193 Some aberrant N-glycosylations are being used as tumor markers, and glycoproteomics is expected to provide novel diagnosis markers and targets of drug developments. 26439794 5 59 theme fetal 704:708 arg1 fibroblasts 715:725 human fetal lung fibroblasts 698:725 the human fetal lung fibroblasts membrane fraction 694:743 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 2 60 theme lower 324:328 arg1 intensity 330:338 lower intensity 324:338 lower intensity of glycopeptides in the existence of surfactants 324:387 However, one has trouble in mass spectrometric glycoproteomics of membrane fraction because of lower intensity of glycopeptides in the existence of surfactants. 26439794 4 61 theme membrane 637:644 arg1 glycoproteomics 646:660 membrane glycoproteomics 637:660 membrane glycoproteomics 637:660 In this study, we confirmed that this method is useful to remove the surfactants and applicable to membrane glycoproteomics. 26439794 3 62 theme acetone 451:457 arg1 precipitation 459:471 acetone precipitation 451:471 acetone precipitation 451:471 Previously, we developed a glycopeptide enrichment method by acetone precipitation, and it was successfully applied to human serum glycoproteomics. 26439794 5 63 from sites 805:809 arg1 identification 761:774 the identification 757:774 the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins 757:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 64 gly glycoforms 788:797 arg2 glycoproteins 821:833 the 44 glycoproteins 814:833 the 44 glycoproteins 814:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 5 64 gly glycoforms 788:797 arg2 sites 805:809 63 sites 802:809 63 sites of the 44 glycoproteins 802:833 The glycoproteomic approach to the human fetal lung fibroblasts membrane fraction resulted in the identification of over 272 glycoforms on 63 sites of the 44 glycoproteins. 26439794 8 65 theme pauci-mannose 1224:1236 arg1 types 1238:1242 fucosylated pauci-mannose types 1212:1242 fucosylated pauci-mannose types 1212:1242 Several unexpected N-glycans, such as fucosylated complex-type and fucosylated high-mannose and/or fucosylated pauci-mannose types were found in ER and lysosome proteins. 25053492 4 0 with fragments 797:805 arg1 proteases 812:820 proteases 812:820 proteases 812:820 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 3 1 theme most 669:672 arg1 modifications 692:704 most posttranslational modifications 669:704 most posttranslational modifications 669:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 4 2 theme digestion 780:788 arg1 purification 743:754 purification 743:754 purification 743:754 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 2 theme digestion 780:788 arg1 expression 728:737 expression 728:737 expression 728:737 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 3 3 from sites 621:625 arg1 Notch 630:634 Notch 630:634 Notch 630:634 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 3 3 from sites 621:625 arg1 modifications 593:605 modifications 593:605 modifications at individual sites on Notch that are generally applicable to most posttranslational modifications 593:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 3 4 theme posttranslational 674:690 arg1 modifications 692:704 most posttranslational modifications 669:704 most posttranslational modifications 669:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 4 5 theme ECD 765:767 arg1 fragments 769:777 Notch ECD fragments 759:777 Notch ECD fragments 759:777 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 6 theme identification 865:878 arg1 purification 743:754 purification 743:754 purification 743:754 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 6 theme identification 865:878 arg1 expression 728:737 expression 728:737 expression 728:737 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 7 theme Notch 759:763 arg1 fragments 769:777 Notch ECD fragments 759:777 Notch ECD fragments 759:777 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 8 dep expression 728:737 arg1 the 724:726 the 724:726 the 724:726 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 2 9 gly N-glycosylation 256:270 arg1 O-glycosylation 300:314 O-glycosylation 300:314 O-glycosylation (O-fucose, O-glucose, and O-GlcNAc) 300:350 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 4 10 theme mass 837:840 arg1 analysis 851:858 mass spectral analysis 837:858 mass spectral analysis 837:858 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 3 11 dep structure 460:468 arg1 the 456:458 the 456:458 the 456:458 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 2 12 theme intracellular 363:375 arg1 domain 377:382 the intracellular domain 359:382 the intracellular domain 359:382 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 4 13 theme fragments 769:777 arg1 purification 743:754 purification 743:754 purification 743:754 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 13 theme fragments 769:777 arg1 expression 728:737 expression 728:737 expression 728:737 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 2 14 theme O-glycosylation 300:314 arg1 types 291:295 at least three types 276:295 at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc) 276:350 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 14 theme O-glycosylation 300:314 arg1 N-glycosylation 256:270 N-glycosylation 256:270 N-glycosylation 256:270 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 4 15 theme spectral 842:849 arg1 analysis 851:858 mass spectral analysis 837:858 mass spectral analysis 837:858 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 3 16 theme semiquantitative 538:553 arg1 methods 569:575 semiquantitative mass spectral methods 538:575 semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications 538:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 0 17 theme modification 32:43 arg1 status 45:50 the posttranslational modification status 10:50 the posttranslational modification status of Notch 10:59 Analyzing the posttranslational modification status of Notch using mass spectrometry. 25053492 4 18 theme fragments 797:805 arg1 identification 865:878 identification 865:878 identification of peptides modified with O-glycans using mass spectrometry 865:938 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 18 theme fragments 797:805 arg1 digestion 780:788 digestion 780:788 digestion of the fragments with proteases to prepare for mass spectral analysis 780:858 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 18 theme fragments 797:805 arg1 fragments 769:777 Notch ECD fragments 759:777 Notch ECD fragments 759:777 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 19 theme mass 922:925 arg1 spectrometry 927:938 mass spectrometry 922:938 mass spectrometry 922:938 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 3 20 theme spectral 560:567 arg1 methods 569:575 semiquantitative mass spectral methods 538:575 semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications 538:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 1 21 theme Notch 192:196 arg1 function 198:205 Notch function 192:205 Notch function 192:205 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 0 22 theme posttranslational 14:30 arg1 status 45:50 the posttranslational modification status 10:50 the posttranslational modification status of Notch 10:59 Analyzing the posttranslational modification status of Notch using mass spectrometry. 25053492 1 23 theme multiple 107:114 arg1 types 116:120 multiple types 107:120 multiple types 107:120 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 3 24 theme mass 555:558 arg1 methods 569:575 semiquantitative mass spectral methods 538:575 semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications 538:704 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 0 25 theme Notch 55:59 arg1 status 45:50 the posttranslational modification status 10:50 the posttranslational modification status of Notch 10:59 Analyzing the posttranslational modification status of Notch using mass spectrometry. 25053492 4 26 theme peptides 883:890 arg1 identification 865:878 identification 865:878 identification of peptides modified with O-glycans using mass spectrometry 865:938 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 26 theme peptides 883:890 arg1 digestion 780:788 digestion 780:788 digestion of the fragments with proteases to prepare for mass spectral analysis 780:858 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 4 26 theme peptides 883:890 arg1 fragments 769:777 Notch ECD fragments 759:777 Notch ECD fragments 759:777 Here we describe the expression and purification of Notch ECD fragments, digestion of the fragments with proteases to prepare for mass spectral analysis, and identification of peptides modified with O-glycans using mass spectrometry. 25053492 2 27 theme extracellular 212:224 arg1 ECD 234:236 ECD 234:236 ECD 234:236 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 27 theme extracellular 212:224 arg1 domain 226:231 The extracellular domain 208:231 The extracellular domain (ECD) 208:237 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 1 28 theme posttranslational 125:141 arg1 modifications 143:155 posttranslational modifications 125:155 posttranslational modifications 125:155 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 3 29 theme individual 610:619 arg1 sites 621:625 individual sites 610:625 individual sites on Notch 610:634 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 1 30 theme modifications 143:155 arg1 types 116:120 multiple types 107:120 multiple types 107:120 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 2 31 dep O-glycosylation 300:314 arg1 O-glucose 327:335 O-glucose 327:335 O-glucose 327:335 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 31 dep O-glycosylation 300:314 arg1 O-GlcNAc 342:349 O-GlcNAc 342:349 O-GlcNAc 342:349 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 31 dep O-glycosylation 300:314 arg1 O-fucose 317:324 O-fucose 317:324 O-fucose 317:324 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 1 32 mod modified 95:102 arg3 types 116:120 multiple types 107:120 multiple types 107:120 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 1 32 mod modified 95:102 arg1 Notch 86:90 Notch 86:90 Notch 86:90 Notch is modified by multiple types of posttranslational modifications, most of which are known to affect Notch function. 25053492 0 33 theme mass 67:70 arg1 spectrometry 72:83 mass spectrometry 67:83 mass spectrometry 67:83 Analyzing the posttranslational modification status of Notch using mass spectrometry. 25053492 2 34 mod modified 242:249 arg3 N-glycosylation 256:270 N-glycosylation 256:270 N-glycosylation 256:270 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 34 mod modified 242:249 arg1 ECD 234:236 ECD 234:236 ECD 234:236 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 34 mod modified 242:249 arg1 domain 226:231 The extracellular domain 208:231 The extracellular domain (ECD) 208:237 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 2 34 mod modified 242:249 arg3 types 291:295 at least three types 276:295 at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc) 276:350 The extracellular domain (ECD) is modified with N-glycosylation and at least three types of O-glycosylation (O-fucose, O-glucose, and O-GlcNAc), while the intracellular domain is hydroxylated, phosphorylated, and ubiquitinated. 25053492 3 35 theme O-glycans 490:498 arg1 structure 460:468 structure 460:468 structure 460:468 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 25053492 3 35 theme O-glycans 490:498 arg1 function 474:481 function 474:481 function 474:481 In order to analyze the structure and function of the O-glycans decorating the ECD, we have developed semiquantitative mass spectral methods for identifying modifications at individual sites on Notch that are generally applicable to most posttranslational modifications. 26699903 0 0 theme bone 92:95 arg1 signalling 119:128 bone morphogenetic protein signalling 92:128 bone morphogenetic protein signalling 92:128 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 6 1 theme haematopoietic 1416:1429 arg1 scl 1454:1456 scl 1454:1456 scl 1454:1456 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 1 theme haematopoietic 1416:1429 arg1 markers 1445:1451 haematopoietic and erythroid markers 1416:1451 haematopoietic and erythroid markers (scl and gata1) 1416:1467 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 1 theme haematopoietic 1416:1429 arg1 gata1 1462:1466 gata1 1462:1466 gata1 1462:1466 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 1 2 gly glycoprotein 174:185 arg1 SCUBE1 131:136 SCUBE1 131:136 SCUBE1 (S1) 131:141 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 2 gly glycoprotein 174:185 arg1 glycoprotein 174:185 a secreted and membrane-bound glycoprotein 144:185 a secreted and membrane-bound glycoprotein 144:185 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 3 theme secreted 146:153 arg1 SCUBE1 131:136 SCUBE1 131:136 SCUBE1 (S1) 131:141 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 3 theme secreted 146:153 arg1 glycoprotein 174:185 a secreted and membrane-bound glycoprotein 144:185 a secreted and membrane-bound glycoprotein 144:185 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 4 theme signal 246:251 arg1 sequence 261:268 an N-terminal signal peptide sequence 232:268 an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 232:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 4 5 theme amino 950:954 arg1 segment 941:947 a polycationic segment 926:947 a polycationic segment (amino acids 501-550) 926:969 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 4 5 theme amino 950:954 arg1 acids 956:960 amino acids 501-550 950:968 amino acids 501-550 950:968 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 2 6 theme membrane-associated 611:629 arg1 form 631:634 its membrane-associated form 607:634 its membrane-associated form 607:634 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 1 7 theme N-linked 407:414 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 3 8 theme S1 839:840 arg1 N-glycosylation 820:834 N-glycosylation 820:834 N-glycosylation of S1 820:840 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 1 9 theme cysteine-rich 357:369 arg1 motifs 376:381 three cysteine-rich (CR) motifs 351:381 three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites 351:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 0 10 theme protein 111:117 arg1 signalling 119:128 bone morphogenetic protein signalling 92:128 bone morphogenetic protein signalling 92:128 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 2 11 theme thrombosis 553:562 arg1 participant 538:548 an active participant 528:548 an active participant of thrombosis 528:562 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 1 12 theme glycosylation 416:428 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 13 with repeats 322:328 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 14 link N-linked 407:414 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 0 15 theme morphogenetic 97:109 arg1 signalling 119:128 bone morphogenetic protein signalling 92:128 bone morphogenetic protein signalling 92:128 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 1 16 theme spacer 333:338 arg1 region 340:345 a spacer region 331:345 a spacer region 331:345 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 6 17 dep markers 1445:1451 arg1 scl 1454:1456 scl 1454:1456 scl 1454:1456 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 17 dep markers 1445:1451 arg1 markers 1445:1451 haematopoietic and erythroid markers 1416:1451 haematopoietic and erythroid markers (scl and gata1) 1416:1467 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 17 dep markers 1445:1451 arg1 gata1 1462:1466 gata1 1462:1466 gata1 1462:1466 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 5 18 theme CR 1217:1218 arg1 motif 1220:1224 the CR motif 1213:1224 the CR motif 1213:1224 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 4 19 theme polycationic 928:939 arg1 segment 941:947 a polycationic segment 926:947 a polycationic segment (amino acids 501-550) 926:969 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 4 19 theme polycationic 928:939 arg1 acids 956:960 amino acids 501-550 950:968 amino acids 501-550 950:968 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 1 20 theme membrane-bound 159:172 arg1 SCUBE1 131:136 SCUBE1 131:136 SCUBE1 (S1) 131:141 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 20 theme membrane-bound 159:172 arg1 glycoprotein 174:185 a secreted and membrane-bound glycoprotein 144:185 a secreted and membrane-bound glycoprotein 144:185 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 2 21 theme Soluble 475:481 arg1 S1 483:484 Soluble S1 475:484 Soluble S1 475:484 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 2 21 theme Soluble 475:481 arg1 biomarker 491:499 a biomarker 489:499 a biomarker of platelet activation 489:522 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 6 22 from expression 1402:1411 arg1 embryos 1489:1495 scube1-knockdown embryos 1472:1495 scube1-knockdown embryos 1472:1495 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 2 23 theme bone 646:649 arg1 BMP 674:676 BMP 674:676 BMP 674:676 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 2 23 theme bone 646:649 arg1 protein 665:671 a bone morphogenetic protein 644:671 a bone morphogenetic protein (BMP) co-receptor 644:689 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 3 24 theme membrane 768:775 arg1 tethering 777:785 the membrane tethering 764:785 the membrane tethering 764:785 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 1 25 with motifs 376:381 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 6 26 theme scube1-knockdown 1472:1487 arg1 embryos 1489:1495 scube1-knockdown embryos 1472:1495 scube1-knockdown embryos 1472:1495 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 2 27 theme EGF-like 581:588 arg1 repeats 590:596 its adhesive EGF-like repeats 568:596 its adhesive EGF-like repeats 568:596 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 3 28 gly N-glycosylation 820:834 arg1 S1 839:840 S1 839:840 S1 839:840 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 2 29 theme BMP 704:706 arg1 activity 715:722 BMP signal activity 704:722 BMP signal activity 704:722 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 4 30 theme spacer 978:983 arg1 region 985:990 the spacer region 974:990 the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans 974:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 5 31 theme F 1170:1170 arg1 treatment 1172:1180 peptide N-glycosidase F treatment 1148:1180 peptide N-glycosidase F treatment 1148:1180 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 2 32 theme adhesive 572:579 arg1 repeats 590:596 its adhesive EGF-like repeats 568:596 its adhesive EGF-like repeats 568:596 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 1 33 theme CUB 445:447 arg1 domain 449:454 one CUB domain 441:454 one CUB domain 441:454 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 4 34 theme anionic 1077:1083 arg1 proteoglycans 1101:1113 the anionic heparan sulfate proteoglycans 1073:1113 the anionic heparan sulfate proteoglycans 1073:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 0 35 theme membrane 37:44 arg1 tethering 46:54 N-glycan-mediated membrane tethering 19:54 N-glycan-mediated membrane tethering 19:54 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 1 36 with domain 449:454 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 2 37 theme activation 513:522 arg1 S1 483:484 Soluble S1 475:484 Soluble S1 475:484 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 2 37 theme activation 513:522 arg1 biomarker 491:499 a biomarker 489:499 a biomarker of platelet activation 489:522 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 1 38 from C-terminus 463:472 arg1 region 340:345 a spacer region 331:345 a spacer region 331:345 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 38 from C-terminus 463:472 arg1 repeats 322:328 nine epidermal growth factor (EGF)-like repeats 282:328 nine epidermal growth factor (EGF)-like repeats 282:328 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 38 from C-terminus 463:472 arg1 motifs 376:381 three cysteine-rich (CR) motifs 351:381 three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites 351:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 38 from C-terminus 463:472 arg1 domain 449:454 one CUB domain 441:454 one CUB domain 441:454 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 0 39 theme N-glycan-mediated 19:35 arg1 tethering 46:54 N-glycan-mediated membrane tethering 19:54 N-glycan-mediated membrane tethering 19:54 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 2 40 theme platelet 504:511 arg1 activation 513:522 platelet activation 504:522 platelet activation 504:522 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 1 41 contain has 188:190 arg2 structure 210:218 a modular protein structure 192:218 a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 192:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 41 contain has 188:190 arg1 S1 139:140 S1 139:140 S1 139:140 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 41 contain has 188:190 arg1 SCUBE1 131:136 SCUBE1 131:136 SCUBE1 (S1) 131:141 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 41 contain has 188:190 arg1 glycoprotein 174:185 a secreted and membrane-bound glycoprotein 144:185 a secreted and membrane-bound glycoprotein 144:185 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 4 42 theme heparan 1085:1091 arg1 proteoglycans 1101:1113 the anionic heparan sulfate proteoglycans 1073:1113 the anionic heparan sulfate proteoglycans 1073:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 4 43 theme sulfate 1093:1099 arg1 proteoglycans 1101:1113 the anionic heparan sulfate proteoglycans 1073:1113 the anionic heparan sulfate proteoglycans 1073:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 1 44 theme modular 194:200 arg1 structure 210:218 a modular protein structure 192:218 a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 192:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 4 45 theme mapping 898:904 arg1 analysis 906:913 molecular mapping analysis 888:913 molecular mapping analysis 888:913 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 4 46 theme molecular 888:896 arg1 analysis 906:913 molecular mapping analysis 888:913 molecular mapping analysis 888:913 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 3 47 theme biological 795:804 arg1 importance 806:815 the biological importance 791:815 the biological importance of N-glycosylation of S1 791:840 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 1 48 theme protein 202:208 arg1 structure 210:218 a modular protein structure 192:218 a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 192:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 5 49 theme surface 1289:1295 arg1 retention 1297:1305 lectin-mediated surface retention 1273:1305 lectin-mediated surface retention 1273:1305 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 7 50 theme novel 1510:1514 arg1 mechanisms 1516:1525 novel mechanisms 1510:1525 novel mechanisms in targeting S1 to the plasma membrane 1510:1564 We describe novel mechanisms in targeting S1 to the plasma membrane and demonstrate that N-glycans are required for S1 functions during primitive haematopoiesis in zebrafish. 26699903 1 51 theme peptide 253:259 arg1 sequence 261:268 an N-terminal signal peptide sequence 232:268 an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 232:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 3 52 theme responsible 748:758 arg1 mechanism 738:746 the mechanism 734:746 the mechanism responsible for the membrane tethering 734:785 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 6 53 theme N-glycan-deficient 1363:1380 arg1 scube1 1382:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 5 54 theme lectin-mediated 1273:1287 arg1 retention 1297:1305 lectin-mediated surface retention 1273:1305 lectin-mediated surface retention 1273:1305 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 6 55 theme zebrafish 1335:1343 arg1 scube1 1382:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 7 56 theme primitive 1634:1642 arg1 haematopoiesis 1644:1657 primitive haematopoiesis 1634:1657 primitive haematopoiesis in zebrafish 1634:1670 We describe novel mechanisms in targeting S1 to the plasma membrane and demonstrate that N-glycans are required for S1 functions during primitive haematopoiesis in zebrafish. 26699903 6 57 theme mRNA 1321:1324 arg1 Injection 1308:1316 Injection 1308:1316 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 1308:1387 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 0 58 theme SCUBE1 59:64 arg1 Electrostatics 0:13 Electrostatics 0:13 Electrostatics 0:13 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 0 58 theme SCUBE1 59:64 arg1 tethering 46:54 N-glycan-mediated membrane tethering 19:54 N-glycan-mediated membrane tethering 19:54 Electrostatics and N-glycan-mediated membrane tethering of SCUBE1 is critical for promoting bone morphogenetic protein signalling. 26699903 5 59 theme peptide 1148:1154 arg1 F 1170:1170 peptide N-glycosidase F 1148:1170 peptide N-glycosidase F treatment 1148:1180 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 2 60 theme active 531:536 arg1 participant 538:548 an active participant 528:548 an active participant of thrombosis 528:562 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 2 61 theme protein 665:671 arg1 co-receptor 679:689 a bone morphogenetic protein (BMP) co-receptor 644:689 a bone morphogenetic protein (BMP) co-receptor 644:689 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 4 62 with interactions 1046:1057 arg1 proteoglycans 1101:1113 the anionic heparan sulfate proteoglycans 1073:1113 the anionic heparan sulfate proteoglycans 1073:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 3 63 theme N-glycosylation 820:834 arg1 importance 806:815 the biological importance 791:815 the biological importance of N-glycosylation of S1 791:840 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 3 63 theme N-glycosylation 820:834 arg1 mechanism 738:746 the mechanism 734:746 the mechanism responsible for the membrane tethering 734:785 However, the mechanism responsible for the membrane tethering and the biological importance of N-glycosylation of S1 remain largely unknown. 26699903 5 64 theme N-glycosidase 1156:1168 arg1 F 1170:1170 peptide N-glycosidase F 1148:1170 peptide N-glycosidase F treatment 1148:1180 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 4 65 theme present 873:879 arg1 study 881:885 the present study 869:885 the present study 869:885 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 7 66 theme plasma 1550:1555 arg1 membrane 1557:1564 the plasma membrane 1546:1564 the plasma membrane 1546:1564 We describe novel mechanisms in targeting S1 to the plasma membrane and demonstrate that N-glycans are required for S1 functions during primitive haematopoiesis in zebrafish. 26699903 2 67 theme morphogenetic 651:663 arg1 BMP 674:676 BMP 674:676 BMP 674:676 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 2 67 theme morphogenetic 651:663 arg1 protein 665:671 a bone morphogenetic protein 644:671 a bone morphogenetic protein (BMP) co-receptor 644:689 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 6 68 theme wild-type 1345:1353 arg1 scube1 1382:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 zebrafish wild-type but not N-glycan-deficient scube1 1335:1387 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 4 69 theme electrostatic 1032:1044 arg1 interactions 1046:1057 electrostatic interactions 1032:1057 electrostatic interactions possibly with the anionic heparan sulfate proteoglycans 1032:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 1 70 gly glycosylation 416:428 arg2 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 71 theme multiple 388:395 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 7 72 from haematopoiesis 1644:1657 arg1 zebrafish 1662:1670 zebrafish 1662:1670 zebrafish 1662:1670 We describe novel mechanisms in targeting S1 to the plasma membrane and demonstrate that N-glycans are required for S1 functions during primitive haematopoiesis in zebrafish. 26699903 2 73 theme signal 708:713 arg1 activity 715:722 BMP signal activity 704:722 BMP signal activity 704:722 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 6 74 theme erythroid 1435:1443 arg1 scl 1454:1456 scl 1454:1456 scl 1454:1456 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 74 theme erythroid 1435:1443 arg1 markers 1445:1451 haematopoietic and erythroid markers 1416:1451 haematopoietic and erythroid markers (scl and gata1) 1416:1467 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 6 74 theme erythroid 1435:1443 arg1 gata1 1462:1466 gata1 1462:1466 gata1 1462:1466 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 1 75 theme potential 397:405 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 2 76 dep biomarker 491:499 arg1 whereas 599:605 whereas 599:605 whereas 599:605 Soluble S1 is a biomarker of platelet activation but an active participant of thrombosis via its adhesive EGF-like repeats, whereas its membrane-associated form acts as a bone morphogenetic protein (BMP) co-receptor in promoting BMP signal activity. 26699903 5 77 theme membrane 1244:1251 arg1 recruitment 1253:1263 membrane recruitment 1244:1263 membrane recruitment through lectin-mediated surface retention 1244:1305 Furthermore, deglycosylation by peptide N-glycosidase F treatment revealed that N-glycans within the CR motif are essential for membrane recruitment through lectin-mediated surface retention. 26699903 6 78 theme markers 1445:1451 arg1 expression 1402:1411 the expression 1398:1411 the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos 1398:1495 Injection of mRNA encoding zebrafish wild-type but not N-glycan-deficient scube1 restores the expression of haematopoietic and erythroid markers (scl and gata1) in scube1-knockdown embryos. 26699903 1 79 with region 340:345 arg1 sites 430:434 multiple potential N-linked glycosylation sites 388:434 multiple potential N-linked glycosylation sites 388:434 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 80 dep cysteine-rich 357:369 arg1 CR 372:373 CR 372:373 CR 372:373 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 1 81 theme -like 316:320 arg1 repeats 322:328 nine epidermal growth factor (EGF)-like repeats 282:328 nine epidermal growth factor (EGF)-like repeats 282:328 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 26699903 4 82 theme membrane 1009:1016 arg1 tethering 1018:1026 its membrane tethering 1005:1026 its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans 1005:1113 In the present study, molecular mapping analysis identified a polycationic segment (amino acids 501-550) in the spacer region required for its membrane tethering via electrostatic interactions possibly with the anionic heparan sulfate proteoglycans. 26699903 1 83 theme N-terminal 235:244 arg1 sequence 261:268 an N-terminal signal peptide sequence 232:268 an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus 232:472 SCUBE1 (S1), a secreted and membrane-bound glycoprotein, has a modular protein structure composed of an N-terminal signal peptide sequence followed by nine epidermal growth factor (EGF)-like repeats, a spacer region and three cysteine-rich (CR) motifs with multiple potential N-linked glycosylation sites, and one CUB domain at the C-terminus. 28539451 11 0 theme active 2051:2056 arg1 antibodies 2058:2067 more broadly active antibodies 2038:2067 more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses 2038:2147 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 12 1 theme design 2184:2189 arg1 field 2191:2195 the trimer immunogen design field 2163:2195 the trimer immunogen design field 2163:2195 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 4 2 theme good 667:670 arg1 arguments 672:680 good arguments 667:680 good arguments in favor of suppressing such "off-target" immune responses 667:739 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 6 3 theme epitopes 1096:1103 arg1 presentation 1063:1074 the presentation 1059:1074 the presentation of trimer apex bNAb epitopes 1059:1103 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 13 4 theme off-target 2454:2463 arg1 antibodies 2465:2474 off-target antibodies 2454:2474 off-target antibodies 2454:2474 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 5 5 theme V3 804:805 arg1 epitopes 815:822 V3 non-NAb epitopes 804:822 V3 non-NAb epitopes 804:822 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 6 6 theme apex 1086:1089 arg1 epitopes 1096:1103 trimer apex bNAb epitopes 1079:1103 trimer apex bNAb epitopes 1079:1103 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 10 7 theme neutralizing 1711:1722 arg1 NAbs 1736:1739 NAbs 1736:1739 NAbs 1736:1739 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 7 theme neutralizing 1711:1722 arg1 antibodies 1724:1733 broadly active neutralizing antibodies 1696:1733 broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains 1696:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 1 8 theme immunodeficiency 176:191 arg1 type 199:202 human immunodeficiency virus type 1 170:204 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 8 theme immunodeficiency 176:191 arg1 HIV-1 207:211 HIV-1 207:211 HIV-1 207:211 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 8 9 theme additional 1185:1194 arg1 introduction 1196:1207 The additional introduction 1181:1207 The additional introduction of the E64K ground-state stabilizing substitution 1181:1257 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 2 10 theme multiple 283:290 arg1 bNAbs 325:329 bNAbs 325:329 bNAbs 325:329 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 2 10 theme multiple 283:290 arg1 antibodies 313:322 multiple broadly neutralizing antibodies 283:322 multiple broadly neutralizing antibodies (bNAbs) 283:330 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 1 11 theme type 199:202 arg1 programs 234:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 8 12 theme coreceptor 1369:1378 arg1 site 1388:1391 the coreceptor binding site 1365:1391 the coreceptor binding site 1365:1391 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 10 13 theme NAb 1550:1552 arg1 developed 1616:1624 developed 1616:1624 are being developed for future HIV-1 vaccine trials in humans 1606:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 13 theme NAb 1550:1552 arg1 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 13 theme NAb 1550:1552 arg1 response 1554:1561 the autologous NAb response 1535:1561 the autologous NAb response 1535:1561 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 8 14 theme E64K 1216:1219 arg1 substitution 1246:1257 the E64K ground-state stabilizing substitution 1212:1257 the E64K ground-state stabilizing substitution 1212:1257 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 13 15 theme V3 2417:2418 arg1 region 2420:2425 the gp120 V3 region 2407:2425 the gp120 V3 region 2407:2425 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 13 15 theme V3 2417:2418 arg1 site 2385:2388 the dominant site 2372:2388 the dominant site for tier-1 NAbs 2372:2404 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 9 16 theme E64K-modified 1416:1428 arg1 variant 1437:1443 E64K-modified trimer variant 1416:1443 E64K-modified trimer variant 1416:1443 When a V3 glycan- and E64K-modified trimer variant, BG505 SOSIP.664-E64K. 28539451 4 17 theme NAbs 647:650 arg1 induction 634:642 the induction 630:642 the induction of NAbs 630:650 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 8 18 theme stabilizing 1234:1244 arg1 substitution 1246:1257 the E64K ground-state stabilizing substitution 1212:1257 the E64K ground-state stabilizing substitution 1212:1257 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 1 19 theme Native-like 93:103 arg1 immunogens 156:165 immunogens 156:165 immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs 156:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 19 theme Native-like 93:103 arg1 trimers 105:111 Native-like trimers 93:111 Native-like trimers of the SOSIP design 93:131 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 11 20 theme narrow-specificity 1934:1951 arg1 antibodies 1953:1962 narrow-specificity antibodies 1934:1962 narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus 1934:2032 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 1 21 theme design 126:131 arg1 immunogens 156:165 immunogens 156:165 immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs 156:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 21 theme design 126:131 arg1 trimers 105:111 Native-like trimers 93:111 Native-like trimers of the SOSIP design 93:131 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 22 theme SOSIP 120:124 arg1 design 126:131 the SOSIP design 116:131 the SOSIP design 116:131 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 12 23 theme desired 2294:2300 arg1 responses 2302:2310 the more desired responses 2285:2310 the more desired responses to tier-2 epitopes 2285:2329 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 11 24 theme BG505 1899:1903 arg1 construct 1915:1923 the BG505 SOSIP.664 construct 1895:1923 the BG505 SOSIP.664 construct 1895:1923 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 1 25 from immunogens 156:165 arg1 programs 234:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 6 26 theme inserting 948:956 arg1 glycans 958:964 inserting glycans 948:964 inserting glycans at positions 306 and 314 (termed M1 and M7) 948:1008 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 0 27 theme Env 74:76 arg1 Trimers 84:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 6 28 from positions 969:977 arg1 glycans 958:964 inserting glycans 948:964 inserting glycans at positions 306 and 314 (termed M1 and M7) 948:1008 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 8 29 theme soluble 1287:1293 arg1 CD4 1295:1297 soluble CD4 1287:1297 soluble CD4 (sCD4) induction of non-NAb epitopes in V3 1287:1340 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 8 29 theme soluble 1287:1293 arg1 sCD4 1300:1303 sCD4 1300:1303 sCD4 1300:1303 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 5 30 theme BG505 913:917 arg1 trimers 925:931 BG505 SOSIP trimers 913:931 BG505 SOSIP trimers 913:931 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 12 31 theme off-target 2225:2234 arg1 antibodies 2236:2245 the latter off-target antibodies 2214:2245 the latter off-target antibodies 2214:2245 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 11 32 theme trimer 1861:1866 arg1 immunogens 1868:1877 native-like trimer immunogens 1849:1877 native-like trimer immunogens 1849:1877 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 4 33 theme immune 724:729 arg1 responses 731:739 such "off-target" immune responses 706:739 such "off-target" immune responses 706:739 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 5 34 theme trimers 925:931 arg1 region 903:908 the V3 region 896:908 the V3 region of BG505 SOSIP trimers 896:931 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 9 35 dep SOSIP.664-E64K 1452:1465 arg1 variant 1437:1443 E64K-modified trimer variant 1416:1443 E64K-modified trimer variant 1416:1443 When a V3 glycan- and E64K-modified trimer variant, BG505 SOSIP.664-E64K. 28539451 9 35 dep SOSIP.664-E64K 1452:1465 arg1 glycan- 1404:1410 a V3 glycan- 1399:1410 a V3 glycan- 1399:1410 When a V3 glycan- and E64K-modified trimer variant, BG505 SOSIP.664-E64K. 28539451 13 36 theme dominant 2376:2383 arg1 site 2385:2388 the dominant site 2372:2388 the dominant site for tier-1 NAbs 2372:2404 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 13 36 theme dominant 2376:2383 arg1 region 2420:2425 the gp120 V3 region 2407:2425 the gp120 V3 region 2407:2425 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 4 37 theme such 706:709 arg1 responses 731:739 such "off-target" immune responses 706:739 such "off-target" immune responses 706:739 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 11 38 theme native-like 1849:1859 arg1 immunogens 1868:1877 native-like trimer immunogens 1849:1877 native-like trimer immunogens 1849:1877 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 14 39 theme new 2498:2500 arg1 trimers 2502:2508 the new trimers 2494:2508 the new trimers 2494:2508 We characterized the new trimers, tested them as immunogens in rabbits, and found that the blocking glycans eliminated the induction of tier-1 NAbs to V3-epitopes. 28539451 8 40 theme epitopes 1327:1334 arg1 induction 1306:1314 soluble CD4 (sCD4) induction 1287:1314 soluble CD4 (sCD4) induction of non-NAb epitopes in V3 1287:1340 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 2 41 theme nonneutralizing 384:398 arg1 non-NAbs 412:419 non-NAbs 412:419 non-NAbs 412:419 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 2 41 theme nonneutralizing 384:398 arg1 antibodies 400:409 nonneutralizing antibodies 384:409 nonneutralizing antibodies (non-NAbs) 384:420 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 0 42 dep Soluble 47:53 arg1 Native-Like 56:66 Native-Like 56:66 Native-Like 56:66 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 10 43 theme wide 1768:1771 arg1 variety 1773:1779 a wide variety 1766:1779 a wide variety of circulating strains 1766:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 43 theme wide 1768:1771 arg1 strains 1796:1802 circulating strains 1784:1802 circulating strains 1784:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 12 44 theme trimer 2167:2172 arg1 field 2191:2195 the trimer immunogen design field 2163:2195 the trimer immunogen design field 2163:2195 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 3 45 theme gp120 460:464 arg1 region 469:474 the gp120 V3 region 456:474 the gp120 V3 region 456:474 Among the latter are epitopes in the gp120 V3 region that are highly immunogenic when SOSIP trimers are evaluated in animal models. 28539451 0 46 theme Soluble 47:53 arg1 Trimers 84:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 10 47 dep unchanged.IMPORTANCE 1567:1586 arg1 developed 1616:1624 developed 1616:1624 are being developed for future HIV-1 vaccine trials in humans 1606:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 47 dep unchanged.IMPORTANCE 1567:1586 arg1 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 47 dep unchanged.IMPORTANCE 1567:1586 arg1 response 1554:1561 the autologous NAb response 1535:1561 the autologous NAb response 1535:1561 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 3 48 from epitopes 444:451 arg1 region 469:474 the gp120 V3 region 456:474 the gp120 V3 region 456:474 Among the latter are epitopes in the gp120 V3 region that are highly immunogenic when SOSIP trimers are evaluated in animal models. 28539451 10 49 theme HIV-1 1637:1641 arg1 trials 1651:1656 future HIV-1 vaccine trials 1630:1656 future HIV-1 vaccine trials in humans 1630:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 4 50 from arguments 672:680 arg1 favor 685:689 favor 685:689 favor 685:689 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 8 51 theme substitution 1246:1257 arg1 introduction 1196:1207 The additional introduction 1181:1207 The additional introduction of the E64K ground-state stabilizing substitution 1181:1257 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 14 52 from immunogens 2526:2535 arg1 rabbits 2540:2546 rabbits 2540:2546 rabbits 2540:2546 We characterized the new trimers, tested them as immunogens in rabbits, and found that the blocking glycans eliminated the induction of tier-1 NAbs to V3-epitopes. 28539451 10 53 theme strains 1796:1802 arg1 variety 1773:1779 a wide variety 1766:1779 a wide variety of circulating strains 1766:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 53 theme strains 1796:1802 arg1 strains 1796:1802 circulating strains 1784:1802 circulating strains 1784:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 2 54 theme binding 352:358 arg1 sites 360:364 binding sites 352:364 binding sites for some types of nonneutralizing antibodies (non-NAbs) 352:420 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 12 55 from concern 2152:2158 arg1 field 2191:2195 the trimer immunogen design field 2163:2195 the trimer immunogen design field 2163:2195 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 1 56 theme vaccine 214:220 arg1 programs 234:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 10 57 with unchanged.IMPORTANCE 1567:1586 arg1 goal 1678:1681 the goal 1674:1681 the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains 1674:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 8 58 from induction 1306:1314 arg1 V3 1339:1340 V3 1339:1340 V3 1339:1340 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 11 59 theme sequence-divergent 2077:2094 arg1 viruses 2141:2147 sequence-divergent atypically neutralization-sensitive (tier-1) viruses 2077:2147 sequence-divergent atypically neutralization-sensitive (tier-1) viruses 2077:2147 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 7 60 theme Man6GlcNAc2 1163:1173 arg1 form 1175:1178 the Man6GlcNAc2 form 1159:1178 the Man6GlcNAc2 form 1159:1178 Both added glycans were shown to be predominantly of the Man6GlcNAc2 form. 28539451 10 61 theme V3 1496:1497 arg1 immunogenicity 1499:1512 V3 immunogenicity 1496:1512 V3 immunogenicity 1496:1512 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 3 62 theme SOSIP 509:513 arg1 trimers 515:521 SOSIP trimers 509:521 SOSIP trimers 509:521 Among the latter are epitopes in the gp120 V3 region that are highly immunogenic when SOSIP trimers are evaluated in animal models. 28539451 10 63 from trials 1651:1656 arg1 humans 1661:1666 humans 1661:1666 humans 1661:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 1 64 theme human 170:174 arg1 type 199:202 human immunodeficiency virus type 1 170:204 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 64 theme human 170:174 arg1 HIV-1 207:211 HIV-1 207:211 HIV-1 207:211 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 6 65 theme bNAb 1091:1094 arg1 epitopes 1096:1103 trimer apex bNAb epitopes 1079:1103 trimer apex bNAb epitopes 1079:1103 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 13 66 theme antibodies 2465:2474 arg1 induction 2441:2449 the induction 2437:2449 the induction of off-target antibodies 2437:2474 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 11 67 theme neutralization-resistant 1976:1999 arg1 virus 2028:2032 the neutralization-resistant (tier-2), sequence-matched virus 1972:2032 the neutralization-resistant (tier-2), sequence-matched virus 1972:2032 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 5 68 theme non-NAb 807:813 arg1 epitopes 815:822 V3 non-NAb epitopes 804:822 V3 non-NAb epitopes 804:822 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 11 69 theme neutralization-sensitive 2107:2130 arg1 viruses 2141:2147 sequence-divergent atypically neutralization-sensitive (tier-1) viruses 2077:2147 sequence-divergent atypically neutralization-sensitive (tier-1) viruses 2077:2147 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 6 70 theme trimer 1079:1084 arg1 epitopes 1096:1103 trimer apex bNAb epitopes 1079:1103 trimer apex bNAb epitopes 1079:1103 We found that inserting glycans at positions 306 and 314 (termed M1 and M7) markedly reduced V3 antigenicity while improving the presentation of trimer apex bNAb epitopes. 28539451 2 71 theme broadly 292:298 arg1 bNAbs 325:329 bNAbs 325:329 bNAbs 325:329 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 2 71 theme broadly 292:298 arg1 antibodies 313:322 multiple broadly neutralizing antibodies 283:322 multiple broadly neutralizing antibodies (bNAbs) 283:330 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 1 72 theme virus 193:197 arg1 type 199:202 human immunodeficiency virus type 1 170:204 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 1 72 theme virus 193:197 arg1 HIV-1 207:211 HIV-1 207:211 HIV-1 207:211 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28539451 2 73 theme neutralizing 300:311 arg1 bNAbs 325:329 bNAbs 325:329 bNAbs 325:329 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 2 73 theme neutralizing 300:311 arg1 antibodies 313:322 multiple broadly neutralizing antibodies 283:322 multiple broadly neutralizing antibodies (bNAbs) 283:330 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 14 74 theme blocking 2568:2575 arg1 glycans 2577:2583 the blocking glycans 2564:2583 the blocking glycans 2564:2583 We characterized the new trimers, tested them as immunogens in rabbits, and found that the blocking glycans eliminated the induction of tier-1 NAbs to V3-epitopes. 28539451 8 75 theme binding 1380:1386 arg1 site 1388:1391 the coreceptor binding site 1365:1391 the coreceptor binding site 1365:1391 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 10 76 theme vaccine 1643:1649 arg1 trials 1651:1656 future HIV-1 vaccine trials 1630:1656 future HIV-1 vaccine trials in humans 1630:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 5 77 theme epitopes 815:822 arg1 exposure 792:799 the exposure 788:799 the exposure of V3 non-NAb epitopes 788:822 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 9 78 theme V3 1401:1402 arg1 glycan- 1404:1410 a V3 glycan- 1399:1410 a V3 glycan- 1399:1410 When a V3 glycan- and E64K-modified trimer variant, BG505 SOSIP.664-E64K. 28539451 10 79 theme autologous 1539:1548 arg1 developed 1616:1624 developed 1616:1624 are being developed for future HIV-1 vaccine trials in humans 1606:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 79 theme autologous 1539:1548 arg1 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 unchanged.IMPORTANCE 1567:1586 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 79 theme autologous 1539:1548 arg1 response 1554:1561 the autologous NAb response 1535:1561 the autologous NAb response 1535:1561 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 13 80 theme gp120 2411:2415 arg1 region 2420:2425 the gp120 V3 region 2407:2425 the gp120 V3 region 2407:2425 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 13 80 theme gp120 2411:2415 arg1 site 2385:2388 the dominant site 2372:2388 the dominant site for tier-1 NAbs 2372:2404 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 11 81 theme immunogens 1868:1877 arg1 generation 1835:1844 the present generation 1823:1844 the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, 1823:1924 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 8 82 dep reduced 1268:1274 arg1 associated 1349:1358 associated 1349:1358 associated with the coreceptor binding site 1349:1391 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 8 82 dep reduced 1268:1274 arg1 ablated 1279:1285 ablated 1279:1285 ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 1279:1340 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 11 83 theme SOSIP.664 1905:1913 arg1 construct 1915:1923 the BG505 SOSIP.664 construct 1895:1923 the BG505 SOSIP.664 construct 1895:1923 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 9 84 theme trimer 1430:1435 arg1 variant 1437:1443 E64K-modified trimer variant 1416:1443 E64K-modified trimer variant 1416:1443 When a V3 glycan- and E64K-modified trimer variant, BG505 SOSIP.664-E64K. 28539451 10 85 theme active 1704:1709 arg1 NAbs 1736:1739 NAbs 1736:1739 NAbs 1736:1739 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 10 85 theme active 1704:1709 arg1 antibodies 1724:1733 broadly active neutralizing antibodies 1696:1733 broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains 1696:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 12 86 theme responses 2302:2310 arg1 induction 2272:2280 the induction 2268:2280 the induction of the more desired responses to tier-2 epitopes 2268:2329 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 8 87 theme ground-state 1221:1232 arg1 substitution 1246:1257 the E64K ground-state stabilizing substitution 1212:1257 the E64K ground-state stabilizing substitution 1212:1257 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 0 88 theme HIV-1 68:72 arg1 Trimers 84:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 3 89 theme animal 540:545 arg1 models 547:552 animal models 540:552 animal models 540:552 Among the latter are epitopes in the gp120 V3 region that are highly immunogenic when SOSIP trimers are evaluated in animal models. 28539451 0 90 theme SOSIP 78:82 arg1 Trimers 84:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Soluble, Native-Like HIV-1 Env SOSIP Trimers 47:90 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 11 91 dep neutralization-resistant 1976:1999 arg1 tier-2 2002:2007 tier-2 2002:2007 tier-2 2002:2007 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 11 91 dep neutralization-resistant 1976:1999 arg1 sequence-matched 2011:2026 sequence-matched 2011:2026 sequence-matched 2011:2026 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 12 92 theme tier-2 2315:2320 arg1 epitopes 2322:2329 tier-2 epitopes 2315:2329 tier-2 epitopes 2315:2329 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 13 93 theme tier-1 2394:2399 arg1 NAbs 2401:2404 tier-1 NAbs 2394:2404 tier-1 NAbs 2394:2404 Here, we have inserted two glycans into the dominant site for tier-1 NAbs, the gp120 V3 region, to block the induction of off-target antibodies. 28539451 14 94 theme NAbs 2620:2623 arg1 induction 2600:2608 the induction 2596:2608 the induction of tier-1 NAbs to V3-epitopes 2596:2638 We characterized the new trimers, tested them as immunogens in rabbits, and found that the blocking glycans eliminated the induction of tier-1 NAbs to V3-epitopes. 28539451 7 95 theme added 1111:1115 arg1 glycans 1117:1123 Both added glycans 1106:1123 Both added glycans 1106:1123 Both added glycans were shown to be predominantly of the Man6GlcNAc2 form. 28539451 5 96 theme SOSIP 919:923 arg1 trimers 925:931 BG505 SOSIP trimers 913:931 BG505 SOSIP trimers 913:931 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 8 97 theme CD4 1295:1297 arg1 induction 1306:1314 soluble CD4 (sCD4) induction 1287:1314 soluble CD4 (sCD4) induction of non-NAb epitopes in V3 1287:1340 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 10 98 theme Trimeric 1588:1595 arg1 proteins 1597:1604 Trimeric proteins 1588:1604 Trimeric proteins 1588:1604 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 0 99 theme V3 9:10 arg1 Antigenicity 12:23 V3 Antigenicity 9:23 V3 Antigenicity 9:23 Reducing V3 Antigenicity and Immunogenicity on Soluble, Native-Like HIV-1 Env SOSIP Trimers. 28539451 12 100 theme immunogen 2174:2182 arg1 field 2191:2195 the trimer immunogen design field 2163:2195 the trimer immunogen design field 2163:2195 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 5 101 theme V3 900:901 arg1 region 903:908 the V3 region 896:908 the V3 region of BG505 SOSIP trimers 896:931 Accordingly, we have assessed how to minimize the exposure of V3 non-NAb epitopes and thereby reduce their immunogenicity by introducing N-glycans within the V3 region of BG505 SOSIP trimers. 28539451 12 102 theme latter 2218:2223 arg1 antibodies 2236:2245 the latter off-target antibodies 2214:2245 the latter off-target antibodies 2214:2245 A concern in the trimer immunogen design field has been whether the latter off-target antibodies might interfere with the induction of the more desired responses to tier-2 epitopes. 28539451 10 103 theme circulating 1784:1794 arg1 strains 1796:1802 circulating strains 1784:1802 circulating strains 1784:1802 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 2 104 theme antibodies 400:409 arg1 types 375:379 some types 370:379 some types of nonneutralizing antibodies (non-NAbs) 370:420 These trimers display the epitopes for multiple broadly neutralizing antibodies (bNAbs) but can also expose binding sites for some types of nonneutralizing antibodies (non-NAbs). 28539451 11 105 dep neutralization-sensitive 2107:2130 arg1 tier-1 2133:2138 tier-1 2133:2138 tier-1 2133:2138 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 8 106 theme non-NAb 1319:1325 arg1 epitopes 1327:1334 non-NAb epitopes 1319:1334 non-NAb epitopes 1319:1334 The additional introduction of the E64K ground-state stabilizing substitution markedly reduced or ablated soluble CD4 (sCD4) induction of non-NAb epitopes in V3 and/or associated with the coreceptor binding site. 28539451 3 107 theme V3 466:467 arg1 region 469:474 the gp120 V3 region 456:474 the gp120 V3 region 456:474 Among the latter are epitopes in the gp120 V3 region that are highly immunogenic when SOSIP trimers are evaluated in animal models. 28539451 4 108 theme " 722:722 arg1 responses 731:739 such "off-target" immune responses 706:739 such "off-target" immune responses 706:739 It is presently uncertain whether antibodies against V3 can interfere with the induction of NAbs, but there are good arguments in favor of suppressing such "off-target" immune responses. 28539451 10 109 theme future 1630:1635 arg1 trials 1651:1656 future HIV-1 vaccine trials 1630:1656 future HIV-1 vaccine trials in humans 1630:1666 M1M7, was tested in rabbits, V3 immunogenicity was eliminated while the autologous NAb response was unchanged.IMPORTANCE Trimeric proteins are being developed for future HIV-1 vaccine trials in humans, with the goal of eliciting broadly active neutralizing antibodies (NAbs) that are active against a wide variety of circulating strains. 28539451 14 110 theme tier-1 2613:2618 arg1 NAbs 2620:2623 tier-1 NAbs 2613:2623 tier-1 NAbs 2613:2623 We characterized the new trimers, tested them as immunogens in rabbits, and found that the blocking glycans eliminated the induction of tier-1 NAbs to V3-epitopes. 28539451 11 111 theme present 1827:1833 arg1 generation 1835:1844 the present generation 1823:1844 the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, 1823:1924 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 11 112 theme animal 1808:1813 arg1 models 1815:1820 animal models 1808:1820 animal models 1808:1820 In animal models, the present generation of native-like trimer immunogens, exemplified by the BG505 SOSIP.664 construct, induces narrow-specificity antibodies against the neutralization-resistant (tier-2), sequence-matched virus and more broadly active antibodies against sequence-divergent atypically neutralization-sensitive (tier-1) viruses. 28539451 1 113 theme development 222:232 arg1 programs 234:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 human immunodeficiency virus type 1 (HIV-1) vaccine development programs 170:241 Native-like trimers of the SOSIP design are being developed as immunogens in human immunodeficiency virus type 1 (HIV-1) vaccine development programs. 28902916 11 0 from viruses 1865:1871 arg1 manner 1904:1909 a statistically significant manner 1876:1909 a statistically significant manner 1876:1909 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 7 1 theme trimer 1223:1228 arg1 variants 1230:1237 the N-glycan-deleted trimer variants 1202:1237 the N-glycan-deleted trimer variants 1202:1237 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 10 2 theme PNGS 1558:1561 arg1 sites 1563:1567 4 PNGS sites 1556:1567 4 PNGS sites 1556:1567 The first was to delete 4 PNGS sites and then boost with fully glycosylated Env; the second was to delete 4 sites and gradually re-introduce these N-glycans in subsequent boosts. 28902916 1 3 theme cell 241:244 arg1 recognition 246:256 B cell recognition 239:256 B cell recognition of conserved neutralizing determinants 239:295 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 12 4 theme viruses 2084:2090 arg1 capable 2038:2044 capable 2038:2044 capable 2038:2044 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 9 5 theme rabbit 1417:1422 arg1 experiments 1439:1449 rabbit immunogenicity experiments 1417:1449 rabbit immunogenicity experiments 1417:1449 We performed rabbit immunogenicity experiments using two approaches comparing glycan-deleted to fully glycosylated NFL trimers. 28902916 3 6 theme targeted 530:537 arg1 deletion 548:555 targeted N-glycan deletion 530:555 targeted N-glycan deletion 530:555 Accordingly, we sought to determine if targeted N-glycan deletion might alter antibody responses to Env. 28902916 12 7 theme cross-neutralizing 2049:2066 arg1 viruses 2084:2090 cross-neutralizing selected tier 2 viruses 2049:2090 cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered) 2049:2148 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 2 8 theme broadly 313:319 arg1 bNAbs 346:350 bNAbs 346:350 bNAbs 346:350 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 2 8 theme broadly 313:319 arg1 antibodies 334:343 broadly neutralizing antibodies 313:343 broadly neutralizing antibodies (bNAbs) 313:351 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 9 9 theme glycan-deleted 1482:1495 arg1 trimers 1523:1529 glycan-deleted to fully glycosylated NFL trimers 1482:1529 glycan-deleted to fully glycosylated NFL trimers 1482:1529 We performed rabbit immunogenicity experiments using two approaches comparing glycan-deleted to fully glycosylated NFL trimers. 28902916 12 10 theme tier 2077:2080 arg1 viruses 2084:2090 cross-neutralizing selected tier 2 viruses 2049:2090 cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered) 2049:2148 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 4 11 theme binding 628:634 arg1 site 636:639 the conserved CD4 binding site 610:639 the conserved CD4 binding site (CD4bs) 610:647 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 11 theme binding 628:634 arg1 CD4bs 642:646 CD4bs 642:646 CD4bs 642:646 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 0 12 theme elicited 113:120 arg1 response 131:138 the elicited antibody response 109:138 the elicited antibody response 109:138 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 1 13 theme HIV 196:198 arg1 Env 224:226 Env 224:226 Env 224:226 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 1 13 theme HIV 196:198 arg1 glycoproteins 209:221 the HIV envelope glycoproteins 192:221 the HIV envelope glycoproteins (Env) 192:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 12 14 theme serum 2028:2032 arg1 IgG 2034:2036 serum IgG 2028:2036 serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered) 2028:2148 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 7 15 from accessibility 1172:1184 arg1 variants 1230:1237 the N-glycan-deleted trimer variants 1202:1237 the N-glycan-deleted trimer variants 1202:1237 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 12 16 theme trimers 2197:2203 arg1 deletion 2172:2179 PNGS deletion 2167:2179 PNGS deletion of well-ordered trimers 2167:2203 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 12 16 theme trimers 2197:2203 arg1 strategy 2220:2227 a promising strategy 2208:2227 a promising strategy to prime B cell responses to this conserved neutralizing determinant 2208:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 9 17 theme immunogenicity 1424:1437 arg1 experiments 1439:1449 rabbit immunogenicity experiments 1417:1449 rabbit immunogenicity experiments 1417:1449 We performed rabbit immunogenicity experiments using two approaches comparing glycan-deleted to fully glycosylated NFL trimers. 28902916 1 18 theme glycoproteins 209:221 arg1 surface 181:187 the surface 177:187 the surface of the HIV envelope glycoproteins (Env) 177:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 12 19 theme capable 2038:2044 arg1 IgG 2034:2036 serum IgG 2028:2036 serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered) 2028:2148 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 10 20 gly glycosylated 1595:1606 arg1 Env 1608:1610 fully glycosylated Env 1589:1610 fully glycosylated Env 1589:1610 The first was to delete 4 PNGS sites and then boost with fully glycosylated Env; the second was to delete 4 sites and gradually re-introduce these N-glycans in subsequent boosts. 28902916 8 21 theme Env 1283:1285 arg1 PNGSs 1287:1291 these Env PNGSs 1277:1291 these Env PNGSs 1277:1291 We showed that pseudoviruses lacking these Env PNGSs were more sensitive to neutralization by CD4bs-specific bNAbs but remained resistant to non-neutralizing mAbs. 28902916 5 22 theme well-ordered 882:893 arg1 16055 903:907 well-ordered clade C 16055 native 882:914 well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 882:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 12 23 theme PNGS 2167:2170 arg1 deletion 2172:2179 PNGS deletion 2167:2179 PNGS deletion of well-ordered trimers 2167:2203 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 12 23 theme PNGS 2167:2170 arg1 strategy 2220:2227 a promising strategy 2208:2227 a promising strategy to prime B cell responses to this conserved neutralizing determinant 2208:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 5 24 theme C 901:901 arg1 16055 903:907 well-ordered clade C 16055 native 882:914 well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 882:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 9 25 gly glycosylated 1506:1517 arg1 NFL 1519:1521 fully glycosylated NFL 1500:1521 fully glycosylated NFL 1500:1521 We performed rabbit immunogenicity experiments using two approaches comparing glycan-deleted to fully glycosylated NFL trimers. 28902916 11 26 theme glycosylated 1974:1985 arg1 virus 1998:2002 fully glycosylated autologous virus 1968:2002 fully glycosylated autologous virus 1968:2002 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 7 27 theme N-glycan-deleted 1206:1221 arg1 variants 1230:1237 the N-glycan-deleted trimer variants 1202:1237 the N-glycan-deleted trimer variants 1202:1237 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 4 28 theme neutralizing 671:682 arg1 this 655:658 this 655:658 this 655:658 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 28 theme neutralizing 671:682 arg1 determinant 684:694 a known neutralizing determinant 663:694 a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans 663:799 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 28 theme neutralizing 671:682 arg1 devoid 704:709 devoid 704:709 devoid 704:709 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 0 29 theme NFL 72:74 arg1 integrity 83:91 HIV Env NFL trimer integrity 64:91 HIV Env NFL trimer integrity 64:91 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 8 30 theme CD4bs-specific 1334:1347 arg1 bNAbs 1349:1353 CD4bs-specific bNAbs 1334:1353 CD4bs-specific bNAbs 1334:1353 We showed that pseudoviruses lacking these Env PNGSs were more sensitive to neutralization by CD4bs-specific bNAbs but remained resistant to non-neutralizing mAbs. 28902916 11 31 theme serum 1785:1789 arg1 antibodies 1791:1800 serum antibodies 1785:1800 serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner 1785:1909 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 5 32 theme NFL 933:935 arg1 trimers 938:944 (NFL) trimers 932:944 (NFL) trimers 932:944 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 0 33 from site 51:54 arg1 deletion 18:25 Targeted N-glycan deletion 0:25 Targeted N-glycan deletion at the receptor-binding site 0:54 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 2 34 theme capable 407:413 arg1 Abs 403:405 Abs 403:405 Abs capable of penetrating the glycan shield 403:446 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 6 35 theme trimer 1036:1041 arg1 variants 1043:1050 glycan-deleted trimer variants 1021:1050 glycan-deleted trimer variants that maintained native-like conformation and stability 1021:1105 We generated glycan-deleted trimer variants that maintained native-like conformation and stability. 28902916 4 36 theme glycosylation 714:726 arg1 this 655:658 this 655:658 this 655:658 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 36 theme glycosylation 714:726 arg1 determinant 684:694 a known neutralizing determinant 663:694 a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans 663:799 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 36 theme glycosylation 714:726 arg1 devoid 704:709 devoid 704:709 devoid 704:709 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 0 37 theme Targeted 0:7 arg1 deletion 18:25 Targeted N-glycan deletion 0:25 Targeted N-glycan deletion at the receptor-binding site 0:54 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 1 38 theme conserved 261:269 arg1 determinants 284:295 conserved neutralizing determinants 261:295 conserved neutralizing determinants 261:295 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 5 39 theme native 909:914 arg1 16055 903:907 well-ordered clade C 16055 native 882:914 well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 882:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 8 40 theme non-neutralizing 1381:1396 arg1 mAbs 1398:1401 non-neutralizing mAbs 1381:1401 non-neutralizing mAbs 1381:1401 We showed that pseudoviruses lacking these Env PNGSs were more sensitive to neutralization by CD4bs-specific bNAbs but remained resistant to non-neutralizing mAbs. 28902916 1 41 theme determinants 284:295 arg1 recognition 246:256 B cell recognition 239:256 B cell recognition of conserved neutralizing determinants 239:295 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 1 42 theme Extensive 141:149 arg1 shielding 151:159 Extensive shielding 141:159 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) 141:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 2 43 theme cell 474:477 arg1 repertoire 479:488 the B cell repertoire 468:488 the B cell repertoire 468:488 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 11 44 theme virus 1998:2002 arg1 neutralization 1950:1963 increased neutralization 1940:1963 increased neutralization of fully glycosylated autologous virus 1940:2002 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 7 45 theme CD4bs-directed 1125:1138 arg1 bNAbs 1140:1144 CD4bs-directed bNAbs 1125:1144 CD4bs-directed bNAbs 1125:1144 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 5 46 theme naïve 985:989 arg1 cells 993:997 naïve B cells 985:997 naïve B cells 985:997 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 0 47 theme HIV 64:66 arg1 integrity 83:91 HIV Env NFL trimer integrity 64:91 HIV Env NFL trimer integrity 64:91 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 2 48 dep glycan 434:439 arg1 shield 441:446 shield 441:446 shield 441:446 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 12 49 dep residue 2113:2119 arg1 engineered 2138:2147 engineered 2138:2147 engineered 2138:2147 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 12 49 dep residue 2113:2119 arg1 natural 2127:2133 natural 2127:2133 natural 2127:2133 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 11 50 theme PNGS-deleted 1742:1753 arg1 trimers 1755:1761 the 16055 PNGS-deleted trimers 1732:1761 the 16055 PNGS-deleted trimers 1732:1761 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 12 51 theme prime 2232:2236 arg1 responses 2245:2253 prime B cell responses 2232:2253 prime B cell responses to this conserved neutralizing determinant 2232:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 11 52 theme significant 1892:1902 arg1 manner 1904:1909 a statistically significant manner 1876:1909 a statistically significant manner 1876:1909 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 12 53 theme cell 2240:2243 arg1 responses 2245:2253 prime B cell responses 2232:2253 prime B cell responses to this conserved neutralizing determinant 2232:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 11 54 theme increased 1940:1948 arg1 neutralization 1950:1963 increased neutralization 1940:1963 increased neutralization of fully glycosylated autologous virus 1940:2002 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 6 55 theme native-like 1068:1078 arg1 conformation 1080:1091 native-like conformation 1068:1091 native-like conformation 1068:1091 We generated glycan-deleted trimer variants that maintained native-like conformation and stability. 28902916 7 56 theme CD4bs 1193:1197 arg1 accessibility 1172:1184 accessibility 1172:1184 accessibility of the CD4bs on the N-glycan-deleted trimer variants 1172:1237 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 2 57 theme selected 356:363 arg1 individuals 378:388 selected HIV-infected individuals 356:388 selected HIV-infected individuals 356:388 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 1 58 theme B 239:239 arg1 recognition 246:256 B cell recognition 239:256 B cell recognition of conserved neutralizing determinants 239:295 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 10 59 theme glycosylated 1595:1606 arg1 Env 1608:1610 fully glycosylated Env 1589:1610 fully glycosylated Env 1589:1610 The first was to delete 4 PNGS sites and then boost with fully glycosylated Env; the second was to delete 4 sites and gradually re-introduce these N-glycans in subsequent boosts. 28902916 10 60 dep subsequent 1692:1701 arg1 boosts 1703:1708 boosts 1703:1708 boosts 1703:1708 The first was to delete 4 PNGS sites and then boost with fully glycosylated Env; the second was to delete 4 sites and gradually re-introduce these N-glycans in subsequent boosts. 28902916 3 61 theme N-glycan 539:546 arg1 deletion 548:555 targeted N-glycan deletion 530:555 targeted N-glycan deletion 530:555 Accordingly, we sought to determine if targeted N-glycan deletion might alter antibody responses to Env. 28902916 2 62 theme neutralizing 321:332 arg1 bNAbs 346:350 bNAbs 346:350 bNAbs 346:350 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 2 62 theme neutralizing 321:332 arg1 antibodies 334:343 broadly neutralizing antibodies 313:343 broadly neutralizing antibodies (bNAbs) 313:351 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 5 63 from recognition 970:980 arg1 vivo 1002:1005 vivo 1002:1005 vivo 1002:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 1 64 gly glycoproteins 209:221 arg1 Env 224:226 Env 224:226 Env 224:226 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 1 64 gly glycoproteins 209:221 arg1 glycoproteins 209:221 the HIV envelope glycoproteins 192:221 the HIV envelope glycoproteins (Env) 192:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 3 65 theme antibody 569:576 arg1 responses 578:586 antibody responses 569:586 antibody responses to Env 569:593 Accordingly, we sought to determine if targeted N-glycan deletion might alter antibody responses to Env. 28902916 12 66 theme promising 2210:2218 arg1 deletion 2172:2179 PNGS deletion 2167:2179 PNGS deletion of well-ordered trimers 2167:2203 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 12 66 theme promising 2210:2218 arg1 strategy 2220:2227 a promising strategy 2208:2227 a promising strategy to prime B cell responses to this conserved neutralizing determinant 2208:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 11 67 theme CD4bs-proximal-PNGS-deleted 1837:1863 arg1 viruses 1865:1871 the CD4bs-proximal-PNGS-deleted viruses 1833:1871 the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner 1833:1909 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 0 68 theme antibody 122:129 arg1 response 131:138 the elicited antibody response 109:138 the elicited antibody response 109:138 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 12 69 theme selected 2068:2075 arg1 viruses 2084:2090 cross-neutralizing selected tier 2 viruses 2049:2090 cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered) 2049:2148 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 4 70 theme CD4 624:626 arg1 site 636:639 the conserved CD4 binding site 610:639 the conserved CD4 binding site (CD4bs) 610:647 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 70 theme CD4 624:626 arg1 CD4bs 642:646 CD4bs 642:646 CD4bs 642:646 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 9 71 theme glycosylated 1506:1517 arg1 NFL 1519:1521 fully glycosylated NFL 1500:1521 fully glycosylated NFL 1500:1521 We performed rabbit immunogenicity experiments using two approaches comparing glycan-deleted to fully glycosylated NFL trimers. 28902916 12 72 theme well-ordered 2184:2195 arg1 trimers 2197:2203 well-ordered trimers 2184:2203 well-ordered trimers 2184:2203 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 5 73 theme clade 895:899 arg1 16055 903:907 well-ordered clade C 16055 native 882:914 well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 882:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 4 74 theme receptor 741:748 arg1 engagement 750:759 CD4 receptor engagement 737:759 CD4 receptor engagement 737:759 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 0 75 theme Env 68:70 arg1 integrity 83:91 HIV Env NFL trimer integrity 64:91 HIV Env NFL trimer integrity 64:91 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 5 76 theme proximal 857:864 arg1 PNGS 851:854 PNGS 851:854 PNGS 851:854 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 5 76 theme proximal 857:864 arg1 sites 844:848 potential N-glycan sites 825:848 potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 825:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 0 77 theme trimer 76:81 arg1 integrity 83:91 HIV Env NFL trimer integrity 64:91 HIV Env NFL trimer integrity 64:91 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 2 78 from Elicitation 298:308 arg1 individuals 378:388 selected HIV-infected individuals 356:388 selected HIV-infected individuals 356:388 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 4 79 theme known 665:669 arg1 this 655:658 this 655:658 this 655:658 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 79 theme known 665:669 arg1 determinant 684:694 a known neutralizing determinant 663:694 a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans 663:799 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 79 theme known 665:669 arg1 devoid 704:709 devoid 704:709 devoid 704:709 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 5 80 from CD4bs 873:877 arg1 16055 903:907 well-ordered clade C 16055 native 882:914 well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 882:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 7 81 dep demonstrated 1150:1161 arg1 improved 1163:1170 improved 1163:1170 demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants 1150:1237 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 4 82 theme CD4 737:739 arg1 engagement 750:759 CD4 receptor engagement 737:759 CD4 receptor engagement 737:759 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 12 83 theme conserved 2263:2271 arg1 determinant 2286:2296 this conserved neutralizing determinant 2258:2296 this conserved neutralizing determinant 2258:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 1 84 from shielding 151:159 arg1 surface 181:187 the surface 177:187 the surface of the HIV envelope glycoproteins (Env) 177:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 1 85 theme envelope 200:207 arg1 Env 224:226 Env 224:226 Env 224:226 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 1 85 theme envelope 200:207 arg1 glycoproteins 209:221 the HIV envelope glycoproteins 192:221 the HIV envelope glycoproteins (Env) 192:227 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 11 86 theme autologous 1987:1996 arg1 virus 1998:2002 fully glycosylated autologous virus 1968:2002 fully glycosylated autologous virus 1968:2002 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 0 87 theme N-glycan 9:16 arg1 deletion 18:25 Targeted N-glycan deletion 0:25 Targeted N-glycan deletion at the receptor-binding site 0:54 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 11 88 gly glycosylated 1974:1985 arg1 virus 1998:2002 fully glycosylated autologous virus 1968:2002 fully glycosylated autologous virus 1968:2002 We demonstrated that the 16055 PNGS-deleted trimers more rapidly elicited serum antibodies that more potently neutralized the CD4bs-proximal-PNGS-deleted viruses in a statistically significant manner and strongly trended towards increased neutralization of fully glycosylated autologous virus. 28902916 6 89 theme glycan-deleted 1021:1034 arg1 variants 1043:1050 glycan-deleted trimer variants 1021:1050 glycan-deleted trimer variants that maintained native-like conformation and stability 1021:1105 We generated glycan-deleted trimer variants that maintained native-like conformation and stability. 28902916 12 90 theme neutralizing 2273:2284 arg1 determinant 2286:2296 this conserved neutralizing determinant 2258:2296 this conserved neutralizing determinant 2258:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 7 91 theme bNAbs 1140:1144 arg1 panel 1116:1120 a panel 1114:1120 a panel of CD4bs-directed bNAbs 1114:1144 Using a panel of CD4bs-directed bNAbs, we demonstrated improved accessibility of the CD4bs on the N-glycan-deleted trimer variants. 28902916 1 92 theme neutralizing 271:282 arg1 determinants 284:295 conserved neutralizing determinants 261:295 conserved neutralizing determinants 261:295 Extensive shielding by N-glycans on the surface of the HIV envelope glycoproteins (Env) restricts B cell recognition of conserved neutralizing determinants. 28902916 0 93 theme receptor-binding 34:49 arg1 site 51:54 the receptor-binding site 30:54 the receptor-binding site 30:54 Targeted N-glycan deletion at the receptor-binding site retains HIV Env NFL trimer integrity and accelerates the elicited antibody response. 28902916 5 94 theme potential 825:833 arg1 PNGS 851:854 PNGS 851:854 PNGS 851:854 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 5 94 theme potential 825:833 arg1 sites 844:848 potential N-glycan sites 825:848 potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 825:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 2 95 theme B 472:472 arg1 repertoire 479:488 the B cell repertoire 468:488 the B cell repertoire 468:488 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 4 96 theme surrounding 779:789 arg1 N-glycans 791:799 surrounding N-glycans 779:799 surrounding N-glycans 779:799 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 5 97 theme B 991:991 arg1 cells 993:997 naïve B cells 985:997 naïve B cells 985:997 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 2 98 theme antibodies 334:343 arg1 Elicitation 298:308 Elicitation 298:308 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals 298:388 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 28902916 5 99 theme N-glycan 835:842 arg1 PNGS 851:854 PNGS 851:854 PNGS 851:854 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 5 99 theme N-glycan 835:842 arg1 sites 844:848 potential N-glycan sites 825:848 potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo 825:1005 We selectively deleted potential N-glycan sites (PNGS) proximal to the CD4bs on well-ordered clade C 16055 native flexibly linked (NFL) trimers to potentially increase recognition by naïve B cells in vivo. 28902916 4 100 theme conserved 614:622 arg1 site 636:639 the conserved CD4 binding site 610:639 the conserved CD4 binding site (CD4bs) 610:647 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 4 100 theme conserved 614:622 arg1 CD4bs 642:646 CD4bs 642:646 CD4bs 642:646 We focused on the conserved CD4 binding site (CD4bs) since this is a known neutralizing determinant that is devoid of glycosylation to allow CD4 receptor engagement, but is ringed by surrounding N-glycans. 28902916 12 101 theme B 2238:2238 arg1 responses 2245:2253 prime B cell responses 2232:2253 prime B cell responses to this conserved neutralizing determinant 2232:2296 This approach elicited serum IgG capable of cross-neutralizing selected tier 2 viruses lacking N-glycans at residue N276 (natural or engineered), indicating that PNGS deletion of well-ordered trimers is a promising strategy to prime B cell responses to this conserved neutralizing determinant. 28902916 2 102 theme HIV-infected 365:376 arg1 individuals 378:388 selected HIV-infected individuals 356:388 selected HIV-infected individuals 356:388 Elicitation of broadly neutralizing antibodies (bNAbs) in selected HIV-infected individuals reveals that Abs capable of penetrating the glycan shield can be generated by the B cell repertoire. 25422509 0 0 theme T 82:82 arg1 cells 84:88 T cells 82:88 T cells 82:88 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 4 1 located present 629:635 arg1 serum 644:648 the serum 640:648 the serum of healthy individuals 640:671 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 4 1 located present 629:635 arg2 IgM 622:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 1 2 theme glycosylated 135:146 arg1 polymers 148:155 heavily glycosylated polymers 127:155 heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain 127:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 7 3 theme key 1098:1100 arg1 role 1102:1105 a key role 1096:1105 a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression 1096:1177 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 1 4 with polymers 148:155 arg1 sites 181:185 five glycosylation sites 162:185 five glycosylation sites on the μ (heavy) chain 162:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 4 with polymers 148:155 arg1 site 232:235 one glycosylation site 214:235 one glycosylation site on the J chain 214:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 6 5 theme responses 1002:1010 arg1 inhibition 981:990 severe inhibition 974:990 severe inhibition of T cell responses 974:1010 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 1 6 theme glycosylation 218:230 arg1 site 232:235 one glycosylation site 214:235 one glycosylation site on the J chain 214:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 6 7 theme severe 974:979 arg1 inhibition 981:990 severe inhibition 974:990 severe inhibition of T cell responses 974:1010 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 0 8 link N-linked 15:22 arg1 glycans 24:30 N-linked glycans 15:30 N-linked glycans 15:30 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 4 9 theme inflammatory 690:701 arg1 conditions 703:712 inflammatory conditions 690:712 inflammatory conditions 690:712 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 6 10 link N-linked 890:897 arg1 glycans 899:905 sialylated N-linked glycans 879:905 sialylated N-linked glycans 879:905 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 6 11 theme sialylated 879:888 arg1 glycans 899:905 sialylated N-linked glycans 879:905 sialylated N-linked glycans 879:905 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 3 12 dep Ig 445:446 arg1 whereas 504:510 whereas 504:510 whereas 504:510 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 0 13 from effects 64:70 arg1 cells 84:88 T cells 82:88 T cells 82:88 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 1 14 gly glycosylation 167:179 arg2 sites 181:185 five glycosylation sites 162:185 five glycosylation sites on the μ (heavy) chain 162:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 14 gly glycosylation 167:179 arg2 five 162:165 five 162:165 five 162:165 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 7 15 theme N-glycans 1121:1129 arg1 role 1102:1105 a key role 1096:1105 a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression 1096:1177 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 3 16 theme low 490:492 arg1 affinity 494:501 low affinity 490:501 low affinity 490:501 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 6 17 theme T 995:995 arg1 responses 1002:1010 T cell responses 995:1010 T cell responses 995:1010 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 7 18 theme inhibitory 1065:1074 arg1 activities 1076:1085 these inhibitory activities 1059:1085 these inhibitory activities 1059:1085 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 4 19 theme healthy 653:659 arg1 individuals 661:671 healthy individuals 653:671 healthy individuals 653:671 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 7 20 gly sialylated 1110:1119 arg1 N-glycans 1121:1129 sialylated N-glycans 1110:1129 sialylated N-glycans 1110:1129 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 4 21 theme individuals 661:671 arg1 serum 644:648 the serum 640:648 the serum of healthy individuals 640:671 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 3 22 theme multiple 472:479 arg1 Ags 481:483 multiple Ags 472:483 multiple Ags with low affinity 472:501 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 2 23 theme biological 313:322 arg1 functions 324:332 biological functions 313:332 biological functions 313:332 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 5 24 theme molecular 775:783 arg1 mechanism 785:793 the underlying molecular mechanism 760:793 the underlying molecular mechanism 760:793 It is able to inhibit T cell activation, but the underlying molecular mechanism is not understood. 25422509 2 25 theme IgM 404:406 arg1 glycans 408:414 IgM glycans 404:414 IgM glycans 404:414 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 1 26 gly glycosylation 218:230 arg2 site 232:235 one glycosylation site 214:235 one glycosylation site on the J chain 214:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 26 gly glycosylation 218:230 arg2 one 214:216 one 214:216 one 214:216 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 2 27 theme functions 324:332 arg1 number 303:308 a number 301:308 a number of biological functions 301:332 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 4 28 attach present 629:635 arg1 serum 644:648 the serum 640:648 the serum of healthy individuals 640:671 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 4 28 attach present 629:635 arg2 IgM 622:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 2 29 theme IgG 268:270 arg1 glycans 272:278 IgG glycans 268:278 IgG glycans 268:278 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 7 30 theme IgM-mediated 1147:1158 arg1 suppression 1167:1177 the IgM-mediated immune suppression 1143:1177 the IgM-mediated immune suppression 1143:1177 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 1 31 theme glycosylation 167:179 arg1 sites 181:185 five glycosylation sites 162:185 five glycosylation sites on the μ (heavy) chain 162:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 5 32 theme T 737:737 arg1 activation 744:753 T cell activation 737:753 T cell activation 737:753 It is able to inhibit T cell activation, but the underlying molecular mechanism is not understood. 25422509 7 33 theme sialylated 1110:1119 arg1 N-glycans 1121:1129 sialylated N-glycans 1110:1129 sialylated N-glycans 1110:1129 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 2 34 dep glycans 272:278 arg1 contrast 256:263 contrast 256:263 contrast 256:263 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 0 35 theme glycans 24:30 arg1 Sialylation 0:10 Sialylation 0:10 Sialylation of N-linked glycans 0:30 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 1 36 theme J 244:244 arg1 chain 246:250 the J chain 240:250 the J chain 240:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 37 from site 232:235 arg1 chain 204:208 the μ (heavy) chain 190:208 the μ (heavy) chain 190:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 37 from site 232:235 arg1 chain 246:250 the J chain 240:250 the J chain 240:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 38 theme Human 91:95 arg1 Abs 107:109 Human serum IgM Abs 91:109 Human serum IgM Abs 91:109 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 0 39 theme N-linked 15:22 arg1 glycans 24:30 N-linked glycans 15:30 N-linked glycans 15:30 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 1 40 theme serum 97:101 arg1 Abs 107:109 Human serum IgM Abs 91:109 Human serum IgM Abs 91:109 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 7 41 theme sialic 1028:1033 arg1 residues 1040:1047 sialic acid residues 1028:1047 sialic acid residues 1028:1047 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 3 42 theme Natural 417:423 arg1 IgM 425:427 Natural IgM 417:427 Natural IgM 417:427 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 3 42 theme Natural 417:423 arg1 Ig 445:446 the earliest Ig 432:446 the earliest Ig produced 432:455 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 4 43 theme Natural 598:604 arg1 IgM 622:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 3 44 theme higher 575:580 arg1 specificity 585:595 a higher Ag specificity 573:595 a higher Ag specificity 573:595 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 1 45 theme μ 194:194 arg1 chain 204:208 the μ (heavy) chain 190:208 the μ (heavy) chain 190:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 3 46 with Ags 481:483 arg1 affinity 494:501 low affinity 490:501 low affinity 490:501 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 6 47 theme N-linked 890:897 arg1 glycans 899:905 sialylated N-linked glycans 879:905 sialylated N-linked glycans 879:905 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 2 48 theme structure-function 368:385 arg1 relationships 387:399 structure-function relationships 368:399 structure-function relationships of IgM glycans 368:414 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 6 49 theme T 944:944 arg1 cells 946:950 T cells 944:950 T cells 944:950 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 5 50 theme cell 739:742 arg1 activation 744:753 T cell activation 737:753 T cell activation 737:753 It is able to inhibit T cell activation, but the underlying molecular mechanism is not understood. 25422509 2 51 theme glycans 408:414 arg1 relationships 387:399 structure-function relationships 368:399 structure-function relationships of IgM glycans 368:414 In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. 25422509 6 52 theme IgM 937:939 arg1 internalization 918:932 the internalization 914:932 the internalization of IgM 914:939 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 6 53 theme first 863:867 arg1 time 869:872 the first time 859:872 the first time 859:872 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 1 54 theme IgM 103:105 arg1 Abs 107:109 Human serum IgM Abs 91:109 Human serum IgM Abs 91:109 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 0 55 theme immunomodulatory 47:62 arg1 effects 64:70 the immunomodulatory effects 43:70 the immunomodulatory effects of IgM on T cells 43:88 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 7 56 theme immune 1160:1165 arg1 suppression 1167:1177 the IgM-mediated immune suppression 1143:1177 the IgM-mediated immune suppression 1143:1177 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 3 57 theme earliest 436:443 arg1 IgM 425:427 Natural IgM 417:427 Natural IgM 417:427 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 3 57 theme earliest 436:443 arg1 Ig 445:446 the earliest Ig 432:446 the earliest Ig produced 432:455 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 7 58 theme acid 1035:1038 arg1 residues 1040:1047 sialic acid residues 1028:1047 sialic acid residues 1028:1047 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 6 59 gly sialylated 879:888 arg1 glycans 899:905 sialylated N-linked glycans 879:905 sialylated N-linked glycans 879:905 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 6 60 theme cell 997:1000 arg1 responses 1002:1010 T cell responses 995:1010 T cell responses 995:1010 In this study, to our knowledge, we show for the first time that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. 25422509 4 61 from serum 644:648 arg1 present 629:635 present 629:635 present 629:635 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 1 62 theme heavy 197:201 arg1 chain 204:208 the μ (heavy) chain 190:208 the μ (heavy) chain 190:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 63 gly glycosylated 135:146 arg1 polymers 148:155 heavily glycosylated polymers 127:155 heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain 127:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 3 64 theme Ag 582:583 arg1 specificity 585:595 a higher Ag specificity 573:595 a higher Ag specificity 573:595 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 4 65 theme anti-lymphocyte 606:620 arg1 IgM 622:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM 598:624 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25422509 0 66 theme IgM 75:77 arg1 effects 64:70 the immunomodulatory effects 43:70 the immunomodulatory effects of IgM on T cells 43:88 Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells. 25422509 5 67 theme underlying 764:773 arg1 mechanism 785:793 the underlying molecular mechanism 760:793 the underlying molecular mechanism 760:793 It is able to inhibit T cell activation, but the underlying molecular mechanism is not understood. 25422509 3 68 theme Ag 537:538 arg1 exposure 540:547 Ag exposure 537:547 Ag exposure 537:547 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 1 69 from sites 181:185 arg1 chain 204:208 the μ (heavy) chain 190:208 the μ (heavy) chain 190:208 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 1 69 from sites 181:185 arg1 chain 246:250 the J chain 240:250 the J chain 240:250 Human serum IgM Abs are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. 25422509 7 70 theme residues 1040:1047 arg1 absence 1017:1023 The absence 1013:1023 The absence of sialic acid residues 1013:1047 The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression. 25422509 3 71 theme immune 512:517 arg1 IgM 519:521 immune IgM 512:521 immune IgM 512:521 Natural IgM is the earliest Ig produced and recognizes multiple Ags with low affinity, whereas immune IgM is induced by Ag exposure and is characterized by a higher Ag specificity. 25422509 4 72 from present 629:635 arg1 serum 644:648 the serum 640:648 the serum of healthy individuals 640:671 Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. 25452312 3 0 gly glycoproteins 284:296 arg1 glycoproteins 284:296 vesicle surface glycoproteins 268:296 vesicle surface glycoproteins 268:296 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 12 1 theme urinary 1446:1452 arg1 exosomes 1454:1461 urinary exosomes 1446:1461 urinary exosomes 1446:1461 We have identified the N-glycoproteome of urinary exosomes. 25452312 5 2 theme parent 424:429 arg1 cells 431:435 the parent cells 420:435 the parent cells 420:435 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 8 3 theme other 951:955 arg1 lines 982:986 T cell lines 975:986 T cell lines 975:986 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 8 3 theme other 951:955 arg1 milk 998:1001 human milk 992:1001 human milk 992:1001 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 8 3 theme other 951:955 arg1 sources 957:963 other sources 951:963 other sources including T cell lines and human milk 951:1001 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 7 4 theme urinary 717:723 arg1 exosomes 725:732 urinary exosomes 717:732 urinary exosomes in health and disease 717:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 3 5 theme targeting 228:236 arg1 mechanism 238:246 One potential targeting mechanism 214:246 One potential targeting mechanism 214:246 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 0 6 link N-linked 0:7 arg1 glycoproteomics 14:28 N-linked (N-) glycoproteomics 0:28 N-linked (N-) glycoproteomics of urinary exosomes 0:48 N-linked (N-) glycoproteomics of urinary exosomes. 25452312 10 7 theme glycosylation 1189:1201 arg1 site 1203:1206 glycosylation site 1189:1206 glycosylation site 1189:1206 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 17 8 theme urinary 2031:2037 arg1 exosomes 2039:2046 urinary exosomes 2031:2046 urinary exosomes 2031:2046 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 8 9 theme cell 977:980 arg1 lines 982:986 T cell lines 975:986 T cell lines 975:986 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 6 10 theme urinary 580:586 arg1 exosomes 588:595 urinary exosomes 580:595 urinary exosomes 580:595 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 4 11 gly glycopeptide 331:342 arg2 glycopeptide 331:342 glycopeptide 331:342 glycopeptide 331:342 This makes the glycopeptide analysis of exosomes important. 25452312 7 12 theme proteomic 687:695 arg1 characterization 697:712 proteomic characterization 687:712 proteomic characterization of urinary exosomes in health and disease 687:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 10 13 gly N-glycopeptides 1098:1112 arg2 N-glycopeptides 1098:1112 These enriched N-glycopeptides 1083:1112 These enriched N-glycopeptides 1083:1112 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 5 14 theme good 460:463 arg1 source 465:470 a good source 458:470 a good source of biomarkers for urological and other diseases 458:518 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 5 14 theme good 460:463 arg1 they 449:452 they 449:452 they 449:452 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 15 15 theme released 1806:1813 arg1 N-glycans 1815:1823 released N-glycans 1806:1823 released N-glycans 1806:1823 A corresponding glycomic analysis with released N-glycans was also performed. 25452312 17 16 theme first 1983:1987 arg1 analysis 2000:2007 the first systematic analysis 1979:2007 the first systematic analysis of N-glycoproteome of urinary exosomes 1979:2046 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 17 16 theme first 1983:1987 arg1 This 1971:1974 This 1971:1974 This 1971:1974 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 7 17 from exosomes 725:732 arg1 disease 748:754 disease 748:754 disease 748:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 7 17 from exosomes 725:732 arg1 health 737:742 health 737:742 health 737:742 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 10 18 theme dissociation 1232:1243 arg1 data 1267:1270 collision-induced dissociation MS/MS (CID-tandem MS) data 1214:1270 collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId 1214:1330 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 10 18 theme dissociation 1232:1243 arg1 MS 1263:1264 CID-tandem MS 1252:1264 CID-tandem MS 1252:1264 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 10 19 gly glycosylation 1189:1201 arg2 site 1203:1206 glycosylation site 1189:1206 glycosylation site 1189:1206 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 7 20 from health 737:742 arg1 characterization 697:712 proteomic characterization 687:712 proteomic characterization of urinary exosomes in health and disease 687:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 14 21 theme N-glycopeptides 1611:1625 arg1 sequences 1592:1600 The peptide sequences 1580:1600 The peptide sequences of these N-glycopeptides 1580:1625 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 7 22 from disease 748:754 arg1 characterization 697:712 proteomic characterization 687:712 proteomic characterization of urinary exosomes in health and disease 687:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 10 23 theme collision-induced 1214:1230 arg1 data 1267:1270 collision-induced dissociation MS/MS (CID-tandem MS) data 1214:1270 collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId 1214:1330 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 10 23 theme collision-induced 1214:1230 arg1 MS 1263:1264 CID-tandem MS 1252:1264 CID-tandem MS 1252:1264 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 8 24 theme profile 833:839 arg1 analysis 800:807 no systematic analysis 786:807 no systematic analysis of their glycoproteomic profile 786:839 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 14 25 theme peptide 1584:1590 arg1 sequences 1592:1600 The peptide sequences 1580:1600 The peptide sequences of these N-glycopeptides 1580:1625 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 6 26 theme organ 639:643 arg1 systems 645:651 renal and systemic organ systems 620:651 renal and systemic organ systems 620:651 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 16 27 theme nonmodified 1869:1879 arg1 compositions 1890:1901 66 unique nonmodified N-glycan compositions 1859:1901 66 unique nonmodified N-glycan compositions 1859:1901 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 7 28 from studies 676:682 arg1 characterization 697:712 proteomic characterization 687:712 proteomic characterization of urinary exosomes in health and disease 687:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 10 29 theme MS/MS 1245:1249 arg1 data 1267:1270 collision-induced dissociation MS/MS (CID-tandem MS) data 1214:1270 collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId 1214:1330 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 10 29 theme MS/MS 1245:1249 arg1 MS 1263:1264 CID-tandem MS 1252:1264 CID-tandem MS 1252:1264 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 13 30 gly glycoproteins 1539:1551 arg1 glycoproteins 1539:1551 37 glycoproteins 1536:1551 37 glycoproteins 1536:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 0 31 theme N-linked 0:7 arg1 glycoproteomics 14:28 N-linked (N-) glycoproteomics 0:28 N-linked (N-) glycoproteomics of urinary exosomes 0:48 N-linked (N-) glycoproteomics of urinary exosomes. 25452312 8 32 theme conserved 881:889 arg1 signature 898:906 a conserved glycan signature 879:906 a conserved glycan signature 879:906 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 10 33 theme available 1298:1306 arg1 software 1308:1315 a publicly available software 1287:1315 a publicly available software GlycopeptideId 1287:1330 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 0 34 theme N- 10:11 arg1 glycoproteomics 14:28 N-linked (N-) glycoproteomics 0:28 N-linked (N-) glycoproteomics of urinary exosomes 0:48 N-linked (N-) glycoproteomics of urinary exosomes. 25452312 13 35 gly N-glycosylation 1501:1515 arg2 sites 1517:1521 51 N-glycosylation sites 1498:1521 51 N-glycosylation sites belonging to 37 glycoproteins 1498:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 13 35 gly N-glycosylation 1501:1515 arg2 51 1498:1499 51 1498:1499 51 1498:1499 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 8 36 from urine 941:945 arg1 exosomes 927:934 exosomes 927:934 exosomes from urine and other sources including T cell lines and human milk 927:1001 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 9 37 gly N-glycopeptides 1046:1060 arg1 vesicles 1073:1080 these vesicles 1067:1080 these vesicles 1067:1080 Here, we have enriched and identified the N-glycopeptides from these vesicles. 25452312 9 37 gly N-glycopeptides 1046:1060 arg2 N-glycopeptides 1046:1060 the N-glycopeptides 1042:1060 the N-glycopeptides from these vesicles 1042:1080 Here, we have enriched and identified the N-glycopeptides from these vesicles. 25452312 4 38 theme glycopeptide 331:342 arg1 analysis 344:351 the glycopeptide analysis 327:351 the glycopeptide analysis of exosomes 327:363 This makes the glycopeptide analysis of exosomes important. 25452312 8 39 dep carried 850:856 arg1 whereas 871:877 whereas 871:877 whereas 871:877 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 0 40 theme urinary 33:39 arg1 exosomes 41:48 urinary exosomes 33:48 urinary exosomes 33:48 N-linked (N-) glycoproteomics of urinary exosomes. 25452312 2 41 theme specific 167:174 arg1 cells 176:180 specific cells 167:180 specific cells modulating their functionality 167:211 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 25452312 6 42 theme urine 535:539 arg1 easy 555:558 easy 555:558 easy 555:558 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 6 42 theme urine 535:539 arg1 collection 541:550 the urine collection 531:550 the urine collection 531:550 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 10 43 theme glycan 1154:1159 arg1 composition 1161:1171 glycan composition 1154:1171 glycan composition 1154:1171 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 12 44 theme exosomes 1454:1461 arg1 N-glycoproteome 1427:1441 the N-glycoproteome 1423:1441 the N-glycoproteome of urinary exosomes 1423:1461 We have identified the N-glycoproteome of urinary exosomes. 25452312 7 45 theme exosomes 725:732 arg1 characterization 697:712 proteomic characterization 687:712 proteomic characterization of urinary exosomes in health and disease 687:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 17 46 theme exosomes 2039:2046 arg1 N-glycoproteome 2012:2026 N-glycoproteome 2012:2026 N-glycoproteome of urinary exosomes 2012:2046 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 2 47 theme urinary 92:98 arg1 tract 100:104 the urinary tract 88:104 the urinary tract 88:104 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 25452312 5 48 theme cells 431:435 arg1 state 411:415 the physiological state 393:415 the physiological state of the parent cells 393:435 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 8 49 theme T 975:975 arg1 lines 982:986 T cell lines 975:986 T cell lines 975:986 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 5 50 theme biomarkers 475:484 arg1 source 465:470 a good source 458:470 a good source of biomarkers for urological and other diseases 458:518 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 5 50 theme biomarkers 475:484 arg1 they 449:452 they 449:452 they 449:452 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 15 51 theme glycomic 1783:1790 arg1 analysis 1792:1799 A corresponding glycomic analysis 1767:1799 A corresponding glycomic analysis with released N-glycans 1767:1823 A corresponding glycomic analysis with released N-glycans was also performed. 25452312 11 52 theme Released 1333:1340 arg1 glycans 1342:1348 Released glycans 1333:1348 Released glycans from the same sample 1333:1369 Released glycans from the same sample was also analyzed with MALDI-MS. 25452312 16 53 theme sulfated/phosphorylated 1922:1944 arg1 glycans 1946:1952 13 sulfated/phosphorylated glycans 1919:1952 13 sulfated/phosphorylated glycans 1919:1952 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 13 54 located found 1558:1562 arg1 total 1467:1471 total 1467:1471 total 1467:1471 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 13 54 located found 1558:1562 arg2 N-glycopeptides 1477:1491 126 N-glycopeptides 1473:1491 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins 1473:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 13 54 located found 1558:1562 arg1 results 1571:1577 our results 1567:1577 our results 1567:1577 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 3 55 theme potential 218:226 arg1 mechanism 238:246 One potential targeting mechanism 214:246 One potential targeting mechanism 214:246 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 8 56 theme human 992:996 arg1 milk 998:1001 human milk 992:1001 human milk 992:1001 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 3 57 theme vesicle 268:274 arg1 glycoproteins 284:296 vesicle surface glycoproteins 268:296 vesicle surface glycoproteins 268:296 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 11 58 theme same 1359:1362 arg1 sample 1364:1369 the same sample 1355:1369 the same sample 1355:1369 Released glycans from the same sample was also analyzed with MALDI-MS. 25452312 17 59 theme systematic 1989:1998 arg1 analysis 2000:2007 the first systematic analysis 1979:2007 the first systematic analysis of N-glycoproteome of urinary exosomes 1979:2046 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 17 59 theme systematic 1989:1998 arg1 This 1971:1974 This 1971:1974 This 1971:1974 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 14 60 theme glycan 1667:1672 arg1 composition 1674:1684 their glycan composition 1661:1684 their glycan composition (for 125 N-glycopeptides) 1661:1710 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 3 61 theme target 302:307 arg1 cells 309:313 target cells 302:313 target cells 302:313 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 15 62 theme corresponding 1769:1781 arg1 analysis 1792:1799 A corresponding glycomic analysis 1767:1799 A corresponding glycomic analysis with released N-glycans 1767:1823 A corresponding glycomic analysis with released N-glycans was also performed. 25452312 2 63 theme Epithelial 64:73 arg1 cells 75:79 Epithelial cells 64:79 Epithelial cells lining the urinary tract 64:104 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 25452312 7 64 theme multiple 667:674 arg1 studies 676:682 multiple studies 667:682 multiple studies on proteomic characterization of urinary exosomes in health and disease 667:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 16 65 theme unique 1862:1867 arg1 compositions 1890:1901 66 unique nonmodified N-glycan compositions 1859:1901 66 unique nonmodified N-glycan compositions 1859:1901 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 13 66 from sites 1517:1521 arg1 N-glycopeptides 1477:1491 126 N-glycopeptides 1473:1491 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins 1473:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 6 67 theme systemic 630:637 arg1 systems 645:651 renal and systemic organ systems 620:651 renal and systemic organ systems 620:651 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 3 68 theme surface 276:282 arg1 glycoproteins 284:296 vesicle surface glycoproteins 268:296 vesicle surface glycoproteins 268:296 One potential targeting mechanism is adhesion between vesicle surface glycoproteins and target cells. 25452312 16 69 located found 1964:1968 arg1 addition 1910:1917 addition 1910:1917 addition 1910:1917 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 16 69 located found 1964:1968 arg2 glycans 1946:1952 13 sulfated/phosphorylated glycans 1919:1952 13 sulfated/phosphorylated glycans 1919:1952 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 5 70 theme other 505:509 arg1 diseases 511:518 other diseases 505:518 other diseases 505:518 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 16 71 theme N-glycan 1881:1888 arg1 compositions 1890:1901 66 unique nonmodified N-glycan compositions 1859:1901 66 unique nonmodified N-glycan compositions 1859:1901 We identified 66 unique nonmodified N-glycan compositions and in addition 13 sulfated/phosphorylated glycans were also found. 25452312 8 72 theme glycoproteomic 818:831 arg1 profile 833:839 their glycoproteomic profile 812:839 their glycoproteomic profile 812:839 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 5 73 theme physiological 397:409 arg1 state 411:415 the physiological state 393:415 the physiological state of the parent cells 393:435 Exosomes reflect the physiological state of the parent cells; therefore, they are a good source of biomarkers for urological and other diseases. 25452312 2 74 dep cells 75:79 arg1 lining 81:86 lining 81:86 lining 81:86 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 25452312 4 75 theme exosomes 356:363 arg1 analysis 344:351 the glycopeptide analysis 327:351 the glycopeptide analysis of exosomes 327:363 This makes the glycopeptide analysis of exosomes important. 25452312 6 76 theme renal 620:624 arg1 systems 645:651 renal and systemic organ systems 620:651 renal and systemic organ systems 620:651 Moreover, the urine collection is easy and noninvasive and urinary exosomes give information about renal and systemic organ systems. 25452312 15 77 with analysis 1792:1799 arg1 N-glycans 1815:1823 released N-glycans 1806:1823 released N-glycans 1806:1823 A corresponding glycomic analysis with released N-glycans was also performed. 25452312 11 78 from sample 1364:1369 arg1 glycans 1342:1348 Released glycans 1333:1348 Released glycans from the same sample 1333:1369 Released glycans from the same sample was also analyzed with MALDI-MS. 25452312 14 79 gly N-glycopeptides 1611:1625 arg2 N-glycopeptides 1611:1625 these N-glycopeptides 1605:1625 these N-glycopeptides 1605:1625 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 8 80 theme systematic 789:798 arg1 analysis 800:807 no systematic analysis 786:807 no systematic analysis of their glycoproteomic profile 786:839 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 14 81 gly N-glycopeptides 1695:1709 arg2 N-glycopeptides 1695:1709 125 N-glycopeptides 1691:1709 125 N-glycopeptides 1691:1709 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 7 82 from characterization 697:712 arg1 disease 748:754 disease 748:754 disease 748:754 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 7 82 from characterization 697:712 arg1 health 737:742 health 737:742 health 737:742 Accordingly, multiple studies on proteomic characterization of urinary exosomes in health and disease have been published. 25452312 13 83 theme N-glycosylation 1501:1515 arg1 sites 1517:1521 51 N-glycosylation sites 1498:1521 51 N-glycosylation sites belonging to 37 glycoproteins 1498:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 10 84 theme software 1308:1315 arg1 GlycopeptideId 1317:1330 a publicly available software GlycopeptideId 1287:1330 a publicly available software GlycopeptideId 1287:1330 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 10 85 theme enriched 1089:1096 arg1 N-glycopeptides 1098:1112 These enriched N-glycopeptides 1083:1112 These enriched N-glycopeptides 1083:1112 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 8 86 theme glycan 891:896 arg1 signature 898:906 a conserved glycan signature 879:906 a conserved glycan signature 879:906 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 0 87 theme exosomes 41:48 arg1 glycoproteomics 14:28 N-linked (N-) glycoproteomics 0:28 N-linked (N-) glycoproteomics of urinary exosomes 0:48 N-linked (N-) glycoproteomics of urinary exosomes. 25452312 10 88 theme peptide 1136:1142 arg1 sequence 1144:1151 their peptide sequence 1130:1151 their peptide sequence 1130:1151 These enriched N-glycopeptides were solved for their peptide sequence, glycan composition, structure, and glycosylation site using collision-induced dissociation MS/MS (CID-tandem MS) data interpreted by a publicly available software GlycopeptideId. 25452312 9 89 from vesicles 1073:1080 arg1 N-glycopeptides 1046:1060 the N-glycopeptides 1042:1060 the N-glycopeptides from these vesicles 1042:1080 Here, we have enriched and identified the N-glycopeptides from these vesicles. 25452312 8 90 from sources 957:963 arg1 exosomes 927:934 exosomes 927:934 exosomes from urine and other sources including T cell lines and human milk 927:1001 However, no systematic analysis of their glycoproteomic profile has been carried out to date, whereas a conserved glycan signature has been found for exosomes from urine and other sources including T cell lines and human milk. 25452312 14 91 gly N-glycopeptides 1735:1749 arg2 N-glycopeptides 1735:1749 87 N-glycopeptides 1732:1749 87 N-glycopeptides 1732:1749 The peptide sequences of these N-glycopeptides were identified unambiguously and their glycan composition (for 125 N-glycopeptides) and structures (for 87 N-glycopeptides) were proposed. 25452312 17 92 theme N-glycoproteome 2012:2026 arg1 analysis 2000:2007 the first systematic analysis 1979:2007 the first systematic analysis of N-glycoproteome of urinary exosomes 1979:2046 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 17 92 theme N-glycoproteome 2012:2026 arg1 This 1971:1974 This 1971:1974 This 1971:1974 This is the first systematic analysis of N-glycoproteome of urinary exosomes. 25452312 13 93 gly N-glycopeptides 1477:1491 arg1 sites 1517:1521 51 N-glycosylation sites 1498:1521 51 N-glycosylation sites belonging to 37 glycoproteins 1498:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 13 93 gly N-glycopeptides 1477:1491 arg2 N-glycopeptides 1477:1491 126 N-glycopeptides 1473:1491 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins 1473:1551 In total 126 N-glycopeptides from 51 N-glycosylation sites belonging to 37 glycoproteins were found in our results. 25452312 2 94 theme urinary 114:120 arg1 exosomes 122:129 urinary exosomes 114:129 urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality 114:211 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 25452312 2 94 theme urinary 114:120 arg1 nm 139:140 40-100 nm 132:140 40-100 nm 132:140 Epithelial cells lining the urinary tract secrete urinary exosomes (40-100 nm) that can be targeted to specific cells modulating their functionality. 24824609 8 0 theme Reactivity 1146:1155 arg1 assays 1157:1162 Reactivity assays 1146:1162 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin 1146:1249 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 12 1 theme new 1807:1809 arg1 C-sheet 1778:1784 the C-sheet 1774:1784 the C-sheet of antithrombin 1774:1800 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 12 1 theme new 1807:1809 arg1 region 1811:1816 a new region 1805:1816 a new region that is relevant for proper maturation of the N-glycans 1805:1872 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 12 1 theme new 1807:1809 arg1 relevant 1826:1833 relevant 1826:1833 relevant 1826:1833 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 10 2 theme target 1530:1535 arg1 proteases 1537:1545 the target proteases 1526:1545 the target proteases 1526:1545 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 3 3 theme antithrombin 664:675 arg1 glycosylation 647:659 the glycosylation 643:659 the glycosylation of antithrombin 643:675 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 13 4 theme heparin 1975:1981 arg1 affinity 1983:1990 the reduced heparin affinity 1963:1990 the reduced heparin affinity 1963:1990 Therefore, pleiotropic mutations lead to glycosylation defects that are responsible for the reduced heparin affinity. 24824609 11 5 theme impaired 1662:1669 arg1 activity 1682:1689 impaired inhibitory activity 1662:1689 impaired inhibitory activity that was partially compensated for by activation with heparin 1662:1751 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 7 6 theme Heparin 1086:1092 arg1 affinity 1094:1101 Heparin affinity 1086:1101 Heparin affinity 1086:1101 Heparin affinity was evaluated from intrinsic fluorescence. 24824609 4 7 theme C-sheet 841:847 arg1 strands 826:832 the strands 822:832 the strands of the C-sheet 822:847 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 11 8 theme wild-type 1634:1642 arg1 antithrombin 1644:1655 wild-type antithrombin 1634:1655 wild-type antithrombin 1634:1655 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 4 9 with patients 757:764 arg1 deficiency 784:793 antithrombin deficiency 771:793 antithrombin deficiency 771:793 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 0 10 theme mutations 104:112 arg1 relevance 79:87 functional relevance 68:87 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 0:113 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 1 11 theme antithrombin 195:206 arg1 deficiency 208:217 antithrombin deficiency 195:217 antithrombin deficiency 195:217 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 0 12 from antithrombin 54:65 arg1 Role 0:3 Role 0:3 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 0:113 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 4 13 from one 815:817 arg1 located 799:805 located 799:805 located 799:805 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 10 14 with affinity 1479:1486 arg1 proteases 1537:1545 the target proteases 1526:1545 the target proteases 1526:1545 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 2 15 theme thrombosis 537:546 arg1 risk 529:532 the mild risk 520:532 the mild risk of thrombosis in carriers 520:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 2 15 theme thrombosis 537:546 arg1 affinity 507:514 the impaired heparin affinity 486:514 the impaired heparin affinity 486:514 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 0 16 theme N-glycans 41:49 arg1 maturation 27:36 the maturation 23:36 the maturation of N-glycans on antithrombin 23:65 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 10 17 with reactivity 1510:1519 arg1 proteases 1537:1545 the target proteases 1526:1545 the target proteases 1526:1545 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 10 18 theme abnormal 1435:1442 arg1 glycosylation 1444:1456 abnormal glycosylation 1435:1456 abnormal glycosylation 1435:1456 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 1 19 theme natural 151:157 arg1 mutants 159:165 natural mutants 151:165 natural mutants identified in patients with antithrombin deficiency 151:217 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 6 20 theme Glycoform 953:961 arg1 heterogeneity 963:975 Glycoform heterogeneity 953:975 Glycoform heterogeneity 953:975 Glycoform heterogeneity and conformational sensitivity were studied with electrophoresis, proteomic analysis, and glycomic analysis. 24824609 0 21 from Role 0:3 arg1 antithrombin 54:65 antithrombin 54:65 antithrombin 54:65 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 0 21 from Role 0:3 arg1 maturation 27:36 the maturation 23:36 the maturation of N-glycans on antithrombin 23:65 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 0 22 theme functional 68:77 arg1 relevance 79:87 functional relevance 68:87 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 0:113 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 2 23 theme impaired 490:497 arg1 affinity 507:514 the impaired heparin affinity 486:514 the impaired heparin affinity 486:514 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 0 24 from maturation 27:36 arg1 antithrombin 54:65 antithrombin 54:65 antithrombin 54:65 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 10 25 theme reduced 1463:1469 arg1 affinity 1479:1486 reduced heparin affinity 1463:1486 reduced heparin affinity 1463:1486 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 4 26 theme antithrombin 771:782 arg1 deficiency 784:793 antithrombin deficiency 771:793 antithrombin deficiency 771:793 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 4 27 theme pleiotropic 721:731 arg1 mutations 733:741 Five pleiotropic mutations 716:741 Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet 716:847 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 6 28 theme proteomic 1043:1051 arg1 analysis 1053:1060 proteomic analysis 1043:1060 proteomic analysis 1043:1060 Glycoform heterogeneity and conformational sensitivity were studied with electrophoresis, proteomic analysis, and glycomic analysis. 24824609 2 29 theme abnormal 416:423 arg1 glycosylation 425:437 an abnormal glycosylation 413:437 an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers 413:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 8 30 with assays 1157:1162 arg1 thrombin 1180:1187 thrombin 1180:1187 thrombin 1180:1187 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 8 30 with assays 1157:1162 arg1 elastase 1204:1211 neutrophil elastase 1193:1211 neutrophil elastase 1193:1211 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 8 30 with assays 1157:1162 arg1 factor Xa 1169:1177 factor Xa 1169:1177 factor Xa 1169:1177 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 1 31 theme anticoagulant 284:296 arg1 anticoagulant 284:296 this key anticoagulant 275:296 this key anticoagulant 275:296 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 31 theme anticoagulant 284:296 arg1 mechanisms 306:315 the mechanisms 302:315 the mechanisms involved in the deficiency 302:342 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 31 theme anticoagulant 284:296 arg1 regions 264:270 regions 264:270 regions 264:270 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 31 theme anticoagulant 284:296 arg1 domains 253:259 functional domains 242:259 functional domains 242:259 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 12 32 theme N-glycans 1864:1872 arg1 maturation 1846:1855 proper maturation 1839:1855 proper maturation of the N-glycans 1839:1872 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 5 33 theme Recombinant 904:914 arg1 mutants 916:922 Recombinant mutants 904:922 Recombinant mutants 904:922 Recombinant mutants were generated and purified. 24824609 11 34 contain had 1658:1660 arg2 activity 1682:1689 impaired inhibitory activity 1662:1689 impaired inhibitory activity that was partially compensated for by activation with heparin 1662:1751 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 11 34 contain had 1658:1660 arg1 variant 2 1561:1569 variant 2 1561:1569 variant 2 1561:1569 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 3 35 theme mutations 630:638 arg1 effects 589:595 the effects 585:595 the effects of different natural pleiotropic mutations on the glycosylation of antithrombin 585:675 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 3 35 theme mutations 630:638 arg1 effects 698:704 their functional effects 681:704 their functional effects 681:704 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 2 36 theme pleiotropic 444:454 arg1 K241E 464:468 K241E 464:468 K241E 464:468 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 2 36 theme pleiotropic 444:454 arg1 mutant 456:461 a pleiotropic mutant 442:461 a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers 442:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 3 37 theme natural 610:616 arg1 mutations 630:638 different natural pleiotropic mutations 600:638 different natural pleiotropic mutations 600:638 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 1 38 with patients 181:188 arg1 deficiency 208:217 antithrombin deficiency 195:217 antithrombin deficiency 195:217 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 11 39 theme similar 1577:1583 arg1 mobility 1601:1608 similar electrophoretic mobility 1577:1608 similar electrophoretic mobility 1577:1608 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 10 40 gly glycosylation 1444:1456 arg1 Variant 1 1419:1427 Variant 1 1419:1427 Variant 1 1419:1427 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 3 41 gly glycosylation 647:659 arg1 antithrombin 664:675 antithrombin 664:675 antithrombin 664:675 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 4 42 dep selected 854:861 arg1 F402L 885:889 F402L 885:889 F402L 885:889 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 4 42 dep selected 854:861 arg1 P429L 896:900 P429L 896:900 P429L 896:900 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 4 42 dep selected 854:861 arg1 M315K 878:882 M315K 878:882 M315K 878:882 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 4 42 dep selected 854:861 arg1 M251I 871:875 M251I 871:875 M251I 871:875 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 0 43 dep Role 0:3 arg1 relevance 79:87 functional relevance 68:87 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 0:113 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 3 44 theme functional 687:696 arg1 effects 698:704 their functional effects 681:704 their functional effects 681:704 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 11 45 theme heparin 1614:1620 arg1 affinity 1622:1629 heparin affinity 1614:1629 heparin affinity 1614:1629 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 6 46 theme glycomic 1067:1074 arg1 analysis 1076:1083 glycomic analysis 1067:1083 glycomic analysis 1067:1083 Glycoform heterogeneity and conformational sensitivity were studied with electrophoresis, proteomic analysis, and glycomic analysis. 24824609 9 47 dep RESULTS 1272:1278 arg1 showed 1396:1401 showed 1396:1401 showed two glycoforms 1396:1416 RESULTS AND CONCLUSIONS: Pleiotropic mutants, except for that with the M315K mutation, which affects a non-exposed residue, showed two glycoforms. 24824609 12 48 theme antithrombin 1789:1800 arg1 C-sheet 1778:1784 the C-sheet 1774:1784 the C-sheet of antithrombin 1774:1800 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 12 48 theme antithrombin 1789:1800 arg1 region 1811:1816 a new region 1805:1816 a new region that is relevant for proper maturation of the N-glycans 1805:1872 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 12 48 theme antithrombin 1789:1800 arg1 relevant 1826:1833 relevant 1826:1833 relevant 1826:1833 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 11 49 with activation 1729:1738 arg1 heparin 1745:1751 heparin 1745:1751 heparin 1745:1751 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 1 50 theme mutants 159:165 arg1 characterization 131:146 The characterization 127:146 The characterization of natural mutants identified in patients with antithrombin deficiency 127:217 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 4 51 theme strands 826:832 arg1 one 815:817 one 815:817 one 815:817 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 4 51 theme strands 826:832 arg1 strands 826:832 the strands 822:832 the strands of the C-sheet 822:847 METHODS: Five pleiotropic mutations identified in patients with antithrombin deficiency and located at each one of the strands of the C-sheet were selected (K241E, M251I, M315K, F402L, and P429L). 24824609 3 52 from effects 698:704 arg1 glycosylation 647:659 the glycosylation 643:659 the glycosylation of antithrombin 643:675 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 11 53 theme inhibitory 1671:1680 arg1 activity 1682:1689 impaired inhibitory activity 1662:1689 impaired inhibitory activity that was partially compensated for by activation with heparin 1662:1751 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 12 54 theme proper 1839:1844 arg1 maturation 1846:1855 proper maturation 1839:1855 proper maturation of the N-glycans 1839:1872 Our results suggest the C-sheet of antithrombin as a new region that is relevant for proper maturation of the N-glycans. 24824609 8 55 theme heparin 1243:1249 arg1 absence 1232:1238 absence 1232:1238 absence 1232:1238 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 8 55 theme heparin 1243:1249 arg1 presence 1220:1227 presence 1220:1227 presence 1220:1227 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 13 56 theme reduced 1967:1973 arg1 affinity 1983:1990 the reduced heparin affinity 1963:1990 the reduced heparin affinity 1963:1990 Therefore, pleiotropic mutations lead to glycosylation defects that are responsible for the reduced heparin affinity. 24824609 8 57 theme neutrophil 1193:1202 arg1 elastase 1204:1211 neutrophil elastase 1193:1211 neutrophil elastase 1193:1211 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 2 58 theme mild 524:527 arg1 risk 529:532 the mild risk 520:532 the mild risk of thrombosis in carriers 520:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 1 59 theme clinical 370:377 arg1 prognosis 379:387 the clinical prognosis 366:387 the clinical prognosis 366:387 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 10 60 contain had 1459:1461 arg1 Variant 1 1419:1427 Variant 1 1419:1427 Variant 1 1419:1427 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 10 60 contain had 1459:1461 arg2 reactivity 1510:1519 severely affected reactivity 1492:1519 severely affected reactivity with the target proteases 1492:1545 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 10 60 contain had 1459:1461 arg2 affinity 1479:1486 reduced heparin affinity 1463:1486 reduced heparin affinity 1463:1486 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 2 61 from glycosylation 425:437 arg1 K241E 464:468 K241E 464:468 K241E 464:468 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 2 61 from glycosylation 425:437 arg1 mutant 456:461 a pleiotropic mutant 442:461 a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers 442:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 13 62 theme glycosylation 1916:1928 arg1 defects 1930:1936 glycosylation defects 1916:1936 glycosylation defects that are responsible for the reduced heparin affinity 1916:1990 Therefore, pleiotropic mutations lead to glycosylation defects that are responsible for the reduced heparin affinity. 24824609 2 63 from affinity 507:514 arg1 carriers 551:558 carriers 551:558 carriers 551:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 6 64 theme conformational 981:994 arg1 sensitivity 996:1006 conformational sensitivity 981:1006 conformational sensitivity 981:1006 Glycoform heterogeneity and conformational sensitivity were studied with electrophoresis, proteomic analysis, and glycomic analysis. 24824609 13 65 theme pleiotropic 1886:1896 arg1 mutations 1898:1906 pleiotropic mutations 1886:1906 pleiotropic mutations 1886:1906 Therefore, pleiotropic mutations lead to glycosylation defects that are responsible for the reduced heparin affinity. 24824609 9 66 theme Pleiotropic 1297:1307 arg1 mutants 1309:1315 Pleiotropic mutants 1297:1315 Pleiotropic mutants 1297:1315 RESULTS AND CONCLUSIONS: Pleiotropic mutants, except for that with the M315K mutation, which affects a non-exposed residue, showed two glycoforms. 24824609 2 67 theme heparin 499:505 arg1 affinity 507:514 the impaired heparin affinity 486:514 the impaired heparin affinity 486:514 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 0 68 theme pleiotropic 92:102 arg1 mutations 104:112 pleiotropic mutations 92:112 pleiotropic mutations 92:112 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 10 69 with Variant 1 1419:1427 arg1 glycosylation 1444:1456 abnormal glycosylation 1435:1456 abnormal glycosylation 1435:1456 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 3 70 from effects 589:595 arg1 glycosylation 647:659 the glycosylation 643:659 the glycosylation of antithrombin 643:675 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 0 71 theme C-sheet 12:18 arg1 Role 0:3 Role 0:3 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 0:113 Role of the C-sheet in the maturation of N-glycans on antithrombin: functional relevance of pleiotropic mutations. 24824609 1 72 theme key 280:282 arg1 anticoagulant 284:296 this key anticoagulant 275:296 this key anticoagulant 275:296 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 11 73 with variant 2 1561:1569 arg1 mobility 1601:1608 similar electrophoretic mobility 1577:1608 similar electrophoretic mobility 1577:1608 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 11 73 with variant 2 1561:1569 arg1 affinity 1622:1629 heparin affinity 1614:1629 heparin affinity 1614:1629 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 8 74 dep presence 1220:1227 arg1 the 1216:1218 the 1216:1218 the 1216:1218 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 3 75 theme pleiotropic 618:628 arg1 mutations 630:638 different natural pleiotropic mutations 600:638 different natural pleiotropic mutations 600:638 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 9 76 theme M315K 1343:1347 arg1 mutation 1349:1356 the M315K mutation 1339:1356 the M315K mutation 1339:1356 RESULTS AND CONCLUSIONS: Pleiotropic mutants, except for that with the M315K mutation, which affects a non-exposed residue, showed two glycoforms. 24824609 10 77 theme heparin 1471:1477 arg1 affinity 1479:1486 reduced heparin affinity 1463:1486 reduced heparin affinity 1463:1486 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 3 78 theme different 600:608 arg1 mutations 630:638 different natural pleiotropic mutations 600:638 different natural pleiotropic mutations 600:638 OBJECTIVES: To evaluate the effects of different natural pleiotropic mutations on the glycosylation of antithrombin and their functional effects. 24824609 11 79 theme electrophoretic 1585:1599 arg1 mobility 1601:1608 similar electrophoretic mobility 1577:1608 similar electrophoretic mobility 1577:1608 In contrast, variant 2, with similar electrophoretic mobility and heparin affinity to wild-type antithrombin, had impaired inhibitory activity that was partially compensated for by activation with heparin. 24824609 7 80 theme intrinsic 1122:1130 arg1 fluorescence 1132:1143 intrinsic fluorescence 1122:1143 intrinsic fluorescence 1122:1143 Heparin affinity was evaluated from intrinsic fluorescence. 24824609 1 81 theme mechanisms 306:315 arg1 anticoagulant 284:296 this key anticoagulant 275:296 this key anticoagulant 275:296 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 81 theme mechanisms 306:315 arg1 mechanisms 306:315 the mechanisms 302:315 the mechanisms involved in the deficiency 302:342 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 81 theme mechanisms 306:315 arg1 regions 264:270 regions 264:270 regions 264:270 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 81 theme mechanisms 306:315 arg1 domains 253:259 functional domains 242:259 functional domains 242:259 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 2 82 gly glycosylation 425:437 arg1 K241E 464:468 K241E 464:468 K241E 464:468 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 2 82 gly glycosylation 425:437 arg1 mutant 456:461 a pleiotropic mutant 442:461 a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers 442:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 10 83 theme affected 1501:1508 arg1 reactivity 1510:1519 severely affected reactivity 1492:1519 severely affected reactivity with the target proteases 1492:1545 Variant 1, with abnormal glycosylation, had reduced heparin affinity and severely affected reactivity with the target proteases. 24824609 9 84 theme non-exposed 1375:1385 arg1 residue 1387:1393 a non-exposed residue 1373:1393 a non-exposed residue 1373:1393 RESULTS AND CONCLUSIONS: Pleiotropic mutants, except for that with the M315K mutation, which affects a non-exposed residue, showed two glycoforms. 24824609 8 85 from assays 1157:1162 arg1 absence 1232:1238 absence 1232:1238 absence 1232:1238 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 8 85 from assays 1157:1162 arg1 presence 1220:1227 presence 1220:1227 presence 1220:1227 Reactivity assays with factor Xa, thrombin and neutrophil elastase in the presence or absence of heparin were also performed. 24824609 2 86 from risk 529:532 arg1 carriers 551:558 carriers 551:558 carriers 551:558 Recently, we described an abnormal glycosylation in a pleiotropic mutant (K241E) that explained the impaired heparin affinity and the mild risk of thrombosis in carriers. 24824609 1 87 theme functional 242:251 arg1 anticoagulant 284:296 this key anticoagulant 275:296 this key anticoagulant 275:296 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 87 theme functional 242:251 arg1 mechanisms 306:315 the mechanisms 302:315 the mechanisms involved in the deficiency 302:342 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 24824609 1 87 theme functional 242:251 arg1 domains 253:259 functional domains 242:259 functional domains 242:259 BACKGROUND: The characterization of natural mutants identified in patients with antithrombin deficiency has helped to identify functional domains or regions of this key anticoagulant and the mechanisms involved in the deficiency, as well as to define the clinical prognosis. 26302170 6 0 theme unique 1099:1104 arg1 site 1114:1117 a unique binding site 1097:1117 a unique binding site 1097:1117 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 2 1 theme polyomavirus 366:377 arg1 family 379:384 the polyomavirus family 362:384 the polyomavirus family 362:384 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 6 2 theme established 1169:1179 arg1 sites 1201:1205 established sialic acid binding sites 1169:1205 established sialic acid binding sites of other polyomaviruses 1169:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 2 theme established 1169:1179 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 7 3 theme Structure-based 1232:1246 arg1 mutagenesis 1248:1258 Structure-based mutagenesis 1232:1258 Structure-based mutagenesis of sialic acid-binding residues 1232:1290 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 1 4 theme immunocompromised 292:308 arg1 patients 310:317 severely immunocompromised patients 283:317 severely immunocompromised patients 283:317 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 5 5 theme protein 890:896 arg1 VP1 898:900 the major capsid protein VP1 873:900 the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 873:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 9 6 from location 1860:1867 arg1 variable 1785:1792 variable 1785:1792 variable 1785:1792 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 7 7 theme interaction 1423:1433 arg1 relevance 1389:1397 the physiological relevance 1371:1397 the physiological relevance of the TSPyV VP1-glycan interaction 1371:1433 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 7 8 theme TSPyV 1406:1410 arg1 interaction 1423:1433 the TSPyV VP1-glycan interaction 1402:1433 the TSPyV VP1-glycan interaction 1402:1433 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 7 9 theme pseudovirus 1334:1344 arg1 infection 1346:1354 pseudovirus infection 1334:1354 pseudovirus infection 1334:1354 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 5 10 dep trisaccharides 992:1005 arg1 α2,6-sialyllactose 1017:1034 α2,6-sialyllactose 1017:1034 α2,6-sialyllactose 1017:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 10 dep trisaccharides 992:1005 arg1 α2,3- 1007:1011 α2,3- 1007:1011 α2,3- 1007:1011 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 11 theme major 877:881 arg1 VP1 898:900 the major capsid protein VP1 873:900 the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 873:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 8 12 theme N- 1488:1489 arg1 inhibitors 1474:1483 inhibitors 1474:1483 inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis 1474:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 6 13 theme other 1210:1214 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 4 14 theme infection 724:732 arg1 studies 734:740 pseudovirus infection studies 712:740 pseudovirus infection studies 712:740 We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. 26302170 3 15 theme structure-function 596:613 arg1 approach 615:622 a structure-function approach 594:622 a structure-function approach to shed light on the first steps of TSPyV infection 594:674 Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. 26302170 5 16 from structures 859:868 arg1 complex 914:920 complex 914:920 complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 914:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 17 theme different 933:941 arg1 glycans 943:949 three different glycans 927:949 three different glycans 927:949 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 8 18 theme O-linked 1492:1499 arg1 glycosylation 1501:1513 O-linked glycosylation 1492:1513 O-linked glycosylation 1492:1513 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 7 19 theme physiological 1375:1387 arg1 relevance 1389:1397 the physiological relevance 1371:1397 the physiological relevance of the TSPyV VP1-glycan interaction 1371:1433 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 5 20 from VP1 898:900 arg1 complex 914:920 complex 914:920 complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 914:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 6 21 theme terminal 1041:1048 arg1 acid 1057:1060 The terminal sialic acid 1037:1060 The terminal sialic acid of all three glycans 1037:1081 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 2 22 theme intracellular 504:516 arg1 events 530:535 intracellular trafficking events 504:535 intracellular trafficking events 504:535 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 6 23 theme acid 1188:1191 arg1 sites 1201:1205 established sialic acid binding sites 1169:1205 established sialic acid binding sites of other polyomaviruses 1169:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 23 theme acid 1188:1191 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 4 24 theme cell 695:698 arg1 binding 700:706 cell binding 695:706 cell binding 695:706 We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. 26302170 8 25 theme glycosphingolipid 1520:1536 arg1 synthesis 1538:1546 glycosphingolipid synthesis 1520:1546 glycosphingolipid synthesis 1520:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 5 26 with complex 914:920 arg1 trisaccharides 992:1005 the linear trisaccharides α2,3- and α2,6-sialyllactose 981:1034 the linear trisaccharides α2,3- and α2,6-sialyllactose 981:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 26 with complex 914:920 arg1 glycan 969:974 the branched GM1 glycan 952:974 the branched GM1 glycan 952:974 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 26 with complex 914:920 arg1 glycans 943:949 three different glycans 927:949 three different glycans 927:949 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 27 from complex 914:920 arg1 structures 859:868 high-resolution X-ray structures 837:868 high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 837:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 9 28 from variable 1785:1792 arg1 location 1860:1867 the location 1856:1867 the location of the binding site 1856:1887 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 9 28 from variable 1785:1792 arg1 interactions 1806:1817 interactions 1806:1817 interactions with sialic acid itself 1806:1841 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 2 29 theme TSPyV 544:548 arg1 infection 550:558 TSPyV infection 544:558 TSPyV infection 544:558 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 0 30 theme Sialylated 97:106 arg1 Glycolipids 108:118 Sialylated Glycolipids 97:118 Sialylated Glycolipids 97:118 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 9 31 theme binding 1876:1882 arg1 site 1884:1887 the binding site 1872:1887 the binding site 1872:1887 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 5 32 theme linear 985:990 arg1 trisaccharides 992:1005 the linear trisaccharides α2,3- and α2,6-sialyllactose 981:1034 the linear trisaccharides α2,3- and α2,6-sialyllactose 981:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 0 33 theme Trichodysplasia 0:14 arg1 Polyomavirus 37:48 Trichodysplasia spinulosa-Associated Polyomavirus 0:48 Trichodysplasia spinulosa-Associated Polyomavirus 0:48 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 5 34 theme branched 956:963 arg1 glycan 969:974 the branched GM1 glycan 952:974 the branched GM1 glycan 952:974 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 6 35 theme glycans 1075:1081 arg1 acid 1057:1060 The terminal sialic acid 1037:1060 The terminal sialic acid of all three glycans 1037:1081 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 2 36 theme molecular 445:453 arg1 events 467:472 molecular recognition events 445:472 molecular recognition events 445:472 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 4 37 theme sialic 768:773 arg1 acids 775:779 sialic acids 768:779 sialic acids 768:779 We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. 26302170 5 38 theme X-ray 853:857 arg1 structures 859:868 high-resolution X-ray structures 837:868 high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 837:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 1 39 theme Trichodysplasia 121:135 arg1 TSPyV 172:176 TSPyV 172:176 TSPyV 172:176 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 39 theme Trichodysplasia 121:135 arg1 Polyomavirus 158:169 Trichodysplasia spinulosa-associated Polyomavirus 121:169 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) 121:177 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 2 40 theme entry 485:489 arg1 pathway 491:497 the viral entry pathway 475:497 the viral entry pathway 475:497 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 8 41 theme TSPyV 1603:1607 arg1 infection 1609:1617 TSPyV infection 1603:1617 TSPyV infection 1603:1617 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 3 42 theme TSPyV 660:664 arg1 infection 666:674 TSPyV infection 660:674 TSPyV infection 660:674 Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. 26302170 9 43 theme site 1884:1887 arg1 location 1860:1867 the location 1856:1867 the location of the binding site 1856:1887 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 9 44 theme first 1647:1651 arg1 events 1675:1680 the first molecular recognition events 1643:1680 the first molecular recognition events of cellular infection with TSPyV 1643:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 7 45 from reduction 1301:1309 arg1 attachment 1319:1328 cell attachment 1314:1328 cell attachment 1314:1328 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 7 45 from reduction 1301:1309 arg1 infection 1346:1354 pseudovirus infection 1334:1354 pseudovirus infection 1334:1354 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 8 46 link O-linked 1492:1499 arg1 glycosylation 1501:1513 O-linked glycosylation 1492:1513 O-linked glycosylation 1492:1513 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 0 47 theme Binding 67:73 arg1 Site 75:78 a Displaced Binding Site 55:78 a Displaced Binding Site on VP1 55:85 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 9 48 theme recognition 1663:1673 arg1 events 1675:1680 the first molecular recognition events 1643:1680 the first molecular recognition events of cellular infection with TSPyV 1643:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 2 49 theme members 351:357 arg1 one 335:337 one 335:337 one 335:337 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 2 49 theme members 351:357 arg1 members 351:357 the five members 342:357 the five members of the polyomavirus family 342:384 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 1 50 theme skin 251:254 arg1 spinulosa 238:246 trichodysplasia spinulosa 222:246 trichodysplasia spinulosa 222:246 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 50 theme skin 251:254 arg1 disease 256:262 a skin disease 249:262 a skin disease that can appear in severely immunocompromised patients 249:317 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 7 51 theme residues 1283:1290 arg1 mutagenesis 1248:1258 Structure-based mutagenesis 1232:1258 Structure-based mutagenesis of sialic acid-binding residues 1232:1290 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 9 52 from interactions 1806:1817 arg1 variable 1785:1792 variable 1785:1792 variable 1785:1792 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 9 53 theme infection 1694:1702 arg1 events 1675:1680 the first molecular recognition events 1643:1680 the first molecular recognition events of cellular infection with TSPyV 1643:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 7 54 theme sialic 1263:1268 arg1 residues 1283:1290 sialic acid-binding residues 1263:1290 sialic acid-binding residues 1263:1290 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 2 55 theme family 379:384 arg1 members 351:357 the five members 342:357 the five members of the polyomavirus family 342:384 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 5 56 theme capsid 883:888 arg1 VP1 898:900 the major capsid protein VP1 873:900 the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 873:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 9 57 theme receptor 1736:1743 arg1 recognition 1745:1755 receptor recognition 1736:1755 receptor recognition by polyomaviruses 1736:1773 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 1 58 attach isolated 183:190 arg2 TSPyV 172:176 TSPyV 172:176 TSPyV 172:176 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 58 attach isolated 183:190 arg1 patient 199:205 a patient 197:205 a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients 197:317 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 58 attach isolated 183:190 arg2 Polyomavirus 158:169 Trichodysplasia spinulosa-associated Polyomavirus 121:169 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) 121:177 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 5 59 theme VP1 898:900 arg1 structures 859:868 high-resolution X-ray structures 837:868 high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 837:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 1 60 theme trichodysplasia 222:236 arg1 spinulosa 238:246 trichodysplasia spinulosa 222:246 trichodysplasia spinulosa 222:246 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 60 theme trichodysplasia 222:236 arg1 disease 256:262 a skin disease 249:262 a skin disease that can appear in severely immunocompromised patients 249:317 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 7 61 theme VP1-glycan 1412:1421 arg1 interaction 1423:1433 the TSPyV VP1-glycan interaction 1402:1433 the TSPyV VP1-glycan interaction 1402:1433 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 1 62 theme spinulosa-associated 137:156 arg1 TSPyV 172:176 TSPyV 172:176 TSPyV 172:176 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 1 62 theme spinulosa-associated 137:156 arg1 Polyomavirus 158:169 Trichodysplasia spinulosa-associated Polyomavirus 121:169 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) 121:177 Trichodysplasia spinulosa-associated Polyomavirus (TSPyV) was isolated from a patient suffering from trichodysplasia spinulosa, a skin disease that can appear in severely immunocompromised patients. 26302170 8 63 theme glycosylation 1501:1513 arg1 inhibitors 1474:1483 inhibitors 1474:1483 inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis 1474:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 4 64 theme pseudovirus 712:722 arg1 studies 734:740 pseudovirus infection studies 712:740 pseudovirus infection studies 712:740 We established by cell binding and pseudovirus infection studies that TSPyV interacts with sialic acids during attachment and/or entry. 26302170 8 65 with treatments 1449:1458 arg1 inhibitors 1474:1483 inhibitors 1474:1483 inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis 1474:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 6 66 theme sialic 1050:1055 arg1 acid 1057:1060 The terminal sialic acid 1037:1060 The terminal sialic acid of all three glycans 1037:1081 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 67 theme binding 1193:1199 arg1 sites 1201:1205 established sialic acid binding sites 1169:1205 established sialic acid binding sites of other polyomaviruses 1169:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 67 theme binding 1193:1199 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 68 theme TSPyV 1122:1126 arg1 VP1 1128:1130 TSPyV VP1 1122:1130 TSPyV VP1 1122:1130 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 5 69 theme TSPyV 905:909 arg1 VP1 898:900 the major capsid protein VP1 873:900 the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 873:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 9 70 theme sialic 1824:1829 arg1 acid 1831:1834 sialic acid 1824:1834 sialic acid itself 1824:1841 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 8 71 theme synthesis 1538:1546 arg1 inhibitors 1474:1483 inhibitors 1474:1483 inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis 1474:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 6 72 theme sialic 1181:1186 arg1 sites 1201:1205 established sialic acid binding sites 1169:1205 established sialic acid binding sites of other polyomaviruses 1169:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 72 theme sialic 1181:1186 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 73 from sites 1201:1205 arg1 Å 1162:1162 18 Å 1159:1162 18 Å from established sialic acid binding sites of other polyomaviruses 1159:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 0 74 from Site 75:78 arg1 VP1 83:85 VP1 83:85 VP1 83:85 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 2 75 theme trafficking 518:528 arg1 events 530:535 intracellular trafficking events 504:535 intracellular trafficking events 504:535 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 8 76 theme important 1581:1589 arg1 role 1591:1594 an important role 1578:1594 an important role 1578:1594 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 9 77 with interactions 1806:1817 arg1 acid 1831:1834 sialic acid 1824:1834 sialic acid itself 1824:1841 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 0 78 theme spinulosa-Associated 16:35 arg1 Polyomavirus 37:48 Trichodysplasia spinulosa-Associated Polyomavirus 0:48 Trichodysplasia spinulosa-Associated Polyomavirus 0:48 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 2 79 theme recognition 455:465 arg1 events 467:472 molecular recognition events 445:472 molecular recognition events 445:472 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 5 80 theme high-resolution 837:851 arg1 structures 859:868 high-resolution X-ray structures 837:868 high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose 837:1034 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 5 81 theme GM1 965:967 arg1 glycan 969:974 the branched GM1 glycan 952:974 the branched GM1 glycan 952:974 Subsequently, we solved high-resolution X-ray structures of the major capsid protein VP1 of TSPyV in complex with three different glycans, the branched GM1 glycan, and the linear trisaccharides α2,3- and α2,6-sialyllactose. 26302170 7 82 theme cell 1314:1317 arg1 attachment 1319:1328 cell attachment 1314:1328 cell attachment 1314:1328 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 0 83 theme Displaced 57:65 arg1 Site 75:78 a Displaced Binding Site 55:78 a Displaced Binding Site on VP1 55:85 Trichodysplasia spinulosa-Associated Polyomavirus Uses a Displaced Binding Site on VP1 to Engage Sialylated Glycolipids. 26302170 3 84 used used 589:592 arg2 we 581:582 we 581:582 we 581:582 Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. 26302170 2 85 theme viral 479:483 arg1 pathway 491:497 the viral entry pathway 475:497 the viral entry pathway 475:497 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 3 86 theme first 645:649 arg1 steps 651:655 the first steps 641:655 the first steps of TSPyV infection 641:674 Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. 26302170 2 87 theme human 416:420 arg1 disease 422:428 a human disease 414:428 a human disease 414:428 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 9 88 theme molecular 1653:1661 arg1 events 1675:1680 the first molecular recognition events 1643:1680 the first molecular recognition events of cellular infection with TSPyV 1643:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 6 89 theme polyomaviruses 1216:1229 arg1 sites 1201:1205 established sialic acid binding sites 1169:1205 established sialic acid binding sites of other polyomaviruses 1169:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 6 89 theme polyomaviruses 1216:1229 arg1 polyomaviruses 1216:1229 other polyomaviruses 1210:1229 other polyomaviruses 1210:1229 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 7 90 theme acid-binding 1270:1281 arg1 residues 1283:1290 sialic acid-binding residues 1263:1290 sialic acid-binding residues 1263:1290 Structure-based mutagenesis of sialic acid-binding residues leads to reduction in cell attachment and pseudovirus infection, demonstrating the physiological relevance of the TSPyV VP1-glycan interaction. 26302170 3 91 theme infection 666:674 arg1 steps 651:655 the first steps 641:655 the first steps of TSPyV infection 641:674 Here we have used a structure-function approach to shed light on the first steps of TSPyV infection. 26302170 6 92 theme binding 1106:1112 arg1 site 1114:1117 a unique binding site 1097:1117 a unique binding site 1097:1117 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26302170 2 93 attach linked 404:409 arg1 disease 422:428 a human disease 414:428 a human disease 414:428 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 2 93 attach linked 404:409 arg2 one 335:337 one 335:337 one 335:337 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 2 93 attach linked 404:409 arg2 members 351:357 the five members 342:357 the five members of the polyomavirus family 342:384 While TSPyV is one of the five members of the polyomavirus family that are directly linked to a human disease, details about molecular recognition events, the viral entry pathway, and intracellular trafficking events during TSPyV infection remain unknown. 26302170 9 94 theme cellular 1685:1692 arg1 infection 1694:1702 cellular infection 1685:1702 cellular infection with TSPyV 1685:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 9 95 with infection 1694:1702 arg1 TSPyV 1709:1713 TSPyV 1709:1713 TSPyV 1709:1713 Our findings elucidate the first molecular recognition events of cellular infection with TSPyV and demonstrate that receptor recognition by polyomaviruses is highly variable not only in interactions with sialic acid itself, but also in the location of the binding site. 26302170 8 96 theme cells 1463:1467 arg1 treatments 1449:1458 treatments 1449:1458 treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis 1449:1546 Furthermore, treatments of cells with inhibitors of N-, O-linked glycosylation, and glycosphingolipid synthesis suggest that glycolipids play an important role during TSPyV infection. 26302170 6 97 from site 1114:1117 arg1 VP1 1128:1130 TSPyV VP1 1122:1130 TSPyV VP1 1122:1130 The terminal sialic acid of all three glycans is engaged in a unique binding site on TSPyV VP1, which is positioned about 18 Å from established sialic acid binding sites of other polyomaviruses. 26143917 9 0 from changes 1332:1338 arg1 conformation 1347:1358 conformation 1347:1358 conformation 1347:1358 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 9 0 from changes 1332:1338 arg1 state 1376:1380 protonation state 1364:1380 protonation state 1364:1380 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 7 1 theme crystals 953:960 arg1 resolution 935:944 The resolution 931:944 The resolution of the crystals of the catalytic domain 931:984 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 7 2 with forms 1070:1074 arg1 chitobiose 1097:1106 chitobiose 1097:1106 chitobiose 1097:1106 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 1 3 theme glycosyl 228:235 arg1 family 247:252 the highly conserved glycosyl hydrolase family 18 207:255 the highly conserved glycosyl hydrolase family 18 (GH18) 207:262 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 1 3 theme glycosyl 228:235 arg1 GH18 258:261 GH18 258:261 GH18 258:261 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 0 4 theme CHIT1 68:72 arg1 domain 85:90 human chitotriosidase (CHIT1) catalytic domain 45:90 human chitotriosidase (CHIT1) catalytic domain 45:90 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 6 5 theme catalytic 797:805 arg1 mechanism 807:815 the catalytic mechanism 793:815 the catalytic mechanism of GH18 chitinases 793:834 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 1 6 theme hydrolase 237:245 arg1 family 247:252 the highly conserved glycosyl hydrolase family 18 207:255 the highly conserved glycosyl hydrolase family 18 (GH18) 207:262 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 1 6 theme hydrolase 237:245 arg1 GH18 258:261 GH18 258:261 GH18 258:261 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 0 7 theme domain 85:90 arg1 mechanism 32:40 the enzymatic mechanism 18:40 the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM 18:146 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 9 8 theme hydrolysis 1471:1480 arg1 activities 1505:1514 the hydrolysis and transglycosylation activities 1467:1514 activities 1505:1514 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 4 9 theme acidic 575:580 arg1 Asp136 611:616 Asp136 611:616 Asp136 611:616 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 4 9 theme acidic 575:580 arg1 Asp138 600:605 Asp138 600:605 Asp138 600:605 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 4 9 theme acidic 575:580 arg1 Glu140 592:597 Glu140 592:597 Glu140 592:597 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 4 9 theme acidic 575:580 arg1 residues 582:589 three acidic residues 569:589 three acidic residues 569:589 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 0 10 theme catalytic 75:83 arg1 domain 85:90 human chitotriosidase (CHIT1) catalytic domain 45:90 human chitotriosidase (CHIT1) catalytic domain 45:90 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 7 11 theme pseudo-apo 1059:1068 arg1 forms 1070:1074 the apo and pseudo-apo forms 1047:1074 forms 1070:1074 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 7 12 with complex 1084:1090 arg1 chitobiose 1097:1106 chitobiose 1097:1106 chitobiose 1097:1106 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 9 13 theme catalytic 1389:1397 arg1 triad 1399:1403 the catalytic triad 1385:1403 the catalytic triad 1385:1403 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 0 14 theme atomic 95:100 arg1 diffraction 119:129 atomic resolution X-ray diffraction 95:129 atomic resolution X-ray diffraction 95:129 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 3 15 theme immune-system 452:464 arg1 disorders 466:474 immune-system disorders 452:474 immune-system disorders 452:474 Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. 26143917 9 16 theme transglycosylation 1486:1503 arg1 activities 1505:1514 the hydrolysis and transglycosylation activities 1467:1514 activities 1505:1514 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 4 17 theme hydrolysis 635:644 arg1 reaction 646:653 the hydrolysis reaction 631:653 the hydrolysis reaction 631:653 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 9 18 theme triad 1399:1403 arg1 conformation 1347:1358 conformation 1347:1358 conformation 1347:1358 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 9 18 theme triad 1399:1403 arg1 state 1376:1380 protonation state 1364:1380 protonation state 1364:1380 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 5 19 theme other 710:714 arg1 homologues 716:725 other homologues 710:725 other homologues 710:725 Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. 26143917 7 20 theme apo 1051:1053 arg1 forms 1070:1074 the apo and pseudo-apo forms 1047:1074 forms 1070:1074 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 5 21 theme additional 738:747 arg1 transglycosylation 759:776 transglycosylation 759:776 transglycosylation 759:776 Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. 26143917 5 21 theme additional 738:747 arg1 activity 749:756 an additional activity 735:756 an additional activity 735:756 Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. 26143917 7 22 theme PDB 1012:1014 arg1 1waw 1022:1025 PDB entry 1waw 1012:1025 PDB entry 1waw 1012:1025 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 7 22 theme PDB 1012:1014 arg1 1.65 Å 1004:1009 1.65 Å 1004:1009 1.65 Å 1004:1009 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 2 23 theme structural 362:371 arg1 purposes 373:380 structural purposes 362:380 structural purposes 362:380 GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. 26143917 0 24 theme New 0:2 arg1 insights 4:11 New insights 0:11 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 0:147 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 7 25 theme catalytic 969:977 arg1 domain 979:984 the catalytic domain 965:984 the catalytic domain 965:984 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 9 26 theme protonation 1364:1374 arg1 state 1376:1380 protonation state 1364:1380 protonation state 1364:1380 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 0 27 theme X-ray 113:117 arg1 diffraction 119:129 atomic resolution X-ray diffraction 95:129 atomic resolution X-ray diffraction 95:129 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 7 28 dep 1.65 Å 1004:1009 arg1 0.95-1.10 Å 1031:1041 0.95-1.10 Å 1031:1041 0.95-1.10 Å 1031:1041 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 7 28 dep 1.65 Å 1004:1009 arg1 to 1028:1029 to 1028:1029 to 1028:1029 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 4 29 theme conserved 548:556 arg1 cluster 558:564 a conserved cluster 546:564 a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction 546:653 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 0 30 theme resolution 102:111 arg1 diffraction 119:129 atomic resolution X-ray diffraction 95:129 atomic resolution X-ray diffraction 95:129 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 6 31 theme enzymatic 849:857 arg1 activity 859:866 the dual enzymatic activity 840:866 the dual enzymatic activity 840:866 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 2 32 theme GH18 265:268 arg1 enzymes 270:276 GH18 enzymes 265:276 GH18 enzymes 265:276 GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. 26143917 1 33 theme human 178:182 arg1 chitinase 184:192 a human chitinase 176:192 a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18) 176:262 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 1 33 theme human 178:182 arg1 Chitotriosidase 149:163 Chitotriosidase 149:163 Chitotriosidase (CHIT1) 149:171 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 8 34 theme mechanics/molecular 1234:1252 arg1 calculations 1272:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. 26143917 7 35 theme entry 1016:1020 arg1 1waw 1022:1025 PDB entry 1waw 1012:1025 PDB entry 1waw 1012:1025 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 7 35 theme entry 1016:1020 arg1 1.65 Å 1004:1009 1.65 Å 1004:1009 1.65 Å 1004:1009 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 6 36 theme dual 844:847 arg1 activity 859:866 the dual enzymatic activity 840:866 the dual enzymatic activity 840:866 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 8 37 theme quantum 1226:1232 arg1 calculations 1272:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. 26143917 0 38 theme enzymatic 22:30 arg1 mechanism 32:40 the enzymatic mechanism 18:40 the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM 18:146 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 5 39 theme substrate 689:697 arg1 concentration 672:684 an excess concentration 662:684 an excess concentration of substrate 662:697 Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. 26143917 8 40 theme hybrid 1219:1224 arg1 calculations 1272:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. 26143917 0 41 theme human 45:49 arg1 domain 85:90 human chitotriosidase (CHIT1) catalytic domain 45:90 human chitotriosidase (CHIT1) catalytic domain 45:90 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 7 42 theme protonation 1143:1153 arg1 states 1155:1160 the protonation states 1139:1160 the protonation states within the active site 1139:1183 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 9 43 dep conformation 1347:1358 arg1 the 1343:1345 the 1343:1345 the 1343:1345 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 9 44 theme new 1419:1421 arg1 role 1423:1426 a new role 1417:1426 a new role for Tyr27 1417:1436 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 3 45 from upregulation 436:447 arg1 disorders 466:474 immune-system disorders 452:474 immune-system disorders 452:474 Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. 26143917 8 46 theme mechanics 1254:1262 arg1 calculations 1272:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. 26143917 4 47 theme catalytic 523:531 arg1 domain 533:538 The 39 kDa catalytic domain 512:538 The 39 kDa catalytic domain 512:538 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 0 48 theme hybrid 135:140 arg1 QM/MM 142:146 hybrid QM/MM 135:146 hybrid QM/MM 135:146 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 6 49 theme chitinases 825:834 arg1 mechanism 807:815 the catalytic mechanism 793:815 the catalytic mechanism of GH18 chitinases 793:834 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 6 49 theme chitinases 825:834 arg1 activity 859:866 the dual enzymatic activity 840:866 the dual enzymatic activity 840:866 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 7 50 theme states 1155:1160 arg1 determination 1122:1134 the determination 1118:1134 the determination of the protonation states within the active site 1118:1183 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 2 51 theme N-acetylglucosamine 299:317 arg1 polymer 319:325 an N-acetylglucosamine polymer 296:325 an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes 296:380 GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. 26143917 2 51 theme N-acetylglucosamine 299:317 arg1 chitin 288:293 chitin 288:293 chitin 288:293 GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. 26143917 4 52 theme 39 kDa 516:521 arg1 domain 533:538 The 39 kDa catalytic domain 512:538 The 39 kDa catalytic domain 512:538 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 6 53 theme GH18 820:823 arg1 chitinases 825:834 GH18 chitinases 820:834 GH18 chitinases 820:834 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 9 54 theme new 1308:1310 arg1 mechanism 1312:1320 a new mechanism 1306:1320 a new mechanism involving changes in the conformation and protonation state of the catalytic triad 1306:1403 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 7 55 theme domain 979:984 arg1 crystals 953:960 the crystals 949:960 the crystals of the catalytic domain 949:984 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 8 56 theme QM/MM 1265:1269 arg1 calculations 1272:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 hybrid quantum mechanics/molecular mechanics (QM/MM) calculations 1219:1283 This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. 26143917 2 57 theme lower 342:346 arg1 organisms 348:356 lower organisms 342:356 lower organisms for structural purposes 342:380 GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. 26143917 4 58 theme residues 582:589 arg1 cluster 558:564 a conserved cluster 546:564 a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction 546:653 The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. 26143917 7 59 theme active 1173:1178 arg1 site 1180:1183 the active site 1169:1183 the active site 1169:1183 The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. 26143917 9 60 theme new 1449:1451 arg1 insights 1453:1460 new insights 1449:1460 new insights into the hydrolysis and transglycosylation activities 1449:1514 The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities. 26143917 3 61 theme Gaucher 495:501 arg1 disease 503:509 Gaucher disease 495:509 Gaucher disease 495:509 Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. 26143917 6 62 dep structure 873:881 arg1 the 869:871 the 869:871 the 869:871 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 0 63 theme chitotriosidase 51:65 arg1 domain 85:90 human chitotriosidase (CHIT1) catalytic domain 45:90 human chitotriosidase (CHIT1) catalytic domain 45:90 New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM. 26143917 6 64 theme CHIT1 900:904 arg1 mechanism 887:895 mechanism 887:895 mechanism 887:895 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 6 64 theme CHIT1 900:904 arg1 structure 873:881 structure 873:881 structure 873:881 To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. 26143917 5 65 theme excess 665:670 arg1 concentration 672:684 an excess concentration 662:684 an excess concentration of substrate 662:697 Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. 26143917 3 66 theme disease 503:509 arg1 marker 485:490 a marker 483:490 a marker of Gaucher disease 483:509 Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. 26143917 1 67 theme conserved 218:226 arg1 family 247:252 the highly conserved glycosyl hydrolase family 18 207:255 the highly conserved glycosyl hydrolase family 18 (GH18) 207:262 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 26143917 1 67 theme conserved 218:226 arg1 GH18 258:261 GH18 258:261 GH18 258:261 Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). 24909990 7 0 theme pentameric 964:973 arg1 channels 992:999 pentameric ligand-gated ion channels 964:999 pentameric ligand-gated ion channels 964:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 1 1 theme synaptic 140:147 arg1 transmission 149:160 rapid inhibitory synaptic transmission 123:160 rapid inhibitory synaptic transmission 123:160 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 2 theme unexpected 584:593 arg1 role 606:609 an unexpected structural role 581:609 an unexpected structural role for a conserved N-linked glycan 581:641 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 4 3 theme mechanistic 510:520 arg1 consequences 522:533 the mechanistic consequences 506:533 the mechanistic consequences of multiple human disease mutations 506:569 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 7 4 theme signalling 939:948 arg1 mechanisms 950:959 the signalling mechanisms 935:959 the signalling mechanisms of pentameric ligand-gated ion channels 935:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 1 5 theme transmission 149:160 arg1 mediators 110:118 the principal mediators 96:118 the principal mediators of rapid inhibitory synaptic transmission in the human brain 96:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 5 theme transmission 149:160 arg1 receptors 72:80 Type-A γ-aminobutyric acid receptors 45:80 Type-A γ-aminobutyric acid receptors (GABAARs) 45:90 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 6 theme architectural 433:445 arg1 elements 447:454 architectural elements 433:454 architectural elements unique to eukaryotic Cys-loop receptors 433:494 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 6 7 theme desensitized 877:888 arg1 state 890:894 a desensitized state 875:894 a desensitized state 875:894 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 1 8 theme Type-A 45:50 arg1 mediators 110:118 the principal mediators 96:118 the principal mediators of rapid inhibitory synaptic transmission in the human brain 96:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 8 theme Type-A 45:50 arg1 GABAARs 83:89 GABAARs 83:89 GABAARs 83:89 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 8 theme Type-A 45:50 arg1 receptors 72:80 Type-A γ-aminobutyric acid receptors 45:80 Type-A γ-aminobutyric acid receptors (GABAARs) 45:90 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 3 9 theme GABAAR 356:361 arg1 structure 341:349 the first three-dimensional structure 313:349 the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution 313:407 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 4 10 theme eukaryotic 466:475 arg1 receptors 486:494 eukaryotic Cys-loop receptors 466:494 eukaryotic Cys-loop receptors 466:494 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 1 11 theme γ-aminobutyric 52:65 arg1 mediators 110:118 the principal mediators 96:118 the principal mediators of rapid inhibitory synaptic transmission in the human brain 96:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 11 theme γ-aminobutyric 52:65 arg1 GABAARs 83:89 GABAARs 83:89 GABAARs 83:89 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 11 theme γ-aminobutyric 52:65 arg1 receptors 72:80 Type-A γ-aminobutyric acid receptors 45:80 Type-A γ-aminobutyric acid receptors (GABAARs) 45:90 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 12 theme structural 595:604 arg1 role 606:609 an unexpected structural role 581:609 an unexpected structural role for a conserved N-linked glycan 581:641 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 6 13 theme channel 797:803 arg1 region 805:810 The channel region 793:810 The channel region 793:810 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 1 14 theme acid 67:70 arg1 mediators 110:118 the principal mediators 96:118 the principal mediators of rapid inhibitory synaptic transmission in the human brain 96:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 14 theme acid 67:70 arg1 GABAARs 83:89 GABAARs 83:89 GABAARs 83:89 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 14 theme acid 67:70 arg1 receptors 72:80 Type-A γ-aminobutyric acid receptors 45:80 Type-A γ-aminobutyric acid receptors (GABAARs) 45:90 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 5 15 theme modulators 781:790 arg1 design 764:769 the rational design 751:769 the rational design of GABAAR modulators 751:790 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 2 16 from decline 184:190 arg1 signalling 202:211 GABAAR signalling 195:211 GABAAR signalling 195:211 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 5 17 theme GABAAR 774:779 arg1 modulators 781:790 GABAAR modulators 774:790 GABAAR modulators 774:790 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 1 18 from mediators 110:118 arg1 brain 175:179 the human brain 165:179 the human brain 165:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 0 19 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a human GABAA receptor 0:42 Crystal structure of a human GABAA receptor. 24909990 3 20 theme human 368:372 arg1 homopentamer 377:388 the human β3 homopentamer 364:388 the human β3 homopentamer 364:388 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 3 20 theme human 368:372 arg1 GABAAR 356:361 a GABAAR 354:361 a GABAAR 354:361 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 4 21 theme Cys-loop 477:484 arg1 receptors 486:494 eukaryotic Cys-loop receptors 466:494 eukaryotic Cys-loop receptors 466:494 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 1 22 theme human 169:173 arg1 brain 175:179 the human brain 165:179 the human brain 165:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 3 23 theme β3 374:375 arg1 homopentamer 377:388 the human β3 homopentamer 364:388 the human β3 homopentamer 364:388 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 3 23 theme β3 374:375 arg1 GABAAR 356:361 a GABAAR 354:361 a GABAAR 354:361 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 4 24 theme mutations 561:569 arg1 consequences 522:533 the mechanistic consequences 506:533 the mechanistic consequences of multiple human disease mutations 506:569 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 6 25 from base 839:842 arg1 gate 827:830 a closed gate 818:830 a closed gate at the base of the pore 818:854 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 6 25 from base 839:842 arg1 representative 857:870 representative 857:870 representative of a desensitized state 857:894 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 4 26 theme disease 553:559 arg1 mutations 561:569 multiple human disease mutations 538:569 multiple human disease mutations 538:569 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 5 27 dep crystallized 661:672 arg1 bound 674:678 bound 674:678 bound 674:678 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 4 28 link N-linked 627:634 arg1 glycan 636:641 a conserved N-linked glycan 615:641 a conserved N-linked glycan 615:641 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 0 29 theme GABAA 29:33 arg1 receptor 35:42 a human GABAA receptor 21:42 a human GABAA receptor 21:42 Crystal structure of a human GABAA receptor. 24909990 2 30 theme hyperactive 222:232 arg1 anxiety 275:281 anxiety 275:281 anxiety 275:281 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 30 theme hyperactive 222:232 arg1 disorders 247:255 hyperactive neurological disorders 222:255 hyperactive neurological disorders such as insomnia, anxiety and epilepsy 222:294 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 30 theme hyperactive 222:232 arg1 insomnia 265:272 insomnia 265:272 insomnia 265:272 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 30 theme hyperactive 222:232 arg1 epilepsy 287:294 epilepsy 287:294 epilepsy 287:294 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 0 31 theme human 23:27 arg1 receptor 35:42 a human GABAA receptor 21:42 a human GABAA receptor 21:42 Crystal structure of a human GABAA receptor. 24909990 3 32 theme 3 Å 394:396 arg1 resolution 398:407 3 Å resolution 394:407 3 Å resolution 394:407 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 4 33 theme unique 456:461 arg1 elements 447:454 architectural elements 433:454 architectural elements unique to eukaryotic Cys-loop receptors 433:494 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 7 34 theme ligand-gated 975:986 arg1 channels 992:999 pentameric ligand-gated ion channels 964:999 pentameric ligand-gated ion channels 964:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 7 35 theme GABAergic 1038:1046 arg1 neurotransmission 1048:1064 GABAergic neurotransmission 1038:1064 GABAergic neurotransmission 1038:1064 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 5 36 theme rational 755:762 arg1 design 764:769 the rational design 751:769 the rational design of GABAAR modulators 751:790 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 6 37 theme pore 851:854 arg1 base 839:842 the base 835:842 the base of the pore 835:854 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 7 38 theme current 1013:1019 arg1 understanding 1021:1033 current understanding 1013:1033 current understanding of GABAergic neurotransmission 1013:1064 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 0 39 theme receptor 35:42 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a human GABAA receptor 0:42 Crystal structure of a human GABAA receptor. 24909990 4 40 theme human 547:551 arg1 mutations 561:569 multiple human disease mutations 538:569 multiple human disease mutations 538:569 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 7 41 theme neurotransmission 1048:1064 arg1 understanding 1021:1033 current understanding 1013:1033 current understanding of GABAergic neurotransmission 1013:1064 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 6 42 theme state 890:894 arg1 representative 857:870 representative 857:870 representative of a desensitized state 857:894 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 6 42 theme state 890:894 arg1 gate 827:830 a closed gate 818:830 a closed gate at the base of the pore 818:854 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 2 43 theme GABAAR 195:200 arg1 signalling 202:211 GABAAR signalling 195:211 GABAAR signalling 195:211 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 7 44 theme channels 992:999 arg1 mechanisms 950:959 the signalling mechanisms 935:959 the signalling mechanisms of pentameric ligand-gated ion channels 935:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 1 45 theme principal 100:108 arg1 mediators 110:118 the principal mediators 96:118 the principal mediators of rapid inhibitory synaptic transmission in the human brain 96:179 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 1 45 theme principal 100:108 arg1 receptors 72:80 Type-A γ-aminobutyric acid receptors 45:80 Type-A γ-aminobutyric acid receptors (GABAARs) 45:90 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 46 theme multiple 538:545 arg1 mutations 561:569 multiple human disease mutations 538:569 multiple human disease mutations 538:569 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 5 47 theme unknown 696:702 arg1 benzamidine 713:723 benzamidine 713:723 benzamidine 713:723 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 5 47 theme unknown 696:702 arg1 agonist 704:710 a previously unknown agonist 683:710 a previously unknown agonist 683:710 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 7 48 theme new 917:919 arg1 insights 921:928 new insights 917:928 new insights into the signalling mechanisms of pentameric ligand-gated ion channels 917:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 3 49 from resolution 398:407 arg1 structure 341:349 the first three-dimensional structure 313:349 the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution 313:407 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 5 50 theme new 736:738 arg1 avenue 740:745 a new avenue 734:745 a new avenue for the rational design of GABAAR modulators 734:790 The receptor was crystallized bound to a previously unknown agonist, benzamidine, opening a new avenue for the rational design of GABAAR modulators. 24909990 3 51 theme first 317:321 arg1 structure 341:349 the first three-dimensional structure 313:349 the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution 313:407 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 6 52 theme closed 820:825 arg1 gate 827:830 a closed gate 818:830 a closed gate at the base of the pore 818:854 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 6 52 theme closed 820:825 arg1 representative 857:870 representative 857:870 representative of a desensitized state 857:894 The channel region forms a closed gate at the base of the pore, representative of a desensitized state. 24909990 7 53 theme ion 988:990 arg1 channels 992:999 pentameric ligand-gated ion channels 964:999 pentameric ligand-gated ion channels 964:999 These results offer new insights into the signalling mechanisms of pentameric ligand-gated ion channels and enhance current understanding of GABAergic neurotransmission. 24909990 1 54 theme rapid 123:127 arg1 transmission 149:160 rapid inhibitory synaptic transmission 123:160 rapid inhibitory synaptic transmission 123:160 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 55 theme N-linked 627:634 arg1 glycan 636:641 a conserved N-linked glycan 615:641 a conserved N-linked glycan 615:641 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 24909990 3 56 theme three-dimensional 323:339 arg1 structure 341:349 the first three-dimensional structure 313:349 the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution 313:407 Here we present the first three-dimensional structure of a GABAAR, the human β3 homopentamer, at 3 Å resolution. 24909990 2 57 theme neurological 234:245 arg1 anxiety 275:281 anxiety 275:281 anxiety 275:281 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 57 theme neurological 234:245 arg1 disorders 247:255 hyperactive neurological disorders 222:255 hyperactive neurological disorders such as insomnia, anxiety and epilepsy 222:294 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 57 theme neurological 234:245 arg1 insomnia 265:272 insomnia 265:272 insomnia 265:272 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 2 57 theme neurological 234:245 arg1 epilepsy 287:294 epilepsy 287:294 epilepsy 287:294 A decline in GABAAR signalling triggers hyperactive neurological disorders such as insomnia, anxiety and epilepsy. 24909990 1 58 theme inhibitory 129:138 arg1 transmission 149:160 rapid inhibitory synaptic transmission 123:160 rapid inhibitory synaptic transmission 123:160 Type-A γ-aminobutyric acid receptors (GABAARs) are the principal mediators of rapid inhibitory synaptic transmission in the human brain. 24909990 4 59 theme conserved 617:625 arg1 glycan 636:641 a conserved N-linked glycan 615:641 a conserved N-linked glycan 615:641 This structure reveals architectural elements unique to eukaryotic Cys-loop receptors, explains the mechanistic consequences of multiple human disease mutations and shows an unexpected structural role for a conserved N-linked glycan. 27722599 5 0 theme gastric 794:800 arg1 patients 809:816 gastric cancer patients 794:816 gastric cancer patients 794:816 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 10 1 theme gastric 1570:1576 arg1 cancer 1578:1583 gastric cancer 1570:1583 gastric cancer 1570:1583 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 6 2 theme healthy 902:908 arg1 controls 910:917 30 healthy controls 899:917 30 healthy controls 899:917 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 1 3 theme cancer 137:142 arg1 rates 154:158 the highest cancer mortality rates 125:158 the highest cancer mortality rates 125:158 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 0 4 theme nano 74:77 arg1 LC/MS 79:83 nano LC/MS 74:83 nano LC/MS 74:83 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 1 5 contain has 114:116 arg1 cancer 107:112 Gastric cancer 99:112 Gastric cancer 99:112 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 1 5 contain has 114:116 arg2 worldwide 160:168 worldwide 160:168 worldwide 160:168 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 10 6 theme discovery 1556:1564 arg1 purposes 1534:1541 the purposes 1530:1541 the purposes of biomarker discovery for gastric cancer 1530:1583 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 7 7 theme based 986:990 arg1 analysis 1004:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis was performed to identify potential glyco-markers for gastric cancer. 27722599 1 8 theme mortality 144:152 arg1 rates 154:158 the highest cancer mortality rates 125:158 the highest cancer mortality rates 125:158 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 7 9 theme statistical 992:1002 arg1 analysis 1004:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis was performed to identify potential glyco-markers for gastric cancer. 27722599 3 10 theme Serum-based 361:371 arg1 profiling 387:395 Serum-based global glycan profiling 361:395 Serum-based global glycan profiling using mass spectrometry 361:419 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 9 11 theme fucosylated 1276:1286 arg1 N-glycans 1301:1309 the fucosylated complex type N-glycans 1272:1309 the fucosylated complex type N-glycans 1272:1309 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 2 12 theme cancer 353:358 arg1 types 344:348 various types 336:348 various types of cancer 336:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 12 theme cancer 353:358 arg1 inflammation 319:330 inflammation 319:330 inflammation 319:330 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 6 13 theme sex-matched 846:856 arg1 samples 867:873 sex-matched 60 serum samples 846:873 sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) 846:918 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 2 14 theme many 289:292 arg1 types 344:348 various types 336:348 various types of cancer 336:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 14 theme many 289:292 arg1 diseases 300:307 many human diseases 289:307 many human diseases including inflammation and various types of cancer 289:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 14 theme many 289:292 arg1 inflammation 319:330 inflammation 319:330 inflammation 319:330 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 15 theme human 294:298 arg1 types 344:348 various types 336:348 various types of cancer 336:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 15 theme human 294:298 arg1 diseases 300:307 many human diseases 289:307 many human diseases including inflammation and various types of cancer 289:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 2 15 theme human 294:298 arg1 inflammation 319:330 inflammation 319:330 inflammation 319:330 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 3 16 theme global 373:378 arg1 profiling 387:395 Serum-based global glycan profiling 361:395 Serum-based global glycan profiling using mass spectrometry 361:419 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 5 17 theme protein-specific 676:691 arg1 purification 708:719 protein-specific immunoaffinity purification 676:719 protein-specific immunoaffinity purification 676:719 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 5 18 gly glycosylation 777:789 arg1 patients 809:816 gastric cancer patients 794:816 gastric cancer patients 794:816 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 10 19 dep quantitative 1454:1465 arg1 structure-specific 1468:1485 structure-specific 1468:1485 structure-specific 1468:1485 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 5 20 theme cancer 802:807 arg1 patients 809:816 gastric cancer patients 794:816 gastric cancer patients 794:816 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 4 21 theme clinical 644:651 arg1 use 653:655 clinical use 644:655 clinical use 644:655 However, localization of the aberrant glycosylation is desirable in order to improve the specificity and sensitivity for clinical use. 27722599 1 22 theme rates 154:158 arg1 one 118:120 one 118:120 one 118:120 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 1 22 theme rates 154:158 arg1 rates 154:158 the highest cancer mortality rates 125:158 the highest cancer mortality rates 125:158 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 5 23 theme haptoglobin 765:775 arg1 glycosylation 777:789 haptoglobin glycosylation 765:789 haptoglobin glycosylation of gastric cancer patients 765:816 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 5 24 theme patients 809:816 arg1 glycosylation 777:789 haptoglobin glycosylation 765:789 haptoglobin glycosylation of gastric cancer patients 765:816 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 3 25 theme mass 403:406 arg1 spectrometry 408:419 mass spectrometry 403:419 mass spectrometry 403:419 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 6 26 theme haptoglobin 954:964 arg1 N-glycans 966:974 haptoglobin N-glycans 954:974 haptoglobin N-glycans 954:974 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 3 27 theme glycan 380:385 arg1 profiling 387:395 Serum-based global glycan profiling 361:395 Serum-based global glycan profiling using mass spectrometry 361:419 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 8 28 theme several 1112:1118 arg1 N-glycans 1157:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 9 29 theme type 1410:1413 arg1 fucose 1349:1354 antenna fucose 1341:1354 antenna fucose 1341:1354 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 9 29 theme type 1410:1413 arg1 structures 1415:1424 sialyl-Lex type structures 1399:1424 sialyl-Lex type structures 1399:1424 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 8 30 gly fucosylated 1145:1155 arg1 N-glycans 1157:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 0 31 theme Glycomic 0:7 arg1 profiling 9:17 Glycomic profiling 0:17 Glycomic profiling of targeted serum haptoglobin for gastric cancer 0:66 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 6 32 dep samples 867:873 arg1 patients 886:893 30 cancer patients 876:893 30 cancer patients 876:893 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 6 32 dep samples 867:873 arg1 controls 910:917 30 healthy controls 899:917 30 healthy controls 899:917 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 6 33 theme serum 861:865 arg1 samples 867:873 sex-matched 60 serum samples 846:873 sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) 846:918 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 9 34 theme complex 1288:1294 arg1 N-glycans 1301:1309 the fucosylated complex type N-glycans 1272:1309 the fucosylated complex type N-glycans 1272:1309 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 2 35 gly glycosylation 238:250 arg1 proteins 261:268 serum proteins 255:268 serum proteins 255:268 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 4 36 theme glycosylation 561:573 arg1 localization 532:543 localization 532:543 localization of the aberrant glycosylation 532:573 However, localization of the aberrant glycosylation is desirable in order to improve the specificity and sensitivity for clinical use. 27722599 10 37 theme biomarker 1546:1554 arg1 discovery 1556:1564 biomarker discovery 1546:1564 biomarker discovery for gastric cancer 1546:1583 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 7 38 theme T-test 979:984 arg1 analysis 1004:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis 977:1011 A T-test based statistical analysis was performed to identify potential glyco-markers for gastric cancer. 27722599 2 39 from glycosylation 238:250 arg1 proteins 261:268 serum proteins 255:268 serum proteins 255:268 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 3 40 theme potential 470:478 arg1 markers 487:493 several potential glycan markers 462:493 several potential glycan markers for several disease states 462:520 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 8 41 theme tri- 1120:1123 arg1 N-glycans 1157:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 10 42 theme haptoglobin 1500:1510 arg1 glycosylation 1512:1524 haptoglobin glycosylation 1500:1524 haptoglobin glycosylation 1500:1524 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 1 43 theme Gastric 99:105 arg1 cancer 107:112 Gastric cancer 99:112 Gastric cancer 99:112 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 3 44 theme glycan 480:485 arg1 markers 487:493 several potential glycan markers 462:493 several potential glycan markers for several disease states 462:520 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 0 45 theme haptoglobin 37:47 arg1 profiling 9:17 Glycomic profiling 0:17 Glycomic profiling of targeted serum haptoglobin for gastric cancer 0:66 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 8 46 theme fucosylated 1145:1155 arg1 N-glycans 1157:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 2 47 theme Aberrant 229:236 arg1 glycosylation 238:250 Aberrant glycosylation 229:250 Aberrant glycosylation in serum proteins 229:268 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 0 48 theme serum 31:35 arg1 haptoglobin 37:47 targeted serum haptoglobin 22:47 targeted serum haptoglobin for gastric cancer 22:66 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 10 49 theme glycosylation 1512:1524 arg1 profiling 1487:1495 quantitative, structure-specific profiling 1454:1495 quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer 1454:1583 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 5 50 theme MS 742:743 arg1 analysis 745:752 MS analysis 742:752 MS analysis 742:752 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 0 51 theme targeted 22:29 arg1 haptoglobin 37:47 targeted serum haptoglobin 22:47 targeted serum haptoglobin for gastric cancer 22:66 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 1 52 from difficulties 190:201 arg1 detection 218:226 early-stage detection 206:226 early-stage detection 206:226 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 3 53 theme several 462:468 arg1 markers 487:493 several potential glycan markers 462:493 several potential glycan markers for several disease states 462:520 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 0 54 theme gastric 53:59 arg1 cancer 61:66 gastric cancer 53:66 gastric cancer 53:66 Glycomic profiling of targeted serum haptoglobin for gastric cancer using nano LC/MS and LC/MS/MS. 27722599 6 55 used used 925:928 arg2 Age- 837:840 Age- 837:840 Age- 837:840 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 6 55 used used 925:928 arg2 samples 867:873 sex-matched 60 serum samples 846:873 sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) 846:918 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 5 56 theme immunoaffinity 693:706 arg1 purification 708:719 protein-specific immunoaffinity purification 676:719 protein-specific immunoaffinity purification 676:719 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 6 57 theme cancer 879:884 arg1 patients 886:893 30 cancer patients 876:893 30 cancer patients 876:893 Age- and sex-matched 60 serum samples (30 cancer patients and 30 healthy controls) were used to profile and quantify haptoglobin N-glycans. 27722599 3 58 theme several 499:505 arg1 states 515:520 several disease states 499:520 several disease states 499:520 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 8 59 theme tetra-antennary 1129:1143 arg1 N-glycans 1157:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 several tri- and tetra-antennary fucosylated N-glycans 1112:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 4 60 theme aberrant 552:559 arg1 glycosylation 561:573 the aberrant glycosylation 548:573 the aberrant glycosylation 548:573 However, localization of the aberrant glycosylation is desirable in order to improve the specificity and sensitivity for clinical use. 27722599 8 61 theme N-glycans 1157:1165 arg1 abundances 1098:1107 abundances 1098:1107 abundances of several tri- and tetra-antennary fucosylated N-glycans 1098:1165 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 3 62 theme disease 507:513 arg1 states 515:520 several disease states 499:520 several disease states 499:520 Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. 27722599 9 63 theme type 1296:1299 arg1 N-glycans 1301:1309 the fucosylated complex type N-glycans 1272:1309 the fucosylated complex type N-glycans 1272:1309 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 7 64 theme gastric 1067:1073 arg1 cancer 1075:1080 gastric cancer 1067:1080 gastric cancer 1067:1080 A T-test based statistical analysis was performed to identify potential glyco-markers for gastric cancer. 27722599 9 65 theme antenna 1341:1347 arg1 sialyl-Lea 1385:1394 sialyl-Lea 1385:1394 sialyl-Lea 1385:1394 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 9 65 theme antenna 1341:1347 arg1 fucose 1349:1354 antenna fucose 1341:1354 antenna fucose 1341:1354 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 9 65 theme antenna 1341:1347 arg1 structures 1415:1424 sialyl-Lex type structures 1399:1424 sialyl-Lex type structures 1399:1424 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 7 66 theme potential 1039:1047 arg1 glyco-markers 1049:1061 potential glyco-markers 1039:1061 potential glyco-markers for gastric cancer 1039:1080 A T-test based statistical analysis was performed to identify potential glyco-markers for gastric cancer. 27722599 2 67 theme various 336:342 arg1 types 344:348 various types 336:348 various types of cancer 336:358 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 1 68 theme early-stage 206:216 arg1 detection 218:226 early-stage detection 206:226 early-stage detection 206:226 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27722599 5 69 theme glycan 722:727 arg1 release 729:735 glycan release 722:735 glycan release 722:735 Here, we combined protein-specific immunoaffinity purification, glycan release, and MS analysis to examine haptoglobin glycosylation of gastric cancer patients for glyco-markers. 27722599 9 70 gly fucosylated 1276:1286 arg1 N-glycans 1301:1309 the fucosylated complex type N-glycans 1272:1309 the fucosylated complex type N-glycans 1272:1309 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 2 71 theme serum 255:259 arg1 proteins 261:268 serum proteins 255:268 serum proteins 255:268 Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. 27722599 9 72 theme sialyl-Lex 1399:1408 arg1 fucose 1349:1354 antenna fucose 1341:1354 antenna fucose 1341:1354 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 9 72 theme sialyl-Lex 1399:1408 arg1 structures 1415:1424 sialyl-Lex type structures 1399:1424 sialyl-Lex type structures 1399:1424 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 8 73 theme cancer 1193:1198 arg1 patients 1200:1207 gastric cancer patients 1185:1207 gastric cancer patients 1185:1207 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 4 74 dep specificity 612:622 arg1 the 608:610 the 608:610 the 608:610 However, localization of the aberrant glycosylation is desirable in order to improve the specificity and sensitivity for clinical use. 27722599 10 75 theme quantitative 1454:1465 arg1 profiling 1487:1495 quantitative, structure-specific profiling 1454:1495 quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer 1454:1583 This platform demonstrates quantitative, structure-specific profiling of haptoglobin glycosylation for the purposes of biomarker discovery for gastric cancer. 27722599 8 76 theme gastric 1185:1191 arg1 patients 1200:1207 gastric cancer patients 1185:1207 gastric cancer patients 1185:1207 Interestingly, abundances of several tri- and tetra-antennary fucosylated N-glycans were increased in gastric cancer patients. 27722599 9 77 theme structural 1224:1233 arg1 analysis 1235:1242 structural analysis 1224:1242 structural analysis via LC/MS/MS 1224:1255 Additionally, structural analysis via LC/MS/MS indicated that the fucosylated complex type N-glycans were primarily decorated with antenna fucose, which can be categorized as sialyl-Lea or sialyl-Lex type structures. 27722599 1 78 theme highest 129:135 arg1 rates 154:158 the highest cancer mortality rates 125:158 the highest cancer mortality rates 125:158 Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. 27574189 3 0 theme Purified 264:271 arg1 ADAMTS13 273:280 Purified ADAMTS13 264:280 Purified ADAMTS13 264:280 Purified ADAMTS13 was reduced, alkylated, and processed into peptides with either trypsin or chymotrypsin. 27574189 13 1 theme disintegrin 1317:1327 arg1 domain 1329:1334 the disintegrin domain 1313:1334 the disintegrin domain 1313:1334 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 15 2 theme C-mannosylation 1419:1433 arg1 sites 1435:1439 C-mannosylation sites 1419:1439 C-mannosylation sites 1419:1439 C-mannosylation sites were identified in TSP1, linker TSP4-TSP5, and TSP8. 27574189 16 3 theme glycan 1544:1549 arg1 modifications 1551:1563 glycan modifications 1544:1563 glycan modifications 1544:1563 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 13 4 theme O-fucosylation 1280:1293 arg1 site 1295:1298 one additional O-fucosylation site 1265:1298 one additional O-fucosylation site 1265:1298 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 7 5 theme mannose-containing 798:815 arg1 glycan 817:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 27574189 4 6 theme mass 456:459 arg1 spectrometry 461:472 tandem mass spectrometry 449:472 tandem mass spectrometry employing higher-energy collision dissociation fragmentation 449:533 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 5 7 theme glycan 576:581 arg1 fragment 583:590 a glycan fragment 574:590 a glycan fragment 574:590 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 11 8 theme repeats 1161:1167 arg1 modification 1123:1134 a common posttranslational modification 1096:1134 a common posttranslational modification of thrombospondin type 1 repeats 1096:1167 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 11 8 theme repeats 1161:1167 arg1 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 4 9 theme HILIC 418:422 arg1 zip-tips 424:431 zwitterionic HILIC zip-tips 405:431 zwitterionic HILIC zip-tips 405:431 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 11 10 theme type 1154:1157 arg1 repeats 1161:1167 thrombospondin type 1 repeats 1139:1167 thrombospondin type 1 repeats 1139:1167 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 0 11 link plasma-derived 29:42 arg1 ADAMTS13 44:51 plasma-derived ADAMTS13 29:51 plasma-derived ADAMTS13 29:51 Identification of glycans on plasma-derived ADAMTS13. 27574189 10 12 contain contained 1044:1052 arg1 glycans 1036:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans contained a terminal sialic acid. 27574189 10 12 contain contained 1044:1052 arg2 acid 1072:1075 a terminal sialic acid 1054:1075 a terminal sialic acid 1054:1075 All 6 O-linked glycans contained a terminal sialic acid. 27574189 16 13 from complexity 1530:1539 arg1 ADAMTS13 1568:1575 ADAMTS13 1568:1575 ADAMTS13 1568:1575 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 5 14 theme diagnostic 556:565 arg1 ion 567:569 a diagnostic ion 554:569 a diagnostic ion of a glycan fragment 554:590 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 15 15 theme linker 1466:1471 arg1 TSP4-TSP5 1473:1481 linker TSP4-TSP5 1466:1481 linker TSP4-TSP5 1466:1481 C-mannosylation sites were identified in TSP1, linker TSP4-TSP5, and TSP8. 27574189 1 16 gly glycoprotein 173:184 arg1 glycoprotein 173:184 the plasma glycoprotein ADAMTS13 162:193 the plasma glycoprotein ADAMTS13 162:193 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 2 17 theme ADAMTS13 254:261 arg1 composition 224:234 the glycan composition 213:234 the glycan composition of plasma-derived ADAMTS13 213:261 Here, we studied the glycan composition of plasma-derived ADAMTS13. 27574189 13 18 theme additional 1269:1278 arg1 site 1295:1298 one additional O-fucosylation site 1265:1298 one additional O-fucosylation site 1265:1298 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 1 19 theme plasma 166:171 arg1 ADAMTS13 186:193 the plasma glycoprotein ADAMTS13 162:193 the plasma glycoprotein ADAMTS13 162:193 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 4 20 theme collision 498:506 arg1 fragmentation 521:533 higher-energy collision dissociation fragmentation 484:533 higher-energy collision dissociation fragmentation 484:533 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 16 21 contain containing 1662:1671 arg1 receptors 1652:1660 immune- or clearance receptors 1631:1660 immune- or clearance receptors containing carbohydrate recognition domains 1631:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 16 21 contain containing 1662:1671 arg2 domains 1698:1704 carbohydrate recognition domains 1673:1704 carbohydrate recognition domains 1673:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 16 22 theme recognition 1686:1696 arg1 domains 1698:1704 carbohydrate recognition domains 1673:1704 carbohydrate recognition domains 1673:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 12 23 theme TSP 1230:1232 arg1 repeats 1242:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 We identified 7 O-fucosylation sites in the thrombospondin (TSP) type 1 repeats. 27574189 5 24 theme transfer 602:609 arg1 fragmentation 624:636 electron transfer dissociation fragmentation 593:636 electron transfer dissociation fragmentation 593:636 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 0 25 from glycans 18:24 arg1 ADAMTS13 44:51 plasma-derived ADAMTS13 29:51 plasma-derived ADAMTS13 29:51 Identification of glycans on plasma-derived ADAMTS13. 27574189 8 26 theme O-linked 873:880 arg1 glycans 882:888 Six O-linked glycans 869:888 Six O-linked glycans mostly terminating in sialic acid 869:922 Six O-linked glycans mostly terminating in sialic acid were found dispersed over ADAMTS13. 27574189 9 27 attach attached 987:994 arg1 Thr 1016:1018 one to Thr 1009:1018 one to Thr 1009:1018 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 9 27 attach attached 987:994 arg2 glycans 974:980 Five O-linked glycans 960:980 Five O-linked glycans 960:980 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 9 27 attach attached 987:994 arg1 Ser 1001:1003 a Ser 999:1003 a Ser 999:1003 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 0 28 from Identification 0:13 arg1 ADAMTS13 44:51 plasma-derived ADAMTS13 29:51 plasma-derived ADAMTS13 29:51 Identification of glycans on plasma-derived ADAMTS13. 27574189 6 29 theme N-linked 695:702 arg1 glycans 704:710 N-linked glycans 695:710 N-linked glycans 695:710 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 12 30 theme type 1235:1238 arg1 repeats 1242:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 We identified 7 O-fucosylation sites in the thrombospondin (TSP) type 1 repeats. 27574189 1 31 theme thrombotic 87:96 arg1 purpura 115:121 acquired thrombotic thrombocytopenic purpura 78:121 acquired thrombotic thrombocytopenic purpura 78:121 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 14 32 theme CG 1415:1416 arg1 sequence 1398:1405 the proposed consensus sequence 1375:1405 the proposed consensus sequence CSX(S/T)CG 1375:1416 This O-fucosylation site did not meet the proposed consensus sequence CSX(S/T)CG. 27574189 6 33 theme fucose 774:779 arg1 residues 781:788 fucose residues 774:788 fucose residues 774:788 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 12 34 theme thrombospondin 1214:1227 arg1 repeats 1242:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 the thrombospondin (TSP) type 1 repeats 1210:1248 We identified 7 O-fucosylation sites in the thrombospondin (TSP) type 1 repeats. 27574189 5 35 theme precursor 664:672 arg1 ion 674:676 the same precursor ion 655:676 the same precursor ion 655:676 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 0 36 theme glycans 18:24 arg1 Identification 0:13 Identification 0:13 Identification of glycans on plasma-derived ADAMTS13. 0:52 Identification of glycans on plasma-derived ADAMTS13. 27574189 13 37 located found 1304:1308 arg2 site 1295:1298 one additional O-fucosylation site 1265:1298 one additional O-fucosylation site 1265:1298 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 13 37 located found 1304:1308 arg1 domain 1329:1334 the disintegrin domain 1313:1334 the disintegrin domain 1313:1334 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 15 38 gly C-mannosylation 1419:1433 arg2 sites 1435:1439 C-mannosylation sites 1419:1439 C-mannosylation sites 1419:1439 C-mannosylation sites were identified in TSP1, linker TSP4-TSP5, and TSP8. 27574189 14 39 gly O-fucosylation 1342:1355 arg2 site 1357:1360 This O-fucosylation site 1337:1360 This O-fucosylation site 1337:1360 This O-fucosylation site did not meet the proposed consensus sequence CSX(S/T)CG. 27574189 6 40 theme terminal 748:755 arg1 acids 764:768 terminal sialic acids 748:768 terminal sialic acids 748:768 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 2 41 link plasma-derived 239:252 arg1 ADAMTS13 254:261 plasma-derived ADAMTS13 239:261 plasma-derived ADAMTS13 239:261 Here, we studied the glycan composition of plasma-derived ADAMTS13. 27574189 12 42 theme O-fucosylation 1186:1199 arg1 sites 1201:1205 7 O-fucosylation sites 1184:1205 7 O-fucosylation sites 1184:1205 We identified 7 O-fucosylation sites in the thrombospondin (TSP) type 1 repeats. 27574189 0 43 theme plasma-derived 29:42 arg1 ADAMTS13 44:51 plasma-derived ADAMTS13 29:51 plasma-derived ADAMTS13 29:51 Identification of glycans on plasma-derived ADAMTS13. 27574189 14 44 theme consensus 1388:1396 arg1 sequence 1398:1405 the proposed consensus sequence 1375:1405 the proposed consensus sequence CSX(S/T)CG 1375:1416 This O-fucosylation site did not meet the proposed consensus sequence CSX(S/T)CG. 27574189 6 45 link N-linked 695:702 arg1 glycans 704:710 N-linked glycans 695:710 N-linked glycans 695:710 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 4 46 theme zwitterionic 405:416 arg1 zip-tips 424:431 zwitterionic HILIC zip-tips 405:431 zwitterionic HILIC zip-tips 405:431 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 10 47 link O-linked 1027:1034 arg1 glycans 1036:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans contained a terminal sialic acid. 27574189 9 48 dep Thr 1016:1018 arg1 to 1013:1014 to 1013:1014 to 1013:1014 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 10 49 theme sialic 1065:1070 arg1 acid 1072:1075 a terminal sialic acid 1054:1075 a terminal sialic acid 1054:1075 All 6 O-linked glycans contained a terminal sialic acid. 27574189 2 50 theme glycan 217:222 arg1 composition 224:234 the glycan composition 213:234 the glycan composition of plasma-derived ADAMTS13 213:261 Here, we studied the glycan composition of plasma-derived ADAMTS13. 27574189 13 51 gly O-fucosylation 1280:1293 arg2 site 1295:1298 one additional O-fucosylation site 1265:1298 one additional O-fucosylation site 1265:1298 Unexpectedly, one additional O-fucosylation site was found in the disintegrin domain. 27574189 4 52 theme tandem 449:454 arg1 spectrometry 461:472 tandem mass spectrometry 449:472 tandem mass spectrometry employing higher-energy collision dissociation fragmentation 449:533 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 16 53 theme modifications 1551:1563 arg1 complexity 1530:1539 the complexity 1526:1539 the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains 1526:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 7 54 theme high 793:796 arg1 glycan 817:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 27574189 14 55 theme O-fucosylation 1342:1355 arg1 site 1357:1360 This O-fucosylation site 1337:1360 This O-fucosylation site 1337:1360 This O-fucosylation site did not meet the proposed consensus sequence CSX(S/T)CG. 27574189 5 56 theme fragment 583:590 arg1 ion 567:569 a diagnostic ion 554:569 a diagnostic ion of a glycan fragment 554:590 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 11 57 theme thrombospondin 1139:1152 arg1 repeats 1161:1167 thrombospondin type 1 repeats 1139:1167 thrombospondin type 1 repeats 1139:1167 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 16 58 contain have 1588:1591 arg2 implications 1593:1604 implications 1593:1604 implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains 1593:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 16 58 contain have 1588:1591 arg1 ADAMTS13 1568:1575 ADAMTS13 1568:1575 ADAMTS13 1568:1575 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 11 59 theme posttranslational 1105:1121 arg1 modification 1123:1134 a common posttranslational modification 1096:1134 a common posttranslational modification of thrombospondin type 1 repeats 1096:1167 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 11 59 theme posttranslational 1105:1121 arg1 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 5 60 theme ion 567:569 arg1 detection 541:549 detection 541:549 detection of a diagnostic ion of a glycan fragment 541:590 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 6 61 contain containing 737:746 arg2 residues 781:788 fucose residues 774:788 fucose residues 774:788 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 6 61 contain containing 737:746 arg1 type 732:735 the complex type 720:735 the complex type containing terminal sialic acids and fucose residues 720:788 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 6 61 contain containing 737:746 arg2 acids 764:768 terminal sialic acids 748:768 terminal sialic acids 748:768 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 4 62 theme higher-energy 484:496 arg1 fragmentation 521:533 higher-energy collision dissociation fragmentation 484:533 higher-energy collision dissociation fragmentation 484:533 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 16 63 theme immune- 1631:1637 arg1 receptors 1652:1660 immune- or clearance receptors 1631:1660 immune- or clearance receptors containing carbohydrate recognition domains 1631:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 4 64 theme dissociation 508:519 arg1 fragmentation 521:533 higher-energy collision dissociation fragmentation 484:533 higher-energy collision dissociation fragmentation 484:533 Glycopeptides were enriched using zwitterionic HILIC zip-tips and analyzed by tandem mass spectrometry employing higher-energy collision dissociation fragmentation. 27574189 16 65 theme clearance 1642:1650 arg1 receptors 1652:1660 immune- or clearance receptors 1631:1660 immune- or clearance receptors containing carbohydrate recognition domains 1631:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 6 66 theme complex 724:730 arg1 type 732:735 the complex type 720:735 the complex type containing terminal sialic acids and fucose residues 720:788 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 5 67 theme electron 593:600 arg1 transfer 602:609 electron transfer 593:609 electron transfer dissociation fragmentation 593:636 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 1 68 theme glycoprotein 173:184 arg1 ADAMTS13 186:193 the plasma glycoprotein ADAMTS13 162:193 the plasma glycoprotein ADAMTS13 162:193 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 8 69 theme sialic 912:917 arg1 acid 919:922 sialic acid 912:922 sialic acid 912:922 Six O-linked glycans mostly terminating in sialic acid were found dispersed over ADAMTS13. 27574189 9 70 link O-linked 965:972 arg1 glycans 974:980 Five O-linked glycans 960:980 Five O-linked glycans 960:980 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 16 71 theme carbohydrate 1673:1684 arg1 domains 1698:1704 carbohydrate recognition domains 1673:1704 carbohydrate recognition domains 1673:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 5 72 theme dissociation 611:622 arg1 fragmentation 624:636 electron transfer dissociation fragmentation 593:636 electron transfer dissociation fragmentation 593:636 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 11 73 theme common 1098:1103 arg1 modification 1123:1134 a common posttranslational modification 1096:1134 a common posttranslational modification of thrombospondin type 1 repeats 1096:1167 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 11 73 theme common 1098:1103 arg1 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation 1078:1091 O-fucosylation is a common posttranslational modification of thrombospondin type 1 repeats. 27574189 6 74 theme glycans 704:710 arg1 majority 683:690 The majority 679:690 The majority of N-linked glycans 679:710 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 1 75 theme acquired 78:85 arg1 purpura 115:121 acquired thrombotic thrombocytopenic purpura 78:121 acquired thrombotic thrombocytopenic purpura 78:121 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 3 76 with peptides 325:332 arg1 chymotrypsin 357:368 chymotrypsin 357:368 chymotrypsin 357:368 Purified ADAMTS13 was reduced, alkylated, and processed into peptides with either trypsin or chymotrypsin. 27574189 3 76 with peptides 325:332 arg1 trypsin 346:352 trypsin 346:352 trypsin 346:352 Purified ADAMTS13 was reduced, alkylated, and processed into peptides with either trypsin or chymotrypsin. 27574189 0 77 from ADAMTS13 44:51 arg1 Identification 0:13 Identification 0:13 Identification of glycans on plasma-derived ADAMTS13. 0:52 Identification of glycans on plasma-derived ADAMTS13. 27574189 12 78 gly O-fucosylation 1186:1199 arg2 sites 1201:1205 7 O-fucosylation sites 1184:1205 7 O-fucosylation sites 1184:1205 We identified 7 O-fucosylation sites in the thrombospondin (TSP) type 1 repeats. 27574189 5 79 theme same 659:662 arg1 ion 674:676 the same precursor ion 655:676 the same precursor ion 655:676 Upon detection of a diagnostic ion of a glycan fragment, electron transfer dissociation fragmentation was performed on the same precursor ion. 27574189 1 80 theme thrombocytopenic 98:113 arg1 purpura 115:121 acquired thrombotic thrombocytopenic purpura 78:121 acquired thrombotic thrombocytopenic purpura 78:121 Patients suffering from acquired thrombotic thrombocytopenic purpura develop autoantibodies directed toward the plasma glycoprotein ADAMTS13. 27574189 16 81 with interaction 1614:1624 arg1 receptors 1652:1660 immune- or clearance receptors 1631:1660 immune- or clearance receptors containing carbohydrate recognition domains 1631:1704 Overall, our findings highlight the complexity of glycan modifications on ADAMTS13, which may have implications for its interaction with immune- or clearance receptors containing carbohydrate recognition domains. 27574189 8 82 link O-linked 873:880 arg1 glycans 882:888 Six O-linked glycans 869:888 Six O-linked glycans mostly terminating in sialic acid 869:922 Six O-linked glycans mostly terminating in sialic acid were found dispersed over ADAMTS13. 27574189 6 83 theme sialic 757:762 arg1 acids 764:768 terminal sialic acids 748:768 terminal sialic acids 748:768 The majority of N-linked glycans were of the complex type containing terminal sialic acids and fucose residues. 27574189 9 84 theme O-linked 965:972 arg1 glycans 974:980 Five O-linked glycans 960:980 Five O-linked glycans 960:980 Five O-linked glycans were attached to a Ser and one to Thr. 27574189 10 85 theme O-linked 1027:1034 arg1 glycans 1036:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans 1021:1042 All 6 O-linked glycans contained a terminal sialic acid. 27574189 14 86 theme proposed 1379:1386 arg1 sequence 1398:1405 the proposed consensus sequence 1375:1405 the proposed consensus sequence CSX(S/T)CG 1375:1416 This O-fucosylation site did not meet the proposed consensus sequence CSX(S/T)CG. 27574189 2 87 theme plasma-derived 239:252 arg1 ADAMTS13 254:261 plasma-derived ADAMTS13 239:261 plasma-derived ADAMTS13 239:261 Here, we studied the glycan composition of plasma-derived ADAMTS13. 27574189 8 88 dep found 929:933 arg1 dispersed 935:943 dispersed 935:943 dispersed over ADAMTS13 935:957 Six O-linked glycans mostly terminating in sialic acid were found dispersed over ADAMTS13. 27574189 7 89 theme spacer 854:859 arg1 domain 861:866 the spacer domain 850:866 the spacer domain 850:866 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 27574189 10 90 theme terminal 1056:1063 arg1 acid 1072:1075 a terminal sialic acid 1054:1075 a terminal sialic acid 1054:1075 All 6 O-linked glycans contained a terminal sialic acid. 27574189 7 91 attach attached 828:835 arg3 domain 861:866 the spacer domain 850:866 the spacer domain 850:866 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 27574189 7 91 attach attached 828:835 arg1 Asn614 840:845 Asn614 840:845 Asn614 840:845 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 27574189 7 91 attach attached 828:835 arg2 glycan 817:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan 791:822 A high mannose-containing glycan was attached to Asn614 in the spacer domain. 24327294 10 0 theme therapeutic 1681:1691 arg1 candidates 1693:1702 therapeutic candidates 1681:1702 therapeutic candidates 1681:1702 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 8 1 theme significant 1330:1340 arg1 decrease 1342:1349 a significant decrease 1328:1349 a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans 1328:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 3 2 theme polyethylene 509:520 arg1 glycol 522:527 polyethylene glycol 509:527 polyethylene glycol 509:527 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 2 3 theme hydrolase 289:297 arg1 administration 299:312 an exogenous cocaine hydrolase administration 268:312 an exogenous cocaine hydrolase administration 268:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 5 4 theme human 857:861 arg1 BChE 863:866 glycosylated human BChE 844:866 glycosylated human BChE 844:866 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 6 5 theme structures 1057:1066 arg1 flexibility 1031:1041 the flexibility 1027:1041 the flexibility of some local structures 1027:1066 Glycans did not change the overall stability of the BChE structure, but could increase the flexibility of some local structures. 24327294 9 6 theme better 1531:1536 arg1 chance 1538:1543 a better chance 1529:1543 a better chance to react with lysine residues 1529:1573 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 8 7 theme surrounding 1408:1418 arg1 glycans 1420:1426 their surrounding glycans 1402:1426 their surrounding glycans 1402:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 1 8 theme great 115:119 arg1 potential 121:129 great potential 115:129 great potential 115:129 Human butyrylcholinesterase (BChE) and its mutants have shown great potential in treating cocaine overdose and addiction. 24327294 2 9 theme long 242:245 arg1 period 247:252 a long period 240:252 a long period of time after an exogenous cocaine hydrolase administration 240:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 9 10 theme reaction 1471:1478 arg1 agents 1480:1485 PEGylation reaction agents 1460:1485 PEGylation reaction agents with smaller functional groups 1460:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 5 11 theme glycans 900:906 arg1 influence 887:895 the influence 883:895 the influence of glycans on the PEGylation modification 883:937 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 1 12 theme cocaine 143:149 arg1 overdose 151:158 cocaine overdose 143:158 cocaine overdose 143:158 Human butyrylcholinesterase (BChE) and its mutants have shown great potential in treating cocaine overdose and addiction. 24327294 9 13 theme functional 1500:1509 arg1 groups 1511:1516 smaller functional groups 1492:1516 smaller functional groups 1492:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 7 14 from sites 1137:1141 arg1 surface 1189:1195 the protein surface 1177:1195 the protein surface 1177:1195 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 9 15 with agents 1480:1485 arg1 groups 1511:1516 smaller functional groups 1492:1516 smaller functional groups 1492:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 8 16 theme lysine 1307:1312 arg1 residues 1314:1321 some lysine residues 1302:1321 some lysine residues 1302:1321 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 7 17 theme protein 1181:1187 arg1 surface 1189:1195 the protein surface 1177:1195 the protein surface 1177:1195 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 10 18 theme rational 1627:1634 arg1 engineering 1636:1646 rational engineering 1627:1646 rational engineering of human BChE and its mutants as therapeutic candidates 1627:1702 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 0 19 theme butyrylcholinesterase 30:50 arg1 model 2:6 A model 0:6 A model of glycosylated human butyrylcholinesterase 0:50 A model of glycosylated human butyrylcholinesterase. 24327294 6 20 theme structure 997:1005 arg1 stability 975:983 the overall stability 963:983 the overall stability of the BChE structure 963:1005 Glycans did not change the overall stability of the BChE structure, but could increase the flexibility of some local structures. 24327294 2 21 theme long 428:431 arg1 time 445:448 a sufficiently long circulation time 413:448 a sufficiently long circulation time 413:448 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 4 22 theme BChE 733:736 arg1 surface 709:715 the surface 705:715 the surface of glycosylated BChE 705:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 2 23 theme high 361:364 arg1 efficiency 376:385 a high catalytic efficiency 359:385 a high catalytic efficiency against cocaine 359:401 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 5 24 from influence 887:895 arg1 modification 926:937 the PEGylation modification 911:937 the PEGylation modification 911:937 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 5 25 theme PEGylation 915:924 arg1 modification 926:937 the PEGylation modification 911:937 the PEGylation modification 911:937 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 10 26 theme BChE 1657:1660 arg1 engineering 1636:1646 rational engineering 1627:1646 rational engineering of human BChE and its mutants as therapeutic candidates 1627:1702 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 7 27 theme sites 1137:1141 arg1 accessibility 1096:1108 the accessibility 1092:1108 the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface 1092:1195 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 0 28 theme human 24:28 arg1 butyrylcholinesterase 30:50 glycosylated human butyrylcholinesterase 11:50 glycosylated human butyrylcholinesterase 11:50 A model of glycosylated human butyrylcholinesterase. 24327294 3 29 mod modification 530:541 arg1 protein 560:566 a therapeutic protein 546:566 a therapeutic protein 546:566 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 3 29 mod modification 530:541 arg3 PEGylation 474:483 PEGylation 474:483 PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein 474:566 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 7 30 theme PEGylation 1117:1126 arg1 sites 1137:1141 the PEGylation reaction sites 1113:1141 the PEGylation reaction sites 1113:1141 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 30 theme PEGylation 1117:1126 arg1 residues 1164:1171 lysine residues 1157:1171 particularly lysine residues 1144:1171 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 0 31 gly glycosylated 11:22 arg1 butyrylcholinesterase 30:50 glycosylated human butyrylcholinesterase 11:50 glycosylated human butyrylcholinesterase 11:50 A model of glycosylated human butyrylcholinesterase. 24327294 9 32 theme lysine 1559:1564 arg1 residues 1566:1573 lysine residues 1559:1573 lysine residues 1559:1573 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 5 33 dep three-dimensional 812:828 arg1 3D 831:832 3D 831:832 3D 831:832 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 3 34 theme covalent 486:493 arg1 attachment 495:504 covalent attachment 486:504 covalent attachment of polyethylene glycol 486:527 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 1 35 theme Human 53:57 arg1 BChE 82:85 BChE 82:85 BChE 82:85 Human butyrylcholinesterase (BChE) and its mutants have shown great potential in treating cocaine overdose and addiction. 24327294 1 35 theme Human 53:57 arg1 butyrylcholinesterase 59:79 Human butyrylcholinesterase 53:79 Human butyrylcholinesterase (BChE) 53:86 Human butyrylcholinesterase (BChE) and its mutants have shown great potential in treating cocaine overdose and addiction. 24327294 7 36 theme Surface 1235:1241 arg1 SASAs 1250:1254 SASAs 1250:1254 SASAs 1250:1254 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 36 theme Surface 1235:1241 arg1 Areas 1243:1247 the Solvent Accessible Surface Areas 1212:1247 the Solvent Accessible Surface Areas (SASAs) of these residues 1212:1273 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 2 37 dep suppress 199:206 arg1 to 184:185 to 184:185 to 184:185 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 3 38 theme PEGylation 474:483 arg1 modification 530:541 PEGylation (covalent attachment of polyethylene glycol) modification 474:541 PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein 474:566 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 8 39 from decrease 1342:1349 arg1 SASA 1354:1357 SASA 1354:1357 SASA due to the direct or indirect influence of their surrounding glycans 1354:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 7 40 theme Solvent 1216:1222 arg1 SASAs 1250:1254 SASAs 1250:1254 SASAs 1250:1254 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 40 theme Solvent 1216:1222 arg1 Areas 1243:1247 the Solvent Accessible Surface Areas 1212:1247 the Solvent Accessible Surface Areas (SASAs) of these residues 1212:1273 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 3 41 theme glycol 522:527 arg1 attachment 495:504 covalent attachment 486:504 covalent attachment of polyethylene glycol 486:527 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 5 42 gly glycosylated 844:855 arg1 BChE 863:866 glycosylated human BChE 844:866 glycosylated human BChE 844:866 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 2 43 theme time 257:260 arg1 period 247:252 a long period 240:252 a long period of time after an exogenous cocaine hydrolase administration 240:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 5 44 theme glycosylated 844:855 arg1 BChE 863:866 glycosylated human BChE 844:866 glycosylated human BChE 844:866 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 4 45 link asparagine-linked 676:692 arg1 glycans 694:700 the asparagine-linked glycans 672:700 the asparagine-linked glycans on the surface of glycosylated BChE 672:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 2 46 theme cocaine 281:287 arg1 administration 299:312 an exogenous cocaine hydrolase administration 268:312 an exogenous cocaine hydrolase administration 268:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 5 47 theme three-dimensional 812:828 arg1 model 835:839 a three-dimensional (3D) model 810:839 a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification 810:937 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 3 48 theme therapeutic 548:558 arg1 protein 560:566 a therapeutic protein 546:566 a therapeutic protein 546:566 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 10 49 theme mutants 1670:1676 arg1 engineering 1636:1646 rational engineering 1627:1646 rational engineering of human BChE and its mutants as therapeutic candidates 1627:1702 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 8 50 theme glycans 1420:1426 arg1 influence 1389:1397 the direct or indirect influence 1366:1397 the direct or indirect influence of their surrounding glycans 1366:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 3 51 theme protein 560:566 arg1 modification 530:541 PEGylation (covalent attachment of polyethylene glycol) modification 474:541 PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein 474:566 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 7 52 from surface 1189:1195 arg1 accessibility 1096:1108 the accessibility 1092:1108 the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface 1092:1195 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 9 53 theme PEGylation 1460:1469 arg1 agents 1480:1485 PEGylation reaction agents 1460:1485 PEGylation reaction agents with smaller functional groups 1460:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 3 54 theme biological 584:593 arg1 half-life 595:603 the biological half-life 580:603 the biological half-life of the protein 580:618 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 5 55 theme BChE 863:866 arg1 model 835:839 a three-dimensional (3D) model 810:839 a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification 810:937 In this study, we built a three-dimensional (3D) model of glycosylated human BChE to investigate the influence of glycans on the PEGylation modification. 24327294 6 56 theme BChE 992:995 arg1 structure 997:1005 the BChE structure 988:1005 the BChE structure 988:1005 Glycans did not change the overall stability of the BChE structure, but could increase the flexibility of some local structures. 24327294 9 57 theme smaller 1492:1498 arg1 groups 1511:1516 smaller functional groups 1492:1516 smaller functional groups 1492:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 4 58 theme asparagine-linked 676:692 arg1 glycans 694:700 the asparagine-linked glycans 672:700 the asparagine-linked glycans on the surface of glycosylated BChE 672:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 4 59 gly glycosylated 720:731 arg1 BChE 733:736 glycosylated BChE 720:736 glycosylated BChE 720:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 2 60 theme exogenous 271:279 arg1 administration 299:312 an exogenous cocaine hydrolase administration 268:312 an exogenous cocaine hydrolase administration 268:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 6 61 theme overall 967:973 arg1 stability 975:983 the overall stability 963:983 the overall stability of the BChE structure 963:1005 Glycans did not change the overall stability of the BChE structure, but could increase the flexibility of some local structures. 24327294 2 62 contain have 345:348 arg2 time 445:448 a sufficiently long circulation time 413:448 a sufficiently long circulation time 413:448 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 2 62 contain have 345:348 arg1 enzyme 331:336 the therapeutic enzyme 315:336 the therapeutic enzyme 315:336 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 2 62 contain have 345:348 arg2 efficiency 376:385 a high catalytic efficiency 359:385 a high catalytic efficiency against cocaine 359:401 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 2 63 from reward 216:221 arg1 brain 230:234 the brain 226:234 the brain 226:234 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 6 64 theme local 1051:1055 arg1 structures 1057:1066 some local structures 1046:1066 some local structures 1046:1066 Glycans did not change the overall stability of the BChE structure, but could increase the flexibility of some local structures. 24327294 4 65 theme PEGylation 761:770 arg1 modification 772:783 the PEGylation modification 757:783 the PEGylation modification 757:783 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 2 66 theme catalytic 366:374 arg1 efficiency 376:385 a high catalytic efficiency 359:385 a high catalytic efficiency against cocaine 359:401 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 7 67 theme lysine 1157:1162 arg1 sites 1137:1141 the PEGylation reaction sites 1113:1141 the PEGylation reaction sites 1113:1141 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 67 theme lysine 1157:1162 arg1 residues 1164:1171 lysine residues 1157:1171 particularly lysine residues 1144:1171 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 3 68 dep PEGylation 474:483 arg1 attachment 495:504 covalent attachment 486:504 covalent attachment of polyethylene glycol 486:527 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 4 69 theme glycosylated 720:731 arg1 BChE 733:736 glycosylated BChE 720:736 glycosylated BChE 720:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 24327294 10 70 theme structural 1606:1615 arg1 basis 1617:1621 a structural basis 1604:1621 a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates 1604:1702 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 0 71 theme glycosylated 11:22 arg1 butyrylcholinesterase 30:50 glycosylated human butyrylcholinesterase 11:50 glycosylated human butyrylcholinesterase 11:50 A model of glycosylated human butyrylcholinesterase. 24327294 7 72 theme residues 1266:1273 arg1 SASAs 1250:1254 SASAs 1250:1254 SASAs 1250:1254 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 72 theme residues 1266:1273 arg1 Areas 1243:1247 the Solvent Accessible Surface Areas 1212:1247 the Solvent Accessible Surface Areas (SASAs) of these residues 1212:1273 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 10 73 theme human 1651:1655 arg1 BChE 1657:1660 human BChE 1651:1660 human BChE 1651:1660 This investigation provides a structural basis for rational engineering of human BChE and its mutants as therapeutic candidates. 24327294 7 74 theme reaction 1128:1135 arg1 sites 1137:1141 the PEGylation reaction sites 1113:1141 the PEGylation reaction sites 1113:1141 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 74 theme reaction 1128:1135 arg1 residues 1164:1171 lysine residues 1157:1171 particularly lysine residues 1144:1171 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 3 75 theme protein 612:618 arg1 half-life 595:603 the biological half-life 580:603 the biological half-life of the protein 580:618 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 2 76 theme circulation 433:443 arg1 time 445:448 a sufficiently long circulation time 413:448 a sufficiently long circulation time 413:448 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 7 77 from accessibility 1096:1108 arg1 surface 1189:1195 the protein surface 1177:1195 the protein surface 1177:1195 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 2 78 theme cocaine 208:214 arg1 reward 216:221 cocaine reward 208:221 cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration 208:312 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 2 79 theme therapeutic 319:329 arg1 enzyme 331:336 the therapeutic enzyme 315:336 the therapeutic enzyme 315:336 In order to effectively suppress cocaine reward in the brain for a long period of time after an exogenous cocaine hydrolase administration, the therapeutic enzyme should have not only a high catalytic efficiency against cocaine, but also a sufficiently long circulation time. 24327294 3 80 theme biological 642:651 arg1 function 653:660 its biological function 638:660 its biological function 638:660 It has been known that PEGylation (covalent attachment of polyethylene glycol) modification of a therapeutic protein can prolong the biological half-life of the protein without affecting its biological function. 24327294 8 81 theme direct 1370:1375 arg1 influence 1389:1397 the direct or indirect influence 1366:1397 the direct or indirect influence of their surrounding glycans 1366:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 7 82 theme Accessible 1224:1233 arg1 SASAs 1250:1254 SASAs 1250:1254 SASAs 1250:1254 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 7 82 theme Accessible 1224:1233 arg1 Areas 1243:1247 the Solvent Accessible Surface Areas 1212:1247 the Solvent Accessible Surface Areas (SASAs) of these residues 1212:1273 For further evaluating the accessibility of the PEGylation reaction sites, particularly lysine residues, on the protein surface, we calculated the Solvent Accessible Surface Areas (SASAs) of these residues. 24327294 9 83 contain have 1524:1527 arg2 chance 1538:1543 a better chance 1529:1543 a better chance to react with lysine residues 1529:1573 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 9 83 contain have 1524:1527 arg1 agents 1480:1485 PEGylation reaction agents 1460:1485 PEGylation reaction agents with smaller functional groups 1460:1516 The results also indicate that PEGylation reaction agents with smaller functional groups could have a better chance to react with lysine residues. 24327294 8 84 theme indirect 1380:1387 arg1 influence 1389:1397 the direct or indirect influence 1366:1397 the direct or indirect influence of their surrounding glycans 1366:1426 The results indicate that some lysine residues show a significant decrease in SASA due to the direct or indirect influence of their surrounding glycans. 24327294 4 85 from glycans 694:700 arg1 surface 709:715 the surface 705:715 the surface of glycosylated BChE 705:736 However, the asparagine-linked glycans on the surface of glycosylated BChE may interfere with the PEGylation modification. 26192331 2 0 theme oligo-polysialic 672:687 arg1 products 700:707 oligo-polysialic acid chain products 672:707 oligo-polysialic acid chain products 672:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 1 theme extended 617:624 arg1 groove 650:655 an extended electropositive surface groove 614:655 an extended electropositive surface groove 614:655 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 2 theme surface-localized 188:204 arg1 glycoproteins 206:218 surface-localized glycoproteins 188:218 surface-localized glycoproteins 188:218 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 3 theme nonreducing 297:307 arg1 ends 309:312 the nonreducing ends 293:312 the nonreducing ends of sialic acid acceptors 293:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 4 theme sugar 756:760 arg1 analog 762:767 sugar analog 756:767 sugar analog 756:767 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 1 5 theme glycoproteins 206:218 arg1 oligo- 158:163 oligo- 158:163 oligo- 158:163 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 5 theme glycoproteins 206:218 arg1 polysialylation 169:183 polysialylation 169:183 polysialylation 169:183 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 2 6 theme motifs 581:586 arg1 cluster 528:534 a cluster 526:534 a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products 526:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 7 theme ST8Sia 135:140 arg1 family 142:147 the mammalian ST8Sia family 121:147 the mammalian ST8Sia family 121:147 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 8 theme substrate 896:904 arg1 binding 906:912 substrate binding 896:912 substrate binding 896:912 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 9 theme structural 570:579 arg1 motifs 581:586 polysialyltransferase-specific structural motifs 539:586 polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products 539:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 10 from species 502:508 arg1 polysialyltransferase 471:491 a polysialyltransferase 469:491 a polysialyltransferase from any species 469:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 10 from species 502:508 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 10 from species 502:508 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 11 from resolution 391:400 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 11 from resolution 391:400 arg1 ST8SiaIII 371:379 human ST8SiaIII 365:379 human ST8SiaIII at 1.85-Å resolution 365:400 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 11 from resolution 391:400 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 12 theme sialic 317:322 arg1 acceptors 329:337 sialic acid acceptors 317:337 sialic acid acceptors 317:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 2 13 theme polysialyltransferase-specific 539:568 arg1 motifs 581:586 polysialyltransferase-specific structural motifs 539:586 polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products 539:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 14 theme solved 449:454 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 14 theme solved 449:454 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 3 15 dep donor 750:754 arg1 analog 762:767 sugar analog 756:767 sugar analog 756:767 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 3 16 theme sulfated 775:782 arg1 acceptor 791:798 a sulfated glycan acceptor 773:798 a sulfated glycan acceptor identified with a sialyltransferase glycan array 773:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 17 theme products 700:707 arg1 binding 661:667 binding 661:667 binding of oligo-polysialic acid chain products 661:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 18 theme glycolipids 224:234 arg1 oligo- 158:163 oligo- 158:163 oligo- 158:163 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 18 theme glycolipids 224:234 arg1 polysialylation 169:183 polysialylation 169:183 polysialylation 169:183 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 19 theme glycan 784:789 arg1 acceptor 791:798 a sulfated glycan acceptor 773:798 a sulfated glycan acceptor identified with a sialyltransferase glycan array 773:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 20 theme chain 694:698 arg1 products 700:707 oligo-polysialic acid chain products 672:707 oligo-polysialic acid chain products 672:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 3 21 theme sialyltransferase 818:834 arg1 array 843:847 a sialyltransferase glycan array 816:847 a sialyltransferase glycan array 816:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 22 theme human 365:369 arg1 ST8SiaIII 371:379 human ST8SiaIII 365:379 human ST8SiaIII at 1.85-Å resolution 365:400 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 23 theme surface 642:648 arg1 groove 650:655 an extended electropositive surface groove 614:655 an extended electropositive surface groove 614:655 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 24 theme polysialyltransferase 471:491 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 24 theme polysialyltransferase 471:491 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 25 theme acid 324:327 arg1 acceptors 329:337 sialic acid acceptors 317:337 sialic acid acceptors 317:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 2 26 theme acid 689:692 arg1 products 700:707 oligo-polysialic acid chain products 672:707 oligo-polysialic acid chain products 672:707 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 0 27 theme ST8SiaIII 19:27 arg1 sialyltransferase 29:45 human ST8SiaIII sialyltransferase 13:45 human ST8SiaIII sialyltransferase 13:45 Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation. 26192331 1 28 theme acceptors 329:337 arg1 ends 309:312 the nonreducing ends 293:312 the nonreducing ends of sialic acid acceptors 293:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 29 theme mammalian 125:133 arg1 family 142:147 the mammalian ST8Sia family 121:147 the mammalian ST8Sia family 121:147 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 0 30 theme human 13:17 arg1 sialyltransferase 29:45 human ST8SiaIII sialyltransferase 13:45 human ST8SiaIII sialyltransferase 13:45 Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation. 26192331 2 31 theme crystal 344:350 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 31 theme crystal 344:350 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 3 32 with complex 722:728 arg1 acceptor 791:798 a sulfated glycan acceptor 773:798 a sulfated glycan acceptor identified with a sialyltransferase glycan array 773:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 3 32 with complex 722:728 arg1 donor 750:754 a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array 748:847 donor 750:754 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 1 33 theme sialic 256:261 arg1 acids 263:267 sialic acids 256:267 sialic acids from CMP-sialic acid 256:288 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 2 34 theme first 443:447 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 34 theme first 443:447 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 35 theme acids 263:267 arg1 transfer 244:251 transfer 244:251 transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors 244:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 0 36 theme sialyltransferase 29:45 arg1 Structure 0:8 Structure 0:8 Structure of human ST8SiaIII sialyltransferase 0:45 Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation. 26192331 3 37 theme glycan 836:841 arg1 array 843:847 a sialyltransferase glycan array 816:847 a sialyltransferase glycan array 816:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 38 theme electropositive 626:640 arg1 groove 650:655 an extended electropositive surface groove 614:655 an extended electropositive surface groove 614:655 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 39 theme family 142:147 arg1 Sialyltransferases 99:116 Sialyltransferases 99:116 Sialyltransferases of the mammalian ST8Sia family 99:147 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 40 gly polysialylation 169:183 arg1 glycolipids 224:234 glycolipids 224:234 glycolipids 224:234 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 40 gly polysialylation 169:183 arg1 glycoproteins 206:218 surface-localized glycoproteins 188:218 surface-localized glycoproteins 188:218 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 41 theme ternary 714:720 arg1 complex 722:728 The ternary complex 710:728 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array 710:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 42 theme 1.85-Å 384:389 arg1 resolution 391:400 1.85-Å resolution 384:400 1.85-Å resolution 384:400 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 43 gly glycoproteins 206:218 arg1 glycoproteins 206:218 surface-localized glycoproteins 188:218 surface-localized glycoproteins 188:218 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 44 theme CMP-sialic 274:283 arg1 acid 285:288 CMP-sialic acid 274:288 CMP-sialic acid 274:288 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 45 from acid 285:288 arg1 transfer 244:251 transfer 244:251 transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors 244:337 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 1 45 from acid 285:288 arg1 acids 263:267 sialic acids 256:267 sialic acids from CMP-sialic acid 256:288 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 46 theme ST8SiaIII 733:741 arg1 complex 722:728 The ternary complex 710:728 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array 710:847 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 2 47 theme ST8SiaIII 371:379 arg1 structure 352:360 The crystal structure 340:360 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here 340:415 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 47 theme ST8SiaIII 371:379 arg1 structure 456:464 the first solved structure 439:464 the first solved structure of a polysialyltransferase from any species 439:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 1 48 from transfer 244:251 arg1 acid 285:288 CMP-sialic acid 274:288 CMP-sialic acid 274:288 Sialyltransferases of the mammalian ST8Sia family catalyze oligo- and polysialylation of surface-localized glycoproteins and glycolipids through transfer of sialic acids from CMP-sialic acid to the nonreducing ends of sialic acid acceptors. 26192331 3 49 theme sialyl 931:936 arg1 transfer 938:945 sialyl transfer 931:945 sialyl transfer 931:945 The ternary complex of ST8SiaIII with a donor sugar analog and a sulfated glycan acceptor identified with a sialyltransferase glycan array provides insight into the residues involved in substrate binding, specificity and sialyl transfer. 26192331 0 50 theme cell-surface 69:80 arg1 polysialylation 82:96 cell-surface polysialylation 69:96 cell-surface polysialylation 69:96 Structure of human ST8SiaIII sialyltransferase provides insight into cell-surface polysialylation. 26192331 2 51 from structure 456:464 arg1 species 502:508 any species 498:508 any species 498:508 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 26192331 2 52 from structure 352:360 arg1 resolution 391:400 1.85-Å resolution 384:400 1.85-Å resolution 384:400 The crystal structure of human ST8SiaIII at 1.85-Å resolution presented here is, to our knowledge, the first solved structure of a polysialyltransferase from any species, and it reveals a cluster of polysialyltransferase-specific structural motifs that collectively provide an extended electropositive surface groove for binding of oligo-polysialic acid chain products. 27818199 0 0 theme angiotensin-converting 68:89 arg1 enzyme 91:96 human angiotensin-converting enzyme 62:96 human angiotensin-converting enzyme 62:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 13 1 theme cleavage 1974:1981 arg1 site 1983:1986 the cleavage site 1970:1986 the cleavage site 1970:1986 Thus, the conformational flexibility that the leucine confers to the stalk, is increased by the lack of glycosylation reducing access of the sheddase to the cleavage site. 27818199 4 2 theme secondary 608:616 arg1 α-helix 630:636 α-helix 7 and 8 630:644 α-helix 7 and 8 630:644 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 4 2 theme secondary 608:616 arg1 structures 618:627 discrete secondary structures 599:627 discrete secondary structures (α-helix 7 and 8) on the surface of tACE 599:668 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 11 3 theme tACE 1599:1602 arg1 mutant 1604:1609 The P623L tACE mutant 1589:1609 The P623L tACE mutant 1589:1609 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 2 4 theme identical 299:307 arg1 domain 346:351 transmembrane and cytoplasmic domain 316:351 transmembrane and cytoplasmic domain 316:351 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 2 4 theme identical 299:307 arg1 stalk 309:313 an identical stalk 296:313 an identical stalk 296:313 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 1 5 theme smaller 214:220 arg1 tACE 240:243 tACE 240:243 tACE 240:243 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 5 theme smaller 214:220 arg1 identical 249:257 identical 249:257 identical 249:257 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 5 theme smaller 214:220 arg1 isoform 231:237 the smaller germinal isoform 210:237 the smaller germinal isoform (tACE) 210:244 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 0 6 theme human 62:66 arg1 enzyme 91:96 human angiotensin-converting enzyme 62:96 human angiotensin-converting enzyme 62:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 0 7 from effect 4:9 arg1 shedding 50:57 the ectodomain shedding 35:57 the ectodomain shedding of human angiotensin-converting enzyme 35:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 12 8 theme N-linked 1742:1749 arg1 glycan 1751:1756 an N-linked glycan 1739:1756 an N-linked glycan 1739:1756 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 4 9 theme discrete 599:606 arg1 α-helix 630:636 α-helix 7 and 8 630:644 α-helix 7 and 8 630:644 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 4 9 theme discrete 599:606 arg1 structures 618:627 discrete secondary structures 599:627 discrete secondary structures (α-helix 7 and 8) on the surface of tACE 599:668 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 1 10 theme germinal 222:229 arg1 tACE 240:243 tACE 240:243 tACE 240:243 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 10 theme germinal 222:229 arg1 identical 249:257 identical 249:257 identical 249:257 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 10 theme germinal 222:229 arg1 isoform 231:237 the smaller germinal isoform 210:237 the smaller germinal isoform (tACE) 210:244 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 7 11 theme mutation 975:982 arg1 extension 957:965 An N-terminal extension 943:965 An N-terminal extension of this mutation 943:982 An N-terminal extension of this mutation caused a reduction in cellular ACE activity. 27818199 10 12 from mutation 1554:1561 arg1 tACE 1566:1569 tACE 1566:1569 tACE 1566:1569 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 13 13 theme sheddase 1958:1965 arg1 access 1944:1949 access 1944:1949 access of the sheddase to the cleavage site 1944:1986 Thus, the conformational flexibility that the leucine confers to the stalk, is increased by the lack of glycosylation reducing access of the sheddase to the cleavage site. 27818199 0 14 theme enzyme 91:96 arg1 shedding 50:57 the ectodomain shedding 35:57 the ectodomain shedding of human angiotensin-converting enzyme 35:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 13 15 theme conformational 1827:1840 arg1 flexibility 1842:1852 the conformational flexibility 1823:1852 the conformational flexibility 1823:1852 Thus, the conformational flexibility that the leucine confers to the stalk, is increased by the lack of glycosylation reducing access of the sheddase to the cleavage site. 27818199 10 16 theme asparagine 1454:1463 arg1 glycosylation 1433:1445 glycosylation 1433:1445 glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE 1433:1512 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 3 17 theme proximal 477:484 arg1 regions 458:464 regions 458:464 regions distal and proximal 458:484 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 5 18 theme absolute 780:787 arg1 requirement 789:799 an absolute requirement 777:799 an absolute requirement for shedding 777:812 Surprisingly, neither α-helix 7 nor α-helix 8 proved to be an absolute requirement for shedding. 27818199 5 18 theme absolute 780:787 arg1 α-helix 740:746 α-helix 7 740:748 α-helix 7 740:748 Surprisingly, neither α-helix 7 nor α-helix 8 proved to be an absolute requirement for shedding. 27818199 5 18 theme absolute 780:787 arg1 α-helix 754:760 α-helix 8 754:762 α-helix 8 754:762 Surprisingly, neither α-helix 7 nor α-helix 8 proved to be an absolute requirement for shedding. 27818199 12 19 theme glycan 1751:1756 arg1 absence 1728:1734 The absence 1724:1734 The absence 1724:1734 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 3 20 theme regions 458:464 arg1 role 450:453 the role 446:453 the role of regions distal and proximal to the cleavage site in human ACE shedding 446:527 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 12 21 from increase 1795:1802 arg1 shedding 1807:1814 shedding 1807:1814 shedding 1807:1814 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 8 22 theme protein 1081:1087 arg1 processing 1063:1072 the processing 1059:1072 the processing of the protein to the membrane 1059:1103 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 11 23 theme P623L 1593:1597 arg1 mutant 1604:1609 The P623L tACE mutant 1589:1609 The P623L tACE mutant 1589:1609 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 8 24 theme form 1154:1157 arg1 expression 1119:1128 expression 1119:1128 expression of an underglycosylated form of ACE 1119:1164 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 3 25 theme distal 466:471 arg1 regions 458:464 regions 458:464 regions distal and proximal 458:484 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 2 26 theme cytoplasmic 334:344 arg1 domain 346:351 transmembrane and cytoplasmic domain 316:351 transmembrane and cytoplasmic domain 316:351 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 2 26 theme cytoplasmic 334:344 arg1 stalk 309:313 an identical stalk 296:313 an identical stalk 296:313 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 12 27 theme greater 1787:1793 arg1 increase 1795:1802 an even greater increase 1779:1802 an even greater increase in shedding 1779:1814 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 7 28 theme N-terminal 946:955 arg1 extension 957:965 An N-terminal extension 943:965 An N-terminal extension of this mutation 943:982 An N-terminal extension of this mutation caused a reduction in cellular ACE activity. 27818199 1 29 dep comprised 147:155 arg1 whereas 202:208 whereas 202:208 whereas 202:208 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 12 30 link N-linked 1742:1749 arg1 glycan 1751:1756 an N-linked glycan 1739:1756 an N-linked glycan 1739:1756 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 10 31 theme Pro1199-Leu 1481:1491 arg1 polymorphism 1493:1504 the Pro1199-Leu polymorphism 1477:1504 the Pro1199-Leu polymorphism in sACE 1477:1512 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 6 32 theme residues 850:857 arg1 ectodomain 831:840 the proximal ectodomain 818:840 the proximal ectodomain of tACE residues 818:857 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 6 32 theme residues 850:857 arg1 residues 850:857 tACE residues 845:857 tACE residues 845:857 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 2 33 theme transmembrane 316:328 arg1 domain 346:351 transmembrane and cytoplasmic domain 316:351 transmembrane and cytoplasmic domain 316:351 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 2 33 theme transmembrane 316:328 arg1 stalk 309:313 an identical stalk 296:313 an identical stalk 296:313 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 0 34 theme structural 14:23 arg1 motifs 25:30 structural motifs 14:30 structural motifs 14:30 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 4 35 dep N 582:582 arg1 domains 590:596 domains 590:596 domains 590:596 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 4 35 dep N 582:582 arg1 the 578:580 the 578:580 the 578:580 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 6 36 from decrease 917:924 arg1 shedding 933:940 ACE shedding 929:940 ACE shedding 929:940 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 1 37 theme Somatic 99:105 arg1 sACE 138:141 sACE 138:141 sACE 138:141 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 37 theme Somatic 99:105 arg1 enzyme 130:135 Somatic angiotensin converting enzyme 99:135 Somatic angiotensin converting enzyme (sACE) 99:142 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 3 38 theme ACE 516:518 arg1 shedding 520:527 human ACE shedding 510:527 human ACE shedding 510:527 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 2 39 theme ectodomain 366:375 arg1 shedding 377:384 ectodomain shedding 366:384 ectodomain shedding by an as yet unknown protease 366:414 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 4 40 from structures 618:627 arg1 surface 654:660 the surface 650:660 the surface of tACE 650:668 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 4 41 theme tACE 665:668 arg1 surface 654:660 the surface 650:660 the surface of tACE 650:668 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 1 42 theme angiotensin 107:117 arg1 sACE 138:141 sACE 138:141 sACE 138:141 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 42 theme angiotensin 107:117 arg1 enzyme 130:135 Somatic angiotensin converting enzyme 99:135 Somatic angiotensin converting enzyme (sACE) 99:142 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 12 43 from N620 1761:1764 arg1 absence 1728:1734 The absence 1724:1734 The absence 1724:1734 The absence of an N-linked glycan at N620, resulted in an even greater increase in shedding. 27818199 7 44 theme ACE 1015:1017 arg1 activity 1019:1026 cellular ACE activity 1006:1026 cellular ACE activity 1006:1026 An N-terminal extension of this mutation caused a reduction in cellular ACE activity. 27818199 9 45 theme homologous 1201:1210 arg1 region 1212:1217 the homologous region 1197:1217 the homologous region of the N domain 1197:1233 When E608-H614 was mutated to the homologous region of the N domain, processing was normal and shedding only moderately decreased suggesting that this region is more crucial for the processing of ACE than it is for regulating shedding. 27818199 11 46 gly glycosylated 1710:1721 arg1 N620WT 1699:1704 N620WT 1699:1704 N620WT 1699:1704 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 1 47 dep domains 175:181 arg1 domains 175:181 two homologous domains 160:181 two homologous domains (N and C domains) 160:199 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 47 dep domains 175:181 arg1 C 190:190 C 190:190 C 190:190 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 47 dep domains 175:181 arg1 N 184:184 N 184:184 N 184:184 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 3 48 from site 502:505 arg1 shedding 520:527 human ACE shedding 510:527 human ACE shedding 510:527 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 0 49 theme motifs 25:30 arg1 effect 4:9 The effect 0:9 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme 0:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 6 50 theme proximal 822:829 arg1 ectodomain 831:840 the proximal ectodomain 818:840 the proximal ectodomain of tACE residues 818:857 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 6 50 theme proximal 822:829 arg1 residues 850:857 tACE residues 845:857 tACE residues 845:857 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 9 51 theme N 1226:1226 arg1 domain 1228:1233 the N domain 1222:1233 the N domain 1222:1233 When E608-H614 was mutated to the homologous region of the N domain, processing was normal and shedding only moderately decreased suggesting that this region is more crucial for the processing of ACE than it is for regulating shedding. 27818199 11 52 theme tryptic 1669:1675 arg1 digest 1677:1682 a tryptic digest 1667:1682 a tryptic digest 1667:1682 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 4 53 theme N 695:695 arg1 counterparts 704:715 their N domain counterparts 689:715 their N domain counterparts 689:715 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 4 54 theme domain 697:702 arg1 counterparts 704:715 their N domain counterparts 689:715 their N domain counterparts 689:715 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 2 55 theme unknown 399:405 arg1 protease 407:414 an as yet unknown protease 389:414 an as yet unknown protease 389:414 Both isozymes share an identical stalk, transmembrane and cytoplasmic domain, and undergo ectodomain shedding by an as yet unknown protease. 27818199 6 56 theme tACE 845:848 arg1 residues 850:857 tACE residues 845:857 tACE residues 845:857 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 7 57 from reduction 993:1001 arg1 activity 1019:1026 cellular ACE activity 1006:1026 cellular ACE activity 1006:1026 An N-terminal extension of this mutation caused a reduction in cellular ACE activity. 27818199 1 58 theme converting 119:128 arg1 sACE 138:141 sACE 138:141 sACE 138:141 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 58 theme converting 119:128 arg1 enzyme 130:135 Somatic angiotensin converting enzyme 99:135 Somatic angiotensin converting enzyme (sACE) 99:142 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 7 59 theme cellular 1006:1013 arg1 activity 1019:1026 cellular ACE activity 1006:1026 cellular ACE activity 1006:1026 An N-terminal extension of this mutation caused a reduction in cellular ACE activity. 27818199 1 60 theme C 266:266 arg1 domain 268:273 the C domain 262:273 the C domain 262:273 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 3 61 theme human 510:514 arg1 shedding 520:527 human ACE shedding 510:527 human ACE shedding 510:527 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 8 62 theme underglycosylated 1136:1152 arg1 form 1154:1157 an underglycosylated form 1133:1157 an underglycosylated form of ACE 1133:1164 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 3 63 theme cleavage 493:500 arg1 site 502:505 the cleavage site 489:505 the cleavage site in human ACE shedding 489:527 Here we present evidence for the role of regions distal and proximal to the cleavage site in human ACE shedding. 27818199 10 64 gly glycosylation 1433:1445 arg1 asparagine 1454:1463 the asparagine 1450:1463 the asparagine proximal to the Pro1199-Leu polymorphism in sACE 1450:1512 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 10 65 from polymorphism 1493:1504 arg1 sACE 1509:1512 sACE 1509:1512 sACE 1509:1512 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 8 66 theme ACE 1162:1164 arg1 form 1154:1157 an underglycosylated form 1133:1157 an underglycosylated form of ACE 1133:1164 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 9 67 theme domain 1228:1233 arg1 region 1212:1217 the homologous region 1197:1217 the homologous region of the N domain 1197:1233 When E608-H614 was mutated to the homologous region of the N domain, processing was normal and shedding only moderately decreased suggesting that this region is more crucial for the processing of ACE than it is for regulating shedding. 27818199 6 68 theme ACE 929:931 arg1 shedding 933:940 ACE shedding 929:940 ACE shedding 929:940 In the proximal ectodomain of tACE residues H610-L614 were mutated to alanines and this resulted in a decrease in ACE shedding. 27818199 11 69 theme MALDI 1646:1650 arg1 analysis 1655:1662 MALDI MS analysis 1646:1662 MALDI MS analysis of a tryptic digest 1646:1682 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 11 70 theme digest 1677:1682 arg1 analysis 1655:1662 MALDI MS analysis 1646:1662 MALDI MS analysis of a tryptic digest 1646:1682 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 11 71 theme MS 1652:1653 arg1 analysis 1655:1662 MALDI MS analysis 1646:1662 MALDI MS analysis of a tryptic digest 1646:1682 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 10 72 theme equivalent 1537:1546 arg1 mutation 1554:1561 the equivalent P623L mutation 1533:1561 the equivalent P623L mutation in tACE 1533:1569 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 11 73 from increase 1621:1628 arg1 shedding 1633:1640 shedding 1633:1640 shedding 1633:1640 The P623L tACE mutant showed an increase in shedding and MALDI MS analysis of a tryptic digest indicated that N620WT was glycosylated. 27818199 10 74 theme P623L 1548:1552 arg1 mutation 1554:1561 the equivalent P623L mutation 1533:1561 the equivalent P623L mutation in tACE 1533:1569 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 27818199 1 75 dep N 184:184 arg1 domains 192:198 domains 192:198 domains 192:198 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 4 76 theme intrinsic 548:556 arg1 differences 558:568 intrinsic differences 548:568 intrinsic differences between the N and C domains 548:596 First, because of intrinsic differences between the N and C domains, discrete secondary structures (α-helix 7 and 8) on the surface of tACE were replaced with their N domain counterparts. 27818199 1 77 theme homologous 164:173 arg1 domains 175:181 two homologous domains 160:181 two homologous domains (N and C domains) 160:199 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 77 theme homologous 164:173 arg1 C 190:190 C 190:190 C 190:190 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 1 77 theme homologous 164:173 arg1 N 184:184 N 184:184 N 184:184 Somatic angiotensin converting enzyme (sACE) is comprised of two homologous domains (N and C domains), whereas the smaller germinal isoform (tACE) is identical to the C domain. 27818199 0 78 theme ectodomain 39:48 arg1 shedding 50:57 the ectodomain shedding 35:57 the ectodomain shedding of human angiotensin-converting enzyme 35:96 The effect of structural motifs on the ectodomain shedding of human angiotensin-converting enzyme. 27818199 8 79 gly underglycosylated 1136:1152 arg1 ACE 1162:1164 ACE 1162:1164 ACE 1162:1164 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 8 79 gly underglycosylated 1136:1152 arg1 form 1154:1157 an underglycosylated form 1133:1157 an underglycosylated form of ACE 1133:1164 More importantly, it affected the processing of the protein to the membrane, resulting in expression of an underglycosylated form of ACE. 27818199 13 80 theme glycosylation 1921:1933 arg1 lack 1913:1916 the lack 1909:1916 the lack of glycosylation reducing access of the sheddase to the cleavage site 1909:1986 Thus, the conformational flexibility that the leucine confers to the stalk, is increased by the lack of glycosylation reducing access of the sheddase to the cleavage site. 27818199 9 81 theme ACE 1363:1365 arg1 processing 1349:1358 the processing 1345:1358 the processing of ACE 1345:1365 When E608-H614 was mutated to the homologous region of the N domain, processing was normal and shedding only moderately decreased suggesting that this region is more crucial for the processing of ACE than it is for regulating shedding. 27818199 10 82 theme proximal 1465:1472 arg1 asparagine 1454:1463 the asparagine 1450:1463 the asparagine proximal to the Pro1199-Leu polymorphism in sACE 1450:1512 Finally, to determine whether glycosylation of the asparagine proximal to the Pro1199-Leu polymorphism in sACE affected shedding, the equivalent P623L mutation in tACE was investigated. 26479949 10 0 theme gene 1851:1854 arg1 products 1856:1863 93 gene products 1848:1863 93 gene products 1848:1863 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 2 1 theme maltose 389:395 arg1 NPs 433:435 NPs 433:435 NPs 433:435 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 1 theme maltose 389:395 arg1 nanoparticles 418:430 hydrophilic maltose functionalized Fe3O4 nanoparticles 377:430 hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 377:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 10 2 theme glycosylation 1873:1885 arg1 sites 1887:1891 124 glycosylation sites 1869:1891 124 glycosylation sites 1869:1891 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 0 3 theme Click 0:4 arg1 Synthesis 6:14 Click Synthesis 0:14 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 0:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 6 4 theme angle 1154:1158 arg1 measurement 1160:1170 water contact angle measurement 1140:1170 water contact angle measurement 1140:1170 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 2 5 theme surface 336:342 arg1 strategy 357:364 an easy and one-step surface modification strategy 315:364 an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 315:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 10 6 gly glycopeptides 1820:1832 arg2 glycopeptides 1820:1832 115 different N-linked glycopeptides 1797:1832 115 different N-linked glycopeptides 1797:1832 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 6 7 theme water 1140:1144 arg1 measurement 1160:1170 water contact angle measurement 1140:1170 water contact angle measurement 1140:1170 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 4 8 theme Fe3O4 674:678 arg1 NPs 699:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 9 9 gly glycopeptides 1675:1687 arg2 glycopeptides 1675:1687 glycopeptides 1675:1687 glycopeptides from human renal mesangial cells (HRMC) 1675:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 9 9 gly glycopeptides 1675:1687 arg1 HRMC 1723:1726 HRMC 1723:1726 HRMC 1723:1726 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 9 9 gly glycopeptides 1675:1687 arg1 cells 1716:1720 human renal mesangial cells 1694:1720 human renal mesangial cells (HRMC) 1694:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 8 10 theme HRP 1460:1462 arg1 digests 1464:1470 HRP digests 1460:1470 HRP digests 1460:1470 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 4 11 theme Fe3O4-DA-Maltose 681:696 arg1 NPs 699:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 0 12 theme Glycopeptides 149:161 arg1 Enrichment 135:144 Highly Selective Enrichment 118:144 Highly Selective Enrichment of Glycopeptides 118:161 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 5 13 theme NPs 850:852 arg1 structure 803:811 structure 803:811 structure 803:811 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 13 theme NPs 850:852 arg1 composition 818:828 composition 818:828 composition 818:828 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 13 theme NPs 850:852 arg1 morphology 791:800 The morphology 787:800 The morphology 787:800 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 1 14 theme biological 246:255 arg1 samples 257:263 biological samples 246:263 biological samples 246:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 4 15 theme -catalyzed 730:739 arg1 chemistry 775:783 click chemistry 769:783 click chemistry 769:783 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 4 15 theme -catalyzed 730:739 arg1 cycloaddition 754:766 copper(I)-catalyzed azide-alkyne cycloaddition 721:766 copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry) 721:784 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 3 16 with chelation 459:467 arg1 atoms 502:506 iron atoms 497:506 iron atoms 497:506 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 5 17 theme vibrating 1040:1048 arg1 magnetometer 1057:1068 vibrating sample magnetometer 1040:1068 vibrating sample magnetometer (VSM) 1040:1074 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 17 theme vibrating 1040:1048 arg1 VSM 1071:1073 VSM 1071:1073 VSM 1071:1073 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 7 18 from isolation 1226:1234 arg1 digests 1324:1330 immunoglobulin (IgG) digests 1303:1330 immunoglobulin (IgG) digests 1303:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 18 from isolation 1226:1234 arg1 HRP 1297:1299 HRP 1297:1299 HRP 1297:1299 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 18 from isolation 1226:1234 arg1 peroxidase 1285:1294 horseradish peroxidase 1273:1294 horseradish peroxidase (HRP) 1273:1300 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 10 19 gly glycosylation 1873:1885 arg2 sites 1887:1891 124 glycosylation sites 1869:1891 124 glycosylation sites 1869:1891 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 10 19 gly glycosylation 1873:1885 arg2 124 1869:1871 124 1869:1871 124 1869:1871 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 7 20 from enrichment 1240:1249 arg1 digests 1324:1330 immunoglobulin (IgG) digests 1303:1330 immunoglobulin (IgG) digests 1303:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 20 from enrichment 1240:1249 arg1 HRP 1297:1299 HRP 1297:1299 HRP 1297:1299 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 20 from enrichment 1240:1249 arg1 peroxidase 1285:1294 horseradish peroxidase 1273:1294 horseradish peroxidase (HRP) 1273:1300 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 4 21 theme copper 721:726 arg1 chemistry 775:783 click chemistry 769:783 click chemistry 769:783 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 4 21 theme copper 721:726 arg1 cycloaddition 754:766 copper(I)-catalyzed azide-alkyne cycloaddition 721:766 copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry) 721:784 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 0 22 theme Nanoparticles 74:86 arg1 Synthesis 6:14 Click Synthesis 0:14 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 0:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 8 23 dep glycopeptides 1581:1593 arg1 enrichment 1595:1604 enrichment 1595:1604 glycopeptides enrichment (85-110%) 1581:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 23 dep glycopeptides 1581:1593 arg1 % 1613:1613 85-110% 1607:1613 85-110% 1607:1613 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 9 24 theme renal 1700:1704 arg1 HRMC 1723:1726 HRMC 1723:1726 HRMC 1723:1726 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 9 24 theme renal 1700:1704 arg1 cells 1716:1720 human renal mesangial cells 1694:1720 human renal mesangial cells (HRMC) 1694:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 8 25 theme ng 1504:1505 arg1 -1 1510:1511 -1 1510:1511 -1 1510:1511 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 25 theme ng 1504:1505 arg1 μL 1507:1508 0.05 ng μL 1499:1508 0.05 ng μL(-1) 1499:1512 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 5 26 theme electron 937:944 arg1 TEM 958:960 TEM 958:960 TEM 958:960 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 26 theme electron 937:944 arg1 microscopy 946:955 transmission electron microscopy 924:955 transmission electron microscopy (TEM) 924:961 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 8 27 theme mass 1347:1350 arg1 analysis 1366:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 9 28 gly N-glycosylation 1751:1765 arg2 sites 1767:1771 N-glycosylation sites 1751:1771 N-glycosylation sites 1751:1771 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 10 29 link N-linked 1811:1818 arg1 glycopeptides 1820:1832 115 different N-linked glycopeptides 1797:1832 115 different N-linked glycopeptides 1797:1832 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 4 30 theme click 769:773 arg1 chemistry 775:783 click chemistry 769:783 click chemistry 769:783 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 4 30 theme click 769:773 arg1 cycloaddition 754:766 copper(I)-catalyzed azide-alkyne cycloaddition 721:766 copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry) 721:784 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 5 31 theme Fe3O4-DA-Maltose 833:848 arg1 NPs 850:852 Fe3O4-DA-Maltose NPs 833:852 Fe3O4-DA-Maltose NPs 833:852 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 32 theme X-ray 964:968 arg1 XRD 990:992 XRD 990:992 XRD 990:992 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 32 theme X-ray 964:968 arg1 diffraction 977:987 X-ray powder diffraction 964:987 X-ray powder diffraction (XRD) 964:993 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 8 33 theme binding 1523:1529 arg1 capacity 1531:1538 a large binding capacity 1515:1538 a large binding capacity up to 43 mg g(-1) 1515:1556 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 34 theme novel 1394:1398 arg1 NPs 1400:1402 the novel NPs 1390:1402 the novel NPs 1390:1402 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 9 35 theme sites 1767:1771 arg1 identification 1733:1746 identification 1733:1746 identification of N-glycosylation sites 1733:1771 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 7 36 theme hydrophilic 1177:1187 arg1 NPs 1206:1208 The hydrophilic Fe3O4-DA-Maltose NPs 1173:1208 The hydrophilic Fe3O4-DA-Maltose NPs 1173:1208 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 3 37 theme catechol 476:483 arg1 ligand 485:490 the catechol ligand 472:490 the catechol ligand 472:490 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 0 38 theme Maltose-Functionalized 31:52 arg1 Nanoparticles 74:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 9 39 from cells 1716:1720 arg1 glycopeptides 1675:1687 glycopeptides 1675:1687 glycopeptides from human renal mesangial cells (HRMC) 1675:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 7 40 theme immunoglobulin 1303:1316 arg1 digests 1324:1330 immunoglobulin (IgG) digests 1303:1330 immunoglobulin (IgG) digests 1303:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 40 theme immunoglobulin 1303:1316 arg1 peroxidase 1285:1294 horseradish peroxidase 1273:1294 horseradish peroxidase (HRP) 1273:1300 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 8 41 theme 43 1546:1547 arg1 mg 1549:1550 mg 1549:1550 mg 1549:1550 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 42 theme detection 1419:1427 arg1 sensitivity 1429:1439 high detection sensitivity 1414:1439 high detection sensitivity 1414:1439 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 3 43 theme azido-terminated 509:524 arg1 DA 536:537 DA 536:537 DA 536:537 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 3 43 theme azido-terminated 509:524 arg1 dopamine 526:533 azido-terminated dopamine 509:533 azido-terminated dopamine (DA) derivative 509:549 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 0 44 theme Oxide 59:63 arg1 Nanoparticles 74:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 5 45 dep Fourier 875:881 arg1 transform 883:891 transform 883:891 transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM) 883:1074 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 46 theme photoelectron 1002:1014 arg1 spectrometer 1016:1027 X-ray photoelectron spectrometer 996:1027 X-ray photoelectron spectrometer (XPS) 996:1033 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 46 theme photoelectron 1002:1014 arg1 XPS 1030:1032 XPS 1030:1032 XPS 1030:1032 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 6 47 theme obtained 1110:1117 arg1 NPs 1119:1121 the obtained NPs 1106:1121 the obtained NPs 1106:1121 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 10 48 theme N-linked 1811:1818 arg1 glycopeptides 1820:1832 115 different N-linked glycopeptides 1797:1832 115 different N-linked glycopeptides 1797:1832 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 8 49 gly glycopeptides 1581:1593 arg2 glycopeptides 1581:1593 glycopeptides enrichment (85-110%) 1581:1614 glycopeptides enrichment (85-110%) 1581:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 7 50 theme horseradish 1273:1283 arg1 digests 1324:1330 immunoglobulin (IgG) digests 1303:1330 immunoglobulin (IgG) digests 1303:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 50 theme horseradish 1273:1283 arg1 HRP 1297:1299 HRP 1297:1299 HRP 1297:1299 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 50 theme horseradish 1273:1283 arg1 peroxidase 1285:1294 horseradish peroxidase 1273:1294 horseradish peroxidase (HRP) 1273:1300 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 8 51 from digests 1464:1470 arg1 enrichment 1444:1453 enrichment 1444:1453 enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%) 1444:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 2 52 theme functionalized 397:410 arg1 NPs 433:435 NPs 433:435 NPs 433:435 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 52 theme functionalized 397:410 arg1 nanoparticles 418:430 hydrophilic maltose functionalized Fe3O4 nanoparticles 377:430 hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 377:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 7 53 theme glycopeptides 1254:1266 arg1 isolation 1226:1234 isolation 1226:1234 isolation 1226:1234 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 53 theme glycopeptides 1254:1266 arg1 enrichment 1240:1249 enrichment 1240:1249 enrichment 1240:1249 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 8 54 theme good 1563:1566 arg1 recovery 1568:1575 good recovery 1563:1575 good recovery for glycopeptides enrichment (85-110%) 1563:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 2 55 theme easy 318:321 arg1 strategy 357:364 an easy and one-step surface modification strategy 315:364 an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 315:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 56 theme modification 344:355 arg1 strategy 357:364 an easy and one-step surface modification strategy 315:364 an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 315:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 57 theme hydrophilic 377:387 arg1 NPs 433:435 NPs 433:435 NPs 433:435 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 57 theme hydrophilic 377:387 arg1 nanoparticles 418:430 hydrophilic maltose functionalized Fe3O4 nanoparticles 377:430 hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 377:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 1 58 gly glycopeptides 227:239 arg2 glycopeptides 227:239 low-abundance glycopeptides 213:239 low-abundance glycopeptides from biological samples 213:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 1 58 gly glycopeptides 227:239 arg1 samples 257:263 biological samples 246:263 biological samples 246:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 5 59 theme infrared 893:900 arg1 spectroscopy 902:913 infrared spectroscopy 893:913 infrared spectroscopy (FT-IR) 893:921 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 59 theme infrared 893:900 arg1 FT-IR 916:920 FT-IR 916:920 FT-IR 916:920 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 7 60 from peroxidase 1285:1294 arg1 isolation 1226:1234 isolation 1226:1234 isolation 1226:1234 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 60 from peroxidase 1285:1294 arg1 glycopeptides 1254:1266 glycopeptides 1254:1266 glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests 1254:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 60 from peroxidase 1285:1294 arg1 enrichment 1240:1249 enrichment 1240:1249 enrichment 1240:1249 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 3 61 theme magnetic 583:590 arg1 nanoparticles 598:610 magnetic Fe3O4 nanoparticles 583:610 magnetic Fe3O4 nanoparticles 583:610 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 6 62 theme contact 1146:1152 arg1 measurement 1160:1170 water contact angle measurement 1140:1170 water contact angle measurement 1140:1170 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 2 63 theme one-step 327:334 arg1 strategy 357:364 an easy and one-step surface modification strategy 315:364 an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 315:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 1 64 theme low-abundance 213:225 arg1 glycopeptides 227:239 low-abundance glycopeptides 213:239 low-abundance glycopeptides from biological samples 213:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 3 65 theme Fe3O4 592:596 arg1 nanoparticles 598:610 magnetic Fe3O4 nanoparticles 583:610 magnetic Fe3O4 nanoparticles 583:610 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 9 66 theme Fe3O4-DA-Maltose 1631:1646 arg1 NPs 1648:1650 the Fe3O4-DA-Maltose NPs 1627:1650 the Fe3O4-DA-Maltose NPs 1627:1650 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 1 67 theme methods 183:189 arg1 crucial 268:274 crucial 268:274 crucial 268:274 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 1 67 theme methods 183:189 arg1 development 168:178 The development 164:178 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples 164:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 4 68 theme maltose-functionalized 651:672 arg1 NPs 699:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 8 69 dep g 1552:1552 arg1 up 1540:1541 up 1540:1541 up 1540:1541 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 3 70 theme nanoparticles 598:610 arg1 surface 572:578 the surface 568:578 the surface of magnetic Fe3O4 nanoparticles 568:610 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 8 71 theme low 1492:1494 arg1 concentration 1475:1487 concentration 1475:1487 concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%) 1475:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 1 72 from samples 257:263 arg1 glycopeptides 227:239 low-abundance glycopeptides 213:239 low-abundance glycopeptides from biological samples 213:263 The development of methods to isolate and enrich low-abundance glycopeptides from biological samples is crucial to glycoproteomics. 26479949 5 73 theme sample 1050:1055 arg1 magnetometer 1057:1068 vibrating sample magnetometer 1040:1068 vibrating sample magnetometer (VSM) 1040:1074 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 73 theme sample 1050:1055 arg1 VSM 1071:1073 VSM 1071:1073 VSM 1071:1073 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 8 74 theme 0.05 1499:1502 arg1 ng 1504:1505 ng 1504:1505 ng 1504:1505 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 4 75 theme azide-alkyne 741:752 arg1 chemistry 775:783 click chemistry 769:783 click chemistry 769:783 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 4 75 theme azide-alkyne 741:752 arg1 cycloaddition 754:766 copper(I)-catalyzed azide-alkyne cycloaddition 721:766 copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry) 721:784 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 8 76 from concentration 1475:1487 arg1 enrichment 1444:1453 enrichment 1444:1453 enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%) 1444:1614 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 9 77 theme human 1694:1698 arg1 HRMC 1723:1726 HRMC 1723:1726 HRMC 1723:1726 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 9 77 theme human 1694:1698 arg1 cells 1716:1720 human renal mesangial cells 1694:1720 human renal mesangial cells (HRMC) 1694:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 0 78 theme Dopamine 97:104 arg1 Anchors 106:112 Dopamine Anchors 97:112 Dopamine Anchors for Highly Selective Enrichment of Glycopeptides 97:161 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 5 79 theme transmission 924:935 arg1 TEM 958:960 TEM 958:960 TEM 958:960 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 79 theme transmission 924:935 arg1 microscopy 946:955 transmission electron microscopy 924:955 transmission electron microscopy (TEM) 924:961 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 9 80 theme mesangial 1706:1714 arg1 HRMC 1723:1726 HRMC 1723:1726 HRMC 1723:1726 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 9 80 theme mesangial 1706:1714 arg1 cells 1716:1720 human renal mesangial cells 1694:1720 human renal mesangial cells (HRMC) 1694:1727 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 8 81 theme MALDI-TOF 1337:1345 arg1 analysis 1366:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 0 82 theme Selective 125:133 arg1 Enrichment 135:144 Highly Selective Enrichment 118:144 Highly Selective Enrichment of Glycopeptides 118:161 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 8 83 theme large 1517:1521 arg1 capacity 1531:1538 a large binding capacity 1515:1538 a large binding capacity up to 43 mg g(-1) 1515:1556 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 84 theme spectrometric 1352:1364 arg1 analysis 1366:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis 1333:1373 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 7 85 theme Fe3O4-DA-Maltose 1189:1204 arg1 NPs 1206:1208 The hydrophilic Fe3O4-DA-Maltose NPs 1173:1208 The hydrophilic Fe3O4-DA-Maltose NPs 1173:1208 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 0 86 theme Hydrophilic 19:29 arg1 Nanoparticles 74:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 5 87 theme powder 970:975 arg1 XRD 990:992 XRD 990:992 XRD 990:992 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 87 theme powder 970:975 arg1 diffraction 977:987 X-ray powder diffraction 964:987 X-ray powder diffraction (XRD) 964:993 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 9 88 theme N-glycosylation 1751:1765 arg1 sites 1767:1771 N-glycosylation sites 1751:1771 N-glycosylation sites 1751:1771 Moreover, the Fe3O4-DA-Maltose NPs were applied to enrich glycopeptides from human renal mesangial cells (HRMC) for identification of N-glycosylation sites. 26479949 10 89 theme different 1801:1809 arg1 glycopeptides 1820:1832 115 different N-linked glycopeptides 1797:1832 115 different N-linked glycopeptides 1797:1832 Finally, we identified 115 different N-linked glycopeptides, representing 93 gene products and 124 glycosylation sites in HRMC. 26479949 3 90 theme iron 497:500 arg1 atoms 502:506 iron atoms 497:506 iron atoms 497:506 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 0 91 theme Iron 54:57 arg1 Nanoparticles 74:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 7 92 gly glycopeptides 1254:1266 arg1 digests 1324:1330 immunoglobulin (IgG) digests 1303:1330 immunoglobulin (IgG) digests 1303:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 92 gly glycopeptides 1254:1266 arg1 HRP 1297:1299 HRP 1297:1299 HRP 1297:1299 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 92 gly glycopeptides 1254:1266 arg1 peroxidase 1285:1294 horseradish peroxidase 1273:1294 horseradish peroxidase (HRP) 1273:1300 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 7 92 gly glycopeptides 1254:1266 arg2 glycopeptides 1254:1266 glycopeptides 1254:1266 glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests 1254:1330 The hydrophilic Fe3O4-DA-Maltose NPs were applied in isolation and enrichment of glycopeptides from horseradish peroxidase (HRP), immunoglobulin (IgG) digests. 26479949 8 93 theme mg 1549:1550 arg1 -1 1554:1555 -1 1554:1555 -1 1554:1555 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 8 93 theme mg 1549:1550 arg1 g 1552:1552 43 mg g 1546:1552 43 mg g(-1) 1546:1556 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 3 94 theme ligand 485:490 arg1 chelation 459:467 the chelation 455:467 the chelation of the catechol ligand with iron atoms 455:506 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 0 95 theme Magnetic 65:72 arg1 Nanoparticles 74:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles 19:86 Click Synthesis of Hydrophilic Maltose-Functionalized Iron Oxide Magnetic Nanoparticles Based on Dopamine Anchors for Highly Selective Enrichment of Glycopeptides. 26479949 8 96 theme high 1414:1417 arg1 sensitivity 1429:1439 high detection sensitivity 1414:1439 high detection sensitivity 1414:1439 The MALDI-TOF mass spectrometric analysis indicated that the novel NPs exhibited high detection sensitivity in enrichment from HRP digests at concentration as low as 0.05 ng μL(-1), a large binding capacity up to 43 mg g(-1), and good recovery for glycopeptides enrichment (85-110%). 26479949 3 97 theme dopamine 526:533 arg1 derivative 540:549 azido-terminated dopamine (DA) derivative 509:549 azido-terminated dopamine (DA) derivative 509:549 First, based on the chelation of the catechol ligand with iron atoms, azido-terminated dopamine (DA) derivative was assembled on the surface of magnetic Fe3O4 nanoparticles by sonication. 26479949 5 98 theme X-ray 996:1000 arg1 spectrometer 1016:1027 X-ray photoelectron spectrometer 996:1027 X-ray photoelectron spectrometer (XPS) 996:1033 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 5 98 theme X-ray 996:1000 arg1 XPS 1030:1032 XPS 1030:1032 XPS 1030:1032 The morphology, structure, and composition of Fe3O4-DA-Maltose NPs were investigated by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectrometer (XPS), and vibrating sample magnetometer (VSM). 26479949 6 99 theme NPs 1119:1121 arg1 hydrophilicity 1088:1101 hydrophilicity 1088:1101 hydrophilicity of the obtained NPs 1088:1121 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 6 99 theme NPs 1119:1121 arg1 Meanwhile 1077:1085 Meanwhile 1077:1085 Meanwhile 1077:1085 Meanwhile, hydrophilicity of the obtained NPs was evaluated by water contact angle measurement. 26479949 4 100 theme hydrophilic 639:649 arg1 NPs 699:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs 635:701 Second, the hydrophilic maltose-functionalized Fe3O4 (Fe3O4-DA-Maltose) NPs were obtained via copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry). 26479949 2 101 theme Fe3O4 412:416 arg1 NPs 433:435 NPs 433:435 NPs 433:435 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 26479949 2 101 theme Fe3O4 412:416 arg1 nanoparticles 418:430 hydrophilic maltose functionalized Fe3O4 nanoparticles 377:430 hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs) 377:436 Herein, we present an easy and one-step surface modification strategy to prepare hydrophilic maltose functionalized Fe3O4 nanoparticles (NPs). 25568322 0 0 link prostate-derived 84:99 arg1 protein 117:123 a prostate-derived heparin-binding protein WGA16 82:129 a prostate-derived heparin-binding protein WGA16 from boar sperm 82:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 3 1 theme plasma 493:498 arg1 WGA16 457:461 WGA16 457:461 WGA16 457:461 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 3 1 theme plasma 493:498 arg1 protein 500:506 a prostate-derived seminal plasma protein 466:506 a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract 466:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 6 2 theme plasma 1124:1129 arg1 proteins 1131:1138 seminal plasma proteins 1116:1138 seminal plasma proteins 1116:1138 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 4 3 theme cDNA 628:631 arg1 sequences 633:641 protein and cDNA sequences 616:641 sequences 633:641 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 10 4 theme N-glycosylated 1508:1521 arg1 sites 1523:1527 N-glycosylated sites 1508:1527 N-glycosylated sites 1508:1527 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 11 5 theme sites 1620:1624 arg1 geography 1592:1600 This geography 1587:1600 This geography of two functional sites 1587:1624 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 6 6 theme N-glycosylation 1086:1100 arg1 profile 1102:1108 its unique N-glycosylation profile 1075:1108 its unique N-glycosylation profile among seminal plasma proteins 1075:1138 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 11 7 theme heparin-binding 1772:1786 arg1 domain 1788:1793 its heparin-binding domain 1768:1793 its heparin-binding domain 1768:1793 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 1 8 theme maturation 203:212 arg1 process 214:220 a functional maturation process 190:220 a functional maturation process 190:220 Mammalian sperm acquire fertility through a functional maturation process called capacitation, where sperm membrane molecules are drastically remodeled. 25568322 10 9 theme potential 1535:1543 arg1 site 1561:1564 a potential heparin-binding site 1533:1564 a potential heparin-binding site 1533:1564 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 9 10 theme WGA16 1420:1424 arg1 removal 1409:1415 surface removal 1401:1415 surface removal of WGA16 1401:1424 Interestingly, surface removal of WGA16 is experimentally induced by either UDP-Gal or heparin. 25568322 4 11 theme protein 616:622 arg1 sequences 633:641 protein and cDNA sequences 616:641 sequences 633:641 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 10 12 theme opposite 1571:1578 arg1 sides 1580:1584 opposite sides 1571:1584 opposite sides 1571:1584 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 3 13 theme sperm 561:565 arg1 surface 567:573 the sperm surface 557:573 the sperm surface 557:573 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 4 14 theme purified 647:654 arg1 WGA16 656:660 purified WGA16 647:660 purified WGA16 647:660 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 7 15 theme GalNAc 1167:1172 arg1 residues 1174:1181 exposed GlcNAc and GalNAc residues 1148:1181 exposed GlcNAc and GalNAc residues 1148:1181 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 3 16 theme male 582:585 arg1 tract 600:604 the male reproductive tract 578:604 the male reproductive tract 578:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 7 17 theme GlcNAc 1156:1161 arg1 residues 1174:1181 exposed GlcNAc and GalNAc residues 1148:1181 exposed GlcNAc and GalNAc residues 1148:1181 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 0 18 from Discovery 0:8 arg1 sperm 141:145 boar sperm 136:145 boar sperm 136:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 8 19 theme surface 1283:1289 arg1 galactosyltransferase 1291:1311 a sperm surface galactosyltransferase 1275:1311 a sperm surface galactosyltransferase 1275:1311 The GlcNAc/GalNAc residues can work as binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal. 25568322 0 20 theme prostate-derived 84:99 arg1 protein 117:123 a prostate-derived heparin-binding protein WGA16 82:129 a prostate-derived heparin-binding protein WGA16 from boar sperm 82:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 21 from structures 32:41 arg1 sperm 141:145 boar sperm 136:145 boar sperm 136:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 22 theme protein 117:123 arg1 Discovery 0:8 Discovery 0:8 Discovery 0:8 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 22 theme protein 117:123 arg1 structures 32:41 crystal structures 24:41 crystal structures 24:41 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 22 theme protein 117:123 arg1 properties 68:77 capacitation-related properties 47:77 capacitation-related properties 47:77 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 2 23 theme sperm 426:430 arg1 surface 432:438 the sperm surface 422:438 the sperm surface on capacitation 422:454 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 4 24 theme zymogen 690:696 arg1 protein 706:712 human zymogen granule protein 16 684:715 human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures 684:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 4 24 theme zymogen 690:696 arg1 ZG16 718:721 ZG16 718:721 ZG16 718:721 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 2 25 theme lipid 380:384 arg1 rafts 386:390 lipid rafts 380:390 lipid rafts 380:390 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 2 26 attach removed 409:415 arg2 protein 369:375 a wheat germ agglutinin (WGA)-reactive protein 330:375 a wheat germ agglutinin (WGA)-reactive protein 330:375 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 2 26 attach removed 409:415 arg1 surface 432:438 the sperm surface 422:438 the sperm surface on capacitation 422:454 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 5 27 theme basic 868:872 arg1 patch 874:878 a basic patch 866:878 a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 866:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 11 28 from glycosaminoglycans 1834:1851 arg1 tract 1867:1871 the female tract 1856:1871 the female tract 1856:1871 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 4 29 theme protein 706:712 arg1 it 663:664 it 663:664 it 663:664 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 4 29 theme protein 706:712 arg1 homologue 671:679 a homologue 669:679 a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures 669:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 1 30 theme Mammalian 148:156 arg1 sperm 158:162 Mammalian sperm 148:162 Mammalian sperm 148:162 Mammalian sperm acquire fertility through a functional maturation process called capacitation, where sperm membrane molecules are drastically remodeled. 25568322 11 31 theme WGA16 1894:1898 arg1 removal 1883:1889 removal 1883:1889 removal of WGA16 from the sperm surface 1883:1921 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 0 32 theme capacitation-related 47:66 arg1 properties 68:77 capacitation-related properties 47:77 capacitation-related properties 47:77 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 11 33 theme sulfated 1825:1832 arg1 glycosaminoglycans 1834:1851 sulfated glycosaminoglycans 1825:1851 sulfated glycosaminoglycans 1825:1851 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 5 34 theme family 964:969 arg1 domain 946:951 the conventional lectin domain 922:951 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 34 theme family 964:969 arg1 family 964:969 the JRL family 956:969 the JRL family 956:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 2 35 theme germ 338:341 arg1 agglutinin 343:352 wheat germ agglutinin 332:352 wheat germ agglutinin (WGA) 332:358 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 2 35 theme germ 338:341 arg1 WGA 355:357 WGA 355:357 WGA 355:357 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 5 36 theme conventional 926:937 arg1 domain 946:951 the conventional lectin domain 922:951 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 36 theme conventional 926:937 arg1 family 964:969 the JRL family 956:969 the JRL family 956:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 6 37 theme comparative 1031:1041 arg1 spectrometry 1048:1059 comparative mass spectrometry 1031:1059 comparative mass spectrometry 1031:1059 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 0 38 from sperm 141:145 arg1 protein 117:123 a prostate-derived heparin-binding protein WGA16 82:129 a prostate-derived heparin-binding protein WGA16 from boar sperm 82:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 38 from sperm 141:145 arg1 Discovery 0:8 Discovery 0:8 Discovery 0:8 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 38 from sperm 141:145 arg1 structures 32:41 crystal structures 24:41 crystal structures 24:41 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 0 38 from sperm 141:145 arg1 properties 68:77 capacitation-related properties 47:77 capacitation-related properties 47:77 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 4 39 theme Jacalin-related 741:755 arg1 family 770:775 the Jacalin-related lectin (JRL) family 737:775 the Jacalin-related lectin (JRL) family in crystal and primary structures 737:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 11 40 theme female 1860:1865 arg1 tract 1867:1871 the female tract 1856:1871 the female tract 1856:1871 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 1 41 theme membrane 255:262 arg1 molecules 264:272 sperm membrane molecules 249:272 sperm membrane molecules 249:272 Mammalian sperm acquire fertility through a functional maturation process called capacitation, where sperm membrane molecules are drastically remodeled. 25568322 7 42 theme Gal 1202:1204 arg1 residues 1206:1213 additional Gal residues 1191:1213 additional Gal residues 1191:1213 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 10 43 theme crystal 1489:1495 arg1 structure 1497:1505 the crystal structure 1485:1505 the crystal structure 1485:1505 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 11 44 theme functional 1609:1618 arg1 sites 1620:1624 two functional sites 1605:1624 two functional sites 1605:1624 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 3 45 theme seminal 485:491 arg1 WGA16 457:461 WGA16 457:461 WGA16 457:461 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 3 45 theme seminal 485:491 arg1 protein 500:506 a prostate-derived seminal plasma protein 466:506 a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract 466:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 10 46 from face 1566:1569 arg1 structure 1497:1505 the crystal structure 1485:1505 the crystal structure 1485:1505 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 4 47 theme primary 792:798 arg1 structures 800:809 crystal and primary structures 780:809 structures 800:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 6 48 theme ZG16 1013:1016 arg1 members 1018:1024 other ZG16 members 1007:1024 other ZG16 members 1007:1024 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 6 49 theme seminal 1116:1122 arg1 proteins 1131:1138 seminal plasma proteins 1116:1138 seminal plasma proteins 1116:1138 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 9 50 theme surface 1401:1407 arg1 removal 1409:1415 surface removal 1401:1415 surface removal of WGA16 1401:1424 Interestingly, surface removal of WGA16 is experimentally induced by either UDP-Gal or heparin. 25568322 1 51 theme functional 192:201 arg1 process 214:220 a functional maturation process 190:220 a functional maturation process 190:220 Mammalian sperm acquire fertility through a functional maturation process called capacitation, where sperm membrane molecules are drastically remodeled. 25568322 4 52 theme crystal 780:786 arg1 structures 800:809 crystal and primary structures 780:809 structures 800:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 2 53 from protein 369:375 arg1 rafts 386:390 lipid rafts 380:390 lipid rafts 380:390 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 6 54 theme unique 1079:1084 arg1 profile 1102:1108 its unique N-glycosylation profile 1075:1108 its unique N-glycosylation profile among seminal plasma proteins 1075:1138 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 5 55 contain containing 880:889 arg1 patch 874:878 a basic patch 866:878 a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 866:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 55 contain containing 880:889 arg2 family 964:969 the JRL family 956:969 the JRL family 956:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 55 contain containing 880:889 arg2 residues 905:912 Lys-53/Lys-73 residues 891:912 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 55 contain containing 880:889 arg2 domain 946:951 the conventional lectin domain 922:951 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 10 56 theme heparin-binding 1545:1559 arg1 site 1561:1564 a potential heparin-binding site 1533:1564 a potential heparin-binding site 1533:1564 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 0 57 theme boar 136:139 arg1 sperm 141:145 boar sperm 136:145 boar sperm 136:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 11 58 from surface 1915:1921 arg1 removal 1883:1889 removal 1883:1889 removal of WGA16 from the sperm surface 1883:1921 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 0 59 from properties 68:77 arg1 sperm 141:145 boar sperm 136:145 boar sperm 136:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 10 60 gly N-glycosylated 1508:1521 arg1 sites 1523:1527 N-glycosylated sites 1508:1527 N-glycosylated sites 1508:1527 In the crystal structure, N-glycosylated sites and a potential heparin-binding site face opposite sides. 25568322 3 61 theme reproductive 587:598 arg1 tract 600:604 the male reproductive tract 578:604 the male reproductive tract 578:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 3 62 link prostate-derived 468:483 arg1 WGA16 457:461 WGA16 457:461 WGA16 457:461 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 3 62 link prostate-derived 468:483 arg1 protein 500:506 a prostate-derived seminal plasma protein 466:506 a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract 466:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 5 63 theme glycan 814:819 arg1 array 821:825 A glycan array 812:825 A glycan array 812:825 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 8 64 theme binding 1255:1261 arg1 ligands 1263:1269 binding ligands 1255:1269 binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal 1255:1383 The GlcNAc/GalNAc residues can work as binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal. 25568322 7 65 theme exposed 1148:1154 arg1 residues 1174:1181 exposed GlcNAc and GalNAc residues 1148:1181 exposed GlcNAc and GalNAc residues 1148:1181 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 0 66 theme heparin-binding 101:115 arg1 protein 117:123 a prostate-derived heparin-binding protein WGA16 82:129 a prostate-derived heparin-binding protein WGA16 from boar sperm 82:145 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 5 67 theme Lys-53/Lys-73 891:903 arg1 residues 905:912 Lys-53/Lys-73 residues 891:912 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 8 68 theme sperm 1277:1281 arg1 galactosyltransferase 1291:1311 a sperm surface galactosyltransferase 1275:1311 a sperm surface galactosyltransferase 1275:1311 The GlcNAc/GalNAc residues can work as binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal. 25568322 11 69 theme surface 1729:1735 arg1 galactosyltransferase 1737:1757 the surface galactosyltransferase 1725:1757 the surface galactosyltransferase 1725:1757 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 6 70 theme other 1007:1011 arg1 members 1018:1024 other ZG16 members 1007:1024 other ZG16 members 1007:1024 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 4 71 from family 770:775 arg1 structures 800:809 crystal and primary structures 780:809 structures 800:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 2 72 from surface 432:438 arg1 capacitation 443:454 capacitation 443:454 capacitation 443:454 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 4 73 theme human 684:688 arg1 protein 706:712 human zymogen granule protein 16 684:715 human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures 684:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 4 73 theme human 684:688 arg1 ZG16 718:721 ZG16 718:721 ZG16 718:721 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 6 74 gly glycosylated 981:992 arg1 WGA16 972:976 WGA16 972:976 WGA16 972:976 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 4 75 theme granule 698:704 arg1 protein 706:712 human zymogen granule protein 16 684:715 human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures 684:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 4 75 theme granule 698:704 arg1 ZG16 718:721 ZG16 718:721 ZG16 718:721 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 11 76 theme sperm 1909:1913 arg1 surface 1915:1921 the sperm surface 1905:1921 the sperm surface 1905:1921 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 0 77 theme crystal 24:30 arg1 structures 32:41 crystal structures 24:41 crystal structures 24:41 Discovery, primary, and crystal structures and capacitation-related properties of a prostate-derived heparin-binding protein WGA16 from boar sperm. 25568322 8 78 theme GlcNAc/GalNAc 1220:1232 arg1 residues 1234:1241 The GlcNAc/GalNAc residues 1216:1241 The GlcNAc/GalNAc residues 1216:1241 The GlcNAc/GalNAc residues can work as binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal. 25568322 4 79 theme lectin 757:762 arg1 family 770:775 the Jacalin-related lectin (JRL) family 737:775 the Jacalin-related lectin (JRL) family in crystal and primary structures 737:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 5 80 theme JRL 960:962 arg1 family 964:969 the JRL family 956:969 the JRL family 956:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 4 81 theme JRL 765:767 arg1 family 770:775 the Jacalin-related lectin (JRL) family 737:775 the Jacalin-related lectin (JRL) family in crystal and primary structures 737:809 Based on protein and cDNA sequences for purified WGA16, it is a homologue of human zymogen granule protein 16 (ZG16) belonging to the Jacalin-related lectin (JRL) family in crystal and primary structures. 25568322 7 82 contain has 1144:1146 arg1 It 1141:1142 It 1141:1142 It 1141:1142 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 7 82 contain has 1144:1146 arg2 residues 1174:1181 exposed GlcNAc and GalNAc residues 1148:1181 exposed GlcNAc and GalNAc residues 1148:1181 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 3 83 theme prostate-derived 468:483 arg1 WGA16 457:461 WGA16 457:461 WGA16 457:461 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 3 83 theme prostate-derived 468:483 arg1 protein 500:506 a prostate-derived seminal plasma protein 466:506 a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract 466:604 WGA16 is a prostate-derived seminal plasma protein that has never been reported and is deposited on the sperm surface in the male reproductive tract. 25568322 6 84 theme mass 1043:1046 arg1 spectrometry 1048:1059 comparative mass spectrometry 1031:1059 comparative mass spectrometry 1031:1059 WGA16 is glycosylated, contrary to other ZG16 members, and comparative mass spectrometry clearly shows its unique N-glycosylation profile among seminal plasma proteins. 25568322 11 85 theme sperm 1665:1669 arg1 surface 1671:1677 the sperm surface 1661:1677 the sperm surface 1661:1677 This geography of two functional sites suggest that WGA16 is deposited on the sperm surface through interaction between its N-glycans and the surface galactosyltransferase, whereas its heparin-binding domain may be involved in binding to sulfated glycosaminoglycans in the female tract, enabling removal of WGA16 from the sperm surface. 25568322 2 86 theme wheat 332:336 arg1 agglutinin 343:352 wheat germ agglutinin 332:352 wheat germ agglutinin (WGA) 332:358 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 2 86 theme wheat 332:336 arg1 WGA 355:357 WGA 355:357 WGA 355:357 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 25568322 1 87 theme sperm 249:253 arg1 molecules 264:272 sperm membrane molecules 249:272 sperm membrane molecules 249:272 Mammalian sperm acquire fertility through a functional maturation process called capacitation, where sperm membrane molecules are drastically remodeled. 25568322 5 88 theme lectin 939:944 arg1 domain 946:951 the conventional lectin domain 922:951 Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family 891:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 5 88 theme lectin 939:944 arg1 family 964:969 the JRL family 956:969 the JRL family 956:969 A glycan array shows that WGA16 binds heparin through a basic patch containing Lys-53/Lys-73 residues but not the conventional lectin domain of the JRL family. 25568322 7 89 theme additional 1191:1200 arg1 residues 1206:1213 additional Gal residues 1191:1213 additional Gal residues 1191:1213 It has exposed GlcNAc and GalNAc residues without additional Gal residues. 25568322 8 90 theme UDP-Gal 1377:1383 arg1 presence 1365:1372 the presence 1361:1372 the presence of UDP-Gal 1361:1383 The GlcNAc/GalNAc residues can work as binding ligands for a sperm surface galactosyltransferase, which actually galactosylates WGA16 in situ in the presence of UDP-Gal. 25568322 2 91 theme -reactive 359:367 arg1 protein 369:375 a wheat germ agglutinin (WGA)-reactive protein 330:375 a wheat germ agglutinin (WGA)-reactive protein 330:375 In this study, we found that a wheat germ agglutinin (WGA)-reactive protein on lipid rafts, named WGA16, is removed from the sperm surface on capacitation. 29106908 0 0 theme step 70:73 arg1 chromatography 84:97 highly efficient single step affinity chromatography 46:97 highly efficient single step affinity chromatography 46:97 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 29106908 3 1 from range 425:429 arg1 stable 405:410 stable 405:410 stable 405:410 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 5 2 theme binding 736:742 arg1 capacity 744:751 retained antigen binding capacity 719:751 retained antigen binding capacity 719:751 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 10 3 theme detailed 1178:1185 arg1 analysis 1194:1201 the first detailed glycan analysis 1168:1201 the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM 1168:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 0 4 theme single 63:68 arg1 chromatography 84:97 highly efficient single step affinity chromatography 46:97 highly efficient single step affinity chromatography 46:97 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 29106908 10 5 theme first 1172:1176 arg1 analysis 1194:1201 the first detailed glycan analysis 1168:1201 the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM 1168:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 3 6 from stable 405:410 arg1 range 425:429 a limited range 415:429 a limited range of conditions 415:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 5 7 theme single 653:658 arg1 strategy 678:685 our single step purification strategy 649:685 our single step purification strategy 649:685 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 10 8 theme similar 1356:1362 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 9 9 theme CHO 1145:1147 arg1 cells 1150:1154 Chinese hamster ovary (CHO) cells 1122:1154 Chinese hamster ovary (CHO) cells 1122:1154 The herein used IgMs were produced in Chinese hamster ovary (CHO) cells. 29106908 3 10 theme complex 350:356 arg1 proteins 382:389 large, complex and highly glycosylated proteins 343:389 large, complex and highly glycosylated proteins that are only stable in a limited range of conditions 343:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 3 10 theme complex 350:356 arg1 IgMs 334:337 IgMs 334:337 IgMs 334:337 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 6 11 theme affinity 872:879 arg1 chromatography 881:894 affinity chromatography 872:894 affinity chromatography 872:894 Here we show that IgMs are sensitive to aggregation at very acidic conditions (pH ≤ 3.0) despite often being used for affinity chromatography. 29106908 5 12 theme step 660:663 arg1 strategy 678:685 our single step purification strategy 649:685 our single step purification strategy 649:685 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 7 13 theme high 923:926 arg1 concentration 933:945 a high salt concentration 921:945 a high salt concentration 921:945 We combined pH 3.5 with a high salt concentration to prevent aggregation during elution. 29106908 10 14 theme type 1259:1262 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 5 15 theme antigen 728:734 arg1 capacity 744:751 retained antigen binding capacity 719:751 retained antigen binding capacity 719:751 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 10 16 from Asn563 1349:1354 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 16 from Asn563 1349:1354 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 5 17 theme screening 618:626 arg1 system 628:633 a small-scale screening system 604:633 a small-scale screening system 604:633 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 5 18 theme small-scale 606:616 arg1 system 628:633 a small-scale screening system 604:633 a small-scale screening system 604:633 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 10 19 theme complex 1251:1257 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 1 20 theme promising 150:158 arg1 tools 160:164 promising tools 150:164 promising tools for therapy and diagnostic approaches 150:202 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 1 20 theme promising 150:158 arg1 antibodies 123:132 Immunoglobulin M (IgM) antibodies 100:132 Immunoglobulin M (IgM) antibodies 100:132 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 3 21 theme large 343:347 arg1 proteins 382:389 large, complex and highly glycosylated proteins 343:389 large, complex and highly glycosylated proteins that are only stable in a limited range of conditions 343:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 3 21 theme large 343:347 arg1 IgMs 334:337 IgMs 334:337 IgMs 334:337 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 4 22 theme IgM 477:479 arg1 antibodies 481:490 these sensitive IgM antibodies 461:490 these sensitive IgM antibodies 461:490 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 3 23 gly glycosylated 369:380 arg1 proteins 382:389 large, complex and highly glycosylated proteins 343:389 large, complex and highly glycosylated proteins that are only stable in a limited range of conditions 343:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 3 23 gly glycosylated 369:380 arg1 IgMs 334:337 IgMs 334:337 IgMs 334:337 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 10 24 theme oligomannosidic 1308:1322 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 25 from Asn332 1286:1291 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 25 from Asn332 1286:1291 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 3 26 theme glycosylated 369:380 arg1 proteins 382:389 large, complex and highly glycosylated proteins 343:389 large, complex and highly glycosylated proteins that are only stable in a limited range of conditions 343:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 3 26 theme glycosylated 369:380 arg1 IgMs 334:337 IgMs 334:337 IgMs 334:337 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 4 27 theme sensitive 467:475 arg1 antibodies 481:490 these sensitive IgM antibodies 461:490 these sensitive IgM antibodies 461:490 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 0 28 theme Glycan 0:5 arg1 profile 7:13 Glycan profile 0:13 Glycan profile of CHO 0:20 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 29106908 4 29 theme available 547:555 arg1 CaptureSelect™ 578:591 CaptureSelect™ 578:591 CaptureSelect™ 578:591 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 4 29 theme available 547:555 arg1 matrix 570:575 a commercially available IgM affinity matrix 532:575 a commercially available IgM affinity matrix (CaptureSelect™) 532:592 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 8 30 theme IgM 1044:1046 arg1 processes 1048:1056 IgM processes 1044:1056 IgM processes for further applications 1044:1081 The elution strategy presented in this paper will improve IgM processes for further applications. 29106908 10 31 from Asn395 1297:1302 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 31 from Asn395 1297:1302 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 0 32 theme CHO 18:20 arg1 profile 7:13 Glycan profile 0:13 Glycan profile of CHO 0:20 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 29106908 4 33 dep optimized 495:503 arg1 investigate 449:459 investigate 449:459 To investigate these sensitive IgM antibodies 446:490 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 7 34 theme salt 928:931 arg1 concentration 933:945 a high salt concentration 921:945 a high salt concentration 921:945 We combined pH 3.5 with a high salt concentration to prevent aggregation during elution. 29106908 2 35 theme recombinant 284:294 arg1 production 296:305 recombinant production 284:305 recombinant production 284:305 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 4 36 theme affinity 561:568 arg1 CaptureSelect™ 578:591 CaptureSelect™ 578:591 CaptureSelect™ 578:591 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 4 36 theme affinity 561:568 arg1 matrix 570:575 a commercially available IgM affinity matrix 532:575 a commercially available IgM affinity matrix (CaptureSelect™) 532:592 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 1 37 theme Immunoglobulin 100:113 arg1 IgM 118:120 IgM 118:120 IgM 118:120 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 1 37 theme Immunoglobulin 100:113 arg1 M 115:115 Immunoglobulin M 100:115 Immunoglobulin M (IgM) antibodies 100:132 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 9 38 theme used 1095:1098 arg1 IgMs 1100:1103 The herein used IgMs 1084:1103 The herein used IgMs 1084:1103 The herein used IgMs were produced in Chinese hamster ovary (CHO) cells. 29106908 1 39 theme diagnostic 182:191 arg1 approaches 193:202 diagnostic approaches 182:202 diagnostic approaches 182:202 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 4 40 theme IgM 557:559 arg1 CaptureSelect™ 578:591 CaptureSelect™ 578:591 CaptureSelect™ 578:591 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 4 40 theme IgM 557:559 arg1 matrix 570:575 a commercially available IgM affinity matrix 532:575 a commercially available IgM affinity matrix (CaptureSelect™) 532:592 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 5 41 theme high 704:707 arg1 yield 709:713 high yield 704:713 high yield 704:713 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 2 42 theme commercial 223:232 arg1 success 234:240 the commercial success 219:240 the commercial success of IgMs 219:248 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 10 43 from Asn402 1338:1343 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 43 from Asn402 1338:1343 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 3 44 theme limited 417:423 arg1 range 425:429 a limited range 415:429 a limited range of conditions 415:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 5 45 theme retained 719:726 arg1 capacity 744:751 retained antigen binding capacity 719:751 retained antigen binding capacity 719:751 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 3 46 theme conditions 434:443 arg1 range 425:429 a limited range 415:429 a limited range of conditions 415:443 IgMs are large, complex and highly glycosylated proteins that are only stable in a limited range of conditions. 29106908 4 47 theme elution 509:515 arg1 conditions 517:526 the elution conditions 505:526 the elution conditions 505:526 To investigate these sensitive IgM antibodies we optimized the elution conditions for a commercially available IgM affinity matrix (CaptureSelect™). 29106908 5 48 theme purification 665:676 arg1 strategy 678:685 our single step purification strategy 649:685 our single step purification strategy 649:685 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 1 49 theme M 115:115 arg1 tools 160:164 promising tools 150:164 promising tools for therapy and diagnostic approaches 150:202 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 1 49 theme M 115:115 arg1 antibodies 123:132 Immunoglobulin M (IgM) antibodies 100:132 Immunoglobulin M (IgM) antibodies 100:132 Immunoglobulin M (IgM) antibodies are reckoned as promising tools for therapy and diagnostic approaches. 29106908 6 50 from conditions 821:830 arg1 sensitive 781:789 sensitive 781:789 sensitive 781:789 Here we show that IgMs are sensitive to aggregation at very acidic conditions (pH ≤ 3.0) despite often being used for affinity chromatography. 29106908 8 51 theme further 1062:1068 arg1 applications 1070:1081 further applications 1062:1081 further applications 1062:1081 The elution strategy presented in this paper will improve IgM processes for further applications. 29106908 10 52 theme IgM 1206:1208 arg1 analysis 1194:1201 the first detailed glycan analysis 1168:1201 the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM 1168:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 8 53 theme elution 990:996 arg1 strategy 998:1005 The elution strategy 986:1005 The elution strategy presented in this paper 986:1029 The elution strategy presented in this paper will improve IgM processes for further applications. 29106908 10 54 with cells 1226:1230 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 54 with cells 1226:1230 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 9 55 theme Chinese 1122:1128 arg1 cells 1150:1154 Chinese hamster ovary (CHO) cells 1122:1154 Chinese hamster ovary (CHO) cells 1122:1154 The herein used IgMs were produced in Chinese hamster ovary (CHO) cells. 29106908 5 56 theme high 691:694 arg1 purity 696:701 high purity 691:701 high purity 691:701 Applying a small-scale screening system, we optimized our single step purification strategy for high purity, high yield and retained antigen binding capacity. 29106908 6 57 theme acidic 814:819 arg1 conditions 821:830 very acidic conditions 809:830 very acidic conditions (pH ≤ 3.0) 809:841 Here we show that IgMs are sensitive to aggregation at very acidic conditions (pH ≤ 3.0) despite often being used for affinity chromatography. 29106908 6 57 theme acidic 814:819 arg1 pH ≤ 3.0 833:840 pH ≤ 3.0 833:840 pH ≤ 3.0 833:840 Here we show that IgMs are sensitive to aggregation at very acidic conditions (pH ≤ 3.0) despite often being used for affinity chromatography. 29106908 2 58 theme IgMs 245:248 arg1 success 234:240 the commercial success 219:240 the commercial success of IgMs 219:248 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 9 59 theme hamster 1130:1136 arg1 cells 1150:1154 Chinese hamster ovary (CHO) cells 1122:1154 Chinese hamster ovary (CHO) cells 1122:1154 The herein used IgMs were produced in Chinese hamster ovary (CHO) cells. 29106908 2 60 theme downstream 311:320 arg1 processing 322:331 downstream processing 311:331 downstream processing 311:331 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 2 61 from bottlenecks 269:279 arg1 production 296:305 recombinant production 284:305 recombinant production 284:305 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 2 61 from bottlenecks 269:279 arg1 processing 322:331 downstream processing 311:331 downstream processing 311:331 Nevertheless, the commercial success of IgMs is hampered due to bottlenecks in recombinant production and downstream processing. 29106908 0 62 theme efficient 53:61 arg1 chromatography 84:97 highly efficient single step affinity chromatography 46:97 highly efficient single step affinity chromatography 46:97 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 29106908 9 63 theme ovary 1138:1142 arg1 cells 1150:1154 Chinese hamster ovary (CHO) cells 1122:1154 Chinese hamster ovary (CHO) cells 1122:1154 The herein used IgMs were produced in Chinese hamster ovary (CHO) cells. 29106908 10 64 theme glycan 1187:1192 arg1 analysis 1194:1201 the first detailed glycan analysis 1168:1201 the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM 1168:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 65 theme human 1367:1371 arg1 serum-IgM 1373:1381 human serum-IgM 1367:1381 human serum-IgM 1367:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 66 from Asn171 1278:1283 arg1 structures 1324:1333 oligomannosidic structures 1308:1333 oligomannosidic structures at Asn402 and Asn563 1308:1354 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 66 from Asn171 1278:1283 arg1 structures 1264:1273 predominantly complex type structures 1237:1273 predominantly complex type structures at Asn171, Asn332 and Asn395 1237:1302 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 10 67 theme CHO 1222:1224 arg1 cells 1226:1230 CHO cells 1222:1230 CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM 1222:1381 We present the first detailed glycan analysis of IgM produced in CHO cells with predominantly complex type structures at Asn171, Asn332 and Asn395 and oligomannosidic structures at Asn402 and Asn563 similar to human serum-IgM. 29106908 0 68 theme affinity 75:82 arg1 chromatography 84:97 highly efficient single step affinity chromatography 46:97 highly efficient single step affinity chromatography 46:97 Glycan profile of CHO derived IgM purified by highly efficient single step affinity chromatography. 28378791 3 0 theme molecular 299:307 arg1 simulations 318:328 extensive molecular dynamics simulations 289:328 extensive molecular dynamics simulations of GluN1 289:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 2 1 from effects 182:188 arg1 dynamics 228:235 dynamics 228:235 dynamics 228:235 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 2 1 from effects 182:188 arg1 structure 214:222 structure 214:222 structure 214:222 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 8 2 theme glycans 1113:1119 arg1 role 1105:1108 an intramolecular potentiating role 1074:1108 an intramolecular potentiating role of glycans on NMDA receptors 1074:1137 Overall, our results suggest an intramolecular potentiating role of glycans on NMDA receptors. 28378791 3 3 theme dynamics 309:316 arg1 simulations 318:328 extensive molecular dynamics simulations 289:328 extensive molecular dynamics simulations of GluN1 289:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 7 4 theme glycosylation-preventing 943:966 arg1 mutation 980:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 3 5 theme GluN2B 343:348 arg1 LBDs 374:377 LBDs 374:377 LBDs 374:377 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 5 theme GluN2B 343:348 arg1 NMDARs 383:388 NMDARs 383:388 NMDARs 383:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 5 theme GluN2B 343:348 arg1 domains 365:371 GluN2B ligand binding domains 343:371 GluN2B ligand binding domains (LBDs) of NMDARs 343:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 5 theme GluN2B 343:348 arg1 GluN1 333:337 GluN1 333:337 GluN1 333:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 7 6 theme GluN1-N440Q 968:978 arg1 mutation 980:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 3 7 theme ligand 350:355 arg1 LBDs 374:377 LBDs 374:377 LBDs 374:377 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 7 theme ligand 350:355 arg1 NMDARs 383:388 NMDARs 383:388 NMDARs 383:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 7 theme ligand 350:355 arg1 domains 365:371 GluN2B ligand binding domains 343:371 GluN2B ligand binding domains (LBDs) of NMDARs 343:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 7 theme ligand 350:355 arg1 GluN1 333:337 GluN1 333:337 GluN1 333:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 7 8 theme electrophysiological 884:903 arg1 analysis 905:912 electrophysiological analysis 884:912 electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation 884:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 3 9 theme GluN1 333:337 arg1 simulations 318:328 extensive molecular dynamics simulations 289:328 extensive molecular dynamics simulations of GluN1 289:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 9 theme GluN1 333:337 arg1 LBDs 374:377 LBDs 374:377 LBDs 374:377 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 9 theme GluN1 333:337 arg1 NMDARs 383:388 NMDARs 383:388 NMDARs 383:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 9 theme GluN1 333:337 arg1 domains 365:371 GluN2B ligand binding domains 343:371 GluN2B ligand binding domains (LBDs) of NMDARs 343:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 9 theme GluN1 333:337 arg1 GluN1 333:337 GluN1 333:337 GluN1 333:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 4 10 theme intra-domain 449:460 arg1 interactions 462:473 intra-domain interactions 449:473 intra-domain interactions involving the glycan attached to residue GluN1-N440 449:525 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 5 11 theme similar 620:626 arg1 effect 644:649 a similar, though weaker, effect 618:649 effect 644:649 The glycan on GluN2B-N688 shows a similar, though weaker, effect. 28378791 1 12 theme N-methyl-D-aspartate 75:94 arg1 NMDARs 107:112 NMDARs 107:112 NMDARs 107:112 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 1 12 theme N-methyl-D-aspartate 75:94 arg1 glycoproteins 119:131 glycoproteins 119:131 glycoproteins in the brain central to learning and memory 119:175 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 1 12 theme N-methyl-D-aspartate 75:94 arg1 receptors 96:104 N-methyl-D-aspartate receptors 75:104 N-methyl-D-aspartate receptors (NMDARs) 75:113 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 8 13 from role 1105:1108 arg1 receptors 1129:1137 NMDA receptors 1124:1137 NMDA receptors 1124:1137 Overall, our results suggest an intramolecular potentiating role of glycans on NMDA receptors. 28378791 3 14 theme NMDARs 383:388 arg1 simulations 318:328 extensive molecular dynamics simulations 289:328 extensive molecular dynamics simulations of GluN1 289:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 14 theme NMDARs 383:388 arg1 LBDs 374:377 LBDs 374:377 LBDs 374:377 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 14 theme NMDARs 383:388 arg1 NMDARs 383:388 NMDARs 383:388 NMDARs 383:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 14 theme NMDARs 383:388 arg1 domains 365:371 GluN2B ligand binding domains 343:371 GluN2B ligand binding domains (LBDs) of NMDARs 343:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 14 theme NMDARs 383:388 arg1 GluN1 333:337 GluN1 333:337 GluN1 333:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 7 15 theme full-length 917:927 arg1 NMDARs 929:934 full-length NMDARs 917:934 full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation 917:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 0 16 theme Discovered 16:25 arg1 Role 40:43 Computationally Discovered Potentiating Role 0:43 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 0:73 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 28378791 7 17 with NMDARs 929:934 arg1 mutation 980:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 a glycosylation-preventing GluN1-N440Q mutation 941:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 7 18 theme NMDARs 929:934 arg1 analysis 905:912 electrophysiological analysis 884:912 electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation 884:987 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 6 19 from GluN1-N440 792:801 arg1 glycans 781:787 glycans 781:787 glycans at GluN1-N440 781:801 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 28378791 1 20 from glycoproteins 119:131 arg1 brain 140:144 the brain 136:144 the brain central to learning and memory 136:175 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 3 21 theme binding 357:363 arg1 LBDs 374:377 LBDs 374:377 LBDs 374:377 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 21 theme binding 357:363 arg1 NMDARs 383:388 NMDARs 383:388 NMDARs 383:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 21 theme binding 357:363 arg1 domains 365:371 GluN2B ligand binding domains 343:371 GluN2B ligand binding domains (LBDs) of NMDARs 343:388 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 3 21 theme binding 357:363 arg1 GluN1 333:337 GluN1 333:337 GluN1 333:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 0 22 theme Potentiating 27:38 arg1 Role 40:43 Computationally Discovered Potentiating Role 0:43 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 0:73 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 28378791 4 23 theme LBD 581:583 arg1 conformations 554:566 closed-clamshell conformations 537:566 closed-clamshell conformations of the GluN1 LBD 537:583 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 1 24 gly glycoproteins 119:131 arg1 glycoproteins 119:131 glycoproteins 119:131 glycoproteins in the brain central to learning and memory 119:175 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 1 24 gly glycoproteins 119:131 arg1 receptors 96:104 N-methyl-D-aspartate receptors 75:104 N-methyl-D-aspartate receptors (NMDARs) 75:113 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 2 25 dep structure 214:222 arg1 the 210:212 the 210:212 the 210:212 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 0 26 theme Glycans 48:54 arg1 Role 40:43 Computationally Discovered Potentiating Role 0:43 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 0:73 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 28378791 4 27 theme GluN1 575:579 arg1 LBD 581:583 the GluN1 LBD 571:583 the GluN1 LBD 571:583 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 8 28 theme potentiating 1092:1103 arg1 role 1105:1108 an intramolecular potentiating role 1074:1108 an intramolecular potentiating role of glycans on NMDA receptors 1074:1137 Overall, our results suggest an intramolecular potentiating role of glycans on NMDA receptors. 28378791 7 29 from increase 1009:1016 arg1 value 1038:1042 the glycine EC50 value 1021:1042 the glycine EC50 value 1021:1042 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 8 30 theme intramolecular 1077:1090 arg1 role 1105:1108 an intramolecular potentiating role 1074:1108 an intramolecular potentiating role of glycans on NMDA receptors 1074:1137 Overall, our results suggest an intramolecular potentiating role of glycans on NMDA receptors. 28378791 6 31 theme full 750:753 arg1 receptor 755:762 the full receptor 746:762 the full receptor 746:762 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 28378791 0 32 theme NMDA 59:62 arg1 Receptors 64:72 NMDA Receptors 59:72 NMDA Receptors 59:72 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 28378791 5 33 from glycan 590:595 arg1 GluN2B-N688 600:610 GluN2B-N688 600:610 GluN2B-N688 600:610 The glycan on GluN2B-N688 shows a similar, though weaker, effect. 28378791 3 34 theme extensive 289:297 arg1 simulations 318:328 extensive molecular dynamics simulations 289:328 extensive molecular dynamics simulations of GluN1 289:337 In this work, we use extensive molecular dynamics simulations of GluN1 and GluN2B ligand binding domains (LBDs) of NMDARs to investigate these effects. 28378791 6 35 theme potentiator 816:826 arg1 role 828:831 a potentiator role 814:831 a potentiator role 814:831 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 28378791 4 36 attach attached 496:503 arg2 glycan 489:494 the glycan 485:494 the glycan attached to residue GluN1-N440 485:525 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 4 36 attach attached 496:503 arg1 residue 508:514 residue GluN1-N440 508:525 residue GluN1-N440 508:525 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 4 37 theme closed-clamshell 537:552 arg1 conformations 554:566 closed-clamshell conformations 537:566 closed-clamshell conformations of the GluN1 LBD 537:583 Our simulations predict that intra-domain interactions involving the glycan attached to residue GluN1-N440 stabilize closed-clamshell conformations of the GluN1 LBD. 28378791 7 38 theme glycine 1025:1031 arg1 value 1038:1042 the glycine EC50 value 1021:1042 the glycine EC50 value 1021:1042 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 6 39 theme simulations 731:741 arg1 results 716:722 the results 712:722 the results of LBD simulations 712:741 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 28378791 5 40 dep similar 620:626 arg1 weaker 636:641 weaker 636:641 weaker 636:641 The glycan on GluN2B-N688 shows a similar, though weaker, effect. 28378791 7 41 theme EC50 1033:1036 arg1 value 1038:1042 the glycine EC50 value 1021:1042 the glycine EC50 value 1021:1042 To validate this prediction, we perform electrophysiological analysis of full-length NMDARs with a glycosylation-preventing GluN1-N440Q mutation, and demonstrate an increase in the glycine EC50 value. 28378791 2 42 theme glycosylation 193:205 arg1 effects 182:188 The effects 178:188 The effects of glycosylation on the structure and dynamics of NMDARs 178:245 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 2 42 theme glycosylation 193:205 arg1 unknown 259:265 unknown 259:265 unknown 259:265 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 6 43 theme LBD 727:729 arg1 simulations 731:741 LBD simulations 727:741 LBD simulations 727:741 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 28378791 1 44 theme central 146:152 arg1 brain 140:144 the brain 136:144 the brain central to learning and memory 136:175 N-methyl-D-aspartate receptors (NMDARs) are glycoproteins in the brain central to learning and memory. 28378791 2 45 gly glycosylation 193:205 arg1 NMDARs 240:245 NMDARs 240:245 NMDARs 240:245 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 8 46 theme NMDA 1124:1127 arg1 receptors 1129:1137 NMDA receptors 1124:1137 NMDA receptors 1124:1137 Overall, our results suggest an intramolecular potentiating role of glycans on NMDA receptors. 28378791 0 47 from Role 40:43 arg1 Receptors 64:72 NMDA Receptors 59:72 NMDA Receptors 59:72 Computationally Discovered Potentiating Role of Glycans on NMDA Receptors. 28378791 2 48 theme NMDARs 240:245 arg1 dynamics 228:235 dynamics 228:235 dynamics 228:235 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 2 48 theme NMDARs 240:245 arg1 structure 214:222 structure 214:222 structure 214:222 The effects of glycosylation on the structure and dynamics of NMDARs are largely unknown. 28378791 6 49 theme results 716:722 arg1 transferability 693:707 the transferability 689:707 the transferability of the results of LBD simulations to the full receptor 689:762 Based on these results, and assuming the transferability of the results of LBD simulations to the full receptor, we predict that glycans at GluN1-N440 might play a potentiator role in NMDARs. 25902405 3 0 gly N-glycopeptides 526:540 arg2 N-glycopeptides 526:540 N-glycopeptides 526:540 N-glycopeptides in the typical tryptic digests 526:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 3 1 from N-glycopeptides 526:540 arg1 digests 565:571 the typical tryptic digests 545:571 the typical tryptic digests 545:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 2 2 gly glycosylated 328:339 arg1 Asn 341:343 glycosylated Asn 328:343 glycosylated Asn at N-terminus (PGANs) 328:365 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 5 3 theme such 920:923 arg1 modification 925:936 such modification 920:936 such modification 920:936 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 2 4 from N-terminus 348:357 arg1 Asn 341:343 glycosylated Asn 328:343 glycosylated Asn at N-terminus (PGANs) 328:365 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 3 5 theme tryptic 557:563 arg1 digests 565:571 the typical tryptic digests 545:571 the typical tryptic digests 545:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 4 6 theme N-terminal 683:692 arg1 deglycosylation 742:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 5 7 theme modification 925:936 arg1 functions 907:915 the new biological functions 888:915 the new biological functions of such modification 888:936 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 5 8 theme glycoproteomes 854:867 arg1 analysis 842:849 the deep coverage analysis 824:849 the deep coverage analysis of glycoproteomes 824:867 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 3 9 from % 521:521 arg1 digests 565:571 the typical tryptic digests 545:571 the typical tryptic digests 545:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 5 10 theme obtained 763:770 arg1 information 778:788 The obtained PGANs information 759:788 The obtained PGANs information 759:788 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 5 10 theme obtained 763:770 arg1 beneficial 793:802 beneficial 793:802 beneficial 793:802 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 2 11 theme N-glycoproteome 287:301 arg1 profiling 303:311 N-glycoproteome profiling 287:311 N-glycoproteome profiling 287:311 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 1 12 theme global 190:195 arg1 analysis 213:220 the global N-glycoproteome analysis 186:220 the global N-glycoproteome analysis 186:220 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 2 13 theme deglycosylation 445:459 arg1 capacity 461:468 the poor deglycosylation capacity 436:468 the poor deglycosylation capacity by enzymes 436:479 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 5 14 theme PGANs 772:776 arg1 information 778:788 The obtained PGANs information 759:788 The obtained PGANs information 759:788 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 5 14 theme PGANs 772:776 arg1 beneficial 793:802 beneficial 793:802 beneficial 793:802 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 1 15 theme N-glycoproteome 197:211 arg1 analysis 213:220 the global N-glycoproteome analysis 186:220 the global N-glycoproteome analysis 186:220 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 2 16 theme poor 440:443 arg1 capacity 461:468 the poor deglycosylation capacity 436:468 the poor deglycosylation capacity by enzymes 436:479 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 1 17 from roles 128:132 arg1 processes 175:183 various biological processes 156:183 various biological processes 156:183 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 5 18 theme new 892:894 arg1 functions 907:915 the new biological functions 888:915 the new biological functions of such modification 888:936 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 0 19 theme peptide 24:30 arg1 N-terminus 32:41 peptide N-terminus 24:41 peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation 24:104 Releasing N-glycan from peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation. 25902405 5 20 theme deep 828:831 arg1 analysis 842:849 the deep coverage analysis 824:849 the deep coverage analysis of glycoproteomes 824:867 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 2 21 theme glycosylated 328:339 arg1 Asn 341:343 glycosylated Asn 328:343 glycosylated Asn at N-terminus (PGANs) 328:365 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 4 22 theme novel 615:619 arg1 strategy 621:628 a novel strategy 613:628 a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation 613:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 1 23 theme important 118:126 arg1 roles 128:132 the important roles 114:132 the important roles of N-glycoproteins in various biological processes 114:183 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 1 24 theme various 156:162 arg1 processes 175:183 various biological processes 156:183 various biological processes 156:183 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 1 25 gly N-glycoproteins 137:151 arg1 N-glycoproteins 137:151 N-glycoproteins 137:151 N-glycoproteins 137:151 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 5 26 theme coverage 833:840 arg1 analysis 842:849 the deep coverage analysis 824:849 the deep coverage analysis of glycoproteomes 824:867 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 1 27 theme much 236:239 arg1 attention 241:249 much attention 236:249 much attention 236:249 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 0 28 theme succinylation 57:69 arg1 deglycosylation 90:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 Releasing N-glycan from peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation. 25902405 5 29 theme biological 896:905 arg1 functions 907:915 the new biological functions 888:915 the new biological functions of such modification 888:936 The obtained PGANs information is beneficial to not only achieve the deep coverage analysis of glycoproteomes, but also discover the new biological functions of such modification. 25902405 4 30 theme succinylation 709:721 arg1 deglycosylation 742:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 0 31 theme N-terminal 46:55 arg1 deglycosylation 90:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 Releasing N-glycan from peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation. 25902405 4 32 theme site-selective 694:707 arg1 deglycosylation 742:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 1 33 theme N-glycoproteins 137:151 arg1 roles 128:132 the important roles 114:132 the important roles of N-glycoproteins in various biological processes 114:183 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 25902405 3 34 theme typical 549:555 arg1 digests 565:571 the typical tryptic digests 545:571 the typical tryptic digests 545:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 3 35 theme N-glycopeptides 526:540 arg1 N-glycopeptides 526:540 N-glycopeptides 526:540 N-glycopeptides in the typical tryptic digests 526:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 3 35 theme N-glycopeptides 526:540 arg1 % 521:521 10% 519:521 10% of N-glycopeptides in the typical tryptic digests 519:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 4 36 theme assisted 723:730 arg1 deglycosylation 742:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 3 37 from digests 565:571 arg1 N-glycopeptides 526:540 N-glycopeptides 526:540 N-glycopeptides in the typical tryptic digests 526:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 3 37 from digests 565:571 arg1 % 521:521 10% 519:521 10% of N-glycopeptides in the typical tryptic digests 519:571 However, theoretically, PGANs occupy 10% of N-glycopeptides in the typical tryptic digests. 25902405 4 38 theme enzymatic 732:740 arg1 deglycosylation 742:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 the N-terminal site-selective succinylation assisted enzymatic deglycosylation 679:756 Therefore, in this study, we developed a novel strategy to identify PGANs by releasing N-glycans through the N-terminal site-selective succinylation assisted enzymatic deglycosylation. 25902405 0 39 theme enzymatic 80:88 arg1 deglycosylation 90:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 Releasing N-glycan from peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation. 25902405 2 40 theme current 264:270 arg1 strategies 272:281 current strategies 264:281 current strategies for N-glycoproteome profiling 264:311 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 2 41 theme protease 381:388 arg1 digestion 390:398 protease digestion 381:398 protease digestion 381:398 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 2 42 with peptides 314:321 arg1 Asn 341:343 glycosylated Asn 328:343 glycosylated Asn at N-terminus (PGANs) 328:365 However, by current strategies for N-glycoproteome profiling, peptides with glycosylated Asn at N-terminus (PGANs), generated by protease digestion, could hardly be identified, due to the poor deglycosylation capacity by enzymes. 25902405 0 43 theme assisted 71:78 arg1 deglycosylation 90:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 N-terminal succinylation assisted enzymatic deglycosylation 46:104 Releasing N-glycan from peptide N-terminus by N-terminal succinylation assisted enzymatic deglycosylation. 25902405 1 44 theme biological 164:173 arg1 processes 175:183 various biological processes 156:183 various biological processes 156:183 Due to the important roles of N-glycoproteins in various biological processes, the global N-glycoproteome analysis has been paid much attention. 28437635 0 0 theme endogenous 63:72 arg1 PERV-A 88:93 PERV-A 88:93 PERV-A 88:93 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 0 0 theme endogenous 63:72 arg1 retrovirus-A 74:85 the porcine endogenous retrovirus-A 51:85 the porcine endogenous retrovirus-A (PERV-A) 51:94 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 6 1 theme either 824:829 arg1 Mutation 783:790 Mutation 783:790 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 783:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 3 2 theme cDNA 460:463 arg1 SLC52A1 474:480 the cDNA encoding SLC52A1 456:480 the cDNA encoding SLC52A1 456:480 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 0 3 theme porcine 55:61 arg1 PERV-A 88:93 PERV-A 88:93 PERV-A 88:93 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 0 3 theme porcine 55:61 arg1 retrovirus-A 74:85 the porcine endogenous retrovirus-A 51:85 the porcine endogenous retrovirus-A (PERV-A) 51:94 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 1 4 theme cross-species 288:300 arg1 transmission 302:313 cross-species transmission 288:313 cross-species transmission 288:313 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 3 5 theme encoding 465:472 arg1 SLC52A1 474:480 the cDNA encoding SLC52A1 456:480 the cDNA encoding SLC52A1 456:480 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 6 6 theme receptor 1045:1052 arg1 function 1054:1061 receptor function 1045:1061 receptor function 1045:1061 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 1 7 theme transmission 302:313 arg1 risk 280:283 the potential risk 266:283 the potential risk of cross-species transmission in xenotransplantation 266:336 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 6 8 theme C-terminal 867:876 arg1 tail 878:881 the C-terminal tail 863:881 the C-terminal tail at positions 442 or 446 863:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 4 9 theme SLC52A1 614:620 arg1 glycosylation 597:609 glycosylation 597:609 glycosylation of SLC52A1 597:620 In addition, we showed that glycosylation of SLC52A1 is not necessary for PERV-A receptor function. 28437635 6 10 theme surface 923:929 arg1 expression 931:940 SLC52A1 surface expression 915:940 SLC52A1 surface expression 915:940 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 4 11 theme receptor 650:657 arg1 function 659:666 PERV-A receptor function 643:666 PERV-A receptor function 643:666 In addition, we showed that glycosylation of SLC52A1 is not necessary for PERV-A receptor function. 28437635 1 12 theme human 231:235 arg1 cells 237:241 human cells 231:241 human cells 231:241 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 4 13 theme PERV-A 643:648 arg1 function 659:666 PERV-A receptor function 643:666 PERV-A receptor function 643:666 In addition, we showed that glycosylation of SLC52A1 is not necessary for PERV-A receptor function. 28437635 2 14 theme solute 358:363 arg1 family 373:378 the solute carrier family 52 354:381 the solute carrier family 52 (SLC52A1 and SLC52A2) 354:403 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 6 15 theme SLC52A1 915:921 arg1 expression 931:940 SLC52A1 surface expression 915:940 SLC52A1 surface expression 915:940 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 7 16 theme cellular 1114:1121 arg1 receptor 1123:1130 its cellular receptor 1110:1130 its cellular receptor 1110:1130 Understanding interactions between PERV-A and its cellular receptor may provide novel strategies to prevent zoonotic infection in the setting of xenotransplantation. 28437635 6 17 theme structural 1020:1029 arg1 stability 1031:1039 overall structural stability 1012:1039 overall structural stability 1012:1039 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 0 18 theme cysteine 6:13 arg1 residues 15:22 Three cysteine residues 0:22 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), 0:95 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 0 18 theme cysteine 6:13 arg1 SLC52A1 27:33 SLC52A1 27:33 SLC52A1 27:33 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 5 19 theme cysteine 713:720 arg1 residues 722:729 three cysteine residues 707:729 three cysteine residues 707:729 We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression. 28437635 3 20 theme glycosylation 523:535 arg1 signals 537:543 two putative glycosylation signals 510:543 two putative glycosylation signals 510:543 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 7 21 theme novel 1144:1148 arg1 strategies 1150:1159 novel strategies 1144:1159 novel strategies 1144:1159 Understanding interactions between PERV-A and its cellular receptor may provide novel strategies to prevent zoonotic infection in the setting of xenotransplantation. 28437635 6 22 theme overall 1012:1018 arg1 stability 1031:1039 overall structural stability 1012:1039 overall structural stability 1012:1039 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 5 23 theme cell 758:761 arg1 expression 771:780 SLC52A1 cell surface expression 750:780 SLC52A1 cell surface expression 750:780 We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression. 28437635 4 24 gly glycosylation 597:609 arg1 SLC52A1 614:620 SLC52A1 614:620 SLC52A1 614:620 In addition, we showed that glycosylation of SLC52A1 is not necessary for PERV-A receptor function. 28437635 3 25 theme signals 537:543 arg1 one 503:505 one 503:505 one 503:505 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 3 25 theme signals 537:543 arg1 signals 537:543 two putative glycosylation signals 510:543 two putative glycosylation signals 510:543 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 5 26 theme surface 763:769 arg1 expression 771:780 SLC52A1 cell surface expression 750:780 SLC52A1 cell surface expression 750:780 We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression. 28437635 6 27 from position 807:814 arg1 cysteine 795:802 cysteine 795:802 cysteine at position 365 795:818 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 27 from position 807:814 arg1 Mutation 783:790 Mutation 783:790 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 783:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 1 28 theme Porcine 162:168 arg1 PERV-A 195:200 PERV-A 195:200 PERV-A 195:200 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 1 28 theme Porcine 162:168 arg1 retrovirus-A 181:192 Porcine endogenous retrovirus-A 162:192 Porcine endogenous retrovirus-A (PERV-A) 162:201 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 1 28 theme Porcine 162:168 arg1 gammaretrovirus 206:220 a gammaretrovirus 204:220 a gammaretrovirus 204:220 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 3 29 theme putative 514:521 arg1 signals 537:543 two putative glycosylation signals 510:543 two putative glycosylation signals 510:543 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 1 30 theme endogenous 170:179 arg1 PERV-A 195:200 PERV-A 195:200 PERV-A 195:200 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 1 30 theme endogenous 170:179 arg1 retrovirus-A 181:192 Porcine endogenous retrovirus-A 162:192 Porcine endogenous retrovirus-A (PERV-A) 162:201 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 1 30 theme endogenous 170:179 arg1 gammaretrovirus 206:220 a gammaretrovirus 204:220 a gammaretrovirus 204:220 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 6 31 from Mutation 783:790 arg1 position 807:814 position 365 807:818 position 365 807:818 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 0 32 theme critical 104:111 arg1 role 113:116 a critical role 102:116 a critical role 102:116 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 2 33 dep family 373:378 arg1 SLC52A2 396:402 SLC52A2 396:402 SLC52A2 396:402 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 2 33 dep family 373:378 arg1 SLC52A1 384:390 SLC52A1 384:390 SLC52A1 384:390 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 2 34 theme family 373:378 arg1 members 343:349 Two members 339:349 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) 339:403 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 2 34 theme family 373:378 arg1 receptors 416:424 PERV-A receptors 409:424 PERV-A receptors 409:424 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 0 35 theme SLC52A1 27:33 arg1 residues 15:22 Three cysteine residues 0:22 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), 0:95 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 0 35 theme SLC52A1 27:33 arg1 SLC52A1 27:33 SLC52A1 27:33 SLC52A1 27:33 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 6 36 from 446 903:905 arg1 residues 851:858 the two cysteine residues 834:858 the two cysteine residues in the C-terminal tail at positions 442 or 446 834:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 36 from 446 903:905 arg1 tail 878:881 the C-terminal tail 863:881 the C-terminal tail at positions 442 or 446 863:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 2 37 theme carrier 365:371 arg1 family 373:378 the solute carrier family 52 354:381 the solute carrier family 52 (SLC52A1 and SLC52A2) 354:403 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 1 38 from risk 280:283 arg1 xenotransplantation 318:336 xenotransplantation 318:336 xenotransplantation 318:336 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 7 39 theme zoonotic 1172:1179 arg1 infection 1181:1189 zoonotic infection 1172:1189 zoonotic infection 1172:1189 Understanding interactions between PERV-A and its cellular receptor may provide novel strategies to prevent zoonotic infection in the setting of xenotransplantation. 28437635 6 40 from positions 886:894 arg1 residues 851:858 the two cysteine residues 834:858 the two cysteine residues in the C-terminal tail at positions 442 or 446 834:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 40 from positions 886:894 arg1 tail 878:881 the C-terminal tail 863:881 the C-terminal tail at positions 442 or 446 863:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 3 41 used occupied 548:555 arg2 signals 537:543 two putative glycosylation signals 510:543 two putative glycosylation signals 510:543 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 3 41 used occupied 548:555 arg2 one 503:505 one 503:505 one 503:505 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 6 42 theme cysteine 842:849 arg1 residues 851:858 the two cysteine residues 834:858 the two cysteine residues in the C-terminal tail at positions 442 or 446 834:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 43 theme cysteine 795:802 arg1 Mutation 783:790 Mutation 783:790 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 783:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 1 44 theme potential 270:278 arg1 risk 280:283 the potential risk 266:283 the potential risk of cross-species transmission in xenotransplantation 266:336 Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. 28437635 0 45 theme surface 126:132 arg1 expression 134:143 cell surface expression 121:143 cell surface expression 121:143 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 3 46 theme Site-directed 427:439 arg1 mutagenesis 441:451 Site-directed mutagenesis 427:451 Site-directed mutagenesis of the cDNA encoding SLC52A1 427:480 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 0 47 theme cell 121:124 arg1 expression 134:143 cell surface expression 121:143 cell surface expression 121:143 Three cysteine residues of SLC52A1, a receptor for the porcine endogenous retrovirus-A (PERV-A), play a critical role in cell surface expression and infectivity. 28437635 5 48 from minimum 698:704 arg1 sufficient 735:744 sufficient 735:744 sufficient 735:744 We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression. 28437635 6 49 from residues 851:858 arg1 positions 886:894 positions 442 or 446 886:905 positions 886:894 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 49 from residues 851:858 arg1 446 903:905 446 903:905 446 903:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 6 49 from residues 851:858 arg1 tail 878:881 the C-terminal tail 863:881 the C-terminal tail at positions 442 or 446 863:905 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 28437635 3 50 theme SLC52A1 474:480 arg1 mutagenesis 441:451 Site-directed mutagenesis 427:451 Site-directed mutagenesis of the cDNA encoding SLC52A1 427:480 Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. 28437635 2 51 theme PERV-A 409:414 arg1 members 343:349 Two members 339:349 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) 339:403 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 2 51 theme PERV-A 409:414 arg1 receptors 416:424 PERV-A receptors 409:424 PERV-A receptors 409:424 Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. 28437635 5 52 theme SLC52A1 750:756 arg1 expression 771:780 SLC52A1 cell surface expression 750:780 SLC52A1 cell surface expression 750:780 We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression. 28437635 7 53 theme xenotransplantation 1209:1227 arg1 setting 1198:1204 the setting 1194:1204 the setting of xenotransplantation 1194:1227 Understanding interactions between PERV-A and its cellular receptor may provide novel strategies to prevent zoonotic infection in the setting of xenotransplantation. 28437635 6 54 theme PERV-A 946:951 arg1 infection 953:961 PERV-A infection 946:961 PERV-A infection 946:961 Mutation of cysteine at position 365 and either of the two cysteine residues in the C-terminal tail at positions 442 or 446 reduced SLC52A1 surface expression and PERV-A infection suggesting that these residues may contribute to overall structural stability and receptor function. 25554420 0 0 theme multivalent 86:96 arg1 binding 105:111 multivalent glycan binding 86:111 multivalent glycan binding 86:111 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 3 1 theme cytotoxic 714:722 arg1 activity 724:731 cytotoxic activity 714:731 cytotoxic activity of CyaA 714:739 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 2 theme located 634:640 arg1 N-glycans 624:632 N-glycans 624:632 N-glycans located in the C-terminal part of the CD11b subunit 624:684 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 1 3 theme toxin 172:176 arg1 interaction 118:128 The interaction 114:128 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) 114:228 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 1 4 theme N-linked 239:246 arg1 chains 264:269 N-linked oligosaccharide chains 239:269 N-linked oligosaccharide chains 239:269 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 2 5 theme toxin 350:354 arg1 activity 356:363 toxin activity 350:363 toxin activity 350:363 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 1 6 theme oligosaccharide 248:262 arg1 chains 264:269 N-linked oligosaccharide chains 239:269 N-linked oligosaccharide chains 239:269 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 4 7 theme clustered 789:797 arg1 patch 810:814 a defined clustered saccharide patch 779:814 a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction 779:939 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 3 8 theme mutant 525:530 arg1 variants 532:539 CR3 mutant variants 521:539 CR3 mutant variants 521:539 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 2 9 theme relative 291:298 arg1 importance 300:309 the relative importance 287:309 the relative importance of the individual N-glycans of CR3 for toxin activity 287:363 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 0 10 theme glycan 98:103 arg1 binding 105:111 multivalent glycan binding 86:111 multivalent glycan binding 86:111 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 3 11 theme CyaA 736:739 arg1 binding 702:708 binding 702:708 binding 702:708 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 11 theme CyaA 736:739 arg1 activity 724:731 cytotoxic activity 714:731 cytotoxic activity of CyaA 714:739 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 2 12 gly N-glycosylation 407:421 arg2 CR3 432:434 CR3 432:434 CR3 432:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 12 gly N-glycosylation 407:421 arg1 CR3 432:434 CR3 432:434 CR3 432:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 12 gly N-glycosylation 407:421 arg2 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 1 13 link N-linked 239:246 arg1 chains 264:269 N-linked oligosaccharide chains 239:269 N-linked oligosaccharide chains 239:269 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 2 14 theme N-glycans 329:337 arg1 importance 300:309 the relative importance 287:309 the relative importance of the individual N-glycans of CR3 for toxin activity 287:363 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 4 15 theme defined 781:787 arg1 patch 810:814 a defined clustered saccharide patch 779:814 a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction 779:939 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 3 16 from located 634:640 arg1 part 660:663 the C-terminal part 645:663 the C-terminal part of the CD11b subunit 645:684 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 4 17 theme CR3 852:854 arg1 contact 841:847 multivalent contact 829:847 multivalent contact of CR3 with CyaA 829:864 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 3 18 theme mass 545:548 arg1 analysis 563:570 mass spectrometry analysis 545:570 mass spectrometry analysis of the N-glycosylation pattern of CR3 545:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 19 theme variants 532:539 arg1 analysis 563:570 mass spectrometry analysis 545:570 mass spectrometry analysis of the N-glycosylation pattern of CR3 545:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 19 theme variants 532:539 arg1 Examination 506:516 Examination 506:516 Examination of CR3 mutant variants 506:539 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 1 20 with interaction 118:128 arg1 receptor 201:208 complement receptor 3 190:210 complement receptor 3 (CR3, CD11b/CD18) 190:228 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 2 21 theme individual 318:327 arg1 N-glycans 329:337 the individual N-glycans 314:337 the individual N-glycans of CR3 314:344 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 3 22 theme pattern 595:601 arg1 analysis 563:570 mass spectrometry analysis 545:570 mass spectrometry analysis of the N-glycosylation pattern of CR3 545:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 22 theme pattern 595:601 arg1 Examination 506:516 Examination 506:516 Examination of CR3 mutant variants 506:539 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 1 23 theme complement 190:199 arg1 receptor 201:208 complement receptor 3 190:210 complement receptor 3 (CR3, CD11b/CD18) 190:228 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 0 24 theme adenylate 26:34 arg1 cyclase 36:42 Bordetella adenylate cyclase 15:42 Bordetella adenylate cyclase toxin 15:48 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 0 25 theme Bordetella 15:24 arg1 cyclase 36:42 Bordetella adenylate cyclase 15:42 Bordetella adenylate cyclase toxin 15:48 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 4 26 theme interaction 929:939 arg1 specificity 895:905 specificity 895:905 specificity of the integrin-toxin interaction 895:939 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 4 26 theme interaction 929:939 arg1 affinity 882:889 affinity 882:889 affinity 882:889 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 2 27 gly glycosylated 492:503 arg1 residues 468:475 glutamine residues 458:475 glutamine residues that cannot be glycosylated 458:503 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 28 theme sites 423:427 arg1 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 28 theme sites 423:427 arg1 residues 381:388 the asparagine residues 366:388 the asparagine residues of the consensus N-glycosylation sites of CR3 366:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 0 29 theme toxin 44:48 arg1 Interaction 0:10 Interaction 0:10 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 0:75 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 3 30 gly N-glycosylation 579:593 arg1 CR3 606:608 CR3 606:608 CR3 606:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 4 31 theme multivalent 829:839 arg1 contact 841:847 multivalent contact 829:847 multivalent contact of CR3 with CyaA 829:864 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 2 32 theme asparagine 370:379 arg1 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 32 theme asparagine 370:379 arg1 residues 381:388 the asparagine residues 366:388 the asparagine residues of the consensus N-glycosylation sites of CR3 366:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 33 theme N-glycosylation 407:421 arg1 CR3 432:434 CR3 432:434 CR3 432:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 33 theme N-glycosylation 407:421 arg1 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 0 34 theme cyclase 36:42 arg1 toxin 44:48 Bordetella adenylate cyclase toxin 15:48 Bordetella adenylate cyclase toxin 15:48 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 3 35 theme C-terminal 649:658 arg1 part 660:663 the C-terminal part 645:663 the C-terminal part of the CD11b subunit 645:684 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 2 36 theme consensus 397:405 arg1 CR3 432:434 CR3 432:434 CR3 432:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 36 theme consensus 397:405 arg1 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 0 37 theme complement 55:64 arg1 receptor 66:73 complement receptor 3 55:75 complement receptor 3 55:75 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 2 38 theme glutamine 458:466 arg1 residues 468:475 glutamine residues 458:475 glutamine residues that cannot be glycosylated 458:503 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 3 39 theme CR3 606:608 arg1 pattern 595:601 the N-glycosylation pattern 575:601 the N-glycosylation pattern of CR3 575:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 40 theme CR3 521:523 arg1 variants 532:539 CR3 mutant variants 521:539 CR3 mutant variants 521:539 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 2 41 theme CR3 342:344 arg1 N-glycans 329:337 the individual N-glycans 314:337 the individual N-glycans of CR3 314:344 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 3 42 theme N-glycosylation 579:593 arg1 pattern 595:601 the N-glycosylation pattern 575:601 the N-glycosylation pattern of CR3 575:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 4 43 theme integrin-toxin 914:927 arg1 interaction 929:939 the integrin-toxin interaction 910:939 the integrin-toxin interaction 910:939 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 4 44 theme saccharide 799:808 arg1 patch 810:814 a defined clustered saccharide patch 779:814 a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction 779:939 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 0 45 with Interaction 0:10 arg1 receptor 66:73 complement receptor 3 55:75 complement receptor 3 55:75 Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding. 25554420 1 46 theme Bordetella 133:142 arg1 cyclase 164:170 Bordetella pertussis adenylate cyclase 133:170 Bordetella pertussis adenylate cyclase toxin (CyaA) 133:183 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 3 47 theme CD11b 672:676 arg1 subunit 678:684 the CD11b subunit 668:684 the CD11b subunit 668:684 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 3 48 theme spectrometry 550:561 arg1 analysis 563:570 mass spectrometry analysis 545:570 mass spectrometry analysis of the N-glycosylation pattern of CR3 545:608 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 1 49 theme pertussis 144:152 arg1 cyclase 164:170 Bordetella pertussis adenylate cyclase 133:170 Bordetella pertussis adenylate cyclase toxin (CyaA) 133:183 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 4 50 with contact 841:847 arg1 CyaA 861:864 CyaA 861:864 CyaA 861:864 We suggest that these N-glycans form a defined clustered saccharide patch that enables multivalent contact of CR3 with CyaA, enhancing both affinity and specificity of the integrin-toxin interaction. 25554420 3 51 theme subunit 678:684 arg1 part 660:663 the C-terminal part 645:663 the C-terminal part of the CD11b subunit 645:684 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 25554420 1 52 dep receptor 201:208 arg1 CD11b/CD18 218:227 CD11b/CD18 218:227 CD11b/CD18 218:227 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 1 52 dep receptor 201:208 arg1 CR3 213:215 CR3 213:215 CR3 213:215 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 1 53 theme adenylate 154:162 arg1 cyclase 164:170 Bordetella pertussis adenylate cyclase 133:170 Bordetella pertussis adenylate cyclase toxin (CyaA) 133:183 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 2 54 theme CR3 432:434 arg1 CR3 432:434 CR3 432:434 CR3 432:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 2 54 theme CR3 432:434 arg1 sites 423:427 the consensus N-glycosylation sites 393:427 the consensus N-glycosylation sites of CR3 393:434 To investigate the relative importance of the individual N-glycans of CR3 for toxin activity, the asparagine residues of the consensus N-glycosylation sites of CR3 were substituted with glutamine residues that cannot be glycosylated. 25554420 1 55 theme cyclase 164:170 arg1 CyaA 179:182 CyaA 179:182 CyaA 179:182 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 1 55 theme cyclase 164:170 arg1 toxin 172:176 Bordetella pertussis adenylate cyclase toxin 133:176 Bordetella pertussis adenylate cyclase toxin (CyaA) 133:183 The interaction of Bordetella pertussis adenylate cyclase toxin (CyaA) with complement receptor 3 (CR3, CD11b/CD18) involves N-linked oligosaccharide chains. 25554420 3 56 from part 660:663 arg1 located 634:640 located 634:640 located 634:640 Examination of CR3 mutant variants and mass spectrometry analysis of the N-glycosylation pattern of CR3 revealed that N-glycans located in the C-terminal part of the CD11b subunit are involved in binding and cytotoxic activity of CyaA. 26105115 5 0 theme mannose 766:772 arg1 patch 774:778 the intrinsic mannose patch 752:778 the intrinsic mannose patch 752:778 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 5 1 from changes 793:799 arg1 processing 808:817 the processing 804:817 the processing of neighbouring glycans 804:841 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 5 2 theme patch 774:778 arg1 size 744:747 the overall size 732:747 the overall size of the intrinsic mannose patch 732:778 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 6 3 theme glycan-dependent 905:920 arg1 bnAbs 922:926 glycan-dependent bnAbs 905:926 glycan-dependent bnAbs targeting these regions 905:950 These structural changes are largely tolerated by a panel of glycan-dependent bnAbs targeting these regions, indicating a degree of plasticity in their recognition. 26105115 6 4 theme structural 850:859 arg1 changes 861:867 These structural changes 844:867 These structural changes 844:867 These structural changes are largely tolerated by a panel of glycan-dependent bnAbs targeting these regions, indicating a degree of plasticity in their recognition. 26105115 0 5 theme neutralizing 95:106 arg1 antibodies 108:117 broadly neutralizing antibodies 87:117 broadly neutralizing antibodies 87:117 Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. 26105115 3 6 theme bnAbs 541:545 arg1 region 450:455 a region 448:455 a region 448:455 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 3 6 theme bnAbs 541:545 arg1 target 525:530 a primary target 515:530 a primary target of these bnAbs 515:545 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 4 7 theme glycosylation 599:611 arg1 sites 613:617 individual glycosylation sites 588:617 individual glycosylation sites 588:617 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 3 8 theme high 360:363 arg1 density 372:378 The unusually high glycan density 346:378 The unusually high glycan density on the gp120 subunit 346:399 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 4 9 theme individual 588:597 arg1 sites 613:617 individual glycosylation sites 588:617 individual glycosylation sites 588:617 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 3 10 from density 372:378 arg1 subunit 393:399 the gp120 subunit 383:399 the gp120 subunit 383:399 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 5 11 theme glycans 835:841 arg1 processing 808:817 the processing 804:817 the processing of neighbouring glycans 804:841 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 4 12 theme patch 672:676 arg1 formation 626:634 the formation 622:634 the formation of this so-called intrinsic mannose patch 622:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 4 13 from contribution 572:583 arg1 formation 626:634 the formation 622:634 the formation of this so-called intrinsic mannose patch 622:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 4 14 theme mannose 664:670 arg1 patch 672:676 this so-called intrinsic mannose patch 639:676 this so-called intrinsic mannose patch 639:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 7 15 theme vaccine 1082:1088 arg1 design 1090:1095 vaccine design 1082:1095 vaccine design 1082:1095 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 0 16 theme Glycan 0:5 arg1 clustering 7:16 Glycan clustering 0:16 Glycan clustering 0:16 Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. 26105115 6 17 theme plasticity 976:985 arg1 degree 966:971 a degree 964:971 a degree of plasticity in their recognition 964:1006 These structural changes are largely tolerated by a panel of glycan-dependent bnAbs targeting these regions, indicating a degree of plasticity in their recognition. 26105115 2 18 dep potent 255:260 arg1 neutralizing 270:281 neutralizing 270:281 neutralizing 270:281 Paradoxically, however, potent broadly neutralizing antibodies (bnAbs) that target this shield have been isolated. 26105115 3 19 theme glycan 365:370 arg1 density 372:378 The unusually high glycan density 346:378 The unusually high glycan density on the gp120 subunit 346:399 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 6 20 from degree 966:971 arg1 recognition 996:1006 their recognition 990:1006 their recognition 990:1006 These structural changes are largely tolerated by a panel of glycan-dependent bnAbs targeting these regions, indicating a degree of plasticity in their recognition. 26105115 4 21 gly glycosylation 599:611 arg2 sites 613:617 individual glycosylation sites 588:617 individual glycosylation sites 588:617 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 5 22 theme neighbouring 822:833 arg1 glycans 835:841 neighbouring glycans 822:841 neighbouring glycans 822:841 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 5 23 theme limited 714:720 arg1 effect 722:727 a limited effect 712:727 a limited effect 712:727 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 6 24 theme bnAbs 922:926 arg1 panel 896:900 a panel 894:900 a panel of glycan-dependent bnAbs targeting these regions 894:950 These structural changes are largely tolerated by a panel of glycan-dependent bnAbs targeting these regions, indicating a degree of plasticity in their recognition. 26105115 3 25 theme under-processed 460:474 arg1 structures 494:503 under-processed oligomannose-type structures 460:503 under-processed oligomannose-type structures 460:503 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 4 26 theme intrinsic 654:662 arg1 patch 672:676 this so-called intrinsic mannose patch 639:676 this so-called intrinsic mannose patch 639:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 0 27 theme mannose 33:39 arg1 patch 41:45 the mannose patch 29:45 the mannose patch of HIV-1 29:54 Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. 26105115 5 28 contain has 708:710 arg2 effect 722:727 a limited effect 712:727 a limited effect 712:727 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 5 28 contain has 708:710 arg1 Deletion 679:686 Deletion 679:686 Deletion of individual sites 679:706 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 7 29 theme stable 1064:1069 arg1 patch 1053:1057 the intrinsic mannose patch 1031:1057 the intrinsic mannose patch 1031:1057 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 7 29 theme stable 1064:1069 arg1 target 1071:1076 a stable target 1062:1076 a stable target for vaccine design 1062:1095 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 1 30 theme antibody-mediated 197:213 arg1 neutralization 215:228 antibody-mediated neutralization 197:228 antibody-mediated neutralization 197:228 The envelope spike of HIV-1 employs a 'glycan shield' to protect itself from antibody-mediated neutralization. 26105115 0 31 theme HIV-1 50:54 arg1 patch 41:45 the mannose patch 29:45 the mannose patch of HIV-1 29:54 Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies. 26105115 5 32 theme intrinsic 756:764 arg1 patch 774:778 the intrinsic mannose patch 752:778 the intrinsic mannose patch 752:778 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 7 33 theme intrinsic 1035:1043 arg1 patch 1053:1057 the intrinsic mannose patch 1031:1057 the intrinsic mannose patch 1031:1057 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 7 33 theme intrinsic 1035:1043 arg1 target 1071:1076 a stable target 1062:1076 a stable target for vaccine design 1062:1095 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 5 34 theme individual 691:700 arg1 sites 702:706 individual sites 691:706 individual sites 691:706 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 1 35 theme envelope 124:131 arg1 spike 133:137 The envelope spike 120:137 The envelope spike of HIV-1 120:146 The envelope spike of HIV-1 employs a 'glycan shield' to protect itself from antibody-mediated neutralization. 26105115 5 36 theme sites 702:706 arg1 Deletion 679:686 Deletion 679:686 Deletion of individual sites 679:706 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 26105115 1 37 theme HIV-1 142:146 arg1 spike 133:137 The envelope spike 120:137 The envelope spike of HIV-1 120:146 The envelope spike of HIV-1 employs a 'glycan shield' to protect itself from antibody-mediated neutralization. 26105115 3 38 theme oligomannose-type 476:492 arg1 structures 494:503 under-processed oligomannose-type structures 460:503 under-processed oligomannose-type structures 460:503 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 7 39 theme mannose 1045:1051 arg1 patch 1053:1057 the intrinsic mannose patch 1031:1057 the intrinsic mannose patch 1031:1057 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 7 39 theme mannose 1045:1051 arg1 target 1071:1076 a stable target 1062:1076 a stable target for vaccine design 1062:1095 These results support the intrinsic mannose patch as a stable target for vaccine design. 26105115 2 40 theme potent 255:260 arg1 bnAbs 295:299 bnAbs 295:299 bnAbs 295:299 Paradoxically, however, potent broadly neutralizing antibodies (bnAbs) that target this shield have been isolated. 26105115 2 40 theme potent 255:260 arg1 antibodies 283:292 potent broadly neutralizing antibodies 255:292 potent broadly neutralizing antibodies (bnAbs) that target this shield 255:324 Paradoxically, however, potent broadly neutralizing antibodies (bnAbs) that target this shield have been isolated. 26105115 3 41 theme structures 494:503 arg1 region 450:455 a region 448:455 a region 448:455 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 3 41 theme structures 494:503 arg1 target 525:530 a primary target 515:530 a primary target of these bnAbs 515:545 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 4 42 theme sites 613:617 arg1 contribution 572:583 the contribution 568:583 the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch 568:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 4 43 theme so-called 644:652 arg1 patch 672:676 this so-called intrinsic mannose patch 639:676 this so-called intrinsic mannose patch 639:676 Here we investigate the contribution of individual glycosylation sites in the formation of this so-called intrinsic mannose patch. 26105115 3 44 theme primary 517:523 arg1 region 450:455 a region 448:455 a region 448:455 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 3 44 theme primary 517:523 arg1 target 525:530 a primary target 515:530 a primary target of these bnAbs 515:545 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 3 45 theme gp120 387:391 arg1 subunit 393:399 the gp120 subunit 383:399 the gp120 subunit 383:399 The unusually high glycan density on the gp120 subunit limits processing during biosynthesis, leaving a region of under-processed oligomannose-type structures, which is a primary target of these bnAbs. 26105115 5 46 theme overall 736:742 arg1 size 744:747 the overall size 732:747 the overall size of the intrinsic mannose patch 732:778 Deletion of individual sites has a limited effect on the overall size of the intrinsic mannose patch but leads to changes in the processing of neighbouring glycans. 27957769 11 0 theme immune-dominant 2219:2233 arg1 PDTR 2235:2238 the immune-dominant PDTR 2215:2238 the immune-dominant PDTR 2215:2238 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 6 1 theme libraries 1313:1321 arg1 one 1274:1276 one 1274:1276 one 1274:1276 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 6 1 theme libraries 1313:1321 arg1 libraries 1313:1321 the most extensive glycopeptide libraries 1281:1321 the most extensive glycopeptide libraries ever made through total synthesis 1281:1355 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 8 2 theme such 1757:1760 arg1 epitopes 1762:1769 such epitopes 1757:1769 such epitopes 1757:1769 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 5 3 from influence 789:797 arg1 recognition 835:845 antibody recognition 826:845 antibody recognition 826:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 4 dep structures 1094:1103 arg1 type-1 1106:1111 type-1 1106:1111 type-1 1106:1111 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 4 dep structures 1094:1103 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 4 dep structures 1094:1103 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 10 5 theme MUC1 2029:2032 arg1 peptides 2034:2041 MUC1 peptides 2029:2041 MUC1 peptides 2029:2041 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 6 6 theme extensive 1290:1298 arg1 libraries 1313:1321 the most extensive glycopeptide libraries 1281:1321 the most extensive glycopeptide libraries ever made through total synthesis 1281:1355 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 5 7 theme tandem 1251:1256 arg1 repeat 1258:1263 the MUC1 tandem repeat 1242:1263 the MUC1 tandem repeat 1242:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 11 8 theme GSTA 2247:2250 arg1 domain 2252:2257 the GSTA domain 2243:2257 the GSTA domain 2243:2257 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 7 9 theme lower 1471:1475 arg1 structures 1508:1517 the branched core 2 structures 1488:1517 the branched core 2 structures 1488:1517 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 7 9 theme lower 1471:1475 arg1 amounts 1477:1483 lower amounts 1471:1483 lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures 1471:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 4 10 theme immune 718:723 arg1 stimulants 725:734 different immune stimulants 708:734 different immune stimulants 708:734 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 3 11 with antibodies 529:538 arg1 cross-reactivity 545:560 cross-reactivity 545:560 cross-reactivity 545:560 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 1 12 theme biomarker 189:197 arg1 discovery 199:207 biomarker discovery 189:207 biomarker discovery 189:207 Glycoprotein research is pivotal for vaccine development and biomarker discovery. 27957769 7 13 theme core 2 1501:1506 arg1 structures 1508:1517 the branched core 2 structures 1488:1517 the branched core 2 structures 1488:1517 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 10 14 theme high 2053:2056 arg1 specificity 2077:2087 very high glycosylation site specificity 2048:2087 very high glycosylation site specificity 2048:2087 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 12 15 from structures 2334:2343 arg1 MUC1 2348:2351 MUC1 2348:2351 MUC1 2348:2351 All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. 27957769 5 16 theme type-2 1117:1122 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 16 theme type-2 1117:1122 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 3 17 theme autoimmune 610:619 arg1 diseases 621:628 autoimmune diseases 610:628 autoimmune diseases 610:628 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 13 18 theme healthy 2430:2436 arg1 cells 2438:2442 healthy cells 2430:2442 healthy cells 2430:2442 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 5 19 theme influence 789:797 arg1 assessment 771:780 assessment 771:780 assessment of the influence of the immune stimulant on antibody recognition 771:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 20 theme core 1134:1137 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 20 theme core 1134:1137 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 21 theme glycopeptides 883:895 arg1 library 864:870 a comprehensive library 848:870 a comprehensive library of mucin 1 glycopeptides (>100 entries) 848:910 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 8 22 theme vaccines 1829:1836 arg1 safety 1809:1814 safety 1809:1814 safety 1809:1814 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 8 22 theme vaccines 1829:1836 arg1 tumor-selectivity 1787:1803 tumor-selectivity 1787:1803 tumor-selectivity 1787:1803 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 10 23 theme site 2072:2075 arg1 specificity 2077:2087 very high glycosylation site specificity 2048:2087 very high glycosylation site specificity 2048:2087 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 6 24 theme total 1341:1345 arg1 synthesis 1347:1355 total synthesis 1341:1355 total synthesis 1341:1355 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 5 25 from recognition 835:845 arg1 assessment 771:780 assessment 771:780 assessment of the influence of the immune stimulant on antibody recognition 771:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 1 26 theme Glycoprotein 128:139 arg1 research 141:148 Glycoprotein research 128:148 Glycoprotein research 128:148 Glycoprotein research is pivotal for vaccine development and biomarker discovery. 27957769 3 27 theme Deeper 349:354 arg1 insights 356:363 Deeper insights 349:363 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions 349:440 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 7 28 theme core 2 1378:1383 arg1 C2GlcNAcT-1 1426:1436 C2GlcNAcT-1 1426:1436 C2GlcNAcT-1 1426:1436 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 7 28 theme core 2 1378:1383 arg1 β-1,6-N-acetylglucosaminyltransferase-1 1385:1423 the core 2 β-1,6-N-acetylglucosaminyltransferase-1 1374:1423 the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) 1374:1437 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 2 29 theme glycopeptide 305:316 arg1 structures 318:327 tumor-associated glycopeptide structures 288:327 tumor-associated glycopeptide structures 288:327 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. 27957769 5 30 theme small 991:995 arg1 glycans 1014:1020 small tumor-associated glycans 991:1020 small tumor-associated glycans (TN , STN , and T-antigen structures) 991:1058 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 11 31 gly glycosylation 2185:2197 arg2 sites 2199:2203 glycosylation sites 2185:2203 glycosylation sites in either the immune-dominant PDTR or the GSTA domain 2185:2257 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 11 32 from sites 2199:2203 arg1 PDTR 2235:2238 the immune-dominant PDTR 2215:2238 the immune-dominant PDTR 2215:2238 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 11 32 from sites 2199:2203 arg1 domain 2252:2257 the GSTA domain 2243:2257 the GSTA domain 2243:2257 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 5 33 theme MUC1 1246:1249 arg1 repeat 1258:1263 the MUC1 tandem repeat 1242:1263 the MUC1 tandem repeat 1242:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 34 theme microarray 952:961 arg1 profiling 963:971 antibody microarray profiling 943:971 antibody microarray profiling 943:971 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 6 35 gly glycopeptide 1300:1311 arg2 glycopeptide 1300:1311 the most extensive glycopeptide libraries 1281:1321 the most extensive glycopeptide libraries ever made through total synthesis 1281:1355 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 7 36 theme tumor-associated 1610:1625 arg1 structures 1635:1644 truncated tumor-associated antigen structures 1600:1644 truncated tumor-associated antigen structures 1600:1644 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 3 37 theme polyclonal 412:421 arg1 reactions 432:440 the raised polyclonal, humoral reactions 401:440 the raised polyclonal, humoral reactions 401:440 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 13 38 theme tumor-specific 2557:2570 arg1 response 2587:2594 a tumor-specific humoral immune response 2555:2594 a tumor-specific humoral immune response 2555:2594 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 0 39 theme Diverse 97:103 arg1 Structures 116:125 Diverse Mucin Core Structures 97:125 Diverse Mucin Core Structures 97:125 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 13 40 theme immune 2580:2585 arg1 response 2587:2594 a tumor-specific humoral immune response 2555:2594 a tumor-specific humoral immune response 2555:2594 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 9 41 theme mucin 1857:1861 arg1 structures 1868:1877 the extended mucin core structures 1844:1877 the extended mucin core structures in hand 1844:1885 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 8 42 dep tumor-selectivity 1787:1803 arg1 the 1783:1785 the 1783:1785 the 1783:1785 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 5 43 theme stimulant 813:821 arg1 influence 789:797 the influence 785:797 the influence of the immune stimulant on antibody recognition 785:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 12 44 theme tumor-associated 2306:2321 arg1 structures 2334:2343 the tumor-associated saccharide structures 2302:2343 the tumor-associated saccharide structures on MUC1 2302:2351 All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. 27957769 0 45 theme Microarray 0:9 arg1 Analysis 11:18 Microarray Analysis 0:18 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 0:126 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 3 46 dep quality 374:380 arg1 the 370:372 the 370:372 the 370:372 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 5 47 theme different 1223:1231 arg1 repeat 1258:1263 the MUC1 tandem repeat 1242:1263 the MUC1 tandem repeat 1242:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 47 theme different 1223:1231 arg1 sites 1233:1237 the five different sites 1214:1237 the five different sites of the MUC1 tandem repeat 1214:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 48 theme TN 1023:1024 arg1 STN 1028:1030 TN , STN , and T-antigen structures 1023:1057 STN 1028:1030 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 9 49 theme antibody 1888:1895 arg1 cross-reactivity 1897:1912 antibody cross-reactivity 1888:1912 antibody cross-reactivity toward the branched core 2 glycopeptide epitopes 1888:1961 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 12 50 theme high 2283:2286 arg1 reactivity 2288:2297 high reactivity 2283:2297 high reactivity to the tumor-associated saccharide structures on MUC1 2283:2351 All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. 27957769 3 51 theme immunological 492:504 arg1 memory 506:511 an immunological memory 489:511 an immunological memory 489:511 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 5 52 theme comprehensive 850:862 arg1 library 864:870 a comprehensive library 848:870 a comprehensive library of mucin 1 glycopeptides (>100 entries) 848:910 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 7 53 theme structures 1569:1578 arg1 formation 1532:1540 formation 1532:1540 formation of linear core 1 or core 3 structures 1532:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 5 54 theme extended 1071:1078 arg1 type-1 1106:1111 type-1 1106:1111 type-1 1106:1111 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 54 theme extended 1071:1078 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 54 theme extended 1071:1078 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 55 theme antibody 826:833 arg1 recognition 835:845 antibody recognition 826:845 antibody recognition 826:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 56 dep synthesized 915:925 arg1 range 980:984 these range 974:984 these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 974:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 9 57 from structures 1868:1877 arg1 hand 1882:1885 hand 1882:1885 hand 1882:1885 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 0 58 theme Synthetic 47:55 arg1 Vaccines 67:74 Synthetic Antitumor Vaccines 47:74 Synthetic Antitumor Vaccines 47:74 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 7 59 from structures 1635:1644 arg1 structures 1508:1517 the branched core 2 structures 1488:1517 the branched core 2 structures 1488:1517 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 7 59 from structures 1635:1644 arg1 amounts 1477:1483 lower amounts 1471:1483 lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures 1471:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 5 60 theme core 1089:1092 arg1 type-1 1106:1111 type-1 1106:1111 type-1 1106:1111 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 60 theme core 1089:1092 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 60 theme core 1089:1092 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 9 61 theme branched 1925:1932 arg1 epitopes 1954:1961 the branched core 2 glycopeptide epitopes 1921:1961 the branched core 2 glycopeptide epitopes 1921:1961 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 3 62 dep polyclonal 412:421 arg1 humoral 424:430 humoral 424:430 humoral 424:430 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 5 63 theme repeat 1258:1263 arg1 repeat 1258:1263 the MUC1 tandem repeat 1242:1263 the MUC1 tandem repeat 1242:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 63 theme repeat 1258:1263 arg1 sites 1233:1237 the five different sites 1214:1237 the five different sites of the MUC1 tandem repeat 1214:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 7 64 theme linear 1545:1550 arg1 core 1 1552:1557 linear core 1 1545:1557 linear core 1 1545:1557 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 9 65 theme core 1863:1866 arg1 structures 1868:1877 the extended mucin core structures 1844:1877 the extended mucin core structures in hand 1844:1885 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 9 66 theme glycopeptide 1941:1952 arg1 epitopes 1954:1961 the branched core 2 glycopeptide epitopes 1921:1961 the branched core 2 glycopeptide epitopes 1921:1961 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 8 67 theme core 2 1651:1656 arg1 structures 1658:1667 The core 2 structures 1647:1667 The core 2 structures 1647:1667 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 2 68 theme Many 210:213 arg1 methodologies 226:238 Many successful methodologies 210:238 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures 210:327 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. 27957769 5 69 from glycans 1014:1020 arg1 range 980:984 these range 974:984 these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 974:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 4 70 theme MUC1 658:661 arg1 candidates 681:690 three MUC1 antitumor vaccine candidates 652:690 three MUC1 antitumor vaccine candidates conjugated with different immune stimulants 652:734 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 8 71 theme antibody 1728:1735 arg1 cross-reactivity 1737:1752 antibody cross-reactivity 1728:1752 antibody cross-reactivity 1728:1752 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 6 72 theme glycopeptide 1300:1311 arg1 libraries 1313:1321 the most extensive glycopeptide libraries 1281:1321 the most extensive glycopeptide libraries ever made through total synthesis 1281:1355 This is one of the most extensive glycopeptide libraries ever made through total synthesis. 27957769 13 73 theme vaccine 2514:2520 arg1 conjugates 2522:2531 all vaccine conjugates 2510:2531 all vaccine conjugates 2510:2531 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 5 74 from assessment 771:780 arg1 recognition 835:845 antibody recognition 826:845 antibody recognition 826:845 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 7 75 theme structures 1508:1517 arg1 structures 1508:1517 the branched core 2 structures 1488:1517 the branched core 2 structures 1488:1517 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 7 75 theme structures 1508:1517 arg1 amounts 1477:1483 lower amounts 1471:1483 lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures 1471:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 8 76 theme healthy 1691:1697 arg1 cells 1699:1703 healthy cells 1691:1703 healthy cells 1691:1703 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 13 77 theme branched 2383:2390 arg1 structures 2399:2408 branched core 2 structures 2383:2408 branched core 2 structures 2383:2408 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 4 78 theme current 638:644 arg1 work 646:649 the current work 634:649 the current work 634:649 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 3 79 theme healthy 585:591 arg1 cells 593:597 healthy cells 585:597 healthy cells 585:597 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 7 80 theme branched 1492:1499 arg1 structures 1508:1517 the branched core 2 structures 1488:1517 the branched core 2 structures 1488:1517 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 8 81 located found 1682:1686 arg2 structures 1658:1667 The core 2 structures 1647:1667 The core 2 structures 1647:1667 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 8 81 located found 1682:1686 arg1 cells 1699:1703 healthy cells 1691:1703 healthy cells 1691:1703 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 5 82 gly glycosylated 1178:1189 arg2 sites 1233:1237 the five different sites 1214:1237 the five different sites of the MUC1 tandem repeat 1214:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 82 gly glycosylated 1178:1189 arg1 type-1 1106:1111 type-1 1106:1111 type-1 1106:1111 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 82 gly glycosylated 1178:1189 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 82 gly glycosylated 1178:1189 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 82 gly glycosylated 1178:1189 arg2 repeat 1258:1263 the MUC1 tandem repeat 1242:1263 the MUC1 tandem repeat 1242:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 4 83 theme different 708:716 arg1 stimulants 725:734 different immune stimulants 708:734 different immune stimulants 708:734 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 2 84 gly glycopeptide 305:316 arg2 glycopeptide 305:316 tumor-associated glycopeptide structures 288:327 tumor-associated glycopeptide structures 288:327 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. 27957769 10 85 theme induced 2000:2006 arg1 antibodies 2008:2017 the induced antibodies 1996:2017 the induced antibodies 1996:2017 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 11 86 theme antibody 2108:2115 arg1 response 2117:2124 the antibody response 2104:2124 the antibody response 2104:2124 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 13 87 theme core 2 2392:2397 arg1 structures 2399:2408 branched core 2 structures 2383:2408 branched core 2 structures 2383:2408 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 4 88 theme vaccine 673:679 arg1 candidates 681:690 three MUC1 antitumor vaccine candidates 652:690 three MUC1 antitumor vaccine candidates conjugated with different immune stimulants 652:734 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 13 89 located found 2421:2425 arg1 cells 2438:2442 healthy cells 2430:2442 healthy cells 2430:2442 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 13 89 located found 2421:2425 arg2 structures 2399:2408 branched core 2 structures 2383:2408 branched core 2 structures 2383:2408 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 5 90 theme elongated 1124:1132 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 90 theme elongated 1124:1132 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 9 91 gly glycopeptide 1941:1952 arg2 glycopeptide 1941:1952 the branched core 2 glycopeptide epitopes 1921:1961 the branched core 2 glycopeptide epitopes 1921:1961 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 8 92 theme synthetic 1819:1827 arg1 vaccines 1829:1836 synthetic vaccines 1819:1836 synthetic vaccines 1819:1836 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27957769 10 93 theme glycosylation 2058:2070 arg1 specificity 2077:2087 very high glycosylation site specificity 2048:2087 very high glycosylation site specificity 2048:2087 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 11 94 theme glycosylation 2185:2197 arg1 sites 2199:2203 glycosylation sites 2185:2203 glycosylation sites in either the immune-dominant PDTR or the GSTA domain 2185:2257 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 2 95 theme tumor-associated 288:303 arg1 structures 318:327 tumor-associated glycopeptide structures 288:327 tumor-associated glycopeptide structures 288:327 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. 27957769 5 96 theme antibody 943:950 arg1 profiling 963:971 antibody microarray profiling 943:971 antibody microarray profiling 943:971 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 7 97 theme antigen 1627:1633 arg1 structures 1635:1644 truncated tumor-associated antigen structures 1600:1644 truncated tumor-associated antigen structures 1600:1644 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 1 98 theme vaccine 165:171 arg1 development 173:183 vaccine development 165:183 vaccine development 165:183 Glycoprotein research is pivotal for vaccine development and biomarker discovery. 27957769 7 99 theme truncated 1600:1608 arg1 structures 1635:1644 truncated tumor-associated antigen structures 1600:1644 truncated tumor-associated antigen structures 1600:1644 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 5 100 dep glycans 1014:1020 arg1 STN 1028:1030 TN , STN , and T-antigen structures 1023:1057 STN 1028:1030 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 100 dep glycans 1014:1020 arg1 structures 1048:1057 TN , STN , and T-antigen structures 1023:1057 structures 1048:1057 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 0 101 theme Mucin 105:109 arg1 Structures 116:125 Diverse Mucin Core Structures 97:125 Diverse Mucin Core Structures 97:125 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 13 102 theme antisera 2483:2490 arg1 recognition 2464:2474 antibody recognition 2455:2474 antibody recognition of the antisera 2455:2490 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 13 103 with glycosylation 2364:2376 arg1 structures 2399:2408 branched core 2 structures 2383:2408 branched core 2 structures 2383:2408 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 13 104 theme humoral 2572:2578 arg1 response 2587:2594 a tumor-specific humoral immune response 2555:2594 a tumor-specific humoral immune response 2555:2594 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 11 105 theme response 2117:2124 arg1 different 2148:2156 different 2148:2156 different 2148:2156 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 11 105 theme response 2117:2124 arg1 nature 2094:2099 The nature 2090:2099 The nature of the antibody response 2090:2124 The nature of the antibody response is characteristically different for antibodies directed to glycosylation sites in either the immune-dominant PDTR or the GSTA domain. 27957769 9 106 theme extended 1848:1855 arg1 structures 1868:1877 the extended mucin core structures 1844:1877 the extended mucin core structures in hand 1844:1885 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 5 107 theme immune 806:811 arg1 stimulant 813:821 the immune stimulant 802:821 the immune stimulant 802:821 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 10 108 gly glycosylation 2058:2070 arg2 site 2072:2075 very high glycosylation site specificity 2048:2087 very high glycosylation site specificity 2048:2087 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 10 108 gly glycosylation 2058:2070 arg2 specificity 2077:2087 very high glycosylation site specificity 2048:2087 very high glycosylation site specificity 2048:2087 It is observed that the induced antibodies recognize MUC1 peptides with very high glycosylation site specificity. 27957769 7 109 theme tumor 1361:1365 arg1 cells 1367:1371 tumor cells 1361:1371 tumor cells 1361:1371 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 3 110 theme raised 405:410 arg1 reactions 432:440 the raised polyclonal, humoral reactions 401:440 the raised polyclonal, humoral reactions 401:440 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 5 111 theme mucin 875:879 arg1 glycopeptides 883:895 mucin 1 glycopeptides 875:895 mucin 1 glycopeptides (>100 entries) 875:910 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 111 theme mucin 875:879 arg1 entries 903:909 >100 entries 898:909 >100 entries 898:909 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 112 theme tumor-associated 997:1012 arg1 glycans 1014:1020 small tumor-associated glycans 991:1020 small tumor-associated glycans (TN , STN , and T-antigen structures) 991:1058 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 0 113 theme Antibodies 23:32 arg1 Analysis 11:18 Microarray Analysis 0:18 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 0:126 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 3 114 theme reactions 432:440 arg1 specificity 386:396 specificity 386:396 specificity 386:396 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 3 114 theme reactions 432:440 arg1 quality 374:380 quality 374:380 quality 374:380 Deeper insights into the quality and specificity of the raised polyclonal, humoral reactions are often not addressed, despite the fact that an immunological memory, which produces antibodies with cross-reactivity to epitopes exposed on healthy cells, may cause autoimmune diseases. 27957769 12 115 theme saccharide 2323:2332 arg1 structures 2334:2343 the tumor-associated saccharide structures 2302:2343 the tumor-associated saccharide structures on MUC1 2302:2351 All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. 27957769 13 116 theme antibody 2455:2462 arg1 recognition 2464:2474 antibody recognition 2455:2474 antibody recognition of the antisera 2455:2490 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 13 117 theme Extensive 2354:2362 arg1 glycosylation 2364:2376 Extensive glycosylation 2354:2376 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, 2354:2443 Extensive glycosylation with branched core 2 structures, typically found on healthy cells, abolishes antibody recognition of the antisera and suggests that all vaccine conjugates preferentially induce a tumor-specific humoral immune response. 27957769 0 118 theme Core 111:114 arg1 Structures 116:125 Diverse Mucin Core Structures 97:125 Diverse Mucin Core Structures 97:125 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 0 119 theme Antitumor 57:65 arg1 Vaccines 67:74 Synthetic Antitumor Vaccines 47:74 Synthetic Antitumor Vaccines 47:74 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 5 120 theme variable 1194:1201 arg1 density 1203:1209 variable density 1194:1209 variable density 1194:1209 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 121 theme O-glycan 1080:1087 arg1 type-1 1106:1111 type-1 1106:1111 type-1 1106:1111 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 121 theme O-glycan 1080:1087 arg1 structures 1094:1103 heavily extended O-glycan core structures 1063:1103 heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat 1063:1263 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 121 theme O-glycan 1080:1087 arg1 tri- 1143:1146 type-2 elongated core 1-3 tri- 1117:1146 type-2 elongated core 1-3 tri- 1117:1146 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 122 gly glycopeptides 883:895 arg2 glycopeptides 883:895 mucin 1 glycopeptides 875:895 mucin 1 glycopeptides (>100 entries) 875:910 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 5 122 gly glycopeptides 883:895 arg2 entries 903:909 >100 entries 898:909 >100 entries 898:909 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 7 123 theme core 3 1562:1567 arg1 structures 1569:1578 core 3 structures 1562:1578 core 3 structures 1562:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 5 124 theme T-antigen 1038:1046 arg1 structures 1048:1057 TN , STN , and T-antigen structures 1023:1057 structures 1048:1057 For assessment of the influence of the immune stimulant on antibody recognition, a comprehensive library of mucin 1 glycopeptides (>100 entries) is synthesized and employed in antibody microarray profiling; these range from small tumor-associated glycans (TN , STN , and T-antigen structures) to heavily extended O-glycan core structures (type-1 and type-2 elongated core 1-3 tri-, tetra-, and hexasaccharides) glycosylated in variable density at the five different sites of the MUC1 tandem repeat. 27957769 9 125 theme core 2 1934:1939 arg1 epitopes 1954:1961 the branched core 2 glycopeptide epitopes 1921:1961 the branched core 2 glycopeptide epitopes 1921:1961 With the extended mucin core structures in hand, antibody cross-reactivity toward the branched core 2 glycopeptide epitopes is explored. 27957769 2 126 theme successful 215:224 arg1 methodologies 226:238 Many successful methodologies 210:238 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures 210:327 Many successful methodologies for reliably increasing the antigenicity toward tumor-associated glycopeptide structures have been reported. 27957769 7 127 theme core 1 1552:1557 arg1 formation 1532:1540 formation 1532:1540 formation of linear core 1 or core 3 structures 1532:1578 On tumor cells, the core 2 β-1,6-N-acetylglucosaminyltransferase-1 (C2GlcNAcT-1) is down-regulated, resulting in lower amounts of the branched core 2 structures, which favor formation of linear core 1 or core 3 structures, and in particular, truncated tumor-associated antigen structures. 27957769 0 128 dep Analysis 11:18 arg1 Specificity 77:87 Specificity 77:87 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 0:126 Microarray Analysis of Antibodies Induced with Synthetic Antitumor Vaccines: Specificity against Diverse Mucin Core Structures. 27957769 4 129 theme antitumor 663:671 arg1 candidates 681:690 three MUC1 antitumor vaccine candidates 652:690 three MUC1 antitumor vaccine candidates conjugated with different immune stimulants 652:734 In the current work, three MUC1 antitumor vaccine candidates conjugated with different immune stimulants are evaluated immunologically. 27957769 12 130 theme antibody 2264:2271 arg1 sera 2273:2276 All antibody sera 2260:2276 All antibody sera 2260:2276 All antibody sera show high reactivity to the tumor-associated saccharide structures on MUC1. 27957769 8 131 theme cross-reactivity 1737:1752 arg1 elucidation 1713:1723 the elucidation 1709:1723 the elucidation of antibody cross-reactivity to such epitopes 1709:1769 The core 2 structures are commonly found on healthy cells and the elucidation of antibody cross-reactivity to such epitopes may predict the tumor-selectivity and safety of synthetic vaccines. 27629418 7 0 theme W251N 1187:1191 arg1 mutants 1193:1199 W251N mutants 1187:1199 W251N mutants 1187:1199 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 10 1 theme glycoproteins 1879:1891 arg1 glycoforms 1848:1857 homogeneous glycoforms 1836:1857 homogeneous glycoforms of core-fucosylated glycoproteins 1836:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 5 2 from activity 1019:1026 arg1 form 1048:1051 the afucosylated form 1031:1051 the afucosylated form 1031:1051 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 9 3 theme glycosynthase-like 1529:1546 arg1 activity 1548:1555 a glycosynthase-like activity 1527:1555 a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products 1527:1658 Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. 27629418 10 4 theme substrate 1708:1716 arg1 mechanism 1730:1738 the substrate recognition mechanism 1704:1738 the substrate recognition mechanism of glycoside hydrolase family 85 enzymes 1704:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 10 5 theme glycoprotein-based 1922:1939 arg1 therapeutics 1941:1952 potent glycoprotein-based therapeutics 1915:1952 potent glycoprotein-based therapeutics 1915:1952 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 4 6 theme substrate 869:877 arg1 specificities 879:891 altered substrate specificities 861:891 altered substrate specificities 861:891 Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. 27629418 1 7 contain having 297:302 arg2 glycoform 324:332 a precisely defined glycoform 304:332 a precisely defined glycoform 304:332 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 7 contain having 297:302 arg1 therapeutics 284:295 glycoprotein-based therapeutics 265:295 glycoprotein-based therapeutics having a precisely defined glycoform 265:332 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 5 8 theme mutant 899:904 arg1 enzymes 906:912 Such mutant enzymes 894:912 Such mutant enzymes 894:912 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 10 9 theme glycoside 1743:1751 arg1 family 1763:1768 glycoside hydrolase family 85 1743:1771 glycoside hydrolase family 85 enzymes 1743:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 5 10 gly α1,6-fucosylated 960:975 arg1 structure 994:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 1 11 theme useful 194:199 arg1 M 126:126 Endo-β-N-acetylglucosaminidase M 95:126 Endo-β-N-acetylglucosaminidase M (Endo-M) 95:135 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 11 theme useful 194:199 arg1 tool 201:204 a useful tool 192:204 a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity 192:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 10 12 theme enzymes 1773:1779 arg1 mechanism 1730:1738 the substrate recognition mechanism 1704:1738 the substrate recognition mechanism of glycoside hydrolase family 85 enzymes 1704:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 4 13 theme mutant 739:744 arg1 enzymes 746:752 the Endo-M mutant enzymes 728:752 the Endo-M mutant enzymes generated 728:762 Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. 27629418 10 14 theme family 1763:1768 arg1 enzymes 1773:1779 glycoside hydrolase family 85 enzymes 1743:1779 glycoside hydrolase family 85 enzymes 1743:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 5 15 theme α1,6-fucosylated 960:975 arg1 structure 994:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 8 16 theme substrate 1392:1400 arg1 intermediate 1402:1413 an activated substrate intermediate 1379:1413 an activated substrate intermediate (sialyl glyco-oxazoline) 1379:1438 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 8 16 theme substrate 1392:1400 arg1 glyco-oxazoline 1423:1437 sialyl glyco-oxazoline 1416:1437 sialyl glyco-oxazoline 1416:1437 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 9 17 theme transglycosylation 1632:1649 arg1 products 1651:1658 the transglycosylation products 1628:1658 the transglycosylation products 1628:1658 Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. 27629418 5 18 gly afucosylated 1035:1046 arg1 form 1048:1051 the afucosylated form 1031:1051 the afucosylated form 1031:1051 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 0 19 theme Core-fucosylated 67:82 arg1 N-Glycans 84:92 Core-fucosylated N-Glycans 67:92 Core-fucosylated N-Glycans 67:92 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 1 20 theme transglycosylation 352:369 arg1 activity 371:378 its transglycosylation activity 348:378 its transglycosylation activity 348:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 4 21 from position 798:805 arg1 tryptophan 784:793 the tryptophan 780:793 the tryptophan at position 251 780:809 Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. 27629418 1 22 theme fungus 166:171 arg1 hiemalis 179:186 the fungus Mucor hiemalis 162:186 the fungus Mucor hiemalis 162:186 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 7 23 theme lactoferrin 1284:1294 arg1 glycoproteins 1296:1308 human lactoferrin glycoproteins 1278:1308 human lactoferrin glycoproteins 1278:1308 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 5 24 theme core 989:992 arg1 structure 994:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 8 25 theme α1,6-fucosyl-N-acetylglucosaminyl 1448:1480 arg1 biotin 1482:1487 an α1,6-fucosyl-N-acetylglucosaminyl biotin 1445:1487 an α1,6-fucosyl-N-acetylglucosaminyl biotin 1445:1487 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 5 26 theme afucosylated 1035:1046 arg1 form 1048:1051 the afucosylated form 1031:1051 the afucosylated form 1031:1051 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 6 27 gly core-fucosylated 1159:1174 arg1 substrate 1176:1184 the core-fucosylated substrate 1155:1184 the core-fucosylated substrate 1155:1184 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 7 28 theme core-fucosylated 1243:1258 arg1 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 1 29 theme Endo-β-N-acetylglucosaminidase 95:124 arg1 tool 201:204 a useful tool 192:204 a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity 192:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 29 theme Endo-β-N-acetylglucosaminidase 95:124 arg1 Endo-M 129:134 Endo-M 129:134 Endo-M 129:134 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 29 theme Endo-β-N-acetylglucosaminidase 95:124 arg1 M 126:126 Endo-β-N-acetylglucosaminidase M 95:126 Endo-β-N-acetylglucosaminidase M (Endo-M) 95:135 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 29 theme Endo-β-N-acetylglucosaminidase 95:124 arg1 endoglycosidase 141:155 an endoglycosidase 138:155 an endoglycosidase from the fungus Mucor hiemalis 138:186 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 6 30 theme W251N 1108:1112 arg1 mutant 1114:1119 the W251N mutant 1104:1119 the W251N mutant 1104:1119 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 6 30 theme W251N 1108:1112 arg1 efficient 1130:1138 efficient 1130:1138 efficient 1130:1138 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 7 31 theme immunoglobulin 1218:1231 arg1 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 5 32 theme increased 924:932 arg1 hydrolysis 934:943 increased hydrolysis 924:943 increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure 924:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 2 33 theme various 422:428 arg1 N-glycans 430:438 various N-glycans 422:438 various N-glycans 422:438 Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. 27629418 10 34 theme application 1811:1821 arg1 scope 1802:1806 their scope 1796:1806 their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics 1796:1952 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 1 35 theme Mucor 173:177 arg1 hiemalis 179:186 the fungus Mucor hiemalis 162:186 the fungus Mucor hiemalis 162:186 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 0 36 theme Mucor 23:27 arg1 Endoglycosidase 38:52 a Mutant Mucor hiemalis Endoglycosidase 14:52 a Mutant Mucor hiemalis Endoglycosidase That 14:57 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 0 37 dep Endoglycosidase 38:52 arg1 That 54:57 That 54:57 That 54:57 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 2 38 theme core-fucosylated 460:475 arg1 N-glycans 477:485 core-fucosylated N-glycans 460:485 core-fucosylated N-glycans 460:485 Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. 27629418 10 39 theme core-fucosylated 1862:1877 arg1 glycoproteins 1879:1891 core-fucosylated glycoproteins 1862:1891 core-fucosylated glycoproteins 1862:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 0 40 theme Endoglycosidase 38:52 arg1 Generation 0:9 Generation 0:9 Generation of a Mutant Mucor hiemalis Endoglycosidase That 0:57 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 2 41 gly core-fucosylated 460:475 arg1 N-glycans 477:485 core-fucosylated N-glycans 460:485 core-fucosylated N-glycans 460:485 Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. 27629418 3 42 theme core-α1,6-fucosylated 691:711 arg1 glycans 713:719 mammalian-type core-α1,6-fucosylated glycans 676:719 mammalian-type core-α1,6-fucosylated glycans 676:719 Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. 27629418 7 43 gly core-fucosylated 1243:1258 arg1 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 1 44 theme chemoenzymatic 210:223 arg1 synthesis 225:233 chemoenzymatic synthesis 210:233 chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity 210:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 6 45 theme core-fucosylated 1159:1174 arg1 substrate 1176:1184 the core-fucosylated substrate 1155:1184 the core-fucosylated substrate 1155:1184 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 6 46 from efficient 1130:1138 arg1 addition 1067:1074 addition 1067:1074 addition 1067:1074 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 9 47 theme N175Q 1564:1568 arg1 substitution 1570:1581 a N175Q substitution 1562:1581 a N175Q substitution 1562:1581 Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. 27629418 10 48 theme recognition 1718:1728 arg1 mechanism 1730:1738 the substrate recognition mechanism 1704:1738 the substrate recognition mechanism of glycoside hydrolase family 85 enzymes 1704:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 10 49 theme therapeutics 1941:1952 arg1 production 1901:1910 the production 1897:1910 the production of potent glycoprotein-based therapeutics 1897:1952 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 1 50 theme glycoprotein-based 265:282 arg1 therapeutics 284:295 glycoprotein-based therapeutics 265:295 glycoprotein-based therapeutics having a precisely defined glycoform 265:332 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 10 51 theme potent 1915:1920 arg1 therapeutics 1941:1952 potent glycoprotein-based therapeutics 1915:1952 potent glycoprotein-based therapeutics 1915:1952 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 5 52 theme Such 894:897 arg1 enzymes 906:912 Such mutant enzymes 894:912 Such mutant enzymes 894:912 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 4 53 theme altered 861:867 arg1 specificities 879:891 altered substrate specificities 861:891 altered substrate specificities 861:891 Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. 27629418 10 54 theme hydrolase 1753:1761 arg1 family 1763:1768 glycoside hydrolase family 85 1743:1771 glycoside hydrolase family 85 enzymes 1743:1779 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 10 55 gly glycoproteins 1879:1891 arg1 glycoproteins 1879:1891 core-fucosylated glycoproteins 1862:1891 core-fucosylated glycoproteins 1862:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 8 56 theme sialyl 1360:1365 arg1 glycan 1367:1372 the sialyl glycan 1356:1372 the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) 1356:1438 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 3 57 theme site-directed 590:602 arg1 mutagenesis 604:614 site-directed mutagenesis 590:614 site-directed mutagenesis 590:614 Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. 27629418 5 58 theme synthetic 950:958 arg1 structure 994:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 4 59 theme Endo-M 732:737 arg1 enzymes 746:752 the Endo-M mutant enzymes 728:752 the Endo-M mutant enzymes generated 728:762 Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. 27629418 3 60 theme mutant 637:642 arg1 enzymes 644:650 mutant enzymes 637:650 mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans 637:719 Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. 27629418 5 61 theme trimannosyl 977:987 arg1 structure 994:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 a synthetic α1,6-fucosylated trimannosyl core structure 948:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 8 62 theme activated 1382:1390 arg1 intermediate 1402:1413 an activated substrate intermediate 1379:1413 an activated substrate intermediate (sialyl glyco-oxazoline) 1379:1438 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 8 62 theme activated 1382:1390 arg1 glyco-oxazoline 1423:1437 sialyl glyco-oxazoline 1416:1437 sialyl glyco-oxazoline 1416:1437 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 7 63 link G-derived 1233:1241 arg1 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 9 64 theme products 1651:1658 arg1 accumulation 1612:1623 accumulation 1612:1623 accumulation of the transglycosylation products 1612:1658 Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. 27629418 1 65 from hiemalis 179:186 arg1 M 126:126 Endo-β-N-acetylglucosaminidase M 95:126 Endo-β-N-acetylglucosaminidase M (Endo-M) 95:135 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 1 65 from hiemalis 179:186 arg1 endoglycosidase 141:155 an endoglycosidase 138:155 an endoglycosidase from the fungus Mucor hiemalis 138:186 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 6 66 from addition 1067:1074 arg1 mutant 1114:1119 the W251N mutant 1104:1119 the W251N mutant 1104:1119 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 6 66 from addition 1067:1074 arg1 efficient 1130:1138 efficient 1130:1138 efficient 1130:1138 In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. 27629418 8 67 theme sialyl 1416:1421 arg1 intermediate 1402:1413 an activated substrate intermediate 1379:1413 an activated substrate intermediate (sialyl glyco-oxazoline) 1379:1438 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 8 67 theme sialyl 1416:1421 arg1 glyco-oxazoline 1423:1437 sialyl glyco-oxazoline 1416:1437 sialyl glyco-oxazoline 1416:1437 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 7 68 theme human 1278:1282 arg1 glycoproteins 1296:1308 human lactoferrin glycoproteins 1278:1308 human lactoferrin glycoproteins 1278:1308 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 1 69 theme activity 371:378 arg1 virtue 338:343 virtue 338:343 virtue of its transglycosylation activity 338:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 10 70 gly core-fucosylated 1862:1877 arg1 glycoproteins 1879:1891 core-fucosylated glycoproteins 1862:1891 core-fucosylated glycoproteins 1862:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 0 71 gly Core-fucosylated 67:82 arg1 N-Glycans 84:92 Core-fucosylated N-Glycans 67:92 Core-fucosylated N-Glycans 67:92 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 2 72 theme mammalian 510:518 arg1 glycoproteins 520:532 mammalian glycoproteins 510:532 mammalian glycoproteins 510:532 Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. 27629418 7 73 gly glycopeptides 1260:1272 arg2 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 7 74 theme G-derived 1233:1241 arg1 glycopeptides 1260:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 the immunoglobulin G-derived core-fucosylated glycopeptides 1214:1272 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 10 75 gly glycoforms 1848:1857 arg1 glycoproteins 1879:1891 core-fucosylated glycoproteins 1862:1891 core-fucosylated glycoproteins 1862:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 5 76 theme structure 994:1002 arg1 hydrolysis 934:943 increased hydrolysis 924:943 increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure 924:1002 Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. 27629418 0 77 theme Mutant 16:21 arg1 Endoglycosidase 38:52 a Mutant Mucor hiemalis Endoglycosidase 14:52 a Mutant Mucor hiemalis Endoglycosidase That 14:57 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 8 78 from intermediate 1402:1413 arg1 glycan 1367:1372 the sialyl glycan 1356:1372 the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) 1356:1438 This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. 27629418 7 79 gly glycoproteins 1296:1308 arg1 glycoproteins 1296:1308 human lactoferrin glycoproteins 1278:1308 human lactoferrin glycoproteins 1278:1308 W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. 27629418 0 80 theme hiemalis 29:36 arg1 Endoglycosidase 38:52 a Mutant Mucor hiemalis Endoglycosidase 14:52 a Mutant Mucor hiemalis Endoglycosidase That 14:57 Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans. 27629418 1 81 theme defined 316:322 arg1 glycoform 324:332 a precisely defined glycoform 304:332 a precisely defined glycoform 304:332 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 27629418 3 82 theme mammalian-type 676:689 arg1 glycans 713:719 mammalian-type core-α1,6-fucosylated glycans 676:719 mammalian-type core-α1,6-fucosylated glycans 676:719 Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. 27629418 3 83 gly core-α1,6-fucosylated 691:711 arg1 glycans 713:719 mammalian-type core-α1,6-fucosylated glycans 676:719 mammalian-type core-α1,6-fucosylated glycans 676:719 Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. 27629418 2 84 gly glycoproteins 520:532 arg1 glycoproteins 520:532 mammalian glycoproteins 510:532 mammalian glycoproteins 510:532 Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. 27629418 10 85 theme homogeneous 1836:1846 arg1 glycoforms 1848:1857 homogeneous glycoforms 1836:1857 homogeneous glycoforms of core-fucosylated glycoproteins 1836:1891 These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. 27629418 9 86 theme W251N 1507:1511 arg1 mutant 1513:1518 the W251N mutant 1503:1518 the W251N mutant 1503:1518 Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. 27629418 1 87 theme glycoconjugates 238:252 arg1 synthesis 225:233 chemoenzymatic synthesis 210:233 chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity 210:378 Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. 28509333 6 0 theme respective 1031:1040 arg1 lectin 1042:1047 the respective lectin 1027:1047 the respective lectin 1027:1047 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 4 1 with Patients 681:688 arg1 RA 708:709 recent-onset RA 695:709 recent-onset RA at baseline and after 3-year follow-up 695:748 Patients with recent-onset RA at baseline and after 3-year follow-up were investigated. 28509333 3 2 theme fucosyl 488:494 arg1 AAL 506:508 AAL 506:508 AAL 506:508 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 2 theme fucosyl 488:494 arg1 residues 496:503 fucosyl residues 488:503 fucosyl residues (AAL) 488:509 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 10 3 theme RA 1569:1570 arg1 treatment 1556:1564 successful treatment 1545:1564 successful treatment of RA 1545:1570 Hence, successful treatment of RA was associated with an increase in the SNA/GalNAc-L ratio. 28509333 3 4 theme fucosylated 512:522 arg1 LCA 557:559 LCA 557:559 LCA 557:559 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 4 theme fucosylated 512:522 arg1 sites 550:554 fucosylated tri-mannose N-glycan core sites 512:554 fucosylated tri-mannose N-glycan core sites (LCA) 512:560 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 1 5 theme immune 244:249 arg1 complexes 251:259 serum immune complexes 238:259 serum immune complexes in patients with rheumatoid arthritis (RA) 238:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 12 6 theme glycans 1766:1772 arg1 exposure 1754:1761 the exposure 1750:1761 the exposure of glycans in native complexed IgG of patients with early RA 1750:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 11 7 theme SNA/GalNAc-L 1650:1661 arg1 ratio 1663:1667 the SNA/GalNAc-L ratio 1646:1667 the SNA/GalNAc-L ratio in particular 1646:1681 Interestingly, the SNA/GalNAc-L ratio in particular rises after glucocorticoid treatment. 28509333 5 8 theme native 783:788 arg1 IgG 790:792 native IgG 783:792 native IgG 783:792 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 10 9 from increase 1595:1602 arg1 ratio 1624:1628 the SNA/GalNAc-L ratio 1607:1628 the SNA/GalNAc-L ratio 1607:1628 Hence, successful treatment of RA was associated with an increase in the SNA/GalNAc-L ratio. 28509333 1 10 theme glycosylation 174:186 arg1 profile 188:194 the glycosylation profile 170:194 the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 170:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 3 11 theme N-glycan 536:543 arg1 LCA 557:559 LCA 557:559 LCA 557:559 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 11 theme N-glycan 536:543 arg1 sites 550:554 fucosylated tri-mannose N-glycan core sites 512:554 fucosylated tri-mannose N-glycan core sites (LCA) 512:560 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 9 12 theme acid 1442:1445 arg1 accessibility 1447:1459 sialic acid accessibility 1435:1459 sialic acid accessibility 1435:1459 While sialic acid accessibility increased during treatment, the accessibility of galactose/GalNAc decreased. 28509333 12 13 from exposure 1754:1761 arg1 IgG 1794:1796 native complexed IgG 1777:1796 native complexed IgG of patients with early RA 1777:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 10 14 theme SNA/GalNAc-L 1611:1622 arg1 ratio 1624:1628 the SNA/GalNAc-L ratio 1607:1628 the SNA/GalNAc-L ratio 1607:1628 Hence, successful treatment of RA was associated with an increase in the SNA/GalNAc-L ratio. 28509333 1 15 theme immunoglobulin 206:219 arg1 G 225:225 native immunoglobulin (Ig)G 199:225 native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 199:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 8 16 with patients 1399:1406 arg1 RA 1413:1414 RA 1413:1414 RA 1413:1414 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 4 17 theme recent-onset 695:706 arg1 RA 708:709 recent-onset RA 695:709 recent-onset RA at baseline and after 3-year follow-up 695:748 Patients with recent-onset RA at baseline and after 3-year follow-up were investigated. 28509333 1 18 attach present 227:233 arg1 complexes 251:259 serum immune complexes 238:259 serum immune complexes in patients with rheumatoid arthritis (RA) 238:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 1 18 attach present 227:233 arg2 G 225:225 native immunoglobulin (Ig)G 199:225 native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 199:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 6 19 theme total 956:960 arg1 higher 1067:1072 higher 1067:1072 higher 1067:1072 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 6 19 theme total 956:960 arg1 accessibility 962:974 The total accessibility 952:974 The total accessibility of fucose residues on captured immune complexes to the respective lectin 952:1047 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 3 20 gly fucosylated 512:522 arg1 LCA 557:559 LCA 557:559 LCA 557:559 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 20 gly fucosylated 512:522 arg1 sites 550:554 fucosylated tri-mannose N-glycan core sites 512:554 fucosylated tri-mannose N-glycan core sites (LCA) 512:560 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 21 theme sialic 572:577 arg1 SNA 594:596 SNA 594:596 SNA 594:596 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 21 theme sialic 572:577 arg1 residues 584:591 terminal sialic acid residues 563:591 terminal sialic acid residues (SNA) 563:597 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 0 22 from accessibility 15:27 arg1 complexes 56:64 native immunoglobulin G complexes 32:64 native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy 32:125 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 8 23 theme higher 1291:1296 arg1 accessibility 1298:1310 a significantly higher accessibility 1275:1310 a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA 1275:1414 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 0 24 theme rheumatoid 75:84 arg1 arthritis 86:94 early rheumatoid arthritis 69:94 early rheumatoid arthritis 69:94 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 5 25 theme C-reactive 856:865 arg1 CRP 876:878 CRP 876:878 CRP 876:878 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 5 25 theme C-reactive 856:865 arg1 protein 867:873 C-reactive protein 856:873 C-reactive protein (CRP) 856:879 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 6 26 from accessibility 962:974 arg1 complexes 1014:1022 captured immune complexes 998:1022 captured immune complexes 998:1022 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 4 27 theme 3-year 733:738 arg1 follow-up 740:748 3-year follow-up 733:748 3-year follow-up 733:748 Patients with recent-onset RA at baseline and after 3-year follow-up were investigated. 28509333 8 28 theme acid 1322:1325 arg1 residues 1327:1334 sialic acid residues 1315:1334 sialic acid residues 1315:1334 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 7 29 theme immune 1144:1149 arg1 complexes 1151:1159 IgG-containing immune complexes 1129:1159 IgG-containing immune complexes 1129:1159 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 9 30 theme galactose/GalNAc 1510:1525 arg1 accessibility 1493:1505 the accessibility 1489:1505 the accessibility of galactose/GalNAc 1489:1525 While sialic acid accessibility increased during treatment, the accessibility of galactose/GalNAc decreased. 28509333 0 31 theme Altered 0:6 arg1 accessibility 15:27 Altered glycan accessibility 0:27 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 0:126 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 5 32 from protein 867:873 arg1 patients 887:894 RA patients 884:894 RA patients 884:894 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 5 33 theme RA 884:885 arg1 patients 887:894 RA patients 884:894 RA patients 884:894 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 6 34 theme captured 998:1005 arg1 complexes 1014:1022 captured immune complexes 998:1022 captured immune complexes 998:1022 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 2 35 theme IgG-containing 398:411 arg1 complexes 420:428 IgG-containing immune complexes 398:428 IgG-containing immune complexes 398:428 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 2 36 theme present 387:393 arg1 glycans 379:385 glycans 379:385 glycans present on IgG-containing immune complexes 379:428 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 7 37 theme fucose 1105:1110 arg1 accessibility 1112:1124 fucose accessibility 1105:1124 fucose accessibility on IgG-containing immune complexes 1105:1159 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 6 38 theme residues 986:993 arg1 higher 1067:1072 higher 1067:1072 higher 1067:1072 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 6 38 theme residues 986:993 arg1 accessibility 962:974 The total accessibility 952:974 The total accessibility of fucose residues on captured immune complexes to the respective lectin 952:1047 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 8 39 theme native 1375:1380 arg1 IgG 1392:1394 native complexed IgG 1375:1394 native complexed IgG 1375:1394 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 0 40 theme native 32:37 arg1 complexes 56:64 native immunoglobulin G complexes 32:64 native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy 32:125 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 1 41 theme rheumatoid 278:287 arg1 arthritis 289:297 rheumatoid arthritis 278:297 rheumatoid arthritis (RA) 278:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 1 41 theme rheumatoid 278:287 arg1 RA 300:301 RA 300:301 RA 300:301 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 2 42 theme biotinylated 433:444 arg1 lectins 446:452 biotinylated lectins 433:452 biotinylated lectins 433:452 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 0 43 theme G 54:54 arg1 complexes 56:64 native immunoglobulin G complexes 32:64 native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy 32:125 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 8 44 theme patients 1399:1406 arg1 accessibility 1298:1310 a significantly higher accessibility 1275:1310 a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA 1275:1414 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 12 45 theme particular 1835:1844 arg1 patterns 1860:1867 particular glycosylation patterns 1835:1867 particular glycosylation patterns 1835:1867 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 5 46 theme structure 934:942 arg1 alterations 908:918 alterations 908:918 alterations of the native structure of IgG 908:949 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 12 47 theme complexed 1784:1792 arg1 IgG 1794:1796 native complexed IgG 1777:1796 native complexed IgG of patients with early RA 1777:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 10 48 theme successful 1545:1554 arg1 treatment 1556:1564 successful treatment 1545:1564 successful treatment of RA 1545:1570 Hence, successful treatment of RA was associated with an increase in the SNA/GalNAc-L ratio. 28509333 3 49 theme linked 620:625 arg1 GalNac-L 660:667 GalNac-L 660:667 GalNac-L 660:667 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 49 theme linked 620:625 arg1 galactose/N-acetylgalactosamine 627:657 O-glycosidically linked galactose/N-acetylgalactosamine 603:657 O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) 603:668 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 2 50 theme binding 332:338 arg1 assays 340:345 lectin binding assays 325:345 lectin binding assays 325:345 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 5 51 theme IgG 947:949 arg1 structure 934:942 the native structure 923:942 the native structure of IgG 923:949 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 7 52 theme cyclic 1216:1221 arg1 peptides 1237:1244 cyclic citrullinated peptides 1216:1244 cyclic citrullinated peptides (anti-CCP) 1216:1255 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 7 52 theme cyclic 1216:1221 arg1 anti-CCP 1247:1254 anti-CCP 1247:1254 anti-CCP 1247:1254 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 1 53 theme present 227:233 arg1 G 225:225 native immunoglobulin (Ig)G 199:225 native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 199:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 6 54 with patients 1077:1084 arg1 RA 1091:1092 RA 1091:1092 RA 1091:1092 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 7 55 theme antibodies 1202:1211 arg1 levels 1192:1197 the levels 1188:1197 the levels of antibodies to cyclic citrullinated peptides (anti-CCP) 1188:1255 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 1 56 gly glycosylation 174:186 arg1 G 225:225 native immunoglobulin (Ig)G 199:225 native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 199:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 8 57 from residues 1327:1334 arg1 IgG 1392:1394 native complexed IgG 1375:1394 native complexed IgG 1375:1394 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 1 58 theme serum 238:242 arg1 complexes 251:259 serum immune complexes 238:259 serum immune complexes in patients with rheumatoid arthritis (RA) 238:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 8 59 dep native 1375:1380 arg1 complexed 1382:1390 complexed 1382:1390 complexed 1382:1390 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 1 60 with patients 264:271 arg1 arthritis 289:297 rheumatoid arthritis 278:297 rheumatoid arthritis (RA) 278:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 1 60 with patients 264:271 arg1 RA 300:301 RA 300:301 RA 300:301 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 1 61 from complexes 251:259 arg1 patients 264:271 patients 264:271 patients with rheumatoid arthritis (RA) 264:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 1 61 from complexes 251:259 arg1 present 227:233 present 227:233 present 227:233 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 3 62 theme core 545:548 arg1 LCA 557:559 LCA 557:559 LCA 557:559 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 62 theme core 545:548 arg1 sites 550:554 fucosylated tri-mannose N-glycan core sites 512:554 fucosylated tri-mannose N-glycan core sites (LCA) 512:560 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 63 theme tri-mannose 524:534 arg1 LCA 557:559 LCA 557:559 LCA 557:559 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 63 theme tri-mannose 524:534 arg1 sites 550:554 fucosylated tri-mannose N-glycan core sites 512:554 fucosylated tri-mannose N-glycan core sites (LCA) 512:560 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 9 64 theme sialic 1435:1440 arg1 accessibility 1447:1459 sialic acid accessibility 1435:1459 sialic acid accessibility 1435:1459 While sialic acid accessibility increased during treatment, the accessibility of galactose/GalNAc decreased. 28509333 1 65 from present 227:233 arg1 complexes 251:259 serum immune complexes 238:259 serum immune complexes in patients with rheumatoid arthritis (RA) 238:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 12 66 with patients 1801:1808 arg1 RA 1821:1822 early RA 1815:1822 early RA 1815:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 1 67 theme native 199:204 arg1 G 225:225 native immunoglobulin (Ig)G 199:225 native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 199:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 3 68 theme acid 579:582 arg1 SNA 594:596 SNA 594:596 SNA 594:596 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 68 theme acid 579:582 arg1 residues 584:591 terminal sialic acid residues 563:591 terminal sialic acid residues (SNA) 563:597 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 0 69 theme early 69:73 arg1 arthritis 86:94 early rheumatoid arthritis 69:94 early rheumatoid arthritis 69:94 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 3 70 theme terminal 563:570 arg1 SNA 594:596 SNA 594:596 SNA 594:596 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 70 theme terminal 563:570 arg1 residues 584:591 terminal sialic acid residues 563:591 terminal sialic acid residues (SNA) 563:597 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 5 71 from C1q 843:845 arg1 patients 887:894 RA patients 884:894 RA patients 884:894 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 5 72 from IgM 838:840 arg1 patients 887:894 RA patients 884:894 RA patients 884:894 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 1 73 theme study 145:149 arg1 goal 132:135 The goal 128:135 The goal of this study 128:149 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 28509333 0 74 from complexes 56:64 arg1 arthritis 86:94 early rheumatoid arthritis 69:94 early rheumatoid arthritis 69:94 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 0 74 from complexes 56:64 arg1 changes 104:110 its changes 100:110 its changes during therapy 100:125 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 7 75 theme IgG-containing 1129:1142 arg1 complexes 1151:1159 IgG-containing immune complexes 1129:1159 IgG-containing immune complexes 1129:1159 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 12 76 theme pharmaceutical 1895:1908 arg1 treatment 1910:1918 pharmaceutical treatment 1895:1918 pharmaceutical treatment 1895:1918 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 5 77 from C3c 848:850 arg1 patients 887:894 RA patients 884:894 RA patients 884:894 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 7 78 from accessibility 1112:1124 arg1 complexes 1151:1159 IgG-containing immune complexes 1129:1159 IgG-containing immune complexes 1129:1159 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 8 79 theme sialic 1315:1320 arg1 residues 1327:1334 sialic acid residues 1315:1334 sialic acid residues 1315:1334 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 0 80 theme glycan 8:13 arg1 accessibility 15:27 Altered glycan accessibility 0:27 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 0:126 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 4 81 from baseline 714:721 arg1 RA 708:709 recent-onset RA 695:709 recent-onset RA at baseline and after 3-year follow-up 695:748 Patients with recent-onset RA at baseline and after 3-year follow-up were investigated. 28509333 6 82 theme immune 1007:1012 arg1 complexes 1014:1022 captured immune complexes 998:1022 captured immune complexes 998:1022 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 2 83 theme immune 413:418 arg1 complexes 420:428 IgG-containing immune complexes 398:428 IgG-containing immune complexes 398:428 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 11 84 theme glucocorticoid 1695:1708 arg1 treatment 1710:1718 glucocorticoid treatment 1695:1718 glucocorticoid treatment 1695:1718 Interestingly, the SNA/GalNAc-L ratio in particular rises after glucocorticoid treatment. 28509333 8 85 theme galactose/GalNAc 1340:1355 arg1 glyco-epitopes 1357:1370 galactose/GalNAc glyco-epitopes 1340:1370 galactose/GalNAc glyco-epitopes 1340:1370 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 2 86 theme glycans 379:385 arg1 accessibility 362:374 the accessibility 358:374 the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins 358:452 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 0 87 theme immunoglobulin 39:52 arg1 complexes 56:64 native immunoglobulin G complexes 32:64 native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy 32:125 Altered glycan accessibility on native immunoglobulin G complexes in early rheumatoid arthritis and its changes during therapy. 28509333 2 88 attach present 387:393 arg1 complexes 420:428 IgG-containing immune complexes 398:428 IgG-containing immune complexes 398:428 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 2 88 attach present 387:393 arg2 glycans 379:385 glycans 379:385 glycans present on IgG-containing immune complexes 379:428 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 6 89 theme fucose 979:984 arg1 residues 986:993 fucose residues 979:993 fucose residues 979:993 The total accessibility of fucose residues on captured immune complexes to the respective lectin was significantly higher in patients with RA. 28509333 12 90 theme patients 1801:1808 arg1 IgG 1794:1796 native complexed IgG 1777:1796 native complexed IgG of patients with early RA 1777:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 11 91 from ratio 1663:1667 arg1 particular 1672:1681 particular 1672:1681 particular 1672:1681 Interestingly, the SNA/GalNAc-L ratio in particular rises after glucocorticoid treatment. 28509333 12 92 theme glycosylation 1846:1858 arg1 patterns 1860:1867 particular glycosylation patterns 1835:1867 particular glycosylation patterns 1835:1867 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 5 93 theme native 927:932 arg1 structure 934:942 the native structure 923:942 the native structure of IgG 923:949 We found that native IgG was complexed significantly more often with IgM, C1q, C3c and C-reactive protein (CRP) in RA patients, suggesting alterations of the native structure of IgG. 28509333 12 94 theme early 1815:1819 arg1 RA 1821:1822 early RA 1815:1822 early RA 1815:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 7 95 theme citrullinated 1223:1235 arg1 peptides 1237:1244 cyclic citrullinated peptides 1216:1244 cyclic citrullinated peptides (anti-CCP) 1216:1255 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 7 95 theme citrullinated 1223:1235 arg1 anti-CCP 1247:1254 anti-CCP 1247:1254 anti-CCP 1247:1254 Moreover, fucose accessibility on IgG-containing immune complexes correlated positively with the levels of antibodies to cyclic citrullinated peptides (anti-CCP). 28509333 3 96 link linked 620:625 arg1 GalNac-L 660:667 GalNac-L 660:667 GalNac-L 660:667 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 3 96 link linked 620:625 arg1 galactose/N-acetylgalactosamine 627:657 O-glycosidically linked galactose/N-acetylgalactosamine 603:657 O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) 603:668 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 12 97 theme native 1777:1782 arg1 IgG 1794:1796 native complexed IgG 1777:1796 native complexed IgG of patients with early RA 1777:1822 In summary, this study shows the exposure of glycans in native complexed IgG of patients with early RA, revealing particular glycosylation patterns and its changes following pharmaceutical treatment. 28509333 3 98 used used 675:678 arg2 Lectins 470:476 Lectins 470:476 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) 470:668 Lectins capturing fucosyl residues (AAL), fucosylated tri-mannose N-glycan core sites (LCA), terminal sialic acid residues (SNA) and O-glycosidically linked galactose/N-acetylgalactosamine (GalNac-L) were used. 28509333 2 99 theme lectin 325:330 arg1 assays 340:345 lectin binding assays 325:345 lectin binding assays 325:345 To accomplish this, lectin binding assays, detecting the accessibility of glycans present on IgG-containing immune complexes by biotinylated lectins, were employed. 28509333 8 100 from glyco-epitopes 1357:1370 arg1 IgG 1392:1394 native complexed IgG 1375:1394 native complexed IgG 1375:1394 We also observed a significantly higher accessibility to sialic acid residues and galactose/GalNAc glyco-epitopes in native complexed IgG of patients with RA at baseline. 28509333 1 101 theme G 225:225 arg1 profile 188:194 the glycosylation profile 170:194 the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA) 170:302 The goal of this study was to investigate the glycosylation profile of native immunoglobulin (Ig)G present in serum immune complexes in patients with rheumatoid arthritis (RA). 27696564 0 0 theme bacterium 68:76 arg1 proteins 47:54 the flagellar filament proteins 24:54 the flagellar filament proteins of the oral bacterium Treponema denticola 24:96 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 4 1 theme bacterial 970:978 arg1 flagellins 980:989 bacterial flagellins 970:989 bacterial flagellins 970:989 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 1 2 theme several 148:154 arg1 bacterium 191:199 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 1 2 theme several 148:154 arg1 spirochetes 156:166 several spirochetes 148:166 several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 148:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 0 3 theme oral 63:66 arg1 bacterium 68:76 the oral bacterium Treponema denticola 59:96 the oral bacterium Treponema denticola 59:96 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 5 4 theme flagellar 1131:1139 arg1 assembly 1141:1148 flagellar assembly 1131:1148 flagellar assembly 1131:1148 Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. 27696564 5 5 theme mutagenesis 1045:1055 arg1 studies 1057:1063 mutagenesis studies 1045:1063 mutagenesis studies 1045:1063 Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. 27696564 3 6 contain has 710:712 arg2 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 6 contain has 710:712 arg2 configuration 733:745 a pseudaminic acid configuration 714:745 a pseudaminic acid configuration (L-glycero-L-manno) 714:765 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 6 contain has 710:712 arg1 Non 706:708 Non 706:708 Non 706:708 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 6 contain has 710:712 arg1 β-linked 774:781 β-linked 774:781 β-linked 774:781 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 7 theme N7 625:626 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 7 theme N7 625:626 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 8 theme denticola 880:888 arg1 FlaBs 867:871 FlaBs 867:871 FlaBs 867:871 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 4 8 theme denticola 880:888 arg1 proteins 857:864 the flagellin proteins 843:864 the flagellin proteins (FlaBs) of T. denticola 843:888 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 1 9 theme pertinent 272:280 arg1 structures 289:298 the pertinent glycan structures 268:298 the pertinent glycan structures 268:298 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 3 10 theme nonulosonic 580:590 arg1 acid 592:595 a monoacetylated nonulosonic acid 563:595 a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 563:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 11 theme flagellins 980:989 arg1 region 960:965 a highly conserved region 941:965 a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5 941:1029 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 4 11 theme flagellins 980:989 arg1 domain 933:938 the D1 domain 926:938 the D1 domain 926:938 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 1 12 theme glycan 282:287 arg1 structures 289:298 the pertinent glycan structures 268:298 the pertinent glycan structures 268:298 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 3 13 theme Da 545:546 arg1 glycan 524:529 The identified glycan 509:529 The identified glycan of mass 450.2 Da 509:546 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 14 theme novel 821:825 arg1 glycan 827:832 This novel glycan 816:832 This novel glycan 816:832 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 3 15 theme pseudaminic 716:726 arg1 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 15 theme pseudaminic 716:726 arg1 configuration 733:745 a pseudaminic acid configuration 714:745 a pseudaminic acid configuration (L-glycero-L-manno) 714:765 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 16 theme flagellin 847:855 arg1 FlaBs 867:871 FlaBs 867:871 FlaBs 867:871 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 4 16 theme flagellin 847:855 arg1 proteins 857:864 the flagellin proteins 843:864 the flagellin proteins (FlaBs) of T. denticola 843:888 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 0 17 theme novel 2:6 arg1 glycan 8:13 A novel glycan 0:13 A novel glycan 0:13 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 3 18 theme acid 728:731 arg1 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 L-glycero-L-manno 748:764 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 18 theme acid 728:731 arg1 configuration 733:745 a pseudaminic acid configuration 714:745 a pseudaminic acid configuration (L-glycero-L-manno) 714:765 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 0 19 dep bacterium 68:76 arg1 denticola 88:96 Treponema denticola 78:96 the oral bacterium Treponema denticola 59:96 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 1 20 theme syphilis 182:189 arg1 bacterium 191:199 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 4 21 theme Toll-like 1010:1018 arg1 receptor 1020:1027 Toll-like receptor 5 1010:1029 Toll-like receptor 5 1010:1029 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 5 22 theme denticola 1169:1177 arg1 motility 1154:1161 motility 1154:1161 motility 1154:1161 Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. 27696564 5 22 theme denticola 1169:1177 arg1 assembly 1141:1148 flagellar assembly 1131:1148 flagellar assembly 1131:1148 Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. 27696564 6 23 theme modification 1231:1242 arg1 sites 1244:1248 its unique modification sites 1220:1248 its unique modification sites 1220:1248 To our knowledge, this novel glycan and its unique modification sites have not been reported previously in any bacteria. 27696564 1 24 gly glycosylation 113:125 arg1 bacterium 191:199 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 178:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 1 24 gly glycosylation 113:125 arg1 spirochetes 156:166 several spirochetes 148:166 several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi 148:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 3 25 dep serine 786:791 arg1 residues 806:813 residues 806:813 residues 806:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 26 with acid 592:595 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 26 with acid 592:595 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 27 theme monoacetylated 565:578 arg1 acid 592:595 a monoacetylated nonulosonic acid 563:595 a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 563:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 28 theme acyl 628:631 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 28 theme acyl 628:631 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 29 theme mass 534:537 arg1 Da 545:546 mass 450.2 Da 534:546 mass 450.2 Da 534:546 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 0 30 theme flagellar 28:36 arg1 proteins 47:54 the flagellar filament proteins 24:54 the flagellar filament proteins of the oral bacterium Treponema denticola 24:96 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 3 31 theme identified 513:522 arg1 glycan 524:529 The identified glycan 509:529 The identified glycan of mass 450.2 Da 509:546 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 32 theme D1 930:931 arg1 region 960:965 a highly conserved region 941:965 a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5 941:1029 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 4 32 theme D1 930:931 arg1 domain 933:938 the D1 domain 926:938 the D1 domain 926:938 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 6 33 theme unique 1224:1229 arg1 sites 1244:1248 its unique modification sites 1220:1248 its unique modification sites 1220:1248 To our knowledge, this novel glycan and its unique modification sites have not been reported previously in any bacteria. 27696564 2 34 theme oral 421:424 arg1 spirochete 426:435 the oral spirochete 417:435 the oral spirochete Treponema denticola 417:455 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 35 theme periodontitis 481:493 arg1 composition 388:398 unusual chemical composition 371:398 unusual chemical composition 371:398 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 35 theme periodontitis 481:493 arg1 pathogen 469:476 a keystone pathogen 458:476 a keystone pathogen of periodontitis 458:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 3 36 attach β-linked 774:781 arg1 threonine 796:804 threonine 796:804 threonine 796:804 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 36 attach β-linked 774:781 arg1 serine 786:791 serine 786:791 serine 786:791 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 36 attach β-linked 774:781 arg2 β-linked 774:781 β-linked 774:781 β-linked 774:781 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 36 attach β-linked 774:781 arg2 Non 706:708 Non 706:708 Non 706:708 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 2 37 with glycan 356:361 arg1 structure 404:412 structure 404:412 structure 404:412 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 37 with glycan 356:361 arg1 composition 388:398 unusual chemical composition 371:398 unusual chemical composition 371:398 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 37 with glycan 356:361 arg1 pathogen 469:476 a keystone pathogen 458:476 a keystone pathogen of periodontitis 458:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 1 38 dep bacterium 191:199 arg1 pallidum 211:218 Treponema pallidum 201:218 Treponema pallidum 201:218 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 1 38 dep bacterium 191:199 arg1 pathogen 237:244 Lyme disease pathogen 224:244 Lyme disease pathogen 224:244 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 1 39 theme Lyme 224:227 arg1 pathogen 237:244 Lyme disease pathogen 224:244 Lyme disease pathogen 224:244 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 2 40 from composition 388:398 arg1 spirochete 426:435 the oral spirochete 417:435 the oral spirochete Treponema denticola 417:455 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 1 41 theme protein 105:111 arg1 glycosylation 113:125 protein glycosylation 105:125 protein glycosylation 105:125 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 0 42 theme filament 38:45 arg1 proteins 47:54 the flagellar filament proteins 24:54 the flagellar filament proteins of the oral bacterium Treponema denticola 24:96 A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola. 27696564 2 43 dep spirochete 426:435 arg1 denticola 447:455 Treponema denticola 437:455 the oral spirochete Treponema denticola 417:455 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 44 theme novel 350:354 arg1 glycan 356:361 a novel glycan 348:361 a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis 348:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 3 45 theme novel 610:614 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 45 theme novel 610:614 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 46 theme multiple 906:913 arg1 sites 915:919 multiple sites 906:919 multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5 906:1029 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 6 47 theme novel 1203:1207 arg1 glycan 1209:1214 this novel glycan 1198:1214 this novel glycan 1198:1214 To our knowledge, this novel glycan and its unique modification sites have not been reported previously in any bacteria. 27696564 3 48 theme extended 616:623 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 48 theme extended 616:623 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 2 49 from structure 404:412 arg1 spirochete 426:435 the oral spirochete 417:435 the oral spirochete Treponema denticola 417:455 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 1 50 theme disease 229:235 arg1 pathogen 237:244 Lyme disease pathogen 224:244 Lyme disease pathogen 224:244 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 3 51 theme 2-methoxy-4,5,6-trihydroxy-hexanoyl 649:683 arg1 residue 685:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue 647:691 a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues 647:813 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 3 51 theme 2-methoxy-4,5,6-trihydroxy-hexanoyl 649:683 arg1 modification 633:644 a novel extended N7 acyl modification 608:644 a novel extended N7 acyl modification 608:644 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 2 52 dep composition 388:398 arg1 an 368:369 an 368:369 an 368:369 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 53 theme unusual 371:377 arg1 composition 388:398 unusual chemical composition 371:398 unusual chemical composition 371:398 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 53 theme unusual 371:377 arg1 pathogen 469:476 a keystone pathogen 458:476 a keystone pathogen of periodontitis 458:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 54 theme chemical 379:386 arg1 composition 388:398 unusual chemical composition 371:398 unusual chemical composition 371:398 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 54 theme chemical 379:386 arg1 pathogen 469:476 a keystone pathogen 458:476 a keystone pathogen of periodontitis 458:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 3 55 dep acid 592:595 arg1 Non 598:600 Non 598:600 Non 598:600 The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. 27696564 4 56 theme conserved 950:958 arg1 region 960:965 a highly conserved region 941:965 a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5 941:1029 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 4 56 theme conserved 950:958 arg1 domain 933:938 the D1 domain 926:938 the D1 domain 926:938 This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. 27696564 1 57 dep pallidum 211:218 arg1 burgdorferi 255:265 Borrelia burgdorferi 246:265 Borrelia burgdorferi 246:265 While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. 27696564 5 58 theme essential 1109:1117 arg1 role 1119:1122 an essential role 1106:1122 an essential role 1106:1122 Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. 27696564 2 59 theme keystone 460:467 arg1 composition 388:398 unusual chemical composition 371:398 unusual chemical composition 371:398 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 27696564 2 59 theme keystone 460:467 arg1 pathogen 469:476 a keystone pathogen 458:476 a keystone pathogen of periodontitis 458:493 Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. 24451126 8 0 theme flexibility 1089:1099 arg1 sites 1130:1134 potential sites 1120:1134 potential sites for engineering antigen binding sites 1120:1172 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 8 0 theme flexibility 1089:1099 arg1 Regions 1067:1073 Regions 1067:1073 Regions of high local flexibility 1067:1099 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 7 1 theme stabilizing 885:895 arg1 effect 897:902 The stabilizing effect 881:902 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 2 theme region 984:989 arg1 impact 964:969 the impact 960:969 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 2 theme region 984:989 arg1 effect 897:902 The stabilizing effect 881:902 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 1 3 theme therapeutic 147:157 arg1 antibodies 170:179 therapeutic monoclonal antibodies 147:179 therapeutic monoclonal antibodies 147:179 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 8 4 theme local 1083:1087 arg1 flexibility 1089:1099 high local flexibility 1078:1099 high local flexibility 1078:1099 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 6 5 theme starting 797:804 arg1 point 806:810 a starting point 795:810 a starting point for simulations of the wild-type protein at two different pH values 795:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 6 5 theme starting 797:804 arg1 structure 721:729 The structure 717:729 The structure of human IgG1-Fc obtained from X-ray crystallography 717:782 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 1 6 theme monoclonal 159:168 arg1 antibodies 170:179 therapeutic monoclonal antibodies 147:179 therapeutic monoclonal antibodies 147:179 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 9 7 from use 1321:1323 arg1 design 1356:1361 the design 1352:1361 the design of antigen binding Fc proteins 1352:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 4 8 theme CH3 571:573 arg1 behavior 542:549 unfolding behavior 532:549 unfolding behavior 532:549 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 4 8 theme CH3 571:573 arg1 stability 518:526 stability 518:526 stability 518:526 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 0 9 theme Fcabs 96:100 arg1 design 86:91 the design 82:91 the design of Fcabs 82:100 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 9 10 theme IgG1-Fc 1252:1258 arg1 approaches 1280:1289 IgG1-Fc, directed evolution approaches 1252:1289 IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1252:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 7 11 theme CH3 938:940 arg1 domain 942:947 the CH3 domain 934:947 the CH3 domain 934:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 12 from effect 897:902 arg1 domain 942:947 the CH3 domain 934:947 the CH3 domain 934:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 9 13 theme antigen 1366:1372 arg1 proteins 1385:1392 antigen binding Fc proteins 1366:1392 antigen binding Fc proteins 1366:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 5 14 theme domains 662:668 arg1 simulations 643:653 molecular dynamics simulations 624:653 molecular dynamics simulations of the domains of the Fc fragment of human IgG1 624:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 7 15 theme mutation 922:929 arg1 impact 964:969 the impact 960:969 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 15 theme mutation 922:929 arg1 effect 897:902 The stabilizing effect 881:902 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 9 16 theme Obtained 1175:1182 arg1 data 1184:1187 Obtained data 1175:1187 Obtained data 1175:1187 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 9 17 theme binding 1374:1380 arg1 proteins 1385:1392 antigen binding Fc proteins 1366:1392 antigen binding Fc proteins 1366:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 6 18 from values 873:878 arg1 point 806:810 a starting point 795:810 a starting point for simulations of the wild-type protein at two different pH values 795:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 6 18 from values 873:878 arg1 structure 721:729 The structure 717:729 The structure of human IgG1-Fc obtained from X-ray crystallography 717:782 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 4 19 dep CH2 563:565 arg1 the 559:561 the 559:561 the 559:561 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 4 19 dep CH2 563:565 arg1 domains 575:581 domains 575:581 domains 575:581 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 7 20 theme CH2 1038:1040 arg1 domains 1042:1048 the CH2 domains 1034:1048 the CH2 domains 1034:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 21 from impact 964:969 arg1 domain 942:947 the CH3 domain 934:947 the CH3 domain 934:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 1 22 theme antibodies 170:179 arg1 development 132:142 the development 128:142 the development of therapeutic monoclonal antibodies 128:179 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 6 23 theme different 860:868 arg1 values 873:878 two different pH values 856:878 two different pH values 856:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 9 24 theme available 1223:1231 arg1 structure 1239:1247 the available X-ray structure 1219:1247 the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1219:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 7 25 theme single 909:914 arg1 mutation 922:929 a single point mutation 907:929 a single point mutation in the CH3 domain 907:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 26 theme hinge 978:982 arg1 region 984:989 the hinge region 974:989 the hinge region 974:989 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 0 27 theme dynamics 10:17 arg1 simulation 19:28 Molecular dynamics simulation 0:28 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 0:101 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 4 28 theme point 485:489 arg1 mutations 491:499 These point mutations 479:499 These point mutations 479:499 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 5 29 theme molecular 624:632 arg1 simulations 643:653 molecular dynamics simulations 624:653 molecular dynamics simulations of the domains of the Fc fragment of human IgG1 624:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 7 30 theme point 916:920 arg1 mutation 922:929 a single point mutation 907:929 a single point mutation in the CH3 domain 907:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 6 31 theme pH 870:871 arg1 values 873:878 two different pH values 856:878 two different pH values 856:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 0 32 theme Molecular 0:8 arg1 simulation 19:28 Molecular dynamics simulation 0:28 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 0:101 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 5 33 theme dynamics 634:641 arg1 simulations 643:653 molecular dynamics simulations 624:653 molecular dynamics simulations of the domains of the Fc fragment of human IgG1 624:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 6 34 theme X-ray 762:766 arg1 crystallography 768:782 X-ray crystallography 762:782 X-ray crystallography 762:782 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 2 35 theme site 320:323 arg1 generation 287:296 generation 287:296 generation of an antigen binding site 287:323 Engineering of its structural loops allows generation of an antigen binding site. 24451126 1 36 theme crystallizable 190:203 arg1 fragment 205:212 the crystallizable fragment 186:212 the crystallizable fragment of IgG1 186:220 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 3 37 theme conformational 368:381 arg1 stability 383:391 the molecule's conformational stability 353:391 the molecule's conformational stability 353:391 However, this might impair the molecule's conformational stability, which can be overcome by introducing stabilizing point mutations in the CH3 domains. 24451126 9 38 dep IgG1-Fc 1252:1258 arg1 directed 1261:1268 directed 1261:1268 directed 1261:1268 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 7 39 theme structure 1011:1019 arg1 impact 964:969 the impact 960:969 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 39 theme structure 1011:1019 arg1 effect 897:902 The stabilizing effect 881:902 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 1 40 from format 118:123 arg1 development 132:142 the development 128:142 the development of therapeutic monoclonal antibodies 128:179 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 2 41 theme binding 312:318 arg1 site 320:323 an antigen binding site 301:323 an antigen binding site 301:323 Engineering of its structural loops allows generation of an antigen binding site. 24451126 3 42 theme CH3 466:468 arg1 domains 470:476 the CH3 domains 462:476 the CH3 domains 462:476 However, this might impair the molecule's conformational stability, which can be overcome by introducing stabilizing point mutations in the CH3 domains. 24451126 7 43 theme glycan 999:1004 arg1 structure 1011:1019 the glycan tree structure 995:1019 the glycan tree structure connected to the CH2 domains 995:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 5 44 theme human 692:696 arg1 IgG1 698:701 human IgG1 692:701 human IgG1 692:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 2 45 theme antigen 304:310 arg1 site 320:323 an antigen binding site 301:323 an antigen binding site 301:323 Engineering of its structural loops allows generation of an antigen binding site. 24451126 9 46 from design 1356:1361 arg1 use 1321:1323 use 1321:1323 use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1321:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 9 46 from design 1356:1361 arg1 stability 1307:1315 stability 1307:1315 stability 1307:1315 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 7 47 from mutation 922:929 arg1 domain 942:947 the CH3 domain 934:947 the CH3 domain 934:947 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 0 48 theme crystallizable 37:50 arg1 fragment 52:59 the crystallizable fragment 33:59 the crystallizable fragment of IgG1-insights 33:76 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 4 49 theme CH2 563:565 arg1 behavior 542:549 unfolding behavior 532:549 unfolding behavior 532:549 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 4 49 theme CH2 563:565 arg1 stability 518:526 stability 518:526 stability 518:526 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 5 50 theme IgG1 698:701 arg1 fragment 680:687 the Fc fragment 673:687 the Fc fragment of human IgG1 673:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 6 51 theme protein 845:851 arg1 simulations 816:826 simulations 816:826 simulations of the wild-type protein 816:851 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 3 52 theme stabilizing 431:441 arg1 mutations 449:457 stabilizing point mutations 431:457 stabilizing point mutations 431:457 However, this might impair the molecule's conformational stability, which can be overcome by introducing stabilizing point mutations in the CH3 domains. 24451126 1 53 theme IgG1 217:220 arg1 fragment 205:212 the crystallizable fragment 186:212 the crystallizable fragment of IgG1 186:220 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 3 54 theme point 443:447 arg1 mutations 449:457 stabilizing point mutations 431:457 stabilizing point mutations 431:457 However, this might impair the molecule's conformational stability, which can be overcome by introducing stabilizing point mutations in the CH3 domains. 24451126 4 55 theme unfolding 532:540 arg1 behavior 542:549 unfolding behavior 532:549 unfolding behavior 532:549 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 7 56 theme tree 1006:1009 arg1 structure 1011:1019 the glycan tree structure 995:1019 the glycan tree structure connected to the CH2 domains 995:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 6 57 theme wild-type 835:843 arg1 protein 845:851 the wild-type protein 831:851 the wild-type protein 831:851 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 8 58 theme antigen 1152:1158 arg1 sites 1168:1172 antigen binding sites 1152:1172 antigen binding sites 1152:1172 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 4 59 dep stability 518:526 arg1 the 514:516 the 514:516 the 514:516 These point mutations often affect the stability and unfolding behavior of both the CH2 and CH3 domains. 24451126 8 60 theme binding 1160:1166 arg1 sites 1168:1172 antigen binding sites 1152:1172 antigen binding sites 1152:1172 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 6 61 theme IgG1-Fc 740:746 arg1 point 806:810 a starting point 795:810 a starting point for simulations of the wild-type protein at two different pH values 795:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 6 61 theme IgG1-Fc 740:746 arg1 structure 721:729 The structure 717:729 The structure of human IgG1-Fc obtained from X-ray crystallography 717:782 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 1 62 theme interesting 106:116 arg1 format 118:123 An interesting format 103:123 An interesting format in the development of therapeutic monoclonal antibodies 103:179 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 0 63 theme fragment 52:59 arg1 simulation 19:28 Molecular dynamics simulation 0:28 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 0:101 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 9 64 theme Fc 1382:1383 arg1 proteins 1385:1392 antigen binding Fc proteins 1366:1392 antigen binding Fc proteins 1366:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 6 65 theme human 734:738 arg1 IgG1-Fc 740:746 human IgG1-Fc 734:746 human IgG1-Fc 734:746 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 9 66 from IgG1-Fc 1341:1347 arg1 design 1356:1361 the design 1352:1361 the design of antigen binding Fc proteins 1352:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 5 67 dep understand 596:605 arg1 to 593:594 to 593:594 to 593:594 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 9 68 theme evolution 1270:1278 arg1 approaches 1280:1289 IgG1-Fc, directed evolution approaches 1252:1289 IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1252:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 5 69 theme Fc 677:678 arg1 fragment 680:687 the Fc fragment 673:687 the Fc fragment of human IgG1 673:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 9 70 from stability 1307:1315 arg1 design 1356:1361 the design 1352:1361 the design of antigen binding Fc proteins 1352:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 2 71 theme loops 274:278 arg1 Engineering 244:254 Engineering 244:254 Engineering of its structural loops 244:278 Engineering of its structural loops allows generation of an antigen binding site. 24451126 1 72 theme starting 225:232 arg1 scaffold 234:241 starting scaffold 225:241 starting scaffold 225:241 An interesting format in the development of therapeutic monoclonal antibodies uses the crystallizable fragment of IgG1 as starting scaffold. 24451126 5 73 theme fragment 680:687 arg1 fragment 680:687 the Fc fragment 673:687 the Fc fragment of human IgG1 673:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 5 73 theme fragment 680:687 arg1 domains 662:668 the domains 658:668 the domains of the Fc fragment of human IgG1 658:701 In order to understand this cross-talk, molecular dynamics simulations of the domains of the Fc fragment of human IgG1 are reported. 24451126 8 74 theme potential 1120:1128 arg1 sites 1130:1134 potential sites 1120:1134 potential sites for engineering antigen binding sites 1120:1172 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 8 74 theme potential 1120:1128 arg1 Regions 1067:1073 Regions 1067:1073 Regions of high local flexibility 1067:1099 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 2 75 theme structural 263:272 arg1 loops 274:278 its structural loops 259:278 its structural loops 259:278 Engineering of its structural loops allows generation of an antigen binding site. 24451126 9 76 theme IgG1-Fc 1341:1347 arg1 use 1321:1323 use 1321:1323 use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1321:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 9 76 theme IgG1-Fc 1341:1347 arg1 stability 1307:1315 stability 1307:1315 stability 1307:1315 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 8 77 theme high 1078:1081 arg1 flexibility 1089:1099 high local flexibility 1078:1099 high local flexibility 1078:1099 Regions of high local flexibility were identified as potential sites for engineering antigen binding sites. 24451126 9 78 theme approaches 1280:1289 arg1 structure 1239:1247 the available X-ray structure 1219:1247 the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1219:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 6 79 used used 787:790 arg2 point 806:810 a starting point 795:810 a starting point for simulations of the wild-type protein at two different pH values 795:878 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 6 79 used used 787:790 arg2 structure 721:729 The structure 717:729 The structure of human IgG1-Fc obtained from X-ray crystallography 717:782 The structure of human IgG1-Fc obtained from X-ray crystallography is used as a starting point for simulations of the wild-type protein at two different pH values. 24451126 7 80 from domain 942:947 arg1 impact 964:969 the impact 960:969 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 7 80 from domain 942:947 arg1 effect 897:902 The stabilizing effect 881:902 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains 881:1048 The stabilizing effect of a single point mutation in the CH3 domain as well as the impact of the hinge region and the glycan tree structure connected to the CH2 domains is investigated. 24451126 9 81 theme proteins 1385:1392 arg1 design 1356:1361 the design 1352:1361 the design of antigen binding Fc proteins 1352:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 0 82 theme IgG1-insights 64:76 arg1 fragment 52:59 the crystallizable fragment 33:59 the crystallizable fragment of IgG1-insights 33:76 Molecular dynamics simulation of the crystallizable fragment of IgG1-insights for the design of Fcabs. 24451126 9 83 theme X-ray 1233:1237 arg1 structure 1239:1247 the available X-ray structure 1219:1247 the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1219:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 24451126 9 84 theme scaffold 1332:1339 arg1 IgG1-Fc 1341:1347 the scaffold IgG1-Fc 1328:1347 the scaffold IgG1-Fc in the design of antigen binding Fc proteins 1328:1392 Obtained data are discussed with respect to the available X-ray structure of IgG1-Fc, directed evolution approaches that screen for stability and use of the scaffold IgG1-Fc in the design of antigen binding Fc proteins. 25849117 13 0 theme short-lived 1946:1956 arg1 intermediate 1982:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 2 1 theme reaction 357:364 arg1 mechanisms 366:375 the reaction mechanisms 353:375 the reaction mechanisms of the participating enzymes 353:404 To understand the process of protein glycosylation, the reaction mechanisms of the participating enzymes need to be known. 25849117 4 2 theme glycosyltransferase 612:630 arg1 transferase 655:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 13 3 theme oxocarbenium 1969:1980 arg1 intermediate 1982:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 11 4 theme optimized 1662:1670 arg1 higher 1723:1728 higher 1723:1728 higher 1723:1728 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 11 4 theme optimized 1662:1670 arg1 state 1683:1687 The optimized transition state 1658:1687 The optimized transition state for the reaction 1658:1704 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 6 5 theme surface 1221:1227 arg1 knowledge 1187:1195 knowledge 1187:1195 knowledge of the potential energy surface 1187:1227 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 10 6 theme nucleophilic 1604:1615 arg1 substitution 1617:1628 a same-face nucleophilic substitution 1592:1628 a same-face nucleophilic substitution with internal return (SNi) 1592:1655 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 1 7 theme crucial 209:215 arg1 role 217:220 a crucial role 207:220 a crucial role 207:220 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 6 8 theme potential 1204:1212 arg1 surface 1221:1227 the potential energy surface 1200:1227 the potential energy surface 1200:1227 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 11 9 theme product 1782:1788 arg1 complex 1790:1796 the product complex 1778:1796 the product complex 1778:1796 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 0 10 theme glycosyltransferase 126:144 arg1 ppGalNAcT2 146:155 glycosyltransferase ppGalNAcT2 126:155 glycosyltransferase ppGalNAcT2 126:155 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 4 11 theme QM/MM-based 693:703 arg1 surface 743:749 a potential energy surface 724:749 a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 724:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 11 theme QM/MM-based 693:703 arg1 approaches 705:714 coupling two different QM/MM-based approaches 670:714 coupling two different QM/MM-based approaches 670:714 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 1 12 gly glycosylation 162:174 arg1 proteins 192:199 cell surface proteins 179:199 cell surface proteins 179:199 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 4 13 theme difference 772:781 arg1 dimensions 783:792 two distance difference dimensions 759:792 two distance difference dimensions 759:792 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 5 14 theme scan 990:993 arg1 system 1006:1011 a predefined scan coordinate system 977:1011 a predefined scan coordinate system 977:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 5 15 theme predefined 979:988 arg1 system 1006:1011 a predefined scan coordinate system 977:1011 a predefined scan coordinate system 977:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 4 16 theme energy 736:741 arg1 surface 743:749 a potential energy surface 724:749 a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 724:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 16 theme energy 736:741 arg1 approaches 705:714 coupling two different QM/MM-based approaches 670:714 coupling two different QM/MM-based approaches 670:714 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 1 17 theme cell 270:273 arg1 adhesion 275:282 cell adhesion 270:282 cell adhesion 270:282 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 13 18 theme high-level 2066:2075 arg1 functionals 2085:2095 high-level density functionals 2066:2095 high-level density functionals 2066:2095 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 5 19 theme reactive 951:958 arg1 processes 960:968 reactive processes 951:968 reactive processes due to a predefined scan coordinate system 951:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 8 20 theme structural 1380:1389 arg1 model 1391:1395 The structural model 1376:1395 The structural model 1376:1395 The structural model is based on the crystal structure of human isoform 2. 25849117 5 21 theme due 970:972 arg1 processes 960:968 reactive processes 951:968 reactive processes due to a predefined scan coordinate system 951:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 4 22 theme reaction 815:822 arg1 optimisation 829:840 a minimum energy reaction path optimisation 798:840 a minimum energy reaction path optimisation 798:840 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 0 23 theme PES 96:98 arg1 calculations 100:111 PES calculations 96:111 PES calculations 96:111 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 13 24 theme energy 2035:2040 arg1 profiles 2042:2049 the reaction energy profiles 2022:2049 the reaction energy profiles obtained using high-level density functionals 2022:2095 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 4 25 dep surface 743:749 arg1 scan 751:754 scan 751:754 scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 751:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 6 26 theme path 1032:1035 arg1 methods 1050:1056 path optimisation methods 1032:1056 path optimisation methods 1032:1056 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 3 27 theme reaction 437:444 arg1 mechanism 446:454 the reaction mechanism 433:454 the reaction mechanism of retaining glycosyltransferases 433:488 However, the reaction mechanism of retaining glycosyltransferases has not yet been sufficiently explained. 25849117 8 28 theme crystal 1413:1419 arg1 structure 1421:1429 the crystal structure 1409:1429 the crystal structure of human isoform 2 1409:1448 The structural model is based on the crystal structure of human isoform 2. 25849117 2 29 theme enzymes 398:404 arg1 mechanisms 366:375 the reaction mechanisms 353:375 the reaction mechanisms of the participating enzymes 353:404 To understand the process of protein glycosylation, the reaction mechanisms of the participating enzymes need to be known. 25849117 0 30 theme Stepwise 0:7 arg1 mechanism 19:27 Stepwise catalytic mechanism 0:27 Stepwise catalytic mechanism via short-lived intermediate 0:56 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 6 31 theme dimensions 1113:1122 arg1 number 1103:1108 a virtually unlimited number 1081:1108 a virtually unlimited number of dimensions 1081:1122 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 4 32 theme minimum 800:806 arg1 optimisation 829:840 a minimum energy reaction path optimisation 798:840 a minimum energy reaction path optimisation 798:840 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 3 33 theme glycosyltransferases 469:488 arg1 mechanism 446:454 the reaction mechanism 433:454 the reaction mechanism of retaining glycosyltransferases 433:488 However, the reaction mechanism of retaining glycosyltransferases has not yet been sufficiently explained. 25849117 5 34 theme Potential 880:888 arg1 studies 902:908 Potential energy scan studies 880:908 Potential energy scan studies 880:908 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 12 35 theme attack 1856:1861 arg1 process 1832:1838 the process 1828:1838 the process of nucleophilic attack 1828:1861 During the process of nucleophilic attack, a proton is synchronously transferred to the leaving phosphate. 25849117 6 36 theme number 1103:1108 arg1 sampling 1069:1076 the sampling 1065:1076 the sampling of a virtually unlimited number of dimensions 1065:1122 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 4 37 theme isoform 585:591 arg1 mechanism 566:574 the catalytic mechanism 552:574 the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 552:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 8 38 theme isoform 1440:1446 arg1 structure 1421:1429 the crystal structure 1409:1429 the crystal structure of human isoform 2 1409:1448 The structural model is based on the crystal structure of human isoform 2. 25849117 0 39 theme short-lived 33:43 arg1 intermediate 45:56 short-lived intermediate 33:56 short-lived intermediate 33:56 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 0 40 theme combined 72:79 arg1 MERP 87:90 combined QM/MM MERP 72:90 combined QM/MM MERP 72:90 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 10 41 with substitution 1617:1628 arg1 SNi 1652:1654 SNi 1652:1654 SNi 1652:1654 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 10 41 with substitution 1617:1628 arg1 return 1644:1649 internal return 1635:1649 internal return (SNi) 1635:1655 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 7 42 theme inherent 1338:1345 arg1 errors 1331:1336 potential errors 1321:1336 potential errors inherent to each of these approaches 1321:1373 By combining these methods, we have been able to eliminate the most significant sources of potential errors inherent to each of these approaches. 25849117 4 43 theme catalytic 556:564 arg1 mechanism 566:574 the catalytic mechanism 552:574 the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 552:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 5 44 dep studies 902:908 arg1 scan 897:900 scan 897:900 scan 897:900 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 8 45 theme human 1434:1438 arg1 isoform 1440:1446 human isoform 2 1434:1448 human isoform 2 1434:1448 The structural model is based on the crystal structure of human isoform 2. 25849117 2 46 theme glycosylation 338:350 arg1 process 319:325 the process 315:325 the process of protein glycosylation 315:350 To understand the process of protein glycosylation, the reaction mechanisms of the participating enzymes need to be known. 25849117 7 47 theme potential 1321:1329 arg1 errors 1331:1336 potential errors 1321:1336 potential errors inherent to each of these approaches 1321:1373 By combining these methods, we have been able to eliminate the most significant sources of potential errors inherent to each of these approaches. 25849117 4 48 theme polypeptide 632:642 arg1 transferase 655:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 49 theme Elastic 859:865 arg1 method 872:877 the Nudged Elastic Band method 848:877 the Nudged Elastic Band method 848:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 1 50 theme processes 251:259 arg1 multitude 227:235 a multitude 225:235 a multitude of biological processes, such as cell adhesion and recognition 225:298 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 4 51 theme transferase 655:665 arg1 isoform 585:591 human isoform 2 579:593 human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 579:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 13 52 theme metastable 1958:1967 arg1 intermediate 1982:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 a short-lived metastable oxocarbenium intermediate 1944:1993 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 9 53 theme QM/MM 1458:1462 arg1 method 1464:1469 the QM/MM method 1454:1469 the QM/MM method 1454:1469 In the QM/MM method, the QM region consists of 275 atoms, the remaining 5776 atoms were in the MM region. 25849117 5 54 theme processes 960:968 arg1 sampling 939:946 inadequate sampling 928:946 inadequate sampling of reactive processes due to a predefined scan coordinate system 928:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 4 55 theme retaining 602:610 arg1 transferase 655:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 13 56 theme intermediate 1982:1993 arg1 presence 1932:1939 The presence 1928:1939 The presence of a short-lived metastable oxocarbenium intermediate 1928:1993 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 13 56 theme intermediate 1982:1993 arg1 likely 1998:2003 likely 1998:2003 likely 1998:2003 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 9 57 theme QM 1476:1477 arg1 region 1479:1484 the QM region 1472:1484 the QM region 1472:1484 In the QM/MM method, the QM region consists of 275 atoms, the remaining 5776 atoms were in the MM region. 25849117 1 58 theme proteins 192:199 arg1 glycosylation 162:174 The glycosylation 158:174 The glycosylation of cell surface proteins 158:199 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 11 59 theme transition 1672:1681 arg1 higher 1723:1728 higher 1723:1728 higher 1723:1728 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 11 59 theme transition 1672:1681 arg1 state 1683:1687 The optimized transition state 1658:1687 The optimized transition state for the reaction 1658:1704 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 5 60 theme inadequate 928:937 arg1 sampling 939:946 inadequate sampling 928:946 inadequate sampling of reactive processes due to a predefined scan coordinate system 928:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 6 61 theme energy 1214:1219 arg1 surface 1221:1227 the potential energy surface 1200:1227 the potential energy surface 1200:1227 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 10 62 theme same-face 1594:1602 arg1 substitution 1617:1628 a same-face nucleophilic substitution 1592:1628 a same-face nucleophilic substitution with internal return (SNi) 1592:1655 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 5 63 theme energy 890:895 arg1 studies 902:908 Potential energy scan studies 880:908 Potential energy scan studies 880:908 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 6 64 theme same 1021:1024 arg1 time 1026:1029 the same time 1017:1029 the same time 1017:1029 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 1 65 theme surface 184:190 arg1 proteins 192:199 cell surface proteins 179:199 cell surface proteins 179:199 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 4 66 theme coupling 670:677 arg1 surface 743:749 a potential energy surface 724:749 a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 724:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 66 theme coupling 670:677 arg1 approaches 705:714 coupling two different QM/MM-based approaches 670:714 coupling two different QM/MM-based approaches 670:714 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 11 67 theme complex 1790:1796 arg1 energy 1768:1773 the energy 1764:1773 the energy of the product complex 1764:1796 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 11 67 theme complex 1790:1796 arg1 lower 1814:1818 lower 1814:1818 lower 1814:1818 The optimized transition state for the reaction is 13.8 kcal/mol higher in energy than the reactant while the energy of the product complex is 6.7 kcal/mol lower. 25849117 4 68 theme different 683:691 arg1 surface 743:749 a potential energy surface 724:749 a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 724:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 68 theme different 683:691 arg1 approaches 705:714 coupling two different QM/MM-based approaches 670:714 coupling two different QM/MM-based approaches 670:714 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 10 69 theme internal 1635:1642 arg1 SNi 1652:1654 SNi 1652:1654 SNi 1652:1654 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 10 69 theme internal 1635:1642 arg1 return 1644:1649 internal return 1635:1649 internal return (SNi) 1635:1655 We found that ppGalNAcT2 catalyzes a same-face nucleophilic substitution with internal return (SNi). 25849117 9 70 theme remaining 1513:1521 arg1 atoms 1528:1532 the remaining 5776 atoms 1509:1532 the remaining 5776 atoms 1509:1532 In the QM/MM method, the QM region consists of 275 atoms, the remaining 5776 atoms were in the MM region. 25849117 13 71 theme reaction 2026:2033 arg1 profiles 2042:2049 the reaction energy profiles 2022:2049 the reaction energy profiles obtained using high-level density functionals 2022:2095 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 4 72 theme distance 763:770 arg1 dimensions 783:792 two distance difference dimensions 759:792 two distance difference dimensions 759:792 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 0 73 theme QM/MM 81:85 arg1 MERP 87:90 combined QM/MM MERP 72:90 combined QM/MM MERP 72:90 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 4 74 theme potential 726:734 arg1 surface 743:749 a potential energy surface 724:749 a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method 724:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 74 theme potential 726:734 arg1 approaches 705:714 coupling two different QM/MM-based approaches 670:714 coupling two different QM/MM-based approaches 670:714 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 13 75 theme density 2077:2083 arg1 functionals 2085:2095 high-level density functionals 2066:2095 high-level density functionals 2066:2095 The presence of a short-lived metastable oxocarbenium intermediate is likely, as indicated by the reaction energy profiles obtained using high-level density functionals. 25849117 12 76 theme leaving 1909:1915 arg1 phosphate 1917:1925 the leaving phosphate 1905:1925 the leaving phosphate 1905:1925 During the process of nucleophilic attack, a proton is synchronously transferred to the leaving phosphate. 25849117 1 77 theme cell 179:182 arg1 proteins 192:199 cell surface proteins 179:199 cell surface proteins 179:199 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 9 78 theme MM 1546:1547 arg1 region 1549:1554 the MM region 1542:1554 the MM region 1542:1554 In the QM/MM method, the QM region consists of 275 atoms, the remaining 5776 atoms were in the MM region. 25849117 6 79 theme optimisation 1037:1048 arg1 methods 1050:1056 path optimisation methods 1032:1056 path optimisation methods 1032:1056 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 4 80 theme energy 808:813 arg1 optimisation 829:840 a minimum energy reaction path optimisation 798:840 a minimum energy reaction path optimisation 798:840 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 4 81 theme path 824:827 arg1 optimisation 829:840 a minimum energy reaction path optimisation 798:840 a minimum energy reaction path optimisation 798:840 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 0 82 theme catalytic 9:17 arg1 mechanism 19:27 Stepwise catalytic mechanism 0:27 Stepwise catalytic mechanism via short-lived intermediate 0:56 Stepwise catalytic mechanism via short-lived intermediate inferred from combined QM/MM MERP and PES calculations on retaining glycosyltransferase ppGalNAcT2. 25849117 5 83 theme coordinate 995:1004 arg1 system 1006:1011 a predefined scan coordinate system 977:1011 a predefined scan coordinate system 977:1011 Potential energy scan studies often suffer from inadequate sampling of reactive processes due to a predefined scan coordinate system. 25849117 4 84 theme human 579:583 arg1 isoform 585:591 human isoform 2 579:593 human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 579:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 2 85 theme participating 384:396 arg1 enzymes 398:404 the participating enzymes 380:404 the participating enzymes 380:404 To understand the process of protein glycosylation, the reaction mechanisms of the participating enzymes need to be known. 25849117 6 86 theme unlimited 1093:1101 arg1 number 1103:1108 a virtually unlimited number 1081:1108 a virtually unlimited number of dimensions 1081:1122 At the same time, path optimisation methods enable the sampling of a virtually unlimited number of dimensions, but their results cannot be unambiguously interpreted without knowledge of the potential energy surface. 25849117 4 87 theme Band 867:870 arg1 method 872:877 the Nudged Elastic Band method 848:877 the Nudged Elastic Band method 848:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 3 88 theme retaining 459:467 arg1 glycosyltransferases 469:488 retaining glycosyltransferases 459:488 retaining glycosyltransferases 459:488 However, the reaction mechanism of retaining glycosyltransferases has not yet been sufficiently explained. 25849117 7 89 theme errors 1331:1336 arg1 sources 1310:1316 the most significant sources 1289:1316 the most significant sources of potential errors inherent to each of these approaches 1289:1373 By combining these methods, we have been able to eliminate the most significant sources of potential errors inherent to each of these approaches. 25849117 12 90 theme nucleophilic 1843:1854 arg1 attack 1856:1861 nucleophilic attack 1843:1861 nucleophilic attack 1843:1861 During the process of nucleophilic attack, a proton is synchronously transferred to the leaving phosphate. 25849117 4 91 theme UDP-GalNAc 644:653 arg1 transferase 655:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 the retaining glycosyltransferase polypeptide UDP-GalNAc transferase 598:665 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 25849117 9 92 dep atoms 1502:1506 arg1 region 1549:1554 the MM region 1542:1554 the MM region 1542:1554 In the QM/MM method, the QM region consists of 275 atoms, the remaining 5776 atoms were in the MM region. 25849117 2 93 theme protein 330:336 arg1 glycosylation 338:350 protein glycosylation 330:350 protein glycosylation 330:350 To understand the process of protein glycosylation, the reaction mechanisms of the participating enzymes need to be known. 25849117 1 94 theme biological 240:249 arg1 processes 251:259 biological processes 240:259 biological processes 240:259 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 1 94 theme biological 240:249 arg1 recognition 288:298 recognition 288:298 recognition 288:298 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 1 94 theme biological 240:249 arg1 adhesion 275:282 cell adhesion 270:282 cell adhesion 270:282 The glycosylation of cell surface proteins plays a crucial role in a multitude of biological processes, such as cell adhesion and recognition. 25849117 7 95 theme significant 1298:1308 arg1 sources 1310:1316 the most significant sources 1289:1316 the most significant sources of potential errors inherent to each of these approaches 1289:1373 By combining these methods, we have been able to eliminate the most significant sources of potential errors inherent to each of these approaches. 25849117 4 96 theme Nudged 852:857 arg1 method 872:877 the Nudged Elastic Band method 848:877 the Nudged Elastic Band method 848:877 Here we investigated the catalytic mechanism of human isoform 2 of the retaining glycosyltransferase polypeptide UDP-GalNAc transferase by coupling two different QM/MM-based approaches, namely a potential energy surface scan in two distance difference dimensions and a minimum energy reaction path optimisation using the Nudged Elastic Band method. 26388028 2 0 theme X-ray 389:393 arg1 studies 367:373 structural studies 356:373 structural studies 356:373 This sample heterogeneity can be problematic for structural studies, particularly X-ray crystallography. 26388028 2 0 theme X-ray 389:393 arg1 crystallography 395:409 X-ray crystallography 389:409 X-ray crystallography 389:409 This sample heterogeneity can be problematic for structural studies, particularly X-ray crystallography. 26388028 1 1 theme large 270:274 arg1 number 276:281 a large number 268:281 a large number of possible glycoforms 268:304 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 0 2 theme Protein 67:73 arg1 Refinement 19:28 Refinement 19:28 Refinement 19:28 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 0 2 theme Protein 67:73 arg1 Building 6:13 Model Building 0:13 Model Building 0:13 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 3 3 gly glycosylated 448:459 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 3 3 gly glycosylated 448:459 arg1 proteins 461:468 heavily glycosylated proteins 440:468 heavily glycosylated proteins such as the HIV-1 Env viral spike protein 440:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 1 4 from cells 166:170 arg1 proteins 142:149 Secretory and membrane proteins 119:149 proteins 142:149 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 0 5 theme Cryoelectron 94:105 arg1 Microscopy 107:116 High-Resolution Cryoelectron Microscopy 78:116 High-Resolution Cryoelectron Microscopy 78:116 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 1 6 link N-linked 224:231 arg1 glycosylation 233:245 N-linked glycosylation 224:245 N-linked glycosylation 224:245 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 2 7 theme sample 312:317 arg1 heterogeneity 319:331 This sample heterogeneity 307:331 This sample heterogeneity 307:331 This sample heterogeneity can be problematic for structural studies, particularly X-ray crystallography. 26388028 0 8 theme High-Resolution 78:92 arg1 Microscopy 107:116 High-Resolution Cryoelectron Microscopy 78:116 High-Resolution Cryoelectron Microscopy 78:116 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 5 9 theme Env 925:927 arg1 structures 929:938 natively glycosylated HIV-1 Env structures 897:938 natively glycosylated HIV-1 Env structures 897:938 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 1 10 theme possible 286:293 arg1 glycoforms 295:304 possible glycoforms 286:304 possible glycoforms 286:304 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 3 11 theme glycans 561:567 arg1 majority 549:556 the majority 545:556 the majority of glycans 545:567 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 1 12 theme post-translational 180:197 arg1 glycosylation 233:245 N-linked glycosylation 224:245 N-linked glycosylation 224:245 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 1 12 theme post-translational 180:197 arg1 modifications 199:211 post-translational modifications 180:211 post-translational modifications 180:211 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 4 13 theme Endoglycosidase 617:631 arg1 EndoH 636:640 EndoH 636:640 EndoH 636:640 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 4 13 theme Endoglycosidase 617:631 arg1 H 633:633 Endoglycosidase H 617:633 Endoglycosidase H (EndoH) 617:641 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 5 14 theme single-particle 797:811 arg1 microscopy 826:835 single-particle cryoelectron microscopy 797:835 single-particle cryoelectron microscopy 797:835 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 3 15 theme HIV-1 482:486 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 3 16 theme crystal 418:424 arg1 structures 426:435 crystal structures 418:435 crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein 418:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 5 17 theme Å 960:960 arg1 resolution 962:971 4.36 Å resolution 955:971 4.36 Å resolution 955:971 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 0 18 theme Model 0:4 arg1 Building 6:13 Model Building 0:13 Model Building 0:13 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 1 19 theme glycoforms 295:304 arg1 number 276:281 a large number 268:281 a large number of possible glycoforms 268:304 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 4 20 theme native 735:740 arg1 glycoform 742:750 the native glycoform 731:750 the native glycoform 731:750 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 4 20 theme native 735:740 arg1 form 706:709 the high-mannose form 689:709 the high-mannose form 689:709 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 6 21 theme complete 1016:1023 arg1 epitope 1025:1031 the complete epitope 1012:1031 the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans 1012:1141 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 22 from epitope 1025:1031 arg1 context 1091:1097 the context 1087:1097 the context of the trimer expressed with native glycans 1087:1141 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 23 theme antibody 1059:1066 arg1 epitope 1025:1031 the complete epitope 1012:1031 the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans 1012:1141 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 1 24 theme N-linked 224:231 arg1 glycosylation 233:245 N-linked glycosylation 224:245 N-linked glycosylation 224:245 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 6 25 theme native 1128:1133 arg1 glycans 1135:1141 native glycans 1128:1141 native glycans 1128:1141 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 5 26 theme glycosylated 906:917 arg1 structures 929:938 natively glycosylated HIV-1 Env structures 897:938 natively glycosylated HIV-1 Env structures 897:938 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 3 27 theme glycosylated 448:459 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 3 27 theme glycosylated 448:459 arg1 proteins 461:468 heavily glycosylated proteins 440:468 heavily glycosylated proteins such as the HIV-1 Env viral spike protein 440:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 4 28 theme expressed 665:673 arg1 glycans 675:681 all expressed glycans 661:681 all expressed glycans 661:681 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 3 29 theme Env 488:490 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 3 30 theme proteins 461:468 arg1 structures 426:435 crystal structures 418:435 crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein 418:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 5 31 theme cryoelectron 813:824 arg1 microscopy 826:835 single-particle cryoelectron microscopy 797:835 single-particle cryoelectron microscopy 797:835 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 3 32 theme viral 492:496 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 0 33 theme Glycosylated 44:55 arg1 Protein 67:73 a Natively Glycosylated HIV-1 Env Protein 33:73 a Natively Glycosylated HIV-1 Env Protein 33:73 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 5 34 theme improved 772:779 arg1 technologies 781:792 significantly improved technologies 758:792 significantly improved technologies in single-particle cryoelectron microscopy 758:835 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 6 35 theme trimer 1106:1111 arg1 context 1091:1097 the context 1087:1097 the context of the trimer expressed with native glycans 1087:1141 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 1 36 theme Secretory 119:127 arg1 proteins 142:149 Secretory and membrane proteins 119:149 proteins 142:149 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 5 37 gly glycosylated 906:917 arg1 structures 929:938 natively glycosylated HIV-1 Env structures 897:938 natively glycosylated HIV-1 Env structures 897:938 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 5 38 from technologies 781:792 arg1 microscopy 826:835 single-particle cryoelectron microscopy 797:835 single-particle cryoelectron microscopy 797:835 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 1 39 theme membrane 133:140 arg1 proteins 142:149 Secretory and membrane proteins 119:149 proteins 142:149 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26388028 6 40 theme broadly 1038:1044 arg1 bnAb 1069:1072 bnAb 1069:1072 bnAb 1069:1072 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 40 theme broadly 1038:1044 arg1 PGT128 1076:1081 PGT128 1076:1081 PGT128 1076:1081 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 40 theme broadly 1038:1044 arg1 antibody 1059:1066 a broadly neutralizing antibody 1036:1066 a broadly neutralizing antibody (bnAb) 1036:1073 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 4 41 theme high-mannose 693:704 arg1 form 706:709 the high-mannose form 689:709 the high-mannose form 689:709 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 4 41 theme high-mannose 693:704 arg1 glycoform 742:750 the native glycoform 731:750 the native glycoform 731:750 This step is most frequently carried out using Endoglycosidase H (EndoH) and requires that all expressed glycans be in the high-mannose form, which is often not the native glycoform. 26388028 2 42 theme structural 356:365 arg1 studies 367:373 structural studies 356:373 structural studies 356:373 This sample heterogeneity can be problematic for structural studies, particularly X-ray crystallography. 26388028 2 42 theme structural 356:365 arg1 crystallography 395:409 X-ray crystallography 389:409 X-ray crystallography 389:409 This sample heterogeneity can be problematic for structural studies, particularly X-ray crystallography. 26388028 0 43 theme Env 63:65 arg1 Protein 67:73 a Natively Glycosylated HIV-1 Env Protein 33:73 a Natively Glycosylated HIV-1 Env Protein 33:73 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 6 44 theme neutralizing 1046:1057 arg1 bnAb 1069:1072 bnAb 1069:1072 bnAb 1069:1072 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 44 theme neutralizing 1046:1057 arg1 PGT128 1076:1081 PGT128 1076:1081 PGT128 1076:1081 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 6 44 theme neutralizing 1046:1057 arg1 antibody 1059:1066 a broadly neutralizing antibody 1036:1066 a broadly neutralizing antibody (bnAb) 1036:1073 At this resolution we can now analyze the complete epitope of a broadly neutralizing antibody (bnAb), PGT128, in the context of the trimer expressed with native glycans. 26388028 3 45 theme spike 498:502 arg1 protein 504:510 the HIV-1 Env viral spike protein 478:510 the HIV-1 Env viral spike protein 478:510 Thus, crystal structures of heavily glycosylated proteins such as the HIV-1 Env viral spike protein have been determined by removing the majority of glycans. 26388028 5 46 theme HIV-1 919:923 arg1 structures 929:938 natively glycosylated HIV-1 Env structures 897:938 natively glycosylated HIV-1 Env structures 897:938 With significantly improved technologies in single-particle cryoelectron microscopy, we demonstrate that it is now possible to refine and build natively glycosylated HIV-1 Env structures in solution to 4.36 Å resolution. 26388028 0 47 theme HIV-1 57:61 arg1 Protein 67:73 a Natively Glycosylated HIV-1 Env Protein 33:73 a Natively Glycosylated HIV-1 Env Protein 33:73 Model Building and Refinement of a Natively Glycosylated HIV-1 Env Protein by High-Resolution Cryoelectron Microscopy. 26388028 1 48 theme mammalian 156:164 arg1 cells 166:170 mammalian cells 156:170 mammalian cells 156:170 Secretory and membrane proteins from mammalian cells undergo post-translational modifications, including N-linked glycosylation, which can result in a large number of possible glycoforms. 26328495 5 0 mod modified 758:765 arg3 O-glycan 790:797 a sialylated core-1 O-glycan 770:797 a sialylated core-1 O-glycan 770:797 These results suggest that POMGNT1 is modified by a sialylated core-1 O-glycan. 26328495 5 0 mod modified 758:765 arg1 POMGNT1 747:753 POMGNT1 747:753 POMGNT1 747:753 These results suggest that POMGNT1 is modified by a sialylated core-1 O-glycan. 26328495 7 1 theme NeuAc 1004:1008 arg1 HexNAc 1018:1023 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 4 2 theme recombinant 525:535 arg1 POMGNT1 537:543 The recombinant POMGNT1 521:543 The recombinant POMGNT1 521:543 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 3 3 from cells 514:518 arg1 POMGNT1 493:499 recombinant human POMGNT1 475:499 recombinant human POMGNT1 from HEK293T cells 475:518 To determine whether POMGNT1 is O-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. 26328495 4 4 theme lectin 706:711 arg1 reactivity 675:684 the reactivity 671:684 the reactivity of Arachis hypogaea lectin (PNA) 671:717 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 1 5 theme O-linked 57:64 arg1 β1,2-N-acetylglucosaminyltransferase 74:109 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 49:111 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 49:121 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 5 theme O-linked 57:64 arg1 POMGNT1 114:120 POMGNT1 114:120 POMGNT1 114:120 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 5 theme O-linked 57:64 arg1 glycosyltransferase 134:152 a Golgi glycosyltransferase 126:152 a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 126:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 7 6 dep HexNAc 1018:1023 arg1 2 1079:1079 2 1079:1079 2 1079:1079 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 7 theme several 963:969 arg1 O-glycans 982:990 several mucin-type O-glycans 963:990 several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2 963:1079 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 1 8 theme mannose 66:72 arg1 β1,2-N-acetylglucosaminyltransferase 74:109 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 49:111 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 49:121 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 8 theme mannose 66:72 arg1 POMGNT1 114:120 POMGNT1 114:120 POMGNT1 114:120 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 8 theme mannose 66:72 arg1 glycosyltransferase 134:152 a Golgi glycosyltransferase 126:152 a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 126:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 9 link O-linked 57:64 arg1 β1,2-N-acetylglucosaminyltransferase 74:109 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 49:111 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 49:121 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 9 link O-linked 57:64 arg1 POMGNT1 114:120 POMGNT1 114:120 POMGNT1 114:120 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 9 link O-linked 57:64 arg1 glycosyltransferase 134:152 a Golgi glycosyltransferase 126:152 a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 126:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 6 10 theme pyrazolone-labeling 882:900 arg1 methods 902:908 β-elimination and pyrazolone-labeling methods 864:908 β-elimination and pyrazolone-labeling methods 864:908 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 3 11 theme recombinant 475:485 arg1 POMGNT1 493:499 recombinant human POMGNT1 475:499 recombinant human POMGNT1 from HEK293T cells 475:518 To determine whether POMGNT1 is O-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. 26328495 6 12 from structures 822:831 arg1 POMGNT1 853:859 POMGNT1 853:859 POMGNT1 853:859 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 4 13 theme sialidase 602:610 arg1 digestion 612:620 sialidase digestion 602:620 sialidase digestion of POMGNT1 602:631 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 2 14 mod modified 392:399 arg1 POMGNT1 381:387 POMGNT1 381:387 POMGNT1 381:387 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 2 14 mod modified 392:399 arg3 O-glycans 404:412 O-glycans 404:412 O-glycans 404:412 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 1 15 theme N-acetylglucosamine 190:208 arg1 linkage 227:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage 186:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 186:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 8 16 theme POMGNT1 1154:1160 arg1 properties 1140:1149 properties 1140:1149 properties 1140:1149 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 16 theme POMGNT1 1154:1160 arg1 functions 1126:1134 functions 1126:1134 functions 1126:1134 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 6 17 theme β-elimination 864:876 arg1 methods 902:908 β-elimination and pyrazolone-labeling methods 864:908 β-elimination and pyrazolone-labeling methods 864:908 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 8 18 theme sPOMGNT1 1230:1237 arg1 forms 1209:1213 non-glycosylated forms 1192:1213 non-glycosylated forms of recombinant sPOMGNT1 for their activity 1192:1256 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 18 theme sPOMGNT1 1230:1237 arg1 hydrophobicity 1270:1283 surface hydrophobicity 1262:1283 surface hydrophobicity 1262:1283 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 9 19 theme surface 1375:1381 arg1 hydrophobicity 1383:1396 surface hydrophobicity 1375:1396 surface hydrophobicity 1375:1396 POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of O-glycans. 26328495 6 20 from O-glycans 840:848 arg1 POMGNT1 853:859 POMGNT1 853:859 POMGNT1 853:859 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 4 21 theme sieboldiana 572:582 arg1 SSA 592:594 SSA 592:594 SSA 592:594 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 4 21 theme sieboldiana 572:582 arg1 lectin 584:589 Sambucus sieboldiana lectin 563:589 Sambucus sieboldiana lectin (SSA) 563:595 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 2 22 theme N-glycosylation 325:339 arg1 sites 341:345 no potential N-glycosylation sites 312:345 no potential N-glycosylation sites 312:345 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 7 23 theme NeuAc 1058:1062 arg1 Hex 1066:1068 (NeuAc)2(Hex)2 1057:1070 (NeuAc)2(Hex)2(HexNAc) 1057:1078 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 23 theme NeuAc 1058:1062 arg1 HexNAc 1072:1077 HexNAc 1072:1077 HexNAc 1072:1077 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 4 24 theme Sambucus 563:570 arg1 SSA 592:594 SSA 592:594 SSA 592:594 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 4 24 theme Sambucus 563:570 arg1 lectin 584:589 Sambucus sieboldiana lectin 563:589 Sambucus sieboldiana lectin (SSA) 563:595 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 7 25 theme mucin-type 971:980 arg1 O-glycans 982:990 several mucin-type O-glycans 963:990 several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2 963:1079 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 2 26 theme potential 315:323 arg1 sites 341:345 no potential N-glycosylation sites 312:345 no potential N-glycosylation sites 312:345 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 4 27 theme SSA 643:645 arg1 reactivity 647:656 SSA reactivity 643:656 SSA reactivity 643:656 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 6 28 from POMGNT1 853:859 arg1 structures 822:831 the structures 818:831 the structures of the O-glycans on POMGNT1 818:859 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 8 29 theme probe 1307:1311 arg1 ANS 1348:1350 ANS 1348:1350 ANS 1348:1350 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 29 theme probe 1307:1311 arg1 sulfonate 1337:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate 1291:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS) 1291:1351 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 1 30 theme β1→2Man 219:225 arg1 linkage 227:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage 186:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 186:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 5 31 theme sialylated 772:781 arg1 O-glycan 790:797 a sialylated core-1 O-glycan 770:797 a sialylated core-1 O-glycan 770:797 These results suggest that POMGNT1 is modified by a sialylated core-1 O-glycan. 26328495 3 32 theme HEK293T 506:512 arg1 cells 514:518 HEK293T cells 506:518 HEK293T cells 506:518 To determine whether POMGNT1 is O-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. 26328495 8 33 theme 1-anilino-8-naphthalene 1313:1335 arg1 ANS 1348:1350 ANS 1348:1350 ANS 1348:1350 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 33 theme 1-anilino-8-naphthalene 1313:1335 arg1 sulfonate 1337:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate 1291:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS) 1291:1351 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 9 34 dep presence 1423:1430 arg1 the 1419:1421 the 1419:1421 the 1419:1421 POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of O-glycans. 26328495 1 35 theme linkage 227:233 arg1 formation 173:181 the formation 169:181 the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 169:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 0 36 theme Mucin-Type 27:36 arg1 O-Glycans 38:46 Mucin-Type O-Glycans 27:46 Mucin-Type O-Glycans 27:46 POMGNT1 Is Glycosylated by Mucin-Type O-Glycans. 26328495 5 37 theme core-1 783:788 arg1 O-glycan 790:797 a sialylated core-1 O-glycan 770:797 a sialylated core-1 O-glycan 770:797 These results suggest that POMGNT1 is modified by a sialylated core-1 O-glycan. 26328495 8 38 dep compared 1166:1173 arg1 glycosylated 1175:1186 glycosylated 1175:1186 compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS) 1166:1351 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 7 39 contain containing 992:1001 arg2 2 1035:1035 2 1035:1035 2 1035:1035 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg2 Hex 1037:1039 Hex 1037:1039 Hex 1037:1039 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg2 NeuAc 1029:1033 NeuAc 1029:1033 NeuAc 1029:1033 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg1 O-glycans 982:990 several mucin-type O-glycans 963:990 several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2 963:1079 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg2 Hex 1066:1068 (NeuAc)2(Hex)2 1057:1070 (NeuAc)2(Hex)2(HexNAc) 1057:1078 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg2 HexNAc 1018:1023 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 7 39 contain containing 992:1001 arg2 HexNAc 1072:1077 HexNAc 1072:1077 HexNAc 1072:1077 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 9 40 theme O-glycans 1446:1454 arg1 absence 1435:1441 absence 1435:1441 absence 1435:1441 POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of O-glycans. 26328495 9 40 theme O-glycans 1446:1454 arg1 presence 1423:1430 presence 1423:1430 presence 1423:1430 POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of O-glycans. 26328495 8 41 theme non-glycosylated 1192:1207 arg1 forms 1209:1213 non-glycosylated forms 1192:1213 non-glycosylated forms of recombinant sPOMGNT1 for their activity 1192:1256 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 6 42 theme O-glycans 840:848 arg1 structures 822:831 the structures 818:831 the structures of the O-glycans on POMGNT1 818:859 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 9 43 theme POMGNT1 1354:1360 arg1 activity 1362:1369 POMGNT1 activity 1354:1369 POMGNT1 activity 1354:1369 POMGNT1 activity and surface hydrophobicity were not affected by the presence or absence of O-glycans. 26328495 8 44 theme recombinant 1218:1228 arg1 sPOMGNT1 1230:1237 recombinant sPOMGNT1 1218:1237 recombinant sPOMGNT1 for their activity 1218:1256 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 6 45 with combination 913:923 arg1 spectrometry 935:946 mass spectrometry 930:946 mass spectrometry 930:946 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 8 46 gly non-glycosylated 1192:1207 arg1 forms 1209:1213 non-glycosylated forms 1192:1213 non-glycosylated forms of recombinant sPOMGNT1 for their activity 1192:1256 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 46 gly non-glycosylated 1192:1207 arg1 sPOMGNT1 1230:1237 recombinant sPOMGNT1 1218:1237 recombinant sPOMGNT1 for their activity 1218:1256 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 1 47 theme Golgi 128:132 arg1 glycosyltransferase 134:152 a Golgi glycosyltransferase 126:152 a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 126:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 47 theme Golgi 128:132 arg1 β1,2-N-acetylglucosaminyltransferase 74:109 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 49:111 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 49:121 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 8 48 theme hydrophobic 1295:1305 arg1 ANS 1348:1350 ANS 1348:1350 ANS 1348:1350 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 8 48 theme hydrophobic 1295:1305 arg1 sulfonate 1337:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate 1291:1345 the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS) 1291:1351 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 3 49 gly O-glycosylated 447:460 arg1 POMGNT1 436:442 POMGNT1 436:442 POMGNT1 436:442 To determine whether POMGNT1 is O-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. 26328495 7 50 theme Hex 1012:1014 arg1 HexNAc 1018:1023 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 (NeuAc)1(Hex)1(HexNAc)1 1003:1025 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 3 51 theme human 487:491 arg1 POMGNT1 493:499 recombinant human POMGNT1 475:499 recombinant human POMGNT1 from HEK293T cells 475:518 To determine whether POMGNT1 is O-glycosylated, we prepared recombinant human POMGNT1 from HEK293T cells. 26328495 1 52 theme O-mannosyl 238:247 arg1 glycan 249:254 O-mannosyl glycan 238:254 O-mannosyl glycan 238:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 5 53 gly sialylated 772:781 arg1 O-glycan 790:797 a sialylated core-1 O-glycan 770:797 a sialylated core-1 O-glycan 770:797 These results suggest that POMGNT1 is modified by a sialylated core-1 O-glycan. 26328495 8 54 dep functions 1126:1134 arg1 the 1122:1124 the 1122:1124 the 1122:1124 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 26328495 1 55 theme glycan 249:254 arg1 linkage 227:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage 186:233 the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 186:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 2 56 gly N-glycosylation 325:339 arg2 sites 341:345 no potential N-glycosylation sites 312:345 no potential N-glycosylation sites 312:345 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 7 57 dep 2 1035:1035 arg1 HexNAc 1043:1048 1(HexNAc)1 1041:1050 (NeuAc)2(Hex)1(HexNAc)1 1028:1050 We identified several mucin-type O-glycans containing (NeuAc)1(Hex)1(HexNAc)1, (NeuAc)2(Hex)1(HexNAc)1, and (NeuAc)2(Hex)2(HexNAc)2. 26328495 4 58 theme hypogaea 697:704 arg1 lectin 706:711 Arachis hypogaea lectin 689:711 Arachis hypogaea lectin (PNA) 689:717 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 4 58 theme hypogaea 697:704 arg1 PNA 714:716 PNA 714:716 PNA 714:716 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 4 59 theme POMGNT1 625:631 arg1 digestion 612:620 sialidase digestion 602:620 sialidase digestion of POMGNT1 602:631 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 6 60 theme mass 930:933 arg1 spectrometry 935:946 mass spectrometry 930:946 mass spectrometry 930:946 Next, we analyzed the structures of the O-glycans on POMGNT1 by β-elimination and pyrazolone-labeling methods in combination with mass spectrometry. 26328495 1 61 theme Protein 49:55 arg1 β1,2-N-acetylglucosaminyltransferase 74:109 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 49:111 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) 49:121 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 61 theme Protein 49:55 arg1 POMGNT1 114:120 POMGNT1 114:120 POMGNT1 114:120 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 1 61 theme Protein 49:55 arg1 glycosyltransferase 134:152 a Golgi glycosyltransferase 126:152 a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan 126:254 Protein O-linked mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGNT1) is a Golgi glycosyltransferase that catalyzes the formation of the N-acetylglucosamine (GlcNAc) β1→2Man linkage of O-mannosyl glycan. 26328495 2 62 mod modified 272:279 arg3 N-glycans 284:292 N-glycans 284:292 N-glycans 284:292 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 2 62 mod modified 272:279 arg1 POMGNT1 257:263 POMGNT1 257:263 POMGNT1 257:263 POMGNT1 is not modified by N-glycans because there are no potential N-glycosylation sites; however, it is not clear whether POMGNT1 is modified by O-glycans. 26328495 4 63 theme Arachis 689:695 arg1 lectin 706:711 Arachis hypogaea lectin 689:711 Arachis hypogaea lectin (PNA) 689:717 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 4 63 theme Arachis 689:695 arg1 PNA 714:716 PNA 714:716 PNA 714:716 The recombinant POMGNT1 was recognized by Sambucus sieboldiana lectin (SSA), and sialidase digestion of POMGNT1 decreased SSA reactivity and enhanced the reactivity of Arachis hypogaea lectin (PNA). 26328495 8 64 theme surface 1262:1268 arg1 hydrophobicity 1270:1283 surface hydrophobicity 1262:1283 surface hydrophobicity 1262:1283 To examine whether the O-glycans affect the functions and properties of POMGNT1, we compared glycosylated and non-glycosylated forms of recombinant sPOMGNT1 for their activity and surface hydrophobicity using the hydrophobic probe 1-anilino-8-naphthalene sulfonate (ANS). 24473128 12 0 theme health 1919:1924 arg1 workers 1931:1937 health care workers 1919:1937 health care workers 1919:1937 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 7 1 theme endosomal 978:986 arg1 NPC1 998:1001 NPC1 998:1001 NPC1 998:1001 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 7 1 theme endosomal 978:986 arg1 receptor 988:995 the endosomal receptor 974:995 the endosomal receptor 974:995 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 6 2 theme pseudovirion 755:766 arg1 transduction 768:779 pseudovirion transduction 755:779 pseudovirion transduction 755:779 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 9 3 theme EBOV 1285:1288 arg1 entry 1290:1294 EBOV entry 1285:1294 EBOV entry into murine peritoneal macrophages 1285:1329 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 3 4 contain contains 334:341 arg2 glycans 357:363 only N-linked glycans 343:363 only N-linked glycans 343:363 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 3 4 contain contains 334:341 arg1 cap 330:332 The glycan cap 319:332 The glycan cap 319:332 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 2 5 gly glycosylated 249:260 arg1 domains 262:268 two heavily glycosylated domains 237:268 two heavily glycosylated domains 237:268 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 5 6 theme N-glycosylation 597:611 arg1 sites 613:617 All 15 N-glycosylation sites 590:617 All 15 N-glycosylation sites of EBOV GP1 590:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 5 6 theme N-glycosylation 597:611 arg1 GP1 627:629 EBOV GP1 622:629 EBOV GP1 622:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 15 7 theme envelope 2173:2180 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 7 theme envelope 2173:2180 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 16 8 theme this 2350:2353 arg1 disruption 2336:2345 disruption 2336:2345 disruption of this shield through targeted mutagenesis 2336:2389 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 14 9 theme filovirus 2069:2077 arg1 vaccines 2079:2086 vaccines 2079:2086 vaccines 2079:2086 While filovirus vaccines and therapeutics are being developed, there are no licensed products. 24473128 7 10 theme cap/MLD 1031:1037 arg1 domains 1039:1045 the glycan cap/MLD domains 1020:1045 the glycan cap/MLD domains 1020:1045 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 6 11 theme glycosylation 712:724 arg1 sites 726:730 these 15 glycosylation sites 703:730 these 15 glycosylation sites 703:730 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 9 12 theme peritoneal 1308:1317 arg1 macrophages 1319:1329 murine peritoneal macrophages 1301:1329 murine peritoneal macrophages 1301:1329 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 17 13 theme glycoprotein 2632:2643 arg1 glycosylation 2645:2657 glycoprotein glycosylation 2632:2657 glycoprotein glycosylation 2632:2657 Therefore, our studies provide evidence that filoviruses maintain glycoprotein glycosylation to protect against proteases and antibody neutralization at the expense of efficient entry. 24473128 5 14 gly N-glycosylation 597:611 arg2 sites 613:617 All 15 N-glycosylation sites 590:617 All 15 N-glycosylation sites of EBOV GP1 590:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 5 14 gly N-glycosylation 597:611 arg1 GP1 627:629 EBOV GP1 622:629 EBOV GP1 622:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 5 14 gly N-glycosylation 597:611 arg2 GP1 627:629 EBOV GP1 622:629 EBOV GP1 622:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 5 14 gly N-glycosylation 597:611 arg2 15 594:595 15 594:595 15 594:595 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 4 15 from function 580:587 arg1 understanding 532:544 an understanding 529:544 an understanding of their role in GP structure and function 529:587 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 8 16 theme N-linked 1118:1125 arg1 glycans 1127:1133 GP1 N-linked glycans 1114:1133 GP1 N-linked glycans 1114:1133 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 6 17 gly glycosylation 712:724 arg2 sites 726:730 these 15 glycosylation sites 703:730 these 15 glycosylation sites 703:730 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 6 17 gly glycosylation 712:724 arg2 15 709:710 15 709:710 15 709:710 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 0 18 link N-linked 37:44 arg1 glycans 46:52 N-linked glycans 37:52 N-linked glycans 37:52 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 8 19 dep -dependent 1144:1153 arg1 C-type 1156:1161 C-type 1156:1161 C-type 1156:1161 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 11 20 theme EBOV 1658:1661 arg1 core 1667:1670 the EBOV GP1 core 1654:1670 the EBOV GP1 core 1654:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 18 21 theme potential 2840:2848 arg1 tactics 2865:2871 potential immune evasion tactics 2840:2871 potential immune evasion tactics of the virus 2840:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 4 22 from structure 566:574 arg1 understanding 532:544 an understanding 529:544 an understanding of their role in GP structure and function 529:587 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 3 23 contain contains 382:389 arg1 MLD 378:380 the MLD 374:380 the MLD 374:380 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 3 23 contain contains 382:389 arg2 glycans 412:418 O-linked glycans 403:418 O-linked glycans 403:418 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 3 23 contain contains 382:389 arg2 N- 396:397 N- 396:397 N- 396:397 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 11 24 theme entry 1766:1770 arg1 efficiency 1772:1781 viral entry efficiency 1760:1781 viral entry efficiency 1760:1781 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 11 25 theme antibody 1688:1695 arg1 neutralization 1697:1710 antibody neutralization 1688:1710 antibody neutralization 1688:1710 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 13 26 theme Ebola 2022:2026 arg1 viruses 2028:2034 Ebola viruses 2022:2034 Ebola viruses 2022:2034 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 9 27 theme N-glycans 1354:1362 arg1 independent 1335:1345 independent 1335:1345 independent 1335:1345 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 3 28 link O-linked 403:410 arg1 glycans 412:418 O-linked glycans 403:418 O-linked glycans 403:418 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 13 29 theme Marburg 2048:2054 arg1 virus 2056:2060 Marburg virus 2048:2060 Marburg virus 2048:2060 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 5 30 theme GP1 627:629 arg1 sites 613:617 All 15 N-glycosylation sites 590:617 All 15 N-glycosylation sites of EBOV GP1 590:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 5 30 theme GP1 627:629 arg1 GP1 627:629 EBOV GP1 622:629 EBOV GP1 622:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 15 31 theme immunogenic 2217:2227 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 31 theme immunogenic 2217:2227 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 2 32 theme GP1 216:218 arg1 subunit 220:226 The GP1 subunit 212:226 The GP1 subunit 212:226 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 9 33 theme CLEC-GP1 1381:1388 arg1 interactions 1399:1410 CLEC-GP1 N-glycan interactions 1381:1410 CLEC-GP1 N-glycan interactions 1381:1410 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 4 34 from role 555:558 arg1 function 580:587 function 580:587 function 580:587 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 4 34 from role 555:558 arg1 structure 566:574 GP structure 563:574 GP structure 563:574 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 13 35 theme unpredictable 1946:1958 arg1 outbreaks 1978:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 16 36 theme intracellular 2537:2549 arg1 NPC1 2560:2563 the intracellular receptor NPC1 2533:2563 the intracellular receptor NPC1 2533:2563 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 0 37 from analysis 25:32 arg1 GP1 69:71 Ebola virus GP1 57:71 Ebola virus GP1 57:71 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 13 38 theme fever 1972:1976 arg1 outbreaks 1978:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 18 39 theme evasion 2857:2863 arg1 tactics 2865:2871 potential immune evasion tactics 2840:2871 potential immune evasion tactics of the virus 2840:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 8 40 link N-linked 1118:1125 arg1 glycans 1127:1133 GP1 N-linked glycans 1114:1133 GP1 N-linked glycans 1114:1133 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 17 41 theme entry 2744:2748 arg1 expense 2723:2729 the expense 2719:2729 the expense of efficient entry 2719:2748 Therefore, our studies provide evidence that filoviruses maintain glycoprotein glycosylation to protect against proteases and antibody neutralization at the expense of efficient entry. 24473128 15 42 gly glycoprotein 2182:2193 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 42 gly glycoprotein 2182:2193 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 11 43 theme N-linked 1634:1641 arg1 glycans 1643:1649 the conserved N-linked glycans 1620:1649 the conserved N-linked glycans on the EBOV GP1 core 1620:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 12 44 theme Filovirus 1796:1804 arg1 outbreaks 1806:1814 Filovirus outbreaks 1796:1814 Filovirus outbreaks 1796:1814 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 7 45 with interaction 957:967 arg1 NPC1 998:1001 NPC1 998:1001 NPC1 998:1001 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 7 45 with interaction 957:967 arg1 receptor 988:995 the endosomal receptor 974:995 the endosomal receptor 974:995 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 3 46 dep contains 334:341 arg1 whereas 366:372 whereas 366:372 whereas 366:372 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 16 47 from increase 2403:2410 arg1 sensitivity 2436:2446 protease sensitivity 2427:2446 protease sensitivity 2427:2446 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 16 47 from increase 2403:2410 arg1 sensitivity 2472:2482 antiserum/antibody sensitivity 2453:2482 antiserum/antibody sensitivity 2453:2482 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 16 47 from increase 2403:2410 arg1 entry 2420:2424 cell entry 2415:2424 cell entry 2415:2424 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 15 48 theme glycans 2264:2270 arg1 shield 2254:2259 shield 2254:2259 shield 2254:2259 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 7 49 theme receptor-binding 883:898 arg1 RBD 908:910 RBD 908:910 RBD 908:910 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 7 49 theme receptor-binding 883:898 arg1 domain 900:905 the receptor-binding domain 879:905 the receptor-binding domain (RBD) 879:911 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 1 50 theme Ebola 74:78 arg1 EBOV 87:90 EBOV 87:90 EBOV 87:90 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 1 50 theme Ebola 74:78 arg1 virus 80:84 Ebola virus 74:84 Ebola virus (EBOV) entry 74:97 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 3 51 theme glycan 323:328 arg1 cap 330:332 The glycan cap 319:332 The glycan cap 319:332 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 1 52 theme virion 112:117 arg1 glycoprotein 138:149 the virion surface-associated glycoprotein 108:149 the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2) 108:209 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 1 52 theme virion 112:117 arg1 GP 152:153 GP 152:153 GP 152:153 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 11 53 theme viral 1760:1764 arg1 efficiency 1772:1781 viral entry efficiency 1760:1781 viral entry efficiency 1760:1781 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 0 54 theme Comprehensive 0:12 arg1 analysis 25:32 Comprehensive functional analysis 0:32 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 0:72 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 2 55 contain contains 228:235 arg2 cap 282:284 the glycan cap 271:284 the glycan cap 271:284 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 2 55 contain contains 228:235 arg2 domains 262:268 two heavily glycosylated domains 237:268 two heavily glycosylated domains 237:268 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 2 55 contain contains 228:235 arg2 MLD 313:315 MLD 313:315 MLD 313:315 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 2 55 contain contains 228:235 arg2 domain 305:310 the mucin-like domain 290:310 the mucin-like domain (MLD) 290:316 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 2 55 contain contains 228:235 arg1 subunit 220:226 The GP1 subunit 212:226 The GP1 subunit 212:226 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 18 56 theme immune 2850:2855 arg1 tactics 2865:2871 potential immune evasion tactics 2840:2871 potential immune evasion tactics of the virus 2840:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 9 57 theme important 1449:1457 arg1 cell 1467:1470 this important primary cell 1444:1470 this important primary cell 1444:1470 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 10 58 theme antibody 1554:1561 arg1 sensitivity 1563:1573 antibody sensitivity 1554:1573 antibody sensitivity 1554:1573 Finally, the removal of all GP1 N-glycans outside the MLD enhanced antiserum and antibody sensitivity. 24473128 1 59 theme surface-associated 119:136 arg1 glycoprotein 138:149 the virion surface-associated glycoprotein 108:149 the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2) 108:209 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 1 59 theme surface-associated 119:136 arg1 GP 152:153 GP 152:153 GP 152:153 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 3 60 theme N-linked 348:355 arg1 glycans 357:363 only N-linked glycans 343:363 only N-linked glycans 343:363 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 4 61 theme glycan 508:513 arg1 sites 515:519 N-linked glycan sites 499:519 N-linked glycan sites 499:519 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 0 62 theme N-linked 37:44 arg1 glycans 46:52 N-linked glycans 37:52 N-linked glycans 37:52 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 4 63 theme EBOV 464:467 arg1 GP1 469:471 EBOV GP1 464:471 EBOV GP1 464:471 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 0 64 theme virus 63:67 arg1 GP1 69:71 Ebola virus GP1 57:71 Ebola virus GP1 57:71 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 2 65 theme mucin-like 294:303 arg1 MLD 313:315 MLD 313:315 MLD 313:315 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 2 65 theme mucin-like 294:303 arg1 domain 305:310 the mucin-like domain 290:310 the mucin-like domain (MLD) 290:316 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 3 66 theme O-linked 403:410 arg1 glycans 412:418 O-linked glycans 403:418 O-linked glycans 403:418 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 1 67 theme heterodimers 188:199 arg1 trimer 178:183 a trimer 176:183 a trimer of heterodimers (GP1/GP2) 176:209 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 8 68 from effects 1091:1097 arg1 transduction 1188:1199 Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction 1138:1199 Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction 1138:1199 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 8 69 dep -dependent 1177:1186 arg1 -dependent 1144:1153 -dependent 1144:1153 -dependent 1144:1153 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 16 70 theme receptor 2551:2558 arg1 NPC1 2560:2563 the intracellular receptor NPC1 2533:2563 the intracellular receptor NPC1 2533:2563 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 18 71 theme virus 2880:2884 arg1 tactics 2865:2871 potential immune evasion tactics 2840:2871 potential immune evasion tactics of the virus 2840:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 18 71 theme virus 2880:2884 arg1 filoviruses 2824:2834 filoviruses 2824:2834 filoviruses 2824:2834 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 12 72 theme general 1900:1906 arg1 public 1908:1913 the general public 1896:1913 the general public 1896:1913 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 4 73 theme role 555:558 arg1 understanding 532:544 an understanding 529:544 an understanding of their role in GP structure and function 529:587 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 7 74 theme RBD 1052:1054 arg1 residues 1056:1063 RBD residues 1052:1063 RBD residues required for binding 1052:1084 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 4 75 link N-linked 499:506 arg1 sites 515:519 N-linked glycan sites 499:519 N-linked glycan sites 499:519 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 6 76 from increase 821:828 arg1 sensitivity 842:852 protease sensitivity 833:852 protease sensitivity 833:852 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 2 77 theme glycosylated 249:260 arg1 domains 262:268 two heavily glycosylated domains 237:268 two heavily glycosylated domains 237:268 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 15 78 theme viral 2167:2171 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 78 theme viral 2167:2171 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 12 79 theme fatality 1875:1882 arg1 rates 1884:1888 high fatality rates 1870:1888 high fatality rates 1870:1888 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 6 80 theme protease 833:840 arg1 sensitivity 842:852 protease sensitivity 833:852 protease sensitivity 833:852 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 11 81 theme negative 1724:1731 arg1 impact 1733:1738 the negative impact 1720:1738 the negative impact the glycans have on viral entry efficiency 1720:1781 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 7 82 theme glycan 1024:1029 arg1 domains 1039:1045 the glycan cap/MLD domains 1020:1045 the glycan cap/MLD domains 1020:1045 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 6 83 theme sites 726:730 arg1 loss 695:698 The loss 691:698 The loss of these 15 glycosylation sites 691:730 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 9 84 theme murine 1301:1306 arg1 macrophages 1319:1329 murine peritoneal macrophages 1301:1329 murine peritoneal macrophages 1301:1329 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 16 85 theme targeted 2370:2377 arg1 mutagenesis 2379:2389 targeted mutagenesis 2370:2389 targeted mutagenesis 2370:2389 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 2 86 theme glycan 275:280 arg1 cap 282:284 the glycan cap 271:284 the glycan cap 271:284 The GP1 subunit contains two heavily glycosylated domains, the glycan cap and the mucin-like domain (MLD). 24473128 8 87 theme loss 1106:1109 arg1 effects 1091:1097 The effects 1087:1097 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction 1087:1199 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 8 87 theme loss 1106:1109 arg1 complex 1206:1212 complex 1206:1212 complex 1206:1212 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 8 88 theme GP1 1114:1116 arg1 glycans 1127:1133 GP1 N-linked glycans 1114:1133 GP1 N-linked glycans 1114:1133 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 11 89 from glycans 1643:1649 arg1 core 1667:1670 the EBOV GP1 core 1654:1670 the EBOV GP1 core 1654:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 4 90 theme GP 563:564 arg1 structure 566:574 GP structure 563:574 GP structure 563:574 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 10 91 theme GP1 1501:1503 arg1 N-glycans 1505:1513 all GP1 N-glycans 1497:1513 all GP1 N-glycans 1497:1513 Finally, the removal of all GP1 N-glycans outside the MLD enhanced antiserum and antibody sensitivity. 24473128 11 92 theme GP1 1663:1665 arg1 core 1667:1670 the EBOV GP1 core 1654:1670 the EBOV GP1 core 1654:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 9 93 theme GP1 1350:1352 arg1 N-glycans 1354:1362 GP1 N-glycans 1350:1362 GP1 N-glycans 1350:1362 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 8 94 theme glycans 1127:1133 arg1 loss 1106:1109 the loss 1102:1109 the loss of GP1 N-linked glycans 1102:1133 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 16 95 theme virion 2515:2520 arg1 binding 2522:2528 virion binding 2515:2528 virion binding to the intracellular receptor NPC1 2515:2563 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 12 96 theme high 1870:1873 arg1 rates 1884:1888 high fatality rates 1870:1888 high fatality rates 1870:1888 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 17 97 gly glycoprotein 2632:2643 arg1 glycoprotein 2632:2643 glycoprotein glycosylation 2632:2657 glycoprotein glycosylation 2632:2657 Therefore, our studies provide evidence that filoviruses maintain glycoprotein glycosylation to protect against proteases and antibody neutralization at the expense of efficient entry. 24473128 1 98 gly glycoprotein 138:149 arg1 glycoprotein 138:149 the virion surface-associated glycoprotein 108:149 the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2) 108:209 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 1 98 gly glycoprotein 138:149 arg1 GP 152:153 GP 152:153 GP 152:153 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 13 99 theme viruses 2028:2034 arg1 species 2011:2017 multiple species 2002:2017 multiple species of Ebola viruses, as well as Marburg virus 2002:2060 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 3 100 link N-linked 348:355 arg1 glycans 357:363 only N-linked glycans 343:363 only N-linked glycans 343:363 The glycan cap contains only N-linked glycans, whereas the MLD contains both N- and O-linked glycans. 24473128 5 101 theme EBOV 622:625 arg1 GP1 627:629 EBOV GP1 622:629 EBOV GP1 622:629 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 18 102 theme tactics 2865:2871 arg1 process 2813:2819 the unique entry process 2796:2819 the unique entry process of filoviruses and potential immune evasion tactics of the virus 2796:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 16 103 theme protease 2427:2434 arg1 sensitivity 2436:2446 protease sensitivity 2427:2446 protease sensitivity 2427:2446 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 8 104 theme -dependent 1177:1186 arg1 transduction 1188:1199 Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction 1138:1199 Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction 1138:1199 The effects of the loss of GP1 N-linked glycans on Ca(2+)-dependent (C-type) lectin (CLEC)-dependent transduction were complex, and the effect was unique for each of the CLECs tested. 24473128 11 105 theme conserved 1624:1632 arg1 glycans 1643:1649 the conserved N-linked glycans 1620:1649 the conserved N-linked glycans on the EBOV GP1 core 1620:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 5 106 theme GP 687:688 arg1 expression 673:682 the expression 669:682 the expression of GP 669:688 All 15 N-glycosylation sites of EBOV GP1 could be removed without compromising the expression of GP. 24473128 13 107 theme multiple 2002:2009 arg1 species 2011:2017 multiple species 2002:2017 multiple species of Ebola viruses, as well as Marburg virus 2002:2060 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 9 108 theme N-glycan 1390:1397 arg1 interactions 1399:1410 CLEC-GP1 N-glycan interactions 1381:1410 CLEC-GP1 N-glycan interactions 1381:1410 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 12 109 theme care 1926:1929 arg1 workers 1931:1937 health care workers 1919:1937 health care workers 1919:1937 IMPORTANCE: Filovirus outbreaks occur sporadically throughout central Africa, causing high fatality rates among the general public and health care workers. 24473128 13 110 theme hemorrhagic 1960:1970 arg1 outbreaks 1978:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks 1940:1986 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 16 111 theme antiserum/antibody 2453:2470 arg1 sensitivity 2472:2482 antiserum/antibody sensitivity 2453:2482 antiserum/antibody sensitivity 2453:2482 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 17 112 theme efficient 2734:2742 arg1 entry 2744:2748 efficient entry 2734:2748 efficient entry 2734:2748 Therefore, our studies provide evidence that filoviruses maintain glycoprotein glycosylation to protect against proteases and antibody neutralization at the expense of efficient entry. 24473128 11 113 link N-linked 1634:1641 arg1 glycans 1643:1649 the conserved N-linked glycans 1620:1649 the conserved N-linked glycans on the EBOV GP1 core 1620:1670 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 4 114 from understanding 532:544 arg1 function 580:587 function 580:587 function 580:587 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 4 114 from understanding 532:544 arg1 structure 566:574 GP structure 563:574 GP structure 563:574 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 16 115 dep this 2350:2353 arg1 shield 2355:2360 shield 2355:2360 shield through targeted mutagenesis 2355:2389 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 24473128 18 116 theme interesting 2770:2780 arg1 insights 2782:2789 interesting insights 2770:2789 interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus 2770:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 15 117 theme conserved 2288:2296 arg1 domain 2315:2320 the conserved receptor-binding domain 2284:2320 the conserved receptor-binding domain 2284:2320 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 4 118 theme Site-directed 421:433 arg1 mutagenesis 435:445 Site-directed mutagenesis 421:445 Site-directed mutagenesis 421:445 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 15 119 theme principal 2207:2215 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 119 theme principal 2207:2215 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 0 120 theme functional 14:23 arg1 analysis 25:32 Comprehensive functional analysis 0:32 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 0:72 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 1 121 theme virus 80:84 arg1 entry 93:97 Ebola virus (EBOV) entry 74:97 Ebola virus (EBOV) entry 74:97 Ebola virus (EBOV) entry requires the virion surface-associated glycoprotein (GP) that is composed of a trimer of heterodimers (GP1/GP2). 24473128 18 122 theme entry 2807:2811 arg1 process 2813:2819 the unique entry process 2796:2819 the unique entry process of filoviruses and potential immune evasion tactics of the virus 2796:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 15 123 contain contains 2237:2244 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 123 contain contains 2237:2244 arg2 shield 2254:2259 shield 2254:2259 shield 2254:2259 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 123 contain contains 2237:2244 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 4 124 theme N-linked 499:506 arg1 sites 515:519 N-linked glycan sites 499:519 N-linked glycan sites 499:519 Site-directed mutagenesis was performed on EBOV GP1 to systematically disrupt N-linked glycan sites to gain an understanding of their role in GP structure and function. 24473128 14 125 theme licensed 2139:2146 arg1 products 2148:2155 no licensed products 2136:2155 no licensed products 2136:2155 While filovirus vaccines and therapeutics are being developed, there are no licensed products. 24473128 10 126 theme N-glycans 1505:1513 arg1 removal 1486:1492 the removal 1482:1492 the removal of all GP1 N-glycans outside the MLD 1482:1529 Finally, the removal of all GP1 N-glycans outside the MLD enhanced antiserum and antibody sensitivity. 24473128 15 127 theme sole 2162:2165 arg1 glycoprotein 2182:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein 2158:2193 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 15 127 theme sole 2162:2165 arg1 target 2229:2234 a principal immunogenic target 2205:2234 a principal immunogenic target 2205:2234 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 9 128 theme primary 1459:1465 arg1 cell 1467:1470 this important primary cell 1444:1470 this important primary cell 1444:1470 Surprisingly, EBOV entry into murine peritoneal macrophages was independent of GP1 N-glycans, suggesting that CLEC-GP1 N-glycan interactions are not required for entry into this important primary cell. 24473128 7 129 dep glycan 929:934 arg1 shield 936:941 shield 936:941 shield 936:941 Interestingly, exposing the receptor-binding domain (RBD) by removing the glycan shield did not allow interaction with the endosomal receptor, NPC1, indicating that the glycan cap/MLD domains mask RBD residues required for binding. 24473128 0 130 theme glycans 46:52 arg1 analysis 25:32 Comprehensive functional analysis 0:32 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 0:72 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 17 131 theme antibody 2692:2699 arg1 neutralization 2701:2714 antibody neutralization 2692:2714 antibody neutralization 2692:2714 Therefore, our studies provide evidence that filoviruses maintain glycoprotein glycosylation to protect against proteases and antibody neutralization at the expense of efficient entry. 24473128 0 132 theme Ebola 57:61 arg1 GP1 69:71 Ebola virus GP1 57:71 Ebola virus GP1 57:71 Comprehensive functional analysis of N-linked glycans on Ebola virus GP1. 24473128 18 133 theme unique 2800:2805 arg1 process 2813:2819 the unique entry process 2796:2819 the unique entry process of filoviruses and potential immune evasion tactics of the virus 2796:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 15 134 theme receptor-binding 2298:2313 arg1 domain 2315:2320 the conserved receptor-binding domain 2284:2320 the conserved receptor-binding domain 2284:2320 The sole viral envelope glycoprotein, which is a principal immunogenic target, contains a heavy shield of glycans surrounding the conserved receptor-binding domain. 24473128 18 135 theme filoviruses 2824:2834 arg1 process 2813:2819 the unique entry process 2796:2819 the unique entry process of filoviruses and potential immune evasion tactics of the virus 2796:2884 Our results unveil interesting insights into the unique entry process of filoviruses and potential immune evasion tactics of the virus. 24473128 6 136 theme Vero 784:787 arg1 cells 789:793 Vero cells 784:793 Vero cells 784:793 The loss of these 15 glycosylation sites significantly enhanced pseudovirion transduction in Vero cells, which correlated with an increase in protease sensitivity. 24473128 11 137 dep impact 1733:1738 arg1 have 1752:1755 have 1752:1755 have on viral entry efficiency 1752:1781 In total, our results provide evidence that the conserved N-linked glycans on the EBOV GP1 core protect GP from antibody neutralization despite the negative impact the glycans have on viral entry efficiency. 24473128 13 138 theme virus 2056:2060 arg1 species 2011:2017 multiple species 2002:2017 multiple species of Ebola viruses, as well as Marburg virus 2002:2060 These unpredictable hemorrhagic fever outbreaks are caused by multiple species of Ebola viruses, as well as Marburg virus. 24473128 16 139 theme cell 2415:2418 arg1 entry 2420:2424 cell entry 2415:2424 cell entry 2415:2424 We find that disruption of this shield through targeted mutagenesis leads to an increase in cell entry, protease sensitivity, and antiserum/antibody sensitivity but is not sufficient to allow virion binding to the intracellular receptor NPC1. 28890362 2 0 theme protein 469:475 arg1 surface 477:483 the Env protein surface 461:483 the Env protein surface 461:483 Here we use molecular dynamics to provide insight into its structural dynamics and into how both protomer and glycan movements coordinate to shield the Env protein surface. 28890362 6 1 theme glycan 1048:1053 arg1 patches 1037:1043 patches 1037:1043 patches of glycan 1037:1053 We also observed stable microdomains comprising patches of glycan, with neutralizing antibodies generally binding at the interface between glycan microdomains. 28890362 0 2 theme Trimer 65:70 arg1 Dynamics 12:19 Microsecond Dynamics 0:19 Microsecond Dynamics 0:19 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 0 2 theme Trimer 65:70 arg1 Analysis 33:40 Network Analysis 25:40 Network Analysis 25:40 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 1 3 gly glycosylated 207:218 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 4 theme trimeric 152:159 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 4 theme trimeric 152:159 arg1 one 191:193 one 191:193 one 191:193 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 4 theme trimeric 152:159 arg1 spike 182:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 5 5 theme high-mannose 951:962 arg1 character 964:972 the high-mannose character 947:972 the high-mannose character of the shield 947:986 The network property of betweenness centrality appeared to identify whether glycans spread to restrict access or cluster to maintain the high-mannose character of the shield. 28890362 7 6 dep glycan 1223:1228 arg1 shield 1230:1235 shield 1230:1235 shield 1230:1235 Overall, our results provide a microsecond-based understanding of the Env glycan shield. 28890362 4 7 theme binding 780:786 arg1 receptor 804:811 the CD4 receptor 796:811 the CD4 receptor 796:811 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 4 7 theme binding 780:786 arg1 site 788:791 the binding site 776:791 the binding site of the CD4 receptor 776:811 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 0 8 theme Collective 79:88 arg1 Behavior 90:97 Collective Behavior 79:97 Collective Behavior 79:97 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 6 9 theme stable 1006:1011 arg1 microdomains 1013:1024 stable microdomains 1006:1024 stable microdomains comprising patches of glycan 1006:1053 We also observed stable microdomains comprising patches of glycan, with neutralizing antibodies generally binding at the interface between glycan microdomains. 28890362 0 10 theme Conserved 103:111 arg1 Microdomains 113:124 Conserved Microdomains 103:124 Conserved Microdomains 103:124 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 0 10 theme Conserved 103:111 arg1 Shield 140:145 Shield 140:145 Shield 140:145 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 1 11 theme HIV-1-envelope 161:174 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 11 theme HIV-1-envelope 161:174 arg1 one 191:193 one 191:193 one 191:193 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 11 theme HIV-1-envelope 161:174 arg1 spike 182:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 2 12 theme glycan 423:428 arg1 movements 430:438 glycan movements 423:438 glycan movements 423:438 Here we use molecular dynamics to provide insight into its structural dynamics and into how both protomer and glycan movements coordinate to shield the Env protein surface. 28890362 6 13 theme glycan 1128:1133 arg1 microdomains 1135:1146 glycan microdomains 1128:1146 glycan microdomains 1128:1146 We also observed stable microdomains comprising patches of glycan, with neutralizing antibodies generally binding at the interface between glycan microdomains. 28890362 2 14 theme structural 372:381 arg1 dynamics 383:390 its structural dynamics 368:390 its structural dynamics 368:390 Here we use molecular dynamics to provide insight into its structural dynamics and into how both protomer and glycan movements coordinate to shield the Env protein surface. 28890362 7 15 theme glycan 1223:1228 arg1 understanding 1198:1210 a microsecond-based understanding 1178:1210 a microsecond-based understanding of the Env glycan shield 1178:1235 Overall, our results provide a microsecond-based understanding of the Env glycan shield. 28890362 2 16 theme Env 465:467 arg1 surface 477:483 the Env protein surface 461:483 the Env protein surface 461:483 Here we use molecular dynamics to provide insight into its structural dynamics and into how both protomer and glycan movements coordinate to shield the Env protein surface. 28890362 3 17 theme model 557:561 arg1 simulation 512:521 A 2-μs molecular dynamics simulation 486:521 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env 486:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 1 18 theme Env 177:179 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 18 theme Env 177:179 arg1 one 191:193 one 191:193 one 191:193 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 18 theme Env 177:179 arg1 spike 182:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 0 19 theme Microsecond 0:10 arg1 Dynamics 12:19 Microsecond Dynamics 0:19 Microsecond Dynamics 0:19 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 1 20 theme half 258:261 arg1 mass 267:270 roughly half its mass 250:270 roughly half its mass comprising host-derived N-linked glycan 250:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 4 21 theme conserved 702:710 arg1 glycans 712:718 highly conserved glycans 695:718 highly conserved glycans combined with protomer 695:741 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 0 22 theme Network 25:31 arg1 Analysis 33:40 Network Analysis 25:40 Network Analysis 25:40 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 5 23 theme network 818:824 arg1 property 826:833 The network property 814:833 The network property of betweenness centrality 814:859 The network property of betweenness centrality appeared to identify whether glycans spread to restrict access or cluster to maintain the high-mannose character of the shield. 28890362 3 24 theme glycosylated 534:545 arg1 model 557:561 a fully glycosylated atomistic model 526:561 a fully glycosylated atomistic model of the HIV-1 SOSIP Env 526:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 5 25 theme betweenness 838:848 arg1 centrality 850:859 betweenness centrality 838:859 betweenness centrality 838:859 The network property of betweenness centrality appeared to identify whether glycans spread to restrict access or cluster to maintain the high-mannose character of the shield. 28890362 7 26 theme Env 1219:1221 arg1 glycan 1223:1228 the Env glycan shield 1215:1235 the Env glycan shield 1215:1235 Overall, our results provide a microsecond-based understanding of the Env glycan shield. 28890362 3 27 theme trimer-opening 640:653 arg1 movements 655:663 protomer-scissoring and trimer-opening movements 616:663 protomer-scissoring and trimer-opening movements 616:663 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 3 28 theme atomistic 547:555 arg1 model 557:561 a fully glycosylated atomistic model 526:561 a fully glycosylated atomistic model of the HIV-1 SOSIP Env 526:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 5 29 theme centrality 850:859 arg1 property 826:833 The network property 814:833 The network property of betweenness centrality 814:859 The network property of betweenness centrality appeared to identify whether glycans spread to restrict access or cluster to maintain the high-mannose character of the shield. 28890362 3 30 theme movements 655:663 arg1 spectrum 604:611 a spectrum 602:611 a spectrum of protomer-scissoring and trimer-opening movements 602:663 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 3 31 theme SOSIP 576:580 arg1 Env 582:584 the HIV-1 SOSIP Env 566:584 the HIV-1 SOSIP Env 566:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 4 32 theme Network 666:672 arg1 analysis 674:681 Network analysis 666:681 Network analysis 666:681 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 1 33 theme host-derived 283:294 arg1 glycan 305:310 host-derived N-linked glycan 283:310 host-derived N-linked glycan 283:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 3 34 theme protomer-scissoring 616:634 arg1 movements 655:663 protomer-scissoring and trimer-opening movements 616:663 protomer-scissoring and trimer-opening movements 616:663 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 3 35 theme Env 582:584 arg1 model 557:561 a fully glycosylated atomistic model 526:561 a fully glycosylated atomistic model of the HIV-1 SOSIP Env 526:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 0 36 theme HIV-1 49:53 arg1 Trimer 65:70 the HIV-1 SOSIP Env Trimer 45:70 the HIV-1 SOSIP Env Trimer 45:70 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 1 37 theme N-linked 296:303 arg1 glycan 305:310 host-derived N-linked glycan 283:310 host-derived N-linked glycan 283:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 2 38 theme molecular 325:333 arg1 dynamics 335:342 molecular dynamics 325:342 molecular dynamics 325:342 Here we use molecular dynamics to provide insight into its structural dynamics and into how both protomer and glycan movements coordinate to shield the Env protein surface. 28890362 7 39 theme microsecond-based 1180:1196 arg1 understanding 1198:1210 a microsecond-based understanding 1178:1210 a microsecond-based understanding of the Env glycan shield 1178:1235 Overall, our results provide a microsecond-based understanding of the Env glycan shield. 28890362 0 40 theme Shield 140:145 arg1 Behavior 90:97 Collective Behavior 79:97 Collective Behavior 79:97 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 0 40 theme Shield 140:145 arg1 Microdomains 113:124 Conserved Microdomains 103:124 Conserved Microdomains 103:124 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 0 40 theme Shield 140:145 arg1 Shield 140:145 Shield 140:145 Shield 140:145 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 3 41 theme HIV-1 570:574 arg1 Env 582:584 the HIV-1 SOSIP Env 566:584 the HIV-1 SOSIP Env 566:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 1 42 link host-derived 283:294 arg1 glycan 305:310 host-derived N-linked glycan 283:310 host-derived N-linked glycan 283:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 3 43 theme 2-μs 488:491 arg1 simulation 512:521 A 2-μs molecular dynamics simulation 486:521 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env 486:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 4 44 theme CD4 800:802 arg1 receptor 804:811 the CD4 receptor 796:811 the CD4 receptor 796:811 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 1 45 link N-linked 296:303 arg1 glycan 305:310 host-derived N-linked glycan 283:310 host-derived N-linked glycan 283:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 46 theme the most 198:205 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 5 47 theme shield 981:986 arg1 character 964:972 the high-mannose character 947:972 the high-mannose character of the shield 947:986 The network property of betweenness centrality appeared to identify whether glycans spread to restrict access or cluster to maintain the high-mannose character of the shield. 28890362 3 48 gly glycosylated 534:545 arg1 model 557:561 a fully glycosylated atomistic model 526:561 a fully glycosylated atomistic model of the HIV-1 SOSIP Env 526:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 1 49 theme glycosylated 207:218 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 0 50 theme Env 61:63 arg1 Trimer 65:70 the HIV-1 SOSIP Env Trimer 45:70 the HIV-1 SOSIP Env Trimer 45:70 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 1 51 with complexes 228:236 arg1 mass 267:270 roughly half its mass 250:270 roughly half its mass comprising host-derived N-linked glycan 250:310 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 3 52 theme molecular 493:501 arg1 simulation 512:521 A 2-μs molecular dynamics simulation 486:521 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env 486:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 1 53 theme protein 220:226 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 0 54 theme SOSIP 55:59 arg1 Trimer 65:70 the HIV-1 SOSIP Env Trimer 45:70 the HIV-1 SOSIP Env Trimer 45:70 Microsecond Dynamics and Network Analysis of the HIV-1 SOSIP Env Trimer Reveal Collective Behavior and Conserved Microdomains of the Glycan Shield. 28890362 4 55 theme receptor 804:811 arg1 receptor 804:811 the CD4 receptor 796:811 the CD4 receptor 796:811 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 4 55 theme receptor 804:811 arg1 site 788:791 the binding site 776:791 the binding site of the CD4 receptor 776:811 Network analysis showed that highly conserved glycans combined with protomer scissoring to restrict access to the binding site of the CD4 receptor. 28890362 3 56 theme dynamics 503:510 arg1 simulation 512:521 A 2-μs molecular dynamics simulation 486:521 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env 486:584 A 2-μs molecular dynamics simulation of a fully glycosylated atomistic model of the HIV-1 SOSIP Env trimer revealed a spectrum of protomer-scissoring and trimer-opening movements. 28890362 1 57 theme complexes 228:236 arg1 complexes 228:236 the most glycosylated protein complexes 198:236 the most glycosylated protein complexes 198:236 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 57 theme complexes 228:236 arg1 one 191:193 one 191:193 one 191:193 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28890362 1 57 theme complexes 228:236 arg1 spike 182:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike 148:186 The trimeric HIV-1-envelope (Env) spike is one of the most glycosylated protein complexes known, with roughly half its mass comprising host-derived N-linked glycan. 28065507 2 0 from sucrose 449:455 arg1 polysaccharides 428:442 α-glucan-type extracellular polysaccharides 400:442 α-glucan-type extracellular polysaccharides 400:442 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 3 1 theme crystal 549:555 arg1 structure 557:565 The crystal structure 545:565 The crystal structure of a 4,6-α-glucanotransferase 545:595 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 2 2 theme GH70 513:516 arg1 4,6-α-glucanotransferases 518:542 GH70 4,6-α-glucanotransferases 513:542 GH70 4,6-α-glucanotransferases 513:542 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 5 3 theme in vivo 991:997 arg1 studies 999:1005 genomic, phylogenetic, and in vivo studies 964:1005 genomic, phylogenetic, and in vivo studies 964:1005 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 4 theme 4,6-α-GTs 1147:1155 arg1 evolution 1134:1142 the evolution 1130:1142 the evolution of 4,6-α-GTs and glucansucrases from α-amylases 1130:1190 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 0 5 theme Oral 112:115 arg1 Bacteria 117:124 Oral Bacteria 112:124 Oral Bacteria 112:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 5 6 theme extracellular 1227:1239 arg1 polymers 1241:1248 extracellular polymers 1227:1248 extracellular polymers that contribute to biofilm formation from different substrates 1227:1311 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 7 theme critical 1106:1113 arg1 factors 1115:1121 critical factors 1106:1121 critical factors 1106:1121 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 1 8 theme enzymes 262:268 arg1 evolution 249:257 the evolution 245:257 the evolution of enzymes in human microbiota 245:288 Food processing and refining has dramatically changed the human diet, but little is known about whether this affected the evolution of enzymes in human microbiota. 28065507 1 9 theme in human 270:277 arg1 microbiota 279:288 in human microbiota 270:288 enzymes in human microbiota 262:288 Food processing and refining has dramatically changed the human diet, but little is known about whether this affected the evolution of enzymes in human microbiota. 28065507 0 10 from Evolution 67:75 arg1 α-Amylases 98:107 α-Amylases 98:107 α-Amylases in Oral Bacteria 98:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 3 11 theme 4,6-α-glucanotransferase 572:595 arg1 structure 557:565 The crystal structure 545:565 The crystal structure of a 4,6-α-glucanotransferase 545:595 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 1 12 theme human 185:189 arg1 diet 191:194 the human diet 181:194 the human diet 181:194 Food processing and refining has dramatically changed the human diet, but little is known about whether this affected the evolution of enzymes in human microbiota. 28065507 5 13 theme starch 1062:1067 arg1 derivatives 1069:1079 starch derivatives 1062:1079 starch derivatives 1062:1079 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 2 14 theme α-glucan-type 400:412 arg1 polysaccharides 428:442 α-glucan-type extracellular polysaccharides 400:442 α-glucan-type extracellular polysaccharides 400:442 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 5 15 theme oral 1202:1205 arg1 bacteria 1207:1214 oral bacteria 1202:1214 oral bacteria 1202:1214 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 16 from evolution 1134:1142 arg1 α-amylases 1181:1190 α-amylases 1181:1190 α-amylases 1181:1190 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 2 17 theme hydrolase 326:334 arg1 family 336:341 glycoside hydrolase family 70 316:344 glycoside hydrolase family 70 (GH70) 316:351 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 2 17 theme hydrolase 326:334 arg1 GH70 347:350 GH70 347:350 GH70 347:350 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 5 18 theme genomic 964:970 arg1 studies 999:1005 genomic, phylogenetic, and in vivo studies 964:1005 genomic, phylogenetic, and in vivo studies 964:1005 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 0 19 theme Crystal 0:6 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of 4,6-α-Glucanotransferase 0:44 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 5 20 theme biofilm 1269:1275 arg1 formation 1277:1285 biofilm formation 1269:1285 biofilm formation from different substrates 1269:1311 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 2 21 theme glycoside 316:324 arg1 family 336:341 glycoside hydrolase family 70 316:344 glycoside hydrolase family 70 (GH70) 316:351 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 2 21 theme glycoside 316:324 arg1 GH70 347:350 GH70 347:350 GH70 347:350 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 4 22 theme active-site 734:744 arg1 loops 746:750 active-site loops 734:750 active-site loops 734:750 While the α-amylase substrate-binding scaffold is retained, active-site loops adapted to favor transglycosylation over hydrolysis; the structure also gives clues as to how 4,6-α-glucanotransferases may have evolved further toward sucrose utilization instead of starch. 28065507 0 23 theme 4,6-α-Glucanotransferase 21:44 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of 4,6-α-Glucanotransferase 0:44 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 4 24 theme substrate-binding 694:710 arg1 scaffold 712:719 the α-amylase substrate-binding scaffold 680:719 the α-amylase substrate-binding scaffold 680:719 While the α-amylase substrate-binding scaffold is retained, active-site loops adapted to favor transglycosylation over hydrolysis; the structure also gives clues as to how 4,6-α-glucanotransferases may have evolved further toward sucrose utilization instead of starch. 28065507 3 25 theme unique 629:634 arg1 specificity 644:654 unique product specificity 629:654 unique product specificity of these enzymes 629:671 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 5 26 theme dietary 1024:1030 arg1 changes 1032:1038 dietary changes 1024:1038 dietary changes involving starch (and starch derivatives) and sucrose intake 1024:1099 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 0 27 from α-Amylases 98:107 arg1 Bacteria 117:124 Oral Bacteria 112:124 Oral Bacteria 112:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 0 27 from α-Amylases 98:107 arg1 Evolution 67:75 Diet-Driven Evolution 55:75 Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria 55:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 0 27 from α-Amylases 98:107 arg1 Enzymes 85:91 GH70 Enzymes 80:91 GH70 Enzymes from α-Amylases in Oral Bacteria 80:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 3 28 theme action 618:623 arg1 mode 610:613 the mode 606:613 the mode of action 606:623 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 3 28 theme action 618:623 arg1 specificity 644:654 unique product specificity 629:654 unique product specificity of these enzymes 629:671 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 4 29 theme α-amylase 684:692 arg1 scaffold 712:719 the α-amylase substrate-binding scaffold 680:719 the α-amylase substrate-binding scaffold 680:719 While the α-amylase substrate-binding scaffold is retained, active-site loops adapted to favor transglycosylation over hydrolysis; the structure also gives clues as to how 4,6-α-glucanotransferases may have evolved further toward sucrose utilization instead of starch. 28065507 0 30 theme Diet-Driven 55:65 arg1 Evolution 67:75 Diet-Driven Evolution 55:75 Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria 55:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 5 31 theme different 1292:1300 arg1 substrates 1302:1311 different substrates 1292:1311 different substrates 1292:1311 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 32 from substrates 1302:1311 arg1 formation 1277:1285 biofilm formation 1269:1285 biofilm formation from different substrates 1269:1311 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 4 33 theme sucrose 904:910 arg1 utilization 912:922 sucrose utilization 904:922 sucrose utilization instead of starch 904:940 While the α-amylase substrate-binding scaffold is retained, active-site loops adapted to favor transglycosylation over hydrolysis; the structure also gives clues as to how 4,6-α-glucanotransferases may have evolved further toward sucrose utilization instead of starch. 28065507 2 34 theme extracellular 414:426 arg1 polysaccharides 428:442 α-glucan-type extracellular polysaccharides 400:442 α-glucan-type extracellular polysaccharides 400:442 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 5 35 theme glucansucrases 1161:1174 arg1 evolution 1134:1142 the evolution 1130:1142 the evolution of 4,6-α-GTs and glucansucrases from α-amylases 1130:1190 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 36 from α-amylases 1181:1190 arg1 evolution 1134:1142 the evolution 1130:1142 the evolution of 4,6-α-GTs and glucansucrases from α-amylases 1130:1190 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 36 from α-amylases 1181:1190 arg1 glucansucrases 1161:1174 glucansucrases 1161:1174 glucansucrases 1161:1174 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 5 36 from α-amylases 1181:1190 arg1 4,6-α-GTs 1147:1155 4,6-α-GTs 1147:1155 4,6-α-GTs 1147:1155 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 3 37 theme product 636:642 arg1 specificity 644:654 unique product specificity 629:654 unique product specificity of these enzymes 629:671 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 1 38 theme Food 127:130 arg1 processing 132:141 Food processing 127:141 Food processing 127:141 Food processing and refining has dramatically changed the human diet, but little is known about whether this affected the evolution of enzymes in human microbiota. 28065507 2 39 theme starch-acting 483:495 arg1 α-amylases 497:506 GH13 starch-acting α-amylases 478:506 GH13 starch-acting α-amylases 478:506 We present evidence that glycoside hydrolase family 70 (GH70) glucansucrases from lactobacilli, synthesizing α-glucan-type extracellular polysaccharides from sucrose, likely evolved from GH13 starch-acting α-amylases, via GH70 4,6-α-glucanotransferases. 28065507 1 40 dep enzymes 262:268 arg1 microbiota 279:288 in human microbiota 270:288 enzymes in human microbiota 262:288 Food processing and refining has dramatically changed the human diet, but little is known about whether this affected the evolution of enzymes in human microbiota. 28065507 0 41 theme Enzymes 85:91 arg1 Evolution 67:75 Diet-Driven Evolution 55:75 Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria 55:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 3 42 theme enzymes 665:671 arg1 mode 610:613 the mode 606:613 the mode of action 606:623 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 3 42 theme enzymes 665:671 arg1 specificity 644:654 unique product specificity 629:654 unique product specificity of these enzymes 629:671 The crystal structure of a 4,6-α-glucanotransferase explains the mode of action and unique product specificity of these enzymes. 28065507 5 43 theme phylogenetic 973:984 arg1 studies 999:1005 genomic, phylogenetic, and in vivo studies 964:1005 genomic, phylogenetic, and in vivo studies 964:1005 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 28065507 0 44 theme GH70 80:83 arg1 Enzymes 85:91 GH70 Enzymes 80:91 GH70 Enzymes from α-Amylases in Oral Bacteria 80:124 Crystal Structure of 4,6-α-Glucanotransferase Supports Diet-Driven Evolution of GH70 Enzymes from α-Amylases in Oral Bacteria. 28065507 5 45 theme sucrose 1086:1092 arg1 intake 1094:1099 sucrose intake 1086:1099 sucrose intake 1086:1099 Further supported by genomic, phylogenetic, and in vivo studies, we propose that dietary changes involving starch (and starch derivatives) and sucrose intake were critical factors during the evolution of 4,6-α-GTs and glucansucrases from α-amylases, allowing oral bacteria to produce extracellular polymers that contribute to biofilm formation from different substrates. 25113421 10 0 gly glycopeptides 1330:1342 arg2 glycopeptides 1330:1342 intact N-linked glycopeptides 1314:1342 intact N-linked glycopeptides from MS/MS spectra 1314:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 10 0 gly glycopeptides 1330:1342 arg1 spectra 1355:1361 MS/MS spectra 1349:1361 MS/MS spectra 1349:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 12 1 theme GlycoMaster 1530:1540 arg1 DB 1542:1543 GlycoMaster DB 1530:1543 GlycoMaster DB 1530:1543 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 4 2 theme high 625:628 arg1 sensitivity 630:640 its high sensitivity 621:640 its high sensitivity 621:640 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 5 3 theme spectral 709:716 arg1 data 718:721 MS/MS spectral data 703:721 MS/MS spectral data of glycopeptides 703:738 Two experimental approaches exist to obtain MS/MS spectral data of glycopeptides. 25113421 10 4 theme MS/MS 1349:1353 arg1 spectra 1355:1361 MS/MS spectra 1349:1361 MS/MS spectra 1349:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 13 5 theme peptide 1838:1844 arg1 sequences 1846:1854 possible peptide sequences 1829:1854 possible peptide sequences 1829:1854 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 5 6 theme experimental 663:674 arg1 approaches 676:685 Two experimental approaches 659:685 Two experimental approaches 659:685 Two experimental approaches exist to obtain MS/MS spectral data of glycopeptides. 25113421 6 7 theme peptides 842:849 arg1 spectra 815:821 MS/MS spectra 809:821 MS/MS spectra of the glycans and peptides separately 809:860 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 1 8 theme post-translational 170:187 arg1 modifications 189:201 the most commonly observed post-translational modifications 143:201 the most commonly observed post-translational modifications (PTMs) in eukaryotes 143:222 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 1 8 theme post-translational 170:187 arg1 PTMs 204:207 PTMs 204:207 PTMs 204:207 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 6 9 theme glycans 830:836 arg1 spectra 815:821 MS/MS spectra 809:821 MS/MS spectra of the glycans and peptides separately 809:860 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 12 10 theme ETD 1670:1672 arg1 spectra 1674:1680 ETD spectra 1670:1680 ETD spectra 1670:1680 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 11 11 theme sequence 1411:1418 arg1 database 1420:1427 a protein sequence database 1401:1427 a protein sequence database 1401:1427 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 12 12 used used 1686:1689 arg2 spectra 1674:1680 ETD spectra 1670:1680 ETD spectra 1670:1680 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 10 13 from spectra 1355:1361 arg1 identification 1296:1309 the automated and high-throughput identification 1262:1309 the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra 1262:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 10 13 from spectra 1355:1361 arg1 glycopeptides 1330:1342 intact N-linked glycopeptides 1314:1342 intact N-linked glycopeptides from MS/MS spectra 1314:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 11 14 theme glycan 1435:1440 arg1 database 1452:1459 a glycan structure database 1433:1459 a glycan structure database 1433:1459 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 1 15 from modifications 189:201 arg1 eukaryotes 213:222 eukaryotes 213:222 eukaryotes 213:222 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 12 16 theme HCD 1625:1627 arg1 spectra 1629:1635 HCD spectra 1625:1635 HCD spectra 1625:1635 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 9 17 theme software 1099:1106 arg1 lack 1091:1094 the lack 1087:1094 the lack of software specifically designed for the identification of intact glycopeptides 1087:1175 However, the spectral data cannot be readily analyzed because of the lack of software specifically designed for the identification of intact glycopeptides. 25113421 12 18 used used 1641:1644 arg2 spectra 1629:1635 HCD spectra 1625:1635 HCD spectra 1625:1635 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 13 19 theme GlycoMaster 1760:1770 arg1 DB 1772:1773 GlycoMaster DB 1760:1773 GlycoMaster DB 1760:1773 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 10 20 theme glycopeptides 1330:1342 arg1 identification 1296:1309 the automated and high-throughput identification 1262:1309 the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra 1262:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 0 21 theme N-linked 67:74 arg1 glycopeptides 76:88 N-linked glycopeptides 67:88 N-linked glycopeptides 67:88 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 12 22 theme mixed 1598:1602 arg1 fragmentation 1604:1616 HCD/ETD mixed fragmentation 1590:1616 HCD/ETD mixed fragmentation 1590:1616 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 10 23 theme intact 1314:1319 arg1 glycopeptides 1330:1342 intact N-linked glycopeptides 1314:1342 intact N-linked glycopeptides from MS/MS spectra 1314:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 14 24 from http 1987:1990 arg1 accessible 1973:1982 accessible 1973:1982 accessible 1973:1982 GlycoMaster DB is freely accessible at http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB . 25113421 8 25 theme latter 939:944 arg1 approach 946:953 The latter approach 935:953 The latter approach 935:953 The latter approach has the advantage of retaining the glycosylation site information. 25113421 3 26 theme protein-linked 335:348 arg1 glycans 350:356 protein-linked glycans 335:356 protein-linked glycans involved in numerous biological processes 335:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 7 27 gly glycopeptides 920:932 arg2 glycopeptides 920:932 intact glycopeptides 913:932 intact glycopeptides 913:932 The other approach produces spectra directly from intact glycopeptides. 25113421 10 28 link N-linked 1321:1328 arg1 glycopeptides 1330:1342 intact N-linked glycopeptides 1314:1342 intact N-linked glycopeptides from MS/MS spectra 1314:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 11 29 theme input 1514:1518 arg1 spectrum 1520:1527 each input spectrum 1509:1527 each input spectrum 1509:1527 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 0 30 theme mass 100:103 arg1 spectrometry 105:116 tandem mass spectrometry 93:116 tandem mass spectrometry 93:116 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 13 31 gly glycopeptide 1927:1938 arg2 glycopeptide 1927:1938 the N-linked glycopeptide sequon 1914:1945 the N-linked glycopeptide sequon 1914:1945 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 0 32 gly glycopeptides 76:88 arg2 glycopeptides 76:88 N-linked glycopeptides 67:88 N-linked glycopeptides 67:88 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 9 33 theme glycopeptides 1163:1175 arg1 identification 1138:1151 the identification 1134:1151 the identification of intact glycopeptides 1134:1175 However, the spectral data cannot be readily analyzed because of the lack of software specifically designed for the identification of intact glycopeptides. 25113421 0 34 theme GlycoMaster 0:10 arg1 DB 12:13 GlycoMaster DB 0:13 GlycoMaster DB 0:13 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 3 35 theme numerous 370:377 arg1 processes 390:398 numerous biological processes 370:398 numerous biological processes 370:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 3 36 theme structures 482:491 arg1 identification 421:434 the high-throughput identification 401:434 the high-throughput identification of both glycoproteins and the attached glycan structures 401:491 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 4 37 theme glycoproteomic 586:599 arg1 analysis 601:608 glycoproteomic analysis 586:608 glycoproteomic analysis 586:608 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 13 38 theme only 1728:1731 arg1 spectra 1737:1743 only HCD spectra 1728:1743 only HCD spectra 1728:1743 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 13 39 theme precursor 1895:1903 arg1 mass 1905:1908 the accurate precursor mass 1882:1908 the accurate precursor mass 1882:1908 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 3 40 theme attached 466:473 arg1 structures 482:491 the attached glycan structures 462:491 the attached glycan structures 462:491 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 12 41 theme HCD/ETD 1590:1596 arg1 fragmentation 1604:1616 HCD/ETD mixed fragmentation 1590:1616 HCD/ETD mixed fragmentation 1590:1616 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 8 42 theme glycosylation 990:1002 arg1 information 1009:1019 the glycosylation site information 986:1019 the glycosylation site information 986:1019 The latter approach has the advantage of retaining the glycosylation site information. 25113421 10 43 theme novel 1215:1219 arg1 DB 1248:1249 GlycoMaster DB 1236:1249 GlycoMaster DB 1236:1249 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 10 43 theme novel 1215:1219 arg1 tool 1230:1233 a novel software tool 1213:1233 a novel software tool 1213:1233 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 12 44 theme spectral 1562:1569 arg1 data 1571:1574 mass spectral data 1557:1574 mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences 1557:1720 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 13 45 theme sequences 1846:1854 arg1 list 1821:1824 a list 1819:1824 a list of possible peptide sequences 1819:1854 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 7 46 from glycopeptides 920:932 arg1 spectra 891:897 spectra 891:897 spectra directly from intact glycopeptides 891:932 The other approach produces spectra directly from intact glycopeptides. 25113421 3 47 gly glycoproteins 444:456 arg1 glycoproteins 444:456 glycoproteins 444:456 glycoproteins 444:456 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 10 48 theme GlycoMaster 1236:1246 arg1 DB 1248:1249 GlycoMaster DB 1236:1249 GlycoMaster DB 1236:1249 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 10 48 theme GlycoMaster 1236:1246 arg1 tool 1230:1233 a novel software tool 1213:1233 a novel software tool 1213:1233 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 2 49 gly glycosylated 283:294 arg1 proteins 270:277 more than 50% eukaryotic proteins 245:277 more than 50% eukaryotic proteins 245:277 It is believed that more than 50% eukaryotic proteins are glycosylated. 25113421 10 50 theme automated 1266:1274 arg1 identification 1296:1309 the automated and high-throughput identification 1262:1309 the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra 1262:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 4 51 theme mass 533:536 arg1 spectrometry 538:549 Tandem mass spectrometry 526:549 Tandem mass spectrometry (MS/MS) 526:557 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 4 51 theme mass 533:536 arg1 method 575:580 an effective method 562:580 an effective method for glycoproteomic analysis 562:608 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 4 51 theme mass 533:536 arg1 MS/MS 552:556 MS/MS 552:556 MS/MS 552:556 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 6 52 from glycopeptides 780:792 arg1 glycans 767:773 glycans 767:773 glycans from glycopeptides 767:792 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 11 53 theme peptide 1486:1492 arg1 pair 1478:1481 the best pair 1469:1481 the best pair of peptide and glycan 1469:1503 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 5 54 theme MS/MS 703:707 arg1 data 718:721 MS/MS spectral data 703:721 MS/MS spectral data of glycopeptides 703:738 Two experimental approaches exist to obtain MS/MS spectral data of glycopeptides. 25113421 3 55 link protein-linked 335:348 arg1 glycans 350:356 protein-linked glycans 335:356 protein-linked glycans involved in numerous biological processes 335:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 6 56 dep glycans 830:836 arg1 the 826:828 the 826:828 the 826:828 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 13 57 theme possible 1829:1836 arg1 sequences 1846:1854 possible peptide sequences 1829:1854 possible peptide sequences 1829:1854 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 8 58 contain has 955:957 arg1 approach 946:953 The latter approach 935:953 The latter approach 935:953 The latter approach has the advantage of retaining the glycosylation site information. 25113421 8 58 contain has 955:957 arg2 advantage 963:971 the advantage 959:971 the advantage of retaining the glycosylation site information 959:1019 The latter approach has the advantage of retaining the glycosylation site information. 25113421 2 59 theme eukaryotic 259:268 arg1 proteins 270:277 more than 50% eukaryotic proteins 245:277 more than 50% eukaryotic proteins 245:277 It is believed that more than 50% eukaryotic proteins are glycosylated. 25113421 1 60 theme observed 161:168 arg1 modifications 189:201 the most commonly observed post-translational modifications 143:201 the most commonly observed post-translational modifications (PTMs) in eukaryotes 143:222 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 1 60 theme observed 161:168 arg1 PTMs 204:207 PTMs 204:207 PTMs 204:207 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 10 61 from identification 1296:1309 arg1 spectra 1355:1361 MS/MS spectra 1349:1361 MS/MS spectra 1349:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 9 62 theme spectral 1035:1042 arg1 data 1044:1047 the spectral data 1031:1047 the spectral data 1031:1047 However, the spectral data cannot be readily analyzed because of the lack of software specifically designed for the identification of intact glycopeptides. 25113421 7 63 theme intact 913:918 arg1 glycopeptides 920:932 intact glycopeptides 913:932 intact glycopeptides 913:932 The other approach produces spectra directly from intact glycopeptides. 25113421 1 64 theme modifications 189:201 arg1 modifications 189:201 the most commonly observed post-translational modifications 143:201 the most commonly observed post-translational modifications (PTMs) in eukaryotes 143:222 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 1 64 theme modifications 189:201 arg1 one 136:138 one 136:138 one 136:138 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 1 64 theme modifications 189:201 arg1 PTMs 204:207 PTMs 204:207 PTMs 204:207 Glycosylation is one of the most commonly observed post-translational modifications (PTMs) in eukaryotes. 25113421 11 65 theme protein 1403:1409 arg1 database 1420:1427 a protein sequence database 1401:1427 a protein sequence database 1401:1427 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 13 66 theme N-linked 1918:1925 arg1 sequon 1940:1945 the N-linked glycopeptide sequon 1914:1945 the N-linked glycopeptide sequon 1914:1945 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 14 67 theme GlycoMaster 1948:1958 arg1 DB 1960:1961 GlycoMaster DB 1948:1961 GlycoMaster DB 1948:1961 GlycoMaster DB is freely accessible at http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB . 25113421 5 68 gly glycopeptides 726:738 arg2 glycopeptides 726:738 glycopeptides 726:738 glycopeptides 726:738 Two experimental approaches exist to obtain MS/MS spectral data of glycopeptides. 25113421 11 69 theme structure 1442:1450 arg1 database 1452:1459 a glycan structure database 1433:1459 a glycan structure database 1433:1459 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 14 70 dep http 1987:1990 arg1 8080/GlycoMasterDB 2019:2036 8080/GlycoMasterDB 2019:2036 http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB 1987:2036 GlycoMaster DB is freely accessible at http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB . 25113421 14 70 dep http 1987:1990 arg1 //www-novo.cs.uwaterloo.ca 1992:2017 //www-novo.cs.uwaterloo.ca 1992:2017 http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB 1987:2036 GlycoMaster DB is freely accessible at http://www-novo.cs.uwaterloo.ca:8080/GlycoMasterDB . 25113421 5 71 theme glycopeptides 726:738 arg1 data 718:721 MS/MS spectral data 703:721 MS/MS spectral data of glycopeptides 703:738 Two experimental approaches exist to obtain MS/MS spectral data of glycopeptides. 25113421 6 72 gly glycopeptides 780:792 arg2 glycopeptides 780:792 glycopeptides 780:792 glycopeptides 780:792 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 10 73 theme high-throughput 1280:1294 arg1 identification 1296:1309 the automated and high-throughput identification 1262:1309 the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra 1262:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 2 74 dep % 257:257 arg1 50 255:256 50 255:256 50 255:256 It is believed that more than 50% eukaryotic proteins are glycosylated. 25113421 13 75 theme glycopeptide 1927:1938 arg1 sequon 1940:1945 the N-linked glycopeptide sequon 1914:1945 the N-linked glycopeptide sequon 1914:1945 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 0 76 theme glycopeptides 76:88 arg1 identification 49:62 the automated identification 35:62 the automated identification of N-linked glycopeptides 35:88 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 3 77 theme biological 311:320 arg1 functions 322:330 the biological functions 307:330 the biological functions of protein-linked glycans involved in numerous biological processes 307:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 2 78 dep proteins 270:277 arg1 % 257:257 % 257:257 % 257:257 It is believed that more than 50% eukaryotic proteins are glycosylated. 25113421 10 79 theme N-linked 1321:1328 arg1 glycopeptides 1330:1342 intact N-linked glycopeptides 1314:1342 intact N-linked glycopeptides from MS/MS spectra 1314:1361 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 0 80 theme tandem 93:98 arg1 spectrometry 105:116 tandem mass spectrometry 93:116 tandem mass spectrometry 93:116 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 13 81 link N-linked 1918:1925 arg1 sequon 1940:1945 the N-linked glycopeptide sequon 1914:1945 the N-linked glycopeptide sequon 1914:1945 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 3 82 theme glycans 350:356 arg1 functions 322:330 the biological functions 307:330 the biological functions of protein-linked glycans involved in numerous biological processes 307:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 4 83 theme Tandem 526:531 arg1 spectrometry 538:549 Tandem mass spectrometry 526:549 Tandem mass spectrometry (MS/MS) 526:557 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 4 83 theme Tandem 526:531 arg1 method 575:580 an effective method 562:580 an effective method for glycoproteomic analysis 562:608 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 4 83 theme Tandem 526:531 arg1 MS/MS 552:556 MS/MS 552:556 MS/MS 552:556 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 9 84 theme intact 1156:1161 arg1 glycopeptides 1163:1175 intact glycopeptides 1156:1175 intact glycopeptides 1156:1175 However, the spectral data cannot be readily analyzed because of the lack of software specifically designed for the identification of intact glycopeptides. 25113421 3 85 theme glycoproteins 444:456 arg1 identification 421:434 the high-throughput identification 401:434 the high-throughput identification of both glycoproteins and the attached glycan structures 401:491 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 7 86 theme other 867:871 arg1 approach 873:880 The other approach 863:880 The other approach 863:880 The other approach produces spectra directly from intact glycopeptides. 25113421 3 87 theme biological 379:388 arg1 processes 390:398 numerous biological processes 370:398 numerous biological processes 370:398 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 13 88 theme accurate 1886:1893 arg1 mass 1905:1908 the accurate precursor mass 1882:1908 the accurate precursor mass 1882:1908 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 9 89 gly glycopeptides 1163:1175 arg2 glycopeptides 1163:1175 intact glycopeptides 1156:1175 intact glycopeptides 1156:1175 However, the spectral data cannot be readily analyzed because of the lack of software specifically designed for the identification of intact glycopeptides. 25113421 12 90 theme peptide 1704:1710 arg1 sequences 1712:1720 peptide sequences 1704:1720 peptide sequences 1704:1720 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 13 91 theme HCD 1733:1735 arg1 spectra 1737:1743 only HCD spectra 1728:1743 only HCD spectra 1728:1743 When only HCD spectra are available, GlycoMaster DB can still help to identify the glycans, and a list of possible peptide sequences are reported according to the accurate precursor mass and the N-linked glycopeptide sequon. 25113421 3 92 theme glycan 475:480 arg1 structures 482:491 the attached glycan structures 462:491 the attached glycan structures 462:491 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 0 93 theme automated 39:47 arg1 identification 49:62 the automated identification 35:62 the automated identification of N-linked glycopeptides 35:88 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 10 94 theme software 1221:1228 arg1 DB 1248:1249 GlycoMaster DB 1236:1249 GlycoMaster DB 1236:1249 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 10 94 theme software 1221:1228 arg1 tool 1230:1233 a novel software tool 1213:1233 a novel software tool 1213:1233 To address this need, we developed a novel software tool, GlycoMaster DB, to assist the automated and high-throughput identification of intact N-linked glycopeptides from MS/MS spectra. 25113421 8 95 gly glycosylation 990:1002 arg2 information 1009:1019 the glycosylation site information 986:1019 the glycosylation site information 986:1019 The latter approach has the advantage of retaining the glycosylation site information. 25113421 8 95 gly glycosylation 990:1002 arg2 site 1004:1007 the glycosylation site information 986:1019 the glycosylation site information 986:1019 The latter approach has the advantage of retaining the glycosylation site information. 25113421 0 96 link N-linked 67:74 arg1 glycopeptides 76:88 N-linked glycopeptides 67:88 N-linked glycopeptides 67:88 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 3 97 theme high-throughput 405:419 arg1 identification 421:434 the high-throughput identification 401:434 the high-throughput identification of both glycoproteins and the attached glycan structures 401:491 To reveal the biological functions of protein-linked glycans involved in numerous biological processes, the high-throughput identification of both glycoproteins and the attached glycan structures becomes fundamentally important. 25113421 4 98 theme effective 565:573 arg1 spectrometry 538:549 Tandem mass spectrometry 526:549 Tandem mass spectrometry (MS/MS) 526:557 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 4 98 theme effective 565:573 arg1 method 575:580 an effective method 562:580 an effective method for glycoproteomic analysis 562:608 Tandem mass spectrometry (MS/MS) is an effective method for glycoproteomic analysis because of its high sensitivity and selectivity. 25113421 6 99 theme MS/MS 809:813 arg1 spectra 815:821 MS/MS spectra 809:821 MS/MS spectra of the glycans and peptides separately 809:860 One consists of isolating glycans from glycopeptides and generating MS/MS spectra of the glycans and peptides separately. 25113421 0 100 dep software 16:23 arg1 DB 12:13 GlycoMaster DB 0:13 GlycoMaster DB 0:13 GlycoMaster DB: software to assist the automated identification of N-linked glycopeptides by tandem mass spectrometry. 25113421 8 101 theme site 1004:1007 arg1 information 1009:1019 the glycosylation site information 986:1019 the glycosylation site information 986:1019 The latter approach has the advantage of retaining the glycosylation site information. 25113421 11 102 theme glycan 1498:1503 arg1 pair 1478:1481 the best pair 1469:1481 the best pair of peptide and glycan 1469:1503 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25113421 12 103 theme mass 1557:1560 arg1 data 1571:1574 mass spectral data 1557:1574 mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences 1557:1720 GlycoMaster DB can analyze mass spectral data produced with HCD/ETD mixed fragmentation, where HCD spectra are used to identify glycans and ETD spectra are used to determine peptide sequences. 25113421 11 104 theme best 1473:1476 arg1 pair 1478:1481 the best pair 1469:1481 the best pair of peptide and glycan 1469:1503 The software simultaneously searches a protein sequence database and a glycan structure database to find the best pair of peptide and glycan for each input spectrum. 25016576 7 0 theme envelope 1355:1362 arg1 gp140 1373:1377 gp140 1373:1377 gp140 1373:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 0 theme envelope 1355:1362 arg1 protein 1364:1370 the HIV-1 envelope protein 1345:1370 the HIV-1 envelope protein 1345:1370 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 9 1 dep infection 1643:1651 arg1 addition 1617:1624 addition 1617:1624 addition 1617:1624 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 8 2 theme N-linked 1498:1505 arg1 glycosylation 1507:1519 N-linked glycosylation 1498:1519 N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain 1498:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 5 3 theme N-linked 1091:1098 arg1 carbohydrate 1100:1111 N-linked carbohydrate 1091:1111 N-linked carbohydrate 1091:1111 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 9 4 theme gp120 1750:1754 arg1 domain 1740:1745 the V1/V2 domain 1730:1745 the V1/V2 domain of gp120 1730:1754 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 9 4 theme gp120 1750:1754 arg1 gp120 1750:1754 gp120 1750:1754 gp120 1750:1754 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 3 5 theme only 627:630 arg1 response 639:646 the only immune response 623:646 the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial 623:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 3 5 theme only 627:630 arg1 antibodies 564:573 antibodies 564:573 antibodies to the V1/V2 domain 564:593 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 8 6 theme V1/V2 1570:1574 arg1 domain 1576:1581 the gp120 V1/V2 domain 1560:1581 the gp120 V1/V2 domain 1560:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 5 7 link N-linked 1091:1098 arg1 carbohydrate 1100:1111 N-linked carbohydrate 1091:1111 N-linked carbohydrate 1091:1111 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 7 8 theme mAb 1283:1285 arg1 isolation 1262:1270 the isolation 1258:1270 the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140 1258:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 1 9 theme broadly 174:180 arg1 bN-mAbs 218:224 bN-mAbs 218:224 bN-mAbs 218:224 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 1 9 theme broadly 174:180 arg1 antibodies 206:215 several broadly neutralizing monoclonal antibodies 166:215 several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120 166:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 2 10 theme gp120 472:476 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 10 theme gp120 472:476 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 10 11 theme antibody 1814:1821 arg1 responses 1823:1831 antibody responses 1814:1831 antibody responses to GDEs 1814:1839 Although little is known regarding conditions that favor antibody responses to GDEs, our studies demonstrate that these antibodies can arise from a short-term immunization regimen. 25016576 0 12 theme HIV-1 105:109 arg1 protein 120:126 the HIV-1 envelope protein 101:126 the HIV-1 envelope protein 101:126 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 0 12 theme HIV-1 105:109 arg1 gp120 129:133 gp120 129:133 gp120 129:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 1 13 theme monoclonal 195:204 arg1 bN-mAbs 218:224 bN-mAbs 218:224 bN-mAbs 218:224 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 1 13 theme monoclonal 195:204 arg1 antibodies 206:215 several broadly neutralizing monoclonal antibodies 166:215 several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120 166:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 11 14 theme antibodies 2144:2153 arg1 discovery 2120:2128 the discovery 2116:2128 the discovery of additional antibodies to GDEs 2116:2161 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 0 15 theme protein 120:126 arg1 protein 120:126 the HIV-1 envelope protein 101:126 the HIV-1 envelope protein 101:126 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 0 15 theme protein 120:126 arg1 domain 91:96 the V1/V2 domain 81:96 the V1/V2 domain of the HIV-1 envelope protein, gp120 81:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 8 16 from glycosylation 1507:1519 arg1 domain 1576:1581 the gp120 V1/V2 domain 1560:1581 the gp120 V1/V2 domain 1560:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 17 link N-linked 1498:1505 arg1 glycosylation 1507:1519 N-linked glycosylation 1498:1519 N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain 1498:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 3 18 theme HIV-1 693:697 arg1 trial 707:711 the RV144 HIV-1 vaccine trial 683:711 the RV144 HIV-1 vaccine trial 683:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 5 19 from significant 973:983 arg1 vaccines 992:999 HIV vaccines 988:999 HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate 988:1111 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 3 20 theme important 518:526 arg1 role 528:531 an important role 515:531 an important role 515:531 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 11 21 theme responses 2063:2071 arg1 analysis 2042:2049 further analysis 2034:2049 further analysis of antibody responses to the HIV-1 envelope protein 2034:2101 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 3 22 from protection 669:678 arg1 trial 707:711 the RV144 HIV-1 vaccine trial 683:711 the RV144 HIV-1 vaccine trial 683:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 8 23 gly glycopeptide 1402:1413 arg2 glycopeptide 1402:1413 glycopeptide 1402:1413 glycopeptide 1402:1413 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 2 24 theme V1/V2 455:459 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 24 theme V1/V2 455:459 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 0 25 theme V1/V2 85:89 arg1 protein 120:126 the HIV-1 envelope protein 101:126 the HIV-1 envelope protein 101:126 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 0 25 theme V1/V2 85:89 arg1 domain 91:96 the V1/V2 domain 81:96 the V1/V2 domain of the HIV-1 envelope protein, gp120 81:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 2 26 theme variable 445:452 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 26 theme variable 445:452 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 8 27 theme Epitope 1380:1386 arg1 mapping 1388:1394 Epitope mapping 1380:1394 Epitope mapping using glycopeptide fragments 1380:1423 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 28 from N130 1535:1538 arg1 glycosylation 1507:1519 N-linked glycosylation 1498:1519 N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain 1498:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 5 29 theme envelope 1056:1063 arg1 protein 1065:1071 the envelope protein 1052:1071 the envelope protein 1052:1071 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 9 30 theme recombinant 1672:1682 arg1 proteins 1684:1691 recombinant proteins 1672:1691 recombinant proteins 1672:1691 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 8 31 theme in 1429:1430 arg1 mutagenesis 1438:1448 in vitro mutagenesis 1429:1448 in vitro mutagenesis 1429:1448 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 0 32 theme antibody 33:40 arg1 Characterization 0:15 Characterization 0:15 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 0:134 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 1 33 theme envelope 288:295 arg1 gp120 306:310 the HIV-1 envelope protein, gp120 278:310 gp120 306:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 11 34 theme further 2034:2040 arg1 analysis 2042:2049 further analysis 2034:2049 further analysis of antibody responses to the HIV-1 envelope protein 2034:2101 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 5 35 theme molecular 1034:1042 arg1 mass 1044:1047 the molecular mass 1030:1047 the molecular mass of the envelope protein 1030:1071 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 0 36 theme glycan-dependent 53:68 arg1 epitope 70:76 a novel glycan-dependent epitope 45:76 a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120 45:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 7 37 with immunization 1299:1310 arg1 protein 1364:1370 the HIV-1 envelope protein 1345:1370 the HIV-1 envelope protein 1345:1370 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 37 with immunization 1299:1310 arg1 domain 1335:1340 the extracellular domain 1317:1340 the extracellular domain of the HIV-1 envelope protein, gp140 1317:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 11 38 theme HIV-1 2080:2084 arg1 protein 2095:2101 the HIV-1 envelope protein 2076:2101 the HIV-1 envelope protein 2076:2101 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 3 39 theme V1/V2 582:586 arg1 domain 588:593 the V1/V2 domain 578:593 the V1/V2 domain 578:593 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 8 40 theme asparagine 1524:1533 arg1 N130 1535:1538 asparagine N130 1524:1538 asparagine N130 (HXB2 numbering) 1524:1555 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 40 theme asparagine 1524:1533 arg1 numbering 1546:1554 HXB2 numbering 1541:1554 HXB2 numbering 1541:1554 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 7 41 theme protein 1364:1370 arg1 protein 1364:1370 the HIV-1 envelope protein 1345:1370 the HIV-1 envelope protein 1345:1370 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 41 theme protein 1364:1370 arg1 domain 1335:1340 the extracellular domain 1317:1340 the extracellular domain of the HIV-1 envelope protein, gp140 1317:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 10 42 theme immunization 1916:1927 arg1 regimen 1929:1935 a short-term immunization regimen 1903:1935 a short-term immunization regimen 1903:1935 Although little is known regarding conditions that favor antibody responses to GDEs, our studies demonstrate that these antibodies can arise from a short-term immunization regimen. 25016576 5 43 theme mass 1044:1047 arg1 % 1025:1025 50% 1023:1025 50% of the molecular mass of the envelope protein 1023:1071 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 5 43 theme mass 1044:1047 arg1 mass 1044:1047 the molecular mass 1030:1047 the molecular mass of the envelope protein 1030:1071 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 7 44 theme HIV-1 1349:1353 arg1 gp140 1373:1377 gp140 1373:1377 gp140 1373:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 44 theme HIV-1 1349:1353 arg1 protein 1364:1370 the HIV-1 envelope protein 1345:1370 the HIV-1 envelope protein 1345:1370 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 3 45 theme vaccine 699:705 arg1 trial 707:711 the RV144 HIV-1 vaccine trial 683:711 the RV144 HIV-1 vaccine trial 683:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 9 46 theme V1/V2 1734:1738 arg1 domain 1740:1745 the V1/V2 domain 1730:1745 the V1/V2 domain of gp120 1730:1754 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 9 46 theme V1/V2 1734:1738 arg1 gp120 1750:1754 gp120 1750:1754 gp120 1750:1754 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 8 47 theme antibody 1478:1485 arg1 binding 1462:1468 binding 1462:1468 binding of this antibody 1462:1485 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 2 48 theme HIV-1 339:343 arg1 subjects 354:361 HIV-1 infected subjects 339:361 HIV-1 infected subjects 339:361 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 1 49 theme glycan-dependent 242:257 arg1 epitopes 259:266 glycan-dependent epitopes 242:266 glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120 242:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 1 49 theme glycan-dependent 242:257 arg1 GDEs 269:272 GDEs 269:272 GDEs 269:272 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 7 50 theme extracellular 1321:1333 arg1 protein 1364:1370 the HIV-1 envelope protein 1345:1370 the HIV-1 envelope protein 1345:1370 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 50 theme extracellular 1321:1333 arg1 domain 1335:1340 the extracellular domain 1317:1340 the extracellular domain of the HIV-1 envelope protein, gp140 1317:1377 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 3 51 theme immune 632:637 arg1 response 639:646 the only immune response 623:646 the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial 623:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 3 51 theme immune 632:637 arg1 antibodies 564:573 antibodies 564:573 antibodies to the V1/V2 domain 564:593 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 8 52 theme gp120 1564:1568 arg1 domain 1576:1581 the gp120 V1/V2 domain 1560:1581 the gp120 V1/V2 domain 1560:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 4 53 theme antibodies 738:747 arg1 importance 724:733 the importance 720:733 the importance of antibodies to polymeric glycans 720:768 While the importance of antibodies to polymeric glycans is well established for vaccines targeting bacterial diseases, the importance of antibodies to glycans in vaccines targeting HIV has only recently been recognized. 25016576 3 54 theme V1/V2 483:487 arg1 domain 489:494 The V1/V2 domain 479:494 The V1/V2 domain 479:494 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 1 55 theme several 166:172 arg1 bN-mAbs 218:224 bN-mAbs 218:224 bN-mAbs 218:224 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 1 55 theme several 166:172 arg1 antibodies 206:215 several broadly neutralizing monoclonal antibodies 166:215 several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120 166:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 4 56 theme polymeric 752:760 arg1 glycans 762:768 polymeric glycans 752:768 polymeric glycans 752:768 While the importance of antibodies to polymeric glycans is well established for vaccines targeting bacterial diseases, the importance of antibodies to glycans in vaccines targeting HIV has only recently been recognized. 25016576 0 57 theme envelope 111:118 arg1 protein 120:126 the HIV-1 envelope protein 101:126 the HIV-1 envelope protein 101:126 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 0 57 theme envelope 111:118 arg1 gp120 129:133 gp120 129:133 gp120 129:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 9 58 with immunization 1654:1665 arg1 proteins 1684:1691 recombinant proteins 1672:1691 recombinant proteins 1672:1691 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 1 59 theme neutralizing 182:193 arg1 bN-mAbs 218:224 bN-mAbs 218:224 bN-mAbs 218:224 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 1 59 theme neutralizing 182:193 arg1 antibodies 206:215 several broadly neutralizing monoclonal antibodies 166:215 several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120 166:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 7 60 theme mouse 1277:1281 arg1 mAb 1283:1285 a mouse mAb 1275:1285 a mouse mAb 1275:1285 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 7 60 theme mouse 1277:1281 arg1 4B6 1288:1290 4B6 1288:1290 4B6 1288:1290 In this report, we describe the isolation of a mouse mAb, 4B6, after immunization with the extracellular domain of the HIV-1 envelope protein, gp140. 25016576 11 61 theme additional 2133:2142 arg1 antibodies 2144:2153 additional antibodies 2133:2153 additional antibodies to GDEs 2133:2161 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 4 62 from importance 837:846 arg1 vaccines 876:883 vaccines 876:883 vaccines targeting HIV 876:897 While the importance of antibodies to polymeric glycans is well established for vaccines targeting bacterial diseases, the importance of antibodies to glycans in vaccines targeting HIV has only recently been recognized. 25016576 0 63 from epitope 70:76 arg1 protein 120:126 the HIV-1 envelope protein 101:126 the HIV-1 envelope protein 101:126 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 0 63 from epitope 70:76 arg1 domain 91:96 the V1/V2 domain 81:96 the V1/V2 domain of the HIV-1 envelope protein, gp120 81:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 6 64 theme few 1123:1125 arg1 studies 1127:1133 few studies 1123:1133 few studies 1123:1133 However, few studies have reported antibodies to GDEs in humans or animals immunized with candidate HIV-1 vaccines. 25016576 9 65 theme natural 1629:1635 arg1 infection 1643:1651 natural HIV-1 infection 1629:1651 natural HIV-1 infection 1629:1651 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 6 66 theme candidate 1204:1212 arg1 vaccines 1220:1227 candidate HIV-1 vaccines 1204:1227 candidate HIV-1 vaccines 1204:1227 However, few studies have reported antibodies to GDEs in humans or animals immunized with candidate HIV-1 vaccines. 25016576 8 67 dep in 1429:1430 arg1 vitro 1432:1436 vitro 1432:1436 vitro 1432:1436 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 4 68 theme bacterial 813:821 arg1 diseases 823:830 bacterial diseases 813:830 bacterial diseases 813:830 While the importance of antibodies to polymeric glycans is well established for vaccines targeting bacterial diseases, the importance of antibodies to glycans in vaccines targeting HIV has only recently been recognized. 25016576 2 69 theme first 428:432 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 69 theme first 428:432 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 4 70 theme antibodies 851:860 arg1 importance 837:846 the importance 833:846 the importance of antibodies to glycans in vaccines targeting HIV 833:897 While the importance of antibodies to polymeric glycans is well established for vaccines targeting bacterial diseases, the importance of antibodies to glycans in vaccines targeting HIV has only recently been recognized. 25016576 1 71 theme Recent 136:141 arg1 studies 143:149 Recent studies 136:149 Recent studies 136:149 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 5 72 from vaccines 992:999 arg1 significant 973:983 significant 973:983 significant 973:983 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 9 73 theme HIV-1 1637:1641 arg1 infection 1643:1651 natural HIV-1 infection 1629:1651 natural HIV-1 infection 1629:1651 Our results demonstrate that, in addition to natural HIV-1 infection, immunization with recombinant proteins can elicit antibodies to the GDEs in the V1/V2 domain of gp120. 25016576 2 74 theme second 438:443 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 74 theme second 438:443 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 75 from glycans 413:419 arg1 gp120 472:476 gp120 472:476 gp120 472:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 75 from glycans 413:419 arg1 domain 462:467 the first and second variable (V1/V2) domain 424:467 the first and second variable (V1/V2) domain of gp120 424:476 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 11 76 theme envelope 2086:2093 arg1 protein 2095:2101 the HIV-1 envelope protein 2076:2101 the HIV-1 envelope protein 2076:2101 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 5 77 theme HIV 988:990 arg1 vaccines 992:999 HIV vaccines 988:999 HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate 988:1111 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 0 78 theme monoclonal 22:31 arg1 antibody 33:40 a monoclonal antibody 20:40 a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120 20:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 5 79 theme protein 1065:1071 arg1 mass 1044:1047 the molecular mass 1030:1047 the molecular mass of the envelope protein 1030:1071 Antibodies to GDEs may be particularly significant in HIV vaccines based on gp120, where 50% of the molecular mass of the envelope protein is contributed by N-linked carbohydrate. 25016576 1 80 theme HIV-1 282:286 arg1 gp120 306:310 the HIV-1 envelope protein, gp120 278:310 gp120 306:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 11 81 theme antibody 2054:2061 arg1 responses 2063:2071 antibody responses 2054:2071 antibody responses to the HIV-1 envelope protein 2054:2101 Our results suggest that antibodies to GDEs are more common than previously suspected, and that further analysis of antibody responses to the HIV-1 envelope protein will lead to the discovery of additional antibodies to GDEs. 25016576 3 82 theme conformational 536:549 arg1 masking 551:557 conformational masking 536:557 conformational masking 536:557 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 25016576 0 83 theme novel 47:51 arg1 epitope 70:76 a novel glycan-dependent epitope 45:76 a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120 45:133 Characterization of a monoclonal antibody to a novel glycan-dependent epitope in the V1/V2 domain of the HIV-1 envelope protein, gp120. 25016576 1 84 theme protein 297:303 arg1 gp120 306:310 the HIV-1 envelope protein, gp120 278:310 gp120 306:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 8 85 theme glycopeptide 1402:1413 arg1 fragments 1415:1423 glycopeptide fragments 1402:1423 glycopeptide fragments 1402:1423 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 1 86 from epitopes 259:266 arg1 gp120 306:310 the HIV-1 envelope protein, gp120 278:310 gp120 306:310 Recent studies have described several broadly neutralizing monoclonal antibodies (bN-mAbs) that recognize glycan-dependent epitopes (GDEs) in the HIV-1 envelope protein, gp120. 25016576 2 87 dep PG9 383:385 arg1 e.g. 377:380 e.g. 377:380 e.g. 377:380 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 2 87 dep PG9 383:385 arg1 CH03 400:403 CH03 400:403 CH03 400:403 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 6 88 theme HIV-1 1214:1218 arg1 vaccines 1220:1227 candidate HIV-1 vaccines 1204:1227 candidate HIV-1 vaccines 1204:1227 However, few studies have reported antibodies to GDEs in humans or animals immunized with candidate HIV-1 vaccines. 25016576 8 89 theme HXB2 1541:1544 arg1 N130 1535:1538 asparagine N130 1524:1538 asparagine N130 (HXB2 numbering) 1524:1555 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 89 theme HXB2 1541:1544 arg1 numbering 1546:1554 HXB2 numbering 1541:1554 HXB2 numbering 1541:1554 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 6 90 from GDEs 1163:1166 arg1 animals 1181:1187 animals 1181:1187 animals 1181:1187 However, few studies have reported antibodies to GDEs in humans or animals immunized with candidate HIV-1 vaccines. 25016576 6 90 from GDEs 1163:1166 arg1 humans 1171:1176 humans 1171:1176 humans 1171:1176 However, few studies have reported antibodies to GDEs in humans or animals immunized with candidate HIV-1 vaccines. 25016576 8 91 gly glycosylation 1507:1519 arg1 domain 1576:1581 the gp120 V1/V2 domain 1560:1581 the gp120 V1/V2 domain 1560:1581 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 91 gly glycosylation 1507:1519 arg1 numbering 1546:1554 HXB2 numbering 1541:1554 HXB2 numbering 1541:1554 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 91 gly glycosylation 1507:1519 arg1 N130 1535:1538 asparagine N130 1524:1538 asparagine N130 (HXB2 numbering) 1524:1555 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 91 gly glycosylation 1507:1519 arg2 N130 1535:1538 asparagine N130 1524:1538 asparagine N130 (HXB2 numbering) 1524:1555 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 8 91 gly glycosylation 1507:1519 arg2 numbering 1546:1554 HXB2 numbering 1541:1554 HXB2 numbering 1541:1554 Epitope mapping using glycopeptide fragments and in vitro mutagenesis showed that binding of this antibody depends on N-linked glycosylation at asparagine N130 (HXB2 numbering) in the gp120 V1/V2 domain. 25016576 10 92 theme short-term 1905:1914 arg1 regimen 1929:1935 a short-term immunization regimen 1903:1935 a short-term immunization regimen 1903:1935 Although little is known regarding conditions that favor antibody responses to GDEs, our studies demonstrate that these antibodies can arise from a short-term immunization regimen. 25016576 2 93 theme infected 345:352 arg1 subjects 354:361 HIV-1 infected subjects 339:361 HIV-1 infected subjects 339:361 These were recovered from HIV-1 infected subjects, and several (e.g., PG9, PG16, CH01, CH03) target glycans in the first and second variable (V1/V2) domain of gp120. 25016576 3 94 theme RV144 687:691 arg1 trial 707:711 the RV144 HIV-1 vaccine trial 683:711 the RV144 HIV-1 vaccine trial 683:711 The V1/V2 domain is thought to play an important role in conformational masking, and antibodies to the V1/V2 domain were recently identified as the only immune response that correlated with protection in the RV144 HIV-1 vaccine trial. 24279413 5 0 gly glycopeptides 947:959 arg2 glycopeptides 947:959 glycopeptides 947:959 glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 947:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 6 1 theme target-decoy 1181:1192 arg1 approach 1201:1208 a target-decoy search approach 1179:1208 a target-decoy search approach 1179:1208 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 4 2 with glycopeptides 783:795 arg1 glycans 811:817 N-linked glycans 802:817 N-linked glycans attached to their glycosylation sites 802:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 2 3 theme sugars 244:249 arg1 structure 225:233 the structure 221:233 the structure of these sugars 221:249 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 4 4 link N-linked 802:809 arg1 glycans 811:817 N-linked glycans 802:817 N-linked glycans attached to their glycosylation sites 802:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 7 5 dep pooled 1320:1325 arg1 increase 1339:1346 increase 1339:1346 to increase identification confidence 1336:1372 The power of our method is further enhanced when multiple data sets are pooled together to increase identification confidence. 24279413 3 6 theme inherent 572:579 arg1 complexity 581:590 the inherent complexity 568:590 the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites 568:680 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 8 7 link N-linked 1418:1425 arg1 glycopeptides 1427:1439 103 highly confident N-linked glycopeptides 1397:1439 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins 1397:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 7 8 theme method 1265:1270 arg1 power 1252:1256 The power 1248:1256 The power of our method 1248:1270 The power of our method is further enhanced when multiple data sets are pooled together to increase identification confidence. 24279413 5 9 theme glycopeptides 947:959 arg1 spectra 936:942 tandem mass spectra 924:942 tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 924:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 3 10 gly glycosylation 662:674 arg2 sites 676:680 their corresponding glycosylation sites 642:680 their corresponding glycosylation sites 642:680 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 2 11 theme development 331:341 arg1 indicators 309:318 indicators 309:318 indicators of disease development and progression 309:357 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 4 12 link N-linked 754:761 arg1 glycopeptides 763:775 intact N-linked glycopeptides 747:775 intact N-linked glycopeptides 747:775 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 4 13 gly glycopeptides 783:795 arg2 glycopeptides 783:795 glycopeptides 783:795 glycopeptides with N-linked glycans attached to their glycosylation sites 783:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 5 14 theme tandem 924:929 arg1 spectra 936:942 tandem mass spectra 924:942 tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 924:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 8 15 theme proteomic 1554:1562 arg1 platforms 1564:1572 conventional proteomic platforms 1541:1572 conventional proteomic platforms 1541:1572 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 4 16 theme N-linked 802:809 arg1 glycans 811:817 N-linked glycans 802:817 N-linked glycans attached to their glycosylation sites 802:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 1 17 theme important 107:115 arg1 modification 125:136 an important protein modification 104:136 an important protein modification that involves enzymatic attachment of sugars to amino acid residues 104:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 1 17 theme important 107:115 arg1 Glycosylation 87:99 Glycosylation 87:99 Glycosylation 87:99 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 2 18 theme glycosylation 270:282 arg1 effects 259:265 the effects 255:265 the effects of glycosylation 255:282 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 5 19 theme C-trap 1028:1033 arg1 HCD 1049:1051 HCD 1049:1051 HCD 1049:1051 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 19 theme C-trap 1028:1033 arg1 dissociation 1035:1046 higher-energy C-trap dissociation 1014:1046 higher-energy C-trap dissociation (HCD) 1014:1052 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 4 20 theme N-linked 754:761 arg1 glycopeptides 763:775 intact N-linked glycopeptides 747:775 intact N-linked glycopeptides 747:775 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 0 21 theme complex 70:76 arg1 samples 78:84 complex samples 70:84 complex samples 70:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 3 22 theme glycosylation 662:674 arg1 sites 676:680 their corresponding glycosylation sites 642:680 their corresponding glycosylation sites 642:680 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 5 23 theme dissociation 994:1005 arg1 modes 1110:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 8 24 theme abundant 1612:1619 arg1 proteins 1621:1628 the 7-most abundant proteins 1601:1628 the 7-most abundant proteins 1601:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 7 25 theme data 1306:1309 arg1 sets 1311:1314 multiple data sets 1297:1314 multiple data sets 1297:1314 The power of our method is further enhanced when multiple data sets are pooled together to increase identification confidence. 24279413 1 26 theme enzymatic 152:160 arg1 attachment 162:171 enzymatic attachment 152:171 enzymatic attachment of sugars to amino acid residues 152:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 0 27 from glycopeptides 53:65 arg1 samples 78:84 complex samples 70:84 complex samples 70:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 5 28 theme transfer 1068:1075 arg1 ETD 1091:1093 ETD 1091:1093 ETD 1091:1093 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 28 theme transfer 1068:1075 arg1 dissociation 1077:1088 electron transfer dissociation 1059:1088 electron transfer dissociation (ETD) 1059:1094 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 0 29 theme Computational 0:12 arg1 framework 14:22 Computational framework 0:22 Computational framework for identification of intact glycopeptides in complex samples 0:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 3 30 theme spectrometric 405:417 arg1 data 419:422 mass spectrometric data 400:422 mass spectrometric data 400:422 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 9 31 theme site-specific 1709:1721 arg1 glycosylation 1731:1743 site-specific protein glycosylation 1709:1743 site-specific protein glycosylation 1709:1743 These results indicate that our method is ready to be used for characterizing site-specific protein glycosylation in complex samples. 24279413 3 32 theme developing 490:499 arg1 methods 506:512 developing such methods 490:512 developing such methods for the glycoproteome 490:534 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 1 33 theme protein 117:123 arg1 modification 125:136 an important protein modification 104:136 an important protein modification that involves enzymatic attachment of sugars to amino acid residues 104:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 1 33 theme protein 117:123 arg1 Glycosylation 87:99 Glycosylation 87:99 Glycosylation 87:99 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 6 34 theme rate 1146:1149 arg1 method 1162:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 8 35 theme complex 1498:1504 arg1 samples 1527:1533 complex human serum proteome samples 1498:1533 complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins 1498:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 9 36 theme protein 1723:1729 arg1 glycosylation 1731:1743 site-specific protein glycosylation 1709:1743 site-specific protein glycosylation 1709:1743 These results indicate that our method is ready to be used for characterizing site-specific protein glycosylation in complex samples. 24279413 6 37 theme empirical 1120:1128 arg1 method 1162:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 8 38 theme serum 1512:1516 arg1 samples 1527:1533 complex human serum proteome samples 1498:1533 complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins 1498:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 0 39 theme glycopeptides 53:65 arg1 identification 28:41 identification 28:41 identification of intact glycopeptides in complex samples 28:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 0 40 from identification 28:41 arg1 samples 78:84 complex samples 70:84 complex samples 70:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 4 41 theme proteome 869:876 arg1 samples 878:884 complex proteome samples 861:884 complex proteome samples 861:884 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 8 42 from sites 1449:1453 arg1 glycopeptides 1427:1439 103 highly confident N-linked glycopeptides 1397:1439 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins 1397:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 2 43 dep vital 288:292 arg1 Understanding 207:219 Understanding 207:219 Understanding the structure of these sugars 207:249 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 8 44 theme conventional 1541:1552 arg1 platforms 1564:1572 conventional proteomic platforms 1541:1572 conventional proteomic platforms 1541:1572 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 3 45 from changes 466:472 arg1 glycome 481:487 the glycome 477:487 the glycome 477:487 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 1 46 theme acid 192:195 arg1 residues 197:204 amino acid residues 186:204 amino acid residues 186:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 6 47 theme search 1194:1199 arg1 approach 1201:1208 a target-decoy search approach 1179:1208 a target-decoy search approach 1179:1208 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 9 48 gly glycosylation 1731:1743 arg1 samples 1756:1762 complex samples 1748:1762 complex samples 1748:1762 These results indicate that our method is ready to be used for characterizing site-specific protein glycosylation in complex samples. 24279413 9 49 used used 1685:1688 arg2 method 1663:1668 our method 1659:1668 our method 1659:1668 These results indicate that our method is ready to be used for characterizing site-specific protein glycosylation in complex samples. 24279413 1 50 attach attachment 162:171 arg1 residues 197:204 amino acid residues 186:204 amino acid residues 186:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 1 50 attach attachment 162:171 arg2 sugars 176:181 sugars 176:181 sugars 176:181 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 1 51 theme amino 186:190 arg1 residues 197:204 amino acid residues 186:204 amino acid residues 186:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 4 52 theme computational 707:719 arg1 framework 721:729 a computational framework 705:729 a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites 705:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 2 53 theme disease 323:329 arg1 development 331:341 disease development 323:341 disease development 323:341 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 6 54 theme assigning 1226:1234 arg1 confidence 1236:1245 assigning confidence 1226:1245 assigning confidence 1226:1245 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 5 55 theme mass 931:934 arg1 spectra 936:942 tandem mass spectra 924:942 tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 924:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 0 56 from samples 78:84 arg1 identification 28:41 identification 28:41 identification of intact glycopeptides in complex samples 28:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 4 57 attach attached 819:826 arg2 glycans 811:817 N-linked glycans 802:817 N-linked glycans attached to their glycosylation sites 802:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 4 57 attach attached 819:826 arg1 sites 851:855 their glycosylation sites 831:855 their glycosylation sites 831:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 3 58 theme glycan 619:624 arg1 structures 626:635 glycan structures 619:635 glycan structures 619:635 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 4 59 gly glycopeptides 763:775 arg2 glycopeptides 763:775 intact N-linked glycopeptides 747:775 intact N-linked glycopeptides 747:775 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 9 60 theme complex 1748:1754 arg1 samples 1756:1762 complex samples 1748:1762 complex samples 1748:1762 These results indicate that our method is ready to be used for characterizing site-specific protein glycosylation in complex samples. 24279413 5 61 theme higher-energy 1014:1026 arg1 HCD 1049:1051 HCD 1049:1051 HCD 1049:1051 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 61 theme higher-energy 1014:1026 arg1 dissociation 1035:1046 higher-energy C-trap dissociation 1014:1046 higher-energy C-trap dissociation (HCD) 1014:1052 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 8 62 theme 7-most 1605:1610 arg1 proteins 1621:1628 the 7-most abundant proteins 1601:1628 the 7-most abundant proteins 1601:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 4 63 gly glycosylation 837:849 arg2 sites 851:855 their glycosylation sites 831:855 their glycosylation sites 831:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 3 64 theme corresponding 648:660 arg1 sites 676:680 their corresponding glycosylation sites 642:680 their corresponding glycosylation sites 642:680 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 5 65 theme collision-induced 976:992 arg1 CID 1008:1010 CID 1008:1010 CID 1008:1010 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 65 theme collision-induced 976:992 arg1 dissociation 994:1005 collision-induced dissociation 976:1005 collision-induced dissociation (CID) 976:1011 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 3 66 theme computational 369:381 arg1 methods 383:389 computational methods 369:389 computational methods based on mass spectrometric data 369:422 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 8 67 theme proteins 1621:1628 arg1 depletion 1588:1596 standard depletion 1579:1596 standard depletion of the 7-most abundant proteins 1579:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 4 68 theme intact 747:752 arg1 glycopeptides 763:775 intact N-linked glycopeptides 747:775 intact N-linked glycopeptides 747:775 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 7 69 theme multiple 1297:1304 arg1 sets 1311:1314 multiple data sets 1297:1314 multiple data sets 1297:1314 The power of our method is further enhanced when multiple data sets are pooled together to increase identification confidence. 24279413 1 70 theme sugars 176:181 arg1 attachment 162:171 enzymatic attachment 152:171 enzymatic attachment of sugars to amino acid residues 152:204 Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. 24279413 8 71 theme N-linked 1418:1425 arg1 glycopeptides 1427:1439 103 highly confident N-linked glycopeptides 1397:1439 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins 1397:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 8 72 theme proteome 1518:1525 arg1 samples 1527:1533 complex human serum proteome samples 1498:1533 complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins 1498:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 3 73 theme mass 400:403 arg1 data 419:422 mass spectrometric data 400:422 mass spectrometric data 400:422 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 4 74 theme glycosylation 837:849 arg1 sites 851:855 their glycosylation sites 831:855 their glycosylation sites 831:855 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 5 75 theme electron 1059:1066 arg1 ETD 1091:1093 ETD 1091:1093 ETD 1091:1093 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 75 theme electron 1059:1066 arg1 dissociation 1077:1088 electron transfer dissociation 1059:1088 electron transfer dissociation (ETD) 1059:1094 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 5 76 theme Scoring 887:893 arg1 algorithms 895:904 Scoring algorithms 887:904 Scoring algorithms 887:904 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 8 77 gly glycopeptides 1427:1439 arg2 glycopeptides 1427:1439 103 highly confident N-linked glycopeptides 1397:1439 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins 1397:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 8 77 gly glycopeptides 1427:1439 arg1 sites 1449:1453 53 sites 1446:1453 53 sites across 33 glycoproteins 1446:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 5 78 theme dissociation 1035:1046 arg1 modes 1110:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 4 79 dep glycopeptides 783:795 arg1 i.e. 778:781 i.e. 778:781 i.e. 778:781 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 6 80 theme estimation 1151:1160 arg1 method 1162:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 3 81 theme such 501:504 arg1 methods 506:512 developing such methods 490:512 developing such methods for the glycoproteome 490:534 Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. 24279413 0 82 theme intact 46:51 arg1 glycopeptides 53:65 intact glycopeptides 46:65 intact glycopeptides in complex samples 46:84 Computational framework for identification of intact glycopeptides in complex samples. 24279413 5 83 theme fragmentation 1096:1108 arg1 modes 1110:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 6 84 theme false-discovery 1130:1144 arg1 method 1162:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method 1117:1167 An empirical false-discovery rate estimation method, based on a target-decoy search approach, is derived for assigning confidence. 24279413 8 85 gly glycoproteins 1465:1477 arg1 glycoproteins 1465:1477 33 glycoproteins 1462:1477 33 glycoproteins 1462:1477 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 8 86 theme human 1506:1510 arg1 samples 1527:1533 complex human serum proteome samples 1498:1533 complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins 1498:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 8 87 theme standard 1579:1586 arg1 depletion 1588:1596 standard depletion 1579:1596 standard depletion of the 7-most abundant proteins 1579:1628 Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins. 24279413 5 88 theme dissociation 1077:1088 arg1 modes 1110:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes 976:1114 Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentation modes. 24279413 7 89 theme identification 1348:1361 arg1 confidence 1363:1372 identification confidence 1348:1372 identification confidence 1348:1372 The power of our method is further enhanced when multiple data sets are pooled together to increase identification confidence. 24279413 4 90 theme complex 861:867 arg1 samples 878:884 complex proteome samples 861:884 complex proteome samples 861:884 This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. 24279413 2 91 theme progression 347:357 arg1 indicators 309:318 indicators 309:318 indicators of disease development and progression 309:357 Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. 24279413 0 92 gly glycopeptides 53:65 arg2 glycopeptides 53:65 intact glycopeptides 46:65 intact glycopeptides in complex samples 46:84 Computational framework for identification of intact glycopeptides in complex samples. 25793890 9 0 theme vaccine 1731:1737 arg1 immunogens 1739:1748 vaccine immunogens 1731:1748 vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses 1731:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 10 1 theme vaccine 2006:2012 arg1 trial 2014:2018 the partially successful RV144 HIV vaccine trial 1971:2018 the partially successful RV144 HIV vaccine trial 1971:2018 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 1 2 theme neutralizing 257:268 arg1 antibodies 270:279 neutralizing antibodies 257:279 neutralizing antibodies 257:279 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 25793890 6 3 theme four-stranded 1228:1240 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 10 4 theme CRF01_AE 2035:2042 arg1 immunogens 2044:2053 the two CRF01_AE immunogens 2027:2053 the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) 2027:2096 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 3 5 theme genes 575:579 arg1 Libraries 537:545 Libraries 537:545 Libraries 537:545 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 5 6 dep In 855:856 arg1 vitro 858:862 vitro 858:862 vitro 858:862 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 6 7 gly glycosylation 1048:1060 arg2 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 7 gly glycosylation 1048:1060 arg2 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 7 gly glycosylation 1048:1060 arg2 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 3 8 theme related 558:564 arg1 genes 575:579 closely related envelope genes 550:579 closely related envelope genes 550:579 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 7 9 theme HIV-1 1478:1482 arg1 subjects 1493:1500 HIV-1 infected subjects 1478:1500 HIV-1 infected subjects 1478:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 3 10 theme virus 595:599 arg1 quasi-species 601:613 virus quasi-species 595:613 virus quasi-species 595:613 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 6 11 theme glycosylation 1048:1060 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 11 theme glycosylation 1048:1060 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 11 theme glycosylation 1048:1060 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 12 theme clade 1794:1798 arg1 viruses 1809:1815 clade CRF01_AE viruses 1794:1815 clade CRF01_AE viruses 1794:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 5 13 theme single 899:904 arg1 changes 917:923 single amino acid changes 899:923 single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies 899:1008 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 7 14 theme antibodies 1452:1461 arg1 binding 1428:1434 the binding 1424:1434 the binding of neutralizing antibodies in plasma from HIV-1 infected subjects 1424:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 7 15 theme glycosylation 1386:1398 arg1 site 1400:1403 this glycosylation site 1381:1403 this glycosylation site 1381:1403 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 10 16 theme TH023 2082:2086 arg1 isolates 2088:2095 the A244 and TH023 isolates 2069:2095 isolates 2088:2095 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 0 17 theme gp120 111:115 arg1 gp120 111:115 HIV-1 gp120 105:115 HIV-1 gp120 105:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 0 17 theme gp120 111:115 arg1 domain 95:100 the V1/V2 domain 85:100 the V1/V2 domain of HIV-1 gp120 85:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 5 18 theme acid 912:915 arg1 changes 917:923 single amino acid changes 899:923 single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies 899:1008 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 10 19 theme HIV 2002:2004 arg1 trial 2014:2018 the partially successful RV144 HIV vaccine trial 1971:2018 the partially successful RV144 HIV vaccine trial 1971:2018 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 10 20 theme A244 2073:2076 arg1 isolates 2088:2095 the A244 and TH023 isolates 2069:2095 isolates 2088:2095 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 4 21 theme neutralization 770:783 arg1 variants 809:816 neutralization sensitive and resistant variants 770:816 neutralization sensitive and resistant variants 770:816 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 6 22 theme domain 1248:1253 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 2 23 used used 378:381 arg2 we 375:376 we 375:376 we 375:376 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 0 24 theme A 66:66 arg1 strands 74:80 the A and B strands 62:80 the A and B strands of the V1/V2 domain of HIV-1 gp120 62:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 5 25 from changes 917:923 arg1 individuals 934:944 three individuals 928:944 three individuals 928:944 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 7 26 from subjects 1493:1500 arg1 plasma 1466:1471 plasma 1466:1471 plasma from HIV-1 infected subjects 1466:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 7 26 from subjects 1493:1500 arg1 binding 1428:1434 the binding 1424:1434 the binding of neutralizing antibodies in plasma from HIV-1 infected subjects 1424:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 2 27 theme neutralization 495:508 arg1 sensitivity 510:520 neutralization sensitivity 495:520 neutralization sensitivity 495:520 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 8 28 theme domain 1563:1568 arg1 shorter 1604:1610 shorter 1604:1610 shorter 1604:1610 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 8 28 theme domain 1563:1568 arg1 length 1543:1548 the length 1539:1548 the length of the V1/V2 domain in transmitted founder viruses 1539:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 4 29 theme neutralizing 741:752 arg1 antibodies 754:763 broadly neutralizing antibodies 733:763 broadly neutralizing antibodies 733:763 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 0 30 theme B 72:72 arg1 strands 74:80 the A and B strands 62:80 the A and B strands of the V1/V2 domain of HIV-1 gp120 62:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 6 31 dep sites 1062:1066 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 31 dep sites 1062:1066 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 31 dep sites 1062:1066 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 8 32 theme transmitted 1573:1583 arg1 viruses 1593:1599 transmitted founder viruses 1573:1599 transmitted founder viruses 1573:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 6 33 from N149 1092:1095 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 33 from N149 1092:1095 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 34 theme glycosylation 1885:1897 arg1 sites 1899:1903 these glycosylation sites 1879:1903 these glycosylation sites 1879:1903 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 10 35 contain possessed 2103:2111 arg1 immunogens 2044:2053 the two CRF01_AE immunogens 2027:2053 the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) 2027:2096 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 10 35 contain possessed 2103:2111 arg2 sites 2127:2131 glycosylation sites 2113:2131 glycosylation sites 2113:2131 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 4 36 theme sequence 836:843 arg1 analysis 845:852 sequence analysis 836:852 sequence analysis 836:852 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 6 37 from strand 1214:1219 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 2 38 theme genetic 385:391 arg1 approach 393:400 a genetic approach 383:400 a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance 383:534 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 0 39 theme V1/V2 89:93 arg1 gp120 111:115 HIV-1 gp120 105:115 HIV-1 gp120 105:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 0 39 theme V1/V2 89:93 arg1 domain 95:100 the V1/V2 domain 85:100 the V1/V2 domain of HIV-1 gp120 85:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 3 40 attach derived 582:588 arg1 quasi-species 601:613 virus quasi-species 595:613 virus quasi-species 595:613 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 3 40 attach derived 582:588 arg2 Libraries 537:545 Libraries 537:545 Libraries 537:545 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 6 41 from N136 1076:1079 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 41 from N136 1076:1079 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 42 theme recombinant 1759:1769 arg1 proteins 1780:1787 recombinant envelope proteins 1759:1787 recombinant envelope proteins from clade CRF01_AE viruses 1759:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 6 43 theme hypervariable 1114:1126 arg1 peptide 1139:1145 the hypervariable connecting peptide 1110:1145 the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1110:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 44 from N-terminus 1194:1203 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 8 45 from domain 1563:1568 arg1 viruses 1593:1599 transmitted founder viruses 1573:1599 transmitted founder viruses 1573:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 0 46 dep A 66:66 arg1 the 62:64 the 62:64 the 62:64 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 8 47 theme glycosylation 1632:1644 arg1 sites 1646:1650 fewer glycosylation sites 1626:1650 fewer glycosylation sites 1626:1650 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 6 48 theme proximal 1098:1105 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 48 theme proximal 1098:1105 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 48 theme proximal 1098:1105 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 0 49 theme connecting 35:44 arg1 peptide 46:52 the hypervariable connecting peptide 17:52 the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 17:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 7 50 theme monoclonal 1358:1367 arg1 antibodies 1369:1378 broadly neutralizing monoclonal antibodies 1337:1378 broadly neutralizing monoclonal antibodies 1337:1378 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 2 51 theme envelope 463:470 arg1 protein 472:478 the HIV envelope protein 455:478 the HIV envelope protein 455:478 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 8 52 from viruses 1593:1599 arg1 shorter 1604:1610 shorter 1604:1610 shorter 1604:1610 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 8 52 from viruses 1593:1599 arg1 length 1543:1548 the length 1539:1548 the length of the V1/V2 domain in transmitted founder viruses 1539:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 10 53 theme vaccines 1954:1961 arg1 efficacy 1938:1945 the efficacy 1934:1945 the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149 1934:2148 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 10 54 theme successful 1985:1994 arg1 trial 2014:2018 the partially successful RV144 HIV vaccine trial 1971:2018 the partially successful RV144 HIV vaccine trial 1971:2018 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 6 55 theme A 1177:1177 arg1 strand 1179:1184 the A strand 1173:1184 the A strand 1173:1184 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 56 theme proteins 1860:1867 arg1 inclusion 1838:1846 inclusion 1838:1846 inclusion of envelope proteins that lack these glycosylation sites 1838:1903 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 0 57 theme CRF01_AE 165:172 arg1 viruses 174:180 CRF01_AE viruses 165:180 CRF01_AE viruses 165:180 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 4 58 contain containing 722:731 arg1 plasma 715:720 plasma 715:720 plasma containing broadly neutralizing antibodies 715:763 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 4 58 contain containing 722:731 arg2 antibodies 754:763 broadly neutralizing antibodies 733:763 broadly neutralizing antibodies 733:763 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 3 59 theme envelope 566:573 arg1 genes 575:579 closely related envelope genes 550:579 closely related envelope genes 550:579 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 9 60 gly glycosylation 1885:1897 arg2 sites 1899:1903 these glycosylation sites 1879:1903 these glycosylation sites 1879:1903 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 6 61 theme B 1212:1212 arg1 strand 1214:1219 the B strand 1208:1219 the B strand in the four-stranded V1/V2 domain β-sheet structure 1208:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 62 theme envelope 1771:1778 arg1 proteins 1780:1787 recombinant envelope proteins 1759:1787 recombinant envelope proteins from clade CRF01_AE viruses 1759:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 6 63 theme strand 1214:1219 arg1 N-terminus 1194:1203 the N-terminus 1190:1203 the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1190:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 63 theme strand 1214:1219 arg1 C-terminus 1159:1168 the C-terminus 1155:1168 the C-terminus of the A strand 1155:1184 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 7 64 theme infected 1484:1491 arg1 subjects 1493:1500 HIV-1 infected subjects 1478:1500 HIV-1 infected subjects 1478:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 0 65 theme HIV-1 105:109 arg1 gp120 111:115 HIV-1 gp120 105:115 HIV-1 gp120 105:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 6 66 theme N-linked 1039:1046 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 66 theme N-linked 1039:1046 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 66 theme N-linked 1039:1046 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 67 theme CRF01_AE 1800:1807 arg1 viruses 1809:1815 clade CRF01_AE viruses 1794:1815 clade CRF01_AE viruses 1794:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 6 68 link N-linked 1039:1046 arg1 one 1085:1087 one 1085:1087 one 1085:1087 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 68 link N-linked 1039:1046 arg1 two 1069:1071 two 1069:1071 two 1069:1071 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 68 link N-linked 1039:1046 arg1 sites 1062:1066 N-linked glycosylation sites 1039:1066 N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1039:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 5 69 theme amino 906:910 arg1 changes 917:923 single amino acid changes 899:923 single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies 899:1008 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 7 70 theme neutralizing 1439:1450 arg1 antibodies 1452:1461 neutralizing antibodies 1439:1461 neutralizing antibodies 1439:1461 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 8 71 theme V1/V2 1557:1561 arg1 domain 1563:1568 the V1/V2 domain 1553:1568 the V1/V2 domain in transmitted founder viruses 1553:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 6 72 theme β-sheet 1255:1261 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 9 73 theme envelope 1851:1858 arg1 proteins 1860:1867 envelope proteins 1851:1867 envelope proteins that lack these glycosylation sites 1851:1903 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 2 74 from polymorphisms 438:450 arg1 protein 472:478 the HIV envelope protein 455:478 the HIV envelope protein 455:478 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 6 75 theme V1/V2 1242:1246 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 8 76 contain possesses 1616:1624 arg1 shorter 1604:1610 shorter 1604:1610 shorter 1604:1610 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 8 76 contain possesses 1616:1624 arg2 sites 1646:1650 fewer glycosylation sites 1626:1650 fewer glycosylation sites 1626:1650 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 8 76 contain possesses 1616:1624 arg1 length 1543:1548 the length 1539:1548 the length of the V1/V2 domain in transmitted founder viruses 1539:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 1 77 theme HIV 345:347 arg1 infection 349:357 HIV infection 345:357 HIV infection 345:357 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 25793890 8 78 attach isolated 1672:1679 arg1 infections 1694:1703 chronic infections 1686:1703 chronic infections 1686:1703 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 8 78 attach isolated 1672:1679 arg2 viruses 1664:1670 viruses 1664:1670 viruses isolated from chronic infections 1664:1703 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 9 79 from viruses 1809:1815 arg1 proteins 1780:1787 recombinant envelope proteins 1759:1787 recombinant envelope proteins from clade CRF01_AE viruses 1759:1815 Our results suggest that vaccine immunogens based on recombinant envelope proteins from clade CRF01_AE viruses might be improved by inclusion of envelope proteins that lack these glycosylation sites. 25793890 8 80 theme founder 1585:1591 arg1 viruses 1593:1599 transmitted founder viruses 1573:1599 transmitted founder viruses 1573:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 6 81 from structure 1263:1271 arg1 N-terminus 1194:1203 the N-terminus 1190:1203 the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1190:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 81 from structure 1263:1271 arg1 C-terminus 1159:1168 the C-terminus 1155:1168 the C-terminus of the A strand 1155:1184 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 10 82 attach derived 2056:2062 arg2 immunogens 2044:2053 the two CRF01_AE immunogens 2027:2053 the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) 2027:2096 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 10 82 attach derived 2056:2062 arg1 isolates 2088:2095 the A244 and TH023 isolates 2069:2095 isolates 2088:2095 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 10 83 theme glycosylation 2113:2125 arg1 sites 2127:2131 glycosylation sites 2113:2131 glycosylation sites 2113:2131 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 0 84 theme domain 95:100 arg1 strands 74:80 the A and B strands 62:80 the A and B strands of the V1/V2 domain of HIV-1 gp120 62:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 8 85 gly glycosylation 1632:1644 arg2 sites 1646:1650 fewer glycosylation sites 1626:1650 fewer glycosylation sites 1626:1650 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 7 86 gly glycosylation 1386:1398 arg2 site 1400:1403 this glycosylation site 1381:1403 this glycosylation site 1381:1403 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 7 87 from binding 1428:1434 arg1 plasma 1466:1471 plasma 1466:1471 plasma from HIV-1 infected subjects 1466:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 7 87 from binding 1428:1434 arg1 subjects 1493:1500 HIV-1 infected subjects 1478:1500 HIV-1 infected subjects 1478:1500 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 6 88 theme connecting 1128:1137 arg1 peptide 1139:1145 the hypervariable connecting peptide 1110:1145 the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1110:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 8 89 theme fewer 1626:1630 arg1 sites 1646:1650 fewer glycosylation sites 1626:1650 fewer glycosylation sites 1626:1650 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 4 90 theme sensitive 785:793 arg1 variants 809:816 neutralization sensitive and resistant variants 770:816 neutralization sensitive and resistant variants 770:816 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 0 91 theme hypervariable 21:33 arg1 peptide 46:52 the hypervariable connecting peptide 17:52 the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 17:115 Glycans flanking the hypervariable connecting peptide between the A and B strands of the V1/V2 domain of HIV-1 gp120 confer resistance to antibodies that neutralize CRF01_AE viruses. 25793890 7 92 theme antibodies 1369:1378 arg1 binding 1326:1332 the binding 1322:1332 the binding of broadly neutralizing monoclonal antibodies 1322:1378 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 2 93 theme occurring 428:436 arg1 polymorphisms 438:450 naturally occurring polymorphisms 418:450 naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance 418:534 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 4 94 theme resistant 799:807 arg1 variants 809:816 neutralization sensitive and resistant variants 770:816 neutralization sensitive and resistant variants 770:816 The libraries were screened with plasma containing broadly neutralizing antibodies, and neutralization sensitive and resistant variants were selected for sequence analysis. 25793890 1 95 theme molecular 201:209 arg1 determinants 211:222 the molecular determinants 197:222 the molecular determinants of sensitivity and resistance to neutralizing antibodies 197:279 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 25793890 7 96 theme neutralizing 1345:1356 arg1 antibodies 1369:1378 broadly neutralizing monoclonal antibodies 1337:1378 broadly neutralizing monoclonal antibodies 1337:1378 Although N136 has previously been implicated in the binding of broadly neutralizing monoclonal antibodies, this glycosylation site appears to inhibit the binding of neutralizing antibodies in plasma from HIV-1 infected subjects. 25793890 3 97 theme CRF01_AE 664:671 arg1 viruses 673:679 CRF01_AE viruses 664:679 CRF01_AE viruses 664:679 Libraries of closely related envelope genes, derived from virus quasi-species, were constructed from individuals infected with CRF01_AE viruses. 25793890 8 98 from length 1543:1548 arg1 viruses 1593:1599 transmitted founder viruses 1573:1599 transmitted founder viruses 1573:1599 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 5 99 theme In 855:856 arg1 mutagenesis 864:874 In vitro mutagenesis 855:874 In vitro mutagenesis 855:874 In vitro mutagenesis allowed us to identify single amino acid changes in three individuals that conferred resistance to neutralization by these antibodies. 25793890 1 100 theme vaccines 316:323 arg1 development 301:311 the development 297:311 the development of vaccines designed to prevent HIV infection 297:357 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 25793890 10 101 gly glycosylation 2113:2125 arg2 sites 2127:2131 glycosylation sites 2113:2131 glycosylation sites 2113:2131 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 2 102 theme HIV 459:461 arg1 protein 472:478 the HIV envelope protein 455:478 the HIV envelope protein 455:478 In this study, we used a genetic approach to characterize naturally occurring polymorphisms in the HIV envelope protein that conferred neutralization sensitivity or resistance. 25793890 10 103 theme RV144 1996:2000 arg1 trial 2014:2018 the partially successful RV144 HIV vaccine trial 1971:2018 the partially successful RV144 HIV vaccine trial 1971:2018 This strategy might improve the efficacy of the vaccines used in the partially successful RV144 HIV vaccine trial, where the two CRF01_AE immunogens (derived from the A244 and TH023 isolates) both possessed glycosylation sites at N136 and N149. 25793890 6 104 theme strand 1179:1184 arg1 N-terminus 1194:1203 the N-terminus 1190:1203 the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure 1190:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 6 104 theme strand 1179:1184 arg1 C-terminus 1159:1168 the C-terminus 1155:1168 the C-terminus of the A strand 1155:1184 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 8 105 theme chronic 1686:1692 arg1 infections 1694:1703 chronic infections 1686:1703 chronic infections 1686:1703 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 6 106 from C-terminus 1159:1168 arg1 structure 1263:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 the four-stranded V1/V2 domain β-sheet structure 1224:1271 All three mutations created N-linked glycosylation sites (two at N136 and one at N149) proximal to the hypervariable connecting peptide between the C-terminus of the A strand and the N-terminus of the B strand in the four-stranded V1/V2 domain β-sheet structure. 25793890 1 107 theme sensitivity 227:237 arg1 determinants 211:222 the molecular determinants 197:222 the molecular determinants of sensitivity and resistance to neutralizing antibodies 197:279 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 25793890 8 108 theme Previous 1503:1510 arg1 studies 1512:1518 Previous studies 1503:1518 Previous studies 1503:1518 Previous studies have reported that the length of the V1/V2 domain in transmitted founder viruses is shorter and possesses fewer glycosylation sites compared to viruses isolated from chronic infections. 25793890 1 109 theme resistance 243:252 arg1 determinants 211:222 the molecular determinants 197:222 the molecular determinants of sensitivity and resistance to neutralizing antibodies 197:279 Understanding the molecular determinants of sensitivity and resistance to neutralizing antibodies is critical for the development of vaccines designed to prevent HIV infection. 27506355 7 0 gly glycopeptide 726:737 arg2 glycopeptide 726:737 intact glycopeptide analysis 719:746 intact glycopeptide analysis 719:746 Chem.2010, 82, 10194-10202) for intact glycopeptide analysis has been developed. 27506355 11 1 theme site-specific 1440:1452 arg1 glycosylation 1454:1466 site-specific glycosylation profiling 1440:1476 site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS 1440:1515 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 10 2 theme ions 1267:1270 arg1 information 1225:1235 both m/z and intensity information 1202:1235 both m/z and intensity information of the possible fragmentation ions 1202:1270 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 11 3 theme glycoprotein 1481:1492 arg1 datasets 1494:1501 glycoprotein datasets 1481:1501 glycoprotein datasets 1481:1501 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 13 4 theme maximum 1707:1713 arg1 usability 1715:1723 maximum usability 1707:1723 maximum usability 1707:1723 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 0 5 link N-linked 48:55 arg1 glycopeptides 57:69 intact N-linked glycopeptides 41:69 intact N-linked glycopeptides 41:69 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 2 6 theme intact 361:366 arg1 glycopeptides 368:380 intact glycopeptides 361:380 intact glycopeptides 361:380 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 1 7 theme site-specific 225:237 arg1 glycosylation 247:259 site-specific protein glycosylation 225:259 site-specific protein glycosylation 225:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 5 8 dep 5 589:589 arg1 873-875 592:598 873-875 592:598 873-875 592:598 Methods2008, 5, 873-875), and the glycopeptide fragmentation prediction tool, MassAnalyzer (Zhang et al. 27506355 9 9 theme MassAnalyzer-predicted 1033:1054 arg1 spectra 1056:1062 MassAnalyzer-predicted spectra 1033:1062 MassAnalyzer-predicted spectra 1033:1062 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 0 10 theme predicted 145:153 arg1 searching 172:180 predicted spectral library searching 145:180 predicted spectral library searching 145:180 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 7 11 theme glycopeptide 726:737 arg1 analysis 739:746 intact glycopeptide analysis 719:746 intact glycopeptide analysis 719:746 Chem.2010, 82, 10194-10202) for intact glycopeptide analysis has been developed. 27506355 13 12 theme available 1805:1813 arg1 Pipeline 1780:1787 the Trans-Proteomic Pipeline 1760:1787 the Trans-Proteomic Pipeline (TPP) 1760:1793 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 12 theme available 1805:1813 arg1 suite 1840:1844 a freely available and open-source software suite 1796:1844 a freely available and open-source software suite 1796:1844 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 0 13 theme library 164:170 arg1 searching 172:180 predicted spectral library searching 145:180 predicted spectral library searching 145:180 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 12 14 theme human 1649:1653 arg1 acetylcholinesterase 1655:1674 recombinant human acetylcholinesterase 1637:1674 recombinant human acetylcholinesterase expressed in HEK293 cells 1637:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 13 15 theme open-source 1819:1829 arg1 Pipeline 1780:1787 the Trans-Proteomic Pipeline 1760:1787 the Trans-Proteomic Pipeline (TPP) 1760:1793 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 15 theme open-source 1819:1829 arg1 suite 1840:1844 a freely available and open-source software suite 1796:1844 a freely available and open-source software suite 1796:1844 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 8 16 theme low-energy 856:865 arg1 spectra 904:910 low-energy collision-induced dissociation (CID) spectra 856:910 low-energy collision-induced dissociation (CID) spectra of intact glycopeptides 856:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 2 17 gly glycopeptides 368:380 arg2 glycopeptides 368:380 intact glycopeptides 361:380 intact glycopeptides 361:380 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 10 18 theme improved 1305:1312 arg1 accuracy 1314:1321 improved accuracy 1305:1321 improved accuracy 1305:1321 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 1 19 gly glycopeptide 190:201 arg2 glycopeptide 190:201 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 10 20 theme CID 1117:1119 arg1 spectra 1121:1127 CID spectra 1117:1127 CID spectra 1117:1127 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 8 21 theme dissociation 885:896 arg1 spectra 904:910 low-energy collision-induced dissociation (CID) spectra 856:910 low-energy collision-induced dissociation (CID) spectra of intact glycopeptides 856:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 12 22 theme N-glycosylation 1610:1624 arg1 profile 1626:1632 the site-specific N-glycosylation profile 1592:1632 the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells 1592:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 1 23 theme Intact 183:188 arg1 analysis 206:213 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 1 23 theme Intact 183:188 arg1 frontier 277:284 an important frontier 264:284 an important frontier of proteomics 264:298 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 8 24 theme CID 899:901 arg1 spectra 904:910 low-energy collision-induced dissociation (CID) spectra 856:910 low-energy collision-induced dissociation (CID) spectra of intact glycopeptides 856:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 8 25 theme glycan 825:830 arg1 structure 832:840 the glycan structure 821:840 the glycan structure 821:840 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 0 26 theme collision-induced 85:101 arg1 spectrometry 128:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 5 27 theme prediction 637:646 arg1 tool 648:651 the glycopeptide fragmentation prediction tool 606:651 the glycopeptide fragmentation prediction tool 606:651 Methods2008, 5, 873-875), and the glycopeptide fragmentation prediction tool, MassAnalyzer (Zhang et al. 27506355 1 28 theme MS 203:204 arg1 analysis 206:213 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 1 28 theme MS 203:204 arg1 frontier 277:284 an important frontier 264:284 an important frontier of proteomics 264:298 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 0 29 theme tandem 116:121 arg1 spectrometry 128:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 11 30 gly glycoprotein 1481:1492 arg1 glycoprotein 1481:1492 glycoprotein datasets 1481:1501 glycoprotein datasets 1481:1501 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 8 31 theme intact 915:920 arg1 glycopeptides 922:934 intact glycopeptides 915:934 intact glycopeptides 915:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 11 32 theme human 1379:1383 arg1 glycoprotein 1365:1376 a standard glycoprotein 1354:1376 a standard glycoprotein 1354:1376 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 11 32 theme human 1379:1383 arg1 transferrin 1385:1395 human transferrin 1379:1395 human transferrin 1379:1395 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 10 33 from spectra 1121:1127 arg1 identification 1097:1110 Glycan identification 1090:1110 Glycan identification from CID spectra 1090:1127 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 13 34 theme Pipeline 1780:1787 arg1 SpectraST 1726:1734 SpectraST 1726:1734 SpectraST 1726:1734 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 34 theme Pipeline 1780:1787 arg1 part 1752:1755 part 1752:1755 part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite 1752:1844 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 3 35 theme computational 469:481 arg1 tool 483:486 a new computational tool 463:486 a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al 463:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 0 36 theme Direct 0:5 arg1 determination 24:36 Direct glycan structure determination 0:36 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 0:181 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 5 37 theme glycopeptide 610:621 arg1 tool 648:651 the glycopeptide fragmentation prediction tool 606:651 the glycopeptide fragmentation prediction tool 606:651 Methods2008, 5, 873-875), and the glycopeptide fragmentation prediction tool, MassAnalyzer (Zhang et al. 27506355 3 38 theme library 516:522 arg1 et al 564:568 SpectraST (Lam et al 549:568 SpectraST (Lam et al 549:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 38 theme library 516:522 arg1 tool 543:546 the spectral library building/searching tool 503:546 the spectral library building/searching tool 503:546 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 0 39 theme structure 14:22 arg1 determination 24:36 Direct glycan structure determination 0:36 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 0:181 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 8 40 gly glycopeptides 922:934 arg2 glycopeptides 922:934 intact glycopeptides 915:934 intact glycopeptides 915:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 12 41 theme first 1580:1584 arg1 time 1586:1589 the first time 1576:1589 the first time 1576:1589 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 3 42 dep tool 483:486 arg1 work 457:460 this work 452:460 this work 452:460 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 9 43 gly glycopeptide 962:973 arg2 glycopeptide 962:973 possible glycopeptide sequences 953:983 possible glycopeptide sequences as input 953:992 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 9 44 theme sample-specific 997:1011 arg1 library 1022:1028 a sample-specific spectral library 995:1028 a sample-specific spectral library of MassAnalyzer-predicted spectra 995:1062 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 1 45 dep analysis 206:213 arg1 reveal 218:223 reveal 218:223 to reveal site-specific protein glycosylation 215:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 3 46 theme SpectraST 549:557 arg1 et al 564:568 SpectraST (Lam et al 549:568 SpectraST (Lam et al 549:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 46 theme SpectraST 549:557 arg1 tool 543:546 the spectral library building/searching tool 503:546 the spectral library building/searching tool 503:546 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 0 47 theme N-linked 48:55 arg1 glycopeptides 57:69 intact N-linked glycopeptides 41:69 intact N-linked glycopeptides 41:69 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 10 48 theme fragmentation 1253:1265 arg1 ions 1267:1270 the possible fragmentation ions 1240:1270 the possible fragmentation ions 1240:1270 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 9 49 theme possible 953:960 arg1 sequences 975:983 possible glycopeptide sequences 953:983 possible glycopeptide sequences as input 953:992 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 2 50 theme existing 429:436 arg1 workflows 438:446 existing workflows 429:446 existing workflows 429:446 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 0 51 gly glycopeptides 57:69 arg2 glycopeptides 57:69 intact N-linked glycopeptides 41:69 intact N-linked glycopeptides 41:69 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 13 52 theme software 1831:1838 arg1 Pipeline 1780:1787 the Trans-Proteomic Pipeline 1760:1787 the Trans-Proteomic Pipeline (TPP) 1760:1793 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 52 theme software 1831:1838 arg1 suite 1840:1844 a freely available and open-source software suite 1796:1844 a freely available and open-source software suite 1796:1844 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 9 53 theme sequences 975:983 arg1 list 945:948 a list 943:948 a list of possible glycopeptide sequences as input 943:992 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 3 54 theme Lam 560:562 arg1 et al 564:568 SpectraST (Lam et al 549:568 SpectraST (Lam et al 549:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 54 theme Lam 560:562 arg1 tool 543:546 the spectral library building/searching tool 503:546 the spectral library building/searching tool 503:546 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 11 55 theme datasets 1494:1501 arg1 glycosylation 1454:1466 site-specific glycosylation profiling 1440:1476 site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS 1440:1515 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 2 56 theme MS/MS 344:348 arg1 spectra 350:356 MS/MS spectra 344:356 MS/MS spectra of intact glycopeptides 344:380 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 5 57 gly glycopeptide 610:621 arg2 glycopeptide 610:621 the glycopeptide fragmentation prediction tool 606:651 the glycopeptide fragmentation prediction tool 606:651 Methods2008, 5, 873-875), and the glycopeptide fragmentation prediction tool, MassAnalyzer (Zhang et al. 27506355 9 58 theme spectral 1013:1020 arg1 library 1022:1028 a sample-specific spectral library 995:1028 a sample-specific spectral library of MassAnalyzer-predicted spectra 995:1062 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 2 59 theme glycopeptides 368:380 arg1 spectra 350:356 MS/MS spectra 344:356 MS/MS spectra of intact glycopeptides 344:380 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 11 60 gly glycoprotein 1365:1376 arg1 glycoprotein 1365:1376 a standard glycoprotein 1354:1376 a standard glycoprotein 1354:1376 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 11 60 gly glycoprotein 1365:1376 arg1 transferrin 1385:1395 human transferrin 1379:1395 human transferrin 1379:1395 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 0 61 theme spectral 155:162 arg1 searching 172:180 predicted spectral library searching 145:180 predicted spectral library searching 145:180 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 1 62 theme protein 239:245 arg1 glycosylation 247:259 site-specific protein glycosylation 225:259 site-specific protein glycosylation 225:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 7 63 dep Chem.2010 687:695 arg1 10194-10202 702:712 10194-10202 702:712 10194-10202 702:712 Chem.2010, 82, 10194-10202) for intact glycopeptide analysis has been developed. 27506355 9 64 theme spectra 1056:1062 arg1 library 1022:1028 a sample-specific spectral library 995:1028 a sample-specific spectral library of MassAnalyzer-predicted spectra 995:1062 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 10 65 theme Glycan 1090:1095 arg1 identification 1097:1110 Glycan identification 1090:1110 Glycan identification from CID spectra 1090:1127 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 12 66 gly N-glycosylation 1610:1624 arg1 acetylcholinesterase 1655:1674 recombinant human acetylcholinesterase 1637:1674 recombinant human acetylcholinesterase expressed in HEK293 cells 1637:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 12 67 theme acetylcholinesterase 1655:1674 arg1 profile 1626:1632 the site-specific N-glycosylation profile 1592:1632 the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells 1592:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 7 68 theme intact 719:724 arg1 analysis 739:746 intact glycopeptide analysis 719:746 intact glycopeptide analysis 719:746 Chem.2010, 82, 10194-10202) for intact glycopeptide analysis has been developed. 27506355 13 69 theme data 1853:1856 arg1 analysis 1858:1865 MS data analysis 1850:1865 MS data analysis 1850:1865 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 2 70 theme computational 310:322 arg1 tools 324:328 computational tools 310:328 computational tools for analyzing MS/MS spectra of intact glycopeptides 310:380 However, computational tools for analyzing MS/MS spectra of intact glycopeptides are still limited and not well-integrated into existing workflows. 27506355 0 71 theme low-energy 74:83 arg1 spectrometry 128:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 8 72 theme collision-induced 867:883 arg1 spectra 904:910 low-energy collision-induced dissociation (CID) spectra 856:910 low-energy collision-induced dissociation (CID) spectra of intact glycopeptides 856:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 12 73 theme recombinant 1637:1647 arg1 acetylcholinesterase 1655:1674 recombinant human acetylcholinesterase 1637:1674 recombinant human acetylcholinesterase expressed in HEK293 cells 1637:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 13 74 theme MS 1850:1851 arg1 analysis 1858:1865 MS data analysis 1850:1865 MS data analysis 1850:1865 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 1 75 theme important 267:275 arg1 analysis 206:213 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 1 75 theme important 267:275 arg1 frontier 277:284 an important frontier 264:284 an important frontier of proteomics 264:298 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 0 76 theme dissociation 103:114 arg1 spectrometry 128:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 10 77 theme spectral 1144:1151 arg1 library 1153:1159 spectral library 1144:1159 spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy 1144:1321 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 1 78 theme glycopeptide 190:201 arg1 analysis 206:213 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 1 78 theme glycopeptide 190:201 arg1 frontier 277:284 an important frontier 264:284 an important frontier of proteomics 264:298 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 11 79 theme standard 1356:1363 arg1 glycoprotein 1365:1376 a standard glycoprotein 1354:1376 a standard glycoprotein 1354:1376 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 11 79 theme standard 1356:1363 arg1 transferrin 1385:1395 human transferrin 1379:1395 human transferrin 1379:1395 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 0 80 theme mass 123:126 arg1 spectrometry 128:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 low-energy collision-induced dissociation tandem mass spectrometry 74:139 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 12 81 theme site-specific 1596:1608 arg1 profile 1626:1632 the site-specific N-glycosylation profile 1592:1632 the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells 1592:1700 In addition, we further applied our method to reveal, for the first time, the site-specific N-glycosylation profile of recombinant human acetylcholinesterase expressed in HEK293 cells. 27506355 8 82 theme structure 832:840 arg1 determination 804:816 the determination 800:816 the determination of the glycan structure 800:840 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 13 83 theme Trans-Proteomic 1764:1778 arg1 Pipeline 1780:1787 the Trans-Proteomic Pipeline 1760:1787 the Trans-Proteomic Pipeline (TPP) 1760:1793 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 83 theme Trans-Proteomic 1764:1778 arg1 suite 1840:1844 a freely available and open-source software suite 1796:1844 a freely available and open-source software suite 1796:1844 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 13 83 theme Trans-Proteomic 1764:1778 arg1 TPP 1790:1792 TPP 1790:1792 TPP 1790:1792 For maximum usability, SpectraST is developed as part of the Trans-Proteomic Pipeline (TPP), a freely available and open-source software suite for MS data analysis. 27506355 0 84 theme glycan 7:12 arg1 determination 24:36 Direct glycan structure determination 0:36 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 0:181 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 11 85 gly glycosylation 1454:1466 arg1 datasets 1494:1501 glycoprotein datasets 1481:1501 glycoprotein datasets 1481:1501 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 8 86 theme glycopeptides 922:934 arg1 spectra 904:910 low-energy collision-induced dissociation (CID) spectra 856:910 low-energy collision-induced dissociation (CID) spectra of intact glycopeptides 856:934 Specifically, this tool enables the determination of the glycan structure directly from low-energy collision-induced dissociation (CID) spectra of intact glycopeptides. 27506355 5 87 theme fragmentation 623:635 arg1 tool 648:651 the glycopeptide fragmentation prediction tool 606:651 the glycopeptide fragmentation prediction tool 606:651 Methods2008, 5, 873-875), and the glycopeptide fragmentation prediction tool, MassAnalyzer (Zhang et al. 27506355 3 88 theme building/searching 524:541 arg1 et al 564:568 SpectraST (Lam et al 549:568 SpectraST (Lam et al 549:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 88 theme building/searching 524:541 arg1 tool 543:546 the spectral library building/searching tool 503:546 the spectral library building/searching tool 503:546 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 0 89 theme intact 41:46 arg1 glycopeptides 57:69 intact N-linked glycopeptides 41:69 intact N-linked glycopeptides 41:69 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 3 90 theme new 465:467 arg1 tool 483:486 a new computational tool 463:486 a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al 463:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 91 theme spectral 507:514 arg1 et al 564:568 SpectraST (Lam et al 549:568 SpectraST (Lam et al 549:568 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 3 91 theme spectral 507:514 arg1 tool 543:546 the spectral library building/searching tool 503:546 the spectral library building/searching tool 503:546 In this work, a new computational tool which combines the spectral library building/searching tool, SpectraST (Lam et al. 27506355 0 92 theme glycopeptides 57:69 arg1 determination 24:36 Direct glycan structure determination 0:36 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 0:181 Direct glycan structure determination of intact N-linked glycopeptides by low-energy collision-induced dissociation tandem mass spectrometry and predicted spectral library searching. 27506355 10 93 theme intensity 1215:1223 arg1 information 1225:1235 both m/z and intensity information 1202:1235 both m/z and intensity information of the possible fragmentation ions 1202:1270 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 11 94 theme profiling 1468:1476 arg1 glycosylation 1454:1466 site-specific glycosylation profiling 1440:1476 site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS 1440:1515 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 10 95 theme m/z 1207:1209 arg1 information 1225:1235 both m/z and intensity information 1202:1235 both m/z and intensity information of the possible fragmentation ions 1202:1270 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 11 96 from LC-MS/MS 1508:1515 arg1 glycosylation 1454:1466 site-specific glycosylation profiling 1440:1476 site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS 1440:1515 We validated our method using a standard glycoprotein, human transferrin, and evaluated its potential to be used in site-specific glycosylation profiling of glycoprotein datasets from LC-MS/MS. 27506355 10 97 theme possible 1244:1251 arg1 ions 1267:1270 the possible fragmentation ions 1240:1270 the possible fragmentation ions 1240:1270 Glycan identification from CID spectra is achieved by spectral library searching against this library, in which both m/z and intensity information of the possible fragmentation ions are taken into consideration for improved accuracy. 27506355 9 98 theme glycopeptide 962:973 arg1 sequences 975:983 possible glycopeptide sequences 953:983 possible glycopeptide sequences as input 953:992 Given a list of possible glycopeptide sequences as input, a sample-specific spectral library of MassAnalyzer-predicted spectra is built using SpectraST. 27506355 1 99 theme proteomics 289:298 arg1 analysis 206:213 Intact glycopeptide MS analysis 183:213 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation 183:259 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 27506355 1 99 theme proteomics 289:298 arg1 frontier 277:284 an important frontier 264:284 an important frontier of proteomics 264:298 Intact glycopeptide MS analysis to reveal site-specific protein glycosylation is an important frontier of proteomics. 26306636 4 0 theme N-linked 804:811 arg1 glycans 813:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 1 1 used used 287:290 arg2 Tr7 192:194 Tr7 192:194 Tr7 192:194 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 1 1 used used 287:290 arg2 sialidase 181:189 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 6 2 theme activities 1365:1374 arg1 activities 1365:1374 both the sialidase and trans-sialidase activities 1326:1374 activities 1365:1374 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 6 2 theme activities 1365:1374 arg1 % 1321:1321 more than 90% 1309:1321 more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7 1309:1407 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 2 3 located present 488:494 arg2 N-glycans 473:481 heterogeneous high-mannose type N-glycans 441:481 heterogeneous high-mannose type N-glycans 441:481 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 3 located present 488:494 arg1 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 0 4 theme mass 114:117 arg1 spectrometry 119:130 tandem mass spectrometry 107:130 tandem mass spectrometry 107:130 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 3 5 dep P 755:755 arg1 2 757:757 2 757:757 2 757:757 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 5 dep P 755:755 arg1 GlcNA 759:763 GlcNA 759:763 GlcNA 759:763 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 6 with oligosaccharides 686:701 arg1 compositions 616:627 compositions Man(8-16)GlcNA(c2) 616:646 compositions Man(8-16)GlcNA(c2) 616:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 2 7 from glycosites 531:540 arg1 present 488:494 present 488:494 present 488:494 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 1 8 theme Pichia 210:215 arg1 pastoris 217:224 Pichia pastoris 210:224 Pichia pastoris 210:224 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 3 9 theme Man 746:748 arg1 P 755:755 Man(9-14)P(2)GlcNA(c2) 746:767 Man(9-14)P(2)GlcNA(c2) 746:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 9 theme Man 746:748 arg1 c2 765:766 c2 765:766 c2 765:766 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 5 10 theme interaction 991:1001 arg1 chromatography 1010:1023 hydrophilic interaction liquid chromatography 979:1023 hydrophilic interaction liquid chromatography enrichment of glycopeptides 979:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 3 11 link N-linked 543:550 arg1 glycans 552:558 N-linked glycans 543:558 N-linked glycans in Tr7 543:565 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 6 12 theme deglycosylated 1281:1294 arg1 Tr7 1296:1298 the deglycosylated Tr7 1277:1298 the deglycosylated Tr7 1277:1298 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 3 13 dep compositions 616:627 arg1 c2 644:645 c2 644:645 c2 644:645 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 13 dep compositions 616:627 arg1 GlcNA 638:642 Man(8-16)GlcNA 629:642 compositions Man(8-16)GlcNA(c2) 616:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 4 14 theme pastoris-expressed 906:923 arg1 proteins 925:932 P. pastoris-expressed proteins 903:932 P. pastoris-expressed proteins 903:932 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 3 15 theme P 755:755 arg1 forms 710:714 the forms 706:714 the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2) 706:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 0 16 theme sialidase 61:69 arg1 profiling 19:27 Automated N-glycan profiling 0:27 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase 0:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 5 17 theme automated 1090:1098 arg1 interpretation 1100:1113 automated interpretation 1090:1113 automated interpretation of the mass spectra 1090:1133 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 1 18 gly sialylated 327:336 arg1 oligosaccharides 349:364 sialylated human milk oligosaccharides 327:364 sialylated human milk oligosaccharides 327:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 3 19 from oligosaccharides 594:609 arg1 forms 710:714 the forms 706:714 the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2) 706:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 20 with mono- 658:662 arg1 compositions 616:627 compositions Man(8-16)GlcNA(c2) 616:646 compositions Man(8-16)GlcNA(c2) 616:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 1 21 theme oligosaccharides 349:364 arg1 production 313:322 analytical-scale production 296:322 analytical-scale production of sialylated human milk oligosaccharides 296:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 1 22 theme Trypanosoma 161:171 arg1 rangeli 173:179 mutant Trypanosoma rangeli 154:179 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 6 23 theme sialidase 1335:1343 arg1 activities 1365:1374 both the sialidase and trans-sialidase activities 1326:1374 activities 1365:1374 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 5 24 theme accuracy 1059:1066 arg1 spectrometry 1073:1084 high accuracy mass spectrometry 1054:1084 high accuracy mass spectrometry 1054:1084 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 6 25 gly glycosylated 1392:1403 arg1 Tr7 1405:1407 the glycosylated Tr7 1388:1407 the glycosylated Tr7 1388:1407 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 4 26 contain contained 829:837 arg2 HexNAc 853:858 an additional HexNAc 839:858 an additional HexNAc 839:858 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 4 26 contain contained 829:837 arg1 glycans 813:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 5 27 theme " 1166:1166 arg1 software 1168:1175 in-house developed "MassAI" software 1140:1175 in-house developed "MassAI" software 1140:1175 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 3 28 from mono- 658:662 arg1 forms 710:714 the forms 706:714 the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2) 706:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 2 29 theme Tr7 425:427 arg1 N-glycoprofiling 405:420 Mass spectrometry-based site-specific N-glycoprofiling 367:420 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 367:427 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 0 30 theme Automated 0:8 arg1 profiling 19:27 Automated N-glycan profiling 0:27 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase 0:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 5 31 theme developed 1149:1157 arg1 software 1168:1175 in-house developed "MassAI" software 1140:1175 in-house developed "MassAI" software 1140:1175 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 32 theme MassAI 1160:1165 arg1 software 1168:1175 in-house developed "MassAI" software 1140:1175 in-house developed "MassAI" software 1140:1175 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 2 33 theme N-linked 522:529 arg1 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 34 theme high-mannose 455:466 arg1 N-glycans 473:481 heterogeneous high-mannose type N-glycans 441:481 heterogeneous high-mannose type N-glycans 441:481 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 3 35 with oligosaccharides 594:609 arg1 compositions 616:627 compositions Man(8-16)GlcNA(c2) 616:646 compositions Man(8-16)GlcNA(c2) 616:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 1 36 theme analytical-scale 296:311 arg1 production 313:322 analytical-scale production 296:322 analytical-scale production of sialylated human milk oligosaccharides 296:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 5 37 theme mass 1122:1125 arg1 spectra 1127:1133 the mass spectra 1118:1133 the mass spectra 1118:1133 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 3 38 theme di-phosphorylated 668:684 arg1 oligosaccharides 686:701 di-phosphorylated oligosaccharides 668:701 di-phosphorylated oligosaccharides 668:701 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 39 theme N-linked 543:550 arg1 glycans 552:558 N-linked glycans 543:558 N-linked glycans in Tr7 543:565 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 0 40 theme Trypanosoma 41:51 arg1 rangeli 53:59 mutant Trypanosoma rangeli 34:59 a mutant Trypanosoma rangeli sialidase 32:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 1 41 theme human 338:342 arg1 oligosaccharides 349:364 sialylated human milk oligosaccharides 327:364 sialylated human milk oligosaccharides 327:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 5 42 theme hydrophilic 979:989 arg1 chromatography 1010:1023 hydrophilic interaction liquid chromatography 979:1023 hydrophilic interaction liquid chromatography enrichment of glycopeptides 979:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 43 gly glycopeptides 1039:1051 arg2 glycopeptides 1039:1051 glycopeptides 1039:1051 glycopeptides 1039:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 3 44 theme Man 719:721 arg1 P 728:728 Man(9-15)P(1)GlcNA(c2) 719:740 Man(9-15)P(1)GlcNA(c2) 719:740 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 44 theme Man 719:721 arg1 c2 738:739 c2 738:739 c2 738:739 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 5 45 theme glycan 1204:1209 arg1 analysis 1235:1242 glycan site microheterogeneity analysis 1204:1242 glycan site microheterogeneity analysis 1204:1242 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 6 46 theme glycosylated 1392:1403 arg1 Tr7 1405:1407 the glycosylated Tr7 1388:1407 the glycosylated Tr7 1388:1407 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 5 47 theme method 949:954 arg1 pipeline 956:963 a method pipeline 947:963 a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis 947:1242 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 3 48 theme P 728:728 arg1 forms 710:714 the forms 706:714 the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2) 706:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 2 49 theme spectrometry-based 372:389 arg1 N-glycoprofiling 405:420 Mass spectrometry-based site-specific N-glycoprofiling 367:420 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 367:427 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 1 50 theme trans-sialidase 248:262 arg1 activity 264:271 significant trans-sialidase activity 236:271 significant trans-sialidase activity 236:271 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 4 51 theme phosphorylated 789:802 arg1 glycans 813:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 3 52 from oligosaccharides 686:701 arg1 forms 710:714 the forms 706:714 the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2) 706:767 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 6 53 theme Functional 1245:1254 arg1 analysis 1256:1263 Functional analysis 1245:1263 Functional analysis 1245:1263 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 3 54 theme neutral 586:592 arg1 oligosaccharides 594:609 predominantly neutral oligosaccharides 572:609 predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2) 572:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 1 55 theme rangeli 173:179 arg1 Tr7 192:194 Tr7 192:194 Tr7 192:194 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 1 55 theme rangeli 173:179 arg1 sialidase 181:189 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 5 56 from analysis 1235:1242 arg1 efficient 1191:1199 efficient 1191:1199 efficient 1191:1199 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 6 57 theme trans-sialidase 1349:1363 arg1 activities 1365:1374 both the sialidase and trans-sialidase activities 1326:1374 activities 1365:1374 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 6 58 gly deglycosylated 1281:1294 arg1 Tr7 1296:1298 the deglycosylated Tr7 1277:1298 the deglycosylated Tr7 1277:1298 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 4 59 link N-linked 804:811 arg1 glycans 813:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans 784:819 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 3 60 from glycans 552:558 arg1 Tr7 563:565 Tr7 563:565 Tr7 563:565 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 61 theme Man 629:631 arg1 c2 644:645 c2 644:645 c2 644:645 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 61 theme Man 629:631 arg1 GlcNA 638:642 Man(8-16)GlcNA 629:642 compositions Man(8-16)GlcNA(c2) 616:646 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 5 62 theme glycopeptides 1039:1051 arg1 enrichment 1025:1034 enrichment 1025:1034 enrichment 1025:1034 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 1 63 theme sialylated 327:336 arg1 oligosaccharides 349:364 sialylated human milk oligosaccharides 327:364 sialylated human milk oligosaccharides 327:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 5 64 theme high 1054:1057 arg1 spectrometry 1073:1084 high accuracy mass spectrometry 1054:1084 high accuracy mass spectrometry 1054:1084 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 65 from efficient 1191:1199 arg1 analysis 1235:1242 glycan site microheterogeneity analysis 1204:1242 glycan site microheterogeneity analysis 1204:1242 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 66 theme liquid 1003:1008 arg1 chromatography 1010:1023 hydrophilic interaction liquid chromatography 979:1023 hydrophilic interaction liquid chromatography enrichment of glycopeptides 979:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 4 67 theme additional 842:851 arg1 HexNAc 853:858 an additional HexNAc 839:858 an additional HexNAc 839:858 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 5 68 theme enrichment 1025:1034 arg1 chromatography 1010:1023 hydrophilic interaction liquid chromatography 979:1023 hydrophilic interaction liquid chromatography enrichment of glycopeptides 979:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 4 69 theme P. 903:904 arg1 proteins 925:932 P. pastoris-expressed proteins 903:932 P. pastoris-expressed proteins 903:932 Some phosphorylated N-linked glycans further contained an additional HexNAc, which has not previously been reported in P. pastoris-expressed proteins. 26306636 5 70 theme microheterogeneity 1216:1233 arg1 analysis 1235:1242 glycan site microheterogeneity analysis 1204:1242 glycan site microheterogeneity analysis 1204:1242 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 0 71 theme Pichia 84:89 arg1 pastoris 91:98 Pichia pastoris 84:98 Pichia pastoris 84:98 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 1 72 theme milk 344:347 arg1 oligosaccharides 349:364 sialylated human milk oligosaccharides 327:364 sialylated human milk oligosaccharides 327:364 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 1 73 theme mutant 154:159 arg1 rangeli 173:179 mutant Trypanosoma rangeli 154:179 A mutant Trypanosoma rangeli sialidase 152:189 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 6 74 theme relative 1376:1383 arg1 activities 1365:1374 both the sialidase and trans-sialidase activities 1326:1374 activities 1365:1374 Functional analysis showed that the deglycosylated Tr7 retained more than 90% of both the sialidase and trans-sialidase activities relative to the glycosylated Tr7. 26306636 0 75 theme tandem 107:112 arg1 spectrometry 119:130 tandem mass spectrometry 107:130 tandem mass spectrometry 107:130 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 5 76 theme mass 1068:1071 arg1 spectrometry 1073:1084 high accuracy mass spectrometry 1054:1084 high accuracy mass spectrometry 1054:1084 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 0 77 theme N-glycan 10:17 arg1 profiling 19:27 Automated N-glycan profiling 0:27 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase 0:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 2 78 link N-linked 522:529 arg1 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 79 attach present 488:494 arg2 N-glycans 473:481 heterogeneous high-mannose type N-glycans 441:481 heterogeneous high-mannose type N-glycans 441:481 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 79 attach present 488:494 arg1 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 80 gly glycosites 531:540 arg2 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 81 theme type 468:471 arg1 N-glycans 473:481 heterogeneous high-mannose type N-glycans 441:481 heterogeneous high-mannose type N-glycans 441:481 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 0 82 theme mutant 34:39 arg1 rangeli 53:59 mutant Trypanosoma rangeli 34:59 a mutant Trypanosoma rangeli sialidase 32:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 3 83 dep P 728:728 arg1 1 730:730 1 730:730 1 730:730 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 3 83 dep P 728:728 arg1 GlcNA 732:736 GlcNA 732:736 GlcNA 732:736 N-linked glycans in Tr7 were predominantly neutral oligosaccharides with compositions Man(8-16)GlcNA(c2), but also mono- and di-phosphorylated oligosaccharides in the forms of Man(9-15)P(1)GlcNA(c2) and Man(9-14)P(2)GlcNA(c2), respectively. 26306636 2 84 theme potential 512:520 arg1 glycosites 531:540 all the five potential N-linked glycosites 499:540 all the five potential N-linked glycosites 499:540 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 2 85 theme heterogeneous 441:453 arg1 N-glycans 473:481 heterogeneous high-mannose type N-glycans 441:481 heterogeneous high-mannose type N-glycans 441:481 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 0 86 theme rangeli 53:59 arg1 sialidase 61:69 a mutant Trypanosoma rangeli sialidase 32:69 a mutant Trypanosoma rangeli sialidase 32:69 Automated N-glycan profiling of a mutant Trypanosoma rangeli sialidase expressed in Pichia pastoris, using tandem mass spectrometry and bioinformatics. 26306636 5 87 theme spectra 1127:1133 arg1 chromatography 1010:1023 hydrophilic interaction liquid chromatography 979:1023 hydrophilic interaction liquid chromatography enrichment of glycopeptides 979:1051 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 87 theme spectra 1127:1133 arg1 spectrometry 1073:1084 high accuracy mass spectrometry 1054:1084 high accuracy mass spectrometry 1054:1084 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 87 theme spectra 1127:1133 arg1 interpretation 1100:1113 automated interpretation 1090:1113 automated interpretation of the mass spectra 1090:1133 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 5 88 theme in-house 1140:1147 arg1 software 1168:1175 in-house developed "MassAI" software 1140:1175 in-house developed "MassAI" software 1140:1175 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26306636 1 89 theme significant 236:246 arg1 activity 264:271 significant trans-sialidase activity 236:271 significant trans-sialidase activity 236:271 A mutant Trypanosoma rangeli sialidase, Tr7, expressed in Pichia pastoris, exhibits significant trans-sialidase activity, and has been used for analytical-scale production of sialylated human milk oligosaccharides. 26306636 2 90 theme site-specific 391:403 arg1 N-glycoprofiling 405:420 Mass spectrometry-based site-specific N-glycoprofiling 367:420 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 367:427 Mass spectrometry-based site-specific N-glycoprofiling of Tr7 showed that heterogeneous high-mannose type N-glycans were present at all the five potential N-linked glycosites. 26306636 5 91 theme site 1211:1214 arg1 analysis 1235:1242 glycan site microheterogeneity analysis 1204:1242 glycan site microheterogeneity analysis 1204:1242 We compiled a method pipeline that combined hydrophilic interaction liquid chromatography enrichment of glycopeptides, high accuracy mass spectrometry and automated interpretation of the mass spectra with in-house developed "MassAI" software, which proved efficient in glycan site microheterogeneity analysis. 26208004 0 0 theme Virus 74:78 arg1 Surface 47:53 the Surface 43:53 the Surface of Mature Dengue 2 Virus 43:78 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 1 1 theme antibodies 295:304 arg1 induction 269:277 an induction 266:277 an induction of neutralizing antibodies 266:304 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 7 2 theme N-glycan 1057:1064 arg1 types 1048:1052 Five types 1043:1052 Five types of N-glycan 1043:1064 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 9 3 theme profile 1527:1533 arg1 understanding 1490:1502 a comprehensive understanding 1474:1502 a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1474:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 5 4 theme mature 839:844 arg1 particles 853:861 mature DENV-2 particles 839:861 mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 839:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 4 5 attach attached 654:661 arg1 glycoprotein 668:679 E glycoprotein 666:679 E glycoprotein 666:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 4 5 attach attached 654:661 arg2 N-glycans 635:643 the N-glycans 631:643 the N-glycans that are attached to E glycoprotein 631:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 2 6 gly glycosylation 376:388 arg2 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 2 6 gly glycosylation 376:388 arg2 two 353:355 two 353:355 two 353:355 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 4 7 theme N-glycans 635:643 arg1 structures 617:626 the precise structures 605:626 the precise structures of the N-glycans that are attached to E glycoprotein 605:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 4 8 gly glycoprotein 668:679 arg1 glycoprotein 668:679 E glycoprotein 666:679 E glycoprotein 666:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 0 9 from Characterization 0:15 arg1 Surface 47:53 the Surface 43:53 the Surface of Mature Dengue 2 Virus 43:78 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 8 10 theme recognition 1345:1355 arg1 CRD 1365:1367 CRD 1365:1367 CRD 1365:1367 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 10 theme recognition 1345:1355 arg1 DC-SIGN 1373:1379 DC-SIGN 1373:1379 DC-SIGN 1373:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 10 theme recognition 1345:1355 arg1 domain 1357:1362 the carbohydrate recognition domain 1328:1362 the carbohydrate recognition domain (CRD) of DC-SIGN 1328:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 7 11 theme major 1242:1246 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 11 theme major 1242:1246 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 11 theme major 1242:1246 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 5 12 theme integrated 896:905 arg1 method 907:912 an integrated method 893:912 an integrated method that used both lectin microarray and MALDI-TOF-MS 893:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 5 13 from glycans 828:834 arg1 particles 853:861 mature DENV-2 particles 839:861 mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 839:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 7 14 theme high 1152:1155 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 14 theme high 1152:1155 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 14 theme high 1152:1155 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 5 15 theme insect 876:881 arg1 cells 883:887 insect cells 876:887 insect cells 876:887 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 7 16 theme N-linked 1170:1177 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 16 theme N-linked 1170:1177 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 16 theme N-linked 1170:1177 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 2 17 gly glycoprotein 336:347 arg1 glycoprotein 336:347 DENV E glycoprotein 329:347 DENV E glycoprotein 329:347 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 2 18 theme N-linked 367:374 arg1 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 5 19 theme protein 811:817 arg1 glycans 828:834 envelope protein N-linked glycans 802:834 envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 802:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 6 20 theme N-glycans 1022:1030 arg1 heterogeneity 1000:1012 a high heterogeneity 993:1012 a high heterogeneity of DENV N-glycans 993:1030 By combining these methods, a high heterogeneity of DENV N-glycans was found. 26208004 0 21 theme Insect 93:98 arg1 Cells 100:104 Insect Cells 93:104 Insect Cells 93:104 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 5 22 theme DENV-2 846:851 arg1 particles 853:861 mature DENV-2 particles 839:861 mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 839:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 5 23 link N-linked 819:826 arg1 glycans 828:834 envelope protein N-linked glycans 802:834 envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 802:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 9 24 theme whole 1538:1542 arg1 particles 1551:1559 whole DENV-2 particles 1538:1559 whole DENV-2 particles derived from insect cells 1538:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 3 25 theme proper 482:487 arg1 folding 489:495 the proper folding 478:495 the proper folding of the protein 478:510 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 25 theme proper 482:487 arg1 localization 526:537 its cellular localization 513:537 its cellular localization 513:537 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 25 theme proper 482:487 arg1 interactions 544:555 its interactions 540:555 its interactions with receptors and its immunogenicity 540:593 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 5 26 theme envelope 802:809 arg1 protein 811:817 envelope protein 802:817 envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 802:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 7 27 theme sialic 1139:1144 arg1 acid 1146:1149 sialic acid 1139:1149 sialic acid 1139:1149 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 9 28 theme insect 1574:1579 arg1 cells 1581:1585 insect cells 1574:1585 insect cells 1574:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 3 29 with interactions 544:555 arg1 receptors 562:570 receptors 562:570 receptors 562:570 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 29 with interactions 544:555 arg1 immunogenicity 580:593 its immunogenicity 576:593 its immunogenicity 576:593 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 9 30 theme comprehensive 1476:1488 arg1 understanding 1490:1502 a comprehensive understanding 1474:1502 a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1474:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 2 31 theme E 334:334 arg1 glycoprotein 336:347 DENV E glycoprotein 329:347 DENV E glycoprotein 329:347 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 7 32 link N-linked 1170:1177 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 32 link N-linked 1170:1177 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 32 link N-linked 1170:1177 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 9 33 theme DENV-2 1544:1549 arg1 particles 1551:1559 whole DENV-2 particles 1538:1559 whole DENV-2 particles derived from insect cells 1538:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 5 34 from particles 853:861 arg1 structures 788:797 the structures 784:797 the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 784:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 3 35 gly glycoprotein 436:447 arg1 glycoprotein 436:447 E glycoprotein 434:447 E glycoprotein 434:447 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 8 36 theme carbohydrate 1332:1343 arg1 CRD 1365:1367 CRD 1365:1367 CRD 1365:1367 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 36 theme carbohydrate 1332:1343 arg1 DC-SIGN 1373:1379 DC-SIGN 1373:1379 DC-SIGN 1373:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 36 theme carbohydrate 1332:1343 arg1 domain 1357:1362 the carbohydrate recognition domain 1328:1362 the carbohydrate recognition domain (CRD) of DC-SIGN 1328:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 2 37 theme DENV 329:332 arg1 glycoprotein 336:347 DENV E glycoprotein 329:347 DENV E glycoprotein 329:347 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 0 38 theme Structures 29:38 arg1 Characterization 0:15 Characterization 0:15 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus 0:78 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 1 39 theme dengue 242:247 arg1 vaccine 249:255 a dengue vaccine 240:255 a dengue vaccine based on an induction of neutralizing antibodies 240:304 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 9 40 theme N-linked 1511:1518 arg1 profile 1527:1533 the N-linked glycan profile 1507:1533 the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1507:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 2 41 theme glycosylation 376:388 arg1 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 0 42 theme N-Glycan 20:27 arg1 Structures 29:38 N-Glycan Structures 20:38 N-Glycan Structures 20:38 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 1 43 theme vaccine 249:255 arg1 development 225:235 the development 221:235 the development of a dengue vaccine based on an induction of neutralizing antibodies 221:304 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 3 44 theme protein 504:510 arg1 folding 489:495 the proper folding 478:495 the proper folding of the protein 478:510 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 44 theme protein 504:510 arg1 localization 526:537 its cellular localization 513:537 its cellular localization 513:537 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 44 theme protein 504:510 arg1 interactions 544:555 its interactions 540:555 its interactions with receptors and its immunogenicity 540:593 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 1 45 theme cell 179:182 arg1 receptors 184:192 host cell receptors 174:192 host cell receptors 174:192 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 1 46 gly glycoprotein 121:132 arg1 E 135:135 E 135:135 E 135:135 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 1 46 gly glycoprotein 121:132 arg1 glycoprotein 121:132 DENV envelope glycoprotein 107:132 DENV envelope glycoprotein (E) 107:136 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 7 47 theme N-glycans 1223:1231 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 47 theme N-glycans 1223:1231 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 47 theme N-glycans 1223:1231 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 5 48 used used 919:922 arg2 method 907:912 an integrated method 893:912 an integrated method that used both lectin microarray and MALDI-TOF-MS 893:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 4 49 theme precise 609:615 arg1 structures 617:626 the precise structures 605:626 the precise structures of the N-glycans that are attached to E glycoprotein 605:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 2 50 theme potential 357:365 arg1 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 8 51 theme DC-SIGN 1373:1379 arg1 CRD 1365:1367 CRD 1365:1367 CRD 1365:1367 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 51 theme DC-SIGN 1373:1379 arg1 DC-SIGN 1373:1379 DC-SIGN 1373:1379 DC-SIGN 1373:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 51 theme DC-SIGN 1373:1379 arg1 domain 1357:1362 the carbohydrate recognition domain 1328:1362 the carbohydrate recognition domain (CRD) of DC-SIGN 1328:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 51 theme DC-SIGN 1373:1379 arg1 DENV 1319:1322 DENV 1319:1322 DENV 1319:1322 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 9 52 theme glycan 1520:1525 arg1 profile 1527:1533 the N-linked glycan profile 1507:1533 the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1507:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 7 53 theme mannose-type 1157:1168 arg1 oligosaccharides 1179:1194 high mannose-type N-linked oligosaccharides 1152:1194 high mannose-type N-linked oligosaccharides 1152:1194 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 53 theme mannose-type 1157:1168 arg1 galactosylation 1204:1218 the galactosylation 1200:1218 the galactosylation of N-glycans 1200:1231 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 7 53 theme mannose-type 1157:1168 arg1 structures 1248:1257 the major structures 1238:1257 the major structures that were found 1238:1273 Five types of N-glycan were identified on DENV-2, including mannose, GalNAc, GlcNAc, fucose and sialic acid; high mannose-type N-linked oligosaccharides and the galactosylation of N-glycans were the major structures that were found. 26208004 9 54 link N-linked 1511:1518 arg1 profile 1527:1533 the N-linked glycan profile 1507:1533 the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1507:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 2 55 contain has 349:351 arg1 glycoprotein 336:347 DENV E glycoprotein 329:347 DENV E glycoprotein 329:347 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 2 55 contain has 349:351 arg2 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 3 56 theme cellular 517:524 arg1 folding 489:495 the proper folding 478:495 the proper folding of the protein 478:510 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 3 56 theme cellular 517:524 arg1 localization 526:537 its cellular localization 513:537 its cellular localization 513:537 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 5 57 from structures 788:797 arg1 particles 853:861 mature DENV-2 particles 839:861 mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 839:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 3 58 theme glycoprotein 436:447 arg1 N-glycans 421:429 The N-glycans 417:429 The N-glycans of E glycoprotein 417:447 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 5 59 attach derived 863:869 arg1 cells 883:887 insect cells 876:887 insect cells 876:887 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 5 59 attach derived 863:869 arg2 particles 853:861 mature DENV-2 particles 839:861 mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 839:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 9 60 attach derived 1561:1567 arg1 cells 1581:1585 insect cells 1574:1585 insect cells 1574:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 9 60 attach derived 1561:1567 arg2 particles 1551:1559 whole DENV-2 particles 1538:1559 whole DENV-2 particles derived from insect cells 1538:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 1 61 theme DENV 107:110 arg1 E 135:135 E 135:135 E 135:135 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 1 61 theme DENV 107:110 arg1 glycoprotein 121:132 DENV envelope glycoprotein 107:132 DENV envelope glycoprotein (E) 107:136 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 6 62 gly heterogeneity 1000:1012 arg1 N-glycans 1022:1030 DENV N-glycans 1017:1030 DENV N-glycans 1017:1030 By combining these methods, a high heterogeneity of DENV N-glycans was found. 26208004 5 63 theme lectin 929:934 arg1 microarray 936:945 lectin microarray 929:945 lectin microarray 929:945 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 4 64 theme DENV 731:734 arg1 E 736:736 DENV E 731:736 DENV E 731:736 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 1 65 theme envelope 112:119 arg1 E 135:135 E 135:135 E 135:135 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 1 65 theme envelope 112:119 arg1 glycoprotein 121:132 DENV envelope glycoprotein 107:132 DENV envelope glycoprotein (E) 107:136 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 5 66 theme N-linked 819:826 arg1 glycans 828:834 envelope protein N-linked glycans 802:834 envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 802:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 4 67 theme crystal 710:716 arg1 structure 718:726 the crystal structure 706:726 the crystal structure of DENV E 706:736 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 8 68 theme computational 1399:1411 arg1 experiments 1421:1431 computational docking experiments 1399:1431 computational docking experiments 1399:1431 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 6 69 theme high 995:998 arg1 heterogeneity 1000:1012 a high heterogeneity 993:1012 a high heterogeneity of DENV N-glycans 993:1030 By combining these methods, a high heterogeneity of DENV N-glycans was found. 26208004 8 70 theme docking 1413:1419 arg1 experiments 1421:1431 computational docking experiments 1399:1431 computational docking experiments 1399:1431 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 3 71 theme E 434:434 arg1 glycoprotein 436:447 E glycoprotein 434:447 E glycoprotein 434:447 The N-glycans of E glycoprotein have been shown to influence the proper folding of the protein, its cellular localization, its interactions with receptors and its immunogenicity. 26208004 4 72 theme E 666:666 arg1 glycoprotein 668:679 E glycoprotein 666:679 E glycoprotein 666:679 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 0 73 theme Dengue 65:70 arg1 Virus 74:78 Mature Dengue 2 Virus 58:78 Mature Dengue 2 Virus 58:78 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 2 74 link N-linked 367:374 arg1 sites 390:394 two potential N-linked glycosylation sites 353:394 two potential N-linked glycosylation sites 353:394 It is well known that DENV E glycoprotein has two potential N-linked glycosylation sites at Asn67 and Asn153. 26208004 9 75 theme particles 1551:1559 arg1 profile 1527:1533 the N-linked glycan profile 1507:1533 the N-linked glycan profile of whole DENV-2 particles derived from insect cells 1507:1585 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 1 76 theme host 174:177 arg1 receptors 184:192 host cell receptors 174:192 host cell receptors 174:192 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 5 77 theme glycans 828:834 arg1 structures 788:797 the structures 784:797 the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS 784:962 This study characterized the structures of envelope protein N-linked glycans on mature DENV-2 particles derived from insect cells via an integrated method that used both lectin microarray and MALDI-TOF-MS. 26208004 6 78 theme DENV 1017:1020 arg1 N-glycans 1022:1030 DENV N-glycans 1017:1030 DENV N-glycans 1017:1030 By combining these methods, a high heterogeneity of DENV N-glycans was found. 26208004 1 79 theme main 205:208 arg1 target 210:215 the main target 201:215 the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies 201:304 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 0 80 theme Mature 58:63 arg1 Virus 74:78 Mature Dengue 2 Virus 58:78 Mature Dengue 2 Virus 58:78 Characterization of N-Glycan Structures on the Surface of Mature Dengue 2 Virus Derived from Insect Cells. 26208004 1 81 theme neutralizing 282:293 arg1 antibodies 295:304 neutralizing antibodies 282:304 neutralizing antibodies 282:304 DENV envelope glycoprotein (E) is responsible for interacting with host cell receptors and is the main target for the development of a dengue vaccine based on an induction of neutralizing antibodies. 26208004 4 82 theme E 736:736 arg1 structure 718:726 the crystal structure 706:726 the crystal structure of DENV E 706:736 However, the precise structures of the N-glycans that are attached to E glycoprotein remain elusive, although the crystal structure of DENV E has been determined. 26208004 9 83 theme first 1442:1446 arg1 time 1448:1451 the first time 1438:1451 the first time 1438:1451 For the first time, this study provides a comprehensive understanding of the N-linked glycan profile of whole DENV-2 particles derived from insect cells. 26208004 8 84 from glycan 1309:1314 arg1 CRD 1365:1367 CRD 1365:1367 CRD 1365:1367 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 84 from glycan 1309:1314 arg1 DC-SIGN 1373:1379 DC-SIGN 1373:1379 DC-SIGN 1373:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 84 from glycan 1309:1314 arg1 domain 1357:1362 the carbohydrate recognition domain 1328:1362 the carbohydrate recognition domain (CRD) of DC-SIGN 1328:1379 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 26208004 8 84 from glycan 1309:1314 arg1 DENV 1319:1322 DENV 1319:1322 DENV 1319:1322 Furthermore, a complex between a glycan on DENV and the carbohydrate recognition domain (CRD) of DC-SIGN was mimicked with computational docking experiments. 24931470 5 0 theme different 761:769 arg1 effects 771:777 substantially different effects 747:777 substantially different effects 747:777 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 6 1 from changes 1046:1052 arg1 Env 1057:1059 Env 1057:1059 Env 1057:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 5 2 theme V1/V2 840:844 arg1 destabilization 817:831 CD4-induced destabilization 805:831 CD4-induced destabilization of the V1/V2 and V3 crown 805:857 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 4 3 theme gp41 680:683 arg1 subunit 692:698 the gp41 fusion subunit 676:698 the gp41 fusion subunit 676:698 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 6 4 theme molecule-induced 1014:1029 arg1 changes 1046:1052 CD4- and small molecule-induced conformational changes 999:1052 CD4- and small molecule-induced conformational changes in Env 999:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 0 5 from activation 12:21 arg1 trimer 46:51 a soluble HIV-1 Env trimer 26:51 a soluble HIV-1 Env trimer 26:51 CD4-induced activation in a soluble HIV-1 Env trimer. 24931470 6 6 theme small 1008:1012 arg1 changes 1046:1052 CD4- and small molecule-induced conformational changes 999:1052 CD4- and small molecule-induced conformational changes in Env 999:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 3 7 theme hydrogen-deuterium 358:375 arg1 exchange 377:384 hydrogen-deuterium exchange 358:384 hydrogen-deuterium exchange 358:384 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 5 8 contain have 742:745 arg2 effects 771:777 substantially different effects 747:777 substantially different effects 747:777 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 5 8 contain have 742:745 arg1 inhibitors 731:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 6 9 from Env 1057:1059 arg1 pathways 1080:1087 the allosteric pathways 1065:1087 the allosteric pathways that lead to membrane fusion 1065:1116 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 6 9 from Env 1057:1059 arg1 knowledge 986:994 our knowledge 982:994 our knowledge of CD4- and small molecule-induced conformational changes in Env 982:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 4 10 theme sheet 610:614 arg1 elements 616:623 bridging sheet elements 601:623 bridging sheet elements 601:623 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 1 11 gly glycoprotein 71:82 arg1 Env 85:87 Env 85:87 Env 85:87 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 1 11 gly glycoprotein 71:82 arg1 glycoprotein 71:82 HIV envelope glycoprotein 58:82 The HIV envelope glycoprotein (Env) trimer 54:95 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 6 12 theme CD4- 999:1002 arg1 changes 1046:1052 CD4- and small molecule-induced conformational changes 999:1052 CD4- and small molecule-induced conformational changes in Env 999:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 6 13 from knowledge 986:994 arg1 Env 1057:1059 Env 1057:1059 Env 1057:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 3 14 theme Env 495:497 arg1 trimers 499:505 soluble Env trimers 487:505 soluble Env trimers (SOSIP.664) 487:517 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 3 14 theme Env 495:497 arg1 SOSIP.664 508:516 SOSIP.664 508:516 SOSIP.664 508:516 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 5 15 theme site-targeted 717:729 arg1 inhibitors 731:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 5 16 theme gp120/gp41 908:917 arg1 interface 919:927 the gp120/gp41 interface 904:927 the gp120/gp41 interface 904:927 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 6 17 theme membrane 1102:1109 arg1 fusion 1111:1116 membrane fusion 1102:1116 membrane fusion 1102:1116 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 1 18 theme HIV 58:60 arg1 Env 85:87 Env 85:87 Env 85:87 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 1 18 theme HIV 58:60 arg1 glycoprotein 71:82 HIV envelope glycoprotein 58:82 The HIV envelope glycoprotein (Env) trimer 54:95 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 4 19 theme domain 664:669 arg1 much 640:643 much 640:643 much 640:643 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 4 20 theme gp120 652:656 arg1 domain 664:669 the gp120 inner domain 648:669 the gp120 inner domain 648:669 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 4 21 theme bridging 601:608 arg1 elements 616:623 bridging sheet elements 601:623 bridging sheet elements 601:623 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 1 22 theme envelope 62:69 arg1 Env 85:87 Env 85:87 Env 85:87 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 1 22 theme envelope 62:69 arg1 glycoprotein 71:82 HIV envelope glycoprotein 58:82 The HIV envelope glycoprotein (Env) trimer 54:95 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 3 23 theme unliganded 453:462 arg1 composition 543:553 their glycan composition 530:553 their glycan composition 530:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 3 23 theme unliganded 453:462 arg1 forms 478:482 the unliganded and CD4-bound forms 449:482 the unliganded and CD4-bound forms 449:482 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 5 24 theme CD4-induced 805:815 arg1 destabilization 817:831 CD4-induced destabilization 805:831 CD4-induced destabilization of the V1/V2 and V3 crown 805:857 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 1 25 theme glycoprotein 71:82 arg1 trimer 90:95 The HIV envelope glycoprotein (Env) trimer 54:95 The HIV envelope glycoprotein (Env) trimer 54:95 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 1 26 theme viral 172:176 arg1 membranes 191:199 the viral and cellular membranes 168:199 the viral and cellular membranes 168:199 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 0 27 theme CD4-induced 0:10 arg1 activation 12:21 CD4-induced activation 0:21 CD4-induced activation in a soluble HIV-1 Env trimer. 0:52 CD4-induced activation in a soluble HIV-1 Env trimer. 24931470 6 28 from pathways 1080:1087 arg1 Env 1057:1059 Env 1057:1059 Env 1057:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 4 29 theme inner 658:662 arg1 domain 664:669 the gp120 inner domain 648:669 the gp120 inner domain 648:669 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 4 30 theme elements 616:623 arg1 subunit 692:698 the gp41 fusion subunit 676:698 the gp41 fusion subunit 676:698 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 4 30 theme elements 616:623 arg1 reorganization 583:596 the reorganization 579:596 the reorganization of bridging sheet elements 579:623 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 4 31 theme CD4 556:558 arg1 activation 560:569 CD4 activation 556:569 CD4 activation 556:569 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 1 32 theme cellular 182:189 arg1 membranes 191:199 the viral and cellular membranes 168:199 the viral and cellular membranes 168:199 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 3 33 theme oxidative 390:398 arg1 labeling 400:407 oxidative labeling 390:407 oxidative labeling 390:407 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 2 34 theme large-scale 299:309 arg1 rearrangements 311:324 only large-scale rearrangements 294:324 only large-scale rearrangements 294:324 Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. 24931470 1 35 theme membranes 191:199 arg1 fusion 158:163 fusion 158:163 fusion of the viral and cellular membranes 158:199 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 0 36 theme HIV-1 36:40 arg1 trimer 46:51 a soluble HIV-1 Env trimer 26:51 a soluble HIV-1 Env trimer 26:51 CD4-induced activation in a soluble HIV-1 Env trimer. 24931470 5 37 theme V3 850:851 arg1 destabilization 817:831 CD4-induced destabilization 805:831 CD4-induced destabilization of the V1/V2 and V3 crown 805:857 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 0 38 theme soluble 28:34 arg1 trimer 46:51 a soluble HIV-1 Env trimer 26:51 a soluble HIV-1 Env trimer 26:51 CD4-induced activation in a soluble HIV-1 Env trimer. 24931470 3 39 theme precise 424:430 arg1 understanding 432:444 a more precise understanding 417:444 a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition 417:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 6 40 theme allosteric 1069:1078 arg1 pathways 1080:1087 the allosteric pathways 1065:1087 the allosteric pathways that lead to membrane fusion 1065:1116 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 3 41 theme CD4-bound 468:476 arg1 composition 543:553 their glycan composition 530:553 their glycan composition 530:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 3 41 theme CD4-bound 468:476 arg1 forms 478:482 the unliganded and CD4-bound forms 449:482 the unliganded and CD4-bound forms 449:482 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 5 42 theme CD4 705:707 arg1 inhibitors 731:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 3 43 theme trimers 499:505 arg1 composition 543:553 their glycan composition 530:553 their glycan composition 530:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 3 43 theme trimers 499:505 arg1 forms 478:482 the unliganded and CD4-bound forms 449:482 the unliganded and CD4-bound forms 449:482 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 0 44 theme Env 42:44 arg1 trimer 46:51 a soluble HIV-1 Env trimer 26:51 a soluble HIV-1 Env trimer 26:51 CD4-induced activation in a soluble HIV-1 Env trimer. 24931470 2 45 theme Env 202:204 arg1 changes 221:227 Env conformational changes 202:227 Env conformational changes 202:227 Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. 24931470 2 46 theme conformational 206:219 arg1 changes 221:227 Env conformational changes 202:227 Env conformational changes 202:227 Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. 24931470 4 47 theme fusion 685:690 arg1 subunit 692:698 the gp41 fusion subunit 676:698 the gp41 fusion subunit 676:698 CD4 activation induces the reorganization of bridging sheet elements, V1/V2 and V3, much of the gp120 inner domain, and the gp41 fusion subunit. 24931470 6 48 theme structural 934:943 arg1 information 945:955 The structural information 930:955 The structural information presented here 930:970 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 2 49 theme electron 269:276 arg1 microscopy 278:287 low-resolution electron microscopy 254:287 low-resolution electron microscopy 254:287 Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. 24931470 1 50 theme receptor-induced 107:122 arg1 changes 139:145 receptor-induced conformational changes 107:145 receptor-induced conformational changes that drive fusion of the viral and cellular membranes 107:199 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 3 51 theme glycan 536:541 arg1 composition 543:553 their glycan composition 530:553 their glycan composition 530:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 2 52 theme low-resolution 254:267 arg1 microscopy 278:287 low-resolution electron microscopy 254:287 low-resolution electron microscopy 254:287 Env conformational changes have been observed using low-resolution electron microscopy, but only large-scale rearrangements have been visible. 24931470 1 53 theme conformational 124:137 arg1 changes 139:145 receptor-induced conformational changes 107:145 receptor-induced conformational changes that drive fusion of the viral and cellular membranes 107:199 The HIV envelope glycoprotein (Env) trimer undergoes receptor-induced conformational changes that drive fusion of the viral and cellular membranes. 24931470 3 54 theme soluble 487:493 arg1 trimers 499:505 soluble Env trimers 487:505 soluble Env trimers (SOSIP.664) 487:517 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 3 54 theme soluble 487:493 arg1 SOSIP.664 508:516 SOSIP.664 508:516 SOSIP.664 508:516 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 5 55 dep V1/V2 840:844 arg1 crown 853:857 crown 853:857 crown 853:857 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 5 55 dep V1/V2 840:844 arg1 the 836:838 the 836:838 the 836:838 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 24931470 3 56 theme forms 478:482 arg1 understanding 432:444 a more precise understanding 417:444 a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition 417:553 Here, we use hydrogen-deuterium exchange and oxidative labeling to gain a more precise understanding of the unliganded and CD4-bound forms of soluble Env trimers (SOSIP.664), including their glycan composition. 24931470 6 57 theme changes 1046:1052 arg1 pathways 1080:1087 the allosteric pathways 1065:1087 the allosteric pathways that lead to membrane fusion 1065:1116 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 6 57 theme changes 1046:1052 arg1 knowledge 986:994 our knowledge 982:994 our knowledge of CD4- and small molecule-induced conformational changes in Env 982:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 6 58 theme conformational 1031:1044 arg1 changes 1046:1052 CD4- and small molecule-induced conformational changes 999:1052 CD4- and small molecule-induced conformational changes in Env 999:1059 The structural information presented here increases our knowledge of CD4- and small molecule-induced conformational changes in Env and the allosteric pathways that lead to membrane fusion. 24931470 5 59 theme binding 709:715 arg1 inhibitors 731:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors 701:740 Two CD4 binding site-targeted inhibitors have substantially different effects: NBD-556 partially mimics CD4-induced destabilization of the V1/V2 and V3 crown, whereas BMS-806 only affects regions around the gp120/gp41 interface. 26850169 6 0 theme amino 1044:1048 arg1 substitutions 1055:1067 amino acid substitutions 1044:1067 amino acid substitutions 1044:1067 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 10 1 theme GASDALIE 1473:1480 arg1 Fc 1482:1483 GASDALIE Fc 1473:1483 GASDALIE Fc alone 1473:1489 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 3 2 with macrophages 470:480 arg1 FcγRIIa 555:561 the activating receptor FcγRIIa 531:561 the activating receptor FcγRIIa 531:561 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 1 3 theme Fc 253:254 arg1 receptors 256:264 activating Fc receptors 242:264 activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa) 242:291 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 3 theme Fc 253:254 arg1 FcγRIIIa 283:290 FcγRIIIa 283:290 FcγRIIIa 283:290 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 10 4 theme wild-type 1506:1514 arg1 structures 1519:1528 wild-type Fc structures 1506:1528 wild-type Fc structures 1506:1528 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 7 5 theme GASDALIE 1070:1077 arg1 mutant 1091:1096 an Fc mutant 1085:1096 an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc 1085:1233 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 7 5 theme GASDALIE 1070:1077 arg1 Fc 1079:1080 GASDALIE Fc 1070:1080 GASDALIE Fc 1070:1080 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 10 6 theme increased 1682:1690 arg1 affinity 1692:1699 the increased affinity 1678:1699 the increased affinity of GASDALIE Fc for FcγRIIIa 1678:1727 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 4 7 theme Fc 633:634 arg1 affinity 617:624 The affinity 613:624 The affinity of IgG Fc for binding activating Fc receptors 613:670 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 9 8 theme crystal 1386:1392 arg1 structure 1394:1402 the crystal structure 1382:1402 the crystal structure of GASDALIE Fc alone 1382:1423 We also determined the crystal structure of GASDALIE Fc alone and bound to FcγRIIIa. 26850169 10 9 from interactions 1564:1575 arg1 interface 1605:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 10 10 theme Fc 1713:1714 arg1 affinity 1692:1699 the increased affinity 1678:1699 the increased affinity of GASDALIE Fc for FcγRIIIa 1678:1727 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 1 11 theme antibody-dependent 170:187 arg1 cytotoxicity 203:214 antibody-dependent cell-mediated cytotoxicity 170:214 antibody-dependent cell-mediated cytotoxicity (ADCC) 170:221 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 11 theme antibody-dependent 170:187 arg1 ADCC 217:220 ADCC 217:220 ADCC 217:220 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 3 12 theme activating 535:544 arg1 FcγRIIa 555:561 the activating receptor FcγRIIa 531:561 the activating receptor FcγRIIa 531:561 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 9 13 theme Fc 1416:1417 arg1 structure 1394:1402 the crystal structure 1382:1402 the crystal structure of GASDALIE Fc alone 1382:1423 We also determined the crystal structure of GASDALIE Fc alone and bound to FcγRIIIa. 26850169 5 14 theme Fc 806:807 arg1 regions 809:815 Fc regions 806:815 Fc regions with afucosylated glycans 806:841 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 4 15 theme IgG 683:685 arg1 subclass 687:694 IgG subclass 683:694 IgG subclass 683:694 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 10 16 theme increased 1540:1548 arg1 interactions 1564:1575 increased electrostatic interactions 1540:1575 increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface 1540:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 1 17 theme Immunoglobulin 106:119 arg1 G 121:121 Immunoglobulin G 106:121 Immunoglobulin G (IgG) 106:127 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 17 theme Immunoglobulin 106:119 arg1 IgG 124:126 IgG 124:126 IgG 124:126 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 8 18 theme altered 1251:1257 arg1 functions 1259:1267 its altered functions 1247:1267 its altered functions 1247:1267 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 5 19 theme afucosylated 898:909 arg1 Fcs 911:913 afucosylated Fcs 898:913 afucosylated Fcs 898:913 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 10 20 theme overall 1452:1458 arg1 structure 1460:1468 The overall structure 1448:1468 The overall structure of GASDALIE Fc alone 1448:1489 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 10 20 theme overall 1452:1458 arg1 similar 1495:1501 similar 1495:1501 similar 1495:1501 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 6 21 theme Enhanced 968:975 arg1 responses 994:1002 Enhanced pro-inflammatory responses 968:1002 Enhanced pro-inflammatory responses 968:1002 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 4 22 theme Fc 659:660 arg1 receptors 662:670 activating Fc receptors 648:670 activating Fc receptors 648:670 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 7 23 theme effector 1183:1190 arg1 functions 1192:1200 increased effector functions 1173:1200 increased effector functions 1173:1200 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 4 24 from asparagine 760:769 arg1 domain 785:790 the Fc CH2 domain 774:790 the Fc CH2 domain 774:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 4 25 theme N-linked 719:726 arg1 glycans 728:734 N-linked glycans 719:734 N-linked glycans attached to a conserved asparagine in the Fc CH2 domain 719:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 0 26 theme receptor 70:77 arg1 FcγRIIIa 79:86 the activating Fc receptor FcγRIIIa 52:86 the activating Fc receptor FcγRIIIa 52:86 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 1 27 theme inflammatory 139:150 arg1 cytotoxicity 203:214 antibody-dependent cell-mediated cytotoxicity 170:214 antibody-dependent cell-mediated cytotoxicity (ADCC) 170:221 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 27 theme inflammatory 139:150 arg1 responses 152:160 inflammatory responses 139:160 inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) 139:221 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 5 28 theme enhanced 923:930 arg1 activity 937:944 enhanced ADCC activity 923:944 enhanced ADCC activity 923:944 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 8 29 theme GASDALIE 1300:1307 arg1 affinities 1286:1295 the affinities 1282:1295 the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs 1282:1360 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 7 30 theme higher 1140:1145 arg1 affinity 1147:1154 a higher affinity 1138:1154 a higher affinity for FcγRIIIa and increased effector functions 1138:1200 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 6 31 theme Fc 1028:1029 arg1 regions 1031:1037 Fc regions 1028:1037 Fc regions with amino acid substitutions 1028:1067 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 0 32 theme Fc 67:68 arg1 receptor 70:77 the activating Fc receptor 52:77 the activating Fc receptor FcγRIIIa 52:86 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 8 33 theme wild-type 1313:1321 arg1 Fc 1323:1324 wild-type Fc 1313:1324 wild-type Fc 1313:1324 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 10 34 theme GASDALIE 1584:1591 arg1 interface 1605:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 0 35 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of GASDALIE Fc 0:41 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 2 36 theme killer 407:412 arg1 cells 414:418 natural killer cells 399:418 natural killer cells 399:418 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 5 37 dep bind 843:846 arg1 exhibit 915:921 exhibit 915:921 bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro 843:965 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 4 38 theme CH2 781:783 arg1 domain 785:790 the Fc CH2 domain 774:790 the Fc CH2 domain 774:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 0 39 theme Fc 40:41 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of GASDALIE Fc 0:41 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 3 40 with leukocytes 500:509 arg1 FcγRIIa 555:561 the activating receptor FcγRIIa 531:561 the activating receptor FcγRIIa 531:561 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 6 41 with regions 1031:1037 arg1 substitutions 1055:1067 amino acid substitutions 1044:1067 amino acid substitutions 1044:1067 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 0 42 theme activating 56:65 arg1 receptor 70:77 the activating Fc receptor 52:77 the activating Fc receptor FcγRIIIa 52:86 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 10 43 theme FcγR 1638:1641 arg1 structures 1643:1652 FcγR structures 1638:1652 FcγR structures 1638:1652 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 7 44 theme Fc 1088:1089 arg1 mutant 1091:1096 an Fc mutant 1085:1096 an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc 1085:1233 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 7 44 theme Fc 1088:1089 arg1 Fc 1079:1080 GASDALIE Fc 1070:1080 GASDALIE Fc 1070:1080 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 7 44 theme Fc 1088:1089 arg1 G236A/S239D/A330L/I332E 1099:1121 G236A/S239D/A330L/I332E 1099:1121 G236A/S239D/A330L/I332E 1099:1121 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 5 45 with regions 809:815 arg1 glycans 835:841 afucosylated glycans 822:841 afucosylated glycans 822:841 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 4 46 theme Fc 778:779 arg1 domain 785:790 the Fc CH2 domain 774:790 the Fc CH2 domain 774:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 7 47 theme wild-type 1222:1230 arg1 Fc 1232:1233 wild-type Fc 1222:1233 wild-type Fc 1222:1233 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 10 48 theme FcγRIIIa 1596:1603 arg1 interface 1605:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 2 49 theme immune 342:347 arg1 cells 414:418 natural killer cells 399:418 natural killer cells 399:418 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 2 49 theme immune 342:347 arg1 macrophages 365:375 macrophages 365:375 macrophages 365:375 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 2 49 theme immune 342:347 arg1 cells 349:353 immune cells 342:353 immune cells including macrophages, dendritic cells, and natural killer cells 342:418 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 2 49 theme immune 342:347 arg1 cells 388:392 dendritic cells 378:392 dendritic cells 378:392 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 6 50 theme acid 1050:1053 arg1 substitutions 1055:1067 amino acid substitutions 1044:1067 amino acid substitutions 1044:1067 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 10 51 theme Fc 1482:1483 arg1 structure 1460:1468 The overall structure 1448:1468 The overall structure of GASDALIE Fc alone 1448:1489 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 10 51 theme Fc 1482:1483 arg1 similar 1495:1501 similar 1495:1501 similar 1495:1501 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 1 52 theme activating 242:251 arg1 receptors 256:264 activating Fc receptors 242:264 activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa) 242:291 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 52 theme activating 242:251 arg1 FcγRIIIa 283:290 FcγRIIIa 283:290 FcγRIIIa 283:290 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 10 53 theme Fc 1516:1517 arg1 structures 1519:1528 wild-type Fc structures 1506:1528 wild-type Fc structures 1506:1528 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 5 54 gly afucosylated 822:833 arg1 glycans 835:841 afucosylated glycans 822:841 afucosylated glycans 822:841 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 3 55 theme myeloid 492:498 arg1 leukocytes 500:509 other myeloid leukocytes 486:509 other myeloid leukocytes 486:509 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 4 56 theme IgG 629:631 arg1 Fc 633:634 IgG Fc 629:634 IgG Fc 629:634 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 3 57 theme receptor 546:553 arg1 FcγRIIa 555:561 the activating receptor FcγRIIa 531:561 the activating receptor FcγRIIa 531:561 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 5 58 theme afucosylated 822:833 arg1 glycans 835:841 afucosylated glycans 822:841 afucosylated glycans 822:841 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 4 59 attach attached 736:743 arg2 glycans 728:734 N-linked glycans 719:734 N-linked glycans attached to a conserved asparagine in the Fc CH2 domain 719:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 4 59 attach attached 736:743 arg1 asparagine 760:769 a conserved asparagine 748:769 a conserved asparagine in the Fc CH2 domain 748:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 10 60 theme GASDALIE 1704:1711 arg1 Fc 1713:1714 GASDALIE Fc 1704:1714 GASDALIE Fc 1704:1714 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 1 61 theme cell-mediated 189:201 arg1 cytotoxicity 203:214 antibody-dependent cell-mediated cytotoxicity 170:214 antibody-dependent cell-mediated cytotoxicity (ADCC) 170:221 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 1 61 theme cell-mediated 189:201 arg1 ADCC 217:220 ADCC 217:220 ADCC 217:220 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 5 62 gly fucosylated 878:888 arg1 Fc 890:891 fucosylated Fc 878:891 fucosylated Fc 878:891 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 10 63 theme electrostatic 1550:1562 arg1 interactions 1564:1575 increased electrostatic interactions 1540:1575 increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface 1540:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 3 64 theme inhibitory 424:433 arg1 FcγRIIb 445:451 FcγRIIb 445:451 FcγRIIb 445:451 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 3 64 theme inhibitory 424:433 arg1 receptor 435:442 An inhibitory receptor 421:442 An inhibitory receptor 421:442 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 3 65 theme cell 596:599 arg1 activation 601:610 cell activation 596:610 cell activation 596:610 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 26850169 1 66 theme G 121:121 arg1 region 96:101 The Fc region 89:101 The Fc region of Immunoglobulin G (IgG) 89:127 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 6 67 theme pro-inflammatory 977:992 arg1 responses 994:1002 Enhanced pro-inflammatory responses 968:1002 Enhanced pro-inflammatory responses 968:1002 Enhanced pro-inflammatory responses have also been seen for Fc regions with amino acid substitutions. 26850169 7 68 theme increased 1173:1181 arg1 functions 1192:1200 increased effector functions 1173:1200 increased effector functions 1173:1200 GASDALIE Fc is an Fc mutant (G236A/S239D/A330L/I332E) that exhibits a higher affinity for FcγRIIIa and increased effector functions in vivo compared to wild-type Fc. 26850169 9 69 theme GASDALIE 1407:1414 arg1 Fc 1416:1417 GASDALIE Fc 1407:1417 GASDALIE Fc alone 1407:1423 We also determined the crystal structure of GASDALIE Fc alone and bound to FcγRIIIa. 26850169 4 70 theme activating 648:657 arg1 receptors 662:670 activating Fc receptors 648:670 activating Fc receptors 648:670 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 4 71 theme glycans 728:734 arg1 subclass 687:694 IgG subclass 683:694 IgG subclass 683:694 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 4 71 theme glycans 728:734 arg1 composition 704:714 the composition 700:714 the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain 700:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 5 72 theme ADCC 932:935 arg1 activity 937:944 enhanced ADCC activity 923:944 enhanced ADCC activity 923:944 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 5 73 theme fucosylated 878:888 arg1 Fc 890:891 fucosylated Fc 878:891 fucosylated Fc 878:891 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 5 74 gly afucosylated 898:909 arg1 Fcs 911:913 afucosylated Fcs 898:913 afucosylated Fcs 898:913 For example, Fc regions with afucosylated glycans bind more tightly to FcγRIIIa than fucosylated Fc, and afucosylated Fcs exhibit enhanced ADCC activity in vivo and in vitro. 26850169 10 75 theme Fc 1593:1594 arg1 interface 1605:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 the GASDALIE Fc:FcγRIIIa interface 1580:1613 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 2 76 theme cells 349:353 arg1 surface 331:337 the surface 327:337 the surface of immune cells including macrophages, dendritic cells, and natural killer cells 327:418 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 0 77 theme GASDALIE 31:38 arg1 Fc 40:41 GASDALIE Fc 31:41 GASDALIE Fc 31:41 Structural characterization of GASDALIE Fc bound to the activating Fc receptor FcγRIIIa. 26850169 1 78 theme Fc 93:94 arg1 region 96:101 The Fc region 89:101 The Fc region of Immunoglobulin G (IgG) 89:127 The Fc region of Immunoglobulin G (IgG) initiates inflammatory responses such as antibody-dependent cell-mediated cytotoxicity (ADCC) through binding to activating Fc receptors (FcγRI, FcγRIIa, FcγRIIIa). 26850169 2 79 theme natural 399:405 arg1 cells 414:418 natural killer cells 399:418 natural killer cells 399:418 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 8 80 theme Fc 1323:1324 arg1 affinities 1286:1295 the affinities 1282:1295 the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs 1282:1360 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 8 81 theme activating 1330:1339 arg1 FcγRs 1356:1360 activating and inhibitory FcγRs 1330:1360 activating and inhibitory FcγRs 1330:1360 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 2 82 theme dendritic 378:386 arg1 cells 388:392 dendritic cells 378:392 dendritic cells 378:392 These receptors are expressed on the surface of immune cells including macrophages, dendritic cells, and natural killer cells. 26850169 8 83 theme inhibitory 1345:1354 arg1 FcγRs 1356:1360 activating and inhibitory FcγRs 1330:1360 activating and inhibitory FcγRs 1330:1360 To explore its altered functions, we compared the affinities of GASDALIE and wild-type Fc for activating and inhibitory FcγRs. 26850169 4 84 theme conserved 750:758 arg1 asparagine 760:769 a conserved asparagine 748:769 a conserved asparagine in the Fc CH2 domain 748:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 10 85 theme other 1629:1633 arg1 Fc 1635:1636 other Fc 1629:1636 other Fc 1629:1636 The overall structure of GASDALIE Fc alone was similar to wild-type Fc structures, however, increased electrostatic interactions in the GASDALIE Fc:FcγRIIIa interface compared with other Fc:FcγR structures suggest a mechanism for the increased affinity of GASDALIE Fc for FcγRIIIa. 26850169 4 86 link N-linked 719:726 arg1 glycans 728:734 N-linked glycans 719:734 N-linked glycans attached to a conserved asparagine in the Fc CH2 domain 719:790 The affinity of IgG Fc for binding activating Fc receptors depends on IgG subclass and the composition of N-linked glycans attached to a conserved asparagine in the Fc CH2 domain. 26850169 3 87 theme other 486:490 arg1 leukocytes 500:509 other myeloid leukocytes 486:509 other myeloid leukocytes 486:509 An inhibitory receptor, FcγRIIb, is expressed on macrophages and other myeloid leukocytes simultaneously with the activating receptor FcγRIIa, thereby setting a threshold for cell activation. 24780636 10 0 from deficiency 1721:1730 arg1 conditions 1756:1765 certain pathological conditions 1735:1765 certain pathological conditions 1735:1765 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 1 1 contain possesses 127:135 arg2 N-glycans 137:145 N-glycans 137:145 N-glycans at Asn432 and Asn630 in humans 137:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 1 1 contain possesses 127:135 arg1 Transferrin 80:90 Transferrin 80:90 Transferrin 80:90 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 1 1 contain possesses 127:135 arg1 protein 113:119 an iron-transport protein 95:119 an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans 95:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 7 2 theme correlation 1156:1166 arg1 spectroscopy 1168:1179 fluorescence correlation spectroscopy 1143:1179 fluorescence correlation spectroscopy 1143:1179 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 2 3 dep glycoforms 191:200 arg1 glycoforms 191:200 Transferrin glycoforms 179:200 Transferrin glycoforms Tf-1 and Tf-2 179:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 3 dep glycoforms 191:200 arg1 Tf-2 211:214 Tf-2 211:214 Tf-2 211:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 3 dep glycoforms 191:200 arg1 Tf-1 202:205 Tf-1 202:205 Tf-1 202:205 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 7 4 theme exclusion 1114:1122 arg1 chromatography 1124:1137 Size exclusion chromatography 1109:1137 Size exclusion chromatography 1109:1137 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 6 5 with glycan 950:955 arg1 GlcNAc 993:998 GlcNAc 993:998 GlcNAc 993:998 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 6 5 with glycan 950:955 arg1 N-acetylglucosamine 972:990 bisecting N-acetylglucosamine 962:990 bisecting N-acetylglucosamine (GlcNAc) 962:999 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 4 6 theme transferrin 648:658 arg1 characterization 607:622 comparative characterization 595:622 comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf) 595:664 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 9 7 theme N-glycan 1521:1528 arg1 structure 1530:1538 the different N-glycan structure 1507:1538 the different N-glycan structure of Tf-1 1507:1546 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 6 8 theme agalacto-complex-type 928:948 arg1 glycan 950:955 agalacto-complex-type glycan 928:955 agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) 928:999 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 8 9 theme exposed 1312:1318 arg1 surface 1332:1338 an exposed hydrophobic surface 1309:1338 an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate 1309:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 5 10 theme spectrometric 672:684 arg1 analyses 686:693 Mass spectrometric analyses 667:693 Mass spectrometric analyses 667:693 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 10 11 theme transferrin 1855:1865 arg1 function 1826:1833 function 1826:1833 function 1826:1833 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 9 12 from surface 1620:1626 arg1 transferrin 1631:1641 transferrin 1631:1641 transferrin 1631:1641 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 1 13 theme iron-transport 98:111 arg1 Transferrin 80:90 Transferrin 80:90 Transferrin 80:90 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 1 13 theme iron-transport 98:111 arg1 protein 113:119 an iron-transport protein 95:119 an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans 95:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 4 14 theme comparative 595:605 arg1 characterization 607:622 comparative characterization 595:622 comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf) 595:664 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 4 15 dep transferrin 535:545 arg1 glycoform 547:555 glycoform 547:555 glycoform 547:555 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 4 15 dep transferrin 535:545 arg1 function 572:579 biological function 561:579 biological function 561:579 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 2 16 theme cerebrospinal 248:260 arg1 fluid 262:266 human cerebrospinal fluid 242:266 human cerebrospinal fluid 242:266 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 9 17 theme apparent 1559:1566 arg1 volume 1578:1583 the apparent hydration volume 1555:1583 the apparent hydration volume 1555:1583 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 2 18 gly glycoforms 191:200 arg1 Transferrin 179:189 Transferrin glycoforms 179:200 Transferrin glycoforms Tf-1 and Tf-2 179:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 1 19 from Asn630 161:166 arg1 N-glycans 137:145 N-glycans 137:145 N-glycans at Asn432 and Asn630 in humans 137:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 5 20 theme disialylated 732:743 arg1 glycans 757:763 disialylated biantennary glycans 732:763 disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf 732:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 1 21 from Asn432 150:155 arg1 N-glycans 137:145 N-glycans 137:145 N-glycans at Asn432 and Asn630 in humans 137:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 1 22 from N-glycans 137:145 arg1 humans 171:176 humans 171:176 humans 171:176 Transferrin is an iron-transport protein which possesses N-glycans at Asn432 and Asn630 in humans. 24780636 5 23 gly disialylated 732:743 arg1 glycans 757:763 disialylated biantennary glycans 732:763 disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf 732:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 3 24 theme hydrocephalus 421:433 arg1 patients 435:442 idiopathic normal pressure hydrocephalus patients 394:442 idiopathic normal pressure hydrocephalus patients 394:442 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 0 25 theme Structural 0:9 arg1 change 11:16 Structural change 0:16 Structural change of N-glycan 0:28 Structural change of N-glycan exposes hydrophobic surface of human transferrin. 24780636 9 26 theme hydrophobic 1608:1618 arg1 surface 1620:1626 hydrophobic surface 1608:1626 hydrophobic surface on transferrin 1608:1641 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 3 27 theme Tf-2/Tf-1 365:373 arg1 ratio 375:379 the Tf-2/Tf-1 ratio 361:379 the Tf-2/Tf-1 ratio 361:379 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 3 27 theme Tf-2/Tf-1 365:373 arg1 useful 451:456 useful 451:456 useful 451:456 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 5 28 mod modified 718:725 arg1 Tf-2 710:713 Tf-2 710:713 Tf-2 710:713 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 5 28 mod modified 718:725 arg3 glycans 757:763 disialylated biantennary glycans 732:763 disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf 732:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 8 29 theme probe 1415:1419 arg1 wavelength 1387:1396 wavelength 1387:1396 wavelength 1387:1396 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 8 29 theme probe 1415:1419 arg1 intensity 1373:1381 fluorescence intensity 1360:1381 fluorescence intensity 1360:1381 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 7 30 theme hydration 1208:1216 arg1 smaller 1245:1251 smaller 1245:1251 smaller 1245:1251 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 7 30 theme hydration 1208:1216 arg1 volume 1218:1223 the hydration volume 1204:1223 the hydration volume of Tf-1 1204:1231 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 6 31 theme other 857:861 arg1 hand 863:866 the other hand 853:866 the other hand 853:866 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 9 32 from transferrin 1631:1641 arg1 patch 1599:1603 a patch 1597:1603 a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2 1597:1701 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 6 33 theme complex/high 1047:1058 arg1 glycans 1073:1079 complex/high mannose-type glycans 1047:1079 complex/high mannose-type glycans 1047:1079 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 3 34 theme normal 405:410 arg1 patients 435:442 idiopathic normal pressure hydrocephalus patients 394:442 idiopathic normal pressure hydrocephalus patients 394:442 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 4 35 theme biological 561:570 arg1 function 572:579 biological function 561:579 biological function 561:579 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 2 36 theme Transferrin 179:189 arg1 bands 304:308 the lower and upper bands 284:308 the lower and upper bands in gel electrophoresis 284:331 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 36 theme Transferrin 179:189 arg1 glycoforms 191:200 Transferrin glycoforms 179:200 Transferrin glycoforms Tf-1 and Tf-2 179:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 36 theme Transferrin 179:189 arg1 Tf-2 211:214 Tf-2 211:214 Tf-2 211:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 36 theme Transferrin 179:189 arg1 Tf-1 202:205 Tf-1 202:205 Tf-1 202:205 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 8 37 theme 1-anilino-8-naphthalene 1422:1444 arg1 sulfonate 1446:1454 1-anilino-8-naphthalene sulfonate 1422:1454 1-anilino-8-naphthalene sulfonate 1422:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 8 37 theme 1-anilino-8-naphthalene 1422:1444 arg1 probe 1415:1419 a hydrophobic probe 1401:1419 a hydrophobic probe 1401:1419 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 10 38 theme carbohydrate 1708:1719 arg1 deficiency 1721:1730 The carbohydrate deficiency 1704:1730 The carbohydrate deficiency in certain pathological conditions 1704:1765 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 5 39 theme N-glycosylation 784:798 arg1 sites 800:804 the two N-glycosylation sites 776:804 the two N-glycosylation sites 776:804 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 5 39 theme N-glycosylation 784:798 arg1 similar 817:823 similar 817:823 similar 817:823 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 0 40 theme human 61:65 arg1 transferrin 67:77 human transferrin 61:77 human transferrin 61:77 Structural change of N-glycan exposes hydrophobic surface of human transferrin. 24780636 5 41 gly N-glycosylation 784:798 arg2 two 780:782 two 780:782 two 780:782 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 5 41 gly N-glycosylation 784:798 arg2 similar 817:823 similar 817:823 similar 817:823 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 5 41 gly N-glycosylation 784:798 arg2 sites 800:804 the two N-glycosylation sites 776:804 the two N-glycosylation sites 776:804 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 10 42 theme pathological 1743:1754 arg1 conditions 1756:1765 certain pathological conditions 1735:1765 certain pathological conditions 1735:1765 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 4 43 theme Tf-1 627:630 arg1 characterization 607:622 comparative characterization 595:622 comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf) 595:664 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 7 44 theme fluorescence 1143:1154 arg1 spectroscopy 1168:1179 fluorescence correlation spectroscopy 1143:1179 fluorescence correlation spectroscopy 1143:1179 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 8 45 contain has 1305:1307 arg2 surface 1332:1338 an exposed hydrophobic surface 1309:1338 an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate 1309:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 8 45 contain has 1305:1307 arg1 Tf-1 1300:1303 Tf-1 1300:1303 Tf-1 1300:1303 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 7 46 theme chromatography 1124:1137 arg1 analysis 1181:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis 1109:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis 1109:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 2 47 theme lower 288:292 arg1 glycoforms 191:200 Transferrin glycoforms 179:200 Transferrin glycoforms Tf-1 and Tf-2 179:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 47 theme lower 288:292 arg1 bands 304:308 the lower and upper bands 284:308 the lower and upper bands in gel electrophoresis 284:331 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 7 48 theme Size 1109:1112 arg1 chromatography 1124:1137 Size exclusion chromatography 1109:1137 Size exclusion chromatography 1109:1137 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 8 49 theme striking 1275:1282 arg1 finding 1284:1290 Our striking finding 1271:1290 Our striking finding 1271:1290 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 2 50 theme upper 298:302 arg1 glycoforms 191:200 Transferrin glycoforms 179:200 Transferrin glycoforms Tf-1 and Tf-2 179:214 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 2 50 theme upper 298:302 arg1 bands 304:308 the lower and upper bands 284:308 the lower and upper bands in gel electrophoresis 284:331 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 6 51 theme bisecting 962:970 arg1 GlcNAc 993:998 GlcNAc 993:998 GlcNAc 993:998 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 6 51 theme bisecting 962:970 arg1 N-acetylglucosamine 972:990 bisecting N-acetylglucosamine 962:990 bisecting N-acetylglucosamine (GlcNAc) 962:999 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 4 52 theme Tf-2 633:636 arg1 characterization 607:622 comparative characterization 595:622 comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf) 595:664 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 2 53 theme human 242:246 arg1 fluid 262:266 human cerebrospinal fluid 242:266 human cerebrospinal fluid 242:266 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 9 54 theme different 1511:1519 arg1 structure 1530:1538 the different N-glycan structure 1507:1538 the different N-glycan structure of Tf-1 1507:1546 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 6 55 with fucose 1010:1015 arg1 GlcNAc 993:998 GlcNAc 993:998 GlcNAc 993:998 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 6 55 with fucose 1010:1015 arg1 N-acetylglucosamine 972:990 bisecting N-acetylglucosamine 962:990 bisecting N-acetylglucosamine (GlcNAc) 962:999 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 4 56 theme serum 642:646 arg1 sTf 661:663 sTf 661:663 sTf 661:663 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 4 56 theme serum 642:646 arg1 transferrin 648:658 serum transferrin 642:658 serum transferrin (sTf) 642:664 In order to gain insight into the relationship between transferrin glycoform and biological function, we performed comparative characterization of Tf-1, Tf-2 and serum transferrin (sTf). 24780636 5 57 theme Mass 667:670 arg1 analyses 686:693 Mass spectrometric analyses 667:693 Mass spectrometric analyses 667:693 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 10 58 dep function 1826:1833 arg1 the 1822:1824 the 1822:1824 the 1822:1824 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 9 59 theme Tf-1 1543:1546 arg1 structure 1530:1538 the different N-glycan structure 1507:1538 the different N-glycan structure of Tf-1 1507:1546 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 8 60 theme hydrophobic 1320:1330 arg1 surface 1332:1338 an exposed hydrophobic surface 1309:1338 an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate 1309:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 0 61 theme transferrin 67:77 arg1 surface 50:56 hydrophobic surface 38:56 hydrophobic surface of human transferrin 38:77 Structural change of N-glycan exposes hydrophobic surface of human transferrin. 24780636 5 62 from both 768:771 arg1 glycans 757:763 disialylated biantennary glycans 732:763 disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf 732:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 9 63 theme hydration 1568:1576 arg1 volume 1578:1583 the apparent hydration volume 1555:1583 the apparent hydration volume 1555:1583 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 9 64 from patch 1599:1603 arg1 transferrin 1631:1641 transferrin 1631:1641 transferrin 1631:1641 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 6 65 theme core 1005:1008 arg1 fucose 1010:1015 core fucose 1005:1015 core fucose 1005:1015 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 2 66 from bands 304:308 arg1 electrophoresis 317:331 gel electrophoresis 313:331 gel electrophoresis 313:331 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 8 67 theme fluorescence 1360:1371 arg1 intensity 1373:1381 fluorescence intensity 1360:1381 fluorescence intensity 1360:1381 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 2 68 theme gel 313:315 arg1 electrophoresis 317:331 gel electrophoresis 313:331 gel electrophoresis 313:331 Transferrin glycoforms Tf-1 and Tf-2, previously identified in human cerebrospinal fluid, are defined as the lower and upper bands in gel electrophoresis, respectively. 24780636 5 69 theme biantennary 745:755 arg1 glycans 757:763 disialylated biantennary glycans 732:763 disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf 732:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 6 70 theme single 1094:1099 arg1 GlcNAc 1101:1106 single GlcNAc 1094:1106 single GlcNAc 1094:1106 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 7 71 theme Tf-1 1228:1231 arg1 smaller 1245:1251 smaller 1245:1251 smaller 1245:1251 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 7 71 theme Tf-1 1228:1231 arg1 volume 1218:1223 the hydration volume 1204:1223 the hydration volume of Tf-1 1204:1231 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 0 72 theme N-glycan 21:28 arg1 change 11:16 Structural change 0:16 Structural change of N-glycan 0:28 Structural change of N-glycan exposes hydrophobic surface of human transferrin. 24780636 9 73 theme surface 1620:1626 arg1 patch 1599:1603 a patch 1597:1603 a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2 1597:1701 These results suggest that the different N-glycan structure of Tf-1 lowers the apparent hydration volume and reveals a patch of hydrophobic surface on transferrin which is otherwise covered with sialoglycan in sTf and Tf-2. 24780636 6 74 theme mannose-type 1060:1071 arg1 glycans 1073:1079 complex/high mannose-type glycans 1047:1079 complex/high mannose-type glycans 1047:1079 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 8 75 theme hydrophobic 1403:1413 arg1 sulfonate 1446:1454 1-anilino-8-naphthalene sulfonate 1422:1454 1-anilino-8-naphthalene sulfonate 1422:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 8 75 theme hydrophobic 1403:1413 arg1 probe 1415:1419 a hydrophobic probe 1401:1419 a hydrophobic probe 1401:1419 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 3 76 theme clinical 463:470 arg1 marker 472:477 a clinical marker 461:477 a clinical marker 461:477 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 0 77 theme hydrophobic 38:48 arg1 surface 50:56 hydrophobic surface 38:56 hydrophobic surface of human transferrin 38:77 Structural change of N-glycan exposes hydrophobic surface of human transferrin. 24780636 8 78 dep intensity 1373:1381 arg1 the 1356:1358 the 1356:1358 the 1356:1358 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 3 79 theme pressure 412:419 arg1 patients 435:442 idiopathic normal pressure hydrocephalus patients 394:442 idiopathic normal pressure hydrocephalus patients 394:442 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 7 80 theme spectroscopy 1168:1179 arg1 analysis 1181:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis 1109:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis 1109:1188 Size exclusion chromatography and fluorescence correlation spectroscopy analysis revealed that the hydration volume of Tf-1 is slightly smaller than that of sTf. 24780636 3 81 theme idiopathic 394:403 arg1 patients 435:442 idiopathic normal pressure hydrocephalus patients 394:442 idiopathic normal pressure hydrocephalus patients 394:442 Importantly, the Tf-2/Tf-1 ratio is raised in idiopathic normal pressure hydrocephalus patients and is useful as a clinical marker. 24780636 8 82 dep surface 1332:1338 arg1 monitored 1343:1351 monitored 1343:1351 monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate 1343:1454 Our striking finding is that Tf-1 has an exposed hydrophobic surface as monitored by the fluorescence intensity and wavelength of a hydrophobic probe, 1-anilino-8-naphthalene sulfonate, whereas Tf-2 does not. 24780636 10 83 theme certain 1735:1741 arg1 conditions 1756:1765 certain pathological conditions 1735:1765 certain pathological conditions 1735:1765 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 5 84 theme sTf 845:847 arg1 N-glycans 832:840 the N-glycans 828:840 the N-glycans of sTf 828:847 Mass spectrometric analyses confirmed that Tf-2 is modified with disialylated biantennary glycans at both of the two N-glycosylation sites, which are similar to the N-glycans of sTf. 24780636 10 85 theme hydrophobic 1783:1793 arg1 surface 1795:1801 hydrophobic surface 1783:1801 hydrophobic surface which may modulate the function and/or stability of transferrin 1783:1865 The carbohydrate deficiency in certain pathological conditions may also expose hydrophobic surface which may modulate the function and/or stability of transferrin. 24780636 6 86 mod modified 1033:1040 arg3 GlcNAc 1101:1106 single GlcNAc 1094:1106 single GlcNAc 1094:1106 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 6 86 mod modified 1033:1040 arg3 glycans 1073:1079 complex/high mannose-type glycans 1047:1079 complex/high mannose-type glycans 1047:1079 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 24780636 6 86 mod modified 1033:1040 arg1 Asn432 1023:1028 Asn432 1023:1028 Asn432 1023:1028 On the other hand, Tf-1 is site-specifically modified: Asn630 has biantennary agalacto-complex-type glycan with bisecting N-acetylglucosamine (GlcNAc) and core fucose while Asn432 is modified with complex/high mannose-type glycans and possibly single GlcNAc. 26249352 5 0 from members 855:861 arg1 part 805:808 part 805:808 part of the catalytic cleft in other GH family 17 members 805:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 0 from members 855:861 arg1 region 780:785 the region 776:785 the region near helix α6 776:799 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 6 1 theme +2 1041:1042 arg1 terminus 1021:1028 the reducing terminus 1008:1028 the reducing terminus of subsite +2 of RmBgt17A 1008:1054 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 10 2 theme antifungal 1770:1779 arg1 screening 1786:1794 antifungal drug screening 1770:1794 antifungal drug screening 1770:1794 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 5 3 from RmBgt17A 877:884 arg1 absent 867:872 absent 867:872 absent 867:872 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 3 from RmBgt17A 877:884 arg1 subdomain 755:763 a conserved subdomain 743:763 a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members 743:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 6 4 theme reducing 1012:1019 arg1 terminus 1021:1028 the reducing terminus 1008:1028 the reducing terminus of subsite +2 of RmBgt17A 1008:1054 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 10 5 theme β-1,3-glucanosyltransferase 1606:1632 arg1 structure 1578:1586 The first crystal structure 1560:1586 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase 1560:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 5 theme β-1,3-glucanosyltransferase 1606:1632 arg1 useful 1641:1646 useful 1641:1646 useful 1641:1646 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 5 6 theme helix 792:796 arg1 α6 798:799 helix α6 792:799 helix α6 792:799 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 10 7 theme family 1596:1601 arg1 β-1,3-glucanosyltransferase 1606:1632 a GH family 17 β-1,3-glucanosyltransferase 1591:1632 a GH family 17 β-1,3-glucanosyltransferase 1591:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 9 8 theme β-1,3-glucan 1537:1548 arg1 acceptor 1550:1557 another β-1,3-glucan acceptor 1529:1557 another β-1,3-glucan acceptor 1529:1557 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 6 9 theme catalytic 1081:1089 arg1 cleft 1091:1095 the catalytic cleft 1077:1095 the catalytic cleft 1077:1095 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 4 10 theme RmBgt17A 596:603 arg1 structure 583:591 The overall structure 571:591 The overall structure of RmBgt17A 571:603 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 3 11 theme Å 554:554 arg1 resolutions 519:529 resolutions 519:529 resolutions of 1.30, 2.30 and 2.27 Å 519:554 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 5 12 from located 765:771 arg1 part 805:808 part 805:808 part of the catalytic cleft in other GH family 17 members 805:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 12 from located 765:771 arg1 region 780:785 the region 776:785 the region near helix α6 776:799 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 2 13 theme branched 211:218 arg1 glucans 220:226 branched glucans 211:226 branched glucans 211:226 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 5 14 theme catalytic 817:825 arg1 cleft 827:831 the catalytic cleft 813:831 the catalytic cleft in other GH family 17 members 813:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 10 15 theme mechanism 1676:1684 arg1 studies 1651:1657 studies 1651:1657 studies of the catalytic mechanism of GH family 17 proteins 1651:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 7 16 theme family 1197:1202 arg1 enzymes 1207:1213 other reported GH family 17 enzymes 1179:1213 other reported GH family 17 enzymes 1179:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 10 17 theme catalytic 1666:1674 arg1 mechanism 1676:1684 the catalytic mechanism 1662:1684 the catalytic mechanism of GH family 17 proteins 1662:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 9 18 theme clear 1342:1346 arg1 explanation 1348:1358 a clear explanation 1340:1358 a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor 1340:1557 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 7 19 theme reported 1185:1192 arg1 enzymes 1207:1213 other reported GH family 17 enzymes 1179:1213 other reported GH family 17 enzymes 1179:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 5 20 theme family 692:697 arg1 members 702:708 other GH family 17 members 683:708 other GH family 17 members 683:708 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 4 21 theme TIM-barrel 635:644 arg1 fold 646:649 the characteristic (β/α)8 TIM-barrel fold 609:649 the characteristic (β/α)8 TIM-barrel fold 609:649 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 5 22 from part 805:808 arg1 located 765:771 located 765:771 located 765:771 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 22 from part 805:808 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 9 23 theme mode 1377:1380 arg1 explanation 1348:1358 a clear explanation 1340:1358 a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor 1340:1557 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 3 24 theme glycoside 335:343 arg1 family 360:365 a novel glycoside hydrolase (GH) family 17 327:368 a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 327:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 5 25 theme RmBgt17A 670:677 arg1 members 702:708 other GH family 17 members 683:708 other GH family 17 members 683:708 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 25 theme RmBgt17A 670:677 arg1 structures 656:665 The structures 652:665 The structures of RmBgt17A 652:677 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 9 26 theme RmBgt17A 1385:1392 arg1 mode 1377:1380 the catalytic mode 1363:1380 the catalytic mode 1363:1380 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 10 27 from useful 1641:1646 arg1 studies 1651:1657 studies 1651:1657 studies of the catalytic mechanism of GH family 17 proteins 1651:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 5 28 theme other 683:687 arg1 members 702:708 other GH family 17 members 683:708 other GH family 17 members 683:708 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 29 from cleft 827:831 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 10 30 from studies 1651:1657 arg1 structure 1578:1586 The first crystal structure 1560:1586 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase 1560:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 30 from studies 1651:1657 arg1 useful 1641:1646 useful 1641:1646 useful 1641:1646 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 7 31 theme other 1179:1183 arg1 enzymes 1207:1213 other reported GH family 17 enzymes 1179:1213 other reported GH family 17 enzymes 1179:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 5 32 theme GH 842:843 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 2 33 theme important 176:184 arg1 role 186:189 an important role 173:189 an important role 173:189 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 6 34 theme RmBgt17A 1047:1054 arg1 +2 1041:1042 subsite +2 1033:1042 subsite +2 of RmBgt17A 1033:1054 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 8 35 theme complex 1220:1226 arg1 structures 1228:1237 The complex structures 1216:1237 The complex structures 1216:1237 The complex structures also illustrated that RmBgt17A can only provide subsites -3 to +2. 26249352 0 36 theme crystal 10:16 arg1 structure 18:26 The first crystal structure 0:26 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 0:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 0 37 dep family 53:58 arg1 β-1,3-glucanosyltransferase 63:89 β-1,3-glucanosyltransferase 63:89 a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 31:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 7 38 theme shorter 1157:1163 arg1 cleft 1151:1155 its catalytic cleft 1137:1155 its catalytic cleft shorter than those of other reported GH family 17 enzymes 1137:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 10 39 theme family 1692:1697 arg1 proteins 1702:1709 GH family 17 proteins 1689:1709 GH family 17 proteins 1689:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 7 40 theme catalytic 1141:1149 arg1 cleft 1151:1155 its catalytic cleft 1137:1155 its catalytic cleft shorter than those of other reported GH family 17 enzymes 1137:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 0 41 theme family 53:58 arg1 structure 18:26 The first crystal structure 0:26 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 0:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 3 42 theme mutant 469:474 arg1 structures 313:322 The crystal structures 301:322 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 301:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 42 theme mutant 469:474 arg1 complexes 440:448 the complexes 436:448 the complexes of its active-site mutant (E189A) with two substrates 436:502 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 0 43 theme glycoside 33:41 arg1 family 53:58 a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 31:89 a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 31:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 9 44 theme linear 1455:1460 arg1 β-1,3-glucan 1462:1473 linear β-1,3-glucan 1455:1473 linear β-1,3-glucan 1455:1473 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 10 45 theme proteins 1702:1709 arg1 mechanism 1676:1684 the catalytic mechanism 1662:1684 the catalytic mechanism of GH family 17 proteins 1662:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 46 theme crystal 1570:1576 arg1 structure 1578:1586 The first crystal structure 1560:1586 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase 1560:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 46 theme crystal 1570:1576 arg1 useful 1641:1646 useful 1641:1646 useful 1641:1646 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 7 47 theme distinct 1103:1110 arg1 region 1112:1117 This distinct region 1098:1117 This distinct region of RmBgt17A 1098:1129 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 9 48 theme remaining 1483:1491 arg1 glucan 1493:1498 the remaining glucan 1479:1498 the remaining glucan 1479:1498 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 10 49 theme GH 1593:1594 arg1 β-1,3-glucanosyltransferase 1606:1632 a GH family 17 β-1,3-glucanosyltransferase 1591:1632 a GH family 17 β-1,3-glucanosyltransferase 1591:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 6 50 theme subsite 1033:1039 arg1 +2 1041:1042 subsite +2 1033:1042 subsite +2 of RmBgt17A 1033:1054 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 9 51 attach released 1421:1428 arg2 laminaribiose 1404:1416 laminaribiose 1404:1416 laminaribiose 1404:1416 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 9 51 attach released 1421:1428 arg1 end 1448:1450 the reducing end 1435:1450 the reducing end of linear β-1,3-glucan 1435:1473 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 10 52 theme enzymatic 1745:1753 arg1 engineering 1755:1765 further enzymatic engineering 1737:1765 further enzymatic engineering 1737:1765 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 3 53 with mutant 469:474 arg1 substrates 493:502 two substrates 489:502 two substrates 489:502 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 4 54 theme characteristic 613:626 arg1 fold 646:649 the characteristic (β/α)8 TIM-barrel fold 609:649 the characteristic (β/α)8 TIM-barrel fold 609:649 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 5 55 theme conserved 745:753 arg1 absent 867:872 absent 867:872 absent 867:872 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 55 theme conserved 745:753 arg1 subdomain 755:763 a conserved subdomain 743:763 a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members 743:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 4 56 theme overall 575:581 arg1 structure 583:591 The overall structure 571:591 The overall structure of RmBgt17A 571:603 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 10 57 theme drug 1781:1784 arg1 screening 1786:1794 antifungal drug screening 1770:1794 antifungal drug screening 1770:1794 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 5 58 theme located 765:771 arg1 absent 867:872 absent 867:872 absent 867:872 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 58 theme located 765:771 arg1 subdomain 755:763 a conserved subdomain 743:763 a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members 743:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 9 59 theme acceptor 1550:1557 arg1 end 1522:1524 the end 1518:1524 the end of another β-1,3-glucan acceptor 1518:1557 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 2 60 theme glucans 220:226 arg1 formation 198:206 the formation 194:206 the formation of branched glucans 194:226 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 2 61 theme cell-wall 243:251 arg1 assembly 253:260 cell-wall assembly 243:260 cell-wall assembly 243:260 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 5 62 theme cleft 827:831 arg1 part 805:808 part 805:808 part of the catalytic cleft in other GH family 17 members 805:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 62 theme cleft 827:831 arg1 region 780:785 the region 776:785 the region near helix α6 776:799 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 2 63 from assembly 253:260 arg1 yeasts 293:298 yeasts 293:298 yeasts 293:298 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 2 63 from assembly 253:260 arg1 fungi 283:287 fungi 283:287 fungi 283:287 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 5 64 theme GH 689:690 arg1 members 702:708 other GH family 17 members 683:708 other GH family 17 members 683:708 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 2 65 from rearrangement 266:278 arg1 yeasts 293:298 yeasts 293:298 yeasts 293:298 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 2 65 from rearrangement 266:278 arg1 fungi 283:287 fungi 283:287 fungi 283:287 -) plays an important role in the formation of branched glucans, as well as in cell-wall assembly and rearrangement in fungi and yeasts. 26249352 6 66 theme amino-acid 901:910 arg1 His172 986:991 His172 986:991 His172 986:991 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 66 theme amino-acid 901:910 arg1 Glu158 975:980 Glu158 975:980 Glu158 975:980 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 66 theme amino-acid 901:910 arg1 residues 912:919 four amino-acid residues 896:919 four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) 896:992 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 66 theme amino-acid 901:910 arg1 Tyr136 967:972 Tyr136 967:972 Tyr136 967:972 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 66 theme amino-acid 901:910 arg1 Tyr135 959:964 Tyr135 959:964 Tyr135 959:964 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 7 67 theme GH 1194:1195 arg1 enzymes 1207:1213 other reported GH family 17 enzymes 1179:1213 other reported GH family 17 enzymes 1179:1213 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 4 68 theme β/α 629:631 arg1 fold 646:649 the characteristic (β/α)8 TIM-barrel fold 609:649 the characteristic (β/α)8 TIM-barrel fold 609:649 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 5 69 from region 780:785 arg1 located 765:771 located 765:771 located 765:771 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 5 69 from region 780:785 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 9 70 theme catalytic 1367:1375 arg1 mode 1377:1380 the catalytic mode 1363:1380 the catalytic mode 1363:1380 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 3 71 theme crystal 305:311 arg1 structures 313:322 The crystal structures 301:322 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 301:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 0 72 theme unique 102:107 arg1 cleft 119:123 a unique catalytic cleft 100:123 a unique catalytic cleft 100:123 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 3 73 theme hydrolase 345:353 arg1 family 360:365 a novel glycoside hydrolase (GH) family 17 327:368 a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 327:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 74 theme novel 329:333 arg1 family 360:365 a novel glycoside hydrolase (GH) family 17 327:368 a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 327:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 6 75 dep exposed 921:927 arg1 His172 986:991 His172 986:991 His172 986:991 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 75 dep exposed 921:927 arg1 Glu158 975:980 Glu158 975:980 Glu158 975:980 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 75 dep exposed 921:927 arg1 residues 912:919 four amino-acid residues 896:919 four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) 896:992 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 75 dep exposed 921:927 arg1 Tyr136 967:972 Tyr136 967:972 Tyr136 967:972 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 75 dep exposed 921:927 arg1 Tyr135 959:964 Tyr135 959:964 Tyr135 959:964 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 1 76 dep β-1,3-Glucanosyltransferase 126:152 arg1 EC 155:156 EC 2.4.1 155:162 EC 2.4.1 155:162 β-1,3-Glucanosyltransferase (EC 2.4.1. 26249352 0 77 theme first 4:8 arg1 structure 18:26 The first crystal structure 0:26 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 0:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 5 78 theme other 836:840 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 6 79 theme enzyme 951:956 arg1 surface 936:942 the surface 932:942 the surface of the enzyme 932:956 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 3 80 theme family 360:365 arg1 structures 313:322 The crystal structures 301:322 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 301:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 80 theme family 360:365 arg1 complexes 440:448 the complexes 436:448 the complexes of its active-site mutant (E189A) with two substrates 436:502 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 5 81 theme family 845:850 arg1 members 855:861 other GH family 17 members 836:861 other GH family 17 members 836:861 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26249352 3 82 theme GH 356:357 arg1 family 360:365 a novel glycoside hydrolase (GH) family 17 327:368 a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 327:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 9 83 theme structural 1311:1320 arg1 evidence 1322:1329 This structural evidence 1306:1329 This structural evidence 1306:1329 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 3 84 theme Rhizomucor 403:412 arg1 RmBgt17A 422:429 RmBgt17A 422:429 RmBgt17A 422:429 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 84 theme Rhizomucor 403:412 arg1 miehei 414:419 Rhizomucor miehei 403:419 Rhizomucor miehei (RmBgt17A) 403:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 10 85 theme GH 1689:1690 arg1 proteins 1702:1709 GH family 17 proteins 1689:1709 GH family 17 proteins 1689:1709 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 0 86 theme catalytic 109:117 arg1 cleft 119:123 a unique catalytic cleft 100:123 a unique catalytic cleft 100:123 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 0 87 theme hydrolase 43:51 arg1 family 53:58 a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 31:89 a glycoside hydrolase family 17 β-1,3-glucanosyltransferase 31:89 The first crystal structure of a glycoside hydrolase family 17 β-1,3-glucanosyltransferase displays a unique catalytic cleft. 26249352 9 88 theme reducing 1439:1446 arg1 end 1448:1450 the reducing end 1435:1450 the reducing end of linear β-1,3-glucan 1435:1473 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 7 89 theme RmBgt17A 1122:1129 arg1 region 1112:1117 This distinct region 1098:1117 This distinct region of RmBgt17A 1098:1129 This distinct region of RmBgt17A makes its catalytic cleft shorter than those of other reported GH family 17 enzymes. 26249352 3 90 theme β-1,3-glucanosyltransferase 370:396 arg1 family 360:365 a novel glycoside hydrolase (GH) family 17 327:368 a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) 327:430 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 8 91 dep subsites 1287:1294 arg1 to 1299:1300 to 1299:1300 to 1299:1300 The complex structures also illustrated that RmBgt17A can only provide subsites -3 to +2. 26249352 4 92 contain had 605:607 arg1 structure 583:591 The overall structure 571:591 The overall structure of RmBgt17A 571:603 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 4 92 contain had 605:607 arg2 fold 646:649 the characteristic (β/α)8 TIM-barrel fold 609:649 the characteristic (β/α)8 TIM-barrel fold 609:649 The overall structure of RmBgt17A had the characteristic (β/α)8 TIM-barrel fold. 26249352 3 93 from miehei 414:419 arg1 β-1,3-glucanosyltransferase 370:396 β-1,3-glucanosyltransferase 370:396 β-1,3-glucanosyltransferase 370:396 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 94 theme active-site 457:467 arg1 mutant 469:474 its active-site mutant 453:474 its active-site mutant (E189A) with two substrates 453:502 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 3 94 theme active-site 457:467 arg1 E189A 477:481 E189A 477:481 E189A 477:481 The crystal structures of a novel glycoside hydrolase (GH) family 17 β-1,3-glucanosyltransferase from Rhizomucor miehei (RmBgt17A) and the complexes of its active-site mutant (E189A) with two substrates were solved at resolutions of 1.30, 2.30 and 2.27 Å, respectively. 26249352 9 95 theme β-1,3-glucan 1462:1473 arg1 end 1448:1450 the reducing end 1435:1450 the reducing end of linear β-1,3-glucan 1435:1473 This structural evidence provides a clear explanation of the catalytic mode of RmBgt17A, in which laminaribiose is released from the reducing end of linear β-1,3-glucan and the remaining glucan is transferred to the end of another β-1,3-glucan acceptor. 26249352 6 96 located found 999:1003 arg2 residues 912:919 four amino-acid residues 896:919 four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) 896:992 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 96 located found 999:1003 arg2 Glu158 975:980 Glu158 975:980 Glu158 975:980 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 96 located found 999:1003 arg1 terminus 1021:1028 the reducing terminus 1008:1028 the reducing terminus of subsite +2 of RmBgt17A 1008:1054 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 96 located found 999:1003 arg2 Tyr135 959:964 Tyr135 959:964 Tyr135 959:964 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 96 located found 999:1003 arg2 His172 986:991 His172 986:991 His172 986:991 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 6 96 located found 999:1003 arg2 Tyr136 967:972 Tyr136 967:972 Tyr136 967:972 Instead, four amino-acid residues exposed to the surface of the enzyme (Tyr135, Tyr136, Glu158 and His172) were found in the reducing terminus of subsite +2 of RmBgt17A, hindering access to the catalytic cleft. 26249352 10 97 theme further 1737:1743 arg1 engineering 1755:1765 further enzymatic engineering 1737:1765 further enzymatic engineering 1737:1765 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 98 theme first 1564:1568 arg1 structure 1578:1586 The first crystal structure 1560:1586 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase 1560:1632 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 10 98 theme first 1564:1568 arg1 useful 1641:1646 useful 1641:1646 useful 1641:1646 The first crystal structure of a GH family 17 β-1,3-glucanosyltransferase may be useful in studies of the catalytic mechanism of GH family 17 proteins, and provides a basis for further enzymatic engineering or antifungal drug screening. 26249352 5 99 from absent 867:872 arg1 RmBgt17A 877:884 RmBgt17A 877:884 RmBgt17A 877:884 The structures of RmBgt17A and other GH family 17 members were compared: it was found that a conserved subdomain located in the region near helix α6 and part of the catalytic cleft in other GH family 17 members was absent in RmBgt17A. 26773038 2 0 theme disulfide 393:401 arg1 bond 403:406 a vicinal cysteine disulfide bond 374:406 a vicinal cysteine disulfide bond 374:406 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 8 1 theme aspartic 1154:1161 arg1 acid 1163:1166 aspartic acid 1154:1166 aspartic acid 1154:1166 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 6 2 theme N1574 954:958 arg1 site 971:974 the N1574 attachment site 950:974 the N1574 attachment site 950:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 1 3 theme factor 136:141 arg1 domain 152:157 the von Willebrand factor (VWF) A2 domain 117:157 the von Willebrand factor (VWF) A2 domain 117:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 6 4 theme sugar 924:928 arg1 component 877:885 The essential component 863:885 The essential component of the glycan structure 863:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 4 theme sugar 924:928 arg1 GlcNAc 939:944 GlcNAc 939:944 GlcNAc 939:944 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 4 theme sugar 924:928 arg1 residue 930:936 the first sugar residue 914:936 the first sugar residue (GlcNAc) at the N1574 attachment site 914:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 2 5 theme cysteine 384:391 arg1 bond 403:406 a vicinal cysteine disulfide bond 374:406 a vicinal cysteine disulfide bond 374:406 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 6 6 theme first 918:922 arg1 component 877:885 The essential component 863:885 The essential component of the glycan structure 863:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 6 theme first 918:922 arg1 GlcNAc 939:944 GlcNAc 939:944 GlcNAc 939:944 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 6 theme first 918:922 arg1 residue 930:936 the first sugar residue 914:936 the first sugar residue (GlcNAc) at the N1574 attachment site 914:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 3 7 with interaction 488:498 arg1 ADAMTS13 505:512 ADAMTS13 505:512 ADAMTS13 505:512 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 9 8 theme cysteine 1406:1413 arg1 bond 1425:1428 vicinal cysteine disulfide bond 1398:1428 vicinal cysteine disulfide bond 1398:1428 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 6 9 from site 971:974 arg1 component 877:885 The essential component 863:885 The essential component of the glycan structure 863:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 9 from site 971:974 arg1 GlcNAc 939:944 GlcNAc 939:944 GlcNAc 939:944 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 9 from site 971:974 arg1 residue 930:936 the first sugar residue 914:936 the first sugar residue (GlcNAc) at the N1574 attachment site 914:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 4 10 link N-linked 600:607 arg1 glycans 609:615 the N-linked glycans 596:615 the N-linked glycans of the VWF A2 domain 596:636 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 9 11 theme VWF 1338:1340 arg1 domain 1345:1350 the VWF A2 domain 1334:1350 the VWF A2 domain 1334:1350 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 6 12 theme glycan 894:899 arg1 structure 901:909 the glycan structure 890:909 the glycan structure 890:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 9 13 theme binding 1381:1387 arg1 site 1389:1392 the Ca(2+)binding site 1371:1392 the Ca(2+)binding site 1371:1392 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 3 14 from N1574 426:430 arg1 Glycosylation 409:421 Glycosylation 409:421 Glycosylation at N1574 409:430 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 1 15 theme Shear 52:56 arg1 forces 58:63 Shear forces 52:63 Shear forces in the blood 52:76 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 4 16 theme domain 631:636 arg1 glycans 609:615 the N-linked glycans 596:615 the N-linked glycans of the VWF A2 domain 596:636 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 4 17 theme A2 628:629 arg1 domain 631:636 the VWF A2 domain 620:636 the VWF A2 domain 620:636 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 0 18 link N-linked 0:7 arg1 stabilization 16:28 N-linked glycan stabilization 0:28 N-linked glycan stabilization of the VWF A2 domain 0:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 9 19 theme A2 1342:1343 arg1 domain 1345:1350 the VWF A2 domain 1334:1350 the VWF A2 domain 1334:1350 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 5 20 theme scanning 769:776 arg1 fluorimetry 778:788 differential scanning fluorimetry 756:788 differential scanning fluorimetry 756:788 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 7 21 theme intradomain 1052:1062 arg1 interactions 1064:1075 stabilizing intradomain interactions 1040:1075 stabilizing intradomain interactions 1040:1075 From its crystal structures, N1574-GlcNAc is predicted to form stabilizing intradomain interactions with Y1544 and nearby residues. 26773038 0 22 theme VWF 37:39 arg1 domain 44:49 the VWF A2 domain 33:49 the VWF A2 domain 33:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 9 23 theme vicinal 1398:1404 arg1 bond 1425:1428 vicinal cysteine disulfide bond 1398:1428 vicinal cysteine disulfide bond 1398:1428 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 3 24 gly Glycosylation 409:421 arg2 N1574 426:430 N1574 426:430 N1574 426:430 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 2 25 theme conformational 310:323 arg1 transition 325:334 The conformational transition 306:334 The conformational transition 306:334 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 2 26 theme binding 357:363 arg1 site 365:368 a Ca(2+)binding site 349:368 a Ca(2+)binding site 349:368 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 7 27 theme crystal 986:992 arg1 structures 994:1003 its crystal structures 982:1003 its crystal structures 982:1003 From its crystal structures, N1574-GlcNAc is predicted to form stabilizing intradomain interactions with Y1544 and nearby residues. 26773038 3 28 theme A2 478:479 arg1 interaction 488:498 VWF A2 domain interaction 474:498 VWF A2 domain interaction with ADAMTS13 474:512 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 7 29 theme nearby 1092:1097 arg1 residues 1099:1106 nearby residues 1092:1106 nearby residues 1092:1106 From its crystal structures, N1574-GlcNAc is predicted to form stabilizing intradomain interactions with Y1544 and nearby residues. 26773038 1 30 theme A2 149:150 arg1 domain 152:157 the von Willebrand factor (VWF) A2 domain 117:157 the von Willebrand factor (VWF) A2 domain 117:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 0 31 theme N-linked 0:7 arg1 stabilization 16:28 N-linked glycan stabilization 0:28 N-linked glycan stabilization of the VWF A2 domain 0:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 6 32 theme structure 901:909 arg1 component 877:885 The essential component 863:885 The essential component of the glycan structure 863:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 32 theme structure 901:909 arg1 residue 930:936 the first sugar residue 914:936 the first sugar residue (GlcNAc) at the N1574 attachment site 914:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 9 33 theme ADAMTS13 1455:1462 arg1 proteolysis 1464:1474 ADAMTS13 proteolysis 1455:1474 ADAMTS13 proteolysis 1455:1474 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 3 34 theme domain 481:486 arg1 interaction 488:498 VWF A2 domain interaction 474:498 VWF A2 domain interaction with ADAMTS13 474:512 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 1 35 theme cryptic 218:224 arg1 Y1605-M1606 241:251 Y1605-M1606 241:251 Y1605-M1606 241:251 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 35 theme cryptic 218:224 arg1 bond 235:238 the cryptic scissile bond 214:238 the cryptic scissile bond (Y1605-M1606) 214:252 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 36 from transition 103:112 arg1 domain 152:157 the von Willebrand factor (VWF) A2 domain 117:157 the von Willebrand factor (VWF) A2 domain 117:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 8 37 theme Y1544 1145:1149 arg1 Substitution 1109:1120 Substitution 1109:1120 Substitution of the surface-exposed Y1544 to aspartic acid 1109:1166 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 2 38 theme Ca 351:352 arg1 site 365:368 a Ca(2+)binding site 349:368 a Ca(2+)binding site 349:368 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 8 39 theme VWF 1285:1287 arg1 domain 1292:1297 the VWF A2 domain 1281:1297 the VWF A2 domain 1281:1297 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 1 40 theme scissile 226:233 arg1 Y1605-M1606 241:251 Y1605-M1606 241:251 Y1605-M1606 241:251 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 40 theme scissile 226:233 arg1 bond 235:238 the cryptic scissile bond 214:238 the cryptic scissile bond (Y1605-M1606) 214:252 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 3 41 theme VWF 474:476 arg1 interaction 488:498 VWF A2 domain interaction 474:498 VWF A2 domain interaction with ADAMTS13 474:512 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 5 42 theme N-linked 799:806 arg1 glycans 808:814 the N-linked glycans 795:814 the N-linked glycans 795:814 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 2 43 theme vicinal 376:382 arg1 bond 403:406 a vicinal cysteine disulfide bond 374:406 a vicinal cysteine disulfide bond 374:406 The conformational transition depends upon a Ca(2+)binding site and a vicinal cysteine disulfide bond. 26773038 5 44 theme VWF 848:850 arg1 domain 855:860 the VWF A2 domain 844:860 the VWF A2 domain 844:860 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 5 45 link N-linked 799:806 arg1 glycans 808:814 the N-linked glycans 795:814 the N-linked glycans 795:814 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 4 46 theme scissile 730:737 arg1 bond 739:742 the scissile bond 726:742 the scissile bond 726:742 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 9 47 theme disulfide 1415:1423 arg1 bond 1425:1428 vicinal cysteine disulfide bond 1398:1428 vicinal cysteine disulfide bond 1398:1428 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 8 48 theme surface-exposed 1129:1143 arg1 Y1544 1145:1149 the surface-exposed Y1544 1125:1149 the surface-exposed Y1544 1125:1149 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 3 49 theme bulky 546:550 arg1 structure 565:573 the bulky carbohydrate structure 542:573 the bulky carbohydrate structure 542:573 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 5 50 theme differential 756:767 arg1 fluorimetry 778:788 differential scanning fluorimetry 756:788 differential scanning fluorimetry 756:788 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 0 51 theme domain 44:49 arg1 stabilization 16:28 N-linked glycan stabilization 0:28 N-linked glycan stabilization of the VWF A2 domain 0:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 6 52 theme essential 867:875 arg1 component 877:885 The essential component 863:885 The essential component of the glycan structure 863:909 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 6 52 theme essential 867:875 arg1 residue 930:936 the first sugar residue 914:936 the first sugar residue (GlcNAc) at the N1574 attachment site 914:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 8 53 theme A2 1289:1290 arg1 domain 1292:1297 the VWF A2 domain 1281:1297 the VWF A2 domain 1281:1297 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 3 54 theme carbohydrate 552:563 arg1 structure 565:573 the bulky carbohydrate structure 542:573 the bulky carbohydrate structure 542:573 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 9 55 theme domain 1345:1350 arg1 stabilization 1317:1329 Glycan stabilization 1310:1329 Glycan stabilization of the VWF A2 domain 1310:1350 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 1 56 theme conformational 88:101 arg1 transition 103:112 a conformational transition 86:112 a conformational transition in the von Willebrand factor (VWF) A2 domain 86:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 0 57 theme A2 41:42 arg1 domain 44:49 the VWF A2 domain 33:49 the VWF A2 domain 33:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 9 58 theme Ca 1375:1376 arg1 site 1389:1392 the Ca(2+)binding site 1371:1392 the Ca(2+)binding site 1371:1392 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 4 59 theme VWF 624:626 arg1 domain 631:636 the VWF A2 domain 620:636 the VWF A2 domain 620:636 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 4 60 theme binding 708:714 arg1 sites 716:720 the ADAMTS13 binding sites 695:720 the ADAMTS13 binding sites 695:720 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 4 61 theme N-linked 600:607 arg1 glycans 609:615 the N-linked glycans 596:615 the N-linked glycans of the VWF A2 domain 596:636 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 5 62 theme A2 852:853 arg1 domain 855:860 the VWF A2 domain 844:860 the VWF A2 domain 844:860 We show by differential scanning fluorimetry that the N-linked glycans thermodynamically stabilize the VWF A2 domain. 26773038 1 63 theme Willebrand 125:134 arg1 VWF 144:146 VWF 144:146 VWF 144:146 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 63 theme Willebrand 125:134 arg1 factor 136:141 the von Willebrand factor 117:141 the von Willebrand factor (VWF) A2 domain 117:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 8 64 theme ADAMTS13 1252:1259 arg1 proteolysis 1261:1271 ADAMTS13 proteolysis 1252:1271 ADAMTS13 proteolysis 1252:1271 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 8 65 theme glycosylation 1218:1230 arg1 absence 1207:1213 the absence 1203:1213 the absence of glycosylation 1203:1230 Substitution of the surface-exposed Y1544 to aspartic acid is able to stabilize the domain in the absence of glycosylation and protect against ADAMTS13 proteolysis in both the VWF A2 domain and FLVWF. 26773038 3 66 theme steric 522:527 arg1 hindrance 529:537 steric hindrance 522:537 steric hindrance 522:537 Glycosylation at N1574 has previously been suggested to modulate VWF A2 domain interaction with ADAMTS13 through steric hindrance by the bulky carbohydrate structure. 26773038 6 67 theme attachment 960:969 arg1 site 971:974 the N1574 attachment site 950:974 the N1574 attachment site 950:974 The essential component of the glycan structure is the first sugar residue (GlcNAc) at the N1574 attachment site. 26773038 1 68 theme von 121:123 arg1 VWF 144:146 VWF 144:146 VWF 144:146 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 68 theme von 121:123 arg1 factor 136:141 the von Willebrand factor 117:141 the von Willebrand factor (VWF) A2 domain 117:157 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 9 69 theme Glycan 1310:1315 arg1 stabilization 1317:1329 Glycan stabilization 1310:1329 Glycan stabilization of the VWF A2 domain 1310:1350 Glycan stabilization of the VWF A2 domain acts together with the Ca(2+)binding site and vicinal cysteine disulfide bond to control unfolding and ADAMTS13 proteolysis. 26773038 7 70 theme stabilizing 1040:1050 arg1 interactions 1064:1075 stabilizing intradomain interactions 1040:1075 stabilizing intradomain interactions 1040:1075 From its crystal structures, N1574-GlcNAc is predicted to form stabilizing intradomain interactions with Y1544 and nearby residues. 26773038 4 71 theme sites 716:720 arg1 exposure 683:690 the exposure 679:690 the exposure of the ADAMTS13 binding sites 679:720 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 4 71 theme sites 716:720 arg1 bond 739:742 the scissile bond 726:742 the scissile bond 726:742 We investigated how the N-linked glycans of the VWF A2 domain affect thermostability and regulate both the exposure of the ADAMTS13 binding sites and the scissile bond. 26773038 0 72 theme glycan 9:14 arg1 stabilization 16:28 N-linked glycan stabilization 0:28 N-linked glycan stabilization of the VWF A2 domain 0:49 N-linked glycan stabilization of the VWF A2 domain. 26773038 1 73 from forces 58:63 arg1 blood 72:76 the blood 68:76 the blood 68:76 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 26773038 1 74 theme unfolded 189:196 arg1 state 198:202 an unfolded state 186:202 an unfolded state 186:202 Shear forces in the blood trigger a conformational transition in the von Willebrand factor (VWF) A2 domain, from its native folded to an unfolded state, in which the cryptic scissile bond (Y1605-M1606) is exposed and can then be proteolysed by ADAMTS13. 28186137 3 0 theme disability 326:335 arg1 degeneration 268:279 Axonal degeneration 261:279 Axonal degeneration 261:279 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 3 0 theme disability 326:335 arg1 cause 294:298 the major cause 284:298 the major cause of permanent neurological disability in primary myelin diseases 284:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 5 1 theme glycans 529:535 arg1 roles 520:524 the roles 516:524 the roles of glycans on myelin glycoproteins 516:559 However, the roles of glycans on myelin glycoproteins remain poorly understood. 28186137 1 2 theme myelin 157:162 arg1 membrane 164:171 a multilayered myelin membrane 142:171 a multilayered myelin membrane in vertebrates 142:186 Highly specialized glial cells wrap axons with a multilayered myelin membrane in vertebrates. 28186137 11 3 dep PNS 1417:1419 arg1 myelination 1421:1431 myelination 1421:1431 myelination 1421:1431 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 11 3 dep PNS 1417:1419 arg1 survival 1455:1462 myelinated axonal survival 1437:1462 myelinated axonal survival 1437:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 11 3 dep PNS 1417:1419 arg1 PNS 1417:1419 PNS myelination and myelinated axonal survival 1417:1462 PNS myelination and myelinated axonal survival 1417:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 7 4 theme abundant 728:735 arg1 N-glycans 746:754 highly abundant sulfated N-glycans 721:754 highly abundant sulfated N-glycans 721:754 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 0 5 theme nervous 79:85 arg1 system 87:92 the peripheral nervous system 64:92 the peripheral nervous system 64:92 GlcNAc6ST-1 regulates sulfation of N-glycans and myelination in the peripheral nervous system. 28186137 9 6 from glycoprotein 1006:1017 arg1 PNS 1022:1024 PNS myelin and mutations 1022:1045 PNS 1022:1024 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 6 from glycoprotein 1006:1017 arg1 mutations 1037:1045 mutations 1037:1045 mutations 1037:1045 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 4 7 theme myelin 434:439 arg1 glycoprotein 441:452 one myelin glycoprotein 430:452 one myelin glycoprotein 430:452 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 9 8 theme glycosylation 1063:1075 arg1 site 1077:1080 the glycosylation site 1059:1080 the glycosylation site 1059:1080 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 7 9 theme sulfated 737:744 arg1 N-glycans 746:754 highly abundant sulfated N-glycans 721:754 highly abundant sulfated N-glycans 721:754 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 9 10 gly glycoprotein 1006:1017 arg1 glycoprotein 1006:1017 the most abundant glycoprotein 988:1017 the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy 988:1117 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 10 gly glycoprotein 1006:1017 arg1 protein 979:985 P0 protein 976:985 P0 protein 976:985 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 1 11 from membrane 164:171 arg1 vertebrates 176:186 vertebrates 176:186 vertebrates 176:186 Highly specialized glial cells wrap axons with a multilayered myelin membrane in vertebrates. 28186137 6 12 theme sulfated 608:615 arg1 N-glycans 617:625 sulfated N-glycans 608:625 sulfated N-glycans 608:625 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 10 13 from deficient 1169:1177 arg1 GlcNAc6ST-1 1226:1236 GlcNAc6ST-1 1226:1236 GlcNAc6ST-1 1226:1236 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 10 13 from deficient 1169:1177 arg1 N-acetylglucosamine-6-O-sulfotransferase-1 1182:1223 N-acetylglucosamine-6-O-sulfotransferase-1 1182:1223 N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) 1182:1237 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 11 14 gly glycoproteins 1513:1525 arg1 glycoproteins 1513:1525 glycoproteins 1513:1525 glycoproteins 1513:1525 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 9 15 theme Charcot-Marie-Tooth 1088:1106 arg1 neuropathy 1108:1117 Charcot-Marie-Tooth neuropathy 1088:1117 Charcot-Marie-Tooth neuropathy 1088:1117 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 16 theme P0 976:977 arg1 glycoprotein 1006:1017 the most abundant glycoprotein 988:1017 the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy 988:1117 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 16 theme P0 976:977 arg1 protein 979:985 P0 protein 976:985 P0 protein 976:985 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 4 17 theme myelin 474:479 arg1 structures 481:490 abnormal myelin structures 465:490 abnormal myelin structures in many cases 465:504 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 9 18 dep PNS 1022:1024 arg1 myelin 1026:1031 myelin 1026:1031 myelin 1026:1031 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 6 19 theme system 662:667 arg1 myelination 675:685 peripheral nervous system (PNS) myelination 643:685 peripheral nervous system (PNS) myelination 643:685 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 11 20 theme axonal 1448:1453 arg1 survival 1455:1462 myelinated axonal survival 1437:1462 myelinated axonal survival 1437:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 11 20 theme axonal 1448:1453 arg1 PNS 1417:1419 PNS myelination and myelinated axonal survival 1417:1462 PNS myelination and myelinated axonal survival 1417:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 4 21 theme many 495:498 arg1 cases 500:504 many cases 495:504 many cases 495:504 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 3 22 theme Axonal 261:266 arg1 degeneration 268:279 Axonal degeneration 261:279 Axonal degeneration 261:279 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 3 22 theme Axonal 261:266 arg1 cause 294:298 the major cause 284:298 the major cause of permanent neurological disability in primary myelin diseases 284:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 11 23 theme N-glycans 1500:1508 arg1 GlcNAc-6-O-sulfation 1476:1495 the GlcNAc-6-O-sulfation 1472:1495 the GlcNAc-6-O-sulfation of N-glycans on glycoproteins 1472:1525 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 5 24 gly glycoproteins 547:559 arg1 glycoproteins 547:559 myelin glycoproteins 540:559 myelin glycoproteins 540:559 However, the roles of glycans on myelin glycoproteins remain poorly understood. 28186137 4 25 from structures 481:490 arg1 cases 500:504 many cases 495:504 many cases 495:504 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 0 26 theme peripheral 68:77 arg1 system 87:92 the peripheral nervous system 64:92 the peripheral nervous system 64:92 GlcNAc6ST-1 regulates sulfation of N-glycans and myelination in the peripheral nervous system. 28186137 9 27 gly glycosylation 1063:1075 arg2 site 1077:1080 the glycosylation site 1059:1080 the glycosylation site 1059:1080 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 5 28 from roles 520:524 arg1 glycoproteins 547:559 myelin glycoproteins 540:559 myelin glycoproteins 540:559 However, the roles of glycans on myelin glycoproteins remain poorly understood. 28186137 10 29 theme abnormal 1293:1300 arg1 myelination 1302:1312 abnormal myelination 1293:1312 abnormal myelination 1293:1312 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 10 30 theme deficient 1169:1177 arg1 Mice 1164:1167 Mice 1164:1167 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) 1164:1237 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 1 31 theme specialized 102:112 arg1 cells 120:124 Highly specialized glial cells 95:124 Highly specialized glial cells 95:124 Highly specialized glial cells wrap axons with a multilayered myelin membrane in vertebrates. 28186137 9 32 theme abundant 1124:1131 arg1 N-glycans 1153:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 33 theme abundant 997:1004 arg1 glycoprotein 1006:1017 the most abundant glycoprotein 988:1017 the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy 988:1117 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 33 theme abundant 997:1004 arg1 protein 979:985 P0 protein 976:985 P0 protein 976:985 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 3 34 theme major 288:292 arg1 degeneration 268:279 Axonal degeneration 261:279 Axonal degeneration 261:279 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 3 34 theme major 288:292 arg1 cause 294:298 the major cause 284:298 the major cause of permanent neurological disability in primary myelin diseases 284:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 7 35 theme myelin 692:697 arg1 glycoproteins 699:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 1 36 theme glial 114:118 arg1 cells 120:124 Highly specialized glial cells 95:124 Highly specialized glial cells 95:124 Highly specialized glial cells wrap axons with a multilayered myelin membrane in vertebrates. 28186137 9 37 theme GlcNAc-6-O-sulfated 1133:1151 arg1 N-glycans 1153:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 4 38 gly glycoproteins 370:382 arg1 glycoproteins 370:382 Many glycoproteins 365:382 Many glycoproteins 365:382 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 4 39 theme glycoprotein 441:452 arg1 lack 422:425 a lack 420:425 a lack of one myelin glycoprotein 420:452 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 0 40 theme N-glycans 35:43 arg1 sulfation 22:30 sulfation 22:30 sulfation of N-glycans and myelination 22:59 GlcNAc6ST-1 regulates sulfation of N-glycans and myelination in the peripheral nervous system. 28186137 8 41 theme N-acetylglucosamine 876:894 arg1 6-O-sulfation 859:871 the 6-O-sulfation 855:871 the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) 855:917 Major sulfated N-glycans were identified in both porcine and mouse PNS myelin, demonstrating that the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) is highly conserved in PNS myelin between these species. 28186137 4 42 theme abnormal 465:472 arg1 structures 481:490 abnormal myelin structures 465:490 abnormal myelin structures in many cases 465:504 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 7 43 contain contain 713:719 arg2 N-glycans 746:754 highly abundant sulfated N-glycans 721:754 highly abundant sulfated N-glycans 721:754 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 7 43 contain contain 713:719 arg1 glycoproteins 699:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 8 44 theme myelin 946:951 arg1 PNS 942:944 PNS 942:944 PNS myelin between these species 942:973 Major sulfated N-glycans were identified in both porcine and mouse PNS myelin, demonstrating that the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) is highly conserved in PNS myelin between these species. 28186137 12 45 theme novel 1555:1559 arg1 insights 1561:1568 novel insights 1555:1568 novel insights into the pathogenesis of peripheral neuropathy 1555:1615 These findings may provide novel insights into the pathogenesis of peripheral neuropathy. 28186137 11 46 theme myelinated 1437:1446 arg1 survival 1455:1462 myelinated axonal survival 1437:1462 myelinated axonal survival 1437:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 11 46 theme myelinated 1437:1446 arg1 PNS 1417:1419 PNS myelination and myelinated axonal survival 1417:1462 PNS myelination and myelinated axonal survival 1417:1462 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 3 47 theme permanent 303:311 arg1 disability 326:335 permanent neurological disability 303:335 permanent neurological disability 303:335 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 7 48 theme PNS 688:690 arg1 glycoproteins 699:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 5 49 theme myelin 540:545 arg1 glycoproteins 547:559 myelin glycoproteins 540:559 myelin glycoproteins 540:559 However, the roles of glycans on myelin glycoproteins remain poorly understood. 28186137 0 50 theme myelination 49:59 arg1 sulfation 22:30 sulfation 22:30 sulfation of N-glycans and myelination 22:59 GlcNAc6ST-1 regulates sulfation of N-glycans and myelination in the peripheral nervous system. 28186137 8 51 theme sulfated 763:770 arg1 N-glycans 772:780 Major sulfated N-glycans 757:780 Major sulfated N-glycans 757:780 Major sulfated N-glycans were identified in both porcine and mouse PNS myelin, demonstrating that the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) is highly conserved in PNS myelin between these species. 28186137 3 52 theme neurological 313:324 arg1 disability 326:335 permanent neurological disability 303:335 permanent neurological disability 303:335 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 12 53 theme peripheral 1595:1604 arg1 neuropathy 1606:1615 peripheral neuropathy 1595:1615 peripheral neuropathy 1595:1615 These findings may provide novel insights into the pathogenesis of peripheral neuropathy. 28186137 10 54 from N-acetylglucosamine-6-O-sulfotransferase-1 1182:1223 arg1 deficient 1169:1177 deficient 1169:1177 deficient 1169:1177 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 4 55 gly glycoprotein 441:452 arg1 glycoprotein 441:452 one myelin glycoprotein 430:452 one myelin glycoprotein 430:452 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 8 56 theme Major 757:761 arg1 N-glycans 772:780 Major sulfated N-glycans 757:780 Major sulfated N-glycans 757:780 Major sulfated N-glycans were identified in both porcine and mouse PNS myelin, demonstrating that the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) is highly conserved in PNS myelin between these species. 28186137 11 57 from GlcNAc-6-O-sulfation 1476:1495 arg1 glycoproteins 1513:1525 glycoproteins 1513:1525 glycoproteins 1513:1525 Taken together, this study demonstrates that GlcNAc6ST-1 modulates PNS myelination and myelinated axonal survival through the GlcNAc-6-O-sulfation of N-glycans on glycoproteins. 28186137 4 58 theme Many 365:368 arg1 glycoproteins 370:382 Many glycoproteins 365:382 Many glycoproteins 365:382 Many glycoproteins have been identified in myelin, and a lack of one myelin glycoprotein results in abnormal myelin structures in many cases. 28186137 6 59 theme nervous 654:660 arg1 PNS 670:672 PNS 670:672 PNS 670:672 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 6 59 theme nervous 654:660 arg1 system 662:667 peripheral nervous system 643:667 peripheral nervous system (PNS) myelination 643:685 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 3 60 from cause 294:298 arg1 diseases 355:362 primary myelin diseases 340:362 primary myelin diseases 340:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 2 61 theme essential 203:211 arg1 roles 213:217 essential roles 203:217 essential roles 203:217 Myelin serves essential roles in the functioning of the nervous system. 28186137 9 62 contain has 1120:1122 arg2 N-glycans 1153:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 abundant GlcNAc-6-O-sulfated N-glycans 1124:1161 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 62 contain has 1120:1122 arg1 glycoprotein 1006:1017 the most abundant glycoprotein 988:1017 the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy 988:1117 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 9 62 contain has 1120:1122 arg1 protein 979:985 P0 protein 976:985 P0 protein 976:985 P0 protein, the most abundant glycoprotein in PNS myelin and mutations in which at the glycosylation site cause Charcot-Marie-Tooth neuropathy, has abundant GlcNAc-6-O-sulfated N-glycans. 28186137 6 63 theme peripheral 643:652 arg1 PNS 670:672 PNS 670:672 PNS 670:672 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 6 63 theme peripheral 643:652 arg1 system 662:667 peripheral nervous system 643:667 peripheral nervous system (PNS) myelination 643:685 Here, we report that sulfated N-glycans are involved in peripheral nervous system (PNS) myelination. 28186137 3 64 theme primary 340:346 arg1 diseases 355:362 primary myelin diseases 340:362 primary myelin diseases 340:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 12 65 theme neuropathy 1606:1615 arg1 pathogenesis 1579:1590 the pathogenesis 1575:1590 the pathogenesis of peripheral neuropathy 1575:1615 These findings may provide novel insights into the pathogenesis of peripheral neuropathy. 28186137 10 66 theme axonal 1318:1323 arg1 degeneration 1325:1336 axonal degeneration 1318:1336 axonal degeneration 1318:1336 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 28186137 3 67 theme myelin 348:353 arg1 diseases 355:362 primary myelin diseases 340:362 primary myelin diseases 340:362 Axonal degeneration is the major cause of permanent neurological disability in primary myelin diseases. 28186137 2 68 theme nervous 245:251 arg1 system 253:258 the nervous system 241:258 the nervous system 241:258 Myelin serves essential roles in the functioning of the nervous system. 28186137 8 69 dep myelin 828:833 arg1 PNS 824:826 both porcine and mouse PNS myelin 801:833 PNS 824:826 Major sulfated N-glycans were identified in both porcine and mouse PNS myelin, demonstrating that the 6-O-sulfation of N-acetylglucosamine (GlcNAc-6-O-sulfation) is highly conserved in PNS myelin between these species. 28186137 1 70 theme multilayered 144:155 arg1 membrane 164:171 a multilayered myelin membrane 142:171 a multilayered myelin membrane in vertebrates 142:186 Highly specialized glial cells wrap axons with a multilayered myelin membrane in vertebrates. 28186137 7 71 gly glycoproteins 699:711 arg1 glycoproteins 699:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins 688:711 PNS myelin glycoproteins contain highly abundant sulfated N-glycans. 28186137 10 72 theme sulfated 1260:1267 arg1 N-glycans 1269:1277 sulfated N-glycans 1260:1277 sulfated N-glycans 1260:1277 Mice deficient in N-acetylglucosamine-6-O-sulfotransferase-1 (GlcNAc6ST-1) failed to synthesize sulfated N-glycans and exhibited abnormal myelination and axonal degeneration in the PNS. 26083631 0 0 from Glycan 12:17 arg1 Domain 29:34 the C2 Domain 22:34 the C2 Domain of HIV-1 Envelope 22:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 0 from Glycan 12:17 arg1 Envelope 45:52 HIV-1 Envelope 39:52 HIV-1 Envelope 39:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 7 1 theme divergent 882:890 arg1 use 898:900 divergent CXCR4 use 882:900 divergent CXCR4 use 882:900 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 1 2 theme CCR5-using 215:224 arg1 virus 226:230 exclusively CCR5-using virus 203:230 exclusively CCR5-using virus 203:230 Nearly all persons newly infected with HIV-1 harbor exclusively CCR5-using virus. 26083631 4 3 theme initial 443:449 arg1 dependence 451:460 HIV-1's initial dependence 435:460 HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, 435:522 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 0 4 theme V3 141:142 arg1 Loops 144:148 Identical V3 Loops 131:148 Identical V3 Loops 131:148 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 7 5 theme V3 869:870 arg1 loops 872:876 identical V3 loops 859:876 identical V3 loops 859:876 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 0 6 with Variants 117:124 arg1 Loops 144:148 Identical V3 Loops 131:148 Identical V3 Loops 131:148 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 7 theme Molecular 64:72 arg1 Glycan 12:17 A Conserved Glycan 0:17 A Conserved Glycan in the C2 Domain of HIV-1 Envelope 0:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 7 theme Molecular 64:72 arg1 Switch 74:79 a Molecular Switch 62:79 a Molecular Switch to Control X4 Utilization 62:105 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 4 8 from dependence 451:460 arg1 force 485:489 the selective force 471:489 the selective force(s) that drive CXCR4-utilization 471:521 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 4 8 from dependence 451:460 arg1 CCR5 465:468 CCR5 465:468 CCR5 465:468 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 5 9 theme Greater 602:608 arg1 knowledge 610:618 Greater knowledge 602:618 Greater knowledge of these processes 602:637 Greater knowledge of these processes will inform interventions at all stages, from vaccination to cure. 26083631 9 10 theme first 1068:1072 arg1 description 1074:1084 the first description 1064:1084 the first description of co-receptor usage regulated by the N-glycan at position 262 1064:1147 To our knowledge, this is the first description of co-receptor usage regulated by the N-glycan at position 262. 26083631 9 10 theme first 1068:1072 arg1 this 1056:1059 this 1056:1059 this 1056:1059 To our knowledge, this is the first description of co-receptor usage regulated by the N-glycan at position 262. 26083631 2 11 dep subtypes 310:317 arg1 B 319:319 B 319:319 B 319:319 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 2 11 dep subtypes 310:317 arg1 subtypes 310:317 subtypes B or D 310:324 subtypes B or D 310:324 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 2 11 dep subtypes 310:317 arg1 D 324:324 D 324:324 D 324:324 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 6 12 theme use 738:740 arg1 determinants 710:721 The determinants 706:721 The determinants of co-receptor use 706:740 The determinants of co-receptor use map primarily, though not exclusively, to the V3 loop of gp120. 26083631 8 13 dep residues 928:935 arg1 polymorphism 980:991 a rare polymorphism 973:991 a rare polymorphism in C1 973:997 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 13 dep residues 928:935 arg1 residues 928:935 two residues 924:935 two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2 924:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 13 dep residues 928:935 arg1 N-glycan 1022:1029 a highly conserved N-glycan 1003:1029 a highly conserved N-glycan in C2 1003:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 6 14 theme gp120 799:803 arg1 loop 791:794 the V3 loop 784:794 the V3 loop of gp120 784:803 The determinants of co-receptor use map primarily, though not exclusively, to the V3 loop of gp120. 26083631 8 15 from N-glycan 1022:1029 arg1 C1 996:997 C1 996:997 C1 996:997 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 15 from N-glycan 1022:1029 arg1 C2 1034:1035 C2 1034:1035 C2 1034:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 6 16 theme co-receptor 726:736 arg1 use 738:740 co-receptor use 726:740 co-receptor use 726:740 The determinants of co-receptor use map primarily, though not exclusively, to the V3 loop of gp120. 26083631 0 17 theme Conserved 2:10 arg1 Glycan 12:17 A Conserved Glycan 0:17 A Conserved Glycan in the C2 Domain of HIV-1 Envelope 0:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 17 theme Conserved 2:10 arg1 Switch 74:79 a Molecular Switch 62:79 a Molecular Switch to Control X4 Utilization 62:105 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 8 18 theme rare 975:978 arg1 polymorphism 980:991 a rare polymorphism 973:991 a rare polymorphism in C1 973:997 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 18 theme rare 975:978 arg1 residues 928:935 two residues 924:935 two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2 924:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 9 19 theme co-receptor 1089:1099 arg1 usage 1101:1105 co-receptor usage 1089:1105 co-receptor usage regulated by the N-glycan at position 262 1089:1147 To our knowledge, this is the first description of co-receptor usage regulated by the N-glycan at position 262. 26083631 0 20 theme Clonal 110:115 arg1 Variants 117:124 Clonal Variants 110:124 Clonal Variants with Identical V3 Loops 110:148 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 9 21 theme usage 1101:1105 arg1 description 1074:1084 the first description 1064:1084 the first description of co-receptor usage regulated by the N-glycan at position 262 1064:1147 To our knowledge, this is the first description of co-receptor usage regulated by the N-glycan at position 262. 26083631 9 21 theme usage 1101:1105 arg1 this 1056:1059 this 1056:1059 this 1056:1059 To our knowledge, this is the first description of co-receptor usage regulated by the N-glycan at position 262. 26083631 0 22 theme X4 92:93 arg1 Utilization 95:105 Control X4 Utilization 84:105 Control X4 Utilization 84:105 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 7 23 theme identical 859:867 arg1 loops 872:876 identical V3 loops 859:876 identical V3 loops 859:876 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 0 24 theme Control 84:90 arg1 Utilization 95:105 Control X4 Utilization 84:105 Control X4 Utilization 84:105 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 25 theme C2 26:27 arg1 Domain 29:34 the C2 Domain 22:34 the C2 Domain of HIV-1 Envelope 22:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 25 theme C2 26:27 arg1 Envelope 45:52 HIV-1 Envelope 39:52 HIV-1 Envelope 39:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 7 26 with variants 845:852 arg1 use 898:900 divergent CXCR4 use 882:900 divergent CXCR4 use 882:900 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 7 26 with variants 845:852 arg1 loops 872:876 identical V3 loops 859:876 identical V3 loops 859:876 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 6 27 theme V3 788:789 arg1 loop 791:794 the V3 loop 784:794 the V3 loop of gp120 784:803 The determinants of co-receptor use map primarily, though not exclusively, to the V3 loop of gp120. 26083631 3 28 theme efficient 346:354 arg1 utilization 362:372 efficient CXCR4 utilization 346:372 efficient CXCR4 utilization 346:372 This transition to efficient CXCR4 utilization is often co-incident with progression to AIDS. 26083631 3 29 theme CXCR4 356:360 arg1 utilization 362:372 efficient CXCR4 utilization 346:372 efficient CXCR4 utilization 346:372 This transition to efficient CXCR4 utilization is often co-incident with progression to AIDS. 26083631 3 30 with co-incident 383:393 arg1 progression 400:410 progression 400:410 progression to AIDS 400:418 This transition to efficient CXCR4 utilization is often co-incident with progression to AIDS. 26083631 8 31 theme phenotypic 954:963 arg1 switch 965:970 this phenotypic switch 949:970 this phenotypic switch 949:970 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 4 32 theme selective 475:483 arg1 s 491:491 s 491:491 s 491:491 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 4 32 theme selective 475:483 arg1 force 485:489 the selective force 471:489 the selective force(s) that drive CXCR4-utilization 471:521 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 4 32 theme selective 475:483 arg1 CCR5 465:468 CCR5 465:468 CCR5 465:468 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 0 33 theme Identical 131:139 arg1 Loops 144:148 Identical V3 Loops 131:148 Identical V3 Loops 131:148 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 7 34 theme clonal 838:843 arg1 variants 845:852 five clonal variants 833:852 five clonal variants with identical V3 loops but divergent CXCR4 use 833:900 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 2 35 theme CXCR4-using 233:243 arg1 variants 245:252 CXCR4-using variants 233:252 CXCR4-using variants 233:252 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 8 36 theme conserved 1012:1020 arg1 residues 928:935 two residues 924:935 two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2 924:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 36 theme conserved 1012:1020 arg1 N-glycan 1022:1029 a highly conserved N-glycan 1003:1029 a highly conserved N-glycan in C2 1003:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 2 37 theme patients 287:294 arg1 patients 287:294 patients 287:294 patients infected with subtypes B or D 287:324 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 2 37 theme patients 287:294 arg1 % 282:282 up to 50% 274:282 up to 50% of patients infected with subtypes B or D 274:324 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 4 38 theme evolutionary 532:543 arg1 pathways 545:552 the evolutionary pathways 528:552 the evolutionary pathways by which it occurs 528:571 The basis for HIV-1's initial dependence on CCR5, the selective force(s) that drive CXCR4-utilization, and the evolutionary pathways by which it occurs are incompletely understood. 26083631 5 39 theme processes 629:637 arg1 knowledge 610:618 Greater knowledge 602:618 Greater knowledge of these processes 602:637 Greater knowledge of these processes will inform interventions at all stages, from vaccination to cure. 26083631 7 40 theme CXCR4 892:896 arg1 use 898:900 divergent CXCR4 use 882:900 divergent CXCR4 use 882:900 In this study, we describe five clonal variants with identical V3 loops but divergent CXCR4 use. 26083631 2 41 dep 50 280:281 arg1 to 277:278 to 277:278 to 277:278 CXCR4-using variants eventually arise in up to 50% of patients infected with subtypes B or D. 26083631 0 42 theme Envelope 45:52 arg1 Domain 29:34 the C2 Domain 22:34 the C2 Domain of HIV-1 Envelope 22:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 42 theme Envelope 45:52 arg1 Envelope 45:52 HIV-1 Envelope 39:52 HIV-1 Envelope 39:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 0 43 theme HIV-1 39:43 arg1 Envelope 45:52 HIV-1 Envelope 39:52 HIV-1 Envelope 39:52 A Conserved Glycan in the C2 Domain of HIV-1 Envelope Acts as a Molecular Switch to Control X4 Utilization by Clonal Variants with Identical V3 Loops. 26083631 8 44 from polymorphism 980:991 arg1 C1 996:997 C1 996:997 C1 996:997 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 26083631 8 44 from polymorphism 980:991 arg1 C2 1034:1035 C2 1034:1035 C2 1034:1035 Mutagenesis revealed two residues controlling this phenotypic switch: a rare polymorphism in C1 and a highly conserved N-glycan in C2. 25546301 0 0 theme stabilizing 69:79 arg1 role 81:84 a stabilizing role 67:84 a stabilizing role for the glycan at Asn262 67:109 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 1 1 theme gp120 281:285 arg1 glycans 265:271 the 15 N-linked glycans 249:271 the 15 N-linked glycans of core gp120 249:285 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 2 gly glycosylated 145:156 arg1 core 170:173 a fully glycosylated HIV-1 gp120 core 137:173 a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b 137:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 3 3 dep gp120 526:530 arg1 stability 544:552 stability 544:552 stability 544:552 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 3 3 dep gp120 526:530 arg1 folding 532:538 folding 532:538 folding 532:538 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 4 4 from patch 633:637 arg1 glycans 603:609 neighboring glycans 591:609 neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies 591:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 4 4 from patch 633:637 arg1 stems 582:586 the stems 578:586 the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies 578:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 4 5 theme oligomannose 620:631 arg1 patch 633:637 the oligomannose patch 616:637 the oligomannose patch 616:637 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 4 5 theme oligomannose 620:631 arg1 target 657:662 a major target 649:662 a major target for broadly neutralizing antibodies 649:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 1 6 theme 4.5-Å 219:223 arg1 resolution 225:234 4.5-Å resolution 219:234 4.5-Å resolution 219:234 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 3 7 theme GlcNAc 439:444 arg1 stem 446:449 The GlcNAc stem 435:449 The GlcNAc stem of this glycan 435:464 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 1 8 from structure 124:132 arg1 complex 178:184 complex 178:184 complex with CD4 receptor and Fab 17b 178:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 3 9 from cleft 489:493 arg1 gp120 498:502 gp120 498:502 gp120 498:502 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 3 10 from role 518:521 arg1 gp120 526:530 gp120 folding and stability 526:552 gp120 folding and stability 526:552 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 2 11 theme GlcNAc 406:411 arg1 7 419:419 7 419:419 7 419:419 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 2 11 theme GlcNAc 406:411 arg1 Man 415:417 a GlcNAc(2)Man 404:417 a GlcNAc(2)Man(7) 404:420 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 2 12 theme electron 382:389 arg1 density 391:397 the most extensive and well-ordered electron density 346:397 the most extensive and well-ordered electron density 346:397 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 1 13 theme glycosylated 145:156 arg1 core 170:173 a fully glycosylated HIV-1 gp120 core 137:173 a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b 137:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 4 14 theme neighboring 591:601 arg1 glycans 603:609 neighboring glycans 591:609 neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies 591:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 1 15 theme HIV-1 158:162 arg1 core 170:173 a fully glycosylated HIV-1 gp120 core 137:173 a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b 137:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 0 16 theme Crystal 0:6 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a fully glycosylated HIV-1 gp120 core 0:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 3 17 theme glycan 459:464 arg1 stem 446:449 The GlcNAc stem 435:449 The GlcNAc stem of this glycan 435:464 The GlcNAc stem of this glycan is largely buried in a cleft in gp120, suggesting a role in gp120 folding and stability. 25546301 1 18 theme gp120 164:168 arg1 core 170:173 a fully glycosylated HIV-1 gp120 core 137:173 a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b 137:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 2 19 theme well-ordered 369:380 arg1 density 391:397 the most extensive and well-ordered electron density 346:397 the most extensive and well-ordered electron density 346:397 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 1 20 with complex 178:184 arg1 receptor 195:202 CD4 receptor 191:202 CD4 receptor 191:202 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 20 with complex 178:184 arg1 17b 212:214 Fab 17b 208:214 Fab 17b 208:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 21 theme core 170:173 arg1 structure 124:132 The crystal structure 112:132 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution 112:234 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 22 from resolution 225:234 arg1 structure 124:132 The crystal structure 112:132 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution 112:234 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 4 23 theme glycans 603:609 arg1 stems 582:586 the stems 578:586 the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies 578:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 0 24 theme glycosylated 29:40 arg1 core 54:57 a fully glycosylated HIV-1 gp120 core 21:57 a fully glycosylated HIV-1 gp120 core 21:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 1 25 from complex 178:184 arg1 structure 124:132 The crystal structure 112:132 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution 112:234 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 0 26 theme gp120 48:52 arg1 core 54:57 a fully glycosylated HIV-1 gp120 core 21:57 a fully glycosylated HIV-1 gp120 core 21:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 1 27 theme N-linked 256:263 arg1 glycans 265:271 the 15 N-linked glycans 249:271 the 15 N-linked glycans of core gp120 249:285 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 4 28 theme neutralizing 676:687 arg1 antibodies 689:698 broadly neutralizing antibodies 668:698 broadly neutralizing antibodies 668:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 0 29 theme HIV-1 42:46 arg1 core 54:57 a fully glycosylated HIV-1 gp120 core 21:57 a fully glycosylated HIV-1 gp120 core 21:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 1 30 theme glycans 265:271 arg1 glycans 265:271 the 15 N-linked glycans 249:271 the 15 N-linked glycans of core gp120 249:285 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 30 theme glycans 265:271 arg1 9 244:244 9 244:244 9 244:244 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 2 31 contain had 342:344 arg1 glycan 316:321 The glycan 312:321 The glycan at position Asn262 312:340 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 2 31 contain had 342:344 arg2 density 391:397 the most extensive and well-ordered electron density 346:397 the most extensive and well-ordered electron density 346:397 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 1 32 from core 170:173 arg1 complex 178:184 complex 178:184 complex with CD4 receptor and Fab 17b 178:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 4 33 from stems 582:586 arg1 patch 633:637 the oligomannose patch 616:637 the oligomannose patch 616:637 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 4 33 from stems 582:586 arg1 target 657:662 a major target 649:662 a major target for broadly neutralizing antibodies 649:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 1 34 link N-linked 256:263 arg1 glycans 265:271 the 15 N-linked glycans 249:271 the 15 N-linked glycans of core gp120 249:285 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 35 theme CD4 191:193 arg1 receptor 195:202 CD4 receptor 191:202 CD4 receptor 191:202 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 2 36 from position 326:333 arg1 glycan 316:321 The glycan 312:321 The glycan at position Asn262 312:340 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 0 37 gly glycosylated 29:40 arg1 core 54:57 a fully glycosylated HIV-1 gp120 core 21:57 a fully glycosylated HIV-1 gp120 core 21:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 4 38 theme major 651:655 arg1 patch 633:637 the oligomannose patch 616:637 the oligomannose patch 616:637 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 4 38 theme major 651:655 arg1 target 657:662 a major target 649:662 a major target for broadly neutralizing antibodies 649:698 Its arms interact with the stems of neighboring glycans from the oligomannose patch, which is a major target for broadly neutralizing antibodies. 25546301 2 39 theme extensive 355:363 arg1 density 391:397 the most extensive and well-ordered electron density 346:397 the most extensive and well-ordered electron density 346:397 The glycan at position Asn262 had the most extensive and well-ordered electron density, and a GlcNAc(2)Man(7) was modeled. 25546301 1 40 theme crystal 116:122 arg1 structure 124:132 The crystal structure 112:132 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution 112:234 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 0 41 from Asn262 104:109 arg1 glycan 94:99 the glycan 90:99 the glycan at Asn262 90:109 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 0 42 theme core 54:57 arg1 structure 8:16 Crystal structure 0:16 Crystal structure of a fully glycosylated HIV-1 gp120 core 0:57 Crystal structure of a fully glycosylated HIV-1 gp120 core reveals a stabilizing role for the glycan at Asn262. 25546301 1 43 theme core 276:279 arg1 gp120 281:285 core gp120 276:285 core gp120 276:285 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 25546301 1 44 theme Fab 208:210 arg1 17b 212:214 Fab 17b 208:214 Fab 17b 208:214 The crystal structure of a fully glycosylated HIV-1 gp120 core in complex with CD4 receptor and Fab 17b at 4.5-Å resolution reveals 9 of the 15 N-linked glycans of core gp120 to be partially ordered. 24522230 4 0 theme many 473:476 arg1 cases 478:482 many cases 473:482 many cases 473:482 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 9 1 from those 1211:1215 arg1 distinct 1197:1204 distinct 1197:1204 distinct 1197:1204 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 4 2 from shifts 587:592 arg1 sampling 615:622 Fc conformational sampling 597:622 Fc conformational sampling 597:622 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 6 3 theme motion 825:830 arg1 amplitude 812:820 amplitude 812:820 amplitude of motion for the N-terminal Cγ2 domains 812:861 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 6 3 theme motion 825:830 arg1 N-glycan 867:874 N-glycan 867:874 N-glycan 867:874 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 8 4 theme Fc 1053:1054 arg1 model 1044:1048 a model 1042:1048 a model of Fc that is structurally dynamic 1042:1083 Our results are consistent with a model of Fc that is structurally dynamic. 24522230 8 4 theme Fc 1053:1054 arg1 dynamic 1077:1083 dynamic 1077:1083 dynamic 1077:1083 Our results are consistent with a model of Fc that is structurally dynamic. 24522230 1 5 dep identification 133:146 arg1 the 116:118 the 116:118 the 116:118 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 9 6 theme FcγR 1229:1232 arg1 complexes 1234:1242 FcγR complexes 1229:1242 FcγR complexes 1229:1242 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 9 7 theme Conformational 1086:1099 arg1 states 1101:1106 Conformational states 1086:1106 Conformational states that are competent to bind immune-stimulating FcγRs 1086:1158 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 1 8 theme destruction 152:162 arg1 properties 164:173 destruction properties 152:173 destruction properties 152:173 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 3 9 theme committal 332:340 arg1 response 383:390 committal to a damaging, though protective, immune response 332:390 committal to a damaging, though protective, immune response 332:390 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 2 10 theme Fcγ 267:269 arg1 FcγRs 282:286 FcγRs 282:286 FcγRs 282:286 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 2 10 theme Fcγ 267:269 arg1 receptors 271:279 pro-inflammatory Fcγ receptors 250:279 pro-inflammatory Fcγ receptors (FcγRs) on immune cells 250:303 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 3 11 theme multiple 418:425 arg1 levels 427:432 multiple levels 418:432 multiple levels 418:432 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 0 12 theme G1 15:16 arg1 motions 28:34 Immunoglobulin G1 Fc domain motions 0:34 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 0:68 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 1 13 theme fragment 74:81 arg1 region 103:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 5 14 theme IgG1 661:664 arg1 conformation 669:680 a previously unseen IgG1 Fc conformation 641:680 a previously unseen IgG1 Fc conformation 641:680 Here, we report a previously unseen IgG1 Fc conformation. 24522230 0 15 theme Immunoglobulin 0:13 arg1 motions 28:34 Immunoglobulin G1 Fc domain motions 0:34 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 0:68 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 6 16 theme motions 782:788 arg1 investigation 742:754 an extensive molecular dynamics investigation 710:754 an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed 710:899 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 2 17 theme immune 292:297 arg1 cells 299:303 immune cells 292:303 immune cells 292:303 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 1 18 theme immunoglobulin 178:191 arg1 IgG 196:198 IgG 196:198 IgG 196:198 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 1 18 theme immunoglobulin 178:191 arg1 G 193:193 immunoglobulin G 178:193 immunoglobulin G (IgG) 178:199 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 0 19 theme domain 21:26 arg1 motions 28:34 Immunoglobulin G1 Fc domain motions 0:34 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 0:68 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 9 20 theme Fc 1180:1181 arg1 conformations 1183:1195 Fc conformations 1180:1195 Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population 1180:1298 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 6 21 theme glycan 775:780 arg1 motions 782:788 polypeptide and glycan motions 759:788 motions 782:788 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 6 22 dep greater 804:810 arg1 amplitude 812:820 amplitude 812:820 amplitude of motion for the N-terminal Cγ2 domains 812:861 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 6 22 dep greater 804:810 arg1 N-glycan 867:874 N-glycan 867:874 N-glycan 867:874 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 1 23 theme G 193:193 arg1 identification 133:146 key pathogen identification 120:146 key pathogen identification 120:146 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 1 23 theme G 193:193 arg1 properties 164:173 destruction properties 152:173 destruction properties 152:173 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 1 24 theme crystallizable 83:96 arg1 region 103:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 4 25 theme FcγR 549:552 arg1 affinity 554:561 FcγR affinity 549:561 FcγR affinity being modulated through shifts in Fc conformational sampling 549:622 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 1 26 theme Fc 99:100 arg1 region 103:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region 70:108 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 7 27 theme Cγ2/Cγ3 918:924 arg1 interface 926:934 the Cγ2/Cγ3 interface 914:934 the Cγ2/Cγ3 interface 914:934 Residues in the Cγ2/Cγ3 interface and disulfide-bonded hinge were identified as influencing the Cγ2 motion. 24522230 9 28 theme transient 1267:1275 arg1 population 1289:1298 a transient, nonbinding population 1265:1298 a transient, nonbinding population 1265:1298 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 9 29 dep transient 1267:1275 arg1 nonbinding 1278:1287 nonbinding 1278:1287 nonbinding 1278:1287 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 9 30 theme immune-stimulating 1135:1152 arg1 FcγRs 1154:1158 immune-stimulating FcγRs 1135:1158 immune-stimulating FcγRs 1135:1158 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 5 31 theme unseen 654:659 arg1 conformation 669:680 a previously unseen IgG1 Fc conformation 641:680 a previously unseen IgG1 Fc conformation 641:680 Here, we report a previously unseen IgG1 Fc conformation. 24522230 4 32 theme Control 435:441 arg1 mechanisms 443:452 Control mechanisms 435:452 Control mechanisms 435:452 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 2 33 theme Pathogen 202:209 arg1 opsonization 211:222 Pathogen opsonization 202:222 Pathogen opsonization 202:222 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 6 34 theme polypeptide 759:769 arg1 motions 782:788 polypeptide and glycan motions 759:788 motions 782:788 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 3 35 theme immune 376:381 arg1 response 383:390 committal to a damaging, though protective, immune response 332:390 committal to a damaging, though protective, immune response 332:390 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 4 36 theme suggested 512:520 arg1 contribution 522:533 one frequently suggested contribution 497:533 one frequently suggested contribution 497:533 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 3 37 dep committal 332:340 arg1 though 357:362 though 357:362 though 357:362 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 3 37 dep committal 332:340 arg1 protective 364:373 protective 364:373 protective 364:373 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 6 38 theme Cγ2 851:853 arg1 domains 855:861 the N-terminal Cγ2 domains 836:861 the N-terminal Cγ2 domains 836:861 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 0 39 dep motions 28:34 arg1 implications 37:48 implications 37:48 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 0:68 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 2 40 from receptors 271:279 arg1 cells 299:303 immune cells 292:303 immune cells 292:303 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 7 41 theme disulfide-bonded 940:955 arg1 hinge 957:961 disulfide-bonded hinge 940:961 disulfide-bonded hinge 940:961 Residues in the Cγ2/Cγ3 interface and disulfide-bonded hinge were identified as influencing the Cγ2 motion. 24522230 5 42 theme Fc 666:667 arg1 conformation 669:680 a previously unseen IgG1 Fc conformation 641:680 a previously unseen IgG1 Fc conformation 641:680 Here, we report a previously unseen IgG1 Fc conformation. 24522230 6 43 theme N-terminal 840:849 arg1 domains 855:861 the N-terminal Cγ2 domains 836:861 the N-terminal Cγ2 domains 836:861 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 2 44 theme pro-inflammatory 250:265 arg1 FcγRs 282:286 FcγRs 282:286 FcγRs 282:286 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 2 44 theme pro-inflammatory 250:265 arg1 receptors 271:279 pro-inflammatory Fcγ receptors 250:279 pro-inflammatory Fcγ receptors (FcγRs) on immune cells 250:303 Pathogen opsonization positions Fcs to activate pro-inflammatory Fcγ receptors (FcγRs) on immune cells. 24522230 7 45 theme Cγ2 998:1000 arg1 motion 1002:1007 the Cγ2 motion 994:1007 the Cγ2 motion 994:1007 Residues in the Cγ2/Cγ3 interface and disulfide-bonded hinge were identified as influencing the Cγ2 motion. 24522230 9 46 located observed 1217:1224 arg2 those 1211:1215 those 1211:1215 those 1211:1215 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 9 46 located observed 1217:1224 arg1 complexes 1234:1242 FcγR complexes 1229:1242 FcγR complexes 1229:1242 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 7 47 from Residues 902:909 arg1 hinge 957:961 disulfide-bonded hinge 940:961 disulfide-bonded hinge 940:961 Residues in the Cγ2/Cγ3 interface and disulfide-bonded hinge were identified as influencing the Cγ2 motion. 24522230 7 47 from Residues 902:909 arg1 interface 926:934 the Cγ2/Cγ3 interface 914:934 the Cγ2/Cγ3 interface 914:934 Residues in the Cγ2/Cγ3 interface and disulfide-bonded hinge were identified as influencing the Cγ2 motion. 24522230 4 48 dep diverse 458:464 arg1 unclear 484:490 unclear 484:490 unclear 484:490 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 9 49 theme distinct 1197:1204 arg1 conformations 1183:1195 Fc conformations 1180:1195 Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population 1180:1298 Conformational states that are competent to bind immune-stimulating FcγRs interconverted with Fc conformations distinct from those observed in FcγR complexes, which may represent a transient, nonbinding population. 24522230 6 50 theme dynamics 733:740 arg1 investigation 742:754 an extensive molecular dynamics investigation 710:754 an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed 710:899 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 3 51 theme cellular 310:317 arg1 response 319:326 The cellular response 306:326 The cellular response 306:326 The cellular response and committal to a damaging, though protective, immune response are tightly controlled at multiple levels. 24522230 8 52 with consistent 1026:1035 arg1 model 1044:1048 a model 1042:1048 a model of Fc that is structurally dynamic 1042:1083 Our results are consistent with a model of Fc that is structurally dynamic. 24522230 8 52 with consistent 1026:1035 arg1 dynamic 1077:1083 dynamic 1077:1083 dynamic 1077:1083 Our results are consistent with a model of Fc that is structurally dynamic. 24522230 6 53 theme molecular 723:731 arg1 investigation 742:754 an extensive molecular dynamics investigation 710:754 an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed 710:899 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 1 54 theme key 120:122 arg1 identification 133:146 key pathogen identification 120:146 key pathogen identification 120:146 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 4 55 theme conformational 600:613 arg1 sampling 615:622 Fc conformational sampling 597:622 Fc conformational sampling 597:622 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 24522230 0 56 theme Fc 54:55 arg1 engineering 57:67 Fc engineering 54:67 Fc engineering 54:67 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 0 57 theme Fc 18:19 arg1 motions 28:34 Immunoglobulin G1 Fc domain motions 0:34 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 0:68 Immunoglobulin G1 Fc domain motions: implications for Fc engineering. 24522230 6 58 theme extensive 713:721 arg1 investigation 742:754 an extensive molecular dynamics investigation 710:754 an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed 710:899 This observation motivated an extensive molecular dynamics investigation of polypeptide and glycan motions that revealed greater amplitude of motion for the N-terminal Cγ2 domains and N-glycan than previously observed. 24522230 1 59 theme pathogen 124:131 arg1 identification 133:146 key pathogen identification 120:146 key pathogen identification 120:146 The fragment crystallizable (Fc) region links the key pathogen identification and destruction properties of immunoglobulin G (IgG). 24522230 4 60 theme Fc 597:598 arg1 sampling 615:622 Fc conformational sampling 597:622 Fc conformational sampling 597:622 Control mechanisms are diverse and in many cases unclear, but one frequently suggested contribution originates in FcγR affinity being modulated through shifts in Fc conformational sampling. 26940363 2 0 dep information 583:593 arg1 ii 579:580 ii 579:580 ii 579:580 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 1 theme glycan 623:628 arg1 heterogeneity 630:642 site-specific glycan heterogeneity 609:642 site-specific glycan heterogeneity 609:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 1 2 theme curated 262:268 arg1 database 270:277 a curated database 260:277 a curated database 260:277 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 6 3 theme glycan 1191:1196 arg1 data 1208:1211 query relative glycan abundance data 1176:1211 query relative glycan abundance data 1176:1211 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 11 4 theme disease-related 2115:2129 arg1 advancements 2042:2053 Continuing advancements 2031:2053 Continuing advancements in analytical technologies and new data types 2031:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 11 4 theme disease-related 2115:2129 arg1 glycomics 2131:2139 advancing disease-related glycomics 2105:2139 advancing disease-related glycomics 2105:2139 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 6 5 theme query 1176:1180 arg1 data 1208:1211 query relative glycan abundance data 1176:1211 query relative glycan abundance data 1176:1211 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 6 theme cancer 1262:1267 arg1 cancer 1262:1267 prostate cancer 1253:1267 prostate cancer 1253:1267 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 6 theme cancer 1262:1267 arg1 examples 1283:1290 examples 1283:1290 examples 1283:1290 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 6 theme cancer 1262:1267 arg1 set 1221:1223 a set 1219:1223 a set of published colorectal and prostate cancer cell lines 1219:1278 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 6 theme cancer 1262:1267 arg1 colorectal 1238:1247 colorectal 1238:1247 colorectal 1238:1247 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 4 7 theme data 872:875 arg1 model 877:881 our data model 868:881 our data model 868:881 METHODS: Since the launch of UniCarbKB in 2012, we have continued to improve the organisation of our data model. 26940363 8 8 from heterogeneity 1601:1613 arg1 representation 1536:1549 the efficient representation 1522:1549 the efficient representation of disease terms and associated changes in glycan heterogeneity 1522:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 8 9 from changes 1583:1589 arg1 heterogeneity 1601:1613 glycan heterogeneity 1594:1613 glycan heterogeneity 1594:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 10 10 theme site-specific 1832:1844 arg1 data 1860:1863 site-specific glycosylation data 1832:1863 site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies 1832:1943 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 4 11 theme model 877:881 arg1 organisation 852:863 the organisation 848:863 the organisation of our data model 848:881 METHODS: Since the launch of UniCarbKB in 2012, we have continued to improve the organisation of our data model. 26940363 6 12 theme UniCarbKB 1153:1161 arg1 capability 1139:1148 the capability 1135:1148 the capability of UniCarbKB to store and query relative glycan abundance data 1135:1211 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 11 13 from advancements 2042:2053 arg1 types 2095:2099 new data types 2086:2099 new data types 2086:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 11 13 from advancements 2042:2053 arg1 technologies 2069:2080 analytical technologies 2058:2080 analytical technologies 2058:2080 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 0 14 theme disease 117:123 arg1 associations 125:136 disease associations 117:136 disease associations 117:136 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 5 15 theme glycome 1082:1088 arg1 coverage 1060:1067 our coverage 1056:1067 our coverage of the human glycome 1056:1088 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 2 16 theme purified 551:558 arg1 glycoproteins 560:572 single purified glycoproteins 544:572 single purified glycoproteins 544:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 7 17 theme glycan 1395:1400 arg1 data 1416:1419 glycan compositional data 1395:1419 glycan compositional data 1395:1419 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 11 18 theme analytical 2058:2067 arg1 technologies 2069:2080 analytical technologies 2058:2080 analytical technologies 2058:2080 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 9 19 gly glycoproteins 1798:1810 arg1 glycoproteins 1798:1810 mammalian glycoproteins 1788:1810 mammalian glycoproteins 1788:1810 CONCLUSIONS: Updates and improvements to UniCarbKB have introduced unique features for storing and displaying glycosylation features of mammalian glycoproteins. 26940363 1 20 theme mammalian 214:222 arg1 knowledge 237:245 mammalian glycobiology knowledge 214:245 mammalian glycobiology knowledge 214:245 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 2 21 theme information 583:593 arg1 levels 446:451 two levels 442:451 two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity 442:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 10 22 theme data 1860:1863 arg1 integration 1817:1827 The integration 1813:1827 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies 1813:1943 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 11 23 theme new 2086:2088 arg1 types 2095:2099 new data types 2086:2099 new data types 2086:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 10 24 theme introduction 1911:1922 arg1 studies 1937:1943 large-scale glycoproteomics and introduction of cell line studies 1879:1943 large-scale glycoproteomics and introduction of cell line studies 1879:1943 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 5 25 theme structural 935:944 arg1 changes 960:966 structural and abundance changes 935:966 changes 960:966 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 9 26 theme mammalian 1788:1796 arg1 glycoproteins 1798:1810 mammalian glycoproteins 1788:1810 mammalian glycoproteins 1788:1810 CONCLUSIONS: Updates and improvements to UniCarbKB have introduced unique features for storing and displaying glycosylation features of mammalian glycoproteins. 26940363 7 27 theme large-scale 1343:1353 arg1 data 1371:1374 large-scale glycoproteomics data 1343:1374 large-scale glycoproteomics data 1343:1374 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 0 28 theme structure 56:64 arg1 abundance 66:74 glycan structure abundance 49:74 glycan structure abundance 49:74 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 1 29 theme web 331:333 arg1 application 335:345 a web application 329:345 a web application that allows users to easily find and view richly annotated information 329:416 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 10 30 theme glycoproteomics 1891:1905 arg1 studies 1937:1943 large-scale glycoproteomics and introduction of cell line studies 1879:1943 large-scale glycoproteomics and introduction of cell line studies 1879:1943 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 13 31 theme data 2344:2347 arg1 range 2335:2339 a wide range 2328:2339 a wide range of data with corresponding annotations 2328:2378 UniCarbKB was established with the mission of providing a resource for human glycobiology by capturing a wide range of data with corresponding annotations. 26940363 6 32 dep colorectal 1238:1247 arg1 lines 1274:1278 cell lines 1269:1278 cell lines 1269:1278 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 14 33 theme Issue 2415:2419 arg1 part 2397:2400 part 2397:2400 part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc 2397:2499 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 2 34 dep annotation 456:465 arg1 i 468:468 i 468:468 i 468:468 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 34 dep annotation 456:465 arg1 data 487:490 global-specific data 471:490 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 35 from levels 446:451 arg1 glycoproteins 560:572 single purified glycoproteins 544:572 single purified glycoproteins 544:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 0 36 theme glycoproteomics 91:105 arg1 data 107:110 compositional glycoproteomics data 77:110 compositional glycoproteomics data 77:110 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 6 37 theme relative 1182:1189 arg1 data 1208:1211 query relative glycan abundance data 1176:1211 query relative glycan abundance data 1176:1211 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 5 38 theme human 1001:1005 arg1 states 1015:1020 different human disease states 991:1020 different human disease states 991:1020 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 2 39 theme oligosaccharides 495:510 arg1 annotation 456:465 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 39 theme oligosaccharides 495:510 arg1 information 583:593 information 583:593 (ii) information pertaining to site-specific glycan heterogeneity 578:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 1 40 theme structural 282:291 arg1 data 310:313 structural and experimental data 282:313 structural and experimental data 282:313 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 5 41 theme abundance 950:958 arg1 changes 960:966 structural and abundance changes 935:966 changes 960:966 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 8 42 theme disease 1554:1560 arg1 terms 1562:1566 disease terms 1554:1566 disease terms 1554:1566 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 0 43 theme New 11:13 arg1 features 24:31 New database features 11:31 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 0:137 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 1 44 theme experimental 297:308 arg1 data 310:313 structural and experimental data 282:313 structural and experimental data 282:313 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 13 45 theme corresponding 2354:2366 arg1 annotations 2368:2378 corresponding annotations 2354:2378 corresponding annotations 2354:2378 UniCarbKB was established with the mission of providing a resource for human glycobiology by capturing a wide range of data with corresponding annotations. 26940363 8 46 from representation 1536:1549 arg1 heterogeneity 1601:1613 glycan heterogeneity 1594:1613 glycan heterogeneity 1594:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 8 47 theme changes 1583:1589 arg1 representation 1536:1549 the efficient representation 1522:1549 the efficient representation of disease terms and associated changes in glycan heterogeneity 1522:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 10 48 theme cell 1927:1930 arg1 line 1932:1935 cell line 1927:1935 cell line 1927:1935 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 6 49 theme abundance 1198:1206 arg1 data 1208:1211 query relative glycan abundance data 1176:1211 query relative glycan abundance data 1176:1211 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 3 50 theme bibliographic 714:726 arg1 information 743:753 structural, bibliographic, and taxonomic information 702:753 structural, bibliographic, and taxonomic information for each entry 702:768 Additional, contextual information is provided including structural, bibliographic, and taxonomic information for each entry. 26940363 1 51 theme glycobiology 224:235 arg1 knowledge 237:245 mammalian glycobiology knowledge 214:245 mammalian glycobiology knowledge 214:245 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 3 52 theme structural 702:711 arg1 information 743:753 structural, bibliographic, and taxonomic information 702:753 structural, bibliographic, and taxonomic information for each entry 702:768 Additional, contextual information is provided including structural, bibliographic, and taxonomic information for each entry. 26940363 13 53 theme human 2296:2300 arg1 glycobiology 2302:2313 human glycobiology 2296:2313 human glycobiology 2296:2313 UniCarbKB was established with the mission of providing a resource for human glycobiology by capturing a wide range of data with corresponding annotations. 26940363 5 54 theme experimental 1026:1037 arg1 models 1039:1044 experimental models 1026:1044 experimental models 1026:1044 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 14 55 dep Glycans 2431:2437 arg1 Lauc 2496:2499 Lauc 2496:2499 Lauc 2496:2499 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 2 56 from glycoproteins 560:572 arg1 levels 446:451 two levels 442:451 two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity 442:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 56 from glycoproteins 560:572 arg1 annotation 456:465 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 56 from glycoproteins 560:572 arg1 information 583:593 information 583:593 (ii) information pertaining to site-specific glycan heterogeneity 578:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 10 57 theme entire 1992:1997 arg1 glycomes 1999:2006 entire glycomes 1992:2006 entire glycomes 1992:2006 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 2 58 theme site-specific 609:621 arg1 heterogeneity 630:642 site-specific glycan heterogeneity 609:642 site-specific glycan heterogeneity 609:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 10 59 theme glycoproteins 1974:1986 arg1 analysis 1962:1969 the analysis 1958:1969 the analysis of glycoproteins and entire glycomes 1958:2006 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 8 60 from terms 1562:1566 arg1 heterogeneity 1601:1613 glycan heterogeneity 1594:1613 glycan heterogeneity 1594:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 11 61 theme advancing 2105:2113 arg1 advancements 2042:2053 Continuing advancements 2031:2053 Continuing advancements in analytical technologies and new data types 2031:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 11 61 theme advancing 2105:2113 arg1 glycomics 2131:2139 advancing disease-related glycomics 2105:2139 advancing disease-related glycomics 2105:2139 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 7 62 theme adding 1450:1455 arg1 value 1457:1461 adding value 1450:1461 adding value to UniCarbKB 1450:1474 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 6 63 theme cell 1269:1272 arg1 lines 1274:1278 cell lines 1269:1278 cell lines 1269:1278 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 13 64 with data 2344:2347 arg1 annotations 2368:2378 corresponding annotations 2354:2378 corresponding annotations 2354:2378 UniCarbKB was established with the mission of providing a resource for human glycobiology by capturing a wide range of data with corresponding annotations. 26940363 6 65 theme prostate 1253:1260 arg1 cancer 1262:1267 prostate cancer 1253:1267 prostate cancer 1253:1267 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 14 66 theme Guest 2465:2469 arg1 Editor 2471:2476 Guest Editor 2465:2476 Guest Editor 2465:2476 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 10 67 theme glycosylation 1846:1858 arg1 data 1860:1863 site-specific glycosylation data 1832:1863 site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies 1832:1943 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 9 68 theme unique 1719:1724 arg1 features 1726:1733 unique features 1719:1733 unique features 1719:1733 CONCLUSIONS: Updates and improvements to UniCarbKB have introduced unique features for storing and displaying glycosylation features of mammalian glycoproteins. 26940363 10 69 theme large-scale 1879:1889 arg1 glycoproteomics 1891:1905 large-scale glycoproteomics 1879:1905 large-scale glycoproteomics 1879:1905 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 3 70 dep Additional 645:654 arg1 contextual 657:666 contextual 657:666 contextual 657:666 Additional, contextual information is provided including structural, bibliographic, and taxonomic information for each entry. 26940363 2 71 theme single 544:549 arg1 glycoproteins 560:572 single purified glycoproteins 544:572 single purified glycoproteins 544:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 7 72 theme compositional 1402:1414 arg1 data 1416:1419 glycan compositional data 1395:1419 glycan compositional data 1395:1419 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 8 73 theme glycan 1594:1599 arg1 heterogeneity 1601:1613 glycan heterogeneity 1594:1613 glycan heterogeneity 1594:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 1 74 theme annotated 396:404 arg1 information 406:416 annotated information 396:416 annotated information 396:416 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 11 75 theme Continuing 2031:2040 arg1 advancements 2042:2053 Continuing advancements 2031:2053 Continuing advancements in analytical technologies and new data types 2031:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 11 75 theme Continuing 2031:2040 arg1 glycomics 2131:2139 advancing disease-related glycomics 2105:2139 advancing disease-related glycomics 2105:2139 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 9 76 theme glycosylation 1762:1774 arg1 features 1776:1783 glycosylation features 1762:1783 glycosylation features of mammalian glycoproteins 1762:1810 CONCLUSIONS: Updates and improvements to UniCarbKB have introduced unique features for storing and displaying glycosylation features of mammalian glycoproteins. 26940363 14 77 theme personalised 2442:2453 arg1 medicine 2455:2462 personalised medicine 2442:2462 personalised medicine" Guest Editor 2442:2476 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 3 78 theme Additional 645:654 arg1 information 668:678 Additional, contextual information 645:678 Additional, contextual information 645:678 Additional, contextual information is provided including structural, bibliographic, and taxonomic information for each entry. 26940363 5 79 theme human 1076:1080 arg1 glycome 1082:1088 the human glycome 1072:1088 the human glycome 1072:1088 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 2 80 gly glycoproteins 560:572 arg1 glycoproteins 560:572 single purified glycoproteins 544:572 single purified glycoproteins 544:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 9 81 theme glycoproteins 1798:1810 arg1 features 1776:1783 glycosylation features 1762:1783 glycosylation features of mammalian glycoproteins 1762:1810 CONCLUSIONS: Updates and improvements to UniCarbKB have introduced unique features for storing and displaying glycosylation features of mammalian glycoproteins. 26940363 7 82 theme glycoproteomics 1355:1369 arg1 data 1371:1374 large-scale glycoproteomics data 1343:1374 large-scale glycoproteomics data 1343:1374 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 0 83 theme compositional 77:89 arg1 data 107:110 compositional glycoproteomics data 77:110 compositional glycoproteomics data 77:110 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 10 84 gly glycoproteins 1974:1986 arg1 glycoproteins 1974:1986 glycoproteins 1974:1986 glycoproteins 1974:1986 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 2 85 theme global-specific 471:485 arg1 data 487:490 global-specific data 471:490 annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins 456:572 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 14 86 theme Special 2407:2413 arg1 Issue 2415:2419 a Special Issue 2405:2419 a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc 2405:2499 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 5 87 theme oligosaccharides 971:986 arg1 changes 960:966 structural and abundance changes 935:966 changes 960:966 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 8 88 theme efficient 1526:1534 arg1 representation 1536:1549 the efficient representation 1522:1549 the efficient representation of disease terms and associated changes in glycan heterogeneity 1522:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 4 89 from launch 790:795 arg1 2012 813:816 2012 813:816 2012 813:816 METHODS: Since the launch of UniCarbKB in 2012, we have continued to improve the organisation of our data model. 26940363 5 90 theme different 991:999 arg1 states 1015:1020 different human disease states 991:1020 different human disease states 991:1020 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 4 91 theme UniCarbKB 800:808 arg1 launch 790:795 the launch 786:795 the launch of UniCarbKB in 2012 786:816 METHODS: Since the launch of UniCarbKB in 2012, we have continued to improve the organisation of our data model. 26940363 8 92 theme terms 1562:1566 arg1 representation 1536:1549 the efficient representation 1522:1549 the efficient representation of disease terms and associated changes in glycan heterogeneity 1522:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 0 93 theme database 15:22 arg1 features 24:31 New database features 11:31 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 0:137 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 6 94 theme colorectal 1238:1247 arg1 cancer 1262:1267 prostate cancer 1253:1267 prostate cancer 1253:1267 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 94 theme colorectal 1238:1247 arg1 examples 1283:1290 examples 1283:1290 examples 1283:1290 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 94 theme colorectal 1238:1247 arg1 set 1221:1223 a set 1219:1223 a set of published colorectal and prostate cancer cell lines 1219:1278 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 6 94 theme colorectal 1238:1247 arg1 colorectal 1238:1247 colorectal 1238:1247 colorectal 1238:1247 RESULTS: In this manuscript, we demonstrate the capability of UniCarbKB to store and query relative glycan abundance data using a set of published colorectal and prostate cancer cell lines as examples. 26940363 13 95 theme wide 2330:2333 arg1 range 2335:2339 a wide range 2328:2339 a wide range of data with corresponding annotations 2328:2378 UniCarbKB was established with the mission of providing a resource for human glycobiology by capturing a wide range of data with corresponding annotations. 26940363 8 96 theme associated 1572:1581 arg1 changes 1583:1589 associated changes 1572:1589 associated changes in glycan heterogeneity 1572:1613 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 0 97 dep UniCarbKB 0:8 arg1 features 24:31 New database features 11:31 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 0:137 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 1 98 theme data 310:313 arg1 database 270:277 a curated database 260:277 a curated database 260:277 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 10 99 theme line 1932:1935 arg1 introduction 1911:1922 introduction 1911:1922 introduction of cell line 1911:1935 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 10 99 theme line 1932:1935 arg1 glycoproteomics 1891:1905 large-scale glycoproteomics 1879:1905 large-scale glycoproteomics 1879:1905 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 11 100 theme GENERAL 2009:2015 arg1 SIGNIFICANCE 2017:2028 GENERAL SIGNIFICANCE 2009:2028 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 2009:2140 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 0 101 theme glycan 49:54 arg1 abundance 66:74 glycan structure abundance 49:74 glycan structure abundance 49:74 UniCarbKB: New database features for integrating glycan structure abundance, compositional glycoproteomics data, and disease associations. 26940363 14 102 from Glycans 2431:2437 arg1 medicine 2455:2462 personalised medicine 2442:2462 personalised medicine" Guest Editor 2442:2476 This article is part of a Special Issue entitled "Glycans in personalised medicine" Guest Editor: Professor Gordan Lauc. 26940363 2 103 theme annotation 456:465 arg1 levels 446:451 two levels 442:451 two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity 442:642 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 10 104 theme glycomes 1999:2006 arg1 analysis 1962:1969 the analysis 1958:1969 the analysis of glycoproteins and entire glycomes 1958:2006 The integration of site-specific glycosylation data obtained from large-scale glycoproteomics and introduction of cell line studies will improve the analysis of glycoproteins and entire glycomes. 26940363 5 105 from changes 960:966 arg1 states 1015:1020 different human disease states 991:1020 different human disease states 991:1020 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 5 105 from changes 960:966 arg1 models 1039:1044 experimental models 1026:1044 experimental models 1026:1044 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 7 106 dep outline 1309:1315 arg1 value 1457:1461 adding value 1450:1461 adding value to UniCarbKB 1450:1474 Furthermore, we outline our strategy for managing large-scale glycoproteomics data, site-specific and glycan compositional data, and how this information is adding value to UniCarbKB. 26940363 2 107 dep oligosaccharides 495:510 arg1 characterised 525:537 characterised 525:537 characterised 525:537 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 2 107 dep oligosaccharides 495:510 arg1 released 512:519 released 512:519 released 512:519 The database comprises two levels of annotation (i) global-specific data of oligosaccharides released and characterised from single purified glycoproteins and (ii) information pertaining to site-specific glycan heterogeneity. 26940363 5 108 theme disease 1007:1013 arg1 states 1015:1020 different human disease states 991:1020 different human disease states 991:1020 Recently, we have extended our pipeline to collate structural and abundance changes of oligosaccharides in different human disease states and experimental models to extend our coverage of the human glycome. 26940363 11 109 theme data 2090:2093 arg1 types 2095:2099 new data types 2086:2099 new data types 2086:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 1 110 theme knowledge 237:245 arg1 representation 196:209 the representation 192:209 the representation of mammalian glycobiology knowledge 192:245 BACKGROUND: UniCarbKB aims to provide a resource for the representation of mammalian glycobiology knowledge by providing a curated database of structural and experimental data, supported by a web application that allows users to easily find and view richly annotated information. 26940363 11 111 dep SIGNIFICANCE 2017:2028 arg1 advancements 2042:2053 Continuing advancements 2031:2053 Continuing advancements in analytical technologies and new data types 2031:2099 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 11 111 dep SIGNIFICANCE 2017:2028 arg1 glycomics 2131:2139 advancing disease-related glycomics 2105:2139 advancing disease-related glycomics 2105:2139 GENERAL SIGNIFICANCE: Continuing advancements in analytical technologies and new data types are advancing disease-related glycomics. 26940363 8 112 theme Disease 1634:1640 arg1 Ontology 1642:1649 the Disease Ontology 1630:1649 the Disease Ontology 1630:1649 Finally, we summarise our efforts to improve the efficient representation of disease terms and associated changes in glycan heterogeneity by integrating the Disease Ontology. 26940363 3 113 theme taxonomic 733:741 arg1 information 743:753 structural, bibliographic, and taxonomic information 702:753 structural, bibliographic, and taxonomic information for each entry 702:768 Additional, contextual information is provided including structural, bibliographic, and taxonomic information for each entry. 26729457 10 0 from development 1967:1977 arg1 range 1878:1882 a range 1876:1882 a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control 1876:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 10 1 theme Q-TOF 1801:1805 arg1 platform 1835:1842 The Q-TOF based glycopeptide analysis platform 1797:1842 The Q-TOF based glycopeptide analysis platform presented here 1797:1857 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 8 2 gly O-glycopeptides 1476:1490 arg2 O-glycopeptides 1476:1490 O-glycopeptides 1476:1490 O-glycopeptides 1476:1490 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 9 3 theme LC-ESI 1748:1753 arg1 method 1765:1770 this optimized LC-ESI tandem MS method 1733:1770 this optimized LC-ESI tandem MS method with minimal user input 1733:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 10 4 theme based 1807:1811 arg1 platform 1835:1842 The Q-TOF based glycopeptide analysis platform 1797:1842 The Q-TOF based glycopeptide analysis platform presented here 1797:1857 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 9 5 theme MS 1762:1763 arg1 method 1765:1770 this optimized LC-ESI tandem MS method 1733:1770 this optimized LC-ESI tandem MS method with minimal user input 1733:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 2 6 gly O-glycopeptide 311:324 arg2 O-glycopeptide 311:324 O-glycopeptide 311:324 O-glycopeptide 311:324 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 4 7 gly disialylated 657:668 arg1 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 3 8 theme fragmentation 567:579 arg1 regimes 581:587 glycopeptide fragmentation regimes 554:587 glycopeptide fragmentation regimes 554:587 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 1 9 theme glycosylation 192:204 arg1 investigations 166:179 In-depth site-specific investigations 143:179 In-depth site-specific investigations of protein glycosylation 143:204 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 1 9 theme glycosylation 192:204 arg1 basis 214:218 the basis 210:218 the basis for understanding the biological function of glycoproteins 210:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 6 10 theme fragmentation 1035:1047 arg1 parameters 1049:1058 these optimized fragmentation parameters 1019:1058 these optimized fragmentation parameters 1019:1058 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 9 11 theme user 1785:1788 arg1 input 1790:1794 minimal user input 1777:1794 minimal user input 1777:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 4 12 theme disialylated 657:668 arg1 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 9 13 contain carrying 1675:1682 arg2 glycoforms 1699:1708 >110 different glycoforms 1684:1708 >110 different glycoforms 1684:1708 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 9 13 contain carrying 1675:1682 arg1 N-glycopeptides 1622:1636 Twenty-six out of 30 N-glycopeptides 1601:1636 Twenty-six out of 30 N-glycopeptides 1601:1636 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 3 14 theme data 603:606 arg1 analysis 608:615 automated data analysis 593:615 automated data analysis 593:615 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 5 15 gly glycopeptide 982:993 arg2 glycopeptide 982:993 glycopeptide fragmentation 982:1007 glycopeptide fragmentation 982:1007 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 5 16 theme optimal 874:880 arg1 energy 896:901 optimal fragmentation energy 874:901 optimal fragmentation energy 874:901 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 4 17 theme energy 766:771 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 17 theme energy 766:771 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 18 used used 627:630 arg2 we 624:625 we 624:625 we 624:625 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 8 19 theme glycan 1538:1543 arg1 compositions 1545:1556 the most abundant glycan compositions 1520:1556 the most abundant glycan compositions 1520:1556 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 4 20 theme collision 782:790 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 20 theme collision 782:790 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 2 21 dep occupancy 388:396 arg1 site 383:386 the glycosylation site, site occupancy 359:396 site 383:386 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 22 theme spectrometry-based 285:302 arg1 N- 304:305 Mass spectrometry-based N- 280:305 Mass spectrometry-based N- 280:305 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 5 23 theme glycopeptide 982:993 arg1 fragmentation 995:1007 glycopeptide fragmentation 982:1007 glycopeptide fragmentation 982:1007 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 6 24 theme higher-energy 1091:1103 arg1 CID 1105:1107 higher-energy CID 1091:1107 higher-energy CID 1091:1107 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 4 25 theme instrument 728:737 arg1 parameters 739:748 Q-TOF instrument parameters 722:748 Q-TOF instrument parameters 722:748 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 10 26 gly glycopeptide 1813:1824 arg2 glycopeptide 1813:1824 The Q-TOF based glycopeptide analysis platform 1797:1842 The Q-TOF based glycopeptide analysis platform presented here 1797:1857 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 1 27 theme site-specific 152:164 arg1 investigations 166:179 In-depth site-specific investigations 143:179 In-depth site-specific investigations of protein glycosylation 143:204 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 1 27 theme site-specific 152:164 arg1 basis 214:218 the basis 210:218 the basis for understanding the biological function of glycoproteins 210:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 8 28 theme putative 1580:1587 arg1 structures 1589:1598 putative structures 1580:1598 putative structures 1580:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 7 29 theme immunoglobulins 1285:1299 arg1 fetuin 1357:1362 bovine fetuin 1350:1362 a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin 1270:1362 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 29 theme immunoglobulins 1285:1299 arg1 set 1272:1274 a set 1270:1274 a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin 1270:1362 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 29 theme immunoglobulins 1285:1299 arg1 immunoglobulins 1285:1299 human immunoglobulins 1279:1299 human immunoglobulins 1279:1299 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 30 theme human 1279:1283 arg1 immunoglobulins 1285:1299 human immunoglobulins 1279:1299 human immunoglobulins 1279:1299 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 1 31 gly glycoproteins 265:277 arg1 glycoproteins 265:277 glycoproteins 265:277 glycoproteins 265:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 10 32 from applications 1897:1908 arg1 development 1967:1977 biopharmaceutical development 1949:1977 biopharmaceutical development 1949:1977 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 10 32 from applications 1897:1908 arg1 research 1929:1936 glycoproteomics research 1913:1936 glycoproteomics research as well as biopharmaceutical development and quality control 1913:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 7 33 theme bovine 1350:1355 arg1 fetuin 1357:1362 bovine fetuin 1350:1362 a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin 1270:1362 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 8 34 theme glycopeptide 1437:1448 arg1 assignment 1450:1459 software-assisted glycopeptide assignment 1419:1459 software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures 1419:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 5 35 theme fragmentation 882:894 arg1 energy 896:901 optimal fragmentation energy 874:901 optimal fragmentation energy 874:901 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 3 36 theme glycopeptide 554:565 arg1 regimes 581:587 glycopeptide fragmentation regimes 554:587 glycopeptide fragmentation regimes 554:587 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 10 37 theme biopharmaceutical 1949:1965 arg1 development 1967:1977 biopharmaceutical development 1949:1977 biopharmaceutical development 1949:1977 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 2 38 theme particular 440:449 arg1 site 451:454 a particular site 438:454 a particular site 438:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 5 39 theme linear 839:844 arg1 dependency 846:855 A linear dependency 837:855 A linear dependency of m/z-ratio and optimal fragmentation energy 837:901 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 8 40 theme N- 1469:1470 arg1 assignment 1450:1459 software-assisted glycopeptide assignment 1419:1459 software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures 1419:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 2 41 theme occupancy 388:396 arg1 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 41 theme occupancy 388:396 arg1 varieties 417:425 glycan varieties 410:425 glycan varieties present on a particular site 410:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 41 theme occupancy 388:396 arg1 determination 342:354 determination 342:354 determination of the glycosylation site, site occupancy 342:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 0 42 theme Collision 35:43 arg1 Energies 45:52 Optimizing Collision Energies 24:52 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 0:141 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 2 43 attach present 427:433 arg2 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 43 attach present 427:433 arg1 site 451:454 a particular site 438:454 a particular site 438:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 43 attach present 427:433 arg2 varieties 417:425 glycan varieties 410:425 glycan varieties present on a particular site 410:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 6 44 theme single 1206:1211 arg1 experiment 1223:1232 a single tandem MS experiment 1204:1232 a single tandem MS experiment 1204:1232 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 8 45 theme O-glycopeptides 1476:1490 arg1 assignment 1450:1459 software-assisted glycopeptide assignment 1419:1459 software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures 1419:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 1 46 theme biological 242:251 arg1 function 253:260 the biological function 238:260 the biological function of glycoproteins 238:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 10 47 theme analysis 1826:1833 arg1 platform 1835:1842 The Q-TOF based glycopeptide analysis platform 1797:1842 The Q-TOF based glycopeptide analysis platform presented here 1797:1857 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 6 48 theme peptide-specific 1170:1185 arg1 fragments 1187:1195 glycan and peptide-specific fragments 1159:1195 fragments 1187:1195 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 4 49 gly glycopeptides 822:834 arg2 glycopeptides 822:834 glycopeptides 822:834 glycopeptides 822:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 50 gly N-glycopeptides 683:697 arg2 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 3 51 theme analytical 526:535 arg1 regimes 581:587 glycopeptide fragmentation regimes 554:587 glycopeptide fragmentation regimes 554:587 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 3 51 theme analytical 526:535 arg1 tools 537:541 the applied analytical tools 514:541 the applied analytical tools 514:541 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 3 51 theme analytical 526:535 arg1 analysis 608:615 automated data analysis 593:615 automated data analysis 593:615 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 9 52 theme optimized 1738:1746 arg1 method 1765:1770 this optimized LC-ESI tandem MS method 1733:1770 this optimized LC-ESI tandem MS method with minimal user input 1733:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 6 53 theme glycan 1159:1164 arg1 fragments 1187:1195 glycan and peptide-specific fragments 1159:1195 fragments 1187:1195 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 9 54 theme different 1689:1697 arg1 glycoforms 1699:1708 >110 different glycoforms 1684:1708 >110 different glycoforms 1684:1708 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 9 55 theme tandem 1755:1760 arg1 method 1765:1770 this optimized LC-ESI tandem MS method 1733:1770 this optimized LC-ESI tandem MS method with minimal user input 1733:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 2 56 theme O-glycopeptide 311:324 arg1 analyses 326:333 Mass spectrometry-based N- and O-glycopeptide analyses 280:333 Mass spectrometry-based N- and O-glycopeptide analyses 280:333 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 1 57 theme protein 184:190 arg1 glycosylation 192:204 protein glycosylation 184:204 protein glycosylation 184:204 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 4 58 theme N-glycopeptides 683:697 arg1 set 640:642 a small set 632:642 a small set of synthetic disialylated, biantennary N-glycopeptides 632:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 58 theme N-glycopeptides 683:697 arg1 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 2 59 theme glycosylation 363:375 arg1 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 9 60 theme minimal 1777:1783 arg1 input 1790:1794 minimal user input 1777:1794 minimal user input 1777:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 0 61 theme Glycopeptide-Based 107:124 arg1 Glycoproteomics 126:140 Glycopeptide-Based Glycoproteomics 107:140 Glycopeptide-Based Glycoproteomics 107:140 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 3 62 theme automated 593:601 arg1 analysis 608:615 automated data analysis 593:615 automated data analysis 593:615 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 5 63 theme energy 896:901 arg1 dependency 846:855 A linear dependency 837:855 A linear dependency of m/z-ratio and optimal fragmentation energy 837:901 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 6 64 theme optimized 1025:1033 arg1 parameters 1049:1058 these optimized fragmentation parameters 1019:1058 these optimized fragmentation parameters 1019:1058 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 6 65 theme MS 1220:1221 arg1 experiment 1223:1232 a single tandem MS experiment 1204:1232 a single tandem MS experiment 1204:1232 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 4 66 theme synthetic 647:655 arg1 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 10 67 theme applications 1897:1908 arg1 range 1878:1882 a range 1876:1882 a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control 1876:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 8 68 theme abundant 1529:1536 arg1 compositions 1545:1556 the most abundant glycan compositions 1520:1556 the most abundant glycan compositions 1520:1556 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 4 69 theme optimal 758:764 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 69 theme optimal 758:764 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 2 70 theme N- 304:305 arg1 analyses 326:333 Mass spectrometry-based N- and O-glycopeptide analyses 280:333 Mass spectrometry-based N- and O-glycopeptide analyses 280:333 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 8 71 theme peptide 1559:1565 arg1 sequence 1567:1574 peptide sequence 1559:1574 peptide sequence 1559:1574 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 8 72 theme optimized 1371:1379 arg1 parameters 1395:1404 These optimized fragmentation parameters 1365:1404 These optimized fragmentation parameters 1365:1404 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 4 73 theme stepping 773:780 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 73 theme stepping 773:780 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 10 74 theme glycoproteomics 1913:1927 arg1 research 1929:1936 glycoproteomics research 1913:1936 glycoproteomics research as well as biopharmaceutical development and quality control 1913:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 1 75 theme In-depth 143:150 arg1 investigations 166:179 In-depth site-specific investigations 143:179 In-depth site-specific investigations of protein glycosylation 143:204 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 1 75 theme In-depth 143:150 arg1 basis 214:218 the basis 210:218 the basis for understanding the biological function of glycoproteins 210:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 26729457 4 76 theme Q-TOF 722:726 arg1 parameters 739:748 Q-TOF instrument parameters 722:748 Q-TOF instrument parameters 722:748 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 0 77 theme Flight 76:81 arg1 Instruments 91:101 Flight (Q-TOF) Instruments 76:101 Flight (Q-TOF) Instruments 76:101 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 3 78 gly glycopeptide 554:565 arg2 glycopeptide 554:565 glycopeptide fragmentation regimes 554:587 glycopeptide fragmentation regimes 554:587 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 0 79 theme Q-TOF 84:88 arg1 Instruments 91:101 Flight (Q-TOF) Instruments 76:101 Flight (Q-TOF) Instruments 76:101 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 0 80 from Energies 45:52 arg1 Quadrupole-Time 57:71 Quadrupole-Time 57:71 Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics 57:140 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 9 81 with method 1765:1770 arg1 input 1790:1794 minimal user input 1777:1794 minimal user input 1777:1794 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 0 82 theme Instruments 91:101 arg1 Quadrupole-Time 57:71 Quadrupole-Time 57:71 Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics 57:140 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 10 83 from range 1878:1882 arg1 development 1967:1977 biopharmaceutical development 1949:1977 biopharmaceutical development 1949:1977 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 10 83 from range 1878:1882 arg1 research 1929:1936 glycoproteomics research 1913:1936 glycoproteomics research as well as biopharmaceutical development and quality control 1913:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 8 84 theme fragmentation 1381:1393 arg1 parameters 1395:1404 These optimized fragmentation parameters 1365:1404 These optimized fragmentation parameters 1365:1404 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 2 85 theme present 427:433 arg1 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 85 theme present 427:433 arg1 varieties 417:425 glycan varieties 410:425 glycan varieties present on a particular site 410:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 5 86 theme m/z-ratio 860:868 arg1 dependency 846:855 A linear dependency 837:855 A linear dependency of m/z-ratio and optimal fragmentation energy 837:901 A linear dependency of m/z-ratio and optimal fragmentation energy was found, showing that with increasing m/z-ratio, more energy is required for glycopeptide fragmentation. 26729457 6 87 theme online 1137:1142 arg1 acquisition 1144:1154 the online acquisition 1133:1154 the online acquisition of glycan and peptide-specific fragments 1133:1195 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 9 88 gly N-glycopeptides 1622:1636 arg2 N-glycopeptides 1622:1636 Twenty-six out of 30 N-glycopeptides 1601:1636 Twenty-six out of 30 N-glycopeptides 1601:1636 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 4 89 theme induced 792:798 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 89 theme induced 792:798 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 2 90 theme glycan 410:415 arg1 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 90 theme glycan 410:415 arg1 varieties 417:425 glycan varieties 410:425 glycan varieties present on a particular site 410:454 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 0 91 theme Destruction 11:21 arg1 Art 4:6 The Art 0:6 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 0:141 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 8 92 theme software-assisted 1419:1435 arg1 assignment 1450:1459 software-assisted glycopeptide assignment 1419:1459 software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures 1419:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 0 93 theme Optimizing 24:33 arg1 Energies 45:52 Optimizing Collision Energies 24:52 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 0:141 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 10 94 from research 1929:1936 arg1 range 1878:1882 a range 1876:1882 a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control 1876:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 3 95 theme information 479:489 arg1 dependent 501:509 dependent 501:509 dependent 501:509 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 3 95 theme information 479:489 arg1 depth 470:474 the depth 466:474 the depth of information 466:489 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 4 96 dep disialylated 657:668 arg1 biantennary 671:681 biantennary 671:681 biantennary 671:681 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 6 97 theme tandem 1213:1218 arg1 experiment 1223:1232 a single tandem MS experiment 1204:1232 a single tandem MS experiment 1204:1232 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 9 98 gly O-glycopeptides 1659:1673 arg2 O-glycopeptides 1659:1673 four out of five O-glycopeptides 1642:1673 four out of five O-glycopeptides 1642:1673 Twenty-six out of 30 N-glycopeptides and four out of five O-glycopeptides carrying >110 different glycoforms could be identified by this optimized LC-ESI tandem MS method with minimal user input. 26729457 7 99 dep set 1272:1274 arg1 sIgA 1310:1313 sIgA 1310:1313 sIgA 1310:1313 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 99 dep set 1272:1274 arg1 IgG1+2 1316:1321 IgG1+2 1316:1321 IgG1+2 1316:1321 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 99 dep set 1272:1274 arg1 IgM 1334:1336 IgM 1334:1336 IgM 1334:1336 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 99 dep set 1272:1274 arg1 IgA1+2 1302:1307 IgA1+2 1302:1307 IgA1+2 1302:1307 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 99 dep set 1272:1274 arg1 IgE 1324:1326 IgE 1324:1326 IgE 1324:1326 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 7 99 dep set 1272:1274 arg1 IgD 1329:1331 IgD 1329:1331 IgD 1329:1331 We validated this method analyzing a set of human immunoglobulins (IgA1+2, sIgA, IgG1+2, IgE, IgD, IgM) as well as bovine fetuin. 26729457 3 100 theme applied 518:524 arg1 regimes 581:587 glycopeptide fragmentation regimes 554:587 glycopeptide fragmentation regimes 554:587 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 3 100 theme applied 518:524 arg1 tools 537:541 the applied analytical tools 514:541 the applied analytical tools 514:541 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 3 100 theme applied 518:524 arg1 analysis 608:615 automated data analysis 593:615 automated data analysis 593:615 However, the depth of information is highly dependent on the applied analytical tools, including glycopeptide fragmentation regimes and automated data analysis. 26729457 8 101 gly glycopeptide 1437:1448 arg2 glycopeptide 1437:1448 software-assisted glycopeptide assignment 1419:1459 software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures 1419:1598 These optimized fragmentation parameters also enabled software-assisted glycopeptide assignment of both N- and O-glycopeptides including information about the most abundant glycan compositions, peptide sequence and putative structures. 26729457 10 102 theme different 1887:1895 arg1 applications 1897:1908 different applications 1887:1908 different applications in glycoproteomics research as well as biopharmaceutical development and quality control 1887:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 4 103 theme small 634:638 arg1 set 640:642 a small set 632:642 a small set of synthetic disialylated, biantennary N-glycopeptides 632:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 103 theme small 634:638 arg1 N-glycopeptides 683:697 synthetic disialylated, biantennary N-glycopeptides 647:697 synthetic disialylated, biantennary N-glycopeptides 647:697 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 104 theme glycopeptides 822:834 arg1 CID 814:816 CID 814:816 CID 814:816 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 4 104 theme glycopeptides 822:834 arg1 dissociation 800:811 optimal energy stepping collision induced dissociation 758:811 optimal energy stepping collision induced dissociation (CID) of glycopeptides 758:834 Here, we used a small set of synthetic disialylated, biantennary N-glycopeptides to systematically tune Q-TOF instrument parameters towards optimal energy stepping collision induced dissociation (CID) of glycopeptides. 26729457 10 105 theme quality 1983:1989 arg1 control 1991:1997 quality control 1983:1997 quality control 1983:1997 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 2 106 theme site 377:380 arg1 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 0 107 dep Art 4:6 arg1 Energies 45:52 Optimizing Collision Energies 24:52 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 0:141 The Art of Destruction: Optimizing Collision Energies in Quadrupole-Time of Flight (Q-TOF) Instruments for Glycopeptide-Based Glycoproteomics. 26729457 6 108 theme fragments 1187:1195 arg1 acquisition 1144:1154 the online acquisition 1133:1154 the online acquisition of glycan and peptide-specific fragments 1133:1195 Based on these optimized fragmentation parameters, a method combining lower- and higher-energy CID was developed, allowing the online acquisition of glycan and peptide-specific fragments within a single tandem MS experiment. 26729457 2 109 gly glycosylation 363:375 arg2 site 377:380 the glycosylation site, site occupancy 359:396 site 377:380 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 2 109 gly glycosylation 363:375 arg2 occupancy 388:396 the glycosylation site, site occupancy 359:396 occupancy 388:396 Mass spectrometry-based N- and O-glycopeptide analyses enable determination of the glycosylation site, site occupancy, as well as glycan varieties present on a particular site. 26729457 10 110 theme glycopeptide 1813:1824 arg1 platform 1835:1842 The Q-TOF based glycopeptide analysis platform 1797:1842 The Q-TOF based glycopeptide analysis platform presented here 1797:1857 The Q-TOF based glycopeptide analysis platform presented here opens the way to a range of different applications in glycoproteomics research as well as biopharmaceutical development and quality control. 26729457 1 111 theme glycoproteins 265:277 arg1 function 253:260 the biological function 238:260 the biological function of glycoproteins 238:277 In-depth site-specific investigations of protein glycosylation are the basis for understanding the biological function of glycoproteins. 27573070 9 0 theme tandem 1766:1771 arg1 spectra 1778:1784 the tandem mass spectra 1762:1784 the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans 1762:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 3 1 theme simultaneous 697:708 arg1 integration 710:720 the simultaneous integration 693:720 the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation 693:821 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 1 2 located observed 194:201 arg1 diseases 214:221 various diseases 206:221 various diseases 206:221 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 1 2 located observed 194:201 arg2 changes 151:157 changes 151:157 changes in protein glycosylation 151:182 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 10 3 from http 1927:1930 arg1 available 1905:1913 available 1905:1913 available 1905:1913 gFinder is available publicly at http://gFinder.proteomix.org/ . 27573070 6 4 theme convenient 1227:1236 arg1 functions 1238:1246 many convenient functions 1222:1246 many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen 1222:1332 gFinder also provides many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen. 27573070 6 5 theme spectrum 1315:1322 arg1 on-screen 1324:1332 the spectrum on-screen 1311:1332 the spectrum on-screen 1311:1332 gFinder also provides many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen. 27573070 3 6 theme web-based 551:559 arg1 tool 576:579 a web-based bioinformatics tool 549:579 a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis 549:878 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 6 theme web-based 551:559 arg1 gFinder 540:546 gFinder 540:546 gFinder 540:546 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 4 7 from data 988:991 arg1 identification 915:928 the identification 911:928 the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 911:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 9 8 theme convenient 1701:1710 arg1 tool 1740:1743 a convenient, high-throughput analytical tool 1699:1743 a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans 1699:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 9 8 theme convenient 1701:1710 arg1 gFinder 1688:1694 gFinder 1688:1694 gFinder 1688:1694 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 7 9 theme N-linked 1458:1465 arg1 glycosylation 1467:1479 N-linked glycosylation 1458:1479 N-linked glycosylation 1458:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 3 10 theme dissociation 743:754 arg1 integration 710:720 the simultaneous integration 693:720 the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation 693:821 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 11 theme collisional 778:788 arg1 HCD 804:806 HCD 804:806 HCD 804:806 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 11 theme collisional 778:788 arg1 dissociation 790:801 high-energy collisional dissociation 766:801 high-energy collisional dissociation (HCD) 766:807 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 7 12 theme published 1428:1436 arg1 set 1443:1445 the previously published data set 1413:1445 the previously published data set containing N-linked glycosylation 1413:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 3 13 gly N-glycopeptides 614:628 arg2 N-glycopeptides 614:628 native N-glycopeptides 607:628 native N-glycopeptides 607:628 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 4 14 theme MS 985:986 arg1 data 988:991 tandem MS data 978:991 tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 978:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 8 15 theme N-glycan 1486:1493 arg1 analysis 1495:1502 N-glycan analysis 1486:1502 N-glycan analysis 1486:1502 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 6 16 theme manual 1277:1282 arg1 calculations 1284:1295 manual calculations 1277:1295 manual calculations 1277:1295 gFinder also provides many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen. 27573070 3 17 theme N-glycopeptides 614:628 arg1 mixtures 595:602 mixtures 595:602 mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis 595:878 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 4 18 theme glycans 938:944 arg1 identification 915:928 the identification 911:928 the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 911:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 8 19 used used 1508:1511 arg2 we 1505:1506 we 1505:1506 we 1505:1506 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 4 20 theme N-glycopeptide 950:963 arg1 backbones 965:973 N-glycopeptide backbones 950:973 N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 950:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 4 21 from glycans 938:944 arg1 data 988:991 tandem MS data 978:991 tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 978:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 2 22 theme glycosylation 284:296 arg1 sites 298:302 glycosylation sites 284:302 glycosylation sites 284:302 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 9 23 used used 1824:1827 arg2 N-glycopeptides 1789:1803 N-glycopeptides 1789:1803 N-glycopeptides 1789:1803 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 9 24 theme N-glycopeptides 1789:1803 arg1 spectra 1778:1784 the tandem mass spectra 1762:1784 the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans 1762:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 8 25 theme GlycomeDB 1517:1525 arg1 database 1544:1551 the GlycomeDB glycan structure database 1513:1551 the GlycomeDB glycan structure database 1513:1551 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 5 26 theme possible 1148:1155 arg1 structures 1166:1175 possible N-glycan structures 1148:1175 possible N-glycan structures 1148:1175 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 2 27 theme glycoprotein 267:278 arg1 characterization 247:262 characterization 247:262 characterization 247:262 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 27 theme glycoprotein 267:278 arg1 identification 228:241 identification 228:241 identification 228:241 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 4 28 theme proteomic 1025:1033 arg1 Mascot 1054:1059 Mascot 1054:1059 Mascot 1054:1059 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 4 28 theme proteomic 1025:1033 arg1 tool 1042:1045 a proteomic search tool 1023:1045 a proteomic search tool (e.g., Mascot) 1023:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 8 29 theme structure 1534:1542 arg1 database 1544:1551 the GlycomeDB glycan structure database 1513:1551 the GlycomeDB glycan structure database 1513:1551 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 3 30 theme high-throughput 855:869 arg1 analysis 871:878 high-throughput analysis 855:878 high-throughput analysis 855:878 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 0 31 theme N-Glycopeptides 61:75 arg1 Analysis 49:56 the Analysis 45:56 the Analysis of N-Glycopeptides 45:75 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 27573070 4 32 gly N-glycopeptide 950:963 arg2 N-glycopeptide 950:963 N-glycopeptide backbones 950:973 N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 950:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 7 33 link N-linked 1458:1465 arg1 glycosylation 1467:1479 N-linked glycosylation 1458:1479 N-linked glycosylation 1458:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 5 34 theme peptide 1117:1123 arg1 sequences 1134:1142 peptide backbone sequences 1117:1142 peptide backbone sequences 1117:1142 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 1 35 from changes 151:157 arg1 glycosylation 170:182 protein glycosylation 162:182 protein glycosylation 162:182 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 4 36 theme glycan 1003:1008 arg1 database 1010:1017 the glycan database 999:1017 the glycan database 999:1017 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 9 37 theme proteins 1869:1876 arg1 identification 1833:1846 identification 1833:1846 identification of potential missing proteins having glycans 1833:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 8 38 theme structural 1575:1584 arg1 data 1600:1603 the structural and taxonomic data 1571:1603 the structural and taxonomic data from all of the major carbohydrate databases available in the public domain 1571:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 7 39 theme additional 1364:1373 arg1 Q8N9B8 1384:1389 Q8N9B8 1384:1389 Q8N9B8 1384:1389 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 7 39 theme additional 1364:1373 arg1 protein 1375:1381 an additional protein 1361:1381 an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation 1361:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 9 40 theme potential 1851:1859 arg1 proteins 1869:1876 potential missing proteins 1851:1876 potential missing proteins having glycans 1851:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 8 41 theme taxonomic 1590:1598 arg1 data 1600:1603 the structural and taxonomic data 1571:1603 the structural and taxonomic data from all of the major carbohydrate databases available in the public domain 1571:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 7 42 contain containing 1447:1456 arg1 set 1443:1445 the previously published data set 1413:1445 the previously published data set containing N-linked glycosylation 1413:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 7 42 contain containing 1447:1456 arg2 glycosylation 1467:1479 N-linked glycosylation 1458:1479 N-linked glycosylation 1458:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 3 43 theme gFinder 540:546 arg1 development 525:535 the development 521:535 the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis 521:878 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 1 44 theme indispensable 111:123 arg1 functions 136:144 numerous indispensable biological functions 102:144 numerous indispensable biological functions 102:144 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 2 45 theme glycopeptides 488:500 arg1 analysis 464:471 the MS analysis 457:471 the MS analysis of glycans and glycopeptides 457:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 46 theme informatics 423:433 arg1 tools 435:439 proteome informatics tools 414:439 proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides 414:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 0 47 theme Bioinformatics 21:34 arg1 Tool 36:39 A Web-Based Bioinformatics Tool 9:39 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 0:76 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 27573070 3 48 dep dissociation 743:754 arg1 fragmentation 809:821 fragmentation 809:821 fragmentation 809:821 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 2 49 gly glycopeptides 488:500 arg2 glycopeptides 488:500 glycopeptides 488:500 glycopeptides 488:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 50 theme glycans 476:482 arg1 analysis 464:471 the MS analysis 457:471 the MS analysis of glycans and glycopeptides 457:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 5 51 theme assigned 1183:1190 arg1 scores 1192:1197 assigned scores 1183:1197 assigned scores 1183:1197 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 2 52 theme great 360:364 arg1 efforts 366:372 great efforts 360:372 great efforts 360:372 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 8 53 theme major 1621:1625 arg1 databases 1640:1648 the major carbohydrate databases 1617:1648 the major carbohydrate databases available in the public domain 1617:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 0 54 dep gFinder 0:6 arg1 Tool 36:39 A Web-Based Bioinformatics Tool 9:39 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 0:76 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 27573070 3 55 theme collision-induced 725:741 arg1 CID 757:759 CID 757:759 CID 757:759 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 55 theme collision-induced 725:741 arg1 dissociation 743:754 collision-induced dissociation 725:754 collision-induced dissociation (CID) 725:760 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 8 56 from all 1610:1612 arg1 data 1600:1603 the structural and taxonomic data 1571:1603 the structural and taxonomic data from all of the major carbohydrate databases available in the public domain 1571:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 4 57 from identification 915:928 arg1 data 988:991 tandem MS data 978:991 tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 978:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 8 58 theme public 1667:1672 arg1 domain 1674:1679 the public domain 1663:1679 the public domain 1663:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 6 59 theme many 1222:1225 arg1 functions 1238:1246 many convenient functions 1222:1246 many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen 1222:1332 gFinder also provides many convenient functions that make it easy to perform manual calculations while viewing the spectrum on-screen. 27573070 2 60 theme mass 307:310 arg1 spectrometry 312:323 mass spectrometry 307:323 mass spectrometry (MS) 307:328 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 60 theme mass 307:310 arg1 MS 326:327 MS 326:327 MS 326:327 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 61 gly glycosylation 284:296 arg2 sites 298:302 glycosylation sites 284:302 glycosylation sites 284:302 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 4 62 dep Mascot 1054:1059 arg1 e.g. 1048:1051 e.g. 1048:1051 e.g. 1048:1051 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 2 63 theme challenging 337:347 arg1 tasks 349:353 challenging tasks 337:353 challenging tasks 337:353 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 4 64 theme merged 887:892 arg1 spectra 894:900 These merged spectra 881:900 These merged spectra 881:900 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 2 65 gly glycoprotein 267:278 arg1 glycoprotein 267:278 glycoprotein 267:278 glycoprotein 267:278 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 3 66 theme dissociation 790:801 arg1 integration 710:720 the simultaneous integration 693:720 the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation 693:821 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 67 theme bioinformatics 561:574 arg1 tool 576:579 a web-based bioinformatics tool 549:579 a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis 549:878 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 67 theme bioinformatics 561:574 arg1 gFinder 540:546 gFinder 540:546 gFinder 540:546 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 1 68 theme various 206:212 arg1 diseases 214:221 various diseases 206:221 various diseases 206:221 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 4 69 theme backbones 965:973 arg1 identification 915:928 the identification 911:928 the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 911:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 7 70 theme data 1438:1441 arg1 set 1443:1445 the previously published data set 1413:1445 the previously published data set containing N-linked glycosylation 1413:1479 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 2 71 dep identification 228:241 arg1 The 224:226 The 224:226 The 224:226 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 3 72 theme high-energy 766:776 arg1 HCD 804:806 HCD 804:806 HCD 804:806 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 3 72 theme high-energy 766:776 arg1 dissociation 790:801 high-energy collisional dissociation 766:801 high-energy collisional dissociation (HCD) 766:807 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 8 73 from available 1650:1658 arg1 domain 1674:1679 the public domain 1663:1679 the public domain 1663:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 9 74 theme analytical 1729:1738 arg1 tool 1740:1743 a convenient, high-throughput analytical tool 1699:1743 a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans 1699:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 9 74 theme analytical 1729:1738 arg1 gFinder 1688:1694 gFinder 1688:1694 gFinder 1688:1694 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 3 75 theme native 607:612 arg1 N-glycopeptides 614:628 native N-glycopeptides 607:628 native N-glycopeptides 607:628 Here we report on the development of gFinder, a web-based bioinformatics tool that analyzes mixtures of native N-glycopeptides that have been profiled by tandem MS. gFinder not only enables the simultaneous integration of collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) fragmentation but also merges the spectra for high-throughput analysis. 27573070 4 76 theme tandem 978:983 arg1 data 988:991 tandem MS data 978:991 tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 978:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 4 77 from backbones 965:973 arg1 data 988:991 tandem MS data 978:991 tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot) 978:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 9 78 gly N-glycopeptides 1789:1803 arg2 N-glycopeptides 1789:1803 N-glycopeptides 1789:1803 N-glycopeptides 1789:1803 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 1 79 theme biological 125:134 arg1 functions 136:144 numerous indispensable biological functions 102:144 numerous indispensable biological functions 102:144 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 9 80 theme mass 1773:1776 arg1 spectra 1778:1784 the tandem mass spectra 1762:1784 the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans 1762:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 5 81 used used 1081:1084 arg2 data 1069:1072 These data 1063:1072 These data 1063:1072 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 8 82 from domain 1674:1679 arg1 available 1650:1658 available 1650:1658 available 1650:1658 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 2 83 theme sites 298:302 arg1 characterization 247:262 characterization 247:262 characterization 247:262 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 2 83 theme sites 298:302 arg1 identification 228:241 identification 228:241 identification 228:241 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 8 84 theme glycan 1527:1532 arg1 database 1544:1551 the GlycomeDB glycan structure database 1513:1551 the GlycomeDB glycan structure database 1513:1551 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 9 85 contain having 1878:1883 arg2 glycans 1885:1891 glycans 1885:1891 glycans 1885:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 9 85 contain having 1878:1883 arg1 proteins 1869:1876 potential missing proteins 1851:1876 potential missing proteins having glycans 1851:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 5 86 theme N-glycan 1157:1164 arg1 structures 1166:1175 possible N-glycan structures 1148:1175 possible N-glycan structures 1148:1175 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 1 87 theme protein 162:168 arg1 glycosylation 170:182 protein glycosylation 162:182 protein glycosylation 162:182 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 5 88 theme backbone 1125:1132 arg1 sequences 1134:1142 peptide backbone sequences 1117:1142 peptide backbone sequences 1117:1142 These data can be used to simultaneously characterize peptide backbone sequences and possible N-glycan structures using assigned scores. 27573070 2 89 theme MS 461:462 arg1 analysis 464:471 the MS analysis 457:471 the MS analysis of glycans and glycopeptides 457:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 7 90 used used 1338:1341 arg2 We 1335:1336 We 1335:1336 We 1335:1336 We used gFinder to detect an additional protein (Q8N9B8) that was missed from the previously published data set containing N-linked glycosylation. 27573070 0 91 theme Web-Based 11:19 arg1 Tool 36:39 A Web-Based Bioinformatics Tool 9:39 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 0:76 gFinder: A Web-Based Bioinformatics Tool for the Analysis of N-Glycopeptides. 27573070 1 92 theme numerous 102:109 arg1 functions 136:144 numerous indispensable biological functions 102:144 numerous indispensable biological functions 102:144 Glycoproteins influence numerous indispensable biological functions, and changes in protein glycosylation have been observed in various diseases. 27573070 9 93 theme missing 1861:1867 arg1 proteins 1869:1876 potential missing proteins 1851:1876 potential missing proteins having glycans 1851:1891 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 2 94 theme tools 435:439 arg1 development 399:409 the development 395:409 the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides 395:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 4 95 theme search 1035:1040 arg1 Mascot 1054:1059 Mascot 1054:1059 Mascot 1054:1059 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 4 95 theme search 1035:1040 arg1 tool 1042:1045 a proteomic search tool 1023:1045 a proteomic search tool (e.g., Mascot) 1023:1060 These merged spectra expedite the identification of both glycans and N-glycopeptide backbones in tandem MS data using the glycan database and a proteomic search tool (e.g., Mascot). 27573070 2 96 theme proteome 414:421 arg1 tools 435:439 proteome informatics tools 414:439 proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides 414:500 The identification and characterization of glycoprotein and glycosylation sites by mass spectrometry (MS) remain challenging tasks, and great efforts have been devoted to the development of proteome informatics tools that facilitate the MS analysis of glycans and glycopeptides. 27573070 9 97 dep convenient 1701:1710 arg1 high-throughput 1713:1727 high-throughput 1713:1727 high-throughput 1713:1727 Thus, gFinder is a convenient, high-throughput analytical tool for interpreting the tandem mass spectra of N-glycopeptides, which can then be used for identification of potential missing proteins having glycans. 27573070 8 98 theme carbohydrate 1627:1638 arg1 databases 1640:1648 the major carbohydrate databases 1617:1648 the major carbohydrate databases available in the public domain 1617:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27573070 8 99 theme available 1650:1658 arg1 databases 1640:1648 the major carbohydrate databases 1617:1648 the major carbohydrate databases available in the public domain 1617:1679 For N-glycan analysis, we used the GlycomeDB glycan structure database, which integrates the structural and taxonomic data from all of the major carbohydrate databases available in the public domain. 27111718 9 0 theme MS/MS 1754:1758 arg1 analysis 1760:1767 MS/MS analysis 1754:1767 MS/MS analysis 1754:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 3 1 theme spectrometry 496:507 arg1 instrumentation 509:523 mass spectrometry instrumentation 491:523 mass spectrometry instrumentation 491:523 While mass spectrometry instrumentation has seen advancements in capabilities, discovering glycosylation-related markers using existing software is currently not straightforward. 27111718 9 2 used used 1681:1684 arg2 GlycoSeq 1657:1664 GlycoSeq 1657:1664 GlycoSeq 1657:1664 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 6 3 theme glycan 1281:1286 arg1 structures 1288:1297 improbable glycan structures 1270:1297 improbable glycan structures 1270:1297 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 3 4 theme mass 491:494 arg1 spectrometry 496:507 mass spectrometry 491:507 mass spectrometry instrumentation 491:523 While mass spectrometry instrumentation has seen advancements in capabilities, discovering glycosylation-related markers using existing software is currently not straightforward. 27111718 4 5 from identification 728:741 arg1 samples 770:776 samples 770:776 samples 770:776 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 1 6 theme blood 179:183 arg1 samples 185:191 human blood samples 173:191 human blood samples 173:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 2 7 theme modifications 306:318 arg1 glycosylation 347:359 protein glycosylation 339:359 protein glycosylation 339:359 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 7 theme modifications 306:318 arg1 PTMs 321:324 PTMs 321:324 PTMs 321:324 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 7 theme modifications 306:318 arg1 modifications 306:318 the most common protein post-translational modifications 263:318 the most common protein post-translational modifications (PTMs) in mammals 263:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 7 theme modifications 306:318 arg1 one 256:258 one 256:258 one 256:258 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 5 8 theme open-source 918:928 arg1 tool 939:942 an open-source software tool 915:942 an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 915:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 5 8 theme open-source 918:928 arg1 GlycoSeq 905:912 GlycoSeq 905:912 GlycoSeq 905:912 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 6 9 theme glycan 1231:1236 arg1 pathways 1248:1255 glycan synthetic pathways 1231:1255 glycan synthetic pathways 1231:1255 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 5 10 theme tandem 1150:1155 arg1 spectrum 1162:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 2 11 theme protein 279:285 arg1 PTMs 321:324 PTMs 321:324 PTMs 321:324 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 11 theme protein 279:285 arg1 modifications 306:318 the most common protein post-translational modifications 263:318 the most common protein post-translational modifications (PTMs) in mammals 263:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 1 12 theme experimental 119:130 arg1 tool 132:135 a routine experimental tool 109:135 a routine experimental tool for proteomic biomarker analysis of human blood samples 109:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 4 13 theme protein 693:699 arg1 glycosylation 701:713 protein glycosylation 693:713 protein glycosylation 693:713 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 3 14 theme existing 612:619 arg1 software 621:628 existing software 612:628 existing software 612:628 While mass spectrometry instrumentation has seen advancements in capabilities, discovering glycosylation-related markers using existing software is currently not straightforward. 27111718 5 15 theme heuristic 962:970 arg1 algorithm 999:1007 a heuristic iterated glycan sequencing algorithm 960:1007 a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 960:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 9 16 from analysis 1760:1767 arg1 glycopeptides 1735:1747 glycopeptides 1735:1747 glycopeptides from MS/MS analysis 1735:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 9 16 from analysis 1760:1767 arg1 structures 1721:1730 glycan structures 1714:1730 glycan structures in glycopeptides from MS/MS analysis 1714:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 7 17 theme cancer 1446:1451 arg1 patients 1453:1460 breast cancer patients 1439:1460 breast cancer patients 1439:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 6 18 theme reasonable 1309:1318 arg1 trees 1327:1331 build reasonable glycan trees 1303:1331 build reasonable glycan trees 1303:1331 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 8 19 theme enzymatic 1479:1487 arg1 specificity 1489:1499 enzymatic specificity 1479:1499 enzymatic specificity 1479:1499 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 5 20 theme collision-induced 1119:1135 arg1 dissociation 1137:1148 its collision-induced dissociation 1115:1148 its collision-induced dissociation 1115:1148 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 5 21 theme glycan 981:986 arg1 algorithm 999:1007 a heuristic iterated glycan sequencing algorithm 960:1007 a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 960:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 4 22 theme intact 746:751 arg1 glycopeptides 753:765 intact glycopeptides 746:765 intact glycopeptides 746:765 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 4 22 theme intact 746:751 arg1 identification 789:802 identification 789:802 identification of the modification site 789:827 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 5 23 from dissociation 1137:1148 arg1 glycopeptide 1097:1108 glycopeptide 1097:1108 glycopeptide 1097:1108 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 10 24 dep https 1818:1822 arg1 //github.com/chpaul/GlycoSeq/ 1824:1852 //github.com/chpaul/GlycoSeq/ 1824:1852 https://github.com/chpaul/GlycoSeq/ 1818:1852 GlycoSeq is released as open source software at https://github.com/chpaul/GlycoSeq/ . 27111718 1 25 theme biomarker 151:159 arg1 analysis 161:168 proteomic biomarker analysis 141:168 proteomic biomarker analysis of human blood samples 141:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 2 26 theme progression 472:482 arg1 markers 453:459 candidate markers 443:459 candidate markers of disease progression 443:482 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 9 27 from glycopeptides 1735:1747 arg1 characterization 1694:1709 the characterization 1690:1709 the characterization of glycan structures in glycopeptides from MS/MS analysis 1690:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 0 28 from Spectra 45:51 arg1 Sequencing 17:26 Automated Glycan Sequencing 0:26 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides 0:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 1 29 theme Mass 80:83 arg1 spectrometry 85:96 Mass spectrometry 80:96 Mass spectrometry 80:96 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 9 30 from structures 1721:1730 arg1 glycopeptides 1735:1747 glycopeptides 1735:1747 glycopeptides from MS/MS analysis 1735:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 9 30 from structures 1721:1730 arg1 analysis 1760:1767 MS/MS analysis 1754:1767 MS/MS analysis 1754:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 0 31 theme Automated 0:8 arg1 Sequencing 17:26 Automated Glycan Sequencing 0:26 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides 0:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 7 32 gly glycopeptides 1400:1412 arg2 glycopeptides 1400:1412 N-linked glycopeptides cell lines 1391:1423 N-linked glycopeptides cell lines acquired from breast cancer patients 1391:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 7 33 theme cell 1414:1417 arg1 lines 1419:1423 N-linked glycopeptides cell lines 1391:1423 N-linked glycopeptides cell lines acquired from breast cancer patients 1391:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 5 34 gly glycopeptide 1097:1108 arg2 glycopeptide 1097:1108 glycopeptide 1097:1108 glycopeptide 1097:1108 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 5 34 gly glycopeptide 1097:1108 arg1 dissociation 1137:1148 its collision-induced dissociation 1115:1148 its collision-induced dissociation 1115:1148 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 4 35 theme site 824:827 arg1 identification 789:802 identification 789:802 identification of the modification site 789:827 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 10 36 theme open 1794:1797 arg1 GlycoSeq 1770:1777 GlycoSeq 1770:1777 GlycoSeq 1770:1777 GlycoSeq is released as open source software at https://github.com/chpaul/GlycoSeq/ . 27111718 10 36 theme open 1794:1797 arg1 software 1806:1813 open source software 1794:1813 open source software 1794:1813 GlycoSeq is released as open source software at https://github.com/chpaul/GlycoSeq/ . 27111718 7 37 theme N-linked 1391:1398 arg1 lines 1419:1423 N-linked glycopeptides cell lines 1391:1423 N-linked glycopeptides cell lines acquired from breast cancer patients 1391:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 2 38 theme multiple 391:398 arg1 diseases 406:413 multiple human diseases 391:413 multiple human diseases 391:413 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 0 39 theme Tandem 33:38 arg1 Spectra 45:51 Tandem Mass Spectra 33:51 Tandem Mass Spectra of N-Linked Glycopeptides 33:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 7 40 theme spectra 1380:1386 arg1 sets 1360:1363 two sets 1356:1363 two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1356:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 7 40 theme spectra 1380:1386 arg1 spectra 1380:1386 tandem mass spectra 1368:1386 tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1368:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 2 41 theme candidate 443:451 arg1 markers 453:459 candidate markers 443:459 candidate markers of disease progression 443:482 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 0 42 theme N-Linked 56:63 arg1 Glycopeptides 65:77 N-Linked Glycopeptides 56:77 N-Linked Glycopeptides 56:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 8 43 theme N-linked 1512:1519 arg1 pathway 1538:1544 the N-linked glycan synthetic pathway 1508:1544 the N-linked glycan synthetic pathway 1508:1544 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 5 44 theme glycopeptide 1097:1108 arg1 spectrum 1162:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 7 45 theme tandem 1368:1373 arg1 spectra 1380:1386 tandem mass spectra 1368:1386 tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1368:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 6 46 dep linkage 1209:1215 arg1 defined 1220:1226 defined 1220:1226 defined by glycan synthetic pathways 1220:1255 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 1 47 theme large 212:216 arg1 availability 218:229 the large availability 208:229 the large availability of informatics tools 208:250 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 5 48 attach present 897:903 arg1 paper 887:891 this paper 882:891 this paper 882:891 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 5 48 attach present 897:903 arg2 we 894:895 we 894:895 we 894:895 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 6 49 theme glycosidic 1198:1207 arg1 linkage 1209:1215 glycosidic linkage 1198:1215 glycosidic linkage as defined by glycan synthetic pathways 1198:1255 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 4 50 theme attached 861:868 arg1 glycans 870:876 the attached glycans 857:876 the attached glycans 857:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 8 51 with consistent 1594:1603 arg1 structures 1638:1647 the manually curated glycan structures 1610:1647 the manually curated glycan structures 1610:1647 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 4 52 from samples 770:776 arg1 structure 844:852 the structure 840:852 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 4 52 from samples 770:776 arg1 identification 728:741 the identification 724:741 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 5 53 theme glycan 1071:1076 arg1 structure 1078:1086 the glycan structure 1067:1086 the glycan structure 1067:1086 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 2 54 theme protein 339:345 arg1 modifications 306:318 the most common protein post-translational modifications 263:318 the most common protein post-translational modifications (PTMs) in mammals 263:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 54 theme protein 339:345 arg1 glycosylation 347:359 protein glycosylation 339:359 protein glycosylation 339:359 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 54 theme protein 339:345 arg1 one 256:258 one 256:258 one 256:258 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 4 55 from glycopeptides 753:765 arg1 samples 770:776 samples 770:776 samples 770:776 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 1 56 theme tools 246:250 arg1 availability 218:229 the large availability 208:229 the large availability of informatics tools 208:250 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 2 57 theme common 272:277 arg1 PTMs 321:324 PTMs 321:324 PTMs 321:324 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 57 theme common 272:277 arg1 modifications 306:318 the most common protein post-translational modifications 263:318 the most common protein post-translational modifications (PTMs) in mammals 263:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 7 58 link N-linked 1391:1398 arg1 lines 1419:1423 N-linked glycopeptides cell lines 1391:1423 N-linked glycopeptides cell lines acquired from breast cancer patients 1391:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 7 59 from tool 1348:1351 arg1 sets 1360:1363 two sets 1356:1363 two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1356:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 7 59 from tool 1348:1351 arg1 spectra 1380:1386 tandem mass spectra 1368:1386 tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1368:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 6 60 theme improbable 1270:1279 arg1 structures 1288:1297 improbable glycan structures 1270:1297 improbable glycan structures 1270:1297 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 9 61 gly glycopeptides 1735:1747 arg2 glycopeptides 1735:1747 glycopeptides 1735:1747 glycopeptides from MS/MS analysis 1735:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 9 61 gly glycopeptides 1735:1747 arg1 analysis 1760:1767 MS/MS analysis 1754:1767 MS/MS analysis 1754:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 1 62 theme human 173:177 arg1 samples 185:191 human blood samples 173:191 human blood samples 173:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 4 63 theme glycosylation 701:713 arg1 characterization 673:688 Complete characterization 664:688 Complete characterization of protein glycosylation 664:713 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 3 64 theme glycosylation-related 576:596 arg1 markers 598:604 glycosylation-related markers 576:604 glycosylation-related markers using existing software 576:628 While mass spectrometry instrumentation has seen advancements in capabilities, discovering glycosylation-related markers using existing software is currently not straightforward. 27111718 1 65 theme samples 185:191 arg1 analysis 161:168 proteomic biomarker analysis 141:168 proteomic biomarker analysis of human blood samples 141:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 6 66 theme synthetic 1238:1246 arg1 pathways 1248:1255 glycan synthetic pathways 1231:1255 glycan synthetic pathways 1231:1255 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 9 67 theme glycan 1714:1719 arg1 structures 1721:1730 glycan structures 1714:1730 glycan structures in glycopeptides from MS/MS analysis 1714:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 2 68 theme post-translational 287:304 arg1 PTMs 321:324 PTMs 321:324 PTMs 321:324 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 2 68 theme post-translational 287:304 arg1 modifications 306:318 the most common protein post-translational modifications 263:318 the most common protein post-translational modifications (PTMs) in mammals 263:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 8 69 theme sequencing 1551:1560 arg1 results 1562:1568 the sequencing results 1547:1568 the sequencing results of GlycoSeq 1547:1580 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 8 69 theme sequencing 1551:1560 arg1 consistent 1594:1603 consistent 1594:1603 consistent 1594:1603 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 4 70 theme Complete 664:671 arg1 characterization 673:688 Complete characterization 664:688 Complete characterization of protein glycosylation 664:713 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 5 71 theme mass 1157:1160 arg1 spectrum 1162:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 a glycopeptide from its collision-induced dissociation tandem mass spectrum 1095:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 1 72 theme routine 111:117 arg1 tool 132:135 a routine experimental tool 109:135 a routine experimental tool for proteomic biomarker analysis of human blood samples 109:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 6 73 theme glycan 1320:1325 arg1 trees 1327:1331 build reasonable glycan trees 1303:1331 build reasonable glycan trees 1303:1331 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 4 74 gly glycopeptides 753:765 arg2 identification 789:802 identification 789:802 identification of the modification site 789:827 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 4 74 gly glycopeptides 753:765 arg2 glycopeptides 753:765 intact glycopeptides 746:765 intact glycopeptides 746:765 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 8 75 theme synthetic 1528:1536 arg1 pathway 1538:1544 the N-linked glycan synthetic pathway 1508:1544 the N-linked glycan synthetic pathway 1508:1544 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 5 76 theme iterated 972:979 arg1 algorithm 999:1007 a heuristic iterated glycan sequencing algorithm 960:1007 a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 960:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 8 77 link N-linked 1512:1519 arg1 pathway 1538:1544 the N-linked glycan synthetic pathway 1508:1544 the N-linked glycan synthetic pathway 1508:1544 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 6 78 theme build 1303:1307 arg1 trees 1327:1331 build reasonable glycan trees 1303:1331 build reasonable glycan trees 1303:1331 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 8 79 theme GlycoSeq 1573:1580 arg1 results 1562:1568 the sequencing results 1547:1568 the sequencing results of GlycoSeq 1547:1580 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 8 79 theme GlycoSeq 1573:1580 arg1 consistent 1594:1603 consistent 1594:1603 consistent 1594:1603 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 0 80 theme Glycopeptides 65:77 arg1 Spectra 45:51 Tandem Mass Spectra 33:51 Tandem Mass Spectra of N-Linked Glycopeptides 33:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 5 81 theme software 930:937 arg1 tool 939:942 an open-source software tool 915:942 an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 915:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 5 81 theme software 930:937 arg1 GlycoSeq 905:912 GlycoSeq 905:912 GlycoSeq 905:912 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 7 82 theme breast 1439:1444 arg1 patients 1453:1460 breast cancer patients 1439:1460 breast cancer patients 1439:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 4 83 from structure 844:852 arg1 samples 770:776 samples 770:776 samples 770:776 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 2 84 theme disease 464:470 arg1 progression 472:482 disease progression 464:482 disease progression 464:482 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 8 85 theme curated 1623:1629 arg1 structures 1638:1647 the manually curated glycan structures 1610:1647 the manually curated glycan structures 1610:1647 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 4 86 theme glycopeptides 753:765 arg1 structure 844:852 the structure 840:852 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 4 86 theme glycopeptides 753:765 arg1 identification 728:741 the identification 724:741 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 5 87 theme prior 1022:1026 arg1 knowledge 1028:1036 prior knowledge 1022:1036 prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 1022:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 2 88 from modifications 306:318 arg1 mammals 330:336 mammals 330:336 mammals 330:336 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 7 89 theme lines 1419:1423 arg1 spectra 1380:1386 tandem mass spectra 1368:1386 tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1368:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 0 90 theme Glycan 10:15 arg1 Sequencing 17:26 Automated Glycan Sequencing 0:26 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides 0:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 5 91 theme sequencing 988:997 arg1 algorithm 999:1007 a heuristic iterated glycan sequencing algorithm 960:1007 a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 960:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 8 92 theme glycan 1631:1636 arg1 structures 1638:1647 the manually curated glycan structures 1610:1647 the manually curated glycan structures 1610:1647 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 7 93 theme glycopeptides 1400:1412 arg1 lines 1419:1423 N-linked glycopeptides cell lines 1391:1423 N-linked glycopeptides cell lines acquired from breast cancer patients 1391:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 2 94 dep alter 382:386 arg1 markers 453:459 candidate markers 443:459 candidate markers of disease progression 443:482 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 4 95 theme modification 811:822 arg1 site 824:827 the modification site 807:827 the modification site 807:827 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 2 96 theme human 400:404 arg1 diseases 406:413 multiple human diseases 391:413 multiple human diseases 391:413 As one of the most common protein post-translational modifications (PTMs) in mammals, protein glycosylation has been observed to alter in multiple human diseases and thus may potentially be candidate markers of disease progression. 27111718 0 97 theme Mass 40:43 arg1 Spectra 45:51 Tandem Mass Spectra 33:51 Tandem Mass Spectra of N-Linked Glycopeptides 33:77 Automated Glycan Sequencing from Tandem Mass Spectra of N-Linked Glycopeptides. 27111718 5 98 theme structure 1078:1086 arg1 elucidation 1052:1062 automated elucidation 1042:1062 automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 1042:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 9 99 from characterization 1694:1709 arg1 glycopeptides 1735:1747 glycopeptides 1735:1747 glycopeptides from MS/MS analysis 1735:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 7 100 theme mass 1375:1378 arg1 spectra 1380:1386 tandem mass spectra 1368:1386 tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients 1368:1460 We tested the tool on two sets of tandem mass spectra of N-linked glycopeptides cell lines acquired from breast cancer patients. 27111718 4 101 theme glycans 870:876 arg1 structure 844:852 the structure 840:852 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 4 101 theme glycans 870:876 arg1 identification 728:741 the identification 724:741 the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans 724:876 Complete characterization of protein glycosylation requires the identification of intact glycopeptides in samples, including identification of the modification site as well as the structure of the attached glycans. 27111718 1 102 theme proteomic 141:149 arg1 analysis 161:168 proteomic biomarker analysis 141:168 proteomic biomarker analysis of human blood samples 141:191 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27111718 9 103 theme structures 1721:1730 arg1 characterization 1694:1709 the characterization 1690:1709 the characterization of glycan structures in glycopeptides from MS/MS analysis 1690:1767 Hence, GlycoSeq is ready to be used for the characterization of glycan structures in glycopeptides from MS/MS analysis. 27111718 6 104 theme linkage 1209:1215 arg1 rules 1189:1193 rules 1189:1193 rules of glycosidic linkage as defined by glycan synthetic pathways 1189:1255 GlycoSeq employs rules of glycosidic linkage as defined by glycan synthetic pathways to eliminate improbable glycan structures and build reasonable glycan trees. 27111718 5 105 theme automated 1042:1050 arg1 elucidation 1052:1062 automated elucidation 1042:1062 automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum 1042:1169 In this paper, we present GlycoSeq, an open-source software tool that implements a heuristic iterated glycan sequencing algorithm coupled with prior knowledge for automated elucidation of the glycan structure within a glycopeptide from its collision-induced dissociation tandem mass spectrum. 27111718 8 106 theme glycan 1521:1526 arg1 pathway 1538:1544 the N-linked glycan synthetic pathway 1508:1544 the N-linked glycan synthetic pathway 1508:1544 After employing enzymatic specificity within the N-linked glycan synthetic pathway, the sequencing results of GlycoSeq were highly consistent with the manually curated glycan structures. 27111718 10 107 theme source 1799:1804 arg1 GlycoSeq 1770:1777 GlycoSeq 1770:1777 GlycoSeq 1770:1777 GlycoSeq is released as open source software at https://github.com/chpaul/GlycoSeq/ . 27111718 10 107 theme source 1799:1804 arg1 software 1806:1813 open source software 1794:1813 open source software 1794:1813 GlycoSeq is released as open source software at https://github.com/chpaul/GlycoSeq/ . 27111718 1 108 theme informatics 234:244 arg1 tools 246:250 informatics tools 234:250 informatics tools 234:250 Mass spectrometry has become a routine experimental tool for proteomic biomarker analysis of human blood samples, partly due to the large availability of informatics tools. 27515086 5 0 from embedding 877:885 arg1 resin 890:894 resin 890:894 resin 890:894 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 0 1 from Localization 55:66 arg1 Tissues 81:87 Mammalian Tissues 71:87 Mammalian Tissues 71:87 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 0 1 from Localization 55:66 arg1 Cells 93:97 Cells 93:97 Cells 93:97 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 1 2 theme mass 236:239 arg1 spectrometry 241:252 mass spectrometry 236:252 mass spectrometry 236:252 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 5 3 theme glycan 787:792 arg1 localization 794:805 glycan localization 787:805 glycan localization 787:805 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 1 4 theme isotope-coded 339:351 arg1 method 389:394 the isotope-coded glycosylation site-specific tagging method 335:394 the isotope-coded glycosylation site-specific tagging method 335:394 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 5 theme glycosylation 353:365 arg1 method 389:394 the isotope-coded glycosylation site-specific tagging method 335:394 the isotope-coded glycosylation site-specific tagging method 335:394 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 6 theme glycan 255:260 arg1 systems 272:278 glycan profiling systems 255:278 glycan profiling systems using lectin microarrays 255:303 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 5 7 theme pre-embedding 741:753 arg1 method 776:781 a pre-embedding electron microscopic method 739:781 a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin 739:894 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 1 8 theme site-specific 367:379 arg1 method 389:394 the isotope-coded glycosylation site-specific tagging method 335:394 the isotope-coded glycosylation site-specific tagging method 335:394 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 3 9 theme diagnosis 539:547 arg1 study 495:499 the study 491:499 the study of glycan function as well as disease diagnosis 491:547 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 1 10 theme glycans 150:156 arg1 study 141:145 the study 137:145 the study of glycans 137:156 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 0 11 theme Lectin 105:110 arg1 Probes 112:117 Lectin Probes 105:117 Lectin Probes 105:117 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 1 12 theme tagging 381:387 arg1 method 389:394 the isotope-coded glycosylation site-specific tagging method 335:394 the isotope-coded glycosylation site-specific tagging method 335:394 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 3 13 from study 495:499 arg1 important 556:564 important 556:564 important 556:564 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 5 14 theme microscopic 764:774 arg1 method 776:781 a pre-embedding electron microscopic method 739:781 a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin 739:894 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 0 15 theme Pre-embedding 0:12 arg1 Method 14:19 Pre-embedding Method 0:19 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 0:118 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 1 16 theme profiling 262:270 arg1 systems 272:278 glycan profiling systems 255:278 glycan profiling systems using lectin microarrays 255:303 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 3 17 from localization 581:592 arg1 cells 620:624 cells 620:624 cells 620:624 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 3 17 from localization 581:592 arg1 tissues 608:614 tissues 608:614 tissues 608:614 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 0 18 theme Electron 24:31 arg1 Microscopy 33:42 Electron Microscopy 24:42 Electron Microscopy 24:42 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 1 19 theme systems 272:278 arg1 development 191:201 the development 187:201 the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis 187:330 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 2 20 theme biological 445:454 arg1 functions 456:464 biological functions 445:464 biological functions 445:464 With these methodologies, glycan structures and biological functions are being elucidated. 27515086 4 21 theme Histochemical 627:639 arg1 methods 641:647 Histochemical methods 627:647 Histochemical methods using lectin probes 627:667 Histochemical methods using lectin probes can localize glycans in the tissues and cells. 27515086 1 22 theme lectin 286:291 arg1 microarrays 293:303 lectin microarrays 286:303 lectin microarrays 286:303 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 4 23 theme lectin 655:660 arg1 probes 662:667 lectin probes 655:667 lectin probes 655:667 Histochemical methods using lectin probes can localize glycans in the tissues and cells. 27515086 3 24 theme glycans 597:603 arg1 localization 581:592 the localization 577:592 the localization of glycans in tissues and cells 577:624 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 2 25 theme glycan 423:428 arg1 structures 430:439 glycan structures 423:439 glycan structures 423:439 With these methodologies, glycan structures and biological functions are being elucidated. 27515086 0 26 theme Microscopy 33:42 arg1 Method 14:19 Pre-embedding Method 0:19 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 0:118 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 5 27 theme electron 755:762 arg1 method 776:781 a pre-embedding electron microscopic method 739:781 a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin 739:894 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 5 28 theme tissue 816:821 arg1 sections 823:830 tissue sections 816:830 tissue sections 816:830 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 3 29 theme glycan 504:509 arg1 function 511:518 glycan function 504:518 glycan function as well as disease diagnosis 504:547 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 0 30 theme Glycan 48:53 arg1 Localization 55:66 Glycan Localization 48:66 Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes 48:117 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 3 31 theme function 511:518 arg1 study 495:499 the study 491:499 the study of glycan function as well as disease diagnosis 491:547 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 3 32 from important 556:564 arg1 study 495:499 the study 491:499 the study of glycan function as well as disease diagnosis 491:547 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 27515086 1 33 theme glycoprotein 310:321 arg1 analysis 323:330 glycoprotein analysis 310:330 glycoprotein analysis 310:330 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 34 theme recent 123:128 arg1 years 130:134 recent years 123:134 recent years 123:134 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 35 theme glycan 206:211 arg1 systems 222:228 glycan analysis systems 206:228 glycan analysis systems using mass spectrometry 206:252 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 36 theme analysis 213:220 arg1 systems 222:228 glycan analysis systems 206:228 glycan analysis systems using mass spectrometry 206:252 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 0 37 theme Mammalian 71:79 arg1 Tissues 81:87 Mammalian Tissues 71:87 Mammalian Tissues 71:87 Pre-embedding Method of Electron Microscopy for Glycan Localization in Mammalian Tissues and Cells Using Lectin Probes. 27515086 5 38 theme prior 868:872 arg1 lectin 861:866 lectin 861:866 lectin prior to embedding in resin 861:894 This chapter describes a pre-embedding electron microscopic method for glycan localization in which tissue sections and cells are incubated with lectin prior to embedding in resin. 27515086 1 39 theme systems 222:228 arg1 development 191:201 the development 187:201 the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis 187:330 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 40 theme analysis 323:330 arg1 development 191:201 the development 187:201 the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis 187:330 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 1 41 gly glycoprotein 310:321 arg1 glycoprotein 310:321 glycoprotein analysis 310:330 glycoprotein analysis 310:330 In recent years, the study of glycans is progressing remarkably by the development of glycan analysis systems using mass spectrometry, glycan profiling systems using lectin microarrays, and glycoprotein analysis by the isotope-coded glycosylation site-specific tagging method. 27515086 3 42 theme disease 531:537 arg1 diagnosis 539:547 disease diagnosis 531:547 glycan function as well as disease diagnosis 504:547 In the study of glycan function as well as disease diagnosis, it is important to examine the localization of glycans in tissues and cells. 26038399 2 0 theme selective 461:469 arg1 pressures 471:479 selective pressures 461:479 selective pressures 461:479 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 7 1 theme detectable 1145:1154 arg1 expression 1156:1165 detectable expression 1145:1165 detectable expression 1145:1165 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 7 2 theme modest 1170:1175 arg1 increase 1177:1184 a modest increase 1168:1184 a modest increase in entry 1168:1193 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 0 3 from Role 4:7 arg1 Glycoprotein 54:65 Ebola Virus Glycoprotein 2 42:67 Ebola Virus Glycoprotein 2 42:67 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 1 4 gly glycoproteins 163:175 arg1 GPs 178:180 GPs 178:180 GPs 178:180 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 4 gly glycoproteins 163:175 arg1 glycoproteins 163:175 viral glycoproteins 157:175 viral glycoproteins (GPs) 157:181 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 5 theme viral 157:161 arg1 GPs 178:180 GPs 178:180 GPs 178:180 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 5 theme viral 157:161 arg1 glycoproteins 163:175 viral glycoproteins 157:175 viral glycoproteins (GPs) 157:181 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 2 6 contain contains 325:332 arg1 N618 405:408 N618 405:408 N618 405:408 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 6 contain contains 325:332 arg2 GP2 306:308 The GP subunit GP2 291:308 The GP subunit GP2 of filoviruses 291:323 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 6 contain contains 325:332 arg2 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 6 contain contains 325:332 arg1 GP2 306:308 The GP subunit GP2 291:308 The GP subunit GP2 of filoviruses 291:323 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 6 contain contains 325:332 arg2 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 6 contain contains 325:332 arg1 N563 396:399 N563 396:399 N563 396:399 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 7 7 theme single 1069:1074 arg1 NGS 1076:1078 a single NGS 1067:1078 a single NGS 1067:1078 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 7 8 from increase 1177:1184 arg1 entry 1189:1193 entry 1189:1193 entry 1189:1193 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 5 9 theme glycan 847:852 arg1 loss 830:833 loss 830:833 loss of the N563 glycan 830:852 However, loss of the N563 glycan enhanced entry by 2-fold and eliminated GP detection by a well-characterized monoclonal antibody KZ52. 26038399 8 10 theme glycans 1311:1317 arg1 importance 1289:1298 the importance 1285:1298 the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization 1285:1398 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 4 11 theme GP 703:704 arg1 expression 706:715 GP expression 703:715 GP expression 703:715 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 12 from impact 724:729 arg1 expression 706:715 GP expression 703:715 GP expression 703:715 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 12 from impact 724:729 arg1 neutralization 743:756 antibody neutralization 734:756 antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP 734:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 5 13 theme monoclonal 931:940 arg1 antibody 942:949 a well-characterized monoclonal antibody KZ52 910:954 a well-characterized monoclonal antibody KZ52 910:954 However, loss of the N563 glycan enhanced entry by 2-fold and eliminated GP detection by a well-characterized monoclonal antibody KZ52. 26038399 8 14 attach removed 1445:1451 arg1 GP1 1458:1460 GP1 1458:1460 GP1 1458:1460 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 8 14 attach removed 1445:1451 arg2 glycans 1428:1434 N-linked glycans 1419:1434 N-linked glycans 1419:1434 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 4 15 theme modest 686:691 arg1 effect 693:698 a modest effect 684:698 a modest effect on GP expression 684:715 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 16 theme antibody 734:741 arg1 neutralization 743:756 antibody neutralization 734:756 antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP 734:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 2 17 link N-linked 357:364 arg1 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 17 link N-linked 357:364 arg1 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 4 18 theme virus 811:815 arg1 GP 817:818 Ebola virus GP 805:818 Ebola virus GP 805:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 5 19 theme GP 894:895 arg1 detection 897:905 GP detection 894:905 GP detection 894:905 However, loss of the N563 glycan enhanced entry by 2-fold and eliminated GP detection by a well-characterized monoclonal antibody KZ52. 26038399 7 20 from GP1 1128:1130 arg1 NGSs 1118:1121 the remaining 16 NGSs 1101:1121 the remaining 16 NGSs from GP1 and GP2 1101:1138 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 2 21 gly glycosylation 366:378 arg2 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 21 gly glycosylation 366:378 arg2 2 334:334 2 334:334 2 334:334 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 21 gly glycosylation 366:378 arg2 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 22 theme subunit 298:304 arg1 GP2 306:308 The GP subunit GP2 291:308 The GP subunit GP2 of filoviruses 291:323 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 23 theme GP 295:296 arg1 GP2 306:308 The GP subunit GP2 291:308 The GP subunit GP2 of filoviruses 291:323 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 24 theme glycosylation 366:378 arg1 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 24 theme glycosylation 366:378 arg1 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 0 25 theme Conserved 12:20 arg1 Glycans 31:37 Conserved N-Linked Glycans 12:37 Conserved N-Linked Glycans 12:37 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 8 26 theme antibody 1376:1383 arg1 neutralization 1385:1398 antibody neutralization 1376:1398 antibody neutralization 1376:1398 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 7 27 from sensitivity 1211:1221 arg1 entry 1189:1193 entry 1189:1193 entry 1189:1193 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 1 28 dep modification 135:146 arg1 involved 187:194 involved 187:194 involved in promoting expression 187:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 28 dep modification 135:146 arg1 found 148:152 found 148:152 found on viral glycoproteins (GPs) 148:181 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 8 29 theme GP2 1307:1309 arg1 glycans 1311:1317 the GP2 glycans 1303:1317 the GP2 glycans 1303:1317 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 1 30 theme antibody 273:280 arg1 evasion 282:288 antibody evasion 273:288 antibody evasion 273:288 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 30 theme antibody 273:280 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 31 from proteases 258:266 arg1 protection 242:251 protection 242:251 protection from proteases 242:266 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 31 from proteases 258:266 arg1 evasion 282:288 antibody evasion 273:288 antibody evasion 273:288 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 31 from proteases 258:266 arg1 attachment 230:239 cellular attachment 221:239 cellular attachment 221:239 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 31 from proteases 258:266 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 4 32 theme glycan 660:665 arg1 Elimination 634:644 Elimination 634:644 Elimination of either GP2 glycan 634:665 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 33 theme GP2 656:658 arg1 glycan 660:665 either GP2 glycan 649:665 either GP2 glycan 649:665 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 0 34 theme Glycans 31:37 arg1 Role 4:7 The Role 0:7 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2 0:67 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 7 35 from GP2 1136:1138 arg1 NGSs 1118:1121 the remaining 16 NGSs 1101:1121 the remaining 16 NGSs from GP1 and GP2 1101:1138 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 0 36 theme N-Linked 22:29 arg1 Glycans 31:37 Conserved N-Linked Glycans 12:37 Conserved N-Linked Glycans 12:37 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 7 37 theme remaining 1105:1113 arg1 NGSs 1118:1121 the remaining 16 NGSs 1101:1121 the remaining 16 NGSs from GP1 and GP2 1101:1138 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 3 38 theme Ebola 606:610 arg1 entry 618:622 Ebola virus entry 606:622 Ebola virus entry 606:622 METHODS: We assessed mutants lacking these glycans for expression and function to understand the role of these sites during Ebola virus entry. 26038399 4 39 theme virus 782:786 arg1 neutralization 743:756 antibody neutralization 734:756 antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP 734:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 7 40 theme antibody 1226:1233 arg1 neutralization 1235:1248 antibody neutralization 1226:1248 antibody neutralization 1226:1248 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 0 41 theme Ebola 42:46 arg1 Glycoprotein 54:65 Ebola Virus Glycoprotein 2 42:67 Ebola Virus Glycoprotein 2 42:67 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 5 42 theme N563 842:845 arg1 glycan 847:852 the N563 glycan 838:852 the N563 glycan 838:852 However, loss of the N563 glycan enhanced entry by 2-fold and eliminated GP detection by a well-characterized monoclonal antibody KZ52. 26038399 3 43 theme virus 612:616 arg1 entry 618:622 Ebola virus entry 606:622 Ebola virus entry 606:622 METHODS: We assessed mutants lacking these glycans for expression and function to understand the role of these sites during Ebola virus entry. 26038399 4 44 theme Ebola 805:809 arg1 GP 817:818 Ebola virus GP 805:818 Ebola virus GP 805:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 45 contain had 680:682 arg2 effect 693:698 a modest effect 684:698 a modest effect on GP expression 684:715 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 45 contain had 680:682 arg2 impact 724:729 no impact 721:729 no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP 721:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 45 contain had 680:682 arg1 Elimination 634:644 Elimination 634:644 Elimination of either GP2 glycan 634:665 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 7 46 theme pronounced 1200:1209 arg1 sensitivity 1211:1221 pronounced sensitivity 1200:1221 pronounced sensitivity to antibody neutralization 1200:1248 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 8 47 theme transduction 1347:1358 arg1 efficiency 1360:1369 transduction efficiency 1347:1369 transduction efficiency 1347:1369 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 8 48 theme N-linked 1419:1426 arg1 glycans 1428:1434 N-linked glycans 1419:1434 N-linked glycans 1419:1434 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 7 49 contain had 1141:1143 arg2 sensitivity 1211:1221 pronounced sensitivity 1200:1221 pronounced sensitivity to antibody neutralization 1200:1248 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 7 49 contain had 1141:1143 arg1 GP 1050:1051 a GP 1048:1051 a GP 1048:1051 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 7 49 contain had 1141:1143 arg2 expression 1156:1165 detectable expression 1145:1165 detectable expression 1145:1165 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 7 49 contain had 1141:1143 arg2 increase 1177:1184 a modest increase 1168:1184 a modest increase in entry 1168:1193 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 2 50 theme filoviruses 313:323 arg1 GP2 306:308 The GP subunit GP2 291:308 The GP subunit GP2 of filoviruses 291:323 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 4 51 from effect 693:698 arg1 expression 706:715 GP expression 703:715 GP expression 703:715 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 4 51 from effect 693:698 arg1 neutralization 743:756 antibody neutralization 734:756 antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP 734:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 7 52 from expression 1156:1165 arg1 entry 1189:1193 entry 1189:1193 entry 1189:1193 Surprisingly, a GP that retained a single NGS at N563, eliminating the remaining 16 NGSs from GP1 and GP2, had detectable expression, a modest increase in entry, and pronounced sensitivity to antibody neutralization. 26038399 3 53 theme sites 593:597 arg1 role 579:582 the role 575:582 the role of these sites during Ebola virus entry 575:622 METHODS: We assessed mutants lacking these glycans for expression and function to understand the role of these sites during Ebola virus entry. 26038399 8 54 theme GP 1322:1323 arg1 expression/structure 1325:1344 GP expression/structure 1322:1344 GP expression/structure 1322:1344 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 1 55 theme common 110:115 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 55 theme common 110:115 arg1 glycosylation 91:103 N-linked glycosylation 82:103 N-linked glycosylation 82:103 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 55 theme common 110:115 arg1 attachment 230:239 cellular attachment 221:239 cellular attachment 221:239 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 55 theme common 110:115 arg1 evasion 282:288 antibody evasion 273:288 antibody evasion 273:288 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 55 theme common 110:115 arg1 protection 242:251 protection 242:251 protection from proteases 242:266 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 55 theme common 110:115 arg1 BACKGROUND 70:79 BACKGROUND 70:79 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 70:289 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 56 theme N-linked 82:89 arg1 BACKGROUND 70:79 BACKGROUND 70:79 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 70:289 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 56 theme N-linked 82:89 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 56 theme N-linked 82:89 arg1 glycosylation 91:103 N-linked glycosylation 82:103 N-linked glycosylation 82:103 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 6 57 theme GP 999:1000 arg1 expression 1002:1011 GP expression 999:1011 GP expression 999:1011 Loss of both sites dramatically decreased GP expression and abolished entry. 26038399 5 58 theme well-characterized 912:929 arg1 antibody 942:949 a well-characterized monoclonal antibody KZ52 910:954 a well-characterized monoclonal antibody KZ52 910:954 However, loss of the N563 glycan enhanced entry by 2-fold and eliminated GP detection by a well-characterized monoclonal antibody KZ52. 26038399 1 59 theme posttranslational 117:133 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 59 theme posttranslational 117:133 arg1 glycosylation 91:103 N-linked glycosylation 82:103 N-linked glycosylation 82:103 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 59 theme posttranslational 117:133 arg1 attachment 230:239 cellular attachment 221:239 cellular attachment 221:239 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 59 theme posttranslational 117:133 arg1 evasion 282:288 antibody evasion 273:288 antibody evasion 273:288 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 59 theme posttranslational 117:133 arg1 protection 242:251 protection 242:251 protection from proteases 242:266 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 59 theme posttranslational 117:133 arg1 BACKGROUND 70:79 BACKGROUND 70:79 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 70:289 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 60 link N-linked 82:89 arg1 BACKGROUND 70:79 BACKGROUND 70:79 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 70:289 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 60 link N-linked 82:89 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 60 link N-linked 82:89 arg1 glycosylation 91:103 N-linked glycosylation 82:103 N-linked glycosylation 82:103 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 8 61 link N-linked 1419:1426 arg1 glycans 1428:1434 N-linked glycans 1419:1434 N-linked glycans 1419:1434 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 4 62 theme stomatitis 771:780 arg1 virus 782:786 vesicular stomatitis virus 761:786 vesicular stomatitis virus pseudotyped with Ebola virus GP 761:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 2 63 theme N-linked 357:364 arg1 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 63 theme N-linked 357:364 arg1 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 8 64 from importance 1289:1298 arg1 neutralization 1385:1398 antibody neutralization 1376:1398 antibody neutralization 1376:1398 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 8 64 from importance 1289:1298 arg1 expression/structure 1325:1344 GP expression/structure 1322:1344 GP expression/structure 1322:1344 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 8 64 from importance 1289:1298 arg1 efficiency 1360:1369 transduction efficiency 1347:1369 transduction efficiency 1347:1369 CONCLUSIONS: Our findings support the importance of the GP2 glycans in GP expression/structure, transduction efficiency, and antibody neutralization, particularly when N-linked glycans are also removed from GP1. 26038399 1 65 theme cellular 221:228 arg1 attachment 230:239 cellular attachment 221:239 cellular attachment 221:239 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 1 65 theme cellular 221:228 arg1 modification 135:146 a common posttranslational modification 108:146 a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression 108:218 BACKGROUND: N-linked glycosylation is a common posttranslational modification found on viral glycoproteins (GPs) and involved in promoting expression, cellular attachment, protection from proteases, and antibody evasion. 26038399 0 66 theme Virus 48:52 arg1 Glycoprotein 54:65 Ebola Virus Glycoprotein 2 42:67 Ebola Virus Glycoprotein 2 42:67 The Role of Conserved N-Linked Glycans on Ebola Virus Glycoprotein 2. 26038399 4 67 theme vesicular 761:769 arg1 virus 782:786 vesicular stomatitis virus 761:786 vesicular stomatitis virus pseudotyped with Ebola virus GP 761:818 RESULTS: Elimination of either GP2 glycan individually had a modest effect on GP expression and no impact on antibody neutralization of vesicular stomatitis virus pseudotyped with Ebola virus GP. 26038399 2 68 theme conserved 347:355 arg1 NGSs 387:390 NGSs 387:390 NGSs 387:390 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 2 68 theme conserved 347:355 arg1 sites 380:384 N-linked glycosylation sites 357:384 2 completely conserved N-linked glycosylation sites (NGSs) 334:391 The GP subunit GP2 of filoviruses contains 2 completely conserved N-linked glycosylation sites (NGSs) at N563 and N618, suggesting that they have been maintained through selective pressures. 26038399 6 69 theme sites 970:974 arg1 Loss 957:960 Loss 957:960 Loss of both sites 957:974 Loss of both sites dramatically decreased GP expression and abolished entry. 28922740 4 0 theme native 856:861 arg1 host 863:866 the native host 852:866 the native host 852:866 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 3 1 theme Escherichia 674:684 arg1 system 691:696 the heterologous Escherichia coli system 657:696 the heterologous Escherichia coli system 657:696 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 3 2 from loss 624:627 arg1 system 691:696 the heterologous Escherichia coli system 657:696 the heterologous Escherichia coli system 657:696 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 7 3 theme Campylobacter 1397:1409 arg1 species 1411:1417 certain Campylobacter species 1389:1417 certain Campylobacter species 1389:1417 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 6 4 theme PglB 1247:1250 arg1 changes 1227:1233 conformational changes 1212:1233 conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide 1212:1318 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 5 5 from jejuni 975:980 arg1 structures 898:907 the PglB crystal structures 881:907 the PglB crystal structures from Campylobacter lari 881:931 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 5 from jejuni 975:980 arg1 Analysis 869:876 Analysis 869:876 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni 869:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 5 from jejuni 975:980 arg1 domain 960:965 the soluble C-terminal domain 937:965 the soluble C-terminal domain from C. jejuni 937:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 3 6 theme acid 558:561 arg1 substitutions 563:575 amino acid substitutions 552:575 amino acid substitutions in the aspartate and lysine residues 552:612 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 8 7 theme substrate 1723:1731 arg1 binding 1668:1674 the binding 1664:1674 the binding of the undecaprenyl-pyrophosphate glycan donor substrate 1664:1731 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 8 8 theme fOS 1775:1777 arg1 production 1779:1788 fOS production 1775:1788 fOS production 1775:1788 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 1 9 theme heptasaccharide 206:220 arg1 transfer 192:199 the transfer 188:199 the transfer of a heptasaccharide from a lipid donor to asparagine 188:253 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 6 10 theme oligosaccharide 1304:1318 arg1 binding 1276:1282 the binding 1272:1282 the binding of the lipid-linked oligosaccharide 1272:1318 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 5 11 theme following 1070:1078 arg1 residues 1088:1095 the two following glycine residues 1062:1095 the two following glycine residues 1062:1095 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 3 12 theme aspartate 584:592 arg1 residues 605:612 the aspartate and lysine residues 580:612 the aspartate and lysine residues 580:612 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 0 13 from function 44:51 arg1 jejuni 110:115 Campylobacter jejuni 96:115 Campylobacter jejuni 96:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 4 14 from levels 808:813 arg1 host 863:866 the native host 852:866 the native host 852:866 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 5 15 theme structures 898:907 arg1 Analysis 869:876 Analysis 869:876 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni 869:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 1 16 from donor 235:239 arg1 transfer 192:199 the transfer 188:199 the transfer of a heptasaccharide from a lipid donor to asparagine 188:253 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 6 17 theme Limited 1166:1172 arg1 experiments 1186:1196 Limited proteolysis experiments 1166:1196 Limited proteolysis experiments 1166:1196 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 3 18 theme lysine 598:603 arg1 residues 605:612 the aspartate and lysine residues 580:612 the aspartate and lysine residues 580:612 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 1 19 theme ≠ 295:295 arg1 P 297:297 X1,2 ≠ P 290:297 X1,2 ≠ P 290:297 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 19 theme ≠ 295:295 arg1 sequon 282:287 the D/E-X1-N-X2-S/T sequon 262:287 the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) 262:298 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 20 theme free 336:339 arg1 fOS 359:361 fOS 359:361 fOS 359:361 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 20 theme free 336:339 arg1 heptasaccharide 317:331 heptasaccharide 317:331 heptasaccharide 317:331 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 20 theme free 336:339 arg1 oligosaccharides 341:356 free oligosaccharides 336:356 free oligosaccharides (fOS) 336:362 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 5 21 theme PglB 885:888 arg1 structures 898:907 the PglB crystal structures 881:907 the PglB crystal structures from Campylobacter lari 881:931 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 22 theme strain 756:761 arg1 complementation 710:724 complementation 710:724 complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles 710:787 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 0 23 theme oligosaccharyltransferase 65:89 arg1 function 44:51 the function 40:51 the function of the PglB oligosaccharyltransferase from Campylobacter jejuni 40:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 5 24 from Analysis 869:876 arg1 jejuni 975:980 C. jejuni 972:980 C. jejuni 972:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 24 from Analysis 869:876 arg1 lari 928:931 lari 928:931 lari 928:931 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 25 theme domain 960:965 arg1 Analysis 869:876 Analysis 869:876 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni 869:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 26 theme mutated 768:774 arg1 alleles 781:787 mutated pglB alleles 768:787 mutated pglB alleles 768:787 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 0 27 theme Campylobacter 96:108 arg1 jejuni 110:115 Campylobacter jejuni 96:115 Campylobacter jejuni 96:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 5 28 theme C. 972:973 arg1 jejuni 975:980 C. jejuni 972:980 C. jejuni 972:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 2 29 theme DGGK 441:444 arg1 motif 446:450 a DGGK motif 439:450 a DGGK motif near the active site of PglB that is conserved among all Campylobacter species 439:529 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 4 30 theme C. 731:732 arg1 strain 756:761 a C. jejuni pglB knock-out strain 729:761 a C. jejuni pglB knock-out strain 729:761 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 2 31 theme available 371:379 arg1 structures 389:398 available crystal structures 371:398 available crystal structures 371:398 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 3 32 dep Escherichia 674:684 arg1 coli 686:689 coli 686:689 coli 686:689 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 5 33 dep subtle 1116:1121 arg1 defined 1129:1135 defined 1129:1135 defined 1129:1135 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 34 theme pglB 741:744 arg1 strain 756:761 a C. jejuni pglB knock-out strain 729:761 a C. jejuni pglB knock-out strain 729:761 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 0 35 theme DGGK 12:15 arg1 essential 26:34 essential 26:34 essential 26:34 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 0 35 theme DGGK 12:15 arg1 motif 17:21 A conserved DGGK motif 0:21 A conserved DGGK motif 0:21 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 5 36 theme lysine 1150:1155 arg1 residue 1157:1163 the lysine residue 1146:1163 the lysine residue 1146:1163 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 37 theme soluble 941:947 arg1 domain 960:965 the soluble C-terminal domain 937:965 the soluble C-terminal domain from C. jejuni 937:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 2 38 theme active 461:466 arg1 PglB 476:479 PglB 476:479 PglB 476:479 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 2 38 theme active 461:466 arg1 site 468:471 the active site 457:471 the active site of PglB 457:479 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 3 39 theme protein 632:638 arg1 glycosylation 640:652 protein glycosylation 632:652 protein glycosylation 632:652 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 2 40 theme sequence 404:411 arg1 alignments 413:422 sequence alignments 404:422 sequence alignments 404:422 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 5 41 theme structural 1016:1025 arg1 role 1027:1030 a particularly important structural role 991:1030 a particularly important structural role for the aspartate residue and the two following glycine residues 991:1095 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 42 theme N-linked 818:825 arg1 glycoproteins 827:839 N-linked glycoproteins 818:839 N-linked glycoproteins 818:839 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 8 43 theme PglB 1569:1572 arg1 structures 1574:1583 the PglB structures 1565:1583 the PglB structures 1565:1583 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 3 44 theme heterologous 661:672 arg1 system 691:696 the heterologous Escherichia coli system 657:696 the heterologous Escherichia coli system 657:696 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 1 45 theme lipid 229:233 arg1 donor 235:239 a lipid donor 227:239 a lipid donor 227:239 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 8 46 theme glycan 1710:1715 arg1 substrate 1723:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 8 47 theme DGGK 1640:1643 arg1 motif 1645:1649 the DGGK motif 1636:1649 the DGGK motif 1636:1649 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 4 48 with complementation 710:724 arg1 alleles 781:787 mutated pglB alleles 768:787 mutated pglB alleles 768:787 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 6 49 link lipid-linked 1291:1302 arg1 oligosaccharide 1304:1318 the lipid-linked oligosaccharide 1287:1318 the lipid-linked oligosaccharide 1287:1318 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 1 50 theme D/E-X1-N-X2-S/T 266:280 arg1 P 297:297 X1,2 ≠ P 290:297 X1,2 ≠ P 290:297 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 50 theme D/E-X1-N-X2-S/T 266:280 arg1 sequon 282:287 the D/E-X1-N-X2-S/T sequon 262:287 the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) 262:298 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 7 51 contain containing 1492:1501 arg1 orthologue 1481:1490 only the orthologue 1472:1490 only the orthologue containing the DGGK motif 1472:1516 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 7 51 contain containing 1492:1501 arg1 active 1521:1526 active 1521:1526 active 1521:1526 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 7 51 contain containing 1492:1501 arg2 motif 1512:1516 the DGGK motif 1503:1516 the DGGK motif 1503:1516 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 7 52 theme PglB 1431:1434 arg1 orthologues 1436:1446 two PglB orthologues 1427:1446 two PglB orthologues 1427:1446 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 4 53 theme glycoproteins 827:839 arg1 fOS 845:847 fOS 845:847 fOS in the native host 845:866 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 4 53 theme glycoproteins 827:839 arg1 levels 808:813 reduced levels 800:813 reduced levels of N-linked glycoproteins 800:839 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 6 54 theme wildtype 1238:1245 arg1 PglB 1247:1250 wildtype PglB 1238:1250 wildtype PglB 1238:1250 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 3 55 theme amino 552:556 arg1 substitutions 563:575 amino acid substitutions 552:575 amino acid substitutions in the aspartate and lysine residues 552:612 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 8 56 theme undecaprenyl-pyrophosphate 1683:1708 arg1 substrate 1723:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 7 57 theme certain 1389:1395 arg1 species 1411:1417 certain Campylobacter species 1389:1417 certain Campylobacter species 1389:1417 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 8 58 theme donor 1717:1721 arg1 substrate 1723:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 the undecaprenyl-pyrophosphate glycan donor substrate 1679:1731 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 4 59 gly glycoproteins 827:839 arg1 glycoproteins 827:839 N-linked glycoproteins 818:839 N-linked glycoproteins 818:839 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 2 60 theme Campylobacter 509:521 arg1 species 523:529 all Campylobacter species 505:529 all Campylobacter species 505:529 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 6 61 theme conformational 1212:1225 arg1 changes 1227:1233 conformational changes 1212:1233 conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide 1212:1318 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 6 62 theme lipid-linked 1291:1302 arg1 oligosaccharide 1304:1318 the lipid-linked oligosaccharide 1287:1318 the lipid-linked oligosaccharide 1287:1318 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 5 63 theme glycine 1080:1086 arg1 residues 1088:1095 the two following glycine residues 1062:1095 the two following glycine residues 1062:1095 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 64 theme crystal 890:896 arg1 structures 898:907 the PglB crystal structures 881:907 the PglB crystal structures from Campylobacter lari 881:931 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 0 65 from jejuni 110:115 arg1 function 44:51 the function 40:51 the function of the PglB oligosaccharyltransferase from Campylobacter jejuni 40:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 0 65 from jejuni 110:115 arg1 oligosaccharyltransferase 65:89 the PglB oligosaccharyltransferase 56:89 the PglB oligosaccharyltransferase from Campylobacter jejuni 56:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 5 66 theme aspartate 1040:1048 arg1 residue 1050:1056 the aspartate residue 1036:1056 the aspartate residue 1036:1056 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 1 67 theme Campylobacter 121:133 arg1 jejuni 135:140 Campylobacter jejuni 121:140 Campylobacter jejuni 121:140 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 6 68 theme proteolysis 1174:1184 arg1 experiments 1186:1196 Limited proteolysis experiments 1166:1196 Limited proteolysis experiments 1166:1196 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 2 69 theme PglB 476:479 arg1 PglB 476:479 PglB 476:479 PglB 476:479 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 2 69 theme PglB 476:479 arg1 site 468:471 the active site 457:471 the active site of PglB 457:479 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 0 70 theme PglB 60:63 arg1 oligosaccharyltransferase 65:89 the PglB oligosaccharyltransferase 56:89 the PglB oligosaccharyltransferase from Campylobacter jejuni 56:115 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 1 71 theme PglB 147:150 arg1 oligosaccharyltransferase 152:176 the PglB oligosaccharyltransferase 143:176 the PglB oligosaccharyltransferase 143:176 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 6 72 theme DGGK 1350:1353 arg1 motif 1355:1359 the DGGK motif 1346:1359 the DGGK motif 1346:1359 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 5 73 theme subtle 1116:1121 arg1 role 1137:1140 a more subtle, less defined role 1109:1140 a more subtle, less defined role for the lysine residue 1109:1163 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 74 theme C-terminal 949:958 arg1 domain 960:965 the soluble C-terminal domain 937:965 the soluble C-terminal domain from C. jejuni 937:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 75 theme knock-out 746:754 arg1 strain 756:761 a C. jejuni pglB knock-out strain 729:761 a C. jejuni pglB knock-out strain 729:761 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 4 76 from fOS 845:847 arg1 host 863:866 the native host 852:866 the native host 852:866 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 4 77 link N-linked 818:825 arg1 glycoproteins 827:839 N-linked glycoproteins 818:839 N-linked glycoproteins 818:839 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 2 78 theme crystal 381:387 arg1 structures 389:398 available crystal structures 371:398 available crystal structures 371:398 Using available crystal structures and sequence alignments, we identified a DGGK motif near the active site of PglB that is conserved among all Campylobacter species. 28922740 0 79 theme conserved 2:10 arg1 essential 26:34 essential 26:34 essential 26:34 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 0 79 theme conserved 2:10 arg1 motif 17:21 A conserved DGGK motif 0:21 A conserved DGGK motif 0:21 A conserved DGGK motif is essential for the function of the PglB oligosaccharyltransferase from Campylobacter jejuni. 28922740 6 80 from changes 1335:1341 arg1 motif 1355:1359 the DGGK motif 1346:1359 the DGGK motif 1346:1359 Limited proteolysis experiments indicate that conformational changes of wildtype PglB that are induced by the binding of the lipid-linked oligosaccharide are altered by changes in the DGGK motif. 28922740 3 81 from substitutions 563:575 arg1 residues 605:612 the aspartate and lysine residues 580:612 the aspartate and lysine residues 580:612 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 1 82 theme X1,2 290:293 arg1 P 297:297 X1,2 ≠ P 290:297 X1,2 ≠ P 290:297 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 1 82 theme X1,2 290:293 arg1 sequon 282:287 the D/E-X1-N-X2-S/T sequon 262:287 the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) 262:298 In Campylobacter jejuni, the PglB oligosaccharyltransferase catalyzes the transfer of a heptasaccharide from a lipid donor to asparagine within the D/E-X1-N-X2-S/T sequon (X1,2 ≠ P) or releases this heptasaccharide as free oligosaccharides (fOS). 28922740 4 83 theme jejuni 734:739 arg1 strain 756:761 a C. jejuni pglB knock-out strain 729:761 a C. jejuni pglB knock-out strain 729:761 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 5 84 theme important 1006:1014 arg1 role 1027:1030 a particularly important structural role 991:1030 a particularly important structural role for the aspartate residue and the two following glycine residues 991:1095 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 4 85 theme reduced 800:806 arg1 levels 808:813 reduced levels 800:813 reduced levels of N-linked glycoproteins 800:839 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 7 86 contain possess 1419:1425 arg1 species 1411:1417 certain Campylobacter species 1389:1417 certain Campylobacter species 1389:1417 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 7 86 contain possess 1419:1425 arg2 orthologues 1436:1446 two PglB orthologues 1427:1446 two PglB orthologues 1427:1446 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 7 87 theme DGGK 1507:1510 arg1 motif 1512:1516 the DGGK motif 1503:1516 the DGGK motif 1503:1516 Related to these findings, certain Campylobacter species possess two PglB orthologues and we demonstrate that only the orthologue containing the DGGK motif is active. 28922740 8 88 theme mutagenesis 1589:1599 arg1 studies 1601:1607 mutagenesis studies 1589:1607 mutagenesis studies 1589:1607 Combining the knowledge gained from the PglB structures and mutagenesis studies, we propose a function for the DGGK motif in affecting the binding of the undecaprenyl-pyrophosphate glycan donor substrate that subsequently influences N-glycan and fOS production. 28922740 3 89 theme glycosylation 640:652 arg1 loss 624:627 loss 624:627 loss of protein glycosylation in the heterologous Escherichia coli system 624:696 We demonstrate that amino acid substitutions in the aspartate and lysine residues result in loss of protein glycosylation in the heterologous Escherichia coli system. 28922740 4 90 theme pglB 776:779 arg1 alleles 781:787 mutated pglB alleles 768:787 mutated pglB alleles 768:787 Similarly, complementation of a C. jejuni pglB knock-out strain with mutated pglB alleles results in reduced levels of N-linked glycoproteins and fOS in the native host. 28922740 5 91 from lari 928:931 arg1 structures 898:907 the PglB crystal structures 881:907 the PglB crystal structures from Campylobacter lari 881:931 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 91 from lari 928:931 arg1 Analysis 869:876 Analysis 869:876 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni 869:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 28922740 5 91 from lari 928:931 arg1 domain 960:965 the soluble C-terminal domain 937:965 the soluble C-terminal domain from C. jejuni 937:980 Analysis of the PglB crystal structures from Campylobacter lari and the soluble C-terminal domain from C. jejuni suggests a particularly important structural role for the aspartate residue and the two following glycine residues, as well as a more subtle, less defined role for the lysine residue. 26378261 7 0 theme NeuAc 1162:1166 arg1 types 1168:1172 the α2-3 NeuAc types 1153:1172 the α2-3 NeuAc types 1153:1172 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 6 1 theme binding 864:870 arg1 site 872:875 the binding style and fucose binding site 835:875 site 872:875 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 2 2 from pattern 202:208 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 2 from pattern 202:208 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 11 3 link iPSC-derived 1761:1772 arg1 cardiomyocytes 1774:1787 iPSC-derived cardiomyocytes 1761:1787 iPSC-derived cardiomyocytes 1761:1787 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 3 4 link iPSC-derived 420:431 arg1 iPSC-CMs 449:456 iPSC-CMs 449:456 iPSC-CMs 449:456 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 4 link iPSC-derived 420:431 arg1 cardiomyocytes 433:446 iPSC-derived cardiomyocytes 420:446 iPSC-derived cardiomyocytes (iPSC-CMs) 420:457 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 10 5 theme cardiomyogenic 1613:1626 arg1 differentiation 1628:1642 cardiomyogenic differentiation 1613:1642 cardiomyogenic differentiation 1613:1642 These findings will prove useful in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 11 6 theme pluripotent 1726:1736 arg1 iPSCs 1750:1754 iPSCs 1750:1754 iPSCs 1750:1754 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 11 6 theme pluripotent 1726:1736 arg1 cells 1743:1747 human induced pluripotent stem cells 1712:1747 human induced pluripotent stem cells (iPSCs) 1712:1755 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 6 7 theme binding 839:845 arg1 style 847:851 the binding style and fucose binding site 835:875 style 847:851 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 6 8 from terminal 895:902 arg1 site 872:875 the binding style and fucose binding site 835:875 site 872:875 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 6 8 from terminal 895:902 arg1 style 847:851 the binding style and fucose binding site 835:875 style 847:851 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 8 9 theme relative 1312:1319 arg1 abundance 1321:1329 relative abundance 1312:1329 relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs 1312:1379 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 4 10 dep chromatography 581:594 arg1 data 696:699 time-of-flight mass-spectrometric data 662:699 time-of-flight mass-spectrometric data 662:699 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 4 10 dep chromatography 581:594 arg1 positions 604:612 elution positions 596:612 elution positions 596:612 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 2 11 theme murine 236:241 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 11 theme murine 236:241 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 7 12 theme terminal 1087:1094 arg1 fucose 1096:1101 the nonreduced terminal fucose 1072:1101 the nonreduced terminal fucose 1072:1101 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 2 13 theme expression 222:231 arg1 pattern 202:208 the pattern 198:208 the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) 198:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 7 14 theme N-acetylglucosamine 1039:1057 arg1 types 1103:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 13 15 theme bisecting 2074:2082 arg1 N-acetylglucosamine 2084:2102 the bisecting N-acetylglucosamine 2070:2102 the bisecting N-acetylglucosamine 2070:2102 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 12 16 located observed 1955:1962 arg2 pattern 1947:1953 the pattern 1943:1953 the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation 1943:2022 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 12 16 located observed 1955:1962 arg1 cardiomyocytes 1973:1986 human cardiomyocytes 1967:1986 human cardiomyocytes 1967:1986 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 11 17 theme cardiomyogenic 1820:1833 arg1 differentiation 1835:1849 cardiomyogenic differentiation 1820:1849 cardiomyogenic differentiation 1820:1849 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 2 18 theme pluripotent 251:261 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 18 theme pluripotent 251:261 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 14 19 theme differentiation 2271:2285 arg1 precision 2243:2251 the directional precision 2227:2251 the directional precision of cardiomyogenic differentiation 2227:2285 The findings of this study will help in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 8 20 located found 1303:1307 arg2 GlcNAc 1259:1264 the bisecting GlcNAc 1245:1264 the bisecting GlcNAc 1245:1264 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 8 20 located found 1303:1307 arg1 abundance 1321:1329 relative abundance 1312:1329 relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs 1312:1379 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 8 20 located found 1303:1307 arg2 structures 1287:1296 the triantennary structures 1270:1296 the triantennary structures 1270:1296 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 7 21 theme terminal 981:988 arg1 process 1004:1010 the terminal glycosylation process 977:1010 the terminal glycosylation process 977:1010 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 13 22 located observed 2058:2065 arg2 differences 2036:2046 Structural differences 2025:2046 Structural differences 2025:2046 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 13 22 located observed 2058:2065 arg1 structures 2125:2134 the triantennary structures 2108:2134 the triantennary structures 2108:2134 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 13 22 located observed 2058:2065 arg1 N-acetylglucosamine 2084:2102 the bisecting N-acetylglucosamine 2070:2102 the bisecting N-acetylglucosamine 2070:2102 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 12 23 theme human 1967:1971 arg1 cardiomyocytes 1973:1986 human cardiomyocytes 1967:1986 human cardiomyocytes 1967:1986 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 14 24 theme directional 2231:2241 arg1 precision 2243:2251 the directional precision 2227:2251 the directional precision of cardiomyogenic differentiation 2227:2285 The findings of this study will help in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 3 25 theme iPSC-derived 420:431 arg1 iPSC-CMs 449:456 iPSC-CMs 449:456 iPSC-CMs 449:456 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 25 theme iPSC-derived 420:431 arg1 cardiomyocytes 433:446 iPSC-derived cardiomyocytes 420:446 iPSC-derived cardiomyocytes (iPSC-CMs) 420:457 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 0 26 theme Structural 0:9 arg1 Changes 11:17 Structural Changes 0:17 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells 0:64 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 1 27 theme Cell-surface 105:116 arg1 glycans 118:124 Cell-surface glycans 105:124 Cell-surface glycans 105:124 Cell-surface glycans vary widely, depending on cell properties. 26378261 8 28 from abundance 1321:1329 arg1 iPSC-CMs 1338:1345 the iPSC-CMs 1334:1345 the iPSC-CMs in comparison with hCMCs or iPSCs 1334:1379 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 11 29 theme iPSC-derived 1761:1772 arg1 cardiomyocytes 1774:1787 iPSC-derived cardiomyocytes 1761:1787 iPSC-derived cardiomyocytes 1761:1787 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 14 30 theme study 2194:2198 arg1 findings 2177:2184 The findings 2173:2184 The findings of this study 2173:2198 The findings of this study will help in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 0 31 theme Pluripotent 43:53 arg1 Cells 60:64 Induced Pluripotent Stem Cells 35:64 Induced Pluripotent Stem Cells 35:64 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 9 32 theme glycosyltransferase-encoding 1405:1432 arg1 MGAT3 1396:1400 MGAT3 1396:1400 MGAT3 1396:1400 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, was higher in iPSCs and iPSC-CMs than in hCMCs. 26378261 9 32 theme glycosyltransferase-encoding 1405:1432 arg1 gene 1434:1437 a glycosyltransferase-encoding gene 1403:1437 a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures 1403:1483 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, was higher in iPSCs and iPSC-CMs than in hCMCs. 26378261 12 33 theme human 1916:1920 arg1 iPSCs 1922:1926 human iPSCs 1916:1926 human iPSCs 1916:1926 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 6 34 theme nonreduced 884:893 arg1 terminal 895:902 the nonreduced terminal 880:902 the nonreduced terminal 880:902 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 7 35 theme N-glycan 958:965 arg1 type 950:953 each type 945:953 each type of N-glycan 945:965 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 12 36 from pattern 1892:1898 arg1 iPSCs 1922:1926 human iPSCs 1916:1926 human iPSCs 1916:1926 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 13 37 theme triantennary 2112:2123 arg1 structures 2125:2134 the triantennary structures 2108:2134 the triantennary structures 2108:2134 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 7 38 theme type 950:953 arg1 Quantitation 929:940 Quantitation 929:940 Quantitation 929:940 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 12 39 theme expression 1881:1890 arg1 pattern 1892:1898 the expression pattern 1877:1898 the expression pattern of N-glycans in human iPSCs 1877:1926 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 4 40 theme laser 634:638 arg1 desorption/ionization 640:660 matrix-assisted laser desorption/ionization 618:660 matrix-assisted laser desorption/ionization 618:660 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 10 41 theme directional 1588:1598 arg1 precision 1600:1608 the directional precision 1584:1608 the directional precision of cardiomyogenic differentiation 1584:1642 These findings will prove useful in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 11 42 theme induced 1718:1724 arg1 iPSCs 1750:1754 iPSCs 1750:1754 iPSCs 1750:1754 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 11 42 theme induced 1718:1724 arg1 cells 1743:1747 human induced pluripotent stem cells 1712:1747 human induced pluripotent stem cells (iPSCs) 1712:1755 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 7 43 theme α2-3 1157:1160 arg1 types 1168:1172 the α2-3 NeuAc types 1153:1172 the α2-3 NeuAc types 1153:1172 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 9 44 theme GlcNAc 1467:1472 arg1 structures 1474:1483 the bisecting GlcNAc structures 1453:1483 the bisecting GlcNAc structures 1453:1483 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, was higher in iPSCs and iPSC-CMs than in hCMCs. 26378261 4 45 theme liquid 574:579 arg1 chromatography 581:594 high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data 557:699 chromatography 581:594 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 5 46 theme isolated 708:715 arg1 N-glycans 717:725 52 isolated N-glycans 705:725 52 isolated N-glycans 705:725 Of 52 isolated N-glycans, the structures of 38 were clearly identified. 26378261 6 47 theme fucose 857:862 arg1 site 872:875 the binding style and fucose binding site 835:875 site 872:875 In addition, 11 structures were partially identified because the binding style and fucose binding site at the nonreduced terminal could not be identified. 26378261 4 48 theme elution 596:602 arg1 positions 604:612 elution positions 596:612 elution positions 596:612 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 10 49 theme differentiation 1628:1642 arg1 precision 1600:1608 the directional precision 1584:1608 the directional precision of cardiomyogenic differentiation 1584:1642 These findings will prove useful in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 7 50 theme exposed 1132:1138 arg1 galactose 1140:1148 the exposed galactose 1128:1148 the exposed galactose 1128:1148 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 8 51 theme triantennary 1274:1285 arg1 structures 1287:1296 the triantennary structures 1270:1296 the triantennary structures 1270:1296 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 11 52 theme stem 1738:1741 arg1 iPSCs 1750:1754 iPSCs 1750:1754 iPSCs 1750:1754 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 11 52 theme stem 1738:1741 arg1 cells 1743:1747 human induced pluripotent stem cells 1712:1747 human induced pluripotent stem cells (iPSCs) 1712:1755 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 2 53 theme cardiac 309:315 arg1 tissue 317:322 the cardiac tissue 305:322 the cardiac tissue 305:322 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 5 54 theme 38 746:747 arg1 structures 732:741 the structures 728:741 the structures of 38 728:747 Of 52 isolated N-glycans, the structures of 38 were clearly identified. 26378261 4 55 theme time-of-flight 662:675 arg1 data 696:699 time-of-flight mass-spectrometric data 662:699 time-of-flight mass-spectrometric data 662:699 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 7 56 theme fucose 1096:1101 arg1 types 1103:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 2 57 theme induced 243:249 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 57 theme induced 243:249 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 8 58 theme bisecting 1249:1257 arg1 GlcNAc 1259:1264 the bisecting GlcNAc 1245:1264 the bisecting GlcNAc 1245:1264 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 7 59 theme nonreduced 1076:1085 arg1 fucose 1096:1101 the nonreduced terminal fucose 1072:1101 the nonreduced terminal fucose 1072:1101 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 3 60 attach isolated 393:400 arg1 cardiomyocytes 433:446 iPSC-derived cardiomyocytes 420:446 iPSC-derived cardiomyocytes (iPSC-CMs) 420:457 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 60 attach isolated 393:400 arg1 cardiomyocytes 470:483 human cardiomyocytes 464:483 human cardiomyocytes (hCMCs) 464:491 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 60 attach isolated 393:400 arg1 iPSC-CMs 449:456 iPSC-CMs 449:456 iPSC-CMs 449:456 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 60 attach isolated 393:400 arg2 N-glycans 378:386 N-glycans 378:386 N-glycans 378:386 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 60 attach isolated 393:400 arg1 hCMCs 486:490 hCMCs 486:490 hCMCs 486:490 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 60 attach isolated 393:400 arg1 iPSCs 413:417 human iPSCs 407:417 human iPSCs 407:417 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 13 61 theme Structural 2025:2034 arg1 differences 2036:2046 Structural differences 2025:2046 Structural differences 2025:2046 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 2 62 theme stem 263:266 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 62 theme stem 263:266 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 4 63 theme mass-spectrometric 677:694 arg1 data 696:699 time-of-flight mass-spectrometric data 662:699 time-of-flight mass-spectrometric data 662:699 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 1 64 theme cell 152:155 arg1 properties 157:166 cell properties 152:166 cell properties 152:166 Cell-surface glycans vary widely, depending on cell properties. 26378261 0 65 from Changes 11:17 arg1 Cells 60:64 Induced Pluripotent Stem Cells 35:64 Induced Pluripotent Stem Cells 35:64 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 0 65 from Changes 11:17 arg1 N-Glycans 22:30 N-Glycans 22:30 N-Glycans 22:30 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 8 66 from iPSC-CMs 1338:1345 arg1 comparison 1350:1359 comparison 1350:1359 comparison with hCMCs or iPSCs 1350:1379 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 14 67 theme cardiomyogenic 2256:2269 arg1 differentiation 2271:2285 cardiomyogenic differentiation 2256:2285 cardiomyogenic differentiation 2256:2285 The findings of this study will help in understanding the directional precision of cardiomyogenic differentiation in vitro. 26378261 7 68 theme glycosylation 990:1002 arg1 process 1004:1010 the terminal glycosylation process 977:1010 the terminal glycosylation process 977:1010 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 2 69 from expression 222:231 arg1 iPSCs 275:279 iPSCs 275:279 iPSCs 275:279 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 69 from expression 222:231 arg1 cells 268:272 murine induced pluripotent stem cells 236:272 murine induced pluripotent stem cells (iPSCs) 236:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 7 70 theme exposed 1031:1037 arg1 types 1103:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types 1027:1107 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 7 71 theme cardiomyogenic 1204:1217 arg1 differentiation 1219:1233 cardiomyogenic differentiation 1204:1233 cardiomyogenic differentiation 1204:1233 Quantitation of each type of N-glycan, based on the terminal glycosylation process, revealed that the exposed N-acetylglucosamine (GlcNAc) and the nonreduced terminal fucose types decreased, whereas the exposed galactose or the α2-3 NeuAc types increased in the iPSCs during cardiomyogenic differentiation. 26378261 12 72 theme cardiomyogenic 1993:2006 arg1 differentiation 2008:2022 cardiomyogenic differentiation 1993:2022 cardiomyogenic differentiation 1993:2022 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 4 73 theme high-performance 557:572 arg1 chromatography 581:594 high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data 557:699 chromatography 581:594 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 8 74 with comparison 1350:1359 arg1 hCMCs 1366:1370 hCMCs 1366:1370 hCMCs 1366:1370 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 8 74 with comparison 1350:1359 arg1 iPSCs 1375:1379 iPSCs 1375:1379 iPSCs 1375:1379 However, the bisecting GlcNAc and the triantennary structures were found in relative abundance in the iPSC-CMs in comparison with hCMCs or iPSCs. 26378261 3 75 theme human 464:468 arg1 hCMCs 486:490 hCMCs 486:490 hCMCs 486:490 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 3 75 theme human 464:468 arg1 cardiomyocytes 470:483 human cardiomyocytes 464:483 human cardiomyocytes (hCMCs) 464:491 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 0 76 theme Induced 35:41 arg1 Cells 60:64 Induced Pluripotent Stem Cells 35:64 Induced Pluripotent Stem Cells 35:64 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 9 77 theme MGAT3 1396:1400 arg1 Expression 1382:1391 Expression 1382:1391 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, 1382:1484 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, was higher in iPSCs and iPSC-CMs than in hCMCs. 26378261 2 78 from cells 268:272 arg1 pattern 202:208 the pattern 198:208 the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) 198:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 13 79 theme cardiomyogenic 2141:2154 arg1 differentiation 2156:2170 cardiomyogenic differentiation 2141:2170 cardiomyogenic differentiation 2141:2170 Structural differences were also observed in the bisecting N-acetylglucosamine and the triantennary structures upon cardiomyogenic differentiation. 26378261 0 80 theme Stem 55:58 arg1 Cells 60:64 Induced Pluripotent Stem Cells 35:64 Induced Pluripotent Stem Cells 35:64 Structural Changes in N-Glycans on Induced Pluripotent Stem Cells Differentiating Toward Cardiomyocytes. 26378261 3 81 theme human 407:411 arg1 iPSCs 413:417 human iPSCs 407:417 human iPSCs 407:417 In this study, N-glycans were isolated from human iPSCs, iPSC-derived cardiomyocytes (iPSC-CMs), and human cardiomyocytes (hCMCs). 26378261 4 82 theme mapping 530:536 arg1 technique 538:546 a mapping technique 528:546 a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data 528:699 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 2 83 theme N-glycan 213:220 arg1 expression 222:231 N-glycan expression 213:231 N-glycan expression on murine induced pluripotent stem cells (iPSCs) 213:280 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 2 84 theme cardiomyogenic 331:344 arg1 differentiation 346:360 cardiomyogenic differentiation 331:360 cardiomyogenic differentiation 331:360 Previously, we reported that the pattern of N-glycan expression on murine induced pluripotent stem cells (iPSCs) changed toward that of the cardiac tissue during cardiomyogenic differentiation. 26378261 12 85 theme N-glycans 1903:1911 arg1 pattern 1892:1898 the expression pattern 1877:1898 the expression pattern of N-glycans in human iPSCs 1877:1926 This shows that the expression pattern of N-glycans in human iPSCs changed toward the pattern observed in human cardiomyocytes upon cardiomyogenic differentiation. 26378261 4 86 theme matrix-assisted 618:632 arg1 desorption/ionization 640:660 matrix-assisted laser desorption/ionization 618:660 matrix-assisted laser desorption/ionization 618:660 Their structures were analyzed by a mapping technique based on high-performance liquid chromatography elution positions and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometric data. 26378261 11 87 theme human 1712:1716 arg1 iPSCs 1750:1754 iPSCs 1750:1754 iPSCs 1750:1754 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 11 87 theme human 1712:1716 arg1 cells 1743:1747 human induced pluripotent stem cells 1712:1747 human induced pluripotent stem cells (iPSCs) 1712:1755 SIGNIFICANCE: This study focused on N-glycans produced in human induced pluripotent stem cells (iPSCs) and iPSC-derived cardiomyocytes to investigate their change on cardiomyogenic differentiation in vitro. 26378261 9 88 theme bisecting 1457:1465 arg1 structures 1474:1483 the bisecting GlcNAc structures 1453:1483 the bisecting GlcNAc structures 1453:1483 Expression of MGAT3, a glycosyltransferase-encoding gene that produces the bisecting GlcNAc structures, was higher in iPSCs and iPSC-CMs than in hCMCs. 26156869 5 0 theme glycan 1184:1189 arg1 pool 1191:1194 the glycan pool 1180:1194 the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 1180:1240 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 3 1 theme therapeutic 710:720 arg1 denosumab 791:799 denosumab 791:799 denosumab 791:799 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 1 theme therapeutic 710:720 arg1 cetuximab 752:760 cetuximab 752:760 cetuximab 752:760 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 1 theme therapeutic 710:720 arg1 antibodies 733:742 therapeutic monoclonal antibodies 710:742 therapeutic monoclonal antibodies (mAbs) 710:749 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 1 theme therapeutic 710:720 arg1 adalimumab 763:772 adalimumab 763:772 adalimumab 763:772 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 1 theme therapeutic 710:720 arg1 mAbs 745:748 mAbs 745:748 mAbs 745:748 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 1 theme therapeutic 710:720 arg1 panitumumab 775:785 panitumumab 775:785 panitumumab 775:785 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 4 2 theme oligomannose 1037:1048 arg1 structures 1050:1059 hybrid and oligomannose structures 1026:1059 structures 1050:1059 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 1 3 theme defense 250:256 arg1 responses 258:266 defense responses 250:266 defense responses 250:266 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 0 4 theme glycoform 79:87 arg1 selectivity 89:99 different glycoform selectivity 69:99 different glycoform selectivity 69:99 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 3 5 theme EndoS2 685:690 arg1 activities 661:670 the activities 657:670 the activities of EndoS and EndoS2 657:690 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 2 6 dep immunoglobulin 460:473 arg1 G. 475:476 G. 475:476 G. 475:476 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 5 7 theme cetuximab 1199:1207 arg1 pool 1191:1194 the glycan pool 1180:1194 the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 1180:1240 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 2 8 theme activities 547:556 arg1 comparison 517:526 comparison 517:526 comparison of the hydrolyzing activities 517:556 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 8 theme activities 547:556 arg1 characterization 496:511 detailed characterization 487:511 detailed characterization 487:511 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 5 9 theme chromatography 1148:1161 arg1 profiles 1168:1175 ultra-high-performance liquid chromatography (LC) profiles 1118:1175 ultra-high-performance liquid chromatography (LC) profiles 1118:1175 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 2 10 link N-linked 434:441 arg1 glycans 443:449 N-linked glycans 434:449 N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed 434:580 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 5 11 theme hybrid 1272:1277 arg1 glycans 1296:1302 hybrid and oligomannose glycans 1272:1302 glycans 1296:1302 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 2 12 theme hydrolyzing 535:545 arg1 activities 547:556 the hydrolyzing activities 531:556 the hydrolyzing activities 531:556 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 3 13 theme monoclonal 722:731 arg1 denosumab 791:799 denosumab 791:799 denosumab 791:799 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 13 theme monoclonal 722:731 arg1 cetuximab 752:760 cetuximab 752:760 cetuximab 752:760 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 13 theme monoclonal 722:731 arg1 antibodies 733:742 therapeutic monoclonal antibodies 710:742 therapeutic monoclonal antibodies (mAbs) 710:749 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 13 theme monoclonal 722:731 arg1 adalimumab 763:772 adalimumab 763:772 adalimumab 763:772 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 13 theme monoclonal 722:731 arg1 mAbs 745:748 mAbs 745:748 mAbs 745:748 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 3 13 theme monoclonal 722:731 arg1 panitumumab 775:785 panitumumab 775:785 panitumumab 775:785 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 1 14 gly glycoproteins 187:199 arg1 glycoproteins 187:199 glycoproteins 187:199 glycoproteins of the human immune system 187:226 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 7 15 theme Fc-glycosylation 1644:1659 arg1 site 1661:1664 the Fc-glycosylation site 1640:1664 the Fc-glycosylation site on therapeutic mAbs 1640:1684 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 5 16 theme EndoS 1345:1349 arg1 chromatogram 1351:1362 the EndoS chromatogram 1341:1362 the EndoS chromatogram 1341:1362 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 3 17 theme antibodies 733:742 arg1 selection 697:705 a selection 695:705 a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab 695:799 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 2 18 dep glycans 443:449 arg1 performed 572:580 performed 572:580 have not been performed 558:580 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 7 19 theme different 1614:1622 arg1 glycoforms 1624:1633 different glycoforms 1614:1633 different glycoforms 1614:1633 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 6 20 theme Fc 1535:1536 arg1 fragments 1538:1546 the Fc fragments 1531:1546 the Fc fragments of the selected mAbs 1531:1567 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 0 21 theme therapeutic 41:51 arg1 antibodies 53:62 therapeutic antibodies 41:62 therapeutic antibodies with different glycoform selectivity 41:99 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 2 22 theme detailed 487:494 arg1 characterization 496:511 detailed characterization 487:511 detailed characterization 487:511 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 5 23 theme ultra-high-performance 1118:1139 arg1 LC 1164:1165 LC 1164:1165 LC 1164:1165 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 5 23 theme ultra-high-performance 1118:1139 arg1 chromatography 1148:1161 ultra-high-performance liquid chromatography 1118:1161 ultra-high-performance liquid chromatography (LC) profiles 1118:1175 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 6 24 theme IdeS 1443:1446 arg1 protease 1448:1455 the IdeS protease 1439:1455 the IdeS protease 1439:1455 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 2 25 theme human 454:458 arg1 immunoglobulin 460:473 human immunoglobulin G. 454:476 human immunoglobulin G. 454:476 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 0 26 from Fc-glycans 27:36 arg1 antibodies 53:62 therapeutic antibodies 41:62 therapeutic antibodies with different glycoform selectivity 41:99 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 2 27 from pyogenes 368:375 arg1 endoglycosidases 313:328 Two endoglycosidases 309:328 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, 309:394 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 4 28 dep enzymes 967:973 arg1 both 958:961 both 958:961 both 958:961 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 5 29 from chromatogram 1351:1362 arg1 missing 1330:1336 missing 1330:1336 missing 1330:1336 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 5 30 theme liquid 1141:1146 arg1 LC 1164:1165 LC 1164:1165 LC 1164:1165 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 5 30 theme liquid 1141:1146 arg1 chromatography 1148:1161 ultra-high-performance liquid chromatography 1118:1161 ultra-high-performance liquid chromatography (LC) profiles 1118:1175 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 6 31 with combination 1422:1432 arg1 protease 1448:1455 the IdeS protease 1439:1455 the IdeS protease 1439:1455 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 3 32 theme present 590:596 arg1 study 598:602 the present study 586:602 the present study 586:602 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 7 33 theme mannose 1745:1751 arg1 content 1753:1759 high mannose content 1740:1759 high mannose content 1740:1759 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 0 34 theme rapid 121:125 arg1 quantification 127:140 rapid quantification 121:140 rapid quantification of high-mannose glycans 121:164 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 7 35 theme content 1753:1759 arg1 quantification 1722:1735 rapid quantification 1716:1735 rapid quantification of high mannose content 1716:1759 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 6 36 theme mannose 1512:1518 arg1 content 1520:1526 high mannose content 1507:1526 high mannose content 1507:1526 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 2 37 theme N-linked 434:441 arg1 glycans 443:449 N-linked glycans 434:449 N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed 434:580 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 1 38 theme bacterial 288:296 arg1 pathogens 298:306 bacterial pathogens 288:306 bacterial pathogens 288:306 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 4 39 theme ionization 917:926 arg1 time 928:931 matrix-assisted laser desorption ionization time 884:931 matrix-assisted laser desorption ionization time of flight 884:941 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 6 40 theme high 1507:1510 arg1 content 1520:1526 high mannose content 1507:1526 high mannose content 1507:1526 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 1 41 theme human 208:212 arg1 system 221:226 the human immune system 204:226 the human immune system 204:226 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 5 42 theme pool 1191:1194 arg1 comparison 1104:1113 A comparison 1102:1113 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 1102:1240 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 4 43 theme flight 936:941 arg1 time 928:931 matrix-assisted laser desorption ionization time 884:931 matrix-assisted laser desorption ionization time of flight 884:941 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 6 44 theme LC 1474:1475 arg1 method 1488:1493 a LC separation method 1472:1493 a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs 1472:1567 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 1 45 theme immune 214:219 arg1 system 221:226 the human immune system 204:226 the human immune system 204:226 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 4 46 theme complex 983:989 arg1 glycans 991:997 complex glycans 983:997 complex glycans 983:997 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 7 47 from site 1661:1664 arg1 mAbs 1681:1684 therapeutic mAbs 1669:1684 therapeutic mAbs 1669:1684 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 1 48 theme system 221:226 arg1 glycoproteins 187:199 glycoproteins 187:199 glycoproteins of the human immune system 187:226 Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. 26156869 0 49 theme glycans 158:164 arg1 quantification 127:140 rapid quantification 121:140 rapid quantification of high-mannose glycans 121:164 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 7 50 used used 1707:1710 arg2 this 1695:1698 this 1695:1698 this 1695:1698 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 7 51 theme rapid 1716:1720 arg1 quantification 1722:1735 rapid quantification 1716:1735 rapid quantification of high mannose content 1716:1759 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 2 52 theme Streptococcus 354:366 arg1 EndoS2 388:393 EndoS2 388:393 EndoS2 388:393 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 52 theme Streptococcus 354:366 arg1 EndoS 378:382 EndoS 378:382 EndoS 378:382 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 52 theme Streptococcus 354:366 arg1 pyogenes 368:375 the human pathogen Streptococcus pyogenes 335:375 the human pathogen Streptococcus pyogenes 335:375 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 53 theme immunoglobulin 460:473 arg1 glycans 443:449 N-linked glycans 434:449 N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed 434:580 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 4 54 theme matrix-assisted 884:898 arg1 ionization 917:926 matrix-assisted laser desorption ionization 884:926 matrix-assisted laser desorption ionization time of flight 884:941 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 0 55 theme high-mannose 145:156 arg1 glycans 158:164 high-mannose glycans 145:164 high-mannose glycans 145:164 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 3 56 theme EndoS 675:679 arg1 activities 661:670 the activities 657:670 the activities of EndoS and EndoS2 657:690 In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosumab. 26156869 2 57 theme pathogen 345:352 arg1 EndoS2 388:393 EndoS2 388:393 EndoS2 388:393 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 57 theme pathogen 345:352 arg1 EndoS 378:382 EndoS 378:382 EndoS 378:382 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 57 theme pathogen 345:352 arg1 pyogenes 368:375 the human pathogen Streptococcus pyogenes 335:375 the human pathogen Streptococcus pyogenes 335:375 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 5 58 theme profiles 1168:1175 arg1 comparison 1104:1113 A comparison 1102:1113 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 1102:1240 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 4 59 theme desorption 906:915 arg1 ionization 917:926 matrix-assisted laser desorption ionization 884:926 matrix-assisted laser desorption ionization time of flight 884:941 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 0 60 used used 112:115 arg2 EndoS 0:4 EndoS 0:4 EndoS 0:4 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 2 61 theme human 339:343 arg1 EndoS2 388:393 EndoS2 388:393 EndoS2 388:393 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 61 theme human 339:343 arg1 EndoS 378:382 EndoS 378:382 EndoS 378:382 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 2 61 theme human 339:343 arg1 pyogenes 368:375 the human pathogen Streptococcus pyogenes 335:375 the human pathogen Streptococcus pyogenes 335:375 Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. 26156869 4 62 theme greater 1066:1072 arg1 extent 1074:1079 a greater extent 1064:1079 a greater extent 1064:1079 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 4 63 theme laser 900:904 arg1 ionization 917:926 matrix-assisted laser desorption ionization 884:926 matrix-assisted laser desorption ionization time of flight 884:941 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 6 64 theme glycoform 1396:1404 arg1 selectivity 1406:1416 glycoform selectivity 1396:1416 glycoform selectivity 1396:1416 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 0 65 with antibodies 53:62 arg1 selectivity 89:99 different glycoform selectivity 69:99 different glycoform selectivity 69:99 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 7 66 theme therapeutic 1669:1679 arg1 mAbs 1681:1684 therapeutic mAbs 1669:1684 therapeutic mAbs 1669:1684 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 6 67 theme separation 1477:1486 arg1 method 1488:1493 a LC separation method 1472:1493 a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs 1472:1567 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 0 68 theme different 69:77 arg1 selectivity 89:99 different glycoform selectivity 69:99 different glycoform selectivity 69:99 EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans. 26156869 5 69 from missing 1330:1336 arg1 chromatogram 1351:1362 the EndoS chromatogram 1341:1362 the EndoS chromatogram 1341:1362 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 6 70 theme selected 1555:1562 arg1 mAbs 1564:1567 the selected mAbs 1551:1567 the selected mAbs 1551:1567 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 7 71 gly Fc-glycosylation 1644:1659 arg2 site 1661:1664 the Fc-glycosylation site 1640:1664 the Fc-glycosylation site on therapeutic mAbs 1640:1684 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 26156869 5 72 theme oligomannose 1283:1294 arg1 glycans 1296:1302 hybrid and oligomannose glycans 1272:1302 glycans 1296:1302 A comparison of ultra-high-performance liquid chromatography (LC) profiles of the glycan pool of cetuximab hydrolyzed with EndoS and EndoS2 showed that EndoS2 hydrolyzed hybrid and oligomannose glycans, whereas these peaks were missing in the EndoS chromatogram. 26156869 6 73 used utilized 1368:1375 arg2 We 1365:1366 We 1365:1366 We 1365:1366 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 6 74 from difference 1382:1391 arg1 selectivity 1406:1416 glycoform selectivity 1396:1416 glycoform selectivity 1396:1416 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 6 74 from difference 1382:1391 arg1 combination 1422:1432 combination 1422:1432 combination with the IdeS protease 1422:1455 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 4 75 theme hybrid 1026:1031 arg1 structures 1050:1059 hybrid and oligomannose structures 1026:1059 structures 1050:1059 By analyzing the glycans hydrolyzed by EndoS and EndoS2 from the antibodies using matrix-assisted laser desorption ionization time of flight, we found that both the enzymes cleaved complex glycans and that EndoS2 hydrolyzed hybrid and oligomannose structures to a greater extent compared with EndoS. 26156869 6 76 theme mAbs 1564:1567 arg1 fragments 1538:1546 the Fc fragments 1531:1546 the Fc fragments of the selected mAbs 1531:1567 We utilized this difference in glycoform selectivity, in combination with the IdeS protease, and developed a LC separation method to quantify high mannose content in the Fc fragments of the selected mAbs. 26156869 7 77 theme high 1740:1743 arg1 content 1753:1759 high mannose content 1740:1759 high mannose content 1740:1759 We conclude that EndoS and EndoS2 hydrolyze different glycoforms from the Fc-glycosylation site on therapeutic mAbs and that this can be used for rapid quantification of high mannose content. 28717478 7 0 theme cell 1374:1377 arg1 epitope 1379:1385 the T cell epitope 1368:1385 the T cell epitope 1368:1385 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 9 1 gly glycopeptides 1747:1759 arg2 glycopeptides 1747:1759 the glycopeptides 1743:1759 the glycopeptides 1743:1759 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 0 2 dep synthesis 27:35 arg1 targeting 68:76 targeting 68:76 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 0:136 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 3 3 theme cell 619:622 arg1 epitope 624:630 a well-characterised T cell epitope 596:630 a well-characterised T cell epitope 596:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 7 4 theme high-mannose 1310:1321 arg1 structures 1332:1341 high-mannose N-glycan structures 1310:1341 high-mannose N-glycan structures 1310:1341 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 2 5 theme Mannose-terminated 371:388 arg1 oligosaccharides 399:414 Mannose-terminated N-glycan oligosaccharides 371:414 Mannose-terminated N-glycan oligosaccharides 371:414 Mannose-terminated N-glycan oligosaccharides, produced by either total or semi-synthesis, were converted into oxazoline donor substrates. 28717478 6 6 theme non-glycosylated 1262:1277 arg1 control 1279:1285 the non-glycosylated control 1258:1285 the non-glycosylated control 1258:1285 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 6 7 gly non-glycosylated 1262:1277 arg1 control 1279:1285 the non-glycosylated control 1258:1285 the non-glycosylated control 1258:1285 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 5 8 theme antigen 983:989 arg1 APCs 1009:1012 APCs 1009:1012 APCs 1009:1012 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 5 8 theme antigen 983:989 arg1 cells 1002:1006 human antigen presenting cells 977:1006 human antigen presenting cells (APCs) 977:1013 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 8 9 theme cell 1560:1563 arg1 activation 1565:1574 T cell activation 1558:1574 T cell activation 1558:1574 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 3 10 theme phase 712:716 arg1 synthesis 726:734 solid phase peptide synthesis 706:734 solid phase peptide synthesis 706:734 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 7 11 theme clone 1460:1464 arg1 activation 1437:1446 activation 1437:1446 activation of a T cell clone 1437:1464 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 7 12 theme T 1372:1372 arg1 epitope 1379:1385 the T cell epitope 1368:1385 the T cell epitope 1368:1385 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 1 13 theme phase 163:167 arg1 synthesis 177:185 solid phase peptide synthesis 157:185 solid phase peptide synthesis 157:185 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 4 14 theme enzymatic 786:794 arg1 glycosylation 796:808 High-yielding enzymatic glycosylation 772:808 High-yielding enzymatic glycosylation 772:808 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 0 15 theme vaccine 90:96 arg1 candidates 98:107 putative vaccine candidates 81:107 putative vaccine candidates 81:107 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 1 16 theme synthesis 177:185 arg1 combination 142:152 The combination 138:152 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 138:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 16 theme synthesis 177:185 arg1 method 290:295 a powerful convergent synthetic method 258:295 a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures 258:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 7 17 theme single 1348:1353 arg1 site 1355:1358 a single site 1346:1358 a single site outside the T cell epitope 1346:1385 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 0 18 theme presenting 120:129 arg1 cells 131:135 antigen presenting cells 112:135 antigen presenting cells 112:135 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 0 19 theme glycopeptides 53:65 arg1 synthesis 27:35 Convergent chemo-enzymatic synthesis 0:35 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 0:136 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 7 20 theme cell 1455:1458 arg1 clone 1460:1464 a T cell clone 1451:1464 a T cell clone 1451:1464 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 9 21 theme glycopeptides 1636:1648 arg1 synthesis 1610:1618 chemo-enzymatic synthesis 1594:1618 chemo-enzymatic synthesis of mannosylated glycopeptides 1594:1648 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 9 22 gly glycopeptides 1636:1648 arg2 glycopeptides 1636:1648 mannosylated glycopeptides 1623:1648 mannosylated glycopeptides 1623:1648 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 4 23 theme homogeneous 849:859 arg1 structures 883:892 defined homogeneous high-mannose N-glycan structures 841:892 defined homogeneous high-mannose N-glycan structures 841:892 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 5 24 theme enhanced 955:962 arg1 targeting 964:972 enhanced targeting 955:972 enhanced targeting 955:972 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 3 25 contain containing 633:642 arg2 N-acetylglucosamine 644:662 N-acetylglucosamine 644:662 N-acetylglucosamine 644:662 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 25 contain containing 633:642 arg1 peptide 511:517 A peptide 509:517 A peptide 509:517 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 25 contain containing 633:642 arg1 substrate 761:769 an acceptor substrate 749:769 an acceptor substrate 749:769 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 4 26 theme N-glycan 874:881 arg1 structures 883:892 defined homogeneous high-mannose N-glycan structures 841:892 defined homogeneous high-mannose N-glycan structures 841:892 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 1 27 theme full-length 338:348 arg1 structures 359:368 full-length N-glycan structures 338:368 full-length N-glycan structures 338:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 7 28 theme N-glycan 1323:1330 arg1 structures 1332:1341 high-mannose N-glycan structures 1310:1341 high-mannose N-glycan structures 1310:1341 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 8 29 theme glycan 1501:1506 arg1 addition 1480:1487 the addition 1476:1487 the addition of a second glycan within the T cell epitope 1476:1532 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 2 30 theme donor 491:495 arg1 substrates 497:506 oxazoline donor substrates 481:506 oxazoline donor substrates 481:506 Mannose-terminated N-glycan oligosaccharides, produced by either total or semi-synthesis, were converted into oxazoline donor substrates. 28717478 0 31 theme mannosylated 40:51 arg1 glycopeptides 53:65 mannosylated glycopeptides 40:65 mannosylated glycopeptides 40:65 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 9 32 theme peptide 1719:1725 arg1 epitopes 1727:1734 peptide epitopes 1719:1734 peptide epitopes 1719:1734 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 3 33 used used 741:744 arg2 substrate 761:769 an acceptor substrate 749:769 an acceptor substrate 749:769 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 33 used used 741:744 arg2 peptide 511:517 A peptide 509:517 A peptide 509:517 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 34 theme protein 565:571 arg1 pp65 573:576 the human cytomegalovirus (CMV) tegument protein pp65 524:576 the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope 524:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 5 35 theme 9-mer 1147:1151 arg1 epitope 1153:1159 a 9-mer epitope 1145:1159 a 9-mer epitope within the peptide 1145:1178 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 3 36 theme acceptor 752:759 arg1 substrate 761:769 an acceptor substrate 749:769 an acceptor substrate 749:769 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 36 theme acceptor 752:759 arg1 peptide 511:517 A peptide 509:517 A peptide 509:517 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 5 37 gly glycopeptides 925:937 arg2 glycopeptides 925:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 0 38 gly glycopeptides 53:65 arg2 glycopeptides 53:65 mannosylated glycopeptides 40:65 mannosylated glycopeptides 40:65 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 9 39 theme mannosylated 1623:1634 arg1 glycopeptides 1636:1648 mannosylated glycopeptides 1623:1648 mannosylated glycopeptides 1623:1648 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 3 40 theme cytomegalovirus 534:548 arg1 pp65 573:576 the human cytomegalovirus (CMV) tegument protein pp65 524:576 the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope 524:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 1 41 theme convergent 269:278 arg1 combination 142:152 The combination 138:152 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 138:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 41 theme convergent 269:278 arg1 method 290:295 a powerful convergent synthetic method 258:295 a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures 258:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 6 42 theme increased 1203:1211 arg1 binding 1213:1219 increased binding 1203:1219 increased binding of glycopeptides to APCs 1203:1244 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 5 43 theme human 1113:1117 arg1 cells 1126:1130 human CD8+ T cells 1113:1130 human CD8+ T cells specific for a 9-mer epitope within the peptide 1113:1178 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 3 44 theme T 617:617 arg1 epitope 624:630 a well-characterised T cell epitope 596:630 a well-characterised T cell epitope 596:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 45 theme human 528:532 arg1 cytomegalovirus 534:548 the human cytomegalovirus 524:548 the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope 524:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 45 theme human 528:532 arg1 CMV 551:553 CMV 551:553 CMV 551:553 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 5 46 theme high-mannose 901:912 arg1 glycopeptides 925:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 2 47 theme N-glycan 390:397 arg1 oligosaccharides 399:414 Mannose-terminated N-glycan oligosaccharides 371:414 Mannose-terminated N-glycan oligosaccharides 371:414 Mannose-terminated N-glycan oligosaccharides, produced by either total or semi-synthesis, were converted into oxazoline donor substrates. 28717478 3 48 from pp65 573:576 arg1 substrate 761:769 an acceptor substrate 749:769 an acceptor substrate 749:769 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 3 48 from pp65 573:576 arg1 peptide 511:517 A peptide 509:517 A peptide 509:517 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 5 49 theme human 977:981 arg1 APCs 1009:1012 APCs 1009:1012 APCs 1009:1012 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 5 49 theme human 977:981 arg1 cells 1002:1006 human antigen presenting cells 977:1006 human antigen presenting cells (APCs) 977:1013 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 3 50 theme specific 667:674 arg1 residues 680:687 specific Asn residues 667:687 specific Asn residues 667:687 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 4 51 gly glycopeptides 819:831 arg2 glycopeptides 819:831 glycopeptides 819:831 glycopeptides bearing defined homogeneous high-mannose N-glycan structures 819:892 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 5 52 theme presenting 991:1000 arg1 APCs 1009:1012 APCs 1009:1012 APCs 1009:1012 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 5 52 theme presenting 991:1000 arg1 cells 1002:1006 human antigen presenting cells 977:1006 human antigen presenting cells (APCs) 977:1013 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 8 53 theme T 1558:1558 arg1 activation 1565:1574 T cell activation 1558:1574 T cell activation 1558:1574 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 3 54 theme Asn 676:678 arg1 residues 680:687 specific Asn residues 667:687 specific Asn residues 667:687 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 1 55 gly glycopeptides 316:328 arg2 glycopeptides 316:328 glycopeptides 316:328 glycopeptides bearing full-length N-glycan structures 316:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 5 56 theme mannose 1044:1050 arg1 receptor 1052:1059 the mannose receptor 1040:1059 the mannose receptor 1040:1059 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 8 57 theme activation 1565:1574 arg1 ablation 1546:1553 ablation 1546:1553 ablation of T cell activation 1546:1574 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 8 58 theme cell 1521:1524 arg1 epitope 1526:1532 the T cell epitope 1515:1532 the T cell epitope 1515:1532 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 3 59 theme solid 706:710 arg1 synthesis 726:734 solid phase peptide synthesis 706:734 solid phase peptide synthesis 706:734 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 9 60 theme chemo-enzymatic 1594:1608 arg1 synthesis 1610:1618 chemo-enzymatic synthesis 1594:1618 chemo-enzymatic synthesis of mannosylated glycopeptides 1594:1648 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 0 61 theme putative 81:88 arg1 candidates 98:107 putative vaccine candidates 81:107 putative vaccine candidates 81:107 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 1 62 theme solid 157:161 arg1 synthesis 177:185 solid phase peptide synthesis 157:185 solid phase peptide synthesis 157:185 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 0 63 theme candidates 98:107 arg1 targeting 68:76 targeting 68:76 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 0:136 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 1 64 theme peptide 169:175 arg1 synthesis 177:185 solid phase peptide synthesis 157:185 solid phase peptide synthesis 157:185 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 6 65 gly glycopeptides 1224:1236 arg2 glycopeptides 1224:1236 glycopeptides 1224:1236 glycopeptides 1224:1236 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 4 66 theme High-yielding 772:784 arg1 glycosylation 796:808 High-yielding enzymatic glycosylation 772:808 High-yielding enzymatic glycosylation 772:808 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 0 67 theme antigen 112:118 arg1 cells 131:135 antigen presenting cells 112:135 antigen presenting cells 112:135 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 5 68 theme CD8+ 1119:1122 arg1 cells 1126:1130 human CD8+ T cells 1113:1130 human CD8+ T cells specific for a 9-mer epitope within the peptide 1113:1178 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 9 69 gly mannosylated 1623:1634 arg1 glycopeptides 1636:1648 mannosylated glycopeptides 1623:1648 mannosylated glycopeptides 1623:1648 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 0 70 theme Convergent 0:9 arg1 synthesis 27:35 Convergent chemo-enzymatic synthesis 0:35 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 0:136 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 1 71 theme catalysed 231:239 arg1 glycosylation 241:253 endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 191:253 endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 191:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 9 72 theme human 1669:1673 arg1 APCs 1675:1678 human APCs 1669:1678 human APCs 1669:1678 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 7 73 theme T 1453:1453 arg1 clone 1460:1464 a T cell clone 1451:1464 a T cell clone 1451:1464 Glycopeptides bearing high-mannose N-glycan structures at a single site outside the T cell epitope were processed and presented by the APCs to allow activation of a T cell clone. 28717478 0 74 theme chemo-enzymatic 11:25 arg1 synthesis 27:35 Convergent chemo-enzymatic synthesis 0:35 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 0:136 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 6 75 theme Binding 1181:1187 arg1 assays 1189:1194 Binding assays 1181:1194 Binding assays 1181:1194 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 4 76 theme defined 841:847 arg1 structures 883:892 defined homogeneous high-mannose N-glycan structures 841:892 defined homogeneous high-mannose N-glycan structures 841:892 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 4 77 theme high-mannose 861:872 arg1 structures 883:892 defined homogeneous high-mannose N-glycan structures 841:892 defined homogeneous high-mannose N-glycan structures 841:892 High-yielding enzymatic glycosylation afforded glycopeptides bearing defined homogeneous high-mannose N-glycan structures. 28717478 5 78 theme containing 914:923 arg1 glycopeptides 925:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides 895:937 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 1 79 theme N-glycan 350:357 arg1 structures 359:368 full-length N-glycan structures 338:368 full-length N-glycan structures 338:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 80 theme glycosylation 241:253 arg1 combination 142:152 The combination 138:152 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 138:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 80 theme glycosylation 241:253 arg1 method 290:295 a powerful convergent synthetic method 258:295 a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures 258:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 5 81 theme specific 1132:1139 arg1 cells 1126:1130 human CD8+ T cells 1113:1130 human CD8+ T cells specific for a 9-mer epitope within the peptide 1113:1178 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 3 82 theme peptide 718:724 arg1 synthesis 726:734 solid phase peptide synthesis 706:734 solid phase peptide synthesis 706:734 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 8 83 theme second 1494:1499 arg1 glycan 1501:1506 a second glycan 1492:1506 a second glycan 1492:1506 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 2 84 theme oxazoline 481:489 arg1 substrates 497:506 oxazoline donor substrates 481:506 oxazoline donor substrates 481:506 Mannose-terminated N-glycan oligosaccharides, produced by either total or semi-synthesis, were converted into oxazoline donor substrates. 28717478 9 85 theme epitopes 1727:1734 arg1 immunogenicity 1701:1714 the immunogenicity 1697:1714 the immunogenicity of peptide epitopes 1697:1734 We conclude that chemo-enzymatic synthesis of mannosylated glycopeptides enhances uptake by human APCs while preserving the immunogenicity of peptide epitopes within the glycopeptides, provided those epitopes are not themselves glycosylated. 28717478 0 86 gly mannosylated 40:51 arg1 glycopeptides 53:65 mannosylated glycopeptides 40:65 mannosylated glycopeptides 40:65 Convergent chemo-enzymatic synthesis of mannosylated glycopeptides; targeting of putative vaccine candidates to antigen presenting cells. 28717478 8 87 theme T 1519:1519 arg1 epitope 1526:1532 the T cell epitope 1515:1532 the T cell epitope 1515:1532 However, the addition of a second glycan within the T cell epitope resulted in ablation of T cell activation. 28717478 3 88 theme tegument 556:563 arg1 pp65 573:576 the human cytomegalovirus (CMV) tegument protein pp65 524:576 the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope 524:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 1 89 theme powerful 260:267 arg1 combination 142:152 The combination 138:152 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 138:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 89 theme powerful 260:267 arg1 method 290:295 a powerful convergent synthetic method 258:295 a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures 258:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 5 90 theme T 1124:1124 arg1 cells 1126:1130 human CD8+ T cells 1113:1130 human CD8+ T cells specific for a 9-mer epitope within the peptide 1113:1178 These high-mannose containing glycopeptides were tested for enhanced targeting to human antigen presenting cells (APCs), putatively mediated via the mannose receptor, and for processing by the APCs for presentation to human CD8+ T cells specific for a 9-mer epitope within the peptide. 28717478 6 91 theme glycopeptides 1224:1236 arg1 binding 1213:1219 increased binding 1203:1219 increased binding of glycopeptides to APCs 1203:1244 Binding assays showed increased binding of glycopeptides to APCs compared to the non-glycosylated control. 28717478 3 92 theme well-characterised 598:615 arg1 epitope 624:630 a well-characterised T cell epitope 596:630 a well-characterised T cell epitope 596:630 A peptide from the human cytomegalovirus (CMV) tegument protein pp65 that incorporates a well-characterised T cell epitope, containing N-acetylglucosamine at specific Asn residues, was accessed by solid phase peptide synthesis, and used as an acceptor substrate. 28717478 1 93 theme synthetic 280:288 arg1 combination 142:152 The combination 138:152 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation 138:253 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 28717478 1 93 theme synthetic 280:288 arg1 method 290:295 a powerful convergent synthetic method 258:295 a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures 258:368 The combination of solid phase peptide synthesis and endo-β-N-acetylglucosaminidase (ENGase) catalysed glycosylation is a powerful convergent synthetic method allowing access to glycopeptides bearing full-length N-glycan structures. 27769287 0 0 theme recombinant 53:63 arg1 glycoproteins 65:77 recombinant glycoproteins 53:77 recombinant glycoproteins 53:77 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 1 1 theme human 326:330 arg1 sugars 332:337 human sugars 326:337 human sugars 326:337 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 4 2 theme enhanced 847:854 arg1 layer 863:867 an enhanced glucan layer 844:867 an enhanced glucan layer 844:867 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 5 3 theme catalytic 1088:1096 arg1 protein 1077:1083 Fks2 protein 1072:1083 Fks2 protein 1072:1083 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 5 3 theme catalytic 1088:1096 arg1 subunit 1098:1104 a catalytic subunit 1086:1104 a catalytic subunit of a 1,3-β-glucan synthase 1086:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 1 4 theme N-glycosylation 138:152 arg1 pathways 154:161 yeast N-glycosylation pathways 132:161 yeast N-glycosylation pathways 132:161 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 6 5 located observed 1332:1339 arg1 cells 1364:1368 the Δalg3Δoch1-FKS2 cells 1344:1368 the Δalg3Δoch1-FKS2 cells 1344:1368 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 6 5 located observed 1332:1339 arg2 abilities 1241:1249 improved abilities 1232:1249 improved abilities 1232:1249 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 6 6 from abilities 1241:1249 arg1 sensitivity 1304:1314 drug sensitivity 1299:1314 drug sensitivity 1299:1314 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 6 6 from abilities 1241:1249 arg1 tolerance 1266:1274 temperature tolerance 1254:1274 temperature tolerance 1254:1274 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 6 6 from abilities 1241:1249 arg1 potential 1285:1293 osmotic potential 1277:1293 osmotic potential 1277:1293 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 4 7 theme cell 950:953 arg1 biosynthesis 960:971 cell wall biosynthesis 950:971 cell wall biosynthesis 950:971 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 1 8 theme sugars 332:337 arg1 synthesis 288:296 synthesis 288:296 synthesis 288:296 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 1 8 theme sugars 332:337 arg1 transport 299:307 transport 299:307 transport 299:307 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 1 8 theme sugars 332:337 arg1 addition 314:321 addition 314:321 addition 314:321 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 6 9 theme osmotic 1277:1283 arg1 potential 1285:1293 osmotic potential 1277:1293 osmotic potential 1277:1293 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 2 10 theme cell 461:464 arg1 growth 466:471 cell growth 461:471 cell growth 461:471 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 3 11 theme small 602:606 arg1 GTPase 608:613 the Rho1p small GTPase 592:613 the Rho1p small GTPase 592:613 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 3 11 theme small 602:606 arg1 overexpression 554:567 overexpression 554:567 overexpression 554:567 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 8 12 theme RHO1 1518:1521 arg1 Overexpression 1500:1513 Overexpression 1500:1513 Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts 1500:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 8 13 from Overexpression 1500:1513 arg1 yeasts 1561:1566 'humanized' glycoprotein producing yeasts 1526:1566 'humanized' glycoprotein producing yeasts 1526:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 0 14 from Effects 0:6 arg1 production 39:48 the production 35:48 the production of recombinant glycoproteins in Saccharomyces cerevisiae 35:105 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 6 15 theme wall 1163:1166 arg1 structure 1168:1176 cell wall structure 1158:1176 cell wall structure 1158:1176 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 4 16 from effect 924:929 arg1 biosynthesis 960:971 cell wall biosynthesis 950:971 cell wall biosynthesis 950:971 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 0 17 theme glycoproteins 65:77 arg1 production 39:48 the production 35:48 the production of recombinant glycoproteins in Saccharomyces cerevisiae 35:105 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 8 18 theme humanized 1527:1535 arg1 yeasts 1561:1566 'humanized' glycoprotein producing yeasts 1526:1566 'humanized' glycoprotein producing yeasts 1526:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 4 19 theme glucan 856:861 arg1 layer 863:867 an enhanced glucan layer 844:867 an enhanced glucan layer 844:867 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 8 20 theme glycoproteins 1668:1680 arg1 production 1642:1651 the production 1638:1651 the production of therapeutic glycoproteins 1638:1680 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 6 21 theme cell 1158:1161 arg1 structure 1168:1176 cell wall structure 1158:1176 cell wall structure 1158:1176 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 1 22 theme genes 267:271 arg1 introduction 251:262 the introduction 247:262 the introduction of genes catalyzing the synthesis, transport, and addition of human sugars 247:337 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 4 23 theme Rho1p 934:938 arg1 GTPase 940:945 Rho1p GTPase 934:945 Rho1p GTPase 934:945 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 3 24 theme Δalg3Δoch1 696:705 arg1 strain 721:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 4 25 theme RHO1-expressed 818:831 arg1 cells 833:837 RHO1-expressed cells 818:837 RHO1-expressed cells 818:837 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 4 26 theme transmission 742:753 arg1 micrographs 764:774 transmission electron micrographs 742:774 transmission electron micrographs 742:774 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 5 27 theme Fks2 1072:1075 arg1 protein 1077:1083 Fks2 protein 1072:1083 Fks2 protein 1072:1083 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 5 27 theme Fks2 1072:1075 arg1 gene 1054:1057 the gene 1050:1057 the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase 1050:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 5 27 theme Fks2 1072:1075 arg1 subunit 1098:1104 a catalytic subunit 1086:1104 a catalytic subunit of a 1,3-β-glucan synthase 1086:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 0 28 theme Saccharomyces 82:94 arg1 cerevisiae 96:105 Saccharomyces cerevisiae 82:105 Saccharomyces cerevisiae 82:105 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 2 29 from defects 450:456 arg1 growth 466:471 cell growth 461:471 cell growth 461:471 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 2 29 from defects 450:456 arg1 response 492:499 response 492:499 response to environmental challenges 492:527 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 2 29 from defects 450:456 arg1 morphogenesis 474:486 morphogenesis 474:486 morphogenesis 474:486 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 8 30 theme therapeutic 1656:1666 arg1 glycoproteins 1668:1680 therapeutic glycoproteins 1656:1680 therapeutic glycoproteins 1656:1680 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 7 31 theme site 1430:1433 arg1 occupancy 1435:1443 N-glycan site occupancy 1421:1443 N-glycan site occupancy 1421:1443 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 8 32 theme producing 1551:1559 arg1 yeasts 1561:1566 'humanized' glycoprotein producing yeasts 1526:1566 'humanized' glycoprotein producing yeasts 1526:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 7 33 theme RHO1 1381:1384 arg1 overexpression 1386:1399 RHO1 overexpression 1381:1399 RHO1 overexpression 1381:1399 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 3 34 theme Saccharomyces 671:683 arg1 strain 721:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 0 35 gly glycoproteins 65:77 arg1 glycoproteins 65:77 recombinant glycoproteins 53:77 recombinant glycoproteins 53:77 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 7 36 theme secreted 1463:1470 arg1 glycoproteins 1472:1484 secreted glycoproteins 1463:1484 secreted glycoproteins 1463:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 2 37 theme genes 367:371 arg1 deletion 349:356 deletion 349:356 deletion of these genes 349:371 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 7 38 theme glycoproteins 1472:1484 arg1 occupancy 1435:1443 N-glycan site occupancy 1421:1443 N-glycan site occupancy 1421:1443 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 7 38 theme glycoproteins 1472:1484 arg1 amount 1453:1458 the amount 1449:1458 the amount of secreted glycoproteins 1449:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 7 38 theme glycoproteins 1472:1484 arg1 glycoproteins 1472:1484 secreted glycoproteins 1463:1484 secreted glycoproteins 1463:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 3 39 theme strain 721:726 arg1 defect 661:666 the growth defect 650:666 the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 650:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 4 40 theme cells 833:837 arg1 structure 805:813 the cell wall structure 791:813 the cell wall structure of RHO1-expressed cells 791:837 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 6 41 theme temperature 1254:1264 arg1 tolerance 1266:1274 temperature tolerance 1254:1274 temperature tolerance 1254:1274 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 8 42 gly glycoproteins 1668:1680 arg1 glycoproteins 1668:1680 therapeutic glycoproteins 1656:1680 therapeutic glycoproteins 1656:1680 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 6 43 theme Δalg3Δoch1 1202:1211 arg1 strain 1213:1218 the RHO1-overexpressed Δalg3Δoch1 strain 1179:1218 the RHO1-overexpressed Δalg3Δoch1 strain 1179:1218 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 4 44 theme wall 955:958 arg1 biosynthesis 960:971 cell wall biosynthesis 950:971 cell wall biosynthesis 950:971 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 1 45 theme yeast 182:186 arg1 hyper-mannosylation 197:215 yeast specific hyper-mannosylation 182:215 yeast specific hyper-mannosylation 182:215 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 0 46 theme Rho1 11:14 arg1 Effects 0:6 Effects 0:6 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae 0:105 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 3 47 theme double 707:712 arg1 strain 721:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 4 48 theme mannoprotein 890:901 arg1 layer 903:907 a recovered mannoprotein layer 878:907 a recovered mannoprotein layer 878:907 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 5 49 theme gene 1054:1057 arg1 overexpression 1032:1045 overexpression 1032:1045 overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase 1032:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 8 50 gly glycoprotein 1538:1549 arg1 glycoprotein 1538:1549 'humanized' glycoprotein producing yeasts 1526:1566 'humanized' glycoprotein producing yeasts 1526:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 8 51 theme glycoprotein 1538:1549 arg1 yeasts 1561:1566 'humanized' glycoprotein producing yeasts 1526:1566 'humanized' glycoprotein producing yeasts 1526:1566 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 1 52 theme specific 188:195 arg1 hyper-mannosylation 197:215 yeast specific hyper-mannosylation 182:215 yeast specific hyper-mannosylation 182:215 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 0 53 theme small 19:23 arg1 Rho1 11:14 Rho1 11:14 Rho1 11:14 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 0 53 theme small 19:23 arg1 GTPase 25:30 a small GTPase 17:30 a small GTPase 17:30 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 3 54 theme mutant 714:719 arg1 strain 721:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 671:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 4 55 theme cell 795:798 arg1 structure 805:813 the cell wall structure 791:813 the cell wall structure of RHO1-expressed cells 791:837 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 7 56 gly glycoproteins 1472:1484 arg1 glycoproteins 1472:1484 secreted glycoproteins 1463:1484 secreted glycoproteins 1463:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 2 57 theme severe 443:448 arg1 defects 450:456 severe defects 443:456 severe defects in cell growth, morphogenesis and response to environmental challenges 443:527 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 5 58 theme synthase 1124:1131 arg1 protein 1077:1083 Fks2 protein 1072:1083 Fks2 protein 1072:1083 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 5 58 theme synthase 1124:1131 arg1 subunit 1098:1104 a catalytic subunit 1086:1104 a catalytic subunit of a 1,3-β-glucan synthase 1086:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 2 59 theme environmental 504:516 arg1 challenges 518:527 environmental challenges 504:527 environmental challenges 504:527 However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. 27769287 0 60 from production 39:48 arg1 cerevisiae 96:105 Saccharomyces cerevisiae 82:105 Saccharomyces cerevisiae 82:105 Effects of Rho1, a small GTPase on the production of recombinant glycoproteins in Saccharomyces cerevisiae. 27769287 3 61 theme RHO1 572:575 arg1 GTPase 608:613 the Rho1p small GTPase 592:613 the Rho1p small GTPase 592:613 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 3 61 theme RHO1 572:575 arg1 overexpression 554:567 overexpression 554:567 overexpression 554:567 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 8 62 theme future 1603:1608 arg1 applications 1621:1632 its future industrial applications 1599:1632 its future industrial applications for the production of therapeutic glycoproteins 1599:1680 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 3 63 dep Saccharomyces 671:683 arg1 cerevisiae 685:694 cerevisiae 685:694 cerevisiae 685:694 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 1 64 dep disrupted 225:233 arg1 followed 235:242 followed 235:242 followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars 235:337 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 6 65 theme drug 1299:1302 arg1 sensitivity 1304:1314 drug sensitivity 1299:1314 drug sensitivity 1299:1314 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 6 66 theme improved 1232:1239 arg1 abilities 1241:1249 improved abilities 1232:1249 improved abilities 1232:1249 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 5 67 theme 1,3-β-glucan 1111:1122 arg1 synthase 1124:1131 a 1,3-β-glucan synthase 1109:1131 a 1,3-β-glucan synthase 1109:1131 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 8 68 theme industrial 1610:1619 arg1 applications 1621:1632 its future industrial applications 1599:1632 its future industrial applications for the production of therapeutic glycoproteins 1599:1680 CONCLUSIONS: Overexpression of RHO1 in 'humanized' glycoprotein producing yeasts could significantly facilitate its future industrial applications for the production of therapeutic glycoproteins. 27769287 3 69 theme growth 654:659 arg1 defect 661:666 the growth defect 650:666 the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain 650:726 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 4 70 theme wall 800:803 arg1 structure 805:813 the cell wall structure 791:813 the cell wall structure of RHO1-expressed cells 791:837 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 3 71 theme Rho1p 596:600 arg1 GTPase 608:613 the Rho1p small GTPase 592:613 the Rho1p small GTPase 592:613 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 3 71 theme Rho1p 596:600 arg1 overexpression 554:567 overexpression 554:567 overexpression 554:567 RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain. 27769287 6 72 theme structure 1168:1176 arg1 recovery 1146:1153 the recovery 1142:1153 the recovery of cell wall structure 1142:1176 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 4 73 theme recovered 880:888 arg1 layer 903:907 a recovered mannoprotein layer 878:907 a recovered mannoprotein layer 878:907 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 6 74 theme RHO1-overexpressed 1183:1200 arg1 strain 1213:1218 the RHO1-overexpressed Δalg3Δoch1 strain 1179:1218 the RHO1-overexpressed Δalg3Δoch1 strain 1179:1218 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 27769287 7 75 theme N-glycan 1421:1428 arg1 occupancy 1435:1443 N-glycan site occupancy 1421:1443 N-glycan site occupancy 1421:1443 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 4 76 contain have 839:842 arg2 layer 863:867 an enhanced glucan layer 844:867 an enhanced glucan layer 844:867 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 4 76 contain have 839:842 arg2 layer 903:907 a recovered mannoprotein layer 878:907 a recovered mannoprotein layer 878:907 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 4 76 contain have 839:842 arg1 structure 805:813 the cell wall structure 791:813 the cell wall structure of RHO1-expressed cells 791:837 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 4 77 theme electron 755:762 arg1 micrographs 764:774 transmission electron micrographs 742:774 transmission electron micrographs 742:774 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 5 78 theme Similar 974:980 arg1 phenotypes 998:1007 Similar complementation phenotypes 974:1007 Similar complementation phenotypes 974:1007 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 7 79 gly occupancy 1435:1443 arg2 site 1430:1433 N-glycan site occupancy 1421:1443 N-glycan site occupancy 1421:1443 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 7 79 gly occupancy 1435:1443 arg2 glycoproteins 1472:1484 secreted glycoproteins 1463:1484 secreted glycoproteins 1463:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 7 79 gly occupancy 1435:1443 arg1 glycoproteins 1472:1484 secreted glycoproteins 1463:1484 secreted glycoproteins 1463:1484 Moreover, RHO1 overexpression could also increase N-glycan site occupancy and the amount of secreted glycoproteins. 27769287 4 80 theme GTPase 940:945 arg1 effect 924:929 the effect 920:929 the effect of Rho1p GTPase on cell wall biosynthesis 920:971 In addition, transmission electron micrographs indicated that the cell wall structure of RHO1-expressed cells have an enhanced glucan layer and also a recovered mannoprotein layer, revealing the effect of Rho1p GTPase on cell wall biosynthesis. 27769287 1 81 dep synthesis 288:296 arg1 the 284:286 the 284:286 the 284:286 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 5 82 theme complementation 982:996 arg1 phenotypes 998:1007 Similar complementation phenotypes 974:1007 Similar complementation phenotypes 974:1007 Similar complementation phenotypes have been confirmed by overexpression of the gene that encodes Fks2 protein, a catalytic subunit of a 1,3-β-glucan synthase. 27769287 1 83 theme yeast 132:136 arg1 pathways 154:161 yeast N-glycosylation pathways 132:161 yeast N-glycosylation pathways 132:161 BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. 27769287 6 84 theme Δalg3Δoch1-FKS2 1348:1362 arg1 cells 1364:1368 the Δalg3Δoch1-FKS2 cells 1344:1368 the Δalg3Δoch1-FKS2 cells 1344:1368 Besides the recovery of cell wall structure, the RHO1-overexpressed Δalg3Δoch1 strain also showed improved abilities in temperature tolerance, osmotic potential and drug sensitivity, which were not observed in the Δalg3Δoch1-FKS2 cells. 24201825 0 0 theme cascade 76:82 arg1 proteases 84:92 coagulation cascade proteases 64:92 coagulation cascade proteases 64:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 3 1 theme differential 670:681 arg1 inhibition 683:692 this differential inhibition 665:692 this differential inhibition 665:692 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 6 2 theme ternary 1302:1308 arg1 complexes 1310:1318 ternary complexes 1302:1318 ternary complexes 1302:1318 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 7 3 theme strong 1394:1399 arg1 glycosaminoglycan 1401:1417 a strong glycosaminoglycan 1392:1417 a strong glycosaminoglycan binding to the proteases exosite 2 1392:1452 Finally, the enzymes catalytic triad organization was disrupted due to a strong glycosaminoglycan binding to the proteases exosite 2. 24201825 0 4 theme coagulation 64:74 arg1 proteases 84:92 coagulation cascade proteases 64:92 coagulation cascade proteases 64:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 0 5 from glycobiology 11:22 arg1 proteases 84:92 coagulation cascade proteases 64:92 coagulation cascade proteases 64:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 0 5 from glycobiology 11:22 arg1 exosite 51:57 exosite 2 51:59 the exosite 2 of coagulation cascade proteases 47:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 2 6 theme complexes 329:337 arg1 formation 308:316 The formation 304:316 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, 304:396 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 2 6 theme complexes 329:337 arg1 path 410:413 the main path 401:413 the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action 401:501 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 2 7 theme action 496:501 arg1 mechanisms 482:491 two distinct mechanisms 469:491 two distinct mechanisms of action 469:501 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 4 8 theme glycosylation 853:865 arg1 effects 842:848 the effects 838:848 the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics 838:953 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 6 9 theme heparin 1174:1180 arg1 orientation 1182:1192 heparin orientation 1174:1192 heparin orientation observed on the surface of fIIa, but not fXa, 1174:1238 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 8 10 theme inhibition 1516:1525 arg1 constant 1527:1534 the higher inhibition constant 1505:1534 the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa 1505:1589 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 0 11 theme proteases 84:92 arg1 proteases 84:92 coagulation cascade proteases 64:92 coagulation cascade proteases 64:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 0 11 theme proteases 84:92 arg1 exosite 51:57 exosite 2 51:59 the exosite 2 of coagulation cascade proteases 47:92 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 6 12 from antithrombin 1286:1297 arg1 complexes 1310:1318 ternary complexes 1302:1318 ternary complexes 1302:1318 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 6 13 theme fIIa 1221:1224 arg1 surface 1210:1216 the surface 1206:1216 the surface of fIIa, but not fXa, 1206:1238 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 3 14 theme serine 516:521 arg1 proteases 523:531 these serine proteases 510:531 these serine proteases 510:531 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 6 15 theme long-chain 1256:1265 arg1 heparin 1267:1273 a linear long-chain heparin 1247:1273 a linear long-chain heparin binding to antithrombin in ternary complexes 1247:1318 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 1 16 theme major 256:260 arg1 role 262:265 a major role 254:265 a major role 254:265 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 4 17 theme molecular 807:815 arg1 simulations 826:836 molecular dynamics simulations 807:836 molecular dynamics simulations 807:836 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 8 18 theme different 1611:1619 arg1 susceptibilities 1621:1636 the different susceptibilities 1607:1636 the different susceptibilities of those enzymes for antithrombin mechanisms of action 1607:1691 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 8 19 theme enzymes 1647:1653 arg1 susceptibilities 1621:1636 the different susceptibilities 1607:1636 the different susceptibilities of those enzymes for antithrombin mechanisms of action 1607:1691 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 2 20 theme distinct 473:480 arg1 mechanisms 482:491 two distinct mechanisms 469:491 two distinct mechanisms of action 469:501 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 3 21 theme inhibition 683:692 arg1 basis 656:660 the molecular basis 642:660 the molecular basis of this differential inhibition 642:692 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 8 22 theme antithrombin-heparin 1543:1562 arg1 complex 1564:1570 the antithrombin-heparin complex 1539:1570 the antithrombin-heparin complex over fIIa 1539:1580 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 7 23 theme catalytic 1342:1350 arg1 organization 1358:1369 the enzymes catalytic triad organization 1330:1369 the enzymes catalytic triad organization 1330:1369 Finally, the enzymes catalytic triad organization was disrupted due to a strong glycosaminoglycan binding to the proteases exosite 2. 24201825 7 24 dep proteases 1434:1442 arg1 exosite 1444:1450 exosite 2 1444:1452 the proteases exosite 2 1430:1452 Finally, the enzymes catalytic triad organization was disrupted due to a strong glycosaminoglycan binding to the proteases exosite 2. 24201825 3 25 theme distinct 541:548 arg1 susceptibilities 550:565 distinct susceptibilities 541:565 distinct susceptibilities 541:565 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 8 26 theme antithrombin 1659:1670 arg1 mechanisms 1672:1681 antithrombin mechanisms 1659:1681 antithrombin mechanisms of action 1659:1691 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 0 27 theme Structural 0:9 arg1 glycobiology 11:22 Structural glycobiology 0:22 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 0:154 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 3 28 theme glycosylation 715:727 arg1 role 702:705 the role 698:705 the role of fIIa glycosylation on this process 698:743 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 4 29 theme dynamics 817:824 arg1 simulations 826:836 molecular dynamics simulations 807:836 molecular dynamics simulations 807:836 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 7 30 theme triad 1352:1356 arg1 organization 1358:1369 the enzymes catalytic triad organization 1330:1369 the enzymes catalytic triad organization 1330:1369 Finally, the enzymes catalytic triad organization was disrupted due to a strong glycosaminoglycan binding to the proteases exosite 2. 24201825 8 31 theme action 1686:1691 arg1 mechanisms 1672:1681 antithrombin mechanisms 1659:1681 antithrombin mechanisms of action 1659:1691 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 1 32 theme main 184:187 arg1 targets 189:195 the main targets 180:195 the main targets of antithrombin 180:211 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 0 33 theme heparin 27:33 arg1 dynamics 35:42 heparin dynamics 27:42 heparin dynamics 27:42 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 5 34 theme obtained 969:976 arg1 data 978:981 the obtained data 965:981 the obtained data 965:981 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 0 35 theme antithrombotic 131:144 arg1 activity 146:153 glycosaminoglycans antithrombotic activity 112:153 glycosaminoglycans antithrombotic activity 112:153 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 3 36 with interaction 608:618 arg1 heparin 625:631 heparin 625:631 heparin 625:631 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 1 37 theme targets 189:195 arg1 two 173:175 two 173:175 two 173:175 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 1 37 theme targets 189:195 arg1 inhibitor 233:241 a serine proteases inhibitor 214:241 a serine proteases inhibitor that plays a major role in the regulation of blood clotting 214:301 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 1 37 theme targets 189:195 arg1 targets 189:195 the main targets 180:195 the main targets of antithrombin 180:211 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 2 38 theme main 405:408 arg1 path 410:413 the main path 401:413 the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action 401:501 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 2 38 theme main 405:408 arg1 formation 308:316 The formation 304:316 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, 304:396 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 8 39 theme higher 1509:1514 arg1 constant 1527:1534 the higher inhibition constant 1505:1534 the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa 1505:1589 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 0 40 theme glycosaminoglycans 112:129 arg1 activity 146:153 glycosaminoglycans antithrombotic activity 112:153 glycosaminoglycans antithrombotic activity 112:153 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 7 41 theme enzymes 1334:1340 arg1 organization 1358:1369 the enzymes catalytic triad organization 1330:1369 the enzymes catalytic triad organization 1330:1369 Finally, the enzymes catalytic triad organization was disrupted due to a strong glycosaminoglycan binding to the proteases exosite 2. 24201825 5 42 link N-linked 989:996 arg1 glycan 998:1003 fIIa N-linked glycan 984:1003 fIIa N-linked glycan 984:1003 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 3 43 theme fIIa 710:713 arg1 glycosylation 715:727 fIIa glycosylation 710:727 fIIa glycosylation 710:727 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 4 44 theme binding 907:913 arg1 fIIa 870:873 fIIa 870:873 fIIa 870:873 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 4 44 theme binding 907:913 arg1 consequences 883:894 the consequences 879:894 the consequences of heparin binding to both proteases function and dynamics 879:953 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 5 45 theme active 1033:1038 arg1 size 1052:1055 the active site pocket size 1029:1055 the active site pocket size 1029:1055 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 4 46 theme present 776:782 arg1 work 784:787 the present work 772:787 the present work 772:787 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 4 47 gly glycosylation 853:865 arg1 binding 907:913 heparin binding 899:913 heparin binding to both proteases function and dynamics 899:953 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 1 48 theme antithrombin 200:211 arg1 targets 189:195 the main targets 180:195 the main targets of antithrombin 180:211 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 0 49 theme dynamics 35:42 arg1 glycobiology 11:22 Structural glycobiology 0:22 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 0:154 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 4 50 theme heparin 899:905 arg1 binding 907:913 heparin binding 899:913 heparin binding to both proteases function and dynamics 899:953 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 5 51 theme site 1040:1043 arg1 size 1052:1055 the active site pocket size 1029:1055 the active site pocket size 1029:1055 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 5 52 theme fIIa 984:987 arg1 glycan 998:1003 fIIa N-linked glycan 984:1003 fIIa N-linked glycan 984:1003 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 1 53 theme blood 288:292 arg1 clotting 294:301 blood clotting 288:301 blood clotting 288:301 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 4 54 dep proteases 923:931 arg1 function 933:940 function 933:940 function 933:940 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 5 55 theme pocket 1045:1050 arg1 size 1052:1055 the active site pocket size 1029:1055 the active site pocket size 1029:1055 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 5 56 theme N-linked 989:996 arg1 glycan 998:1003 fIIa N-linked glycan 984:1003 fIIa N-linked glycan 984:1003 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 8 57 theme atomic-level 1476:1487 arg1 explanation 1489:1499 an atomic-level explanation 1473:1499 an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action 1473:1691 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 2 58 theme ternary 321:327 arg1 complexes 329:337 ternary complexes 321:337 ternary complexes 321:337 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 6 59 theme linear 1249:1254 arg1 heparin 1267:1273 a linear long-chain heparin 1247:1273 a linear long-chain heparin binding to antithrombin in ternary complexes 1247:1318 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 1 60 theme clotting 294:301 arg1 regulation 274:283 the regulation 270:283 the regulation of blood clotting 270:301 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 6 61 located observed 1194:1201 arg1 surface 1210:1216 the surface 1206:1216 the surface of fIIa, but not fXa, 1206:1238 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 6 61 located observed 1194:1201 arg2 orientation 1182:1192 heparin orientation 1174:1192 heparin orientation observed on the surface of fIIa, but not fXa, 1174:1238 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 3 62 theme molecular 646:654 arg1 basis 656:660 the molecular basis 642:660 the molecular basis of this differential inhibition 642:692 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 8 63 theme complex 1564:1570 arg1 constant 1527:1534 the higher inhibition constant 1505:1534 the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa 1505:1589 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 6 64 theme fXa 1235:1237 arg1 surface 1210:1216 the surface 1206:1216 the surface of fIIa, but not fXa, 1206:1238 Additionally, heparin orientation observed on the surface of fIIa, but not fXa, allows a linear long-chain heparin binding to antithrombin in ternary complexes. 24201825 4 65 from effects 842:848 arg1 fIIa 870:873 fIIa 870:873 fIIa 870:873 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 4 65 from effects 842:848 arg1 consequences 883:894 the consequences 879:894 the consequences of heparin binding to both proteases function and dynamics 879:953 Thus, the present work evaluated through molecular dynamics simulations the effects of glycosylation on fIIa and the consequences of heparin binding to both proteases function and dynamics. 24201825 1 66 theme serine 216:221 arg1 proteases 223:231 serine proteases 216:231 a serine proteases inhibitor that plays a major role in the regulation of blood clotting 214:301 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 5 67 from increase 1017:1024 arg1 size 1052:1055 the active site pocket size 1029:1055 the active site pocket size 1029:1055 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 24201825 0 68 dep glycobiology 11:22 arg1 Implications 95:106 Implications 95:106 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 0:154 Structural glycobiology of heparin dynamics on the exosite 2 of coagulation cascade proteases: Implications for glycosaminoglycans antithrombotic activity. 24201825 1 69 theme proteases 223:231 arg1 two 173:175 two 173:175 two 173:175 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 1 69 theme proteases 223:231 arg1 inhibitor 233:241 a serine proteases inhibitor 214:241 a serine proteases inhibitor that plays a major role in the regulation of blood clotting 214:301 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 1 69 theme proteases 223:231 arg1 targets 189:195 the main targets 180:195 the main targets of antithrombin 180:211 fIIa and fXa are two of the main targets of antithrombin, a serine proteases inhibitor that plays a major role in the regulation of blood clotting. 24201825 3 70 from role 702:705 arg1 process 737:743 this process 732:743 this process 732:743 While these serine proteases present distinct susceptibilities to these paths, in which fIIa demands an interaction with heparin, neither the molecular basis of this differential inhibition nor the role of fIIa glycosylation on this process is fully understood. 24201825 2 71 theme such 347:350 arg1 molecules 352:360 such molecules 347:360 such molecules 347:360 The formation of ternary complexes between such molecules and glycosaminoglycans, as heparin, is the main path for inhibiting those enzymes, which may occur through two distinct mechanisms of action. 24201825 8 72 theme Such 1455:1458 arg1 data 1460:1463 Such data 1455:1463 Such data 1455:1463 Such data support an atomic-level explanation for the higher inhibition constant of the antithrombin-heparin complex over fIIa than fXa, as well as for the different susceptibilities of those enzymes for antithrombin mechanisms of action. 24201825 5 73 theme heparin 1104:1110 arg1 binding 1112:1118 heparin binding 1104:1118 heparin binding 1104:1118 Based on the obtained data, fIIa N-linked glycan promoted an increase in the active site pocket size by stabilizing regions that encircle it, while heparin binding was observed to reverse such an effect. 27506354 9 0 theme 31 N-glycan 1738:1748 arg1 masses 1750:1755 31 N-glycan masses 1738:1755 31 N-glycan masses 1738:1755 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 1 1 theme processes 260:268 arg1 variety 240:246 a wide variety 233:246 a wide variety of cellular processes 233:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 1 1 theme processes 260:268 arg1 processes 260:268 cellular processes 251:268 cellular processes 251:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 1 2 gly glycosylation 190:202 arg1 regulation 219:228 regulation 219:228 regulation of a wide variety of cellular processes 219:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 3 3 from lack 495:498 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 9 4 from profiling 1679:1687 arg1 serum 1711:1715 human serum 1705:1715 human serum 1705:1715 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 0 5 with combination 101:111 arg1 enrichment 129:138 Ti(4+)-SPE enrichment 118:138 Ti(4+)-SPE enrichment for mass spectrometric analysis 118:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 1 6 theme protein 182:188 arg1 glycosylation 190:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 6 7 theme phospho-containing 1106:1123 arg1 glycans 1125:1131 the phospho-containing glycans 1102:1131 the phospho-containing glycans 1102:1131 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 8 8 theme excellent 1405:1413 arg1 selectivity 1415:1425 excellent selectivity 1405:1425 excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100) 1405:1505 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 3 9 theme detection 621:629 arg1 sensitivity 631:641 the particularly poor detection sensitivity 599:641 the particularly poor detection sensitivity of N-glycans 599:654 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 4 10 theme crucial 740:746 arg1 step 748:751 a crucial step 738:751 a crucial step prior to MS analysis of the N-glycome 738:789 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 4 11 theme prior 753:757 arg1 step 748:751 a crucial step 738:751 a crucial step prior to MS analysis of the N-glycome 738:789 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 8 12 theme high 1359:1362 arg1 % 1401:1401 almost 100% 1391:1401 almost 100% 1391:1401 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 12 theme high 1359:1362 arg1 efficiency 1379:1388 high derivatization efficiency 1359:1388 high derivatization efficiency (almost 100%) 1359:1402 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 0 13 theme mass 144:147 arg1 analysis 163:170 mass spectrometric analysis 144:170 mass spectrometric analysis 144:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 4 14 theme purification 707:718 arg1 procedure 720:728 the highly specific purification procedure 687:728 the highly specific purification procedure 687:728 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 7 15 theme maltoheptaose 1214:1226 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 15 theme maltoheptaose 1214:1226 arg1 DP7 1228:1230 maltoheptaose DP7 1214:1230 maltoheptaose DP7 1214:1230 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 9 16 theme proposed 1631:1638 arg1 method 1640:1645 the proposed method 1627:1645 the proposed method 1627:1645 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 0 17 theme affinity 85:92 arg1 tag 94:96 an affinity tag 82:96 an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis 82:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 0 17 theme affinity 85:92 arg1 derivatization 64:77 phosphate-based derivatization 48:77 phosphate-based derivatization 48:77 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 5 18 theme phase 904:908 arg1 extraction 910:919 solid phase extraction 898:919 Ti(4+)-SPE (solid phase extraction) 886:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 8 19 theme serum 1467:1471 arg1 albumin 1473:1479 bovine serum albumin 1460:1479 bovine serum albumin 1460:1479 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 4 20 theme N-glycome 781:789 arg1 analysis 765:772 MS analysis 762:772 MS analysis of the N-glycome 762:789 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 1 21 link N-linked 173:180 arg1 glycosylation 190:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 6 22 dep such 1043:1046 arg1 able 1086:1089 able 1086:1089 able 1086:1089 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 6 22 dep such 1043:1046 arg1 microspheres 1068:1079 the Ti(4+)-SPE microspheres 1053:1079 the Ti(4+)-SPE microspheres 1053:1079 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 3 23 theme glycan 456:461 arg1 abundance 443:451 The relatively low abundance 424:451 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides 424:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 8 24 from DP7 1438:1440 arg1 digests 1449:1455 the digests 1445:1455 the digests of bovine serum albumin 1445:1479 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 6 25 theme Ti 1057:1058 arg1 able 1086:1089 able 1086:1089 able 1086:1089 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 6 25 theme Ti 1057:1058 arg1 microspheres 1068:1079 the Ti(4+)-SPE microspheres 1053:1079 the Ti(4+)-SPE microspheres 1053:1079 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 8 26 theme mass 1486:1489 arg1 ratio 1491:1495 a mass ratio 1484:1495 a mass ratio of 1:100 1484:1504 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 4 27 theme MS 762:763 arg1 analysis 765:772 MS analysis 762:772 MS analysis of the N-glycome 762:789 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 7 28 gly glycoprotein 1291:1302 arg1 asialofetuin 1305:1316 asialofetuin 1305:1316 asialofetuin 1305:1316 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 28 gly glycoprotein 1291:1302 arg1 glycoprotein 1291:1302 a standard glycoprotein 1280:1302 a standard glycoprotein (asialofetuin, ASF) 1280:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 5 29 dep -SPE 892:895 arg1 extraction 910:919 solid phase extraction 898:919 Ti(4+)-SPE (solid phase extraction) 886:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 7 30 theme enrichment 1138:1147 arg1 method 1149:1154 The enrichment method 1134:1154 The enrichment method 1134:1154 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 31 gly sialylated 1236:1245 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 31 gly sialylated 1236:1245 arg1 A1 1254:1255 sialylated glycan A1 1236:1255 sialylated glycan A1 1236:1255 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 32 theme standard 1282:1289 arg1 asialofetuin 1305:1316 asialofetuin 1305:1316 asialofetuin 1305:1316 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 32 theme standard 1282:1289 arg1 glycoprotein 1291:1302 a standard glycoprotein 1280:1302 a standard glycoprotein (asialofetuin, ASF) 1280:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 8 33 from level 1610:1614 arg1 LOD 1598:1600 LOD 1598:1600 LOD at fmol level 1598:1614 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 33 from level 1610:1614 arg1 sensitivity 1585:1595 high sensitivity 1580:1595 high sensitivity (LOD at fmol level) 1580:1615 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 3 34 theme complex 466:472 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 0 35 theme N-glycans 32:40 arg1 purification 16:27 Highly specific purification 0:27 Highly specific purification of N-glycans 0:40 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 9 36 theme human 1705:1709 arg1 serum 1711:1715 human serum 1705:1715 human serum 1705:1715 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 6 37 theme reducing 1028:1035 arg1 such 1043:1046 such 1043:1046 such 1043:1046 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 6 37 theme reducing 1028:1035 arg1 ends 1037:1040 their reducing ends 1022:1040 their reducing ends 1022:1040 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 8 38 theme high 1508:1511 arg1 recovery 1523:1530 high enriching recovery 1508:1530 high enriching recovery (90%) 1508:1536 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 38 theme high 1508:1511 arg1 % 1535:1535 90% 1533:1535 90% 1533:1535 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 3 39 theme MS 663:664 arg1 analysis 666:673 the MS analysis 659:673 the MS analysis 659:673 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 3 40 theme basic 503:507 arg1 sites 509:513 basic sites 503:513 basic sites for protonation 503:529 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 5 41 theme N-glycans 808:816 arg1 approach 829:836 a novel N-glycans enrichment approach 800:836 a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) 800:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 8 42 dep selectivity 1415:1425 arg1 analyzing 1428:1436 analyzing 1428:1436 analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100 1428:1504 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 6 43 from ends 1037:1040 arg1 phospho-group 1005:1017 a phospho-group 1003:1017 a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans 1003:1131 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 3 44 theme lack 495:498 arg1 abundance 443:451 The relatively low abundance 424:451 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides 424:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 7 45 from glycoprotein 1291:1302 arg1 glycans 1267:1273 glycans 1267:1273 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 46 theme glycan 1247:1252 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 46 theme glycan 1247:1252 arg1 A1 1254:1255 sialylated glycan A1 1236:1255 sialylated glycan A1 1236:1255 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 1 47 theme variety 240:246 arg1 regulation 219:228 regulation 219:228 regulation of a wide variety of cellular processes 219:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 9 48 theme masses 1750:1755 arg1 total 1729:1733 a total 1727:1733 a total of 31 N-glycan masses 1727:1755 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 8 49 theme good 1539:1542 arg1 reproducibility 1544:1558 good reproducibility 1539:1558 good reproducibility (CV<15%) 1539:1567 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 49 theme good 1539:1542 arg1 % 1566:1566 CV<15% 1561:1566 CV<15% 1561:1566 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 3 50 theme other 551:555 arg1 proteins/peptides 573:589 other highly abundant proteins/peptides 551:589 other highly abundant proteins/peptides 551:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 1 51 theme cellular 251:258 arg1 processes 260:268 cellular processes 251:268 cellular processes 251:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 3 52 theme suppression 536:546 arg1 abundance 443:451 The relatively low abundance 424:451 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides 424:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 5 53 theme Ti 886:887 arg1 -SPE 892:895 Ti(4+)-SPE (solid phase extraction) 886:920 Ti(4+)-SPE (solid phase extraction) 886:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 0 54 theme phosphate-based 48:62 arg1 tag 94:96 an affinity tag 82:96 an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis 82:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 0 54 theme phosphate-based 48:62 arg1 derivatization 64:77 phosphate-based derivatization 48:77 phosphate-based derivatization 48:77 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 2 55 theme specific 316:323 arg1 identification 325:338 Highly specific identification 309:338 Highly specific identification of N-glycome 309:351 Highly specific identification of N-glycome remains a challenge while its biological significance is acknowledged. 27506354 8 56 theme fmol 1605:1608 arg1 level 1610:1614 fmol level 1605:1614 fmol level 1605:1614 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 1 57 theme N-linked 173:180 arg1 glycosylation 190:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation 173:202 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 3 58 theme abundant 564:571 arg1 proteins/peptides 573:589 other highly abundant proteins/peptides 551:589 other highly abundant proteins/peptides 551:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 5 59 theme phosphate 847:855 arg1 derivatization 857:870 phosphate derivatization 847:870 phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) 847:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 0 60 from tag 94:96 arg1 combination 101:111 combination 101:111 combination with Ti(4+)-SPE enrichment for mass spectrometric analysis 101:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 2 61 theme N-glycome 343:351 arg1 identification 325:338 Highly specific identification 309:338 Highly specific identification of N-glycome 309:351 Highly specific identification of N-glycome remains a challenge while its biological significance is acknowledged. 27506354 0 62 theme -SPE 124:127 arg1 enrichment 129:138 Ti(4+)-SPE enrichment 118:138 Ti(4+)-SPE enrichment for mass spectrometric analysis 118:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 3 63 theme poor 616:619 arg1 sensitivity 631:641 the particularly poor detection sensitivity 599:641 the particularly poor detection sensitivity of N-glycans 599:654 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 0 64 theme spectrometric 149:161 arg1 analysis 163:170 mass spectrometric analysis 144:170 mass spectrometric analysis 144:170 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 3 65 theme N-glycans 646:654 arg1 sensitivity 631:641 the particularly poor detection sensitivity 599:641 the particularly poor detection sensitivity of N-glycans 599:654 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 8 66 theme bovine 1460:1465 arg1 albumin 1473:1479 bovine serum albumin 1460:1479 bovine serum albumin 1460:1479 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 4 67 theme specific 698:705 arg1 procedure 720:728 the highly specific purification procedure 687:728 the highly specific purification procedure 687:728 Therefore, the highly specific purification procedure becomes a crucial step prior to MS analysis of the N-glycome. 27506354 3 68 from suppression 536:546 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 7 69 theme model 1190:1194 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 69 theme model 1190:1194 arg1 DP7 1228:1230 maltoheptaose DP7 1214:1230 maltoheptaose DP7 1214:1230 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 69 theme model 1190:1194 arg1 A1 1254:1255 sialylated glycan A1 1236:1255 sialylated glycan A1 1236:1255 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 5 70 theme solid 898:902 arg1 extraction 910:919 solid phase extraction 898:919 Ti(4+)-SPE (solid phase extraction) 886:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 8 71 theme albumin 1473:1479 arg1 digests 1449:1455 the digests 1445:1455 the digests of bovine serum albumin 1445:1479 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 5 72 theme enrichment 818:827 arg1 approach 829:836 a novel N-glycans enrichment approach 800:836 a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) 800:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 8 73 theme 1:100 1500:1504 arg1 ratio 1491:1495 a mass ratio 1484:1495 a mass ratio of 1:100 1484:1504 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 2 74 theme biological 383:392 arg1 significance 394:405 its biological significance 379:405 its biological significance 379:405 Highly specific identification of N-glycome remains a challenge while its biological significance is acknowledged. 27506354 9 75 theme N-glycome 1692:1700 arg1 profiling 1679:1687 the profiling 1675:1687 the profiling of N-glycome in human serum 1675:1715 At last, the proposed method was successfully applied in the profiling of N-glycome in human serum, in which a total of 31 N-glycan masses were identified. 27506354 7 76 dep oligosaccharides 1196:1211 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 76 dep oligosaccharides 1196:1211 arg1 DP7 1228:1230 maltoheptaose DP7 1214:1230 maltoheptaose DP7 1214:1230 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 76 dep oligosaccharides 1196:1211 arg1 A1 1254:1255 sialylated glycan A1 1236:1255 sialylated glycan A1 1236:1255 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 8 77 from ratio 1491:1495 arg1 DP7 1438:1440 DP7 1438:1440 DP7 in the digests of bovine serum albumin at a mass ratio of 1:100 1438:1504 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 1 78 theme numerous 290:297 arg1 diseases 299:306 numerous diseases 290:306 numerous diseases 290:306 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 3 79 theme low 439:441 arg1 abundance 443:451 The relatively low abundance 424:451 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides 424:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 8 80 theme enriching 1513:1521 arg1 recovery 1523:1530 high enriching recovery 1508:1530 high enriching recovery (90%) 1508:1536 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 80 theme enriching 1513:1521 arg1 % 1535:1535 90% 1533:1535 90% 1533:1535 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 81 theme derivatization 1364:1377 arg1 % 1401:1401 almost 100% 1391:1401 almost 100% 1391:1401 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 81 theme derivatization 1364:1377 arg1 efficiency 1379:1388 high derivatization efficiency 1359:1388 high derivatization efficiency (almost 100%) 1359:1402 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 3 82 theme biological 474:483 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 5 83 theme novel 802:806 arg1 approach 829:836 a novel N-glycans enrichment approach 800:836 a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) 800:920 Herein, a novel N-glycans enrichment approach based on phosphate derivatization combined with Ti(4+)-SPE (solid phase extraction) was developed. 27506354 7 84 dep asialofetuin 1305:1316 arg1 ASF 1319:1321 ASF 1319:1321 ASF 1319:1321 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 3 85 from mixtures 485:492 arg1 abundance 443:451 The relatively low abundance 424:451 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides 424:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 3 86 theme sites 509:513 arg1 glycan 456:461 glycan 456:461 glycan in complex biological mixtures 456:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 3 86 theme sites 509:513 arg1 lack 495:498 lack 495:498 lack of basic sites for protonation 495:529 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 3 86 theme sites 509:513 arg1 suppression 536:546 suppression 536:546 suppression by other highly abundant proteins/peptides 536:589 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 3 87 from abundance 443:451 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 6 88 theme -SPE 1063:1066 arg1 able 1086:1089 able 1086:1089 able 1086:1089 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 6 88 theme -SPE 1063:1066 arg1 microspheres 1068:1079 the Ti(4+)-SPE microspheres 1053:1079 the Ti(4+)-SPE microspheres 1053:1079 Briefly, in this strategy, N-glycans were chemically labeled with a phospho-group at their reducing ends, such that the Ti(4+)-SPE microspheres were able to capture the phospho-containing glycans. 27506354 7 89 theme sialylated 1236:1245 arg1 oligosaccharides 1196:1211 model oligosaccharides 1190:1211 model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF) 1190:1322 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 7 89 theme sialylated 1236:1245 arg1 A1 1254:1255 sialylated glycan A1 1236:1255 sialylated glycan A1 1236:1255 The enrichment method was developed and optimized using model oligosaccharides (maltoheptaose DP7 and sialylated glycan A1) and also glycans from a standard glycoprotein (asialofetuin, ASF). 27506354 3 90 from glycan 456:461 arg1 mixtures 485:492 complex biological mixtures 466:492 complex biological mixtures 466:492 The relatively low abundance of glycan in complex biological mixtures, lack of basic sites for protonation, and suppression by other highly abundant proteins/peptides lead to the particularly poor detection sensitivity of N-glycans in the MS analysis. 27506354 0 91 theme specific 7:14 arg1 purification 16:27 Highly specific purification 0:27 Highly specific purification of N-glycans 0:40 Highly specific purification of N-glycans using phosphate-based derivatization as an affinity tag in combination with Ti(4+)-SPE enrichment for mass spectrometric analysis. 27506354 8 92 theme high 1580:1583 arg1 LOD 1598:1600 LOD 1598:1600 LOD at fmol level 1598:1614 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 8 92 theme high 1580:1583 arg1 sensitivity 1585:1595 high sensitivity 1580:1595 high sensitivity (LOD at fmol level) 1580:1615 This method experimentally showed high derivatization efficiency (almost 100%), excellent selectivity (analyzing DP7 in the digests of bovine serum albumin at a mass ratio of 1:100), high enriching recovery (90%), good reproducibility (CV<15%) as well as high sensitivity (LOD at fmol level). 27506354 1 93 theme wide 235:238 arg1 variety 240:246 a wide variety 233:246 a wide variety of cellular processes 233:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 27506354 1 93 theme wide 235:238 arg1 processes 260:268 cellular processes 251:268 cellular processes 251:268 N-linked protein glycosylation is involved in regulation of a wide variety of cellular processes and associated with numerous diseases. 28585084 3 0 theme published 711:719 arg1 literature 721:730 published literature 711:730 published literature 711:730 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 5 1 theme glycomics 1066:1074 arg1 subset 1051:1056 subset 1051:1056 subset of both glycomics and proteomics 1051:1089 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 4 2 theme glycans 885:891 arg1 function 869:876 function 869:876 function 869:876 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 4 2 theme glycans 885:891 arg1 structure 855:863 structure 855:863 structure 855:863 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 2 3 from fields 313:318 arg1 researchers 292:302 researchers 292:302 researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure 292:498 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 2 4 theme biology 333:339 arg1 fields 313:318 the fields 309:318 the fields of molecular biology and biochemistry 309:356 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 3 5 theme references 697:706 arg1 explanations 641:652 explanations 641:652 explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 641:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 2 6 dep tools 465:469 arg1 elucidate 474:482 elucidate 474:482 to elucidate their structure 471:498 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 4 7 theme structure 855:863 arg1 study 842:846 the comprehensive study 824:846 the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases 824:1023 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 1 8 gly glycoproteins 93:105 arg1 glycoproteins 93:105 glycoproteins 93:105 glycoproteins 93:105 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 5 9 theme proteins 1137:1144 arg1 characterization 1117:1132 characterization 1117:1132 characterization 1117:1132 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 5 9 theme proteins 1137:1144 arg1 identification 1098:1111 identification 1098:1111 identification 1098:1111 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 3 10 theme major 539:543 arg1 techniques 572:581 the major and most common analytical techniques 535:581 the major and most common analytical techniques 535:581 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 10 theme major 539:543 arg1 explanations 641:652 explanations 641:652 explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 641:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 1 11 theme structural 70:79 arg1 analysis 81:88 The structural analysis 66:88 The structural analysis of glycoproteins 66:105 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 1 11 theme structural 70:79 arg1 endeavor 124:131 a challenging endeavor 110:131 a challenging endeavor 110:131 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 2 12 theme molecular 323:331 arg1 biology 333:339 molecular biology 323:339 molecular biology 323:339 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 4 13 theme function 869:876 arg1 study 842:846 the comprehensive study 824:846 the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases 824:1023 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 4 14 theme genes 968:972 arg1 identification 942:955 identification 942:955 identification of all the genes that encode glycoproteins and glycosyltransferases 942:1023 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 2 15 gly glycoproteins 384:396 arg1 glycoproteins 384:396 glycoproteins 384:396 glycoproteins 384:396 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 6 16 theme ease 1232:1235 arg1 entry 1237:1241 ease entry 1232:1241 ease entry into the glycomics and glycoproteomics field 1232:1286 This tutorial is designed to ease entry into the glycomics and glycoproteomics field for those without prior carbohydrate analysis experience. 28585084 3 17 theme brief 513:517 arg1 descriptions 519:530 brief descriptions 513:530 brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 513:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 1 18 theme glycoproteins 93:105 arg1 analysis 81:88 The structural analysis 66:88 The structural analysis of glycoproteins 66:105 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 1 18 theme glycoproteins 93:105 arg1 endeavor 124:131 a challenging endeavor 110:131 a challenging endeavor 110:131 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 3 19 theme experimental 747:758 arg1 details 760:766 the experimental details 743:766 the experimental details necessary to carry out the analyses 743:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 6 20 theme carbohydrate 1312:1323 arg1 analysis 1325:1332 prior carbohydrate analysis 1306:1332 prior carbohydrate analysis 1306:1332 This tutorial is designed to ease entry into the glycomics and glycoproteomics field for those without prior carbohydrate analysis experience. 28585084 3 21 theme analytical 561:570 arg1 techniques 572:581 the major and most common analytical techniques 535:581 the major and most common analytical techniques 535:581 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 21 theme analytical 561:570 arg1 explanations 641:652 explanations 641:652 explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 641:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 22 theme common 554:559 arg1 techniques 572:581 the major and most common analytical techniques 535:581 the major and most common analytical techniques 535:581 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 22 theme common 554:559 arg1 explanations 641:652 explanations 641:652 explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 641:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 1 23 theme limited 190:196 arg1 number 198:203 only a very limited number 178:203 only a very limited number of labs 178:211 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 2 24 theme biological 421:430 arg1 research 432:439 their biological research 415:439 their biological research 415:439 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 5 25 theme posttranslational 1171:1187 arg1 modification 1189:1200 posttranslational modification 1171:1200 posttranslational modification 1171:1200 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 1 26 theme challenging 112:122 arg1 analysis 81:88 The structural analysis 66:88 The structural analysis of glycoproteins 66:105 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 1 26 theme challenging 112:122 arg1 endeavor 124:131 a challenging endeavor 110:131 a challenging endeavor 110:131 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 0 27 theme glycoproteins-a 40:54 arg1 tutorial 56:63 glycoproteins-a tutorial 40:63 glycoproteins-a tutorial 40:63 Glycomic and glycoproteomic analysis of glycoproteins-a tutorial. 28585084 3 28 theme necessary 768:776 arg1 details 760:766 the experimental details 743:766 the experimental details necessary to carry out the analyses 743:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 4 29 gly glycoproteins 986:998 arg1 genes 968:972 all the genes 960:972 all the genes that encode glycoproteins and glycosyltransferases 960:1023 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 4 29 gly glycoproteins 986:998 arg1 glycoproteins 986:998 glycoproteins 986:998 glycoproteins 986:998 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 6 30 theme prior 1306:1310 arg1 analysis 1325:1332 prior carbohydrate analysis 1306:1332 prior carbohydrate analysis 1306:1332 This tutorial is designed to ease entry into the glycomics and glycoproteomics field for those without prior carbohydrate analysis experience. 28585084 6 31 theme glycomics 1252:1260 arg1 field 1282:1286 the glycomics and glycoproteomics field 1248:1286 field 1282:1286 This tutorial is designed to ease entry into the glycomics and glycoproteomics field for those without prior carbohydrate analysis experience. 28585084 1 32 theme labs 208:211 arg1 number 198:203 only a very limited number 178:203 only a very limited number of labs 178:211 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 3 33 theme techniques 572:581 arg1 descriptions 519:530 brief descriptions 513:530 brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 513:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 34 contain containing 732:741 arg2 details 760:766 the experimental details 743:766 the experimental details necessary to carry out the analyses 743:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 3 34 contain containing 732:741 arg1 rationales 661:670 the rationales 657:670 the rationales for individual steps 657:691 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 0 35 theme tutorial 56:63 arg1 Glycomic 0:7 Glycomic 0:7 Glycomic 0:7 Glycomic and glycoproteomic analysis of glycoproteins-a tutorial. 28585084 0 35 theme tutorial 56:63 arg1 analysis 28:35 glycoproteomic analysis 13:35 glycoproteomic analysis of glycoproteins-a tutorial 13:63 Glycomic and glycoproteomic analysis of glycoproteins-a tutorial. 28585084 4 36 theme given 908:912 arg1 cell 914:917 a given cell 906:917 a given cell 906:917 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 3 37 theme individual 676:685 arg1 steps 687:691 individual steps 676:691 individual steps 676:691 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 1 38 contain have 213:216 arg1 number 198:203 only a very limited number 178:203 only a very limited number of labs 178:211 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 1 38 contain have 213:216 arg2 expertise 222:230 the expertise 218:230 the expertise required to accomplish this task 218:263 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 3 39 theme glycoproteomics 605:619 arg1 analysis 621:628 glycoproteomics analysis 605:628 glycoproteomics analysis 605:628 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 5 40 theme proteomics 1080:1089 arg1 subset 1051:1056 subset 1051:1056 subset of both glycomics and proteomics 1051:1089 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 3 41 theme rationales 661:670 arg1 explanations 641:652 explanations 641:652 explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses 641:802 It provides brief descriptions of the major and most common analytical techniques used in glycomics and glycoproteomics analysis, including explanations of the rationales for individual steps and references to published literature containing the experimental details necessary to carry out the analyses. 28585084 6 42 theme glycoproteomics 1266:1280 arg1 field 1282:1286 the glycomics and glycoproteomics field 1248:1286 field 1282:1286 This tutorial is designed to ease entry into the glycomics and glycoproteomics field for those without prior carbohydrate analysis experience. 28585084 2 43 from important 402:410 arg1 research 432:439 their biological research 415:439 their biological research 415:439 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 1 44 theme increasing 155:164 arg1 demand 166:171 steadily increasing demand 146:171 steadily increasing demand 146:171 The structural analysis of glycoproteins is a challenging endeavor and is under steadily increasing demand, but only a very limited number of labs have the expertise required to accomplish this task. 28585084 4 45 theme comprehensive 828:840 arg1 study 842:846 the comprehensive study 824:846 the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases 824:1023 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 4 46 dep structure 855:863 arg1 the 851:853 the 851:853 the 851:853 Glycomics includes the comprehensive study of the structure and function of the glycans expressed in a given cell or organism along with identification of all the genes that encode glycoproteins and glycosyltransferases. 28585084 5 47 dep identification 1098:1111 arg1 the 1094:1096 the 1094:1096 the 1094:1096 Glycoproteomics which is subset of both glycomics and proteomics is the identification and characterization of proteins bearing carbohydrates as posttranslational modification. 28585084 2 48 from research 432:439 arg1 important 402:410 important 402:410 important 402:410 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 28585084 0 49 theme glycoproteomic 13:26 arg1 analysis 28:35 glycoproteomic analysis 13:35 glycoproteomic analysis of glycoproteins-a tutorial 13:63 Glycomic and glycoproteomic analysis of glycoproteins-a tutorial. 28585084 2 50 theme biochemistry 345:356 arg1 fields 313:318 the fields 309:318 the fields of molecular biology and biochemistry 309:356 This tutorial is aimed at researchers from the fields of molecular biology and biochemistry that have discovered that glycoproteins are important in their biological research and are looking for the tools to elucidate their structure. 26056814 8 0 theme binding 1690:1696 arg1 affinity 1698:1705 the overall glycan binding affinity 1671:1705 the overall glycan binding affinity of the HA 1671:1715 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 3 1 theme A/California/07/09 679:696 arg1 selectivity 586:596 The glycan-binding selectivity 567:596 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations 567:726 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 1 2 theme host 271:274 arg1 range 276:280 the host range 267:280 the host range of the virus and egg-adaption during vaccine production 267:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 6 3 theme binding 1236:1242 arg1 affinity 1244:1251 an enhanced binding affinity 1224:1251 an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans 1224:1297 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 10 4 theme vaccine 1974:1980 arg1 strategies 1989:1998 future vaccine design strategies 1967:1998 future vaccine design strategies 1967:1998 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 3 5 theme ELISA 759:763 arg1 assay 765:769 a solid-phase ELISA assay 745:769 a solid-phase ELISA assay 745:769 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 1 6 theme binding 189:195 arg1 determinant 251:261 an important determinant 238:261 an important determinant for the host range of the virus and egg-adaption during vaccine production 238:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 1 6 theme binding 189:195 arg1 selectivity 197:207 The haemagglutinin (HA) glycan binding selectivity 158:207 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses 158:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 4 7 link α2,3-linked 891:901 arg1 sialyl-glycans 903:916 the α2,3-linked sialyl-glycans 887:916 the α2,3-linked sialyl-glycans in the array 887:929 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 0 8 theme Swine 133:137 arg1 Virus 151:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 8 9 gly glycosylation 1544:1556 arg2 site 1558:1561 a glycosylation site 1542:1561 a glycosylation site proximal to the receptor binding site 1542:1599 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 8 10 theme K123N 1510:1514 arg1 mutation 1516:1523 The K123N mutation 1506:1523 The K123N mutation 1506:1523 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 10 11 theme H1N1 2074:2077 arg1 viruses 2079:2085 recombinant H1N1 viruses 2062:2085 recombinant H1N1 viruses 2062:2085 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 0 12 theme A 149:149 arg1 Virus 151:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 1 13 theme influenza 217:225 arg1 viruses 227:233 H1N1 influenza viruses 212:233 H1N1 influenza viruses 212:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 2 14 theme A 557:557 arg1 virus 559:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 6 15 link α2,3-linked 1272:1282 arg1 sialyl-glycans 1284:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 5 16 theme binding 1072:1078 arg1 pattern 1045:1051 a comparable pattern 1032:1051 a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans 1032:1108 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 5 17 link α2,3-linked 1083:1093 arg1 sialyl-glycans 1095:1108 α2,3-linked sialyl-glycans 1083:1108 α2,3-linked sialyl-glycans 1083:1108 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 2 18 theme swine 541:545 arg1 virus 559:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 6 19 theme α2,3-linked 1272:1282 arg1 sialyl-glycans 1284:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 5 20 theme α2,3-linked 1083:1093 arg1 sialyl-glycans 1095:1108 α2,3-linked sialyl-glycans 1083:1108 α2,3-linked sialyl-glycans 1083:1108 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 5 21 theme virus 957:961 arg1 strains 963:969 vaccine virus strains 949:969 vaccine virus strains 949:969 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 9 22 theme residues 1804:1811 arg1 scoring 1814:1820 'per residues' scoring 1799:1820 'per residues' scoring 1799:1820 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 6 23 theme α2,6- 1262:1266 arg1 sialyl-glycans 1284:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 both α2,6- and α2,3-linked sialyl-glycans 1257:1297 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 0 24 theme Egg-Adapted 79:89 arg1 Strains 99:105 Egg-Adapted Vaccine Strains 79:105 Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 79:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 1 25 dep virus 289:293 arg1 the 285:287 the 285:287 the 285:287 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 9 26 theme per 1800:1802 arg1 residues 1804:1811 per residues 1800:1811 'per residues' scoring 1799:1820 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 5 27 dep strains 963:969 arg1 the 945:947 the 945:947 the 945:947 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 0 28 theme Strains 99:105 arg1 Specificity 64:74 the Haemagglutinin Glycan-Binding Specificity 30:74 the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 30:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 4 29 from sialyl-glycans 903:916 arg1 array 925:929 the array 921:929 the array 921:929 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 2 30 theme strains 507:513 arg1 HA 493:494 the HA 489:494 the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus 489:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 8 31 theme receptor 1579:1586 arg1 site 1596:1599 the receptor binding site 1575:1599 the receptor binding site 1575:1599 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 4 32 theme Wild-type 772:780 arg1 HA 813:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 2 33 dep mutations 467:475 arg1 seen 481:484 seen 481:484 seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus 481:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 5 34 link α2,6-linked 1142:1152 arg1 sialyl-glycans 1154:1167 α2,6-linked sialyl-glycans 1142:1167 α2,6-linked sialyl-glycans 1142:1167 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 4 35 theme recombinant 801:811 arg1 HA 813:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 5 36 theme α2,6-linked 1142:1152 arg1 sialyl-glycans 1154:1167 α2,6-linked sialyl-glycans 1142:1167 α2,6-linked sialyl-glycans 1142:1167 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 5 37 theme HA 1010:1011 arg1 mutation 1013:1020 the Q226R HA mutation 1000:1020 the Q226R HA mutation 1000:1020 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 2 38 theme D225G 451:455 arg1 mutations 467:475 the K123N, D225G and Q226R mutations 440:475 the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus) 440:564 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 2 39 theme structure-recognition 386:406 arg1 models 408:413 structure-recognition models 386:413 structure-recognition models 386:413 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 0 40 theme Molecular 0:8 arg1 Characterisation 10:25 Molecular Characterisation 0:25 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 0:156 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 3 41 dep A/California/07/09 679:696 arg1 HAs 698:700 HAs 698:700 HAs 698:700 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 4 42 link α2,6-linked 838:848 arg1 sialyl-glycans 850:863 α2,6-linked sialyl-glycans 838:863 α2,6-linked sialyl-glycans 838:863 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 7 43 theme α2,3-glycan 1383:1393 arg1 affinity 1395:1402 its α2,3-glycan affinity 1379:1402 its α2,3-glycan affinity 1379:1402 Notably its α2,6-glycan affinity was generally higher compared to its α2,3-glycan affinity, which may explain why the double mutant was not naturally selected during egg-adaption of the virus. 26056814 9 44 theme experimental 1888:1899 arg1 data 1916:1919 the experimental glycan binding data 1884:1919 the experimental glycan binding data 1884:1919 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 8 45 theme α2,3/α2,6 1621:1629 arg1 selectivity 1638:1648 the α2,3/α2,6 glycan selectivity 1617:1648 the α2,3/α2,6 glycan selectivity 1617:1648 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 3 46 theme strains 645:651 arg1 selectivity 586:596 The glycan-binding selectivity 567:596 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations 567:726 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 0 47 theme Haemagglutinin 34:47 arg1 Specificity 64:74 the Haemagglutinin Glycan-Binding Specificity 30:74 the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 30:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 9 48 theme glycan 1901:1906 arg1 data 1916:1919 the experimental glycan binding data 1884:1919 the experimental glycan binding data 1884:1919 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 2 49 theme Q226R 461:465 arg1 mutations 467:475 the K123N, D225G and Q226R mutations 440:475 the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus) 440:564 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 3 50 theme vaccine 626:632 arg1 strains 645:651 three A/California/07/09 vaccine production strains 601:651 three A/California/07/09 vaccine production strains 601:651 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 0 51 theme Specificity 64:74 arg1 Characterisation 10:25 Molecular Characterisation 0:25 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 0:156 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 5 52 theme specific 1063:1070 arg1 binding 1072:1078 highly specific binding 1056:1078 highly specific binding to α2,3-linked sialyl-glycans 1056:1108 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 3 53 theme recombinant 667:677 arg1 A/California/07/09 679:696 purified recombinant A/California/07/09 HAs 658:700 purified recombinant A/California/07/09 HAs harboring these mutations 658:726 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 1 54 theme viruses 227:233 arg1 determinant 251:261 an important determinant 238:261 an important determinant for the host range of the virus and egg-adaption during vaccine production 238:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 1 54 theme viruses 227:233 arg1 selectivity 197:207 The haemagglutinin (HA) glycan binding selectivity 158:207 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses 158:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 7 55 theme virus 1499:1503 arg1 egg-adaption 1479:1490 egg-adaption 1479:1490 egg-adaption of the virus 1479:1503 Notably its α2,6-glycan affinity was generally higher compared to its α2,3-glycan affinity, which may explain why the double mutant was not naturally selected during egg-adaption of the virus. 26056814 2 56 with data 376:379 arg1 models 408:413 structure-recognition models 386:413 structure-recognition models 386:413 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 10 57 theme glycan 1940:1945 arg1 data 1955:1958 the glycan binding data 1936:1958 the glycan binding data 1936:1958 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 9 58 theme binding 1908:1914 arg1 data 1916:1919 the experimental glycan binding data 1884:1919 the experimental glycan binding data 1884:1919 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 5 59 theme comparable 1034:1043 arg1 pattern 1045:1051 a comparable pattern 1032:1051 a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans 1032:1108 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 7 60 theme α2,6-glycan 1325:1335 arg1 affinity 1337:1344 its α2,6-glycan affinity 1321:1344 its α2,6-glycan affinity 1321:1344 Notably its α2,6-glycan affinity was generally higher compared to its α2,3-glycan affinity, which may explain why the double mutant was not naturally selected during egg-adaption of the virus. 26056814 10 61 theme design 1982:1987 arg1 strategies 1989:1998 future vaccine design strategies 1967:1998 future vaccine design strategies 1967:1998 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 2 62 theme glycan 361:366 arg1 data 376:379 glycan binding data 361:379 glycan binding data with structure-recognition models 361:413 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 4 63 theme α2,3-linked 891:901 arg1 sialyl-glycans 903:916 the α2,3-linked sialyl-glycans 887:916 the α2,3-linked sialyl-glycans in the array 887:929 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 8 64 theme glycan 1683:1688 arg1 affinity 1698:1705 the overall glycan binding affinity 1671:1705 the overall glycan binding affinity of the HA 1671:1715 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 10 65 theme future 1967:1972 arg1 strategies 1989:1998 future vaccine design strategies 1967:1998 future vaccine design strategies 1967:1998 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 2 66 theme virus 559:563 arg1 strains 507:513 vaccine strains 499:513 vaccine strains of the pandemic 2009 H1N1 swine influenza A virus 499:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 8 67 theme receptor 1762:1769 arg1 binding 1771:1777 receptor binding 1762:1777 receptor binding 1762:1777 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 6 68 theme enhanced 1227:1234 arg1 affinity 1244:1251 an enhanced binding affinity 1224:1251 an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans 1224:1297 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 10 69 theme acid 2038:2041 arg1 substitutions 2043:2055 amino acid substitutions 2032:2055 amino acid substitutions 2032:2055 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 1 70 theme glycan 182:187 arg1 determinant 251:261 an important determinant 238:261 an important determinant for the host range of the virus and egg-adaption during vaccine production 238:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 1 70 theme glycan 182:187 arg1 selectivity 197:207 The haemagglutinin (HA) glycan binding selectivity 158:207 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses 158:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 10 71 theme amino 2032:2036 arg1 substitutions 2043:2055 amino acid substitutions 2032:2055 amino acid substitutions 2032:2055 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 0 72 theme Influenza 139:147 arg1 Virus 151:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 2 73 theme pandemic 522:529 arg1 virus 559:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 6 74 theme recombinant 1199:1209 arg1 HA 1211:1212 The D225G A/California/07/09 recombinant HA 1170:1212 The D225G A/California/07/09 recombinant HA 1170:1212 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 3 75 theme solid-phase 747:757 arg1 assay 765:769 a solid-phase ELISA assay 745:769 a solid-phase ELISA assay 745:769 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 0 76 theme Virus 151:155 arg1 Strains 99:105 Egg-Adapted Vaccine Strains 79:105 Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 79:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 1 77 theme H1N1 212:215 arg1 viruses 227:233 H1N1 influenza viruses 212:233 H1N1 influenza viruses 212:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 9 78 theme Docking 1780:1786 arg1 models 1788:1793 Docking models 1780:1793 Docking models 1780:1793 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 10 79 theme recombinant 2062:2072 arg1 viruses 2079:2085 recombinant H1N1 viruses 2062:2085 recombinant H1N1 viruses 2062:2085 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 5 80 theme negligible 1118:1127 arg1 affinity 1129:1136 a negligible affinity 1116:1136 a negligible affinity for α2,6-linked sialyl-glycans 1116:1167 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 2 81 theme influenza 547:555 arg1 virus 559:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 8 82 theme glycosylation 1544:1556 arg1 site 1558:1561 a glycosylation site 1542:1561 a glycosylation site proximal to the receptor binding site 1542:1599 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 5 83 theme vaccine 949:955 arg1 strains 963:969 vaccine virus strains 949:969 vaccine virus strains 949:969 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 2 84 theme H1N1 536:539 arg1 virus 559:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 the pandemic 2009 H1N1 swine influenza A virus 518:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 8 85 theme proximal 1563:1570 arg1 site 1558:1561 a glycosylation site 1542:1561 a glycosylation site proximal to the receptor binding site 1542:1599 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 7 86 theme double 1431:1436 arg1 mutant 1438:1443 the double mutant 1427:1443 the double mutant 1427:1443 Notably its α2,6-glycan affinity was generally higher compared to its α2,3-glycan affinity, which may explain why the double mutant was not naturally selected during egg-adaption of the virus. 26056814 0 87 theme Vaccine 91:97 arg1 Strains 99:105 Egg-Adapted Vaccine Strains 79:105 Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 79:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 3 88 theme glycan-binding 571:584 arg1 selectivity 586:596 The glycan-binding selectivity 567:596 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations 567:726 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 1 89 theme haemagglutinin 162:175 arg1 determinant 251:261 an important determinant 238:261 an important determinant for the host range of the virus and egg-adaption during vaccine production 238:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 1 89 theme haemagglutinin 162:175 arg1 selectivity 197:207 The haemagglutinin (HA) glycan binding selectivity 158:207 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses 158:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 8 90 theme binding 1588:1594 arg1 site 1596:1599 the receptor binding site 1575:1599 the receptor binding site 1575:1599 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 0 91 theme Pandemic 114:121 arg1 Virus 151:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 2 92 theme vaccine 499:505 arg1 strains 507:513 vaccine strains 499:513 vaccine strains of the pandemic 2009 H1N1 swine influenza A virus 499:563 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 0 93 theme H1N1 128:131 arg1 Virus 151:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 the Pandemic 2009 H1N1 Swine Influenza A Virus 110:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 5 94 theme Q226R 1004:1008 arg1 mutation 1013:1020 the Q226R HA mutation 1000:1020 the Q226R HA mutation 1000:1020 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 4 95 theme A/California/07/09 782:799 arg1 HA 813:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA 772:814 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 9 96 theme rational 1871:1878 arg1 structure-recognition 1849:1869 a structure-recognition 1847:1869 a structure-recognition rational for the experimental glycan binding data 1847:1919 Docking models and 'per residues' scoring were employed to provide a structure-recognition rational for the experimental glycan binding data. 26056814 5 97 theme recombinant 975:985 arg1 HA 987:988 recombinant HA 975:988 recombinant HA 975:988 In contrast, the vaccine virus strains and recombinant HA harboring the Q226R HA mutation displayed a comparable pattern of highly specific binding to α2,3-linked sialyl-glycans, with a negligible affinity for α2,6-linked sialyl-glycans. 26056814 8 98 theme glycan 1631:1636 arg1 selectivity 1638:1648 the α2,3/α2,6 glycan selectivity 1617:1648 the α2,3/α2,6 glycan selectivity 1617:1648 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 3 99 theme production 634:643 arg1 strains 645:651 three A/California/07/09 vaccine production strains 601:651 three A/California/07/09 vaccine production strains 601:651 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 0 100 theme Glycan-Binding 49:62 arg1 Specificity 64:74 the Haemagglutinin Glycan-Binding Specificity 30:74 the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus 30:155 Molecular Characterisation of the Haemagglutinin Glycan-Binding Specificity of Egg-Adapted Vaccine Strains of the Pandemic 2009 H1N1 Swine Influenza A Virus. 26056814 1 101 theme virus 289:293 arg1 range 276:280 the host range 267:280 the host range of the virus and egg-adaption during vaccine production 267:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 2 102 theme mutations 467:475 arg1 impact 430:435 the impact 426:435 the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus) 426:564 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 3 103 theme A/California/07/09 607:624 arg1 strains 645:651 three A/California/07/09 vaccine production strains 601:651 three A/California/07/09 vaccine production strains 601:651 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 4 104 theme α2,6-linked 838:848 arg1 sialyl-glycans 850:863 α2,6-linked sialyl-glycans 838:863 α2,6-linked sialyl-glycans 838:863 Wild-type A/California/07/09 recombinant HA bound specifically to α2,6-linked sialyl-glycans, with no affinity for the α2,3-linked sialyl-glycans in the array. 26056814 1 105 theme egg-adaption 299:310 arg1 range 276:280 the host range 267:280 the host range of the virus and egg-adaption during vaccine production 267:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 6 106 theme A/California/07/09 1180:1197 arg1 HA 1211:1212 The D225G A/California/07/09 recombinant HA 1170:1212 The D225G A/California/07/09 recombinant HA 1170:1212 The D225G A/California/07/09 recombinant HA displayed an enhanced binding affinity for both α2,6- and α2,3-linked sialyl-glycans in the array. 26056814 3 107 theme purified 658:665 arg1 A/California/07/09 679:696 purified recombinant A/California/07/09 HAs 658:700 purified recombinant A/California/07/09 HAs harboring these mutations 658:726 The glycan-binding selectivity of three A/California/07/09 vaccine production strains, and purified recombinant A/California/07/09 HAs harboring these mutations was examined via a solid-phase ELISA assay. 26056814 8 108 theme HA 1714:1715 arg1 affinity 1698:1705 the overall glycan binding affinity 1671:1705 the overall glycan binding affinity of the HA 1671:1715 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 1 109 theme vaccine 319:325 arg1 production 327:336 vaccine production 319:336 vaccine production 319:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 10 110 dep D225G 2017:2021 arg1 the 2013:2015 the 2013:2015 the 2013:2015 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 10 110 dep D225G 2017:2021 arg1 substitutions 2043:2055 amino acid substitutions 2032:2055 amino acid substitutions 2032:2055 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 10 111 theme binding 1947:1953 arg1 data 1955:1958 the glycan binding data 1936:1958 the glycan binding data 1936:1958 Collectively, the glycan binding data inform future vaccine design strategies to introduce the D225G or Q226R amino acid substitutions into recombinant H1N1 viruses. 26056814 8 112 theme overall 1675:1681 arg1 affinity 1698:1705 the overall glycan binding affinity 1671:1705 the overall glycan binding affinity of the HA 1671:1715 The K123N mutation which introduces a glycosylation site proximal to the receptor binding site, did not impact the α2,3/α2,6 glycan selectivity, however, it lowered the overall glycan binding affinity of the HA; suggesting glycosylation may interfere with receptor binding. 26056814 2 113 theme binding 368:374 arg1 data 376:379 glycan binding data 361:379 glycan binding data with structure-recognition models 361:413 This study integrates glycan binding data with structure-recognition models to examine the impact of the K123N, D225G and Q226R mutations (as seen in the HA of vaccine strains of the pandemic 2009 H1N1 swine influenza A virus). 26056814 1 114 theme important 241:249 arg1 selectivity 197:207 The haemagglutinin (HA) glycan binding selectivity 158:207 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses 158:233 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 26056814 1 114 theme important 241:249 arg1 determinant 251:261 an important determinant 238:261 an important determinant for the host range of the virus and egg-adaption during vaccine production 238:336 The haemagglutinin (HA) glycan binding selectivity of H1N1 influenza viruses is an important determinant for the host range of the virus and egg-adaption during vaccine production. 28443077 0 0 theme Trimer 69:74 arg1 Simulation 19:28 Molecular Dynamics Simulation 0:28 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer 0:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 1 theme molecular 669:677 arg1 simulations 688:698 molecular dynamics simulations 669:698 molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 669:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 2 theme Influenza 164:172 arg1 A 174:174 the Recent Epidemic Clade 3C.2a. Influenza A 131:174 the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 131:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 2 theme Influenza 164:172 arg1 H3N2 176:179 H3N2 176:179 H3N2 176:179 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 3 theme nonhuman 903:910 arg1 erythrocytes 912:923 nonhuman erythrocytes 903:923 nonhuman erythrocytes 903:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 4 from influenza 217:225 arg1 humans 230:235 humans 230:235 humans since 1968 230:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 5 theme HA 988:989 arg1 trimers 991:997 the glycosylated HA trimers 971:997 the glycosylated HA trimers 971:997 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 5 6 theme ligand 1335:1340 arg1 specificity 1350:1360 the ligand binding specificity 1331:1360 the ligand binding specificity 1331:1360 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 0 7 theme Hemagglutinin 55:67 arg1 Trimer 69:74 the Influenza A(H3N2) Hemagglutinin Trimer 33:74 the Influenza A(H3N2) Hemagglutinin Trimer 33:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 8 theme 3C.2a. 157:162 arg1 A 174:174 the Recent Epidemic Clade 3C.2a. Influenza A 131:174 the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 131:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 8 theme 3C.2a. 157:162 arg1 H3N2 176:179 H3N2 176:179 H3N2 176:179 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 9 theme major 193:197 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 9 theme major 193:197 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 5 10 theme 3C.2a 1210:1214 arg1 strain 1216:1221 the 3C.2a strain 1206:1221 the 3C.2a strain 1206:1221 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 2 11 theme structural 574:583 arg1 basis 585:589 the structural basis 570:589 the structural basis of these changes 570:606 However, the structural basis of these changes remains largely unclear. 28443077 5 12 theme HA 1238:1239 arg1 structure 1241:1249 an HA structure 1235:1249 an HA structure that is advantageous for evading pre-existing antibodies 1235:1306 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 5 12 theme HA 1238:1239 arg1 advantageous 1259:1270 advantageous 1259:1270 advantageous 1259:1270 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 3 13 from 3C.2a 839:843 arg1 unique 798:803 unique 798:803 unique 798:803 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 6 14 theme changes 1447:1453 arg1 understanding 1415:1427 our understanding 1411:1427 our understanding of the phenotypic changes 1411:1453 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 14 theme changes 1447:1453 arg1 evolution 1456:1464 evolution 1456:1464 evolution 1456:1464 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 14 theme changes 1447:1453 arg1 fate 1471:1474 fate 1471:1474 fate 1471:1474 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 0 15 dep Basis 99:103 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 15 dep Basis 99:103 arg1 evolved 257:263 evolved 257:263 has evolved by antigenic drift under the constantly changing human herd immunity 253:332 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 15 dep Basis 99:103 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 5 16 theme pre-existing 1284:1295 arg1 antibodies 1297:1306 pre-existing antibodies 1284:1306 pre-existing antibodies 1284:1306 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 0 17 theme A 174:174 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 17 theme A 174:174 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 6 18 contain have 1378:1381 arg1 findings 1369:1376 These findings 1363:1376 These findings 1363:1376 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 18 contain have 1378:1381 arg2 implications 1394:1405 structural implications 1383:1405 structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2) 1383:1495 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 1 19 theme virion 518:523 arg1 HA 556:557 HA 556:557 HA 556:557 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 19 theme virion 518:523 arg1 hemagglutinin 541:553 the virion surface protein hemagglutinin 514:553 the virion surface protein hemagglutinin (HA) 514:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 3 20 theme novel 759:763 arg1 pattern 765:771 a novel pattern 757:771 a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a 757:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 1 21 dep profile 473:479 arg1 the 448:450 the 448:450 the 448:450 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 2 22 theme changes 600:606 arg1 basis 585:589 the structural basis 570:589 the structural basis of these changes 570:606 However, the structural basis of these changes remains largely unclear. 28443077 1 23 theme surface 525:531 arg1 HA 556:557 HA 556:557 HA 556:557 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 23 theme surface 525:531 arg1 hemagglutinin 541:553 the virion surface protein hemagglutinin 514:553 the virion surface protein hemagglutinin (HA) 514:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 24 theme Structural 88:97 arg1 Basis 99:103 the Structural Basis 84:103 the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity 84:332 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 25 theme dynamics 679:686 arg1 simulations 688:698 molecular dynamics simulations 669:698 molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 669:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 26 theme influenza 217:225 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 26 theme influenza 217:225 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 27 theme structures 957:966 arg1 Comparison 926:935 Comparison 926:935 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations 926:1043 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 1 28 theme protein 533:539 arg1 HA 556:557 HA 556:557 HA 556:557 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 28 theme protein 533:539 arg1 hemagglutinin 541:553 the virion surface protein hemagglutinin 514:553 the virion surface protein hemagglutinin (HA) 514:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 29 theme Dynamics 10:17 arg1 Simulation 19:28 Molecular Dynamics Simulation 0:28 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer 0:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 30 gly glycosylated 707:718 arg1 trimers 723:729 the glycosylated HA trimers 703:729 the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 703:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 31 theme seasonal 208:215 arg1 influenza 217:225 seasonal influenza 208:225 seasonal influenza in humans since 1968 208:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 1 32 theme hemagglutinin 541:553 arg1 profile 473:479 N-glycosylation site profile 452:479 N-glycosylation site profile 452:479 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 32 theme hemagglutinin 541:553 arg1 activity 502:509 hemagglutination activity 485:509 hemagglutination activity 485:509 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 6 33 theme A 1489:1489 arg1 understanding 1415:1427 our understanding 1411:1427 our understanding of the phenotypic changes 1411:1453 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 33 theme A 1489:1489 arg1 evolution 1456:1464 evolution 1456:1464 evolution 1456:1464 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 33 theme A 1489:1489 arg1 fate 1471:1474 fate 1471:1474 fate 1471:1474 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 0 34 theme Molecular 0:8 arg1 Simulation 19:28 Molecular Dynamics Simulation 0:28 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer 0:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 35 theme trimers 723:729 arg1 simulations 688:698 molecular dynamics simulations 669:698 molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 669:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 36 theme human 314:318 arg1 immunity 325:332 the constantly changing human herd immunity 290:332 the constantly changing human herd immunity 290:332 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 37 theme glycosylated 975:986 arg1 trimers 991:997 the glycosylated HA trimers 971:997 the glycosylated HA trimers 971:997 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 1 38 gly N-glycosylation 452:466 arg1 HA 556:557 HA 556:557 HA 556:557 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 38 gly N-glycosylation 452:466 arg1 hemagglutinin 541:553 the virion surface protein hemagglutinin 514:553 the virion surface protein hemagglutinin (HA) 514:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 38 gly N-glycosylation 452:466 arg2 site 468:471 N-glycosylation site profile 452:479 N-glycosylation site profile 452:479 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 38 gly N-glycosylation 452:466 arg2 profile 473:479 N-glycosylation site profile 452:479 N-glycosylation site profile 452:479 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 39 theme Adaptive 109:116 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 39 theme Adaptive 109:116 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 40 theme glycan-shield 1152:1164 arg1 rearrangement 1166:1178 glycan-shield rearrangement 1152:1178 glycan-shield rearrangement 1152:1178 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 3 41 from unique 798:803 arg1 3C.2a 839:843 the new A(H3N2) epidemic clade 3C.2a 808:843 the new A(H3N2) epidemic clade 3C.2a 808:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 42 theme changing 305:312 arg1 immunity 325:332 the constantly changing human herd immunity 290:332 the constantly changing human herd immunity 290:332 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 6 43 theme structural 1383:1392 arg1 implications 1394:1405 structural implications 1383:1405 structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2) 1383:1495 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 3 44 theme glycosylated 707:718 arg1 trimers 723:729 the glycosylated HA trimers 703:729 the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 703:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 4 45 theme ligand 1133:1138 arg1 binding 1140:1146 the ligand binding 1129:1146 the ligand binding 1129:1146 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 6 46 theme influenza 1479:1487 arg1 H3N2 1491:1494 H3N2 1491:1494 H3N2 1491:1494 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 6 46 theme influenza 1479:1487 arg1 A 1489:1489 influenza A 1479:1489 influenza A(H3N2) 1479:1495 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 0 47 theme Influenza 37:45 arg1 H3N2 49:52 H3N2 49:52 H3N2 49:52 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 47 theme Influenza 37:45 arg1 A 47:47 Influenza A 37:47 the Influenza A(H3N2) Hemagglutinin Trimer 33:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 48 theme HA 720:721 arg1 trimers 723:729 the glycosylated HA trimers 703:729 the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 703:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 1 49 theme Increasing 335:344 arg1 evidence 346:353 Increasing evidence 335:353 Increasing evidence 335:353 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 3 50 theme epidemic 824:831 arg1 3C.2a 839:843 the new A(H3N2) epidemic clade 3C.2a 808:843 the new A(H3N2) epidemic clade 3C.2a 808:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 4 51 theme 3C.2a-specific 1020:1033 arg1 mutations 1035:1043 the 3C.2a-specific mutations 1016:1043 the 3C.2a-specific mutations 1016:1043 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 1 52 with concomitant 412:422 arg1 alterations 433:443 the alterations 429:443 the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA) 429:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 3 53 theme clade 833:837 arg1 3C.2a 839:843 the new A(H3N2) epidemic clade 3C.2a 808:843 the new A(H3N2) epidemic clade 3C.2a 808:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 1 54 theme N-glycosylation 452:466 arg1 profile 473:479 N-glycosylation site profile 452:479 N-glycosylation site profile 452:479 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 4 55 theme equilibrated 944:955 arg1 structures 957:966 the equilibrated structures 940:966 the equilibrated structures of the glycosylated HA trimers 940:997 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 0 56 theme Recent 135:140 arg1 A 174:174 the Recent Epidemic Clade 3C.2a. Influenza A 131:174 the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 131:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 56 theme Recent 135:140 arg1 H3N2 176:179 H3N2 176:179 H3N2 176:179 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 1 57 theme site 468:471 arg1 profile 473:479 N-glycosylation site profile 452:479 N-glycosylation site profile 452:479 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 58 theme A 47:47 arg1 Trimer 69:74 the Influenza A(H3N2) Hemagglutinin Trimer 33:74 the Influenza A(H3N2) Hemagglutinin Trimer 33:74 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 59 gly glycosylated 975:986 arg1 trimers 991:997 the glycosylated HA trimers 971:997 the glycosylated HA trimers 971:997 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 1 60 theme profile 473:479 arg1 alterations 433:443 the alterations 429:443 the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA) 429:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 61 from cause 199:203 arg1 humans 230:235 humans 230:235 humans since 1968 230:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 62 theme unique 798:803 arg1 pattern 765:771 a novel pattern 757:771 a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a 757:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 3 63 theme A 738:738 arg1 trimers 723:729 the glycosylated HA trimers 703:729 the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes 703:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 0 64 theme Clade 151:155 arg1 A 174:174 the Recent Epidemic Clade 3C.2a. Influenza A 131:174 the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 131:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 64 theme Clade 151:155 arg1 H3N2 176:179 H3N2 176:179 H3N2 176:179 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 5 65 theme binding 1342:1348 arg1 specificity 1350:1360 the ligand binding specificity 1331:1360 the ligand binding specificity 1331:1360 The results suggest that the 3C.2a strain has evolved an HA structure that is advantageous for evading pre-existing antibodies, while also increasing the ligand binding specificity. 28443077 3 66 theme attenuated 869:878 arg1 ability 880:886 attenuated ability 869:886 attenuated ability to agglutinate nonhuman erythrocytes 869:923 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 4 67 theme apical 1112:1117 arg1 space 1119:1123 the apical space 1108:1123 the apical space 1108:1123 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 0 68 theme Epidemic 142:149 arg1 A 174:174 the Recent Epidemic Clade 3C.2a. Influenza A 131:174 the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 131:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 68 theme Epidemic 142:149 arg1 H3N2 176:179 H3N2 176:179 H3N2 176:179 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 1 69 theme antigenic 373:381 arg1 change 383:388 the antigenic change 369:388 the antigenic change 369:388 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 1 70 theme hemagglutination 485:500 arg1 activity 502:509 hemagglutination activity 485:509 hemagglutination activity 485:509 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 3 71 theme Asn-X-Ser/Thr 776:788 arg1 sequons 790:796 Asn-X-Ser/Thr sequons 776:796 Asn-X-Ser/Thr sequons 776:796 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 4 72 theme drastic 1087:1093 arg1 reduction 1095:1103 a drastic reduction 1085:1103 a drastic reduction in the apical space for the ligand binding 1085:1146 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 3 73 contain has 753:755 arg1 A 738:738 the A 734:738 the A(H3N2) 734:744 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 3 73 contain has 753:755 arg2 pattern 765:771 a novel pattern 757:771 a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a 757:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 3 73 contain has 753:755 arg1 H3N2 740:743 H3N2 740:743 H3N2 740:743 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 1 74 theme activity 502:509 arg1 alterations 433:443 the alterations 429:443 the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA) 429:558 Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). 28443077 0 75 from humans 230:235 arg1 cause 199:203 a major cause 191:203 a major cause of seasonal influenza in humans since 1968 191:246 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 0 75 from humans 230:235 arg1 Evolution 118:126 Adaptive Evolution 109:126 Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) 109:180 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 3 76 theme sequons 790:796 arg1 pattern 765:771 a novel pattern 757:771 a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a 757:843 To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C.2a and is characterized by attenuated ability to agglutinate nonhuman erythrocytes. 28443077 6 77 theme phenotypic 1436:1445 arg1 changes 1447:1453 the phenotypic changes 1432:1453 the phenotypic changes 1432:1453 These findings have structural implications for our understanding of the phenotypic changes, evolution, and fate of influenza A(H3N2). 28443077 4 78 from reduction 1095:1103 arg1 space 1119:1123 the apical space 1108:1123 the apical space 1108:1123 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 28443077 0 79 theme antigenic 268:276 arg1 drift 278:282 antigenic drift 268:282 antigenic drift 268:282 Molecular Dynamics Simulation of the Influenza A(H3N2) Hemagglutinin Trimer Reveals the Structural Basis for Adaptive Evolution of the Recent Epidemic Clade 3C.2a. Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. 28443077 4 80 theme trimers 991:997 arg1 structures 957:966 the equilibrated structures 940:966 the equilibrated structures of the glycosylated HA trimers 940:997 Comparison of the equilibrated structures of the glycosylated HA trimers with and without the 3C.2a-specific mutations reveals that the mutations could induce a drastic reduction in the apical space for the ligand binding via glycan-shield rearrangement. 26821880 6 0 used used 947:950 arg2 simulation 931:940 molecular dynamics simulation 912:940 molecular dynamics simulation 912:940 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 6 0 used used 947:950 arg2 substrates 896:905 docking the donor and acceptor substrates 865:905 docking the donor and acceptor substrates 865:905 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 6 0 used used 947:950 arg2 methods 856:862 Various alignment methods 838:862 Various alignment methods 838:862 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 6 1 theme homology 971:978 arg1 models 980:985 seven homology models 965:985 seven homology models of GnT-V 965:994 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 1 2 link β-1-6-linked 318:329 arg1 branching 331:339 the β-1-6-linked branching 314:339 the β-1-6-linked branching of N-linked oligosaccharides 314:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 3 theme GlcNAc 191:196 arg1 transfer 179:186 the transfer 175:186 the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 175:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 8 4 theme enzyme 1166:1171 arg1 site 1183:1186 the enzyme catalytic site 1162:1186 the enzyme catalytic site 1162:1186 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 4 5 theme information 641:651 arg1 lack 633:636 the lack 629:636 the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates 629:753 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 5 6 theme homology 819:826 arg1 modeling 828:835 homology modeling 819:835 homology modeling 819:835 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 4 7 theme specific 585:592 arg1 inhibitors 594:603 potent and specific inhibitors 574:603 potent and specific inhibitors of GnT-V 574:612 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 8 8 dep donor 1229:1233 arg1 the 1225:1227 the 1225:1227 the 1225:1227 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 2 9 theme cancer 423:428 arg1 growth 430:435 cancer growth 423:435 cancer growth 423:435 β-1-6-GlcNAc-branched N-glycans are associated with cancer growth and metastasis. 26821880 8 10 from useful 1255:1260 arg1 studies 1265:1271 studies 1265:1271 studies of the catalytic mechanism 1265:1298 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 10 from useful 1255:1260 arg1 design 1304:1309 design 1304:1309 design of inhibitors of GnT-V 1304:1332 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 1 11 theme glycoproteins 289:301 arg1 structure 276:284 the trimannosyl core structure 255:284 the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 255:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 8 12 theme binding 1192:1198 arg1 information 1200:1210 binding information 1192:1210 binding information obtained for the donor and acceptor 1192:1246 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 6 13 theme docking 865:871 arg1 substrates 896:905 docking the donor and acceptor substrates 865:905 docking the donor and acceptor substrates 865:905 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 1 14 theme enzyme 71:76 arg1 UDP-N-acetylglucosamine 78:100 The enzyme UDP-N-acetylglucosamine 67:100 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 67:369 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 2 15 theme β-1-6-GlcNAc-branched 371:391 arg1 N-glycans 393:401 β-1-6-GlcNAc-branched N-glycans 371:401 β-1-6-GlcNAc-branched N-glycans 371:401 β-1-6-GlcNAc-branched N-glycans are associated with cancer growth and metastasis. 26821880 1 16 link α-1-6-linked 231:242 arg1 mannose 244:250 the α-1-6-linked mannose 227:250 the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 227:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 6 17 theme substrates 1032:1041 arg1 binding 1017:1023 the binding 1013:1023 the binding of its substrates 1013:1041 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 5 18 theme GnT-V 798:802 arg1 result 809:814 a result 807:814 a result of homology modeling 807:835 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 5 18 theme GnT-V 798:802 arg1 structure 785:793 the first 3D structure 772:793 the first 3D structure of GnT-V 772:802 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 4 19 theme enzyme 695:700 arg1 structure 678:686 the three-dimensional structure 656:686 the three-dimensional structure of the enzyme 656:700 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 8 20 theme mechanism 1290:1298 arg1 studies 1265:1271 studies 1265:1271 studies of the catalytic mechanism 1265:1298 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 20 theme mechanism 1290:1298 arg1 design 1304:1309 design 1304:1309 design of inhibitors of GnT-V 1304:1332 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 7 21 with consistent 1071:1080 arg1 data 1110:1113 available experimental data 1087:1113 available experimental data 1087:1113 The best homology model is consistent with available experimental data. 26821880 8 22 from studies 1265:1271 arg1 structure 1149:1157 the structure 1145:1157 the structure of the enzyme catalytic site 1145:1186 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 22 from studies 1265:1271 arg1 useful 1255:1260 useful 1255:1260 useful 1255:1260 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 22 from studies 1265:1271 arg1 model 1138:1142 The three-dimensional model 1116:1142 The three-dimensional model 1116:1142 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 3 23 theme GnT-V 482:486 arg1 inhibition 468:477 the inhibition 464:477 the inhibition of GnT-V 464:486 Therefore, the inhibition of GnT-V represents a key target for anti-cancer drug development. 26821880 1 24 gly glycoproteins 289:301 arg1 glycoproteins 289:301 glycoproteins 289:301 glycoproteins 289:301 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 25 theme α-d-mannoside 103:115 arg1 GnT-V 158:162 GnT-V 158:162 GnT-V 158:162 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 25 theme α-d-mannoside 103:115 arg1 V 155:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V 103:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) 103:163 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 26 theme UDP-GlcNAc 207:216 arg1 donor 218:222 the UDP-GlcNAc donor 203:222 the UDP-GlcNAc donor 203:222 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 0 27 theme homology 18:25 arg1 model 27:31 Three-dimensional homology model 0:31 Three-dimensional homology model of GlcNAc-TV glycosyltransferase 0:64 Three-dimensional homology model of GlcNAc-TV glycosyltransferase. 26821880 6 28 theme Various 838:844 arg1 methods 856:862 Various alignment methods 838:862 Various alignment methods 838:862 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 1 29 theme β-1-6 117:121 arg1 GnT-V 158:162 GnT-V 158:162 GnT-V 158:162 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 29 theme β-1-6 117:121 arg1 V 155:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V 103:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) 103:163 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 0 30 theme Three-dimensional 0:16 arg1 model 27:31 Three-dimensional homology model 0:31 Three-dimensional homology model of GlcNAc-TV glycosyltransferase 0:64 Three-dimensional homology model of GlcNAc-TV glycosyltransferase. 26821880 8 31 theme three-dimensional 1120:1136 arg1 useful 1255:1260 useful 1255:1260 useful 1255:1260 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 31 theme three-dimensional 1120:1136 arg1 model 1138:1142 The three-dimensional model 1116:1142 The three-dimensional model 1116:1142 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 1 32 theme N-acetylglucosaminyltransferase 123:153 arg1 GnT-V 158:162 GnT-V 158:162 GnT-V 158:162 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 32 theme N-acetylglucosaminyltransferase 123:153 arg1 V 155:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V 103:155 α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) 103:163 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 3 33 theme key 501:503 arg1 target 505:510 a key target 499:510 a key target for anti-cancer drug development 499:543 Therefore, the inhibition of GnT-V represents a key target for anti-cancer drug development. 26821880 1 34 theme β-1-6-linked 318:329 arg1 branching 331:339 the β-1-6-linked branching 314:339 the β-1-6-linked branching of N-linked oligosaccharides 314:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 8 35 theme inhibitors 1314:1323 arg1 studies 1265:1271 studies 1265:1271 studies of the catalytic mechanism 1265:1298 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 35 theme inhibitors 1314:1323 arg1 design 1304:1309 design 1304:1309 design of inhibitors of GnT-V 1304:1332 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 7 36 theme homology 1053:1060 arg1 model 1062:1066 The best homology model 1044:1066 The best homology model 1044:1066 The best homology model is consistent with available experimental data. 26821880 7 36 theme homology 1053:1060 arg1 consistent 1071:1080 consistent 1071:1080 consistent 1071:1080 The best homology model is consistent with available experimental data. 26821880 6 37 theme dynamics 922:929 arg1 simulation 931:940 molecular dynamics simulation 912:940 molecular dynamics simulation 912:940 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 7 38 theme available 1087:1095 arg1 data 1110:1113 available experimental data 1087:1113 available experimental data 1087:1113 The best homology model is consistent with available experimental data. 26821880 1 39 theme α-1-6-linked 231:242 arg1 mannose 244:250 the α-1-6-linked mannose 227:250 the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 227:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 0 40 theme glycosyltransferase 46:64 arg1 model 27:31 Three-dimensional homology model 0:31 Three-dimensional homology model of GlcNAc-TV glycosyltransferase 0:64 Three-dimensional homology model of GlcNAc-TV glycosyltransferase. 26821880 5 41 theme first 776:780 arg1 result 809:814 a result 807:814 a result of homology modeling 807:835 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 5 41 theme first 776:780 arg1 structure 785:793 the first 3D structure 772:793 the first 3D structure of GnT-V 772:802 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 6 42 theme molecular 912:920 arg1 simulation 931:940 molecular dynamics simulation 912:940 molecular dynamics simulation 912:940 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 3 43 theme anti-cancer 516:526 arg1 development 533:543 anti-cancer drug development 516:543 anti-cancer drug development 516:543 Therefore, the inhibition of GnT-V represents a key target for anti-cancer drug development. 26821880 0 44 theme GlcNAc-TV 36:44 arg1 glycosyltransferase 46:64 GlcNAc-TV glycosyltransferase 36:64 GlcNAc-TV glycosyltransferase 36:64 Three-dimensional homology model of GlcNAc-TV glycosyltransferase. 26821880 1 45 theme N-linked 344:351 arg1 oligosaccharides 353:368 N-linked oligosaccharides 344:368 N-linked oligosaccharides 344:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 5 46 theme 3D 782:783 arg1 result 809:814 a result 807:814 a result of homology modeling 807:835 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 5 46 theme 3D 782:783 arg1 structure 785:793 the first 3D structure 772:793 the first 3D structure of GnT-V 772:802 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 8 47 theme GnT-V 1328:1332 arg1 inhibitors 1314:1323 inhibitors 1314:1323 inhibitors of GnT-V 1314:1332 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 48 from design 1304:1309 arg1 structure 1149:1157 the structure 1145:1157 the structure of the enzyme catalytic site 1145:1186 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 48 from design 1304:1309 arg1 useful 1255:1260 useful 1255:1260 useful 1255:1260 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 48 from design 1304:1309 arg1 model 1138:1142 The three-dimensional model 1116:1142 The three-dimensional model 1116:1142 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 3 49 theme drug 528:531 arg1 development 533:543 anti-cancer drug development 516:543 anti-cancer drug development 516:543 Therefore, the inhibition of GnT-V represents a key target for anti-cancer drug development. 26821880 1 50 theme oligosaccharides 353:368 arg1 branching 331:339 the β-1-6-linked branching 314:339 the β-1-6-linked branching of N-linked oligosaccharides 314:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 7 51 theme experimental 1097:1108 arg1 data 1110:1113 available experimental data 1087:1113 available experimental data 1087:1113 The best homology model is consistent with available experimental data. 26821880 6 52 dep donor 877:881 arg1 the 873:875 the 873:875 the 873:875 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 4 53 theme three-dimensional 660:676 arg1 structure 678:686 the three-dimensional structure 656:686 the three-dimensional structure of the enzyme 656:700 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 6 54 theme GnT-V 990:994 arg1 models 980:985 seven homology models 965:985 seven homology models of GnT-V 965:994 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 4 55 from lack 633:636 arg1 structure 678:686 the three-dimensional structure 656:686 the three-dimensional structure of the enzyme 656:700 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 4 55 from lack 633:636 arg1 characteristics 721:735 the binding characteristics 709:735 the binding characteristics of its substrates 709:753 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 6 56 theme acceptor 887:894 arg1 substrates 896:905 docking the donor and acceptor substrates 865:905 docking the donor and acceptor substrates 865:905 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 1 57 link N-linked 344:351 arg1 oligosaccharides 353:368 N-linked oligosaccharides 344:368 N-linked oligosaccharides 344:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 4 58 theme GnT-V 608:612 arg1 inhibitors 594:603 potent and specific inhibitors 574:603 potent and specific inhibitors of GnT-V 574:612 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 8 59 theme site 1183:1186 arg1 information 1200:1210 binding information 1192:1210 binding information obtained for the donor and acceptor 1192:1246 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 59 theme site 1183:1186 arg1 structure 1149:1157 the structure 1145:1157 the structure of the enzyme catalytic site 1145:1186 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 59 theme site 1183:1186 arg1 useful 1255:1260 useful 1255:1260 useful 1255:1260 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 8 59 theme site 1183:1186 arg1 model 1138:1142 The three-dimensional model 1116:1142 The three-dimensional model 1116:1142 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 6 60 theme alignment 846:854 arg1 methods 856:862 Various alignment methods 838:862 Various alignment methods 838:862 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 8 61 theme catalytic 1173:1181 arg1 site 1183:1186 the enzyme catalytic site 1162:1186 the enzyme catalytic site 1162:1186 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 5 62 theme modeling 828:835 arg1 result 809:814 a result 807:814 a result of homology modeling 807:835 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 5 62 theme modeling 828:835 arg1 structure 785:793 the first 3D structure 772:793 the first 3D structure of GnT-V 772:802 Here we present the first 3D structure of GnT-V as a result of homology modeling. 26821880 8 63 theme catalytic 1280:1288 arg1 mechanism 1290:1298 the catalytic mechanism 1276:1298 the catalytic mechanism 1276:1298 The three-dimensional model, the structure of the enzyme catalytic site and binding information obtained for the donor and acceptor can be useful in studies of the catalytic mechanism and design of inhibitors of GnT-V. 26821880 1 64 from donor 218:222 arg1 transfer 179:186 the transfer 175:186 the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 175:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 65 dep UDP-N-acetylglucosamine 78:100 arg1 catalyzes 165:173 catalyzes 165:173 catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 165:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 1 66 theme trimannosyl 259:269 arg1 structure 276:284 the trimannosyl core structure 255:284 the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 255:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 4 67 theme inhibitors 594:603 arg1 development 559:569 the development 555:569 the development of potent and specific inhibitors of GnT-V 555:612 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 6 68 theme donor 877:881 arg1 substrates 896:905 docking the donor and acceptor substrates 865:905 docking the donor and acceptor substrates 865:905 Various alignment methods, docking the donor and acceptor substrates, and molecular dynamics simulation were used to construct seven homology models of GnT-V and characterize the binding of its substrates. 26821880 7 69 theme best 1048:1051 arg1 model 1062:1066 The best homology model 1044:1066 The best homology model 1044:1066 The best homology model is consistent with available experimental data. 26821880 7 69 theme best 1048:1051 arg1 consistent 1071:1080 consistent 1071:1080 consistent 1071:1080 The best homology model is consistent with available experimental data. 26821880 4 70 theme binding 713:719 arg1 characteristics 721:735 the binding characteristics 709:735 the binding characteristics of its substrates 709:753 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 1 71 theme core 271:274 arg1 structure 276:284 the trimannosyl core structure 255:284 the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 255:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26821880 4 72 theme potent 574:579 arg1 inhibitors 594:603 potent and specific inhibitors 574:603 potent and specific inhibitors of GnT-V 574:612 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 4 73 theme substrates 744:753 arg1 characteristics 721:735 the binding characteristics 709:735 the binding characteristics of its substrates 709:753 However, the development of potent and specific inhibitors of GnT-V is hampered by the lack of information on the three-dimensional structure of the enzyme and on the binding characteristics of its substrates. 26821880 1 74 theme structure 276:284 arg1 mannose 244:250 the α-1-6-linked mannose 227:250 the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides 227:368 The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. 26578555 3 0 theme data 304:307 arg1 components 362:371 three inseparable components 344:371 three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 344:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 0 theme data 304:307 arg1 set 337:339 a set 335:339 a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 335:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 0 theme data 304:307 arg1 element 309:315 The core data element 295:315 The core data element of SugarBindDB 295:330 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 5 1 theme visualisation 590:602 arg1 tools 604:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 1 2 theme pathogen 152:159 arg1 lectins 161:167 human pathogen lectins 146:167 human pathogen lectins 146:167 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 7 3 theme substructure 835:846 arg1 tool 855:858 a substructure search tool 833:858 a substructure search tool that maps each ligand to full structures where it occurs 833:915 It is tightly bound to the latter via a substructure search tool that maps each ligand to full structures where it occurs. 26578555 9 4 from http 1217:1220 arg1 accessible 1202:1211 accessible 1202:1211 accessible 1202:1211 SugarBindDB is accessible at: http://sugarbind.expasy.org. 26578555 8 5 theme SugarBindDB 956:966 arg1 pair 948:951 a glycan-lectin binding pair 924:951 a glycan-lectin binding pair of SugarBindDB 924:966 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 1 6 theme lectins 161:167 arg1 binding 135:141 glycan binding 128:141 glycan binding of human pathogen lectins and adhesins 128:180 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 5 7 theme pathogen 675:682 arg1 binding 684:690 pathogen binding 675:690 pathogen binding 675:690 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 7 8 theme full 885:888 arg1 structures 890:899 full structures 885:899 full structures where it occurs 885:915 It is tightly bound to the latter via a substructure search tool that maps each ligand to full structures where it occurs. 26578555 3 9 dep components 362:371 arg1 ligand 426:431 a glycan ligand 417:431 a glycan ligand 417:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 9 dep components 362:371 arg1 agent 389:393 the pathogenic agent 374:393 the pathogenic agent 374:393 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 9 dep components 362:371 arg1 lectin/adhesin 398:411 a lectin/adhesin 396:411 a lectin/adhesin 396:411 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 2 10 theme published 274:282 arg1 reference 284:292 at least one published reference 261:292 at least one published reference 261:292 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 5 11 from role 656:659 arg1 binding 684:690 pathogen binding 675:690 pathogen binding 675:690 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 8 12 theme substructure 1097:1108 arg1 search 1110:1115 substructure search 1097:1115 substructure search 1097:1115 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 9 13 dep http 1217:1220 arg1 //sugarbind.expasy.org 1222:1243 //sugarbind.expasy.org 1222:1243 http://sugarbind.expasy.org 1217:1243 SugarBindDB is accessible at: http://sugarbind.expasy.org. 26578555 8 14 theme glycan-lectin 926:938 arg1 pair 948:951 a glycan-lectin binding pair 924:951 a glycan-lectin binding pair of SugarBindDB 924:966 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 5 15 theme search 567:572 arg1 tools 604:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 5 16 theme Several 559:565 arg1 tools 604:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 1 17 theme SugarBind 75:83 arg1 SugarBindDB 95:105 SugarBindDB 95:105 SugarBindDB 95:105 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 1 17 theme SugarBind 75:83 arg1 Database 85:92 The SugarBind Database 71:92 The SugarBind Database (SugarBindDB) 71:106 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 0 18 theme host-pathogen 43:55 arg1 interactions 57:68 glycan-mediated host-pathogen interactions 27:68 glycan-mediated host-pathogen interactions 27:68 SugarBindDB, a resource of glycan-mediated host-pathogen interactions. 26578555 8 19 theme binding 940:946 arg1 pair 948:951 a glycan-lectin binding pair 924:951 a glycan-lectin binding pair of SugarBindDB 924:966 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 1 20 theme adhesins 173:180 arg1 binding 135:141 glycan binding 128:141 glycan binding of human pathogen lectins and adhesins 128:180 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 3 21 theme components 362:371 arg1 components 362:371 three inseparable components 344:371 three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 344:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 21 theme components 362:371 arg1 set 337:339 a set 335:339 a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 335:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 21 theme components 362:371 arg1 element 309:315 The core data element 295:315 The core data element of SugarBindDB 295:330 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 4 22 dep entity 439:444 arg1 agent 447:451 agent 447:451 agent 447:451 Each entity (agent, lectin or ligand) is described by a range of properties that are summarized in an entity-dedicated page. 26578555 4 22 dep entity 439:444 arg1 ligand 464:469 ligand 464:469 ligand 464:469 Each entity (agent, lectin or ligand) is described by a range of properties that are summarized in an entity-dedicated page. 26578555 4 22 dep entity 439:444 arg1 lectin 454:459 lectin 454:459 lectin 454:459 Each entity (agent, lectin or ligand) is described by a range of properties that are summarized in an entity-dedicated page. 26578555 5 23 theme navigation 575:584 arg1 tools 604:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools 559:608 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 6 24 theme glycan-relaled 737:750 arg1 UniCarbKB 784:792 UniCarbKB 784:792 UniCarbKB 784:792 The database is cross-linked to protein and glycan-relaled resources such as UniProtKB and UniCarbKB. 26578555 6 24 theme glycan-relaled 737:750 arg1 resources 752:760 glycan-relaled resources 737:760 glycan-relaled resources such as UniProtKB and UniCarbKB 737:792 The database is cross-linked to protein and glycan-relaled resources such as UniProtKB and UniCarbKB. 26578555 6 24 theme glycan-relaled 737:750 arg1 UniProtKB 770:778 UniProtKB 770:778 UniProtKB 770:778 The database is cross-linked to protein and glycan-relaled resources such as UniProtKB and UniCarbKB. 26578555 8 25 theme glycosylation 1152:1164 arg1 search 1110:1115 substructure search 1097:1115 substructure search 1097:1115 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 8 25 theme glycosylation 1152:1164 arg1 knowledge 1125:1133 the knowledge 1121:1133 the knowledge of site-specific glycosylation stored in UniCarbKB 1121:1184 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 8 26 theme protein-protein 1020:1034 arg1 interaction 1036:1046 a glycan-mediated protein-protein interaction 1002:1046 a glycan-mediated protein-protein interaction 1002:1046 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 8 26 theme protein-protein 1020:1034 arg1 interaction 1080:1090 a lectin-glycoprotein interaction 1058:1090 a lectin-glycoprotein interaction 1058:1090 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 3 27 theme inseparable 350:360 arg1 components 362:371 three inseparable components 344:371 three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 344:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 8 28 theme glycan-mediated 1004:1018 arg1 interaction 1036:1046 a glycan-mediated protein-protein interaction 1002:1046 a glycan-mediated protein-protein interaction 1002:1046 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 8 28 theme glycan-mediated 1004:1018 arg1 interaction 1080:1090 a lectin-glycoprotein interaction 1058:1090 a lectin-glycoprotein interaction 1058:1090 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 0 29 theme glycan-mediated 27:41 arg1 interactions 57:68 glycan-mediated host-pathogen interactions 27:68 glycan-mediated host-pathogen interactions 27:68 SugarBindDB, a resource of glycan-mediated host-pathogen interactions. 26578555 4 30 theme properties 499:508 arg1 range 490:494 a range 488:494 a range of properties that are summarized in an entity-dedicated page 488:556 Each entity (agent, lectin or ligand) is described by a range of properties that are summarized in an entity-dedicated page. 26578555 5 31 theme glycans 664:670 arg1 role 656:659 the functional role 641:659 the functional role of glycans in pathogen binding 641:690 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 2 32 theme binding 229:235 arg1 pair 237:240 each glycan-protein binding pair 209:240 each glycan-protein binding pair 209:240 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 3 33 theme glycan 419:424 arg1 ligand 426:431 a glycan ligand 417:431 a glycan ligand 417:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 33 theme glycan 419:424 arg1 agent 389:393 the pathogenic agent 374:393 the pathogenic agent 374:393 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 8 34 theme interaction 1036:1046 arg1 identification 984:997 the identification 980:997 the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB 980:1184 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 2 35 theme glycan-protein 214:227 arg1 pair 237:240 each glycan-protein binding pair 209:240 each glycan-protein binding pair 209:240 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 8 36 gly lectin-glycoprotein 1060:1078 arg1 lectin-glycoprotein 1060:1078 lectin-glycoprotein 1060:1078 lectin-glycoprotein 1060:1078 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 0 37 theme interactions 57:68 arg1 SugarBindDB 0:10 SugarBindDB 0:10 SugarBindDB 0:10 SugarBindDB, a resource of glycan-mediated host-pathogen interactions. 26578555 0 37 theme interactions 57:68 arg1 resource 15:22 a resource 13:22 a resource of glycan-mediated host-pathogen interactions 13:68 SugarBindDB, a resource of glycan-mediated host-pathogen interactions. 26578555 8 38 theme lectin-glycoprotein 1060:1078 arg1 interaction 1036:1046 a glycan-mediated protein-protein interaction 1002:1046 a glycan-mediated protein-protein interaction 1002:1046 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 8 38 theme lectin-glycoprotein 1060:1078 arg1 interaction 1080:1090 a lectin-glycoprotein interaction 1058:1090 a lectin-glycoprotein interaction 1058:1090 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 3 39 theme pathogenic 378:387 arg1 ligand 426:431 a glycan ligand 417:431 a glycan ligand 417:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 39 theme pathogenic 378:387 arg1 agent 389:393 the pathogenic agent 374:393 the pathogenic agent 374:393 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 39 theme pathogenic 378:387 arg1 lectin/adhesin 398:411 a lectin/adhesin 396:411 a lectin/adhesin 396:411 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 40 theme core 299:302 arg1 components 362:371 three inseparable components 344:371 three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 344:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 40 theme core 299:302 arg1 set 337:339 a set 335:339 a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 335:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 40 theme core 299:302 arg1 element 309:315 The core data element 295:315 The core data element of SugarBindDB 295:330 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 2 41 theme curated 191:197 arg1 It 183:184 It 183:184 It 183:184 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 2 41 theme curated 191:197 arg1 database 199:206 a curated database 189:206 a curated database; each glycan-protein binding pair is associated with at least one published reference 189:292 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 1 42 theme glycan 128:133 arg1 binding 135:141 glycan binding 128:141 glycan binding of human pathogen lectins and adhesins 128:180 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 5 43 theme functional 645:654 arg1 role 656:659 the functional role 641:659 the functional role of glycans in pathogen binding 641:690 Several search, navigation and visualisation tools are implemented to investigate the functional role of glycans in pathogen binding. 26578555 1 44 theme binding 135:141 arg1 knowledge 115:123 knowledge 115:123 knowledge of glycan binding of human pathogen lectins and adhesins 115:180 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 26578555 7 45 theme search 848:853 arg1 tool 855:858 a substructure search tool 833:858 a substructure search tool that maps each ligand to full structures where it occurs 833:915 It is tightly bound to the latter via a substructure search tool that maps each ligand to full structures where it occurs. 26578555 4 46 theme entity-dedicated 536:551 arg1 page 553:556 an entity-dedicated page 533:556 an entity-dedicated page 533:556 Each entity (agent, lectin or ligand) is described by a range of properties that are summarized in an entity-dedicated page. 26578555 2 47 dep database 199:206 arg1 associated 245:254 associated 245:254 is associated with at least one published reference 242:292 It is a curated database; each glycan-protein binding pair is associated with at least one published reference. 26578555 3 48 theme SugarBindDB 320:330 arg1 components 362:371 three inseparable components 344:371 three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 344:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 48 theme SugarBindDB 320:330 arg1 set 337:339 a set 335:339 a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand 335:431 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 3 48 theme SugarBindDB 320:330 arg1 element 309:315 The core data element 295:315 The core data element of SugarBindDB 295:330 The core data element of SugarBindDB is a set of three inseparable components: the pathogenic agent, a lectin/adhesin and a glycan ligand. 26578555 8 49 theme site-specific 1138:1150 arg1 glycosylation 1152:1164 site-specific glycosylation 1138:1164 site-specific glycosylation stored in UniCarbKB 1138:1184 Thus, a glycan-lectin binding pair of SugarBindDB can lead to the identification of a glycan-mediated protein-protein interaction, that is, a lectin-glycoprotein interaction, via substructure search and the knowledge of site-specific glycosylation stored in UniCarbKB. 26578555 1 50 theme human 146:150 arg1 lectins 161:167 human pathogen lectins 146:167 human pathogen lectins 146:167 The SugarBind Database (SugarBindDB) covers knowledge of glycan binding of human pathogen lectins and adhesins. 24308457 3 0 theme extracellular 696:708 arg1 terminal 710:717 the extracellular terminal 692:717 the extracellular terminal of the protein of interest 692:744 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 5 1 theme receptor 1220:1227 arg1 I. 1234:1235 receptor type I. 1220:1235 receptor type I. 1220:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 6 2 dep functions 1438:1446 arg1 the 1434:1436 the 1434:1436 the 1434:1436 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 1 3 theme imaging 331:337 arg1 glycosylation 339:351 imaging glycosylation 331:351 imaging glycosylation of a specific protein of interest (POI) 331:391 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 3 4 theme FRET 645:648 arg1 acceptor 650:657 a FRET acceptor 643:657 a FRET acceptor 643:657 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 5 5 theme type 1229:1232 arg1 I. 1234:1235 receptor type I. 1220:1235 receptor type I. 1220:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 2 6 theme live 517:520 arg1 cells 522:526 live cells 517:526 live cells 517:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 5 7 theme epidermal 1150:1158 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 3 8 theme glycan 550:555 arg1 labeling 557:564 metabolic glycan labeling 540:564 metabolic glycan labeling 540:564 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 5 9 theme cell 1102:1105 arg1 receptors 1115:1123 several important cell surface receptors 1084:1123 several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1084:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 9 theme cell 1102:1105 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 9 theme cell 1102:1105 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 9 theme cell 1102:1105 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 4 10 theme intermolecular 871:884 arg1 FRET 886:889 the intermolecular FRET 867:889 the intermolecular FRET 867:889 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 2 11 theme glycans 506:512 arg1 imaging 495:501 protein-specific imaging 478:501 protein-specific imaging of glycans on live cells 478:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 3 12 theme protein 726:732 arg1 acceptor 650:657 a FRET acceptor 643:657 a FRET acceptor 643:657 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 3 12 theme protein 726:732 arg1 donor 665:669 a donor 663:669 a donor 663:669 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 3 12 theme protein 726:732 arg1 terminal 710:717 the extracellular terminal 692:717 the extracellular terminal of the protein of interest 692:744 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 4 13 theme effective 843:851 arg1 FRET 853:856 effective FRET 843:856 effective FRET 843:856 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 4 14 theme donor-acceptor 780:793 arg1 distance 795:802 The intramolecular donor-acceptor distance 761:802 The intramolecular donor-acceptor distance for the POI 761:814 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 6 15 theme proteins 1485:1492 arg1 dynamics 1452:1459 dynamics 1452:1459 dynamics 1452:1459 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 6 15 theme proteins 1485:1492 arg1 functions 1438:1446 functions 1438:1446 functions 1438:1446 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 5 16 theme important 1092:1100 arg1 receptors 1115:1123 several important cell surface receptors 1084:1123 several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1084:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 16 theme important 1092:1100 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 16 theme important 1092:1100 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 16 theme important 1092:1100 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 17 theme specific 358:365 arg1 protein 367:373 a specific protein 356:373 a specific protein of interest (POI) 356:391 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 0 18 theme protein-specific 37:52 arg1 imaging 54:60 protein-specific imaging 37:60 protein-specific imaging of cell-surface glycans 37:84 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 24308457 5 19 dep visualizing 1053:1063 arg1 revealed 1274:1281 revealed 1274:1281 revealed that the sialylation might be important for β2 integrin activation 1274:1348 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 20 theme Metabolic 87:95 arg1 tool 223:226 a powerful tool 212:226 a powerful tool for imaging glycome 212:246 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 1 20 theme Metabolic 87:95 arg1 labeling 97:104 Metabolic labeling 87:104 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry 87:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 1 21 gly glycosylation 339:351 arg1 protein 367:373 a specific protein 356:373 a specific protein of interest (POI) 356:391 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 6 22 theme live-cell 1385:1393 arg1 studies 1395:1401 the live-cell studies 1381:1401 the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins 1381:1492 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 0 23 theme cis-membrane 2:13 arg1 method 26:31 A cis-membrane FRET-based method 0:31 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 0:85 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 24308457 1 24 theme protein 367:373 arg1 glycosylation 339:351 imaging glycosylation 331:351 imaging glycosylation of a specific protein of interest (POI) 331:391 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 5 25 theme surface 1107:1113 arg1 receptors 1115:1123 several important cell surface receptors 1084:1123 several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1084:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 25 theme surface 1107:1113 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 25 theme surface 1107:1113 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 25 theme surface 1107:1113 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 26 dep αXβ2 1144:1147 arg1 I. 1234:1235 receptor type I. 1220:1235 receptor type I. 1220:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 27 theme method 1043:1048 arg1 capability 998:1007 the capability 994:1007 the capability of this cis-membrane FRET imaging method 994:1048 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 28 theme β2 1327:1328 arg1 activation 1339:1348 β2 integrin activation 1327:1348 β2 integrin activation 1327:1348 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 29 with labeling 97:104 arg1 reproters 131:139 chemical reproters 122:139 chemical reproters (e.g., alkyne or azide) 122:163 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 5 30 theme transforming 1188:1199 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 31 theme glycans 109:115 arg1 tool 223:226 a powerful tool 212:226 a powerful tool for imaging glycome 212:246 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 1 31 theme glycans 109:115 arg1 labeling 97:104 Metabolic labeling 87:104 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry 87:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 5 32 theme integrin 1330:1337 arg1 activation 1339:1348 β2 integrin activation 1327:1348 β2 integrin activation 1327:1348 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 33 theme bioorthogonal 185:197 arg1 chemistry 199:207 bioorthogonal chemistry 185:207 bioorthogonal chemistry 185:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 0 34 theme FRET-based 15:24 arg1 method 26:31 A cis-membrane FRET-based method 0:31 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 0:85 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 24308457 5 35 theme growth 1201:1206 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 4 36 theme excess 915:920 arg1 distant 954:960 distant 954:960 distant 954:960 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 4 36 theme excess 915:920 arg1 acceptors 922:930 the excess acceptors 911:930 the excess acceptors on other proteins 911:948 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 1 37 dep alkyne 148:153 arg1 e.g. 142:145 e.g. 142:145 e.g. 142:145 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 2 38 theme methodology 454:464 arg1 development 413:423 the development 409:423 the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells 409:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 3 39 theme interest 737:744 arg1 protein 726:732 the protein 722:732 the protein of interest 722:744 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 4 40 theme other 935:939 arg1 proteins 941:948 other proteins 935:948 other proteins 935:948 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 1 41 theme interest 378:385 arg1 protein 367:373 a specific protein 356:373 a specific protein of interest (POI) 356:391 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 3 42 theme site-specific 586:598 arg1 labeling 608:615 site-specific protein labeling 586:615 site-specific protein labeling 586:615 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 2 43 theme FRET-based 443:452 arg1 methodology 454:464 a cis-membrane FRET-based methodology 428:464 a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells 428:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 5 44 theme growth 1160:1165 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 2 45 from imaging 495:501 arg1 cells 522:526 live cells 517:526 live cells 517:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 3 46 theme protein 600:606 arg1 labeling 608:615 site-specific protein labeling 586:615 site-specific protein labeling 586:615 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 2 47 theme cis-membrane 430:441 arg1 methodology 454:464 a cis-membrane FRET-based methodology 428:464 a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells 428:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 3 48 with conjunction 569:579 arg1 labeling 608:615 site-specific protein labeling 586:615 site-specific protein labeling 586:615 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 1 49 with conjunction 168:178 arg1 chemistry 199:207 bioorthogonal chemistry 185:207 bioorthogonal chemistry 185:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 2 50 theme protein-specific 478:493 arg1 imaging 495:501 protein-specific imaging 478:501 protein-specific imaging of glycans on live cells 478:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 5 51 theme FRET 1030:1033 arg1 method 1043:1048 this cis-membrane FRET imaging method 1012:1048 this cis-membrane FRET imaging method 1012:1048 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 52 theme receptors 1115:1123 arg1 sialylation 1069:1079 the sialylation 1065:1079 the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1065:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 53 theme chemical 122:129 arg1 reproters 131:139 chemical reproters 122:139 chemical reproters (e.g., alkyne or azide) 122:163 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 2 54 from glycans 506:512 arg1 cells 522:526 live cells 517:526 live cells 517:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 5 55 theme imaging 1035:1041 arg1 method 1043:1048 this cis-membrane FRET imaging method 1012:1048 this cis-membrane FRET imaging method 1012:1048 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 4 56 from acceptors 922:930 arg1 proteins 941:948 other proteins 935:948 other proteins 935:948 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 5 57 theme cis-membrane 1017:1028 arg1 method 1043:1048 this cis-membrane FRET imaging method 1012:1048 this cis-membrane FRET imaging method 1012:1048 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 3 58 theme metabolic 540:548 arg1 labeling 557:564 metabolic glycan labeling 540:564 metabolic glycan labeling 540:564 We exploit metabolic glycan labeling in conjunction with site-specific protein labeling to simultaneously install a FRET acceptor and a donor onto the glycans and the extracellular terminal of the protein of interest, respectively. 24308457 1 59 theme powerful 214:221 arg1 tool 223:226 a powerful tool 212:226 a powerful tool for imaging glycome 212:246 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 1 59 theme powerful 214:221 arg1 labeling 97:104 Metabolic labeling 87:104 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry 87:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 4 60 theme intramolecular 765:778 arg1 distance 795:802 The intramolecular donor-acceptor distance 761:802 The intramolecular donor-acceptor distance for the POI 761:814 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 5 61 gly sialylation 1292:1302 arg1 integrin 1330:1337 β2 integrin activation 1327:1348 β2 integrin activation 1327:1348 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 62 theme several 1084:1090 arg1 receptors 1115:1123 several important cell surface receptors 1084:1123 several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1084:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 62 theme several 1084:1090 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 62 theme several 1084:1090 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 62 theme several 1084:1090 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 1 63 from labeling 97:104 arg1 conjunction 168:178 conjunction 168:178 conjunction with bioorthogonal chemistry 168:207 Metabolic labeling of glycans with chemical reproters (e.g., alkyne or azide) in conjunction with bioorthogonal chemistry is a powerful tool for imaging glycome; however, this method lacks protein-specificity and therefore is not applicable to imaging glycosylation of a specific protein of interest (POI). 24308457 2 64 from cells 522:526 arg1 imaging 495:501 protein-specific imaging 478:501 protein-specific imaging of glycans on live cells 478:526 Here we report the development of a cis-membrane FRET-based methodology that allows protein-specific imaging of glycans on live cells. 24308457 0 65 theme glycans 78:84 arg1 imaging 54:60 protein-specific imaging 37:60 protein-specific imaging of cell-surface glycans 37:84 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 24308457 6 66 theme cell-surface 1472:1483 arg1 proteins 1485:1492 various cell-surface proteins 1464:1492 various cell-surface proteins 1464:1492 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 5 67 gly sialylation 1069:1079 arg1 receptors 1115:1123 several important cell surface receptors 1084:1123 several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. 1084:1235 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 67 gly sialylation 1069:1079 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 67 gly sialylation 1069:1079 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 67 gly sialylation 1069:1079 arg1 factor-beta 1208:1218 transforming growth factor-beta 1188:1218 transforming growth factor-beta 1188:1218 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 4 68 from donor 971:975 arg1 distant 954:960 distant 954:960 distant 954:960 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 4 68 from donor 971:975 arg1 acceptors 922:930 the excess acceptors 911:930 the excess acceptors on other proteins 911:948 The intramolecular donor-acceptor distance for the POI falls within the range for effective FRET, whereas the intermolecular FRET is disfavored since the excess acceptors on other proteins are distant from the donor. 24308457 0 69 theme cell-surface 65:76 arg1 glycans 78:84 cell-surface glycans 65:84 cell-surface glycans 65:84 A cis-membrane FRET-based method for protein-specific imaging of cell-surface glycans. 24308457 5 70 theme integrin 1135:1142 arg1 αXβ2 1144:1147 integrin αXβ2 1135:1147 integrin αXβ2 1135:1147 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 6 71 theme various 1464:1470 arg1 proteins 1485:1492 various cell-surface proteins 1464:1492 various cell-surface proteins 1464:1492 Our methodology should enable the live-cell studies on how glycosylation regulates the functions and dynamics of various cell-surface proteins. 24308457 5 72 theme imaging 1254:1260 arg1 experiments 1262:1272 our imaging experiments 1250:1272 our imaging experiments 1250:1272 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 24308457 5 73 theme factor 1167:1172 arg1 receptor 1174:1181 epidermal growth factor receptor 1150:1181 epidermal growth factor receptor 1150:1181 We demonstrated the capability of this cis-membrane FRET imaging method by visualizing the sialylation of several important cell surface receptors including integrin αXβ2, epidermal growth factor receptor, and transforming growth factor-beta receptor type I. Furthermore, our imaging experiments revealed that the sialylation might be important for β2 integrin activation. 27601469 7 0 theme Å 1178:1178 arg1 resolution 1180:1189 1.80-2.32 Å resolution 1168:1189 1.80-2.32 Å resolution 1168:1189 Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. 27601469 3 1 theme glycobiomarker 691:704 arg1 detection 706:714 cancer glycobiomarker detection 684:714 cancer glycobiomarker detection 684:714 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 10 2 attach presence 1520:1527 arg2 forms 1546:1550 two isolectin forms 1532:1550 two isolectin forms 1532:1550 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 10 2 attach presence 1520:1527 arg1 WFA 1578:1580 commercially available WFA 1555:1580 commercially available WFA differing only in the identities of two amino acids 1555:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 5 3 theme WFA 875:877 arg1 structure 862:870 structure 862:870 structure 862:870 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 5 3 theme WFA 875:877 arg1 affinities 847:856 affinities 847:856 affinities 847:856 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 11 4 from macrocarpa 1735:1744 arg1 complex 1749:1755 complex 1749:1755 complex with GalNAc 1749:1767 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 10 5 theme isolectin 1536:1544 arg1 forms 1546:1550 two isolectin forms 1532:1550 two isolectin forms 1532:1550 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 3 6 theme leukemia 600:607 arg1 prostate 610:617 leukemia, prostate 600:617 prostate 610:617 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 1 7 theme specific 177:184 arg1 epitopes 199:206 specific carbohydrate epitopes 177:206 specific carbohydrate epitopes 177:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 7 8 with complex 1111:1117 arg1 GalNAc 1137:1142 GalNAc 1137:1142 GalNAc 1137:1142 Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. 27601469 7 8 with complex 1111:1117 arg1 LacdiNAc 1124:1131 LacdiNAc 1124:1131 LacdiNAc 1124:1131 Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. 27601469 8 9 theme resolution 1203:1212 arg1 structures 1214:1223 These high resolution structures 1192:1223 These high resolution structures 1192:1223 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 1 10 theme epitopes 199:206 arg1 hallmark 213:220 a hallmark 211:220 a hallmark of many cancers 211:236 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 10 theme epitopes 199:206 arg1 glycosylation 137:149 Aberrant glycosylation 128:149 Aberrant glycosylation 128:149 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 10 theme epitopes 199:206 arg1 overexpression 159:172 the overexpression 155:172 the overexpression of specific carbohydrate epitopes 155:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 10 11 theme available 1568:1576 arg1 WFA 1578:1580 commercially available WFA 1555:1580 commercially available WFA differing only in the identities of two amino acids 1555:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 0 12 theme floribunda 105:114 arg1 Agglutinin 116:125 Wisteria floribunda Agglutinin 96:125 Wisteria floribunda Agglutinin 96:125 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 3 13 theme pancreatic 620:629 arg1 cancers 651:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 11 14 attach isolated 1712:1719 arg1 macrocarpa 1735:1744 Vatairea macrocarpa 1726:1744 Vatairea macrocarpa 1726:1744 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 14 attach isolated 1712:1719 arg2 lectin 1705:1710 a homologous lectin 1692:1710 a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1692:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 9 15 theme surface 1405:1411 arg1 contribution 1366:1377 the contribution 1362:1377 the contribution of this small hydrophobic surface 1362:1411 Remarkably, the contribution of this small hydrophobic surface significantly increases the observed affinity for LacdiNAc over GalNAc. 27601469 8 16 theme hydrophobic 1236:1246 arg1 groove 1248:1253 a hydrophobic groove 1234:1253 a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring 1234:1347 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 3 17 theme prostate 610:617 arg1 cancers 651:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 8 18 theme complementary 1255:1267 arg1 groove 1248:1253 a hydrophobic groove 1234:1253 a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring 1234:1347 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 3 19 from malignancy 586:595 arg1 cancers 651:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 1 20 gly glycosylation 137:149 arg1 epitopes 199:206 specific carbohydrate epitopes 177:206 specific carbohydrate epitopes 177:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 9 21 theme observed 1441:1448 arg1 affinity 1450:1457 the observed affinity 1437:1457 the observed affinity for LacdiNAc 1437:1470 Remarkably, the contribution of this small hydrophobic surface significantly increases the observed affinity for LacdiNAc over GalNAc. 27601469 3 22 theme ovarian 632:638 arg1 cancers 651:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 1 23 theme Aberrant 128:135 arg1 hallmark 213:220 a hallmark 211:220 a hallmark of many cancers 211:236 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 23 theme Aberrant 128:135 arg1 glycosylation 137:149 Aberrant glycosylation 128:149 Aberrant glycosylation 128:149 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 23 theme Aberrant 128:135 arg1 overexpression 159:172 the overexpression 155:172 the overexpression of specific carbohydrate epitopes 155:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 11 24 with complex 1749:1755 arg1 GalNAc 1762:1767 GalNAc 1762:1767 GalNAc 1762:1767 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 6 25 theme surface 978:984 arg1 resonance 994:1002 surface plasmon resonance 978:1002 surface plasmon resonance 978:1002 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 8 26 theme minor 1293:1297 arg1 extent 1299:1304 a minor extent 1291:1304 a minor extent 1291:1304 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 6 27 theme × 1047:1047 arg1 m 1054:1054 9.24 × 10-5 m 1042:1054 9.24 × 10-5 m 1042:1054 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 0 28 theme Molecular 0:8 arg1 Basis 10:14 Molecular Basis 0:14 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 0:126 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 2 29 theme high 460:463 arg1 affinity 465:472 high affinity 460:472 high affinity 460:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 4 30 theme LacdiNAc 769:776 arg1 recognition 754:764 WFA recognition 750:764 WFA recognition of LacdiNAc 750:776 The mechanism of specificity for WFA recognition of LacdiNAc is not fully understood. 27601469 10 31 theme amino 1622:1626 arg1 acids 1628:1632 two amino acids 1618:1632 two amino acids 1618:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 11 32 theme Ser/Thr 1830:1836 arg1 αGalNAc 1847:1853 terminal Ser/Thr O-linked αGalNAc 1821:1853 not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1795:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 32 theme Ser/Thr 1830:1836 arg1 antigen 1859:1865 Tn antigen 1856:1865 Tn antigen 1856:1865 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 6 33 theme 4.67 1027:1030 arg1 m 1039:1039 4.67 × 10-4 m 1027:1039 4.67 × 10-4 m 1027:1039 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 8 34 theme GlcNAc 1331:1336 arg1 ring 1344:1347 the GlcNAc sugar ring 1327:1347 the GlcNAc sugar ring 1327:1347 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 2 35 theme floribunda 358:367 arg1 lectin 398:403 a legume lectin 389:403 a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity 389:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 2 35 theme floribunda 358:367 arg1 WFA 381:383 WFA 381:383 WFA 381:383 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 2 35 theme floribunda 358:367 arg1 agglutinin 369:378 Wisteria floribunda agglutinin 349:378 Wisteria floribunda agglutinin (WFA) 349:384 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 0 36 theme Cancer 39:44 arg1 LacdiNAc 62:69 LacdiNAc 62:69 LacdiNAc (GalNAc[β1→4]GlcNAc) 62:90 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 0 36 theme Cancer 39:44 arg1 Glycobiomarker 46:59 the Cancer Glycobiomarker 35:59 the Cancer Glycobiomarker 35:59 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 6 37 theme m 1073:1073 arg1 values 1017:1022 KD values 1014:1022 KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m 1014:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 11 38 theme binding 1665:1671 arg1 similar 1681:1687 similar 1681:1687 similar 1681:1687 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 38 theme binding 1665:1671 arg1 site 1673:1676 the WFA carbohydrate binding site 1644:1676 the WFA carbohydrate binding site 1644:1676 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 8 39 theme ring 1344:1347 arg1 back-face 1314:1322 the back-face 1310:1322 the back-face of the GlcNAc sugar ring 1310:1347 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 3 40 theme disaccharide 504:515 arg1 -d-GlcNAc 543:551 β-d-GalNAc-[1→4]-d-GlcNAc 527:551 β-d-GalNAc-[1→4]-d-GlcNAc 527:551 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 3 40 theme disaccharide 504:515 arg1 LacdiNAc 517:524 the disaccharide LacdiNAc 500:524 the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc) 500:552 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 10 41 from presence 1520:1527 arg1 WFA 1578:1580 commercially available WFA 1555:1580 commercially available WFA differing only in the identities of two amino acids 1555:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 6 42 theme × 1066:1066 arg1 m 1073:1073 5.45 × 10-6 m 1061:1073 5.45 × 10-6 m 1061:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 10 43 theme MS 1492:1493 arg1 sequencing 1495:1504 Tandem MS sequencing 1485:1504 Tandem MS sequencing 1485:1504 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 11 44 theme WFA 1648:1650 arg1 similar 1681:1687 similar 1681:1687 similar 1681:1687 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 44 theme WFA 1648:1650 arg1 site 1673:1676 the WFA carbohydrate binding site 1644:1676 the WFA carbohydrate binding site 1644:1676 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 1 45 theme many 225:228 arg1 cancers 230:236 many cancers 225:236 many cancers 225:236 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 11 46 link O-linked 1838:1845 arg1 αGalNAc 1847:1853 terminal Ser/Thr O-linked αGalNAc 1821:1853 not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1795:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 46 link O-linked 1838:1845 arg1 antigen 1859:1865 Tn antigen 1856:1865 Tn antigen 1856:1865 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 6 47 theme m 1054:1054 arg1 values 1017:1022 KD values 1014:1022 KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m 1014:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 1 48 theme tumor-associated 243:258 arg1 targets 306:312 targets 306:312 targets for immunotherapy and diagnostics 306:346 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 48 theme tumor-associated 243:258 arg1 oligosaccharides 260:275 tumor-associated oligosaccharides 243:275 tumor-associated oligosaccharides 243:275 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 2 49 with N-acetylgalactosaminides 430:453 arg1 affinity 465:472 high affinity 460:472 high affinity 460:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 10 50 theme forms 1546:1550 arg1 presence 1520:1527 the presence 1516:1527 the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids 1516:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 0 51 theme Wisteria 96:103 arg1 Agglutinin 116:125 Wisteria floribunda Agglutinin 96:125 Wisteria floribunda Agglutinin 96:125 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 3 52 theme tumor 580:584 arg1 malignancy 586:595 tumor malignancy 580:595 tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers 580:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 1 53 theme carbohydrate 186:197 arg1 epitopes 199:206 specific carbohydrate epitopes 177:206 specific carbohydrate epitopes 177:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 8 54 theme high 1198:1201 arg1 structures 1214:1223 These high resolution structures 1192:1223 These high resolution structures 1192:1223 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 11 55 theme Vatairea 1726:1733 arg1 macrocarpa 1735:1744 Vatairea macrocarpa 1726:1744 Vatairea macrocarpa 1726:1744 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 9 56 theme hydrophobic 1393:1403 arg1 surface 1405:1411 this small hydrophobic surface 1382:1411 this small hydrophobic surface 1382:1411 Remarkably, the contribution of this small hydrophobic surface significantly increases the observed affinity for LacdiNAc over GalNAc. 27601469 11 57 theme Tn 1856:1857 arg1 αGalNAc 1847:1853 terminal Ser/Thr O-linked αGalNAc 1821:1853 not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1795:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 57 theme Tn 1856:1857 arg1 antigen 1859:1865 Tn antigen 1856:1865 Tn antigen 1856:1865 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 5 58 from structure 862:870 arg1 complex 882:888 complex 882:888 complex with GalNAc and LacdiNAc 882:913 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 3 59 theme cancer 684:689 arg1 detection 706:714 cancer glycobiomarker detection 684:714 cancer glycobiomarker detection 684:714 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 5 60 from affinities 847:856 arg1 complex 882:888 complex 882:888 complex with GalNAc and LacdiNAc 882:913 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 4 61 theme specificity 734:744 arg1 mechanism 721:729 The mechanism 717:729 The mechanism of specificity for WFA recognition of LacdiNAc 717:776 The mechanism of specificity for WFA recognition of LacdiNAc is not fully understood. 27601469 6 62 theme KD 1014:1015 arg1 values 1017:1022 KD values 1014:1022 KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m 1014:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 7 63 from Structures 1090:1099 arg1 complex 1111:1117 complex 1111:1117 complex with LacdiNAc and GalNAc 1111:1142 Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. 27601469 6 64 theme plasmon 986:992 arg1 resonance 994:1002 surface plasmon resonance 978:1002 surface plasmon resonance 978:1002 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 11 65 theme homologous 1694:1703 arg1 lectin 1705:1710 a homologous lectin 1692:1710 a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1692:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 9 66 theme small 1387:1391 arg1 surface 1405:1411 this small hydrophobic surface 1382:1411 this small hydrophobic surface 1382:1411 Remarkably, the contribution of this small hydrophobic surface significantly increases the observed affinity for LacdiNAc over GalNAc. 27601469 11 67 theme terminal 1821:1828 arg1 αGalNAc 1847:1853 terminal Ser/Thr O-linked αGalNAc 1821:1853 not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1795:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 67 theme terminal 1821:1828 arg1 antigen 1859:1865 Tn antigen 1856:1865 Tn antigen 1856:1865 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 0 68 dep GalNAc[β1→4 72:82 arg1 GlcNAc 84:89 GlcNAc 84:89 GlcNAc 84:89 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 6 69 theme 9.24 1042:1045 arg1 m 1054:1054 9.24 × 10-5 m 1042:1054 9.24 × 10-5 m 1042:1054 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 4 70 theme WFA 750:752 arg1 recognition 754:764 WFA recognition 750:764 WFA recognition of LacdiNAc 750:776 The mechanism of specificity for WFA recognition of LacdiNAc is not fully understood. 27601469 7 71 theme WFA 1104:1106 arg1 Structures 1090:1099 Structures 1090:1099 Structures of WFA in complex with LacdiNAc and GalNAc 1090:1142 Structures of WFA in complex with LacdiNAc and GalNAc have been determined to 1.80-2.32 Å resolution. 27601469 6 72 theme m 1039:1039 arg1 values 1017:1022 KD values 1014:1022 KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m 1014:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 2 73 theme terminal 421:428 arg1 N-acetylgalactosaminides 430:453 terminal N-acetylgalactosaminides 421:453 terminal N-acetylgalactosaminides with high affinity 421:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 6 74 theme × 1032:1032 arg1 m 1039:1039 4.67 × 10-4 m 1027:1039 4.67 × 10-4 m 1027:1039 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 3 75 theme liver 645:649 arg1 cancers 651:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 leukemia, prostate, pancreatic, ovarian, and liver cancers 600:657 WFA preferentially binds the disaccharide LacdiNAc (β-d-GalNAc-[1→4]-d-GlcNAc), which is associated with tumor malignancy in leukemia, prostate, pancreatic, ovarian, and liver cancers and has shown promise in cancer glycobiomarker detection. 27601469 11 76 theme O-linked 1838:1845 arg1 αGalNAc 1847:1853 terminal Ser/Thr O-linked αGalNAc 1821:1853 not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen) 1795:1866 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 76 theme O-linked 1838:1845 arg1 antigen 1859:1865 Tn antigen 1856:1865 Tn antigen 1856:1865 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 0 77 theme Glycobiomarker 46:59 arg1 Recognition 20:30 Recognition 20:30 Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin 20:125 Molecular Basis for Recognition of the Cancer Glycobiomarker, LacdiNAc (GalNAc[β1→4]GlcNAc), by Wisteria floribunda Agglutinin. 27601469 5 78 with complex 882:888 arg1 LacdiNAc 906:913 LacdiNAc 906:913 LacdiNAc 906:913 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 5 78 with complex 882:888 arg1 GalNAc 895:900 GalNAc 895:900 GalNAc 895:900 To address this problem, we have determined affinities and structure of WFA in complex with GalNAc and LacdiNAc. 27601469 11 79 theme carbohydrate 1652:1663 arg1 similar 1681:1687 similar 1681:1687 similar 1681:1687 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 11 79 theme carbohydrate 1652:1663 arg1 site 1673:1676 the WFA carbohydrate binding site 1644:1676 the WFA carbohydrate binding site 1644:1676 Finally, the WFA carbohydrate binding site is similar to a homologous lectin isolated from Vatairea macrocarpa in complex with GalNAc, which, unlike WFA, binds not only αGalNAc but also terminal Ser/Thr O-linked αGalNAc (Tn antigen). 27601469 2 80 theme Wisteria 349:356 arg1 lectin 398:403 a legume lectin 389:403 a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity 389:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 2 80 theme Wisteria 349:356 arg1 WFA 381:383 WFA 381:383 WFA 381:383 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 2 80 theme Wisteria 349:356 arg1 agglutinin 369:378 Wisteria floribunda agglutinin 349:378 Wisteria floribunda agglutinin (WFA) 349:384 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 8 81 theme sugar 1338:1342 arg1 ring 1344:1347 the GlcNAc sugar ring 1327:1347 the GlcNAc sugar ring 1327:1347 These high resolution structures revealed a hydrophobic groove complementary to the GalNAc and, to a minor extent, to the back-face of the GlcNAc sugar ring. 27601469 10 82 theme acids 1628:1632 arg1 identities 1604:1613 the identities 1600:1613 the identities of two amino acids 1600:1632 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 6 83 theme 5.45 1061:1064 arg1 m 1073:1073 5.45 × 10-6 m 1061:1073 5.45 × 10-6 m 1061:1073 Affinities toward Gal, GalNAc, and LacdiNAc were measured via surface plasmon resonance, yielding KD values of 4.67 × 10-4 m, 9.24 × 10-5 m, and 5.45 × 10-6 m, respectively. 27601469 2 84 theme legume 391:396 arg1 lectin 398:403 a legume lectin 389:403 a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity 389:472 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 2 84 theme legume 391:396 arg1 agglutinin 369:378 Wisteria floribunda agglutinin 349:378 Wisteria floribunda agglutinin (WFA) 349:384 Wisteria floribunda agglutinin (WFA) is a legume lectin that recognizes terminal N-acetylgalactosaminides with high affinity. 27601469 10 85 theme Tandem 1485:1490 arg1 sequencing 1495:1504 Tandem MS sequencing 1485:1504 Tandem MS sequencing 1485:1504 Tandem MS sequencing confirmed the presence of two isolectin forms in commercially available WFA differing only in the identities of two amino acids. 27601469 1 86 theme cancers 230:236 arg1 hallmark 213:220 a hallmark 211:220 a hallmark of many cancers 211:236 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 86 theme cancers 230:236 arg1 glycosylation 137:149 Aberrant glycosylation 128:149 Aberrant glycosylation 128:149 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27601469 1 86 theme cancers 230:236 arg1 overexpression 159:172 the overexpression 155:172 the overexpression of specific carbohydrate epitopes 155:206 Aberrant glycosylation and the overexpression of specific carbohydrate epitopes is a hallmark of many cancers, and tumor-associated oligosaccharides are actively investigated as targets for immunotherapy and diagnostics. 27871459 9 0 from mesothelium 1262:1272 arg1 disc 1193:1196 the placental disc 1179:1196 the placental disc 1179:1196 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 7 1 gly glycosylated 989:1000 arg1 surfaces 1009:1016 glycosylated apical surfaces 989:1016 glycosylated apical surfaces expressing sialic acid and various other glycans 989:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 13 2 theme biochemical 1876:1886 arg1 conservation 1888:1899 biochemical conservation 1876:1899 biochemical conservation of this structure 1876:1917 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 12 3 theme near-term 1498:1506 arg1 placenta 1517:1524 near-term hedgehog placenta 1498:1524 near-term hedgehog placenta 1498:1524 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 11 4 theme endodermal 1453:1462 arg1 acini 1464:1468 endodermal acini 1453:1468 endodermal acini 1453:1468 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 7 5 contain had 985:987 arg1 endothelium 973:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 7 5 contain had 985:987 arg2 surfaces 1009:1016 glycosylated apical surfaces 989:1016 glycosylated apical surfaces expressing sialic acid and various other glycans 989:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 1 6 from findings 278:285 arg1 specimen 311:318 a near-term pregnant specimen 290:318 a near-term pregnant specimen 290:318 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 3 7 theme near 538:541 arg1 hoglets 548:554 four well-developed and near term hoglets 514:554 four well-developed and near term hoglets 514:554 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 12 8 theme DISCUSSION 1471:1480 arg1 features 1486:1493 DISCUSSION New features 1471:1493 DISCUSSION New features of near-term hedgehog placenta and associated tissues 1471:1547 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 11 9 theme inner 1357:1361 arg1 endoderm 1373:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 1 10 theme placenta 179:186 arg1 descriptions 159:170 few descriptions 155:170 few descriptions of the placenta 155:186 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 1 10 theme placenta 179:186 arg1 tissues 203:209 associated tissues 192:209 associated tissues of the European hedgehog (Erinaceus europaeus) 192:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 8 11 located present 1081:1087 arg2 Glycogen 1068:1075 Glycogen 1068:1075 Glycogen 1068:1075 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 8 11 located present 1081:1087 arg1 cells 1098:1102 large cells 1092:1102 large cells situated between the spongy zone and the endometrium 1092:1155 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 11 12 theme Yolk 1348:1351 arg1 sac 1353:1355 Yolk sac 1348:1355 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 0 13 theme Erinaceus 110:118 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 9 14 theme yolk 1240:1243 arg1 sac 1245:1247 yolk sac 1240:1247 yolk sac 1240:1247 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 5 15 theme glycan 849:854 arg1 classes 838:844 most classes 833:844 most classes of glycan 833:854 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 13 16 theme membrane 1793:1800 arg1 composition 1774:1784 the glycan composition 1763:1784 the glycan composition of the membrane itself 1763:1807 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 3 17 theme typical 604:610 arg1 labyrinth 612:620 typical labyrinth 604:620 typical labyrinth 604:620 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 7 18 theme various 1045:1051 arg1 glycans 1059:1065 various other glycans 1045:1065 various other glycans 1045:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 6 19 theme labyrinth 941:949 arg1 zone 951:954 the labyrinth zone 937:954 the labyrinth zone 937:954 Intercellular membranes were also glycosylated and this tended to be heavier in the labyrinth zone. 27871459 11 20 theme outer 1367:1371 arg1 endoderm 1373:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 8 21 from present 1081:1087 arg1 cells 1098:1102 large cells 1092:1102 large cells situated between the spongy zone and the endometrium 1092:1155 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 4 22 theme yolk 734:737 arg1 sac 739:741 a largely detached yolk sac 715:741 a largely detached yolk sac 715:741 In addition there was a paraplacenta incorporating Reichert's membrane and a largely detached yolk sac. 27871459 12 23 theme novel 1599:1603 arg1 sac 1610:1612 novel yolk sac 1599:1612 novel yolk sac acinar structures 1599:1630 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 7 24 theme apical 1002:1007 arg1 surfaces 1009:1016 glycosylated apical surfaces 989:1016 glycosylated apical surfaces expressing sialic acid and various other glycans 989:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 12 25 theme sac 1610:1612 arg1 structures 1621:1630 novel yolk sac acinar structures 1599:1630 novel yolk sac acinar structures 1599:1630 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 0 26 from histochemistry 12:25 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 27 theme tissues 76:82 arg1 histochemistry 12:25 histochemistry 12:25 histochemistry 12:25 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 27 theme tissues 76:82 arg1 Morphology 0:9 Morphology 0:9 Morphology 0:9 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 27 theme tissues 76:82 arg1 glycosylation 31:43 glycosylation 31:43 glycosylation 31:43 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 1 28 theme few 155:157 arg1 descriptions 159:170 few descriptions 155:170 few descriptions of the placenta 155:186 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 0 29 from Morphology 0:9 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 3 30 with placenta 590:597 arg1 labyrinth 612:620 typical labyrinth 604:620 typical labyrinth 604:620 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 3 30 with placenta 590:597 arg1 zones 633:637 spongy zones 626:637 spongy zones 626:637 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 7 31 theme capillary 963:971 arg1 endothelium 973:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 0 32 theme hedgehog 100:107 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 3 33 dep RESULTS 498:504 arg1 hoglets 548:554 four well-developed and near term hoglets 514:554 four well-developed and near term hoglets 514:554 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 3 33 dep RESULTS 498:504 arg1 Each 506:509 Each 506:509 Each 506:509 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 0 34 from glycosylation 31:43 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 8 35 theme large 1092:1096 arg1 cells 1098:1102 large cells 1092:1102 large cells situated between the spongy zone and the endometrium 1092:1155 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 1 36 dep INTRODUCTION 132:143 arg1 present 270:276 present 270:276 present findings on a near-term pregnant specimen 270:318 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 1 36 dep INTRODUCTION 132:143 arg1 are 151:153 are 151:153 are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) 151:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 6 37 gly glycosylated 891:902 arg1 membranes 871:879 Intercellular membranes 857:879 Intercellular membranes 857:879 Intercellular membranes were also glycosylated and this tended to be heavier in the labyrinth zone. 27871459 7 38 theme sialic 1029:1034 arg1 acid 1036:1039 sialic acid 1029:1039 sialic acid 1029:1039 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 8 39 theme situated 1104:1111 arg1 cells 1098:1102 large cells 1092:1102 large cells situated between the spongy zone and the endometrium 1092:1155 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 13 40 theme structure 1909:1917 arg1 conservation 1888:1899 biochemical conservation 1876:1899 biochemical conservation of this structure 1876:1917 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 13 41 theme placental 1670:1678 arg1 disc 1680:1683 the placental disc 1666:1683 the placental disc 1666:1683 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 9 42 theme Trophoblast 1158:1168 arg1 cells 1170:1174 Trophoblast cells 1158:1174 Trophoblast cells in the placental disc and under Reichert's membrane 1158:1226 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 5 43 theme granule 809:815 arg1 populations 794:804 diverse populations 786:804 diverse populations of granule which expressed most classes of glycan 786:854 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 1 44 theme near-term 292:300 arg1 specimen 311:318 a near-term pregnant specimen 290:318 a near-term pregnant specimen 290:318 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 6 45 theme Intercellular 857:869 arg1 membranes 871:879 Intercellular membranes 857:879 Intercellular membranes 857:879 Intercellular membranes were also glycosylated and this tended to be heavier in the labyrinth zone. 27871459 12 46 theme tissues 1541:1547 arg1 features 1486:1493 DISCUSSION New features 1471:1493 DISCUSSION New features of near-term hedgehog placenta and associated tissues 1471:1547 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 6 47 from heavier 926:932 arg1 zone 951:954 the labyrinth zone 937:954 the labyrinth zone 937:954 Intercellular membranes were also glycosylated and this tended to be heavier in the labyrinth zone. 27871459 9 48 theme placental 1183:1191 arg1 disc 1193:1196 the placental disc 1179:1196 the placental disc 1179:1196 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 1 49 theme hedgehog 227:234 arg1 descriptions 159:170 few descriptions 155:170 few descriptions of the placenta 155:186 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 1 49 theme hedgehog 227:234 arg1 tissues 203:209 associated tissues 192:209 associated tissues of the European hedgehog (Erinaceus europaeus) 192:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 9 50 dep sac 1245:1247 arg1 endoderm 1249:1256 endoderm 1249:1256 endoderm 1249:1256 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 11 51 theme similar 1392:1398 arg1 glycans 1400:1406 similar glycans 1392:1406 similar glycans 1392:1406 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 1 52 theme Erinaceus 237:245 arg1 hedgehog 227:234 the European hedgehog 214:234 the European hedgehog (Erinaceus europaeus) 214:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 1 52 theme Erinaceus 237:245 arg1 europaeus 247:255 Erinaceus europaeus 237:255 Erinaceus europaeus 237:255 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 12 53 theme hedgehog 1508:1515 arg1 placenta 1517:1524 near-term hedgehog placenta 1498:1524 near-term hedgehog placenta 1498:1524 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 3 54 theme term 543:546 arg1 hoglets 548:554 four well-developed and near term hoglets 514:554 four well-developed and near term hoglets 514:554 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 11 55 theme N-acetylgalactosamine 1419:1439 arg1 residues 1441:1448 N-acetylgalactosamine residues 1419:1448 N-acetylgalactosamine residues in endodermal acini 1419:1468 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 9 56 from sac 1245:1247 arg1 disc 1193:1196 the placental disc 1179:1196 the placental disc 1179:1196 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 12 57 theme New 1482:1484 arg1 features 1486:1493 DISCUSSION New features 1471:1493 DISCUSSION New features of near-term hedgehog placenta and associated tissues 1471:1547 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 11 58 theme sac 1353:1355 arg1 endoderm 1373:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm 1348:1380 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 3 59 theme discoid 568:574 arg1 placenta 590:597 a discoid, haemochorial placenta 566:597 a discoid, haemochorial placenta with typical labyrinth and spongy zones 566:637 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 10 60 gly glycosylated 1334:1345 arg1 membrane 1313:1320 Reichert's membrane 1302:1320 Reichert's membrane 1302:1320 Reichert's membrane was heavily glycosylated. 27871459 11 61 from residues 1441:1448 arg1 acini 1464:1468 endodermal acini 1453:1468 endodermal acini 1453:1468 Yolk sac inner and outer endoderm expressed similar glycans except for N-acetylgalactosamine residues in endodermal acini. 27871459 8 62 from cells 1098:1102 arg1 present 1081:1087 present 1081:1087 present 1081:1087 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 7 63 theme other 1053:1057 arg1 glycans 1059:1065 various other glycans 1045:1065 various other glycans 1045:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 3 64 theme spongy 626:631 arg1 zones 633:637 spongy zones 626:637 spongy zones 626:637 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 12 65 theme yolk 1605:1608 arg1 sac 1610:1612 novel yolk sac 1599:1612 novel yolk sac acinar structures 1599:1630 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 4 66 theme detached 725:732 arg1 sac 739:741 a largely detached yolk sac 715:741 a largely detached yolk sac 715:741 In addition there was a paraplacenta incorporating Reichert's membrane and a largely detached yolk sac. 27871459 0 67 theme associated 65:74 arg1 tissues 76:82 associated tissues 65:82 associated tissues 65:82 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 9 68 from cells 1170:1174 arg1 disc 1193:1196 the placental disc 1179:1196 the placental disc 1179:1196 Trophoblast cells in the placental disc and under Reichert's membrane, as well as yolk sac endoderm and mesothelium, were cytokeratin positive. 27871459 12 69 theme acinar 1614:1619 arg1 structures 1621:1630 novel yolk sac acinar structures 1599:1630 novel yolk sac acinar structures 1599:1630 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 5 70 contain contained 776:784 arg1 trophoblast 748:758 The trophoblast 744:758 The trophoblast of the placenta 744:774 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 5 70 contain contained 776:784 arg2 populations 794:804 diverse populations 786:804 diverse populations of granule which expressed most classes of glycan 786:854 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 5 71 theme placenta 767:774 arg1 trophoblast 748:758 The trophoblast 744:758 The trophoblast of the placenta 744:774 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 3 72 dep discoid 568:574 arg1 haemochorial 577:588 haemochorial 577:588 haemochorial 577:588 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 27871459 2 73 theme lectin 475:480 arg1 histochemistry 482:495 lectin histochemistry 475:495 lectin histochemistry 475:495 METHODS Tissues were examined grossly and then formalin fixed and wax-embedded for histology and immunocytochemistry (cytokeratin) and resin embedded for lectin histochemistry. 27871459 13 74 theme glycan 1767:1772 arg1 composition 1774:1784 the glycan composition 1763:1784 the glycan composition of the membrane itself 1763:1807 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 0 75 theme European 91:98 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 7 76 theme glycosylated 989:1000 arg1 surfaces 1009:1016 glycosylated apical surfaces 989:1016 glycosylated apical surfaces expressing sialic acid and various other glycans 989:1065 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 1 77 theme associated 192:201 arg1 tissues 203:209 associated tissues 192:209 associated tissues of the European hedgehog (Erinaceus europaeus) 192:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 13 78 from that 1721:1724 arg1 differences 1704:1714 significant differences 1692:1714 significant differences from that underlying Reichert's membrane 1692:1755 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 7 79 theme Fetal 957:961 arg1 endothelium 973:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium 957:983 Fetal capillary endothelium had glycosylated apical surfaces expressing sialic acid and various other glycans. 27871459 13 80 theme conservation 1888:1899 arg1 degree 1866:1871 a degree 1864:1871 a degree of biochemical conservation of this structure 1864:1917 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 5 81 theme most 833:836 arg1 classes 838:844 most classes 833:844 most classes of glycan 833:854 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 8 82 attach present 1081:1087 arg1 cells 1098:1102 large cells 1092:1102 large cells situated between the spongy zone and the endometrium 1092:1155 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 8 82 attach present 1081:1087 arg2 Glycogen 1068:1075 Glycogen 1068:1075 Glycogen 1068:1075 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 5 83 theme diverse 786:792 arg1 populations 794:804 diverse populations 786:804 diverse populations of granule which expressed most classes of glycan 786:854 The trophoblast of the placenta contained diverse populations of granule which expressed most classes of glycan. 27871459 8 84 theme spongy 1125:1130 arg1 zone 1132:1135 the spongy zone 1121:1135 the spongy zone 1121:1135 Glycogen was present in large cells situated between the spongy zone and the endometrium. 27871459 0 85 dep placenta 52:59 arg1 the 48:50 the 48:50 the 48:50 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 86 theme placenta 52:59 arg1 histochemistry 12:25 histochemistry 12:25 histochemistry 12:25 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 86 theme placenta 52:59 arg1 Morphology 0:9 Morphology 0:9 Morphology 0:9 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 86 theme placenta 52:59 arg1 glycosylation 31:43 glycosylation 31:43 glycosylation 31:43 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 13 87 theme disc 1680:1683 arg1 trophoblast 1651:1661 The trophoblast 1647:1661 The trophoblast of the placental disc 1647:1683 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 2 88 theme METHODS 321:327 arg1 Tissues 329:335 METHODS Tissues 321:335 METHODS Tissues 321:335 METHODS Tissues were examined grossly and then formalin fixed and wax-embedded for histology and immunocytochemistry (cytokeratin) and resin embedded for lectin histochemistry. 27871459 1 89 theme pregnant 302:309 arg1 specimen 311:318 a near-term pregnant specimen 290:318 a near-term pregnant specimen 290:318 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 12 90 theme placenta 1517:1524 arg1 features 1486:1493 DISCUSSION New features 1471:1493 DISCUSSION New features of near-term hedgehog placenta and associated tissues 1471:1547 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 0 91 gly glycosylation 31:43 arg1 tissues 76:82 associated tissues 65:82 associated tissues 65:82 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 91 gly glycosylation 31:43 arg1 europaeus 120:128 the European hedgehog (Erinaceus europaeus) 87:129 the European hedgehog (Erinaceus europaeus) 87:129 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 0 91 gly glycosylation 31:43 arg1 placenta 52:59 placenta 52:59 placenta 52:59 Morphology, histochemistry and glycosylation of the placenta and associated tissues in the European hedgehog (Erinaceus europaeus). 27871459 13 92 theme significant 1692:1702 arg1 differences 1704:1714 significant differences 1692:1714 significant differences from that underlying Reichert's membrane 1692:1755 The trophoblast of the placental disc showed significant differences from that underlying Reichert's membrane while the glycan composition of the membrane itself showed some similarity to that of rat thereby implying a degree of biochemical conservation of this structure. 27871459 1 93 theme European 218:225 arg1 hedgehog 227:234 the European hedgehog 214:234 the European hedgehog (Erinaceus europaeus) 214:256 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 1 93 theme European 218:225 arg1 europaeus 247:255 Erinaceus europaeus 237:255 Erinaceus europaeus 237:255 INTRODUCTION There are few descriptions of the placenta and associated tissues of the European hedgehog (Erinaceus europaeus) and here we present findings on a near-term pregnant specimen. 27871459 12 94 theme associated 1530:1539 arg1 tissues 1541:1547 associated tissues 1530:1547 associated tissues 1530:1547 DISCUSSION New features of near-term hedgehog placenta and associated tissues are presented, including their glycosylation, and novel yolk sac acinar structures are described. 27871459 2 95 dep fixed 377:381 arg1 grossly 351:357 grossly 351:357 grossly 351:357 METHODS Tissues were examined grossly and then formalin fixed and wax-embedded for histology and immunocytochemistry (cytokeratin) and resin embedded for lectin histochemistry. 27871459 3 96 theme well-developed 519:532 arg1 hoglets 548:554 four well-developed and near term hoglets 514:554 four well-developed and near term hoglets 514:554 RESULTS Each of four well-developed and near term hoglets displayed a discoid, haemochorial placenta with typical labyrinth and spongy zones. 26082228 0 0 theme O-Glycosylation 75:89 arg1 Modification 46:57 Modification 46:57 Modification of Plant N- and O-Glycosylation 46:89 Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation. 26082228 4 1 theme enzymes 544:550 arg1 retention 514:522 retention 514:522 retention 514:522 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 4 1 theme enzymes 544:550 arg1 targeting 500:508 targeting 500:508 targeting 500:508 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 1 2 theme recombinant 153:163 arg1 therapeutics 178:189 recombinant glycoprotein therapeutics 153:189 recombinant glycoprotein therapeutics 153:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 4 3 theme early 559:563 arg1 pathway 575:581 the early secretory pathway 555:581 the early secretory pathway of plants 555:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 5 4 theme glycosidases 692:703 arg1 localization 651:662 the subcellular localization 635:662 the subcellular localization of glycosyltransferases and glycosidases in plants 635:713 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 1 5 theme glycoprotein 165:176 arg1 therapeutics 178:189 recombinant glycoprotein therapeutics 153:189 recombinant glycoprotein therapeutics 153:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 5 6 theme subcellular 639:649 arg1 localization 651:662 the subcellular localization 635:662 the subcellular localization of glycosyltransferases and glycosidases in plants 635:713 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 3 7 theme customized 400:409 arg1 structures 442:451 customized and homogenous N- and O-glycan structures 400:451 customized and homogenous N- and O-glycan structures 400:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 3 8 theme recombinant 369:379 arg1 glycoproteins 381:393 recombinant glycoproteins 369:393 recombinant glycoproteins 369:393 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 5 9 theme glycosyltransferases 667:686 arg1 localization 651:662 the subcellular localization 635:662 the subcellular localization of glycosyltransferases and glycosidases in plants 635:713 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 1 10 theme therapeutics 178:189 arg1 production 139:148 the production 135:148 the production of recombinant glycoprotein therapeutics 135:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 3 11 theme glycoproteins 381:393 arg1 production 355:364 the production 351:364 the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures 351:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 4 12 theme glycan-modifying 527:542 arg1 enzymes 544:550 glycan-modifying enzymes 527:550 glycan-modifying enzymes 527:550 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 5 13 used used 617:620 arg2 protocols 600:608 These protocols 594:608 These protocols 594:608 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 3 14 theme N- 426:427 arg1 structures 442:451 customized and homogenous N- and O-glycan structures 400:451 customized and homogenous N- and O-glycan structures 400:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 4 15 from retention 514:522 arg1 pathway 575:581 the early secretory pathway 555:581 the early secretory pathway of plants 555:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 5 16 theme recombinant 780:790 arg1 glycoproteins 792:804 recombinant glycoproteins 780:804 recombinant glycoproteins 780:804 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 0 17 theme Subcellular 0:10 arg1 Targeting 12:20 Subcellular Targeting 0:20 Subcellular Targeting of Proteins 0:32 Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation. 26082228 0 18 theme Proteins 25:32 arg1 Targeting 12:20 Subcellular Targeting 0:20 Subcellular Targeting of Proteins 0:32 Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation. 26082228 4 19 from targeting 500:508 arg1 pathway 575:581 the early secretory pathway 555:581 the early secretory pathway of plants 555:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 3 20 theme homogenous 415:424 arg1 structures 442:451 customized and homogenous N- and O-glycan structures 400:451 customized and homogenous N- and O-glycan structures 400:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 1 21 theme attractive 103:112 arg1 Plants 92:97 Plants 92:97 Plants 92:97 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 1 21 theme attractive 103:112 arg1 hosts 125:129 attractive expression hosts 103:129 attractive expression hosts for the production of recombinant glycoprotein therapeutics 103:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 3 22 gly glycoproteins 381:393 arg1 glycoproteins 381:393 recombinant glycoproteins 369:393 recombinant glycoproteins 369:393 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 3 23 theme O-glycan 433:440 arg1 structures 442:451 customized and homogenous N- and O-glycan structures 400:451 customized and homogenous N- and O-glycan structures 400:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 4 24 theme secretory 565:573 arg1 pathway 575:581 the early secretory pathway 555:581 the early secretory pathway of plants 555:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 1 25 theme expression 114:123 arg1 Plants 92:97 Plants 92:97 Plants 92:97 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 1 25 theme expression 114:123 arg1 hosts 125:129 attractive expression hosts 103:129 attractive expression hosts for the production of recombinant glycoprotein therapeutics 103:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 5 26 gly homogeneity 748:758 arg1 glycosylation 763:775 glycosylation 763:775 glycosylation 763:775 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 4 27 theme plants 586:591 arg1 pathway 575:581 the early secretory pathway 555:581 the early secretory pathway of plants 555:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 5 28 theme glycosylation 763:775 arg1 homogeneity 748:758 the homogeneity 744:758 the homogeneity of glycosylation on recombinant glycoproteins 744:804 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 2 29 theme biopharmaceuticals 228:245 arg1 efficiency 208:217 efficiency 208:217 efficiency 208:217 The quality and efficiency of these biopharmaceuticals are very often influenced by the glycosylation profile. 26082228 2 29 theme biopharmaceuticals 228:245 arg1 quality 196:202 quality 196:202 quality 196:202 The quality and efficiency of these biopharmaceuticals are very often influenced by the glycosylation profile. 26082228 1 30 gly glycoprotein 165:176 arg1 glycoprotein 165:176 recombinant glycoprotein therapeutics 153:189 recombinant glycoprotein therapeutics 153:189 Plants are attractive expression hosts for the production of recombinant glycoprotein therapeutics. 26082228 2 31 theme glycosylation 280:292 arg1 profile 294:300 the glycosylation profile 276:300 the glycosylation profile 276:300 The quality and efficiency of these biopharmaceuticals are very often influenced by the glycosylation profile. 26082228 5 32 gly glycoproteins 792:804 arg1 glycoproteins 792:804 recombinant glycoproteins 780:804 recombinant glycoproteins 780:804 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 5 33 from homogeneity 748:758 arg1 glycoproteins 792:804 recombinant glycoproteins 780:804 recombinant glycoproteins 780:804 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 4 34 theme convenient 472:481 arg1 tools 483:487 convenient tools 472:487 convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants 472:591 Here, we describe convenient tools that allow targeting and retention of glycan-modifying enzymes in the early secretory pathway of plants. 26082228 0 35 theme N- 68:69 arg1 Modification 46:57 Modification 46:57 Modification of Plant N- and O-Glycosylation 46:89 Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation. 26082228 3 36 with production 355:364 arg1 structures 442:451 customized and homogenous N- and O-glycan structures 400:451 customized and homogenous N- and O-glycan structures 400:451 Consequently, approaches are needed that enable the production of recombinant glycoproteins with customized and homogenous N- and O-glycan structures. 26082228 5 37 from localization 651:662 arg1 plants 708:713 plants 708:713 plants 708:713 These protocols can be used to fine-tune the subcellular localization of glycosyltransferases and glycosidases in plants and consequently to increase the homogeneity of glycosylation on recombinant glycoproteins. 26082228 0 38 theme Plant 62:66 arg1 N- 68:69 Plant N- 62:69 Plant N- 62:69 Subcellular Targeting of Proteins Involved in Modification of Plant N- and O-Glycosylation. 26582281 5 0 theme core 824:827 arg1 N-glycan 829:836 this core N-glycan 819:836 this core N-glycan 819:836 Additional decorations occur on this core N-glycan and represent a molecular signature for each chlorovirus. 26582281 2 1 with pentasaccharide 352:366 arg1 β-glucose 375:383 a β-glucose 373:383 a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S 373:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 2 theme common 318:323 arg1 structure 307:315 This N-glycan core structure 288:315 This N-glycan core structure 288:315 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 2 theme common 318:323 arg1 pentasaccharide 352:366 a pentasaccharide 350:366 a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S 350:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 3 located located 430:436 arg2 located 430:436 located 430:436 located 430:436 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 3 located located 430:436 arg2 residue 409:415 an asparagine residue 395:415 an asparagine residue which is not located in the typical sequon N-X-T/S 395:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 3 located located 430:436 arg1 N-X-T/S 460:466 the typical sequon N-X-T/S 441:466 the typical sequon N-X-T/S 441:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 3 4 theme second 606:611 arg1 residue 620:626 a second xylose residue 604:626 a second xylose residue 604:626 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 1 5 theme new 241:243 arg1 type 245:248 a new type 239:248 a new type of core N-glycan 239:265 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 3 6 theme xylose 613:618 arg1 residue 620:626 a second xylose residue 604:626 a second xylose residue 604:626 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 5 7 theme molecular 854:862 arg1 signature 864:872 a molecular signature 852:872 a molecular signature for each chlorovirus 852:893 Additional decorations occur on this core N-glycan and represent a molecular signature for each chlorovirus. 26582281 2 8 from located 430:436 arg1 N-X-T/S 460:466 the typical sequon N-X-T/S 441:466 the typical sequon N-X-T/S 441:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 4 9 theme absolute 744:751 arg1 configuration 753:765 its absolute configuration 740:765 its absolute configuration 740:765 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 2 10 theme asparagine 398:407 arg1 residue 409:415 an asparagine residue 395:415 an asparagine residue which is not located in the typical sequon N-X-T/S 395:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 10 theme asparagine 398:407 arg1 located 430:436 located 430:436 located 430:436 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 1 11 theme life 174:177 arg1 life 174:177 life 174:177 life 174:177 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 1 11 theme life 174:177 arg1 domains 163:169 the three domains 153:169 the three domains of life 153:177 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 4 12 theme unit 662:665 arg1 position 639:646 The third position 629:646 The third position of the fucose unit 629:665 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 0 13 theme N-Linked 0:7 arg1 Glycans 9:15 N-Linked Glycans 0:15 N-Linked Glycans of Chloroviruses 0:32 N-Linked Glycans of Chloroviruses Sharing a Core Architecture without Precedent. 26582281 4 14 attach linked 677:682 arg1 element 724:730 a semiconserved element 708:730 a semiconserved element 708:730 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 4 14 attach linked 677:682 arg2 position 639:646 The third position 629:646 The third position of the fucose unit 629:665 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 4 14 attach linked 677:682 arg1 rhamnose 689:696 a rhamnose 687:696 a rhamnose 687:696 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 4 15 theme fucose 655:660 arg1 unit 662:665 the fucose unit 651:665 the fucose unit 651:665 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 3 16 theme terminal 581:588 arg1 galactose 590:598 a terminal galactose 579:598 a terminal galactose 579:598 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 0 17 theme Chloroviruses 20:32 arg1 Glycans 9:15 N-Linked Glycans 0:15 N-Linked Glycans of Chloroviruses 0:32 N-Linked Glycans of Chloroviruses Sharing a Core Architecture without Precedent. 26582281 1 18 theme core 253:256 arg1 N-glycan 258:265 core N-glycan 253:265 core N-glycan 253:265 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 1 19 theme fundamental 102:112 arg1 N-glycosylation 81:95 N-glycosylation 81:95 N-glycosylation 81:95 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 1 19 theme fundamental 102:112 arg1 modification 114:125 a fundamental modification 100:125 a fundamental modification of proteins 100:137 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 1 20 theme N-glycan 258:265 arg1 type 245:248 a new type 239:248 a new type of core N-glycan 239:265 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 3 21 theme xylose 505:510 arg1 unit 512:515 a terminal xylose unit 494:515 a terminal xylose unit 494:515 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 3 22 attach linked 484:489 arg2 glucose 473:479 The glucose 469:479 The glucose 469:479 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 3 22 attach linked 484:489 arg1 fucose 537:542 a hyperbranched fucose 521:542 a hyperbranched fucose 521:542 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 3 22 attach linked 484:489 arg1 unit 512:515 a terminal xylose unit 494:515 a terminal xylose unit 494:515 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 4 23 theme third 633:637 arg1 position 639:646 The third position 629:646 The third position of the fucose unit 629:665 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 2 24 attach linked 385:390 arg1 residue 409:415 an asparagine residue 395:415 an asparagine residue which is not located in the typical sequon N-X-T/S 395:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 24 attach linked 385:390 arg2 β-glucose 375:383 a β-glucose 373:383 a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S 373:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 24 attach linked 385:390 arg1 located 430:436 located 430:436 located 430:436 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 4 25 theme semiconserved 710:722 arg1 element 724:730 a semiconserved element 708:730 a semiconserved element 708:730 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 4 25 theme semiconserved 710:722 arg1 rhamnose 689:696 a rhamnose 687:696 a rhamnose 687:696 The third position of the fucose unit is always linked to a rhamnose, which is a semiconserved element because its absolute configuration is virus-dependent. 26582281 0 26 theme Core 44:47 arg1 Architecture 49:60 a Core Architecture 42:60 a Core Architecture 42:60 N-Linked Glycans of Chloroviruses Sharing a Core Architecture without Precedent. 26582281 3 27 theme hyperbranched 523:535 arg1 fucose 537:542 a hyperbranched fucose 521:542 a hyperbranched fucose 521:542 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 1 28 theme proteins 130:137 arg1 N-glycosylation 81:95 N-glycosylation 81:95 N-glycosylation 81:95 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 1 28 theme proteins 130:137 arg1 modification 114:125 a fundamental modification 100:125 a fundamental modification of proteins 100:137 N-glycosylation is a fundamental modification of proteins and exists in the three domains of life and in some viruses, including the chloroviruses, for which a new type of core N-glycan is herein described. 26582281 2 29 from N-X-T/S 460:466 arg1 residue 409:415 an asparagine residue 395:415 an asparagine residue which is not located in the typical sequon N-X-T/S 395:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 29 from N-X-T/S 460:466 arg1 located 430:436 located 430:436 located 430:436 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 30 theme core 302:305 arg1 structure 307:315 This N-glycan core structure 288:315 This N-glycan core structure 288:315 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 30 theme core 302:305 arg1 pentasaccharide 352:366 a pentasaccharide 350:366 a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S 350:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 3 31 theme terminal 496:503 arg1 unit 512:515 a terminal xylose unit 494:515 a terminal xylose unit 494:515 The glucose is linked to a terminal xylose unit and a hyperbranched fucose, which is in turn substituted with a terminal galactose and a second xylose residue. 26582281 2 32 theme N-glycan 293:300 arg1 structure 307:315 This N-glycan core structure 288:315 This N-glycan core structure 288:315 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 32 theme N-glycan 293:300 arg1 pentasaccharide 352:366 a pentasaccharide 350:366 a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S 350:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 2 33 theme sequon 453:458 arg1 N-X-T/S 460:466 the typical sequon N-X-T/S 441:466 the typical sequon N-X-T/S 441:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 26582281 5 34 theme Additional 787:796 arg1 decorations 798:808 Additional decorations 787:808 Additional decorations 787:808 Additional decorations occur on this core N-glycan and represent a molecular signature for each chlorovirus. 26582281 2 35 theme typical 445:451 arg1 N-X-T/S 460:466 the typical sequon N-X-T/S 441:466 the typical sequon N-X-T/S 441:466 This N-glycan core structure, common to all chloroviruses, is a pentasaccharide with a β-glucose linked to an asparagine residue which is not located in the typical sequon N-X-T/S. 27500750 5 0 dep % 784:784 arg1 capacity 835:842 strong anti-interference capacity 810:842 strong anti-interference capacity 810:842 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 5 0 dep % 784:784 arg1 biosamples 795:804 real biosamples 790:804 real biosamples 790:804 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 4 1 theme hydropathy-index-based 547:568 arg1 strategy 570:577 a hydropathy-index-based strategy 545:577 a hydropathy-index-based strategy for sequence optimization 545:603 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 5 2 dep selectivity 763:773 arg1 e.g. 757:760 e.g. 757:760 e.g. 757:760 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 5 3 theme 1000-fold 865:873 arg1 albumin 888:894 1000-fold bovine serum albumin 865:894 1000-fold bovine serum albumin interference 865:907 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 4 4 theme sequence 583:590 arg1 optimization 592:603 sequence optimization 583:603 sequence optimization 583:603 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 2 5 theme mass 409:412 arg1 spectrometry 414:425 mass spectrometry 409:425 mass spectrometry 409:425 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 8 6 theme future 1423:1428 arg1 development 1430:1440 future development 1423:1440 future development of glycoproteomics and glycobiology 1423:1476 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 6 7 theme different 1042:1050 arg1 compositions 1052:1063 different compositions 1042:1063 different compositions 1042:1063 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 5 8 dep ∼70 781:783 arg1 to 778:779 to 778:779 to 778:779 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 4 9 from types 662:666 arg1 sequences 644:652 three optimal dipeptide sequences 620:652 three optimal dipeptide sequences from 54 types of dipeptides 620:680 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 2 10 theme complex 335:341 arg1 biosamples 343:352 complex biosamples 335:352 complex biosamples 335:352 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 2 11 gly glycopeptides 316:328 arg2 glycopeptides 316:328 low-abundance glycopeptides 302:328 low-abundance glycopeptides from complex biosamples 302:352 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 2 11 gly glycopeptides 316:328 arg1 biosamples 343:352 complex biosamples 335:352 complex biosamples 335:352 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 5 12 theme dipeptide-based 697:711 arg1 homopolymers 713:724 The optimized dipeptide-based homopolymers 683:724 The optimized dipeptide-based homopolymers 683:724 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 1 13 from impact 186:191 arg1 biofunctions 222:233 biofunctions 222:233 biofunctions 222:233 Glycoproteomics identifies and catalogs protein glycosylation and explores its impact on protein conformations and biofunctions. 27500750 1 13 from impact 186:191 arg1 conformations 204:216 protein conformations 196:216 protein conformations 196:216 Glycoproteomics identifies and catalogs protein glycosylation and explores its impact on protein conformations and biofunctions. 27500750 5 14 dep display 726:732 arg1 selectivity 763:773 selectivity 763:773 selectivity 763:773 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 3 15 theme binding 493:499 arg1 platform 501:508 a simple but promising carbohydrate binding platform 457:508 a simple but promising carbohydrate binding platform to tackle these problems 457:533 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 3 15 theme binding 493:499 arg1 dipeptide 444:452 dipeptide 444:452 dipeptide 444:452 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 5 16 theme bovine 875:880 arg1 albumin 888:894 1000-fold bovine serum albumin 865:894 1000-fold bovine serum albumin interference 865:907 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 8 17 theme glycan 1355:1360 arg1 structures 1362:1371 the complex glycan structures 1343:1371 the complex glycan structures 1343:1371 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 5 18 theme anti-interference 817:833 arg1 capacity 835:842 strong anti-interference capacity 810:842 strong anti-interference capacity 810:842 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 0 19 theme Carbohydrate 16:27 arg1 Receptors 29:37 Dipeptide-Based Carbohydrate Receptors 0:37 Dipeptide-Based Carbohydrate Receptors 0:37 Dipeptide-Based Carbohydrate Receptors and Polymers for Glycopeptide Enrichment and Glycan Discrimination. 27500750 6 20 theme linkage 1104:1110 arg1 isomers 1112:1118 their linkage isomers 1098:1118 their linkage isomers 1098:1118 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 5 21 theme capable 844:850 arg1 biosamples 795:804 real biosamples 790:804 real biosamples 790:804 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 3 22 theme promising 470:478 arg1 platform 501:508 a simple but promising carbohydrate binding platform 457:508 a simple but promising carbohydrate binding platform to tackle these problems 457:533 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 3 22 theme promising 470:478 arg1 dipeptide 444:452 dipeptide 444:452 dipeptide 444:452 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 0 23 theme Dipeptide-Based 0:14 arg1 Receptors 29:37 Dipeptide-Based Carbohydrate Receptors 0:37 Dipeptide-Based Carbohydrate Receptors 0:37 Dipeptide-Based Carbohydrate Receptors and Polymers for Glycopeptide Enrichment and Glycan Discrimination. 27500750 4 24 theme optimal 626:632 arg1 sequences 644:652 three optimal dipeptide sequences 620:652 three optimal dipeptide sequences from 54 types of dipeptides 620:680 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 5 25 theme excellent 734:742 arg1 performance 744:754 excellent performance 734:754 excellent performance 734:754 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 7 26 theme isomeric 1260:1267 arg1 structures 1219:1228 subtly variable glycan structures 1196:1228 subtly variable glycan structures of glycopeptides 1196:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 7 26 theme isomeric 1260:1267 arg1 linkages 1280:1287 isomeric glycosidic linkages 1260:1287 isomeric glycosidic linkages 1260:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 3 27 theme carbohydrate 480:491 arg1 platform 501:508 a simple but promising carbohydrate binding platform 457:508 a simple but promising carbohydrate binding platform to tackle these problems 457:533 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 3 27 theme carbohydrate 480:491 arg1 dipeptide 444:452 dipeptide 444:452 dipeptide 444:452 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 5 28 theme glycopeptide 913:924 arg1 enrichment 926:935 glycopeptide enrichment 913:935 glycopeptide enrichment 913:935 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 7 29 theme glycopeptides 1233:1245 arg1 linkages 1280:1287 isomeric glycosidic linkages 1260:1287 isomeric glycosidic linkages 1260:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 7 29 theme glycopeptides 1233:1245 arg1 structures 1219:1228 subtly variable glycan structures 1196:1228 subtly variable glycan structures of glycopeptides 1196:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 5 30 theme optimized 687:695 arg1 homopolymers 713:724 The optimized dipeptide-based homopolymers 683:724 The optimized dipeptide-based homopolymers 683:724 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 7 31 theme glycosidic 1269:1278 arg1 structures 1219:1228 subtly variable glycan structures 1196:1228 subtly variable glycan structures of glycopeptides 1196:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 7 31 theme glycosidic 1269:1278 arg1 linkages 1280:1287 isomeric glycosidic linkages 1260:1287 isomeric glycosidic linkages 1260:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 6 32 theme chromatographic 986:1000 arg1 separation 1002:1011 high-efficiency chromatographic separation 970:1011 high-efficiency chromatographic separation 970:1011 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 1 33 theme protein 196:202 arg1 conformations 204:216 protein conformations 196:216 protein conformations 196:216 Glycoproteomics identifies and catalogs protein glycosylation and explores its impact on protein conformations and biofunctions. 27500750 8 34 theme glycoproteomics 1445:1459 arg1 development 1430:1440 future development 1423:1440 future development of glycoproteomics and glycobiology 1423:1476 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 3 35 theme simple 459:464 arg1 platform 501:508 a simple but promising carbohydrate binding platform 457:508 a simple but promising carbohydrate binding platform to tackle these problems 457:533 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 3 35 theme simple 459:464 arg1 dipeptide 444:452 dipeptide 444:452 dipeptide 444:452 Here, we report dipeptide as a simple but promising carbohydrate binding platform to tackle these problems. 27500750 4 36 theme dipeptide 634:642 arg1 sequences 644:652 three optimal dipeptide sequences 620:652 three optimal dipeptide sequences from 54 types of dipeptides 620:680 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 5 37 theme real 790:793 arg1 biosamples 795:804 real biosamples 790:804 real biosamples 790:804 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 6 38 theme high-efficiency 970:984 arg1 separation 1002:1011 high-efficiency chromatographic separation 970:1011 high-efficiency chromatographic separation 970:1011 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 6 39 with oligosaccharides 1020:1035 arg1 isomers 1112:1118 their linkage isomers 1098:1118 their linkage isomers 1098:1118 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 6 39 with oligosaccharides 1020:1035 arg1 degrees 1081:1087 polymerization degrees 1066:1087 polymerization degrees 1066:1087 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 6 39 with oligosaccharides 1020:1035 arg1 compositions 1052:1063 different compositions 1042:1063 different compositions 1042:1063 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 5 40 theme serum 882:886 arg1 albumin 888:894 1000-fold bovine serum albumin 865:894 1000-fold bovine serum albumin interference 865:907 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 8 41 theme solid 1315:1319 arg1 foundation 1321:1330 a solid foundation 1313:1330 a solid foundation to analyze the complex glycan structures and glycosites simultaneously 1313:1401 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 8 42 theme glycobiology 1465:1476 arg1 development 1430:1440 future development 1423:1440 future development of glycoproteomics and glycobiology 1423:1476 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 5 43 theme albumin 888:894 arg1 interference 896:907 1000-fold bovine serum albumin interference 865:907 1000-fold bovine serum albumin interference 865:907 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 7 44 theme variable 1203:1210 arg1 linkages 1280:1287 isomeric glycosidic linkages 1260:1287 isomeric glycosidic linkages 1260:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 7 44 theme variable 1203:1210 arg1 structures 1219:1228 subtly variable glycan structures 1196:1228 subtly variable glycan structures of glycopeptides 1196:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 1 45 theme protein 147:153 arg1 glycosylation 155:167 protein glycosylation 147:167 protein glycosylation 147:167 Glycoproteomics identifies and catalogs protein glycosylation and explores its impact on protein conformations and biofunctions. 27500750 0 46 theme Glycopeptide 56:67 arg1 Enrichment 69:78 Glycopeptide Enrichment 56:78 Glycopeptide Enrichment 56:78 Dipeptide-Based Carbohydrate Receptors and Polymers for Glycopeptide Enrichment and Glycan Discrimination. 27500750 7 47 theme glycan 1212:1217 arg1 linkages 1280:1287 isomeric glycosidic linkages 1260:1287 isomeric glycosidic linkages 1260:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 7 47 theme glycan 1212:1217 arg1 structures 1219:1228 subtly variable glycan structures 1196:1228 subtly variable glycan structures of glycopeptides 1196:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 5 48 gly glycopeptide 913:924 arg2 glycopeptide 913:924 glycopeptide enrichment 913:935 glycopeptide enrichment 913:935 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27500750 8 49 gly glycosites 1377:1386 arg2 glycosites 1377:1386 glycosites 1377:1386 glycosites 1377:1386 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 2 50 theme low-abundance 302:314 arg1 glycopeptides 316:328 low-abundance glycopeptides 302:328 low-abundance glycopeptides from complex biosamples 302:352 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 2 51 theme glycan 388:393 arg1 structures 395:404 glycan structures 388:404 glycan structures 388:404 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 7 52 theme attractive 1141:1150 arg1 feature 1152:1158 another attractive feature 1133:1158 another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages 1133:1287 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 8 53 dep provide 1305:1311 arg1 benefit 1415:1421 benefit 1415:1421 will benefit future development of glycoproteomics and glycobiology 1410:1476 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 2 54 from biosamples 343:352 arg1 glycopeptides 316:328 low-abundance glycopeptides 302:328 low-abundance glycopeptides from complex biosamples 302:352 However, these studies are restricted by the bottleneck to enrich low-abundance glycopeptides from complex biosamples and the difficulties in analyzing glycan structures by mass spectrometry. 27500750 4 55 theme dipeptides 671:680 arg1 types 662:666 54 types 659:666 54 types of dipeptides 659:680 We build a hydropathy-index-based strategy for sequence optimization and screen out three optimal dipeptide sequences from 54 types of dipeptides. 27500750 0 56 theme Glycan 84:89 arg1 Discrimination 91:104 Glycan Discrimination 84:104 Glycan Discrimination 84:104 Dipeptide-Based Carbohydrate Receptors and Polymers for Glycopeptide Enrichment and Glycan Discrimination. 27500750 8 57 theme complex 1347:1353 arg1 structures 1362:1371 the complex glycan structures 1343:1371 the complex glycan structures 1343:1371 These features provide a solid foundation to analyze the complex glycan structures and glycosites simultaneously, which will benefit future development of glycoproteomics and glycobiology. 27500750 6 58 theme polymerization 1066:1079 arg1 degrees 1081:1087 polymerization degrees 1066:1087 polymerization degrees 1066:1087 Meanwhile, our polymers exhibit high-efficiency chromatographic separation toward oligosaccharides with different compositions, polymerization degrees and even their linkage isomers. 27500750 7 59 gly glycopeptides 1233:1245 arg2 glycopeptides 1233:1245 glycopeptides 1233:1245 glycopeptides 1233:1245 This brings another attractive feature that our materials can discriminate subtly variable glycan structures of glycopeptides, especially, isomeric glycosidic linkages. 27500750 5 60 theme strong 810:815 arg1 capacity 835:842 strong anti-interference capacity 810:842 strong anti-interference capacity 810:842 The optimized dipeptide-based homopolymers display excellent performance (e.g., selectivity up to ∼70% for real biosamples and strong anti-interference capacity capable of resisting 1000-fold bovine serum albumin interference) in glycopeptide enrichment. 27283789 1 0 gly glycosylated 96:107 arg1 proteins 109:116 homogeneously glycosylated proteins 82:116 homogeneously glycosylated proteins 82:116 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 27283789 5 1 theme glycodendrimer 740:753 arg1 structures 755:764 novel glycodendrimer structures 734:764 novel glycodendrimer structures 734:764 These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. 27283789 4 2 theme growth 616:621 arg1 hormone 623:629 human growth hormone 610:629 human growth hormone 610:629 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 4 3 theme human 610:614 arg1 hormone 623:629 human growth hormone 610:629 human growth hormone 610:629 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 5 4 theme new 690:692 arg1 class 694:698 a new class 688:698 a new class of engineered biosimilars bearing novel glycodendrimer structures 688:764 These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. 27283789 2 5 theme glycoprotein 347:358 arg1 mimics 360:365 'N-glycosylated' glycoprotein mimics 330:365 'N-glycosylated' glycoprotein mimics 330:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 2 6 gly glycoprotein 347:358 arg1 glycoprotein 347:358 'N-glycosylated' glycoprotein mimics 330:365 'N-glycosylated' glycoprotein mimics 330:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 4 7 theme chemoenzymatic 573:586 arg1 route 588:592 a chemoenzymatic route 571:592 a chemoenzymatic route 571:592 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 5 8 theme engineered 703:712 arg1 biosimilars 714:724 engineered biosimilars 703:724 engineered biosimilars bearing novel glycodendrimer structures 703:764 These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. 27283789 2 9 theme -glycosylated 332:344 arg1 mimics 360:365 'N-glycosylated' glycoprotein mimics 330:365 'N-glycosylated' glycoprotein mimics 330:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 2 10 gly -glycosylated 332:344 arg1 mimics 360:365 'N-glycosylated' glycoprotein mimics 330:365 'N-glycosylated' glycoprotein mimics 330:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 5 11 theme biosimilars 714:724 arg1 class 694:698 a new class 688:698 a new class of engineered biosimilars bearing novel glycodendrimer structures 688:764 These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. 27283789 1 12 theme glycosylated 96:107 arg1 proteins 109:116 homogeneously glycosylated proteins 82:116 homogeneously glycosylated proteins 82:116 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 27283789 1 13 theme proteins 109:116 arg1 generation 68:77 The generation 64:77 The generation of homogeneously glycosylated proteins 64:116 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 27283789 1 13 theme proteins 109:116 arg1 essential 121:129 essential 121:129 essential 121:129 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 27283789 5 14 theme novel 734:738 arg1 structures 755:764 novel glycodendrimer structures 734:764 novel glycodendrimer structures 734:764 These products represent a new class of engineered biosimilars bearing novel glycodendrimer structures. 27283789 3 15 theme protein 437:443 arg1 point 456:460 protein attachment point 437:460 protein attachment point 437:460 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 3 16 theme attachment 445:454 arg1 point 456:460 protein attachment point 437:460 protein attachment point 437:460 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 4 17 theme IgG 658:660 arg1 hormone 623:629 human growth hormone 610:629 human growth hormone 610:629 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 4 17 theme IgG 658:660 arg1 region 642:647 the Fc region 635:647 the Fc region of human IgG 635:660 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 2 18 theme novel 227:231 arg1 prosthetic 246:255 a novel glycodendron prosthetic 225:255 a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics 225:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 3 19 theme native 478:483 arg1 architecture 494:505 the native N-glycan architecture 474:505 the native N-glycan architecture 474:505 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 4 20 theme human 652:656 arg1 IgG 658:660 human IgG 652:660 human IgG 652:660 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 0 21 theme N-glycan 42:49 arg1 prosthetics 51:61 protein N-glycan prosthetics 34:61 protein N-glycan prosthetics 34:61 Piperidine-based glycodendrons as protein N-glycan prosthetics. 27283789 3 22 theme N-glycan 485:492 arg1 architecture 494:505 the native N-glycan architecture 474:505 the native N-glycan architecture 474:505 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 0 23 theme protein 34:40 arg1 prosthetics 51:61 protein N-glycan prosthetics 34:61 protein N-glycan prosthetics 34:61 Piperidine-based glycodendrons as protein N-glycan prosthetics. 27283789 3 24 theme dendrimer 414:422 arg1 scaffold 424:431 the dendrimer scaffold 410:431 the dendrimer scaffold 410:431 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 4 25 attach attached 598:605 arg1 hormone 623:629 human growth hormone 610:629 human growth hormone 610:629 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 4 25 attach attached 598:605 arg2 glycodendrimers 534:548 Three piperidine-melamine glycodendrimers 508:548 Three piperidine-melamine glycodendrimers 508:548 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 4 25 attach attached 598:605 arg1 region 642:647 the Fc region 635:647 the Fc region of human IgG 635:660 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 4 26 theme Fc 639:640 arg1 region 642:647 the Fc region 635:647 the Fc region of human IgG 635:660 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 3 27 theme computational 374:386 arg1 modeling 388:395 computational modeling 374:395 computational modeling 374:395 Using computational modeling, we designed the dendrimer scaffold and protein attachment point to resemble the native N-glycan architecture. 27283789 1 28 theme glycoform-specific 144:161 arg1 activity 163:170 glycoform-specific activity 144:170 glycoform-specific activity 144:170 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 27283789 2 29 theme 'N 330:331 arg1 mimics 360:365 'N-glycosylated' glycoprotein mimics 330:365 'N-glycosylated' glycoprotein mimics 330:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 4 30 theme piperidine-melamine 514:532 arg1 glycodendrimers 534:548 Three piperidine-melamine glycodendrimers 508:548 Three piperidine-melamine glycodendrimers 508:548 Three piperidine-melamine glycodendrimers were synthesized via a chemoenzymatic route and attached to human growth hormone and the Fc region of human IgG. 27283789 2 31 theme glycodendron 233:244 arg1 prosthetic 246:255 a novel glycodendron prosthetic 225:255 a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics 225:365 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 2 32 theme recombinant 299:309 arg1 proteins 311:318 recombinant proteins 299:318 recombinant proteins 299:318 We present a novel glycodendron prosthetic which can be site-selectively appended to recombinant proteins to create 'N-glycosylated' glycoprotein mimics. 27283789 1 33 theme protein-based 186:198 arg1 therapeutics 200:211 protein-based therapeutics 186:211 protein-based therapeutics 186:211 The generation of homogeneously glycosylated proteins is essential for defining glycoform-specific activity and improving protein-based therapeutics. 29157724 8 0 theme useful 1143:1148 arg1 type 1109:1112 high mannose type and sialylated N-glycans 1096:1137 type 1109:1112 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 0 theme useful 1143:1148 arg1 N-glycans 1129:1137 high mannose type and sialylated N-glycans 1096:1137 N-glycans 1129:1137 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 0 theme useful 1143:1148 arg1 discriminators 1150:1163 useful discriminators 1143:1163 useful discriminators 1143:1163 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 1 dep characteristic 1049:1062 arg1 ROC 1065:1067 ROC 1065:1067 ROC 1065:1067 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 7 2 theme high 862:865 arg1 N-glycans 880:888 high mannose type N-glycans 862:888 high mannose type N-glycans 862:888 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 9 3 theme cancer 1434:1439 arg1 tissues 1441:1447 cancer tissues 1434:1447 cancer tissues 1434:1447 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 12 4 theme lung 2081:2084 arg1 adenocarcinoma 2086:2099 lung adenocarcinoma 2081:2099 lung adenocarcinoma 2081:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 11 5 theme tissue 1725:1730 arg1 sections 1732:1739 formalin-fixed paraffin-embedded (FFPE) tissue sections 1685:1739 formalin-fixed paraffin-embedded (FFPE) tissue sections 1685:1739 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 2 6 theme fresh 307:311 arg1 samples 320:326 fresh tissue samples 307:326 fresh tissue samples 307:326 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 12 7 theme differential 2047:2058 arg1 patterns 2069:2076 differential N-glycan patterns 2047:2076 differential N-glycan patterns of lung adenocarcinoma 2047:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 10 8 theme several 1571:1577 arg1 processes 1598:1606 several physiopathological processes 1571:1606 several physiopathological processes 1571:1606 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 10 8 theme several 1571:1577 arg1 carcinogenesis 1619:1632 carcinogenesis 1619:1632 carcinogenesis 1619:1632 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 8 9 theme control 1212:1218 arg1 tissue 1220:1225 control tissue 1212:1225 control tissue 1212:1225 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 0 10 theme tissue 130:135 arg1 sections 137:144 formalin-fixed paraffin-embedded (FFPE) tissue sections 90:144 formalin-fixed paraffin-embedded (FFPE) tissue sections 90:144 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 4 11 dep paraffin-embedded 579:595 arg1 FFPE 598:601 FFPE 598:601 FFPE 598:601 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 4 12 theme adenocarcinoma 630:643 arg1 patients 645:652 lung adenocarcinoma patients 625:652 lung adenocarcinoma patients 625:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 7 13 theme control 1009:1015 arg1 samples 1017:1023 the control samples 1005:1023 the control samples 1005:1023 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 6 14 used used 808:811 arg2 compositions 731:742 58 N-glycan compositions 719:742 58 N-glycan compositions 719:742 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 6 14 used used 808:811 arg2 51 783:784 51 783:784 51 783:784 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 9 15 theme expression 1264:1273 arg1 levels 1275:1280 expression levels 1264:1280 expression levels of specific N-glycans 1264:1302 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 11 16 theme expressed 1816:1824 arg1 N-glycans 1826:1834 the differentially expressed N-glycans 1797:1834 the differentially expressed N-glycans of lung adenocarcinoma 1797:1857 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 1 17 theme N-glycan 147:154 arg1 approach 180:187 a powerful approach 169:187 a powerful approach for analyzing the functional relationship between N-glycosylation and cancer 169:264 N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. 29157724 1 17 theme N-glycan 147:154 arg1 profiling 156:164 N-glycan profiling 147:164 N-glycan profiling 147:164 N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. 29157724 0 18 theme N-glycan 68:75 arg1 profiling 77:85 N-glycan profiling 68:85 N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections 68:144 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 9 19 theme N-glycans 1294:1302 arg1 levels 1275:1280 expression levels 1264:1280 expression levels of specific N-glycans 1264:1302 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 7 20 theme adenocarcinoma 969:982 arg1 samples 984:990 our FFPE lung adenocarcinoma samples 955:990 our FFPE lung adenocarcinoma samples 955:990 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 11 21 theme adenocarcinoma 1844:1857 arg1 N-glycans 1826:1834 the differentially expressed N-glycans 1797:1834 the differentially expressed N-glycans of lung adenocarcinoma 1797:1857 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 12 22 theme clinical 1936:1943 arg1 diagnosis 1945:1953 clinical diagnosis 1936:1953 clinical diagnosis as well as identification of potential biomarkers 1936:2003 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 12 23 theme biomarkers 1994:2003 arg1 diagnosis 1945:1953 clinical diagnosis 1936:1953 clinical diagnosis as well as identification of potential biomarkers 1936:2003 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 12 23 theme biomarkers 1994:2003 arg1 identification 1966:1979 identification 1966:1979 clinical diagnosis as well as identification of potential biomarkers 1936:2003 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 7 24 theme FFPE 959:962 arg1 samples 984:990 our FFPE lung adenocarcinoma samples 955:990 our FFPE lung adenocarcinoma samples 955:990 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 1 25 theme powerful 171:178 arg1 approach 180:187 a powerful approach 169:187 a powerful approach for analyzing the functional relationship between N-glycosylation and cancer 169:264 N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. 29157724 1 25 theme powerful 171:178 arg1 profiling 156:164 N-glycan profiling 147:164 N-glycan profiling 147:164 N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. 29157724 7 26 dep downregulated 938:950 arg1 compared 993:1000 compared 993:1000 compared to the control samples 993:1023 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 9 27 theme lung 1324:1327 arg1 adenocarcinoma 1329:1342 lung adenocarcinoma 1324:1342 lung adenocarcinoma 1324:1342 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 8 28 theme receiver 1030:1037 arg1 analysis 1076:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 11 29 from changes 1753:1759 arg1 patterns 1773:1780 N-glycan patterns 1764:1780 N-glycan patterns 1764:1780 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 10 30 theme important 1496:1504 arg1 modifications 1533:1545 the most important post-translational protein modifications 1487:1545 the most important post-translational protein modifications 1487:1545 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 8 31 theme characteristic 1049:1062 arg1 analysis 1076:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 0 32 dep paraffin-embedded 105:121 arg1 FFPE 124:127 FFPE 124:127 FFPE 124:127 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 2 33 theme tissues 446:452 arg1 tissues 446:452 tissues 446:452 tissues 446:452 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 2 33 theme tissues 446:452 arg1 variety 435:441 a variety 433:441 a variety of tissues 433:452 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 0 34 theme Differential 0:11 arg1 patterns 22:29 Differential N-glycan patterns 0:29 Differential N-glycan patterns 0:29 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 7 35 theme sialylated 913:922 arg1 N-glycans 924:932 sialylated N-glycans 913:932 sialylated N-glycans 913:932 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 11 36 dep paraffin-embedded 1700:1716 arg1 FFPE 1719:1722 FFPE 1719:1722 FFPE 1719:1722 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 8 37 theme curve 1070:1074 arg1 analysis 1076:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 6 38 from samples 774:780 arg1 compositions 731:742 58 N-glycan compositions 719:742 58 N-glycan compositions 719:742 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 4 39 theme profiling 538:546 arg1 method 548:553 a profiling method 536:553 a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients 536:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 10 40 theme protein 1525:1531 arg1 modifications 1533:1545 the most important post-translational protein modifications 1487:1545 the most important post-translational protein modifications 1487:1545 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 2 41 theme serum 412:416 arg1 proteins 400:407 the proteins 396:407 the proteins of serum 396:416 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 9 42 theme FFPE-based 1375:1384 arg1 method 1386:1391 our FFPE-based method 1371:1391 our FFPE-based method 1371:1391 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 12 43 theme FFPE-based 1885:1894 arg1 useful 1925:1930 useful 1925:1930 useful 1925:1930 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 12 43 theme FFPE-based 1885:1894 arg1 method 1915:1920 the FFPE-based N-glycan profiling method 1881:1920 the FFPE-based N-glycan profiling method 1881:1920 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 6 44 theme adenocarcinoma 754:767 arg1 samples 774:780 lung adenocarcinoma FFPE samples 749:780 lung adenocarcinoma FFPE samples 749:780 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 8 45 theme high 1096:1099 arg1 type 1109:1112 high mannose type and sialylated N-glycans 1096:1137 type 1109:1112 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 45 theme high 1096:1099 arg1 N-glycans 1129:1137 high mannose type and sialylated N-glycans 1096:1137 N-glycans 1129:1137 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 45 theme high 1096:1099 arg1 discriminators 1150:1163 useful discriminators 1143:1163 useful discriminators 1143:1163 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 12 46 theme profiling 1905:1913 arg1 useful 1925:1930 useful 1925:1930 useful 1925:1930 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 12 46 theme profiling 1905:1913 arg1 method 1915:1920 the FFPE-based N-glycan profiling method 1881:1920 the FFPE-based N-glycan profiling method 1881:1920 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 3 47 theme tissue 474:479 arg1 samples 481:487 fresh tissue samples 468:487 fresh tissue samples 468:487 Furthermore, fresh tissue samples are difficult to ship and store. 29157724 6 48 theme MSn-based 817:825 arg1 prediction 837:846 MSn-based structure prediction 817:846 MSn-based structure prediction 817:846 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 6 49 theme FFPE 769:772 arg1 samples 774:780 lung adenocarcinoma FFPE samples 749:780 lung adenocarcinoma FFPE samples 749:780 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 6 50 theme N-glycan 722:729 arg1 compositions 731:742 58 N-glycan compositions 719:742 58 N-glycan compositions 719:742 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 9 51 theme N-glycan 1412:1419 arg1 profiling 1421:1429 N-glycan profiling 1412:1429 N-glycan profiling of cancer tissues 1412:1447 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 8 52 theme sialylated 1118:1127 arg1 type 1109:1112 high mannose type and sialylated N-glycans 1096:1137 type 1109:1112 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 52 theme sialylated 1118:1127 arg1 N-glycans 1129:1137 high mannose type and sialylated N-glycans 1096:1137 N-glycans 1129:1137 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 52 theme sialylated 1118:1127 arg1 discriminators 1150:1163 useful discriminators 1143:1163 useful discriminators 1143:1163 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 7 53 gly sialylated 913:922 arg1 N-glycans 924:932 sialylated N-glycans 913:932 sialylated N-glycans 913:932 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 10 54 theme physiopathological 1579:1596 arg1 processes 1598:1606 several physiopathological processes 1571:1606 several physiopathological processes 1571:1606 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 10 54 theme physiopathological 1579:1596 arg1 carcinogenesis 1619:1632 carcinogenesis 1619:1632 carcinogenesis 1619:1632 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 11 55 theme formalin-fixed 1685:1698 arg1 sections 1732:1739 formalin-fixed paraffin-embedded (FFPE) tissue sections 1685:1739 formalin-fixed paraffin-embedded (FFPE) tissue sections 1685:1739 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 4 56 used used 531:534 arg2 we 528:529 we 528:529 we 528:529 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 2 57 theme tissue 313:318 arg1 samples 320:326 fresh tissue samples 307:326 fresh tissue samples 307:326 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 7 58 theme mannose 867:873 arg1 N-glycans 880:888 high mannose type N-glycans 862:888 high mannose type N-glycans 862:888 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 9 59 theme tissues 1441:1447 arg1 profiling 1421:1429 N-glycan profiling 1412:1429 N-glycan profiling of cancer tissues 1412:1447 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 4 60 theme formalin-fixed 564:577 arg1 sections 611:618 formalin-fixed paraffin-embedded (FFPE) tissue sections 564:618 formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients 564:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 12 61 theme adenocarcinoma 2086:2099 arg1 patterns 2069:2076 differential N-glycan patterns 2047:2076 differential N-glycan patterns of lung adenocarcinoma 2047:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 4 62 from patients 645:652 arg1 sections 611:618 formalin-fixed paraffin-embedded (FFPE) tissue sections 564:618 formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients 564:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 8 63 theme lung 1188:1191 arg1 adenocarcinoma 1193:1206 lung adenocarcinoma 1188:1206 lung adenocarcinoma 1188:1206 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 0 64 theme sections 137:144 arg1 profiling 77:85 N-glycan profiling 68:85 N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections 68:144 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 1 65 theme functional 207:216 arg1 relationship 218:229 the functional relationship 203:229 the functional relationship between N-glycosylation and cancer 203:264 N-glycan profiling is a powerful approach for analyzing the functional relationship between N-glycosylation and cancer. 29157724 12 66 theme N-glycan 2060:2067 arg1 patterns 2069:2076 differential N-glycan patterns 2047:2076 differential N-glycan patterns of lung adenocarcinoma 2047:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 4 67 dep formalin-fixed 564:577 arg1 paraffin-embedded 579:595 paraffin-embedded 579:595 paraffin-embedded 579:595 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 2 68 theme Current 267:273 arg1 methods 275:281 Current methods 267:281 Current methods 267:281 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 12 69 theme patterns 2069:2076 arg1 knowledge 2034:2042 current knowledge 2026:2042 current knowledge of differential N-glycan patterns of lung adenocarcinoma 2026:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 4 70 theme lung 625:628 arg1 patients 645:652 lung adenocarcinoma patients 625:652 lung adenocarcinoma patients 625:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 12 71 theme current 2026:2032 arg1 knowledge 2034:2042 current knowledge 2026:2042 current knowledge of differential N-glycan patterns of lung adenocarcinoma 2026:2099 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 9 72 theme specific 1285:1292 arg1 N-glycans 1294:1302 specific N-glycans 1285:1302 specific N-glycans 1285:1302 Together, our results indicate that expression levels of specific N-glycans correlate well with lung adenocarcinoma, and strongly suggest that our FFPE-based method will be useful for N-glycan profiling of cancer tissues. 29157724 0 73 theme formalin-fixed 90:103 arg1 sections 137:144 formalin-fixed paraffin-embedded (FFPE) tissue sections 90:144 formalin-fixed paraffin-embedded (FFPE) tissue sections 90:144 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 8 74 theme operating 1039:1047 arg1 analysis 1076:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis 1026:1083 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 7 75 theme lung 964:967 arg1 samples 984:990 our FFPE lung adenocarcinoma samples 955:990 our FFPE lung adenocarcinoma samples 955:990 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 29157724 10 76 theme SIGNIFICANCE 1450:1461 arg1 Glycosylation 1463:1475 SIGNIFICANCE Glycosylation 1450:1475 SIGNIFICANCE Glycosylation 1450:1475 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 11 77 theme lung 1839:1842 arg1 adenocarcinoma 1844:1857 lung adenocarcinoma 1839:1857 lung adenocarcinoma 1839:1857 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 0 78 theme N-glycan 13:20 arg1 patterns 22:29 Differential N-glycan patterns 0:29 Differential N-glycan patterns 0:29 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 12 79 theme potential 1984:1992 arg1 biomarkers 1994:2003 potential biomarkers 1984:2003 potential biomarkers 1984:2003 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 0 80 dep formalin-fixed 90:103 arg1 paraffin-embedded 105:121 paraffin-embedded 105:121 paraffin-embedded 105:121 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 11 81 dep formalin-fixed 1685:1698 arg1 paraffin-embedded 1700:1716 paraffin-embedded 1700:1716 paraffin-embedded 1700:1716 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 10 82 theme modifications 1533:1545 arg1 modifications 1533:1545 the most important post-translational protein modifications 1487:1545 the most important post-translational protein modifications 1487:1545 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 10 82 theme modifications 1533:1545 arg1 one 1480:1482 one 1480:1482 one 1480:1482 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 3 83 theme fresh 468:472 arg1 samples 481:487 fresh tissue samples 468:487 fresh tissue samples 468:487 Furthermore, fresh tissue samples are difficult to ship and store. 29157724 0 84 theme lung 45:48 arg1 adenocarcinoma 50:63 lung adenocarcinoma 45:63 lung adenocarcinoma 45:63 Differential N-glycan patterns identified in lung adenocarcinoma by N-glycan profiling of formalin-fixed paraffin-embedded (FFPE) tissue sections. 29157724 10 85 theme post-translational 1506:1523 arg1 modifications 1533:1545 the most important post-translational protein modifications 1487:1545 the most important post-translational protein modifications 1487:1545 SIGNIFICANCE Glycosylation is one of the most important post-translational protein modifications, and is associated with several physiopathological processes, including carcinogenesis. 29157724 8 86 gly sialylated 1118:1127 arg1 type 1109:1112 high mannose type and sialylated N-glycans 1096:1137 type 1109:1112 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 86 gly sialylated 1118:1127 arg1 N-glycans 1129:1137 high mannose type and sialylated N-glycans 1096:1137 N-glycans 1129:1137 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 86 gly sialylated 1118:1127 arg1 discriminators 1150:1163 useful discriminators 1143:1163 useful discriminators 1143:1163 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 87 theme mannose 1101:1107 arg1 type 1109:1112 high mannose type and sialylated N-glycans 1096:1137 type 1109:1112 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 87 theme mannose 1101:1107 arg1 N-glycans 1129:1137 high mannose type and sialylated N-glycans 1096:1137 N-glycans 1129:1137 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 8 87 theme mannose 1101:1107 arg1 discriminators 1150:1163 useful discriminators 1143:1163 useful discriminators 1143:1163 Our receiver operating characteristic (ROC) curve analysis shows that high mannose type and sialylated N-glycans are useful discriminators to distinguish between lung adenocarcinoma and control tissue. 29157724 6 88 theme lung 749:752 arg1 samples 774:780 lung adenocarcinoma FFPE samples 749:780 lung adenocarcinoma FFPE samples 749:780 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 6 89 theme structure 827:835 arg1 prediction 837:846 MSn-based structure prediction 817:846 MSn-based structure prediction 817:846 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 12 90 theme N-glycan 1896:1903 arg1 useful 1925:1930 useful 1925:1930 useful 1925:1930 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 12 90 theme N-glycan 1896:1903 arg1 method 1915:1920 the FFPE-based N-glycan profiling method 1881:1920 the FFPE-based N-glycan profiling method 1881:1920 Our study shows that the FFPE-based N-glycan profiling method is useful for clinical diagnosis as well as identification of potential biomarkers, and our data expand current knowledge of differential N-glycan patterns of lung adenocarcinoma. 29157724 11 91 theme N-glycan 1764:1771 arg1 patterns 1773:1780 N-glycan patterns 1764:1780 N-glycan patterns 1764:1780 In this study, we tested the feasibility of using formalin-fixed paraffin-embedded (FFPE) tissue sections to identify changes in N-glycan patterns and identified the differentially expressed N-glycans of lung adenocarcinoma. 29157724 6 92 theme compositions 731:742 arg1 51 783:784 51 783:784 51 783:784 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 6 92 theme compositions 731:742 arg1 compositions 731:742 58 N-glycan compositions 719:742 58 N-glycan compositions 719:742 We identified 58 N-glycan compositions from lung adenocarcinoma FFPE samples, 51 of which were further used for MSn-based structure prediction. 29157724 2 93 theme N-glycan 338:345 arg1 patterns 347:354 N-glycan patterns 338:354 N-glycan patterns 338:354 Current methods rely on either serum or fresh tissue samples; however, N-glycan patterns may differ between serum and tissue, as the proteins of serum originate from a variety of tissues. 29157724 4 94 theme tissue 604:609 arg1 sections 611:618 formalin-fixed paraffin-embedded (FFPE) tissue sections 564:618 formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients 564:652 Here, we used a profiling method based on formalin-fixed paraffin-embedded (FFPE) tissue sections from lung adenocarcinoma patients. 29157724 7 95 theme type 875:878 arg1 N-glycans 880:888 high mannose type N-glycans 862:888 high mannose type N-glycans 862:888 We show that high mannose type N-glycans are upregulated, while sialylated N-glycans are downregulated in our FFPE lung adenocarcinoma samples, compared to the control samples. 28978918 3 0 theme glycoforms 645:654 arg1 series 591:596 a series 589:596 a series of sequentially truncated high-mannose IgG1 Fc glycoforms 589:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 2 1 theme 297-linked 309:318 arg1 glycans 320:326 asparagine(N)297-linked glycans 296:326 asparagine(N)297-linked glycans 296:326 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 0 2 theme Fc 66:67 arg1 Structure 35:43 Structure 35:43 Structure 35:43 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 0 2 theme Fc 66:67 arg1 Dynamics 49:56 Dynamics 49:56 Dynamics 49:56 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 3 3 theme high-mannose 624:635 arg1 glycoforms 645:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 4 theme series 591:596 arg1 simulations 574:584 atomistic molecular dynamics simulations 545:584 atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms 545:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 5 theme N-glycan 747:754 arg1 composition 756:766 N-glycan composition 747:766 N-glycan composition 747:766 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 0 6 from Effects 0:6 arg1 Structure 35:43 Structure 35:43 Structure 35:43 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 0 6 from Effects 0:6 arg1 Dynamics 49:56 Dynamics 49:56 Dynamics 49:56 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 4 7 dep competent 880:888 arg1 compared 911:918 compared 911:918 compared to the truncated glycoforms 911:946 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 4 8 theme truncated 927:935 arg1 glycoforms 937:946 the truncated glycoforms 923:946 the truncated glycoforms 923:946 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 2 9 theme termini 455:461 arg1 distal 474:479 distal 474:479 distal 474:479 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 2 9 theme termini 455:461 arg1 modifications 429:441 the structural modifications 414:441 the structural modifications 414:441 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 2 10 link 297-linked 309:318 arg1 glycans 320:326 asparagine(N)297-linked glycans 296:326 asparagine(N)297-linked glycans 296:326 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 4 11 theme High-mannose 806:817 arg1 glycoform 819:827 High-mannose glycoform 806:827 High-mannose glycoform 806:827 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 1 12 theme monoclonal 230:239 arg1 antibodies 241:250 monoclonal antibodies 230:250 monoclonal antibodies 230:250 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 2 13 theme N-glycan 446:453 arg1 termini 455:461 N-glycan termini 446:461 N-glycan termini 446:461 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 4 14 with interface 1005:1013 arg1 receptor 1027:1034 the Fc receptor 1020:1034 the Fc receptor 1020:1034 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 2 15 theme Fc 370:371 arg1 affinity 353:360 the binding affinity 341:360 the binding affinity of IgG1 Fc to Fc γ receptors 341:389 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 2 16 from interface 498:506 arg1 distal 474:479 distal 474:479 distal 474:479 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 2 16 from interface 498:506 arg1 modifications 429:441 the structural modifications 414:441 the structural modifications 414:441 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 5 17 theme trajectory 1063:1072 arg1 analyses 1074:1081 The trajectory analyses 1059:1081 The trajectory analyses 1059:1081 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 2 18 theme binding 345:351 arg1 affinity 353:360 the binding affinity 341:360 the binding affinity of IgG1 Fc to Fc γ receptors 341:389 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 4 19 theme IgG1 970:973 arg1 Fc 975:976 IgG1 Fc 970:976 IgG1 Fc N-glycan 970:985 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 3 20 theme C' 675:676 arg1 dynamic 723:729 dynamic 723:729 dynamic 723:729 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 20 theme C' 675:676 arg1 loop 679:682 the C'E loop 671:682 the C'E loop 671:682 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 5 21 theme glycan 1258:1263 arg1 termini 1265:1271 the glycan termini 1254:1271 the glycan termini 1254:1271 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 3 22 theme E 677:677 arg1 dynamic 723:729 dynamic 723:729 dynamic 723:729 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 22 theme E 677:677 arg1 loop 679:682 the C'E loop 671:682 the C'E loop 671:682 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 4 23 theme efficient 1040:1048 arg1 binding 1050:1056 efficient binding 1040:1056 efficient binding 1040:1056 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 1 24 theme human 158:162 arg1 antibodies 170:179 human serum antibodies 158:179 human serum antibodies 158:179 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 0 25 theme Composition 20:30 arg1 Implications 79:90 Their Implications 73:90 Their Implications for Antibody Engineering 73:115 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 0 25 theme Composition 20:30 arg1 Effects 0:6 Effects 0:6 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc 0:67 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 5 26 theme large 1117:1121 arg1 motions 1133:1139 large amplitude motions 1117:1139 large amplitude motions 1117:1139 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 5 27 theme termini 1265:1271 arg1 modification 1238:1249 enzymatic modification 1228:1249 enzymatic modification of the glycan termini 1228:1271 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 1 28 theme serum 164:168 arg1 antibodies 170:179 human serum antibodies 158:179 human serum antibodies 158:179 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 2 29 theme Fc 376:377 arg1 receptors 381:389 Fc γ receptors 376:389 Fc γ receptors 376:389 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 5 30 theme amplitude 1123:1131 arg1 motions 1133:1139 large amplitude motions 1117:1139 large amplitude motions 1117:1139 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 3 31 from changes 736:742 arg1 composition 756:766 N-glycan composition 747:766 N-glycan composition 747:766 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 1 32 theme antibodies 170:179 arg1 G1 133:134 Immunoglobulin G1 118:134 Immunoglobulin G1 (IgG1) 118:141 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 32 theme antibodies 170:179 arg1 subclass 146:153 a subclass 144:153 a subclass of human serum antibodies 144:179 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 33 theme human 261:265 arg1 diseases 267:274 human diseases 261:274 human diseases 261:274 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 4 34 theme FcγRIIIa 893:900 arg1 binding 902:908 FcγRIIIa binding 893:908 FcγRIIIa binding 893:908 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 0 35 theme Antibody 96:103 arg1 Engineering 105:115 Antibody Engineering 96:115 Antibody Engineering 96:115 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 4 36 theme N-glycan 978:985 arg1 role 962:965 a role 960:965 a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding 960:1056 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 2 37 theme IgG1 365:368 arg1 Fc 370:371 IgG1 Fc 365:371 IgG1 Fc 365:371 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 3 38 gly glycoforms 645:654 arg1 Fc 642:643 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 38 gly glycoforms 645:654 arg1 high-mannose 624:635 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 38 gly glycoforms 645:654 arg1 IgG1 637:640 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 5 39 contain has 1113:1115 arg2 motions 1133:1139 large amplitude motions 1117:1139 large amplitude motions 1117:1139 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 5 39 contain has 1113:1115 arg1 N-glycan 1104:1111 the N-glycan 1100:1111 the N-glycan 1100:1111 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 3 40 theme dynamics 565:572 arg1 simulations 574:584 atomistic molecular dynamics simulations 545:584 atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms 545:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 2 41 theme γ 379:379 arg1 receptors 381:389 Fc γ receptors 376:389 Fc γ receptors 376:389 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 3 42 theme Cγ2-Cγ3 692:698 arg1 dynamic 723:729 dynamic 723:729 dynamic 723:729 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 42 theme Cγ2-Cγ3 692:698 arg1 orientation 700:710 the Cγ2-Cγ3 orientation 688:710 the Cγ2-Cγ3 orientation 688:710 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 43 theme IgG1 637:640 arg1 glycoforms 645:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 5 44 theme enzymatic 1228:1236 arg1 modification 1238:1249 enzymatic modification 1228:1249 enzymatic modification of the glycan termini 1228:1271 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 2 45 theme binding 490:496 arg1 interface 498:506 the binding interface 486:506 the binding interface 486:506 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 3 46 theme truncated 614:622 arg1 glycoforms 645:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 3 47 theme atomistic 545:553 arg1 simulations 574:584 atomistic molecular dynamics simulations 545:584 atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms 545:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 1 48 theme Immunoglobulin 118:131 arg1 IgG1 137:140 IgG1 137:140 IgG1 137:140 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 48 theme Immunoglobulin 118:131 arg1 G1 133:134 Immunoglobulin G1 118:134 Immunoglobulin G1 (IgG1) 118:141 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 48 theme Immunoglobulin 118:131 arg1 subclass 146:153 a subclass 144:153 a subclass of human serum antibodies 144:179 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 48 theme Immunoglobulin 118:131 arg1 scaffold 206:213 the most widely used scaffold 185:213 the most widely used scaffold for developing monoclonal antibodies to treat human diseases 185:274 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 3 49 theme conformational 780:793 arg1 ensembles 795:803 their conformational ensembles 774:803 their conformational ensembles 774:803 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 0 50 from Implications 79:90 arg1 Structure 35:43 Structure 35:43 Structure 35:43 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 0 50 from Implications 79:90 arg1 Dynamics 49:56 Dynamics 49:56 Dynamics 49:56 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 3 51 theme molecular 555:563 arg1 simulations 574:584 atomistic molecular dynamics simulations 545:584 atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms 545:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 4 52 theme Fc 975:976 arg1 N-glycan 978:985 IgG1 Fc N-glycan 970:985 IgG1 Fc N-glycan 970:985 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 2 53 theme asparagine 296:305 arg1 glycans 320:326 asparagine(N)297-linked glycans 296:326 asparagine(N)297-linked glycans 296:326 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 0 54 theme IgG1 61:64 arg1 Fc 66:67 IgG1 Fc 61:67 IgG1 Fc 61:67 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 4 55 theme Fc 1024:1025 arg1 receptor 1027:1034 the Fc receptor 1020:1034 the Fc receptor 1020:1034 High-mannose glycoform preferentially samples conformations that are more competent to FcγRIIIa binding, compared to the truncated glycoforms, suggesting a role of IgG1 Fc N-glycan in optimizing the interface with the Fc receptor for efficient binding. 28978918 5 56 theme Fc-bound 1191:1198 arg1 forms 1212:1216 Fc-bound and unbound forms 1191:1216 Fc-bound and unbound forms 1191:1216 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 2 57 theme structural 418:427 arg1 distal 474:479 distal 474:479 distal 474:479 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 2 57 theme structural 418:427 arg1 modifications 429:441 the structural modifications 414:441 the structural modifications 414:441 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 3 58 theme Fc 642:643 arg1 glycoforms 645:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 sequentially truncated high-mannose IgG1 Fc glycoforms 601:654 Through atomistic molecular dynamics simulations of a series of sequentially truncated high-mannose IgG1 Fc glycoforms, we found that the C'E loop and the Cγ2-Cγ3 orientation are highly dynamic, and changes in N-glycan composition alter their conformational ensembles. 28978918 0 59 theme N-Glycan 11:18 arg1 Composition 20:30 N-Glycan Composition 11:30 N-Glycan Composition 11:30 Effects of N-Glycan Composition on Structure and Dynamics of IgG1 Fc and Their Implications for Antibody Engineering. 28978918 5 60 theme carbohydrate 1149:1160 arg1 moiety 1162:1167 the carbohydrate moiety 1145:1167 the carbohydrate moiety 1145:1167 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 2 61 theme glycans 320:326 arg1 composition 281:291 The composition 277:291 The composition of asparagine(N)297-linked glycans 277:326 The composition of asparagine(N)297-linked glycans can modulate the binding affinity of IgG1 Fc to Fc γ receptors, but it is unclear how the structural modifications of N-glycan termini, which are distal from the binding interface, contribute to the affinity. 28978918 5 62 theme unbound 1204:1210 arg1 forms 1212:1216 Fc-bound and unbound forms 1191:1216 Fc-bound and unbound forms 1191:1216 The trajectory analyses also reveal that the N-glycan has large amplitude motions and the carbohydrate moiety interconverts between Fc-bound and unbound forms, enabling enzymatic modification of the glycan termini. 28978918 1 63 theme used 201:204 arg1 G1 133:134 Immunoglobulin G1 118:134 Immunoglobulin G1 (IgG1) 118:141 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28978918 1 63 theme used 201:204 arg1 scaffold 206:213 the most widely used scaffold 185:213 the most widely used scaffold for developing monoclonal antibodies to treat human diseases 185:274 Immunoglobulin G1 (IgG1), a subclass of human serum antibodies, is the most widely used scaffold for developing monoclonal antibodies to treat human diseases. 28426178 11 0 theme N-glycan 1830:1837 arg1 elucidation 1849:1859 N-glycan structure elucidation 1830:1859 N-glycan structure elucidation 1830:1859 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 11 1 theme sample 1875:1880 arg1 injection 1882:1890 only one sample injection 1866:1890 only one sample injection 1866:1890 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 3 2 theme N-glycosylation 337:351 arg1 mandatory 356:364 mandatory 356:364 mandatory 356:364 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 2 theme N-glycosylation 337:351 arg1 characterization 317:332 The detailed characterization 304:332 The detailed characterization of N-glycosylation 304:351 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 3 dep driven 394:399 arg1 nontemplate 382:392 nontemplate 382:392 nontemplate 382:392 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 1 4 from importance 145:154 arg1 research 220:227 biological research 209:227 biological research 209:227 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 9 5 theme Exoglycosidase 1373:1386 arg1 studies 1388:1394 Exoglycosidase studies 1373:1394 Exoglycosidase studies 1373:1394 Exoglycosidase studies were carried out using neuramididase, galactosidase, and fucosidase to confirm the results of three dye cross-validation. 28426178 10 6 theme bead 1649:1652 arg1 preparation 1663:1673 magnetic bead assisted preparation 1640:1673 magnetic bead assisted preparation 1640:1673 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 10 7 theme separation 1603:1612 arg1 capacity 1572:1579 the parallel separation capacity 1548:1579 the parallel separation capacity of multiple-capillary separation 1548:1612 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 10 8 theme N-glycans 1745:1753 arg1 analysis 1733:1740 rapid and accurate analysis 1714:1740 rapid and accurate analysis of N-glycans 1714:1753 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 6 9 theme preparation 972:982 arg1 procedure 984:992 the sample preparation procedure 961:992 the sample preparation procedure 961:992 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 10 10 theme multiple-capillary 1584:1601 arg1 separation 1603:1612 multiple-capillary separation 1584:1612 multiple-capillary separation 1584:1612 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 6 11 theme purification 927:938 arg1 method 940:945 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 2 12 theme drugs 297:301 arg1 development 271:281 the development 267:281 the development of biosimilar drugs 267:301 Glycan profiling is also critical to the development of biosimilar drugs. 28426178 4 13 from difficulties 646:657 arg1 procedures 731:740 time-consuming sample pretreatment procedures 696:740 time-consuming sample pretreatment procedures 696:740 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 4 13 from difficulties 646:657 arg1 complexity 681:690 the structure complexity 667:690 the structure complexity 667:690 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 6 14 theme sample 965:970 arg1 procedure 984:992 the sample preparation procedure 961:992 the sample preparation procedure 961:992 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 1 15 theme N-glycan 60:67 arg1 structures 69:78 N-glycan structures 60:78 N-glycan structures 60:78 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 6 16 theme DNA 874:876 arg1 analyzer 878:885 a 16-capillary DNA analyzer 859:885 a 16-capillary DNA analyzer 859:885 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 2 17 theme biosimilar 286:295 arg1 drugs 297:301 biosimilar drugs 286:301 biosimilar drugs 286:301 Glycan profiling is also critical to the development of biosimilar drugs. 28426178 9 18 theme dye 1496:1498 arg1 cross-validation 1500:1515 three dye cross-validation 1490:1515 three dye cross-validation 1490:1515 Exoglycosidase studies were carried out using neuramididase, galactosidase, and fucosidase to confirm the results of three dye cross-validation. 28426178 8 19 theme fluorescent 1234:1244 arg1 dyes 1246:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 6 20 theme 16-capillary 861:872 arg1 analyzer 878:885 a 16-capillary DNA analyzer 859:885 a 16-capillary DNA analyzer 859:885 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 1 21 theme structures 69:78 arg1 Analysis 48:55 Analysis 48:55 Analysis of N-glycan structures 48:78 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 1 22 theme biopharma-based 159:173 arg1 applications 175:186 biopharma-based applications 159:186 biopharma-based applications 159:186 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 5 23 theme high 816:819 arg1 efficiency 832:841 its high separation efficiency 812:841 its high separation efficiency 812:841 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 11 24 theme structure 1839:1847 arg1 elucidation 1849:1859 N-glycan structure elucidation 1830:1859 N-glycan structure elucidation 1830:1859 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 7 25 theme database 1208:1215 arg1 entries 1217:1223 database entries 1208:1223 database entries 1208:1223 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 8 26 theme preparation 1354:1364 arg1 steps 1366:1370 sample preparation steps 1347:1370 sample preparation steps 1347:1370 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 0 27 theme N-glycan 13:20 arg1 analysis 22:29 N-glycan analysis 13:29 N-glycan analysis 13:29 Multiplexing N-glycan analysis by DNA analyzer. 28426178 7 28 theme migration 1187:1195 arg1 times 1197:1201 migration times 1187:1201 migration times 1187:1201 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 10 29 theme separation 1561:1570 arg1 capacity 1572:1579 the parallel separation capacity 1548:1579 the parallel separation capacity of multiple-capillary separation 1548:1612 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 10 30 theme exoglycosidase 1680:1693 arg1 treatment 1695:1703 exoglycosidase treatment 1680:1703 exoglycosidase treatment 1680:1703 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 7 31 theme identification 1147:1160 arg1 method 1162:1167 the typical identification method 1135:1167 the typical identification method 1135:1167 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 9 32 theme cross-validation 1500:1515 arg1 results 1479:1485 the results 1475:1485 the results of three dye cross-validation 1475:1515 Exoglycosidase studies were carried out using neuramididase, galactosidase, and fucosidase to confirm the results of three dye cross-validation. 28426178 1 33 theme growing 192:198 arg1 roles 200:204 growing roles 192:204 growing roles in biological research 192:227 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 8 34 theme identification 1310:1323 arg1 precision 1325:1333 the glycan identification precision 1299:1333 the glycan identification precision 1299:1333 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 10 35 theme parallel 1552:1559 arg1 capacity 1572:1579 the parallel separation capacity 1548:1579 the parallel separation capacity of multiple-capillary separation 1548:1612 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 5 36 theme techniques 772:781 arg1 one 753:755 one 753:755 one 753:755 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 5 36 theme techniques 772:781 arg1 techniques 772:781 the typical techniques 760:781 the typical techniques for N-glycan analysis due to its high separation efficiency 760:841 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 4 37 theme time-consuming 696:709 arg1 procedures 731:740 time-consuming sample pretreatment procedures 696:740 time-consuming sample pretreatment procedures 696:740 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 3 38 theme driven 394:399 arg1 process 401:407 a nontemplate driven process 380:407 a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity 380:497 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 38 theme driven 394:399 arg1 it 374:375 it 374:375 it 374:375 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 10 39 theme assisted 1654:1661 arg1 preparation 1663:1673 magnetic bead assisted preparation 1640:1673 magnetic bead assisted preparation 1640:1673 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 10 40 theme rapid 1714:1718 arg1 analysis 1733:1740 rapid and accurate analysis 1714:1740 rapid and accurate analysis of N-glycans 1714:1753 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 5 41 theme N-glycan 787:794 arg1 analysis 796:803 N-glycan analysis 787:803 N-glycan analysis due to its high separation efficiency 787:841 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 4 42 theme pretreatment 718:729 arg1 procedures 731:740 time-consuming sample pretreatment procedures 696:740 time-consuming sample pretreatment procedures 696:740 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 1 43 theme biological 209:218 arg1 research 220:227 biological research 209:227 biological research 209:227 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 0 44 theme DNA 34:36 arg1 analyzer 38:45 DNA analyzer 34:45 DNA analyzer 34:45 Multiplexing N-glycan analysis by DNA analyzer. 28426178 3 45 theme critical 443:450 arg1 bio-activity 486:497 bio-activity 486:497 bio-activity 486:497 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 45 theme critical 443:450 arg1 properties 452:461 critical properties 443:461 critical properties such as bio-safety and bio-activity 443:497 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 45 theme critical 443:450 arg1 bio-safety 471:480 bio-safety 471:480 bio-safety 471:480 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 6 46 theme glycan 920:925 arg1 method 940:945 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 7 47 theme 8-aminopyrene-1,3,6-trisulfonic 1091:1121 arg1 dye 1068:1070 the labeling dye 1055:1070 the labeling dye used for CE-LIF 1055:1086 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 7 47 theme 8-aminopyrene-1,3,6-trisulfonic 1091:1121 arg1 acid 1123:1126 8-aminopyrene-1,3,6-trisulfonic acid 1091:1126 8-aminopyrene-1,3,6-trisulfonic acid 1091:1126 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 4 48 theme sample 711:716 arg1 procedures 731:740 time-consuming sample pretreatment procedures 696:740 time-consuming sample pretreatment procedures 696:740 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 8 49 used used 1256:1259 arg2 dyes 1246:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 4 50 theme N-glycans 571:579 arg1 mixtures 559:566 highly complex mixtures 544:566 highly complex mixtures of N-glycans 544:579 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 3 51 theme detailed 308:315 arg1 mandatory 356:364 mandatory 356:364 mandatory 356:364 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 3 51 theme detailed 308:315 arg1 characterization 317:332 The detailed characterization 304:332 The detailed characterization of N-glycosylation 304:351 The detailed characterization of N-glycosylation is mandatory because it is a nontemplate driven process and that significantly influences critical properties such as bio-safety and bio-activity. 28426178 6 52 theme bead 915:918 arg1 method 940:945 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 2 53 theme Glycan 230:235 arg1 profiling 237:245 Glycan profiling 230:245 Glycan profiling 230:245 Glycan profiling is also critical to the development of biosimilar drugs. 28426178 10 54 theme magnetic 1640:1647 arg1 preparation 1663:1673 magnetic bead assisted preparation 1640:1673 magnetic bead assisted preparation 1640:1673 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 6 55 theme N-glycan 1017:1024 arg1 throughput 1032:1041 N-glycan assay throughput 1017:1041 N-glycan assay throughput 1017:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 7 56 theme typical 1139:1145 arg1 method 1162:1167 the typical identification method 1135:1167 the typical identification method 1135:1167 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 6 57 theme magnetic 906:913 arg1 method 940:945 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput 904:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 1 58 from roles 200:204 arg1 research 220:227 biological research 209:227 biological research 209:227 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 10 59 theme accurate 1724:1731 arg1 analysis 1733:1740 rapid and accurate analysis 1714:1740 rapid and accurate analysis of N-glycans 1714:1753 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 10 60 theme labeling 1625:1632 arg1 dyes 1634:1637 three labeling dyes 1619:1637 three labeling dyes 1619:1637 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 8 61 theme new 1230:1232 arg1 dyes 1246:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes 1226:1249 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 1 62 theme due 123:125 arg1 years 117:121 the years 113:121 the years due to their critical importance to biopharma-based applications and growing roles in biological research 113:227 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 5 63 theme typical 764:770 arg1 techniques 772:781 the typical techniques 760:781 the typical techniques for N-glycan analysis due to its high separation efficiency 760:841 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 11 64 theme new 1762:1764 arg1 methods 1766:1772 These new methods 1756:1772 These new methods 1756:1772 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 11 65 theme useful 1790:1795 arg1 information 1808:1818 enough useful structural information 1783:1818 enough useful structural information to permit N-glycan structure elucidation with only one sample injection 1783:1890 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 5 66 theme separation 821:830 arg1 efficiency 832:841 its high separation efficiency 812:841 its high separation efficiency 812:841 CE-LIF is one of the typical techniques for N-glycan analysis due to its high separation efficiency. 28426178 11 67 theme structural 1797:1806 arg1 information 1808:1818 enough useful structural information 1783:1818 enough useful structural information to permit N-glycan structure elucidation with only one sample injection 1783:1890 These new methods provided enough useful structural information to permit N-glycan structure elucidation with only one sample injection. 28426178 10 68 theme optimized 1522:1530 arg1 method 1532:1537 The optimized method 1518:1537 The optimized method 1518:1537 The optimized method combines the parallel separation capacity of multiple-capillary separation with three labeling dyes, magnetic bead assisted preparation, and exoglycosidase treatment to allow rapid and accurate analysis of N-glycans. 28426178 8 69 theme sample 1347:1352 arg1 steps 1366:1370 sample preparation steps 1347:1370 sample preparation steps 1347:1370 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 4 70 theme complex 551:557 arg1 mixtures 559:566 highly complex mixtures 544:566 highly complex mixtures of N-glycans 544:579 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28426178 8 71 theme glycan 1303:1308 arg1 precision 1325:1333 the glycan identification precision 1299:1333 the glycan identification precision 1299:1333 Two new fluorescent dyes were used to either cross-validate and increase the glycan identification precision or simplify sample preparation steps. 28426178 7 72 theme labeling 1059:1066 arg1 dye 1068:1070 the labeling dye 1055:1070 the labeling dye used for CE-LIF 1055:1086 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 7 72 theme labeling 1059:1066 arg1 acid 1123:1126 8-aminopyrene-1,3,6-trisulfonic acid 1091:1126 8-aminopyrene-1,3,6-trisulfonic acid 1091:1126 Routinely, the labeling dye used for CE-LIF is 8-aminopyrene-1,3,6-trisulfonic acid, while the typical identification method involves matching migration times with database entries. 28426178 1 73 theme critical 136:143 arg1 importance 145:154 their critical importance 130:154 their critical importance to biopharma-based applications 130:186 Analysis of N-glycan structures has been gaining attentions over the years due to their critical importance to biopharma-based applications and growing roles in biological research. 28426178 6 74 theme assay 1026:1030 arg1 throughput 1032:1041 N-glycan assay throughput 1017:1041 N-glycan assay throughput 1017:1041 In this paper, a 16-capillary DNA analyzer was coupled with a magnetic bead glycan purification method to accelerate the sample preparation procedure and therefore increase N-glycan assay throughput. 28426178 4 75 theme structure 671:679 arg1 complexity 681:690 the structure complexity 667:690 the structure complexity 667:690 The ability to comprehensively characterize highly complex mixtures of N-glycans has been analytically challenging and stimulating because of the difficulties in both the structure complexity and time-consuming sample pretreatment procedures. 28489544 5 0 theme glycopeptides 808:820 arg1 MS/MS 799:803 HCD MS/MS 795:803 HCD MS/MS of glycopeptides 795:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 4 1 theme input 532:536 arg1 spectra 538:544 the input spectra 528:544 the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme 528:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 6 2 contain has 905:907 arg2 performance 916:926 better performance 909:926 better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 909:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 6 2 contain has 905:907 arg1 GlycoNovoDB 856:866 GlycoNovoDB 856:866 GlycoNovoDB 856:866 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 5 3 theme software 716:723 arg1 approach 687:694 The proposed approach 674:694 The proposed approach 674:694 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 5 3 theme software 716:723 arg1 GlycoNovoDB 731:741 GlycoNovoDB 731:741 GlycoNovoDB 731:741 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 5 3 theme software 716:723 arg1 tool 725:728 a software tool 714:728 a software tool 714:728 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 0 4 theme MS/MS 75:79 arg1 Spectra 81:87 HCD MS/MS Spectra 71:87 HCD MS/MS Spectra 71:87 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 4 5 theme glycan 551:556 arg1 structures 558:567 glycan structures 551:567 glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme 551:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 3 6 theme mass 407:410 arg1 spectra 412:418 mass spectra 407:418 mass spectra 407:418 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 6 7 theme sequencing 965:974 arg1 algorithm 976:984 our previously proposed de novo sequencing algorithm 933:984 our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 933:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 4 8 theme de 630:631 arg1 scheme 666:671 a de novo sequencing assisted ranking scheme 628:671 a de novo sequencing assisted ranking scheme 628:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 2 9 theme oligosaccharides 295:310 arg1 structures 281:290 structures 281:290 structures of oligosaccharides 281:310 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 5 10 theme HCD 795:797 arg1 MS/MS 799:803 HCD MS/MS 795:803 HCD MS/MS of glycopeptides 795:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 6 11 theme de 957:958 arg1 algorithm 976:984 our previously proposed de novo sequencing algorithm 933:984 our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 933:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 0 12 theme Glycan 34:39 arg1 Structure 41:49 N-Linked Glycan Structure 25:49 N-Linked Glycan Structure 25:49 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 5 13 gly glycopeptides 808:820 arg2 glycopeptides 808:820 glycopeptides 808:820 glycopeptides 808:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 0 14 theme Improved 3:10 arg1 Approach 12:19 An Improved Approach 0:19 An Improved Approach for N-Linked Glycan Structure 0:49 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 6 15 theme proposed 948:955 arg1 algorithm 976:984 our previously proposed de novo sequencing algorithm 933:984 our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 933:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 5 16 theme proposed 678:685 arg1 approach 687:694 The proposed approach 674:694 The proposed approach 674:694 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 5 16 theme proposed 678:685 arg1 tool 725:728 a software tool 714:728 a software tool 714:728 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 1 17 from modification 148:159 arg1 proteins 164:171 proteins 164:171 proteins 164:171 Glycosylation is a frequently observed post-translational modification on proteins. 28489544 3 18 theme great 431:435 arg1 challenge 437:445 a great challenge 429:445 a great challenge in glycoproteomics research 429:473 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 4 19 theme structure 592:600 arg1 database 602:609 a glycan structure database 583:609 a glycan structure database 583:609 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 4 20 dep de 630:631 arg1 novo 633:636 novo 633:636 novo 633:636 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 4 21 theme glycan 585:590 arg1 database 602:609 a glycan structure database 583:609 a glycan structure database 583:609 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 6 22 theme Experimental 823:834 arg1 results 836:842 Experimental results 823:842 Experimental results 823:842 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 0 23 from Spectra 81:87 arg1 Identification 51:64 Identification 51:64 Identification 51:64 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 3 24 theme glycoproteomics 450:464 arg1 research 466:473 glycoproteomics research 450:473 glycoproteomics research 450:473 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 5 25 theme automated 748:756 arg1 identification 775:788 automated glycan structure identification 748:788 automated glycan structure identification from HCD MS/MS of glycopeptides 748:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 2 26 theme mass 192:195 arg1 MS/MS 211:215 MS/MS 211:215 MS/MS 211:215 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 26 theme mass 192:195 arg1 spectrometry 197:208 tandem mass spectrometry 185:208 tandem mass spectrometry (MS/MS) 185:216 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 26 theme mass 192:195 arg1 technique 252:260 an efficient analytical technique 228:260 an efficient analytical technique for characterizing structures of oligosaccharides 228:310 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 27 theme analytical 241:250 arg1 spectrometry 197:208 tandem mass spectrometry 185:208 tandem mass spectrometry (MS/MS) 185:216 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 27 theme analytical 241:250 arg1 technique 252:260 an efficient analytical technique 228:260 an efficient analytical technique for characterizing structures of oligosaccharides 228:310 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 0 28 theme N-Linked 25:32 arg1 Structure 41:49 N-Linked Glycan Structure 25:49 N-Linked Glycan Structure 25:49 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 5 29 theme glycan 758:763 arg1 identification 775:788 automated glycan structure identification 748:788 automated glycan structure identification from HCD MS/MS of glycopeptides 748:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 2 30 theme tandem 185:190 arg1 MS/MS 211:215 MS/MS 211:215 MS/MS 211:215 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 30 theme tandem 185:190 arg1 spectrometry 197:208 tandem mass spectrometry 185:208 tandem mass spectrometry (MS/MS) 185:216 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 30 theme tandem 185:190 arg1 technique 252:260 an efficient analytical technique 228:260 an efficient analytical technique for characterizing structures of oligosaccharides 228:310 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 31 theme efficient 231:239 arg1 spectrometry 197:208 tandem mass spectrometry 185:208 tandem mass spectrometry (MS/MS) 185:216 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 2 31 theme efficient 231:239 arg1 technique 252:260 an efficient analytical technique 228:260 an efficient analytical technique for characterizing structures of oligosaccharides 228:310 Currently, tandem mass spectrometry (MS/MS) serves as an efficient analytical technique for characterizing structures of oligosaccharides. 28489544 5 32 theme structure 765:773 arg1 identification 775:788 automated glycan structure identification 748:788 automated glycan structure identification from HCD MS/MS of glycopeptides 748:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 3 33 theme effective 333:341 arg1 approaches 357:366 effective computational approaches 333:366 effective computational approaches 333:366 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 6 34 theme software 1005:1012 arg1 DB 1026:1027 another software GlycoMaster DB 997:1027 our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 933:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 3 35 theme computational 343:355 arg1 approaches 357:366 effective computational approaches 333:366 effective computational approaches 333:366 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 6 36 theme better 909:914 arg1 performance 916:926 better performance 909:926 better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 909:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 1 37 theme observed 120:127 arg1 Glycosylation 90:102 Glycosylation 90:102 Glycosylation 90:102 Glycosylation is a frequently observed post-translational modification on proteins. 28489544 1 37 theme observed 120:127 arg1 modification 148:159 a frequently observed post-translational modification 107:159 a frequently observed post-translational modification on proteins 107:171 Glycosylation is a frequently observed post-translational modification on proteins. 28489544 1 38 theme post-translational 129:146 arg1 Glycosylation 90:102 Glycosylation 90:102 Glycosylation 90:102 Glycosylation is a frequently observed post-translational modification on proteins. 28489544 1 38 theme post-translational 129:146 arg1 modification 148:159 a frequently observed post-translational modification 107:159 a frequently observed post-translational modification on proteins 107:171 Glycosylation is a frequently observed post-translational modification on proteins. 28489544 4 39 theme assisted 649:656 arg1 scheme 666:671 a de novo sequencing assisted ranking scheme 628:671 a de novo sequencing assisted ranking scheme 628:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 5 40 from MS/MS 799:803 arg1 identification 775:788 automated glycan structure identification 748:788 automated glycan structure identification from HCD MS/MS of glycopeptides 748:820 The proposed approach is implemented as a software tool, GlycoNovoDB, for automated glycan structure identification from HCD MS/MS of glycopeptides. 28489544 4 41 with spectra 538:544 arg1 structures 558:567 glycan structures 551:567 glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme 551:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 4 42 theme sequencing 638:647 arg1 scheme 666:671 a de novo sequencing assisted ranking scheme 628:671 a de novo sequencing assisted ranking scheme 628:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 28489544 0 43 theme HCD 71:73 arg1 Spectra 81:87 HCD MS/MS Spectra 71:87 HCD MS/MS Spectra 71:87 An Improved Approach for N-Linked Glycan Structure Identification from HCD MS/MS Spectra. 28489544 3 44 theme glycan 384:389 arg1 structures 391:400 glycan structures 384:400 glycan structures 384:400 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 3 45 from challenge 437:445 arg1 research 466:473 glycoproteomics research 450:473 glycoproteomics research 450:473 However, developing effective computational approaches for identifying glycan structures from mass spectra is still a great challenge in glycoproteomics research. 28489544 6 46 dep de 957:958 arg1 novo 960:963 novo 960:963 novo 960:963 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 6 47 theme GlycoMaster 1014:1024 arg1 DB 1026:1027 another software GlycoMaster DB 997:1027 our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB 933:1027 Experimental results showed that GlycoNovoDB can identify glycans effectively and has better performance than our previously proposed de novo sequencing algorithm as well as another software GlycoMaster DB. 28489544 4 48 theme ranking 658:664 arg1 scheme 666:671 a de novo sequencing assisted ranking scheme 628:671 a de novo sequencing assisted ranking scheme 628:671 In this study, we proposed an approach for matching the input spectra with glycan structures acquired from a glycan structure database by incorporating a de novo sequencing assisted ranking scheme. 24616211 3 0 link linked 487:492 arg1 hexasaccharide 494:507 the archaeal L-asparagine linked hexasaccharide 461:507 the archaeal L-asparagine linked hexasaccharide 461:507 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 0 1 link asparagine-linked 22:38 arg1 N-glycan 49:56 asparagine-linked archaeal N-glycan 22:56 asparagine-linked archaeal N-glycan 22:56 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 4 2 theme late 769:772 arg1 stage 774:778 late stage 769:778 late stage 769:778 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 2 3 theme N-glycans 308:316 arg1 synthesis 282:290 Chemical synthesis 273:290 Chemical synthesis of the archaeal N-glycans 273:316 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 4 4 theme orthogonal 739:748 arg1 glycosylations 750:763 orthogonal glycosylations 739:763 orthogonal glycosylations 739:763 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 1 5 link N-linked 96:103 arg1 glycoproteins 105:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 2 6 theme archaeal 299:306 arg1 N-glycans 308:316 the archaeal N-glycans 295:316 the archaeal N-glycans 295:316 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 4 7 theme protected 695:703 arg1 blocks 728:733 regioselectively protected thioglycoside building blocks 678:733 regioselectively protected thioglycoside building blocks 678:733 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 1 8 theme Several 88:94 arg1 glycoproteins 105:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 3 9 theme archaeal 465:472 arg1 hexasaccharide 494:507 the archaeal L-asparagine linked hexasaccharide 461:507 the archaeal L-asparagine linked hexasaccharide 461:507 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 3 10 theme total 442:446 arg1 synthesis 448:456 the first total synthesis 432:456 the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus 432:536 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 3 11 from fervidus 529:536 arg1 synthesis 448:456 the first total synthesis 432:456 the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus 432:536 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 1 12 theme N-linked 96:103 arg1 glycoproteins 105:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 3 13 theme L-asparagine 474:485 arg1 hexasaccharide 494:507 the archaeal L-asparagine linked hexasaccharide 461:507 the archaeal L-asparagine linked hexasaccharide 461:507 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 3 14 theme [3+3 576:579 arg1 approach 596:603 a highly convergent [3+3] glycosylation approach 556:603 a highly convergent [3+3] glycosylation approach 556:603 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 2 15 from function 379:386 arg1 archaea 416:422 archaea 416:422 archaea 416:422 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 0 16 theme Chemical 0:7 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus 0:85 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 2 17 theme crucial 331:337 arg1 step 339:342 a crucial step 329:342 a crucial step towards understanding the putative function of protein glycosylation in archaea 329:422 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 0 18 theme asparagine-linked 22:38 arg1 N-glycan 49:56 asparagine-linked archaeal N-glycan 22:56 asparagine-linked archaeal N-glycan 22:56 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 3 19 theme high 608:611 arg1 yields 621:626 high overall yields 608:626 high overall yields 608:626 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 4 20 theme thioglycoside 705:717 arg1 blocks 728:733 regioselectively protected thioglycoside building blocks 678:733 regioselectively protected thioglycoside building blocks 678:733 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 1 21 gly glycoproteins 105:117 arg1 glycoproteins 105:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins 88:117 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 3 22 theme overall 613:619 arg1 yields 621:626 high overall yields 608:626 high overall yields 608:626 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 3 23 theme convergent 565:574 arg1 approach 596:603 a highly convergent [3+3] glycosylation approach 556:603 a highly convergent [3+3] glycosylation approach 556:603 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 0 24 theme N-glycan 49:56 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus 0:85 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 4 25 theme blocks 728:733 arg1 preparation 663:673 efficient preparation 653:673 efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations 653:763 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 4 25 theme blocks 728:733 arg1 stage 774:778 late stage 769:778 late stage 769:778 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 3 26 theme linked 487:492 arg1 hexasaccharide 494:507 the archaeal L-asparagine linked hexasaccharide 461:507 the archaeal L-asparagine linked hexasaccharide 461:507 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 0 27 theme archaeal 40:47 arg1 N-glycan 49:56 asparagine-linked archaeal N-glycan 22:56 asparagine-linked archaeal N-glycan 22:56 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 3 28 theme hexasaccharide 494:507 arg1 synthesis 448:456 the first total synthesis 432:456 the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus 432:536 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 0 29 from fervidus 78:85 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus 0:85 Chemical synthesis of asparagine-linked archaeal N-glycan from Methanothermus fervidus. 24616211 2 30 theme putative 370:377 arg1 function 379:386 the putative function 366:386 the putative function of protein glycosylation in archaea 366:422 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 3 31 theme first 436:440 arg1 synthesis 448:456 the first total synthesis 432:456 the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus 432:536 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 1 32 from survival 214:221 arg1 conditions 261:270 extreme conditions 253:270 extreme conditions 253:270 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 3 33 theme glycosylation 582:594 arg1 approach 596:603 a highly convergent [3+3] glycosylation approach 556:603 a highly convergent [3+3] glycosylation approach 556:603 Herein the first total synthesis of the archaeal L-asparagine linked hexasaccharide from Methanothermus fervidus is reported using a highly convergent [3+3] glycosylation approach in high overall yields. 24616211 4 34 dep preparation 663:673 arg1 N-aspartylation 780:794 N-aspartylation 780:794 N-aspartylation 780:794 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 2 35 theme Chemical 273:280 arg1 synthesis 282:290 Chemical synthesis 273:290 Chemical synthesis of the archaeal N-glycans 273:316 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 1 36 theme growing 164:170 arg1 evidence 172:179 growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions 164:270 growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions 164:270 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 1 37 theme archaea 242:248 arg1 survival 214:221 survival 214:221 survival 214:221 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 24616211 4 38 theme building 719:726 arg1 blocks 728:733 regioselectively protected thioglycoside building blocks 678:733 regioselectively protected thioglycoside building blocks 678:733 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 2 39 theme glycosylation 399:411 arg1 function 379:386 the putative function 366:386 the putative function of protein glycosylation in archaea 366:422 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 4 40 theme efficient 653:661 arg1 preparation 663:673 efficient preparation 653:673 efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations 653:763 The synthesis relies on efficient preparation of regioselectively protected thioglycoside building blocks for orthogonal glycosylations and late stage N-aspartylation. 24616211 2 41 theme protein 391:397 arg1 glycosylation 399:411 protein glycosylation 391:411 protein glycosylation 391:411 Chemical synthesis of the archaeal N-glycans represents a crucial step towards understanding the putative function of protein glycosylation in archaea. 24616211 1 42 theme extreme 253:259 arg1 conditions 261:270 extreme conditions 253:270 extreme conditions 253:270 Several N-linked glycoproteins have been identified in archaea and there is growing evidence that the N-glycan is involved in survival and functioning of archaea in extreme conditions. 26571101 2 0 gly glycoprotein 348:359 arg1 glycoprotein 348:359 current glycoprotein analyses 340:368 current glycoprotein analyses 340:368 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 1 theme total 798:802 arg1 glycan 813:818 the total N-linked glycan 794:818 the total N-linked glycan 794:818 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 4 2 theme total 984:988 arg1 occupancies 1015:1025 total and site-specific glycan occupancies 984:1025 total and site-specific glycan occupancies 984:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 3 3 theme N-linked 564:571 arg1 glycans 573:579 N-linked glycans 564:579 N-linked glycans 564:579 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 4 gly glycosites 740:749 arg2 glycosites 740:749 individual glycosites 729:749 individual glycosites 729:749 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 0 5 theme glycans 86:92 arg1 extraction 63:72 solid-phase extraction 51:72 solid-phase extraction of N-linked glycans and glycosite-containing peptides 51:126 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 2 6 theme glycosylation 413:425 arg1 site 427:430 the de-glycosylated glycosylation site 393:430 the de-glycosylated glycosylation site 393:430 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 0 7 theme peptides 119:126 arg1 extraction 63:72 solid-phase extraction 51:72 solid-phase extraction of N-linked glycans and glycosite-containing peptides 51:126 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 4 8 theme site-specific 994:1006 arg1 occupancies 1015:1025 total and site-specific glycan occupancies 984:1025 total and site-specific glycan occupancies 984:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 2 9 theme current 340:346 arg1 analyses 361:368 current glycoprotein analyses 340:368 current glycoprotein analyses 340:368 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 2 10 theme glycoprotein 348:359 arg1 analyses 361:368 current glycoprotein analyses 340:368 current glycoprotein analyses 340:368 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 0 11 theme glycosite-containing 98:117 arg1 peptides 119:126 glycosite-containing peptides 98:126 glycosite-containing peptides 98:126 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 4 12 from alterations 969:979 arg1 occupancies 1015:1025 total and site-specific glycan occupancies 984:1025 total and site-specific glycan occupancies 984:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 4 13 theme NGAG 899:902 arg1 method 904:909 The NGAG method 895:909 The NGAG method 895:909 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 3 14 link N-linked 804:811 arg1 glycan 813:818 the total N-linked glycan 794:818 the total N-linked glycan 794:818 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 15 theme phase 544:548 arg1 extraction 550:559 solid phase extraction 538:559 solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) 538:620 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 16 gly glycoprotein 312:323 arg1 glycoprotein 312:323 glycoprotein conformations 312:337 glycoprotein conformations 312:337 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 17 gly glycoprotein 854:865 arg1 glycoprotein 854:865 glycoprotein content 854:873 glycoprotein content of complex samples 854:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 1 18 gly glycoproteins 245:257 arg1 glycoproteins 245:257 glycoproteins 245:257 glycoproteins 245:257 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 2 19 theme conformations 325:337 arg1 heterogeneity 295:307 heterogeneity 295:307 heterogeneity 295:307 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 2 19 theme conformations 325:337 arg1 complexity 280:289 complexity 280:289 complexity 280:289 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 20 dep providing 766:774 arg1 addition 754:761 addition 754:761 addition 754:761 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 21 theme N-linked 804:811 arg1 glycan 813:818 the total N-linked glycan 794:818 the total N-linked glycan 794:818 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 22 theme glycoprotein 312:323 arg1 conformations 325:337 glycoprotein conformations 312:337 glycoprotein conformations 312:337 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 4 23 gly glycoprotein 956:967 arg1 glycoprotein 956:967 glycoprotein alterations 956:979 glycoprotein alterations in total and site-specific glycan occupancies 956:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 2 24 dep complexity 280:289 arg1 the 276:278 the 276:278 the 276:278 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 2 25 gly heterogeneity 295:307 arg1 conformations 325:337 glycoprotein conformations 312:337 glycoprotein conformations 312:337 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 26 theme comprehensive 630:642 arg1 characterization 644:659 the comprehensive characterization 626:659 the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples 626:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 27 theme peptides 606:613 arg1 extraction 550:559 solid phase extraction 538:559 solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) 538:620 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 28 theme -containing 443:453 arg1 peptides 455:462 the de-glycosylated glycosylation site (glycosite)-containing peptides 393:462 the de-glycosylated glycosylation site (glycosite)-containing peptides 393:462 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 0 29 theme Comprehensive 0:12 arg1 analysis 14:21 Comprehensive analysis 0:21 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 0:127 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 3 30 theme glycoprotein 854:865 arg1 content 867:873 glycoprotein content 854:873 glycoprotein content of complex samples 854:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 4 31 theme glycoprotein 956:967 arg1 alterations 969:979 glycoprotein alterations 956:979 glycoprotein alterations in total and site-specific glycan occupancies 956:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 0 32 theme protein 26:32 arg1 glycosylation 34:46 protein glycosylation 26:46 protein glycosylation 26:46 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 3 33 link N-linked 564:571 arg1 glycans 573:579 N-linked glycans 564:579 N-linked glycans 564:579 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 34 theme glycoproteins 664:676 arg1 characterization 644:659 the comprehensive characterization 626:659 the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples 626:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 35 gly glycosite 433:441 arg2 glycosite 433:441 glycosite 433:441 glycosite 433:441 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 36 theme individual 729:738 arg1 glycosites 740:749 individual glycosites 729:749 individual glycosites 729:749 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 1 37 theme Comprehensive 129:141 arg1 characterization 143:158 Comprehensive characterization 129:158 Comprehensive characterization of protein glycosylation 129:183 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 1 38 theme glycoproteins 245:257 arg1 function 233:240 function 233:240 function 233:240 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 1 38 theme glycoproteins 245:257 arg1 structure 219:227 structure 219:227 structure 219:227 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 2 39 gly glycosylation 413:425 arg2 site 427:430 the de-glycosylated glycosylation site 393:430 the de-glycosylated glycosylation site 393:430 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 3 40 gly glycoproteins 664:676 arg1 glycoproteins 664:676 glycoproteins 664:676 glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples 664:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 41 theme released 471:478 arg1 glycans 480:486 the released glycans 467:486 the released glycans 467:486 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 0 42 theme glycosylation 34:46 arg1 analysis 14:21 Comprehensive analysis 0:21 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 0:127 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 3 43 theme samples 886:892 arg1 glycan 813:818 the total N-linked glycan 794:818 the total N-linked glycan 794:818 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 43 theme samples 886:892 arg1 peptide 842:848 glycosite-containing peptide 821:848 glycosite-containing peptide 821:848 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 43 theme samples 886:892 arg1 content 867:873 glycoprotein content 854:873 glycoprotein content of complex samples 854:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 44 theme chemoenzymatic 509:522 arg1 method 524:529 a chemoenzymatic method 507:529 a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples 507:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 45 theme de-glycosylated 397:411 arg1 site 427:430 the de-glycosylated glycosylation site 393:430 the de-glycosylated glycosylation site 393:430 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 0 46 theme solid-phase 51:61 arg1 extraction 63:72 solid-phase extraction 51:72 solid-phase extraction of N-linked glycans and glycosite-containing peptides 51:126 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 2 47 gly de-glycosylated 397:411 arg1 site 427:430 the de-glycosylated glycosylation site 393:430 the de-glycosylated glycosylation site 393:430 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 1 48 dep structure 219:227 arg1 the 215:217 the 215:217 the 215:217 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 0 49 link N-linked 77:84 arg1 glycans 86:92 N-linked glycans 77:92 N-linked glycans 77:92 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 3 50 theme complex 878:884 arg1 samples 886:892 complex samples 878:892 complex samples 878:892 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 51 theme glycosite-containing 821:840 arg1 peptide 842:848 glycosite-containing peptide 821:848 glycosite-containing peptide 821:848 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 52 theme glycans 573:579 arg1 extraction 550:559 solid phase extraction 538:559 solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) 538:620 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 2 53 dep -containing 443:453 arg1 site 427:430 the de-glycosylated glycosylation site 393:430 the de-glycosylated glycosylation site 393:430 However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. 26571101 1 54 theme protein 163:169 arg1 glycosylation 171:183 protein glycosylation 163:183 protein glycosylation 163:183 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 3 55 theme glycan 704:709 arg1 heterogeneity 711:723 glycan heterogeneity 704:723 glycan heterogeneity for individual glycosites 704:749 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 1 56 theme glycosylation 171:183 arg1 characterization 143:158 Comprehensive characterization 129:158 Comprehensive characterization of protein glycosylation 129:183 Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. 26571101 3 57 theme solid 538:542 arg1 extraction 550:559 solid phase extraction 538:559 solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) 538:620 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 0 58 theme N-linked 77:84 arg1 glycans 86:92 N-linked glycans 77:92 N-linked glycans 77:92 Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides. 26571101 4 59 theme glycan 1008:1013 arg1 occupancies 1015:1025 total and site-specific glycan occupancies 984:1025 total and site-specific glycan occupancies 984:1025 The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies. 26571101 3 60 theme glycosite-containing 585:604 arg1 NGAG 616:619 NGAG 616:619 NGAG 616:619 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 26571101 3 60 theme glycosite-containing 585:604 arg1 peptides 606:613 glycosite-containing peptides 585:613 glycosite-containing peptides (NGAG) 585:620 Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. 27993676 7 0 theme l-Gal 1128:1132 arg1 in vitro 1154:1161 l-Gal containing N-glycan in vitro 1128:1161 l-Gal containing N-glycan in vitro 1128:1161 Our results clearly demonstrate that MmFUT8 and AtFucTA transfer l-Gal residues from GDP-l-Gal and synthesize l-Gal containing N-glycan in vitro. 27993676 7 1 from GDP-l-Gal 1103:1111 arg1 residues 1089:1096 l-Gal residues 1083:1096 l-Gal residues from GDP-l-Gal 1083:1111 Our results clearly demonstrate that MmFUT8 and AtFucTA transfer l-Gal residues from GDP-l-Gal and synthesize l-Gal containing N-glycan in vitro. 27993676 1 2 located detected 148:155 arg2 l-Galactose 67:77 l-Galactose 67:77 l-Galactose (l-Gal) containing N-glycans 67:106 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 2 located detected 148:155 arg2 polysaccharides 122:136 cell wall polysaccharides 112:136 cell wall polysaccharides 112:136 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 2 located detected 148:155 arg1 mutant 185:190 the l-Fuc deficient mur1 mutant 160:190 the l-Fuc deficient mur1 mutant of Arabidopsis thaliana 160:214 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 2 located detected 148:155 arg2 l-Gal 80:84 l-Gal 80:84 l-Gal 80:84 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 3 theme deficient 170:178 arg1 mutant 185:190 the l-Fuc deficient mur1 mutant 160:190 the l-Fuc deficient mur1 mutant of Arabidopsis thaliana 160:214 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 7 4 theme l-Gal 1083:1087 arg1 residues 1089:1096 l-Gal residues 1083:1096 l-Gal residues from GDP-l-Gal 1083:1111 Our results clearly demonstrate that MmFUT8 and AtFucTA transfer l-Gal residues from GDP-l-Gal and synthesize l-Gal containing N-glycan in vitro. 27993676 4 5 theme α1,3-fucosyltransferase 631:653 arg1 assay 567:571 the l-galactosylation assay 545:571 the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA) 545:663 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 3 6 contain carried 415:421 arg3 study 405:409 this study 400:409 this study 400:409 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 3 6 contain carried 415:421 arg1 we 412:413 we 412:413 we 412:413 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 3 6 contain carried 415:421 arg2 synthesis 455:463 preparative scale GDP-l-Gal synthesis 427:463 preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase 427:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 4 7 theme A. thaliana 619:629 arg1 AtFucTA 656:662 AtFucTA 656:662 AtFucTA 656:662 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 4 7 theme A. thaliana 619:629 arg1 α1,3-fucosyltransferase 631:653 A. thaliana α1,3-fucosyltransferase 619:653 A. thaliana α1,3-fucosyltransferase (AtFucTA) 619:663 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 4 8 theme α1,6-fucosyltransferase 582:604 arg1 assay 567:571 the l-galactosylation assay 545:571 the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA) 545:663 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 1 9 theme mur1 180:183 arg1 mutant 185:190 the l-Fuc deficient mur1 mutant 160:190 the l-Fuc deficient mur1 mutant of Arabidopsis thaliana 160:214 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 6 10 theme AtFucTA 904:910 arg1 values 883:888 the apparent Km values 867:888 the apparent Km values of MmFUT8 and AtFucTA 867:910 In addition, the apparent Km values of MmFUT8 and AtFucTA suggest that l-Fuc was preferentially transferred to N-glycan compared with l-Gal by fucosyltransferases. 27993676 3 11 theme GDP-l-Gal 445:453 arg1 synthesis 455:463 preparative scale GDP-l-Gal synthesis 427:463 preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase 427:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 0 12 contain containing 38:47 arg1 N-glycans 28:36 N-glycans 28:36 N-glycans containing core l-galactose 28:64 Fucosyltransferases produce N-glycans containing core l-galactose. 27993676 0 12 contain containing 38:47 arg2 l-galactose 54:64 core l-galactose 49:64 core l-galactose 49:64 Fucosyltransferases produce N-glycans containing core l-galactose. 27993676 1 13 contain containing 87:96 arg1 l-Gal 80:84 l-Gal 80:84 l-Gal 80:84 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 13 contain containing 87:96 arg2 N-glycans 98:106 N-glycans 98:106 N-glycans 98:106 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 1 13 contain containing 87:96 arg1 l-Galactose 67:77 l-Galactose 67:77 l-Galactose (l-Gal) containing N-glycans 67:106 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 5 14 theme l-galactosylation 698:714 arg1 activity 716:723 l-galactosylation activity 698:723 l-galactosylation activity 698:723 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 6 15 theme MmFUT8 893:898 arg1 values 883:888 the apparent Km values 867:888 the apparent Km values of MmFUT8 and AtFucTA 867:910 In addition, the apparent Km values of MmFUT8 and AtFucTA suggest that l-Fuc was preferentially transferred to N-glycan compared with l-Gal by fucosyltransferases. 27993676 3 16 theme preparative 427:437 arg1 synthesis 455:463 preparative scale GDP-l-Gal synthesis 427:463 preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase 427:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 4 17 theme mouse 576:580 arg1 α1,6-fucosyltransferase 582:604 mouse α1,6-fucosyltransferase 576:604 mouse α1,6-fucosyltransferase (MmFUT8) 576:613 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 4 17 theme mouse 576:580 arg1 MmFUT8 607:612 MmFUT8 607:612 MmFUT8 607:612 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 5 18 theme N-glycan 818:825 arg1 N-acetyl-β-d-glucosamine 761:784 asparagine-linked N-acetyl-β-d-glucosamine 743:784 asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation 743:851 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 2 19 theme related 304:310 arg1 analog 312:317 a structurally related analog 289:317 a structurally related analog of GDP-l-Fuc 289:330 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 2 19 theme related 304:310 arg1 GDP-l-Gal 269:277 GDP-l-Gal 269:277 GDP-l-Gal 269:277 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 1 20 theme thaliana 207:214 arg1 mutant 185:190 the l-Fuc deficient mur1 mutant 160:190 the l-Fuc deficient mur1 mutant of Arabidopsis thaliana 160:214 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 7 21 theme N-glycan 1145:1152 arg1 in vitro 1154:1161 l-Gal containing N-glycan in vitro 1128:1161 l-Gal containing N-glycan in vitro 1128:1161 Our results clearly demonstrate that MmFUT8 and AtFucTA transfer l-Gal residues from GDP-l-Gal and synthesize l-Gal containing N-glycan in vitro. 27993676 1 22 theme wall 117:120 arg1 polysaccharides 122:136 cell wall polysaccharides 112:136 cell wall polysaccharides 112:136 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 3 23 theme A. thaliana 483:493 arg1 GDP-mannose-3 495:507 recombinant A. thaliana GDP-mannose-3' 471:508 recombinant A. thaliana GDP-mannose-3' 471:508 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 3 23 theme A. thaliana 483:493 arg1 5'-epimerase 510:521 5'-epimerase 510:521 5'-epimerase 510:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 1 24 theme Arabidopsis 195:205 arg1 thaliana 207:214 Arabidopsis thaliana 195:214 Arabidopsis thaliana 195:214 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 0 25 theme core 49:52 arg1 l-galactose 54:64 core l-galactose 49:64 core l-galactose 49:64 Fucosyltransferases produce N-glycans containing core l-galactose. 27993676 4 26 theme l-galactosylation 549:565 arg1 assay 567:571 the l-galactosylation assay 545:571 the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA) 545:663 We also demonstrated the l-galactosylation assay of mouse α1,6-fucosyltransferase (MmFUT8) and A. thaliana α1,3-fucosyltransferase (AtFucTA). 27993676 5 27 theme l-fucosylation 838:851 arg1 N-acetyl-β-d-glucosamine 761:784 asparagine-linked N-acetyl-β-d-glucosamine 743:784 asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation 743:851 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 5 28 theme asparagine-linked 743:759 arg1 N-acetyl-β-d-glucosamine 761:784 asparagine-linked N-acetyl-β-d-glucosamine 743:784 asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation 743:851 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 3 29 theme scale 439:443 arg1 synthesis 455:463 preparative scale GDP-l-Gal synthesis 427:463 preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase 427:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 6 30 theme Km 880:881 arg1 values 883:888 the apparent Km values 867:888 the apparent Km values of MmFUT8 and AtFucTA 867:910 In addition, the apparent Km values of MmFUT8 and AtFucTA suggest that l-Fuc was preferentially transferred to N-glycan compared with l-Gal by fucosyltransferases. 27993676 2 31 theme in vitrol-galactosylation 337:361 arg1 activity 363:370 in vitrol-galactosylation activity 337:370 in vitrol-galactosylation activity 337:370 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 2 32 theme l-Gal 221:225 arg1 residue 227:233 The l-Gal residue 217:233 The l-Gal residue 217:233 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 6 33 theme apparent 871:878 arg1 values 883:888 the apparent Km values 867:888 the apparent Km values of MmFUT8 and AtFucTA 867:910 In addition, the apparent Km values of MmFUT8 and AtFucTA suggest that l-Fuc was preferentially transferred to N-glycan compared with l-Gal by fucosyltransferases. 27993676 5 34 link asparagine-linked 743:759 arg1 N-acetyl-β-d-glucosamine 761:784 asparagine-linked N-acetyl-β-d-glucosamine 743:784 asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation 743:851 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 1 35 theme cell 112:115 arg1 polysaccharides 122:136 cell wall polysaccharides 112:136 cell wall polysaccharides 112:136 l-Galactose (l-Gal) containing N-glycans and cell wall polysaccharides have been detected in the l-Fuc deficient mur1 mutant of Arabidopsis thaliana. 27993676 5 36 theme asialo-agalacto-bi-antennary 789:816 arg1 N-glycan 818:825 asialo-agalacto-bi-antennary N-glycan 789:825 asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation 789:851 Both fucosyltransferases showed l-galactosylation activity from GDP-l-Gal to asparagine-linked N-acetyl-β-d-glucosamine of asialo-agalacto-bi-antennary N-glycan instead of l-fucosylation. 27993676 7 37 theme containing 1134:1143 arg1 in vitro 1154:1161 l-Gal containing N-glycan in vitro 1128:1161 l-Gal containing N-glycan in vitro 1128:1161 Our results clearly demonstrate that MmFUT8 and AtFucTA transfer l-Gal residues from GDP-l-Gal and synthesize l-Gal containing N-glycan in vitro. 27993676 2 38 theme GDP-l-Fuc 322:330 arg1 analog 312:317 a structurally related analog 289:317 a structurally related analog of GDP-l-Fuc 289:330 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 2 38 theme GDP-l-Fuc 322:330 arg1 GDP-l-Gal 269:277 GDP-l-Gal 269:277 GDP-l-Gal 269:277 The l-Gal residue is thought to be transferred from GDP-l-Gal, which is a structurally related analog of GDP-l-Fuc, but in vitrol-galactosylation activity has never been detected. 27993676 3 39 theme recombinant 471:481 arg1 GDP-mannose-3 495:507 recombinant A. thaliana GDP-mannose-3' 471:508 recombinant A. thaliana GDP-mannose-3' 471:508 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 27993676 3 39 theme recombinant 471:481 arg1 5'-epimerase 510:521 5'-epimerase 510:521 5'-epimerase 510:521 In this study, we carried out preparative scale GDP-l-Gal synthesis using recombinant A. thaliana GDP-mannose-3',5'-epimerase. 28745859 8 0 theme protein 1463:1469 arg1 folding 1471:1477 protein folding 1463:1477 protein folding 1463:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 3 1 theme proteins 477:484 arg1 folding 466:472 the folding 462:472 the folding of proteins 462:484 One gap remaining in our understanding of O-glycoproteins is how O-glycans might affect the folding of proteins. 28745859 6 2 theme step 1089:1092 arg1 kinetics 1072:1079 the kinetics 1068:1079 the kinetics of each step in unique ways 1068:1107 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 7 3 gly O-glycoproteins 1302:1316 arg1 O-glycoproteins 1302:1316 O-glycoproteins 1302:1316 O-glycoproteins 1302:1316 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 0 4 from Effects 13:19 arg1 Folding 52:58 Protein Folding 44:58 Protein Folding 44:58 Quantitative Effects of O-Linked Glycans on Protein Folding. 28745859 8 5 theme glycosylation 1409:1421 arg1 effects 1398:1404 the effects 1394:1404 the effects of glycosylation on more biological processes related to protein folding 1394:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 5 6 theme overall 854:860 arg1 rate 870:873 overall folding rate 854:873 overall folding rate 854:873 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 1 7 theme proteins 161:168 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 7 theme proteins 161:168 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 8 8 theme biological 1431:1440 arg1 processes 1442:1450 more biological processes 1426:1450 more biological processes related to protein folding 1426:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 4 9 theme binding 735:741 arg1 CBM 751:753 CBM 751:753 CBM 751:753 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 9 theme binding 735:741 arg1 module 743:748 a carbohydrate binding module 720:748 a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 720:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 9 theme binding 735:741 arg1 O-glycoprotein 704:717 a model O-glycoprotein 696:717 a model O-glycoprotein 696:717 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 10 attach derived 756:762 arg2 CBM 751:753 CBM 751:753 CBM 751:753 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 10 attach derived 756:762 arg1 TrCel7A 802:808 the Trichoderma reesei cellulase TrCel7A 769:808 the Trichoderma reesei cellulase TrCel7A 769:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 10 attach derived 756:762 arg2 module 743:748 a carbohydrate binding module 720:748 a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 720:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 10 attach derived 756:762 arg2 O-glycoprotein 704:717 a model O-glycoprotein 696:717 a model O-glycoprotein 696:717 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 11 gly O-glycoprotein 704:717 arg1 module 743:748 a carbohydrate binding module 720:748 a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 720:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 11 gly O-glycoprotein 704:717 arg1 O-glycoprotein 704:717 a model O-glycoprotein 696:717 a model O-glycoprotein 696:717 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 1 12 theme Protein 61:67 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 12 theme Protein 61:67 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 5 13 from increase 842:849 arg1 rate 870:873 overall folding rate 854:873 overall folding rate 854:873 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 2 14 theme proteins 340:347 arg1 properties 322:331 physical properties 313:331 physical properties 313:331 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 2 14 theme proteins 340:347 arg1 function 299:306 function 299:306 function 299:306 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 2 14 theme proteins 340:347 arg1 structure 288:296 structure 288:296 structure 288:296 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 6 15 theme glycosylation 1036:1048 arg1 patterns 1050:1057 glycosylation patterns 1036:1057 glycosylation patterns 1036:1057 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 8 16 from effects 1398:1404 arg1 processes 1442:1450 more biological processes 1426:1450 more biological processes related to protein folding 1426:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 8 17 theme related 1452:1458 arg1 processes 1442:1450 more biological processes 1426:1450 more biological processes related to protein folding 1426:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 6 18 theme unique 1097:1102 arg1 ways 1104:1107 unique ways 1097:1107 unique ways 1097:1107 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 5 19 gly glycosylation 937:949 arg2 site 951:954 the glycosylation site 933:954 the glycosylation site 933:954 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 5 20 theme folding 862:868 arg1 rate 870:873 overall folding rate 854:873 overall folding rate 854:873 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 4 21 theme glycosylation 574:586 arg1 patterns 588:595 certain glycosylation patterns 566:595 certain glycosylation patterns 566:595 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 3 22 theme O-glycoproteins 416:430 arg1 understanding 399:411 our understanding 395:411 our understanding of O-glycoproteins 395:430 One gap remaining in our understanding of O-glycoproteins is how O-glycans might affect the folding of proteins. 28745859 4 23 dep synthetic 514:522 arg1 homogeneous 525:535 homogeneous 525:535 homogeneous 525:535 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 24 theme O-glycoprotein 704:717 arg1 pathway 685:691 the folding pathway 673:691 the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 673:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 6 25 from ways 1104:1107 arg1 kinetics 1072:1079 the kinetics 1068:1079 the kinetics of each step in unique ways 1068:1107 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 4 26 theme folding 651:657 arg1 machinery 659:667 any cellular folding machinery 638:667 any cellular folding machinery 638:667 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 0 27 theme Quantitative 0:11 arg1 Effects 13:19 Quantitative Effects 0:19 Quantitative Effects of O-Linked Glycans on Protein Folding 0:58 Quantitative Effects of O-Linked Glycans on Protein Folding. 28745859 1 28 theme diverse 90:96 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 28 theme diverse 90:96 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 4 29 theme cellular 642:649 arg1 machinery 659:667 any cellular folding machinery 638:667 any cellular folding machinery 638:667 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 0 30 theme O-Linked 24:31 arg1 Glycans 33:39 O-Linked Glycans 24:39 O-Linked Glycans 24:39 Quantitative Effects of O-Linked Glycans on Protein Folding. 28745859 4 31 theme synthetic 514:522 arg1 O-glycopeptides 537:551 synthetic, homogeneous O-glycopeptides 514:551 synthetic, homogeneous O-glycopeptides 514:551 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 5 32 theme single 896:901 arg1 O-mannose 903:911 a single O-mannose 894:911 a single O-mannose 894:911 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 5 32 theme single 896:901 arg1 glycan 921:926 glycan 921:926 glycan 921:926 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 4 33 theme Trichoderma 773:783 arg1 cellulase 792:800 the Trichoderma reesei cellulase 769:800 the Trichoderma reesei cellulase TrCel7A 769:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 1 34 theme those 190:194 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 34 theme those 190:194 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 4 35 theme model 698:702 arg1 module 743:748 a carbohydrate binding module 720:748 a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 720:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 35 theme model 698:702 arg1 O-glycoprotein 704:717 a model O-glycoprotein 696:717 a model O-glycoprotein 696:717 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 36 theme O-glycopeptides 537:551 arg1 advantage 501:509 advantage 501:509 advantage of synthetic, homogeneous O-glycopeptides 501:551 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 6 37 from kinetics 1072:1079 arg1 ways 1104:1107 unique ways 1097:1107 unique ways 1097:1107 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 8 38 theme future 1367:1372 arg1 investigations 1374:1387 future investigations 1367:1387 future investigations into the effects of glycosylation on more biological processes related to protein folding 1367:1477 Such an understanding is expected to facilitate future investigations into the effects of glycosylation on more biological processes related to protein folding. 28745859 1 39 theme common 99:104 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 39 theme common 99:104 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 6 40 from step 1089:1092 arg1 ways 1104:1107 unique ways 1097:1107 unique ways 1097:1107 We were also able to show that glycosylation patterns affected the kinetics of each step in unique ways, which may help to explain the observations made here. 28745859 4 41 theme intrinsic 605:613 arg1 effect 615:620 an intrinsic effect 602:620 an intrinsic effect 602:620 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 7 42 theme quantitative 1197:1208 arg1 understanding 1210:1222 quantitative understanding 1197:1222 quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins 1197:1316 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 4 43 theme folding 677:683 arg1 pathway 685:691 the folding pathway 673:691 the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 673:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 2 44 theme Much 233:236 arg1 work 238:241 Much work 233:241 Much work 233:241 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 5 45 theme 6-fold 835:840 arg1 increase 842:849 a 6-fold increase 833:849 a 6-fold increase in overall folding rate 833:873 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 5 45 theme 6-fold 835:840 arg1 effect 825:830 The strongest effect 811:830 The strongest effect 811:830 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 4 46 theme independent 623:633 arg1 effect 615:620 an intrinsic effect 602:620 an intrinsic effect 602:620 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 0 47 theme Glycans 33:39 arg1 Effects 13:19 Quantitative Effects 0:19 Quantitative Effects of O-Linked Glycans on Protein Folding 0:58 Quantitative Effects of O-Linked Glycans on Protein Folding. 28745859 4 48 theme carbohydrate 722:733 arg1 CBM 751:753 CBM 751:753 CBM 751:753 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 48 theme carbohydrate 722:733 arg1 module 743:748 a carbohydrate binding module 720:748 a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A 720:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 48 theme carbohydrate 722:733 arg1 O-glycoprotein 704:717 a model O-glycoprotein 696:717 a model O-glycoprotein 696:717 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 5 49 theme strongest 815:823 arg1 increase 842:849 a 6-fold increase 833:849 a 6-fold increase in overall folding rate 833:873 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 5 49 theme strongest 815:823 arg1 effect 825:830 The strongest effect 811:830 The strongest effect 811:830 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 4 50 theme cellulase 792:800 arg1 TrCel7A 802:808 the Trichoderma reesei cellulase TrCel7A 769:808 the Trichoderma reesei cellulase TrCel7A 769:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 7 51 theme first 1179:1183 arg1 work 1169:1172 This work 1164:1172 This work 1164:1172 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 7 51 theme first 1179:1183 arg1 step 1185:1188 a first step 1177:1188 a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins 1177:1316 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 0 52 theme Protein 44:50 arg1 Folding 52:58 Protein Folding 44:58 Protein Folding 44:58 Quantitative Effects of O-Linked Glycans on Protein Folding. 28745859 2 53 attach attached 364:371 arg2 it 358:359 it 358:359 it 358:359 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 2 53 attach attached 364:371 arg1 proteins 340:347 the proteins 336:347 the proteins to which it is attached 336:371 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 4 54 theme reesei 785:790 arg1 cellulase 792:800 the Trichoderma reesei cellulase 769:800 the Trichoderma reesei cellulase TrCel7A 769:808 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 2 55 dep structure 288:296 arg1 the 284:286 the 284:286 the 284:286 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 3 56 gly O-glycoproteins 416:430 arg1 O-glycoproteins 416:430 O-glycoproteins 416:430 O-glycoproteins 416:430 One gap remaining in our understanding of O-glycoproteins is how O-glycans might affect the folding of proteins. 28745859 4 57 contain have 597:600 arg1 patterns 588:595 certain glycosylation patterns 566:595 certain glycosylation patterns 566:595 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 57 contain have 597:600 arg2 effect 615:620 an intrinsic effect 602:620 an intrinsic effect 602:620 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 58 gly O-glycopeptides 537:551 arg2 O-glycopeptides 537:551 synthetic, homogeneous O-glycopeptides 514:551 synthetic, homogeneous O-glycopeptides 514:551 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 1 59 theme important 111:119 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 59 theme important 111:119 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 7 60 theme O-glycoproteins 1302:1316 arg1 folding 1291:1297 the folding 1287:1297 the folding of O-glycoproteins 1287:1316 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 5 61 theme peptide 987:993 arg1 sequence 995:1002 the peptide sequence 983:1002 the peptide sequence 983:1002 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 7 62 theme intrinsic 1270:1278 arg1 means 1280:1284 intrinsic means 1270:1284 intrinsic means 1270:1284 This work is a first step toward quantitative understanding of how O-glycosylation might control, through intrinsic means, the folding of O-glycoproteins. 28745859 1 63 theme post-translational 121:138 arg1 O-glycosylation 69:83 Protein O-glycosylation 61:83 Protein O-glycosylation 61:83 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 1 63 theme post-translational 121:138 arg1 modification 140:151 a diverse, common, and important post-translational modification 88:151 a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound 88:230 Protein O-glycosylation is a diverse, common, and important post-translational modification of both proteins inside the cell and those that are secreted or membrane-bound. 28745859 2 64 theme physical 313:320 arg1 properties 322:331 physical properties 313:331 physical properties 313:331 Much work has shown that O-glycosylation can alter the structure, function, and physical properties of the proteins to which it is attached. 28745859 5 65 theme sequence 995:1002 arg1 N-terminus 969:978 the N-terminus 965:978 the N-terminus of the peptide sequence 965:1002 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 5 66 theme glycosylation 937:949 arg1 site 951:954 the glycosylation site 933:954 the glycosylation site 933:954 The strongest effect, a 6-fold increase in overall folding rate, was observed when a single O-mannose was the glycan, and the glycosylation site was near the N-terminus of the peptide sequence. 28745859 4 67 theme certain 566:572 arg1 patterns 588:595 certain glycosylation patterns 566:595 certain glycosylation patterns 566:595 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 28745859 4 68 theme machinery 659:667 arg1 independent 623:633 independent 623:633 independent 623:633 Here, we took advantage of synthetic, homogeneous O-glycopeptides to show that certain glycosylation patterns have an intrinsic effect, independent of any cellular folding machinery, on the folding pathway of a model O-glycoprotein, a carbohydrate binding module (CBM) derived from the Trichoderma reesei cellulase TrCel7A. 26603318 5 0 theme thermal 1126:1132 arg1 unfolding 1178:1186 thermal and guanidine hydrochloride (GdnCl)-induced unfolding 1126:1186 unfolding 1178:1186 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 4 1 theme pH 1068:1069 arg1 range 1071:1075 the acidic pH range 1057:1075 the acidic pH range for the deglycosylated and the N110Q protein 1057:1120 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 3 2 theme deglycosylated 465:478 arg1 lectin 507:512 a deglycosylated Escherichia coli expressed lectin 463:512 a deglycosylated Escherichia coli expressed lectin 463:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 4 3 theme sugar-binding 869:881 arg1 properties 883:892 sugar-binding properties 869:892 sugar-binding properties 869:892 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 3 4 theme high-mannose 515:526 arg1 characteristics 444:458 the characteristics 440:458 the characteristics of a deglycosylated Escherichia coli expressed lectin 440:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 4 theme high-mannose 515:526 arg1 oligosaccharides 528:543 high-mannose oligosaccharides 515:543 high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 515:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 5 5 theme higher 1236:1241 arg1 stability 1243:1251 higher stability 1236:1251 higher stability 1236:1251 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 7 6 theme active 1823:1828 arg1 conformation 1830:1841 the active conformation 1819:1841 the active conformation 1819:1841 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 3 7 theme lectin 507:512 arg1 characteristics 444:458 the characteristics 440:458 the characteristics of a deglycosylated Escherichia coli expressed lectin 440:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 7 theme lectin 507:512 arg1 oligosaccharides 528:543 high-mannose oligosaccharides 515:543 high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 515:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 8 dep mutants 587:593 arg1 mutants 587:593 its glycosylation mutants 569:593 its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 569:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 8 dep mutants 587:593 arg1 N110Q 604:608 N110Q 604:608 N110Q 604:608 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 8 dep mutants 587:593 arg1 N66Q 595:598 N66Q 595:598 N66Q 595:598 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 4 9 theme recombinant 931:941 arg1 lectins 943:949 wild-type and aforementioned recombinant lectins 902:949 wild-type and aforementioned recombinant lectins 902:949 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 6 10 gly deglycosylated 1478:1491 arg1 proteins 1493:1500 deglycosylated proteins 1478:1500 deglycosylated proteins 1478:1500 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 2 11 theme glycans 285:291 arg1 role 264:267 the role 260:267 the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability 260:407 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 6 12 theme wild 1556:1559 arg1 type 1561:1564 the wild type 1552:1564 the wild type 1552:1564 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 2 13 attach attached 293:300 arg1 Asn 329:331 Asn 110 329:335 Asn 110 329:335 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 2 13 attach attached 293:300 arg2 glycans 285:291 complex-type glycans 272:291 complex-type glycans attached to asparagine (Asn) 66 and Asn 110 272:335 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 2 13 attach attached 293:300 arg1 Asn 317:319 asparagine (Asn) 66 305:323 asparagine (Asn) 66 305:323 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 3 14 dep Escherichia 480:490 arg1 coli 492:495 coli 492:495 coli 492:495 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 15 from asparagines 553:563 arg1 characteristics 444:458 the characteristics 440:458 the characteristics of a deglycosylated Escherichia coli expressed lectin 440:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 15 from asparagines 553:563 arg1 oligosaccharides 528:543 high-mannose oligosaccharides 515:543 high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 515:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 1 16 theme non-seed 134:141 arg1 portions 143:150 non-seed portions 134:150 non-seed portions of turmeric 134:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 5 17 theme recombinant 1286:1296 arg1 lectin 1298:1303 the deglycosylated recombinant lectin 1267:1303 the deglycosylated recombinant lectin 1267:1303 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 0 18 theme glycosylation 60:72 arg1 Role 43:46 lectin: Role 35:46 lectin: Role of N-linked glycosylation 35:72 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 5 19 theme hydrochloride 1148:1160 arg1 unfolding 1178:1186 thermal and guanidine hydrochloride (GdnCl)-induced unfolding 1126:1186 unfolding 1178:1186 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 7 20 from N-glycosylation 1659:1673 arg1 lectin 1687:1692 turmeric lectin 1678:1692 turmeric lectin 1678:1692 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 7 21 theme proper 1734:1739 arg1 folding 1741:1747 its proper folding 1730:1747 its proper folding upon changes in pH 1730:1766 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 4 22 theme reduced 979:985 arg1 activity 1001:1008 reduced agglutination activity 979:1008 reduced agglutination activity 979:1008 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 3 23 gly deglycosylated 465:478 arg1 lectin 507:512 a deglycosylated Escherichia coli expressed lectin 463:512 a deglycosylated Escherichia coli expressed lectin 463:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 6 24 theme thermal 1445:1451 arg1 denaturation 1453:1464 thermal denaturation 1445:1464 thermal denaturation 1445:1464 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 7 25 from aggregation 1855:1865 arg1 molecules 1899:1907 unfolded or partially folded molecules 1870:1907 unfolded or partially folded molecules 1870:1907 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 4 26 theme circular 728:735 arg1 dichroism 737:745 UV circular dichroism 725:745 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 26 theme circular 728:735 arg1 CD 748:749 CD 748:749 CD 748:749 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 1 27 theme longa 83:87 arg1 protein 123:129 a mannose-binding protein 105:129 a mannose-binding protein of non-seed portions of turmeric 105:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 27 theme longa 83:87 arg1 lectin 97:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 28 theme acidic 1061:1066 arg1 range 1071:1075 the acidic pH range 1057:1075 the acidic pH range for the deglycosylated and the N110Q protein 1057:1120 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 3 29 theme native 425:430 arg1 lectin 432:437 the native lectin 421:437 the native lectin 421:437 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 0 30 theme Curcuma 13:19 arg1 longa 21:25 Curcuma longa 13:25 Curcuma longa 13:25 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 5 31 theme wild-type 1193:1201 arg1 lectins 1220:1226 the wild-type and high-mannose lectins 1189:1226 the wild-type and high-mannose lectins 1189:1226 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 3 32 from mutants 587:593 arg1 characteristics 444:458 the characteristics 440:458 the characteristics of a deglycosylated Escherichia coli expressed lectin 440:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 32 from mutants 587:593 arg1 oligosaccharides 528:543 high-mannose oligosaccharides 515:543 high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 515:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 4 33 theme tertiary 1022:1029 arg1 structure 1031:1039 tertiary structure 1022:1039 tertiary structure 1022:1039 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 5 34 theme higher 1340:1345 arg1 Tm 1347:1348 a higher Tm 1338:1348 a higher Tm of denaturation 1338:1364 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 5 35 theme GdnCl 1163:1167 arg1 unfolding 1178:1186 thermal and guanidine hydrochloride (GdnCl)-induced unfolding 1126:1186 unfolding 1178:1186 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 7 36 gly N-glycosylation 1659:1673 arg1 lectin 1687:1692 turmeric lectin 1678:1692 turmeric lectin 1678:1692 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 7 36 gly N-glycosylation 1659:1673 arg1 folding 1741:1747 its proper folding 1730:1747 its proper folding upon changes in pH 1730:1766 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 3 37 theme Pichia 623:628 arg1 pastoris 630:637 Pichia pastoris 623:637 Pichia pastoris 623:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 6 38 theme Reversibility 1413:1425 arg1 experiments 1427:1437 Reversibility experiments 1413:1437 Reversibility experiments after thermal denaturation 1413:1464 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 5 39 theme -induced 1169:1176 arg1 unfolding 1178:1186 thermal and guanidine hydrochloride (GdnCl)-induced unfolding 1126:1186 unfolding 1178:1186 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 0 40 link N-linked 51:58 arg1 glycosylation 60:72 N-linked glycosylation 51:72 N-linked glycosylation 51:72 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 5 41 gly deglycosylated 1271:1284 arg1 lectin 1298:1303 the deglycosylated recombinant lectin 1267:1303 the deglycosylated recombinant lectin 1267:1303 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 4 42 theme emission 774:781 arg1 maximum 783:789 fluorescence emission maximum 761:789 fluorescence emission maximum 761:789 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 42 theme emission 774:781 arg1 spectra 752:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 1 43 theme antibacterial 194:206 arg1 activities 237:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 7 44 theme folded 1892:1897 arg1 molecules 1899:1907 unfolded or partially folded molecules 1870:1907 unfolded or partially folded molecules 1870:1907 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 4 45 theme N110Q 1108:1112 arg1 protein 1114:1120 the N110Q protein 1104:1120 the N110Q protein 1104:1120 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 5 46 theme greater 1371:1377 arg1 change 1391:1396 a greater free energy change 1369:1396 a greater free energy change 1369:1396 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 3 47 theme Escherichia 480:490 arg1 lectin 507:512 a deglycosylated Escherichia coli expressed lectin 463:512 a deglycosylated Escherichia coli expressed lectin 463:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 1 48 theme α-glucosidase 212:224 arg1 activities 237:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 5 49 theme energy 1384:1389 arg1 change 1391:1396 a greater free energy change 1369:1396 a greater free energy change 1369:1396 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 5 50 theme high-mannose 1207:1218 arg1 lectins 1220:1226 the wild-type and high-mannose lectins 1189:1226 the wild-type and high-mannose lectins 1189:1226 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 4 51 theme tertiary 846:853 arg1 structures 855:864 secondary or tertiary structures 833:864 secondary or tertiary structures 833:864 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 2 52 theme complex-type 272:283 arg1 glycans 285:291 complex-type glycans 272:291 complex-type glycans attached to asparagine (Asn) 66 and Asn 110 272:335 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 7 53 from changes 1754:1760 arg1 pH 1765:1766 pH 1765:1766 pH 1765:1766 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 3 54 theme expressed 497:505 arg1 lectin 507:512 a deglycosylated Escherichia coli expressed lectin 463:512 a deglycosylated Escherichia coli expressed lectin 463:512 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 6 55 theme greater 1579:1585 arg1 recovery 1587:1594 a much greater recovery 1572:1594 a much greater recovery to the native state upon refolding 1572:1629 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 1 56 theme turmeric 155:162 arg1 portions 143:150 non-seed portions 134:150 non-seed portions of turmeric 134:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 5 57 contain possess 1228:1234 arg1 lectins 1220:1226 the wild-type and high-mannose lectins 1189:1226 the wild-type and high-mannose lectins 1189:1226 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 5 57 contain possess 1228:1234 arg2 stability 1243:1251 higher stability 1236:1251 higher stability 1236:1251 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 2 58 theme asparagine 305:314 arg1 Asn 317:319 asparagine (Asn) 66 305:323 asparagine (Asn) 66 305:323 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 4 59 theme secondary 833:841 arg1 structures 855:864 secondary or tertiary structures 833:864 secondary or tertiary structures 833:864 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 60 theme aforementioned 916:929 arg1 lectins 943:949 wild-type and aforementioned recombinant lectins 902:949 wild-type and aforementioned recombinant lectins 902:949 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 3 61 theme glycosylation 573:585 arg1 mutants 587:593 its glycosylation mutants 569:593 its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris 569:637 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 61 theme glycosylation 573:585 arg1 N110Q 604:608 N110Q 604:608 N110Q 604:608 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 3 61 theme glycosylation 573:585 arg1 N66Q 595:598 N66Q 595:598 N66Q 595:598 Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. 26603318 1 62 theme antifungal 182:191 arg1 activities 237:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 63 theme mannose-binding 107:121 arg1 protein 123:129 a mannose-binding protein 105:129 a mannose-binding protein of non-seed portions of turmeric 105:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 63 theme mannose-binding 107:121 arg1 lectin 97:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 64 theme wild-type 902:910 arg1 lectins 943:949 wild-type and aforementioned recombinant lectins 902:949 wild-type and aforementioned recombinant lectins 902:949 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 0 65 theme N-linked 51:58 arg1 glycosylation 60:72 N-linked glycosylation 51:72 N-linked glycosylation 51:72 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 6 66 theme native 1603:1608 arg1 state 1610:1614 the native state 1599:1614 the native state 1599:1614 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 4 67 theme agglutination 987:999 arg1 activity 1001:1008 reduced agglutination activity 979:1008 reduced agglutination activity 979:1008 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 5 68 theme deglycosylated 1271:1284 arg1 lectin 1298:1303 the deglycosylated recombinant lectin 1267:1303 the deglycosylated recombinant lectin 1267:1303 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 5 69 theme guanidine 1138:1146 arg1 unfolding 1178:1186 thermal and guanidine hydrochloride (GdnCl)-induced unfolding 1126:1186 unfolding 1178:1186 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 1 70 theme portions 143:150 arg1 protein 123:129 a mannose-binding protein 105:129 a mannose-binding protein of non-seed portions of turmeric 105:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 70 theme portions 143:150 arg1 lectin 97:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 71 located observed 1045:1052 arg2 loss 1014:1017 loss 1014:1017 loss of tertiary structure 1014:1039 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 71 located observed 1045:1052 arg2 activity 1001:1008 reduced agglutination activity 979:1008 reduced agglutination activity 979:1008 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 71 located observed 1045:1052 arg1 range 1071:1075 the acidic pH range 1057:1075 the acidic pH range for the deglycosylated and the N110Q protein 1057:1120 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 7 72 theme folding 1741:1747 arg1 maintenance 1715:1725 the maintenance 1711:1725 the maintenance of its proper folding upon changes in pH 1711:1766 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 5 73 theme denaturation 1353:1364 arg1 change 1391:1396 a greater free energy change 1369:1396 a greater free energy change 1369:1396 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 5 73 theme denaturation 1353:1364 arg1 Tm 1347:1348 a higher Tm 1338:1348 a higher Tm of denaturation 1338:1364 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 6 74 theme deglycosylated 1478:1491 arg1 proteins 1493:1500 deglycosylated proteins 1478:1500 deglycosylated proteins 1478:1500 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 4 75 from change 823:828 arg1 structures 855:864 secondary or tertiary structures 833:864 secondary or tertiary structures 833:864 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 75 from change 823:828 arg1 properties 883:892 sugar-binding properties 869:892 sugar-binding properties 869:892 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 1 76 theme Curcuma 75:81 arg1 longa 83:87 Curcuma longa 75:87 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 77 theme UV 725:726 arg1 dichroism 737:745 UV circular dichroism 725:745 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 77 theme UV 725:726 arg1 CD 748:749 CD 748:749 CD 748:749 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 0 78 theme longa 21:25 arg1 Stability 0:8 Stability 0:8 Stability of Curcuma longa 0:25 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 7 79 theme turmeric 1678:1685 arg1 lectin 1687:1692 turmeric lectin 1678:1692 turmeric lectin 1678:1692 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 1 80 theme rhizome 89:95 arg1 protein 123:129 a mannose-binding protein 105:129 a mannose-binding protein of non-seed portions of turmeric 105:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 80 theme rhizome 89:95 arg1 lectin 97:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 81 theme dichroism 737:745 arg1 maximum 783:789 fluorescence emission maximum 761:789 fluorescence emission maximum 761:789 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 81 theme dichroism 737:745 arg1 assay 809:813 hemagglutination assay 792:813 hemagglutination assay 792:813 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 81 theme dichroism 737:745 arg1 spectra 752:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 2 82 theme lectin 379:384 arg1 activity 386:393 lectin activity 379:393 lectin activity 379:393 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 0 83 theme lectin 35:40 arg1 Role 43:46 lectin: Role 35:46 lectin: Role of N-linked glycosylation 35:72 Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation. 26603318 4 84 theme hemagglutination 792:807 arg1 assay 809:813 hemagglutination assay 792:813 hemagglutination assay 792:813 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 84 theme hemagglutination 792:807 arg1 spectra 752:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 2 85 theme carbohydrates 362:374 arg1 role 354:357 the role 350:357 the role of carbohydrates in lectin activity and stability 350:407 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 4 86 theme structure 1031:1039 arg1 activity 1001:1008 reduced agglutination activity 979:1008 reduced agglutination activity 979:1008 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 86 theme structure 1031:1039 arg1 loss 1014:1017 loss 1014:1017 loss of tertiary structure 1014:1039 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 2 87 from role 354:357 arg1 stability 399:407 stability 399:407 stability 399:407 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 2 87 from role 354:357 arg1 activity 386:393 lectin activity 379:393 lectin activity 379:393 We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. 26603318 1 88 contain have 177:180 arg1 protein 123:129 a mannose-binding protein 105:129 a mannose-binding protein of non-seed portions of turmeric 105:162 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 88 contain have 177:180 arg2 activities 237:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 1 88 contain have 177:180 arg1 lectin 97:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin 75:102 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 26603318 4 89 theme fluorescence 761:772 arg1 maximum 783:789 fluorescence emission maximum 761:789 fluorescence emission maximum 761:789 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 4 89 theme fluorescence 761:772 arg1 spectra 752:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra 721:758 Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. 26603318 6 90 theme thermal 1527:1533 arg1 inactivation 1535:1546 thermal inactivation 1527:1546 thermal inactivation 1527:1546 Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. 26603318 7 91 theme unfolded 1870:1877 arg1 molecules 1899:1907 unfolded or partially folded molecules 1870:1907 unfolded or partially folded molecules 1870:1907 These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. 26603318 5 92 theme free 1379:1382 arg1 change 1391:1396 a greater free energy change 1369:1396 a greater free energy change 1369:1396 In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. 26603318 1 93 theme inhibitory 226:235 arg1 activities 237:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 antifungal, antibacterial and α-glucosidase inhibitory activities 182:246 Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. 25748215 0 0 theme Bank 78:81 arg1 Effects 0:6 Effects 0:6 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 0:131 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 0 theme Bank 78:81 arg1 analysis 83:90 Protein Data Bank analysis 65:90 Protein Data Bank analysis 65:90 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 6 1 dep suggest 1100:1106 arg1 only 1112:1115 only 1112:1115 only 1112:1115 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 0 2 theme Data 73:76 arg1 Bank 78:81 Protein Data Bank 65:81 Protein Data Bank analysis 65:90 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 4 3 theme computational 739:751 arg1 approach 753:760 an integrated computational approach 725:760 an integrated computational approach of the Protein Data Bank 725:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 5 4 theme protein 1058:1064 arg1 stability 1066:1074 protein stability 1058:1074 protein stability 1058:1074 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 0 5 from Effects 0:6 arg1 conformation 38:49 protein conformation 30:49 protein conformation 30:49 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 5 from Effects 0:6 arg1 dynamics 55:62 dynamics 55:62 dynamics 55:62 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 4 6 theme molecular 820:828 arg1 simulations 839:849 atomistic molecular dynamics simulations 810:849 atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins 810:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 5 7 from increase 1046:1053 arg1 stability 1066:1074 protein stability 1058:1074 protein stability 1058:1074 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 4 8 theme atomistic 810:818 arg1 simulations 839:849 atomistic molecular dynamics simulations 810:849 atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins 810:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 9 theme Protein 769:775 arg1 Bank 782:785 the Protein Data Bank 765:785 the Protein Data Bank 765:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 0 10 theme dynamics 106:113 arg1 simulation 115:124 molecular dynamics simulation 96:124 molecular dynamics simulation study 96:130 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 3 11 from interest 395:402 arg1 developments 407:418 developments 407:418 developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties 407:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 6 12 theme common 1119:1124 arg1 role 1126:1129 a common role 1117:1129 a common role of glycosylation in proteins 1117:1158 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 5 13 theme protein 1007:1013 arg1 dynamics 1015:1022 protein dynamics 1007:1022 protein dynamics 1007:1022 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 3 14 theme structural 457:466 arg1 consequences 468:479 the structural consequences 453:479 the structural consequences of site-specific glycosylation 453:510 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 0 15 theme molecular 96:104 arg1 simulation 115:124 molecular dynamics simulation 96:124 molecular dynamics simulation study 96:130 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 3 16 theme protein 568:574 arg1 design 576:581 protein design 568:581 protein design with advantageous properties 568:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 4 17 from impacts 632:638 arg1 dynamics 678:685 dynamics 678:685 dynamics 678:685 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 17 from impacts 632:638 arg1 structure 664:672 protein structure 656:672 protein structure 656:672 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 3 18 dep approaches 431:440 arg1 predict 445:451 predict 445:451 to predict the structural consequences of site-specific glycosylation 442:510 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 3 18 dep approaches 431:440 arg1 understand 519:528 understand 519:528 to understand how these effects can be exploited in protein design with advantageous properties 516:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 4 19 theme structure 787:795 arg1 analysis 797:804 structure analysis 787:804 structure analysis 787:804 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 6 20 theme glycosylation 1134:1146 arg1 role 1126:1129 a common role 1117:1129 a common role of glycosylation in proteins 1117:1158 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 6 20 theme glycosylation 1134:1146 arg1 need 1172:1175 a need 1170:1175 a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties 1170:1267 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 1 21 theme important 175:183 arg1 modifications 240:252 the most important, chemically complex, and ubiquitous post-translational modifications 166:252 the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes 166:270 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 6 22 gly glycosylated 1213:1224 arg1 need 1172:1175 a need 1170:1175 a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties 1170:1267 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 6 23 from role 1126:1129 arg1 proteins 1151:1158 proteins 1151:1158 proteins 1151:1158 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 4 24 theme Bank 782:785 arg1 approach 753:760 an integrated computational approach 725:760 an integrated computational approach of the Protein Data Bank 725:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 24 theme Bank 782:785 arg1 simulations 839:849 atomistic molecular dynamics simulations 810:849 atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins 810:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 3 25 theme considerable 382:393 arg1 interest 395:402 considerable interest 382:402 considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties 382:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 3 26 theme advantageous 588:599 arg1 properties 601:610 advantageous properties 588:610 advantageous properties 588:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 3 27 theme glycosylation 498:510 arg1 consequences 468:479 the structural consequences 453:479 the structural consequences of site-specific glycosylation 453:510 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 0 28 theme N-glycosylation 11:25 arg1 Effects 0:6 Effects 0:6 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 0:131 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 28 theme N-glycosylation 11:25 arg1 study 126:130 molecular dynamics simulation study 96:130 molecular dynamics simulation study 96:130 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 28 theme N-glycosylation 11:25 arg1 analysis 83:90 Protein Data Bank analysis 65:90 Protein Data Bank analysis 65:90 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 2 29 attach linked 307:312 arg2 N-glycans 277:285 The N-glycans 273:285 The N-glycans that are covalently linked to proteins 273:324 The N-glycans that are covalently linked to proteins are involved in numerous biological processes. 25748215 2 29 attach linked 307:312 arg1 proteins 317:324 proteins 317:324 proteins 317:324 The N-glycans that are covalently linked to proteins are involved in numerous biological processes. 25748215 0 30 theme simulation 115:124 arg1 Effects 0:6 Effects 0:6 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 0:131 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 30 theme simulation 115:124 arg1 study 126:130 molecular dynamics simulation study 96:130 molecular dynamics simulation study 96:130 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 6 31 theme intrinsic 1240:1248 arg1 properties 1258:1267 their intrinsic dynamic properties 1234:1267 their intrinsic dynamic properties 1234:1267 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 4 32 theme protein 656:662 arg1 structure 664:672 protein structure 656:672 protein structure 656:672 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 3 33 theme approaches 431:440 arg1 developments 407:418 developments 407:418 developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties 407:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 6 34 theme certain 1181:1187 arg1 proteins 1189:1196 certain proteins 1181:1196 certain proteins 1181:1196 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 1 35 dep important 175:183 arg1 complex 197:203 complex 197:203 complex 197:203 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 0 36 theme protein 30:36 arg1 conformation 38:49 protein conformation 30:49 protein conformation 30:49 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 4 37 gly glycosylated 854:865 arg1 proteins 886:893 glycosylated and deglycosylated proteins 854:893 glycosylated and deglycosylated proteins 854:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 38 theme proteins 886:893 arg1 approach 753:760 an integrated computational approach 725:760 an integrated computational approach of the Protein Data Bank 725:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 38 theme proteins 886:893 arg1 simulations 839:849 atomistic molecular dynamics simulations 810:849 atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins 810:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 3 39 theme site-specific 484:496 arg1 glycosylation 498:510 site-specific glycosylation 484:510 site-specific glycosylation 484:510 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 3 40 with design 576:581 arg1 properties 601:610 advantageous properties 588:610 advantageous properties 588:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 5 41 theme significant 951:961 arg1 changes 963:969 significant changes 951:969 significant changes in protein structure 951:990 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 1 42 from modifications 240:252 arg1 eukaryotes 261:270 all eukaryotes 257:270 all eukaryotes 257:270 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 0 43 gly N-glycosylation 11:25 arg1 protein 30:36 protein conformation 30:49 protein conformation 30:49 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 6 44 from need 1172:1175 arg1 proteins 1151:1158 proteins 1151:1158 proteins 1151:1158 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 2 45 theme biological 351:360 arg1 processes 362:370 numerous biological processes 342:370 numerous biological processes 342:370 The N-glycans that are covalently linked to proteins are involved in numerous biological processes. 25748215 3 46 theme general 423:429 arg1 approaches 431:440 general approaches 423:440 general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties 423:610 There is considerable interest in developments of general approaches to predict the structural consequences of site-specific glycosylation and to understand how these effects can be exploited in protein design with advantageous properties. 25748215 1 47 link N-linked 133:140 arg1 glycosylation 142:154 N-linked glycosylation 133:154 N-linked glycosylation 133:154 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 2 48 theme numerous 342:349 arg1 processes 362:370 numerous biological processes 342:370 numerous biological processes 342:370 The N-glycans that are covalently linked to proteins are involved in numerous biological processes. 25748215 6 49 theme dynamic 1250:1256 arg1 properties 1258:1267 their intrinsic dynamic properties 1234:1267 their intrinsic dynamic properties 1234:1267 Overall, these results suggest not only a common role of glycosylation in proteins, but also a need for certain proteins to be properly glycosylated to gain their intrinsic dynamic properties. 25748215 4 50 theme glycosylated 854:865 arg1 proteins 886:893 glycosylated and deglycosylated proteins 854:893 glycosylated and deglycosylated proteins 854:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 51 gly deglycosylated 871:884 arg1 proteins 886:893 glycosylated and deglycosylated proteins 854:893 glycosylated and deglycosylated proteins 854:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 52 theme Data 777:780 arg1 Bank 782:785 the Protein Data Bank 765:785 the Protein Data Bank 765:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 1 53 theme ubiquitous 210:219 arg1 modifications 240:252 the most important, chemically complex, and ubiquitous post-translational modifications 166:252 the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes 166:270 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 4 54 theme dynamics 830:837 arg1 simulations 839:849 atomistic molecular dynamics simulations 810:849 atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins 810:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 4 55 dep approach 753:760 arg1 analysis 797:804 structure analysis 787:804 structure analysis 787:804 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 1 56 theme N-linked 133:140 arg1 glycosylation 142:154 N-linked glycosylation 133:154 N-linked glycosylation 133:154 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 1 57 theme post-translational 221:238 arg1 modifications 240:252 the most important, chemically complex, and ubiquitous post-translational modifications 166:252 the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes 166:270 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 0 58 theme Protein 65:71 arg1 Bank 78:81 Protein Data Bank 65:81 Protein Data Bank analysis 65:90 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 4 59 theme deglycosylated 871:884 arg1 proteins 886:893 glycosylated and deglycosylated proteins 854:893 glycosylated and deglycosylated proteins 854:893 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 5 60 from changes 963:969 arg1 structure 982:990 protein structure 974:990 protein structure 974:990 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 4 61 theme integrated 728:737 arg1 approach 753:760 an integrated computational approach 725:760 an integrated computational approach of the Protein Data Bank 725:785 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 25748215 1 62 theme modifications 240:252 arg1 modifications 240:252 the most important, chemically complex, and ubiquitous post-translational modifications 166:252 the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes 166:270 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 1 62 theme modifications 240:252 arg1 one 159:161 one 159:161 one 159:161 N-linked glycosylation is one of the most important, chemically complex, and ubiquitous post-translational modifications in all eukaryotes. 25748215 0 63 dep Effects 0:6 arg1 Effects 0:6 Effects 0:6 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 0:131 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 63 dep Effects 0:6 arg1 study 126:130 molecular dynamics simulation study 96:130 molecular dynamics simulation study 96:130 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 0 63 dep Effects 0:6 arg1 analysis 83:90 Protein Data Bank analysis 65:90 Protein Data Bank analysis 65:90 Effects of N-glycosylation on protein conformation and dynamics: Protein Data Bank analysis and molecular dynamics simulation study. 25748215 5 64 theme protein 974:980 arg1 structure 982:990 protein structure 974:990 protein structure 974:990 Our study reveals that N-glycosylation does not induce significant changes in protein structure, but decreases protein dynamics, likely leading to an increase in protein stability. 25748215 4 65 theme N-glycans 643:651 arg1 impacts 632:638 the impacts 628:638 the impacts of N-glycans on protein structure and dynamics 628:685 In this study, the impacts of N-glycans on protein structure and dynamics are systematically investigated using an integrated computational approach of the Protein Data Bank structure analysis and atomistic molecular dynamics simulations of glycosylated and deglycosylated proteins. 26190824 4 0 theme complex 762:768 arg1 structures 779:788 less complex N-glycan structures 757:788 less complex N-glycan structures 757:788 Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. 26190824 5 1 theme N-glycosylation 866:880 arg1 sequons 882:888 N-glycosylation sequons 866:888 N-glycosylation sequons 866:888 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 5 2 theme N-glycan 1004:1011 arg1 structures 1013:1022 the N-glycan structures 1000:1022 the N-glycan structures produced 1000:1031 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 5 3 gly N-glycosylation 866:880 arg2 sequons 882:888 N-glycosylation sequons 866:888 N-glycosylation sequons 866:888 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 3 4 theme processing 606:615 arg1 trafficking 647:657 trafficking 647:657 trafficking 647:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 4 theme processing 606:615 arg1 events 617:622 other cellular processing events 591:622 other cellular processing events such as degradation or trafficking 591:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 4 theme processing 606:615 arg1 degradation 632:642 degradation 632:642 degradation 632:642 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 5 gly occupancy 347:355 arg2 sequon 364:369 the sequon 360:369 the sequon 360:369 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 3 6 from differences 576:586 arg1 trafficking 647:657 trafficking 647:657 trafficking 647:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 6 from differences 576:586 arg1 events 617:622 other cellular processing events 591:622 other cellular processing events such as degradation or trafficking 591:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 6 from differences 576:586 arg1 degradation 632:642 degradation 632:642 degradation 632:642 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 7 theme glycan 340:345 arg1 occupancy 347:355 the glycan occupancy 336:355 the glycan occupancy of the sequon in insect cells 336:385 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 3 8 theme oligosaccharyltransferase 516:540 arg1 preferences 552:562 oligosaccharyltransferase substrate preferences 516:562 oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking 516:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 9 theme Previous 183:190 arg1 studies 192:198 Previous studies 183:198 Previous studies 183:198 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 3 10 theme other 591:595 arg1 trafficking 647:657 trafficking 647:657 trafficking 647:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 10 theme other 591:595 arg1 events 617:622 other cellular processing events 591:622 other cellular processing events such as degradation or trafficking 591:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 10 theme other 591:595 arg1 degradation 632:642 degradation 632:642 degradation 632:642 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 5 11 theme glycan 962:967 arg1 occupancy 969:977 glycan occupancy 962:977 glycan occupancy 962:977 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 0 12 theme Glycosylation 61:73 arg1 Efficiency 75:84 Oligosaccharyltransferase Glycosylation Efficiency 35:84 Oligosaccharyltransferase Glycosylation Efficiency 35:84 Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. 26190824 3 13 theme cellular 597:604 arg1 trafficking 647:657 trafficking 647:657 trafficking 647:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 13 theme cellular 597:604 arg1 events 617:622 other cellular processing events 591:622 other cellular processing events such as degradation or trafficking 591:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 3 13 theme cellular 597:604 arg1 degradation 632:642 degradation 632:642 degradation 632:642 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 0 14 theme Aromatic 9:16 arg1 Sequons 18:24 Enhanced Aromatic Sequons 0:24 Enhanced Aromatic Sequons 0:24 Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. 26190824 3 15 theme in human 462:469 arg1 cells 471:475 glycan occupancy in human cells 445:475 glycan occupancy in human cells 445:475 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 0 16 theme Enhanced 0:7 arg1 Sequons 18:24 Enhanced Aromatic Sequons 0:24 Enhanced Aromatic Sequons 0:24 Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. 26190824 2 17 theme sequon 364:369 arg1 occupancy 347:355 the glycan occupancy 336:355 the glycan occupancy of the sequon in insect cells 336:385 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 1 18 theme important 135:143 arg1 role 145:148 an important role 132:148 an important role 132:148 N-Glycosylation plays an important role in protein folding and function. 26190824 4 19 theme glycan 701:706 arg1 processing 708:717 glycan processing 701:717 glycan processing 701:717 Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. 26190824 3 20 theme aromatic 411:418 arg1 residue 420:426 any aromatic residue 407:426 any aromatic residue at n-2 407:433 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 21 theme phenylalanine 219:231 arg1 residue 233:239 a phenylalanine residue 217:239 a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon 217:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 5 22 dep useful 893:898 arg1 controlling 909:919 controlling 909:919 controlling 909:919 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 5 22 dep useful 893:898 arg1 influencing 988:998 influencing 988:998 influencing the N-glycan structures produced 988:1031 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 3 23 theme glycan 445:450 arg1 cells 471:475 glycan occupancy in human cells 445:475 glycan occupancy in human cells 445:475 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 4 24 theme N-glycan 770:777 arg1 structures 779:788 less complex N-glycan structures 757:788 less complex N-glycan structures 757:788 Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. 26190824 0 25 theme Oligosaccharyltransferase 35:59 arg1 Efficiency 75:84 Oligosaccharyltransferase Glycosylation Efficiency 35:84 Oligosaccharyltransferase Glycosylation Efficiency 35:84 Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. 26190824 3 26 theme occupancy 452:460 arg1 cells 471:475 glycan occupancy in human cells 445:475 glycan occupancy in human cells 445:475 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 27 theme relative 272:279 arg1 position 263:270 the n-2 position 255:270 the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon 255:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 5 28 theme sequence 839:846 arg1 space 848:852 the sequence space 835:852 the sequence space surrounding N-glycosylation sequons 835:888 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 3 29 theme substrate 542:550 arg1 preferences 552:562 oligosaccharyltransferase substrate preferences 516:562 oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking 516:657 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 2 30 theme n-2 259:261 arg1 position 263:270 the n-2 position 255:270 the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon 255:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 1 31 theme protein 153:159 arg1 folding 161:167 protein folding 153:167 protein folding 153:167 N-Glycosylation plays an important role in protein folding and function. 26190824 2 32 theme N-glycosylation 303:317 arg1 sequon 319:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 2 33 theme insect 374:379 arg1 cells 381:385 insect cells 374:385 insect cells 374:385 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 2 34 from occupancy 347:355 arg1 cells 381:385 insect cells 374:385 insect cells 374:385 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 4 35 theme aromatic 670:677 arg1 residues 679:686 aromatic residues 670:686 aromatic residues at n-2 670:693 Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. 26190824 4 36 from n-2 691:693 arg1 residues 679:686 aromatic residues 670:686 aromatic residues at n-2 670:693 Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. 26190824 2 37 theme Asn-Xxx-Thr/Ser 287:301 arg1 sequon 319:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 26190824 0 38 theme Glycan 90:95 arg1 Homogeneity 97:107 Glycan Homogeneity 90:107 Glycan Homogeneity 90:107 Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity. 26190824 3 39 from n-2 431:433 arg1 residue 420:426 any aromatic residue 407:426 any aromatic residue at n-2 407:433 Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells and that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. 26190824 5 40 theme glycosylation 921:933 arg1 efficiency 935:944 glycosylation efficiency 921:944 glycosylation efficiency 921:944 These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced. 26190824 2 41 gly N-glycosylation 303:317 arg2 sequon 319:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 an Asn-Xxx-Thr/Ser N-glycosylation sequon 284:324 Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the glycan occupancy of the sequon in insect cells. 27869200 3 0 theme glycopeptides 534:546 arg1 spectra 523:529 the spectra 519:529 the spectra of glycopeptides 519:546 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 2 1 theme disaccharide 317:328 arg1 ions 330:333 mono- or disaccharide ions 308:333 mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans 308:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 9 2 theme glycopeptides 1522:1534 arg1 assignment 1508:1517 assignment 1508:1517 assignment of glycopeptides 1508:1534 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 8 3 theme glycosylation 1350:1362 arg1 type 1364:1367 their glycosylation type 1344:1367 their glycosylation type 1344:1367 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 2 4 theme mono- 308:312 arg1 ions 330:333 mono- or disaccharide ions 308:333 mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans 308:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 3 5 theme glycopeptides 611:623 arg1 identification 593:606 the identification 589:606 the identification of glycopeptides 589:623 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 1 6 theme protein 199:205 arg1 glycosylation 207:219 protein glycosylation 199:219 protein glycosylation 199:219 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 2 7 theme dissociation 248:259 arg1 fragmentation 267:279 Higher-energy collisional dissociation (HCD) fragmentation 222:279 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides 222:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 8 8 theme spectra 1327:1333 arg1 pre-classification 1298:1315 pre-classification 1298:1315 pre-classification of tandem spectra based on their glycosylation type 1298:1367 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 1 9 gly glycopeptides 110:122 arg2 glycopeptides 110:122 intact glycopeptides 103:122 intact glycopeptides 103:122 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 2 10 theme glycans 416:422 arg1 structure 388:396 the structure 384:396 the structure of the fragmented glycans 384:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 4 11 theme glycophorin 693:703 arg1 A 705:705 glycophorin A 693:705 glycophorin A 693:705 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 2 12 theme HCD 262:264 arg1 fragmentation 267:279 Higher-energy collisional dissociation (HCD) fragmentation 222:279 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides 222:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 3 13 gly glycopeptides 611:623 arg2 glycopeptides 611:623 glycopeptides 611:623 glycopeptides 611:623 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 3 14 theme biological 628:637 arg1 samples 639:645 biological samples 628:645 biological samples 628:645 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 6 15 theme developed 968:976 arg1 model 978:982 the developed model 964:982 the developed model 964:982 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 5 16 gly glycopeptides 937:949 arg2 glycopeptides 937:949 O-linked glycopeptides 928:949 O-linked glycopeptides 928:949 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 2 17 theme glycopeptides 284:296 arg1 fragmentation 267:279 Higher-energy collisional dissociation (HCD) fragmentation 222:279 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides 222:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 4 18 theme tryptic 728:734 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 0 19 theme N- 60:61 arg1 Identification 42:55 Identification 42:55 Identification of N- and O-linked Glycopeptides 42:88 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 2 20 contain carry 360:364 arg1 ions 350:353 oxonium ions 342:353 oxonium ions that carry information about the structure of the fragmented glycans 342:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 2 20 contain carry 360:364 arg2 information 366:376 information 366:376 information about the structure of the fragmented glycans 366:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 4 21 from spectra 655:661 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 8 22 theme N-linked 1400:1407 arg1 glycopeptides 1409:1421 N-linked glycopeptides 1400:1421 N-linked glycopeptides 1400:1421 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 0 23 theme O-linked 67:74 arg1 Glycopeptides 76:88 O-linked Glycopeptides 67:88 O-linked Glycopeptides 67:88 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 5 24 theme glycopeptides 937:949 arg1 spectra 910:916 the spectra 906:916 the spectra of N- and O-linked glycopeptides 906:949 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 4 25 theme O-linked 793:800 arg1 glycopeptides 802:814 O-linked glycopeptides 793:814 O-linked glycopeptides 793:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 7 26 theme human 1261:1265 arg1 serum 1267:1271 human serum 1261:1271 human serum 1261:1271 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 4 27 used used 749:752 arg2 spectra 655:661 Tandem spectra 648:661 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 648:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 3 28 gly glycopeptides 534:546 arg2 glycopeptides 534:546 glycopeptides 534:546 glycopeptides 534:546 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 5 29 theme Logistic 817:824 arg1 regression 826:835 Logistic regression 817:835 Logistic regression 817:835 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 6 30 theme verification 1114:1125 arg1 set 1136:1138 verification spectral set 1114:1138 verification spectral set 1114:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 7 31 theme enriched 1223:1230 arg1 glycopeptides 1232:1244 HILIC enriched glycopeptides 1217:1244 HILIC enriched glycopeptides extracted from human serum 1217:1271 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 8 32 gly glycopeptides 1409:1421 arg2 glycopeptides 1409:1421 N-linked glycopeptides 1400:1421 N-linked glycopeptides 1400:1421 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 6 33 theme ions 1103:1106 arg1 features 1079:1086 the spectral features 1066:1086 the spectral features of the oxonium ions using verification spectral set 1066:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 4 34 theme N- 786:787 arg1 database 774:781 a spectral database 763:781 a spectral database of N- and O-linked glycopeptides 763:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 0 35 theme Tandem 18:23 arg1 Spectra 30:36 Tandem Mass Spectra 18:36 Tandem Mass Spectra 18:36 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 1 36 theme intact 103:108 arg1 glycopeptides 110:122 intact glycopeptides 103:122 intact glycopeptides 103:122 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 0 37 theme Spectra 30:36 arg1 Classification 0:13 Classification 0:13 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 0:89 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 3 38 from intensity 490:498 arg1 spectra 523:529 the spectra 519:529 the spectra of glycopeptides 519:546 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 7 39 theme predictive 1183:1192 arg1 model 1194:1198 the developed predictive model 1169:1198 the developed predictive model 1169:1198 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 2 40 theme Higher-energy 222:234 arg1 fragmentation 267:279 Higher-energy collisional dissociation (HCD) fragmentation 222:279 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides 222:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 3 41 theme ions 511:514 arg1 structures 471:480 glycan structures 464:480 glycan structures 464:480 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 3 41 theme ions 511:514 arg1 intensity 490:498 the intensity 486:498 the intensity of oxonium ions in the spectra of glycopeptides 486:546 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 9 42 theme tandem 1474:1479 arg1 spectrometry 1486:1497 tandem mass spectrometry 1474:1497 tandem mass spectrometry data 1474:1502 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 4 43 theme Tandem 648:653 arg1 spectra 655:661 Tandem spectra 648:661 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 648:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 9 44 gly glycopeptides 1522:1534 arg2 glycopeptides 1522:1534 glycopeptides 1522:1534 glycopeptides 1522:1534 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 2 45 theme fragmented 405:414 arg1 glycans 416:422 the fragmented glycans 401:422 the fragmented glycans 401:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 9 46 theme spectrometry 1486:1497 arg1 data 1499:1502 tandem mass spectrometry data 1474:1502 tandem mass spectrometry data 1474:1502 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 4 47 gly glycopeptides 802:814 arg2 glycopeptides 802:814 O-linked glycopeptides 793:814 O-linked glycopeptides 793:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 1 48 gly microheterogeneity 177:194 arg1 glycosylation 207:219 protein glycosylation 199:219 protein glycosylation 199:219 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 4 49 from digests 736:742 arg1 spectra 655:661 Tandem spectra 648:661 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 648:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 4 49 from digests 736:742 arg1 glycopeptides 666:678 glycopeptides 666:678 glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 666:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 8 50 link N-linked 1400:1407 arg1 glycopeptides 1409:1421 N-linked glycopeptides 1400:1421 N-linked glycopeptides 1400:1421 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 4 51 theme A 705:705 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 2 52 gly glycopeptides 284:296 arg2 glycopeptides 284:296 glycopeptides 284:296 glycopeptides 284:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 4 53 theme ovalbumin 708:716 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 2 54 theme oxonium 342:348 arg1 ions 350:353 oxonium ions 342:353 oxonium ions that carry information about the structure of the fragmented glycans 342:422 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 1 55 theme glycosylation 207:219 arg1 microheterogeneity 177:194 the microheterogeneity 173:194 the microheterogeneity of protein glycosylation 173:219 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 8 56 theme tandem 1320:1325 arg1 spectra 1327:1333 tandem spectra 1320:1333 tandem spectra based on their glycosylation type 1320:1367 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 1 57 theme glycopeptides 110:122 arg1 Analysis 91:98 Analysis 91:98 Analysis of intact glycopeptides by mass spectrometry 91:143 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 1 58 theme mass 127:130 arg1 spectrometry 132:143 mass spectrometry 127:143 mass spectrometry 127:143 Analysis of intact glycopeptides by mass spectrometry is essential to determining the microheterogeneity of protein glycosylation. 27869200 6 59 link O-linked 1037:1044 arg1 glycopeptides 1046:1058 O-linked glycopeptides 1037:1058 O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set 1037:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 6 60 theme spectral 1070:1077 arg1 features 1079:1086 the spectral features 1066:1086 the spectral features of the oxonium ions using verification spectral set 1066:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 4 61 theme fetuin 685:690 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 5 62 theme N- 921:922 arg1 spectra 910:916 the spectra 906:916 the spectra of N- and O-linked glycopeptides 906:949 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 6 63 dep N- 1030:1031 arg1 the 1026:1028 the 1026:1028 the 1026:1028 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 4 64 theme gp120 722:726 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 0 65 theme Glycopeptides 76:88 arg1 Identification 42:55 Identification 42:55 Identification of N- and O-linked Glycopeptides 42:88 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 5 66 theme O-linked 928:935 arg1 glycopeptides 937:949 O-linked glycopeptides 928:949 O-linked glycopeptides 928:949 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 6 67 theme O-linked 1037:1044 arg1 glycopeptides 1046:1058 O-linked glycopeptides 1037:1058 O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set 1037:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 8 68 theme glycopeptides 1409:1421 arg1 identification 1382:1395 the identification 1378:1395 the identification of N-linked glycopeptides 1378:1421 The results showed that pre-classification of tandem spectra based on their glycosylation type improved the identification of N-linked glycopeptides. 27869200 4 69 gly glycopeptides 666:678 arg2 glycopeptides 666:678 glycopeptides 666:678 glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 666:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 4 69 gly glycopeptides 666:678 arg1 digests 736:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 685:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 6 70 theme spectral 1127:1134 arg1 set 1136:1138 verification spectral set 1114:1138 verification spectral set 1114:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 7 71 gly glycopeptides 1232:1244 arg2 glycopeptides 1232:1244 HILIC enriched glycopeptides 1217:1244 HILIC enriched glycopeptides extracted from human serum 1217:1271 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 7 72 theme HILIC 1217:1221 arg1 glycopeptides 1232:1244 HILIC enriched glycopeptides 1217:1244 HILIC enriched glycopeptides extracted from human serum 1217:1271 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 4 73 theme spectral 765:772 arg1 database 774:781 a spectral database 763:781 a spectral database of N- and O-linked glycopeptides 763:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 3 74 theme glycan 464:469 arg1 structures 471:480 glycan structures 464:480 glycan structures 464:480 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 0 75 theme Mass 25:28 arg1 Spectra 30:36 Tandem Mass Spectra 18:36 Tandem Mass Spectra 18:36 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 3 76 used utilized 552:559 arg2 we 431:432 we 431:432 we 431:432 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 5 77 link O-linked 928:935 arg1 glycopeptides 937:949 O-linked glycopeptides 928:949 O-linked glycopeptides 928:949 Logistic regression was applied to this database to develop model to distinguish between the spectra of N- and O-linked glycopeptides. 27869200 6 78 theme oxonium 1095:1101 arg1 ions 1103:1106 the oxonium ions 1091:1106 the oxonium ions using verification spectral set 1091:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 3 79 theme oxonium 503:509 arg1 ions 511:514 oxonium ions 503:514 oxonium ions 503:514 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 7 80 theme model 1194:1198 arg1 performance 1154:1164 the performance 1150:1164 the performance of the developed predictive model 1150:1198 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 9 81 theme developed 1428:1436 arg1 model 1438:1442 The developed model 1424:1442 The developed model 1424:1442 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 7 82 theme developed 1173:1181 arg1 model 1194:1198 the developed predictive model 1169:1198 the developed predictive model 1169:1198 Finally, the performance of the developed predictive model was evaluated in HILIC enriched glycopeptides extracted from human serum. 27869200 0 83 link O-linked 67:74 arg1 Glycopeptides 76:88 O-linked Glycopeptides 67:88 O-linked Glycopeptides 67:88 Classification of Tandem Mass Spectra for Identification of N- and O-linked Glycopeptides. 27869200 2 84 theme collisional 236:246 arg1 fragmentation 267:279 Higher-energy collisional dissociation (HCD) fragmentation 222:279 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides 222:296 Higher-energy collisional dissociation (HCD) fragmentation of glycopeptides generates mono- or disaccharide ions called oxonium ions that carry information about the structure of the fragmented glycans. 27869200 4 85 link O-linked 793:800 arg1 glycopeptides 802:814 O-linked glycopeptides 793:814 O-linked glycopeptides 793:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 3 86 from structures 471:480 arg1 spectra 523:529 the spectra 519:529 the spectra of glycopeptides 519:546 Here, we investigated the link between glycan structures and the intensity of oxonium ions in the spectra of glycopeptides and utilized this information to improve the identification of glycopeptides in biological samples. 27869200 4 87 theme glycopeptides 802:814 arg1 database 774:781 a spectral database 763:781 a spectral database of N- and O-linked glycopeptides 763:814 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 9 88 theme mass 1481:1484 arg1 spectrometry 1486:1497 tandem mass spectrometry 1474:1497 tandem mass spectrometry data 1474:1502 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 27869200 4 89 theme glycopeptides 666:678 arg1 spectra 655:661 Tandem spectra 648:661 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests 648:742 Tandem spectra of glycopeptides from fetuin, glycophorin A, ovalbumin and gp120 tryptic digests were used to build a spectral database of N- and O-linked glycopeptides. 27869200 6 90 gly glycopeptides 1046:1058 arg2 glycopeptides 1046:1058 O-linked glycopeptides 1037:1058 O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set 1037:1138 Remarkably, the developed model was found to reliably distinguish between the N- and O-linked glycopeptides using the spectral features of the oxonium ions using verification spectral set. 27869200 9 91 theme data 1499:1502 arg1 interpretation 1456:1469 interpretation 1456:1469 interpretation of tandem mass spectrometry data for assignment of glycopeptides 1456:1534 The developed model facilitates interpretation of tandem mass spectrometry data for assignment of glycopeptides. 26301455 7 0 theme significant 1396:1406 arg1 increase 1408:1415 a significant increase 1394:1415 a significant increase in the rate of Avicel and milled aspen wood hydrolysis 1394:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 2 1 theme niaD- 534:538 arg1 strain 541:546 Penicillium canescens PCA10 (niaD-) strain 505:546 Penicillium canescens PCA10 (niaD-) strain 505:546 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 1 theme niaD- 534:538 arg1 host 558:561 a fungal host 549:561 a fungal host for production of heterologous proteins 549:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 0 2 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 0:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 8 3 theme catalytic 1757:1765 arg1 machinery 1767:1775 the processive catalytic machinery 1742:1775 the processive catalytic machinery of PvCel7A 1742:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 4 4 theme variable 951:958 arg1 oligosaccharides 973:988 variable high-mannose oligosaccharides 951:988 variable high-mannose oligosaccharides 951:988 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 8 5 link N-linked 1700:1707 arg1 part 1734:1737 an important part 1721:1737 an important part of the processive catalytic machinery of PvCel7A 1721:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 8 5 link N-linked 1700:1707 arg1 glycans 1709:1715 the N-linked glycans 1696:1715 the N-linked glycans 1696:1715 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 4 6 theme mass 785:788 arg1 fingerprinting 803:816 The MALDI-TOF mass spectrometry fingerprinting 771:816 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, 771:906 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 7 7 theme N388A 1534:1538 arg1 case 1516:1519 the case 1512:1519 the case of N194A and N388A mutants 1512:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 8 8 theme 3D 1626:1627 arg1 modeling 1640:1647 3D structural modeling 1626:1647 3D structural modeling of the PvCel7A catalytic module 1626:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 7 9 theme N194A 1524:1528 arg1 case 1516:1519 the case 1512:1519 the case of N194A and N388A mutants 1512:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 8 10 theme modeling 1640:1647 arg1 data 1618:1621 data 1618:1621 data of 3D structural modeling of the PvCel7A catalytic module 1618:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 1 11 theme N-glycosylation 214:228 arg1 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 11 theme N-glycosylation 214:228 arg1 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 11 theme N-glycosylation 214:228 arg1 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 11 theme N-glycosylation 214:228 arg1 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 11 theme N-glycosylation 214:228 arg1 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 2 12 theme PCA10 527:531 arg1 strain 541:546 Penicillium canescens PCA10 (niaD-) strain 505:546 Penicillium canescens PCA10 (niaD-) strain 505:546 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 12 theme PCA10 527:531 arg1 host 558:561 a fungal host 549:561 a fungal host for production of heterologous proteins 549:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 4 13 theme peptides 821:828 arg1 fingerprinting 803:816 The MALDI-TOF mass spectrometry fingerprinting 771:816 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, 771:906 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 2 14 theme Penicillium 505:515 arg1 strain 541:546 Penicillium canescens PCA10 (niaD-) strain 505:546 Penicillium canescens PCA10 (niaD-) strain 505:546 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 14 theme Penicillium 505:515 arg1 host 558:561 a fungal host 549:561 a fungal host for production of heterologous proteins 549:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 0 15 from effect 104:109 arg1 activity 125:132 the enzyme activity 114:132 the enzyme activity 114:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 15 from effect 104:109 arg1 verruculosum 83:94 Penicillium verruculosum 71:94 Penicillium verruculosum 71:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 8 16 theme catalytic 1664:1672 arg1 module 1674:1679 the PvCel7A catalytic module 1652:1679 the PvCel7A catalytic module 1652:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 7 17 theme digestion 1493:1501 arg1 rates 1503:1507 the substrate digestion rates 1479:1507 the substrate digestion rates in the case of N194A and N388A mutants 1479:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 7 17 theme digestion 1493:1501 arg1 lower 1561:1565 lower 1561:1565 lower 1561:1565 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 5 18 theme thermostability 1199:1213 arg1 pH-optimum 1157:1166 pH-optimum 1157:1166 pH-optimum of PvCel7A activity and enzyme thermostability 1157:1213 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 1 19 from verruculosum 172:183 arg1 I 153:153 Cellobiohydrolase I 135:153 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) 135:193 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 2 20 theme mutant 439:444 arg1 forms 446:450 its mutant forms 435:450 its mutant forms 435:450 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 6 21 from activity 1303:1310 arg1 Avicel 1288:1293 Avicel 1288:1293 Avicel 1288:1293 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 2 22 theme Ala 469:471 arg1 substitutions 473:485 Asn to Ala substitutions 462:485 Asn to Ala substitutions 462:485 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 5 23 theme PvCel7A 1171:1177 arg1 activity 1179:1186 PvCel7A activity 1171:1186 PvCel7A activity 1171:1186 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 3 24 theme mutant 746:751 arg1 expression 726:735 the expression 722:735 the expression of N430A mutant 722:751 The rPvCel7A-wt and N45A, N194A, N388A mutants were successfully expressed and purified for characterization, whereas the expression of N430A mutant was not achieved. 26301455 8 25 theme PvCel7A 1780:1786 arg1 machinery 1767:1775 the processive catalytic machinery 1742:1775 the processive catalytic machinery of PvCel7A 1742:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 8 26 theme N-linked 1700:1707 arg1 part 1734:1737 an important part 1721:1737 an important part of the processive catalytic machinery of PvCel7A 1721:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 8 26 theme N-linked 1700:1707 arg1 glycans 1709:1715 the N-linked glycans 1696:1715 the N-linked glycans 1696:1715 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 7 27 theme wood 1456:1459 arg1 hydrolysis 1461:1470 milled aspen wood hydrolysis 1443:1470 milled aspen wood hydrolysis 1443:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 2 28 theme Asn 462:464 arg1 substitutions 473:485 Asn to Ala substitutions 462:485 Asn to Ala substitutions 462:485 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 6 29 theme enzyme 1259:1264 arg1 ability 1277:1283 the enzyme adsorption ability 1255:1283 the enzyme adsorption ability on Avicel 1255:1293 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 4 30 theme high-mannose 960:971 arg1 oligosaccharides 973:988 variable high-mannose oligosaccharides 951:988 variable high-mannose oligosaccharides 951:988 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 4 31 link N-linked 924:931 arg1 glycans 933:939 the N-linked glycans 920:939 the N-linked glycans 920:939 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 4 32 theme GlcNAc 1106:1111 arg1 residue 1113:1119 a single GlcNAc residue 1097:1119 a single GlcNAc residue 1097:1119 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 0 33 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 0:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 8 34 theme important 1724:1732 arg1 part 1734:1737 an important part 1721:1737 an important part of the processive catalytic machinery of PvCel7A 1721:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 8 34 theme important 1724:1732 arg1 glycans 1709:1715 the N-linked glycans 1696:1715 the N-linked glycans 1696:1715 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 2 35 theme wild 408:411 arg1 type 413:416 the wild type 404:416 the wild type (wt) PvCel7A 404:429 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 35 theme wild 408:411 arg1 wt 419:420 wt 419:420 wt 419:420 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 36 theme proteins 594:601 arg1 production 567:576 production 567:576 production of heterologous proteins 567:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 4 37 theme N-linked 924:931 arg1 glycans 933:939 the N-linked glycans 920:939 the N-linked glycans 920:939 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 0 38 theme cellobiohydrolase 38:54 arg1 effect 104:109 its effect 100:109 its effect on the enzyme activity 100:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 38 theme cellobiohydrolase 38:54 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 0:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 39 gly glycosylation 9:21 arg1 activity 125:132 the enzyme activity 114:132 the enzyme activity 114:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 39 gly glycosylation 9:21 arg1 cellobiohydrolase 38:54 recombinant cellobiohydrolase I 26:56 recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 26:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 39 gly glycosylation 9:21 arg1 Cel7A 59:63 Cel7A 59:63 Cel7A 59:63 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 4 40 theme enzymatic 1027:1035 arg1 trimming 1037:1044 their sequential enzymatic trimming 1010:1044 their sequential enzymatic trimming 1010:1044 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 3 41 theme rPvCel7A-wt 608:618 arg1 mutants 643:649 The rPvCel7A-wt and N45A, N194A, N388A mutants 604:649 The rPvCel7A-wt and N45A, N194A, N388A mutants 604:649 The rPvCel7A-wt and N45A, N194A, N388A mutants were successfully expressed and purified for characterization, whereas the expression of N430A mutant was not achieved. 26301455 1 42 contain has 195:197 arg2 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 42 contain has 195:197 arg2 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 42 contain has 195:197 arg1 I 153:153 Cellobiohydrolase I 135:153 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) 135:193 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 42 contain has 195:197 arg2 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 42 contain has 195:197 arg2 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 42 contain has 195:197 arg2 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 5 43 theme notable 1139:1145 arg1 effect 1147:1152 no notable effect 1136:1152 no notable effect 1136:1152 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 4 44 theme spectrometry 790:801 arg1 fingerprinting 803:816 The MALDI-TOF mass spectrometry fingerprinting 771:816 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, 771:906 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 8 45 theme processive 1746:1755 arg1 machinery 1767:1775 the processive catalytic machinery 1742:1775 the processive catalytic machinery of PvCel7A 1742:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 2 46 dep substitutions 473:485 arg1 to 466:467 to 466:467 to 466:467 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 7 47 theme N45A 1373:1376 arg1 mutation 1378:1385 the N45A mutation 1369:1385 the N45A mutation 1369:1385 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 5 48 contain had 1132:1134 arg2 effect 1147:1152 no notable effect 1136:1152 no notable effect 1136:1152 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 5 48 contain had 1132:1134 arg1 Mutations 1122:1130 Mutations 1122:1130 Mutations 1122:1130 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 8 49 theme machinery 1767:1775 arg1 part 1734:1737 an important part 1721:1737 an important part of the processive catalytic machinery of PvCel7A 1721:1786 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 8 49 theme machinery 1767:1775 arg1 glycans 1709:1715 the N-linked glycans 1696:1715 the N-linked glycans 1696:1715 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 2 50 theme fungal 551:556 arg1 strain 541:546 Penicillium canescens PCA10 (niaD-) strain 505:546 Penicillium canescens PCA10 (niaD-) strain 505:546 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 50 theme fungal 551:556 arg1 host 558:561 a fungal host 549:561 a fungal host for production of heterologous proteins 549:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 3 51 theme N388A 637:641 arg1 mutants 643:649 The rPvCel7A-wt and N45A, N194A, N388A mutants 604:649 The rPvCel7A-wt and N45A, N194A, N388A mutants 604:649 The rPvCel7A-wt and N45A, N194A, N388A mutants were successfully expressed and purified for characterization, whereas the expression of N430A mutant was not achieved. 26301455 1 52 gly N-glycosylation 214:228 arg2 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 module 253:258 its catalytic module 239:258 its catalytic module 239:258 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 52 gly N-glycosylation 214:228 arg2 four 199:202 four 199:202 four 199:202 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 4 53 theme MALDI-TOF 775:783 arg1 fingerprinting 803:816 The MALDI-TOF mass spectrometry fingerprinting 771:816 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, 771:906 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 0 54 theme enzyme 118:123 arg1 activity 125:132 the enzyme activity 114:132 the enzyme activity 114:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 55 from verruculosum 83:94 arg1 effect 104:109 its effect 100:109 its effect on the enzyme activity 100:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 55 from verruculosum 83:94 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 0:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 55 from verruculosum 83:94 arg1 cellobiohydrolase 38:54 recombinant cellobiohydrolase I 26:56 recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 26:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 55 from verruculosum 83:94 arg1 Cel7A 59:63 Cel7A 59:63 Cel7A 59:63 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 4 56 theme formula 1064:1070 arg1 Man 1073:1075 the formula (Man)0-13 1060:1080 the formula (Man)0-13 (GlcNAc)2 1060:1090 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 1 57 from module 253:258 arg1 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 57 from module 253:258 arg1 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 57 from module 253:258 arg1 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 57 from module 253:258 arg1 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 57 from module 253:258 arg1 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 58 theme potential 204:212 arg1 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 58 theme potential 204:212 arg1 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 58 theme potential 204:212 arg1 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 58 theme potential 204:212 arg1 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 58 theme potential 204:212 arg1 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 4 59 theme digestion 855:863 arg1 result 845:850 a result 843:850 a result of digestion of rPvCel7A forms with specific proteases 843:905 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 1 60 dep sites 230:234 arg1 Asn388 276:281 Asn388 276:281 Asn388 276:281 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 60 dep sites 230:234 arg1 Asn430 288:293 Asn430 288:293 Asn430 288:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 60 dep sites 230:234 arg1 Asn194 268:273 Asn194 268:273 Asn194 268:273 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 60 dep sites 230:234 arg1 Asn45 261:265 Asn45 261:265 Asn45 261:265 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 1 60 dep sites 230:234 arg1 sites 230:234 four potential N-glycosylation sites 199:234 four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430 199:293 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 4 61 theme forms 877:881 arg1 digestion 855:863 digestion 855:863 digestion of rPvCel7A forms with specific proteases 855:905 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 8 62 theme structural 1629:1638 arg1 modeling 1640:1647 3D structural modeling 1626:1647 3D structural modeling of the PvCel7A catalytic module 1626:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 4 63 theme single 1099:1104 arg1 residue 1113:1119 a single GlcNAc residue 1097:1119 a single GlcNAc residue 1097:1119 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 5 64 theme activity 1179:1186 arg1 pH-optimum 1157:1166 pH-optimum 1157:1166 pH-optimum of PvCel7A activity and enzyme thermostability 1157:1213 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 7 65 theme substrate 1483:1491 arg1 rates 1503:1507 the substrate digestion rates 1479:1507 the substrate digestion rates in the case of N194A and N388A mutants 1479:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 7 65 theme substrate 1483:1491 arg1 lower 1561:1565 lower 1561:1565 lower 1561:1565 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 4 66 theme GlcNAc 1083:1088 arg1 Man 1073:1075 the formula (Man)0-13 1060:1080 the formula (Man)0-13 (GlcNAc)2 1060:1090 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 5 67 theme enzyme 1192:1197 arg1 thermostability 1199:1213 enzyme thermostability 1192:1213 enzyme thermostability 1192:1213 Mutations had no notable effect on pH-optimum of PvCel7A activity and enzyme thermostability. 26301455 3 68 theme N430A 740:744 arg1 mutant 746:751 N430A mutant 740:751 N430A mutant 740:751 The rPvCel7A-wt and N45A, N194A, N388A mutants were successfully expressed and purified for characterization, whereas the expression of N430A mutant was not achieved. 26301455 8 69 theme PvCel7A 1656:1662 arg1 module 1674:1679 the PvCel7A catalytic module 1652:1679 the PvCel7A catalytic module 1652:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 1 70 theme Cellobiohydrolase 135:151 arg1 I 153:153 Cellobiohydrolase I 135:153 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) 135:193 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 26301455 4 71 theme specific 888:895 arg1 proteases 897:905 specific proteases 888:905 specific proteases 888:905 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 7 72 dep N194A 1524:1528 arg1 mutants 1540:1546 mutants 1540:1546 mutants 1540:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 7 73 from rates 1503:1507 arg1 case 1516:1519 the case 1512:1519 the case of N194A and N388A mutants 1512:1546 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 8 74 theme module 1674:1679 arg1 modeling 1640:1647 3D structural modeling 1626:1647 3D structural modeling of the PvCel7A catalytic module 1626:1679 These data, together with data of 3D structural modeling of the PvCel7A catalytic module, indicate that the N-linked glycans are an important part of the processive catalytic machinery of PvCel7A. 26301455 7 75 theme hydrolysis 1461:1470 arg1 rate 1424:1427 the rate 1420:1427 the rate of Avicel and milled aspen wood hydrolysis 1420:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 0 76 from glycosylation 9:21 arg1 activity 125:132 the enzyme activity 114:132 the enzyme activity 114:132 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 76 from glycosylation 9:21 arg1 verruculosum 83:94 Penicillium verruculosum 71:94 Penicillium verruculosum 71:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 2 77 theme heterologous 581:592 arg1 proteins 594:601 heterologous proteins 581:601 heterologous proteins 581:601 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 7 78 theme aspen 1450:1454 arg1 hydrolysis 1461:1470 milled aspen wood hydrolysis 1443:1470 milled aspen wood hydrolysis 1443:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 4 79 theme rPvCel7A 868:875 arg1 forms 877:881 rPvCel7A forms 868:881 rPvCel7A forms 868:881 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 6 80 theme adsorption 1266:1275 arg1 ability 1277:1283 the enzyme adsorption ability 1255:1283 the enzyme adsorption ability on Avicel 1255:1293 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 4 81 with digestion 855:863 arg1 proteases 897:905 specific proteases 888:905 specific proteases 888:905 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 4 82 theme residue 1113:1119 arg1 oligosaccharides 973:988 variable high-mannose oligosaccharides 951:988 variable high-mannose oligosaccharides 951:988 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 4 82 theme residue 1113:1119 arg1 products 998:1005 the products 994:1005 the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue 994:1119 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 2 83 theme other 372:376 arg1 properties 378:387 other properties 372:387 other properties 372:387 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 6 84 from ability 1277:1283 arg1 Avicel 1288:1293 Avicel 1288:1293 Avicel 1288:1293 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 2 85 dep Penicillium 505:515 arg1 canescens 517:525 canescens 517:525 canescens 517:525 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 86 theme type 413:416 arg1 PvCel7A 423:429 the wild type (wt) PvCel7A 404:429 the wild type (wt) PvCel7A 404:429 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 0 87 theme recombinant 26:36 arg1 cellobiohydrolase 38:54 recombinant cellobiohydrolase I 26:56 recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum 26:94 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 0 87 theme recombinant 26:36 arg1 Cel7A 59:63 Cel7A 59:63 Cel7A 59:63 N-linked glycosylation of recombinant cellobiohydrolase I (Cel7A) from Penicillium verruculosum and its effect on the enzyme activity. 26301455 2 88 theme enzyme 396:401 arg1 activity 359:366 activity 359:366 activity 359:366 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 88 theme enzyme 396:401 arg1 properties 378:387 other properties 372:387 other properties 372:387 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 6 89 theme synthetic 1332:1340 arg1 substrates 1342:1351 natural and synthetic substrates 1320:1351 natural and synthetic substrates 1320:1351 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 7 90 theme milled 1443:1448 arg1 hydrolysis 1461:1470 milled aspen wood hydrolysis 1443:1470 milled aspen wood hydrolysis 1443:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 6 91 theme natural 1320:1326 arg1 substrates 1342:1351 natural and synthetic substrates 1320:1351 natural and synthetic substrates 1320:1351 However, the mutations influenced both the enzyme adsorption ability on Avicel and its activity against natural and synthetic substrates. 26301455 7 92 theme Avicel 1432:1437 arg1 rate 1424:1427 the rate 1420:1427 the rate of Avicel and milled aspen wood hydrolysis 1420:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 4 93 theme sequential 1016:1025 arg1 trimming 1037:1044 their sequential enzymatic trimming 1010:1044 their sequential enzymatic trimming 1010:1044 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 2 94 dep activity 359:366 arg1 the 355:357 the 355:357 the 355:357 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 95 contain carrying 453:460 arg1 forms 446:450 its mutant forms 435:450 its mutant forms 435:450 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 2 95 contain carrying 453:460 arg2 substitutions 473:485 Asn to Ala substitutions 462:485 Asn to Ala substitutions 462:485 In order to investigate how the N-glycosylation influences the activity and other properties of the enzyme, the wild type (wt) PvCel7A and its mutant forms, carrying Asn to Ala substitutions, were cloned into Penicillium canescens PCA10 (niaD-) strain, a fungal host for production of heterologous proteins. 26301455 7 96 from increase 1408:1415 arg1 rate 1424:1427 the rate 1420:1427 the rate of Avicel and milled aspen wood hydrolysis 1420:1470 In particular, the N45A mutation led to a significant increase in the rate of Avicel and milled aspen wood hydrolysis, while the substrate digestion rates in the case of N194A and N388A mutants were notably lower relative to rPvCel7A-wt. 26301455 4 97 theme trimming 1037:1044 arg1 oligosaccharides 973:988 variable high-mannose oligosaccharides 951:988 variable high-mannose oligosaccharides 951:988 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 4 97 theme trimming 1037:1044 arg1 products 998:1005 the products 994:1005 the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue 994:1119 The MALDI-TOF mass spectrometry fingerprinting of peptides, obtained as a result of digestion of rPvCel7A forms with specific proteases, showed that the N-linked glycans represent variable high-mannose oligosaccharides and the products of their sequential enzymatic trimming, according to the formula (Man)0-13 (GlcNAc)2 , or a single GlcNAc residue. 26301455 1 98 theme catalytic 243:251 arg1 module 253:258 its catalytic module 239:258 its catalytic module 239:258 Cellobiohydrolase I from Penicillium verruculosum (PvCel7A) has four potential N-glycosylation sites at its catalytic module: Asn45, Asn194, Asn388, and Asn430. 25190359 7 0 theme N-glycosylation 1329:1343 arg1 phenotype 1345:1353 the CF sputum N-glycosylation phenotype 1315:1353 the CF sputum N-glycosylation phenotype 1315:1353 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 6 1 theme microbial 1067:1075 arg1 flora 1077:1081 microbial flora 1067:1081 microbial flora 1067:1081 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 6 1 theme microbial 1067:1075 arg1 genotypes 1040:1048 different genotypes 1030:1048 different genotypes 1030:1048 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 7 2 theme CF 1319:1320 arg1 phenotype 1345:1353 the CF sputum N-glycosylation phenotype 1315:1353 the CF sputum N-glycosylation phenotype 1315:1353 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 3 3 theme chromatography-negative 614:636 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 1 4 theme CF 275:276 arg1 sputum 278:283 CF sputum 275:283 CF sputum 275:283 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 7 5 with paucimannose 1360:1371 arg1 over-sialylation/fucosylation 1391:1419 simultaneous over-sialylation/fucosylation 1378:1419 simultaneous over-sialylation/fucosylation 1378:1419 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 5 with paucimannose 1360:1371 arg1 GlcNAcylation 1441:1453 under-bisecting GlcNAcylation 1425:1453 under-bisecting GlcNAcylation 1425:1453 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 10 6 theme structural 1870:1879 arg1 information 1890:1900 new structural N-glycan information 1866:1900 new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract 1866:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 3 7 theme graphitized 588:598 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 9 8 theme secondary 1746:1754 arg1 effect 1756:1761 a secondary effect 1744:1761 a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum 1744:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 8 theme secondary 1746:1754 arg1 N-glycosylation 1691:1705 protein N-glycosylation 1683:1705 protein N-glycosylation 1683:1705 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 8 9 theme CF 1513:1514 arg1 patients 1516:1523 CF patients 1513:1523 CF patients 1513:1523 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 6 10 theme longitudinal 1137:1148 arg1 variation 1150:1158 little interpatient and longitudinal variation 1113:1158 little interpatient and longitudinal variation 1113:1158 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 4 11 theme topology 734:741 arg1 isomers 755:761 The composition, topology and linkage isomers 717:761 The composition, topology and linkage isomers of 68 N-glycans 717:777 The composition, topology and linkage isomers of 68 N-glycans were characterized and relatively quantified. 25190359 3 12 theme mass 649:652 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 9 13 theme infection 1776:1784 arg1 effect 1756:1761 a secondary effect 1744:1761 a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum 1744:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 13 theme infection 1776:1784 arg1 N-glycosylation 1691:1705 protein N-glycosylation 1683:1705 protein N-glycosylation 1683:1705 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 6 14 theme interpatient 1120:1131 arg1 variation 1150:1158 little interpatient and longitudinal variation 1113:1158 little interpatient and longitudinal variation 1113:1158 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 4 15 theme linkage 747:753 arg1 isomers 755:761 The composition, topology and linkage isomers 717:761 The composition, topology and linkage isomers of 68 N-glycans 717:777 The composition, topology and linkage isomers of 68 N-glycans were characterized and relatively quantified. 25190359 10 16 theme molecular 1946:1954 arg1 environment 1956:1966 the complex cellular and molecular environment 1921:1966 the complex cellular and molecular environment of the CF affected respiratory tract 1921:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 5 17 theme structural 835:844 arg1 features 846:853 Recurring structural features 825:853 Recurring structural features in all sputum N-glycomes 825:878 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 5 18 from features 846:853 arg1 N-glycomes 869:878 all sputum N-glycomes 858:878 all sputum N-glycomes 858:878 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 3 19 theme sputum 700:705 arg1 proteins 707:714 sputum proteins 700:714 sputum proteins 700:714 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 9 20 theme inflammation 1790:1801 arg1 effect 1756:1761 a secondary effect 1744:1761 a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum 1744:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 20 theme inflammation 1790:1801 arg1 N-glycosylation 1691:1705 protein N-glycosylation 1683:1705 protein N-glycosylation 1683:1705 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 10 21 theme cellular 1933:1940 arg1 environment 1956:1966 the complex cellular and molecular environment 1921:1966 the complex cellular and molecular environment of the CF affected respiratory tract 1921:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 7 22 theme Comparative 1180:1190 arg1 analysis 1202:1209 Comparative N-glycome analysis 1180:1209 Comparative N-glycome analysis between inter-patient group 1180:1237 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 6 23 theme sputum 1088:1093 arg1 N-glycomes 1095:1104 the sputum N-glycomes 1084:1104 the sputum N-glycomes 1084:1104 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 8 24 theme paucimannose 1566:1577 arg1 N-glycans 1579:1587 paucimannose N-glycans 1566:1587 paucimannose N-glycans 1566:1587 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 9 25 theme CF 1819:1820 arg1 pathogenesis 1822:1833 the CF pathogenesis 1815:1833 the CF pathogenesis 1815:1833 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 8 26 theme relative 1589:1596 arg1 abundance 1553:1561 abundance 1553:1561 abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals 1553:1646 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 0 27 theme N-glycosylation 61:75 arg1 phenotype 77:85 the sputum N-glycosylation phenotype 50:85 the sputum N-glycosylation phenotype 50:85 Cystic fibrosis and bacterial colonization define the sputum N-glycosylation phenotype. 25190359 2 28 theme pulmonary 529:537 arg1 conditions 539:548 other pulmonary conditions 523:548 other pulmonary conditions 523:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 8 29 theme pathogen-infected/colonized 1601:1627 arg1 individuals 1636:1646 pathogen-infected/colonized non-CF individuals 1601:1646 pathogen-infected/colonized non-CF individuals 1601:1646 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 7 30 theme N-glycans 1473:1481 arg1 over-sialylation/fucosylation 1391:1419 simultaneous over-sialylation/fucosylation 1378:1419 simultaneous over-sialylation/fucosylation 1378:1419 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 30 theme N-glycans 1473:1481 arg1 GlcNAcylation 1441:1453 under-bisecting GlcNAcylation 1425:1453 under-bisecting GlcNAcylation 1425:1453 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 10 31 theme affected 1978:1985 arg1 tract 1999:2003 the CF affected respiratory tract 1971:2003 the CF affected respiratory tract 1971:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 5 32 theme sputum 862:867 arg1 N-glycomes 869:878 all sputum N-glycomes 858:878 all sputum N-glycomes 858:878 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 1 33 gly N-glycosylation 256:270 arg1 sputum 278:283 CF sputum 275:283 CF sputum 275:283 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 2 34 theme pathogen-infected/colonized 461:487 arg1 individuals 496:506 two pathogen-infected/colonized non-CF individuals 457:506 two pathogen-infected/colonized non-CF individuals 457:506 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 0 35 theme Cystic 0:5 arg1 fibrosis 7:14 Cystic fibrosis 0:14 Cystic fibrosis 0:14 Cystic fibrosis and bacterial colonization define the sputum N-glycosylation phenotype. 25190359 1 36 theme sputum 278:283 arg1 N-glycosylation 256:270 protein N-glycosylation 248:270 protein N-glycosylation of CF sputum 248:283 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 1 37 theme sputum 122:127 arg1 O-glycosylation 103:117 mucin O-glycosylation 97:117 mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) 97:180 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 1 38 theme cystic 161:166 arg1 fibrosis 168:175 cystic fibrosis 161:175 cystic fibrosis (CF) 161:180 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 1 38 theme cystic 161:166 arg1 CF 178:179 CF 178:179 CF 178:179 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 7 39 theme under-bisecting 1425:1439 arg1 GlcNAcylation 1441:1453 under-bisecting GlcNAcylation 1425:1453 under-bisecting GlcNAcylation 1425:1453 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 10 40 theme tract 1999:2003 arg1 environment 1956:1966 the complex cellular and molecular environment 1921:1966 the complex cellular and molecular environment of the CF affected respiratory tract 1921:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 5 41 theme α1,6-core 912:920 arg1 fucosylation 922:933 α1,6-core fucosylation 912:933 α1,6-core fucosylation 912:933 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 6 42 theme different 1030:1038 arg1 gender 1056:1061 gender 1056:1061 gender 1056:1061 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 6 42 theme different 1030:1038 arg1 flora 1077:1081 microbial flora 1067:1081 microbial flora 1067:1081 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 6 42 theme different 1030:1038 arg1 age 1051:1053 age 1051:1053 age 1051:1053 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 6 42 theme different 1030:1038 arg1 genotypes 1040:1048 different genotypes 1030:1048 different genotypes 1030:1048 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 9 43 theme protein 1683:1689 arg1 effect 1756:1761 a secondary effect 1744:1761 a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum 1744:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 43 theme protein 1683:1689 arg1 N-glycosylation 1691:1705 protein N-glycosylation 1683:1705 protein N-glycosylation 1683:1705 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 44 from effect 1756:1761 arg1 sputum 1838:1843 sputum 1838:1843 sputum 1838:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 10 45 theme new 1866:1868 arg1 information 1890:1900 new structural N-glycan information 1866:1900 new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract 1866:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 10 46 theme respiratory 1987:1997 arg1 tract 1999:2003 the CF affected respiratory tract 1971:2003 the CF affected respiratory tract 1971:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 7 47 gly over-sialylation/fucosylation 1391:1419 arg1 paucimannose 1360:1371 paucimannose 1360:1371 paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans 1360:1481 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 47 gly over-sialylation/fucosylation 1391:1419 arg1 N-glycans 1473:1481 complex/hybrid N-glycans 1458:1481 complex/hybrid N-glycans 1458:1481 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 48 theme sputum 1322:1327 arg1 phenotype 1345:1353 the CF sputum N-glycosylation phenotype 1315:1353 the CF sputum N-glycosylation phenotype 1315:1353 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 2 49 theme soluble 377:383 arg1 proteins 385:392 soluble proteins 377:392 soluble proteins 377:392 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 1 50 theme protein 248:254 arg1 N-glycosylation 256:270 protein N-glycosylation 248:270 protein N-glycosylation of CF sputum 248:283 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 3 51 theme carbon-liquid 600:612 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 2 52 from N-glycome 364:372 arg1 sputum 397:402 sputum 397:402 sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions 397:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 10 53 theme N-glycan 1881:1888 arg1 information 1890:1900 new structural N-glycan information 1866:1900 new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract 1866:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 3 54 theme porous 581:586 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 4 55 theme N-glycans 769:777 arg1 isomers 755:761 The composition, topology and linkage isomers 717:761 The composition, topology and linkage isomers of 68 N-glycans 717:777 The composition, topology and linkage isomers of 68 N-glycans were characterized and relatively quantified. 25190359 10 56 theme complex 1925:1931 arg1 environment 1956:1966 the complex cellular and molecular environment 1921:1966 the complex cellular and molecular environment of the CF affected respiratory tract 1921:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 3 57 theme ion-tandem 638:647 arg1 spectrometry 654:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry 581:665 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 9 58 theme bacterial 1766:1774 arg1 infection 1776:1784 bacterial infection 1766:1784 bacterial infection 1766:1784 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 7 59 theme lung 1253:1256 arg1 infection/colonization 1258:1279 lung infection/colonization 1253:1279 lung infection/colonization 1253:1279 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 6 60 theme little 1113:1118 arg1 variation 1150:1158 little interpatient and longitudinal variation 1113:1158 little interpatient and longitudinal variation 1113:1158 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 7 61 theme complex/hybrid 1458:1471 arg1 N-glycans 1473:1481 complex/hybrid N-glycans 1458:1481 complex/hybrid N-glycans 1458:1481 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 3 62 theme enzymatic 677:685 arg1 release 687:693 enzymatic release 677:693 enzymatic release from sputum proteins 677:714 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 7 63 theme simultaneous 1378:1389 arg1 over-sialylation/fucosylation 1391:1419 simultaneous over-sialylation/fucosylation 1378:1419 simultaneous over-sialylation/fucosylation 1378:1419 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 64 theme inter-patient 1219:1231 arg1 group 1233:1237 inter-patient group 1219:1237 inter-patient group 1219:1237 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 7 65 theme N-glycome 1192:1200 arg1 analysis 1202:1209 Comparative N-glycome analysis 1180:1209 Comparative N-glycome analysis between inter-patient group 1180:1237 Comparative N-glycome analysis between inter-patient group revealed that lung infection/colonization, in general, extensively enriches the CF sputum N-glycosylation phenotype with paucimannose with simultaneous over-sialylation/fucosylation and under-bisecting GlcNAcylation of complex/hybrid N-glycans. 25190359 9 66 theme pathogenesis 1822:1833 arg1 effect 1756:1761 a secondary effect 1744:1761 a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum 1744:1843 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 9 66 theme pathogenesis 1822:1833 arg1 N-glycosylation 1691:1705 protein N-glycosylation 1683:1705 protein N-glycosylation 1683:1705 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 8 67 theme N-glycans 1579:1587 arg1 abundance 1553:1561 abundance 1553:1561 abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals 1553:1646 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 5 68 theme terminal 885:892 arg1 α2,6-sialylation 894:909 terminal α2,6-sialylation 885:909 terminal α2,6-sialylation 885:909 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 8 69 from patients 1516:1523 arg1 sputum 1501:1506 the sputum 1497:1506 the sputum from CF patients 1497:1523 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 2 70 theme other 523:527 arg1 conditions 539:548 other pulmonary conditions 523:548 other pulmonary conditions 523:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 1 71 gly O-glycosylation 103:117 arg1 sputum 122:127 sputum 122:127 sputum 122:127 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 6 72 theme CF 1167:1168 arg1 patients 1170:1177 CF patients 1167:1177 CF patients 1167:1177 Despite covering different genotypes, age, gender and microbial flora, the sputum N-glycomes showed little interpatient and longitudinal variation within CF patients. 25190359 8 73 theme non-CF 1629:1634 arg1 individuals 1636:1646 pathogen-infected/colonized non-CF individuals 1601:1646 pathogen-infected/colonized non-CF individuals 1601:1646 In contrast, the sputum from CF patients had only slightly increased abundance of paucimannose N-glycans relative to pathogen-infected/colonized non-CF individuals. 25190359 3 74 from proteins 707:714 arg1 release 687:693 enzymatic release 677:693 enzymatic release from sputum proteins 677:714 N-Glycans were profiled using porous graphitized carbon-liquid chromatography-negative ion-tandem mass spectrometry following enzymatic release from sputum proteins. 25190359 10 75 theme CF 1975:1976 arg1 tract 1999:2003 the CF affected respiratory tract 1971:2003 the CF affected respiratory tract 1971:2003 This study provides new structural N-glycan information to help understand the complex cellular and molecular environment of the CF affected respiratory tract. 25190359 5 76 theme β1,4-bisecting 936:949 arg1 GlcNAcylation 951:963 β1,4-bisecting GlcNAcylation 936:963 β1,4-bisecting GlcNAcylation 936:963 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 1 77 theme mucin 97:101 arg1 O-glycosylation 103:117 mucin O-glycosylation 97:117 mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) 97:180 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 5 78 theme Recurring 825:833 arg1 features 846:853 Recurring structural features 825:853 Recurring structural features in all sputum N-glycomes 825:878 Recurring structural features in all sputum N-glycomes were terminal α2,6-sialylation, α1,6-core fucosylation, β1,4-bisecting GlcNAcylation and compositions indicating paucimannosylation. 25190359 0 79 theme bacterial 20:28 arg1 colonization 30:41 bacterial colonization 20:41 bacterial colonization 20:41 Cystic fibrosis and bacterial colonization define the sputum N-glycosylation phenotype. 25190359 1 80 from individuals 134:144 arg1 O-glycosylation 103:117 mucin O-glycosylation 97:117 mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) 97:180 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 2 81 theme CF 422:423 arg1 patients 425:432 five CF patients 417:432 five CF patients 417:432 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 0 82 theme sputum 54:59 arg1 phenotype 77:85 the sputum N-glycosylation phenotype 50:85 the sputum N-glycosylation phenotype 50:85 Cystic fibrosis and bacterial colonization define the sputum N-glycosylation phenotype. 25190359 9 83 theme mucin 1660:1664 arg1 O-glycosylation 1666:1680 mucin O-glycosylation 1660:1680 mucin O-glycosylation 1660:1680 Similar to mucin O-glycosylation, protein N-glycosylation appears to be regulated primarily as a secondary effect of bacterial infection and inflammation rather than the CF pathogenesis in sputum. 25190359 2 84 theme non-CF 489:494 arg1 individuals 496:506 two pathogen-infected/colonized non-CF individuals 457:506 two pathogen-infected/colonized non-CF individuals 457:506 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 2 85 theme first 358:362 arg1 N-glycome 364:372 the first N-glycome 354:372 the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions 354:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 1 86 theme unaffected 223:232 arg1 counterparts 234:245 their unaffected counterparts 217:245 their unaffected counterparts 217:245 Although mucin O-glycosylation of sputum from individuals suffering from cystic fibrosis (CF) is known to be altered relative to their unaffected counterparts, protein N-glycosylation of CF sputum remains structurally and functionally under-characterized. 25190359 2 87 attach derived 404:410 arg2 sputum 397:402 sputum 397:402 sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions 397:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 2 87 attach derived 404:410 arg1 pathogen-free 439:451 pathogen-free 439:451 pathogen-free 439:451 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 2 87 attach derived 404:410 arg1 patients 425:432 five CF patients 417:432 five CF patients 417:432 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 2 87 attach derived 404:410 arg1 individuals 496:506 two pathogen-infected/colonized non-CF individuals 457:506 two pathogen-infected/colonized non-CF individuals 457:506 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25190359 4 88 theme composition 721:731 arg1 isomers 755:761 The composition, topology and linkage isomers 717:761 The composition, topology and linkage isomers of 68 N-glycans 717:777 The composition, topology and linkage isomers of 68 N-glycans were characterized and relatively quantified. 25190359 2 89 theme proteins 385:392 arg1 N-glycome 364:372 the first N-glycome 354:372 the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions 354:548 We report the first N-glycome of soluble proteins in sputum derived from five CF patients, two pathogen-free and two pathogen-infected/colonized non-CF individuals suffering from other pulmonary conditions. 25730103 5 0 theme glycomic 678:685 arg1 markers 687:693 glycomic markers 678:693 glycomic markers 678:693 In particular, glycomic markers have often proven to be useful in distinguishing cancers from precancerous conditions. 25730103 5 1 theme precancerous 757:768 arg1 conditions 770:779 precancerous conditions 757:779 precancerous conditions 757:779 In particular, glycomic markers have often proven to be useful in distinguishing cancers from precancerous conditions. 25730103 4 2 theme various 646:652 arg1 cancers 654:660 various cancers 646:660 various cancers 646:660 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 8 3 theme larger 1263:1268 arg1 structures 1338:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 8 4 from dominant 1441:1448 arg1 samples 1475:1481 the MYCN-nonamplified samples 1453:1481 the MYCN-nonamplified samples 1453:1481 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 3 5 from signaling 554:562 arg1 important 536:544 important 536:544 important 536:544 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 1 6 theme low-risk 178:185 arg1 neuroblastoma 206:218 low-risk, MYCN-nonamplified neuroblastoma 178:218 low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease 178:257 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 8 7 dep smaller 1396:1402 arg1 nonsialylated 1405:1417 nonsialylated 1405:1417 nonsialylated 1405:1417 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 6 8 theme MYCN-nonamplified 862:878 arg1 SY5Y 880:883 MYCN-nonamplified SY5Y 862:883 MYCN-nonamplified SY5Y 862:883 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 8 9 theme monosaccharide 1289:1302 arg1 count 1304:1308 total monosaccharide count 1283:1308 total monosaccharide count 1283:1308 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 6 10 theme NLF 904:906 arg1 lines 913:917 MYCN-amplified NLF cell lines 889:917 MYCN-amplified NLF cell lines 889:917 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 8 11 theme MYCN-nonamplified 1457:1473 arg1 samples 1475:1481 the MYCN-nonamplified samples 1453:1481 the MYCN-nonamplified samples 1453:1481 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 2 12 theme discerning 331:340 arg1 differences 342:352 discerning differences 331:352 discerning differences in signaling pathways that account for this heterogeneity 331:410 Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. 25730103 9 13 theme neuroblastoma 1554:1566 arg1 diagnosis 1573:1581 accurate neuroblastoma risk diagnosis 1545:1581 accurate neuroblastoma risk diagnosis 1545:1581 These results offer clues for deriving marker candidates for accurate neuroblastoma risk diagnosis. 25730103 3 14 theme N-Linked 413:420 arg1 modification 456:467 a common protein modification 439:467 a common protein modification resulting from the attachment of sugars to protein residues 439:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 14 theme N-Linked 413:420 arg1 glycosylation 422:434 N-Linked glycosylation 413:434 N-Linked glycosylation 413:434 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 6 15 theme N-linked 826:833 arg1 variation 844:852 N-linked glycomic variation 826:852 N-linked glycomic variation 826:852 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 6 16 theme MYCN-amplified 889:902 arg1 lines 913:917 MYCN-amplified NLF cell lines 889:917 MYCN-amplified NLF cell lines 889:917 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 1 17 theme MYCN 64:67 arg1 amplification 69:81 MYCN amplification 64:81 MYCN amplification 64:81 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 9 18 theme risk 1568:1571 arg1 diagnosis 1573:1581 accurate neuroblastoma risk diagnosis 1545:1581 accurate neuroblastoma risk diagnosis 1545:1581 These results offer clues for deriving marker candidates for accurate neuroblastoma risk diagnosis. 25730103 8 19 theme total 1283:1287 arg1 count 1304:1308 total monosaccharide count 1283:1308 total monosaccharide count 1283:1308 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 4 20 link N-linked 594:601 arg1 glycosylation 603:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 0 21 link N-linked 0:7 arg1 glycan 9:14 N-linked glycan 0:14 N-linked glycan 0:14 N-linked glycan profiling in neuroblastoma cell lines. 25730103 6 22 attach linked 974:979 arg1 neuroblastoma 994:1006 high-risk neuroblastoma 984:1006 high-risk neuroblastoma 984:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 6 22 attach linked 974:979 arg2 abundance 964:972 sugar abundance 958:972 sugar abundance linked to high-risk neuroblastoma 958:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 8 23 theme glycan 1331:1336 arg1 structures 1338:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 8 24 theme sialylated 1320:1329 arg1 structures 1338:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 3 25 theme common 441:446 arg1 modification 456:467 a common protein modification 439:467 a common protein modification resulting from the attachment of sugars to protein residues 439:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 25 theme common 441:446 arg1 glycosylation 422:434 N-Linked glycosylation 413:434 N-Linked glycosylation 413:434 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 26 theme sugars 502:507 arg1 attachment 488:497 the attachment 484:497 the attachment of sugars to protein residues 484:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 7 27 theme chromatography-mass 1041:1059 arg1 spectrometry 1061:1072 liquid chromatography-mass spectrometry 1034:1072 liquid chromatography-mass spectrometry 1034:1072 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 6 28 theme sugar 958:962 arg1 abundance 964:972 sugar abundance 958:972 sugar abundance linked to high-risk neuroblastoma 958:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 3 29 from important 536:544 arg1 signaling 554:562 cell signaling 549:562 cell signaling 549:562 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 29 from important 536:544 arg1 response 575:582 immune response 568:582 immune response 568:582 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 2 30 theme signaling 357:365 arg1 pathways 367:374 signaling pathways 357:374 signaling pathways that account for this heterogeneity 357:410 Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. 25730103 0 31 theme N-linked 0:7 arg1 glycan 9:14 N-linked glycan 0:14 N-linked glycan 0:14 N-linked glycan profiling in neuroblastoma cell lines. 25730103 7 32 theme SY5Y 1202:1205 arg1 samples 1207:1213 NLF and SY5Y samples 1194:1213 samples 1207:1213 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 3 33 attach attachment 488:497 arg2 sugars 502:507 sugars 502:507 sugars 502:507 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 33 attach attachment 488:497 arg1 residues 520:527 protein residues 512:527 protein residues 512:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 1 34 dep low-risk 178:185 arg1 MYCN-nonamplified 188:204 MYCN-nonamplified 188:204 MYCN-nonamplified 188:204 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 8 35 theme MYCN-amplified 1356:1369 arg1 samples 1371:1377 the MYCN-amplified samples 1352:1377 the MYCN-amplified samples 1352:1377 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 3 36 from response 575:582 arg1 important 536:544 important 536:544 important 536:544 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 6 37 theme cell 908:911 arg1 lines 913:917 MYCN-amplified NLF cell lines 889:917 MYCN-amplified NLF cell lines 889:917 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 1 38 theme high-risk 225:233 arg1 disease 251:257 high-risk, MYCN-amplified disease 225:257 high-risk, MYCN-amplified disease 225:257 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 6 39 link N-linked 826:833 arg1 variation 844:852 N-linked glycomic variation 826:852 N-linked glycomic variation 826:852 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 6 40 theme variation 844:852 arg1 comparison 812:821 a systematic comparison 799:821 a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma 799:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 7 41 theme NLF 1194:1196 arg1 samples 1207:1213 NLF and SY5Y samples 1194:1213 samples 1207:1213 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 1 42 theme aggressive 108:117 arg1 neuroblastoma 119:131 aggressive neuroblastoma 108:131 aggressive neuroblastoma 108:131 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 7 43 theme spectrometry 1061:1072 arg1 combination 1019:1029 a combination 1017:1029 a combination of liquid chromatography-mass spectrometry and bioinformatics analysis 1017:1100 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 6 44 with lines 913:917 arg1 aim 928:930 the aim 924:930 the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma 924:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 0 45 theme cell 43:46 arg1 lines 48:52 neuroblastoma cell lines 29:52 neuroblastoma cell lines 29:52 N-linked glycan profiling in neuroblastoma cell lines. 25730103 8 46 theme smaller 1396:1402 arg1 glycans 1419:1425 smaller, nonsialylated glycans 1396:1425 smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples 1396:1481 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 0 47 theme neuroblastoma 29:41 arg1 lines 48:52 neuroblastoma cell lines 29:52 neuroblastoma cell lines 29:52 N-linked glycan profiling in neuroblastoma cell lines. 25730103 4 48 theme Aberrant 585:592 arg1 glycosylation 603:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 9 49 theme accurate 1545:1552 arg1 diagnosis 1573:1581 accurate neuroblastoma risk diagnosis 1545:1581 accurate neuroblastoma risk diagnosis 1545:1581 These results offer clues for deriving marker candidates for accurate neuroblastoma risk diagnosis. 25730103 6 50 theme glycomic 835:842 arg1 variation 844:852 N-linked glycomic variation 826:852 N-linked glycomic variation 826:852 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 3 51 theme protein 512:518 arg1 residues 520:527 protein residues 512:527 protein residues 512:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 52 theme protein 448:454 arg1 modification 456:467 a common protein modification 439:467 a common protein modification resulting from the attachment of sugars to protein residues 439:527 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 3 52 theme protein 448:454 arg1 glycosylation 422:434 N-Linked glycosylation 413:434 N-Linked glycosylation 413:434 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 8 53 gly sialylated 1320:1329 arg1 structures 1338:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 larger (in terms of total monosaccharide count) and more sialylated glycan structures 1263:1347 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 7 54 theme significant 1154:1164 arg1 change 1166:1171 a statistically significant change 1138:1171 a statistically significant change in abundance between NLF and SY5Y samples 1138:1213 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 1 55 from disease 251:257 arg1 neuroblastoma 206:218 low-risk, MYCN-nonamplified neuroblastoma 178:218 low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease 178:257 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 6 56 theme high-risk 984:992 arg1 neuroblastoma 994:1006 high-risk neuroblastoma 984:1006 high-risk neuroblastoma 984:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 7 57 theme liquid 1034:1039 arg1 spectrometry 1061:1072 liquid chromatography-mass spectrometry 1034:1072 liquid chromatography-mass spectrometry 1034:1072 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 8 58 theme Closer 1216:1221 arg1 examination 1223:1233 Closer examination 1216:1233 Closer examination 1216:1233 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 8 59 dep larger 1263:1268 arg1 terms 1274:1278 terms 1274:1278 terms of total monosaccharide count 1274:1308 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 3 60 theme cell 549:552 arg1 signaling 554:562 cell signaling 549:562 cell signaling 549:562 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 4 61 theme N-linked 594:601 arg1 glycosylation 603:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 7 62 theme bioinformatics 1078:1091 arg1 analysis 1093:1100 bioinformatics analysis 1078:1100 bioinformatics analysis 1078:1100 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 6 63 from changes 947:953 arg1 abundance 964:972 sugar abundance 958:972 sugar abundance linked to high-risk neuroblastoma 958:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 4 64 attach linked 636:641 arg1 cancers 654:660 various cancers 646:660 various cancers 646:660 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 4 64 attach linked 636:641 arg2 glycosylation 603:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation 585:615 Aberrant N-linked glycosylation has been routinely linked to various cancers. 25730103 2 65 theme proteomic 292:300 arg1 studies 302:308 Genomic and proteomic studies 280:308 Genomic and proteomic studies 280:308 Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. 25730103 6 66 theme systematic 801:810 arg1 comparison 812:821 a systematic comparison 799:821 a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma 799:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 1 67 theme molecular 138:146 arg1 unknown 271:277 unknown 271:277 unknown 271:277 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 1 67 theme molecular 138:146 arg1 mechanisms 148:157 the molecular mechanisms 134:157 the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease 134:257 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 1 68 dep high-risk 225:233 arg1 MYCN-amplified 236:249 MYCN-amplified 236:249 MYCN-amplified 236:249 Although MYCN amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, MYCN-nonamplified neuroblastoma from high-risk, MYCN-amplified disease are largely unknown. 25730103 3 69 theme immune 568:573 arg1 response 575:582 immune response 568:582 immune response 568:582 N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. 25730103 8 70 from samples 1475:1481 arg1 dominant 1441:1448 dominant 1441:1448 dominant 1441:1448 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 9 71 theme marker 1523:1528 arg1 candidates 1530:1539 marker candidates 1523:1539 marker candidates for accurate neuroblastoma risk diagnosis 1523:1581 These results offer clues for deriving marker candidates for accurate neuroblastoma risk diagnosis. 25730103 6 72 with SY5Y 880:883 arg1 aim 928:930 the aim 924:930 the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma 924:1006 Here, we perform a systematic comparison of N-linked glycomic variation between MYCN-nonamplified SY5Y and MYCN-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. 25730103 2 73 theme Genomic 280:286 arg1 studies 302:308 Genomic and proteomic studies 280:308 Genomic and proteomic studies 280:308 Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. 25730103 7 74 theme analysis 1093:1100 arg1 combination 1019:1029 a combination 1017:1029 a combination of liquid chromatography-mass spectrometry and bioinformatics analysis 1017:1100 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 25730103 8 75 theme count 1304:1308 arg1 terms 1274:1278 terms 1274:1278 terms of total monosaccharide count 1274:1308 Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the MYCN-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the MYCN-nonamplified samples. 25730103 2 76 from differences 342:352 arg1 pathways 367:374 signaling pathways 357:374 signaling pathways that account for this heterogeneity 357:410 Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. 25730103 7 77 from change 1166:1171 arg1 abundance 1176:1184 abundance 1176:1184 abundance 1176:1184 Through a combination of liquid chromatography-mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. 28493133 5 0 theme typical 868:874 arg1 methods 876:882 The workflow and typical methods 851:882 The workflow and typical methods involved in the experiment process 851:917 The workflow and typical methods involved in the experiment process will be presented. 28493133 1 1 theme due 238:240 arg1 proteins 229:236 complex recombinant proteins 209:236 complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 209:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 0 2 theme Production 79:88 arg1 N-Glycosylation 40:54 N-Glycosylation 40:54 N-Glycosylation of Recombinant Protein Production in CHO Cell Culture 40:108 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 1 3 theme similar 345:351 arg1 production 289:298 industrial scale production 272:298 industrial scale production 272:298 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 0 4 theme Cell 97:100 arg1 Culture 102:108 CHO Cell Culture 93:108 CHO Cell Culture 93:108 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 3 5 theme genetic 689:695 arg1 elements 697:704 genetic elements 689:704 medium, feed, culture process, as well as genetic elements of the cell 647:716 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 6 theme Protein 560:566 arg1 N-glycosylation 568:582 Protein N-glycosylation 560:582 Protein N-glycosylation in CHO cell culture 560:602 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 7 from N-glycosylation 568:582 arg1 culture 596:602 CHO cell culture 587:602 CHO cell culture 587:602 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 0 8 theme CHO 93:95 arg1 Culture 102:108 CHO Cell Culture 93:108 CHO Cell Culture 93:108 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 1 9 theme long-term 251:259 arg1 success 261:267 their long-term success 245:267 their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 245:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 0 10 from N-Glycosylation 40:54 arg1 Culture 102:108 CHO Cell Culture 93:108 CHO Cell Culture 93:108 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 5 11 theme workflow 855:862 arg1 methods 876:882 The workflow and typical methods 851:882 The workflow and typical methods involved in the experiment process 851:917 The workflow and typical methods involved in the experiment process will be presented. 28493133 2 12 theme physicochemical 508:522 arg1 properties 524:533 both biological and physicochemical properties 488:533 both biological and physicochemical properties of recombinant proteins 488:557 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 3 13 theme cell 713:716 arg1 elements 697:704 genetic elements 689:704 medium, feed, culture process, as well as genetic elements of the cell 647:716 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 13 theme cell 713:716 arg1 feed 655:658 feed 655:658 feed 655:658 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 13 theme cell 713:716 arg1 process 669:675 culture process 661:675 culture process 661:675 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 13 theme cell 713:716 arg1 medium 647:652 medium 647:652 medium 647:652 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 1 14 from success 261:267 arg1 N-glycans 335:343 generating appropriate protein N-glycans 304:343 generating appropriate protein N-glycans 304:343 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 14 from success 261:267 arg1 production 289:298 industrial scale production 272:298 industrial scale production 272:298 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 3 15 theme cell 591:594 arg1 culture 596:602 CHO cell culture 587:602 CHO cell culture 587:602 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 1 16 theme primary 161:167 arg1 system 180:185 the primary expression system 157:185 the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 157:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 17 theme expression 169:178 arg1 system 180:185 the primary expression system 157:185 the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 157:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 4 18 dep medium 821:826 arg1 optimization 837:848 optimization 837:848 optimization 837:848 In this chapter, we will focus on how to carry out experiments for N-glycosylation modulation through medium and feed optimization. 28493133 1 19 theme industrial 272:281 arg1 production 289:298 industrial scale production 272:298 industrial scale production 272:298 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 3 20 theme CHO 587:589 arg1 culture 596:602 CHO cell culture 587:602 CHO cell culture 587:602 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 21 gly N-glycosylation 568:582 arg1 culture 596:602 CHO cell culture 587:602 CHO cell culture 587:602 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 1 22 theme scale 283:287 arg1 production 289:298 industrial scale production 272:298 industrial scale production 272:298 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 3 23 theme culture 661:667 arg1 process 669:675 culture process 661:675 culture process 661:675 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 3 23 theme culture 661:667 arg1 medium 647:652 medium 647:652 medium 647:652 Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell. 28493133 4 24 theme N-glycosylation 786:800 arg1 modulation 802:811 N-glycosylation modulation 786:811 N-glycosylation modulation 786:811 In this chapter, we will focus on how to carry out experiments for N-glycosylation modulation through medium and feed optimization. 28493133 2 25 theme N-glycosylation 408:422 arg1 optimization 384:395 optimization 384:395 optimization 384:395 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 2 25 theme N-glycosylation 408:422 arg1 Control 372:378 Control 372:378 Control 372:378 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 2 26 theme proteins 550:557 arg1 properties 524:533 both biological and physicochemical properties 488:533 both biological and physicochemical properties of recombinant proteins 488:557 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 5 27 theme experiment 900:909 arg1 process 911:917 the experiment process 896:917 the experiment process 896:917 The workflow and typical methods involved in the experiment process will be presented. 28493133 2 28 theme protein 400:406 arg1 N-glycosylation 408:422 protein N-glycosylation 400:422 protein N-glycosylation 400:422 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 2 29 theme N-glycans 456:464 arg1 structure 443:451 the structure 439:451 the structure of N-glycans 439:464 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 2 30 theme recombinant 538:548 arg1 proteins 550:557 recombinant proteins 538:557 recombinant proteins 538:557 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 2 31 theme biological 493:502 arg1 properties 524:533 both biological and physicochemical properties 488:533 both biological and physicochemical properties of recombinant proteins 488:557 Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. 28493133 1 32 theme Chinese 111:117 arg1 cells 139:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 33 theme generating 304:313 arg1 N-glycans 335:343 generating appropriate protein N-glycans 304:343 generating appropriate protein N-glycans 304:343 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 34 theme hamster 119:125 arg1 cells 139:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 35 theme appropriate 315:325 arg1 N-glycans 335:343 generating appropriate protein N-glycans 304:343 generating appropriate protein N-glycans 304:343 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 36 theme ovary 127:131 arg1 cells 139:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 37 theme complex 209:215 arg1 proteins 229:236 complex recombinant proteins 209:236 complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 209:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 0 38 theme Protein 71:77 arg1 Production 79:88 Recombinant Protein Production 59:88 Recombinant Protein Production 59:88 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 0 39 gly N-Glycosylation 40:54 arg1 Production 79:88 Recombinant Protein Production 59:88 Recombinant Protein Production 59:88 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 1 40 theme CHO 134:136 arg1 cells 139:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells 111:143 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 41 theme recombinant 217:227 arg1 proteins 229:236 complex recombinant proteins 209:236 complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 209:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 0 42 theme Recombinant 59:69 arg1 Production 79:88 Recombinant Protein Production 59:88 Recombinant Protein Production 59:88 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture. 28493133 1 43 theme protein 327:333 arg1 N-glycans 335:343 generating appropriate protein N-glycans 304:343 generating appropriate protein N-glycans 304:343 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28493133 1 44 theme proteins 229:236 arg1 production 195:204 the production 191:204 the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans 191:369 Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. 28988842 5 0 gly glycoproteins 938:950 arg1 glycoproteins 938:950 glycoproteins 938:950 glycoproteins containing mucin-type O-glycans 938:982 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 2 1 theme novel 311:315 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 1 theme novel 311:315 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 6 2 theme core 1091:1094 arg1 glycans 1107:1113 core 1 O-linked glycans 1091:1113 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 3 3 theme chromatography 640:653 arg1 sequence 571:578 a sequence 569:578 a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography 569:653 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 1 4 theme carbohydrate 241:252 arg1 biomarkers 254:263 specific carbohydrate biomarkers 232:263 specific carbohydrate biomarkers of many diseases 232:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 4 theme carbohydrate 241:252 arg1 O-glycans 112:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 2 5 link O-linked 324:331 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 5 link O-linked 324:331 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 1 6 theme cell 166:169 arg1 surface 171:177 the cell surface 162:177 the cell surface 162:177 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 4 7 theme apparent 772:779 arg1 protein 791:797 an apparent hexameric protein 769:797 an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3 769:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 7 theme apparent 772:779 arg1 HeL 762:764 The resulting purified HeL 739:764 The resulting purified HeL 739:764 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 7 8 theme extended 1312:1319 arg1 structures 1337:1346 extended core 1 O-glycan structures 1312:1346 extended core 1 O-glycan structures 1312:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 2 9 theme sialic 491:496 arg1 lectin 511:516 a sialic acid-binding lectin 489:516 a sialic acid-binding lectin 489:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 3 10 theme highest 671:677 arg1 yield 679:683 the highest yield 667:683 the highest yield 667:683 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 0 11 link O-linked 15:22 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 7 12 theme range 1303:1307 arg1 detection 1280:1288 detection 1280:1288 detection of a broader range of extended core 1 O-glycan structures 1280:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 9 13 theme glycan 1596:1601 arg1 structures 1603:1612 O- glycan structures 1593:1612 O- glycan structures 1593:1612 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 6 14 theme O-glycan 1135:1142 arg1 structures 1144:1153 extended O-glycan structures 1126:1153 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 6 15 link O-linked 1098:1105 arg1 glycans 1107:1113 core 1 O-linked glycans 1091:1113 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 4 16 theme molecular 814:822 arg1 weight 824:829 a subunit molecular weight 804:829 a subunit molecular weight of 15kDa 804:838 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 3 17 theme protease 720:727 arg1 activity 729:736 protease activity 720:736 protease activity 720:736 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 5 18 theme mucin-type 963:972 arg1 O-glycans 974:982 mucin-type O-glycans 963:982 mucin-type O-glycans 963:982 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 7 19 theme structures 1337:1346 arg1 range 1303:1307 a broader range 1293:1307 a broader range of extended core 1 O-glycan structures 1293:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 4 20 with protein 791:797 arg1 weight 824:829 a subunit molecular weight 804:829 a subunit molecular weight of 15kDa 804:838 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 20 with protein 791:797 arg1 pI 847:848 a pI 845:848 a pI of 4.3 845:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 2 21 theme erinaceus 437:445 arg1 body 407:410 the fruiting body 394:410 the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin 394:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 6 22 theme O-linked 1098:1105 arg1 glycans 1107:1113 core 1 O-linked glycans 1091:1113 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 3 23 theme lectin 705:710 arg1 yield 679:683 the highest yield 667:683 the highest yield 667:683 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 3 23 theme lectin 705:710 arg1 quality 694:700 best quality 689:700 best quality of lectin without protease activity 689:736 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 1 24 theme biological 138:147 arg1 functions 149:157 biological functions 138:157 biological functions on the cell surface as well as the glycoproteins 138:206 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 9 25 theme O- 1593:1594 arg1 structures 1603:1612 O- glycan structures 1593:1612 O- glycan structures 1593:1612 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 2 26 theme mushroom 419:426 arg1 source 478:483 the natural source 466:483 the natural source for a sialic acid-binding lectin 466:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 26 theme mushroom 419:426 arg1 erinaceus 437:445 the mushroom Hericium erinaceus 415:445 the mushroom Hericium erinaceus 415:445 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 0 27 theme fruiting 64:71 arg1 body 73:76 the fruiting body 60:76 the fruiting body of Hericium erinaceus 60:98 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 8 28 theme efficient 1390:1398 arg1 procedure 1414:1422 an efficient and optimized procedure 1387:1422 an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus 1387:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 1 29 from functions 149:157 arg1 surface 171:177 the cell surface 162:177 the cell surface 162:177 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 29 from functions 149:157 arg1 glycoproteins 194:206 the glycoproteins 190:206 the glycoproteins 190:206 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 3 30 theme conditions 557:566 arg1 optimization 524:535 optimization 524:535 optimization of the purification conditions 524:566 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 5 31 theme Galβ 1008:1011 arg1 GalNAc 1017:1022 Galβ(1,3)GalNAc 1008:1022 Galβ(1,3)GalNAc 1008:1022 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 5 32 theme hemagglutination 861:876 arg1 assay 889:893 hemagglutination inhibition assay 861:893 hemagglutination inhibition assay 861:893 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 0 33 theme erinaceus 90:98 arg1 body 73:76 the fruiting body 60:76 the fruiting body of Hericium erinaceus 60:98 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 5 34 theme 1,3 1013:1015 arg1 GalNAc 1017:1022 Galβ(1,3)GalNAc 1008:1022 Galβ(1,3)GalNAc 1008:1022 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 9 35 theme extended 1577:1584 arg1 core 1586:1589 extended core 1 1577:1591 extended core 1 1577:1591 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 2 36 theme specific 340:347 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 36 theme specific 340:347 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 9 37 dep core 1566:1569 arg1 structures 1603:1612 O- glycan structures 1593:1612 O- glycan structures 1593:1612 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 0 38 theme core 8:11 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 1 39 theme many 268:271 arg1 diseases 273:280 many diseases 268:280 many diseases 268:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 7 40 theme HeL 1229:1231 arg1 comparable 1236:1245 comparable 1236:1245 comparable 1236:1245 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 7 40 theme HeL 1229:1231 arg1 specificity 1214:1224 The glycan binding specificity 1195:1224 The glycan binding specificity of HeL 1195:1231 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 8 41 theme fruiting 1461:1468 arg1 body 1470:1473 the fruiting body 1457:1473 the fruiting body of the mushroom Hericium erinaceus 1457:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 2 42 theme erinaceus 366:374 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 42 theme erinaceus 366:374 arg1 HeL 383:385 HeL 383:385 HeL 383:385 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 42 theme erinaceus 366:374 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 0 43 theme O-linked 15:22 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 5 44 contain containing 952:961 arg1 glycoproteins 938:950 glycoproteins 938:950 glycoproteins containing mucin-type O-glycans 938:982 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 5 44 contain containing 952:961 arg2 O-glycans 974:982 mucin-type O-glycans 963:982 mucin-type O-glycans 963:982 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 4 45 theme purified 753:760 arg1 protein 791:797 an apparent hexameric protein 769:797 an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3 769:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 45 theme purified 753:760 arg1 HeL 762:764 The resulting purified HeL 739:764 The resulting purified HeL 739:764 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 0 46 theme binding 40:46 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 7 47 theme glycan 1199:1204 arg1 comparable 1236:1245 comparable 1236:1245 comparable 1236:1245 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 7 47 theme glycan 1199:1204 arg1 specificity 1214:1224 The glycan binding specificity 1195:1224 The glycan binding specificity of HeL 1195:1231 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 8 48 theme mushroom 1482:1489 arg1 erinaceus 1500:1508 the mushroom Hericium erinaceus 1478:1508 the mushroom Hericium erinaceus 1478:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 3 49 theme affinity 597:604 arg1 sequence 571:578 a sequence 569:578 a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography 569:653 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 9 50 theme diagnosis 1617:1625 arg1 assays 1627:1632 diagnosis assays 1617:1632 diagnosis assays 1617:1632 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 8 51 theme erinaceus 1500:1508 arg1 body 1470:1473 the fruiting body 1457:1473 the fruiting body of the mushroom Hericium erinaceus 1457:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 3 52 theme exchange 587:594 arg1 sequence 571:578 a sequence 569:578 a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography 569:653 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 2 53 theme O-linked 324:331 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 53 theme O-linked 324:331 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 54 theme fruiting 398:405 arg1 body 407:410 the fruiting body 394:410 the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin 394:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 9 55 theme powerful 1538:1545 arg1 tool 1547:1550 a powerful tool 1536:1550 a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays 1536:1632 Moreover, HeL represents a powerful tool for analyzing core 1 and extended core 1 O- glycan structures in diagnosis assays. 28988842 8 56 from body 1470:1473 arg1 purification 1432:1443 the purification 1428:1443 the purification of HeL from the fruiting body of the mushroom Hericium erinaceus 1428:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 2 57 theme core 317:320 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 57 theme core 317:320 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 4 58 theme hexameric 781:789 arg1 protein 791:797 an apparent hexameric protein 769:797 an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3 769:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 58 theme hexameric 781:789 arg1 HeL 762:764 The resulting purified HeL 739:764 The resulting purified HeL 739:764 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 3 59 theme ion 607:609 arg1 exchange 611:618 ion exchange 607:618 ion exchange 607:618 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 1 60 theme specific 232:239 arg1 biomarkers 254:263 specific carbohydrate biomarkers 232:263 specific carbohydrate biomarkers of many diseases 232:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 60 theme specific 232:239 arg1 O-glycans 112:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 7 61 theme O-glycan 1328:1335 arg1 structures 1337:1346 extended core 1 O-glycan structures 1312:1346 extended core 1 O-glycan structures 1312:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 3 62 theme size-exclusion 625:638 arg1 chromatography 640:653 size-exclusion chromatography 625:653 size-exclusion chromatography 625:653 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 1 63 gly glycoproteins 194:206 arg1 glycoproteins 194:206 the glycoproteins 190:206 the glycoproteins 190:206 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 7 64 theme core 1321:1324 arg1 structures 1337:1346 extended core 1 O-glycan structures 1312:1346 extended core 1 O-glycan structures 1312:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 4 65 theme 15kDa 834:838 arg1 weight 824:829 a subunit molecular weight 804:829 a subunit molecular weight of 15kDa 804:838 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 65 theme 15kDa 834:838 arg1 pI 847:848 a pI 845:848 a pI of 4.3 845:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 0 66 from body 73:76 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 6 67 theme Glycan 1025:1030 arg1 analyses 1038:1045 Glycan array analyses 1025:1045 Glycan array analyses 1025:1045 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 6 68 theme extended 1126:1133 arg1 structures 1144:1153 extended O-glycan structures 1126:1153 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 1 69 theme Mucin-type 101:110 arg1 biomarkers 254:263 specific carbohydrate biomarkers 232:263 specific carbohydrate biomarkers of many diseases 232:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 69 theme Mucin-type 101:110 arg1 O-glycans 112:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 4 70 theme subunit 806:812 arg1 weight 824:829 a subunit molecular weight 804:829 a subunit molecular weight of 15kDa 804:838 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 3 71 theme best 689:692 arg1 quality 694:700 best quality 689:700 best quality of lectin without protease activity 689:736 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 3 72 theme purification 544:555 arg1 conditions 557:566 the purification conditions 540:566 the purification conditions 540:566 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 2 73 theme acid-binding 498:509 arg1 lectin 511:516 a sialic acid-binding lectin 489:516 a sialic acid-binding lectin 489:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 74 theme Hericium 428:435 arg1 source 478:483 the natural source 466:483 the natural source for a sialic acid-binding lectin 466:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 74 theme Hericium 428:435 arg1 erinaceus 437:445 the mushroom Hericium erinaceus 415:445 the mushroom Hericium erinaceus 415:445 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 8 75 theme optimized 1404:1412 arg1 procedure 1414:1422 an efficient and optimized procedure 1387:1422 an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus 1387:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 7 76 theme peanut 1258:1263 arg1 agglutinin 1265:1274 peanut agglutinin 1258:1274 peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures 1258:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 2 77 theme natural 470:476 arg1 source 478:483 the natural source 466:483 the natural source for a sialic acid-binding lectin 466:516 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 77 theme natural 470:476 arg1 erinaceus 437:445 the mushroom Hericium erinaceus 415:445 the mushroom Hericium erinaceus 415:445 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 0 78 theme Hericium 81:88 arg1 erinaceus 90:98 Hericium erinaceus 81:98 Hericium erinaceus 81:98 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 5 79 theme inhibition 878:887 arg1 assay 889:893 hemagglutination inhibition assay 861:893 hemagglutination inhibition assay 861:893 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 4 80 theme 4.3 853:855 arg1 weight 824:829 a subunit molecular weight 804:829 a subunit molecular weight of 15kDa 804:838 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 80 theme 4.3 853:855 arg1 pI 847:848 a pI 845:848 a pI of 4.3 845:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 8 81 theme HeL 1448:1450 arg1 purification 1432:1443 the purification 1428:1443 the purification of HeL from the fruiting body of the mushroom Hericium erinaceus 1428:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 0 82 theme novel 2:6 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 2 83 theme glycan 333:338 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 83 theme glycan 333:338 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 6 84 contain containing 1155:1164 arg1 structures 1144:1153 extended O-glycan structures 1126:1153 core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation 1091:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 6 84 contain containing 1155:1164 arg2 fucosylation 1181:1192 fucosylation 1181:1192 fucosylation 1181:1192 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 6 84 contain containing 1155:1164 arg2 sialylation 1166:1176 sialylation 1166:1176 sialylation 1166:1176 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 0 85 theme glycan-specific 24:38 arg1 lectin 48:53 A novel core 1 O-linked glycan-specific binding lectin 0:53 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 0:99 A novel core 1 O-linked glycan-specific binding lectin from the fruiting body of Hericium erinaceus. 28988842 1 86 theme diseases 273:280 arg1 biomarkers 254:263 specific carbohydrate biomarkers 232:263 specific carbohydrate biomarkers of many diseases 232:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 86 theme diseases 273:280 arg1 O-glycans 112:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 7 87 theme binding 1206:1212 arg1 comparable 1236:1245 comparable 1236:1245 comparable 1236:1245 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 7 87 theme binding 1206:1212 arg1 specificity 1214:1224 The glycan binding specificity 1195:1224 The glycan binding specificity of HeL 1195:1231 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 2 88 theme Hericium 357:364 arg1 lectin 349:354 a novel core 1 O-linked glycan specific lectin 309:354 a novel core 1 O-linked glycan specific lectin 309:354 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 88 theme Hericium 357:364 arg1 HeL 383:385 HeL 383:385 HeL 383:385 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 2 88 theme Hericium 357:364 arg1 lecin 376:380 Hericium erinaceus lecin 357:380 Hericium erinaceus lecin (HeL) 357:386 In this study, I purified a novel core 1 O-linked glycan specific lectin, Hericium erinaceus lecin (HeL), from the fruiting body of the mushroom Hericium erinaceus, which is known as the natural source for a sialic acid-binding lectin. 28988842 1 89 used used 224:227 arg2 biomarkers 254:263 specific carbohydrate biomarkers 232:263 specific carbohydrate biomarkers of many diseases 232:280 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 1 89 used used 224:227 arg2 O-glycans 112:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans 101:120 Mucin-type O-glycans are involved in biological functions on the cell surface as well as the glycoproteins and can also be used as specific carbohydrate biomarkers of many diseases. 28988842 6 90 theme array 1032:1036 arg1 analyses 1038:1045 Glycan array analyses 1025:1045 Glycan array analyses 1025:1045 Glycan array analyses showed that HeL specifically interacts with core 1 O-linked glycans as well as extended O-glycan structures containing sialylation or fucosylation. 28988842 8 91 theme Hericium 1491:1498 arg1 erinaceus 1500:1508 the mushroom Hericium erinaceus 1478:1508 the mushroom Hericium erinaceus 1478:1508 Taken together, these results provide an efficient and optimized procedure for the purification of HeL from the fruiting body of the mushroom Hericium erinaceus. 28988842 4 92 theme resulting 743:751 arg1 protein 791:797 an apparent hexameric protein 769:797 an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3 769:855 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 4 92 theme resulting 743:751 arg1 HeL 762:764 The resulting purified HeL 739:764 The resulting purified HeL 739:764 The resulting purified HeL is an apparent hexameric protein with a subunit molecular weight of 15kDa, and a pI of 4.3. 28988842 7 93 theme broader 1295:1301 arg1 range 1303:1307 a broader range 1293:1307 a broader range of extended core 1 O-glycan structures 1293:1346 The glycan binding specificity of HeL is comparable to that of peanut agglutinin for detection of a broader range of extended core 1 O-glycan structures. 28988842 3 94 theme ion 583:585 arg1 exchange 587:594 ion exchange 583:594 ion exchange 583:594 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28988842 5 95 theme purified 900:907 arg1 lectin 909:914 the purified lectin 896:914 the purified lectin 896:914 In hemagglutination inhibition assay, the purified lectin was only inhibited by glycoproteins containing mucin-type O-glycans and reacted weakly with Galβ(1,3)GalNAc. 28988842 3 96 theme exchange 611:618 arg1 sequence 571:578 a sequence 569:578 a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography 569:653 Upon optimization of the purification conditions, a sequence of ion exchange, affinity, ion exchange, and size-exclusion chromatography resulted in the highest yield and best quality of lectin without protease activity. 28193332 4 0 theme N-glycosylation 829:843 arg1 mutants 850:856 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 3 1 theme osmium 676:681 arg1 polymer 689:695 an osmium redox polymer 673:695 an osmium redox polymer acting as a redox mediator, up to 10-fold 673:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 2 theme sites 512:516 arg1 mutants 481:487 knock-out mutants 471:487 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 2 theme sites 512:516 arg1 AmPDH1 453:458 In-vitro deglycosylated AmPDH1 429:458 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 4 3 theme mutants 850:856 arg1 set 822:824 a systematic set 809:824 a systematic set of N-glycosylation site mutants 809:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 4 3 theme mutants 850:856 arg1 mutants 850:856 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 7 4 theme polymer 1452:1458 arg1 performance 1402:1412 the active site improved performance 1377:1412 the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1377:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 3 5 theme redox 709:713 arg1 mediator 715:722 a redox mediator 707:722 a redox mediator 707:722 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 6 theme N-glycosylation 496:510 arg1 sites 512:516 the N-glycosylation sites N75 and N175 492:529 the N-glycosylation sites N75 and N175 492:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 6 theme N-glycosylation 496:510 arg1 N175 526:529 N175 526:529 N175 526:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 6 theme N-glycosylation 496:510 arg1 N75 518:520 N75 518:520 N75 518:520 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 6 7 theme structures 1159:1168 arg1 attachment 1113:1122 the attachment 1109:1122 the attachment of significantly extended N-glycan structures 1109:1168 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 6 8 theme enzyme 1253:1258 arg1 performance 1260:1270 enzyme performance 1253:1270 enzyme performance on modified electrodes 1253:1293 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 4 9 gly N-glycosylation 829:843 arg2 mutants 850:856 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 4 9 gly N-glycosylation 829:843 arg2 site 845:848 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 6 10 theme extended 1141:1148 arg1 structures 1159:1168 significantly extended N-glycan structures 1127:1168 significantly extended N-glycan structures 1127:1168 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 7 11 theme improved 1393:1400 arg1 performance 1402:1412 the active site improved performance 1377:1412 the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1377:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 5 12 theme recombinant 1042:1052 arg1 AmPDH1 1064:1069 native and recombinant wild type AmPDH1 1031:1069 native and recombinant wild type AmPDH1 1031:1069 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 3 13 theme site 548:551 arg1 entrance 553:560 the active site entrance 537:560 the active site entrance 537:560 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 1 14 theme capable 228:234 arg1 Dehydrogenase 140:152 Pyranose Dehydrogenase 1 131:154 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) 131:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 14 theme capable 228:234 arg1 oxidoreductase 213:226 an oxidoreductase 210:226 an oxidoreductase capable of oxidizing a broad variety of sugars 210:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 4 15 theme expression 899:908 arg1 efficiency 910:919 expression efficiency 899:919 expression efficiency 899:919 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 7 16 theme active 1381:1386 arg1 performance 1402:1412 the active site improved performance 1377:1412 the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1377:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 5 17 theme type 1059:1062 arg1 AmPDH1 1064:1069 native and recombinant wild type AmPDH1 1031:1069 native and recombinant wild type AmPDH1 1031:1069 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 8 18 theme N-glycosylation 1490:1504 arg1 site 1506:1509 A fourth N-glycosylation site 1481:1509 A fourth N-glycosylation site of AmPDH1 1481:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 18 theme N-glycosylation 1490:1504 arg1 AmPDH1 1514:1519 AmPDH1 1514:1519 AmPDH1 1514:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 5 19 theme native 1031:1036 arg1 AmPDH1 1064:1069 native and recombinant wild type AmPDH1 1031:1069 native and recombinant wild type AmPDH1 1031:1069 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 7 20 theme other 1341:1345 arg1 factors 1347:1353 other factors 1341:1353 other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1341:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 3 21 theme achievable 596:605 arg1 densities 615:623 achievable current densities 596:623 achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold 596:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 1 22 from AmPDH1 199:204 arg1 Dehydrogenase 140:152 Pyranose Dehydrogenase 1 131:154 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) 131:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 22 from AmPDH1 199:204 arg1 oxidoreductase 213:226 an oxidoreductase 210:226 an oxidoreductase capable of oxidizing a broad variety of sugars 210:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 23 theme Agaricus 179:186 arg1 AmPDH1 199:204 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 9 24 theme functional 1795:1804 arg1 expression 1818:1827 functional recombinant expression 1795:1827 functional recombinant expression of the enzyme 1795:1841 This site was shown to be the only one that is essential for functional recombinant expression of the enzyme. 28193332 0 25 theme non-glycosylated 105:120 arg1 enzymes 122:128 partially non-glycosylated enzymes 95:128 partially non-glycosylated enzymes 95:128 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 8 26 theme mass 1543:1546 arg1 spectrometry 1548:1559 mass spectrometry 1543:1559 mass spectrometry 1543:1559 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 4 27 theme better 746:751 arg1 understanding 753:765 a better understanding 744:765 a better understanding of the role of N-glycosylation of AmPDH1 744:806 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 3 28 theme In-vitro 429:436 arg1 AmPDH1 453:458 In-vitro deglycosylated AmPDH1 429:458 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 4 29 gly N-glycosylation 782:796 arg1 AmPDH1 801:806 AmPDH1 801:806 AmPDH1 801:806 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 3 30 dep sites 512:516 arg1 sites 512:516 the N-glycosylation sites N75 and N175 492:529 the N-glycosylation sites N75 and N175 492:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 30 dep sites 512:516 arg1 N175 526:529 N175 526:529 N175 526:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 30 dep sites 512:516 arg1 N75 518:520 N75 518:520 N75 518:520 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 7 31 theme molecule 1323:1330 arg1 size 1332:1335 not the molecule size 1315:1335 not the molecule size 1315:1335 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 6 32 theme polyacrylamide 1185:1198 arg1 electrophoresis 1204:1218 polyacrylamide gel electrophoresis 1185:1218 polyacrylamide gel electrophoresis 1185:1218 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 2 33 theme bio-fuel 413:420 arg1 cells 422:426 enzymatic bio-fuel cells 403:426 enzymatic bio-fuel cells 403:426 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 0 34 theme Agaricus 12:19 arg1 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 7 35 theme AmPDH1/osmium 1432:1444 arg1 polymer 1452:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 1 36 theme Pyranose 131:138 arg1 Dehydrogenase 140:152 Pyranose Dehydrogenase 1 131:154 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) 131:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 36 theme Pyranose 131:138 arg1 oxidoreductase 213:226 an oxidoreductase 210:226 an oxidoreductase capable of oxidizing a broad variety of sugars 210:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 4 37 theme N-glycosylation 782:796 arg1 role 774:777 the role 770:777 the role of N-glycosylation of AmPDH1 770:806 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 0 38 theme pyranose 31:38 arg1 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 3 39 theme graphite 628:635 arg1 electrodes 637:646 graphite electrodes 628:646 graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold 628:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 4 40 theme AmPDH1 801:806 arg1 N-glycosylation 782:796 N-glycosylation 782:796 N-glycosylation of AmPDH1 782:806 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 2 41 theme production 345:354 arg1 ability 296:302 its ability 292:302 its ability of dioxidation of substrates and no side production of hydrogen peroxide 292:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 0 42 theme N-glycosylation 54:68 arg1 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 3 43 gly deglycosylated 438:451 arg1 AmPDH1 453:458 In-vitro deglycosylated AmPDH1 429:458 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 44 theme knock-out 471:479 arg1 mutants 481:487 knock-out mutants 471:487 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 6 45 theme N-glycan 1150:1157 arg1 structures 1159:1168 significantly extended N-glycan structures 1127:1168 significantly extended N-glycan structures 1127:1168 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 6 46 theme site 1089:1092 arg1 N252 1094:1097 the site N252 1085:1097 the site N252 1085:1097 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 8 47 theme fungal 1612:1617 arg1 dehydrogenases 1628:1641 related fungal pyranose dehydrogenases 1604:1641 related fungal pyranose dehydrogenases 1604:1641 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 4 48 theme role 774:777 arg1 understanding 753:765 a better understanding 744:765 a better understanding of the role of N-glycosylation of AmPDH1 744:806 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 8 49 theme glucose-methanol-choline 1675:1698 arg1 family 1726:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 2 50 theme hydrogen 359:366 arg1 peroxide 368:375 hydrogen peroxide 359:375 hydrogen peroxide 359:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 8 51 theme structural 1715:1724 arg1 family 1726:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 1 52 theme sugars 268:273 arg1 variety 257:263 a broad variety 249:263 a broad variety of sugars 249:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 52 theme sugars 268:273 arg1 sugars 268:273 sugars 268:273 sugars 268:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 3 53 theme redox 683:687 arg1 polymer 689:695 an osmium redox polymer 673:695 an osmium redox polymer acting as a redox mediator, up to 10-fold 673:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 6 54 theme modified 1275:1282 arg1 electrodes 1284:1293 modified electrodes 1275:1293 modified electrodes 1275:1293 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 6 55 from performance 1260:1270 arg1 electrodes 1284:1293 modified electrodes 1275:1293 modified electrodes 1275:1293 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 4 56 theme site 845:848 arg1 mutants 850:856 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 8 57 theme other 1654:1658 arg1 members 1660:1666 other members 1654:1666 other members of the glucose-methanol-choline oxidoreductase structural family 1654:1731 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 7 58 theme redox 1446:1450 arg1 polymer 1452:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 2 59 theme substrates 322:331 arg1 dioxidation 307:317 dioxidation 307:317 dioxidation of substrates 307:331 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 2 59 theme substrates 322:331 arg1 production 345:354 no side production 337:354 no side production of hydrogen peroxide 337:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 4 60 theme systematic 811:820 arg1 set 822:824 a systematic set 809:824 a systematic set of N-glycosylation site mutants 809:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 4 60 theme systematic 811:820 arg1 mutants 850:856 N-glycosylation site mutants 829:856 N-glycosylation site mutants 829:856 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 7 61 theme deglycosylated 1417:1430 arg1 polymer 1452:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 2 62 theme dioxidation 307:317 arg1 ability 296:302 its ability 292:302 its ability of dioxidation of substrates and no side production of hydrogen peroxide 292:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 8 63 gly N-glycosylation 1490:1504 arg2 AmPDH1 1514:1519 AmPDH1 1514:1519 AmPDH1 1514:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 63 gly N-glycosylation 1490:1504 arg2 site 1506:1509 A fourth N-glycosylation site 1481:1509 A fourth N-glycosylation site of AmPDH1 1481:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 63 gly N-glycosylation 1490:1504 arg1 AmPDH1 1514:1519 AmPDH1 1514:1519 AmPDH1 1514:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 7 64 theme performance 1402:1412 arg1 accessibility 1360:1372 accessibility 1360:1372 accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1360:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 5 65 dep specific 975:982 arg1 extend 984:989 extend 984:989 extend 984:989 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 5 66 theme wild 1054:1057 arg1 AmPDH1 1064:1069 native and recombinant wild type AmPDH1 1031:1069 native and recombinant wild type AmPDH1 1031:1069 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 8 67 theme fourth 1483:1488 arg1 site 1506:1509 A fourth N-glycosylation site 1481:1509 A fourth N-glycosylation site of AmPDH1 1481:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 67 theme fourth 1483:1488 arg1 AmPDH1 1514:1519 AmPDH1 1514:1519 AmPDH1 1514:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 7 68 theme site 1388:1391 arg1 performance 1402:1412 the active site improved performance 1377:1412 the active site improved performance of deglycosylated AmPDH1/osmium redox polymer 1377:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 3 69 theme current 607:613 arg1 densities 615:623 achievable current densities 596:623 achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold 596:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 70 theme active 541:546 arg1 entrance 553:560 the active site entrance 537:560 the active site entrance 537:560 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 8 71 theme AmPDH1 1514:1519 arg1 site 1506:1509 A fourth N-glycosylation site 1481:1509 A fourth N-glycosylation site of AmPDH1 1481:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 71 theme AmPDH1 1514:1519 arg1 AmPDH1 1514:1519 AmPDH1 1514:1519 AmPDH1 1514:1519 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 6 72 theme gel 1200:1202 arg1 electrophoresis 1204:1218 polyacrylamide gel electrophoresis 1185:1218 polyacrylamide gel electrophoresis 1185:1218 Knocking out the site N252 prevented the attachment of significantly extended N-glycan structures as detected on polyacrylamide gel electrophoresis, but did not significantly alter enzyme performance on modified electrodes. 28193332 1 73 theme basidiomycete 165:177 arg1 AmPDH1 199:204 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 74 theme meleagris 188:196 arg1 AmPDH1 199:204 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 the basidiomycete Agaricus meleagris (AmPDH1) 161:205 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 0 75 theme enzymes 122:128 arg1 Analysis 0:7 Analysis 0:7 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 0:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 0 75 theme enzymes 122:128 arg1 performance 80:90 performance 80:90 performance of partially non-glycosylated enzymes 80:128 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 3 76 dep 10-fold 731:737 arg1 up 725:726 up 725:726 up 725:726 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 9 77 theme recombinant 1806:1816 arg1 expression 1818:1827 functional recombinant expression 1795:1827 functional recombinant expression of the enzyme 1795:1841 This site was shown to be the only one that is essential for functional recombinant expression of the enzyme. 28193332 4 78 theme enzyme 922:927 arg1 activity 929:936 enzyme activity 922:936 enzyme activity 922:936 For a better understanding of the role of N-glycosylation of AmPDH1, a systematic set of N-glycosylation site mutants was investigated in this work, regarding expression efficiency, enzyme activity and stability. 28193332 3 79 gly N-glycosylation 496:510 arg2 N175 526:529 N175 526:529 N175 526:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 79 gly N-glycosylation 496:510 arg2 sites 512:516 the N-glycosylation sites N75 and N175 492:529 the N-glycosylation sites N75 and N175 492:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 79 gly N-glycosylation 496:510 arg2 N75 518:520 N75 518:520 N75 518:520 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 3 80 theme deglycosylated 438:451 arg1 AmPDH1 453:458 In-vitro deglycosylated AmPDH1 429:458 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175 429:529 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 0 81 gly non-glycosylated 105:120 arg1 enzymes 122:128 partially non-glycosylated enzymes 95:128 partially non-glycosylated enzymes 95:128 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 9 82 theme enzyme 1836:1841 arg1 expression 1818:1827 functional recombinant expression 1795:1827 functional recombinant expression of the enzyme 1795:1841 This site was shown to be the only one that is essential for functional recombinant expression of the enzyme. 28193332 0 83 theme meleagris 21:29 arg1 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 3 84 theme electrodes 637:646 arg1 densities 615:623 achievable current densities 596:623 achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold 596:737 In-vitro deglycosylated AmPDH1 as well as knock-out mutants of the N-glycosylation sites N75 and N175, near the active site entrance, were previously shown to improve achievable current densities of graphite electrodes modified with AmPDH1 and an osmium redox polymer acting as a redox mediator, up to 10-fold. 28193332 2 85 theme enzymatic 403:411 arg1 cells 422:426 enzymatic bio-fuel cells 403:426 enzymatic bio-fuel cells 403:426 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 0 86 theme dehydrogenase 40:52 arg1 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 0 87 theme sites 70:74 arg1 Analysis 0:7 Analysis 0:7 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 0:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 0 87 theme sites 70:74 arg1 performance 80:90 performance 80:90 performance of partially non-glycosylated enzymes 80:128 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 5 88 theme N-glycosylation 994:1008 arg1 specific 975:982 specific 975:982 specific 975:982 Furthermore, the site specific extend of N-glycosylation was compared between native and recombinant wild type AmPDH1. 28193332 8 89 theme related 1604:1610 arg1 dehydrogenases 1628:1641 related fungal pyranose dehydrogenases 1604:1641 related fungal pyranose dehydrogenases 1604:1641 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 2 90 theme side 340:343 arg1 production 345:354 no side production 337:354 no side production of hydrogen peroxide 337:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 8 91 theme pyranose 1619:1626 arg1 dehydrogenases 1628:1641 related fungal pyranose dehydrogenases 1604:1641 related fungal pyranose dehydrogenases 1604:1641 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 8 92 theme oxidoreductase 1700:1713 arg1 family 1726:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 the glucose-methanol-choline oxidoreductase structural family 1671:1731 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 28193332 1 93 theme broad 251:255 arg1 variety 257:263 a broad variety 249:263 a broad variety of sugars 249:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 1 93 theme broad 251:255 arg1 sugars 268:273 sugars 268:273 sugars 268:273 Pyranose Dehydrogenase 1 from the basidiomycete Agaricus meleagris (AmPDH1) is an oxidoreductase capable of oxidizing a broad variety of sugars. 28193332 7 94 gly deglycosylated 1417:1430 arg1 polymer 1452:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 deglycosylated AmPDH1/osmium redox polymer 1417:1458 This suggests that not the molecule size but other factors like accessibility of the active site improved performance of deglycosylated AmPDH1/osmium redox polymer modified electrodes. 28193332 0 95 gly N-glycosylation 54:68 arg2 sites 70:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Agaricus meleagris pyranose dehydrogenase N-glycosylation sites 12:74 Analysis of Agaricus meleagris pyranose dehydrogenase N-glycosylation sites and performance of partially non-glycosylated enzymes. 28193332 2 96 from use 396:398 arg1 cells 422:426 enzymatic bio-fuel cells 403:426 enzymatic bio-fuel cells 403:426 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 2 97 theme peroxide 368:375 arg1 dioxidation 307:317 dioxidation 307:317 dioxidation of substrates 307:331 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 2 97 theme peroxide 368:375 arg1 production 345:354 no side production 337:354 no side production of hydrogen peroxide 337:375 Due to this and its ability of dioxidation of substrates and no side production of hydrogen peroxide, it is studied for use in enzymatic bio-fuel cells. 28193332 8 98 theme family 1726:1731 arg1 members 1660:1666 other members 1654:1666 other members of the glucose-methanol-choline oxidoreductase structural family 1654:1731 A fourth N-glycosylation site of AmPDH1 could be confirmed by mass spectrometry at N319, which appeared to be conserved in related fungal pyranose dehydrogenases but not in other members of the glucose-methanol-choline oxidoreductase structural family. 24460129 2 0 theme large 491:495 arg1 area 505:508 large surface area 491:508 large surface area 491:508 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 4 1 theme mSiO2 858:862 arg1 composites 864:873 the C-graphene@mSiO2 composites 843:873 the C-graphene@mSiO2 composites 843:873 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 5 2 theme enrichment 1071:1080 arg1 ability 1082:1088 high selective enrichment ability 1056:1088 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 1 3 theme structure 268:276 arg1 analysis 278:285 a comprehensive structure analysis 252:285 a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer 252:351 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 0 4 theme silica 100:105 arg1 composites 107:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 3 5 theme N-linked 679:686 arg1 glycans 688:694 25 N-linked glycans 676:694 25 N-linked glycans released from ovalbumin 676:718 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 4 6 theme @ 857:857 arg1 composites 864:873 the C-graphene@mSiO2 composites 843:873 the C-graphene@mSiO2 composites 843:873 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 1 7 theme many 194:197 arg1 diseases 199:206 many diseases 194:206 many diseases 194:206 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 4 8 theme sufficient 912:921 arg1 intensities 928:938 sufficient peak intensities 912:938 sufficient peak intensities 912:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 5 9 theme novel 1097:1101 arg1 composite 1120:1128 the novel C-graphene@mSiO2 composite 1093:1128 the novel C-graphene@mSiO2 composite 1093:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 4 10 with enrichment 827:836 arg1 composites 864:873 the C-graphene@mSiO2 composites 843:873 the C-graphene@mSiO2 composites 843:873 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 5 11 theme glycosylation 1160:1172 arg1 research 1174:1181 glycosylation research 1160:1181 glycosylation research 1160:1181 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 2 12 theme pore 522:525 arg1 size 527:530 uniform pore size 514:530 uniform pore size 514:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 1 13 theme glycan 290:295 arg1 analysis 278:285 a comprehensive structure analysis 252:285 a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer 252:351 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 2 14 theme uniform 514:520 arg1 size 527:530 uniform pore size 514:530 uniform pore size 514:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 5 15 theme high 1056:1059 arg1 ability 1082:1088 high selective enrichment ability 1056:1088 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 4 16 theme peak 923:926 arg1 intensities 928:938 sufficient peak intensities 912:938 sufficient peak intensities 912:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 3 17 attach released 696:703 arg1 ovalbumin 710:718 ovalbumin 710:718 ovalbumin 710:718 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 3 17 attach released 696:703 arg2 glycans 688:694 25 N-linked glycans 676:694 25 N-linked glycans released from ovalbumin 676:718 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 2 18 dep composites 445:454 arg1 denoted 457:463 denoted 457:463 denoted as C-graphene@mSiO2 457:483 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 4 19 link N-linked 879:886 arg1 glycans 888:894 48 N-linked glycans 876:894 48 N-linked glycans (S/N > 10) with sufficient peak intensities 876:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 4 19 link N-linked 879:886 arg1 > 901:901 S/N > 10 897:904 S/N > 10 897:904 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 0 20 theme selective 7:15 arg1 enrichment 17:26 Highly selective enrichment 0:26 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 0:117 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 3 21 theme strong 747:752 arg1 signals 757:763 strong MS signals 747:763 strong MS signals 747:763 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 4 22 theme C-graphene 847:856 arg1 composites 864:873 the C-graphene@mSiO2 composites 843:873 the C-graphene@mSiO2 composites 843:873 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 5 23 theme composite 1120:1128 arg1 method 1038:1043 The facile and low-cost synthesis method 1004:1043 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 5 23 theme composite 1120:1128 arg1 ability 1082:1088 high selective enrichment ability 1056:1088 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 2 24 theme first 376:380 arg1 time 382:385 the first time 372:385 the first time 372:385 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 4 25 theme pristine 982:989 arg1 serum 997:1001 healthy pristine human serum 974:1001 healthy pristine human serum 974:1001 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 3 26 theme MS 754:755 arg1 signals 757:763 strong MS signals 747:763 strong MS signals 747:763 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 5 27 theme low-cost 1019:1026 arg1 method 1038:1043 The facile and low-cost synthesis method 1004:1043 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 4 28 theme healthy 974:980 arg1 serum 997:1001 healthy pristine human serum 974:1001 healthy pristine human serum 974:1001 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 1 29 theme Abnormal 119:126 arg1 glycosylation 136:148 Abnormal protein glycosylation 119:148 Abnormal protein glycosylation 119:148 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 3 30 link N-linked 679:686 arg1 glycans 688:694 25 N-linked glycans 676:694 25 N-linked glycans released from ovalbumin 676:718 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 0 31 link N-linked 31:38 arg1 glycan 40:45 N-linked glycan 31:45 N-linked glycan 31:45 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 1 32 theme protein 128:134 arg1 glycosylation 136:148 Abnormal protein glycosylation 119:148 Abnormal protein glycosylation 119:148 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 4 33 with glycans 888:894 arg1 intensities 928:938 sufficient peak intensities 912:938 sufficient peak intensities 912:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 2 34 theme silica 438:443 arg1 composites 445:454 carbon-functionalized ordered graphene/mesoporous silica composites 388:454 carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size 388:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 4 35 theme serum 997:1001 arg1 nL 968:969 only 400 nL 959:969 only 400 nL of healthy pristine human serum 959:1001 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 4 36 theme S/N 897:899 arg1 glycans 888:894 48 N-linked glycans 876:894 48 N-linked glycans (S/N > 10) with sufficient peak intensities 876:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 4 36 theme S/N 897:899 arg1 > 901:901 S/N > 10 897:904 S/N > 10 897:904 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 5 37 theme synthesis 1028:1036 arg1 method 1038:1043 The facile and low-cost synthesis method 1004:1043 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 5 38 theme selective 1061:1069 arg1 ability 1082:1088 high selective enrichment ability 1056:1088 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 2 39 theme graphene/mesoporous 418:436 arg1 composites 445:454 carbon-functionalized ordered graphene/mesoporous silica composites 388:454 carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size 388:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 0 40 theme glycan 40:45 arg1 enrichment 17:26 Highly selective enrichment 0:26 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 0:117 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 0 41 theme ordered 72:78 arg1 composites 107:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 3 42 theme special 590:596 arg1 interaction 598:608 the special interaction 586:608 the special interaction between the carbon and glycans as well as size-exclusion ability 586:673 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 2 43 theme ordered 410:416 arg1 composites 445:454 carbon-functionalized ordered graphene/mesoporous silica composites 388:454 carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size 388:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 0 44 theme N-linked 31:38 arg1 glycan 40:45 N-linked glycan 31:45 N-linked glycan 31:45 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 3 45 theme signal-to-noise 779:793 arg1 ratio 801:805 increased signal-to-noise (S/N) ratio 769:805 increased signal-to-noise (S/N) ratio 769:805 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 3 46 theme interaction 598:608 arg1 advantage 573:581 advantage 573:581 advantage of the special interaction between the carbon and glycans as well as size-exclusion ability 573:673 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 5 47 theme promising 1141:1149 arg1 tool 1151:1154 a promising tool 1139:1154 a promising tool for glycosylation research 1139:1181 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 2 48 theme carbon-functionalized 388:408 arg1 composites 445:454 carbon-functionalized ordered graphene/mesoporous silica composites 388:454 carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size 388:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 2 49 theme @ 478:478 arg1 mSiO2 479:483 C-graphene@mSiO2 468:483 C-graphene@mSiO2 468:483 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 1 50 theme diseases 328:335 arg1 diagnosis 315:323 diagnosis 315:323 diagnosis 315:323 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 1 50 theme diseases 328:335 arg1 prognosis 301:309 prognosis 301:309 prognosis 301:309 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 24460129 4 51 theme N-linked 879:886 arg1 glycans 888:894 48 N-linked glycans 876:894 48 N-linked glycans (S/N > 10) with sufficient peak intensities 876:938 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 4 51 theme N-linked 879:886 arg1 > 901:901 S/N > 10 897:904 S/N > 10 897:904 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 2 52 theme surface 497:503 arg1 area 505:508 large surface area 491:508 large surface area 491:508 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 2 53 with composites 445:454 arg1 area 505:508 large surface area 491:508 large surface area 491:508 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 2 53 with composites 445:454 arg1 size 527:530 uniform pore size 514:530 uniform pore size 514:530 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 2 54 theme C-graphene 468:477 arg1 mSiO2 479:483 C-graphene@mSiO2 468:483 C-graphene@mSiO2 468:483 In this work, for the first time, carbon-functionalized ordered graphene/mesoporous silica composites (denoted as C-graphene@mSiO2) with large surface area and uniform pore size were designed and synthesized. 24460129 5 55 theme mSiO2 1114:1118 arg1 composite 1120:1128 the novel C-graphene@mSiO2 composite 1093:1128 the novel C-graphene@mSiO2 composite 1093:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 5 56 theme C-graphene 1103:1112 arg1 composite 1120:1128 the novel C-graphene@mSiO2 composite 1093:1128 the novel C-graphene@mSiO2 composite 1093:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 3 57 theme increased 769:777 arg1 ratio 801:805 increased signal-to-noise (S/N) ratio 769:805 increased signal-to-noise (S/N) ratio 769:805 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 5 58 theme facile 1008:1013 arg1 method 1038:1043 The facile and low-cost synthesis method 1004:1043 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite 1004:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 3 59 theme size-exclusion 652:665 arg1 ability 667:673 size-exclusion ability 652:673 the special interaction between the carbon and glycans as well as size-exclusion ability 586:673 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 0 60 theme graphene/mesoporous 80:98 arg1 composites 107:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 3 61 theme ability 667:673 arg1 advantage 573:581 advantage 573:581 advantage of the special interaction between the carbon and glycans as well as size-exclusion ability 573:673 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 3 62 theme S/N 796:798 arg1 ratio 801:805 increased signal-to-noise (S/N) ratio 769:805 increased signal-to-noise (S/N) ratio 769:805 By taking advantage of the special interaction between the carbon and glycans as well as size-exclusion ability, 25 N-linked glycans released from ovalbumin were observed clearly with strong MS signals and increased signal-to-noise (S/N) ratio. 24460129 0 63 theme carbon-functionalized 50:70 arg1 composites 107:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 carbon-functionalized ordered graphene/mesoporous silica composites 50:116 Highly selective enrichment of N-linked glycan by carbon-functionalized ordered graphene/mesoporous silica composites. 24460129 5 64 theme @ 1113:1113 arg1 composite 1120:1128 the novel C-graphene@mSiO2 composite 1093:1128 the novel C-graphene@mSiO2 composite 1093:1128 The facile and low-cost synthesis method as well as high selective enrichment ability of the novel C-graphene@mSiO2 composite makes it a promising tool for glycosylation research. 24460129 4 65 theme human 991:995 arg1 serum 997:1001 healthy pristine human serum 974:1001 healthy pristine human serum 974:1001 In addition, after enrichment with the C-graphene@mSiO2 composites, 48 N-linked glycans (S/N > 10) with sufficient peak intensities were obtained from only 400 nL of healthy pristine human serum. 24460129 1 66 theme comprehensive 254:266 arg1 analysis 278:285 a comprehensive structure analysis 252:285 a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer 252:351 Abnormal protein glycosylation has been demonstrated to be associated with many diseases; therefore, it is very important to conduct a comprehensive structure analysis of glycan for prognosis and diagnosis of diseases, such as cancer. 27501865 8 0 theme morphine 1586:1593 arg1 consumption 1595:1605 morphine consumption 1586:1605 morphine consumption 1586:1605 We did not find any statistically significant association between morphine consumption and plasma N-glycome. 27501865 1 1 theme healing 170:176 arg1 process 178:184 the complex healing process 158:184 the complex healing process occurring after major surgery 158:214 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 6 2 gly sialylated 1195:1204 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 1 3 theme patient 266:272 arg1 recovery 274:281 patient recovery 266:281 patient recovery 266:281 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 7 4 theme structures 1413:1422 arg1 decrease 1397:1404 a concomitant decrease 1383:1404 a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans 1383:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 8 5 theme plasma 1611:1616 arg1 N-glycome 1618:1626 plasma N-glycome 1611:1626 plasma N-glycome 1611:1626 We did not find any statistically significant association between morphine consumption and plasma N-glycome. 27501865 7 6 theme triantennary 1498:1509 arg1 glycans 1511:1517 trisialylated triantennary glycans 1484:1517 trisialylated triantennary glycans 1484:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 5 7 theme bisialylated 926:937 arg1 12 hours 972:979 12 hours 972:979 12 hours 972:979 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 7 theme bisialylated 926:937 arg1 A2G2S[3,6 959:967 A2G2S[3,6]2 959:969 A2G2S[3,6]2 959:969 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 7 theme bisialylated 926:937 arg1 glycan 951:956 bisialylated biantennary glycan 926:956 bisialylated biantennary glycan 926:956 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 8 theme biantennary 939:949 arg1 12 hours 972:979 12 hours 972:979 12 hours 972:979 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 8 theme biantennary 939:949 arg1 A2G2S[3,6 959:967 A2G2S[3,6]2 959:969 A2G2S[3,6]2 959:969 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 8 theme biantennary 939:949 arg1 glycan 951:956 bisialylated biantennary glycan 926:956 bisialylated biantennary glycan 926:956 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 7 9 contain containing 1424:1433 arg2 N-acetylglucosamine 1445:1463 bisecting N-acetylglucosamine 1435:1463 bisecting N-acetylglucosamine 1435:1463 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 7 9 contain containing 1424:1433 arg1 structures 1413:1422 the structures 1409:1422 the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans 1409:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 0 10 theme abdominal 101:109 arg1 surgery 111:117 major abdominal surgery 95:117 major abdominal surgery 95:117 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 0 11 from Changes 0:6 arg1 analgesia 79:87 patient-controlled analgesia 60:87 patient-controlled analgesia 60:87 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 0 11 from Changes 0:6 arg1 composition 44:54 total plasma and serum N-glycome composition 11:54 total plasma and serum N-glycome composition 11:54 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 6 12 theme tetraantennary 1206:1219 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 7 13 dep triantennary 1311:1322 arg1 sialylated 1300:1309 sialylated 1300:1309 sialylated 1300:1309 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 0 14 theme major 95:99 arg1 surgery 111:117 major abdominal surgery 95:117 major abdominal surgery 95:117 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 6 15 gly core-fucosylated 1127:1142 arg1 structures 1156:1165 most core-fucosylated biantennary structures 1122:1165 most core-fucosylated biantennary structures 1122:1165 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 3 16 from effects 531:537 arg1 N-glycome 570:578 the total plasma N-glycome 553:578 the total plasma N-glycome 553:578 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 3 16 from effects 531:537 arg1 association 588:598 the association 584:598 the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery 584:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 3 17 theme total 557:561 arg1 N-glycome 570:578 the total plasma N-glycome 553:578 the total plasma N-glycome 553:578 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 7 18 dep days 1358:1361 arg1 surgery 1369:1375 surgery 1369:1375 surgery 1369:1375 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 5 19 theme glycan 951:956 arg1 increase 914:921 the increase 910:921 the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours 910:979 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 6 20 theme core-fucosylated 1127:1142 arg1 structures 1156:1165 most core-fucosylated biantennary structures 1122:1165 most core-fucosylated biantennary structures 1122:1165 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 3 21 theme surgery 542:548 arg1 effects 531:537 the effects 527:537 the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery 527:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 2 22 theme plasma 290:295 arg1 indicator 319:327 an indicator 316:327 an indicator of inflammation 316:343 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 2 22 theme plasma 290:295 arg1 N-glycome 297:305 Total plasma N-glycome 284:305 Total plasma N-glycome 284:305 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 2 23 theme response 406:413 arg1 marker 415:420 response marker 406:420 an anti-inflammatory therapy response marker 377:420 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 6 24 theme sialylated 1195:1204 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 7 25 gly trisialylated 1484:1496 arg1 glycans 1511:1517 trisialylated triantennary glycans 1484:1517 trisialylated triantennary glycans 1484:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 2 26 theme inflammation 332:343 arg1 indicator 319:327 an indicator 316:327 an indicator of inflammation 316:343 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 2 26 theme inflammation 332:343 arg1 N-glycome 297:305 Total plasma N-glycome 284:305 Total plasma N-glycome 284:305 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 1 27 theme major 202:206 arg1 surgery 208:214 major surgery 202:214 major surgery 202:214 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 2 28 theme essential 460:468 arg1 role 470:473 an essential role 457:473 an essential role 457:473 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 7 29 theme trisialylated 1484:1496 arg1 glycans 1511:1517 trisialylated triantennary glycans 1484:1517 trisialylated triantennary glycans 1484:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 3 30 from association 588:598 arg1 cohorts 658:664 two cohorts 654:664 two cohorts of patients that underwent major abdominal surgery 654:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 2 31 theme anti-inflammatory 380:396 arg1 therapy 398:404 an anti-inflammatory therapy 377:404 an anti-inflammatory therapy response marker 377:420 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 0 32 theme plasma 17:22 arg1 composition 44:54 total plasma and serum N-glycome composition 11:54 total plasma and serum N-glycome composition 11:54 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 6 33 from decrease 1110:1117 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 3 34 theme plasma 563:568 arg1 N-glycome 570:578 the total plasma N-glycome 553:578 the total plasma N-glycome 553:578 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 0 35 theme total 11:15 arg1 composition 44:54 total plasma and serum N-glycome composition 11:54 total plasma and serum N-glycome composition 11:54 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 6 36 from increase 1183:1190 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 3 37 theme patients 669:676 arg1 cohorts 658:664 two cohorts 654:664 two cohorts of patients that underwent major abdominal surgery 654:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 6 38 theme most 1122:1125 arg1 structures 1156:1165 most core-fucosylated biantennary structures 1122:1165 most core-fucosylated biantennary structures 1122:1165 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 2 39 theme protein 429:435 arg1 glycosylation 437:449 protein glycosylation 429:449 protein glycosylation 429:449 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 0 40 theme serum 28:32 arg1 composition 44:54 total plasma and serum N-glycome composition 11:54 total plasma and serum N-glycome composition 11:54 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 4 41 dep day 783:785 arg1 surgery 793:799 surgery 793:799 surgery 793:799 We found that plasma N-glycome undergoes significant changes one day after surgery and intensifies one day later, thus indicating a systemic physiological response. 27501865 3 42 with N-glycome 570:578 arg1 self-administration 605:623 self-administration 605:623 self-administration of postoperative morphine 605:649 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 2 43 theme inflammatory 482:493 arg1 cascade 495:501 the inflammatory cascade 478:501 the inflammatory cascade 478:501 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 7 44 theme bisecting 1435:1443 arg1 N-acetylglucosamine 1445:1463 bisecting N-acetylglucosamine 1435:1463 bisecting N-acetylglucosamine 1435:1463 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 8 45 theme significant 1554:1564 arg1 association 1566:1576 any statistically significant association 1536:1576 any statistically significant association between morphine consumption and plasma N-glycome 1536:1626 We did not find any statistically significant association between morphine consumption and plasma N-glycome. 27501865 3 46 with association 588:598 arg1 self-administration 605:623 self-administration 605:623 self-administration of postoperative morphine 605:649 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 4 47 theme physiological 859:871 arg1 response 873:880 a systemic physiological response 848:880 a systemic physiological response 848:880 We found that plasma N-glycome undergoes significant changes one day after surgery and intensifies one day later, thus indicating a systemic physiological response. 27501865 1 48 theme Systemic 120:127 arg1 inflammation 129:140 Systemic inflammation 120:140 Systemic inflammation 120:140 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 0 49 theme N-glycome 34:42 arg1 composition 44:54 total plasma and serum N-glycome composition 11:54 total plasma and serum N-glycome composition 11:54 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 6 50 theme FA3G3S[3,3,3 1225:1236 arg1 structures 1240:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1195:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 3 51 theme abdominal 699:707 arg1 surgery 709:715 major abdominal surgery 693:715 major abdominal surgery 693:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 4 52 theme systemic 850:857 arg1 response 873:880 a systemic physiological response 848:880 a systemic physiological response 848:880 We found that plasma N-glycome undergoes significant changes one day after surgery and intensifies one day later, thus indicating a systemic physiological response. 27501865 7 53 theme tetraantennary 1328:1341 arg1 structures 1343:1352 sialylated triantennary and tetraantennary structures 1300:1352 structures 1343:1352 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 4 54 theme plasma 732:737 arg1 N-glycome 739:747 plasma N-glycome 732:747 plasma N-glycome 732:747 We found that plasma N-glycome undergoes significant changes one day after surgery and intensifies one day later, thus indicating a systemic physiological response. 27501865 5 55 theme postoperative 1035:1047 arg1 hours 1049:1053 48 postoperative hours 1032:1053 48 postoperative hours 1032:1053 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 2 56 dep therapy 398:404 arg1 marker 415:420 response marker 406:420 an anti-inflammatory therapy response marker 377:420 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 2 57 theme Total 284:288 arg1 indicator 319:327 an indicator 316:327 an indicator of inflammation 316:343 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 2 57 theme Total 284:288 arg1 N-glycome 297:305 Total plasma N-glycome 284:305 Total plasma N-glycome 284:305 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 6 58 theme Most 1056:1059 arg1 changes 1061:1067 Most changes 1056:1067 Most changes 1056:1067 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 2 59 theme major 351:355 arg1 surgery 357:363 major surgery 351:363 major surgery 351:363 Total plasma N-glycome might be an indicator of inflammation after major surgery, as well as an anti-inflammatory therapy response marker, since protein glycosylation plays an essential role in the inflammatory cascade. 27501865 3 60 from N-glycome 570:578 arg1 cohorts 658:664 two cohorts 654:664 two cohorts of patients that underwent major abdominal surgery 654:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 5 61 gly bisialylated 926:937 arg1 12 hours 972:979 12 hours 972:979 12 hours 972:979 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 61 gly bisialylated 926:937 arg1 A2G2S[3,6 959:967 A2G2S[3,6]2 959:969 A2G2S[3,6]2 959:969 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 5 61 gly bisialylated 926:937 arg1 glycan 951:956 bisialylated biantennary glycan 926:956 bisialylated biantennary glycan 926:956 In particular, we observed the increase of bisialylated biantennary glycan, A2G2S[3,6]2, 12 hours after surgery, which progressively increased until 48 postoperative hours. 27501865 7 62 theme structures 1343:1352 arg1 increase 1288:1295 a progressive increase 1274:1295 a progressive increase of sialylated triantennary and tetraantennary structures 1274:1352 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 4 63 theme significant 759:769 arg1 changes 771:777 significant changes 759:777 significant changes 759:777 We found that plasma N-glycome undergoes significant changes one day after surgery and intensifies one day later, thus indicating a systemic physiological response. 27501865 6 64 theme biantennary 1144:1154 arg1 structures 1156:1165 most core-fucosylated biantennary structures 1122:1165 most core-fucosylated biantennary structures 1122:1165 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 1 65 theme surgical 245:252 arg1 outcome 254:260 the surgical outcome 241:260 the surgical outcome 241:260 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 0 66 theme patient-controlled 60:77 arg1 analgesia 79:87 patient-controlled analgesia 60:87 patient-controlled analgesia 60:87 Changes in total plasma and serum N-glycome composition and patient-controlled analgesia after major abdominal surgery. 27501865 3 67 theme major 693:697 arg1 surgery 709:715 major abdominal surgery 693:715 major abdominal surgery 693:715 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 3 68 theme postoperative 628:640 arg1 morphine 642:649 postoperative morphine 628:649 postoperative morphine 628:649 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 6 69 theme structures 1156:1165 arg1 decrease 1110:1117 the decrease 1106:1117 the decrease of most core-fucosylated biantennary structures 1106:1165 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 6 69 theme structures 1156:1165 arg1 increase 1183:1190 the increase 1179:1190 the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures 1179:1249 Most changes occurred 24 hours after surgery with the decrease of most core-fucosylated biantennary structures, as well as the increase in sialylated tetraantennary and FA3G3S[3,3,3]3 structures. 27501865 7 70 theme concomitant 1385:1395 arg1 decrease 1397:1404 a concomitant decrease 1383:1404 a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans 1383:1517 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 3 71 theme morphine 642:649 arg1 self-administration 605:623 self-administration 605:623 self-administration of postoperative morphine 605:649 Therefore, we assessed the effects of surgery on the total plasma N-glycome and the association with self-administration of postoperative morphine in two cohorts of patients that underwent major abdominal surgery. 27501865 7 72 theme triantennary 1311:1322 arg1 increase 1288:1295 a progressive increase 1274:1295 a progressive increase of sialylated triantennary and tetraantennary structures 1274:1352 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27501865 1 73 theme complex 162:168 arg1 process 178:184 the complex healing process 158:184 the complex healing process occurring after major surgery 158:214 Systemic inflammation participates to the complex healing process occurring after major surgery, thus directly affecting the surgical outcome and patient recovery. 27501865 7 74 theme progressive 1276:1286 arg1 increase 1288:1295 a progressive increase 1274:1295 a progressive increase of sialylated triantennary and tetraantennary structures 1274:1352 Moreover, we observed a progressive increase of sialylated triantennary and tetraantennary structures two days after surgery, with a concomitant decrease of the structures containing bisecting N-acetylglucosamine along with bi- and trisialylated triantennary glycans. 27975142 1 0 theme backbone 224:231 arg1 constituent 205:215 constituent 205:215 constituent of the backbone of nucleic acids and of cell wall polysaccharides 205:281 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 2 1 theme following 451:459 arg1 addition 534:541 addition 534:541 addition of branches 534:553 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 1 theme following 451:459 arg1 introduction 573:584 site-specific introduction 559:584 site-specific introduction of substitutions 559:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 1 theme following 451:459 arg1 status 496:501 anomeric status 487:501 anomeric status 487:501 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 1 theme following 451:459 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 1 theme following 451:459 arg1 size 528:531 ring size 523:531 ring size 523:531 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 3 2 theme life 658:661 arg1 alphabet 646:653 the third alphabet 636:653 the third alphabet of life 636:661 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 2 3 theme site-specific 559:571 arg1 introduction 573:584 site-specific introduction 559:584 site-specific introduction of substitutions 559:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 3 theme site-specific 559:571 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 4 with parameters 461:470 arg1 sequence 477:484 sequence 477:484 sequence 477:484 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 1 5 theme activated 139:147 arg1 hydrogen 149:156 activated hydrogen 139:156 activated hydrogen 139:156 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 6 6 theme cellular 1134:1141 arg1 effects 1143:1149 cellular effects 1134:1149 cellular effects 1134:1149 Glycan-based 'words' can thus be 'read,' and their message translated into cellular effects by receptors called lectins. 27975142 1 7 theme nucleic 236:242 arg1 acids 244:248 nucleic acids 236:248 nucleic acids 236:248 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 4 8 from lipids 783:788 arg1 words 736:740 words 736:740 These 'words' 729:741 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 8 from lipids 783:788 arg1 part 747:750 part 747:750 part of the glycans on proteins and lipids 747:788 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 2 9 theme branches 546:553 arg1 introduction 573:584 site-specific introduction 559:584 site-specific introduction of substitutions 559:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 9 theme branches 546:553 arg1 status 496:501 anomeric status 487:501 anomeric status 487:501 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 9 theme branches 546:553 arg1 size 528:531 ring size 523:531 ring size 523:531 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 9 theme branches 546:553 arg1 addition 534:541 addition 534:541 addition of branches 534:553 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 9 theme branches 546:553 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 9 theme branches 546:553 arg1 positions 512:520 linkage positions 504:520 linkage positions 504:520 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 1 10 theme physiological 54:66 arg1 importance 68:77 physiological importance 54:77 physiological importance 54:77 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 1 11 theme acids 244:248 arg1 backbone 224:231 the backbone 220:231 the backbone of nucleic acids 220:248 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 2 12 theme anomeric 487:494 arg1 status 496:501 anomeric status 487:501 anomeric status 487:501 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 12 theme anomeric 487:494 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 7 13 dep sugar 1254:1258 arg1 code 1260:1263 code 1260:1263 code 1260:1263 This journal's special issue covers central aspects of the concept of the sugar code. 27975142 1 14 theme hydrogen 149:156 arg1 source 105:110 source 105:110 source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) 105:197 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 4 15 theme cell 858:861 arg1 features 878:885 cell type-dependent features 858:885 cell type-dependent features 858:885 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 16 from proteins 770:777 arg1 words 736:740 words 736:740 These 'words' 729:741 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 16 from proteins 770:777 arg1 part 747:750 part 747:750 part of the glycans on proteins and lipids 747:788 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 6 17 theme Glycan-based 1059:1070 arg1 words 1073:1077 words 1073:1077 Glycan-based 'words' 1059:1078 Glycan-based 'words' can thus be 'read,' and their message translated into cellular effects by receptors called lectins. 27975142 4 18 theme type-dependent 863:876 arg1 features 878:885 cell type-dependent features 858:885 cell type-dependent features 858:885 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 1 19 contain have 49:52 arg2 importance 68:77 physiological importance 54:77 physiological importance 54:77 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 1 19 contain have 49:52 arg1 Carbohydrates 35:47 Carbohydrates 35:47 Carbohydrates 35:47 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 2 20 theme substitutions 589:601 arg1 introduction 573:584 site-specific introduction 559:584 site-specific introduction of substitutions 559:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 20 theme substitutions 589:601 arg1 status 496:501 anomeric status 487:501 anomeric status 487:501 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 20 theme substitutions 589:601 arg1 size 528:531 ring size 523:531 ring size 523:531 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 20 theme substitutions 589:601 arg1 addition 534:541 addition 534:541 addition of branches 534:553 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 20 theme substitutions 589:601 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 20 theme substitutions 589:601 arg1 positions 512:520 linkage positions 504:520 linkage positions 504:520 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 21 theme unique 404:409 arg1 property 411:418 the unique property 400:418 the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 400:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 5 22 theme points 988:993 arg1 abundance 967:975 an abundance 964:975 an abundance of contact points for intermolecular interactions 964:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 3 23 theme high-density 699:710 arg1 capacity 719:726 high-density coding capacity 699:726 high-density coding capacity 699:726 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 2 24 theme ring 523:526 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 24 theme ring 523:526 arg1 size 528:531 ring size 523:531 ring size 523:531 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 1 25 theme pentosephosphate 173:188 arg1 synthesis 162:170 synthesis 162:170 synthesis (pentosephosphate pathway) 162:197 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 1 25 theme pentosephosphate 173:188 arg1 pathway 190:196 pentosephosphate pathway 173:196 pentosephosphate pathway 173:196 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 1 26 theme cell 257:260 arg1 polysaccharides 267:281 cell wall polysaccharides 257:281 cell wall polysaccharides 257:281 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 2 27 theme variability 327:337 arg1 extent 288:293 The extent 284:293 The extent of compositional and structural variability of their oligomers (glycans) 284:366 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 1 28 theme glycolysis 123:132 arg1 source 105:110 source 105:110 source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) 105:197 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 1 29 theme wall 262:265 arg1 polysaccharides 267:281 cell wall polysaccharides 257:281 cell wall polysaccharides 257:281 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 5 30 theme intermolecular 999:1012 arg1 interactions 1014:1025 intermolecular interactions 999:1025 intermolecular interactions 999:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 5 31 theme oligosaccharides 936:951 arg1 flexibility 921:931 The often limited intramolecular flexibility 888:931 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions 888:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 5 31 theme oligosaccharides 936:951 arg1 ideal 1030:1034 ideal 1030:1034 ideal 1030:1034 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 7 32 theme concept 1239:1245 arg1 aspects 1224:1230 central aspects 1216:1230 central aspects of the concept of the sugar code 1216:1263 This journal's special issue covers central aspects of the concept of the sugar code. 27975142 1 33 theme polysaccharides 267:281 arg1 constituent 205:215 constituent 205:215 constituent of the backbone of nucleic acids and of cell wall polysaccharides 205:281 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 2 34 theme linkage 504:510 arg1 positions 512:520 linkage positions 504:520 linkage positions 504:520 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 4 35 from glycans 759:765 arg1 lipids 783:788 lipids 783:788 lipids 783:788 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 35 from glycans 759:765 arg1 proteins 770:777 proteins 770:777 proteins 770:777 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 2 36 dep parameters 461:470 arg1 introduction 573:584 site-specific introduction 559:584 site-specific introduction of substitutions 559:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 36 dep parameters 461:470 arg1 status 496:501 anomeric status 487:501 anomeric status 487:501 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 36 dep parameters 461:470 arg1 size 528:531 ring size 523:531 ring size 523:531 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 36 dep parameters 461:470 arg1 addition 534:541 addition 534:541 addition of branches 534:553 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 36 dep parameters 461:470 arg1 parameters 461:470 the following parameters 447:470 the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions 447:601 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 2 36 dep parameters 461:470 arg1 positions 512:520 linkage positions 504:520 linkage positions 504:520 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 4 37 theme glycans 759:765 arg1 words 736:740 words 736:740 These 'words' 729:741 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 37 theme glycans 759:765 arg1 part 747:750 part 747:750 part of the glycans on proteins and lipids 747:788 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 5 38 theme binding 1040:1046 arg1 processes 1048:1056 binding processes 1040:1056 binding processes 1040:1056 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 2 39 theme structural 316:325 arg1 variability 327:337 compositional and structural variability 298:337 compositional and structural variability of their oligomers (glycans) 298:366 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 3 40 theme coding 712:717 arg1 capacity 719:726 high-density coding capacity 699:726 high-density coding capacity 699:726 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 3 41 theme alphabet 646:653 arg1 letters 625:631 letters 625:631 letters of the third alphabet of life 625:661 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 7 42 theme special 1195:1201 arg1 issue 1203:1207 This journal's special issue 1180:1207 This journal's special issue 1180:1207 This journal's special issue covers central aspects of the concept of the sugar code. 27975142 3 43 theme capacity 719:726 arg1 words 679:683 words 679:683 'words' (signals) of high-density coding capacity 678:726 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 3 43 theme capacity 719:726 arg1 signals 687:693 signals 687:693 signals 687:693 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 5 44 theme limited 898:904 arg1 flexibility 921:931 The often limited intramolecular flexibility 888:931 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions 888:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 5 44 theme limited 898:904 arg1 ideal 1030:1034 ideal 1030:1034 ideal 1030:1034 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 4 45 contain has 854:856 arg2 features 878:885 cell type-dependent features 858:885 cell type-dependent features 858:885 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 45 contain has 854:856 arg1 glycome 799:805 the glycome 795:805 the glycome represented by these 'words' in their entirety 795:852 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 1 46 theme energy 115:120 arg1 glycolysis 123:132 energy (glycolysis) 115:133 energy (glycolysis) 115:133 Carbohydrates have physiological importance far beyond their roles as source of energy (glycolysis) and activated hydrogen for synthesis (pentosephosphate pathway) or as constituent of the backbone of nucleic acids and of cell wall polysaccharides. 27975142 3 47 dep monosaccharides 608:622 arg1 letters 625:631 letters 625:631 letters of the third alphabet of life 625:661 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 4 48 from part 747:750 arg1 lipids 783:788 lipids 783:788 lipids 783:788 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 4 48 from part 747:750 arg1 proteins 770:777 proteins 770:777 proteins 770:777 These 'words' are part of the glycans on proteins and lipids, and the glycome represented by these 'words' in their entirety has cell type-dependent features. 27975142 5 49 theme contact 980:986 arg1 points 988:993 contact points 980:993 contact points for intermolecular interactions 980:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 5 50 theme intramolecular 906:919 arg1 flexibility 921:931 The often limited intramolecular flexibility 888:931 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions 888:1025 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 5 50 theme intramolecular 906:919 arg1 ideal 1030:1034 ideal 1030:1034 ideal 1030:1034 The often limited intramolecular flexibility of oligosaccharides along with an abundance of contact points for intermolecular interactions is ideal for binding processes. 27975142 2 51 theme compositional 298:310 arg1 variability 327:337 compositional and structural variability 298:337 compositional and structural variability of their oligomers (glycans) 298:366 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 3 52 theme third 640:644 arg1 alphabet 646:653 the third alphabet 636:653 the third alphabet of life 636:661 The monosaccharides (letters of the third alphabet of life) thus generate 'words' (signals) of high-density coding capacity. 27975142 6 53 dep read 1093:1096 arg1 ' 1098:1098 ' 1098:1098 ' 1098:1098 Glycan-based 'words' can thus be 'read,' and their message translated into cellular effects by receptors called lectins. 27975142 2 54 theme oligomers 348:356 arg1 variability 327:337 compositional and structural variability 298:337 compositional and structural variability of their oligomers (glycans) 298:366 The extent of compositional and structural variability of their oligomers (glycans) is unsurpassed in Nature due to the unique property of independently combining the following parameters with sequence: anomeric status, linkage positions, ring size, addition of branches and site-specific introduction of substitutions. 27975142 7 55 theme central 1216:1222 arg1 aspects 1224:1230 central aspects 1216:1230 central aspects of the concept of the sugar code 1216:1263 This journal's special issue covers central aspects of the concept of the sugar code. 27975142 7 56 theme sugar 1254:1258 arg1 concept 1239:1245 the concept 1235:1245 the concept of the sugar code 1235:1263 This journal's special issue covers central aspects of the concept of the sugar code. 27057853 10 0 from adiponectin 1520:1530 arg1 blood 1540:1544 cord blood 1535:1544 cord blood 1535:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 7 1 dep detected 1093:1100 arg1 value 1107:1111 m/z value 1362.481-3865.407 1103:1129 m/z value 1362.481-3865.407 1103:1129 A total of 35 N-glycans were detected (m/z value 1362.481-3865.407). 27057853 11 2 theme exploratory 1564:1574 arg1 analysis 1576:1583 a prospective exploratory analysis 1550:1583 a prospective exploratory analysis 1550:1583 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 12 3 theme spectrometry-based 1917:1934 arg1 analysis 1961:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 10 4 theme leptin 1509:1514 arg1 levels 1499:1504 levels 1499:1504 levels of leptin and adiponectin in cord blood 1499:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 3 5 theme birth 541:545 arg1 infants 554:560 120 normal birth weight infants 530:560 120 normal birth weight infants 530:560 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 12 6 theme blood 1941:1945 arg1 analysis 1961:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 2 7 theme environment 268:278 arg1 associations 246:257 associations 246:257 associations of fetal environment and cord blood glycoforms 246:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 1 8 theme predictive 172:181 arg1 factor 183:188 a major predictive factor 164:188 a major predictive factor of type 2 diabetes and cardiovascular disease 164:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 1 8 theme predictive 172:181 arg1 environment 137:147 Fetal environment 131:147 Fetal environment 131:147 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 12 9 theme fucosylation.In 1884:1898 arg1 conclusion 1900:1909 fucosylation.In conclusion 1884:1909 fucosylation.In conclusion 1884:1909 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 4 10 theme matrix-assisted 675:689 arg1 spectrometry 739:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 10 11 theme cord 1535:1538 arg1 blood 1540:1544 cord blood 1535:1544 cord blood 1535:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 7 12 theme m/z 1103:1105 arg1 value 1107:1111 m/z value 1362.481-3865.407 1103:1129 m/z value 1362.481-3865.407 1103:1129 A total of 35 N-glycans were detected (m/z value 1362.481-3865.407). 27057853 9 13 theme m/z 1371:1373 arg1 G3719 1364:1368 G3719 1364:1368 G3719 (m/z value 3719.349) 1364:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 9 13 theme m/z 1371:1373 arg1 value 1375:1379 m/z value 3719.349 1371:1388 m/z value 3719.349 1371:1388 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 4 14 theme comprehensive 626:638 arg1 analysis 660:667 a comprehensive cord blood N-glycan analysis 624:667 a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 624:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 10 15 theme adiponectin 1520:1530 arg1 levels 1499:1504 levels 1499:1504 levels of leptin and adiponectin in cord blood 1499:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 2 16 theme glycoforms 295:304 arg1 associations 246:257 associations 246:257 associations of fetal environment and cord blood glycoforms 246:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 4 17 theme blood 645:649 arg1 analysis 660:667 a comprehensive cord blood N-glycan analysis 624:667 a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 624:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 7 18 theme N-glycans 1078:1086 arg1 total 1066:1070 A total 1064:1070 A total of 35 N-glycans 1064:1086 A total of 35 N-glycans were detected (m/z value 1362.481-3865.407). 27057853 3 19 theme birth 597:601 arg1 cohort 603:608 a longitudinal birth cohort 582:608 a longitudinal birth cohort 582:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 12 20 theme potential 2006:2014 arg1 predictors 2016:2025 potential predictors 2006:2025 potential predictors of a poor fetal environment 2006:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 20 theme potential 2006:2014 arg1 types 1983:1987 5 types 1981:1987 5 types of N-glycans 1981:2000 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 21 theme poor 2032:2035 arg1 environment 2043:2053 a poor fetal environment 2030:2053 a poor fetal environment 2030:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 11 22 theme weight 1709:1714 arg1 gain 1716:1719 6 or 18-month rapid weight gain 1689:1719 6 or 18-month rapid weight gain 1689:1719 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 1 23 theme Fetal 131:135 arg1 environment 137:147 Fetal environment 131:147 Fetal environment 131:147 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 1 23 theme Fetal 131:135 arg1 factor 183:188 a major predictive factor 164:188 a major predictive factor of type 2 diabetes and cardiovascular disease 164:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 3 24 theme normal 534:539 arg1 infants 554:560 120 normal birth weight infants 530:560 120 normal birth weight infants 530:560 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 4 25 theme time-of-flight 719:732 arg1 spectrometry 739:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 9 26 theme G1915 1276:1280 arg1 levels 1266:1271 Abundance levels 1256:1271 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) 1256:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 6 27 theme >0.67 1048:1052 arg1 gain 1042:1045 6 or 18-month rapid weight gain 1015:1045 6 or 18-month rapid weight gain (>0.67 SD score) 1015:1062 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 6 27 theme >0.67 1048:1052 arg1 score 1057:1061 >0.67 SD score 1048:1061 >0.67 SD score 1048:1061 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 12 28 theme N-glycans 1992:2000 arg1 predictors 2016:2025 potential predictors 2006:2025 potential predictors of a poor fetal environment 2006:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 28 theme N-glycans 1992:2000 arg1 types 1983:1987 5 types 1981:1987 5 types of N-glycans 1981:2000 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 29 gly fucosylated 1796:1806 arg1 bi 1808:1809 fucosylated bi 1796:1809 fucosylated bi 1796:1809 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 29 gly fucosylated 1796:1806 arg1 categories 1784:1793 2 different categories 1772:1793 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1772:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 9 30 theme m/z 1283:1285 arg1 G1915 1276:1280 G1915 1276:1280 G1915 (m/z value 1914.698) 1276:1301 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 9 30 theme m/z 1283:1285 arg1 value 1287:1291 m/z value 1914.698 1283:1300 m/z value 1914.698 1283:1300 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 12 31 theme tri-antennary 1814:1826 arg1 N-glycans 1828:1836 tri-antennary N-glycans 1814:1836 tri-antennary N-glycans 1814:1836 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 31 theme tri-antennary 1814:1826 arg1 categories 1784:1793 2 different categories 1772:1793 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1772:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 5 32 theme perinatal 784:792 arg1 outcomes 794:801 perinatal outcomes 784:801 perinatal outcomes 784:801 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 32 theme perinatal 784:792 arg1 levels 850:855 levels 850:855 levels of cord blood leptin and adiponectin 850:892 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 32 theme perinatal 784:792 arg1 small 819:823 small 819:823 small 819:823 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 32 theme perinatal 784:792 arg1 LBW 814:816 LBW 814:816 LBW 814:816 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 2 33 theme cord 284:287 arg1 glycoforms 295:304 cord blood glycoforms 284:304 cord blood glycoforms 284:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 6 34 theme rapid 1029:1033 arg1 gain 1042:1045 6 or 18-month rapid weight gain 1015:1045 6 or 18-month rapid weight gain (>0.67 SD score) 1015:1062 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 6 34 theme rapid 1029:1033 arg1 score 1057:1061 >0.67 SD score 1048:1061 >0.67 SD score 1048:1061 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 3 35 theme perinatal 429:437 arg1 outcomes 439:446 perinatal outcomes 429:446 perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 429:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 0 36 theme Glycosylation 107:119 arg1 Analysis 121:128 A Comprehensive Mass Spectrometry-based Glycosylation Analysis 67:128 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 0:129 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 3 37 from status 380:385 arg1 blood 404:408 neonatal cord blood 390:408 neonatal cord blood 390:408 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 38 dep weight 505:510 arg1 recruited 567:575 recruited 567:575 were recruited from a longitudinal birth cohort 562:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 0 39 from Status 36:41 arg1 Blood 60:64 Neonatal Cord Blood 46:64 Neonatal Cord Blood 46:64 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 0 40 theme Fetal 0:4 arg1 Environment 6:16 Fetal Environment 0:16 Fetal Environment 0:16 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 11 41 dep N-glycans 1604:1612 arg1 G2744 1622:1626 G2744 1622:1626 G2744 1622:1626 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 41 dep N-glycans 1604:1612 arg1 N-glycans 1604:1612 the 5 LBW-related N-glycans 1586:1612 the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) 1586:1652 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 41 dep N-glycans 1604:1612 arg1 G3414 1636:1640 G3414 1636:1640 G3414 1636:1640 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 41 dep N-glycans 1604:1612 arg1 G3719 1647:1651 G3719 1647:1651 G3719 1647:1651 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 41 dep N-glycans 1604:1612 arg1 G3049 1629:1633 G3049 1629:1633 G3049 1629:1633 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 10 42 theme total 1426:1430 arg1 levels 1451:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 3 43 theme poor 461:464 arg1 LBW 513:515 LBW 513:515 LBW 513:515 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 43 theme poor 461:464 arg1 weight 505:510 a poor fetal environment.Thirty-six low birth weight 459:510 a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 459:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 5 44 theme cord 860:863 arg1 blood 865:869 cord blood leptin and adiponectin 860:892 cord blood leptin and adiponectin 860:892 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 45 theme gestational 829:839 arg1 age 841:843 gestational age 829:843 gestational age 829:843 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 12 46 theme different 1774:1782 arg1 bi 1808:1809 fucosylated bi 1796:1809 fucosylated bi 1796:1809 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 46 theme different 1774:1782 arg1 N-glycans 1828:1836 tri-antennary N-glycans 1814:1836 tri-antennary N-glycans 1814:1836 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 46 theme different 1774:1782 arg1 categories 1784:1793 2 different categories 1772:1793 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1772:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 5 47 with Associations 753:764 arg1 outcomes 794:801 perinatal outcomes 784:801 perinatal outcomes 784:801 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 47 with Associations 753:764 arg1 levels 850:855 levels 850:855 levels of cord blood leptin and adiponectin 850:892 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 47 with Associations 753:764 arg1 small 819:823 small 819:823 small 819:823 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 47 with Associations 753:764 arg1 LBW 814:816 LBW 814:816 LBW 814:816 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 3 48 theme neonatal 390:397 arg1 blood 404:408 neonatal cord blood 390:408 neonatal cord blood 390:408 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 49 theme birth 499:503 arg1 LBW 513:515 LBW 513:515 LBW 513:515 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 49 theme birth 499:503 arg1 weight 505:510 a poor fetal environment.Thirty-six low birth weight 459:510 a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 459:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 0 50 theme Neonatal 46:53 arg1 Blood 60:64 Neonatal Cord Blood 46:64 Neonatal Cord Blood 46:64 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 9 51 theme G3049 1332:1336 arg1 levels 1266:1271 Abundance levels 1256:1271 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) 1256:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 10 52 theme abundance 1441:1449 arg1 levels 1451:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 3 53 theme environment.Thirty-six 472:493 arg1 LBW 513:515 LBW 513:515 LBW 513:515 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 53 theme environment.Thirty-six 472:493 arg1 weight 505:510 a poor fetal environment.Thirty-six low birth weight 459:510 a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 459:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 9 54 theme m/z 1339:1341 arg1 G3049 1332:1336 G3049 1332:1336 G3049 (m/z value 3049.105) 1332:1357 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 9 54 theme m/z 1339:1341 arg1 value 1343:1347 m/z value 3049.105 1339:1356 m/z value 3049.105 1339:1356 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 4 55 theme laser-desorption/ionization 691:717 arg1 spectrometry 739:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 5 56 dep blood 865:869 arg1 adiponectin 882:892 adiponectin 882:892 adiponectin 882:892 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 56 dep blood 865:869 arg1 leptin 871:876 leptin 871:876 leptin 871:876 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 11 57 theme prospective 1552:1562 arg1 analysis 1576:1583 a prospective exploratory analysis 1550:1583 a prospective exploratory analysis 1550:1583 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 12 58 theme glycosylation 1947:1959 arg1 analysis 1961:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 2 59 gly glycoforms 295:304 arg1 cord 284:287 cord blood glycoforms 284:304 cord blood glycoforms 284:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 2 59 gly glycoforms 295:304 arg1 blood 289:293 cord blood glycoforms 284:304 cord blood glycoforms 284:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 10 60 from blood 1540:1544 arg1 levels 1499:1504 levels 1499:1504 levels of leptin and adiponectin in cord blood 1499:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 12 61 dep categories 1784:1793 arg1 bi 1808:1809 fucosylated bi 1796:1809 fucosylated bi 1796:1809 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 61 dep categories 1784:1793 arg1 N-glycans 1828:1836 tri-antennary N-glycans 1814:1836 tri-antennary N-glycans 1814:1836 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 61 dep categories 1784:1793 arg1 shows 1970:1974 shows 1970:1974 shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1970:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 61 dep categories 1784:1793 arg1 categories 1784:1793 2 different categories 1772:1793 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1772:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 2 62 theme blood 289:293 arg1 glycoforms 295:304 cord blood glycoforms 284:304 cord blood glycoforms 284:304 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 9 63 theme G3719 1364:1368 arg1 levels 1266:1271 Abundance levels 1256:1271 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) 1256:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 8 64 theme G3414 1163:1167 arg1 levels 1153:1158 abundance levels 1143:1158 abundance levels of G3414 (m/z value 3414.238) 1143:1188 Of these, abundance levels of G3414 (m/z value 3414.238) were inversely correlated with LBW and small for gestational age. 27057853 1 65 theme major 166:170 arg1 factor 183:188 a major predictive factor 164:188 a major predictive factor of type 2 diabetes and cardiovascular disease 164:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 1 65 theme major 166:170 arg1 environment 137:147 Fetal environment 131:147 Fetal environment 131:147 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 8 66 theme m/z 1170:1172 arg1 G3414 1163:1167 G3414 1163:1167 G3414 (m/z value 3414.238) 1163:1188 Of these, abundance levels of G3414 (m/z value 3414.238) were inversely correlated with LBW and small for gestational age. 27057853 8 66 theme m/z 1170:1172 arg1 value 1174:1178 m/z value 3414.238 1170:1187 m/z value 3414.238 1170:1187 Of these, abundance levels of G3414 (m/z value 3414.238) were inversely correlated with LBW and small for gestational age. 27057853 4 67 theme N-glycan 651:658 arg1 analysis 660:667 a comprehensive cord blood N-glycan analysis 624:667 a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 624:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 11 68 theme LBW-related 1592:1602 arg1 G3414 1636:1640 G3414 1636:1640 G3414 1636:1640 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 68 theme LBW-related 1592:1602 arg1 all 1659:1661 all 1659:1661 all 1659:1661 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 68 theme LBW-related 1592:1602 arg1 G2744 1622:1626 G2744 1622:1626 G2744 1622:1626 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 68 theme LBW-related 1592:1602 arg1 G3719 1647:1651 G3719 1647:1651 G3719 1647:1651 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 68 theme LBW-related 1592:1602 arg1 N-glycans 1604:1612 the 5 LBW-related N-glycans 1586:1612 the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) 1586:1652 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 11 68 theme LBW-related 1592:1602 arg1 G3049 1629:1633 G3049 1629:1633 G3049 1629:1633 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 8 69 theme gestational 1239:1249 arg1 age 1251:1253 gestational age 1239:1253 gestational age 1239:1253 Of these, abundance levels of G3414 (m/z value 3414.238) were inversely correlated with LBW and small for gestational age. 27057853 11 70 theme rapid 1703:1707 arg1 gain 1716:1719 6 or 18-month rapid weight gain 1689:1719 6 or 18-month rapid weight gain 1689:1719 In a prospective exploratory analysis, the 5 LBW-related N-glycans (G1915, G2744, G3049, G3414, and G3719) were all inversely associated with 6 or 18-month rapid weight gain. 27057853 3 71 theme weight 547:552 arg1 infants 554:560 120 normal birth weight infants 530:560 120 normal birth weight infants 530:560 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 5 72 theme blood 865:869 arg1 LBW 814:816 LBW 814:816 LBW 814:816 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 5 72 theme blood 865:869 arg1 levels 850:855 levels 850:855 levels of cord blood leptin and adiponectin 850:892 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 3 73 theme longitudinal 584:595 arg1 cohort 603:608 a longitudinal birth cohort 582:608 a longitudinal birth cohort 582:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 1 74 theme diabetes 200:207 arg1 factor 183:188 a major predictive factor 164:188 a major predictive factor of type 2 diabetes and cardiovascular disease 164:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 1 74 theme diabetes 200:207 arg1 environment 137:147 Fetal environment 131:147 Fetal environment 131:147 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 5 75 theme N-glycans 769:777 arg1 Associations 753:764 Associations 753:764 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, 753:893 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 4 76 theme cord 640:643 arg1 analysis 660:667 a comprehensive cord blood N-glycan analysis 624:667 a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 624:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 12 77 theme tetra-antennary 1850:1864 arg1 N-glycans 1866:1874 tetra-antennary N-glycans 1850:1874 tetra-antennary N-glycans 1850:1874 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 9 78 theme Abundance 1256:1264 arg1 levels 1266:1271 Abundance levels 1256:1271 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) 1256:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 2 79 theme fetal 262:266 arg1 environment 268:278 fetal environment 262:278 fetal environment 262:278 However, associations of fetal environment and cord blood glycoforms are uncertain. 27057853 1 80 theme type 193:196 arg1 diabetes 200:207 type 2 diabetes 193:207 type 2 diabetes 193:207 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 5 81 theme logistic 916:923 arg1 regression 937:946 logistic or multiple regression 916:946 regression 937:946 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 6 82 theme SD 1054:1055 arg1 gain 1042:1045 6 or 18-month rapid weight gain 1015:1045 6 or 18-month rapid weight gain (>0.67 SD score) 1015:1062 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 6 82 theme SD 1054:1055 arg1 score 1057:1061 >0.67 SD score 1048:1061 >0.67 SD score 1048:1061 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 1 83 theme cardiovascular 213:226 arg1 disease 228:234 cardiovascular disease 213:234 cardiovascular disease 213:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 0 84 theme Comprehensive 69:81 arg1 Analysis 121:128 A Comprehensive Mass Spectrometry-based Glycosylation Analysis 67:128 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 0:129 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 5 85 theme multiple 928:935 arg1 regression 937:946 logistic or multiple regression 916:946 regression 937:946 Associations of N-glycans with perinatal outcomes, including LBW, small for gestational age, and levels of cord blood leptin and adiponectin, were evaluated using logistic or multiple regression. 27057853 0 86 from Environment 6:16 arg1 Blood 60:64 Neonatal Cord Blood 46:64 Neonatal Cord Blood 46:64 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 4 87 theme mass 734:737 arg1 spectrometry 739:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry 675:750 We conducted a comprehensive cord blood N-glycan analysis using matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. 27057853 0 88 theme Spectrometry-based 88:105 arg1 Analysis 121:128 A Comprehensive Mass Spectrometry-based Glycosylation Analysis 67:128 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 0:129 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 6 89 theme weight 1035:1040 arg1 gain 1042:1045 6 or 18-month rapid weight gain 1015:1045 6 or 18-month rapid weight gain (>0.67 SD score) 1015:1062 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 6 89 theme weight 1035:1040 arg1 score 1057:1061 >0.67 SD score 1048:1061 >0.67 SD score 1048:1061 We also prospectively explored correlations between N-glycans and 6 or 18-month rapid weight gain (>0.67 SD score). 27057853 12 90 theme fucosylated 1796:1806 arg1 bi 1808:1809 fucosylated bi 1796:1809 fucosylated bi 1796:1809 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 90 theme fucosylated 1796:1806 arg1 categories 1784:1793 2 different categories 1772:1793 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment 1772:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 9 91 theme G2744 1304:1308 arg1 levels 1266:1271 Abundance levels 1256:1271 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) 1256:1389 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 0 92 theme Glycosylation 22:34 arg1 Status 36:41 Glycosylation Status 22:41 Glycosylation Status 22:41 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 9 93 theme m/z 1311:1313 arg1 value 1315:1319 m/z value 2743.994 1311:1328 m/z value 2743.994 1311:1328 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 9 93 theme m/z 1311:1313 arg1 G2744 1304:1308 G2744 1304:1308 G2744 (m/z value 2743.994) 1304:1329 Abundance levels of G1915 (m/z value 1914.698), G2744 (m/z value 2743.994), G3049 (m/z value 3049.105), and G3719 (m/z value 3719.349) were inversely related to LBW. 27057853 3 94 theme glycosylation 366:378 arg1 status 380:385 glycosylation status 366:385 glycosylation status in neonatal cord blood 366:408 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 95 theme fetal 466:470 arg1 LBW 513:515 LBW 513:515 LBW 513:515 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 95 theme fetal 466:470 arg1 weight 505:510 a poor fetal environment.Thirty-six low birth weight 459:510 a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 459:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 12 96 theme environment 2043:2053 arg1 predictors 2016:2025 potential predictors 2006:2025 potential predictors of a poor fetal environment 2006:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 12 96 theme environment 2043:2053 arg1 types 1983:1987 5 types 1981:1987 5 types of N-glycans 1981:2000 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 0 97 theme Cord 55:58 arg1 Blood 60:64 Neonatal Cord Blood 46:64 Neonatal Cord Blood 46:64 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 12 98 theme fetal 2037:2041 arg1 environment 2043:2053 a poor fetal environment 2030:2053 a poor fetal environment 2030:2053 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27057853 3 99 theme cord 399:402 arg1 blood 404:408 neonatal cord blood 390:408 neonatal cord blood 390:408 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 10 100 from levels 1499:1504 arg1 blood 1540:1544 cord blood 1535:1544 cord blood 1535:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 10 101 theme N-glycan 1432:1439 arg1 levels 1451:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels 1422:1456 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 3 102 theme low 495:497 arg1 LBW 513:515 LBW 513:515 LBW 513:515 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 3 102 theme low 495:497 arg1 weight 505:510 a poor fetal environment.Thirty-six low birth weight 459:510 a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort 459:608 In this study, we aimed to determine whether glycosylation status in neonatal cord blood is associated with perinatal outcomes reflecting a poor fetal environment.Thirty-six low birth weight (LBW) infants and 120 normal birth weight infants were recruited from a longitudinal birth cohort. 27057853 1 103 theme disease 228:234 arg1 factor 183:188 a major predictive factor 164:188 a major predictive factor of type 2 diabetes and cardiovascular disease 164:234 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 1 103 theme disease 228:234 arg1 environment 137:147 Fetal environment 131:147 Fetal environment 131:147 Fetal environment is known to be a major predictive factor of type 2 diabetes and cardiovascular disease. 27057853 8 104 theme abundance 1143:1151 arg1 levels 1153:1158 abundance levels 1143:1158 abundance levels of G3414 (m/z value 3414.238) 1143:1188 Of these, abundance levels of G3414 (m/z value 3414.238) were inversely correlated with LBW and small for gestational age. 27057853 10 105 from leptin 1509:1514 arg1 blood 1540:1544 cord blood 1535:1544 cord blood 1535:1544 The total N-glycan abundance levels were strongly positively correlated with levels of leptin and adiponectin in cord blood. 27057853 0 106 dep Environment 6:16 arg1 Analysis 121:128 A Comprehensive Mass Spectrometry-based Glycosylation Analysis 67:128 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 0:129 Fetal Environment and Glycosylation Status in Neonatal Cord Blood: A Comprehensive Mass Spectrometry-based Glycosylation Analysis. 27057853 12 107 theme cord 1936:1939 arg1 analysis 1961:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 mass spectrometry-based cord blood glycosylation analysis 1912:1968 These N-glycans are structurally categorized into 2 different categories: fucosylated bi or tri-antennary N-glycans; and tri or tetra-antennary N-glycans without fucosylation.In conclusion, mass spectrometry-based cord blood glycosylation analysis shows that 5 types of N-glycans are potential predictors of a poor fetal environment. 27984785 6 0 theme Ebola 924:928 arg1 virus 930:934 Ebola virus 924:934 Ebola virus 924:934 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 7 1 theme entry 1124:1128 arg1 studies 1149:1155 future ebolaviral entry and immunogenicity studies 1106:1155 future ebolaviral entry and immunogenicity studies 1106:1155 The data presented here should serve as the foundation for future ebolaviral entry and immunogenicity studies. 27984785 1 2 theme hemorrhagic 158:168 arg1 fevers 170:175 severe and often fatal viral hemorrhagic fevers 129:175 severe and often fatal viral hemorrhagic fevers 129:175 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 4 3 theme tri- 582:585 arg1 glycans 616:622 bi-, tri-, and tetra-antennary complex glycans 577:622 bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 577:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 6 4 theme virus 930:934 arg1 variant 981:987 the Makona variant 970:987 the Makona variant 970:987 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 4 theme virus 930:934 arg1 variant 958:964 the original Yambuku variant 937:964 the original Yambuku variant 937:964 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 4 theme virus 930:934 arg1 variants 912:919 two variants 908:919 two variants of Ebola virus 908:934 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 5 dep amount 767:772 arg1 the 763:765 the 763:765 the 763:765 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 5 6 theme ebolavirus 731:740 arg1 GP1,2s 742:747 the different ebolavirus GP1,2s 717:747 the different ebolavirus GP1,2s 717:747 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 6 7 theme ebolavirus 828:837 arg1 GP1,2s. 839:845 ebolavirus GP1,2s. 828:845 ebolavirus GP1,2s. 828:845 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 7 8 theme future 1106:1111 arg1 studies 1149:1155 future ebolaviral entry and immunogenicity studies 1106:1155 future ebolaviral entry and immunogenicity studies 1106:1155 The data presented here should serve as the foundation for future ebolaviral entry and immunogenicity studies. 27984785 6 9 theme responsible 989:999 arg1 variants 912:919 two variants 908:919 two variants of Ebola virus 908:934 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 3 10 theme ebolaviruses 367:378 arg1 unknown 392:398 unknown 392:398 unknown 392:398 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 10 theme ebolaviruses 367:378 arg1 patterns 355:362 the precise glycosylation patterns 329:362 the precise glycosylation patterns of ebolaviruses 329:378 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 7 11 theme ebolaviral 1113:1122 arg1 studies 1149:1155 future ebolaviral entry and immunogenicity studies 1106:1155 future ebolaviral entry and immunogenicity studies 1106:1155 The data presented here should serve as the foundation for future ebolaviral entry and immunogenicity studies. 27984785 0 12 gly glycoproteins 67:79 arg1 glycoproteins 67:79 ebolavirus glycoproteins 56:79 ebolavirus glycoproteins 56:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 3 13 theme glycosylation 341:353 arg1 unknown 392:398 unknown 392:398 unknown 392:398 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 13 theme glycosylation 341:353 arg1 patterns 355:362 the precise glycosylation patterns 329:362 the precise glycosylation patterns of ebolaviruses 329:378 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 6 14 theme structures 795:804 arg1 type 778:781 type 778:781 type 778:781 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 14 theme structures 795:804 arg1 structures 795:804 O-glycan structures 786:804 O-glycan structures 786:804 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 14 theme structures 795:804 arg1 amount 767:772 amount 767:772 amount 767:772 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 4 15 located present 477:483 arg1 GP1,2 488:492 GP1,2 488:492 GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 488:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 15 located present 477:483 arg2 structures 462:471 approximately 50 different N-glycan structures 426:471 approximately 50 different N-glycan structures 426:471 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 3 16 theme ebolavirus 259:268 arg1 GP1,2 293:297 GP1,2 293:297 GP1,2 293:297 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 16 theme ebolavirus 259:268 arg1 glycoprotein 279:290 The ebolavirus envelope glycoprotein 255:290 The ebolavirus envelope glycoprotein (GP1,2) 255:298 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 6 17 theme O-glycan 786:793 arg1 structures 795:804 O-glycan structures 786:804 O-glycan structures 786:804 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 3 18 gly glycoprotein 279:290 arg1 GP1,2 293:297 GP1,2 293:297 GP1,2 293:297 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 18 gly glycoprotein 279:290 arg1 glycoprotein 279:290 The ebolavirus envelope glycoprotein 255:290 The ebolavirus envelope glycoprotein (GP1,2) 255:298 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 0 19 theme glycoproteins 67:79 arg1 patterns 44:51 N- and O-linked glycosylation patterns 14:51 N- and O-linked glycosylation patterns of ebolavirus glycoproteins 14:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 2 20 from disease 236:242 arg1 humans 247:252 humans 247:252 humans 247:252 Four distinct ebolaviruses are known to cause Ebola virus disease in humans. 27984785 4 21 attach present 477:483 arg1 GP1,2 488:492 GP1,2 488:492 GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 488:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 21 attach present 477:483 arg2 structures 462:471 approximately 50 different N-glycan structures 426:471 approximately 50 different N-glycan structures 426:471 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 6 22 theme Makona 974:979 arg1 variant 981:987 the Makona variant 970:987 the Makona variant 970:987 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 4 23 from GP1,2 488:492 arg1 present 477:483 present 477:483 present 477:483 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 1 24 theme emerging 99:106 arg1 Ebolaviruses 82:93 Ebolaviruses 82:93 Ebolaviruses 82:93 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 1 24 theme emerging 99:106 arg1 pathogens 108:116 emerging pathogens 99:116 emerging pathogens that cause severe and often fatal viral hemorrhagic fevers 99:175 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 0 25 link O-linked 21:28 arg1 patterns 44:51 N- and O-linked glycosylation patterns 14:51 N- and O-linked glycosylation patterns of ebolavirus glycoproteins 14:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 0 26 theme N- 14:15 arg1 patterns 44:51 N- and O-linked glycosylation patterns 14:51 N- and O-linked glycosylation patterns of ebolavirus glycoproteins 14:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 4 27 theme bi- 577:579 arg1 glycans 616:622 bi-, tri-, and tetra-antennary complex glycans 577:622 bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 577:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 28 from present 477:483 arg1 GP1,2 488:492 GP1,2 488:492 GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 488:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 0 29 theme glycosylation 30:42 arg1 patterns 44:51 N- and O-linked glycosylation patterns 14:51 N- and O-linked glycosylation patterns of ebolavirus glycoproteins 14:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 3 30 theme envelope 270:277 arg1 GP1,2 293:297 GP1,2 293:297 GP1,2 293:297 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 30 theme envelope 270:277 arg1 glycoprotein 279:290 The ebolavirus envelope glycoprotein 255:290 The ebolavirus envelope glycoprotein (GP1,2) 255:298 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 2 31 theme distinct 183:190 arg1 ebolaviruses 192:203 Four distinct ebolaviruses 178:203 Four distinct ebolaviruses 178:203 Four distinct ebolaviruses are known to cause Ebola virus disease in humans. 27984785 5 32 theme different 721:729 arg1 GP1,2s 742:747 the different ebolavirus GP1,2s 717:747 the different ebolavirus GP1,2s 717:747 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 0 33 theme O-linked 21:28 arg1 patterns 44:51 N- and O-linked glycosylation patterns 14:51 N- and O-linked glycosylation patterns of ebolavirus glycoproteins 14:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 4 34 theme high 551:554 arg1 mannose 556:562 high mannose 551:562 high mannose 551:562 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 3 35 gly glycosylation 341:353 arg1 ebolaviruses 367:378 ebolaviruses 367:378 ebolaviruses 367:378 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 6 36 theme O-glycan 861:868 arg1 dissimilarity 870:882 this O-glycan dissimilarity 856:882 this O-glycan dissimilarity 856:882 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 6 37 theme epidemic 1037:1044 arg1 Western 1021:1027 the most recent Western 1005:1027 the most recent Western African epidemic 1005:1044 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 7 38 theme immunogenicity 1134:1147 arg1 studies 1149:1155 future ebolaviral entry and immunogenicity studies 1106:1155 future ebolaviral entry and immunogenicity studies 1106:1155 The data presented here should serve as the foundation for future ebolaviral entry and immunogenicity studies. 27984785 6 39 theme Yambuku 950:956 arg1 variant 958:964 the original Yambuku variant 937:964 the original Yambuku variant 937:964 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 5 40 theme overall 669:675 arg1 composition 686:696 The overall N-glycan composition 665:696 The overall N-glycan composition 665:696 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 5 40 theme overall 669:675 arg1 similar 701:707 similar 701:707 similar 701:707 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 3 41 theme precise 333:339 arg1 unknown 392:398 unknown 392:398 unknown 392:398 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 41 theme precise 333:339 arg1 patterns 355:362 the precise glycosylation patterns 329:362 the precise glycosylation patterns of ebolaviruses 329:378 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 2 42 theme virus 230:234 arg1 disease 236:242 Ebola virus disease 224:242 Ebola virus disease in humans 224:252 Four distinct ebolaviruses are known to cause Ebola virus disease in humans. 27984785 0 43 theme patterns 44:51 arg1 Comparison 0:9 Comparison 0:9 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 0:80 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 5 44 theme N-glycan 677:684 arg1 composition 686:696 The overall N-glycan composition 665:696 The overall N-glycan composition 665:696 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 5 44 theme N-glycan 677:684 arg1 similar 701:707 similar 701:707 similar 701:707 The overall N-glycan composition is similar between the different ebolavirus GP1,2s. 27984785 2 45 theme Ebola 224:228 arg1 disease 236:242 Ebola virus disease 224:242 Ebola virus disease in humans 224:252 Four distinct ebolaviruses are known to cause Ebola virus disease in humans. 27984785 1 46 theme severe 129:134 arg1 fevers 170:175 severe and often fatal viral hemorrhagic fevers 129:175 severe and often fatal viral hemorrhagic fevers 129:175 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 4 47 theme complex 608:614 arg1 glycans 616:622 bi-, tri-, and tetra-antennary complex glycans 577:622 bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 577:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 48 theme N-glycan 453:460 arg1 structures 462:471 approximately 50 different N-glycan structures 426:471 approximately 50 different N-glycan structures 426:471 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 0 49 gly glycosylation 30:42 arg1 glycoproteins 67:79 ebolavirus glycoproteins 56:79 ebolavirus glycoproteins 56:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 4 50 attach derived 494:500 arg1 mannose 556:562 high mannose 551:562 high mannose 551:562 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 50 attach derived 494:500 arg2 GP1,2 488:492 GP1,2 488:492 GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 488:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 50 attach derived 494:500 arg1 ebolaviruses 527:538 the four pathogenic ebolaviruses 507:538 the four pathogenic ebolaviruses 507:538 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 50 attach derived 494:500 arg1 hybrid 565:570 hybrid 565:570 hybrid 565:570 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 50 attach derived 494:500 arg1 glycans 616:622 bi-, tri-, and tetra-antennary complex glycans 577:622 bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 577:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 51 theme tetra-antennary 592:606 arg1 complex 608:614 tetra-antennary complex 592:614 tetra-antennary complex 592:614 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 3 52 gly glycosylated 311:322 arg1 GP1,2 293:297 GP1,2 293:297 GP1,2 293:297 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 3 52 gly glycosylated 311:322 arg1 glycoprotein 279:290 The ebolavirus envelope glycoprotein 255:290 The ebolavirus envelope glycoprotein (GP1,2) 255:298 The ebolavirus envelope glycoprotein (GP1,2) is heavily glycosylated, but the precise glycosylation patterns of ebolaviruses are largely unknown. 27984785 4 53 theme different 443:451 arg1 structures 462:471 approximately 50 different N-glycan structures 426:471 approximately 50 different N-glycan structures 426:471 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 1 54 theme fatal 146:150 arg1 fevers 170:175 severe and often fatal viral hemorrhagic fevers 129:175 severe and often fatal viral hemorrhagic fevers 129:175 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 4 55 theme sialic 652:657 arg1 acid 659:662 sialic acid 652:662 sialic acid 652:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 6 56 theme recent 1014:1019 arg1 Western 1021:1027 the most recent Western 1005:1027 the most recent Western African epidemic 1005:1044 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 27984785 0 57 theme ebolavirus 56:65 arg1 glycoproteins 67:79 ebolavirus glycoproteins 56:79 ebolavirus glycoproteins 56:79 Comparison of N- and O-linked glycosylation patterns of ebolavirus glycoproteins. 27984785 4 58 theme pathogenic 516:525 arg1 mannose 556:562 high mannose 551:562 high mannose 551:562 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 58 theme pathogenic 516:525 arg1 glycans 616:622 bi-, tri-, and tetra-antennary complex glycans 577:622 bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid 577:662 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 58 theme pathogenic 516:525 arg1 ebolaviruses 527:538 the four pathogenic ebolaviruses 507:538 the four pathogenic ebolaviruses 507:538 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 4 58 theme pathogenic 516:525 arg1 hybrid 565:570 hybrid 565:570 hybrid 565:570 Here we demonstrate that approximately 50 different N-glycan structures are present in GP1,2 derived from the four pathogenic ebolaviruses, including high mannose, hybrid, and bi-, tri-, and tetra-antennary complex glycans with and without fucose and sialic acid. 27984785 1 59 theme viral 152:156 arg1 fevers 170:175 severe and often fatal viral hemorrhagic fevers 129:175 severe and often fatal viral hemorrhagic fevers 129:175 Ebolaviruses are emerging pathogens that cause severe and often fatal viral hemorrhagic fevers. 27984785 6 60 theme original 941:948 arg1 variant 958:964 the original Yambuku variant 937:964 the original Yambuku variant 937:964 In contrast, the amount and type of O-glycan structures varies widely between ebolavirus GP1,2s. Notably, this O-glycan dissimilarity is also present between two variants of Ebola virus, the original Yambuku variant and the Makona variant responsible for the most recent Western African epidemic. 28518173 7 0 theme mass 1377:1380 arg1 MS 1396:1397 MS 1396:1397 MS 1396:1397 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 0 theme mass 1377:1380 arg1 spectrometry 1382:1393 (v) mass spectrometry 1373:1393 (v) mass spectrometry (MS) analysis 1373:1407 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 6 1 theme O-glycans 845:853 arg1 analysis 819:826 quantitative analysis 806:826 quantitative analysis of N-glycans and O-glycans from a single specimen 806:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 4 2 theme enzymatic 548:556 arg1 reactions 571:579 these enzymatic and chemical reactions 542:579 these enzymatic and chemical reactions 542:579 However, it is challenging to combine these enzymatic and chemical reactions in order to analyze both N- and O-glycans. 28518173 6 3 from analysis 819:826 arg1 specimen 869:876 a single specimen 860:876 a single specimen 860:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 7 4 theme aldehyde-active 1044:1058 arg1 support 1066:1072 an aldehyde-active solid support 1041:1072 an aldehyde-active solid support 1041:1072 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 2 5 theme asparagine 258:267 arg1 chains 274:279 asparagine side chains 258:279 asparagine side chains 258:279 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 6 6 theme N-glycans 831:839 arg1 analysis 819:826 quantitative analysis 806:826 quantitative analysis of N-glycans and O-glycans from a single specimen 806:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 2 7 dep atom 239:242 arg1 N-linked 245:252 N-linked 245:252 N-linked 245:252 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 7 8 dep includes 987:994 arg1 release 1222:1228 (iv) release 1217:1228 (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans 1217:1370 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 8 dep includes 987:994 arg1 protocol 973:980 a step-by-step GIG protocol 954:980 a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) 954:1417 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 8 dep includes 987:994 arg1 analysis 1400:1407 (v) mass spectrometry (MS) analysis 1373:1407 (v) mass spectrometry (MS) analysis 1373:1407 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 5 9 theme N- 757:758 arg1 analysis 745:752 the simultaneous analysis 728:752 the simultaneous analysis of N- and O-glycans on a solid support 728:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 8 10 theme tool 1578:1581 arg1 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 11 from ions 1618:1621 arg1 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 12 theme protein 1015:1021 arg1 immobilization 1023:1036 (i) protein immobilization 1011:1036 (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion 1011:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 13 theme precursor 1592:1600 arg1 masses 1602:1607 precursor masses 1592:1607 precursor masses 1592:1607 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 5 14 theme n 660:660 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 2 15 theme chains 274:279 arg1 atom 239:242 the nitrogen atom 226:242 the nitrogen atom (N-linked) of asparagine side chains 226:279 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 7 16 theme GIG 1498:1500 arg1 software 1488:1495 in-house developed software 1469:1495 in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool) 1469:1567 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 16 theme GIG 1498:1500 arg1 Tool 1502:1505 GIG Tool 1498:1505 GIG Tool; free to download via http 1498:1532 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 5 17 theme for 677:679 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 8 18 theme relative 1866:1873 arg1 abundance 1875:1883 the relative abundance 1862:1883 the relative abundance of N-glycans 1862:1896 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 19 theme glycan 1799:1804 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 20 theme isobaric 1782:1789 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 21 theme glycan 1627:1632 arg1 fragments 1634:1642 glycan fragments 1627:1642 glycan fragments 1627:1642 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 22 theme in-house 1469:1476 arg1 software 1488:1495 in-house developed software 1469:1495 in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool) 1469:1567 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 22 theme in-house 1469:1476 arg1 Tool 1502:1505 GIG Tool 1498:1505 GIG Tool; free to download via http 1498:1532 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 23 theme for 1795:1797 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 24 theme glycans 1455:1461 arg1 identification 1437:1450 identification 1437:1450 identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool) 1437:1567 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 25 theme O-glycans 1233:1241 arg1 release 1222:1228 (iv) release 1217:1228 (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans 1217:1370 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 0 26 theme solid-phase 56:66 arg1 method 68:73 a solid-phase method 54:73 a solid-phase method 54:73 Simultaneous quantification of N- and O-glycans using a solid-phase method. 28518173 2 27 theme nitrogen 230:237 arg1 atom 239:242 the nitrogen atom 226:242 the nitrogen atom (N-linked) of asparagine side chains 226:279 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 8 28 contain containing 1771:1780 arg1 amine 1765:1769 quaternary amine 1754:1769 quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1754:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 28 contain containing 1771:1780 arg2 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 29 theme oxonium 1610:1616 arg1 ions 1618:1621 oxonium ions 1610:1621 oxonium ions 1610:1621 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 30 theme carbodiimide 1144:1155 arg1 coupling 1157:1164 carbodiimide coupling 1144:1164 carbodiimide coupling 1144:1164 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 31 dep Tool 1502:1505 arg1 //www.biomarkercenter.org/gigtool 1534:1566 //www.biomarkercenter.org/gigtool 1534:1566 //www.biomarkercenter.org/gigtool 1534:1566 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 32 theme chromatography 1664:1677 arg1 -MS/MS 1683:1688 liquid chromatography (LC)-MS/MS 1657:1688 liquid chromatography (LC)-MS/MS 1657:1688 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 32 theme chromatography 1664:1677 arg1 spectra 1696:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra 1649:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification 1649:1728 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 33 theme glycan 1708:1713 arg1 identification 1715:1728 glycan identification 1708:1728 glycan identification 1708:1728 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 34 dep masses 1602:1607 arg1 amine 1765:1769 quaternary amine 1754:1769 quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1754:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 5 35 theme extraction 688:697 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 6 36 theme automated 918:926 arg1 platform 928:935 a high-throughput automated platform 900:935 a high-throughput automated platform 900:935 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 8 37 used used 1835:1838 arg2 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 38 dep provide 946:952 arg1 data 1419:1422 data 1419:1422 data 1419:1422 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 5 39 theme GIG 700:702 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 2 40 dep atom 298:301 arg1 O-linked 304:311 O-linked 304:311 O-linked 304:311 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 2 41 theme side 338:341 arg1 chains 343:348 side chains 338:348 side chains 338:348 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 0 42 theme Simultaneous 0:11 arg1 quantification 13:26 Simultaneous quantification 0:26 Simultaneous quantification of N- and O-glycans 0:46 Simultaneous quantification of N- and O-glycans using a solid-phase method. 28518173 8 43 theme mass 1691:1694 arg1 -MS/MS 1683:1688 liquid chromatography (LC)-MS/MS 1657:1688 liquid chromatography (LC)-MS/MS 1657:1688 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 43 theme mass 1691:1694 arg1 spectra 1696:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra 1649:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification 1649:1728 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 2 44 theme protein 374:380 arg1 forms 396:400 the two major protein glycosylation forms 360:400 the two major protein glycosylation forms 360:400 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 5 45 theme O-glycans 764:772 arg1 analysis 745:752 the simultaneous analysis 728:752 the simultaneous analysis of N- and O-glycans on a solid support 728:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 6 46 theme quantitative 806:817 arg1 analysis 819:826 quantitative analysis 806:826 quantitative analysis of N-glycans and O-glycans from a single specimen 806:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 7 47 dep release 1173:1179 arg1 iii 1168:1170 iii 1168:1170 iii 1168:1170 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 48 theme free 1508:1511 arg1 software 1488:1495 in-house developed software 1469:1495 in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool) 1469:1567 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 48 theme free 1508:1511 arg1 Tool 1502:1505 GIG Tool 1498:1505 GIG Tool; free to download via http 1498:1532 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 49 from spectra 1696:1702 arg1 masses 1602:1607 precursor masses 1592:1607 precursor masses 1592:1607 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 49 from spectra 1696:1702 arg1 ions 1618:1621 oxonium ions 1610:1621 oxonium ions 1610:1621 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 49 from spectra 1696:1702 arg1 fragments 1634:1642 glycan fragments 1627:1642 glycan fragments 1627:1642 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 49 from spectra 1696:1702 arg1 ions 1744:1747 reporter ions 1735:1747 reporter ions 1735:1747 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 9 50 theme GIG 1903:1905 arg1 protocol 1907:1914 The GIG protocol 1899:1914 The GIG protocol 1899:1914 The GIG protocol takes ∼3 d. 28518173 1 51 dep structure 169:177 arg1 the 165:167 the 165:167 the 165:167 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 8 52 theme abundance 1875:1883 arg1 quantification 1844:1857 quantification 1844:1857 quantification of the relative abundance of N-glycans 1844:1896 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 53 theme reductive 1077:1085 arg1 amination 1087:1095 reductive amination 1077:1095 reductive amination 1077:1095 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 54 theme PNGase 1197:1202 arg1 digestion 1206:1214 PNGase F digestion 1197:1214 PNGase F digestion 1197:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 55 theme spectrometry 1382:1393 arg1 analysis 1400:1407 (v) mass spectrometry (MS) analysis 1373:1407 (v) mass spectrometry (MS) analysis 1373:1407 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 56 theme N-glycans 1184:1192 arg1 immobilization 1023:1036 (i) protein immobilization 1011:1036 (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion 1011:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 6 57 theme single 862:867 arg1 specimen 869:876 a single specimen 860:876 a single specimen 860:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 7 58 theme solid 1060:1064 arg1 support 1066:1072 an aldehyde-active solid support 1041:1072 an aldehyde-active solid support 1041:1072 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 59 dep spectrometry 1382:1393 arg1 v 1374:1374 v 1374:1374 v 1374:1374 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 2 60 theme side 269:272 arg1 chains 274:279 asparagine side chains 258:279 asparagine side chains 258:279 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 7 61 theme alditol 1341:1347 arg1 peeling 1349:1355 alditol peeling 1341:1355 alditol peeling 1341:1355 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 3 62 theme chemical 485:492 arg1 reaction 494:501 chemical reaction 485:501 chemical reaction 485:501 N-glycans can be released by glycosidases, whereas O-glycans are often cleaved by chemical reaction. 28518173 8 63 theme N-glycans 1888:1896 arg1 abundance 1875:1883 the relative abundance 1862:1883 the relative abundance of N-glycans 1862:1896 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 64 theme GIG 1574:1576 arg1 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 6 65 theme high-throughput 902:916 arg1 platform 928:935 a high-throughput automated platform 900:935 a high-throughput automated platform 900:935 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 8 66 theme reporter 1735:1742 arg1 ions 1744:1747 reporter ions 1735:1747 reporter ions 1735:1747 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 5 67 theme glycoprotei 648:658 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 7 68 dep 1-phenyl-3-methyl-5-pyrazolone 1293:1322 arg1 prevent 1333:1339 prevent 1333:1339 to prevent alditol peeling from O-glycans 1330:1370 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 69 theme 1-phenyl-3-methyl-5-pyrazolone 1293:1322 arg1 presence 1281:1288 the presence 1277:1288 the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans 1277:1370 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 5 70 theme immobilization 662:675 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 5 71 theme solid 779:783 arg1 support 785:791 a solid support 777:791 a solid support 777:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 7 72 theme developed 1478:1486 arg1 software 1488:1495 in-house developed software 1469:1495 in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool) 1469:1567 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 72 theme developed 1478:1486 arg1 Tool 1502:1505 GIG Tool 1498:1505 GIG Tool; free to download via http 1498:1532 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 73 dep immobilization 1023:1036 arg1 release 1173:1179 (iii) release 1167:1179 (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion 1011:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 73 dep immobilization 1023:1036 arg1 stabilization 1103:1115 (ii) stabilization 1098:1115 (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion 1011:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 73 dep immobilization 1023:1036 arg1 i 1012:1012 i 1012:1012 i 1012:1012 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 7 74 theme sialic 1128:1133 arg1 acids 1135:1139 fragile sialic acids 1120:1139 fragile sialic acids 1120:1139 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 75 theme tag 1791:1793 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 76 theme step-by-step 956:967 arg1 protocol 973:980 a step-by-step GIG protocol 954:980 a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) 954:1417 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 77 theme quaternary 1754:1763 arg1 amine 1765:1769 quaternary amine 1754:1769 quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1754:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 1 78 contain has 90:92 arg2 role 104:107 a pivotal role 94:107 a pivotal role 94:107 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 1 78 contain has 90:92 arg1 Glycosylation 76:88 Glycosylation 76:88 Glycosylation 76:88 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 7 79 theme GIG 969:971 arg1 protocol 973:980 a step-by-step GIG protocol 954:980 a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) 954:1417 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 1 80 theme diverse 114:120 arg1 range 122:126 a diverse range 112:126 a diverse range of biological activities 112:151 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 8 81 theme QUANTITY 1807:1814 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 82 theme acids 1135:1139 arg1 stabilization 1103:1115 (ii) stabilization 1098:1115 (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion 1011:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 83 theme isobaric 1817:1824 arg1 tags 1826:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 isobaric tag for glycan (QUANTITY) isobaric tags 1782:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 5 84 from analysis 745:752 arg1 support 785:791 a solid support 777:791 a solid support 777:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 8 85 theme tandem 1649:1654 arg1 -MS/MS 1683:1688 liquid chromatography (LC)-MS/MS 1657:1688 liquid chromatography (LC)-MS/MS 1657:1688 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 85 theme tandem 1649:1654 arg1 spectra 1696:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra 1649:1702 tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification 1649:1728 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 1 86 theme activities 142:151 arg1 range 122:126 a diverse range 112:126 a diverse range of biological activities 112:151 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 5 87 theme glycan 681:686 arg1 method 705:710 a glycoprotei n immobilization for glycan extraction (GIG) method 646:710 a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support 646:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 8 88 theme liquid 1657:1662 arg1 LC 1680:1681 LC 1680:1681 LC 1680:1681 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 8 88 theme liquid 1657:1662 arg1 chromatography 1664:1677 liquid chromatography 1657:1677 liquid chromatography (LC)-MS/MS 1657:1688 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 89 dep stabilization 1103:1115 arg1 ii 1099:1100 ii 1099:1100 ii 1099:1100 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 2 90 dep serine 317:322 arg1 chains 343:348 side chains 338:348 side chains 338:348 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 2 91 theme threonine 328:336 arg1 atom 298:301 the oxygen atom 287:301 the oxygen atom (O-linked) of serine and threonine side chains 287:348 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 0 92 theme N- 31:32 arg1 quantification 13:26 Simultaneous quantification 0:26 Simultaneous quantification of N- and O-glycans 0:46 Simultaneous quantification of N- and O-glycans using a solid-phase method. 28518173 1 93 theme pivotal 96:102 arg1 role 104:107 a pivotal role 94:107 a pivotal role 94:107 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 2 94 theme glycosylation 382:394 arg1 forms 396:400 the two major protein glycosylation forms 360:400 the two major protein glycosylation forms 360:400 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 7 95 from immobilization 1023:1036 arg1 support 1066:1072 an aldehyde-active solid support 1041:1072 an aldehyde-active solid support 1041:1072 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 2 96 theme major 368:372 arg1 forms 396:400 the two major protein glycosylation forms 360:400 the two major protein glycosylation forms 360:400 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 0 97 theme O-glycans 38:46 arg1 quantification 13:26 Simultaneous quantification 0:26 Simultaneous quantification of N- and O-glycans 0:46 Simultaneous quantification of N- and O-glycans using a solid-phase method. 28518173 1 98 theme proteins 195:202 arg1 structure 169:177 structure 169:177 structure 169:177 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 1 98 theme proteins 195:202 arg1 function 183:190 function 183:190 function 183:190 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 7 99 theme F 1204:1204 arg1 digestion 1206:1214 PNGase F digestion 1197:1214 PNGase F digestion 1197:1214 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 8 100 from ions 1744:1747 arg1 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 7 101 theme fragile 1120:1126 arg1 acids 1135:1139 fragile sialic acids 1120:1139 fragile sialic acids 1120:1139 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 5 102 theme simultaneous 732:743 arg1 analysis 745:752 the simultaneous analysis 728:752 the simultaneous analysis of N- and O-glycans on a solid support 728:791 We recently developed a glycoprotei n immobilization for glycan extraction (GIG) method that allows for the simultaneous analysis of N- and O-glycans on a solid support. 28518173 7 103 dep release 1222:1228 arg1 iv 1218:1219 iv 1218:1219 iv 1218:1219 Here we provide a step-by-step GIG protocol that includes procedures for (i) protein immobilization on an aldehyde-active solid support by reductive amination; (ii) stabilization of fragile sialic acids by carbodiimide coupling; (iii) release of N-glycans by PNGase F digestion; (iv) release of O-glycans by β-elimination using ammonia in the presence of 1-phenyl-3-methyl-5-pyrazolone (PMP) to prevent alditol peeling from O-glycans; (v) mass spectrometry (MS) analysis; and (vi) data analysis for identification of glycans using in-house developed software (GIG Tool; free to download via http://www.biomarkercenter.org/gigtool). 28518173 6 104 from specimen 869:876 arg1 N-glycans 831:839 N-glycans 831:839 N-glycans 831:839 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 6 104 from specimen 869:876 arg1 analysis 819:826 quantitative analysis 806:826 quantitative analysis of N-glycans and O-glycans from a single specimen 806:876 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 6 104 from specimen 869:876 arg1 O-glycans 845:853 O-glycans 845:853 O-glycans 845:853 GIG enables quantitative analysis of N-glycans and O-glycans from a single specimen and can be applied to a high-throughput automated platform. 28518173 9 105 theme ∼3 1922:1923 arg1 d 1925:1925 ∼3 d 1922:1925 ∼3 d 1922:1925 The GIG protocol takes ∼3 d. 28518173 1 106 theme biological 131:140 arg1 activities 142:151 biological activities 131:151 biological activities 131:151 Glycosylation has a pivotal role in a diverse range of biological activities, modulating the structure and function of proteins. 28518173 4 107 theme chemical 562:569 arg1 reactions 571:579 these enzymatic and chemical reactions 542:579 these enzymatic and chemical reactions 542:579 However, it is challenging to combine these enzymatic and chemical reactions in order to analyze both N- and O-glycans. 28518173 8 108 from masses 1602:1607 arg1 extracts 1583:1590 The GIG tool extracts 1570:1590 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags 1570:1829 The GIG tool extracts precursor masses, oxonium ions and glycan fragments from tandem (liquid chromatography (LC)-MS/MS) mass spectra for glycan identification, and reporter ions from quaternary amine containing isobaric tag for glycan (QUANTITY) isobaric tags are used for quantification of the relative abundance of N-glycans. 28518173 2 109 theme oxygen 291:296 arg1 atom 298:301 the oxygen atom 287:301 the oxygen atom (O-linked) of serine and threonine side chains 287:348 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 28518173 3 110 dep released 420:427 arg1 whereas 446:452 whereas 446:452 whereas 446:452 N-glycans can be released by glycosidases, whereas O-glycans are often cleaved by chemical reaction. 28518173 2 111 theme serine 317:322 arg1 atom 298:301 the oxygen atom 287:301 the oxygen atom (O-linked) of serine and threonine side chains 287:348 Glycogens coupled to the nitrogen atom (N-linked) of asparagine side chains or to the oxygen atom (O-linked) of serine and threonine side chains represent the two major protein glycosylation forms. 26240167 4 0 theme dynamics 541:548 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 0 theme dynamics 541:548 arg1 analysis 573:580 mutational analysis 562:580 mutational analysis 562:580 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 0 theme dynamics 541:548 arg1 studies 591:597 kinetic studies 583:597 kinetic studies of in vitro processing events and glycan flux analysis 583:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 1 theme mutational 562:571 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 1 theme mutational 562:571 arg1 analysis 573:580 mutational analysis 562:580 mutational analysis 562:580 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 1 2 theme diverse 158:164 arg1 structures 173:182 diverse glycan structures 158:182 diverse glycan structures 158:182 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 4 3 theme events 622:627 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 3 theme events 622:627 arg1 studies 591:597 kinetic studies 583:597 kinetic studies of in vitro processing events and glycan flux analysis 583:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 0 4 from Analysis 0:7 arg1 reticulum 65:73 the endoplasmic reticulum 49:73 the endoplasmic reticulum 49:73 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 0 4 from Analysis 0:7 arg1 Golgi 83:87 the Golgi 79:87 the Golgi 79:87 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 4 5 theme N-glycan 700:707 arg1 processing 709:718 N-glycan processing 700:718 N-glycan processing 700:718 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 6 theme flux 640:643 arg1 analysis 645:652 glycan flux analysis 633:652 glycan flux analysis 633:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 7 theme glycan 633:638 arg1 analysis 645:652 glycan flux analysis 633:652 glycan flux analysis 633:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 1 8 from generation 144:153 arg1 pathway 201:207 the secretory pathway 187:207 the secretory pathway 187:207 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 1 9 theme glycan 166:171 arg1 structures 173:182 diverse glycan structures 158:182 diverse glycan structures 158:182 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 4 10 theme Molecular 531:539 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 10 theme Molecular 531:539 arg1 analysis 573:580 mutational analysis 562:580 mutational analysis 562:580 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 10 theme Molecular 531:539 arg1 studies 591:597 kinetic studies 583:597 kinetic studies of in vitro processing events and glycan flux analysis 583:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 2 11 theme endoplasmic 279:289 arg1 reticulum 291:299 the endoplasmic reticulum 275:299 the endoplasmic reticulum 275:299 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 1 12 theme structures 173:182 arg1 generation 144:153 the generation 140:153 the generation of diverse glycan structures in the secretory pathway 140:207 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 1 12 theme structures 173:182 arg1 hallmark 94:101 The hallmark 90:101 The hallmark of N-linked protein glycosylation 90:135 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 3 13 theme spectrometry-based 402:419 arg1 analytics 421:429 newly developed mass spectrometry-based analytics 381:429 newly developed mass spectrometry-based analytics 381:429 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 0 14 from remodeling 35:44 arg1 reticulum 65:73 the endoplasmic reticulum 49:73 the endoplasmic reticulum 49:73 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 0 14 from remodeling 35:44 arg1 Golgi 83:87 the Golgi 79:87 the Golgi 79:87 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 2 15 theme cellular 327:334 arg1 setting 336:342 the cellular setting 323:342 the cellular setting for structural diversity 323:367 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 2 16 from processes 239:247 arg1 reticulum 291:299 the endoplasmic reticulum 275:299 the endoplasmic reticulum 275:299 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 2 16 from processes 239:247 arg1 Golgi 309:313 the Golgi 305:313 the Golgi 305:313 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 4 17 theme in 602:603 arg1 events 622:627 in vitro processing events 602:627 in vitro processing events 602:627 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 0 18 theme N-glycan 26:33 arg1 remodeling 35:44 site-specific N-glycan remodeling 12:44 site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi 12:87 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 1 19 theme secretory 191:199 arg1 pathway 201:207 the secretory pathway 187:207 the secretory pathway 187:207 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 0 20 theme site-specific 12:24 arg1 remodeling 35:44 site-specific N-glycan remodeling 12:44 site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi 12:87 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 4 21 theme processing 611:620 arg1 events 622:627 in vitro processing events 602:627 in vitro processing events 602:627 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 0 22 from Golgi 83:87 arg1 Analysis 0:7 Analysis 0:7 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 0:88 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 3 23 theme site-specific 443:455 arg1 remodeling 466:475 site-specific N-glycan remodeling 443:475 site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells 443:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 2 24 theme structural 348:357 arg1 diversity 359:367 structural diversity 348:367 structural diversity 348:367 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 4 25 theme protein 689:695 arg1 role 677:680 the defining role 664:680 the defining role of the protein in N-glycan processing 664:718 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 26 from role 677:680 arg1 processing 709:718 N-glycan processing 700:718 N-glycan processing 700:718 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 3 27 theme protein 490:496 arg1 Pdi1p 498:502 the model protein Pdi1p 480:502 the model protein Pdi1p expressed in insect cells 480:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 0 28 theme remodeling 35:44 arg1 Analysis 0:7 Analysis 0:7 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 0:88 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 3 29 theme N-glycan 457:464 arg1 remodeling 466:475 site-specific N-glycan remodeling 443:475 site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells 443:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 3 30 theme developed 387:395 arg1 analytics 421:429 newly developed mass spectrometry-based analytics 381:429 newly developed mass spectrometry-based analytics 381:429 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 3 31 theme Pdi1p 498:502 arg1 remodeling 466:475 site-specific N-glycan remodeling 443:475 site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells 443:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 0 32 theme endoplasmic 53:63 arg1 reticulum 65:73 the endoplasmic reticulum 49:73 the endoplasmic reticulum 49:73 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 0 33 from reticulum 65:73 arg1 Analysis 0:7 Analysis 0:7 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 0:88 Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi. 26240167 4 34 dep in 602:603 arg1 vitro 605:609 vitro 605:609 vitro 605:609 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 1 35 theme N-linked 106:113 arg1 glycosylation 123:135 N-linked protein glycosylation 106:135 N-linked protein glycosylation 106:135 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 4 36 theme analysis 645:652 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 36 theme analysis 645:652 arg1 studies 591:597 kinetic studies 583:597 kinetic studies of in vitro processing events and glycan flux analysis 583:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 2 37 theme Dynamic 210:216 arg1 processes 239:247 Dynamic, non-template-driven processes 210:247 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi 210:313 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 1 38 theme protein 115:121 arg1 glycosylation 123:135 N-linked protein glycosylation 106:135 N-linked protein glycosylation 106:135 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 4 39 theme kinetic 583:589 arg1 simulation 550:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation 531:559 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 4 39 theme kinetic 583:589 arg1 studies 591:597 kinetic studies 583:597 kinetic studies of in vitro processing events and glycan flux analysis 583:652 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 1 40 link N-linked 106:113 arg1 glycosylation 123:135 N-linked protein glycosylation 106:135 N-linked protein glycosylation 106:135 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 1 41 theme glycosylation 123:135 arg1 generation 144:153 the generation 140:153 the generation of diverse glycan structures in the secretory pathway 140:207 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 1 41 theme glycosylation 123:135 arg1 hallmark 94:101 The hallmark 90:101 The hallmark of N-linked protein glycosylation 90:135 The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. 26240167 3 42 theme insect 517:522 arg1 cells 524:528 insect cells 517:528 insect cells 517:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 2 43 theme remodeling 261:270 arg1 processes 239:247 Dynamic, non-template-driven processes 210:247 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi 210:313 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 4 44 theme defining 668:675 arg1 role 677:680 the defining role 664:680 the defining role of the protein in N-glycan processing 664:718 Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing. 26240167 3 45 theme model 484:488 arg1 Pdi1p 498:502 the model protein Pdi1p 480:502 the model protein Pdi1p expressed in insect cells 480:528 We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. 26240167 2 46 dep Dynamic 210:216 arg1 non-template-driven 219:237 non-template-driven 219:237 non-template-driven 219:237 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 26240167 2 47 theme N-glycan 252:259 arg1 remodeling 261:270 N-glycan remodeling 252:270 N-glycan remodeling 252:270 Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. 27439567 1 0 gly glycosites 210:219 arg2 glycosites 210:219 core-fucosylated versus antennary-fucosylated glycosites 164:219 core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 164:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 1 1 from glycosites 210:219 arg1 glycoproteins 224:236 glycoproteins 224:236 glycoproteins 224:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 1 2 theme core-fucosylated 164:179 arg1 glycosites 210:219 core-fucosylated versus antennary-fucosylated glycosites 164:219 core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 164:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 7 3 theme mass 1190:1193 arg1 profile 1195:1201 a mass profile 1188:1201 a mass profile scan 1188:1206 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 4 4 theme HILIC 600:604 arg1 separation 654:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation 588:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 1 5 gly antennary-fucosylated 188:208 arg1 glycosites 210:219 core-fucosylated versus antennary-fucosylated glycosites 164:219 core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 164:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 7 6 theme sites 1079:1083 arg1 identification 1047:1060 site-specific identification 1033:1060 site-specific identification of glycosylation sites 1033:1083 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 7 6 theme sites 1079:1083 arg1 ability 1093:1099 the ability 1089:1099 the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan 1089:1206 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 4 7 theme multiple 671:678 arg1 fractions 680:688 multiple fractions 671:688 multiple fractions 671:688 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 3 8 theme intact 412:417 arg1 glycoproteins 419:431 intact glycoproteins 412:431 intact glycoproteins 412:431 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 4 9 theme off-line 591:598 arg1 separation 654:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation 588:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 1 10 gly glycoproteins 224:236 arg1 glycoproteins 224:236 glycoproteins 224:236 glycoproteins 224:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 6 11 theme other 896:900 arg1 half 902:905 the other half 892:905 the other half 892:905 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 8 12 gly glycosites 1281:1290 arg2 glycosites 1281:1290 various glycosites 1273:1290 various glycosites in A1AT 1273:1298 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 7 13 gly glycosylation 1065:1077 arg2 sites 1079:1083 glycosylation sites 1065:1083 glycosylation sites 1065:1083 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 3 14 theme glycoproteins 419:431 arg1 digestion 399:407 digestion 399:407 digestion of intact glycoproteins into glycopeptides 399:450 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 1 15 from analysis 152:159 arg1 glycoproteins 224:236 glycoproteins 224:236 glycoproteins 224:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 0 16 gly fucosylation 69:80 arg1 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 4 17 theme truncated 549:557 arg1 glycans 559:565 truncated glycans 549:565 truncated glycans 549:565 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 7 18 theme glycosylation 1065:1077 arg1 sites 1079:1083 glycosylation sites 1065:1083 glycosylation sites 1065:1083 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 3 19 with digestion 472:480 arg1 galactosidase 501:513 galactosidase 501:513 galactosidase 501:513 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 3 19 with digestion 472:480 arg1 sialidase 487:495 sialidase 487:495 sialidase 487:495 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 1 20 theme antennary-fucosylated 188:208 arg1 glycosites 210:219 core-fucosylated versus antennary-fucosylated glycosites 164:219 core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 164:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 3 21 theme double 465:470 arg1 digestion 472:480 double digestion 465:480 double digestion with sialidase and galactosidase 465:513 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 7 22 theme site-specific 1033:1045 arg1 identification 1047:1060 site-specific identification 1033:1060 site-specific identification of glycosylation sites 1033:1083 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 1 23 theme glycosites 210:219 arg1 analysis 152:159 analysis 152:159 analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 152:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 0 24 from analysis 20:27 arg1 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 3 25 gly glycopeptides 438:450 arg2 glycopeptides 438:450 glycopeptides 438:450 glycopeptides 438:450 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 2 26 gly antennary-fucosylated 343:363 arg1 glycosites 365:374 antennary-fucosylated glycosites 343:374 antennary-fucosylated glycosites 343:374 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 8 27 attach present 1262:1268 arg2 fucosylation 1233:1244 antennary fucosylation 1223:1244 antennary fucosylation 1223:1244 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 8 27 attach present 1262:1268 arg2 core 1214:1217 core 1214:1217 core 1214:1217 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 8 27 attach present 1262:1268 arg1 glycosites 1281:1290 various glycosites 1273:1290 various glycosites in A1AT 1273:1298 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 7 28 theme double 1167:1172 arg1 digestion 1174:1182 a double digestion 1165:1182 a double digestion 1165:1182 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 5 29 from glycopeptides 736:748 arg1 fraction 758:765 each fraction 753:765 each fraction 753:765 The glycopeptides in each fraction were treated with PNGase F and then divided into halves. 27439567 7 30 theme scan 1203:1206 arg1 profile 1195:1201 a mass profile 1188:1201 a mass profile scan 1188:1206 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 6 31 theme peptide 863:869 arg1 identification 871:884 peptide identification 863:884 peptide identification 863:884 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 2 32 theme antennary-fucosylated 343:363 arg1 glycosites 365:374 antennary-fucosylated glycosites 343:374 antennary-fucosylated glycosites 343:374 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 4 33 theme chromatography 638:651 arg1 separation 654:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation 588:663 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 3 34 gly glycoproteins 419:431 arg1 glycoproteins 419:431 intact glycoproteins 412:431 intact glycoproteins 412:431 The workflow involves digestion of intact glycoproteins into glycopeptides, followed by double digestion with sialidase and galactosidase. 27439567 1 35 theme MS-based 108:115 arg1 methodology 117:127 A MS-based methodology 106:127 A MS-based methodology 106:127 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 8 36 located present 1262:1268 arg2 fucosylation 1233:1244 antennary fucosylation 1223:1244 antennary fucosylation 1223:1244 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 8 36 located present 1262:1268 arg2 core 1214:1217 core 1214:1217 core 1214:1217 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 8 36 located present 1262:1268 arg1 glycosites 1281:1290 various glycosites 1273:1290 various glycosites in A1AT 1273:1298 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 2 37 contain contains 319:326 arg2 glycosites 365:374 antennary-fucosylated glycosites 343:374 antennary-fucosylated glycosites 343:374 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 37 contain contains 319:326 arg1 glycoprotein 272:283 the glycoprotein alpha-1-antitrypsin 268:303 the glycoprotein alpha-1-antitrypsin (A1AT) 268:310 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 37 contain contains 319:326 arg1 A1AT 306:309 A1AT 306:309 A1AT 306:309 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 37 contain contains 319:326 arg2 core- 333:337 core- 333:337 core- 333:337 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 7 38 theme core 1116:1119 arg1 fucosylation 1121:1132 core fucosylation 1116:1132 core fucosylation 1116:1132 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 0 39 from alpha-1-antitrypsin 85:103 arg1 analysis 20:27 the analysis 16:27 the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin 16:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 4 40 theme resulting 520:528 arg1 glycopeptides 530:542 The resulting glycopeptides 516:542 The resulting glycopeptides with truncated glycans 516:565 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 2 41 gly glycoprotein 272:283 arg1 glycoprotein 272:283 the glycoprotein alpha-1-antitrypsin 268:303 the glycoprotein alpha-1-antitrypsin (A1AT) 268:310 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 41 gly glycoprotein 272:283 arg1 A1AT 306:309 A1AT 306:309 A1AT 306:309 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 4 42 theme hydrophilic 607:617 arg1 chromatography 638:651 hydrophilic interaction liquid chromatography 607:651 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 0 43 from fucosylation 69:80 arg1 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 alpha-1-antitrypsin 85:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 5 44 gly glycopeptides 736:748 arg2 glycopeptides 736:748 The glycopeptides 732:748 The glycopeptides in each fraction 732:765 The glycopeptides in each fraction were treated with PNGase F and then divided into halves. 27439567 8 45 theme various 1273:1279 arg1 glycosites 1281:1290 various glycosites 1273:1290 various glycosites in A1AT 1273:1298 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 0 46 theme structure-specific 50:67 arg1 fucosylation 69:80 site-specific and structure-specific fucosylation 32:80 site-specific and structure-specific fucosylation in alpha-1-antitrypsin 32:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 7 47 theme antennary 1138:1146 arg1 fucosylation 1148:1159 antennary fucosylation 1138:1159 antennary fucosylation 1138:1159 This procedure provided site-specific identification of glycosylation sites and the ability to distinguish core fucosylation and antennary fucosylation via a double digestion and a mass profile scan. 27439567 0 48 theme site-specific 32:44 arg1 fucosylation 69:80 site-specific and structure-specific fucosylation 32:80 site-specific and structure-specific fucosylation in alpha-1-antitrypsin 32:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 4 49 theme liquid 631:636 arg1 chromatography 638:651 hydrophilic interaction liquid chromatography 607:651 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 5 50 theme PNGase 785:790 arg1 F 792:792 PNGase F 785:792 PNGase F 785:792 The glycopeptides in each fraction were treated with PNGase F and then divided into halves. 27439567 8 51 from glycosites 1281:1290 arg1 A1AT 1295:1298 A1AT 1295:1298 A1AT 1295:1298 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 6 52 theme sample 840:845 arg1 half 828:831 One half 824:831 One half of the sample 824:845 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 6 53 theme meladrazine 964:974 arg1 reagent 976:982 a meladrazine reagent 962:982 a meladrazine reagent followed by MS analysis 962:1006 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 4 54 theme interaction 619:629 arg1 chromatography 638:651 hydrophilic interaction liquid chromatography 607:651 an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals 588:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 6 55 theme glycan 925:930 arg1 analysis 932:939 glycan analysis 925:939 glycan analysis 925:939 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 8 56 theme antennary 1223:1231 arg1 fucosylation 1233:1244 antennary fucosylation 1223:1244 antennary fucosylation 1223:1244 Both core and antennary fucosylation are shown to be present on various glycosites in A1AT. 27439567 4 57 theme various 708:714 arg1 intervals 721:729 various time intervals 708:729 various time intervals 708:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 1 58 gly core-fucosylated 164:179 arg1 glycosites 210:219 core-fucosylated versus antennary-fucosylated glycosites 164:219 core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 164:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 4 59 gly glycopeptides 530:542 arg2 glycopeptides 530:542 The resulting glycopeptides 516:542 The resulting glycopeptides with truncated glycans 516:565 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 0 60 theme fucosylation 69:80 arg1 analysis 20:27 the analysis 16:27 the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin 16:103 A procedure for the analysis of site-specific and structure-specific fucosylation in alpha-1-antitrypsin. 27439567 1 61 from glycoproteins 224:236 arg1 analysis 152:159 analysis 152:159 analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins 152:236 A MS-based methodology has been developed for analysis of core-fucosylated versus antennary-fucosylated glycosites in glycoproteins. 27439567 4 62 with glycopeptides 530:542 arg1 glycans 559:565 truncated glycans 549:565 truncated glycans 549:565 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 27439567 6 63 theme MS 996:997 arg1 analysis 999:1006 MS analysis 996:1006 MS analysis 996:1006 One half of the sample was applied for peptide identification while the other half was processed for glycan analysis by derivatizing with a meladrazine reagent followed by MS analysis. 27439567 2 64 gly glycosites 365:374 arg2 glycosites 365:374 antennary-fucosylated glycosites 343:374 antennary-fucosylated glycosites 343:374 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 65 theme alpha-1-antitrypsin 285:303 arg1 glycoprotein 272:283 the glycoprotein alpha-1-antitrypsin 268:303 the glycoprotein alpha-1-antitrypsin (A1AT) 268:310 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 2 65 theme alpha-1-antitrypsin 285:303 arg1 A1AT 306:309 A1AT 306:309 A1AT 306:309 This procedure is applied to the glycoprotein alpha-1-antitrypsin (A1AT), which contains both core- and antennary-fucosylated glycosites. 27439567 4 66 theme time 716:719 arg1 intervals 721:729 various time intervals 708:729 various time intervals 708:729 The resulting glycopeptides with truncated glycans were separated using an off-line HILIC (hydrophilic interaction liquid chromatography) separation where multiple fractions were collected at various time intervals. 25117693 1 0 theme cell-cell 257:265 arg1 interaction 267:277 the receptor-ligand and cell-cell interaction 233:277 interaction 267:277 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 3 1 theme interaction 702:712 arg1 ZIC-HILIC 737:745 ZIC-HILIC 737:745 ZIC-HILIC 737:745 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 3 1 theme interaction 702:712 arg1 chromatography 721:734 Zwitterionic hydrophilic interaction liquid chromatography 677:734 Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) 677:746 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 6 2 theme secreted 1194:1201 arg1 proteins 1203:1210 the secreted proteins 1190:1210 the secreted proteins from A. niger 1190:1224 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 2 3 theme magnetic 541:548 arg1 NMR 561:563 NMR 561:563 NMR 561:563 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 3 theme magnetic 541:548 arg1 resonance 550:558 nuclear magnetic resonance 533:558 nuclear magnetic resonance (NMR) detection 533:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 1 4 theme various 193:199 arg1 interaction 267:277 the receptor-ligand and cell-cell interaction 233:277 interaction 267:277 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 4 theme various 193:199 arg1 processes 212:220 various biological processes 193:220 various biological processes 193:220 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 7 5 theme downstream 1567:1576 arg1 analysis 1582:1589 the downstream NMR analysis 1563:1589 the downstream NMR analysis 1563:1589 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 1 6 theme important 117:125 arg1 modification 158:169 an important and complex post-translational modification 114:169 an important and complex post-translational modification (PTM) 114:175 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 6 theme important 117:125 arg1 glycosylation 99:111 Protein glycosylation 91:111 Protein glycosylation 91:111 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 6 theme important 117:125 arg1 PTM 172:174 PTM 172:174 PTM 172:174 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 6 7 theme NMR 1164:1166 arg1 spectra 1168:1174 The NMR spectra 1160:1174 The NMR spectra 1160:1174 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 3 8 theme pronase 793:799 arg1 digests 803:809 the pronase E digests 789:809 the pronase E digests of RNase B 789:820 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 1 9 theme complex 131:137 arg1 modification 158:169 an important and complex post-translational modification 114:169 an important and complex post-translational modification (PTM) 114:175 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 9 theme complex 131:137 arg1 glycosylation 99:111 Protein glycosylation 91:111 Protein glycosylation 91:111 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 9 theme complex 131:137 arg1 PTM 172:174 PTM 172:174 PTM 172:174 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 4 10 theme simple 948:953 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 10 theme simple 948:953 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 11 theme fast 956:959 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 11 theme fast 956:959 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 3 12 theme RNase 814:818 arg1 B 820:820 RNase B 814:820 RNase B 814:820 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 0 13 theme NMR 76:78 arg1 detection 80:88 NMR detection 76:88 NMR detection 76:88 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection. 25117693 5 14 theme E 1091:1091 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 14 theme E 1091:1091 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 7 15 theme glycoproteomics 1644:1658 arg1 research 1660:1667 the future NMR-based glycoproteomics research 1623:1667 the future NMR-based glycoproteomics research 1623:1667 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 5 16 theme biological 1052:1061 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 16 theme biological 1052:1061 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 7 17 theme future 1627:1632 arg1 research 1660:1667 the future NMR-based glycoproteomics research 1623:1667 the future NMR-based glycoproteomics research 1623:1667 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 1 18 gly glycosylation 99:111 arg1 interaction 267:277 the receptor-ligand and cell-cell interaction 233:277 interaction 267:277 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 18 gly glycosylation 99:111 arg1 processes 212:220 various biological processes 193:220 various biological processes 193:220 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 5 19 theme sample 1071:1076 arg1 analysis 1038:1045 analysis 1038:1045 analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1038:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 2 20 theme conventional 437:448 arg1 time-consuming 579:592 time-consuming 579:592 time-consuming 579:592 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 20 theme conventional 437:448 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 20 theme conventional 437:448 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 5 21 theme Aspergillus 1141:1151 arg1 niger 1153:1157 the fungus Aspergillus niger 1130:1157 the fungus Aspergillus niger 1130:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 2 22 theme column 499:504 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 22 theme column 499:504 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 23 theme glycan 370:375 arg1 structures 377:386 the glycan structures 366:386 the glycan structures of important biological complex samples 366:426 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 0 24 theme Structural 0:9 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis 0:63 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection. 25117693 1 25 theme Protein 91:97 arg1 modification 158:169 an important and complex post-translational modification 114:169 an important and complex post-translational modification (PTM) 114:175 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 25 theme Protein 91:97 arg1 glycosylation 99:111 Protein glycosylation 91:111 Protein glycosylation 91:111 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 3 26 from digests 803:809 arg1 glycans 776:782 the glycans 772:782 the glycans from the pronase E digests of RNase B 772:820 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 2 27 theme samples 420:426 arg1 structures 377:386 the glycan structures 366:386 the glycan structures of important biological complex samples 366:426 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 5 28 theme fungus 1134:1139 arg1 niger 1153:1157 the fungus Aspergillus niger 1130:1157 the fungus Aspergillus niger 1130:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 7 29 theme ZIC-HILIC 1462:1470 arg1 separation 1472:1481 ZIC-HILIC separation 1462:1481 ZIC-HILIC separation coupled with dialysis 1462:1503 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 2 30 theme biological 401:410 arg1 samples 420:426 important biological complex samples 391:426 important biological complex samples 391:426 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 5 31 from digest 1093:1098 arg1 niger 1153:1157 the fungus Aspergillus niger 1130:1157 the fungus Aspergillus niger 1130:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 2 32 dep separation 512:521 arg1 i.e. 478:481 i.e. 478:481 i.e. 478:481 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 33 theme prior 524:528 arg1 time-consuming 579:592 time-consuming 579:592 time-consuming 579:592 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 33 theme prior 524:528 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 33 theme prior 524:528 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 4 34 theme sample 976:981 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 34 theme sample 976:981 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 5 35 theme secreted 1107:1114 arg1 proteins 1116:1123 the secreted proteins 1103:1123 the secreted proteins from the fungus Aspergillus niger 1103:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 3 36 theme glycan 637:642 arg1 strategy 655:662 a glycan enrichment strategy 635:662 a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate 635:868 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 2 37 theme enrichment 457:466 arg1 time-consuming 579:592 time-consuming 579:592 time-consuming 579:592 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 37 theme enrichment 457:466 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 37 theme enrichment 457:466 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 3 38 theme hydrophilic 690:700 arg1 ZIC-HILIC 737:745 ZIC-HILIC 737:745 ZIC-HILIC 737:745 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 3 38 theme hydrophilic 690:700 arg1 chromatography 721:734 Zwitterionic hydrophilic interaction liquid chromatography 677:734 Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) 677:746 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 6 39 theme N-linked 1239:1246 arg1 glycans 1248:1254 both N-linked glycans 1234:1254 both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B 1234:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 40 theme 1→6 1396:1398 arg1 linkages 1400:1407 1→6 linkages 1396:1407 1→6 linkages 1396:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 2 41 theme glycan 450:455 arg1 time-consuming 579:592 time-consuming 579:592 time-consuming 579:592 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 41 theme glycan 450:455 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 41 theme glycan 450:455 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 6 42 link N-linked 1239:1246 arg1 glycans 1248:1254 both N-linked glycans 1234:1254 both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B 1234:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 1 43 theme many 308:311 arg1 functions 324:332 many biological functions 308:332 many biological functions 308:332 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 3 44 theme liquid 714:719 arg1 ZIC-HILIC 737:745 ZIC-HILIC 737:745 ZIC-HILIC 737:745 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 3 44 theme liquid 714:719 arg1 chromatography 721:734 Zwitterionic hydrophilic interaction liquid chromatography 677:734 Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) 677:746 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 7 45 theme NMR 1578:1580 arg1 analysis 1582:1589 the downstream NMR analysis 1563:1589 the downstream NMR analysis 1563:1589 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 6 46 with glycans 1248:1254 arg1 core 1276:1279 a high-mannose core 1261:1279 a high-mannose core similar to the structure of the glycan from RNase B 1261:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 2 47 theme nuclear 533:539 arg1 NMR 561:563 NMR 561:563 NMR 561:563 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 47 theme nuclear 533:539 arg1 resonance 550:558 nuclear magnetic resonance 533:558 nuclear magnetic resonance (NMR) detection 533:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 5 48 from niger 1153:1157 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 48 from niger 1153:1157 arg1 proteins 1116:1123 the secreted proteins 1103:1123 the secreted proteins from the fungus Aspergillus niger 1103:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 48 from niger 1153:1157 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 1 49 theme biological 201:210 arg1 interaction 267:277 the receptor-ligand and cell-cell interaction 233:277 interaction 267:277 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 49 theme biological 201:210 arg1 processes 212:220 various biological processes 193:220 various biological processes 193:220 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 4 50 theme ZIC-HILIC 900:908 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 50 theme ZIC-HILIC 900:908 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 6 51 theme similar 1281:1287 arg1 core 1276:1279 a high-mannose core 1261:1279 a high-mannose core similar to the structure of the glycan from RNase B 1261:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 7 52 theme NMR-based 1634:1642 arg1 research 1660:1667 the future NMR-based glycoproteomics research 1623:1667 the future NMR-based glycoproteomics research 1623:1667 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 1 53 theme biological 313:322 arg1 functions 324:332 many biological functions 308:332 many biological functions 308:332 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 6 54 theme high-mannose 1263:1274 arg1 core 1276:1279 a high-mannose core 1261:1279 a high-mannose core similar to the structure of the glycan from RNase B 1261:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 3 55 theme NMR 835:837 arg1 analysis 839:846 NMR analysis 835:846 NMR analysis of the glycoconjugate 835:868 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 2 56 theme resonance 550:558 arg1 detection 566:574 nuclear magnetic resonance (NMR) detection 533:574 nuclear magnetic resonance (NMR) detection 533:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 6 57 link O-linked 1338:1345 arg1 glycans 1347:1353 O-linked glycans 1338:1353 O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages 1338:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 3 58 theme E 801:801 arg1 digests 803:809 the pronase E digests 789:809 the pronase E digests of RNase B 789:820 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 1 59 theme post-translational 139:156 arg1 modification 158:169 an important and complex post-translational modification 114:169 an important and complex post-translational modification (PTM) 114:175 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 59 theme post-translational 139:156 arg1 glycosylation 99:111 Protein glycosylation 91:111 Protein glycosylation 91:111 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 1 59 theme post-translational 139:156 arg1 PTM 172:174 PTM 172:174 PTM 172:174 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 2 60 theme size-exclusion 484:497 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 60 theme size-exclusion 484:497 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 6 61 theme RNase 1325:1329 arg1 B 1331:1331 RNase B 1325:1331 RNase B 1325:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 5 62 theme pronase 1083:1089 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 62 theme pronase 1083:1089 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 3 63 theme glycoconjugate 855:868 arg1 analysis 839:846 NMR analysis 835:846 NMR analysis of the glycoconjugate 835:868 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 3 64 theme B 820:820 arg1 digests 803:809 the pronase E digests 789:809 the pronase E digests of RNase B 789:820 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 6 65 theme glycan 1313:1318 arg1 structure 1296:1304 the structure 1292:1304 the structure of the glycan from RNase B 1292:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 66 from B 1331:1331 arg1 structure 1296:1304 the structure 1292:1304 the structure of the glycan from RNase B 1292:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 66 from B 1331:1331 arg1 glycan 1313:1318 the glycan 1309:1318 the glycan from RNase B 1309:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 67 with mannose 1363:1369 arg1 1→3 1388:1390 1→3 1388:1390 1→3 1388:1390 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 67 with mannose 1363:1369 arg1 linkages 1400:1407 1→6 linkages 1396:1407 1→6 linkages 1396:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 5 68 theme complex 1063:1069 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 68 theme complex 1063:1069 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 6 69 with glucose 1375:1381 arg1 1→3 1388:1390 1→3 1388:1390 1→3 1388:1390 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 69 with glucose 1375:1381 arg1 linkages 1400:1407 1→6 linkages 1396:1407 1→6 linkages 1396:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 70 contain contain 1226:1232 arg2 glycans 1248:1254 both N-linked glycans 1234:1254 both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B 1234:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 70 contain contain 1226:1232 arg1 proteins 1203:1210 the secreted proteins 1190:1210 the secreted proteins from A. niger 1190:1224 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 6 70 contain contain 1226:1232 arg2 glycans 1347:1353 O-linked glycans 1338:1353 O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages 1338:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 2 71 theme [SEC 506:509 arg1 strategy 468:475 the conventional glycan enrichment strategy 433:475 the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection 433:574 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 2 71 theme [SEC 506:509 arg1 separation 512:521 size-exclusion column [SEC] separation 484:521 size-exclusion column [SEC] separation 484:521 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 6 72 from structure 1296:1304 arg1 B 1331:1331 RNase B 1325:1331 RNase B 1325:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 4 73 theme preparation 983:993 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 73 theme preparation 983:993 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 0 74 theme N- 23:24 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis 0:63 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection. 25117693 6 75 with glycans 1347:1353 arg1 core 1276:1279 a high-mannose core 1261:1279 a high-mannose core similar to the structure of the glycan from RNase B 1261:1331 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 2 76 theme complex 412:418 arg1 samples 420:426 important biological complex samples 391:426 important biological complex samples 391:426 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 0 77 theme O-glycans 30:38 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis 0:63 Structural analysis of N- and O-glycans using ZIC-HILIC/dialysis coupled to NMR detection. 25117693 1 78 theme crucial 292:298 arg1 role 300:303 a crucial role 290:303 a crucial role 290:303 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 25117693 2 79 theme important 391:399 arg1 samples 420:426 important biological complex samples 391:426 important biological complex samples 391:426 However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. 25117693 6 80 theme O-linked 1338:1345 arg1 glycans 1347:1353 O-linked glycans 1338:1353 O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages 1338:1407 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 4 81 theme efficient 966:974 arg1 approach 995:1002 a simple, fast, and efficient sample preparation approach 946:1002 a simple, fast, and efficient sample preparation approach 946:1002 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 4 81 theme efficient 966:974 arg1 enrichment 910:919 the ZIC-HILIC enrichment 896:919 the ZIC-HILIC enrichment coupled with dialysis 896:941 Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is a simple, fast, and efficient sample preparation approach. 25117693 3 82 theme enrichment 644:653 arg1 strategy 655:662 a glycan enrichment strategy 635:662 a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate 635:868 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 5 83 theme proteins 1116:1123 arg1 sample 1071:1076 a biological complex sample 1050:1076 a biological complex sample 1050:1076 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 5 83 theme proteins 1116:1123 arg1 digest 1093:1098 the pronase E digest 1079:1098 the pronase E digest of the secreted proteins from the fungus Aspergillus niger 1079:1157 The approach was thus applied to analysis of a biological complex sample, the pronase E digest of the secreted proteins from the fungus Aspergillus niger. 25117693 7 84 theme compelling 1437:1446 arg1 evidence 1448:1455 compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research 1437:1667 compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research 1437:1667 In all, our study provides compelling evidence that ZIC-HILIC separation coupled with dialysis is very effective and accessible in preparing glycans for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research. 25117693 6 85 from niger 1220:1224 arg1 proteins 1203:1210 the secreted proteins 1190:1210 the secreted proteins from A. niger 1190:1224 The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core similar to the structure of the glycan from RNase B, and O-linked glycans bearing mannose and glucose with 1→3 and 1→6 linkages. 25117693 3 86 theme Zwitterionic 677:688 arg1 ZIC-HILIC 737:745 ZIC-HILIC 737:745 ZIC-HILIC 737:745 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 3 86 theme Zwitterionic 677:688 arg1 chromatography 721:734 Zwitterionic hydrophilic interaction liquid chromatography 677:734 Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) 677:746 In this study, we developed a glycan enrichment strategy that couples Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) with dialysis to enrich the glycans from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. 25117693 1 87 theme receptor-ligand 237:251 arg1 interaction 267:277 the receptor-ligand and cell-cell interaction 233:277 interaction 267:277 Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes, including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. 26618856 0 0 theme Disordered 89:98 arg1 Polypeptides 100:111 Intrinsically Disordered Polypeptides 75:111 Intrinsically Disordered Polypeptides 75:111 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 7 1 theme O-linked 1264:1271 arg1 glycosylation 1273:1285 O-linked glycosylation 1264:1285 O-linked glycosylation 1264:1285 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 7 2 theme binding 1496:1502 arg1 characteristics 1504:1518 the protein binding characteristics 1484:1518 the protein binding characteristics rather than structure 1484:1540 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 1 3 theme modifications 173:185 arg1 one 131:133 one 131:133 one 131:133 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 1 3 theme modifications 173:185 arg1 modifications 173:185 post-translational modifications 154:185 the most common post-translational modifications (PTMs) 138:192 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 1 3 theme modifications 173:185 arg1 PTMs 188:191 PTMs 188:191 PTMs 188:191 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 2 4 from effects 314:320 arg1 function 350:357 protein function 342:357 protein function 342:357 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 5 5 gly glycosylated 904:915 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 5 gly glycosylated 904:915 arg1 tau174-183 965:974 tau174-183 965:974 tau174-183 965:974 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 5 gly glycosylated 904:915 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 5 gly glycosylated 904:915 arg1 forms 936:940 glycosylated and unglycosylated forms 904:940 glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), 904:1019 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 6 theme different 949:957 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 6 theme different 949:957 arg1 tau174-183 965:974 tau174-183 965:974 tau174-183 965:974 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 6 theme different 949:957 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 7 gly unglycosylated 921:934 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 7 gly unglycosylated 921:934 arg1 tau174-183 965:974 tau174-183 965:974 tau174-183 965:974 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 7 gly unglycosylated 921:934 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 7 gly unglycosylated 921:934 arg1 forms 936:940 glycosylated and unglycosylated forms 904:940 glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), 904:1019 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 7 8 theme structural 1327:1336 arg1 ensembles 1338:1346 equilibrium structural ensembles 1315:1346 equilibrium structural ensembles of IDPs, for the cases studied here 1315:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 3 9 theme disordered 500:509 arg1 IDPs 521:524 IDPs 521:524 IDPs 521:524 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 9 theme disordered 500:509 arg1 proteins 511:518 Intrinsically disordered proteins 486:518 Intrinsically disordered proteins (IDPs) 486:525 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 0 10 from Effect 0:5 arg1 Ensemble 63:70 the Equilibrium Structural Ensemble 36:70 the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides 36:111 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 5 11 link O-linked 864:871 arg1 glycosylation 873:885 O-linked glycosylation 864:885 O-linked glycosylation 864:885 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 7 12 from effect 1305:1310 arg1 ensembles 1338:1346 equilibrium structural ensembles 1315:1346 equilibrium structural ensembles of IDPs, for the cases studied here 1315:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 2 13 theme remarkable 303:312 arg1 effects 314:320 remarkable effects 303:320 remarkable effects of glycosylation on protein function 303:357 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 5 14 theme explicit 1033:1040 arg1 simulations 1050:1060 all-atom explicit solvent simulations 1024:1060 all-atom explicit solvent simulations 1024:1060 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 0 15 theme Polypeptides 100:111 arg1 Ensemble 63:70 the Equilibrium Structural Ensemble 36:70 the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides 36:111 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 7 16 theme characteristics 1504:1518 arg1 modulation 1470:1479 modulation 1470:1479 modulation of the protein binding characteristics rather than structure 1470:1540 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 3 17 theme proteins 579:586 arg1 regions 555:561 intrinsically disordered regions 530:561 intrinsically disordered regions (IDRs) 530:568 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 17 theme proteins 579:586 arg1 IDPs 521:524 IDPs 521:524 IDPs 521:524 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 17 theme proteins 579:586 arg1 proteins 511:518 Intrinsically disordered proteins 486:518 Intrinsically disordered proteins (IDPs) 486:525 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 17 theme proteins 579:586 arg1 IDRs 564:567 IDRs 564:567 IDRs 564:567 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 7 18 theme modest 1298:1303 arg1 effect 1305:1310 a modest effect 1296:1310 a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here 1296:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 2 19 theme folding 373:379 arg1 stability 381:389 the folding stability 369:389 the folding stability 369:389 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 2 20 theme binding 461:467 arg1 characteristics 469:483 their binding characteristics 455:483 their binding characteristics 455:483 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 5 21 theme human 980:984 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 21 theme human 980:984 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 21 theme human 980:984 arg1 hIAPP 1013:1017 hIAPP 1013:1017 hIAPP 1013:1017 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 4 22 theme IDP 750:752 arg1 structure 754:762 IDP structure 750:762 IDP structure 750:762 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 6 23 theme aqueous 1089:1095 arg1 solution 1097:1104 aqueous solution 1089:1104 aqueous solution 1089:1104 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 6 24 theme O-linked 1110:1117 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 4 25 theme important 696:704 arg1 question 711:718 An important open question 693:718 An important open question 693:718 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 7 26 theme protein 1488:1494 arg1 characteristics 1504:1518 the protein binding characteristics 1484:1518 the protein binding characteristics rather than structure 1484:1540 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 7 27 link O-linked 1264:1271 arg1 glycosylation 1273:1285 O-linked glycosylation 1264:1285 O-linked glycosylation 1264:1285 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 2 28 theme proteins 281:288 arg1 glycosylation 255:267 glycosylation 255:267 glycosylation of globular proteins 255:288 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 2 29 from work 247:250 arg1 glycosylation 255:267 glycosylation 255:267 glycosylation of globular proteins 255:288 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 6 30 theme glycosylated 1119:1130 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 6 31 theme glycan 1213:1218 arg1 parameters 1220:1229 glycan parameters 1213:1229 glycan parameters 1213:1229 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 7 32 theme IDPs 1351:1354 arg1 ensembles 1338:1346 equilibrium structural ensembles 1315:1346 equilibrium structural ensembles of IDPs, for the cases studied here 1315:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 6 33 from IDPs 1081:1084 arg1 solution 1097:1104 aqueous solution 1089:1104 aqueous solution 1089:1104 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 5 34 theme islet 986:990 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 34 theme islet 986:990 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 34 theme islet 986:990 arg1 hIAPP 1013:1017 hIAPP 1013:1017 hIAPP 1013:1017 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 6 35 theme unglycosylated 1136:1149 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 5 36 theme unglycosylated 921:934 arg1 forms 936:940 glycosylated and unglycosylated forms 904:940 glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), 904:1019 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 37 theme structural 842:851 arg1 effects 853:859 the structural effects 838:859 the structural effects of O-linked glycosylation 838:885 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 2 38 theme glycosylation 325:337 arg1 effects 314:320 remarkable effects 303:320 remarkable effects of glycosylation on protein function 303:357 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 4 39 theme open 706:709 arg1 question 711:718 An important open question 693:718 An important open question 693:718 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 0 40 theme Glycosylation 19:31 arg1 Effect 0:5 Effect 0:5 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 0:112 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 4 41 theme binding 767:773 arg1 characteristics 775:789 binding characteristics 767:789 binding characteristics 767:789 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 5 42 theme O-linked 864:871 arg1 glycosylation 873:885 O-linked glycosylation 864:885 O-linked glycosylation 864:885 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 2 43 gly glycosylation 255:267 arg1 proteins 281:288 globular proteins 272:288 globular proteins 272:288 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 5 44 theme amyloid 992:998 arg1 polypeptide 1000:1010 human islet amyloid polypeptide 980:1010 human islet amyloid polypeptide (hIAPP) 980:1018 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 44 theme amyloid 992:998 arg1 IDPs 959:962 two different IDPs 945:962 two different IDPs 945:962 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 44 theme amyloid 992:998 arg1 hIAPP 1013:1017 hIAPP 1013:1017 hIAPP 1013:1017 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 45 theme IDPs 959:962 arg1 forms 936:940 glycosylated and unglycosylated forms 904:940 glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), 904:1019 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 5 46 theme glycosylation 873:885 arg1 effects 853:859 the structural effects 838:859 the structural effects of O-linked glycosylation 838:885 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 3 47 theme disordered 544:553 arg1 regions 555:561 intrinsically disordered regions 530:561 intrinsically disordered regions (IDRs) 530:568 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 47 theme disordered 544:553 arg1 IDRs 564:567 IDRs 564:567 IDRs 564:567 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 6 48 theme all-atom 1181:1188 arg1 fields 1196:1201 two modern all-atom force fields 1170:1201 two modern all-atom force fields for which glycan parameters are also available 1170:1248 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 3 49 dep function 658:665 arg1 remains 667:673 remains 667:673 remains to be elucidated 667:690 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 1 50 theme large 212:216 arg1 diversity 227:235 a large proteome diversity 210:235 a large proteome diversity 210:235 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 5 51 theme glycosylated 904:915 arg1 forms 936:940 glycosylated and unglycosylated forms 904:940 glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), 904:1019 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 7 52 dep has 1292:1294 arg1 suggests 1391:1398 suggests 1391:1398 suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure 1391:1540 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 2 53 theme Previous 238:245 arg1 work 247:250 Previous work 238:250 Previous work on glycosylation of globular proteins 238:288 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 1 54 theme proteome 218:225 arg1 diversity 227:235 a large proteome diversity 210:235 a large proteome diversity 210:235 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 0 55 theme Structural 52:61 arg1 Ensemble 63:70 the Equilibrium Structural Ensemble 36:70 the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides 36:111 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 6 56 gly glycosylated 1119:1130 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 7 57 theme functional 1409:1418 arg1 role 1420:1423 the functional role 1405:1423 the functional role of glycosylation 1405:1440 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 3 58 gly glycosylated 608:619 arg1 regions 555:561 intrinsically disordered regions 530:561 intrinsically disordered regions (IDRs) 530:568 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 58 gly glycosylated 608:619 arg1 IDPs 521:524 IDPs 521:524 IDPs 521:524 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 58 gly glycosylated 608:619 arg1 proteins 511:518 Intrinsically disordered proteins 486:518 Intrinsically disordered proteins (IDPs) 486:525 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 3 58 gly glycosylated 608:619 arg1 IDRs 564:567 IDRs 564:567 IDRs 564:567 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 0 59 theme Equilibrium 40:50 arg1 Ensemble 63:70 the Equilibrium Structural Ensemble 36:70 the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides 36:111 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 6 60 gly unglycosylated 1136:1149 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 7 61 theme structure 1532:1540 arg1 modulation 1470:1479 modulation 1470:1479 modulation of the protein binding characteristics rather than structure 1470:1540 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 5 62 theme all-atom 1024:1031 arg1 simulations 1050:1060 all-atom explicit solvent simulations 1024:1060 all-atom explicit solvent simulations 1024:1060 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 2 63 theme globular 272:279 arg1 proteins 281:288 globular proteins 272:288 globular proteins 272:288 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 7 64 theme glycosylation 1428:1440 arg1 role 1420:1423 the functional role 1405:1423 the functional role of glycosylation 1405:1440 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 6 65 theme modern 1174:1179 arg1 fields 1196:1201 two modern all-atom force fields 1170:1201 two modern all-atom force fields for which glycan parameters are also available 1170:1248 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 4 66 gly glycosylation 729:741 arg1 IDP 750:752 IDP structure 750:762 IDP structure 750:762 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 4 66 gly glycosylation 729:741 arg1 binding 767:773 binding characteristics 767:789 binding characteristics 767:789 An important open question is, does glycosylation affect IDP structure or binding characteristics or both? 26618856 3 67 theme large 573:577 arg1 proteins 579:586 large proteins 573:586 large proteins 573:586 Intrinsically disordered proteins (IDPs) or intrinsically disordered regions (IDRs) of large proteins are also frequently glycosylated, yet how glycosylation affects their function remains to be elucidated. 26618856 2 68 theme protein 342:348 arg1 function 350:357 protein function 342:357 protein function 342:357 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 6 69 theme force 1190:1194 arg1 fields 1196:1201 two modern all-atom force fields 1170:1201 two modern all-atom force fields for which glycan parameters are also available 1170:1248 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 7 70 theme equilibrium 1315:1325 arg1 ensembles 1338:1346 equilibrium structural ensembles 1315:1346 equilibrium structural ensembles of IDPs, for the cases studied here 1315:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 2 71 gly glycosylation 325:337 arg1 protein 342:348 protein function 342:357 protein function 342:357 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 2 72 theme protein 425:431 arg1 surface 433:439 the protein surface 421:439 the protein surface which affects their binding characteristics 421:483 Previous work on glycosylation of globular proteins has revealed remarkable effects of glycosylation on protein function, altering the folding stability and structure and/or altering the protein surface which affects their binding characteristics. 26618856 5 73 theme solvent 1042:1048 arg1 simulations 1050:1060 all-atom explicit solvent simulations 1024:1060 all-atom explicit solvent simulations 1024:1060 In this work, we particularly address the structural effects of O-linked glycosylation by investigating glycosylated and unglycosylated forms of two different IDPs, tau174-183 and human islet amyloid polypeptide (hIAPP), by all-atom explicit solvent simulations. 26618856 1 74 theme common 147:152 arg1 PTMs 188:191 PTMs 188:191 PTMs 188:191 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 1 74 theme common 147:152 arg1 modifications 173:185 post-translational modifications 154:185 the most common post-translational modifications (PTMs) 138:192 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 6 75 link O-linked 1110:1117 arg1 forms 1151:1155 O-linked glycosylated and unglycosylated forms 1110:1155 O-linked glycosylated and unglycosylated forms 1110:1155 We simulate these IDPs in aqueous solution for O-linked glycosylated and unglycosylated forms by employing two modern all-atom force fields for which glycan parameters are also available. 26618856 0 76 theme O-Linked 10:17 arg1 Glycosylation 19:31 O-Linked Glycosylation 10:31 O-Linked Glycosylation 10:31 Effect of O-Linked Glycosylation on the Equilibrium Structural Ensemble of Intrinsically Disordered Polypeptides. 26618856 7 77 contain has 1292:1294 arg1 glycosylation 1273:1285 O-linked glycosylation 1264:1285 O-linked glycosylation 1264:1285 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 7 77 contain has 1292:1294 arg2 effect 1305:1310 a modest effect 1296:1310 a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here 1296:1382 We find that O-linked glycosylation only has a modest effect on equilibrium structural ensembles of IDPs, for the cases studied here, which suggests that the functional role of glycosylation may be primarily exerted by modulation of the protein binding characteristics rather than structure. 26618856 1 78 theme post-translational 154:171 arg1 PTMs 188:191 PTMs 188:191 PTMs 188:191 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 26618856 1 78 theme post-translational 154:171 arg1 modifications 173:185 post-translational modifications 154:185 the most common post-translational modifications (PTMs) 138:192 Glycosylation is one of the most common post-translational modifications (PTMs), which provides a large proteome diversity. 28077298 6 0 theme evolutionary 1493:1504 arg1 interest 1506:1513 glycoanalytical and evolutionary interest 1473:1513 glycoanalytical and evolutionary interest 1473:1513 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 3 1 theme LC-MALDI-TOF 583:594 arg1 approach 599:606 an off-line LC-MALDI-TOF MS approach 571:606 an off-line LC-MALDI-TOF MS approach 571:606 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 4 2 theme sulphated 830:838 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 3 theme ni 777:778 arg1 line 795:798 the commonly-used T. ni High Five cell line 756:798 the commonly-used T. ni High Five cell line 756:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 4 theme lepidopteran 648:659 arg1 dispar 734:739 the gypsy moth Lymantria dispar 709:739 the gypsy moth Lymantria dispar 709:739 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 4 theme lepidopteran 648:659 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 4 theme lepidopteran 648:659 arg1 ni 702:703 the cabbage looper Trichoplusia ni 670:703 the cabbage looper Trichoplusia ni 670:703 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 5 theme glycoprotein 1620:1631 arg1 production 1633:1642 recombinant glycoprotein production 1608:1642 recombinant glycoprotein production 1608:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 5 6 theme zwitterionic 1310:1321 arg1 glycans 1323:1329 5% anionic and/or zwitterionic glycans 1292:1329 5% anionic and/or zwitterionic glycans previously not found in these species 1292:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 6 7 from glycans 1426:1432 arg1 data 1453:1456 the Lepidoptera data 1437:1456 the Lepidoptera data 1437:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 5 8 dep CONCLUSION 1161:1170 arg1 analysed 1198:1205 analysed 1198:1205 analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species 1198:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 3 9 theme High 780:783 arg1 line 795:798 the commonly-used T. ni High Five cell line 756:798 the commonly-used T. ni High Five cell line 756:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 4 10 theme antennal 891:898 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 11 theme species 661:667 arg1 N-glycomes 630:639 the N-glycomes 626:639 the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line 626:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 12 theme anionic 1401:1407 arg1 glycans 1426:1432 anionic and zwitterionic glycans 1401:1432 anionic and zwitterionic glycans in the Lepidoptera data 1401:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 6 13 theme glycoanalytical 1473:1487 arg1 interest 1506:1513 glycoanalytical and evolutionary interest 1473:1513 glycoanalytical and evolutionary interest 1473:1513 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 4 14 theme familiar 1020:1027 arg1 modification 997:1008 a modification 995:1008 a modification otherwise familiar from nematodes 995:1042 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 14 theme familiar 1020:1027 arg1 phosphorylcholine 948:964 the zwitterion phosphorylcholine 933:964 the zwitterion phosphorylcholine on antennal GlcNAc residues 933:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 15 theme commonly-used 760:772 arg1 ni 777:778 the commonly-used T. ni 756:778 the commonly-used T. ni High Five cell line 756:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 4 16 theme Lewis-like 880:889 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 17 theme Trichoplusia 689:700 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 17 theme Trichoplusia 689:700 arg1 ni 702:703 the cabbage looper Trichoplusia ni 670:703 the cabbage looper Trichoplusia ni 670:703 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 18 theme potential 1584:1592 arg1 lines 1574:1578 lepidopteran cell lines 1556:1578 lepidopteran cell lines 1556:1578 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 6 18 theme potential 1584:1592 arg1 factories 1594:1602 potential factories 1584:1602 potential factories for recombinant glycoprotein production 1584:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 2 19 theme antibodies 276:285 arg1 binding 241:247 pathogen binding 232:247 pathogen binding 232:247 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 2 19 theme antibodies 276:285 arg1 production 252:261 production 252:261 production of humanised antibodies 252:285 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 5 20 theme anionic 1295:1301 arg1 glycans 1323:1329 5% anionic and/or zwitterionic glycans 1292:1329 5% anionic and/or zwitterionic glycans previously not found in these species 1292:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 3 21 theme T. 774:775 arg1 ni 777:778 the commonly-used T. ni 756:778 the commonly-used T. ni High Five cell line 756:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 22 theme cell 1569:1572 arg1 lines 1574:1578 lepidopteran cell lines 1556:1578 lepidopteran cell lines 1556:1578 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 6 22 theme cell 1569:1572 arg1 factories 1594:1602 potential factories 1584:1602 potential factories for recombinant glycoprotein production 1584:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 2 23 theme humanised 266:274 arg1 antibodies 276:285 humanised antibodies 266:285 humanised antibodies 266:285 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 4 24 theme glycolipid-like 1074:1088 arg1 antennae 1090:1097 glycolipid-like antennae 1074:1097 glycolipid-like antennae containing α-linked N-acetylgalactosamine 1074:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 25 theme fucosylated 900:910 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 26 theme cabbage 674:680 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 26 theme cabbage 674:680 arg1 ni 702:703 the cabbage looper Trichoplusia ni 670:703 the cabbage looper Trichoplusia ni 670:703 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 4 27 from structures 912:921 arg1 residues 985:992 antennal GlcNAc residues 969:992 antennal GlcNAc residues 969:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 28 theme GlcNAc 978:983 arg1 residues 985:992 antennal GlcNAc residues 969:992 antennal GlcNAc residues 969:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 29 theme α-linked 1110:1117 arg1 N-acetylgalactosamine 1119:1139 α-linked N-acetylgalactosamine 1110:1139 α-linked N-acetylgalactosamine 1110:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 30 from phosphorylcholine 948:964 arg1 residues 985:992 antennal GlcNAc residues 969:992 antennal GlcNAc residues 969:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 0 31 theme underestimated 4:17 arg1 N-glycomes 19:28 The underestimated N-glycomes 0:28 The underestimated N-glycomes of lepidopteran species 0:52 The underestimated N-glycomes of lepidopteran species. 28077298 3 32 theme looper 682:687 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 32 theme looper 682:687 arg1 ni 702:703 the cabbage looper Trichoplusia ni 670:703 the cabbage looper Trichoplusia ni 670:703 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 33 dep SIGNIFICANCE 1370:1381 arg1 occurrence 1387:1396 The occurrence 1383:1396 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data 1370:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 6 34 gly glycoprotein 1620:1631 arg1 glycoprotein 1620:1631 recombinant glycoprotein production 1608:1642 recombinant glycoprotein production 1608:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 4 35 with N-glycans 1059:1067 arg1 antennae 1090:1097 glycolipid-like antennae 1074:1097 glycolipid-like antennae containing α-linked N-acetylgalactosamine 1074:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 36 dep species 661:667 arg1 dispar 734:739 the gypsy moth Lymantria dispar 709:739 the gypsy moth Lymantria dispar 709:739 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 36 dep species 661:667 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 36 dep species 661:667 arg1 ni 702:703 the cabbage looper Trichoplusia ni 670:703 the cabbage looper Trichoplusia ni 670:703 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 37 theme gypsy 713:717 arg1 dispar 734:739 the gypsy moth Lymantria dispar 709:739 the gypsy moth Lymantria dispar 709:739 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 37 theme gypsy 713:717 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 38 theme recombinant 1608:1618 arg1 production 1633:1642 recombinant glycoprotein production 1608:1642 recombinant glycoprotein production 1608:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 2 39 theme long 294:297 arg1 time 299:302 a long time 292:302 a long time 292:302 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 6 40 theme biotechnological 1526:1541 arg1 relevance 1543:1551 biotechnological relevance 1526:1551 biotechnological relevance 1526:1551 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 2 41 theme more 405:408 arg1 species 410:416 more species 405:416 more species 405:416 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 3 42 theme moth 719:722 arg1 dispar 734:739 the gypsy moth Lymantria dispar 709:739 the gypsy moth Lymantria dispar 709:739 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 42 theme moth 719:722 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 2 43 theme structural 506:515 arg1 diversity 517:525 a large structural diversity 498:525 a large structural diversity 498:525 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 4 44 gly fucosylated 900:910 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 2 45 theme large 500:504 arg1 diversity 517:525 a large structural diversity 498:525 a large structural diversity 498:525 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 5 46 theme 5 1292:1292 arg1 % 1293:1293 % 1293:1293 % 1293:1293 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 6 47 from data 1453:1456 arg1 occurrence 1387:1396 The occurrence 1383:1396 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data 1370:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 0 48 theme species 46:52 arg1 N-glycomes 19:28 The underestimated N-glycomes 0:28 The underestimated N-glycomes of lepidopteran species 0:52 The underestimated N-glycomes of lepidopteran species. 28077298 5 49 theme lepidopteran 1176:1187 arg1 glycomes 1189:1196 The lepidopteran glycomes 1172:1196 The lepidopteran glycomes 1172:1196 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 6 50 theme Lepidoptera 1441:1451 arg1 data 1453:1456 the Lepidoptera data 1437:1456 the Lepidoptera data 1437:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 5 51 theme % 1293:1293 arg1 glycans 1323:1329 5% anionic and/or zwitterionic glycans 1292:1329 5% anionic and/or zwitterionic glycans previously not found in these species 1292:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 0 52 theme lepidopteran 33:44 arg1 species 46:52 lepidopteran species 33:52 lepidopteran species 33:52 The underestimated N-glycomes of lepidopteran species. 28077298 4 53 theme zwitterion 937:946 arg1 modification 997:1008 a modification 995:1008 a modification otherwise familiar from nematodes 995:1042 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 53 theme zwitterion 937:946 arg1 phosphorylcholine 948:964 the zwitterion phosphorylcholine 933:964 the zwitterion phosphorylcholine on antennal GlcNAc residues 933:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 6 54 theme zwitterionic 1413:1424 arg1 glycans 1426:1432 anionic and zwitterionic glycans 1401:1432 anionic and zwitterionic glycans in the Lepidoptera data 1401:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 4 55 link α-linked 1110:1117 arg1 N-acetylgalactosamine 1119:1139 α-linked N-acetylgalactosamine 1110:1139 α-linked N-acetylgalactosamine 1110:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 5 56 located found 1346:1350 arg2 glycans 1323:1329 5% anionic and/or zwitterionic glycans 1292:1329 5% anionic and/or zwitterionic glycans previously not found in these species 1292:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 5 56 located found 1346:1350 arg1 species 1361:1367 these species 1355:1367 these species 1355:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 4 57 theme antennal 969:976 arg1 residues 985:992 antennal GlcNAc residues 969:992 antennal GlcNAc residues 969:992 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 58 contain containing 1099:1108 arg1 antennae 1090:1097 glycolipid-like antennae 1074:1097 glycolipid-like antennae containing α-linked N-acetylgalactosamine 1074:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 58 contain containing 1099:1108 arg2 N-acetylgalactosamine 1119:1139 α-linked N-acetylgalactosamine 1110:1139 α-linked N-acetylgalactosamine 1110:1139 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 2 59 theme aspects 163:169 arg1 Many 149:152 Many 149:152 Many 149:152 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 2 59 theme aspects 163:169 arg1 aspects 163:169 these aspects 157:169 these aspects 157:169 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 6 60 theme glycans 1426:1432 arg1 occurrence 1387:1396 The occurrence 1383:1396 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data 1370:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 4 61 theme difucosylated 862:874 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 62 theme Lymantria 724:732 arg1 dispar 734:739 the gypsy moth Lymantria dispar 709:739 the gypsy moth Lymantria dispar 709:739 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 3 62 theme Lymantria 724:732 arg1 species 661:667 two lepidopteran species 644:667 two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) 644:740 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 2 63 theme interspecies 531:542 arg1 variability 544:554 interspecies variability 531:554 interspecies variability 531:554 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 6 64 from occurrence 1387:1396 arg1 data 1453:1456 the Lepidoptera data 1437:1456 the Lepidoptera data 1437:1456 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 3 65 theme line 795:798 arg1 N-glycomes 630:639 the N-glycomes 626:639 the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line 626:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 1 66 theme BACKGROUND 55:64 arg1 Insects 66:72 BACKGROUND Insects 55:72 BACKGROUND Insects 55:72 BACKGROUND Insects are significant to the environment, agriculture, health and biotechnology. 28077298 4 67 theme glucuronylated 841:854 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 3 68 theme MS 596:597 arg1 approach 599:606 an off-line LC-MALDI-TOF MS approach 571:606 an off-line LC-MALDI-TOF MS approach 571:606 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 4 69 gly difucosylated 862:874 arg1 structures 912:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures 830:921 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 70 dep RESULTS 801:807 arg1 revealed 1151:1158 revealed 1151:1158 were also revealed 1141:1158 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 70 dep RESULTS 801:807 arg1 detected 812:819 detected 812:819 detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes 812:1042 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 4 71 from nematodes 1034:1042 arg1 familiar 1020:1027 familiar 1020:1027 familiar 1020:1027 RESULTS We detected not only sulphated, glucuronylated, core difucosylated and Lewis-like antennal fucosylated structures, but also the zwitterion phosphorylcholine on antennal GlcNAc residues, a modification otherwise familiar from nematodes; in L. dispar, N-glycans with glycolipid-like antennae containing α-linked N-acetylgalactosamine were also revealed. 28077298 5 72 theme core 1224:1227 arg1 α1,3-fucosylation 1229:1245 core α1,3-fucosylation 1224:1245 core α1,3-fucosylation 1224:1245 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 5 73 dep analysed 1198:1205 arg1 display 1216:1222 display 1216:1222 analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species 1198:1367 CONCLUSION The lepidopteran glycomes analysed not only display core α1,3-fucosylation, which is foreign to mammals, but also up to 5% anionic and/or zwitterionic glycans previously not found in these species. 28077298 3 74 theme off-line 574:581 arg1 approach 599:606 an off-line LC-MALDI-TOF MS approach 571:606 an off-line LC-MALDI-TOF MS approach 571:606 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 6 75 theme lepidopteran 1556:1567 arg1 lines 1574:1578 lepidopteran cell lines 1556:1578 lepidopteran cell lines 1556:1578 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 6 75 theme lepidopteran 1556:1567 arg1 factories 1594:1602 potential factories 1584:1602 potential factories for recombinant glycoprotein production 1584:1642 SIGNIFICANCE The occurrence of anionic and zwitterionic glycans in the Lepidoptera data is not only of glycoanalytical and evolutionary interest, but is of biotechnological relevance as lepidopteran cell lines are potential factories for recombinant glycoprotein production. 28077298 2 76 theme N-glycosylation 340:354 arg1 potentials 356:365 insect N-glycosylation potentials 333:365 insect N-glycosylation potentials 333:365 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 3 77 theme cell 790:793 arg1 line 795:798 the commonly-used T. ni High Five cell line 756:798 the commonly-used T. ni High Five cell line 756:798 METHODS Using an off-line LC-MALDI-TOF MS approach, we have analysed the N-glycomes of two lepidopteran species (the cabbage looper Trichoplusia ni and the gypsy moth Lymantria dispar) as well as of the commonly-used T. ni High Five cell line. 28077298 2 78 theme pathogen 232:239 arg1 binding 241:247 pathogen binding 232:247 pathogen binding 232:247 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 28077298 2 79 theme insect 333:338 arg1 potentials 356:365 insect N-glycosylation potentials 333:365 insect N-glycosylation potentials 333:365 Many of these aspects display some relationship to glycosylation, e.g., in case of pathogen binding or production of humanised antibodies; for a long time, it has been considered that insect N-glycosylation potentials are rather similar and simple, but as more species are glycomically analysed in depth, it is becoming obvious that there is indeed a large structural diversity and interspecies variability. 27928741 7 0 from protein 1256:1262 arg1 glycopeptides 1204:1216 glycopeptides 1204:1216 glycopeptides from the haemagglutinin-neuraminidase protein 1204:1262 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 13 1 theme disease 2071:2077 arg1 virus 2079:2083 Newcastle disease virus 2061:2083 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 10 2 theme hybrid 1563:1568 arg1 glycans 1570:1576 complex or hybrid glycans 1552:1576 glycans 1570:1576 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 1 3 gly glycoproteins 230:242 arg1 glycoproteins 230:242 haemagglutinin-neuraminidase glycoproteins 201:242 haemagglutinin-neuraminidase glycoproteins 201:242 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 1 3 gly glycoproteins 230:242 arg1 proteins 264:271 their attachment proteins 247:271 their attachment proteins 247:271 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 6 4 theme propagated 912:921 arg1 virions 923:929 propagated virions 912:929 propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66 912:982 Haemagglutinin-neuraminidase was derived from egg propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66. 27928741 13 5 theme distribution 1988:1999 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 5 theme distribution 1988:1999 arg1 assessment 2118:2127 assessment 2118:2127 assessment of the functional significance of the O-linked glycan in the stalk domain of this protein 2118:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 5 6 theme glycan 698:703 arg1 compositions 705:716 the glycan compositions 694:716 the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species 694:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 9 7 theme N-linked 1379:1386 arg1 sites 1388:1392 N-linked sites 1379:1392 N-linked sites 433 and 481 1379:1404 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 3 8 link N-linked 438:445 arg1 glycans 447:453 the N-linked glycans 434:453 the N-linked glycans present on these proteins 434:479 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 1 9 theme haemagglutinin-neuraminidase 201:228 arg1 glycoproteins 230:242 haemagglutinin-neuraminidase glycoproteins 201:242 haemagglutinin-neuraminidase glycoproteins 201:242 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 1 9 theme haemagglutinin-neuraminidase 201:228 arg1 proteins 264:271 their attachment proteins 247:271 their attachment proteins 247:271 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 5 10 theme Newcastle 777:785 arg1 virus 795:799 the Avulavirus Newcastle disease virus 762:799 the Avulavirus Newcastle disease virus 762:799 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 13 11 theme future 1944:1949 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 2 12 theme sialidase 381:389 arg1 activity 391:398 haemagglutination and sialidase activity 359:398 haemagglutination and sialidase activity 359:398 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 9 13 located observed 1483:1490 arg1 site 1495:1498 site 481 1495:1502 site 481 1495:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 9 13 located observed 1483:1490 arg2 glycans 1470:1476 paucimannose glycans 1457:1476 paucimannose glycans also observed at site 481 1457:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 13 14 theme glycomic 1964:1971 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 2 15 theme haemagglutination 359:375 arg1 activity 391:398 haemagglutination and sialidase activity 359:398 haemagglutination and sialidase activity 359:398 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 1 16 theme attachment 253:262 arg1 glycoproteins 230:242 haemagglutinin-neuraminidase glycoproteins 201:242 haemagglutinin-neuraminidase glycoproteins 201:242 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 1 16 theme attachment 253:262 arg1 proteins 264:271 their attachment proteins 247:271 their attachment proteins 247:271 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 5 17 theme Newcastle 815:823 arg1 disease 825:831 Newcastle disease 815:831 Newcastle disease 815:831 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 13 18 from studies 1973:1979 arg1 domain 2196:2201 the stalk domain 2186:2201 the stalk domain of this protein 2186:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 18 from studies 1973:1979 arg1 protein 2211:2217 this protein 2206:2217 this protein 2206:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 2 19 gly glycoproteins 280:292 arg1 proteins 335:342 oligomeric type II integral membrane proteins 298:342 oligomeric type II integral membrane proteins 298:342 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 19 gly glycoproteins 280:292 arg1 glycoproteins 280:292 These glycoproteins 274:292 These glycoproteins 274:292 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 7 20 theme haemagglutinin-neuraminidase 1227:1254 arg1 protein 1256:1262 the haemagglutinin-neuraminidase protein 1223:1262 the haemagglutinin-neuraminidase protein 1223:1262 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 1 21 theme genera 145:150 arg1 Members 89:95 Members 89:95 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses 89:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 13 22 theme significance 2147:2158 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 22 theme significance 2147:2158 arg1 assessment 2118:2127 assessment 2118:2127 assessment of the functional significance of the O-linked glycan in the stalk domain of this protein 2118:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 5 23 theme virus 795:799 arg1 haemagglutinin-neuraminidase 730:757 haemagglutinin-neuraminidase 730:757 haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species 730:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 9 24 theme mannose 1436:1442 arg1 glycans 1444:1450 high mannose glycans 1431:1450 high mannose glycans with paucimannose glycans also observed at site 481 1431:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 2 25 theme integral 317:324 arg1 proteins 335:342 oligomeric type II integral membrane proteins 298:342 oligomeric type II integral membrane proteins 298:342 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 25 theme integral 317:324 arg1 glycoproteins 280:292 These glycoproteins 274:292 These glycoproteins 274:292 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 7 26 theme electron-transfer 1140:1156 arg1 dissociation 1158:1169 electron-transfer dissociation 1140:1169 electron-transfer dissociation 1140:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 5 27 theme avian 847:851 arg1 species 853:859 avian species 847:859 avian species 847:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 4 28 gly heterogeneity 601:613 arg1 glycans 624:630 these glycans 618:630 these glycans 618:630 However, site-specific heterogeneity of these glycans has not been defined. 27928741 1 29 theme family 175:180 arg1 genera 145:150 the Avulavirus, Respirovirus and Rubulavirus genera 100:150 the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses 100:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 11 30 theme hybrid 1691:1696 arg1 glycans 1707:1713 complex or hybrid N-linked glycans 1680:1713 glycans 1707:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 13 31 link N-linked 2029:2036 arg1 sites 2052:2056 N-linked glycosylation sites 2029:2056 N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 2029:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 31 link N-linked 2029:2036 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 7 32 theme higher-energy 1099:1111 arg1 dissociation 1123:1134 higher-energy collision dissociation 1099:1134 higher-energy collision dissociation 1099:1134 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 1 33 theme viruses 185:191 arg1 family 175:180 the Paramyxoviridae family 155:180 the Paramyxoviridae family of viruses 155:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 12 34 theme haemagglutinin-neuraminidase 1868:1895 arg1 protein 1897:1903 the haemagglutinin-neuraminidase protein 1864:1903 the haemagglutinin-neuraminidase protein 1864:1903 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 10 35 theme compositions 1595:1606 arg1 many 1583:1586 many 1583:1586 many 1583:1586 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 35 theme compositions 1595:1606 arg1 compositions 1595:1606 the compositions 1591:1606 the compositions containing variations of fucose and sulfate or phosphate 1591:1663 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 36 theme complex 1552:1558 arg1 glycans 1570:1576 complex or hybrid glycans 1552:1576 glycans 1570:1576 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 9 37 contain contain 1423:1429 arg2 glycans 1444:1450 high mannose glycans 1431:1450 high mannose glycans with paucimannose glycans also observed at site 481 1431:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 9 37 contain contain 1423:1429 arg1 sites 1388:1392 N-linked sites 1379:1392 N-linked sites 433 and 481 1379:1404 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 8 38 dep residues 1338:1345 arg1 481 1360:1362 481 1360:1362 481 1360:1362 Overall 63, 58, and 37 glycan compositions were identified at asparagine residues 341, 433 and 481, respectively. 27928741 8 38 dep residues 1338:1345 arg1 433 1352:1354 433 1352:1354 433 1352:1354 Overall 63, 58, and 37 glycan compositions were identified at asparagine residues 341, 433 and 481, respectively. 27928741 8 38 dep residues 1338:1345 arg1 341 1347:1349 341 1347:1349 341 1347:1349 Overall 63, 58, and 37 glycan compositions were identified at asparagine residues 341, 433 and 481, respectively. 27928741 9 39 with glycans 1444:1450 arg1 glycans 1470:1476 paucimannose glycans 1457:1476 paucimannose glycans also observed at site 481 1457:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 7 40 theme collision 1067:1075 arg1 dissociation 1085:1096 collision induced dissociation 1067:1096 collision induced dissociation 1067:1096 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 13 41 from assessment 2118:2127 arg1 domain 2196:2201 the stalk domain 2186:2201 the stalk domain of this protein 2186:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 41 from assessment 2118:2127 arg1 protein 2211:2217 this protein 2206:2217 this protein 2206:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 12 42 theme stalk 1848:1852 arg1 domain 1854:1859 the stalk domain 1844:1859 the stalk domain of the haemagglutinin-neuraminidase protein 1844:1903 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 12 42 theme stalk 1848:1852 arg1 protein 1897:1903 the haemagglutinin-neuraminidase protein 1864:1903 the haemagglutinin-neuraminidase protein 1864:1903 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 10 43 theme fucose 1633:1638 arg1 variations 1619:1628 variations 1619:1628 variations of fucose and sulfate or phosphate 1619:1663 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 13 44 theme haemagglutinin-neuraminidase 2085:2112 arg1 sites 2052:2056 N-linked glycosylation sites 2029:2056 N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 2029:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 44 theme haemagglutinin-neuraminidase 2085:2112 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 0 45 theme Site-specific 0:12 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of the Newcastle disease virus 0:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 12 46 theme O-linked 1802:1809 arg1 glycopeptide 1811:1822 a previously undocumented O-linked glycopeptide 1776:1822 a previously undocumented O-linked glycopeptide 1776:1822 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 13 47 theme glycosylation 2038:2050 arg1 sites 2052:2056 N-linked glycosylation sites 2029:2056 N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 2029:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 47 theme glycosylation 2038:2050 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 3 48 theme host 529:532 arg1 cells 534:538 host cells 529:538 host cells 529:538 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 6 49 theme strain 970:975 arg1 V4-VAR 934:939 V4-VAR 934:939 V4-VAR 934:939 Haemagglutinin-neuraminidase was derived from egg propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66. 27928741 6 49 theme strain 970:975 arg1 isolate 945:951 an isolate 942:951 an isolate of the avirulent strain QLD/66 942:982 Haemagglutinin-neuraminidase was derived from egg propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66. 27928741 13 50 from significance 2147:2158 arg1 domain 2196:2201 the stalk domain 2186:2201 the stalk domain of this protein 2186:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 50 from significance 2147:2158 arg1 protein 2211:2217 this protein 2206:2217 this protein 2206:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 0 51 theme Newcastle 35:43 arg1 virus 53:57 the Newcastle disease virus 31:57 the Newcastle disease virus 31:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 0 52 gly glycosylation 14:26 arg1 virus 53:57 the Newcastle disease virus 31:57 the Newcastle disease virus 31:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 0 53 theme virus 53:57 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of the Newcastle disease virus 0:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 12 54 gly glycopeptide 1811:1822 arg2 glycopeptide 1811:1822 a previously undocumented O-linked glycopeptide 1776:1822 a previously undocumented O-linked glycopeptide 1776:1822 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 13 55 theme O-linked 2167:2174 arg1 glycan 2176:2181 the O-linked glycan 2163:2181 the O-linked glycan 2163:2181 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 11 56 theme N-linked 1698:1705 arg1 glycans 1707:1713 complex or hybrid N-linked glycans 1680:1713 glycans 1707:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 7 57 theme spectrometry 1033:1044 arg1 strategies 1046:1055 Reverse-phase liquid chromatography tandem mass spectrometry strategies 985:1055 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 7 57 theme spectrometry 1033:1044 arg1 dissociation 1158:1169 electron-transfer dissociation 1140:1169 electron-transfer dissociation 1140:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 7 57 theme spectrometry 1033:1044 arg1 dissociation 1123:1134 higher-energy collision dissociation 1099:1134 higher-energy collision dissociation 1099:1134 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 7 57 theme spectrometry 1033:1044 arg1 dissociation 1085:1096 collision induced dissociation 1067:1096 collision induced dissociation 1067:1096 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 10 58 contain contained 1542:1550 arg1 residues 1516:1523 Asparagine residues 341, 433 and 481 1505:1540 Asparagine residues 341, 433 and 481 1505:1540 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 58 contain contained 1542:1550 arg2 glycans 1570:1576 complex or hybrid glycans 1552:1576 glycans 1570:1576 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 7 59 theme tandem 1021:1026 arg1 spectrometry 1033:1044 Reverse-phase liquid chromatography tandem mass spectrometry 985:1044 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 13 60 link O-linked 2167:2174 arg1 glycan 2176:2181 the O-linked glycan 2163:2181 the O-linked glycan 2163:2181 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 61 theme structures 2011:2020 arg1 distribution 1988:1999 the distribution 1984:1999 the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1984:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 3 62 theme immune 563:568 arg1 system 570:575 the host immune system 554:575 the host immune system 554:575 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 7 63 theme liquid 999:1004 arg1 chromatography 1006:1019 Reverse-phase liquid chromatography 985:1019 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 11 64 link N-linked 1698:1705 arg1 glycans 1707:1713 complex or hybrid N-linked glycans 1680:1713 glycans 1707:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 8 65 theme asparagine 1327:1336 arg1 residues 1338:1345 asparagine residues 341, 433 and 481 1327:1362 asparagine residues 341, 433 and 481 1327:1362 Overall 63, 58, and 37 glycan compositions were identified at asparagine residues 341, 433 and 481, respectively. 27928741 12 66 link O-linked 1802:1809 arg1 glycopeptide 1811:1822 a previously undocumented O-linked glycopeptide 1776:1822 a previously undocumented O-linked glycopeptide 1776:1822 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 13 67 theme stalk 2190:2194 arg1 domain 2196:2201 the stalk domain 2186:2201 the stalk domain of this protein 2186:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 67 theme stalk 2190:2194 arg1 protein 2211:2217 this protein 2206:2217 this protein 2206:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 11 68 theme glycans 1707:1713 arg1 Sialyation 1666:1675 Sialyation 1666:1675 Sialyation of complex or hybrid N-linked glycans 1666:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 5 69 theme compositions 705:716 arg1 characterisation 674:689 characterisation 674:689 characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species 674:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 13 70 theme glycan 2176:2181 arg1 significance 2147:2158 the functional significance 2132:2158 the functional significance of the O-linked glycan in the stalk domain of this protein 2132:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 10 71 contain containing 1608:1617 arg2 variations 1619:1628 variations 1619:1628 variations of fucose and sulfate or phosphate 1619:1663 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 71 contain containing 1608:1617 arg1 compositions 1595:1606 the compositions 1591:1606 the compositions containing variations of fucose and sulfate or phosphate 1591:1663 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 13 72 theme quantitative 1951:1962 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 73 from domain 2196:2201 arg1 studies 1973:1979 future quantitative glycomic studies 1944:1979 future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 1944:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 73 from domain 2196:2201 arg1 assessment 2118:2127 assessment 2118:2127 assessment of the functional significance of the O-linked glycan in the stalk domain of this protein 2118:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 11 74 located observed 1732:1739 arg2 Sialyation 1666:1675 Sialyation 1666:1675 Sialyation of complex or hybrid N-linked glycans 1666:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 11 74 located observed 1732:1739 arg1 sites 1744:1748 sites 341 and 433 1744:1760 sites 1744:1748 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 11 74 located observed 1732:1739 arg1 433 1758:1760 433 1758:1760 433 1758:1760 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 2 75 theme oligomeric 298:307 arg1 proteins 335:342 oligomeric type II integral membrane proteins 298:342 oligomeric type II integral membrane proteins 298:342 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 75 theme oligomeric 298:307 arg1 glycoproteins 280:292 These glycoproteins 274:292 These glycoproteins 274:292 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 10 76 dep residues 1516:1523 arg1 433 1530:1532 433 1530:1532 433 1530:1532 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 76 dep residues 1516:1523 arg1 341 1525:1527 341 1525:1527 341 1525:1527 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 10 76 dep residues 1516:1523 arg1 481 1538:1540 481 1538:1540 481 1538:1540 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 5 77 theme Avulavirus 766:775 arg1 virus 795:799 the Avulavirus Newcastle disease virus 762:799 the Avulavirus Newcastle disease virus 762:799 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 1 78 theme Rubulavirus 133:143 arg1 genera 145:150 the Avulavirus, Respirovirus and Rubulavirus genera 100:150 the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses 100:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 13 79 theme functional 2136:2145 arg1 significance 2147:2158 the functional significance 2132:2158 the functional significance of the O-linked glycan in the stalk domain of this protein 2132:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 5 80 theme disease 787:793 arg1 virus 795:799 the Avulavirus Newcastle disease virus 762:799 the Avulavirus Newcastle disease virus 762:799 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 9 81 theme high 1431:1434 arg1 glycans 1444:1450 high mannose glycans 1431:1450 high mannose glycans with paucimannose glycans also observed at site 481 1431:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 2 82 theme membrane 326:333 arg1 proteins 335:342 oligomeric type II integral membrane proteins 298:342 oligomeric type II integral membrane proteins 298:342 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 82 theme membrane 326:333 arg1 glycoproteins 280:292 These glycoproteins 274:292 These glycoproteins 274:292 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 12 83 theme protein 1897:1903 arg1 domain 1854:1859 the stalk domain 1844:1859 the stalk domain of the haemagglutinin-neuraminidase protein 1844:1903 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 12 83 theme protein 1897:1903 arg1 protein 1897:1903 the haemagglutinin-neuraminidase protein 1864:1903 the haemagglutinin-neuraminidase protein 1864:1903 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 3 84 attach present 455:461 arg2 glycans 447:453 the N-linked glycans 434:453 the N-linked glycans present on these proteins 434:479 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 3 84 attach present 455:461 arg1 proteins 472:479 these proteins 466:479 these proteins 466:479 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 7 85 gly glycopeptides 1204:1216 arg2 glycopeptides 1204:1216 glycopeptides 1204:1216 glycopeptides from the haemagglutinin-neuraminidase protein 1204:1262 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 7 85 gly glycopeptides 1204:1216 arg1 protein 1256:1262 the haemagglutinin-neuraminidase protein 1223:1262 the haemagglutinin-neuraminidase protein 1223:1262 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 11 86 theme complex 1680:1686 arg1 glycans 1707:1713 complex or hybrid N-linked glycans 1680:1713 glycans 1707:1713 Sialyation of complex or hybrid N-linked glycans was additionally observed at sites 341 and 433. 27928741 1 87 theme Paramyxoviridae 159:173 arg1 family 175:180 the Paramyxoviridae family 155:180 the Paramyxoviridae family of viruses 155:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 5 88 theme species 853:859 arg1 range 838:842 a range 836:842 a range of avian species 836:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 9 89 theme paucimannose 1457:1468 arg1 glycans 1470:1476 paucimannose glycans 1457:1476 paucimannose glycans also observed at site 481 1457:1502 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 3 90 theme Previous 401:408 arg1 studies 410:416 Previous studies 401:416 Previous studies 401:416 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 5 91 attach attached 718:725 arg1 haemagglutinin-neuraminidase 730:757 haemagglutinin-neuraminidase 730:757 haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species 730:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 5 91 attach attached 718:725 arg2 compositions 705:716 the glycan compositions 694:716 the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species 694:859 This study concerns characterisation of the glycan compositions attached to haemagglutinin-neuraminidase of the Avulavirus Newcastle disease virus, which causes Newcastle disease in a range of avian species. 27928741 7 92 theme collision 1113:1121 arg1 dissociation 1123:1134 higher-energy collision dissociation 1099:1134 higher-energy collision dissociation 1099:1134 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 9 93 dep sites 1388:1392 arg1 481 1402:1404 481 1402:1404 481 1402:1404 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 9 93 dep sites 1388:1392 arg1 433 1394:1396 433 1394:1396 433 1394:1396 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 3 94 theme N-linked 438:445 arg1 glycans 447:453 the N-linked glycans 434:453 the N-linked glycans present on these proteins 434:479 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 13 95 theme Newcastle 2061:2069 arg1 virus 2079:2083 Newcastle disease virus 2061:2083 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 96 theme virus 2079:2083 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 3 97 theme virus 513:517 arg1 ability 498:504 the ability 494:504 the ability of the virus to infect host cells and stimulate the host immune system 494:575 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 7 98 theme induced 1077:1083 arg1 dissociation 1085:1096 collision induced dissociation 1067:1096 collision induced dissociation 1067:1096 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 4 99 theme glycans 624:630 arg1 heterogeneity 601:613 site-specific heterogeneity 587:613 site-specific heterogeneity of these glycans 587:630 However, site-specific heterogeneity of these glycans has not been defined. 27928741 10 100 theme sulfate 1644:1650 arg1 variations 1619:1628 variations 1619:1628 variations of fucose and sulfate or phosphate 1619:1663 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 13 101 theme N-linked 2029:2036 arg1 sites 2052:2056 N-linked glycosylation sites 2029:2056 N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 2029:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 101 theme N-linked 2029:2036 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 1 102 theme Respirovirus 116:127 arg1 genera 145:150 the Avulavirus, Respirovirus and Rubulavirus genera 100:150 the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses 100:191 Members of the Avulavirus, Respirovirus and Rubulavirus genera of the Paramyxoviridae family of viruses utilise haemagglutinin-neuraminidase glycoproteins as their attachment proteins. 27928741 12 103 theme undocumented 1789:1800 arg1 glycopeptide 1811:1822 a previously undocumented O-linked glycopeptide 1776:1822 a previously undocumented O-linked glycopeptide 1776:1822 In addition, a previously undocumented O-linked glycopeptide was identified from the stalk domain of the haemagglutinin-neuraminidase protein. 27928741 3 104 theme present 455:461 arg1 glycans 447:453 the N-linked glycans 434:453 the N-linked glycans present on these proteins 434:479 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 0 105 theme disease 45:51 arg1 virus 53:57 the Newcastle disease virus 31:57 the Newcastle disease virus 31:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 2 106 contain possess 351:357 arg2 activity 391:398 haemagglutination and sialidase activity 359:398 haemagglutination and sialidase activity 359:398 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 106 contain possess 351:357 arg1 proteins 335:342 oligomeric type II integral membrane proteins 298:342 oligomeric type II integral membrane proteins 298:342 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 2 106 contain possess 351:357 arg1 glycoproteins 280:292 These glycoproteins 274:292 These glycoproteins 274:292 These glycoproteins are oligomeric type II integral membrane proteins, which possess haemagglutination and sialidase activity. 27928741 6 107 theme avirulent 960:968 arg1 strain 970:975 the avirulent strain QLD/66 956:982 the avirulent strain QLD/66 956:982 Haemagglutinin-neuraminidase was derived from egg propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66. 27928741 0 108 dep haemagglutinin-neuraminidase 59:86 arg1 glycosylation 14:26 Site-specific glycosylation 0:26 Site-specific glycosylation of the Newcastle disease virus 0:57 Site-specific glycosylation of the Newcastle disease virus haemagglutinin-neuraminidase. 27928741 4 109 theme site-specific 587:599 arg1 heterogeneity 601:613 site-specific heterogeneity 587:613 site-specific heterogeneity of these glycans 587:630 However, site-specific heterogeneity of these glycans has not been defined. 27928741 8 110 theme glycan 1288:1293 arg1 compositions 1295:1306 Overall 63, 58, and 37 glycan compositions 1265:1306 Overall 63, 58, and 37 glycan compositions 1265:1306 Overall 63, 58, and 37 glycan compositions were identified at asparagine residues 341, 433 and 481, respectively. 27928741 10 111 theme Asparagine 1505:1514 arg1 residues 1516:1523 Asparagine residues 341, 433 and 481 1505:1540 Asparagine residues 341, 433 and 481 1505:1540 Asparagine residues 341, 433 and 481 contained complex or hybrid glycans with many of the compositions containing variations of fucose and sulfate or phosphate. 27928741 3 112 theme host 558:561 arg1 system 570:575 the host immune system 554:575 the host immune system 554:575 Previous studies have shown that the N-linked glycans present on these proteins can modulate the ability of the virus to infect host cells and stimulate the host immune system. 27928741 7 113 theme mass 1028:1031 arg1 spectrometry 1033:1044 Reverse-phase liquid chromatography tandem mass spectrometry 985:1044 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 13 114 gly glycosylation 2038:2050 arg2 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 114 gly glycosylation 2038:2050 arg1 haemagglutinin-neuraminidase 2085:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 Newcastle disease virus haemagglutinin-neuraminidase 2061:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 114 gly glycosylation 2038:2050 arg2 sites 2052:2056 N-linked glycosylation sites 2029:2056 N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase 2029:2112 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 115 theme protein 2211:2217 arg1 domain 2196:2201 the stalk domain 2186:2201 the stalk domain of this protein 2186:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 13 115 theme protein 2211:2217 arg1 protein 2211:2217 this protein 2206:2217 this protein 2206:2217 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 7 116 theme chromatography 1006:1019 arg1 spectrometry 1033:1044 Reverse-phase liquid chromatography tandem mass spectrometry 985:1044 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 27928741 6 117 theme V4-VAR 934:939 arg1 virions 923:929 propagated virions 912:929 propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66 912:982 Haemagglutinin-neuraminidase was derived from egg propagated virions of V4-VAR, an isolate of the avirulent strain QLD/66. 27928741 9 118 link N-linked 1379:1386 arg1 sites 1388:1392 N-linked sites 1379:1392 N-linked sites 433 and 481 1379:1404 N-linked sites 433 and 481 were observed to contain high mannose glycans with paucimannose glycans also observed at site 481. 27928741 13 119 theme glycan 2004:2009 arg1 structures 2011:2020 glycan structures 2004:2020 glycan structures 2004:2020 These finding will form the basis for future quantitative glycomic studies of the distribution of glycan structures across N-linked glycosylation sites of Newcastle disease virus haemagglutinin-neuraminidase and assessment of the functional significance of the O-linked glycan in the stalk domain of this protein. 27928741 7 120 theme Reverse-phase 985:997 arg1 chromatography 1006:1019 Reverse-phase liquid chromatography 985:1019 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation 985:1169 Reverse-phase liquid chromatography tandem mass spectrometry strategies including collision induced dissociation, higher-energy collision dissociation and electron-transfer dissociation were implemented to characterise glycopeptides from the haemagglutinin-neuraminidase protein. 26567221 0 0 theme HMGB1 64:68 arg1 secretion 51:59 the secretion 47:59 the secretion of HMGB1 47:68 N-linked glycosylation plays a crucial role in the secretion of HMGB1. 26567221 6 1 gly Non-glycosylated 917:932 arg1 N→Q 949:951 N→Q 949:951 N→Q 949:951 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 1 gly Non-glycosylated 917:932 arg1 mutant 941:946 Non-glycosylated double mutant 917:946 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 5 2 theme HMGB1 875:879 arg1 shift 866:870 a molecular shift 854:870 a molecular shift of HMGB1 854:879 Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. 26567221 4 3 theme chromatography 690:703 arg1 LC-MS/MS 731:738 LC-MS/MS 731:738 LC-MS/MS 731:738 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 3 theme chromatography 690:703 arg1 spectrometry 717:728 liquid chromatography tandem mass spectrometry 683:728 liquid chromatography tandem mass spectrometry (LC-MS/MS) 683:739 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 1 4 dep stimulants 185:194 arg1 response 165:172 response 165:172 response 165:172 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 3 5 theme N-glycosylation 506:520 arg1 role 498:501 the role 494:501 the role of N-glycosylation of HMGB1 in extracellular secretion 494:556 Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. 26567221 4 6 theme tandem 705:710 arg1 LC-MS/MS 731:738 LC-MS/MS 731:738 LC-MS/MS 731:738 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 6 theme tandem 705:710 arg1 spectrometry 717:728 liquid chromatography tandem mass spectrometry 683:728 liquid chromatography tandem mass spectrometry (LC-MS/MS) 683:739 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 3 7 theme extracellular 534:546 arg1 secretion 548:556 extracellular secretion 534:556 extracellular secretion 534:556 Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. 26567221 8 8 theme extracellular 1465:1477 arg1 secretion 1479:1487 extracellular secretion 1465:1487 extracellular secretion 1465:1487 Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion. 26567221 6 9 theme protein 1104:1110 arg1 CRM1 1112:1115 the nuclear export protein CRM1 1085:1115 the nuclear export protein CRM1 1085:1115 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 2 10 theme -targeting 283:292 arg1 peptide 301:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 7 11 theme reduced 1184:1190 arg1 secretion 1192:1200 reduced secretion 1184:1200 reduced secretion 1184:1200 These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. 26567221 0 12 link N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation plays a crucial role in the secretion of HMGB1. 26567221 6 13 theme export 1097:1102 arg1 CRM1 1112:1115 the nuclear export protein CRM1 1085:1115 the nuclear export protein CRM1 1085:1115 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 7 14 theme pro-inflammatory 1269:1284 arg1 stimuli 1286:1292 pro-inflammatory stimuli 1269:1292 pro-inflammatory stimuli 1269:1292 These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. 26567221 6 15 theme HMGB1 954:958 arg1 HMGB1 992:996 HMGB1 992:996 HMGB1(N37Q/N135Q) 992:1008 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 15 theme HMGB1 954:958 arg1 HMGB1 970:974 HMGB1 970:974 HMGB1(N37Q/N134Q) 970:986 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 15 theme HMGB1 954:958 arg1 proteins 960:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins 917:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 16 theme nuclear 1089:1095 arg1 CRM1 1112:1115 the nuclear export protein CRM1 1085:1115 the nuclear export protein CRM1 1085:1115 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 1 17 theme extracellular 141:153 arg1 milieu 155:160 the extracellular milieu 137:160 the extracellular milieu 137:160 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 2 18 theme post-translational 440:457 arg1 modifications 459:471 post-translational modifications 440:471 post-translational modifications 440:471 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 4 19 theme non-consensus 605:617 arg1 residues 626:633 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 residues 626:633 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 1 20 theme HMGB1 71:75 arg1 protein 77:83 HMGB1 protein 71:83 HMGB1 protein 71:83 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 1 20 theme HMGB1 71:75 arg1 mediator 98:105 a delayed mediator 88:105 a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response 88:232 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 2 21 theme extracellular 334:346 arg1 secretion 348:356 extracellular secretion 334:356 extracellular secretion 334:356 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 0 22 theme N-linked 0:7 arg1 glycosylation 9:21 N-linked glycosylation 0:21 N-linked glycosylation 0:21 N-linked glycosylation plays a crucial role in the secretion of HMGB1. 26567221 4 23 theme liquid 683:688 arg1 LC-MS/MS 731:738 LC-MS/MS 731:738 LC-MS/MS 731:738 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 23 theme liquid 683:688 arg1 spectrometry 717:728 liquid chromatography tandem mass spectrometry 683:728 liquid chromatography tandem mass spectrometry (LC-MS/MS) 683:739 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 24 theme N135 620:623 arg1 residues 626:633 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 residues 626:633 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 5 25 theme N-glycosylation 809:823 arg1 Inhibition 795:804 Inhibition 795:804 Inhibition of N-glycosylation with tunicamycin 795:840 Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. 26567221 6 26 theme rapid 1121:1125 arg1 degradation 1127:1137 rapid degradation 1121:1137 rapid degradation by ubiquitylation 1121:1155 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 5 27 theme gel 896:898 arg1 electrophoresis 900:914 gel electrophoresis 896:914 gel electrophoresis 896:914 Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. 26567221 3 28 theme HMGB1 525:529 arg1 N-glycosylation 506:520 N-glycosylation 506:520 N-glycosylation of HMGB1 506:529 Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. 26567221 2 29 theme reticulum 269:277 arg1 peptide 301:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 6 30 theme strong 1046:1051 arg1 binding 1053:1059 strong binding 1046:1059 strong binding to DNA 1046:1066 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 7 31 contain had 1180:1182 arg1 proteins 1171:1178 These mutant proteins 1158:1178 These mutant proteins 1158:1178 These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. 26567221 7 31 contain had 1180:1182 arg2 secretion 1192:1200 reduced secretion 1184:1200 reduced secretion 1184:1200 These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. 26567221 6 32 theme mutant 941:946 arg1 HMGB1 992:996 HMGB1 992:996 HMGB1(N37Q/N135Q) 992:1008 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 32 theme mutant 941:946 arg1 HMGB1 970:974 HMGB1 970:974 HMGB1(N37Q/N134Q) 970:986 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 32 theme mutant 941:946 arg1 proteins 960:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins 917:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 5 33 with Inhibition 795:804 arg1 tunicamycin 830:840 tunicamycin 830:840 tunicamycin 830:840 Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. 26567221 4 34 theme one 601:603 arg1 residues 626:633 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 residues 626:633 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 1 35 theme various 177:183 arg1 stimulants 185:194 various stimulants 177:194 various stimulants 177:194 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 0 36 theme crucial 31:37 arg1 role 39:42 a crucial role 29:42 a crucial role 29:42 N-linked glycosylation plays a crucial role in the secretion of HMGB1. 26567221 6 37 theme double 934:939 arg1 N→Q 949:951 N→Q 949:951 N→Q 949:951 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 37 theme double 934:939 arg1 mutant 941:946 Non-glycosylated double mutant 917:946 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 38 theme Non-glycosylated 917:932 arg1 N→Q 949:951 N→Q 949:951 N→Q 949:951 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 38 theme Non-glycosylated 917:932 arg1 mutant 941:946 Non-glycosylated double mutant 917:946 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 8 39 theme DNA 1387:1389 arg1 interaction 1391:1401 its DNA interaction 1383:1401 its DNA interaction 1383:1401 Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion. 26567221 1 40 theme delayed 90:96 arg1 protein 77:83 HMGB1 protein 71:83 HMGB1 protein 71:83 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 1 40 theme delayed 90:96 arg1 mediator 98:105 a delayed mediator 88:105 a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response 88:232 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 8 41 theme nucleocytoplasmic 1433:1449 arg1 transport 1451:1459 its nucleocytoplasmic transport 1429:1459 its nucleocytoplasmic transport 1429:1459 Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion. 26567221 6 42 dep proteins 960:967 arg1 N37Q/N135Q 998:1007 N37Q/N135Q 998:1007 N37Q/N135Q 998:1007 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 42 dep proteins 960:967 arg1 N37Q/N134Q 976:985 N37Q/N134Q 976:985 N37Q/N134Q 976:985 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 42 dep proteins 960:967 arg1 HMGB1 992:996 HMGB1 992:996 HMGB1(N37Q/N135Q) 992:1008 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 42 dep proteins 960:967 arg1 HMGB1 970:974 HMGB1 970:974 HMGB1(N37Q/N134Q) 970:986 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 6 42 dep proteins 960:967 arg1 proteins 960:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins 917:967 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) 917:1009 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 2 43 theme peptide 301:307 arg1 devoid 244:249 devoid 244:249 devoid 244:249 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 2 44 theme signal 294:299 arg1 peptide 301:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 4 45 theme mass 712:715 arg1 LC-MS/MS 731:738 LC-MS/MS 731:738 LC-MS/MS 731:738 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 45 theme mass 712:715 arg1 spectrometry 717:728 liquid chromatography tandem mass spectrometry 683:728 liquid chromatography tandem mass spectrometry (LC-MS/MS) 683:739 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 46 gly N-glycosylated 645:658 arg1 HMGB1 663:667 HMGB1 663:667 HMGB1 663:667 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 46 gly N-glycosylated 645:658 arg1 residues 626:633 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 residues 626:633 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 46 gly N-glycosylated 645:658 arg2 consensus 572:580 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 consensus 572:580 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 46 gly N-glycosylated 645:658 arg1 consensus 572:580 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 consensus 572:580 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 46 gly N-glycosylated 645:658 arg2 residues 626:633 two consensus (N37 and N134) and one non-consensus (N135) residues 568:633 residues 626:633 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 2 47 theme endoplasmic 257:267 arg1 ER 280:281 ER 280:281 ER 280:281 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 2 47 theme endoplasmic 257:267 arg1 reticulum 269:277 an endoplasmic reticulum 254:277 an endoplasmic reticulum (ER)-targeting signal peptide 254:307 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 1 48 theme sepsis 110:115 arg1 protein 77:83 HMGB1 protein 71:83 HMGB1 protein 71:83 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 1 48 theme sepsis 110:115 arg1 mediator 98:105 a delayed mediator 88:105 a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response 88:232 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 26567221 4 49 from HMGB1 663:667 arg1 N-glycosylated 645:658 N-glycosylated 645:658 N-glycosylated 645:658 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 8 50 gly N-glycosylated 1336:1349 arg1 HMGB1 1327:1331 HMGB1 1327:1331 HMGB1 1327:1331 Taken together, we propose that HMGB1 is N-glycosylated, and that this is important for its DNA interaction and is a prerequisite for its nucleocytoplasmic transport and extracellular secretion. 26567221 4 51 dep consensus 572:580 arg1 N134 591:594 N134 591:594 N134 591:594 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 51 dep consensus 572:580 arg1 N37 583:585 N37 583:585 N37 583:585 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 52 theme N-glycan 759:766 arg1 composition 768:778 N-glycan composition 759:778 N-glycan composition 759:778 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 4 53 from N-glycosylated 645:658 arg1 HMGB1 663:667 HMGB1 663:667 HMGB1 663:667 We found two consensus (N37 and N134) and one non-consensus (N135) residues that were N-glycosylated in HMGB1 by performing liquid chromatography tandem mass spectrometry (LC-MS/MS) and analyzing for N-glycan composition and structure. 26567221 5 54 theme molecular 856:864 arg1 shift 866:870 a molecular shift 854:870 a molecular shift of HMGB1 854:879 Inhibition of N-glycosylation with tunicamycin resulted in a molecular shift of HMGB1 as assessed by gel electrophoresis. 26567221 2 55 theme secretion 348:356 arg1 mechanism 321:329 the mechanism 317:329 the mechanism of extracellular secretion 317:356 HMGB1 is devoid of an endoplasmic reticulum (ER)-targeting signal peptide; hence, the mechanism of extracellular secretion is not completely understood, although HMGB1 is secreted after being subjected to post-translational modifications. 26567221 7 56 theme mutant 1164:1169 arg1 proteins 1171:1178 These mutant proteins 1158:1178 These mutant proteins 1158:1178 These mutant proteins had reduced secretion even after acetylation, phosphorylation, oxidation and exposure to pro-inflammatory stimuli. 26567221 6 57 theme weak 1069:1072 arg1 binding 1074:1080 weak binding 1069:1080 weak binding to the nuclear export protein CRM1 1069:1115 Non-glycosylated double mutant (N→Q) HMGB1 proteins (HMGB1(N37Q/N134Q) and HMGB1(N37Q/N135Q)) showed localization to the nuclei, strong binding to DNA, weak binding to the nuclear export protein CRM1 and rapid degradation by ubiquitylation. 26567221 3 58 gly N-glycosylation 506:520 arg1 HMGB1 525:529 HMGB1 525:529 HMGB1 525:529 Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. 26567221 3 59 from role 498:501 arg1 secretion 548:556 extracellular secretion 534:556 extracellular secretion 534:556 Here, we identified the role of N-glycosylation of HMGB1 in extracellular secretion. 26567221 1 60 theme pro-inflammatory 208:223 arg1 response 225:232 a pro-inflammatory response 206:232 a pro-inflammatory response 206:232 HMGB1 protein is a delayed mediator of sepsis that is secreted to the extracellular milieu in response to various stimulants, inducing a pro-inflammatory response. 27732771 4 0 theme N-linked 409:416 arg1 trimming 425:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 16 1 theme VWF 2330:2332 arg1 glycosylation 2334:2346 VWF glycosylation 2330:2346 VWF glycosylation 2330:2346 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 8 2 theme molecular 973:981 arg1 mechanisms 983:992 the molecular mechanisms 969:992 the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance 969:1065 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 2 3 theme glycan 255:260 arg1 moieties 262:269 specific glycan moieties 246:269 specific glycan moieties 246:269 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 5 4 theme Willebrand 535:544 arg1 factor 546:551 von Willebrand factor 531:551 von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 531:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 5 from presentation 563:574 arg1 clearance 518:526 Dr Denis discuss clearance 501:526 Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 501:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 6 6 theme previous 763:770 arg1 studies 772:778 previous studies 763:778 previous studies 763:778 In addition, previous studies have demonstrated that VWF glycans play a key role in regulating in vivo clearance. 27732771 5 7 theme Dr 501:502 arg1 clearance 518:526 Dr Denis discuss clearance 501:526 Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 501:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 10 8 gly asialoglycoprotein 1295:1312 arg1 asialoglycoprotein 1295:1312 asialoglycoprotein receptor 1295:1321 asialoglycoprotein receptor (ASGPR) 1295:1329 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 8 9 theme VWF 1008:1010 arg1 structures 1028:1037 VWF N-linked glycan structures 1008:1037 VWF N-linked glycan structures 1008:1037 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 2 10 used used 218:221 arg2 glycoforms 202:211 VWF glycoforms 198:211 VWF glycoforms 198:211 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 13 11 link N-linked 1698:1705 arg1 effects 1714:1720 these additional N-linked glycan effects 1681:1720 these additional N-linked glycan effects on clearance 1681:1733 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 5 12 theme type 713:716 arg1 etiology 685:692 the etiology 681:692 the etiology of both type 1 and type 2 von Willebrand disease (VWD) 681:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 10 13 theme receptor 1314:1321 arg1 inhibitors 1281:1290 inhibitors 1281:1290 inhibitors of asialoglycoprotein receptor (ASGPR) 1281:1329 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 11 14 theme pd-VWF 1468:1473 arg1 clearance 1475:1483 enhanced pd-VWF clearance 1459:1483 enhanced pd-VWF clearance modulated via ASGPR 1459:1503 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 11 15 dep Results 1386:1392 arg1 resulted 1447:1454 resulted 1447:1454 resulted in enhanced pd-VWF clearance modulated via ASGPR 1447:1503 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 5 16 theme von 720:722 arg1 VWD 744:746 VWD 744:746 VWD 744:746 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 16 theme von 720:722 arg1 disease 735:741 von Willebrand disease 720:741 von Willebrand disease (VWD) 720:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 15 17 theme ASGPR-dependent 2206:2220 arg1 pathways 2247:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 11 18 theme N-linked 1413:1420 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 13 19 theme enhanced 1780:1787 arg1 clearance 1800:1808 enhanced macrophage clearance 1780:1808 enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1 1780:1884 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 5 20 theme ISTH 585:588 arg1 SUMMARY 598:604 the ISTH Academy SUMMARY 581:604 the ISTH Academy SUMMARY 581:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 12 21 theme glycan 1602:1607 arg1 trimming 1609:1616 progressive N-linked glycan trimming 1581:1616 progressive N-linked glycan trimming 1581:1616 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 2 22 gly glycoforms 202:211 arg1 VWF 198:200 VWF glycoforms 198:211 VWF glycoforms 198:211 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 13 23 theme receptor-related 1859:1874 arg1 protein 1876:1882 LDL receptor-related protein 1 1855:1884 LDL receptor-related protein 1 1855:1884 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 4 24 theme macrophage-mediated 456:474 arg1 clearance 476:484 increased macrophage-mediated clearance 446:484 increased macrophage-mediated clearance 446:484 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 5 25 dep clearance 518:526 arg1 discuss 510:516 discuss 510:516 discuss 510:516 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 0 26 theme von 71:73 arg1 factor 86:91 plasma-derived von Willebrand factor 56:91 plasma-derived von Willebrand factor 56:91 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 5 27 theme free 558:561 arg1 presentation 563:574 a free presentation 556:574 a free presentation from the ISTH Academy SUMMARY 556:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 16 28 from important 2355:2363 arg1 pathophysiology 2372:2386 the pathophysiology 2368:2386 the pathophysiology underlying type 1C VWD 2368:2409 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 16 29 theme 1C 2404:2405 arg1 VWD 2407:2409 type 1C VWD 2399:2409 type 1C VWD 2399:2409 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 0 30 theme factor 86:91 arg1 clearance 43:51 clearance 43:51 clearance of plasma-derived von Willebrand factor 43:91 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 8 31 theme glycan 1021:1026 arg1 structures 1028:1037 VWF N-linked glycan structures 1008:1037 VWF N-linked glycan structures 1008:1037 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 5 32 theme von 627:629 arg1 VWF 650:652 VWF 650:652 VWF 650:652 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 32 theme von 627:629 arg1 factor 642:647 von Willebrand factor 627:647 Enhanced von Willebrand factor (VWF) clearance 618:663 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 33 dep type 702:705 arg1 VWD 744:746 VWD 744:746 VWD 744:746 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 33 dep type 702:705 arg1 disease 735:741 von Willebrand disease 720:741 von Willebrand disease (VWD) 720:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 34 dep hear 496:499 arg1 Background 607:616 Background 607:616 Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD) 607:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 10 35 theme VWF-/- 1239:1244 arg1 mice 1246:1249 VWF-/- mice 1239:1249 VWF-/- mice 1239:1249 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 11 36 link O-linked 1426:1433 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 5 37 theme factor 642:647 arg1 clearance 655:663 Enhanced von Willebrand factor (VWF) clearance 618:663 Enhanced von Willebrand factor (VWF) clearance 618:663 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 11 38 theme enhanced 1459:1466 arg1 clearance 1475:1483 enhanced pd-VWF clearance 1459:1483 enhanced pd-VWF clearance modulated via ASGPR 1459:1503 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 12 39 theme sialylation 1543:1553 arg1 role 1526:1529 this role 1521:1529 this role of terminal sialylation 1521:1553 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 3 40 theme VWF 342:344 arg1 clearance 346:354 enhanced VWF clearance 333:354 enhanced VWF clearance through asialoglycoprotein receptor 333:390 Reduction in sialylation resulted in enhanced VWF clearance through asialoglycoprotein receptor. 27732771 13 41 theme N-linked 1698:1705 arg1 effects 1714:1720 these additional N-linked glycan effects 1681:1720 these additional N-linked glycan effects on clearance 1681:1733 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 11 42 theme sialylation 1435:1445 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 11 42 theme sialylation 1435:1445 arg1 amounts 1402:1408 Reduced amounts 1394:1408 Reduced amounts of N-linked and O-linked sialylation 1394:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 0 43 theme N-linked 0:7 arg1 causes 27:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 13 44 from effects 1714:1720 arg1 clearance 1725:1733 clearance 1725:1733 clearance 1725:1733 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 7 45 theme VWF 909:911 arg1 clearance 913:921 VWF clearance 909:921 VWF clearance 909:921 However, the molecular mechanisms underlying VWF clearance remain poorly understood. 27732771 1 46 theme von 105:107 arg1 VWF 129:131 VWF 129:131 VWF 129:131 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 1 46 theme von 105:107 arg1 factor 121:126 Essentials von Willebrands factor 94:126 Essentials von Willebrands factor (VWF) glycosylation 94:146 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 12 47 theme VWF 1653:1655 arg1 clearance 1657:1665 markedly enhanced VWF clearance 1635:1665 markedly enhanced VWF clearance 1635:1665 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 15 48 theme carbohydrate 2069:2080 arg1 determinants 2082:2093 non-sialic acid carbohydrate determinants 2053:2093 non-sialic acid carbohydrate determinants expressed on VWF 2053:2110 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 0 49 link N-linked 0:7 arg1 causes 27:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 10 50 theme asialoglycoprotein 1295:1312 arg1 ASGPR 1324:1328 ASGPR 1324:1328 ASGPR 1324:1328 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 10 50 theme asialoglycoprotein 1295:1312 arg1 receptor 1314:1321 asialoglycoprotein receptor 1295:1321 asialoglycoprotein receptor (ASGPR) 1295:1329 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 0 51 theme truncation 16:25 arg1 causes 27:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 1 52 theme factor 121:126 arg1 glycosylation 134:146 Essentials von Willebrands factor (VWF) glycosylation 94:146 Essentials von Willebrands factor (VWF) glycosylation 94:146 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 15 53 theme non-sialic 2053:2062 arg1 determinants 2082:2093 non-sialic acid carbohydrate determinants 2053:2093 non-sialic acid carbohydrate determinants expressed on VWF 2053:2110 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 1 54 dep in 179:180 arg1 vivo 182:185 vivo 182:185 vivo 182:185 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 10 55 theme inhibitors 1281:1290 arg1 absence 1270:1276 absence 1270:1276 absence 1270:1276 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 10 55 theme inhibitors 1281:1290 arg1 presence 1258:1265 presence 1258:1265 presence 1258:1265 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 5 56 from clearance 518:526 arg1 presentation 563:574 a free presentation 556:574 a free presentation from the ISTH Academy SUMMARY 556:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 15 57 dep in 2167:2168 arg1 vivo 2170:2173 vivo 2170:2173 vivo 2170:2173 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 9 58 theme plasma-derived 1125:1138 arg1 exoglycosidases 1098:1112 exoglycosidases 1098:1112 exoglycosidases 1098:1112 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 58 theme plasma-derived 1125:1138 arg1 glycoforms 1153:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 15 59 theme acid 2064:2067 arg1 determinants 2082:2093 non-sialic acid carbohydrate determinants 2053:2093 non-sialic acid carbohydrate determinants expressed on VWF 2053:2110 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 5 60 from important 668:676 arg1 etiology 685:692 the etiology 681:692 the etiology of both type 1 and type 2 von Willebrand disease (VWD) 681:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 16 61 from variation 2317:2325 arg1 glycosylation 2334:2346 VWF glycosylation 2330:2346 VWF glycosylation 2330:2346 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 9 62 link plasma-derived 1125:1138 arg1 exoglycosidases 1098:1112 exoglycosidases 1098:1112 exoglycosidases 1098:1112 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 62 link plasma-derived 1125:1138 arg1 glycoforms 1153:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 2 63 theme VWF 198:200 arg1 glycoforms 202:211 VWF glycoforms 198:211 VWF glycoforms 198:211 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 5 64 from factor 546:551 arg1 SUMMARY 598:604 the ISTH Academy SUMMARY 581:604 the ISTH Academy SUMMARY 581:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 64 from factor 546:551 arg1 presentation 563:574 a free presentation 556:574 a free presentation from the ISTH Academy SUMMARY 556:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 12 65 link N-linked 1593:1600 arg1 trimming 1609:1616 progressive N-linked glycan trimming 1581:1616 progressive N-linked glycan trimming 1581:1616 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 10 66 theme clodronate-induced 1345:1362 arg1 depletion 1375:1383 clodronate-induced macrophage depletion 1345:1383 clodronate-induced macrophage depletion 1345:1383 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 14 67 dep Conclusion 1887:1896 arg1 regulate 1945:1952 regulate 1945:1952 regulate susceptibility to proteolysis by ADAMTS-13 1945:1995 Conclusion The carbohydrate determinants expressed on VWF regulate susceptibility to proteolysis by ADAMTS-13. 27732771 7 68 theme molecular 877:885 arg1 mechanisms 887:896 the molecular mechanisms 873:896 the molecular mechanisms underlying VWF clearance 873:921 However, the molecular mechanisms underlying VWF clearance remain poorly understood. 27732771 3 69 from Reduction 296:304 arg1 sialylation 309:319 sialylation 309:319 sialylation 309:319 Reduction in sialylation resulted in enhanced VWF clearance through asialoglycoprotein receptor. 27732771 5 70 dep Background 607:616 arg1 important 668:676 important 668:676 important 668:676 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 10 71 theme glycoforms 1207:1216 arg1 clearance 1188:1196 In vivo clearance 1180:1196 In vivo clearance of these glycoforms 1180:1216 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 4 72 theme glycan 418:423 arg1 trimming 425:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 8 73 dep in 1049:1050 arg1 vivo 1052:1055 vivo 1052:1055 vivo 1052:1055 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 6 74 theme VWF 803:805 arg1 glycans 807:813 VWF glycans 803:813 VWF glycans 803:813 In addition, previous studies have demonstrated that VWF glycans play a key role in regulating in vivo clearance. 27732771 2 75 theme moieties 262:269 arg1 role 238:241 the role 234:241 the role of specific glycan moieties in regulating clearance 234:293 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 5 76 theme von 531:533 arg1 factor 546:551 von Willebrand factor 531:551 von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 531:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 9 77 gly glycoforms 1153:1162 arg1 VWF 1140:1142 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 77 gly glycoforms 1153:1162 arg1 pd-VWF 1145:1150 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 0 78 link plasma-derived 56:69 arg1 factor 86:91 plasma-derived von Willebrand factor 56:91 plasma-derived von Willebrand factor 56:91 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 13 79 theme macrophage 1789:1798 arg1 clearance 1800:1808 enhanced macrophage clearance 1780:1808 enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1 1780:1884 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 5 80 theme factor 546:551 arg1 clearance 518:526 Dr Denis discuss clearance 501:526 Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 501:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 1 81 theme in 179:180 arg1 clearance 187:195 in vivo clearance 179:195 in vivo clearance 179:195 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 4 82 theme VWF 405:407 arg1 trimming 425:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 8 83 theme N-linked 1012:1019 arg1 structures 1028:1037 VWF N-linked glycan structures 1008:1037 VWF N-linked glycan structures 1008:1037 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 5 84 theme Denis 504:508 arg1 clearance 518:526 Dr Denis discuss clearance 501:526 Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 501:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 15 85 theme hepatic 2198:2204 arg1 pathways 2247:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 12 86 theme N-linked 1593:1600 arg1 trimming 1609:1616 progressive N-linked glycan trimming 1581:1616 progressive N-linked glycan trimming 1581:1616 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 6 87 dep in 845:846 arg1 vivo 848:851 vivo 848:851 vivo 848:851 In addition, previous studies have demonstrated that VWF glycans play a key role in regulating in vivo clearance. 27732771 14 88 theme carbohydrate 1902:1913 arg1 determinants 1915:1926 The carbohydrate determinants 1898:1926 The carbohydrate determinants expressed on VWF 1898:1943 Conclusion The carbohydrate determinants expressed on VWF regulate susceptibility to proteolysis by ADAMTS-13. 27732771 4 89 theme increased 446:454 arg1 clearance 476:484 increased macrophage-mediated clearance 446:484 increased macrophage-mediated clearance 446:484 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 4 90 theme Progressive 393:403 arg1 trimming 425:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 5 91 theme type 702:705 arg1 etiology 685:692 the etiology 681:692 the etiology of both type 1 and type 2 von Willebrand disease (VWD) 681:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 2 92 theme specific 246:253 arg1 moieties 262:269 specific glycan moieties 246:269 specific glycan moieties 246:269 VWF glycoforms were used to examine the role of specific glycan moieties in regulating clearance. 27732771 5 93 theme Academy 590:596 arg1 SUMMARY 598:604 the ISTH Academy SUMMARY 581:604 the ISTH Academy SUMMARY 581:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 94 from SUMMARY 598:604 arg1 presentation 563:574 a free presentation 556:574 a free presentation from the ISTH Academy SUMMARY 556:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 94 from SUMMARY 598:604 arg1 factor 546:551 von Willebrand factor 531:551 von Willebrand factor in a free presentation from the ISTH Academy SUMMARY 531:604 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 16 95 theme type 2399:2402 arg1 VWD 2407:2409 type 1C VWD 2399:2409 type 1C VWD 2399:2409 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 0 96 theme Willebrand 75:84 arg1 factor 86:91 plasma-derived von Willebrand factor 56:91 plasma-derived von Willebrand factor 56:91 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 10 97 dep presence 1258:1265 arg1 the 1254:1256 the 1254:1256 the 1254:1256 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 3 98 theme enhanced 333:340 arg1 clearance 346:354 enhanced VWF clearance 333:354 enhanced VWF clearance through asialoglycoprotein receptor 333:390 Reduction in sialylation resulted in enhanced VWF clearance through asialoglycoprotein receptor. 27732771 1 99 theme key 156:158 arg1 role 160:163 a key role 154:163 a key role 154:163 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 12 100 theme terminal 1534:1541 arg1 sialylation 1543:1553 terminal sialylation 1534:1553 terminal sialylation 1534:1553 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 5 101 theme Willebrand 724:733 arg1 VWD 744:746 VWD 744:746 VWD 744:746 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 101 theme Willebrand 724:733 arg1 disease 735:741 von Willebrand disease 720:741 von Willebrand disease (VWD) 720:747 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 15 102 theme in 2167:2168 arg1 clearance 2175:2183 in vivo clearance 2167:2183 in vivo clearance 2167:2183 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 8 103 theme in 1049:1050 arg1 clearance 1057:1065 in vivo clearance 1049:1065 in vivo clearance 1049:1065 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 5 104 theme Willebrand 631:640 arg1 VWF 650:652 VWF 650:652 VWF 650:652 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 5 104 theme Willebrand 631:640 arg1 factor 642:647 von Willebrand factor 627:647 Enhanced von Willebrand factor (VWF) clearance 618:663 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 3 105 theme asialoglycoprotein 364:381 arg1 receptor 383:390 asialoglycoprotein receptor 364:390 asialoglycoprotein receptor 364:390 Reduction in sialylation resulted in enhanced VWF clearance through asialoglycoprotein receptor. 27732771 15 106 theme important 2138:2146 arg1 role 2148:2151 an unexpectedly important role 2122:2151 an unexpectedly important role 2122:2151 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 15 107 theme macrophage-dependent 2226:2245 arg1 pathways 2247:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 both hepatic ASGPR-dependent and macrophage-dependent pathways 2193:2254 In addition, our findings now further demonstrate that non-sialic acid carbohydrate determinants expressed on VWF also play an unexpectedly important role in modulating in vivo clearance through both hepatic ASGPR-dependent and macrophage-dependent pathways. 27732771 11 108 link N-linked 1413:1420 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 13 109 theme glycan 1707:1712 arg1 effects 1714:1720 these additional N-linked glycan effects 1681:1720 these additional N-linked glycan effects on clearance 1681:1733 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 11 110 theme O-linked 1426:1433 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 0 111 theme glycan 9:14 arg1 causes 27:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes 0:32 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 1 112 theme Essentials 94:103 arg1 VWF 129:131 VWF 129:131 VWF 129:131 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 1 112 theme Essentials 94:103 arg1 factor 121:126 Essentials von Willebrands factor 94:126 Essentials von Willebrands factor (VWF) glycosylation 94:146 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 9 113 theme series 1088:1093 arg1 use 1079:1081 use 1079:1081 use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms 1079:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 114 theme pd-VWF 1145:1150 arg1 exoglycosidases 1098:1112 exoglycosidases 1098:1112 exoglycosidases 1098:1112 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 114 theme pd-VWF 1145:1150 arg1 glycoforms 1153:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 3 115 gly asialoglycoprotein 364:381 arg1 asialoglycoprotein 364:381 asialoglycoprotein receptor 364:390 asialoglycoprotein receptor 364:390 Reduction in sialylation resulted in enhanced VWF clearance through asialoglycoprotein receptor. 27732771 5 116 theme Enhanced 618:625 arg1 clearance 655:663 Enhanced von Willebrand factor (VWF) clearance 618:663 Enhanced von Willebrand factor (VWF) clearance 618:663 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 1 117 theme Willebrands 109:119 arg1 VWF 129:131 VWF 129:131 VWF 129:131 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 1 117 theme Willebrands 109:119 arg1 factor 121:126 Essentials von Willebrands factor 94:126 Essentials von Willebrands factor (VWF) glycosylation 94:146 Essentials von Willebrands factor (VWF) glycosylation plays a key role in modulating in vivo clearance. 27732771 12 118 theme enhanced 1644:1651 arg1 clearance 1657:1665 markedly enhanced VWF clearance 1635:1665 markedly enhanced VWF clearance 1635:1665 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 27732771 16 119 from pathophysiology 2372:2386 arg1 important 2355:2363 important 2355:2363 important 2355:2363 In addition, these data further support the hypothesis that variation in VWF glycosylation may be important in the pathophysiology underlying type 1C VWD. 27732771 9 120 theme exoglycosidases 1098:1112 arg1 series 1088:1093 a series 1086:1093 a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms 1086:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 8 121 link N-linked 1012:1019 arg1 structures 1028:1037 VWF N-linked glycan structures 1008:1037 VWF N-linked glycan structures 1008:1037 Objective To define the molecular mechanisms through which VWF N-linked glycan structures influence in vivo clearance. 27732771 6 122 theme in 845:846 arg1 clearance 853:861 in vivo clearance 845:861 in vivo clearance 845:861 In addition, previous studies have demonstrated that VWF glycans play a key role in regulating in vivo clearance. 27732771 4 123 link N-linked 409:416 arg1 trimming 425:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming 393:432 Progressive VWF N-linked glycan trimming resulted in increased macrophage-mediated clearance. 27732771 13 124 theme additional 1687:1696 arg1 effects 1714:1720 these additional N-linked glycan effects 1681:1720 these additional N-linked glycan effects on clearance 1681:1733 Furthermore, these additional N-linked glycan effects on clearance were ASGPR-independent, and instead involved enhanced macrophage clearance that was mediated, at least in part, through LDL receptor-related protein 1. 27732771 9 125 theme different 1115:1123 arg1 exoglycosidases 1098:1112 exoglycosidases 1098:1112 exoglycosidases 1098:1112 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 125 theme different 1115:1123 arg1 glycoforms 1153:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 0 126 theme plasma-derived 56:69 arg1 factor 86:91 plasma-derived von Willebrand factor 56:91 plasma-derived von Willebrand factor 56:91 N-linked glycan truncation causes enhanced clearance of plasma-derived von Willebrand factor. 27732771 5 127 from etiology 685:692 arg1 important 668:676 important 668:676 important 668:676 Click to hear Dr Denis discuss clearance of von Willebrand factor in a free presentation from the ISTH Academy SUMMARY: Background Enhanced von Willebrand factor (VWF) clearance is important in the etiology of both type 1 and type 2 von Willebrand disease (VWD). 27732771 9 128 theme VWF 1140:1142 arg1 exoglycosidases 1098:1112 exoglycosidases 1098:1112 exoglycosidases 1098:1112 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 9 128 theme VWF 1140:1142 arg1 glycoforms 1153:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 different plasma-derived VWF (pd-VWF) glycoforms 1115:1162 Methods By use of a series of exoglycosidases, different plasma-derived VWF (pd-VWF) glycoforms were generated. 27732771 6 129 theme key 822:824 arg1 role 826:829 a key role 820:829 a key role 820:829 In addition, previous studies have demonstrated that VWF glycans play a key role in regulating in vivo clearance. 27732771 10 130 theme macrophage 1364:1373 arg1 depletion 1375:1383 clodronate-induced macrophage depletion 1345:1383 clodronate-induced macrophage depletion 1345:1383 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 10 131 theme In 1180:1181 arg1 clearance 1188:1196 In vivo clearance 1180:1196 In vivo clearance of these glycoforms 1180:1216 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 10 132 dep In 1180:1181 arg1 vivo 1183:1186 vivo 1183:1186 vivo 1183:1186 In vivo clearance of these glycoforms was then assessed in VWF-/- mice in the presence or absence of inhibitors of asialoglycoprotein receptor (ASGPR), or following clodronate-induced macrophage depletion. 27732771 11 133 theme Reduced 1394:1400 arg1 sialylation 1435:1445 N-linked and O-linked sialylation 1413:1445 N-linked and O-linked sialylation 1413:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 11 133 theme Reduced 1394:1400 arg1 amounts 1402:1408 Reduced amounts 1394:1408 Reduced amounts of N-linked and O-linked sialylation 1394:1445 Results Reduced amounts of N-linked and O-linked sialylation resulted in enhanced pd-VWF clearance modulated via ASGPR. 27732771 12 134 theme progressive 1581:1591 arg1 trimming 1609:1616 progressive N-linked glycan trimming 1581:1616 progressive N-linked glycan trimming 1581:1616 In addition to this role of terminal sialylation, we further observed that progressive N-linked glycan trimming also resulted in markedly enhanced VWF clearance. 28331984 6 0 theme residue 708:714 arg1 3OMe-L-rhamnose 716:730 the residue 3OMe-L-rhamnose 704:730 the residue 3OMe-L-rhamnose 704:730 The one difference is that the residue 3OMe-L-rhamnose is acetylated at the O-2 position in a non-stoichiometric fashion. 28331984 1 1 theme host 144:147 arg1 machinery 149:157 their host machinery 138:157 their host machinery 138:157 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 0 2 from Structure 0:8 arg1 NE-JV-1 48:54 the chlorovirus NE-JV-1 32:54 the chlorovirus NE-JV-1 32:54 Structure of the N-glycans from the chlorovirus NE-JV-1. 28331984 0 3 from NE-JV-1 48:54 arg1 Structure 0:8 Structure 0:8 Structure of the N-glycans from the chlorovirus NE-JV-1 0:54 Structure of the N-glycans from the chlorovirus NE-JV-1. 28331984 0 3 from NE-JV-1 48:54 arg1 N-glycans 17:25 the N-glycans 13:25 the N-glycans from the chlorovirus NE-JV-1 13:54 Structure of the N-glycans from the chlorovirus NE-JV-1. 28331984 1 4 gly glycosylate 162:172 arg1 proteins 180:187 their proteins 174:187 their proteins 174:187 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 2 5 theme capsid 275:280 arg1 proteins 282:289 their capsid proteins 269:289 their capsid proteins 269:289 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 5 6 theme core 618:621 arg1 region 623:628 a core region 616:628 a core region that is conserved in all of the chloroviruses 616:674 Spectroscopic analyses established that this N-glycan consists of a core region that is conserved in all of the chloroviruses. 28331984 0 7 theme chlorovirus 36:46 arg1 NE-JV-1 48:54 the chlorovirus NE-JV-1 32:54 the chlorovirus NE-JV-1 32:54 Structure of the N-glycans from the chlorovirus NE-JV-1. 28331984 1 8 theme recent 70:75 arg1 studies 77:83 recent studies 70:83 recent studies 70:83 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 4 9 theme Micractinium 525:536 arg1 host 488:491 The host 484:491 The host for NE-JV-1 484:503 The host for NE-JV-1 is the zoochlorella Micractinium conductrix. 28331984 4 9 theme Micractinium 525:536 arg1 conductrix 538:547 the zoochlorella Micractinium conductrix 508:547 the zoochlorella Micractinium conductrix 508:547 The host for NE-JV-1 is the zoochlorella Micractinium conductrix. 28331984 6 10 theme non-stoichiometric 771:788 arg1 fashion 790:796 a non-stoichiometric fashion 769:796 a non-stoichiometric fashion 769:796 The one difference is that the residue 3OMe-L-rhamnose is acetylated at the O-2 position in a non-stoichiometric fashion. 28331984 2 11 theme N-linked 310:317 arg1 oligosaccharides 319:334 N-linked oligosaccharides 310:334 N-linked oligosaccharides with unique features 310:355 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 3 12 theme unusual 423:429 arg1 NE-JV-1 444:450 NE-JV-1 444:450 NE-JV-1 444:450 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 3 12 theme unusual 423:429 arg1 chlorovirus 431:441 an unusual chlorovirus 420:441 an unusual chlorovirus 420:441 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 3 13 theme chlorovirus 431:441 arg1 structure 407:415 the N-glycan structure 394:415 the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group 394:481 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 0 14 theme N-glycans 17:25 arg1 Structure 0:8 Structure 0:8 Structure of the N-glycans from the chlorovirus NE-JV-1 0:54 Structure of the N-glycans from the chlorovirus NE-JV-1. 28331984 2 15 with oligosaccharides 319:334 arg1 features 348:355 unique features 341:355 unique features 341:355 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 3 16 theme N-glycan 398:405 arg1 structure 407:415 the N-glycan structure 394:415 the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group 394:481 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 6 17 theme O-2 753:755 arg1 position 757:764 the O-2 position 749:764 the O-2 position 749:764 The one difference is that the residue 3OMe-L-rhamnose is acetylated at the O-2 position in a non-stoichiometric fashion. 28331984 3 18 theme structure 407:415 arg1 elucidation 379:389 the elucidation 375:389 the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group 375:481 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 2 19 link N-linked 310:317 arg1 oligosaccharides 319:334 N-linked oligosaccharides 310:334 N-linked oligosaccharides with unique features 310:355 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 2 20 theme several 211:217 arg1 Chloroviruses 190:202 Chloroviruses 190:202 Chloroviruses 190:202 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 2 20 theme several 211:217 arg1 genes 219:223 several genes 211:223 several genes involved in glycan biosynthesis 211:255 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 5 21 theme Spectroscopic 550:562 arg1 analyses 564:571 Spectroscopic analyses 550:571 Spectroscopic analyses 550:571 Spectroscopic analyses established that this N-glycan consists of a core region that is conserved in all of the chloroviruses. 28331984 2 22 theme unique 341:346 arg1 features 348:355 unique features 341:355 unique features 341:355 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 28331984 4 23 theme zoochlorella 512:523 arg1 host 488:491 The host 484:491 The host for NE-JV-1 484:503 The host for NE-JV-1 is the zoochlorella Micractinium conductrix. 28331984 4 23 theme zoochlorella 512:523 arg1 conductrix 538:547 the zoochlorella Micractinium conductrix 508:547 the zoochlorella Micractinium conductrix 508:547 The host for NE-JV-1 is the zoochlorella Micractinium conductrix. 28331984 3 24 theme Pbi 473:475 arg1 group 477:481 the Pbi group 469:481 the Pbi group 469:481 Here we describe the elucidation of the N-glycan structure of an unusual chlorovirus, NE-JV-1, that belongs to the Pbi group. 28331984 1 25 from studies 77:83 arg1 breaking 89:96 breaking 89:96 breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins 89:187 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 1 25 from studies 77:83 arg1 Results 57:63 Results 57:63 Results from recent studies 57:83 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 1 26 dep breaking 89:96 arg1 paradigm 102:109 the paradigm 98:109 breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins 89:187 Results from recent studies are breaking the paradigm that all viruses depend on their host machinery to glycosylate their proteins. 28331984 2 27 theme glycan 237:242 arg1 biosynthesis 244:255 glycan biosynthesis 237:255 glycan biosynthesis 237:255 Chloroviruses encode several genes involved in glycan biosynthesis and some of their capsid proteins are decorated with N-linked oligosaccharides with unique features. 27743354 4 0 theme ability 571:577 arg1 importance 525:534 the importance 521:534 the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease 521:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 1 1 gly glycosylated 83:94 arg1 half 58:61 More than half 48:61 More than half of all proteins 48:77 More than half of all proteins are glycosylated. 27743354 2 2 theme attached 101:108 arg1 glycans 110:116 The attached glycans 97:116 The attached glycans 97:116 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 4 3 theme proteins 618:625 arg1 phosphorylation 599:613 phosphorylation 599:613 phosphorylation of proteins 599:625 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 3 4 from emphasis 375:382 arg1 regulation 387:396 regulation 387:396 regulation of protein half-life and modulation of protein function by alternative glycosylation 387:481 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 5 theme structural 150:159 arg1 properties 176:185 important structural and functional properties 140:185 important structural and functional properties 140:185 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 3 6 theme function 445:452 arg1 half-life 409:417 protein half-life 401:417 protein half-life 401:417 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 6 theme function 445:452 arg1 modulation 423:432 modulation 423:432 modulation of protein function by alternative glycosylation 423:481 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 7 theme proteins 360:367 arg1 function 348:355 function 348:355 function 348:355 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 7 theme proteins 360:367 arg1 structure 334:342 structure 334:342 structure 334:342 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 8 theme important 140:148 arg1 properties 176:185 important structural and functional properties 140:185 important structural and functional properties 140:185 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 1 9 theme proteins 70:77 arg1 half 58:61 More than half 48:61 More than half of all proteins 48:77 More than half of all proteins are glycosylated. 27743354 2 10 with proteins 126:133 arg1 properties 176:185 important structural and functional properties 140:185 important structural and functional properties 140:185 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 2 10 with proteins 126:133 arg1 parts 198:202 glycan parts 191:202 glycan parts of glycoproteins 191:219 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 0 11 theme Ubiquitous 0:9 arg1 Importance 11:20 Ubiquitous Importance 0:20 Ubiquitous Importance of Protein Glycosylation 0:45 Ubiquitous Importance of Protein Glycosylation. 27743354 4 12 from disease 661:667 arg1 importance 525:534 the importance 521:534 the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease 521:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 1 13 theme than 53:56 arg1 half 58:61 More than half 48:61 More than half of all proteins 48:77 More than half of all proteins are glycosylated. 27743354 0 14 theme Protein 25:31 arg1 Glycosylation 33:45 Protein Glycosylation 25:45 Protein Glycosylation 25:45 Ubiquitous Importance of Protein Glycosylation. 27743354 2 15 theme biological 254:263 arg1 processes 265:273 many key biological processes 245:273 many key biological processes 245:273 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 4 16 from interactions 553:564 arg1 disease 661:667 disease 661:667 disease 661:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 17 theme glycans 650:656 arg1 interactions 553:564 glycan-lectin interactions 539:564 glycan-lectin interactions 539:564 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 17 theme glycans 650:656 arg1 ability 571:577 the ability 567:577 the ability of glycans to block phosphorylation of proteins 567:625 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 17 theme glycans 650:656 arg1 importance 636:645 the importance 632:645 the importance of glycans in disease 632:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 18 from ability 571:577 arg1 disease 661:667 disease 661:667 disease 661:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 3 19 dep structure 334:342 arg1 the 330:332 the 330:332 the 330:332 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 20 theme glycan 191:196 arg1 parts 198:202 glycan parts 191:202 glycan parts of glycoproteins 191:219 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 4 21 from importance 525:534 arg1 disease 661:667 disease 661:667 disease 661:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 22 theme glycans 582:588 arg1 interactions 553:564 glycan-lectin interactions 539:564 glycan-lectin interactions 539:564 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 22 theme glycans 582:588 arg1 ability 571:577 the ability 567:577 the ability of glycans to block phosphorylation of proteins 567:625 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 4 22 theme glycans 582:588 arg1 importance 636:645 the importance 632:645 the importance of glycans in disease 632:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 2 23 theme many 245:248 arg1 processes 265:273 many key biological processes 245:273 many key biological processes 245:273 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 3 24 theme protein 437:443 arg1 function 445:452 protein function 437:452 protein function 437:452 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 25 theme alternative 457:467 arg1 glycosylation 469:481 alternative glycosylation 457:481 alternative glycosylation 457:481 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 0 26 theme Glycosylation 33:45 arg1 Importance 11:20 Ubiquitous Importance 0:20 Ubiquitous Importance of Protein Glycosylation 0:45 Ubiquitous Importance of Protein Glycosylation. 27743354 4 27 theme interactions 553:564 arg1 importance 525:534 the importance 521:534 the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease 521:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 3 28 theme modulation 423:432 arg1 regulation 387:396 regulation 387:396 regulation of protein half-life and modulation of protein function by alternative glycosylation 387:481 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 29 theme protein 401:407 arg1 half-life 409:417 protein half-life 401:417 protein half-life 401:417 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 4 30 theme glycan-lectin 539:551 arg1 interactions 553:564 glycan-lectin interactions 539:564 glycan-lectin interactions 539:564 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 3 31 theme glycosylation 313:325 arg1 effect 303:308 the effect 299:308 the effect of glycosylation on the structure and function of proteins 299:367 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 32 theme functional 165:174 arg1 properties 176:185 important structural and functional properties 140:185 important structural and functional properties 140:185 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 3 33 theme half-life 409:417 arg1 regulation 387:396 regulation 387:396 regulation of protein half-life and modulation of protein function by alternative glycosylation 387:481 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 34 theme essential 226:234 arg1 roles 236:240 essential roles 226:240 essential roles 226:240 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 2 35 gly glycoproteins 207:219 arg1 glycoproteins 207:219 glycoproteins 207:219 glycoproteins 207:219 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 2 36 dep provide 118:124 arg1 have 221:224 have 221:224 provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes 118:273 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 2 37 theme glycoproteins 207:219 arg1 properties 176:185 important structural and functional properties 140:185 important structural and functional properties 140:185 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 2 37 theme glycoproteins 207:219 arg1 parts 198:202 glycan parts 191:202 glycan parts of glycoproteins 191:219 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 27743354 3 38 from effect 303:308 arg1 function 348:355 function 348:355 function 348:355 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 3 38 from effect 303:308 arg1 structure 334:342 structure 334:342 structure 334:342 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 4 39 from importance 636:645 arg1 disease 661:667 disease 661:667 disease 661:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 1 40 theme More 48:51 arg1 half 58:61 More than half 48:61 More than half of all proteins 48:77 More than half of all proteins are glycosylated. 27743354 4 41 theme importance 636:645 arg1 importance 525:534 the importance 521:534 the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease 521:667 In addition, this chapter highlights the importance of glycan-lectin interactions, the ability of glycans to block phosphorylation of proteins, and the importance of glycans in disease. 27743354 3 42 gly glycosylation 313:325 arg1 proteins 360:367 proteins 360:367 proteins 360:367 This chapter describes the effect of glycosylation on the structure and function of proteins, with emphasis on regulation of protein half-life and modulation of protein function by alternative glycosylation. 27743354 2 43 theme key 250:252 arg1 processes 265:273 many key biological processes 245:273 many key biological processes 245:273 The attached glycans provide proteins with important structural and functional properties and glycan parts of glycoproteins have essential roles in many key biological processes. 26599081 0 0 theme Exogenous 80:88 arg1 Oligosaccharide 90:104 Exogenous Oligosaccharide 80:104 Exogenous Oligosaccharide 80:104 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 5 1 contain possess 876:882 arg1 species 835:841 both species 830:841 both species of mycoplasma 830:855 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 1 contain possess 876:882 arg2 glycans 893:899 N-linked glycans 884:899 N-linked glycans 884:899 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 3 2 theme Mycoplasma 469:478 arg1 arthritidis 480:490 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 arthritidis 480:490 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 2 theme Mycoplasma 469:478 arg1 pathogens 459:467 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 1 3 theme membrane 222:229 arg1 possession 194:203 the possession 190:203 the possession of only a single membrane 190:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 3 theme membrane 222:229 arg1 lack 111:114 The lack 107:114 The lack of a cell wall, flagella, fimbria, and other extracellular appendages 107:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 5 4 theme O-linked 767:774 arg1 glycoproteins 776:788 O-linked glycoproteins 767:788 O-linked glycoproteins 767:788 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 1 5 theme glycoconjugates 321:335 arg1 study 312:316 the study 308:316 the study of glycoconjugates 308:335 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 9 6 theme N-linked 1245:1252 arg1 glycoproteins 1254:1266 N-linked glycoproteins 1245:1266 N-linked glycoproteins 1245:1266 As far as we are aware, N-linked glycoproteins have not been previously described in Gram-positive bacteria, the organisms to which the mycoplasmas are phylogenetically related. 26599081 1 7 theme fimbria 142:148 arg1 possession 194:203 the possession 190:203 the possession of only a single membrane 190:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 7 theme fimbria 142:148 arg1 lack 111:114 The lack 107:114 The lack of a cell wall, flagella, fimbria, and other extracellular appendages 107:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 3 8 gly glycoproteins 632:644 arg1 glycoproteins 632:644 glycoproteins 632:644 glycoproteins 632:644 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 6 9 gly glycosylation 950:962 arg2 sites 985:989 numerous sites 976:989 numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity 976:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 5 10 theme N-linked 884:891 arg1 glycans 893:899 N-linked glycans 884:899 N-linked glycans 884:899 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 4 11 dep glycosylated 692:703 arg1 arthritidis 680:690 M. arthritidis 677:690 M. arthritidis 677:690 Previously, it was shown that M. arthritidis glycosylated surface lipoproteins through O-linkage. 26599081 6 12 theme surface-exposed 994:1008 arg1 lipoproteins 1010:1021 surface-exposed lipoproteins 994:1021 surface-exposed lipoproteins with no apparent amino acid sequence specificity 994:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 0 13 theme Oligosaccharide 90:104 arg1 Glycosylation 23:35 General N-and O-Linked Glycosylation 0:35 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas 0:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 0 13 theme Oligosaccharide 90:104 arg1 Role 72:75 Role 72:75 Role of Exogenous Oligosaccharide 72:104 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 5 14 theme M. 814:815 arg1 pulmonis 817:824 M. pulmonis 814:824 M. pulmonis 814:824 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 10 15 theme cell 1441:1444 arg1 surface 1446:1452 the mycoplasma cell surface 1426:1452 the mycoplasma cell surface 1426:1452 The findings indicate that the mycoplasma cell surface is heavily glycosylated with implications for the modulation of mycoplasma-host interactions. 26599081 4 16 theme M. 677:678 arg1 arthritidis 680:690 M. arthritidis 677:690 M. arthritidis 677:690 Previously, it was shown that M. arthritidis glycosylated surface lipoproteins through O-linkage. 26599081 4 17 theme surface 705:711 arg1 lipoproteins 713:724 M. arthritidis glycosylated surface lipoproteins 677:724 M. arthritidis glycosylated surface lipoproteins 677:724 Previously, it was shown that M. arthritidis glycosylated surface lipoproteins through O-linkage. 26599081 10 18 theme mycoplasma 1430:1439 arg1 surface 1446:1452 the mycoplasma cell surface 1426:1452 the mycoplasma cell surface 1426:1452 The findings indicate that the mycoplasma cell surface is heavily glycosylated with implications for the modulation of mycoplasma-host interactions. 26599081 9 19 link N-linked 1245:1252 arg1 glycoproteins 1254:1266 N-linked glycoproteins 1245:1266 N-linked glycoproteins 1245:1266 As far as we are aware, N-linked glycoproteins have not been previously described in Gram-positive bacteria, the organisms to which the mycoplasmas are phylogenetically related. 26599081 2 20 theme kb 377:378 arg1 genomes 356:362 genomes 356:362 genomes of about 800 kb 356:378 Most species have genomes of about 800 kb and code for few proteins predicted to have a role in glycobiology. 26599081 2 21 contain have 351:354 arg2 genomes 356:362 genomes 356:362 genomes of about 800 kb 356:378 Most species have genomes of about 800 kb and code for few proteins predicted to have a role in glycobiology. 26599081 2 21 contain have 351:354 arg1 species 343:349 Most species 338:349 Most species 338:349 Most species have genomes of about 800 kb and code for few proteins predicted to have a role in glycobiology. 26599081 3 22 dep pathogens 459:467 arg1 arthritidis 480:490 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 arthritidis 480:490 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 22 dep pathogens 459:467 arg1 pulmonis 507:514 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 pulmonis 507:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 22 dep pathogens 459:467 arg1 pathogens 459:467 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 7 23 gly glycosylated 1124:1135 arg1 lipoproteins 1077:1088 The lipoproteins 1073:1088 The lipoproteins of Mycoplasma pneumoniae 1073:1113 The lipoproteins of Mycoplasma pneumoniae also are glycosylated. 26599081 0 24 theme N-and 8:12 arg1 Glycosylation 23:35 General N-and O-Linked Glycosylation 0:35 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas 0:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 3 25 theme Mycoplasma 496:505 arg1 pulmonis 507:514 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 pulmonis 507:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 25 theme Mycoplasma 496:505 arg1 pathogens 459:467 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 10 26 theme interactions 1534:1545 arg1 modulation 1504:1513 the modulation 1500:1513 the modulation of mycoplasma-host interactions 1500:1545 The findings indicate that the mycoplasma cell surface is heavily glycosylated with implications for the modulation of mycoplasma-host interactions. 26599081 8 27 theme glycosidic 1173:1182 arg1 linkages 1184:1191 the glycosidic linkages 1169:1191 the glycosidic linkages from host oligosaccharides 1169:1218 Glycosylation was dependent on the glycosidic linkages from host oligosaccharides. 26599081 5 28 theme glutamine 931:939 arg1 glutamine 931:939 glutamine 931:939 glutamine 931:939 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 28 theme glutamine 931:939 arg1 asparagine 916:925 asparagine 916:925 asparagine 916:925 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 28 theme glutamine 931:939 arg1 residues 904:911 residues 904:911 residues of asparagine and glutamine 904:939 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 1 29 theme other 155:159 arg1 appendages 175:184 other extracellular appendages 155:184 other extracellular appendages 155:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 0 30 theme General 0:6 arg1 Glycosylation 23:35 General N-and O-Linked Glycosylation 0:35 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas 0:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 10 31 theme mycoplasma-host 1518:1532 arg1 interactions 1534:1545 mycoplasma-host interactions 1518:1545 mycoplasma-host interactions 1518:1545 The findings indicate that the mycoplasma cell surface is heavily glycosylated with implications for the modulation of mycoplasma-host interactions. 26599081 5 32 located found 805:809 arg1 study 760:764 the current study 748:764 the current study 748:764 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 32 located found 805:809 arg1 pulmonis 817:824 M. pulmonis 814:824 M. pulmonis 814:824 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 32 located found 805:809 arg2 glycoproteins 776:788 O-linked glycoproteins 767:788 O-linked glycoproteins 767:788 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 33 link O-linked 767:774 arg1 glycoproteins 776:788 O-linked glycoproteins 767:788 O-linked glycoproteins 767:788 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 1 34 theme extracellular 161:173 arg1 appendages 175:184 other extracellular appendages 155:184 other extracellular appendages 155:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 35 theme simplistic 267:276 arg1 mycoplasmas 242:252 the mycoplasmas 238:252 the mycoplasmas structurally simplistic 238:276 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 4 36 gly glycosylated 692:703 arg1 lipoproteins 713:724 M. arthritidis glycosylated surface lipoproteins 677:724 M. arthritidis glycosylated surface lipoproteins 677:724 Previously, it was shown that M. arthritidis glycosylated surface lipoproteins through O-linkage. 26599081 3 37 theme single 528:533 arg1 gene 535:538 only a single gene 521:538 only a single gene annotated as coding for a glycosyltransferase 521:584 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 1 38 theme appendages 175:184 arg1 possession 194:203 the possession 190:203 the possession of only a single membrane 190:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 38 theme appendages 175:184 arg1 lack 111:114 The lack 107:114 The lack of a cell wall, flagella, fimbria, and other extracellular appendages 107:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 0 39 theme O-Linked 14:21 arg1 Glycosylation 23:35 General N-and O-Linked Glycosylation 0:35 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas 0:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 7 40 theme pneumoniae 1104:1113 arg1 lipoproteins 1077:1088 The lipoproteins 1073:1088 The lipoproteins of Mycoplasma pneumoniae 1073:1113 The lipoproteins of Mycoplasma pneumoniae also are glycosylated. 26599081 2 41 theme Most 338:341 arg1 species 343:349 Most species 338:349 Most species 338:349 Most species have genomes of about 800 kb and code for few proteins predicted to have a role in glycobiology. 26599081 10 42 gly glycosylated 1465:1476 arg1 surface 1446:1452 the mycoplasma cell surface 1426:1452 the mycoplasma cell surface 1426:1452 The findings indicate that the mycoplasma cell surface is heavily glycosylated with implications for the modulation of mycoplasma-host interactions. 26599081 0 43 theme Lipoproteins 40:51 arg1 Glycosylation 23:35 General N-and O-Linked Glycosylation 0:35 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas 0:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 0 43 theme Lipoproteins 40:51 arg1 Role 72:75 Role 72:75 Role of Exogenous Oligosaccharide 72:104 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 1 44 theme ideal 282:286 arg1 organisms 294:302 ideal model organisms 282:302 ideal model organisms 282:302 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 6 45 from sites 985:989 arg1 lipoproteins 1010:1021 surface-exposed lipoproteins 994:1021 surface-exposed lipoproteins with no apparent amino acid sequence specificity 994:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 1 46 theme model 288:292 arg1 organisms 294:302 ideal model organisms 282:302 ideal model organisms 282:302 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 5 47 link N-linked 884:891 arg1 glycans 893:899 N-linked glycans 884:899 N-linked glycans 884:899 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 0 48 from Role 72:75 arg1 Mycoplasmas 56:66 Mycoplasmas 56:66 Mycoplasmas 56:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 9 49 theme Gram-positive 1306:1318 arg1 organisms 1334:1342 the organisms 1330:1342 the organisms to which the mycoplasmas are phylogenetically related 1330:1396 As far as we are aware, N-linked glycoproteins have not been previously described in Gram-positive bacteria, the organisms to which the mycoplasmas are phylogenetically related. 26599081 9 49 theme Gram-positive 1306:1318 arg1 bacteria 1320:1327 Gram-positive bacteria 1306:1327 Gram-positive bacteria 1306:1327 As far as we are aware, N-linked glycoproteins have not been previously described in Gram-positive bacteria, the organisms to which the mycoplasmas are phylogenetically related. 26599081 5 50 theme mycoplasma 846:855 arg1 species 835:841 both species 830:841 both species of mycoplasma 830:855 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 9 51 gly glycoproteins 1254:1266 arg1 glycoproteins 1254:1266 N-linked glycoproteins 1245:1266 N-linked glycoproteins 1245:1266 As far as we are aware, N-linked glycoproteins have not been previously described in Gram-positive bacteria, the organisms to which the mycoplasmas are phylogenetically related. 26599081 3 52 contain have 516:519 arg1 arthritidis 480:490 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 arthritidis 480:490 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 52 contain have 516:519 arg2 gene 535:538 only a single gene 521:538 only a single gene annotated as coding for a glycosyltransferase 521:584 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 52 contain have 516:519 arg1 pathogens 459:467 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 52 contain have 516:519 arg1 pulmonis 507:514 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 pulmonis 507:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 6 53 theme numerous 976:983 arg1 sites 985:989 numerous sites 976:989 numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity 976:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 3 54 theme murine 452:457 arg1 arthritidis 480:490 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 arthritidis 480:490 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 54 theme murine 452:457 arg1 pulmonis 507:514 Mycoplasma arthritidis and Mycoplasma pulmonis 469:514 pulmonis 507:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 3 54 theme murine 452:457 arg1 pathogens 459:467 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis 448:514 The murine pathogens Mycoplasma arthritidis and Mycoplasma pulmonis have only a single gene annotated as coding for a glycosyltransferase but synthesize glycolipid, polysaccharide and glycoproteins. 26599081 7 55 theme Mycoplasma 1093:1102 arg1 pneumoniae 1104:1113 Mycoplasma pneumoniae 1093:1113 Mycoplasma pneumoniae 1093:1113 The lipoproteins of Mycoplasma pneumoniae also are glycosylated. 26599081 0 56 gly Glycosylation 23:35 arg1 Lipoproteins 40:51 Lipoproteins 40:51 Lipoproteins 40:51 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 0 56 gly Glycosylation 23:35 arg1 Oligosaccharide 90:104 Exogenous Oligosaccharide 80:104 Exogenous Oligosaccharide 80:104 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 4 57 theme glycosylated 692:703 arg1 lipoproteins 713:724 M. arthritidis glycosylated surface lipoproteins 677:724 M. arthritidis glycosylated surface lipoproteins 677:724 Previously, it was shown that M. arthritidis glycosylated surface lipoproteins through O-linkage. 26599081 8 58 theme host 1198:1201 arg1 oligosaccharides 1203:1218 host oligosaccharides 1198:1218 host oligosaccharides 1198:1218 Glycosylation was dependent on the glycosidic linkages from host oligosaccharides. 26599081 6 59 theme amino 1040:1044 arg1 specificity 1060:1070 no apparent amino acid sequence specificity 1028:1070 no apparent amino acid sequence specificity 1028:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 1 60 theme cell 121:124 arg1 wall 126:129 a cell wall 119:129 a cell wall 119:129 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 6 61 theme acid 1046:1049 arg1 specificity 1060:1070 no apparent amino acid sequence specificity 1028:1070 no apparent amino acid sequence specificity 1028:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 8 62 from oligosaccharides 1203:1218 arg1 linkages 1184:1191 the glycosidic linkages 1169:1191 the glycosidic linkages from host oligosaccharides 1169:1218 Glycosylation was dependent on the glycosidic linkages from host oligosaccharides. 26599081 5 63 gly glycoproteins 776:788 arg1 glycoproteins 776:788 O-linked glycoproteins 767:788 O-linked glycoproteins 767:788 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 1 64 theme wall 126:129 arg1 possession 194:203 the possession 190:203 the possession of only a single membrane 190:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 64 theme wall 126:129 arg1 lack 111:114 The lack 107:114 The lack of a cell wall, flagella, fimbria, and other extracellular appendages 107:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 5 65 theme asparagine 916:925 arg1 glutamine 931:939 glutamine 931:939 glutamine 931:939 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 65 theme asparagine 916:925 arg1 asparagine 916:925 asparagine 916:925 asparagine 916:925 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 65 theme asparagine 916:925 arg1 residues 904:911 residues 904:911 residues of asparagine and glutamine 904:939 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 5 66 theme current 752:758 arg1 study 760:764 the current study 748:764 the current study 748:764 In the current study, O-linked glycoproteins were similarly found in M. pulmonis and both species of mycoplasma were found to also possess N-linked glycans at residues of asparagine and glutamine. 26599081 6 67 theme sequence 1051:1058 arg1 specificity 1060:1070 no apparent amino acid sequence specificity 1028:1070 no apparent amino acid sequence specificity 1028:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 2 68 theme few 393:395 arg1 proteins 397:404 few proteins 393:404 few proteins predicted to have a role in glycobiology 393:445 Most species have genomes of about 800 kb and code for few proteins predicted to have a role in glycobiology. 26599081 6 69 theme apparent 1031:1038 arg1 specificity 1060:1070 no apparent amino acid sequence specificity 1028:1070 no apparent amino acid sequence specificity 1028:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 0 70 from Glycosylation 23:35 arg1 Mycoplasmas 56:66 Mycoplasmas 56:66 Mycoplasmas 56:66 General N-and O-Linked Glycosylation of Lipoproteins in Mycoplasmas and Role of Exogenous Oligosaccharide. 26599081 6 71 with lipoproteins 1010:1021 arg1 specificity 1060:1070 no apparent amino acid sequence specificity 1028:1070 no apparent amino acid sequence specificity 1028:1070 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 1 72 theme flagella 132:139 arg1 possession 194:203 the possession 190:203 the possession of only a single membrane 190:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 1 72 theme flagella 132:139 arg1 lack 111:114 The lack 107:114 The lack of a cell wall, flagella, fimbria, and other extracellular appendages 107:184 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 26599081 6 73 theme Protein 942:948 arg1 glycosylation 950:962 Protein glycosylation 942:962 Protein glycosylation 942:962 Protein glycosylation occurred at numerous sites on surface-exposed lipoproteins with no apparent amino acid sequence specificity. 26599081 1 74 theme single 215:220 arg1 membrane 222:229 only a single membrane 208:229 only a single membrane 208:229 The lack of a cell wall, flagella, fimbria, and other extracellular appendages and the possession of only a single membrane render the mycoplasmas structurally simplistic and ideal model organisms for the study of glycoconjugates. 28973586 11 0 theme auxin 1547:1551 arg1 types 1553:1557 auxin types 1547:1557 auxin types 1547:1557 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 10 1 theme higher 1424:1429 arg1 stability 1431:1439 higher stability 1424:1439 higher stability 1424:1439 NAA bound protein folding was more similar to the crystal structure showing higher stability compared to that of IAA bound. 28973586 9 2 theme egress 1331:1336 arg1 entrance 1319:1326 entrance 1319:1326 entrance 1319:1326 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 2 theme egress 1331:1336 arg1 site 1311:1314 the putative site 1298:1314 the putative site of entrance or egress of auxin 1298:1345 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 2 theme egress 1331:1336 arg1 egress 1331:1336 egress 1331:1336 egress 1331:1336 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 0 3 from role 4:7 arg1 interaction 58:68 the interaction 54:68 the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins 54:124 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 5 4 theme molecular 590:598 arg1 simulations 609:619 molecular dynamics simulations 590:619 molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 590:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 5 theme structural 533:542 arg1 biology 544:550 ABP1 structural biology 528:550 ABP1 structural biology 528:550 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 7 6 from regions 931:937 arg1 ABP1 942:945 ABP1 942:945 ABP1 of both species 942:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 10 7 theme crystal 1398:1404 arg1 structure 1406:1414 the crystal structure 1394:1414 the crystal structure showing higher stability compared to that of IAA bound 1394:1469 NAA bound protein folding was more similar to the crystal structure showing higher stability compared to that of IAA bound. 28973586 7 8 from structure 1017:1025 arg1 maize 1030:1034 maize 1030:1034 maize 1030:1034 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 0 9 theme Protein 87:93 arg1 interaction 58:68 the interaction 54:68 the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins 54:124 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 5 10 theme dynamics 600:607 arg1 simulations 609:619 molecular dynamics simulations 590:619 molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 590:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 0 11 with interaction 58:68 arg1 auxins 119:124 different auxins 109:124 different auxins 109:124 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 2 12 theme 1-naphthalene 241:253 arg1 acid 262:265 1-naphthalene acetic acid 241:265 1-naphthalene acetic acid (NAA) 241:271 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 12 theme 1-naphthalene 241:253 arg1 form 288:291 a synthetic form 276:291 a synthetic form 276:291 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 12 theme 1-naphthalene 241:253 arg1 NAA 268:270 NAA 268:270 NAA 268:270 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 9 13 theme putative 1302:1309 arg1 entrance 1319:1326 entrance 1319:1326 entrance 1319:1326 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 13 theme putative 1302:1309 arg1 site 1311:1314 the putative site 1298:1314 the putative site of entrance or egress of auxin 1298:1345 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 13 theme putative 1302:1309 arg1 egress 1331:1336 egress 1331:1336 egress 1331:1336 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 14 theme auxin 1341:1345 arg1 entrance 1319:1326 entrance 1319:1326 entrance 1319:1326 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 14 theme auxin 1341:1345 arg1 egress 1331:1336 egress 1331:1336 egress 1331:1336 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 0 15 theme Auxin-Binding 73:85 arg1 Protein 87:93 Auxin-Binding Protein 1 73:95 Auxin-Binding Protein 1 (ABP1) 73:102 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 0 15 theme Auxin-Binding 73:85 arg1 ABP1 98:101 ABP1 98:101 ABP1 98:101 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 5 16 theme maize 689:693 arg1 thaliana 711:718 maize and Arabidopsis thaliana 689:718 thaliana 711:718 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 6 17 theme stabilization 846:858 arg1 effect 860:865 such stabilization effect 841:865 such stabilization effect 841:865 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 10 18 theme protein 1358:1364 arg1 folding 1366:1372 protein folding 1358:1372 protein folding 1358:1372 NAA bound protein folding was more similar to the crystal structure showing higher stability compared to that of IAA bound. 28973586 6 19 theme such 841:844 arg1 effect 860:865 such stabilization effect 841:865 such stabilization effect 841:865 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 5 20 theme further 509:515 arg1 insight 517:523 further insight 509:523 further insight on ABP1 structural biology 509:550 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 8 21 theme monomeric 1116:1124 arg1 simulations 1126:1136 the monomeric simulations 1112:1136 the monomeric simulations 1112:1136 In Arabidopsis, the helix appeared more stable, being preserved in most of the monomeric simulations and unfolding when the protein was in the dimeric form. 28973586 7 22 from ABP1 942:945 arg1 α-helix 909:915 The α-helix 905:915 The α-helix of C-terminal regions in ABP1 of both species 905:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 4 23 theme ABP1 427:430 arg1 structure 432:440 ABP1 structure 427:440 ABP1 structure 427:440 ABP1 structure was previously established from maize at 1.9 Å resolution. 28973586 11 24 theme structural 1486:1495 arg1 differences 1497:1507 The molecular structural differences 1472:1507 The molecular structural differences of ABP1 found between the species and auxin types 1472:1557 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 1 25 theme plant 149:153 arg1 growth 155:160 plant growth 149:160 plant growth 149:160 Auxin is critical for plant growth and development. 28973586 5 26 theme oligomeric 651:660 arg1 proteins 675:682 the oligomeric glycosylated proteins 647:682 the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 647:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 6 27 theme N-terminal 886:895 arg1 region 897:902 the N-terminal region 882:902 the N-terminal region 882:902 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 5 28 theme proteins 675:682 arg1 models 637:642 the complete models 624:642 the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 624:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 8 29 theme dimeric 1180:1186 arg1 form 1188:1191 the dimeric form 1176:1191 the dimeric form 1176:1191 In Arabidopsis, the helix appeared more stable, being preserved in most of the monomeric simulations and unfolding when the protein was in the dimeric form. 28973586 0 30 theme Zn2+ 12:15 arg1 role 4:7 The role 0:7 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 0:125 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 11 31 theme auxin-binding 1578:1590 arg1 protein 1592:1598 this auxin-binding protein 1573:1598 this auxin-binding protein 1573:1598 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 5 32 theme ABP1 528:531 arg1 biology 544:550 ABP1 structural biology 528:550 ABP1 structural biology 528:550 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 6 33 theme Zn2+ 759:762 arg1 coordination 764:775 Zn2+ coordination 759:775 Zn2+ coordination 759:775 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 3 34 theme nontranscriptional 392:409 arg1 responses 416:424 nontranscriptional cell responses 392:424 nontranscriptional cell responses 392:424 Auxin-Binding Protein 1 (ABP1) specifically binds auxins, presumably playing roles as receptor in nontranscriptional cell responses. 28973586 11 35 theme auxin 1669:1673 arg1 binding 1680:1686 auxin type binding 1669:1686 auxin type binding 1669:1686 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 7 36 theme extended 1008:1015 arg1 structure 1017:1025 a more extended structure 1001:1025 a more extended structure in maize 1001:1034 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 2 37 theme acetic 255:260 arg1 acid 262:265 1-naphthalene acetic acid 241:265 1-naphthalene acetic acid (NAA) 241:271 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 37 theme acetic 255:260 arg1 form 288:291 a synthetic form 276:291 a synthetic form 276:291 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 37 theme acetic 255:260 arg1 NAA 268:270 NAA 268:270 NAA 268:270 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 3 38 theme cell 411:414 arg1 responses 416:424 nontranscriptional cell responses 392:424 nontranscriptional cell responses 392:424 Auxin-Binding Protein 1 (ABP1) specifically binds auxins, presumably playing roles as receptor in nontranscriptional cell responses. 28973586 0 39 theme dimerization 18:29 arg1 role 4:7 The role 0:7 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 0:125 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 0 40 theme different 109:117 arg1 auxins 119:124 different auxins 109:124 different auxins 109:124 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 5 41 theme glycosylated 662:673 arg1 proteins 675:682 the oligomeric glycosylated proteins 647:682 the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 647:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 11 42 theme functional 1606:1615 arg1 specificities 1617:1629 functional specificities 1606:1629 functional specificities 1606:1629 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 3 43 from receptor 380:387 arg1 responses 416:424 nontranscriptional cell responses 392:424 nontranscriptional cell responses 392:424 Auxin-Binding Protein 1 (ABP1) specifically binds auxins, presumably playing roles as receptor in nontranscriptional cell responses. 28973586 6 44 located located 871:877 arg2 most 833:836 most 833:836 most 833:836 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 6 44 located located 871:877 arg2 effect 860:865 such stabilization effect 841:865 such stabilization effect 841:865 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 6 44 located located 871:877 arg1 region 897:902 the N-terminal region 882:902 the N-terminal region 882:902 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 0 45 theme N-glycosylation 35:49 arg1 role 4:7 The role 0:7 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 0:125 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 3 46 theme Auxin-Binding 294:306 arg1 ABP1 319:322 ABP1 319:322 ABP1 319:322 Auxin-Binding Protein 1 (ABP1) specifically binds auxins, presumably playing roles as receptor in nontranscriptional cell responses. 28973586 3 46 theme Auxin-Binding 294:306 arg1 Protein 308:314 Auxin-Binding Protein 1 294:316 Auxin-Binding Protein 1 (ABP1) 294:323 Auxin-Binding Protein 1 (ABP1) specifically binds auxins, presumably playing roles as receptor in nontranscriptional cell responses. 28973586 6 47 theme conformational 804:817 arg1 stability 819:827 conformational stability 804:827 conformational stability 804:827 In maize, both Zn2+ coordination and glycosylation promoted conformational stability and most of such stabilization effect was located on the N-terminal region. 28973586 2 48 theme natural 188:194 arg1 auxin 196:200 The main natural auxin 179:200 The main natural auxin 179:200 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 48 theme natural 188:194 arg1 acid 221:224 indole-3-acetic acid 205:224 indole-3-acetic acid (IAA) 205:230 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 9 49 theme glycosylation 1235:1247 arg1 structures 1249:1258 glycosylation structures 1235:1258 glycosylation structures 1235:1258 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 2 50 theme synthetic 278:286 arg1 acid 262:265 1-naphthalene acetic acid 241:265 1-naphthalene acetic acid (NAA) 241:271 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 50 theme synthetic 278:286 arg1 form 288:291 a synthetic form 276:291 a synthetic form 276:291 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 4 51 theme Å 487:487 arg1 resolution 489:498 1.9 Å resolution 483:498 1.9 Å resolution 483:498 ABP1 structure was previously established from maize at 1.9 Å resolution. 28973586 2 52 theme main 183:186 arg1 auxin 196:200 The main natural auxin 179:200 The main natural auxin 179:200 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 52 theme main 183:186 arg1 acid 221:224 indole-3-acetic acid 205:224 indole-3-acetic acid (IAA) 205:230 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 7 53 theme species 955:961 arg1 ABP1 942:945 ABP1 942:945 ABP1 of both species 942:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 0 54 gly N-glycosylation 35:49 arg1 Protein 87:93 Auxin-Binding Protein 1 73:95 Auxin-Binding Protein 1 (ABP1) 73:102 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 0 54 gly N-glycosylation 35:49 arg1 ABP1 98:101 ABP1 98:101 ABP1 98:101 The role of Zn2+, dimerization and N-glycosylation in the interaction of Auxin-Binding Protein 1 (ABP1) with different auxins. 28973586 5 55 theme complete 628:635 arg1 models 637:642 the complete models 624:642 the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 624:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 11 56 theme molecular 1476:1484 arg1 differences 1497:1507 The molecular structural differences 1472:1507 The molecular structural differences of ABP1 found between the species and auxin types 1472:1557 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 7 57 theme regions 931:937 arg1 α-helix 909:915 The α-helix 905:915 The α-helix of C-terminal regions in ABP1 of both species 905:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 5 58 gly glycosylated 662:673 arg1 proteins 675:682 the oligomeric glycosylated proteins 647:682 the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 647:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 59 theme models 637:642 arg1 simulations 609:619 molecular dynamics simulations 590:619 molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 590:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 11 60 theme ABP1 1512:1515 arg1 differences 1497:1507 The molecular structural differences 1472:1507 The molecular structural differences of ABP1 found between the species and auxin types 1472:1557 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 5 61 from insight 517:523 arg1 biology 544:550 ABP1 structural biology 528:550 ABP1 structural biology 528:550 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 62 from models 637:642 arg1 thaliana 711:718 maize and Arabidopsis thaliana 689:718 thaliana 711:718 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 63 theme Arabidopsis 699:709 arg1 thaliana 711:718 maize and Arabidopsis thaliana 689:718 thaliana 711:718 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 64 from thaliana 711:718 arg1 proteins 675:682 the oligomeric glycosylated proteins 647:682 the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 647:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 5 64 from thaliana 711:718 arg1 models 637:642 the complete models 624:642 the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins 624:741 To gain further insight on ABP1 structural biology, this study was carried out employing molecular dynamics simulations of the complete models of the oligomeric glycosylated proteins from maize and Arabidopsis thaliana with or without auxins. 28973586 7 65 theme C-terminal 920:929 arg1 regions 931:937 C-terminal regions 920:937 C-terminal regions in ABP1 of both species 920:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 28973586 9 66 theme entrance 1319:1326 arg1 entrance 1319:1326 entrance 1319:1326 entrance 1319:1326 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 66 theme entrance 1319:1326 arg1 site 1311:1314 the putative site 1298:1314 the putative site of entrance or egress of auxin 1298:1345 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 9 66 theme entrance 1319:1326 arg1 egress 1331:1336 egress 1331:1336 egress 1331:1336 In Arabidopsis ABP1 bound to IAA or NAA, glycosylation structures arranged around the protein, covering the putative site of entrance or egress of auxin. 28973586 11 67 theme type 1675:1678 arg1 binding 1680:1686 auxin type binding 1669:1686 auxin type binding 1669:1686 The molecular structural differences of ABP1 found between the species and auxin types indicate that this auxin-binding protein shows functional specificities in dicots and monocots, as well as in auxin type binding. 28973586 2 68 theme indole-3-acetic 205:219 arg1 auxin 196:200 The main natural auxin 179:200 The main natural auxin 179:200 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 68 theme indole-3-acetic 205:219 arg1 acid 221:224 indole-3-acetic acid 205:224 indole-3-acetic acid (IAA) 205:230 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 2 68 theme indole-3-acetic 205:219 arg1 IAA 227:229 IAA 227:229 IAA 227:229 The main natural auxin is indole-3-acetic acid (IAA), whereas 1-naphthalene acetic acid (NAA) is a synthetic form. 28973586 7 69 from α-helix 909:915 arg1 ABP1 942:945 ABP1 942:945 ABP1 of both species 942:961 The α-helix of C-terminal regions in ABP1 of both species unfolded during simulations, assuming a more extended structure in maize. 25477510 4 0 theme O-glycans 933:941 arg1 presence 915:922 the presence 911:922 the presence of minor O-glycans carrying a xylose residue 911:967 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 6 1 theme KRE2/MNT1 1177:1185 arg1 genes 1194:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 8 2 theme increased 1709:1717 arg1 sensitivity 1719:1729 increased sensitivity 1709:1729 increased sensitivity to SDS, high salt, and high temperature 1709:1769 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 8 3 theme full 1923:1926 arg1 pathogenicity 1928:1940 full pathogenicity 1923:1940 full pathogenicity of C. neoformans 1923:1957 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 3 4 theme O-glycans 527:535 arg1 analysis 511:518 a detailed structural analysis 489:518 a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins 489:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 7 5 theme minor 1557:1561 arg1 O-glycans 1563:1571 minor O-glycans 1557:1571 minor O-glycans containing xylose 1557:1589 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 6 theme third 1510:1514 arg1 residue 1524:1530 the third mannose residue 1506:1530 the third mannose residue 1506:1530 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 8 7 theme neoformans 1948:1957 arg1 integrity 1909:1917 cell integrity 1904:1917 cell integrity 1904:1917 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 8 7 theme neoformans 1948:1957 arg1 pathogenicity 1928:1940 full pathogenicity 1923:1940 full pathogenicity of C. neoformans 1923:1957 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 2 8 theme neoformans 300:309 arg1 mannoproteins 280:292 The cell surface mannoproteins 263:292 The cell surface mannoproteins of C. neoformans 263:309 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 6 9 theme responsible 1249:1259 arg1 α1,2-mannosyltransferase 1224:1247 an α1,2-mannosyltransferase 1221:1247 an α1,2-mannosyltransferase responsible for the addition of the second mannose residue 1221:1306 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 6 9 theme responsible 1249:1259 arg1 KTR3 1122:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 2 10 theme host 342:345 arg1 response 354:361 the host T-cell response 338:361 the host T-cell response 338:361 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 3 11 theme third 794:798 arg1 residue 808:814 the third mannose residue 790:814 the third mannose residue 790:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 0 12 theme human 106:110 arg1 neoformans 142:151 the human pathogenic yeast Cryptococcus neoformans 102:151 the human pathogenic yeast Cryptococcus neoformans 102:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 0 13 theme yeast 123:127 arg1 neoformans 142:151 the human pathogenic yeast Cryptococcus neoformans 102:151 the human pathogenic yeast Cryptococcus neoformans 102:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 8 14 theme cryptococcosis 1820:1833 arg1 model 1811:1815 a mouse model 1803:1815 a mouse model of cryptococcosis 1803:1833 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 1 15 theme immunocompromised 237:253 arg1 humans 255:260 immunocompromised humans 237:260 immunocompromised humans 237:260 Cryptococcus neoformans is an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans. 25477510 0 16 theme neoformans 142:151 arg1 apparatus 89:97 the Golgi apparatus 79:97 the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans 79:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 6 17 theme residue 1300:1306 arg1 addition 1269:1276 the addition 1265:1276 the addition of the second mannose residue 1265:1306 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 5 18 theme neoformans 993:1002 arg1 strains 1011:1017 C. neoformans mutant strains 990:1017 C. neoformans mutant strains 990:1017 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 6 19 theme C. 1108:1109 arg1 α1,2-mannosyltransferase 1224:1247 an α1,2-mannosyltransferase 1221:1247 an α1,2-mannosyltransferase responsible for the addition of the second mannose residue 1221:1306 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 6 19 theme C. 1108:1109 arg1 KTR3 1122:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 6 19 theme C. 1108:1109 arg1 homolog 1137:1143 the only homolog 1128:1143 the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes 1128:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 7 20 theme family 1427:1432 arg1 genes 1434:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 0 21 theme O-linked 59:66 arg1 glycans 68:74 O-linked glycans 59:74 O-linked glycans 59:74 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 2 22 theme surface 272:278 arg1 mannoproteins 280:292 The cell surface mannoproteins 263:292 The cell surface mannoproteins of C. neoformans 263:309 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 4 23 contain carrying 943:950 arg1 O-glycans 933:941 minor O-glycans 927:941 minor O-glycans carrying a xylose residue 927:967 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 4 23 contain carrying 943:950 arg2 residue 961:967 a xylose residue 952:967 a xylose residue 952:967 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 8 24 theme O-glycans 1878:1886 arg1 structure 1865:1873 the extended structure 1852:1873 the extended structure of O-glycans 1852:1886 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 5 25 theme Further 970:976 arg1 analyses 978:985 Further analyses 970:985 Further analyses of C. neoformans mutant strains 970:1017 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 6 26 theme mannose 1292:1298 arg1 residue 1300:1306 the second mannose residue 1281:1306 the second mannose residue 1281:1306 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 0 27 theme Golgi 83:87 arg1 apparatus 89:97 the Golgi apparatus 79:97 the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans 79:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 0 28 from pathway 48:54 arg1 apparatus 89:97 the Golgi apparatus 79:97 the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans 79:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 4 29 theme O-glycan 835:842 arg1 profiles 844:851 the O-glycan profiles 831:851 the O-glycan profiles from wild-type and uxs1Δ mutant strains 831:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 8 30 theme cell 1904:1907 arg1 integrity 1909:1917 cell integrity 1904:1917 cell integrity 1904:1917 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 7 31 theme OCH1 1422:1425 arg1 genes 1434:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 6 32 dep C. 1108:1109 arg1 neoformans 1111:1120 neoformans 1111:1120 neoformans 1111:1120 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 3 33 from residue 808:814 arg1 α1,6-linkage 774:785 an α1,6-linkage 771:785 an α1,6-linkage at the third mannose residue 771:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 5 34 theme strains 1011:1017 arg1 analyses 978:985 Further analyses 970:985 Further analyses of C. neoformans mutant strains 970:1017 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 7 35 theme Saccharomyces 1397:1409 arg1 genes 1434:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 6 36 theme second 1285:1290 arg1 residue 1300:1306 the second mannose residue 1281:1306 the second mannose residue 1281:1306 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 0 37 theme novel 15:19 arg1 structure 21:29 novel structure 15:29 novel structure 15:29 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 4 38 theme mutant 878:883 arg1 strains 885:891 wild-type and uxs1Δ mutant strains 858:891 wild-type and uxs1Δ mutant strains 858:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 3 39 theme short 681:685 arg1 manno-oligosaccharides 687:708 short manno-oligosaccharides 681:708 short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue 681:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 3 39 theme short 681:685 arg1 O-glycans 666:674 the major C. neoformans O-glycans 642:674 the major C. neoformans O-glycans 642:674 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 8 40 theme neoformans 1661:1670 arg1 strain 1685:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 3 41 theme detailed 491:498 arg1 analysis 511:518 a detailed structural analysis 489:518 a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins 489:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 8 42 theme mutant 1678:1683 arg1 strain 1685:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 8 43 theme high 1739:1742 arg1 salt 1744:1747 high salt 1739:1747 high salt 1739:1747 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 6 44 theme family 1187:1192 arg1 genes 1194:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 4 45 theme minor 927:931 arg1 O-glycans 933:941 minor O-glycans 927:941 minor O-glycans carrying a xylose residue 927:967 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 6 46 theme cerevisiae 1166:1175 arg1 genes 1194:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 8 47 theme high 1754:1757 arg1 temperature 1759:1769 high temperature 1754:1769 high temperature 1754:1769 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 3 48 theme exoglycosidase 601:614 arg1 treatment 616:624 exoglycosidase treatment 601:624 exoglycosidase treatment 601:624 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 7 49 contain containing 1573:1582 arg2 xylose 1584:1589 xylose 1584:1589 xylose 1584:1589 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 49 contain containing 1573:1582 arg1 O-glycans 1563:1571 minor O-glycans 1557:1571 minor O-glycans containing xylose 1557:1589 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 8 50 theme attenuated 1779:1788 arg1 virulence 1790:1798 attenuated virulence 1779:1798 attenuated virulence 1779:1798 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 3 51 theme cryptococcal 549:560 arg1 mannoproteins 562:574 cryptococcal mannoproteins 549:574 cryptococcal mannoproteins 549:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 7 52 theme mannose 1516:1522 arg1 residue 1524:1530 the third mannose residue 1506:1530 the third mannose residue 1506:1530 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 6 53 theme major 1335:1339 arg1 O-glycans 1341:1349 the major O-glycans 1331:1349 the major O-glycans 1331:1349 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 2 54 theme T-cell 347:352 arg1 response 354:361 the host T-cell response 338:361 the host T-cell response 338:361 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 5 55 from extensions 1083:1092 arg1 Golgi 1101:1105 the Golgi 1097:1105 the Golgi 1097:1105 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 1 56 theme encapsulated 184:195 arg1 basidiomycete 197:209 an encapsulated basidiomycete 181:209 an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans 181:260 Cryptococcus neoformans is an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans. 25477510 1 56 theme encapsulated 184:195 arg1 neoformans 167:176 Cryptococcus neoformans 154:176 Cryptococcus neoformans 154:176 Cryptococcus neoformans is an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans. 25477510 0 57 theme pathogenic 112:121 arg1 neoformans 142:151 the human pathogenic yeast Cryptococcus neoformans 102:151 the human pathogenic yeast Cryptococcus neoformans 102:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 3 58 theme mannose 800:806 arg1 residue 808:814 the third mannose residue 790:814 the third mannose residue 790:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 0 59 theme Cryptococcus 129:140 arg1 neoformans 142:151 the human pathogenic yeast Cryptococcus neoformans 102:151 the human pathogenic yeast Cryptococcus neoformans 102:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 2 60 theme cell 267:270 arg1 mannoproteins 280:292 The cell surface mannoproteins 263:292 The cell surface mannoproteins of C. neoformans 263:309 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 8 61 theme mouse 1805:1809 arg1 model 1811:1815 a mouse model 1803:1815 a mouse model of cryptococcosis 1803:1833 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 4 62 from strains 885:891 arg1 profiles 844:851 the O-glycan profiles 831:851 the O-glycan profiles from wild-type and uxs1Δ mutant strains 831:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 4 62 from strains 885:891 arg1 Comparison 817:826 Comparison 817:826 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains 817:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 4 63 theme xylose 954:959 arg1 residue 961:967 a xylose residue 952:967 a xylose residue 952:967 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 0 64 dep structure 21:29 arg1 the 11:13 the 11:13 the 11:13 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 3 65 theme neoformans 655:664 arg1 manno-oligosaccharides 687:708 short manno-oligosaccharides 681:708 short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue 681:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 3 65 theme neoformans 655:664 arg1 O-glycans 666:674 the major C. neoformans O-glycans 642:674 the major C. neoformans O-glycans 642:674 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 5 66 theme mannosyltransferase 1036:1054 arg1 genes 1056:1060 three mannosyltransferase genes 1030:1060 three mannosyltransferase genes involved in O-glycan extensions in the Golgi 1030:1105 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 0 67 theme glycans 68:74 arg1 structure 21:29 novel structure 15:29 novel structure 15:29 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 0 67 theme glycans 68:74 arg1 pathway 48:54 biosynthetic pathway 35:54 biosynthetic pathway 35:54 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 3 68 theme major 646:650 arg1 manno-oligosaccharides 687:708 short manno-oligosaccharides 681:708 short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue 681:814 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 3 68 theme major 646:650 arg1 O-glycans 666:674 the major C. neoformans O-glycans 642:674 the major C. neoformans O-glycans 642:674 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 0 69 from structure 21:29 arg1 apparatus 89:97 the Golgi apparatus 79:97 the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans 79:151 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 7 70 theme genes 1434:1438 arg1 homologs 1381:1388 homologs 1381:1388 homologs of the Saccharomyces cerevisiae OCH1 family genes 1381:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 70 theme genes 1434:1438 arg1 HOC1 1366:1369 HOC1 1366:1369 HOC1 1366:1369 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 8 71 theme extended 1856:1863 arg1 structure 1865:1873 the extended structure 1852:1873 the extended structure of O-glycans 1852:1886 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 1 72 theme Cryptococcus 154:165 arg1 basidiomycete 197:209 an encapsulated basidiomycete 181:209 an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans 181:260 Cryptococcus neoformans is an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans. 25477510 1 72 theme Cryptococcus 154:165 arg1 neoformans 167:176 Cryptococcus neoformans 154:176 Cryptococcus neoformans 154:176 Cryptococcus neoformans is an encapsulated basidiomycete causing cryptococcosis in immunocompromised humans. 25477510 7 73 dep C. 1352:1353 arg1 neoformans 1355:1364 neoformans 1355:1364 neoformans 1355:1364 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 4 74 from Comparison 817:826 arg1 strains 885:891 wild-type and uxs1Δ mutant strains 858:891 wild-type and uxs1Δ mutant strains 858:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 6 75 theme Saccharomyces 1152:1164 arg1 genes 1194:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 the Saccharomyces cerevisiae KRE2/MNT1 family genes 1148:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 5 76 theme mutant 1004:1009 arg1 strains 1011:1017 C. neoformans mutant strains 990:1017 C. neoformans mutant strains 990:1017 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 6 77 theme only 1132:1135 arg1 KTR3 1122:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 6 77 theme only 1132:1135 arg1 homolog 1137:1143 the only homolog 1128:1143 the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes 1128:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 0 78 theme biosynthetic 35:46 arg1 pathway 48:54 biosynthetic pathway 35:54 biosynthetic pathway 35:54 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 7 79 theme cerevisiae 1411:1420 arg1 genes 1434:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 the Saccharomyces cerevisiae OCH1 family genes 1393:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 8 80 theme C. 1658:1659 arg1 strain 1685:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 2 81 theme O-glycan 422:429 arg1 structure 431:439 their O-glycan structure 416:439 their O-glycan structure 416:439 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25477510 3 82 attach attached 537:544 arg2 O-glycans 527:535 the O-glycans 523:535 the O-glycans attached to cryptococcal mannoproteins 523:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 3 82 attach attached 537:544 arg1 mannoproteins 562:574 cryptococcal mannoproteins 549:574 cryptococcal mannoproteins 549:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 8 83 theme ktr3Δ 1672:1676 arg1 strain 1685:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 the C. neoformans ktr3Δ mutant strain 1654:1690 Moreover, the C. neoformans ktr3Δ mutant strain, which displayed increased sensitivity to SDS, high salt, and high temperature, showed attenuated virulence in a mouse model of cryptococcosis, suggesting that the extended structure of O-glycans is required for cell integrity and full pathogenicity of C. neoformans. 25477510 7 84 theme major 1598:1602 arg1 O-glycans 1604:1612 major O-glycans 1598:1612 major O-glycans 1598:1612 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 5 85 theme O-glycan 1074:1081 arg1 extensions 1083:1092 O-glycan extensions 1074:1092 O-glycan extensions in the Golgi 1074:1105 Further analyses of C. neoformans mutant strains identified three mannosyltransferase genes involved in O-glycan extensions in the Golgi. 25477510 0 86 link O-linked 59:66 arg1 glycans 68:74 O-linked glycans 59:74 O-linked glycans 59:74 Unraveling the novel structure and biosynthetic pathway of O-linked glycans in the Golgi apparatus of the human pathogenic yeast Cryptococcus neoformans. 25477510 7 87 theme C. 1352:1353 arg1 homologs 1381:1388 homologs 1381:1388 homologs of the Saccharomyces cerevisiae OCH1 family genes 1381:1438 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 87 theme C. 1352:1353 arg1 HOC3 1375:1378 HOC3 1375:1378 HOC3 1375:1378 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 87 theme C. 1352:1353 arg1 HOC1 1366:1369 HOC1 1366:1369 HOC1 1366:1369 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 7 87 theme C. 1352:1353 arg1 α1,6-mannosyltransferases 1462:1486 α1,6-mannosyltransferases 1462:1486 α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively 1462:1641 C. neoformans HOC1 and HOC3, homologs of the Saccharomyces cerevisiae OCH1 family genes, were shown to encode α1,6-mannosyltransferases that can transfer the third mannose residue, via an α1,6-linkage, to minor O-glycans containing xylose and to major O-glycans without xylose, respectively. 25477510 4 88 theme profiles 844:851 arg1 Comparison 817:826 Comparison 817:826 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains 817:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 3 89 theme structural 500:509 arg1 analysis 511:518 a detailed structural analysis 489:518 a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins 489:574 In this study, we performed a detailed structural analysis of the O-glycans attached to cryptococcal mannoproteins using HPLC combined with exoglycosidase treatment and showed that the major C. neoformans O-glycans were short manno-oligosaccharides that were connected mostly by α1,2-linkages but connected by an α1,6-linkage at the third mannose residue. 25477510 6 90 theme genes 1194:1198 arg1 KTR3 1122:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3 1108:1125 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 6 90 theme genes 1194:1198 arg1 homolog 1137:1143 the only homolog 1128:1143 the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes 1128:1198 C. neoformans KTR3, the only homolog of the Saccharomyces cerevisiae KRE2/MNT1 family genes, was shown to encode an α1,2-mannosyltransferase responsible for the addition of the second mannose residue via an α1,2-linkage to the major O-glycans. 25477510 4 91 theme wild-type 858:866 arg1 strains 885:891 wild-type and uxs1Δ mutant strains 858:891 wild-type and uxs1Δ mutant strains 858:891 Comparison of the O-glycan profiles from wild-type and uxs1Δ mutant strains strongly supports the presence of minor O-glycans carrying a xylose residue. 25477510 2 92 theme fungal 385:390 arg1 pathogenicity 392:404 fungal pathogenicity 385:404 fungal pathogenicity 385:404 The cell surface mannoproteins of C. neoformans were reported to stimulate the host T-cell response and to be involved in fungal pathogenicity; however, their O-glycan structure is uncharacterized. 25907685 1 0 theme biosimilars 254:264 arg1 " 265:265 "biosimilars" 253:265 "biosimilars" 253:265 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 12 1 theme Issue 2006:2010 arg1 part 1988:1991 part 1988:1991 part of a Special Issue entitled: HUPO 2014 1988:2030 This article is part of a Special Issue entitled: HUPO 2014. 25907685 10 2 from steps 1695:1699 arg1 process 1719:1725 the developing process 1704:1725 the developing process of biosimilars 1704:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 2 3 from targets 551:557 arg1 production 566:575 the production 562:575 the production of biosimilars 562:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 10 4 theme glycosylated 1755:1766 arg1 drug 1768:1771 glycosylated drug 1755:1771 any other glycosylated drug 1745:1771 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 3 5 with antibodies 625:634 arg1 pattern 681:687 the closest N-glycosylation pattern 653:687 the closest N-glycosylation pattern seen in the commercialized drug 653:719 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 3 5 with antibodies 625:634 arg1 same 645:648 same 645:648 same 645:648 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 10 6 theme N-glycosylation 1562:1576 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 6 theme N-glycosylation 1562:1576 arg1 sites 1578:1582 the N-glycosylation sites 1558:1582 the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1558:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 2 7 theme main 546:549 arg1 targets 551:557 the main targets 542:557 the main targets in the production of biosimilars 542:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 10 8 theme antibodies 1622:1631 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 8 theme antibodies 1622:1631 arg1 sites 1578:1582 the N-glycosylation sites 1558:1582 the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1558:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 9 theme recombinant 1599:1609 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 9 theme recombinant 1599:1609 arg1 mAbs 1634:1637 mAbs 1634:1637 mAbs 1634:1637 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 3 10 theme commercialized 701:714 arg1 drug 716:719 the commercialized drug 697:719 the commercialized drug 697:719 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 1 11 theme new 209:211 arg1 therapeutics 213:224 new therapeutics 209:224 new therapeutics 209:224 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 8 12 theme different 1261:1269 arg1 times 1271:1275 different times 1261:1275 different times 1261:1275 Different sugars will elute at different times and can be identified using specific oligosaccharide standards. 25907685 2 13 theme targets 551:557 arg1 one 535:537 one 535:537 one 535:537 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 13 theme targets 551:557 arg1 Trastuzumab 478:488 the ErbB2-targeted monoclonal antibody Trastuzumab 439:488 the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy 439:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 13 theme targets 551:557 arg1 targets 551:557 the main targets 542:557 the main targets in the production of biosimilars 542:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 3 14 theme major 595:599 arg1 challenge 601:609 A major challenge 593:609 A major challenge 593:609 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 9 15 theme sugar 1496:1500 arg1 masses 1502:1507 the differential sugar masses 1479:1507 the differential sugar masses 1479:1507 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 1 16 theme pharmaceutical 141:154 arg1 market 156:161 The pharmaceutical market 137:161 The pharmaceutical market 137:161 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 10 17 dep other 1749:1753 arg1 drug 1768:1771 glycosylated drug 1755:1771 any other glycosylated drug 1745:1771 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 7 18 theme Phase 1218:1222 arg1 HPLC 1224:1227 Normal Phase HPLC 1211:1227 Normal Phase HPLC 1211:1227 In the first case, N-glycans are fluorescently labeled and separated by Normal Phase HPLC. 25907685 6 19 gly N-glycosylation 1078:1092 arg1 candidates 1112:1121 two biosimilar candidates 1097:1121 two biosimilar candidates of Trastuzumab 1097:1136 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 8 20 theme oligosaccharide 1314:1328 arg1 standards 1330:1338 specific oligosaccharide standards 1305:1338 specific oligosaccharide standards 1305:1338 Different sugars will elute at different times and can be identified using specific oligosaccharide standards. 25907685 0 21 theme Liquid 86:91 arg1 Chromatography 93:106 Normal Phase Liquid Chromatography 73:106 Normal Phase Liquid Chromatography 73:106 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 2 22 theme breast 510:515 arg1 therapy 524:530 breast cancer therapy 510:530 breast cancer therapy 510:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 9 23 from approach 1355:1362 arg1 permethylated 1386:1398 permethylated 1386:1398 permethylated 1386:1398 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 0 24 theme Phase 80:84 arg1 Chromatography 93:106 Normal Phase Liquid Chromatography 73:106 Normal Phase Liquid Chromatography 73:106 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 11 25 theme labeling 1912:1919 arg1 use 1885:1887 the use 1881:1887 the use of glycan fluorescence labeling 1881:1919 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 0 26 theme N-glycosylation 0:14 arg1 analysis 24:31 N-glycosylation profile analysis 0:31 N-glycosylation profile analysis of Trastuzumab 0:46 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 4 27 theme Several 722:728 arg1 types 774:778 cell types 769:778 cell types 769:778 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 4 27 theme Several 722:728 arg1 conditions 755:764 growing conditions 747:764 growing conditions 747:764 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 4 27 theme Several 722:728 arg1 factors 730:736 Several factors 722:736 Several factors 722:736 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 10 28 theme consuming 1685:1693 arg1 steps 1695:1699 the most critical and time consuming steps 1658:1699 the most critical and time consuming steps in the developing process of biosimilars 1658:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 2 29 theme monoclonal 458:467 arg1 one 535:537 one 535:537 one 535:537 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 29 theme monoclonal 458:467 arg1 Herceptin 491:499 Herceptin 491:499 Herceptin 491:499 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 29 theme monoclonal 458:467 arg1 Trastuzumab 478:488 the ErbB2-targeted monoclonal antibody Trastuzumab 439:488 the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy 439:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 29 theme monoclonal 458:467 arg1 targets 551:557 the main targets 542:557 the main targets in the production of biosimilars 542:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 12 30 dep Issue 2006:2010 arg1 HUPO 2022:2025 HUPO 2014 2022:2030 a Special Issue entitled: HUPO 2014 1996:2030 This article is part of a Special Issue entitled: HUPO 2014. 25907685 0 31 theme Trastuzumab 36:46 arg1 analysis 24:31 N-glycosylation profile analysis 0:31 N-glycosylation profile analysis of Trastuzumab 0:46 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 9 32 theme second 1348:1353 arg1 approach 1355:1362 the second approach 1344:1362 the second approach 1344:1362 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 1 33 theme products 301:308 arg1 copies 268:273 copies 268:273 copies of already commercialized products waiting for the patents 268:332 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 0 34 theme Normal 73:78 arg1 Chromatography 93:106 Normal Phase Liquid Chromatography 73:106 Normal Phase Liquid Chromatography 73:106 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 10 35 theme developing 1708:1717 arg1 process 1719:1725 the developing process 1704:1725 the developing process of biosimilars 1704:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 11 36 dep HPLC 1932:1935 arg1 analysis 1962:1969 profile analysis 1954:1969 profile analysis 1954:1969 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 6 37 theme candidates 1112:1121 arg1 N-glycosylation 1078:1092 the N-glycosylation 1074:1092 the N-glycosylation of two biosimilar candidates of Trastuzumab 1074:1136 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 11 38 theme glycosylation 1857:1869 arg1 patterns 1871:1878 mAbs glycosylation patterns 1852:1878 mAbs glycosylation patterns 1852:1878 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 10 39 gly glycosylated 1755:1766 arg1 drug 1768:1771 glycosylated drug 1755:1771 any other glycosylated drug 1745:1771 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 11 40 theme MALDI-TOF 1941:1949 arg1 MS 1951:1952 MALDI-TOF MS 1941:1952 MALDI-TOF MS 1941:1952 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 4 41 theme cell 769:772 arg1 types 774:778 cell types 769:778 cell types 769:778 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 11 42 theme glycan 1892:1897 arg1 labeling 1912:1919 glycan fluorescence labeling 1892:1919 glycan fluorescence labeling 1892:1919 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 3 43 theme N-glycosylation 665:679 arg1 pattern 681:687 the closest N-glycosylation pattern 653:687 the closest N-glycosylation pattern seen in the commercialized drug 653:719 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 10 44 theme therapeutic 1587:1597 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 44 theme therapeutic 1587:1597 arg1 mAbs 1634:1637 mAbs 1634:1637 mAbs 1634:1637 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 9 45 theme MALDI-TOF 1416:1424 arg1 MS 1426:1427 MALDI-TOF MS 1416:1427 MALDI-TOF MS 1416:1427 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 12 46 theme Special 1998:2004 arg1 Issue 2006:2010 a Special Issue 1996:2010 a Special Issue entitled: HUPO 2014 1996:2030 This article is part of a Special Issue entitled: HUPO 2014. 25907685 10 47 theme sites 1578:1582 arg1 characterization 1538:1553 The characterization 1534:1553 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1510:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 2 48 theme biosimilars 580:590 arg1 production 566:575 the production 562:575 the production of biosimilars 562:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 8 49 theme Different 1230:1238 arg1 sugars 1240:1245 Different sugars 1230:1245 Different sugars 1230:1245 Different sugars will elute at different times and can be identified using specific oligosaccharide standards. 25907685 10 50 theme monoclonal 1611:1620 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 50 theme monoclonal 1611:1620 arg1 mAbs 1634:1637 mAbs 1634:1637 mAbs 1634:1637 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 11 51 theme complementary 1811:1823 arg1 approaches 1825:1834 two different but complementary approaches 1793:1834 two different but complementary approaches 1793:1834 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 6 52 theme Trastuzumab 1126:1136 arg1 candidates 1112:1121 two biosimilar candidates 1097:1121 two biosimilar candidates of Trastuzumab 1097:1136 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 11 53 theme different 1797:1805 arg1 approaches 1825:1834 two different but complementary approaches 1793:1834 two different but complementary approaches 1793:1834 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 1 54 theme therapeutics 213:224 arg1 production 195:204 the production 191:204 the production of new therapeutics 191:224 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 2 55 theme cancer 517:522 arg1 therapy 524:530 breast cancer therapy 510:530 breast cancer therapy 510:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 4 56 theme antibodies 912:921 arg1 properties 884:893 the properties 880:893 the properties of the generated antibodies 880:921 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 1 57 theme competitive 381:391 arg1 manners 408:414 a more competitive and affordable manners 374:414 a more competitive and affordable manners 374:414 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 10 58 theme biosimilars 1730:1740 arg1 process 1719:1725 the developing process 1704:1725 the developing process of biosimilars 1704:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 4 59 theme glycans 847:853 arg1 structure 830:838 structure 830:838 structure 830:838 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 4 59 theme glycans 847:853 arg1 composition 814:824 final composition 808:824 final composition 808:824 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 4 60 dep composition 814:824 arg1 the 804:806 the 804:806 the 804:806 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 10 61 gly N-glycosylation 1562:1576 arg1 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 61 gly N-glycosylation 1562:1576 arg2 sites 1578:1582 the N-glycosylation sites 1558:1582 the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1558:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 61 gly N-glycosylation 1562:1576 arg1 mAbs 1634:1637 mAbs 1634:1637 mAbs 1634:1637 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 61 gly N-glycosylation 1562:1576 arg2 antibodies 1622:1631 therapeutic recombinant monoclonal antibodies 1587:1631 therapeutic recombinant monoclonal antibodies (mAbs) 1587:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 1 62 theme affordable 397:406 arg1 manners 408:414 a more competitive and affordable manners 374:414 a more competitive and affordable manners 374:414 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 9 63 theme differential 1483:1494 arg1 masses 1502:1507 the differential sugar masses 1479:1507 the differential sugar masses 1479:1507 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 7 64 theme Normal 1211:1216 arg1 HPLC 1224:1227 Normal Phase HPLC 1211:1227 Normal Phase HPLC 1211:1227 In the first case, N-glycans are fluorescently labeled and separated by Normal Phase HPLC. 25907685 11 65 dep describe 1784:1791 arg1 coupled 1921:1927 coupled 1921:1927 coupled to HPLC and MALDI-TOF MS profile analysis 1921:1969 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 6 66 theme present 988:994 arg1 study 996:1000 the present study 984:1000 the present study 984:1000 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 10 67 dep SIGNIFICANCE 1521:1532 arg1 characterization 1538:1553 The characterization 1534:1553 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1510:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 2 68 theme antibody 469:476 arg1 one 535:537 one 535:537 one 535:537 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 68 theme antibody 469:476 arg1 Herceptin 491:499 Herceptin 491:499 Herceptin 491:499 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 68 theme antibody 469:476 arg1 Trastuzumab 478:488 the ErbB2-targeted monoclonal antibody Trastuzumab 439:488 the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy 439:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 68 theme antibody 469:476 arg1 targets 551:557 the main targets 542:557 the main targets in the production of biosimilars 542:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 8 69 theme specific 1305:1312 arg1 standards 1330:1338 specific oligosaccharide standards 1305:1338 specific oligosaccharide standards 1305:1338 Different sugars will elute at different times and can be identified using specific oligosaccharide standards. 25907685 0 70 theme MALDI-TOF 112:120 arg1 MS 122:123 MALDI-TOF MS 112:123 MALDI-TOF MS 112:123 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 4 71 theme generated 902:910 arg1 antibodies 912:921 the generated antibodies 898:921 the generated antibodies 898:921 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 5 72 theme appropriate 938:948 arg1 essential 970:978 essential 970:978 essential 970:978 Therefore, an appropriate characterization is essential. 25907685 5 72 theme appropriate 938:948 arg1 characterization 950:965 an appropriate characterization 935:965 an appropriate characterization 935:965 Therefore, an appropriate characterization is essential. 25907685 0 73 theme profile 16:22 arg1 analysis 24:31 N-glycosylation profile analysis 0:31 N-glycosylation profile analysis of Trastuzumab 0:46 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 0 74 dep Chromatography 93:106 arg1 approaches 125:134 approaches 125:134 approaches 125:134 N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches. 25907685 6 75 theme complementary 1033:1045 arg1 strategies 1047:1056 two different but complementary strategies 1015:1056 two different but complementary strategies 1015:1056 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 7 76 theme first 1146:1150 arg1 case 1152:1155 the first case 1142:1155 the first case 1142:1155 In the first case, N-glycans are fluorescently labeled and separated by Normal Phase HPLC. 25907685 6 77 theme different 1019:1027 arg1 strategies 1047:1056 two different but complementary strategies 1015:1056 two different but complementary strategies 1015:1056 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 11 78 theme profile 1954:1960 arg1 analysis 1962:1969 profile analysis 1954:1969 profile analysis 1954:1969 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 1 79 theme commercialized 286:299 arg1 products 301:308 already commercialized products 278:308 already commercialized products waiting for the patents 278:332 The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by "biosimilars", copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. 25907685 2 80 theme ErbB2-targeted 443:456 arg1 one 535:537 one 535:537 one 535:537 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 80 theme ErbB2-targeted 443:456 arg1 Herceptin 491:499 Herceptin 491:499 Herceptin 491:499 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 80 theme ErbB2-targeted 443:456 arg1 Trastuzumab 478:488 the ErbB2-targeted monoclonal antibody Trastuzumab 439:488 the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy 439:530 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 2 80 theme ErbB2-targeted 443:456 arg1 targets 551:557 the main targets 542:557 the main targets in the production of biosimilars 542:590 Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. 25907685 10 81 theme critical 1667:1674 arg1 steps 1695:1699 the most critical and time consuming steps 1658:1699 the most critical and time consuming steps in the developing process of biosimilars 1658:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 9 82 from permethylated 1386:1398 arg1 approach 1355:1362 the second approach 1344:1362 the second approach 1344:1362 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 4 83 theme final 808:812 arg1 composition 814:824 final composition 808:824 final composition 808:824 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 10 84 theme BIOLOGICAL 1510:1519 arg1 SIGNIFICANCE 1521:1532 BIOLOGICAL SIGNIFICANCE 1510:1532 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) 1510:1638 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 9 85 theme released 1365:1372 arg1 glycans 1374:1380 released glycans 1365:1380 released glycans 1365:1380 In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. 25907685 6 86 theme biosimilar 1101:1110 arg1 candidates 1112:1121 two biosimilar candidates 1097:1121 two biosimilar candidates of Trastuzumab 1097:1136 In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. 25907685 4 87 theme growing 747:753 arg1 conditions 755:764 growing conditions 747:764 growing conditions 747:764 Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. 25907685 10 88 theme steps 1695:1699 arg1 one 1651:1653 one 1651:1653 one 1651:1653 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 10 88 theme steps 1695:1699 arg1 steps 1695:1699 the most critical and time consuming steps 1658:1699 the most critical and time consuming steps in the developing process of biosimilars 1658:1740 BIOLOGICAL SIGNIFICANCE The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. 25907685 3 89 theme closest 657:663 arg1 pattern 681:687 the closest N-glycosylation pattern 653:687 the closest N-glycosylation pattern seen in the commercialized drug 653:719 A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. 25907685 11 90 theme fluorescence 1899:1910 arg1 labeling 1912:1919 glycan fluorescence labeling 1892:1919 glycan fluorescence labeling 1892:1919 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 25907685 11 91 theme mAbs 1852:1855 arg1 patterns 1871:1878 mAbs glycosylation patterns 1852:1878 mAbs glycosylation patterns 1852:1878 Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. 28785176 4 0 from forms 852:856 arg1 additions 824:832 O-fucose additions 815:832 O-fucose additions from their native forms 815:856 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 0 from forms 852:856 arg1 O-GlcNAc 800:807 O-GlcNAc 800:807 O-GlcNAc 800:807 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 0 from forms 852:856 arg1 O-GalNAc 766:773 O-GalNAc 766:773 O-GalNAc 766:773 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 0 from forms 852:856 arg1 O-N-acetylglucosamine 777:797 O-N-acetylglucosamine 777:797 O-N-acetylglucosamine (O-GlcNAc) 777:808 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 0 from forms 852:856 arg1 O-N-acetylgalactosamine 741:763 O-N-acetylgalactosamine 741:763 O-N-acetylgalactosamine (O-GalNAc) 741:774 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 1 theme native 845:850 arg1 forms 852:856 their native forms 839:856 their native forms 839:856 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 2 with peptides 1009:1016 arg1 modifications 1029:1041 these modifications 1023:1041 these modifications 1023:1041 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 2 3 theme O-glycosylated 300:313 arg1 peptides 315:322 O-glycosylated peptides 300:322 O-glycosylated peptides 300:322 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 4 4 used used 710:713 arg2 HILIC 701:705 HILIC 701:705 HILIC 701:705 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 2 5 gly O-glycosylated 300:313 arg1 peptides 315:322 O-glycosylated peptides 300:322 O-glycosylated peptides 300:322 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 1 6 theme O-Linked 70:77 arg1 glycosylation 79:91 O-Linked glycosylation 70:91 O-Linked glycosylation 70:91 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 6 theme O-Linked 70:77 arg1 modification 124:135 a common post-translational modification 96:135 a common post-translational modification that can alter the overall structure, polarity, and function of proteins 96:208 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 4 7 theme regression 944:953 arg1 analysis 955:962 linear regression analysis 937:962 linear regression analysis 937:962 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 7 theme regression 944:953 arg1 means 969:973 a means 967:973 a means to predict the retention times of peptides with these modifications 967:1041 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 8 theme O-fucose 815:822 arg1 additions 824:832 O-fucose additions 815:832 O-fucose additions from their native forms 815:856 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 3 9 theme interaction 462:472 arg1 solution 513:520 a solution 511:520 a solution to this problem 511:536 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 9 theme interaction 462:472 arg1 HILIC 497:501 HILIC 497:501 HILIC 497:501 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 9 theme interaction 462:472 arg1 chromatography 481:494 Hydrophilic interaction liquid chromatography 450:494 Hydrophilic interaction liquid chromatography (HILIC) 450:502 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 4 10 theme linear 937:942 arg1 analysis 955:962 linear regression analysis 937:962 linear regression analysis 937:962 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 10 theme linear 937:942 arg1 means 969:973 a means 967:973 a means to predict the retention times of peptides with these modifications 967:1041 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 3 11 from s 682:682 arg1 peptide 651:657 the peptide 647:657 the peptide from its modified form(s) 647:683 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 2 12 theme adequate 408:415 arg1 separation 417:426 adequate separation 408:426 adequate separation of these two species 408:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 13 theme unmodified 334:343 arg1 counterparts 345:356 their unmodified counterparts 328:356 their unmodified counterparts 328:356 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 1 14 theme common 98:103 arg1 glycosylation 79:91 O-Linked glycosylation 70:91 O-Linked glycosylation 70:91 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 14 theme common 98:103 arg1 modification 124:135 a common post-translational modification 96:135 a common post-translational modification that can alter the overall structure, polarity, and function of proteins 96:208 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 3 15 theme Hydrophilic 450:460 arg1 solution 513:520 a solution 511:520 a solution to this problem 511:536 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 15 theme Hydrophilic 450:460 arg1 HILIC 497:501 HILIC 497:501 HILIC 497:501 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 15 theme Hydrophilic 450:460 arg1 chromatography 481:494 Hydrophilic interaction liquid chromatography 450:494 Hydrophilic interaction liquid chromatography (HILIC) 450:502 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 1 16 theme post-translational 105:122 arg1 glycosylation 79:91 O-Linked glycosylation 70:91 O-Linked glycosylation 70:91 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 16 theme post-translational 105:122 arg1 modification 124:135 a common post-translational modification 96:135 a common post-translational modification that can alter the overall structure, polarity, and function of proteins 96:208 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 2 17 theme species 441:447 arg1 separation 417:426 adequate separation 408:426 adequate separation of these two species 408:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 3 18 theme polar 578:582 arg1 phase 595:599 the polar stationary phase 574:599 the polar stationary phase 574:599 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 0 19 theme Specific 37:44 arg1 Glycopeptides 55:67 Specific O-Linked Glycopeptides 37:67 Specific O-Linked Glycopeptides 37:67 Predicting the Retention Behavior of Specific O-Linked Glycopeptides. 28785176 4 20 with peptides 727:734 arg1 additions 824:832 O-fucose additions 815:832 O-fucose additions from their native forms 815:856 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 20 with peptides 727:734 arg1 O-GlcNAc 800:807 O-GlcNAc 800:807 O-GlcNAc 800:807 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 20 with peptides 727:734 arg1 O-GalNAc 766:773 O-GalNAc 766:773 O-GalNAc 766:773 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 20 with peptides 727:734 arg1 O-N-acetylglucosamine 777:797 O-N-acetylglucosamine 777:797 O-N-acetylglucosamine (O-GlcNAc) 777:808 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 20 with peptides 727:734 arg1 O-N-acetylgalactosamine 741:763 O-N-acetylgalactosamine 741:763 O-N-acetylgalactosamine (O-GalNAc) 741:774 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 4 21 theme hydrophilicity 903:916 arg1 extent 893:898 the extent 889:898 the extent of hydrophilicity 889:916 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 0 22 theme Glycopeptides 55:67 arg1 Behavior 25:32 the Retention Behavior 11:32 the Retention Behavior of Specific O-Linked Glycopeptides 11:67 Predicting the Retention Behavior of Specific O-Linked Glycopeptides. 28785176 4 23 theme peptides 1009:1016 arg1 times 1000:1004 the retention times 986:1004 the retention times of peptides with these modifications 986:1041 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 3 24 theme stationary 584:593 arg1 phase 595:599 the polar stationary phase 574:599 the polar stationary phase 574:599 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 0 25 theme O-Linked 46:53 arg1 Glycopeptides 55:67 Specific O-Linked Glycopeptides 37:67 Specific O-Linked Glycopeptides 37:67 Predicting the Retention Behavior of Specific O-Linked Glycopeptides. 28785176 3 26 theme form 677:680 arg1 s 682:682 its modified form(s) 664:683 its modified form(s) 664:683 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 2 27 theme RP 226:227 arg1 chromatography 230:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 27 theme RP 226:227 arg1 approach 280:287 the most common chromatographic approach 248:287 the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species 248:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 28 theme Reverse-phase 211:223 arg1 chromatography 230:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 28 theme Reverse-phase 211:223 arg1 approach 280:287 the most common chromatographic approach 248:287 the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species 248:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 29 theme chromatographic 264:278 arg1 chromatography 230:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 29 theme chromatographic 264:278 arg1 approach 280:287 the most common chromatographic approach 248:287 the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species 248:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 4 30 theme separate 718:725 arg1 peptides 727:734 separate peptides 718:734 separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms 718:856 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 2 31 theme common 257:262 arg1 chromatography 230:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography 211:243 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 2 31 theme common 257:262 arg1 approach 280:287 the most common chromatographic approach 248:287 the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species 248:447 Reverse-phase (RP) chromatography is the most common chromatographic approach to analyze O-glycosylated peptides and their unmodified counterparts, even though this approach often does not provide adequate separation of these two species. 28785176 3 32 theme modified 668:675 arg1 s 682:682 its modified form(s) 664:683 its modified form(s) 664:683 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 1 33 theme proteins 201:208 arg1 function 189:196 function 189:196 function 189:196 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 33 theme proteins 201:208 arg1 structure 164:172 the overall structure 152:172 the overall structure 152:172 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 33 theme proteins 201:208 arg1 polarity 175:182 polarity 175:182 polarity 175:182 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 1 34 theme overall 156:162 arg1 structure 164:172 the overall structure 152:172 the overall structure 152:172 O-Linked glycosylation is a common post-translational modification that can alter the overall structure, polarity, and function of proteins. 28785176 3 35 theme liquid 474:479 arg1 solution 513:520 a solution 511:520 a solution to this problem 511:536 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 35 theme liquid 474:479 arg1 HILIC 497:501 HILIC 497:501 HILIC 497:501 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 3 35 theme liquid 474:479 arg1 chromatography 481:494 Hydrophilic interaction liquid chromatography 450:494 Hydrophilic interaction liquid chromatography (HILIC) 450:502 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 4 36 theme retention 990:998 arg1 times 1000:1004 the retention times 986:1004 the retention times of peptides with these modifications 986:1041 In this paper, HILIC is used to separate peptides with O-N-acetylgalactosamine (O-GalNAc), O-N-acetylglucosamine (O-GlcNAc), and O-fucose additions from their native forms, and coefficients representing the extent of hydrophilicity were derived using linear regression analysis as a means to predict the retention times of peptides with these modifications. 28785176 3 37 theme polar 546:550 arg1 glycan 552:557 the polar glycan 542:557 the polar glycan 542:557 Hydrophilic interaction liquid chromatography (HILIC) can be a solution to this problem, as the polar glycan interacts with the polar stationary phase and potentially offers the ability to resolve the peptide from its modified form(s). 28785176 0 38 theme Retention 15:23 arg1 Behavior 25:32 the Retention Behavior 11:32 the Retention Behavior of Specific O-Linked Glycopeptides 11:67 Predicting the Retention Behavior of Specific O-Linked Glycopeptides. 27936757 9 0 theme GEP 1481:1483 arg1 data 1485:1488 publicly available GEP data 1462:1488 publicly available GEP data 1462:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 3 1 theme polyclonal 504:513 arg1 n 581:581 n = 5 581:585 n = 5 581:585 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 1 theme polyclonal 504:513 arg1 profiles 535:542 polyclonal IgG N-glycosylation profiles 504:542 polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8) 504:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 1 theme polyclonal 504:513 arg1 SMM 576:578 SMM 576:578 SMM 576:578 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 9 2 theme glycomic 1504:1511 arg1 changes 1513:1519 the glycomic changes 1500:1519 the glycomic changes seen in MM 1500:1530 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 6 3 theme afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2BG2S1 1076:1082 A2BG2S1 1076:1082 A2BG2S1 1076:1082 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 3 theme afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2BG2S2 1089:1095 A2BG2S2 1089:1095 A2BG2S2 1089:1095 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 3 theme afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2G2S1 1068:1073 A2G2S1 1068:1073 A2G2S1 1068:1073 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 3 theme afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 species 1059:1065 Three afucosyl-biantennary-digalactosylated-sialylated species 1004:1065 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) 1004:1096 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 4 gly afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2BG2S1 1076:1082 A2BG2S1 1076:1082 A2BG2S1 1076:1082 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 4 gly afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2BG2S2 1089:1095 A2BG2S2 1089:1095 A2BG2S2 1089:1095 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 4 gly afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 A2G2S1 1068:1073 A2G2S1 1068:1073 A2G2S1 1068:1073 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 4 gly afucosyl-biantennary-digalactosylated-sialylated 1010:1057 arg1 species 1059:1065 Three afucosyl-biantennary-digalactosylated-sialylated species 1004:1065 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) 1004:1096 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 10 5 theme myeloma 1629:1635 arg1 spectrum 1637:1644 the myeloma spectrum 1625:1644 the myeloma spectrum 1625:1644 The observed differential glycosylation underlined the heterogeneity of the myeloma spectrum. 27936757 7 6 theme transformation 1254:1267 arg1 case 1246:1249 a longitudinal case 1231:1249 a longitudinal case of transformation from MGUS to MM 1231:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 3 7 theme = 642:642 arg1 complete-response 601:617 complete-response (CR) post-treatment (n = 5) 601:645 complete-response (CR) post-treatment (n = 5) 601:645 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 7 theme = 642:642 arg1 n 640:640 n = 5 640:644 n = 5 640:644 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 2 8 theme immunoglobulin 283:296 arg1 G 298:298 immunoglobulin G 283:298 Abnormal immunoglobulin G (IgG) glycosylation in myeloma 274:329 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 2 8 theme immunoglobulin 283:296 arg1 IgG 301:303 IgG 301:303 IgG 301:303 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 3 9 dep n 581:581 arg1 MM 589:590 MM 589:590 MM (n = 8) 589:598 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 9 dep n 581:581 arg1 control 685:691 healthy age-matched control 665:691 healthy age-matched control (n = 5) 665:699 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 9 dep n 581:581 arg1 relapse 648:654 relapse 648:654 relapse (n = 4) 648:662 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 9 dep n 581:581 arg1 complete-response 601:617 complete-response (CR) post-treatment (n = 5) 601:645 complete-response (CR) post-treatment (n = 5) 601:645 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 9 dep n 581:581 arg1 n 593:593 n = 8 593:597 n = 8 593:597 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 6 10 theme active 1165:1170 arg1 myeloma 1172:1178 active myeloma 1165:1178 active myeloma 1165:1178 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 5 11 theme terminal 892:899 arg1 galactose 901:909 terminal galactose 892:909 terminal galactose 892:909 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 1 12 theme monoclonal 131:140 arg1 gammopathy 142:151 benign monoclonal gammopathy 124:151 benign monoclonal gammopathy of undetermined significance (MGUS) 124:187 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 1 12 theme monoclonal 131:140 arg1 MGUS 183:186 MGUS 183:186 MGUS 183:186 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 8 13 theme CR 1334:1335 arg1 group 1337:1341 the CR group 1330:1341 the CR group 1330:1341 Bisecting N-acetylglucosamine was lowest in the CR group, while highest in relapsed disease. 27936757 7 14 from MGUS 1274:1277 arg1 transformation 1254:1267 transformation 1254:1267 transformation from MGUS to MM 1254:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 7 14 from MGUS 1274:1277 arg1 case 1246:1249 a longitudinal case 1231:1249 a longitudinal case of transformation from MGUS to MM 1231:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 11 15 theme glycosylation 1793:1805 arg1 patterns 1807:1814 differential IgG glycosylation patterns 1776:1814 differential IgG glycosylation patterns 1776:1814 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 11 16 theme principle 1761:1769 arg1 proof 1752:1756 proof 1752:1756 proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders 1752:1870 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 0 17 theme Plasma 67:72 arg1 Disorders 79:87 Plasma Cell Disorders 67:87 Plasma Cell Disorders 67:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 11 18 theme plasma 1850:1855 arg1 disorders 1862:1870 plasma cell disorders 1850:1870 plasma cell disorders 1850:1870 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 9 19 theme Gene 1379:1382 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 4 20 theme normal 832:837 arg1 control 839:845 normal control 832:845 normal control 832:845 N-Glycan profiles from MGUS resembled normal control. 27936757 0 21 theme Disorders 79:87 arg1 Pathogenesis 51:62 the Pathogenesis 47:62 the Pathogenesis of Plasma Cell Disorders 47:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 8 22 theme relapsed 1361:1368 arg1 disease 1370:1376 relapsed disease 1361:1376 relapsed disease 1361:1376 Bisecting N-acetylglucosamine was lowest in the CR group, while highest in relapsed disease. 27936757 11 23 from molecule 1730:1737 arg1 feasibility 1675:1685 the feasibility 1671:1685 the feasibility of mapping glycan modifications on the IgG molecule 1671:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 3 24 from patients 552:559 arg1 n 581:581 n = 5 581:585 n = 5 581:585 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 24 from patients 552:559 arg1 profiles 535:542 polyclonal IgG N-glycosylation profiles 504:542 polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8) 504:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 24 from patients 552:559 arg1 SMM 576:578 SMM 576:578 SMM 576:578 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 25 theme age-matched 673:683 arg1 MM 589:590 MM 589:590 MM (n = 8) 589:598 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 25 theme age-matched 673:683 arg1 control 685:691 healthy age-matched control 665:691 healthy age-matched control (n = 5) 665:699 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 25 theme age-matched 673:683 arg1 n 694:694 n = 5 694:698 n = 5 694:698 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 5 26 theme agalactosylated 937:951 arg1 glycans 953:959 agalactosylated glycans 937:959 agalactosylated glycans 937:959 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 9 27 theme FUT 1405:1407 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 3 28 theme = 696:696 arg1 control 685:691 healthy age-matched control 665:691 healthy age-matched control (n = 5) 665:699 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 28 theme = 696:696 arg1 n 694:694 n = 5 694:698 n = 5 694:698 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 29 theme exoglycosidase 418:431 arg1 digestions 433:442 exoglycosidase digestions 418:442 exoglycosidase digestions 418:442 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 1 30 from gammopathy 142:151 arg1 progression 107:117 The pathological progression 90:117 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) 90:250 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 3 31 theme glycan 728:733 arg1 structures 735:744 glycan structures 728:744 glycan structures 728:744 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 7 32 theme chain 1197:1201 arg1 sialylation 1203:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 3 33 theme glycomic 358:365 arg1 platform 367:374 a glycomic platform 356:374 a glycomic platform composed of hydrophilic interaction UPLC 356:415 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 11 34 from modifications 1705:1717 arg1 molecule 1730:1737 the IgG molecule 1722:1737 the IgG molecule 1722:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 11 35 theme IgG 1789:1791 arg1 patterns 1807:1814 differential IgG glycosylation patterns 1776:1814 differential IgG glycosylation patterns 1776:1814 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 0 36 theme Polyclonal 0:9 arg1 N-Glycosylation 28:42 Polyclonal Immunoglobulin G N-Glycosylation 0:42 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders 0:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 1 37 theme pathological 94:105 arg1 progression 107:117 The pathological progression 90:117 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) 90:250 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 7 38 theme Increased 1181:1189 arg1 sialylation 1203:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 11 39 dep proof 1752:1756 arg1 identified 1836:1845 identified 1836:1845 can be successfully identified in plasma cell disorders 1816:1870 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 3 40 theme anion-exchange 450:463 arg1 chromatography 465:478 weak anion-exchange chromatography 445:478 weak anion-exchange chromatography 445:478 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 0 41 theme G 26:26 arg1 N-Glycosylation 28:42 Polyclonal Immunoglobulin G N-Glycosylation 0:42 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders 0:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 7 42 from case 1246:1249 arg1 MGUS 1274:1277 MGUS 1274:1277 MGUS 1274:1277 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 3 43 theme hydrophilic 388:398 arg1 UPLC 412:415 hydrophilic interaction UPLC 388:415 hydrophilic interaction UPLC 388:415 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 11 44 theme differential 1776:1787 arg1 patterns 1807:1814 differential IgG glycosylation patterns 1776:1814 differential IgG glycosylation patterns 1776:1814 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 5 45 theme neutral 865:871 arg1 glycans 873:879 neutral glycans 865:879 neutral glycans containing terminal galactose 865:909 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 3 46 theme mass 485:488 arg1 spectrometry 490:501 mass spectrometry 485:501 mass spectrometry 485:501 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 9 47 theme BACH2 1440:1444 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 3 48 theme myeloma 785:791 arg1 phases 766:771 distinct disease phases 749:771 distinct disease phases of multiple myeloma 749:791 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 5 49 contain containing 881:890 arg1 glycans 873:879 neutral glycans 865:879 neutral glycans containing terminal galactose 865:909 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 5 49 contain containing 881:890 arg2 galactose 901:909 terminal galactose 892:909 terminal galactose 892:909 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 11 50 theme cell 1857:1860 arg1 disorders 1862:1870 plasma cell disorders 1850:1870 plasma cell disorders 1850:1870 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 11 51 theme glycan 1698:1703 arg1 modifications 1705:1717 mapping glycan modifications 1690:1717 mapping glycan modifications on the IgG molecule 1690:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 1 52 theme significance 169:180 arg1 gammopathy 142:151 benign monoclonal gammopathy 124:151 benign monoclonal gammopathy of undetermined significance (MGUS) 124:187 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 1 52 theme significance 169:180 arg1 MGUS 183:186 MGUS 183:186 MGUS 183:186 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 3 53 theme = 595:595 arg1 MM 589:590 MM 589:590 MM (n = 8) 589:598 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 53 theme = 595:595 arg1 n 593:593 n = 8 593:597 n = 8 593:597 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 54 theme IgG 515:517 arg1 n 581:581 n = 5 581:585 n = 5 581:585 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 54 theme IgG 515:517 arg1 profiles 535:542 polyclonal IgG N-glycosylation profiles 504:542 polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8) 504:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 54 theme IgG 515:517 arg1 SMM 576:578 SMM 576:578 SMM 576:578 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 4 55 from MGUS 817:820 arg1 profiles 803:810 N-Glycan profiles 794:810 N-Glycan profiles from MGUS 794:820 N-Glycan profiles from MGUS resembled normal control. 27936757 9 56 theme available 1471:1479 arg1 data 1485:1488 publicly available GEP data 1462:1488 publicly available GEP data 1462:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 11 57 theme IgG 1726:1728 arg1 molecule 1730:1737 the IgG molecule 1722:1737 the IgG molecule 1722:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 11 58 from feasibility 1675:1685 arg1 molecule 1730:1737 the IgG molecule 1722:1737 the IgG molecule 1722:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 2 59 from glycosylation 306:318 arg1 myeloma 323:329 myeloma 323:329 myeloma 323:329 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 2 60 theme Abnormal 274:281 arg1 glycosylation 306:318 Abnormal immunoglobulin G (IgG) glycosylation 274:318 Abnormal immunoglobulin G (IgG) glycosylation in myeloma 274:329 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 3 61 theme post-treatment 624:637 arg1 MM 589:590 MM 589:590 MM (n = 8) 589:598 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 61 theme post-treatment 624:637 arg1 CR 620:621 CR 620:621 CR 620:621 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 61 theme post-treatment 624:637 arg1 complete-response 601:617 complete-response (CR) post-treatment (n = 5) 601:645 complete-response (CR) post-treatment (n = 5) 601:645 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 61 theme post-treatment 624:637 arg1 n 640:640 n = 5 640:644 n = 5 640:644 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 10 62 theme spectrum 1637:1644 arg1 heterogeneity 1608:1620 the heterogeneity 1604:1620 the heterogeneity of the myeloma spectrum 1604:1644 The observed differential glycosylation underlined the heterogeneity of the myeloma spectrum. 27936757 1 63 theme smoldering 192:201 arg1 SMM 212:214 SMM 212:214 SMM 212:214 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 1 63 theme smoldering 192:201 arg1 myeloma 203:209 smoldering myeloma 192:209 smoldering myeloma (SMM) 192:215 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 2 64 theme G 298:298 arg1 glycosylation 306:318 Abnormal immunoglobulin G (IgG) glycosylation 274:318 Abnormal immunoglobulin G (IgG) glycosylation in myeloma 274:329 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 3 65 theme = 570:570 arg1 patients 552:559 35 patients [MGUS 549:565 35 patients [MGUS (n = 8) 549:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 65 theme = 570:570 arg1 n 568:568 n = 8 568:572 n = 8 568:572 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 5 66 gly fucosylated 965:975 arg1 glycans 977:983 fucosylated glycans 965:983 fucosylated glycans 965:983 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 6 67 dep species 1059:1065 arg1 A2BG2S1 1076:1082 A2BG2S1 1076:1082 A2BG2S1 1076:1082 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 67 dep species 1059:1065 arg1 A2BG2S2 1089:1095 A2BG2S2 1089:1095 A2BG2S2 1089:1095 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 67 dep species 1059:1065 arg1 A2G2S1 1068:1073 A2G2S1 1068:1073 A2G2S1 1068:1073 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 6 67 dep species 1059:1065 arg1 species 1059:1065 Three afucosyl-biantennary-digalactosylated-sialylated species 1004:1065 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) 1004:1096 Three afucosyl-biantennary-digalactosylated-sialylated species (A2G2S1, A2BG2S1, and A2BG2S2) decreased 2.38-, 2.4-, and 4.25-fold, respectively, from benign to active myeloma. 27936757 3 68 theme = 659:659 arg1 n 657:657 n = 4 657:661 n = 4 657:661 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 68 theme = 659:659 arg1 relapse 648:654 relapse 648:654 relapse (n = 4) 648:662 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 1 69 theme benign 124:129 arg1 gammopathy 142:151 benign monoclonal gammopathy 124:151 benign monoclonal gammopathy of undetermined significance (MGUS) 124:187 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 1 69 theme benign 124:129 arg1 MGUS 183:186 MGUS 183:186 MGUS 183:186 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 7 70 theme longitudinal 1233:1244 arg1 case 1246:1249 a longitudinal case 1231:1249 a longitudinal case of transformation from MGUS to MM 1231:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 3 71 theme = 583:583 arg1 n 581:581 n = 5 581:585 n = 5 581:585 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 71 theme = 583:583 arg1 profiles 535:542 polyclonal IgG N-glycosylation profiles 504:542 polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8) 504:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 72 theme [MGUS 561:565 arg1 patients 552:559 35 patients [MGUS 549:565 35 patients [MGUS (n = 8) 549:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 72 theme [MGUS 561:565 arg1 n 568:568 n = 8 568:572 n = 8 568:572 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 73 theme healthy 665:671 arg1 MM 589:590 MM 589:590 MM (n = 8) 589:598 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 73 theme healthy 665:671 arg1 control 685:691 healthy age-matched control 665:691 healthy age-matched control (n = 5) 665:699 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 73 theme healthy 665:671 arg1 n 694:694 n = 5 694:698 n = 5 694:698 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 0 74 theme Cell 74:77 arg1 Disorders 79:87 Plasma Cell Disorders 67:87 Plasma Cell Disorders 67:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 1 75 theme undetermined 156:167 arg1 significance 169:180 undetermined significance 156:180 undetermined significance 156:180 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 0 76 from N-Glycosylation 28:42 arg1 Pathogenesis 51:62 the Pathogenesis 47:62 the Pathogenesis of Plasma Cell Disorders 47:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 7 77 gly sialylation 1203:1213 arg1 case 1246:1249 a longitudinal case 1231:1249 a longitudinal case of transformation from MGUS to MM 1231:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 9 78 theme expression 1384:1393 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 11 79 theme modifications 1705:1717 arg1 feasibility 1675:1685 the feasibility 1671:1685 the feasibility of mapping glycan modifications on the IgG molecule 1671:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 27936757 3 80 theme distinct 749:756 arg1 phases 766:771 distinct disease phases 749:771 distinct disease phases of multiple myeloma 749:791 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 7 81 theme light 1191:1195 arg1 sialylation 1203:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 4 82 theme N-Glycan 794:801 arg1 profiles 803:810 N-Glycan profiles 794:810 N-Glycan profiles from MGUS 794:820 N-Glycan profiles from MGUS resembled normal control. 27936757 0 83 theme Immunoglobulin 11:24 arg1 N-Glycosylation 28:42 Polyclonal Immunoglobulin G N-Glycosylation 0:42 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders 0:87 Polyclonal Immunoglobulin G N-Glycosylation in the Pathogenesis of Plasma Cell Disorders. 27936757 5 84 theme fucosylated 965:975 arg1 glycans 977:983 fucosylated glycans 965:983 fucosylated glycans 965:983 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 9 85 theme ST6GAL1 1412:1418 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 9 86 theme B4GALT1 1421:1427 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 3 87 theme multiple 776:783 arg1 myeloma 785:791 multiple myeloma 776:791 multiple myeloma 776:791 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 10 88 theme differential 1566:1577 arg1 glycosylation 1579:1591 The observed differential glycosylation 1553:1591 The observed differential glycosylation 1553:1591 The observed differential glycosylation underlined the heterogeneity of the myeloma spectrum. 27936757 9 89 theme RECK 1430:1433 arg1 levels 1395:1400 Gene expression levels 1379:1400 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data 1379:1488 Gene expression levels of FUT 8, ST6GAL1, B4GALT1, RECK, and BACH2 identified from publicly available GEP data supported the glycomic changes seen in MM compared to control. 27936757 3 90 theme interaction 400:410 arg1 UPLC 412:415 hydrophilic interaction UPLC 388:415 hydrophilic interaction UPLC 388:415 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 1 91 theme active 232:237 arg1 MM 248:249 MM 248:249 MM 248:249 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 1 91 theme active 232:237 arg1 myeloma 239:245 active myeloma 232:245 active myeloma (MM) 232:250 The pathological progression from benign monoclonal gammopathy of undetermined significance (MGUS) to smoldering myeloma (SMM) and finally to active myeloma (MM) is poorly understood. 27936757 3 92 theme weak 445:448 arg1 chromatography 465:478 weak anion-exchange chromatography 445:478 weak anion-exchange chromatography 445:478 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 93 theme disease 758:764 arg1 phases 766:771 distinct disease phases 749:771 distinct disease phases of multiple myeloma 749:791 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 10 94 gly heterogeneity 1608:1620 arg1 spectrum 1637:1644 the myeloma spectrum 1625:1644 the myeloma spectrum 1625:1644 The observed differential glycosylation underlined the heterogeneity of the myeloma spectrum. 27936757 5 95 theme glycans 873:879 arg1 abundance 852:860 The abundance 848:860 The abundance of neutral glycans containing terminal galactose 848:909 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 5 95 theme glycans 873:879 arg1 highest 915:921 highest 915:921 highest 915:921 The abundance of neutral glycans containing terminal galactose was highest in SMM, while agalactosylated glycans and fucosylated glycans were lowest in MM. 27936757 2 96 gly glycosylation 306:318 arg1 myeloma 323:329 myeloma 323:329 myeloma 323:329 Abnormal immunoglobulin G (IgG) glycosylation in myeloma has been reported. 27936757 10 97 theme observed 1557:1564 arg1 glycosylation 1579:1591 The observed differential glycosylation 1553:1591 The observed differential glycosylation 1553:1591 The observed differential glycosylation underlined the heterogeneity of the myeloma spectrum. 27936757 7 98 located observed 1219:1226 arg2 sialylation 1203:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation 1181:1213 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 7 98 located observed 1219:1226 arg1 case 1246:1249 a longitudinal case 1231:1249 a longitudinal case of transformation from MGUS to MM 1231:1283 Increased light chain sialylation was observed in a longitudinal case of transformation from MGUS to MM. 27936757 8 99 theme Bisecting 1286:1294 arg1 N-acetylglucosamine 1296:1314 Bisecting N-acetylglucosamine 1286:1314 Bisecting N-acetylglucosamine 1286:1314 Bisecting N-acetylglucosamine was lowest in the CR group, while highest in relapsed disease. 27936757 3 100 theme N-glycosylation 519:533 arg1 n 581:581 n = 5 581:585 n = 5 581:585 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 100 theme N-glycosylation 519:533 arg1 profiles 535:542 polyclonal IgG N-glycosylation profiles 504:542 polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8) 504:573 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 3 100 theme N-glycosylation 519:533 arg1 SMM 576:578 SMM 576:578 SMM 576:578 Using a glycomic platform composed of hydrophilic interaction UPLC, exoglycosidase digestions, weak anion-exchange chromatography, and mass spectrometry, polyclonal IgG N-glycosylation profiles from 35 patients [MGUS (n = 8), SMM (n = 5), MM (n = 8), complete-response (CR) post-treatment (n = 5), relapse (n = 4), healthy age-matched control (n = 5)] were characterized to map glycan structures in distinct disease phases of multiple myeloma. 27936757 11 101 theme mapping 1690:1696 arg1 modifications 1705:1717 mapping glycan modifications 1690:1717 mapping glycan modifications on the IgG molecule 1690:1737 This study demonstrates the feasibility of mapping glycan modifications on the IgG molecule and provides proof of principle that differential IgG glycosylation patterns can be successfully identified in plasma cell disorders. 28444691 1 0 gly glycosylation 142:154 arg1 glycoconjugates 159:173 glycoconjugates 159:173 glycoconjugates 159:173 Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. 28444691 7 1 theme triantennary 1250:1261 arg1 glycans 1263:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 6 2 theme significant 977:987 arg1 increase 989:996 a significant increase 975:996 a significant increase of apoC-III0a (aglycosylated glycoform) 975:1036 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 7 3 theme biantennary 1234:1244 arg1 glycans 1263:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 2 4 theme subunits 268:275 arg1 defects 241:247 defects 241:247 defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 241:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 4 5 theme acquired 684:691 arg1 microcephaly 693:704 acquired microcephaly 684:704 acquired microcephaly 684:704 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 2 6 theme COG 256:258 arg1 subunits 268:275 the COG complex subunits 1-8 252:279 the COG complex subunits 1-8 252:279 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 8 7 from knowledge 1539:1547 arg1 defects 1566:1572 glycosylation defects 1552:1572 glycosylation defects caused by impairment of COG subunits 1552:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 4 8 theme degree 666:671 arg1 features 620:627 dysmorphic features 609:627 dysmorphic features 609:627 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 8 theme degree 666:671 arg1 seizures 674:681 seizures 674:681 seizures 674:681 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 8 theme degree 666:671 arg1 defects 715:721 sensory defects 707:721 sensory defects 707:721 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 8 theme degree 666:671 arg1 disability 643:652 intellectual disability 630:652 intellectual disability of variable degree 630:671 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 8 theme degree 666:671 arg1 microcephaly 693:704 acquired microcephaly 684:704 acquired microcephaly 684:704 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 8 theme degree 666:671 arg1 behavior 736:743 autistic behavior 727:743 autistic behavior 727:743 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 7 9 theme relative 1172:1179 arg1 abundance 1181:1189 the relative abundance 1168:1189 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 10 theme slight 1282:1287 arg1 increase 1289:1296 slight increase 1282:1296 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 11 theme Serum 1110:1114 arg1 N-glycome 1116:1124 Serum N-glycome 1110:1124 Serum N-glycome of COG5-CDG patients 1110:1145 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 8 12 theme subunits 1602:1609 arg1 impairment 1584:1593 impairment 1584:1593 impairment of COG subunits 1584:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 5 13 from screening 826:834 arg1 patients 894:901 five patients 889:901 five patients with COG5-CDG 889:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 7 14 theme COG5-CDG 1129:1136 arg1 patients 1138:1145 COG5-CDG patients 1129:1145 COG5-CDG patients 1129:1145 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 8 15 theme N-glycan 1460:1467 arg1 profiling 1469:1477 N-glycan profiling 1460:1477 N-glycan profiling 1460:1477 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 5 16 theme high-throughput 810:824 arg1 screening 826:834 a high-throughput screening 808:834 a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 808:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 5 17 from O-glycoform 858:868 arg1 patients 894:901 five patients 889:901 five patients with COG5-CDG 889:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 4 18 with disease 596:602 arg1 features 620:627 dysmorphic features 609:627 dysmorphic features 609:627 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 18 with disease 596:602 arg1 seizures 674:681 seizures 674:681 seizures 674:681 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 18 with disease 596:602 arg1 defects 715:721 sensory defects 707:721 sensory defects 707:721 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 18 with disease 596:602 arg1 disability 643:652 intellectual disability 630:652 intellectual disability of variable degree 630:671 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 18 with disease 596:602 arg1 microcephaly 693:704 acquired microcephaly 684:704 acquired microcephaly 684:704 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 18 with disease 596:602 arg1 behavior 736:743 autistic behavior 727:743 autistic behavior 727:743 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 8 19 theme diagnosis 1635:1643 arg1 support 1615:1621 support 1615:1621 support of clinical diagnosis 1615:1643 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 4 20 theme autistic 727:734 arg1 behavior 736:743 autistic behavior 727:743 autistic behavior 727:743 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 8 21 from O-glycan 1447:1454 arg1 patients 1491:1498 COG5-CDG patients 1482:1498 COG5-CDG patients 1482:1498 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 1 22 theme glycoconjugates 159:173 arg1 glycosylation 142:154 defective glycosylation 132:154 defective glycosylation of glycoconjugates 132:173 Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. 28444691 7 23 theme glycans 1336:1342 arg1 increase 1289:1296 slight increase 1282:1296 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 23 theme glycans 1336:1342 arg1 abundance 1181:1189 the relative abundance 1168:1189 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 8 24 from aspects 1436:1442 arg1 O-glycan 1447:1454 O-glycan 1447:1454 O-glycan 1447:1454 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 8 24 from aspects 1436:1442 arg1 profiling 1469:1477 N-glycan profiling 1460:1477 N-glycan profiling 1460:1477 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 6 25 theme mono-sialylated 1058:1072 arg1 apoC-III1 1047:1055 apoC-III1 1047:1055 apoC-III1 (mono-sialylated glycoform) 1047:1083 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 6 25 theme mono-sialylated 1058:1072 arg1 glycoform 1074:1082 mono-sialylated glycoform 1058:1082 mono-sialylated glycoform 1058:1082 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 5 26 from N-glycoform 874:884 arg1 patients 894:901 five patients 889:901 five patients with COG5-CDG 889:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 3 27 gly undersialylated 384:398 arg1 apoC-III 458:465 apoC-III 458:465 apoC-III 458:465 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 27 gly undersialylated 384:398 arg1 glycoforms 400:409 undersialylated glycoforms 384:409 undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 384:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 27 gly undersialylated 384:398 arg1 transferrin 420:430 serum transferrin 414:430 serum transferrin 414:430 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 27 gly undersialylated 384:398 arg1 C-III 451:455 apolipoprotein C-III 436:455 apolipoprotein C-III (apoC-III) 436:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 28 theme undersialylated 384:398 arg1 glycoforms 400:409 undersialylated glycoforms 384:409 undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 384:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 29 theme apolipoprotein 436:449 arg1 apoC-III 458:465 apoC-III 458:465 apoC-III 458:465 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 29 theme apolipoprotein 436:449 arg1 C-III 451:455 apolipoprotein C-III 436:455 apolipoprotein C-III (apoC-III) 436:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 0 30 theme MALDI-MS 0:7 arg1 profiling 9:17 MALDI-MS profiling 0:17 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG 0:74 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. 28444691 1 31 theme glycosylation 101:113 arg1 disorders 88:96 Congenital disorders 77:96 Congenital disorders of glycosylation (CDG) 77:119 Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. 28444691 3 32 theme transferrin 420:430 arg1 glycoforms 400:409 undersialylated glycoforms 384:409 undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 384:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 2 33 theme oligomeric 186:195 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 33 theme oligomeric 186:195 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 8 34 theme knowledge 1539:1547 arg1 increase 1519:1526 an increase 1516:1526 an increase of current knowledge on glycosylation defects caused by impairment of COG subunits 1516:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 0 35 theme O-glycosylation 28:42 arg1 profiling 9:17 MALDI-MS profiling 0:17 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG 0:74 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. 28444691 5 36 theme serum 852:856 arg1 O-glycoform 858:868 differential serum O-glycoform 839:868 differential serum O-glycoform 839:868 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 8 37 theme glycosylation 1552:1564 arg1 defects 1566:1572 glycosylation defects 1552:1572 glycosylation defects caused by impairment of COG subunits 1552:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 3 38 theme O-glycosylation 505:519 arg1 defects 521:527 N-glycosylation and O-glycosylation defects 485:527 N-glycosylation and O-glycosylation defects 485:527 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 0 39 theme N-glycosylation 48:62 arg1 profiling 9:17 MALDI-MS profiling 0:17 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG 0:74 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. 28444691 4 40 theme subunit 559:565 arg1 deficiency 567:576 COG5 subunit deficiency 554:576 COG5 subunit deficiency 554:576 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 40 theme subunit 559:565 arg1 COG5-CDG 544:551 COG5-CDG 544:551 COG5-CDG (COG5 subunit deficiency) 544:577 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 8 41 theme current 1531:1537 arg1 knowledge 1539:1547 current knowledge 1531:1547 current knowledge on glycosylation defects caused by impairment of COG subunits 1531:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 3 42 theme N-glycosylation 485:499 arg1 defects 521:527 N-glycosylation and O-glycosylation defects 485:527 N-glycosylation and O-glycosylation defects 485:527 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 8 43 theme well-established 1364:1379 arg1 approaches 1390:1399 advanced and well-established MS-based approaches 1351:1399 advanced and well-established MS-based approaches 1351:1399 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 5 44 theme O-glycoform 858:868 arg1 screening 826:834 a high-throughput screening 808:834 a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 808:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 6 45 theme apoC-III0a 1001:1010 arg1 increase 989:996 a significant increase 975:996 a significant increase of apoC-III0a (aglycosylated glycoform) 975:1036 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 8 46 from increase 1519:1526 arg1 defects 1566:1572 glycosylation defects 1552:1572 glycosylation defects caused by impairment of COG subunits 1552:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 7 47 theme undergalactosylated 1214:1232 arg1 glycans 1263:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 4 48 theme dysmorphic 609:618 arg1 features 620:627 dysmorphic features 609:627 dysmorphic features 609:627 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 7 49 theme undersialylated 1194:1208 arg1 glycans 1263:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 6 50 gly aglycosylated 1013:1025 arg1 glycoform 1027:1035 aglycosylated glycoform 1013:1035 aglycosylated glycoform 1013:1035 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 6 50 gly aglycosylated 1013:1025 arg1 apoC-III0a 1001:1010 apoC-III0a 1001:1010 apoC-III0a (aglycosylated glycoform) 1001:1036 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 2 51 theme complex 260:266 arg1 subunits 268:275 the COG complex subunits 1-8 252:279 the COG complex subunits 1-8 252:279 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 7 52 gly undersialylated 1194:1208 arg1 glycans 1263:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 undersialylated and undergalactosylated biantennary and triantennary glycans 1194:1269 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 2 53 theme O-glycan 302:309 arg1 abnormalities 322:334 O-glycan processing abnormalities 302:334 O-glycan processing abnormalities 302:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 5 54 with patients 894:901 arg1 COG5-CDG 908:915 COG5-CDG 908:915 COG5-CDG 908:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 8 55 theme COG 1598:1600 arg1 subunits 1602:1609 COG subunits 1598:1609 COG subunits 1598:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 4 56 theme variable 657:664 arg1 degree 666:671 variable degree 657:671 variable degree 657:671 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 2 57 theme due 234:236 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 57 theme due 234:236 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 8 58 from defects 1566:1572 arg1 increase 1519:1526 an increase 1516:1526 an increase of current knowledge on glycosylation defects caused by impairment of COG subunits 1516:1609 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 7 59 theme hybrid 1329:1334 arg1 glycans 1336:1342 hybrid glycans 1329:1342 hybrid glycans 1329:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 60 theme patients 1138:1145 arg1 N-glycome 1116:1124 Serum N-glycome 1110:1124 Serum N-glycome of COG5-CDG patients 1110:1145 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 2 61 theme genetic 217:223 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 61 theme genetic 217:223 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 8 62 theme advanced 1351:1358 arg1 approaches 1390:1399 advanced and well-established MS-based approaches 1351:1399 advanced and well-established MS-based approaches 1351:1399 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 6 63 theme aglycosylated 1013:1025 arg1 glycoform 1027:1035 aglycosylated glycoform 1013:1035 aglycosylated glycoform 1013:1035 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 6 63 theme aglycosylated 1013:1025 arg1 apoC-III0a 1001:1010 apoC-III0a 1001:1010 apoC-III0a (aglycosylated glycoform) 1001:1036 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 8 64 theme novel 1430:1434 arg1 aspects 1436:1442 novel aspects 1430:1442 novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients 1430:1498 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 8 65 theme clinical 1626:1633 arg1 diagnosis 1635:1643 clinical diagnosis 1626:1643 clinical diagnosis 1626:1643 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 8 66 theme present 1406:1412 arg1 findings 1414:1421 the present findings 1402:1421 the present findings 1402:1421 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 1 67 theme defective 132:140 arg1 glycosylation 142:154 defective glycosylation 132:154 defective glycosylation of glycoconjugates 132:173 Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. 28444691 4 68 theme sensory 707:713 arg1 defects 715:721 sensory defects 707:721 sensory defects 707:721 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 5 69 theme laser 773:777 arg1 desorption/ionization-MS 779:802 matrix-assisted laser desorption/ionization-MS 757:802 matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 757:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 0 70 from profiling 9:17 arg1 COG5-CDG 67:74 COG5-CDG 67:74 COG5-CDG 67:74 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. 28444691 2 71 dep subunits 268:275 arg1 1-8 277:279 1-8 277:279 1-8 277:279 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 5 72 theme matrix-assisted 757:771 arg1 desorption/ionization-MS 779:802 matrix-assisted laser desorption/ionization-MS 757:802 matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 757:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 3 73 theme focusing 361:368 arg1 separation 370:379 isoelectric focusing separation 349:379 isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 349:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 8 74 theme COG5-CDG 1482:1489 arg1 patients 1491:1498 COG5-CDG patients 1482:1498 COG5-CDG patients 1482:1498 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 7 75 theme structures 1314:1323 arg1 increase 1289:1296 slight increase 1282:1296 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 75 theme structures 1314:1323 arg1 abundance 1181:1189 the relative abundance 1168:1189 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 5 76 from patients 894:901 arg1 screening 826:834 a high-throughput screening 808:834 a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 808:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 3 77 gly glycoforms 400:409 arg1 apoC-III 458:465 apoC-III 458:465 apoC-III 458:465 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 77 gly glycoforms 400:409 arg1 transferrin 420:430 serum transferrin 414:430 serum transferrin 414:430 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 3 77 gly glycoforms 400:409 arg1 C-III 451:455 apolipoprotein C-III 436:455 apolipoprotein C-III (apoC-III) 436:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 1 78 theme Congenital 77:86 arg1 disorders 88:96 Congenital disorders 77:96 Congenital disorders of glycosylation (CDG) 77:119 Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. 28444691 5 79 theme N-glycoform 874:884 arg1 screening 826:834 a high-throughput screening 808:834 a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG 808:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 8 80 from profiling 1469:1477 arg1 patients 1491:1498 COG5-CDG patients 1482:1498 COG5-CDG patients 1482:1498 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 3 81 theme glycoforms 400:409 arg1 separation 370:379 isoelectric focusing separation 349:379 isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 349:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 8 82 theme MS-based 1381:1388 arg1 approaches 1390:1399 advanced and well-established MS-based approaches 1351:1399 advanced and well-established MS-based approaches 1351:1399 Using advanced and well-established MS-based approaches, the present findings reveal novel aspects on O-glycan and N-glycan profiling in COG5-CDG patients, thus providing an increase of current knowledge on glycosylation defects caused by impairment of COG subunits, in support of clinical diagnosis. 28444691 9 83 dep & 1674:1674 arg1 Sons 1676:1679 Sons 1676:1679 Sons 1676:1679 Copyright © 2017 John Wiley & Sons, Ltd. 28444691 3 84 theme C-III 451:455 arg1 glycoforms 400:409 undersialylated glycoforms 384:409 undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) 384:466 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 0 85 theme serum 22:26 arg1 O-glycosylation 28:42 serum O-glycosylation 22:42 serum O-glycosylation 22:42 MALDI-MS profiling of serum O-glycosylation and N-glycosylation in COG5-CDG. 28444691 4 86 theme multisystem 584:594 arg1 disease 596:602 a multisystem disease 582:602 a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior 582:743 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 86 theme multisystem 584:594 arg1 COG5-CDG 544:551 COG5-CDG 544:551 COG5-CDG (COG5 subunit deficiency) 544:577 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 2 87 theme Golgi 197:201 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 87 theme Golgi 197:201 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 5 88 theme differential 839:850 arg1 O-glycoform 858:868 differential serum O-glycoform 839:868 differential serum O-glycoform 839:868 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 6 89 theme age-matched 937:947 arg1 controls 949:956 age-matched controls 937:956 age-matched controls 937:956 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 4 90 theme COG5 554:557 arg1 deficiency 567:576 COG5 subunit deficiency 554:576 COG5 subunit deficiency 554:576 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 4 90 theme COG5 554:557 arg1 COG5-CDG 544:551 COG5-CDG 544:551 COG5-CDG (COG5 subunit deficiency) 544:577 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 2 91 theme Conserved 176:184 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 91 theme Conserved 176:184 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 5 92 gly N-glycoform 874:884 arg2 patients 894:901 five patients 889:901 five patients with COG5-CDG 889:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 7 93 theme glycans 1263:1269 arg1 increase 1289:1296 slight increase 1282:1296 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 7 93 theme glycans 1263:1269 arg1 abundance 1181:1189 the relative abundance 1168:1189 the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans 1168:1342 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 28444691 3 94 theme serum 414:418 arg1 transferrin 420:430 serum transferrin 414:430 serum transferrin 414:430 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 6 95 gly mono-sialylated 1058:1072 arg1 apoC-III1 1047:1055 apoC-III1 1047:1055 apoC-III1 (mono-sialylated glycoform) 1047:1083 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 6 95 gly mono-sialylated 1058:1072 arg1 glycoform 1074:1082 mono-sialylated glycoform 1058:1082 mono-sialylated glycoform 1058:1082 When compared with age-matched controls, COG5-CDG showed a significant increase of apoC-III0a (aglycosylated glycoform), whereas apoC-III1 (mono-sialylated glycoform) decreased significantly. 28444691 4 96 theme intellectual 630:641 arg1 disability 643:652 intellectual disability 630:652 intellectual disability of variable degree 630:671 COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior. 28444691 3 97 dep focusing 361:368 arg1 isoelectric 349:359 isoelectric 349:359 isoelectric 349:359 In COG-CDG, isoelectric focusing separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N-glycosylation and O-glycosylation defects, respectively. 28444691 2 98 theme COG 204:206 arg1 diseases 225:232 genetic diseases 217:232 genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities 217:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 98 theme COG 204:206 arg1 -CDG 208:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG 176:211 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 2 99 theme processing 311:320 arg1 abnormalities 322:334 O-glycan processing abnormalities 302:334 O-glycan processing abnormalities 302:334 Conserved oligomeric Golgi (COG)-CDG are genetic diseases due to defects of the COG complex subunits 1-8 causing N-glycan and O-glycan processing abnormalities. 28444691 5 100 gly O-glycoform 858:868 arg2 patients 894:901 five patients 889:901 five patients with COG5-CDG 889:915 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 5 100 gly O-glycoform 858:868 arg1 serum 852:856 differential serum O-glycoform 839:868 differential serum O-glycoform 839:868 We applied matrix-assisted laser desorption/ionization-MS for a high-throughput screening of differential serum O-glycoform and N-glycoform in five patients with COG5-CDG. 28444691 7 101 theme high-mannose 1301:1312 arg1 structures 1314:1323 high-mannose structures 1301:1323 high-mannose structures 1301:1323 Serum N-glycome of COG5-CDG patients was characterized by the relative abundance of undersialylated and undergalactosylated biantennary and triantennary glycans as well as slight increase of high-mannose structures and hybrid glycans. 27529638 5 0 theme eukaryotic 828:837 arg1 N-glycans 839:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 1 1 theme peptide/protein 175:189 arg1 glycosylation 158:170 site-directed glycosylation 144:170 site-directed glycosylation of peptide/protein 144:189 Here we report a facile and efficient method for site-directed glycosylation of peptide/protein. 27529638 3 2 theme pharmaceutical 349:362 arg1 peptide 364:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 2 theme pharmaceutical 349:362 arg1 peptide-1 387:395 glucagon-like peptide-1 373:395 glucagon-like peptide-1 (GLP-1) 373:403 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 5 3 gly glycopeptide/glycoprotein 794:818 arg1 glycopeptide/glycoprotein 794:818 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 2 4 theme N-glycan 316:323 arg1 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 4 theme N-glycan 316:323 arg1 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 4 theme N-glycan 316:323 arg1 generation 234:243 generation 234:243 generation of a GlcNAc-O-peptide/protein 234:273 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 3 5 theme glucagon-like 373:385 arg1 peptide 364:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 5 theme glucagon-like 373:385 arg1 peptide-1 387:395 glucagon-like peptide-1 373:395 glucagon-like peptide-1 (GLP-1) 373:403 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 5 theme glucagon-like 373:385 arg1 GLP-1 398:402 GLP-1 398:402 GLP-1 398:402 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 4 6 with GLP-1 525:529 arg1 N-glycan 545:552 O-linked N-glycan 536:552 O-linked N-glycan 536:552 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 2 7 theme eukaryotic 305:314 arg1 N-glycan 316:323 a eukaryotic N-glycan 303:323 a eukaryotic N-glycan to the GlcNAc moiety 303:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 5 8 theme homogeneous 782:792 arg1 N-glycans 839:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 4 9 link O-linked 536:543 arg1 N-glycan 545:552 O-linked N-glycan 536:552 O-linked N-glycan 536:552 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 4 10 theme drug 693:696 arg1 production 698:707 peptide/protein drug production 677:707 peptide/protein drug production 677:707 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 2 11 contain contains 203:210 arg2 generation 234:243 generation 234:243 generation of a GlcNAc-O-peptide/protein 234:273 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 11 contain contains 203:210 arg2 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 11 contain contains 203:210 arg2 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 11 contain contains 203:210 arg1 method 196:201 The method 192:201 The method 192:201 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 0 12 theme Site-Directed 0:12 arg1 Glycosylation 14:26 Site-Directed Glycosylation 0:26 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 0:93 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 4 13 theme unchanged 568:576 arg1 structure 588:596 an unchanged secondary structure 565:596 an unchanged secondary structure 565:596 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 0 14 theme Peptide/Protein 31:45 arg1 Glycosylation 14:26 Site-Directed Glycosylation 0:26 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 0:93 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 2 15 theme GlcNAc 332:337 arg1 moiety 339:344 the GlcNAc moiety 328:344 the GlcNAc moiety 328:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 4 16 theme approach 664:671 arg1 application 644:654 the potential application 630:654 the potential application of this approach for peptide/protein drug production 630:707 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 0 17 theme Homogeneous 52:62 arg1 N-Glycans 84:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 4 18 theme secondary 578:586 arg1 structure 588:596 an unchanged secondary structure 565:596 an unchanged secondary structure 565:596 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 4 19 theme peptide/protein 677:691 arg1 production 698:707 peptide/protein drug production 677:707 peptide/protein drug production 677:707 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 3 20 theme bovine 427:432 arg1 protein 418:424 a model protein 410:424 a model protein 410:424 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 20 theme bovine 427:432 arg1 α-Crystallin 434:445 bovine α-Crystallin 427:445 bovine α-Crystallin 427:445 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 4 21 theme O-linked 536:543 arg1 N-glycan 545:552 O-linked N-glycan 536:552 O-linked N-glycan 536:552 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 27529638 5 22 theme general 754:760 arg1 strategy 762:769 a general strategy 752:769 a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 752:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 0 23 gly Glycosylation 14:26 arg1 Peptide/Protein 31:45 Peptide/Protein 31:45 Peptide/Protein 31:45 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 0 24 theme Eukaryotic 73:82 arg1 N-Glycans 84:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 5 25 theme bearing 820:826 arg1 N-glycans 839:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 1 26 theme efficient 123:131 arg1 method 133:138 a facile and efficient method 110:138 a facile and efficient method for site-directed glycosylation of peptide/protein 110:189 Here we report a facile and efficient method for site-directed glycosylation of peptide/protein. 27529638 0 27 theme O-Linked 64:71 arg1 N-Glycans 84:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 1 28 gly glycosylation 158:170 arg1 peptide/protein 175:189 peptide/protein 175:189 peptide/protein 175:189 Here we report a facile and efficient method for site-directed glycosylation of peptide/protein. 27529638 3 29 gly glycosylated 466:477 arg1 peptide 364:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide 347:370 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 29 gly glycosylated 466:477 arg1 protein 418:424 a model protein 410:424 a model protein 410:424 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 29 gly glycosylated 466:477 arg1 α-Crystallin 434:445 bovine α-Crystallin 427:445 bovine α-Crystallin 427:445 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 29 gly glycosylated 466:477 arg1 peptide-1 387:395 glucagon-like peptide-1 373:395 glucagon-like peptide-1 (GLP-1) 373:403 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 0 30 with Glycosylation 14:26 arg1 N-Glycans 84:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Homogeneous O-Linked Eukaryotic N-Glycans 52:92 Site-Directed Glycosylation of Peptide/Protein with Homogeneous O-Linked Eukaryotic N-Glycans. 27529638 2 31 theme GlcNAc-O-peptide/protein 250:273 arg1 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 31 theme GlcNAc-O-peptide/protein 250:273 arg1 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 31 theme GlcNAc-O-peptide/protein 250:273 arg1 generation 234:243 generation 234:243 generation of a GlcNAc-O-peptide/protein 234:273 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 1 32 theme facile 112:117 arg1 method 133:138 a facile and efficient method 110:138 a facile and efficient method for site-directed glycosylation of peptide/protein 110:189 Here we report a facile and efficient method for site-directed glycosylation of peptide/protein. 27529638 2 33 dep steps 227:231 arg1 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 33 dep steps 227:231 arg1 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 33 dep steps 227:231 arg1 generation 234:243 generation 234:243 generation of a GlcNAc-O-peptide/protein 234:273 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 5 34 theme coupled 726:732 arg1 approach 734:741 the coupled approach 722:741 the coupled approach 722:741 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 1 35 theme site-directed 144:156 arg1 glycosylation 158:170 site-directed glycosylation 144:170 site-directed glycosylation of peptide/protein 144:189 Here we report a facile and efficient method for site-directed glycosylation of peptide/protein. 27529638 5 36 theme glycopeptide/glycoprotein 794:818 arg1 N-glycans 839:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans 782:847 In summary, the coupled approach provides a general strategy to produce homogeneous glycopeptide/glycoprotein bearing eukaryotic N-glycans. 27529638 2 37 theme sequential 216:225 arg1 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 37 theme sequential 216:225 arg1 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 37 theme sequential 216:225 arg1 generation 234:243 generation 234:243 generation of a GlcNAc-O-peptide/protein 234:273 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 38 theme subsequent 280:289 arg1 steps 227:231 two sequential steps 212:231 two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 212:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 2 38 theme subsequent 280:289 arg1 ligation 291:298 subsequent ligation 280:298 subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety 280:344 The method contains two sequential steps: generation of a GlcNAc-O-peptide/protein, and subsequent ligation of a eukaryotic N-glycan to the GlcNAc moiety. 27529638 3 39 theme model 412:416 arg1 protein 418:424 a model protein 410:424 a model protein 410:424 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 3 39 theme model 412:416 arg1 α-Crystallin 434:445 bovine α-Crystallin 427:445 bovine α-Crystallin 427:445 A pharmaceutical peptide, glucagon-like peptide-1 (GLP-1), and a model protein, bovine α-Crystallin, were successfully glycosylated using such an approach. 27529638 4 40 theme potential 634:642 arg1 application 644:654 the potential application 630:654 the potential application of this approach for peptide/protein drug production 630:707 It was shown that the GLP-1 with O-linked N-glycan maintained an unchanged secondary structure after glycosylation, suggesting the potential application of this approach for peptide/protein drug production. 28409838 4 0 theme constant 853:860 arg1 profile 876:882 a constant glycosylation profile 851:882 a constant glycosylation profile 851:882 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 10 1 theme model 2058:2062 arg1 estimation 2074:2083 the model parameter estimation 2054:2083 the model parameter estimation 2054:2083 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 3 2 contain has 603:605 arg2 potential 611:619 the potential to increase therapeutic efficacy and bioactivity 607:668 the potential to increase therapeutic efficacy and bioactivity 607:668 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 3 2 contain has 603:605 arg1 production 543:552 the production 539:552 the production of a confined mAb N-linked glycosylation pattern 539:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 4 3 theme profile 876:882 arg1 production 837:846 the production 833:846 the production of a constant glycosylation profile 833:882 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 6 4 theme different 1156:1164 arg1 concentrations 1207:1220 different viable cell density and media supplement concentrations 1156:1220 different viable cell density and media supplement concentrations 1156:1220 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 11 5 theme rational 2254:2261 arg1 strategy 2263:2270 an efficient and rational strategy 2237:2270 an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns 2237:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 11 6 theme glycosylation 2136:2148 arg1 modulation 2122:2131 the modulation 2118:2131 the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model 2118:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 9 7 theme obtained 1657:1664 arg1 data 1679:1682 The obtained experimental data 1653:1682 The obtained experimental data set 1653:1686 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 0 8 link N-linked 47:54 arg1 glycosylation 56:68 monoclonal antibody N-linked glycosylation 27:68 monoclonal antibody N-linked glycosylation 27:68 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 11 9 theme efficient 2240:2248 arg1 strategy 2263:2270 an efficient and rational strategy 2237:2270 an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns 2237:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 2 10 theme state 316:320 arg1 operation 322:330 The steady state operation 305:330 The steady state operation at high viable cell density 305:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 1 11 theme fed-batch 204:212 arg1 processes 214:222 traditional fed-batch processes 192:222 traditional fed-batch processes 192:222 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 1 11 theme fed-batch 204:212 arg1 antibodies 287:296 monoclonal antibodies 276:296 monoclonal antibodies (mAb) 276:302 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 6 12 theme Experiment 1135:1144 arg1 Design 1125:1130 a Design 1123:1130 a Design of Experiment (DoE) 1123:1150 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 11 13 theme continuous 2283:2292 arg1 processes 2294:2302 continuous processes 2283:2302 continuous processes with desired N-linked glycosylation patterns 2283:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 3 14 theme mAb 568:570 arg1 pattern 595:601 a confined mAb N-linked glycosylation pattern 557:601 a confined mAb N-linked glycosylation pattern 557:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 8 15 theme high 1422:1425 arg1 VCD 1450:1452 VCD 1450:1452 VCD 1450:1452 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 15 theme high 1422:1425 arg1 densities 1439:1447 high viable cell densities 1422:1447 high viable cell densities (VCD) values 1422:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 9 16 link N-linked 1796:1803 arg1 glycosylation 1805:1817 N-linked glycosylation 1796:1817 N-linked glycosylation 1796:1817 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 10 17 theme glycosylation 1965:1977 arg1 pattern 1979:1985 glycosylation pattern 1965:1985 glycosylation pattern 1965:1985 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 15 18 dep © 2391:2391 arg1 Inc. 2417:2420 Inc. 2417:2420 Inc. 2417:2420 © 2017 Wiley Periodicals, Inc. 28409838 8 19 theme cell 1434:1437 arg1 VCD 1450:1452 VCD 1450:1452 VCD 1450:1452 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 19 theme cell 1434:1437 arg1 densities 1439:1447 high viable cell densities 1422:1447 high viable cell densities (VCD) values 1422:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 6 20 theme supplement 1196:1205 arg1 concentrations 1207:1220 different viable cell density and media supplement concentrations 1156:1220 different viable cell density and media supplement concentrations 1156:1220 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 4 21 theme flow 719:722 arg1 rates 724:728 flow rates 719:728 flow rates 719:728 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 5 22 theme intracellular 1016:1028 arg1 processes 1030:1038 extra- and intracellular processes 1005:1038 extra- and intracellular processes 1005:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 5 23 theme Steady 902:907 arg1 state 909:913 Steady state 902:913 Steady state 902:913 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 11 24 from approach 2179:2186 arg1 combination 2191:2201 combination 2191:2201 combination with mechanistic model 2191:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 10 25 dep pattern 1979:1985 arg1 i.e. 1959:1962 i.e. 1959:1962 i.e. 1959:1962 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 6 26 theme cell 1173:1176 arg1 density 1178:1184 viable cell density 1166:1184 viable cell density 1166:1184 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 3 27 link N-linked 572:579 arg1 pattern 595:601 a confined mAb N-linked glycosylation pattern 557:601 a confined mAb N-linked glycosylation pattern 557:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 0 28 from modeling 15:22 arg1 reactors 98:105 mammalian cell perfusion reactors 73:105 mammalian cell perfusion reactors 73:105 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 0 29 theme cell 83:86 arg1 reactors 98:105 mammalian cell perfusion reactors 73:105 mammalian cell perfusion reactors 73:105 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 9 30 theme surface 1750:1756 arg1 model 1758:1762 a statistical response surface model 1727:1762 a statistical response surface model (RSM) 1727:1768 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 9 30 theme surface 1750:1756 arg1 RSM 1765:1767 RSM 1765:1767 RSM 1765:1767 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 0 31 from Modulation 0:9 arg1 reactors 98:105 mammalian cell perfusion reactors 73:105 mammalian cell perfusion reactors 73:105 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 1 32 theme cell 118:121 arg1 cultures 133:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 8 33 theme complex 1490:1496 arg1 structures 1505:1514 complex glycan structures 1490:1514 complex glycan structures 1490:1514 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 4 34 theme Chinese 771:777 arg1 CHO 794:796 CHO 794:796 CHO 794:796 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 4 34 theme Chinese 771:777 arg1 ovary 787:791 Chinese hamster ovary 771:791 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 35 theme product 403:409 arg1 delivery 382:389 the continuous delivery 367:389 the continuous delivery of antibody product 367:409 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 11 36 theme sequential 2155:2164 arg1 approach 2179:2186 a sequential steady state approach 2153:2186 a sequential steady state approach in combination with mechanistic model 2153:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 4 37 theme composition 737:747 arg1 control 708:714 accurate control 699:714 accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 699:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 38 theme reduced 447:453 arg1 variability 466:476 reduced in-process variability 447:476 reduced in-process variability 447:476 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 4 39 theme cell 753:756 arg1 density 758:764 cell density 753:764 cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 753:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 5 40 theme initial 936:942 arg1 phase 955:959 an initial transition phase 933:959 an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes 933:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 7 41 theme various 1295:1301 arg1 states 1310:1315 various steady states 1295:1315 various steady states 1295:1315 This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. 28409838 2 42 theme quality 498:504 arg1 CQA 518:520 CQA 518:520 CQA 518:520 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 2 42 theme quality 498:504 arg1 attributes 506:515 critical product quality attributes 481:515 critical product quality attributes (CQA) 481:521 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 4 43 theme cell 799:802 arg1 bioreactor 814:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 44 theme increased 416:424 arg1 yield 437:441 increased space-time yield 416:441 increased space-time yield 416:441 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 0 45 theme monoclonal 27:36 arg1 glycosylation 56:68 monoclonal antibody N-linked glycosylation 27:68 monoclonal antibody N-linked glycosylation 27:68 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 11 46 theme steady 2166:2171 arg1 approach 2179:2186 a sequential steady state approach 2153:2186 a sequential steady state approach in combination with mechanistic model 2153:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 2 47 theme critical 481:488 arg1 CQA 518:520 CQA 518:520 CQA 518:520 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 2 47 theme critical 481:488 arg1 attributes 506:515 critical product quality attributes 481:515 critical product quality attributes (CQA) 481:521 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 8 48 theme galactose 1547:1555 arg1 supplementation 1521:1535 the supplementation 1517:1535 the supplementation of either galactose, or manganese as well as their synergy 1517:1594 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 48 theme galactose 1547:1555 arg1 synergy 1588:1594 their synergy 1582:1594 the supplementation of either galactose, or manganese as well as their synergy 1517:1594 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 4 49 theme bioreactor 814:823 arg1 rates 724:728 flow rates 719:728 flow rates 719:728 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 4 49 theme bioreactor 814:823 arg1 composition 737:747 media composition 731:747 media composition 731:747 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 4 49 theme bioreactor 814:823 arg1 density 758:764 cell density 753:764 cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 753:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 50 theme cell 347:350 arg1 density 352:358 high viable cell density 335:358 high viable cell density 335:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 0 51 theme N-linked 47:54 arg1 glycosylation 56:68 monoclonal antibody N-linked glycosylation 27:68 monoclonal antibody N-linked glycosylation 27:68 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 9 52 used used 1692:1695 arg2 data 1679:1682 The obtained experimental data 1653:1682 The obtained experimental data set 1653:1686 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 7 53 theme screening 1269:1277 arg1 approach 1279:1286 a sequential screening approach 1256:1286 a sequential screening approach 1256:1286 This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. 28409838 11 54 theme desired 2309:2315 arg1 patterns 2340:2347 desired N-linked glycosylation patterns 2309:2347 desired N-linked glycosylation patterns 2309:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 11 55 with combination 2191:2201 arg1 model 2220:2224 mechanistic model 2208:2224 mechanistic model 2208:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 10 56 theme surface 1857:1863 arg1 predictions 1865:1875 the response surface predictions 1844:1875 the response surface predictions with respect to its capability and reliability 1844:1922 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 4 57 theme ovary 787:791 arg1 bioreactor 814:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 58 theme high 335:338 arg1 density 352:358 high viable cell density 335:358 high viable cell density 335:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 10 59 theme DoE 2034:2036 arg1 design 2038:2043 the DoE design 2030:2043 the DoE design used for the model parameter estimation 2030:2083 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 11 60 link N-linked 2317:2324 arg1 patterns 2340:2347 desired N-linked glycosylation patterns 2309:2347 desired N-linked glycosylation patterns 2309:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 2 61 theme continuous 371:380 arg1 delivery 382:389 the continuous delivery 367:389 the continuous delivery of antibody product 367:409 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 8 62 theme complex 1638:1644 arg1 forms 1646:1650 complex forms 1638:1650 complex forms 1638:1650 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 1 63 theme proteins 258:265 arg1 production 232:241 the production 228:241 the production of therapeutic proteins 228:265 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 9 64 theme N-linked 1796:1803 arg1 glycosylation 1805:1817 N-linked glycosylation 1796:1817 N-linked glycosylation 1796:1817 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 10 65 theme parameter 2064:2072 arg1 estimation 2074:2083 the model parameter estimation 2054:2083 the model parameter estimation 2054:2083 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 9 66 theme statistical 1729:1739 arg1 model 1758:1762 a statistical response surface model 1727:1762 a statistical response surface model (RSM) 1727:1768 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 9 66 theme statistical 1729:1739 arg1 RSM 1765:1767 RSM 1765:1767 RSM 1765:1767 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 6 67 theme glycosylation 1073:1085 arg1 profile 1087:1093 the glycosylation profile 1069:1093 the glycosylation profile 1069:1093 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 11 68 theme glycosylation 2326:2338 arg1 patterns 2340:2347 desired N-linked glycosylation patterns 2309:2347 desired N-linked glycosylation patterns 2309:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 2 69 from density 352:358 arg1 operation 322:330 The steady state operation 305:330 The steady state operation at high viable cell density 305:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 8 70 theme high 1391:1394 arg1 levels 1404:1409 high ammonia levels 1391:1409 high ammonia levels reached at high viable cell densities (VCD) values 1391:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 1 71 theme monoclonal 276:285 arg1 mAb 299:301 mAb 299:301 mAb 299:301 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 1 71 theme monoclonal 276:285 arg1 antibodies 287:296 monoclonal antibodies 276:296 monoclonal antibodies (mAb) 276:302 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 11 72 with processes 2294:2302 arg1 patterns 2340:2347 desired N-linked glycosylation patterns 2309:2347 desired N-linked glycosylation patterns 2309:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 4 73 theme glycosylation 862:874 arg1 profile 876:882 a constant glycosylation profile 851:882 a constant glycosylation profile 851:882 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 11 74 from modulation 2122:2131 arg1 approach 2179:2186 a sequential steady state approach 2153:2186 a sequential steady state approach in combination with mechanistic model 2153:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 9 75 theme response 1741:1748 arg1 model 1758:1762 a statistical response surface model 1727:1762 a statistical response surface model (RSM) 1727:1768 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 9 75 theme response 1741:1748 arg1 RSM 1765:1767 RSM 1765:1767 RSM 1765:1767 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 3 76 theme N-linked 572:579 arg1 pattern 595:601 a confined mAb N-linked glycosylation pattern 557:601 a confined mAb N-linked glycosylation pattern 557:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 1 77 theme traditional 192:202 arg1 processes 214:222 traditional fed-batch processes 192:222 traditional fed-batch processes 192:222 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 1 77 theme traditional 192:202 arg1 antibodies 287:296 monoclonal antibodies 276:296 monoclonal antibodies (mAb) 276:302 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 9 78 theme experimental 1666:1677 arg1 data 1679:1682 The obtained experimental data 1653:1682 The obtained experimental data set 1653:1686 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 10 79 theme system 1942:1947 arg1 behavior 1949:1956 the system behavior 1938:1956 the system behavior (i.e., glycosylation pattern) 1938:1986 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 3 80 theme confined 559:566 arg1 pattern 595:601 a confined mAb N-linked glycosylation pattern 557:601 a confined mAb N-linked glycosylation pattern 557:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 3 81 theme glycosylation 581:593 arg1 pattern 595:601 a confined mAb N-linked glycosylation pattern 557:601 a confined mAb N-linked glycosylation pattern 557:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 8 82 theme viable 1427:1432 arg1 VCD 1450:1452 VCD 1450:1452 VCD 1450:1452 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 82 theme viable 1427:1432 arg1 densities 1439:1447 high viable cell densities 1422:1447 high viable cell densities (VCD) values 1422:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 83 theme densities 1439:1447 arg1 values 1455:1460 high viable cell densities (VCD) values 1422:1460 high viable cell densities (VCD) values 1422:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 3 84 theme pattern 595:601 arg1 production 543:552 the production 539:552 the production of a confined mAb N-linked glycosylation pattern 539:601 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 1 85 theme perfusion 123:131 arg1 cultures 133:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 6 86 theme media 1190:1194 arg1 concentrations 1207:1220 different viable cell density and media supplement concentrations 1156:1220 different viable cell density and media supplement concentrations 1156:1220 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 5 87 theme processes 1030:1038 arg1 stabilization 988:1000 the stabilization 984:1000 the stabilization of extra- and intracellular processes 984:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 3 88 theme therapeutic 633:643 arg1 efficacy 645:652 therapeutic efficacy 633:652 therapeutic efficacy 633:652 In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. 28409838 2 89 theme steady 309:314 arg1 operation 322:330 The steady state operation 305:330 The steady state operation at high viable cell density 305:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 6 90 theme density 1178:1184 arg1 concentrations 1207:1220 different viable cell density and media supplement concentrations 1156:1220 different viable cell density and media supplement concentrations 1156:1220 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 4 91 theme rates 724:728 arg1 control 708:714 accurate control 699:714 accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 699:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 0 92 theme mammalian 73:81 arg1 reactors 98:105 mammalian cell perfusion reactors 73:105 mammalian cell perfusion reactors 73:105 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 1 93 theme renewed 154:160 arg1 interest 162:169 renewed interest 154:169 renewed interest 154:169 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 1 93 theme renewed 154:160 arg1 alternative 177:187 an alternative 174:187 an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb) 174:302 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 11 94 theme state 2173:2177 arg1 approach 2179:2186 a sequential steady state approach 2153:2186 a sequential steady state approach in combination with mechanistic model 2153:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 6 95 theme viable 1166:1171 arg1 density 1178:1184 viable cell density 1166:1184 viable cell density 1166:1184 The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. 28409838 0 96 theme perfusion 88:96 arg1 reactors 98:105 mammalian cell perfusion reactors 73:105 mammalian cell perfusion reactors 73:105 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 2 97 theme attributes 506:515 arg1 variability 466:476 reduced in-process variability 447:476 reduced in-process variability 447:476 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 2 97 theme attributes 506:515 arg1 yield 437:441 increased space-time yield 416:441 increased space-time yield 416:441 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 4 98 theme accurate 699:706 arg1 control 708:714 accurate control 699:714 accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 699:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 4 99 theme hamster 779:785 arg1 CHO 794:796 CHO 794:796 CHO 794:796 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 4 99 theme hamster 779:785 arg1 ovary 787:791 Chinese hamster ovary 771:791 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 5 100 theme transition 944:953 arg1 phase 955:959 an initial transition phase 933:959 an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes 933:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 9 101 theme model 1758:1762 arg1 reliability 1712:1722 the reliability 1708:1722 the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation 1708:1817 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 8 102 theme glycan 1498:1503 arg1 structures 1505:1514 complex glycan structures 1490:1514 complex glycan structures 1490:1514 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 4 103 theme density 758:764 arg1 control 708:714 accurate control 699:714 accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor 699:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 10 104 theme experimental 2000:2011 arg1 space 2013:2017 the experimental space 1996:2017 the experimental space covered by the DoE design used for the model parameter estimation 1996:2083 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 11 105 theme mechanistic 2208:2218 arg1 model 2220:2224 mechanistic model 2208:2224 mechanistic model 2208:2224 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 2 106 theme antibody 394:401 arg1 product 403:409 antibody product 394:409 antibody product 394:409 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 7 107 theme steady 1303:1308 arg1 states 1310:1315 various steady states 1295:1315 various steady states 1295:1315 This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. 28409838 2 108 theme in-process 455:464 arg1 variability 466:476 reduced in-process variability 447:476 reduced in-process variability 447:476 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 14 109 dep 1978-1990 2380:2388 arg1 2017 2370:2373 2017 2370:2373 2017 2370:2373 2017;114: 1978-1990. 28409838 4 110 theme media 731:735 arg1 composition 737:747 media composition 731:747 media composition 731:747 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 1 111 theme Mammalian 108:116 arg1 cultures 133:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures 108:140 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 28409838 9 112 theme mechanistic 1775:1785 arg1 model 1787:1791 a mechanistic model 1773:1791 a mechanistic model of N-linked glycosylation 1773:1817 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 2 113 theme space-time 426:435 arg1 yield 437:441 increased space-time yield 416:441 increased space-time yield 416:441 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 0 114 theme antibody 38:45 arg1 glycosylation 56:68 monoclonal antibody N-linked glycosylation 27:68 monoclonal antibody N-linked glycosylation 27:68 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 2 115 theme product 490:496 arg1 CQA 518:520 CQA 518:520 CQA 518:520 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 2 115 theme product 490:496 arg1 attributes 506:515 critical product quality attributes 481:515 critical product quality attributes (CQA) 481:521 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 0 116 theme glycosylation 56:68 arg1 modeling 15:22 modeling 15:22 modeling 15:22 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 0 116 theme glycosylation 56:68 arg1 Modulation 0:9 Modulation 0:9 Modulation 0:9 Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors. 28409838 5 117 theme extra- 1005:1010 arg1 processes 1030:1038 extra- and intracellular processes 1005:1038 extra- and intracellular processes 1005:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 7 118 theme sequential 1258:1267 arg1 approach 1279:1286 a sequential screening approach 1256:1286 a sequential screening approach 1256:1286 This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. 28409838 9 119 theme glycosylation 1805:1817 arg1 model 1787:1791 a mechanistic model 1773:1791 a mechanistic model of N-linked glycosylation 1773:1817 The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. 28409838 8 120 theme manganese 1561:1569 arg1 supplementation 1521:1535 the supplementation 1517:1535 the supplementation of either galactose, or manganese as well as their synergy 1517:1594 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 120 theme manganese 1561:1569 arg1 synergy 1588:1594 their synergy 1582:1594 the supplementation of either galactose, or manganese as well as their synergy 1517:1594 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 4 121 theme perfusion 804:812 arg1 bioreactor 814:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 a Chinese hamster ovary (CHO) cell perfusion bioreactor 769:823 In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. 28409838 2 122 theme viable 340:345 arg1 density 352:358 high viable cell density 335:358 high viable cell density 335:358 The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). 28409838 5 123 theme days 966:969 arg1 phase 955:959 an initial transition phase 933:959 an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes 933:1038 Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. 28409838 11 124 theme N-linked 2317:2324 arg1 patterns 2340:2347 desired N-linked glycosylation patterns 2309:2347 desired N-linked glycosylation patterns 2309:2347 Therefore, we can conclude that the modulation of glycosylation in a sequential steady state approach in combination with mechanistic model represents an efficient and rational strategy to develop continuous processes with desired N-linked glycosylation patterns. 28409838 10 125 theme response 1848:1855 arg1 predictions 1865:1875 the response surface predictions 1844:1875 the response surface predictions with respect to its capability and reliability 1844:1922 The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. 28409838 8 126 theme ammonia 1396:1402 arg1 levels 1404:1409 high ammonia levels 1391:1409 high ammonia levels reached at high viable cell densities (VCD) values 1391:1460 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 8 127 theme forms 1646:1650 arg1 proportion 1624:1633 the proportion 1620:1633 the proportion of complex forms 1620:1650 It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. 28409838 1 128 theme therapeutic 246:256 arg1 proteins 258:265 therapeutic proteins 246:265 therapeutic proteins 246:265 Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). 26267136 6 0 theme dimer-bridged 868:880 arg1 conformations 849:861 these conformations 843:861 these conformations 843:861 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 6 0 theme dimer-bridged 868:880 arg1 one 915:917 one 915:917 one 915:917 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 6 0 theme dimer-bridged 868:880 arg1 146-N-glycan 882:893 the dimer-bridged 146-N-glycan 864:893 the dimer-bridged 146-N-glycan 864:893 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 4 1 theme crystal 622:628 arg1 structures 630:639 crystal structures 622:639 crystal structures of NAs 622:646 Moreover, with 146-N-glycan, our simulation result was more consistent with crystal structures of NAs than simulations conducted without glycans. 26267136 6 2 theme 150-loop 959:966 arg1 conformation 968:979 the 150-loop conformation 955:979 the 150-loop conformation 955:979 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 0 3 theme Neuraminidase 63:75 arg1 Neuraminidase 63:75 Neuraminidase 63:75 Neuraminidase 63:75 The Mechanism by which 146-N-Glycan Affects the Active Site of Neuraminidase. 26267136 0 3 theme Neuraminidase 63:75 arg1 Site 55:58 the Active Site 44:58 the Active Site of Neuraminidase 44:75 The Mechanism by which 146-N-Glycan Affects the Active Site of Neuraminidase. 26267136 7 4 theme binding 1103:1109 arg1 pocket 1111:1116 the binding pocket 1099:1116 the binding pocket 1099:1116 Furthermore, our simulation revealed that various standing conformations of 146-N-glycan could block the entrance of the binding pocket. 26267136 2 5 theme drug 288:291 arg1 development 293:303 rational drug development 279:303 rational drug development against the antiviral resistance of influenza 279:349 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 8 6 with inhibitors 1214:1223 arg1 substituents 1239:1250 flexible substituents 1230:1250 flexible substituents 1230:1250 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 6 7 theme conformations 849:861 arg1 conformations 849:861 these conformations 843:861 these conformations 843:861 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 6 7 theme conformations 849:861 arg1 one 915:917 one 915:917 one 915:917 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 6 7 theme conformations 849:861 arg1 146-N-glycan 882:893 the dimer-bridged 146-N-glycan 864:893 the dimer-bridged 146-N-glycan 864:893 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 26267136 2 8 theme active 236:241 arg1 site 243:246 the active site 232:246 the active site 232:246 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 8 9 theme experimental 1151:1162 arg1 data 1164:1167 experimental data 1151:1167 experimental data 1151:1167 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 2 10 theme rational 279:286 arg1 development 293:303 rational drug development 279:303 rational drug development against the antiviral resistance of influenza 279:349 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 2 11 theme influenza 341:349 arg1 resistance 327:336 the antiviral resistance 313:336 the antiviral resistance of influenza 313:349 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 7 12 theme pocket 1111:1116 arg1 entrance 1087:1094 the entrance 1083:1094 the entrance of the binding pocket 1083:1116 Furthermore, our simulation revealed that various standing conformations of 146-N-glycan could block the entrance of the binding pocket. 26267136 4 13 with consistent 606:615 arg1 146-N-glycan 561:572 146-N-glycan 561:572 146-N-glycan 561:572 Moreover, with 146-N-glycan, our simulation result was more consistent with crystal structures of NAs than simulations conducted without glycans. 26267136 4 13 with consistent 606:615 arg1 structures 630:639 crystal structures 622:639 crystal structures of NAs 622:646 Moreover, with 146-N-glycan, our simulation result was more consistent with crystal structures of NAs than simulations conducted without glycans. 26267136 5 14 theme dimer-bridged 812:824 arg1 conformations 780:792 three distinct conformations 765:792 three distinct conformations: monomer-bridged, dimer-bridged and standing 765:837 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 7 15 theme 146-N-glycan 1058:1069 arg1 conformations 1041:1053 various standing conformations 1024:1053 various standing conformations of 146-N-glycan 1024:1069 Furthermore, our simulation revealed that various standing conformations of 146-N-glycan could block the entrance of the binding pocket. 26267136 3 16 theme 150-loop 493:500 arg1 conformation 473:484 the conformation 469:484 the conformation of the 150-loop that controls the volume of the 150-cavity 469:543 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 8 17 with consistent 1135:1144 arg1 data 1164:1167 experimental data 1151:1167 experimental data 1151:1167 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 2 18 theme antiviral 317:325 arg1 resistance 327:336 the antiviral resistance 313:336 the antiviral resistance of influenza 313:349 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 9 19 theme rigid 1325:1329 arg1 inhibitors 1382:1391 better inhibitors 1375:1391 better inhibitors targeting the 150-cavity 1375:1416 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 9 19 theme rigid 1325:1329 arg1 substituents 1347:1358 rigid and hydrophobic substituents 1325:1358 rigid and hydrophobic substituents 1325:1358 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 1 20 gly glycosylation 104:116 arg1 NA 142:143 NA 142:143 NA 142:143 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 20 gly glycosylation 104:116 arg2 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 20 gly glycosylation 104:116 arg1 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 20 gly glycosylation 104:116 arg2 sites 118:122 the most conserved glycosylation sites 85:122 the most conserved glycosylation sites of neuraminidase (NA) 85:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 5 21 theme trajectories 719:730 arg1 analysis 700:707 Cluster analysis 692:707 Cluster analysis of the MD trajectories 692:730 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 3 22 theme dynamics 394:401 arg1 simulations 408:418 2.4 μs molecular dynamics (MD) simulations 377:418 2.4 μs molecular dynamics (MD) simulations 377:418 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 8 23 theme low 1198:1200 arg1 activity 1202:1209 the relatively low activity 1183:1209 the relatively low activity of inhibitors with flexible substituents toward the 150-cavity 1183:1272 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 9 24 theme hydrophobic 1335:1345 arg1 inhibitors 1382:1391 better inhibitors 1375:1391 better inhibitors targeting the 150-cavity 1375:1416 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 9 24 theme hydrophobic 1335:1345 arg1 substituents 1347:1358 rigid and hydrophobic substituents 1325:1358 rigid and hydrophobic substituents 1325:1358 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 7 25 theme various 1024:1030 arg1 conformations 1041:1053 various standing conformations 1024:1053 various standing conformations of 146-N-glycan 1024:1069 Furthermore, our simulation revealed that various standing conformations of 146-N-glycan could block the entrance of the binding pocket. 26267136 2 26 theme NA 206:207 arg1 150-cavity 192:201 the 150-cavity 188:201 the 150-cavity of NA, which is found within the active site 188:246 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 5 27 theme standing 830:837 arg1 conformations 780:792 three distinct conformations 765:792 three distinct conformations: monomer-bridged, dimer-bridged and standing 765:837 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 3 28 theme μs 381:382 arg1 simulations 408:418 2.4 μs molecular dynamics (MD) simulations 377:418 2.4 μs molecular dynamics (MD) simulations 377:418 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 4 29 theme simulation 579:588 arg1 result 590:595 our simulation result 575:595 our simulation result 575:595 Moreover, with 146-N-glycan, our simulation result was more consistent with crystal structures of NAs than simulations conducted without glycans. 26267136 5 30 theme MD 716:717 arg1 trajectories 719:730 the MD trajectories 712:730 the MD trajectories 712:730 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 3 31 theme molecular 384:392 arg1 MD 404:405 MD 404:405 MD 404:405 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 3 31 theme molecular 384:392 arg1 dynamics 394:401 molecular dynamics 384:401 2.4 μs molecular dynamics (MD) simulations 377:418 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 0 32 theme Active 48:53 arg1 Neuraminidase 63:75 Neuraminidase 63:75 Neuraminidase 63:75 The Mechanism by which 146-N-Glycan Affects the Active Site of Neuraminidase. 26267136 0 32 theme Active 48:53 arg1 Site 55:58 the Active Site 44:58 the Active Site of Neuraminidase 44:75 The Mechanism by which 146-N-Glycan Affects the Active Site of Neuraminidase. 26267136 8 33 theme flexible 1230:1237 arg1 substituents 1239:1250 flexible substituents 1230:1250 flexible substituents 1230:1250 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 3 34 theme simulations 408:418 arg1 total 368:372 a total 366:372 a total of 2.4 μs molecular dynamics (MD) simulations 366:418 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 7 35 theme standing 1032:1039 arg1 conformations 1041:1053 various standing conformations 1024:1053 various standing conformations of 146-N-glycan 1024:1069 Furthermore, our simulation revealed that various standing conformations of 146-N-glycan could block the entrance of the binding pocket. 26267136 1 36 theme conserved 94:102 arg1 sites 118:122 the most conserved glycosylation sites 85:122 the most conserved glycosylation sites of neuraminidase (NA) 85:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 36 theme conserved 94:102 arg1 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 37 theme glycosylation 104:116 arg1 sites 118:122 the most conserved glycosylation sites 85:122 the most conserved glycosylation sites of neuraminidase (NA) 85:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 37 theme glycosylation 104:116 arg1 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 38 theme sites 118:122 arg1 sites 118:122 the most conserved glycosylation sites 85:122 the most conserved glycosylation sites of neuraminidase (NA) 85:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 38 theme sites 118:122 arg1 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 38 theme sites 118:122 arg1 One 78:80 One 78:80 One 78:80 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 9 39 theme better 1375:1380 arg1 inhibitors 1382:1391 better inhibitors 1375:1391 better inhibitors targeting the 150-cavity 1375:1416 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 9 39 theme better 1375:1380 arg1 substituents 1347:1358 rigid and hydrophobic substituents 1325:1358 rigid and hydrophobic substituents 1325:1358 Together, our results lead us to hypothesize that rigid and hydrophobic substituents could serve as better inhibitors targeting the 150-cavity. 26267136 4 40 theme NAs 644:646 arg1 structures 630:639 crystal structures 622:639 crystal structures of NAs 622:646 Moreover, with 146-N-glycan, our simulation result was more consistent with crystal structures of NAs than simulations conducted without glycans. 26267136 2 41 located found 219:223 arg1 site 243:246 the active site 232:246 the active site 232:246 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 2 41 located found 219:223 arg2 NA 206:207 NA 206:207 NA 206:207 This site is adjacent to the 150-cavity of NA, which is found within the active site and thought to be a target for rational drug development against the antiviral resistance of influenza. 26267136 5 42 theme monomer-bridged 795:809 arg1 conformations 780:792 three distinct conformations 765:792 three distinct conformations: monomer-bridged, dimer-bridged and standing 765:837 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 5 43 theme distinct 771:778 arg1 conformations 780:792 three distinct conformations 765:792 three distinct conformations: monomer-bridged, dimer-bridged and standing 765:837 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 1 44 theme neuraminidase 127:139 arg1 sites 118:122 the most conserved glycosylation sites 85:122 the most conserved glycosylation sites of neuraminidase (NA) 85:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 1 44 theme neuraminidase 127:139 arg1 neuraminidase 127:139 neuraminidase 127:139 neuraminidase (NA) 127:144 One of the most conserved glycosylation sites of neuraminidase (NA) is 146-N-glycan. 26267136 8 45 theme inhibitors 1214:1223 arg1 activity 1202:1209 the relatively low activity 1183:1209 the relatively low activity of inhibitors with flexible substituents toward the 150-cavity 1183:1272 This result was consistent with experimental data and explained the relatively low activity of inhibitors with flexible substituents toward the 150-cavity. 26267136 5 46 theme Cluster 692:698 arg1 analysis 700:707 Cluster analysis 692:707 Cluster analysis of the MD trajectories 692:730 Cluster analysis of the MD trajectories showed that 146-N-glycan adopted three distinct conformations: monomer-bridged, dimer-bridged and standing. 26267136 3 47 theme 150-cavity 534:543 arg1 volume 520:525 the volume 516:525 the volume of the 150-cavity 516:543 Here, through a total of 2.4 μs molecular dynamics (MD) simulations, we demonstrated that 146-N-glycan can stabilize the conformation of the 150-loop that controls the volume of the 150-cavity. 26267136 6 48 theme conformation 968:979 arg1 stabilization 938:950 stabilization 938:950 stabilization of the 150-loop conformation 938:979 Of these conformations, the dimer-bridged 146-N-glycan was the most stable one and contributed to stabilization of the 150-loop conformation. 24440233 4 0 theme structure 872:880 arg1 characterization 882:897 the putative glycan structure characterization 852:897 the putative glycan structure characterization 852:897 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 1 1 theme structure 142:150 arg1 elucidation 116:126 Simultaneous elucidation 103:126 Simultaneous elucidation of the glycan structure and the glycosylation site 103:177 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 10 2 theme structural 1821:1830 arg1 characterization 1832:1847 comprehensive structural characterization 1807:1847 comprehensive structural characterization of glycoproteins 1807:1864 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 6 3 from fragmented 1075:1084 arg1 contrast 1023:1030 contrast 1023:1030 contrast 1023:1030 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 4 4 theme putative 856:863 arg1 characterization 882:897 the putative glycan structure characterization 852:897 the putative glycan structure characterization 852:897 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 2 5 theme higher 315:320 arg1 HCD 355:357 higher energy collisional dissociation (HCD) 315:358 higher energy collisional dissociation (HCD) 315:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 10 6 theme single 1871:1876 arg1 analysis 1887:1894 a single LC-MS/MS analysis 1869:1894 a single LC-MS/MS analysis 1869:1894 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 5 7 theme unambiguous 951:961 arg1 differentiation 963:977 unambiguous differentiation 951:977 unambiguous differentiation of glycopeptides from non-glycopeptides 951:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 9 8 theme intact 1540:1545 arg1 glycopeptides 1547:1559 38 unique intact glycopeptides 1530:1559 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) 1530:1640 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 2 9 theme fragmentation 294:306 arg1 type 286:289 a recent type 277:289 a recent type of fragmentation termed higher energy collisional dissociation (HCD) 277:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 9 10 theme unique 1533:1538 arg1 glycopeptides 1547:1559 38 unique intact glycopeptides 1530:1559 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) 1530:1640 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 7 11 theme HCD 1201:1203 arg1 approach 1205:1212 the HCD approach 1197:1212 the HCD approach 1197:1212 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 1 12 theme protein 227:233 arg1 glycosylation 235:247 protein glycosylation 227:247 protein glycosylation 227:247 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 8 13 theme automated 1496:1504 arg1 analysis 1511:1518 automated data analysis 1496:1518 automated data analysis 1496:1518 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 4 14 attach attached 753:760 arg1 peptides 769:776 the peptides 765:776 the peptides 765:776 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 4 14 attach attached 753:760 arg2 chains 746:751 the sugar chains 736:751 the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization 736:897 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 3 15 theme optimal 574:580 arg1 conditions 582:591 the optimal conditions 570:591 the optimal conditions for the fragmentation of peptide backbones and glycoconjugates 570:654 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 1 16 theme glycan 135:140 arg1 structure 142:150 the glycan structure 131:150 the glycan structure 131:150 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 9 17 dep sites 1598:1602 arg1 N-linked 1609:1616 N-linked 1609:1616 N-linked 1609:1616 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 9 17 dep sites 1598:1602 arg1 sites 1635:1639 two O-linked sites 1622:1639 two O-linked sites 1622:1639 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 3 18 theme peptide 618:624 arg1 backbones 626:634 peptide backbones 618:634 peptide backbones 618:634 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 5 19 theme ions 938:941 arg1 detection 913:921 detection 913:921 detection of the oxonium ions 913:941 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 6 20 theme NCE 1049:1051 arg1 values 1053:1058 higher NCE values 1042:1058 higher NCE values 1042:1058 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 6 21 with HCD 1033:1035 arg1 values 1053:1058 higher NCE values 1042:1058 higher NCE values 1042:1058 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 7 22 theme N- 1287:1288 arg1 characterization 1260:1275 confident characterization 1250:1275 confident characterization of intact N- and O-linked glycopeptides 1250:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 1 23 theme glycosylation 160:172 arg1 site 174:177 the glycosylation site 156:177 the glycosylation site 156:177 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 4 24 theme NCE 699:701 arg1 values 703:708 lower NCE values 693:708 lower NCE values 693:708 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 0 25 theme collisional 77:87 arg1 dissociation 89:100 higher energy collisional dissociation 63:100 higher energy collisional dissociation 63:100 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 9 26 theme glycosylation 1584:1596 arg1 sites 1598:1602 eight glycosylation sites 1578:1602 eight glycosylation sites (six N-linked and two O-linked sites) 1578:1640 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 6 27 dep fragmented 1075:1084 arg1 provided 1118:1125 provided 1118:1125 provided information needed for confident peptide identification 1118:1181 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 7 28 theme confident 1250:1258 arg1 characterization 1260:1275 confident characterization 1250:1275 confident characterization of intact N- and O-linked glycopeptides 1250:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 6 29 from contrast 1023:1030 arg1 fragmented 1075:1084 fragmented 1075:1084 fragmented 1075:1084 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 2 30 theme glycopeptides 404:416 arg1 patterns 385:392 fragmentation patterns 371:392 fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins 371:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 2 31 theme proteins 468:475 arg1 mixture 435:441 a mixture 433:441 a mixture of standard glycosylated proteins 433:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 0 32 theme energy 70:75 arg1 dissociation 89:100 higher energy collisional dissociation 63:100 higher energy collisional dissociation 63:100 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 8 33 theme novel 1423:1427 arg1 pipeline 1443:1450 a novel data analysis pipeline 1421:1450 a novel data analysis pipeline 1421:1450 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 8 33 theme novel 1423:1427 arg1 GlycoFinder 1463:1473 so-called GlycoFinder 1453:1473 so-called GlycoFinder 1453:1473 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 1 34 theme Simultaneous 103:114 arg1 elucidation 116:126 Simultaneous elucidation 103:126 Simultaneous elucidation of the glycan structure and the glycosylation site 103:177 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 2 35 theme standard 446:453 arg1 proteins 468:475 standard glycosylated proteins 446:475 standard glycosylated proteins 446:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 8 36 theme analysis 1434:1441 arg1 pipeline 1443:1450 a novel data analysis pipeline 1421:1450 a novel data analysis pipeline 1421:1450 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 8 36 theme analysis 1434:1441 arg1 GlycoFinder 1463:1473 so-called GlycoFinder 1453:1473 so-called GlycoFinder 1453:1473 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 3 37 theme normalized 482:491 arg1 value 518:522 The normalized collisional energy (NCE) value 478:522 The normalized collisional energy (NCE) value for HCD 478:530 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 0 38 theme intact 20:25 arg1 N- 27:28 intact N- 20:28 intact N- 20:28 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 7 39 gly glycopeptides 1303:1315 arg2 glycopeptides 1303:1315 O-linked glycopeptides 1294:1315 O-linked glycopeptides 1294:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 10 40 theme concurrent 1726:1735 arg1 characterization 1737:1752 concurrent characterization 1726:1752 concurrent characterization of both the peptide and the glycan 1726:1787 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 7 41 theme spectrometry 1363:1374 arg1 analysis 1387:1394 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 0 42 theme higher 63:68 arg1 dissociation 89:100 higher energy collisional dissociation 63:100 higher energy collisional dissociation 63:100 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 3 43 theme NCE 513:515 arg1 value 518:522 The normalized collisional energy (NCE) value 478:522 The normalized collisional energy (NCE) value for HCD 478:530 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 0 44 theme glycopeptides 43:55 arg1 Characterization 0:15 Characterization 0:15 Characterization of intact N- and O-linked glycopeptides 0:55 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 6 45 theme peptide 1160:1166 arg1 identification 1168:1181 confident peptide identification 1150:1181 confident peptide identification 1150:1181 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 3 46 theme energy 505:510 arg1 value 518:522 The normalized collisional energy (NCE) value 478:522 The normalized collisional energy (NCE) value for HCD 478:530 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 1 47 theme glycosylation 235:247 arg1 function 215:222 the biological function 200:222 the biological function of protein glycosylation 200:247 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 7 48 theme single 1322:1327 arg1 analysis 1387:1394 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 4 49 theme neutral 802:808 arg1 loss 810:813 neutral loss 802:813 neutral loss of monosaccharides 802:832 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 2 50 theme fragmentation 371:383 arg1 patterns 385:392 fragmentation patterns 371:392 fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins 371:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 9 51 theme protein 1686:1692 arg1 mixture 1694:1700 a standard protein mixture 1675:1700 a standard protein mixture 1675:1700 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 10 52 theme glycan 1782:1787 arg1 characterization 1737:1752 concurrent characterization 1726:1752 concurrent characterization of both the peptide and the glycan 1726:1787 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 4 53 theme glycan 865:870 arg1 characterization 882:897 the putative glycan structure characterization 852:897 the putative glycan structure characterization 852:897 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 7 54 theme O-linked 1294:1301 arg1 glycopeptides 1303:1315 O-linked glycopeptides 1294:1315 O-linked glycopeptides 1294:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 2 55 theme recent 279:284 arg1 type 286:289 a recent type 277:289 a recent type of fragmentation termed higher energy collisional dissociation (HCD) 277:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 4 56 with HCD 684:686 arg1 values 703:708 lower NCE values 693:708 lower NCE values 693:708 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 2 57 theme energy 322:327 arg1 HCD 355:357 higher energy collisional dissociation (HCD) 315:358 higher energy collisional dissociation (HCD) 315:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 10 58 theme comprehensive 1807:1819 arg1 characterization 1832:1847 comprehensive structural characterization 1807:1847 comprehensive structural characterization of glycoproteins 1807:1864 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 0 59 link O-linked 34:41 arg1 glycopeptides 43:55 O-linked glycopeptides 34:55 O-linked glycopeptides 34:55 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 8 60 theme so-called 1453:1461 arg1 pipeline 1443:1450 a novel data analysis pipeline 1421:1450 a novel data analysis pipeline 1421:1450 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 8 60 theme so-called 1453:1461 arg1 GlycoFinder 1463:1473 so-called GlycoFinder 1453:1473 so-called GlycoFinder 1453:1473 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 10 61 theme glycoproteins 1852:1864 arg1 characterization 1832:1847 comprehensive structural characterization 1807:1847 comprehensive structural characterization of glycoproteins 1807:1864 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 1 62 theme biological 204:213 arg1 function 215:222 the biological function 200:222 the biological function of protein glycosylation 200:247 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 7 63 link O-linked 1294:1301 arg1 glycopeptides 1303:1315 O-linked glycopeptides 1294:1315 O-linked glycopeptides 1294:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 7 64 theme chromatography-tandem 1336:1356 arg1 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 7 64 theme chromatography-tandem 1336:1356 arg1 spectrometry 1363:1374 liquid chromatography-tandem mass spectrometry 1329:1374 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 8 65 theme data 1506:1509 arg1 analysis 1511:1518 automated data analysis 1496:1518 automated data analysis 1496:1518 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 1 66 gly glycosylation 160:172 arg2 site 174:177 the glycosylation site 156:177 the glycosylation site 156:177 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 10 67 theme LC-MS/MS 1878:1885 arg1 analysis 1887:1894 a single LC-MS/MS analysis 1869:1894 a single LC-MS/MS analysis 1869:1894 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 5 68 theme oxonium 930:936 arg1 ions 938:941 the oxonium ions 926:941 the oxonium ions 926:941 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 2 69 theme dissociation 341:352 arg1 HCD 355:357 higher energy collisional dissociation (HCD) 315:358 higher energy collisional dissociation (HCD) 315:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 7 70 theme intact 1280:1285 arg1 N- 1287:1288 intact N- 1280:1288 intact N- 1280:1288 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 3 71 dep 60 554:555 arg1 to 551:552 to 551:552 to 551:552 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 6 72 theme higher 1042:1047 arg1 values 1053:1058 higher NCE values 1042:1058 higher NCE values 1042:1058 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 5 73 gly glycopeptides 982:994 arg1 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 5 73 gly glycopeptides 982:994 arg2 glycopeptides 982:994 glycopeptides 982:994 glycopeptides from non-glycopeptides 982:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 1 74 theme site 174:177 arg1 elucidation 116:126 Simultaneous elucidation 103:126 Simultaneous elucidation of the glycan structure and the glycosylation site 103:177 Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. 24440233 5 75 gly non-glycopeptides 1001:1017 arg2 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 9 76 gly glycopeptides 1547:1559 arg2 glycopeptides 1547:1559 38 unique intact glycopeptides 1530:1559 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) 1530:1640 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 4 77 theme lower 693:697 arg1 values 703:708 lower NCE values 693:708 lower NCE values 693:708 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 5 78 theme glycopeptides 982:994 arg1 differentiation 963:977 unambiguous differentiation 951:977 unambiguous differentiation of glycopeptides from non-glycopeptides 951:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 7 79 theme NCE 1231:1233 arg1 parameters 1235:1244 NCE parameters 1231:1244 NCE parameters for confident characterization of intact N- and O-linked glycopeptides 1231:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 9 80 gly glycosylation 1584:1596 arg2 sites 1598:1602 eight glycosylation sites 1578:1602 eight glycosylation sites (six N-linked and two O-linked sites) 1578:1640 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 9 80 gly glycosylation 1584:1596 arg2 eight 1578:1582 eight 1578:1582 eight 1578:1582 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 2 81 gly glycopeptides 404:416 arg2 glycopeptides 404:416 intact glycopeptides 397:416 intact glycopeptides generated from a mixture of standard glycosylated proteins 397:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 2 82 theme intact 397:402 arg1 glycopeptides 404:416 intact glycopeptides 397:416 intact glycopeptides generated from a mixture of standard glycosylated proteins 397:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 2 83 theme glycosylated 455:466 arg1 proteins 468:475 standard glycosylated proteins 446:475 standard glycosylated proteins 446:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 4 84 theme sugar 740:744 arg1 chains 746:751 the sugar chains 736:751 the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization 736:897 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 0 85 theme N- 27:28 arg1 Characterization 0:15 Characterization 0:15 Characterization of intact N- and O-linked glycopeptides 0:55 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 9 86 theme standard 1677:1684 arg1 mixture 1694:1700 a standard protein mixture 1675:1700 a standard protein mixture 1675:1700 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 9 87 theme O-linked 1626:1633 arg1 sites 1635:1639 two O-linked sites 1622:1639 two O-linked sites 1622:1639 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 6 88 theme peptide 1090:1096 arg1 backbone 1098:1105 the peptide backbone 1086:1105 the peptide backbone 1086:1105 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 3 89 theme glycoconjugates 640:654 arg1 fragmentation 601:613 the fragmentation 597:613 the fragmentation of peptide backbones and glycoconjugates 597:654 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 8 90 theme data 1429:1432 arg1 pipeline 1443:1450 a novel data analysis pipeline 1421:1450 a novel data analysis pipeline 1421:1450 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 8 90 theme data 1429:1432 arg1 GlycoFinder 1463:1473 so-called GlycoFinder 1453:1473 so-called GlycoFinder 1453:1473 In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. 24440233 3 91 theme collisional 493:503 arg1 value 518:522 The normalized collisional energy (NCE) value 478:522 The normalized collisional energy (NCE) value for HCD 478:530 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 0 92 theme O-linked 34:41 arg1 glycopeptides 43:55 O-linked glycopeptides 34:55 O-linked glycopeptides 34:55 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 10 93 gly glycoproteins 1852:1864 arg1 glycoproteins 1852:1864 glycoproteins 1852:1864 glycoproteins 1852:1864 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 7 94 theme mass 1358:1361 arg1 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 7 94 theme mass 1358:1361 arg1 spectrometry 1363:1374 liquid chromatography-tandem mass spectrometry 1329:1374 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 3 95 theme backbones 626:634 arg1 fragmentation 601:613 the fragmentation 597:613 the fragmentation of peptide backbones and glycoconjugates 597:654 The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. 24440233 4 96 theme loss 810:813 arg1 ladder 792:797 a ladder 790:797 a ladder of neutral loss of monosaccharides 790:832 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 7 97 theme liquid 1329:1334 arg1 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 LC-MS/MS 1377:1384 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 7 97 theme liquid 1329:1334 arg1 spectrometry 1363:1374 liquid chromatography-tandem mass spectrometry 1329:1374 a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 1320:1394 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 4 98 theme monosaccharides 818:832 arg1 loss 810:813 neutral loss 802:813 neutral loss of monosaccharides 802:832 Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. 24440233 10 99 theme peptide 1766:1772 arg1 characterization 1737:1752 concurrent characterization 1726:1752 concurrent characterization of both the peptide and the glycan 1726:1787 This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis. 24440233 5 100 from differentiation 963:977 arg1 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 non-glycopeptides 1001:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 2 101 gly glycosylated 455:466 arg1 proteins 468:475 standard glycosylated proteins 446:475 standard glycosylated proteins 446:475 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 6 102 theme confident 1150:1158 arg1 identification 1168:1181 confident peptide identification 1150:1181 confident peptide identification 1150:1181 In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. 24440233 2 103 theme collisional 329:339 arg1 HCD 355:357 higher energy collisional dissociation (HCD) 315:358 higher energy collisional dissociation (HCD) 315:358 In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. 24440233 9 104 link O-linked 1626:1633 arg1 sites 1635:1639 two O-linked sites 1622:1639 two O-linked sites 1622:1639 Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. 24440233 0 105 gly glycopeptides 43:55 arg2 glycopeptides 43:55 O-linked glycopeptides 34:55 O-linked glycopeptides 34:55 Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation. 24440233 7 106 theme glycopeptides 1303:1315 arg1 characterization 1260:1275 confident characterization 1250:1275 confident characterization of intact N- and O-linked glycopeptides 1250:1315 We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. 24440233 5 107 from non-glycopeptides 1001:1017 arg1 differentiation 963:977 unambiguous differentiation 951:977 unambiguous differentiation of glycopeptides from non-glycopeptides 951:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 24440233 5 107 from non-glycopeptides 1001:1017 arg1 glycopeptides 982:994 glycopeptides 982:994 glycopeptides from non-glycopeptides 982:1017 In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. 27932382 10 0 theme development 1869:1879 arg1 stage 1849:1853 the phylotypic stage 1834:1853 the phylotypic stage of vertebrate development 1834:1879 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 1 1 theme vertebrate 128:137 arg1 development 139:149 vertebrate development 128:149 vertebrate development 128:149 Zebrafish is a model organism suitable for studying vertebrate development. 27932382 4 2 with glycans 619:625 arg1 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 2 with glycans 619:625 arg1 LacNAc 632:637 LacNAc 632:637 LacNAc (Galβ1-4GlcNAc) 632:653 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 2 with glycans 619:625 arg1 LacNAc 669:674 sialyl LacNAc 662:674 sialyl LacNAc 662:674 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 8 3 from stages 1310:1315 arg1 components 1362:1371 minor components 1356:1371 minor components 1356:1371 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 8 3 from stages 1310:1315 arg1 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 8 3 from stages 1310:1315 arg1 LacdiNAc 1422:1429 sialyl LacdiNAc 1415:1429 sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc) 1415:1455 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 6 4 with glycans 1049:1055 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 7 5 theme deyolked 1168:1175 arg1 embryos 1177:1183 deyolked embryos 1168:1183 deyolked embryos 1168:1183 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 9 6 theme antenna 1550:1556 arg1 structures 1558:1567 LacNAc antenna structures 1543:1567 LacNAc antenna structures 1543:1567 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 4 7 from postfertilization 512:528 arg1 embryos 492:498 deyolked embryos 483:498 deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, 483:553 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 6 8 theme oligomannose-type 1031:1047 arg1 glycans 1049:1055 predominant oligomannose-type glycans 1019:1055 predominant oligomannose-type glycans 1019:1055 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 8 9 theme minor 1356:1360 arg1 components 1362:1371 minor components 1356:1371 minor components 1356:1371 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 7 10 theme N-Glycan 1136:1143 arg1 profiles 1145:1152 N-Glycan profiles 1136:1152 N-Glycan profiles obtained from deyolked embryos at different stages 1136:1203 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 2 11 theme N-glycosylation 309:323 arg1 roles 300:304 the roles 296:304 the roles of N-glycosylation 296:323 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 8 12 theme segmentation 1297:1308 arg1 stages 1310:1315 gastrula and early segmentation stages 1278:1315 stages 1310:1315 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 4 13 from embryos 492:498 arg1 N-glycans 468:476 The N-glycans 464:476 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, 464:553 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 3 14 theme Multiple 326:333 arg1 modes 335:339 Multiple modes 326:339 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry 326:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 9 15 dep stages 1498:1503 arg1 the 1463:1465 the 1463:1465 the 1463:1465 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 9 15 dep stages 1498:1503 arg1 mid-segmentation 1467:1482 mid-segmentation 1467:1482 mid-segmentation 1467:1482 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 10 16 theme mechanisms 1742:1751 arg1 presence 1730:1737 the presence 1726:1737 the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development 1726:1879 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 9 17 theme α2,3-sialyl 1606:1616 arg1 antenna 1625:1631 α2,3-sialyl LacNAc antenna 1606:1631 α2,3-sialyl LacNAc antenna 1606:1631 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 4 18 theme oligomannose-type 772:788 arg1 glycans 790:796 abundant oligomannose-type glycans 763:796 abundant oligomannose-type glycans 763:796 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 6 19 theme Siaα2-3 1095:1101 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 3 20 theme liquid 361:366 arg1 chromatography 368:381 high-performance liquid chromatography 344:381 high-performance liquid chromatography 344:381 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 9 21 dep LacNAc 1517:1522 arg1 structures 1558:1567 LacNAc antenna structures 1543:1567 LacNAc antenna structures 1543:1567 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 0 22 theme embryos 67:73 arg1 N-glycans 44:52 N-glycans 44:52 N-glycans of zebrafish embryos 44:73 Structures and developmental alterations of N-glycans of zebrafish embryos. 27932382 9 23 theme core 1634:1637 arg1 α1,6-Fuc 1639:1646 core α1,6-Fuc 1634:1646 core α1,6-Fuc 1634:1646 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 6 24 theme Galβ1-4 1087:1093 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 5 25 theme hybrid-type 824:834 arg1 glycans 836:842 the complex- and hybrid-type glycans 807:842 the complex- and hybrid-type glycans 807:842 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 4 26 theme deyolked 483:490 arg1 embryos 492:498 deyolked embryos 483:498 deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, 483:553 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 3 27 theme mass 398:401 arg1 spectrometry 403:414 multistage mass spectrometry 387:414 multistage mass spectrometry 387:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 4 28 theme abundant 763:770 arg1 glycans 790:796 abundant oligomannose-type glycans 763:796 abundant oligomannose-type glycans 763:796 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 10 29 theme complex/hybrid 1794:1807 arg1 biosynthesis 1818:1829 complex/hybrid N-glycan biosynthesis 1794:1829 complex/hybrid N-glycan biosynthesis 1794:1829 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 7 30 theme glycans 1266:1272 arg1 variation 1228:1236 stage-dependent variation 1212:1236 stage-dependent variation of complex- and hybrid-type glycans 1212:1272 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 1 31 theme model 91:95 arg1 Zebrafish 76:84 Zebrafish 76:84 Zebrafish 76:84 Zebrafish is a model organism suitable for studying vertebrate development. 27932382 1 31 theme model 91:95 arg1 organism 97:104 a model organism 89:104 a model organism suitable for studying vertebrate development 89:149 Zebrafish is a model organism suitable for studying vertebrate development. 27932382 6 32 theme GlcNAc 1119:1124 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 0 33 theme developmental 15:27 arg1 alterations 29:39 developmental alterations 15:39 developmental alterations 15:39 Structures and developmental alterations of N-glycans of zebrafish embryos. 27932382 1 34 theme suitable 106:113 arg1 Zebrafish 76:84 Zebrafish 76:84 Zebrafish 76:84 Zebrafish is a model organism suitable for studying vertebrate development. 27932382 1 34 theme suitable 106:113 arg1 organism 97:104 a model organism 89:104 a model organism suitable for studying vertebrate development 89:149 Zebrafish is a model organism suitable for studying vertebrate development. 27932382 6 35 theme Fucα1-3 1111:1117 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 3 36 theme N-glycans 453:461 arg1 analysis 441:448 structural analysis 430:448 structural analysis of N-glycans 430:461 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 6 37 contain contains 1010:1017 arg2 glycans 1049:1055 predominant oligomannose-type glycans 1019:1055 predominant oligomannose-type glycans 1019:1055 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 6 37 contain contains 1010:1017 arg1 contrast 977:984 contrast 977:984 contrast 977:984 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 6 37 contain contains 1010:1017 arg1 yolk 991:994 the yolk 987:994 the yolk of the embryo 987:1008 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 6 37 contain contains 1010:1017 arg2 yolk 991:994 the yolk 987:994 the yolk of the embryo 987:1008 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 6 37 contain contains 1010:1017 arg2 glycans 1074:1080 complex-type glycans 1061:1080 complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1061:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 10 38 theme biosynthesis 1818:1829 arg1 structures 1780:1789 the antenna structures 1768:1789 the antenna structures of complex/hybrid N-glycan biosynthesis 1768:1829 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 6 39 theme Galβ1-4 1103:1109 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 4 40 theme higher 576:581 arg1 complex- 594:601 complex- 594:601 complex- 594:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 40 theme higher 576:581 arg1 amounts 583:589 relatively higher amounts 565:589 relatively higher amounts of complex- 565:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 2 41 theme N-glycan 168:175 arg1 structures 177:186 the N-glycan structures 164:186 the N-glycan structures of zebrafish embryos 164:207 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 10 42 theme vertebrate 1858:1867 arg1 development 1869:1879 vertebrate development 1858:1879 vertebrate development 1858:1879 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 5 43 theme extended 862:869 arg1 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 43 theme extended 862:869 arg1 structures 878:887 various extended LacNAc structures 854:887 various extended LacNAc structures 854:887 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 43 theme extended 862:869 arg1 LacNAc-repeat 913:925 LacNAc-repeat 913:925 LacNAc-repeat 913:925 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 43 theme extended 862:869 arg1 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 44 theme complex- 811:818 arg1 glycans 836:842 the complex- and hybrid-type glycans 807:842 the complex- and hybrid-type glycans 807:842 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 9 45 theme pharyngula 1487:1496 arg1 stages 1498:1503 pharyngula stages 1487:1503 pharyngula stages 1487:1503 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 2 46 theme embryos 201:207 arg1 structures 177:186 the N-glycan structures 164:186 the N-glycan structures of zebrafish embryos 164:207 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 2 46 theme embryos 201:207 arg1 alterations 219:229 their alterations 213:229 their alterations during zebrafish embryogenesis 213:260 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 4 47 contain contained 555:563 arg2 glycans 619:625 hybrid-type glycans 607:625 hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal 607:702 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 47 contain contained 555:563 arg2 complex- 594:601 complex- 594:601 complex- 594:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 47 contain contained 555:563 arg1 N-glycans 468:476 The N-glycans 464:476 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, 464:553 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 47 contain contained 555:563 arg2 amounts 583:589 relatively higher amounts 565:589 relatively higher amounts of complex- 565:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 5 48 theme unique 930:935 arg1 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 0 49 theme zebrafish 57:65 arg1 embryos 67:73 zebrafish embryos 57:73 zebrafish embryos 57:73 Structures and developmental alterations of N-glycans of zebrafish embryos. 27932382 7 50 theme different 1188:1196 arg1 stages 1198:1203 different stages 1188:1203 different stages 1188:1203 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 9 51 theme LacNAc 1543:1548 arg1 structures 1558:1567 LacNAc antenna structures 1543:1567 LacNAc antenna structures 1543:1567 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 7 52 theme stage-dependent 1212:1226 arg1 variation 1228:1236 stage-dependent variation 1212:1236 stage-dependent variation of complex- and hybrid-type glycans 1212:1272 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 8 53 theme complex- 1318:1325 arg1 glycans 1343:1349 complex- and hybrid-type glycans 1318:1349 complex- and hybrid-type glycans 1318:1349 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 8 54 theme hybrid-type 1331:1341 arg1 glycans 1343:1349 complex- and hybrid-type glycans 1318:1349 complex- and hybrid-type glycans 1318:1349 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 4 55 with amounts 583:589 arg1 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 55 with amounts 583:589 arg1 LacNAc 632:637 LacNAc 632:637 LacNAc (Galβ1-4GlcNAc) 632:653 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 55 with amounts 583:589 arg1 LacNAc 669:674 sialyl LacNAc 662:674 sialyl LacNAc 662:674 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 8 56 theme gastrula 1278:1285 arg1 stages 1310:1315 gastrula and early segmentation stages 1278:1315 stages 1310:1315 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 6 57 theme predominant 1019:1029 arg1 glycans 1049:1055 predominant oligomannose-type glycans 1019:1055 predominant oligomannose-type glycans 1019:1055 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 4 58 theme additional 684:693 arg1 β1,4-Gal 695:702 additional β1,4-Gal 684:702 additional β1,4-Gal 684:702 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 8 59 theme early 1291:1295 arg1 stages 1310:1315 gastrula and early segmentation stages 1278:1315 stages 1310:1315 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 6 60 theme embryo 1003:1008 arg1 yolk 991:994 the yolk 987:994 the yolk of the embryo 987:1008 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 4 61 located found 724:728 arg2 glycans 790:796 abundant oligomannose-type glycans 763:796 abundant oligomannose-type glycans 763:796 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 61 located found 724:728 arg2 glycans 619:625 hybrid-type glycans 607:625 hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal 607:702 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 61 located found 724:728 arg1 tissues 743:749 mammalian tissues 733:749 mammalian tissues 733:749 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 2 62 theme zebrafish 238:246 arg1 embryogenesis 248:260 zebrafish embryogenesis 238:260 zebrafish embryogenesis 238:260 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 4 63 with glycans 790:796 arg1 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 Galβ1-4GlcNAc 640:652 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 63 with glycans 790:796 arg1 LacNAc 632:637 LacNAc 632:637 LacNAc (Galβ1-4GlcNAc) 632:653 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 63 with glycans 790:796 arg1 LacNAc 669:674 sialyl LacNAc 662:674 sialyl LacNAc 662:674 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 9 64 theme LacNAc 1618:1623 arg1 antenna 1625:1631 α2,3-sialyl LacNAc antenna 1606:1631 α2,3-sialyl LacNAc antenna 1606:1631 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 5 65 contain possessed 844:852 arg2 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 65 contain possessed 844:852 arg2 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 65 contain possessed 844:852 arg1 Some 799:802 Some 799:802 Some 799:802 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 65 contain possessed 844:852 arg2 structures 878:887 various extended LacNAc structures 854:887 various extended LacNAc structures 854:887 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 65 contain possessed 844:852 arg2 LacNAc-repeat 913:925 LacNAc-repeat 913:925 LacNAc-repeat 913:925 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 65 contain possessed 844:852 arg1 glycans 836:842 the complex- and hybrid-type glycans 807:842 the complex- and hybrid-type glycans 807:842 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 8 66 theme antenna 1384:1390 arg1 structures 1392:1401 their antenna structures 1378:1401 their antenna structures 1378:1401 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 2 67 theme basic 272:276 arg1 data 278:281 basic data 272:281 basic data for studying the roles of N-glycosylation 272:323 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 5 68 theme dHex 942:945 arg1 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 3 69 theme chromatography 368:381 arg1 modes 335:339 Multiple modes 326:339 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry 326:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 6 70 with glycans 1074:1080 arg1 antennae 1126:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1087:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 4 71 theme mammalian 733:741 arg1 tissues 743:749 mammalian tissues 733:749 mammalian tissues 733:749 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 3 72 theme high-performance 344:359 arg1 chromatography 368:381 high-performance liquid chromatography 344:381 high-performance liquid chromatography 344:381 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 8 73 theme sialyl 1415:1420 arg1 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 Siaα2-6GalNAcβ1-4GlcNAc 1432:1454 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 8 73 theme sialyl 1415:1420 arg1 LacdiNAc 1422:1429 sialyl LacdiNAc 1415:1429 sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc) 1415:1455 At gastrula and early segmentation stages, complex- and hybrid-type glycans were minor components, and their antenna structures were mainly sialyl LacdiNAc (Siaα2-6GalNAcβ1-4GlcNAc). 27932382 9 74 theme bisecting 1652:1660 arg1 GlcNAc 1662:1667 bisecting GlcNAc 1652:1667 bisecting GlcNAc 1652:1667 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 6 75 theme complex-type 1061:1072 arg1 glycans 1074:1080 complex-type glycans 1061:1080 complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae 1061:1133 In contrast, the yolk of the embryo contains predominant oligomannose-type glycans and complex-type glycans with Galβ1-4(Siaα2-3)Galβ1-4(Fucα1-3)GlcNAc antennae. 27932382 3 76 theme spectrometry 403:414 arg1 modes 335:339 Multiple modes 326:339 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry 326:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 9 77 dep antenna 1625:1631 arg1 modifications 1669:1681 modifications 1669:1681 modifications 1669:1681 From the mid-segmentation to pharyngula stages, those with LacNAc and/or α2,6-sialyl LacNAc antenna structures increased remarkably, and those with α2,3-sialyl LacNAc antenna, core α1,6-Fuc and bisecting GlcNAc modifications increased gradually. 27932382 3 78 theme multistage 387:396 arg1 spectrometry 403:414 multistage mass spectrometry 387:414 multistage mass spectrometry 387:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 4 79 theme mid-pharyngula 533:546 arg1 postfertilization 512:528 36 hours postfertilization 503:528 36 hours postfertilization 503:528 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 79 theme mid-pharyngula 533:546 arg1 stage 548:552 a mid-pharyngula stage 531:552 a mid-pharyngula stage 531:552 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 5 80 theme LacNAc 871:876 arg1 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 80 theme LacNAc 871:876 arg1 structures 878:887 various extended LacNAc structures 854:887 various extended LacNAc structures 854:887 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 80 theme LacNAc 871:876 arg1 LacNAc-repeat 913:925 LacNAc-repeat 913:925 LacNAc-repeat 913:925 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 80 theme LacNAc 871:876 arg1 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 3 81 theme structural 430:439 arg1 analysis 441:448 structural analysis 430:448 structural analysis of N-glycans 430:461 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 4 82 theme hybrid-type 607:617 arg1 glycans 619:625 hybrid-type glycans 607:625 hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal 607:702 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 10 83 theme antenna 1772:1778 arg1 structures 1780:1789 the antenna structures 1768:1789 the antenna structures of complex/hybrid N-glycan biosynthesis 1768:1829 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 7 84 theme complex- 1241:1248 arg1 glycans 1266:1272 complex- and hybrid-type glycans 1241:1272 complex- and hybrid-type glycans 1241:1272 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27932382 3 85 used used 421:424 arg2 modes 335:339 Multiple modes 326:339 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry 326:414 Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. 27932382 2 86 theme zebrafish 191:199 arg1 embryos 201:207 zebrafish embryos 191:207 zebrafish embryos 191:207 We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. 27932382 0 87 theme N-glycans 44:52 arg1 Structures 0:9 Structures 0:9 Structures 0:9 Structures and developmental alterations of N-glycans of zebrafish embryos. 27932382 0 87 theme N-glycans 44:52 arg1 alterations 29:39 developmental alterations 15:39 developmental alterations 15:39 Structures and developmental alterations of N-glycans of zebrafish embryos. 27932382 10 88 theme N-glycan 1809:1816 arg1 biosynthesis 1818:1829 complex/hybrid N-glycan biosynthesis 1794:1829 complex/hybrid N-glycan biosynthesis 1794:1829 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 5 89 theme various 854:860 arg1 -GalNAcα1-GlcNAcβ1-LacNAc 947:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc 930:971 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 89 theme various 854:860 arg1 structures 878:887 various extended LacNAc structures 854:887 various extended LacNAc structures 854:887 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 89 theme various 854:860 arg1 LacNAc-repeat 913:925 LacNAc-repeat 913:925 LacNAc-repeat 913:925 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 5 89 theme various 854:860 arg1 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Galβ1-4LacNAc 898:910 Some of the complex- and hybrid-type glycans possessed various extended LacNAc structures, such as Galβ1-4LacNAc, LacNAc-repeat or unique (+/- dHex)-GalNAcα1-GlcNAcβ1-LacNAc. 27932382 4 90 theme complex- 594:601 arg1 glycans 790:796 abundant oligomannose-type glycans 763:796 abundant oligomannose-type glycans 763:796 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 90 theme complex- 594:601 arg1 complex- 594:601 complex- 594:601 complex- 594:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 90 theme complex- 594:601 arg1 glycans 619:625 hybrid-type glycans 607:625 hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal 607:702 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 90 theme complex- 594:601 arg1 amounts 583:589 relatively higher amounts 565:589 relatively higher amounts of complex- 565:601 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 91 theme hours 506:510 arg1 postfertilization 512:528 36 hours postfertilization 503:528 36 hours postfertilization 503:528 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 91 theme hours 506:510 arg1 stage 548:552 a mid-pharyngula stage 531:552 a mid-pharyngula stage 531:552 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 4 92 theme sialyl 662:667 arg1 LacNAc 669:674 sialyl LacNAc 662:674 sialyl LacNAc 662:674 The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans. 27932382 10 93 theme phylotypic 1838:1847 arg1 stage 1849:1853 the phylotypic stage 1834:1853 the phylotypic stage of vertebrate development 1834:1879 These results suggest the presence of mechanisms for regulating the antenna structures of complex/hybrid N-glycan biosynthesis in the phylotypic stage of vertebrate development. 27932382 7 94 theme hybrid-type 1254:1264 arg1 glycans 1266:1272 complex- and hybrid-type glycans 1241:1272 complex- and hybrid-type glycans 1241:1272 N-Glycan profiles obtained from deyolked embryos at different stages showed stage-dependent variation of complex- and hybrid-type glycans. 27743372 6 0 theme bead 991:994 arg1 preparation 1022:1032 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 10 1 theme IgG 1631:1633 arg1 glycans 1635:1641 APTS labeled IgG glycans 1618:1641 APTS labeled IgG glycans 1618:1641 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 3 2 theme analysis 582:589 arg1 importance 562:571 the importance 558:571 the importance of their analysis 558:589 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 4 3 theme glycan 662:667 arg1 alterations 679:689 disease-related glycan structure alterations 646:689 disease-related glycan structure alterations 646:689 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 7 4 theme induced 1199:1205 arg1 CE-LIF 1231:1236 CE-LIF 1231:1236 CE-LIF 1231:1236 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 7 4 theme induced 1199:1205 arg1 detection 1220:1228 laser induced fluorescence detection 1193:1228 laser induced fluorescence detection (CE-LIF) 1193:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 6 5 gly glycoprotein 1002:1013 arg1 glycoprotein 1002:1013 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 6 6 theme field 969:973 arg1 developments 949:960 the newest exciting developments 929:960 the newest exciting developments of the field 929:973 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 4 7 theme new 729:731 arg1 biomarkers 733:742 new biomarkers 729:742 new biomarkers for early diagnostics 729:764 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 0 8 theme High-Throughput 0:14 arg1 Analysis 25:32 High-Throughput N-Glycan Analysis 0:32 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation 0:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 1 9 theme promising 191:199 arg1 drugs 216:220 these promising new-generation drugs 185:220 these promising new-generation drugs 185:220 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 8 10 theme endoglycosidase 1270:1284 arg1 F 1293:1293 the endoglycosidase PNGase F 1266:1293 the endoglycosidase PNGase F 1266:1293 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 7 11 theme prepared 1173:1180 arg1 samples 1182:1188 the prepared samples 1169:1188 the prepared samples 1169:1188 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 9 12 theme plate 1553:1557 arg1 formats 1559:1565 individual (single vial) and 96-well plate formats 1516:1565 formats 1559:1565 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 8 13 theme fluorophore 1355:1365 arg1 dye 1367:1369 the charged fluorophore dye 1343:1369 the charged fluorophore dye of aminopyrenetrisulfonate (APTS) 1343:1403 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 10 14 theme CE-LIF 1680:1685 arg1 protocol 1687:1694 an optimized CE-LIF protocol 1667:1694 an optimized CE-LIF protocol 1667:1694 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 5 15 dep applicable 813:822 arg1 high-throughput 825:839 high-throughput 825:839 high-throughput 825:839 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 4 16 theme discovery 716:724 arg1 promise 701:707 the promise 697:707 the promise of the discovery of new biomarkers for early diagnostics 697:764 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 7 17 theme electrophoresis 1141:1155 arg1 analysis 1157:1164 rapid capillary electrophoresis analysis 1125:1164 rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF) 1125:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 9 18 theme excess 1578:1583 arg1 removal 1589:1595 excess dye removal 1578:1595 excess dye removal 1578:1595 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 6 19 theme preparation 1022:1032 arg1 technique 1034:1042 the magnetic bead based glycoprotein sample preparation technique 978:1042 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 7 20 theme rapid 1125:1129 arg1 electrophoresis 1141:1155 rapid capillary electrophoresis 1125:1155 rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF) 1125:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 4 21 theme biomarkers 733:742 arg1 discovery 716:724 the discovery 712:724 the discovery of new biomarkers for early diagnostics 712:764 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 6 22 theme glycoprotein 1002:1013 arg1 preparation 1022:1032 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 7 23 theme method 1073:1078 arg1 protocol 1056:1063 A detailed protocol 1045:1063 A detailed protocol of this method 1045:1078 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 5 24 theme sample 841:846 arg1 preparation 848:858 sample preparation 841:858 sample preparation 841:858 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 7 25 theme detailed 1047:1054 arg1 protocol 1056:1063 A detailed protocol 1045:1063 A detailed protocol of this method 1045:1078 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 3 26 theme multibillion 498:509 arg1 business 528:535 multibillion dollar worldwide business 498:535 multibillion dollar worldwide business 498:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 1 27 gly glycoproteins 119:131 arg1 glycoproteins 119:131 therapeutic glycoproteins 107:131 therapeutic glycoproteins 107:131 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 9 28 theme glycan 1416:1421 arg1 capture 1423:1429 Effective glycan capture 1406:1429 Effective glycan capture by magnetic microparticles 1406:1456 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 0 29 theme Magnetic 45:52 arg1 Preparation 72:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 7 30 with conjunction 1108:1118 arg1 analysis 1157:1164 rapid capillary electrophoresis analysis 1125:1164 rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF) 1125:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 3 31 theme than 431:434 arg1 glycoproteins 471:483 glycoproteins 471:483 glycoproteins representing multibillion dollar worldwide business 471:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 3 31 theme than 431:434 arg1 half 436:439 more than half 426:439 more than half of the biopharmaceuticals 426:465 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 5 32 theme N-glycan 885:892 arg1 profiling 894:902 N-glycan profiling 885:902 N-glycan profiling of glycoproteins 885:919 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 1 33 theme therapeutic 107:117 arg1 glycoproteins 119:131 therapeutic glycoproteins 107:131 therapeutic glycoproteins 107:131 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 10 34 theme Rapid 1598:1602 arg1 separation 1604:1613 Rapid separation 1598:1613 Rapid separation of APTS labeled IgG glycans 1598:1641 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 1 35 dep activity 160:167 arg1 the 156:158 the 156:158 the 156:158 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 6 36 theme developments 949:960 arg1 One 922:924 One 922:924 One 922:924 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 6 36 theme developments 949:960 arg1 developments 949:960 the newest exciting developments 929:960 the newest exciting developments of the field 929:973 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 3 37 theme biopharmaceuticals 448:465 arg1 glycoproteins 471:483 glycoproteins 471:483 glycoproteins representing multibillion dollar worldwide business 471:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 3 37 theme biopharmaceuticals 448:465 arg1 half 436:439 more than half 426:439 more than half of the biopharmaceuticals 426:465 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 4 38 theme other 624:628 arg1 hand 630:633 the other hand 620:633 the other hand 620:633 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 3 39 theme worldwide 518:526 arg1 business 528:535 multibillion dollar worldwide business 498:535 multibillion dollar worldwide business 498:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 0 40 theme Rapid 39:43 arg1 Preparation 72:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 9 41 theme fast 1466:1469 arg1 preparation 1496:1506 fast, easily automated sample preparation 1466:1506 fast, easily automated sample preparation 1466:1506 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 6 42 theme newest 933:938 arg1 developments 949:960 the newest exciting developments 929:960 the newest exciting developments of the field 929:973 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 10 43 theme labeled 1623:1629 arg1 glycans 1635:1641 APTS labeled IgG glycans 1618:1641 APTS labeled IgG glycans 1618:1641 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 0 44 with Analysis 25:32 arg1 Preparation 72:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 2 45 theme stability 391:399 arg1 properties 401:410 physicochemical and thermal stability properties 363:410 physicochemical and thermal stability properties 363:410 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 0 46 theme Bead-Based 54:63 arg1 Preparation 72:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 2 47 theme N-linked 227:234 arg1 moieties 243:250 The N-linked glycan moieties 223:250 The N-linked glycan moieties of these entities 223:268 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 9 48 theme sample 1489:1494 arg1 preparation 1496:1506 fast, easily automated sample preparation 1466:1506 fast, easily automated sample preparation 1466:1506 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 4 49 theme structure 669:677 arg1 alterations 679:689 disease-related glycan structure alterations 646:689 disease-related glycan structure alterations 646:689 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 5 50 theme analysis 864:871 arg1 methods 873:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 6 51 theme based 996:1000 arg1 preparation 1022:1032 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 10 52 theme glycans 1635:1641 arg1 separation 1604:1613 Rapid separation 1598:1613 Rapid separation of APTS labeled IgG glycans 1598:1641 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 7 53 theme fluorescence 1207:1218 arg1 CE-LIF 1231:1236 CE-LIF 1231:1236 CE-LIF 1231:1236 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 7 53 theme fluorescence 1207:1218 arg1 detection 1220:1228 laser induced fluorescence detection 1193:1228 laser induced fluorescence detection (CE-LIF) 1193:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 6 54 theme magnetic 982:989 arg1 preparation 1022:1032 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 10 55 theme optimized 1670:1678 arg1 protocol 1687:1694 an optimized CE-LIF protocol 1667:1694 an optimized CE-LIF protocol 1667:1694 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 2 56 theme receptor-binding 326:341 arg1 activity 343:350 receptor-binding activity 326:350 receptor-binding activity 326:350 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 9 57 theme single 1528:1533 arg1 individual 1516:1525 individual 1516:1525 individual 1516:1525 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 9 57 theme single 1528:1533 arg1 vial 1535:1538 single vial 1528:1538 single vial 1528:1538 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 4 58 theme disease-related 646:660 arg1 alterations 679:689 disease-related glycan structure alterations 646:689 disease-related glycan structure alterations 646:689 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 7 59 theme samples 1182:1188 arg1 analysis 1157:1164 rapid capillary electrophoresis analysis 1125:1164 rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF) 1125:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 9 60 theme 96-well 1545:1551 arg1 formats 1559:1565 individual (single vial) and 96-well plate formats 1516:1565 formats 1559:1565 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 5 61 gly glycoproteins 907:919 arg1 glycoproteins 907:919 glycoproteins 907:919 glycoproteins 907:919 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 1 62 theme new-generation 201:214 arg1 drugs 216:220 these promising new-generation drugs 185:220 these promising new-generation drugs 185:220 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 2 63 dep circulation 284:294 arg1 half-life 296:304 half-life 296:304 half-life 296:304 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 5 64 theme glycoproteins 907:919 arg1 profiling 894:902 N-glycan profiling 885:902 N-glycan profiling of glycoproteins 885:919 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 8 65 theme PNGase 1286:1291 arg1 F 1293:1293 the endoglycosidase PNGase F 1266:1293 the endoglycosidase PNGase F 1266:1293 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 8 66 theme charged 1347:1353 arg1 dye 1367:1369 the charged fluorophore dye 1343:1369 the charged fluorophore dye of aminopyrenetrisulfonate (APTS) 1343:1403 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 7 67 theme capillary 1131:1139 arg1 electrophoresis 1141:1155 rapid capillary electrophoresis 1125:1155 rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF) 1125:1237 A detailed protocol of this method is given in this chapter in conjunction with rapid capillary electrophoresis analysis of the prepared samples by laser induced fluorescence detection (CE-LIF). 27743372 2 68 theme entities 261:268 arg1 moieties 243:250 The N-linked glycan moieties 223:250 The N-linked glycan moieties of these entities 223:268 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 1 69 theme drugs 216:220 arg1 efficacy 173:180 efficacy 173:180 efficacy 173:180 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 1 69 theme drugs 216:220 arg1 activity 160:167 activity 160:167 activity 160:167 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 9 70 theme dye 1585:1587 arg1 removal 1589:1595 excess dye removal 1578:1595 excess dye removal 1578:1595 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 8 71 theme released 1303:1310 arg1 carbohydrates 1312:1324 the released carbohydrates 1299:1324 the released carbohydrates 1299:1324 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 6 72 theme sample 1015:1020 arg1 preparation 1022:1032 the magnetic bead based glycoprotein sample preparation 978:1032 the magnetic bead based glycoprotein sample preparation technique 978:1042 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 8 73 theme aminopyrenetrisulfonate 1374:1396 arg1 dye 1367:1369 the charged fluorophore dye 1343:1369 the charged fluorophore dye of aminopyrenetrisulfonate (APTS) 1343:1403 N-glycans are digested by the endoglycosidase PNGase F and the released carbohydrates are labeled with the charged fluorophore dye of aminopyrenetrisulfonate (APTS). 27743372 9 74 dep individual 1516:1525 arg1 both 1508:1511 both 1508:1511 both 1508:1511 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 5 75 theme applicable 813:822 arg1 methods 873:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 4 76 theme early 748:752 arg1 diagnostics 754:764 early diagnostics 748:764 early diagnostics 748:764 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 5 77 theme great 789:793 arg1 demand 795:800 a great demand 787:800 a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins 787:919 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 9 78 theme Effective 1406:1414 arg1 capture 1423:1429 Effective glycan capture 1406:1429 Effective glycan capture by magnetic microparticles 1406:1456 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 1 79 theme N-glycan 85:92 arg1 profiling 94:102 N-glycan profiling 85:102 N-glycan profiling of therapeutic glycoproteins 85:131 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 9 80 dep fast 1466:1469 arg1 automated 1479:1487 automated 1479:1487 automated 1479:1487 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 3 81 theme dollar 511:516 arg1 business 528:535 multibillion dollar worldwide business 498:535 multibillion dollar worldwide business 498:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 0 82 theme N-Glycan 16:23 arg1 Analysis 25:32 High-Throughput N-Glycan Analysis 0:32 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation 0:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 9 83 theme magnetic 1434:1441 arg1 microparticles 1443:1456 magnetic microparticles 1434:1456 magnetic microparticles 1434:1456 Effective glycan capture by magnetic microparticles enabled fast, easily automated sample preparation both in individual (single vial) and 96-well plate formats, including excess dye removal. 27743372 1 84 theme glycoproteins 119:131 arg1 profiling 94:102 N-glycan profiling 85:102 N-glycan profiling of therapeutic glycoproteins 85:131 N-glycan profiling of therapeutic glycoproteins is essential to ensure the activity and efficacy of these promising new-generation drugs. 27743372 3 85 theme more 426:429 arg1 glycoproteins 471:483 glycoproteins 471:483 glycoproteins representing multibillion dollar worldwide business 471:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 3 85 theme more 426:429 arg1 half 436:439 more than half 426:439 more than half of the biopharmaceuticals 426:465 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 5 86 theme preparation 848:858 arg1 methods 873:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 widely applicable, high-throughput sample preparation and analysis methods 806:879 Therefore, there is a great demand for widely applicable, high-throughput sample preparation and analysis methods for N-glycan profiling of glycoproteins. 27743372 6 87 theme exciting 940:947 arg1 developments 949:960 the newest exciting developments 929:960 the newest exciting developments of the field 929:973 One of the newest exciting developments of the field is the magnetic bead based glycoprotein sample preparation technique. 27743372 4 88 theme biomedical 599:608 arg1 field 610:614 the biomedical field 595:614 the biomedical field 595:614 In the biomedical field, on the other hand, revealing disease-related glycan structure alterations holds the promise of the discovery of new biomarkers for early diagnostics. 27743372 2 89 link N-linked 227:234 arg1 moieties 243:250 The N-linked glycan moieties 223:250 The N-linked glycan moieties of these entities 223:268 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 0 90 theme Sample 65:70 arg1 Preparation 72:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 Rapid Magnetic Bead-Based Sample Preparation 39:82 High-Throughput N-Glycan Analysis with Rapid Magnetic Bead-Based Sample Preparation. 27743372 10 91 theme APTS 1618:1621 arg1 glycans 1635:1641 APTS labeled IgG glycans 1618:1641 APTS labeled IgG glycans 1618:1641 Rapid separation of APTS labeled IgG glycans is also shown utilizing an optimized CE-LIF protocol. 27743372 2 92 theme glycan 236:241 arg1 moieties 243:250 The N-linked glycan moieties 223:250 The N-linked glycan moieties of these entities 223:268 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 2 93 theme thermal 383:389 arg1 properties 401:410 physicochemical and thermal stability properties 363:410 physicochemical and thermal stability properties 363:410 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 3 94 from glycoproteins 471:483 arg1 addition 416:423 addition 416:423 addition 416:423 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 2 95 theme physicochemical 363:377 arg1 properties 401:410 physicochemical and thermal stability properties 363:410 physicochemical and thermal stability properties 363:410 The N-linked glycan moieties of these entities highly affect circulation half-life, immunogenicity and receptor-binding activity as well as physicochemical and thermal stability properties. 27743372 3 96 gly glycoproteins 471:483 arg1 glycoproteins 471:483 glycoproteins 471:483 glycoproteins representing multibillion dollar worldwide business 471:535 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 27743372 3 96 gly glycoproteins 471:483 arg1 half 436:439 more than half 426:439 more than half of the biopharmaceuticals 426:465 In addition, more than half of the biopharmaceuticals are glycoproteins representing multibillion dollar worldwide business, further emphasizing the importance of their analysis. 29183914 3 0 theme specific 249:256 arg1 sequences 267:275 specific O-glycan sequences 249:275 specific O-glycan sequences 249:275 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 5 1 theme facile 579:584 arg1 resolution 602:611 facile chromatographic resolution 579:611 facile chromatographic resolution 579:611 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 6 2 theme magnitude 776:784 arg1 orders 766:771 orders 766:771 orders of magnitude more sensitive than traditional methods 766:824 This is orders of magnitude more sensitive than traditional methods. 29183914 3 3 theme O-glycan 258:265 arg1 sequences 267:275 specific O-glycan sequences 249:275 specific O-glycan sequences 249:275 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 8 4 from structures 1191:1200 arg1 microenvironments 1213:1229 natural microenvironments 1205:1229 natural microenvironments 1205:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 5 5 theme arraying 628:635 arg1 O-glycans 637:645 arraying O-glycans 628:645 arraying O-glycans as branches 628:657 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 8 6 theme wide 1116:1119 arg1 range 1121:1125 a wide range 1114:1125 a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments 1114:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 7 7 theme rotaviruses 1038:1048 arg1 proteins 1022:1029 the adhesive proteins 1009:1029 the adhesive proteins of two rotaviruses 1009:1048 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 4 8 dep approach 418:425 arg1 search 443:448 O-glycome beam search 428:448 a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery 396:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 4 9 theme beam 438:441 arg1 search 443:448 O-glycome beam search 428:448 a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery 396:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 3 10 theme methodologies 364:376 arg1 lack 346:349 a lack 344:349 a lack of efficient methodologies 344:376 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 4 11 theme O-glycan 488:495 arg1 ligand-discovery 497:512 O-glycan ligand-discovery 488:512 O-glycan ligand-discovery 488:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 3 12 theme many 280:283 arg1 systems 301:307 many bio-recognition systems 280:307 many bio-recognition systems 280:307 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 4 13 theme O-glycome 428:436 arg1 search 443:448 O-glycome beam search 428:448 a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery 396:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 8 14 theme glycan 1172:1177 arg1 structures 1191:1200 glycan recognition structures 1172:1200 glycan recognition structures in natural microenvironments 1172:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 7 15 theme undetected 942:951 arg1 components 920:929 extremely minor components 904:929 extremely minor components previously undetected 904:951 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 3 16 theme bio-recognition 285:299 arg1 systems 301:307 many bio-recognition systems 280:307 many bio-recognition systems 280:307 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 7 17 theme minor 914:918 arg1 components 920:929 extremely minor components 904:929 extremely minor components previously undetected 904:951 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 0 18 theme Beam 10:13 arg1 Arrays 22:27 O-Glycome Beam Search Arrays 0:27 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 0:62 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 29183914 3 19 theme sequences 267:275 arg1 involvement 234:244 The involvement 230:244 The involvement of specific O-glycan sequences in many bio-recognition systems 230:307 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 1 20 from mechanisms 150:159 arg1 health 164:169 health 164:169 health 164:169 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 1 20 from mechanisms 150:159 arg1 disease 175:181 disease 175:181 disease 175:181 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 1 21 theme post-translational 85:102 arg1 modification 104:115 a post-translational modification 83:115 a post-translational modification of proteins crucial to molecular mechanisms in health and disease 83:181 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 1 21 theme post-translational 85:102 arg1 O-glycosylation 64:78 O-glycosylation 64:78 O-glycosylation 64:78 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 5 22 theme complex 549:555 arg1 profile 567:573 the complex O-glycome profile 545:573 the complex O-glycome profile 545:573 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 0 23 theme O-Glycome 0:8 arg1 Arrays 22:27 O-Glycome Beam Search Arrays 0:27 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 0:62 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 29183914 7 24 theme porcine 867:873 arg1 mucin 883:887 porcine stomach mucin 867:887 porcine stomach mucin 867:887 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 8 25 theme biological 1130:1139 arg1 settings 1141:1148 biological settings 1130:1148 biological settings 1130:1148 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 4 26 theme microarray 407:416 arg1 robust 463:468 robust 463:468 robust 463:468 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 4 26 theme microarray 407:416 arg1 approach 418:425 a targeted microarray approach 396:425 a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery 396:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 5 27 theme mass 674:677 arg1 spectrometry 679:690 mass spectrometry 674:690 mass spectrometry 674:690 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 5 28 theme promising 705:713 arg1 fractions 715:723 promising fractions 705:723 promising fractions 705:723 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 7 29 theme stomach 875:881 arg1 mucin 883:887 porcine stomach mucin 867:887 porcine stomach mucin 867:887 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 4 30 theme targeted 398:405 arg1 robust 463:468 robust 463:468 robust 463:468 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 4 30 theme targeted 398:405 arg1 approach 418:425 a targeted microarray approach 396:425 a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery 396:512 We describe here a targeted microarray approach: O-glycome beam search that is both robust and efficient for O-glycan ligand-discovery. 29183914 0 31 theme Search 15:20 arg1 Arrays 22:27 O-Glycome Beam Search Arrays 0:27 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 0:62 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 29183914 1 32 theme proteins 120:127 arg1 modification 104:115 a post-translational modification 83:115 a post-translational modification of proteins crucial to molecular mechanisms in health and disease 83:181 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 1 32 theme proteins 120:127 arg1 O-glycosylation 64:78 O-glycosylation 64:78 O-glycosylation 64:78 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 0 33 theme Carbohydrate 33:44 arg1 Discovery 53:61 Carbohydrate Ligand Discovery 33:61 Carbohydrate Ligand Discovery 33:61 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 29183914 8 34 from studies 1103:1109 arg1 range 1121:1125 a wide range 1114:1125 a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments 1114:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 5 35 theme Substantial 515:525 arg1 simplification 527:540 Substantial simplification 515:540 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution 515:611 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 3 36 from involvement 234:244 arg1 systems 301:307 many bio-recognition systems 280:307 many bio-recognition systems 280:307 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 5 37 theme chromatographic 586:600 arg1 resolution 602:611 facile chromatographic resolution 579:611 facile chromatographic resolution 579:611 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 6 38 theme sensitive 791:799 arg1 orders 766:771 orders 766:771 orders of magnitude more sensitive than traditional methods 766:824 This is orders of magnitude more sensitive than traditional methods. 29183914 7 39 theme beam 843:846 arg1 approach 855:862 beam search approach 843:862 beam search approach 843:862 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 8 40 theme recognition 1179:1189 arg1 structures 1191:1200 glycan recognition structures 1172:1200 glycan recognition structures in natural microenvironments 1172:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 8 41 theme natural 1205:1211 arg1 microenvironments 1213:1229 natural microenvironments 1205:1229 natural microenvironments 1205:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 5 42 theme resolution 602:611 arg1 simplification 527:540 Substantial simplification 515:540 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution 515:611 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 1 43 theme crucial 129:135 arg1 proteins 120:127 proteins 120:127 proteins crucial to molecular mechanisms in health and disease 120:181 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 0 44 theme Ligand 46:51 arg1 Discovery 53:61 Carbohydrate Ligand Discovery 33:61 Carbohydrate Ligand Discovery 33:61 O-Glycome Beam Search Arrays for Carbohydrate Ligand Discovery. 29183914 5 45 theme on-array 730:737 arg1 immuno-sequencing 739:755 on-array immuno-sequencing 730:755 on-array immuno-sequencing 730:755 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 8 46 theme O-glycome 1081:1089 arg1 studies 1103:1109 O-glycome recognition studies 1081:1109 O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments 1081:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 6 47 theme traditional 806:816 arg1 methods 818:824 traditional methods 806:824 traditional methods 806:824 This is orders of magnitude more sensitive than traditional methods. 29183914 1 48 theme molecular 140:148 arg1 mechanisms 150:159 molecular mechanisms 140:159 molecular mechanisms in health and disease 140:181 O-glycosylation is a post-translational modification of proteins crucial to molecular mechanisms in health and disease. 29183914 5 49 theme O-glycome 557:565 arg1 profile 567:573 the complex O-glycome profile 545:573 the complex O-glycome profile 545:573 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 7 50 theme adhesive 1013:1020 arg1 proteins 1022:1029 the adhesive proteins 1009:1029 the adhesive proteins of two rotaviruses 1009:1048 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 8 51 theme recognition 1091:1101 arg1 studies 1103:1109 O-glycome recognition studies 1081:1109 O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments 1081:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 5 52 theme profile 567:573 arg1 simplification 527:540 Substantial simplification 515:540 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution 515:611 Substantial simplification of the complex O-glycome profile and facile chromatographic resolution is achieved by arraying O-glycans as branches, monitoring by mass spectrometry, focusing on promising fractions, and on-array immuno-sequencing. 29183914 7 53 theme mucin 982:986 arg1 O-glycome 964:972 the O-glycome 960:972 the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses 960:1048 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 7 53 theme mucin 982:986 arg1 ligands 997:1003 ligands 997:1003 ligands for the adhesive proteins of two rotaviruses 997:1048 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 29183914 3 54 theme efficient 354:362 arg1 methodologies 364:376 efficient methodologies 354:376 efficient methodologies 354:376 The involvement of specific O-glycan sequences in many bio-recognition systems is yet to be determined because of a lack of efficient methodologies. 29183914 8 55 theme settings 1141:1148 arg1 range 1121:1125 a wide range 1114:1125 a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments 1114:1229 The approach is applicable to O-glycome recognition studies in a wide range of biological settings to give insights into glycan recognition structures in natural microenvironments. 29183914 7 56 theme search 848:853 arg1 approach 855:862 beam search approach 843:862 beam search approach 843:862 We have applied beam search approach to porcine stomach mucin and identified extremely minor components previously undetected within the O-glycome of this mucin that are ligands for the adhesive proteins of two rotaviruses. 28737213 1 0 dep mammals 93:99 arg1 contrast 81:88 contrast 81:88 contrast 81:88 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 3 1 theme green 520:524 arg1 braunii 550:556 green microalgae Botryococcus braunii 520:556 green microalgae Botryococcus braunii 520:556 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 4 2 theme N-glycopeptides 591:605 arg1 analysis 572:579 the analysis 568:579 the analysis of intact N-glycopeptides 568:605 Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions. 28737213 0 3 from Identification 0:13 arg1 brauni 70:75 Botryococcus brauni 57:75 Botryococcus brauni 57:75 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 8 4 theme N-glycans 1109:1117 arg1 identification 1076:1089 The identification 1072:1089 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii 1072:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 5 5 theme identified 774:783 arg1 N-glycoproteins 785:799 the identified N-glycoproteins 770:799 the identified N-glycoproteins 770:799 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 0 6 from brauni 70:75 arg1 Identification 0:13 Identification 0:13 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 0:76 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 5 7 theme N-glycosylation 698:712 arg1 role 690:693 the role 686:693 the role of N-glycosylation in B. braunii 686:726 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 2 8 theme several 357:363 arg1 species 371:377 several algal species 357:377 several algal species 357:377 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 5 9 theme N-glycoproteins 785:799 arg1 annotation 756:765 functional annotation 745:765 functional annotation of the identified N-glycoproteins 745:799 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 6 10 theme nonreducing 951:961 arg1 end 963:965 the nonreducing end 947:965 the nonreducing end 947:965 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 1 11 from diversity 145:153 arg1 structures 171:180 the N-glycan structures 158:180 the N-glycan structures of complex N-glycoproteins 158:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 1 12 theme N-glycan 162:169 arg1 structures 171:180 the N-glycan structures 158:180 the N-glycan structures of complex N-glycoproteins 158:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 8 13 theme GnTI-dependent 1094:1107 arg1 N-glycans 1109:1117 GnTI-dependent N-glycans 1094:1117 GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii 1094:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 1 14 gly N-glycoproteins 193:207 arg1 N-glycoproteins 193:207 complex N-glycoproteins 185:207 complex N-glycoproteins 185:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 4 15 gly N-glycopeptides 591:605 arg2 N-glycopeptides 591:605 intact N-glycopeptides 584:605 intact N-glycopeptides 584:605 Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions. 28737213 2 16 theme key 283:285 arg1 I 271:271 β1,2-N-acetylglucosaminyltransferase I 234:271 β1,2-N-acetylglucosaminyltransferase I (GnTI) 234:278 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 2 16 theme key 283:285 arg1 enzyme 287:292 a key enzyme 281:292 a key enzyme in the formation of complex N-glycans 281:330 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 0 17 from N-glycans 44:52 arg1 brauni 70:75 Botryococcus brauni 57:75 Botryococcus brauni 57:75 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 8 18 theme N-glycan 1139:1146 arg1 methylation 1148:1158 N-glycan methylation 1139:1158 N-glycan methylation in B. braunii 1139:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 2 19 theme N-glycans 322:330 arg1 formation 301:309 the formation 297:309 the formation of complex N-glycans 297:330 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 6 20 gly N-glycopeptides 886:900 arg2 N-glycopeptides 886:900 intact N-glycopeptides 879:900 intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end 879:965 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 2 21 theme GnTI-dependent 380:393 arg1 N-glycans 395:403 GnTI-dependent N-glycans 380:403 GnTI-dependent N-glycans 380:403 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 8 22 from N-glycans 1109:1117 arg1 combination 1122:1132 combination 1122:1132 combination with N-glycan methylation in B. braunii 1122:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 6 23 theme N-glycosylated 823:836 arg1 peptides 838:845 517 unique N-glycosylated peptides 812:845 517 unique N-glycosylated peptides 812:845 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 2 24 theme complex 314:320 arg1 N-glycans 322:330 complex N-glycans 314:330 complex N-glycans 314:330 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 1 25 theme complex 185:191 arg1 N-glycoproteins 193:207 complex N-glycoproteins 185:207 complex N-glycoproteins 185:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 0 26 theme GnTI-dependent 29:42 arg1 N-glycans 44:52 methylated GnTI-dependent N-glycans 18:52 methylated GnTI-dependent N-glycans in Botryococcus brauni 18:75 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 7 27 theme GnTI-dependent 988:1001 arg1 N-glycans 1003:1011 these GnTI-dependent N-glycans 982:1011 these GnTI-dependent N-glycans 982:1011 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 6 28 theme unique 816:821 arg1 peptides 838:845 517 unique N-glycosylated peptides 812:845 517 unique N-glycosylated peptides 812:845 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 1 29 theme vascular 105:112 arg1 plants 114:119 vascular plants 105:119 vascular plants 105:119 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 1 30 theme N-glycoproteins 193:207 arg1 structures 171:180 the N-glycan structures 158:180 the N-glycan structures of complex N-glycoproteins 158:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 0 31 theme methylated 18:27 arg1 N-glycans 44:52 methylated GnTI-dependent N-glycans 18:52 methylated GnTI-dependent N-glycans in Botryococcus brauni 18:75 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 4 32 theme intact 584:589 arg1 N-glycopeptides 591:605 intact N-glycopeptides 584:605 intact N-glycopeptides 584:605 Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions. 28737213 5 33 theme functional 745:754 arg1 annotation 756:765 functional annotation 745:765 functional annotation of the identified N-glycoproteins 745:799 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 5 34 gly N-glycoproteins 785:799 arg1 N-glycoproteins 785:799 the identified N-glycoproteins 770:799 the identified N-glycoproteins 770:799 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 8 35 with combination 1122:1132 arg1 methylation 1148:1158 N-glycan methylation 1139:1158 N-glycan methylation in B. braunii 1139:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 3 36 theme microalgae 526:535 arg1 braunii 550:556 green microalgae Botryococcus braunii 520:556 green microalgae Botryococcus braunii 520:556 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 6 37 gly N-glycosylated 823:836 arg1 peptides 838:845 517 unique N-glycosylated peptides 812:845 517 unique N-glycosylated peptides 812:845 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 8 38 from identification 1076:1089 arg1 combination 1122:1132 combination 1122:1132 combination with N-glycan methylation in B. braunii 1122:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 0 39 theme N-glycans 44:52 arg1 Identification 0:13 Identification 0:13 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 0:76 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 8 40 from type 1195:1198 arg1 microalgae 1231:1240 this microalgae 1226:1240 this microalgae 1226:1240 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 8 41 theme uncommon 1186:1193 arg1 type 1195:1198 an uncommon type 1183:1198 an uncommon type of N-glycan processing in this microalgae 1183:1240 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 2 42 theme β1,2-N-acetylglucosaminyltransferase 234:269 arg1 I 271:271 β1,2-N-acetylglucosaminyltransferase I 234:271 β1,2-N-acetylglucosaminyltransferase I (GnTI) 234:278 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 2 42 theme β1,2-N-acetylglucosaminyltransferase 234:269 arg1 enzyme 287:292 a key enzyme 281:292 a key enzyme in the formation of complex N-glycans 281:330 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 2 42 theme β1,2-N-acetylglucosaminyltransferase 234:269 arg1 GnTI 274:277 GnTI 274:277 GnTI 274:277 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 3 43 theme hydrocarbon 490:500 arg1 oils 502:505 the hydrocarbon oils 486:505 the hydrocarbon oils 486:505 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 0 44 theme Botryococcus 57:68 arg1 brauni 70:75 Botryococcus brauni 57:75 Botryococcus brauni 57:75 Identification of methylated GnTI-dependent N-glycans in Botryococcus brauni. 28737213 8 45 from combination 1122:1132 arg1 identification 1076:1089 The identification 1072:1089 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii 1072:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 7 46 theme methylated 1053:1062 arg1 di 1050:1051 di 1050:1051 di 1050:1051 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 7 46 theme methylated 1053:1062 arg1 hexose 1064:1069 methylated hexose 1053:1069 (di)methylated hexose 1049:1069 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 3 47 theme oils 502:505 arg1 analysis 474:481 an N-glycoproteomic analysis 454:481 an N-glycoproteomic analysis of the hydrocarbon oils 454:505 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 2 48 from enzyme 287:292 arg1 formation 301:309 the formation 297:309 the formation of complex N-glycans 297:330 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 5 49 from role 690:693 arg1 B. braunii 717:726 B. braunii 717:726 B. braunii 717:726 Furthermore, insights into the role of N-glycosylation in B. braunii were gained from functional annotation of the identified N-glycoproteins. 28737213 4 50 theme N-glycan 636:643 arg1 compositions 645:656 N-glycan compositions 636:656 N-glycan compositions 636:656 Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions. 28737213 3 51 theme Botryococcus 537:548 arg1 braunii 550:556 green microalgae Botryococcus braunii 520:556 green microalgae Botryococcus braunii 520:556 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 8 52 theme processing 1212:1221 arg1 type 1195:1198 an uncommon type 1183:1198 an uncommon type of N-glycan processing in this microalgae 1183:1240 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 6 53 theme intact 879:884 arg1 N-glycopeptides 886:900 intact N-glycopeptides 879:900 intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end 879:965 In total, 517 unique N-glycosylated peptides have been identified, including intact N-glycopeptides that harbored N-acetylhexosamine (HexNAc) at the nonreducing end. 28737213 3 54 theme N-glycoproteomic 457:472 arg1 analysis 474:481 an N-glycoproteomic analysis 454:481 an N-glycoproteomic analysis of the hydrocarbon oils 454:505 We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. 28737213 1 55 theme high 140:143 arg1 diversity 145:153 a high diversity 138:153 a high diversity in the N-glycan structures of complex N-glycoproteins 138:207 In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. 28737213 8 56 theme N-glycan 1203:1210 arg1 processing 1212:1221 N-glycan processing 1203:1221 N-glycan processing 1203:1221 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 8 57 from methylation 1148:1158 arg1 B. braunii 1163:1172 B. braunii 1163:1172 B. braunii 1163:1172 The identification of GnTI-dependent N-glycans in combination with N-glycan methylation in B. braunii revealed an uncommon type of N-glycan processing in this microalgae. 28737213 7 58 mod modified 1035:1042 arg3 di 1050:1051 di 1050:1051 di 1050:1051 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 7 58 mod modified 1035:1042 arg1 N-glycans 1003:1011 these GnTI-dependent N-glycans 982:1011 these GnTI-dependent N-glycans 982:1011 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 7 58 mod modified 1035:1042 arg3 hexose 1064:1069 methylated hexose 1053:1069 (di)methylated hexose 1049:1069 Surprisingly, these GnTI-dependent N-glycans were also found to be modified with (di)methylated hexose. 28737213 2 59 theme algal 365:369 arg1 species 371:377 several algal species 357:377 several algal species 357:377 Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. 28737213 4 60 theme compositions 645:656 arg1 determination 619:631 the determination 615:631 the determination of N-glycan compositions 615:656 Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions. 29217182 3 0 attach released 422:429 arg2 IgY 365:367 IgY 365:367 IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan 365:416 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 3 0 attach released 422:429 arg1 IgY 436:438 IgY 436:438 IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment 436:513 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 0 1 theme egg 84:86 arg1 yolk 88:91 hen egg yolk 80:91 hen egg yolk 80:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 4 2 theme N-Glycosylation 658:672 arg1 ASN308 681:686 ASN308 681:686 ASN308 681:686 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 2 theme N-Glycosylation 658:672 arg1 sites 674:678 Two potential N-Glycosylation sites 644:678 Two potential N-Glycosylation sites (ASN308 and ASN409) 644:698 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 2 theme N-Glycosylation 658:672 arg1 ASN409 692:697 ASN409 692:697 ASN409 692:697 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 3 3 theme purified 312:319 arg1 protein 321:327 purified protein 312:327 purified protein from hen egg yolk 312:345 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 4 4 theme Sugar 739:743 arg1 chains 745:750 Sugar chains 739:750 Sugar chains 739:750 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 5 theme N-glycan 844:851 arg1 structures 853:862 17 N-glycan structures 841:862 17 N-glycan structures 841:862 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 6 theme liquid 786:791 arg1 chromatography 793:806 normal phase liquid chromatography 773:806 normal phase liquid chromatography 773:806 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 7 dep sites 674:678 arg1 ASN308 681:686 ASN308 681:686 ASN308 681:686 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 7 dep sites 674:678 arg1 sites 674:678 Two potential N-Glycosylation sites 644:678 Two potential N-Glycosylation sites (ASN308 and ASN409) 644:698 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 7 dep sites 674:678 arg1 ASN409 692:697 ASN409 692:697 ASN409 692:697 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 3 8 theme molecular 524:532 arg1 weight 534:539 the molecular weight 520:539 the molecular weight of IgY 520:546 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 5 9 theme chain 903:907 arg1 pattern 909:915 The sugar chain pattern 893:915 The sugar chain pattern 893:915 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 5 10 theme complex 984:990 arg1 oligosaccharide 992:1006 complex oligosaccharide 984:1006 complex oligosaccharide 984:1006 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 6 11 theme important 1043:1051 arg1 information 1053:1063 other important information 1037:1063 other important information regarding IgY glycosylation 1037:1091 These results could lead to other important information regarding IgY glycosylation. 29217182 2 12 theme N-glycan 226:233 arg1 structures 235:244 its N-glycan structures 222:244 its N-glycan structures 222:244 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 6 13 theme IgY 1075:1077 arg1 glycosylation 1079:1091 IgY glycosylation 1075:1091 IgY glycosylation 1075:1091 These results could lead to other important information regarding IgY glycosylation. 29217182 4 14 dep separated 757:765 arg1 MS. 735:737 MS. 735:737 MS. 735:737 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 0 15 theme immunoglobulin 58:71 arg1 Y 73:73 immunoglobulin Y 58:73 immunoglobulin Y from hen egg yolk 58:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 5 16 theme high-mannose 927:938 arg1 oligosaccharide 940:954 high-mannose oligosaccharide 927:954 high-mannose oligosaccharide 927:954 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 3 17 theme egg 338:340 arg1 yolk 342:345 hen egg yolk 334:345 hen egg yolk 334:345 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 0 18 from N-glycosylation 39:53 arg1 yolk 88:91 hen egg yolk 80:91 hen egg yolk 80:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 2 19 theme site-specific 266:278 arg1 N-glycosylation 280:294 site-specific N-glycosylation 266:294 site-specific N-glycosylation 266:294 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 1 20 theme hen 162:164 arg1 yolk 170:173 hen egg yolk 162:173 hen egg yolk 162:173 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 0 21 theme Mass 0:3 arg1 spectrometry 5:16 Mass spectrometry 0:16 Mass spectrometry 0:16 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 4 22 theme fluorescence 814:825 arg1 labeling 827:834 fluorescence labeling 814:834 fluorescence labeling 814:834 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 1 23 theme Immunoglobulin 94:107 arg1 antibody 138:145 a new therapeutic antibody 120:145 a new therapeutic antibody that exists in hen egg yolk 120:173 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 1 23 theme Immunoglobulin 94:107 arg1 IgY 112:114 IgY 112:114 IgY 112:114 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 1 23 theme Immunoglobulin 94:107 arg1 Y 109:109 Immunoglobulin Y 94:109 Immunoglobulin Y (IgY) 94:115 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 1 24 theme egg 166:168 arg1 yolk 170:173 hen egg yolk 162:173 hen egg yolk 162:173 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 6 25 theme other 1037:1041 arg1 information 1053:1063 other important information 1037:1063 other important information regarding IgY glycosylation 1037:1091 These results could lead to other important information regarding IgY glycosylation. 29217182 5 26 contain contained 917:925 arg2 oligosaccharide 964:978 hybrid oligosaccharide 957:978 hybrid oligosaccharide 957:978 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 5 26 contain contained 917:925 arg2 oligosaccharide 992:1006 complex oligosaccharide 984:1006 complex oligosaccharide 984:1006 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 5 26 contain contained 917:925 arg1 pattern 909:915 The sugar chain pattern 893:915 The sugar chain pattern 893:915 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 5 26 contain contained 917:925 arg2 oligosaccharide 940:954 high-mannose oligosaccharide 927:954 high-mannose oligosaccharide 927:954 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 2 27 dep glycoprotein 184:195 arg1 known 210:214 known 210:214 is known about its N-glycan structures, site occupancy and site-specific N-glycosylation 207:294 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 3 28 theme hen 334:336 arg1 yolk 342:345 hen egg yolk 334:345 hen egg yolk 334:345 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 3 29 theme asparagine-amidase 486:503 arg1 treatment 505:513 peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment 446:513 peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment 446:513 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 0 30 from yolk 88:91 arg1 N-glycosylation 39:53 N-glycosylation 39:53 N-glycosylation of immunoglobulin Y from hen egg yolk 39:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 0 30 from yolk 88:91 arg1 Y 73:73 immunoglobulin Y 58:73 immunoglobulin Y from hen egg yolk 58:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 2 31 theme site 247:250 arg1 occupancy 252:260 site occupancy 247:260 site occupancy 247:260 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 4 32 theme potential 648:656 arg1 ASN308 681:686 ASN308 681:686 ASN308 681:686 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 32 theme potential 648:656 arg1 sites 674:678 Two potential N-Glycosylation sites 644:678 Two potential N-Glycosylation sites (ASN308 and ASN409) 644:698 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 32 theme potential 648:656 arg1 ASN409 692:697 ASN409 692:697 ASN409 692:697 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 3 33 theme IgY 616:618 arg1 weight 606:611 the molecular weight 592:611 the molecular weight of IgY and deglycosylated IgY 592:641 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 4 34 theme normal 773:778 arg1 chromatography 793:806 normal phase liquid chromatography 773:806 normal phase liquid chromatography 773:806 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 3 35 theme MALDI-TOF/TOF 391:403 arg1 N-glycan 409:416 MALDI-TOF/TOF MS. N-glycan 391:416 MALDI-TOF/TOF MS. N-glycan 391:416 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 3 36 theme molecular 596:604 arg1 weight 606:611 the molecular weight 592:611 the molecular weight of IgY and deglycosylated IgY 592:641 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 3 37 theme MS. 405:407 arg1 N-glycan 409:416 MALDI-TOF/TOF MS. N-glycan 391:416 MALDI-TOF/TOF MS. N-glycan 391:416 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 0 38 theme Y 73:73 arg1 N-glycosylation 39:53 N-glycosylation 39:53 N-glycosylation of immunoglobulin Y from hen egg yolk 39:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 5 39 theme hybrid 957:962 arg1 oligosaccharide 964:978 hybrid oligosaccharide 957:978 hybrid oligosaccharide 957:978 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 3 40 theme IgY 544:546 arg1 weight 534:539 the molecular weight 520:539 the molecular weight of IgY 520:546 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 2 41 gly glycoprotein 184:195 arg1 It 176:177 It 176:177 It 176:177 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 2 41 gly glycoprotein 184:195 arg1 glycoprotein 184:195 a glycoprotein 182:195 a glycoprotein 182:195 It is a glycoprotein, not much is known about its N-glycan structures, site occupancy and site-specific N-glycosylation. 29217182 4 42 located detected 705:712 arg2 ASN409 692:697 ASN409 692:697 ASN409 692:697 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 42 located detected 705:712 arg2 ASN308 681:686 ASN308 681:686 ASN308 681:686 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 42 located detected 705:712 arg2 sites 674:678 Two potential N-Glycosylation sites 644:678 Two potential N-Glycosylation sites (ASN308 and ASN409) 644:698 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 4 42 located detected 705:712 arg1 IgY 717:719 IgY 717:719 IgY 717:719 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 3 43 theme IgY 639:641 arg1 weight 606:611 the molecular weight 592:611 the molecular weight of IgY and deglycosylated IgY 592:641 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 4 44 theme phase 780:784 arg1 chromatography 793:806 normal phase liquid chromatography 773:806 normal phase liquid chromatography 773:806 Two potential N-Glycosylation sites (ASN308 and ASN409) were detected on IgY by nanoLC-ESI MS. Sugar chains were separated using normal phase liquid chromatography after fluorescence labeling, and 17 N-glycan structures were confirmed using ESI-MS. 29217182 0 45 gly N-glycosylation 39:53 arg1 Y 73:73 immunoglobulin Y 58:73 immunoglobulin Y from hen egg yolk 58:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 3 46 from yolk 342:345 arg1 protein 321:327 purified protein 312:327 purified protein from hen egg yolk 312:345 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 5 47 theme sugar 897:901 arg1 pattern 909:915 The sugar chain pattern 893:915 The sugar chain pattern 893:915 The sugar chain pattern contained high-mannose oligosaccharide, hybrid oligosaccharide and complex oligosaccharide. 29217182 0 48 theme hen 80:82 arg1 yolk 88:91 hen egg yolk 80:91 hen egg yolk 80:91 Mass spectrometry characterization for N-glycosylation of immunoglobulin Y from hen egg yolk. 29217182 3 49 theme deglycosylated 624:637 arg1 IgY 639:641 deglycosylated IgY 624:641 deglycosylated IgY 624:641 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 1 50 theme new 122:124 arg1 Y 109:109 Immunoglobulin Y 94:109 Immunoglobulin Y (IgY) 94:115 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 1 50 theme new 122:124 arg1 antibody 138:145 a new therapeutic antibody 120:145 a new therapeutic antibody that exists in hen egg yolk 120:173 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 3 51 gly deglycosylated 624:637 arg1 IgY 639:641 deglycosylated IgY 624:641 deglycosylated IgY 624:641 In this study, purified protein from hen egg yolk was identified as IgY based on SDS-PAGE and MALDI-TOF/TOF MS. N-glycan was released from IgY using peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase treatment, and the molecular weight of IgY was calculated using the difference between the molecular weight of IgY and deglycosylated IgY. 29217182 1 52 theme therapeutic 126:136 arg1 Y 109:109 Immunoglobulin Y 94:109 Immunoglobulin Y (IgY) 94:115 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 29217182 1 52 theme therapeutic 126:136 arg1 antibody 138:145 a new therapeutic antibody 120:145 a new therapeutic antibody that exists in hen egg yolk 120:173 Immunoglobulin Y (IgY) is a new therapeutic antibody that exists in hen egg yolk. 27459092 0 0 theme avian 87:91 arg1 system 100:105 the avian immune system 83:105 the avian immune system 83:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 4 1 theme high 1071:1074 arg1 mannose 1076:1082 high mannose 1071:1082 high mannose 1071:1082 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 0 2 theme antibodies 189:198 arg1 generation 147:156 the generation 143:156 the generation of high-quality, high-affinity antibodies to a wide range of antigens 143:226 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 7 3 theme various 1451:1457 arg1 diseases 1470:1477 various infectious diseases 1451:1477 various infectious diseases 1451:1477 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 5 4 theme sialic 1163:1168 arg1 structures 1175:1184 rare sialic acid structures 1158:1184 rare sialic acid structures 1158:1184 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 5 5 theme polysialic 1194:1203 arg1 acids 1205:1209 likely polysialic acids 1187:1209 likely polysialic acids 1187:1209 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 3 6 theme chickens 752:759 arg1 serum 735:739 the serum 731:739 the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 731:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 2 7 attach present 507:513 arg2 moieties 498:505 the carbohydrate moieties 481:505 the carbohydrate moieties present on immunoglobulins 481:532 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 2 7 attach present 507:513 arg1 immunoglobulins 518:532 immunoglobulins 518:532 immunoglobulins 518:532 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 5 8 theme rare 1158:1161 arg1 structures 1175:1184 rare sialic acid structures 1158:1184 rare sialic acid structures 1158:1184 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 0 9 theme system 100:105 arg1 exploitation 67:78 Avian Immunoglobulin Y. Recent exploitation 36:78 Avian Immunoglobulin Y. Recent exploitation of the avian immune system 36:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 4 10 theme bi- 981:983 arg1 glycans 1084:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 3 11 from serum 735:739 arg1 antibodies 715:724 IgY polyclonal antibodies 700:724 IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 700:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 3 11 from serum 735:739 arg1 profile 689:695 the detailed N-glycan profile 667:695 the detailed N-glycan profile 667:695 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 3 12 theme released 902:909 arg1 glycans 911:917 released glycans 902:917 released glycans 902:917 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 13 theme immune 93:98 arg1 system 100:105 the avian immune system 83:105 the avian immune system 83:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 3 14 theme N-glycan 804:811 arg1 approach 822:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 4 15 theme complex 973:979 arg1 bi- 981:983 complex bi- 973:983 complex bi- 973:983 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 4 16 theme mannose 1076:1082 arg1 glycans 1084:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 3 17 theme glycans 911:917 arg1 separation 888:897 ultra-performance liquid chromatography (UPLC) separation 841:897 ultra-performance liquid chromatography (UPLC) separation of released glycans 841:917 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 2 18 theme glycan 558:563 arg1 structures 565:574 certain glycan structures 550:574 certain glycan structures 550:574 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 3 19 theme analysis 813:820 arg1 approach 822:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 7 20 theme infectious 1459:1468 arg1 diseases 1470:1477 various infectious diseases 1451:1477 various infectious diseases 1451:1477 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 7 21 theme immunoglobulin-based 1399:1418 arg1 therapies 1420:1428 immunoglobulin-based therapies 1399:1428 immunoglobulin-based therapies for the treatment of various infectious diseases 1399:1477 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 6 22 theme non-human 1298:1306 arg1 glycans 1323:1329 no known non-human or immunogenic glycans 1289:1329 no known non-human or immunogenic glycans 1289:1329 It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. 27459092 2 23 theme certain 550:556 arg1 structures 565:574 certain glycan structures 550:574 certain glycan structures 550:574 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 3 24 theme quantitative 775:786 arg1 approach 822:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 25 theme wide 205:208 arg1 range 210:214 a wide range 203:214 a wide range of antigens 203:226 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 3 26 from profile 689:695 arg1 serum 735:739 the serum 731:739 the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 731:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 3 27 theme ultra-performance 841:857 arg1 UPLC 882:885 UPLC 882:885 UPLC 882:885 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 3 27 theme ultra-performance 841:857 arg1 chromatography 866:879 ultra-performance liquid chromatography 841:879 ultra-performance liquid chromatography (UPLC) separation of released glycans 841:917 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 28 theme N-Glycan 14:21 arg1 Profiling 23:31 Comprehensive N-Glycan Profiling 0:31 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system 0:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 3 29 theme high-throughput 788:802 arg1 approach 822:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 a fully quantitative high-throughput N-glycan analysis approach 767:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 30 theme antigens 219:226 arg1 range 210:214 a wide range 203:214 a wide range of antigens 203:226 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 0 31 theme therapeutic 244:254 arg1 applications 277:288 therapeutic and biotechnological applications 244:288 applications 277:288 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 5 32 theme acid 1170:1173 arg1 structures 1175:1184 rare sialic acid structures 1158:1184 rare sialic acid structures 1158:1184 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 0 33 theme Comprehensive 0:12 arg1 Profiling 23:31 Comprehensive N-Glycan Profiling 0:31 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system 0:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 1 34 dep specific 369:376 arg1 specific 369:376 specific 369:376 specific 369:376 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 34 dep specific 369:376 arg1 profile 309:315 The glycosylation profile 291:315 The glycosylation profile of potential immunoglobulin therapeutics 291:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 34 dep specific 369:376 arg1 species 361:367 species 361:367 species 361:367 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 4 35 theme serum 952:956 arg1 IgY 958:960 serum IgY 952:960 serum IgY 952:960 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 0 36 theme applications 277:288 arg1 number 234:239 a number 232:239 a number of therapeutic and biotechnological applications 232:288 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 7 37 theme promising 1375:1383 arg1 candidate 1385:1393 a potentially promising candidate 1361:1393 a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases 1361:1477 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 7 37 theme promising 1375:1383 arg1 IgY 1354:1356 IgY 1354:1356 IgY 1354:1356 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 6 38 theme complex 1263:1269 arg1 glycans 1271:1277 complex glycans 1263:1277 complex glycans 1263:1277 It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. 27459092 2 39 theme physicochemical 603:617 arg1 properties 634:643 their physicochemical and biological properties 597:643 their physicochemical and biological properties 597:643 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 0 40 theme biotechnological 260:275 arg1 applications 277:288 therapeutic and biotechnological applications 244:288 applications 277:288 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 7 41 theme diseases 1470:1477 arg1 treatment 1438:1446 the treatment 1434:1446 the treatment of various infectious diseases 1434:1477 Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. 27459092 6 42 theme known 1292:1296 arg1 glycans 1323:1329 no known non-human or immunogenic glycans 1289:1329 no known non-human or immunogenic glycans 1289:1329 It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. 27459092 1 43 theme glycosylation 295:307 arg1 specific 369:376 specific 369:376 specific 369:376 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 43 theme glycosylation 295:307 arg1 profile 309:315 The glycosylation profile 291:315 The glycosylation profile of potential immunoglobulin therapeutics 291:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 43 theme glycosylation 295:307 arg1 species 361:367 species 361:367 species 361:367 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 4 44 theme glycans 1011:1017 arg1 glycans 1084:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 3 45 theme antibodies 715:724 arg1 profile 689:695 the detailed N-glycan profile 667:695 the detailed N-glycan profile 667:695 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 5 46 theme structures 1175:1184 arg1 presence 1146:1153 the presence 1142:1153 the presence of rare sialic acid structures 1142:1184 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 2 47 theme moieties 498:505 arg1 knowledge 468:476 knowledge 468:476 knowledge of the carbohydrate moieties present on immunoglobulins 468:532 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 0 48 theme Immunoglobulin 42:55 arg1 exploitation 67:78 Avian Immunoglobulin Y. Recent exploitation 36:78 Avian Immunoglobulin Y. Recent exploitation of the avian immune system 36:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 5 49 theme High 1093:1096 arg1 content 1110:1116 High sialic acid content 1093:1116 High sialic acid content 1093:1116 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 1 50 theme cell-line/culture 411:427 arg1 conditions 429:438 the cell-line/culture conditions 407:438 the cell-line/culture conditions used for production 407:458 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 5 51 theme sialic 1098:1103 arg1 content 1110:1116 High sialic acid content 1093:1116 High sialic acid content 1093:1116 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 0 52 theme Avian 36:40 arg1 exploitation 67:78 Avian Immunoglobulin Y. Recent exploitation 36:78 Avian Immunoglobulin Y. Recent exploitation of the avian immune system 36:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 3 53 theme chromatography 866:879 arg1 separation 888:897 ultra-performance liquid chromatography (UPLC) separation 841:897 ultra-performance liquid chromatography (UPLC) separation of released glycans 841:917 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 2 54 theme present 507:513 arg1 moieties 498:505 the carbohydrate moieties 481:505 the carbohydrate moieties present on immunoglobulins 481:532 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 3 55 theme IgY 700:702 arg1 antibodies 715:724 IgY polyclonal antibodies 700:724 IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 700:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 5 56 theme acid 1105:1108 arg1 content 1110:1116 High sialic acid content 1093:1116 High sialic acid content 1093:1116 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 3 57 theme liquid 859:864 arg1 UPLC 882:885 UPLC 882:885 UPLC 882:885 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 3 57 theme liquid 859:864 arg1 chromatography 866:879 ultra-performance liquid chromatography 841:879 ultra-performance liquid chromatography (UPLC) separation of released glycans 841:917 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 58 theme Recent 60:65 arg1 exploitation 67:78 Avian Immunoglobulin Y. Recent exploitation 36:78 Avian Immunoglobulin Y. Recent exploitation of the avian immune system 36:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 1 59 theme potential 320:328 arg1 therapeutics 345:356 potential immunoglobulin therapeutics 320:356 potential immunoglobulin therapeutics 320:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 4 60 theme tri- 986:989 arg1 glycans 1011:1017 tri- and tetra-antennary glycans 986:1017 tri- and tetra-antennary glycans with or without core fucose and bisects 986:1057 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 3 61 theme polyclonal 704:713 arg1 antibodies 715:724 IgY polyclonal antibodies 700:724 IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 700:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 4 62 theme Structural 920:929 arg1 assignments 931:941 Structural assignments 920:941 Structural assignments 920:941 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 4 63 theme tetra-antennary 995:1009 arg1 glycans 1011:1017 tri- and tetra-antennary glycans 986:1017 tri- and tetra-antennary glycans with or without core fucose and bisects 986:1057 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 0 64 theme Y. 57:58 arg1 exploitation 67:78 Avian Immunoglobulin Y. Recent exploitation 36:78 Avian Immunoglobulin Y. Recent exploitation of the avian immune system 36:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 1 65 theme immunoglobulin 330:343 arg1 therapeutics 345:356 potential immunoglobulin therapeutics 320:356 potential immunoglobulin therapeutics 320:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 2 66 theme carbohydrate 485:496 arg1 moieties 498:505 the carbohydrate moieties 481:505 the carbohydrate moieties present on immunoglobulins 481:532 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 6 67 theme immunogenic 1311:1321 arg1 glycans 1323:1329 no known non-human or immunogenic glycans 1289:1329 no known non-human or immunogenic glycans 1289:1329 It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. 27459092 0 68 dep high-quality 161:172 arg1 high-affinity 175:187 high-affinity 175:187 high-affinity 175:187 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 1 69 theme therapeutics 345:356 arg1 specific 369:376 specific 369:376 specific 369:376 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 69 theme therapeutics 345:356 arg1 profile 309:315 The glycosylation profile 291:315 The glycosylation profile of potential immunoglobulin therapeutics 291:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 1 69 theme therapeutics 345:356 arg1 species 361:367 species 361:367 species 361:367 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 2 70 theme biological 623:632 arg1 properties 634:643 their physicochemical and biological properties 597:643 their physicochemical and biological properties 597:643 Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. 27459092 3 71 theme leghorn 744:750 arg1 chickens 752:759 leghorn chickens 744:759 leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach 744:829 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 1 72 gly glycosylation 295:307 arg1 therapeutics 345:356 potential immunoglobulin therapeutics 320:356 potential immunoglobulin therapeutics 320:356 The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. 27459092 0 73 theme high-quality 161:172 arg1 antibodies 189:198 high-quality, high-affinity antibodies 161:198 high-quality, high-affinity antibodies to a wide range of antigens 161:226 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 4 74 theme hybrid 1060:1065 arg1 glycans 1084:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans 973:1090 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27459092 3 75 theme detailed 671:678 arg1 profile 689:695 the detailed N-glycan profile 667:695 the detailed N-glycan profile 667:695 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 0 76 theme exploitation 67:78 arg1 Profiling 23:31 Comprehensive N-Glycan Profiling 0:31 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system 0:105 Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y. Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. 27459092 5 77 theme likely 1187:1192 arg1 acids 1205:1209 likely polysialic acids 1187:1209 likely polysialic acids 1187:1209 High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. 27459092 3 78 theme N-glycan 680:687 arg1 profile 689:695 the detailed N-glycan profile 667:695 the detailed N-glycan profile 667:695 This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. 27459092 4 79 theme core 1035:1038 arg1 fucose 1040:1045 core fucose 1035:1045 core fucose 1035:1045 Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. 27058295 2 0 theme convergent 468:477 arg1 synthesis 479:487 the convergent synthesis 464:487 the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 464:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 1 1 link N-linked 225:232 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 2 2 theme acid 513:516 arg1 synthesis 479:487 the convergent synthesis 464:487 the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 464:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 3 used used 361:364 arg2 N-glycan 288:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 3 used used 361:364 arg2 substrate 377:385 a donor substrate 369:385 a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 369:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 4 theme amino 507:511 arg1 acid 513:516 a glycosylated amino acid 492:516 a glycosylated amino acid bearing a hybrid N-glycan structure 492:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 5 theme glycosylated 494:505 arg1 acid 513:516 a glycosylated amino acid 492:516 a glycosylated amino acid bearing a hybrid N-glycan structure 492:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 6 theme Endo 407:410 arg1 M 412:412 wild type Endo M 397:412 wild type Endo M 397:412 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 7 theme Endo 441:444 arg1 mutant 448:453 an N175Q glycosynthase Endo M mutant 418:453 an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 418:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 8 gly glycosylated 494:505 arg1 acid 513:516 a glycosylated amino acid 492:516 a glycosylated amino acid bearing a hybrid N-glycan structure 492:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 9 theme glycosynthase 427:439 arg1 mutant 448:453 an N175Q glycosynthase Endo M mutant 418:453 an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 418:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 1 10 theme wide 190:193 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 1 10 theme wide 190:193 arg1 variety 195:201 a wide variety 188:201 a wide variety of defined homogenous N-linked glycoconjugates 188:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 2 11 theme N175Q 421:425 arg1 mutant 448:453 an N175Q glycosynthase Endo M mutant 418:453 an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 418:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 12 theme donor 371:375 arg1 N-glycan 288:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 12 theme donor 371:375 arg1 substrate 377:385 a donor substrate 369:385 a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 369:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 0 13 theme hybrid 15:20 arg1 oxazoline 52:60 a hybrid type N-glycan heptasaccharide oxazoline 13:60 a hybrid type N-glycan heptasaccharide oxazoline for Endo M 13:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 2 14 theme N-glycan 535:542 arg1 structure 544:552 a hybrid N-glycan structure 526:552 a hybrid N-glycan structure 526:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 15 theme hybrid 528:533 arg1 structure 544:552 a hybrid N-glycan structure 526:552 a hybrid N-glycan structure 526:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 16 theme total 313:317 arg1 synthesis 319:327 total synthesis 313:327 total synthesis 313:327 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 1 17 theme defined 206:212 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 0 18 theme N-glycan 27:34 arg1 oxazoline 52:60 a hybrid type N-glycan heptasaccharide oxazoline 13:60 a hybrid type N-glycan heptasaccharide oxazoline for Endo M 13:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 1 19 theme homogenous 214:223 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 2 20 theme type 402:405 arg1 M 412:412 wild type Endo M 397:412 wild type Endo M 397:412 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 0 21 theme type 22:25 arg1 oxazoline 52:60 a hybrid type N-glycan heptasaccharide oxazoline 13:60 a hybrid type N-glycan heptasaccharide oxazoline for Endo M 13:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 1 22 theme N-linked 225:232 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 0 23 theme oxazoline 52:60 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M 0:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 0 24 theme heptasaccharide 36:50 arg1 oxazoline 52:60 a hybrid type N-glycan heptasaccharide oxazoline 13:60 a hybrid type N-glycan heptasaccharide oxazoline for Endo M 13:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 2 25 theme hybrid-type 276:286 arg1 N-glycan 288:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan 274:295 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 25 theme hybrid-type 276:286 arg1 substrate 377:385 a donor substrate 369:385 a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 369:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 1 26 theme glycoconjugates 234:248 arg1 glycoconjugates 234:248 defined homogenous N-linked glycoconjugates 206:248 defined homogenous N-linked glycoconjugates 206:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 1 26 theme glycoconjugates 234:248 arg1 variety 195:201 a wide variety 188:201 a wide variety of defined homogenous N-linked glycoconjugates 188:248 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 1 27 theme versatile 144:152 arg1 Endo-β-N-acetylglucosaminidases 98:128 Endo-β-N-acetylglucosaminidases 98:128 Endo-β-N-acetylglucosaminidases (ENGases) 98:138 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 1 27 theme versatile 144:152 arg1 biocatalysts 154:165 versatile biocatalysts 144:165 versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner 144:271 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 27058295 0 28 theme Endo 66:69 arg1 M 71:71 Endo M 66:71 Endo M 66:71 Synthesis of a hybrid type N-glycan heptasaccharide oxazoline for Endo M catalysed glycosylation. 27058295 2 29 theme wild 397:400 arg1 M 412:412 wild type Endo M 397:412 wild type Endo M 397:412 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 30 theme M 446:446 arg1 mutant 448:453 an N175Q glycosynthase Endo M mutant 418:453 an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 418:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 31 with substrate 377:385 arg1 mutant 448:453 an N175Q glycosynthase Endo M mutant 418:453 an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure 418:552 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 2 31 with substrate 377:385 arg1 M 412:412 wild type Endo M 397:412 wild type Endo M 397:412 A hybrid-type N-glycan was accessed by total synthesis, converted to an oxazoline, and used as a donor substrate with both wild type Endo M and an N175Q glycosynthase Endo M mutant allowing the convergent synthesis of a glycosylated amino acid bearing a hybrid N-glycan structure. 27058295 1 32 theme convergent 255:264 arg1 manner 266:271 a convergent manner 253:271 a convergent manner 253:271 Endo-β-N-acetylglucosaminidases (ENGases) are versatile biocatalysts that allow access to a wide variety of defined homogenous N-linked glycoconjugates in a convergent manner. 24161615 2 0 link O-linked 351:358 arg1 glycans 360:366 O-linked glycans 351:366 O-linked glycans 351:366 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 2 1 theme ammonia-based 383:395 arg1 strategies 446:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 1 2 theme glycans 259:265 arg1 analysis 238:245 quantitative and nonselective analysis 208:245 quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins 208:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 3 3 from discrepancy 472:482 arg1 activity 503:510 deglycosylation activity 487:510 deglycosylation activity 487:510 A significant discrepancy in deglycosylation activity was observed between the ammonia-based and hydrazinolysis procedures. 24161615 9 4 theme Identity 1099:1106 arg1 confirmation 1108:1119 Identity confirmation 1099:1119 Identity confirmation 1099:1119 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 0 5 theme recombinant 99:109 arg1 proteins 111:118 recombinant proteins 99:118 recombinant proteins 99:118 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 1 6 link O-linked 250:257 arg1 glycans 259:265 O-linked glycans 250:265 O-linked glycans released from therapeutic glycoproteins 250:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 9 7 theme chromatography 1189:1202 arg1 fractions 1212:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 5 8 theme species 847:853 arg1 release 818:824 the release 814:824 the release of particular glycan species 814:853 In addition, the ammonia-based reagent demonstrated bias in the release of particular glycan species. 24161615 4 9 dep 30 664:665 arg1 to 661:662 to 661:662 to 661:662 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 5 10 theme ammonia-based 771:783 arg1 reagent 785:791 the ammonia-based reagent 767:791 the ammonia-based reagent 767:791 In addition, the ammonia-based reagent demonstrated bias in the release of particular glycan species. 24161615 6 11 theme O-glycans 941:949 arg1 characterization 921:936 characterization 921:936 characterization of O-glycans 921:949 A robust quantitative hydrazinolysis procedure was developed for characterization of O-glycans. 24161615 8 12 theme quantitative 1051:1062 arg1 release 1077:1083 quantitative nonselective release 1051:1083 quantitative nonselective release of O-glycans 1051:1096 It was shown that this procedure is superior for quantitative nonselective release of O-glycans. 24161615 9 13 theme linear 1250:1255 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 13 theme linear 1250:1255 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 1 14 theme therapeutic 281:291 arg1 glycoproteins 293:305 therapeutic glycoproteins 281:305 therapeutic glycoproteins 281:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 3 15 theme deglycosylation 487:501 arg1 activity 503:510 deglycosylation activity 487:510 deglycosylation activity 487:510 A significant discrepancy in deglycosylation activity was observed between the ammonia-based and hydrazinolysis procedures. 24161615 0 16 theme proteins 111:118 arg1 characterization 79:94 characterization 79:94 characterization of recombinant proteins 79:118 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 6 17 theme hydrazinolysis 878:891 arg1 procedure 893:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure was developed for characterization of O-glycans. 24161615 2 18 theme O-linked 351:358 arg1 glycans 360:366 O-linked glycans 351:366 O-linked glycans 351:366 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 9 19 theme trap 1261:1264 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 19 theme trap 1261:1264 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 20 theme structure 1125:1133 arg1 elucidation 1135:1145 structure elucidation 1125:1145 structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions 1125:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 21 from elucidation 1135:1145 arg1 fractions 1212:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 22 dep spectrometry 1289:1300 arg1 transform 1274:1282 transform 1274:1282 transform 1274:1282 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 0 23 link O-linked 46:53 arg1 analysis 62:69 quantitative O-linked glycan analysis 33:69 quantitative O-linked glycan analysis 33:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 3 24 theme significant 460:470 arg1 discrepancy 472:482 A significant discrepancy 458:482 A significant discrepancy in deglycosylation activity 458:510 A significant discrepancy in deglycosylation activity was observed between the ammonia-based and hydrazinolysis procedures. 24161615 9 25 theme Fourier 1266:1272 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 25 theme Fourier 1266:1272 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 2 26 theme deglycosylation 430:444 arg1 strategies 446:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 9 27 from fractions 1212:1220 arg1 O-glycans 1150:1158 O-glycans 1150:1158 O-glycans from hydrophilic interaction chromatography (HILIC) fractions 1150:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 27 from fractions 1212:1220 arg1 confirmation 1108:1119 Identity confirmation 1099:1119 Identity confirmation 1099:1119 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 27 from fractions 1212:1220 arg1 elucidation 1135:1145 structure elucidation 1125:1145 structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions 1125:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 5 28 theme glycan 840:845 arg1 species 847:853 particular glycan species 829:853 particular glycan species 829:853 In addition, the ammonia-based reagent demonstrated bias in the release of particular glycan species. 24161615 2 29 theme hydrazinolysis 415:428 arg1 strategies 446:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 0 30 theme approaches 18:27 arg1 Comparison 0:9 Comparison 0:9 Comparison of two approaches for quantitative O-linked glycan analysis 0:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 2 31 theme quantitative 327:338 arg1 release 340:346 quantitative release 327:346 quantitative release of O-linked glycans 327:366 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 9 32 theme mass 1284:1287 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 32 theme mass 1284:1287 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 3 33 theme ammonia-based 537:549 arg1 procedures 570:579 the ammonia-based and hydrazinolysis procedures 533:579 procedures 570:579 A significant discrepancy in deglycosylation activity was observed between the ammonia-based and hydrazinolysis procedures. 24161615 9 34 theme O-glycans 1150:1158 arg1 confirmation 1108:1119 Identity confirmation 1099:1119 Identity confirmation 1099:1119 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 34 theme O-glycans 1150:1158 arg1 elucidation 1135:1145 structure elucidation 1125:1145 structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions 1125:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 8 35 theme O-glycans 1088:1096 arg1 release 1077:1083 quantitative nonselective release 1051:1083 quantitative nonselective release of O-glycans 1051:1096 It was shown that this procedure is superior for quantitative nonselective release of O-glycans. 24161615 9 36 from confirmation 1108:1119 arg1 fractions 1212:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 7 37 theme method 956:961 arg1 parameters 975:984 The method performance parameters 952:984 The method performance parameters 952:984 The method performance parameters were evaluated. 24161615 4 38 theme β-elimination 731:743 arg1 reagent 745:751 ammonia-based β-elimination reagent 717:751 ammonia-based β-elimination reagent 717:751 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 0 39 theme quantitative 33:44 arg1 analysis 62:69 quantitative O-linked glycan analysis 33:69 quantitative O-linked glycan analysis 33:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 9 40 theme ion 1257:1259 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 40 theme ion 1257:1259 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 4 41 dep efficient 678:686 arg1 efficient 678:686 efficient 678:686 efficient 678:686 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 41 dep efficient 678:686 arg1 times 667:671 approximately 20 to 30 times 644:671 approximately 20 to 30 times 644:671 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 41 dep efficient 678:686 arg1 release 600:606 the release 596:606 the release of O-glycans from glycoproteins 596:638 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 1 42 dep optimization 176:187 arg1 the 172:174 the 172:174 the 172:174 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 7 43 theme performance 963:973 arg1 parameters 975:984 The method performance parameters 952:984 The method performance parameters 952:984 The method performance parameters were evaluated. 24161615 1 44 theme quantitative 208:219 arg1 analysis 238:245 quantitative and nonselective analysis 208:245 quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins 208:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 4 45 from glycoproteins 626:638 arg1 efficient 678:686 efficient 678:686 efficient 678:686 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 45 from glycoproteins 626:638 arg1 times 667:671 approximately 20 to 30 times 644:671 approximately 20 to 30 times 644:671 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 45 from glycoproteins 626:638 arg1 release 600:606 the release 596:606 the release of O-glycans from glycoproteins 596:638 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 5 46 theme particular 829:838 arg1 species 847:853 particular glycan species 829:853 particular glycan species 829:853 In addition, the ammonia-based reagent demonstrated bias in the release of particular glycan species. 24161615 9 47 theme LTQ 1303:1305 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 47 theme LTQ 1303:1305 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 0 48 theme glycan 55:60 arg1 analysis 62:69 quantitative O-linked glycan analysis 33:69 quantitative O-linked glycan analysis 33:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 4 49 theme O-glycans 611:619 arg1 efficient 678:686 efficient 678:686 efficient 678:686 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 49 theme O-glycans 611:619 arg1 times 667:671 approximately 20 to 30 times 644:671 approximately 20 to 30 times 644:671 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 4 49 theme O-glycans 611:619 arg1 release 600:606 the release 596:606 the release of O-glycans from glycoproteins 596:638 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 0 50 used used 71:74 arg2 Comparison 0:9 Comparison 0:9 Comparison of two approaches for quantitative O-linked glycan analysis 0:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 1 51 gly glycoproteins 293:305 arg1 glycoproteins 293:305 therapeutic glycoproteins 281:305 therapeutic glycoproteins 281:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 0 52 theme O-linked 46:53 arg1 analysis 62:69 quantitative O-linked glycan analysis 33:69 quantitative O-linked glycan analysis 33:69 Comparison of two approaches for quantitative O-linked glycan analysis used in characterization of recombinant proteins. 24161615 3 53 theme hydrazinolysis 555:568 arg1 procedures 570:579 the ammonia-based and hydrazinolysis procedures 533:579 procedures 570:579 A significant discrepancy in deglycosylation activity was observed between the ammonia-based and hydrazinolysis procedures. 24161615 9 54 theme hydrophilic 1165:1175 arg1 HILIC 1205:1209 HILIC 1205:1209 HILIC 1205:1209 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 54 theme hydrophilic 1165:1175 arg1 chromatography 1189:1202 hydrophilic interaction chromatography 1165:1202 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 6 55 theme quantitative 865:876 arg1 procedure 893:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure was developed for characterization of O-glycans. 24161615 1 56 theme principal 125:133 arg1 aim 135:137 The principal aim 121:137 The principal aim of this study 121:151 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 8 57 theme nonselective 1064:1075 arg1 release 1077:1083 quantitative nonselective release 1051:1083 quantitative nonselective release of O-glycans 1051:1096 It was shown that this procedure is superior for quantitative nonselective release of O-glycans. 24161615 1 58 attach released 267:274 arg1 glycoproteins 293:305 therapeutic glycoproteins 281:305 therapeutic glycoproteins 281:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 1 58 attach released 267:274 arg2 glycans 259:265 O-linked glycans 250:265 O-linked glycans released from therapeutic glycoproteins 250:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 6 59 theme robust 858:863 arg1 procedure 893:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure 856:901 A robust quantitative hydrazinolysis procedure was developed for characterization of O-glycans. 24161615 2 60 theme glycans 360:366 arg1 release 340:346 quantitative release 327:346 quantitative release of O-linked glycans 327:366 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 1 61 theme nonselective 225:236 arg1 analysis 238:245 quantitative and nonselective analysis 208:245 quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins 208:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 4 62 with efficient 678:686 arg1 hydrazine 693:701 hydrazine 693:701 hydrazine 693:701 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 9 63 theme FT 1307:1308 arg1 MS 1310:1311 LTQ FT MS 1303:1311 LTQ FT MS 1303:1311 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 63 theme FT 1307:1308 arg1 spectrometry 1289:1300 linear ion trap Fourier transform mass spectrometry 1250:1300 linear ion trap Fourier transform mass spectrometry (LTQ FT MS) 1250:1312 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 2 64 theme β-elimination 397:409 arg1 strategies 446:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 ammonia-based β-elimination and hydrazinolysis deglycosylation strategies 383:455 Two approaches for quantitative release of O-linked glycans were examined: ammonia-based β-elimination and hydrazinolysis deglycosylation strategies. 24161615 1 65 theme analysis 238:245 arg1 optimization 176:187 optimization 176:187 optimization 176:187 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 1 65 theme analysis 238:245 arg1 fine-tuning 193:203 fine-tuning 193:203 fine-tuning 193:203 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 9 66 theme mass 1319:1322 arg1 accuracy 1324:1331 mass accuracy 1319:1331 mass accuracy below 1ppm 1319:1342 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 4 67 theme ammonia-based 717:729 arg1 reagent 745:751 ammonia-based β-elimination reagent 717:751 ammonia-based β-elimination reagent 717:751 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 9 68 theme interaction 1177:1187 arg1 HILIC 1205:1209 HILIC 1205:1209 HILIC 1205:1209 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 9 68 theme interaction 1177:1187 arg1 chromatography 1189:1202 hydrophilic interaction chromatography 1165:1202 hydrophilic interaction chromatography (HILIC) fractions 1165:1220 Identity confirmation and structure elucidation of O-glycans from hydrophilic interaction chromatography (HILIC) fractions was also demonstrated using linear ion trap Fourier transform mass spectrometry (LTQ FT MS) with mass accuracy below 1ppm. 24161615 4 69 gly glycoproteins 626:638 arg1 glycoproteins 626:638 glycoproteins 626:638 glycoproteins 626:638 Specifically, the release of O-glycans from glycoproteins was approximately 20 to 30 times more efficient with hydrazine compared with ammonia-based β-elimination reagent. 24161615 1 70 theme study 147:151 arg1 aim 135:137 The principal aim 121:137 The principal aim of this study 121:151 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 24161615 1 71 theme O-linked 250:257 arg1 glycans 259:265 O-linked glycans 250:265 O-linked glycans released from therapeutic glycoproteins 250:305 The principal aim of this study was to demonstrate the optimization and fine-tuning of quantitative and nonselective analysis of O-linked glycans released from therapeutic glycoproteins. 26082235 4 0 dep rituximab 703:711 arg1 antibody 739:746 a therapeutic monoclonal antibody 714:746 rituximab (a therapeutic monoclonal antibody) 703:747 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 6 1 theme activated 1264:1272 arg1 oxazoline 1281:1289 activated glycan oxazoline 1264:1289 activated glycan oxazoline 1264:1289 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 7 2 theme sialylated 1335:1344 arg1 semi-synthesis 1317:1330 A semi-synthesis 1315:1330 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines 1315:1392 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines is also described. 26082235 2 3 from domain 322:327 arg1 presence 256:263 presence 256:263 presence 256:263 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 2 3 from domain 322:327 arg1 structures 274:283 fine structures 269:283 fine structures 269:283 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 3 4 theme favorable 491:499 arg1 status 514:519 a favorable, homogeneous status 489:519 a favorable, homogeneous status in various host expression systems 489:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 8 5 theme various 1540:1546 arg1 applications 1548:1559 various applications 1540:1559 various applications 1540:1559 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 5 6 dep endoglycosidase 825:839 arg1 such 842:845 such 842:845 such 842:845 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 3 7 theme various 524:530 arg1 systems 548:554 various host expression systems 524:554 various host expression systems 524:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 6 8 theme GlcNAc 1136:1141 arg1 acceptor 1143:1150 the GlcNAc acceptor 1132:1150 the GlcNAc acceptor 1132:1150 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 8 9 theme homogeneous 1510:1520 arg1 glycoforms 1525:1534 homogeneous Fc glycoforms 1510:1534 homogeneous Fc glycoforms 1510:1534 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 5 10 theme innermost 926:934 arg1 GlcNAc 936:941 only the innermost GlcNAc 917:941 only the innermost GlcNAc 917:941 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 3 11 theme host 532:535 arg1 systems 548:554 various host expression systems 524:554 various host expression systems 524:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 4 12 theme detailed 616:623 arg1 procedure 625:633 a detailed procedure 614:633 a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies 614:694 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 4 13 theme antibodies 685:694 arg1 glyco-engineering 653:669 chemoenzymatic glyco-engineering 638:669 chemoenzymatic glyco-engineering of monoclonal antibodies 638:694 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 3 14 theme glycosylation 472:484 arg1 task 579:582 still a challenging task 559:582 still a challenging task 559:582 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 3 14 theme glycosylation 472:484 arg1 control 461:467 the control 457:467 the control of glycosylation to a favorable, homogeneous status in various host expression systems 457:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 7 15 theme N-glycan 1374:1381 arg1 oxazolines 1383:1392 biantennary N-glycan oxazolines 1362:1392 biantennary N-glycan oxazolines 1362:1392 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines is also described. 26082235 6 16 theme deglycosylated 1010:1023 arg1 IgG 1025:1027 Then the deglycosylated IgG 1001:1027 Then the deglycosylated IgG 1001:1027 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 16 theme deglycosylated 1010:1023 arg1 acceptor 1042:1049 an acceptor 1039:1049 an acceptor for an endoglycosidase-catalyzed transglycosylation 1039:1101 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 2 17 theme antibodies 377:386 arg1 functions 364:372 the effector functions 351:372 the effector functions of antibodies 351:386 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 1 18 theme cancer 173:178 arg1 treatment 160:168 the treatment 156:168 the treatment of cancer, inflammation, and infectious diseases 156:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 6 19 dep enzyme 1253:1258 arg1 the 1249:1251 the 1249:1251 the 1249:1251 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 20 theme glycosynthase 1225:1237 arg1 mutant 1239:1244 a glycosynthase mutant 1223:1244 a glycosynthase mutant 1223:1244 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 20 theme glycosynthase 1225:1237 arg1 substrate 1304:1312 the donor substrate 1294:1312 the donor substrate 1294:1312 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 2 21 theme fine 269:272 arg1 structures 274:283 fine structures 269:283 fine structures 269:283 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 6 22 gly glycoform 1199:1207 arg1 IgG 1212:1214 IgG 1212:1214 IgG 1212:1214 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 5 23 gly glycosylation 981:993 arg2 site 995:998 the glycosylation site 977:998 the glycosylation site 977:998 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 8 24 theme Fc 1522:1523 arg1 glycoforms 1525:1534 homogeneous Fc glycoforms 1510:1534 homogeneous Fc glycoforms 1510:1534 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 8 25 gly glycoforms 1525:1534 arg1 Fc 1522:1523 homogeneous Fc glycoforms 1510:1534 homogeneous Fc glycoforms 1510:1534 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 4 26 theme monoclonal 728:737 arg1 antibody 739:746 a therapeutic monoclonal antibody 714:746 rituximab (a therapeutic monoclonal antibody) 703:747 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 1 27 theme inflammation 181:192 arg1 treatment 160:168 the treatment 156:168 the treatment of cancer, inflammation, and infectious diseases 156:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 0 28 theme Chemoenzymatic 0:13 arg1 Glyco-engineering 15:31 Chemoenzymatic Glyco-engineering 0:31 Chemoenzymatic Glyco-engineering of Monoclonal Antibodies 0:56 Chemoenzymatic Glyco-engineering of Monoclonal Antibodies. 26082235 8 29 used used 1444:1447 arg2 procedure 1427:1435 This detailed procedure 1413:1435 This detailed procedure 1413:1435 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 8 30 theme glycosylation 1460:1472 arg1 remodeling 1474:1483 the Fc glycosylation remodeling 1453:1483 the Fc glycosylation remodeling of other mAbs 1453:1497 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 7 31 dep sialylated 1335:1344 arg1 oxazolines 1383:1392 biantennary N-glycan oxazolines 1362:1392 biantennary N-glycan oxazolines 1362:1392 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines is also described. 26082235 1 32 theme important 95:103 arg1 class 105:109 an important class 92:109 an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases 92:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 1 32 theme important 95:103 arg1 antibodies 70:79 Monoclonal antibodies 59:79 Monoclonal antibodies (mAbs) 59:86 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 2 33 dep presence 256:263 arg1 the 252:254 the 252:254 the 252:254 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 0 34 theme Monoclonal 36:45 arg1 Antibodies 47:56 Monoclonal Antibodies 36:56 Monoclonal Antibodies 36:56 Chemoenzymatic Glyco-engineering of Monoclonal Antibodies. 26082235 4 35 theme therapeutic 716:726 arg1 antibody 739:746 a therapeutic monoclonal antibody 714:746 rituximab (a therapeutic monoclonal antibody) 703:747 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 1 36 gly glycoproteins 126:138 arg1 glycoproteins 126:138 therapeutic glycoproteins 114:138 therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases 114:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 3 37 theme challenging 567:577 arg1 task 579:582 still a challenging task 559:582 still a challenging task 559:582 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 3 37 theme challenging 567:577 arg1 control 461:467 the control 457:467 the control of glycosylation to a favorable, homogeneous status in various host expression systems 457:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 2 38 theme N-glycans 302:310 arg1 presence 256:263 presence 256:263 presence 256:263 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 2 38 theme N-glycans 302:310 arg1 structures 274:283 fine structures 269:283 fine structures 269:283 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 7 39 theme biantennary 1362:1372 arg1 oxazolines 1383:1392 biantennary N-glycan oxazolines 1362:1392 biantennary N-glycan oxazolines 1362:1392 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines is also described. 26082235 5 40 theme mAb 815:817 arg1 deglycosylation 794:808 the deglycosylation 790:808 the deglycosylation of a mAb 790:817 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 3 41 dep favorable 491:499 arg1 homogeneous 502:512 homogeneous 502:512 homogeneous 502:512 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 2 42 theme conserved 292:300 arg1 N-glycans 302:310 the conserved N-glycans 288:310 the conserved N-glycans 288:310 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 1 43 theme infectious 199:208 arg1 diseases 210:217 infectious diseases 199:217 infectious diseases 199:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 5 44 gly core-fucosylated 950:965 arg1 GlcNAc 967:972 the core-fucosylated GlcNAc 946:972 the core-fucosylated GlcNAc 946:972 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 4 45 theme glyco-engineering 653:669 arg1 procedure 625:633 a detailed procedure 614:633 a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies 614:694 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 6 46 theme donor 1298:1302 arg1 mutant 1239:1244 a glycosynthase mutant 1223:1244 a glycosynthase mutant 1223:1244 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 46 theme donor 1298:1302 arg1 substrate 1304:1312 the donor substrate 1294:1312 the donor substrate 1294:1312 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 3 47 theme Fc 439:440 arg1 glycoforms 442:451 Fc glycoforms 439:451 Fc glycoforms 439:451 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 5 48 gly deglycosylation 794:808 arg1 mAb 815:817 a mAb 813:817 a mAb 813:817 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 4 49 theme chemoenzymatic 638:651 arg1 glyco-engineering 653:669 chemoenzymatic glyco-engineering 638:669 chemoenzymatic glyco-engineering of monoclonal antibodies 638:694 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 1 50 theme diseases 210:217 arg1 treatment 160:168 the treatment 156:168 the treatment of cancer, inflammation, and infectious diseases 156:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 0 51 theme Antibodies 47:56 arg1 Glyco-engineering 15:31 Chemoenzymatic Glyco-engineering 0:31 Chemoenzymatic Glyco-engineering of Monoclonal Antibodies 0:56 Chemoenzymatic Glyco-engineering of Monoclonal Antibodies. 26082235 5 52 theme glycosylation 981:993 arg1 site 995:998 the glycosylation site 977:998 the glycosylation site 977:998 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 7 53 theme asialylated 1350:1360 arg1 semi-synthesis 1317:1330 A semi-synthesis 1315:1330 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines 1315:1392 A semi-synthesis of sialylated and asialylated biantennary N-glycan oxazolines is also described. 26082235 3 54 theme glycoforms 442:451 arg1 mixtures 427:434 mixtures 427:434 mixtures of Fc glycoforms 427:451 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 3 54 theme glycoforms 442:451 arg1 mAbs 398:401 mAbs 398:401 mAbs 398:401 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 5 55 theme type 855:858 arg1 EndoS 860:864 wild type EndoS 850:864 wild type EndoS 850:864 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 3 56 gly glycoforms 442:451 arg1 Fc 439:440 Fc glycoforms 439:451 Fc glycoforms 439:451 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 1 57 theme therapeutic 114:124 arg1 glycoproteins 126:138 therapeutic glycoproteins 114:138 therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases 114:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 8 58 theme mAbs 1494:1497 arg1 remodeling 1474:1483 the Fc glycosylation remodeling 1453:1483 the Fc glycosylation remodeling of other mAbs 1453:1497 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 6 59 gly deglycosylated 1010:1023 arg1 IgG 1025:1027 Then the deglycosylated IgG 1001:1027 Then the deglycosylated IgG 1001:1027 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 59 gly deglycosylated 1010:1023 arg1 acceptor 1042:1049 an acceptor 1039:1049 an acceptor for an endoglycosidase-catalyzed transglycosylation 1039:1101 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 8 60 theme Fc 1457:1458 arg1 remodeling 1474:1483 the Fc glycosylation remodeling 1453:1483 the Fc glycosylation remodeling of other mAbs 1453:1497 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 2 61 theme Compelling 220:229 arg1 data 231:234 Compelling data 220:234 Compelling data 220:234 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 1 62 theme glycoproteins 126:138 arg1 class 105:109 an important class 92:109 an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases 92:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 1 62 theme glycoproteins 126:138 arg1 antibodies 70:79 Monoclonal antibodies 59:79 Monoclonal antibodies (mAbs) 59:86 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 8 63 theme detailed 1418:1425 arg1 procedure 1427:1435 This detailed procedure 1413:1435 This detailed procedure 1413:1435 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 6 64 theme glycan 1274:1279 arg1 oxazoline 1281:1289 activated glycan oxazoline 1264:1289 activated glycan oxazoline 1264:1289 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 5 65 theme wild 850:853 arg1 EndoS 860:864 wild type EndoS 850:864 wild type EndoS 850:864 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 6 66 theme natural 1191:1197 arg1 glycoform 1199:1207 a defined, homogeneous natural glycoform 1168:1207 a defined, homogeneous natural glycoform of IgG 1168:1214 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 6 67 theme IgG 1212:1214 arg1 glycoform 1199:1207 a defined, homogeneous natural glycoform 1168:1207 a defined, homogeneous natural glycoform of IgG 1168:1214 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 5 68 theme heterogeneous 881:893 arg1 N-glycans 898:906 the heterogeneous Fc N-glycans 877:906 the heterogeneous Fc N-glycans 877:906 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 3 69 theme expression 537:546 arg1 systems 548:554 various host expression systems 524:554 various host expression systems 524:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 26082235 2 70 attach presence 256:263 arg2 N-glycans 302:310 the conserved N-glycans 288:310 the conserved N-glycans 288:310 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 2 70 attach presence 256:263 arg1 domain 322:327 the Fc domain 315:327 the Fc domain 315:327 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 4 71 theme model 754:758 arg1 rituximab 703:711 rituximab 703:711 rituximab (a therapeutic monoclonal antibody) 703:747 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 4 71 theme model 754:758 arg1 system 760:765 a model system 752:765 a model system 752:765 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 5 72 theme core-fucosylated 950:965 arg1 GlcNAc 967:972 the core-fucosylated GlcNAc 946:972 the core-fucosylated GlcNAc 946:972 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 2 73 theme Fc 319:320 arg1 domain 322:327 the Fc domain 315:327 the Fc domain 315:327 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 6 74 theme desired 1112:1118 arg1 N-glycan 1120:1127 a desired N-glycan 1110:1127 a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG 1110:1214 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 5 75 theme Fc 895:896 arg1 N-glycans 898:906 the heterogeneous Fc N-glycans 877:906 the heterogeneous Fc N-glycans 877:906 The protocol includes the deglycosylation of a mAb by an endoglycosidase (such as wild type EndoS) to remove the heterogeneous Fc N-glycans, leaving only the innermost GlcNAc or the core-fucosylated GlcNAc at the glycosylation site. 26082235 6 76 theme endoglycosidase-catalyzed 1058:1082 arg1 transglycosylation 1084:1101 an endoglycosidase-catalyzed transglycosylation 1055:1101 an endoglycosidase-catalyzed transglycosylation 1055:1101 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 1 77 theme Monoclonal 59:68 arg1 class 105:109 an important class 92:109 an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases 92:217 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 1 77 theme Monoclonal 59:68 arg1 mAbs 82:85 mAbs 82:85 mAbs 82:85 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 1 77 theme Monoclonal 59:68 arg1 antibodies 70:79 Monoclonal antibodies 59:79 Monoclonal antibodies (mAbs) 59:86 Monoclonal antibodies (mAbs) are an important class of therapeutic glycoproteins widely used for the treatment of cancer, inflammation, and infectious diseases. 26082235 4 78 theme monoclonal 674:683 arg1 antibodies 685:694 monoclonal antibodies 674:694 monoclonal antibodies 674:694 In this chapter, we describe a detailed procedure of chemoenzymatic glyco-engineering of monoclonal antibodies, using rituximab (a therapeutic monoclonal antibody) as a model system. 26082235 6 79 dep defined 1170:1176 arg1 homogeneous 1179:1189 homogeneous 1179:1189 homogeneous 1179:1189 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 8 80 theme other 1488:1492 arg1 mAbs 1494:1497 other mAbs 1488:1497 other mAbs 1488:1497 This detailed procedure can be used for the Fc glycosylation remodeling of other mAbs to provide homogeneous Fc glycoforms for various applications. 26082235 6 81 theme defined 1170:1176 arg1 glycoform 1199:1207 a defined, homogeneous natural glycoform 1168:1207 a defined, homogeneous natural glycoform of IgG 1168:1214 Then the deglycosylated IgG serves as an acceptor for an endoglycosidase-catalyzed transglycosylation to add a desired N-glycan to the GlcNAc acceptor to reconstitute a defined, homogeneous natural glycoform of IgG, using a glycosynthase mutant as the enzyme and activated glycan oxazoline as the donor substrate. 26082235 2 82 theme effector 355:362 arg1 functions 364:372 the effector functions 351:372 the effector functions of antibodies 351:386 Compelling data have shown that the presence and fine structures of the conserved N-glycans at the Fc domain can profoundly affect the effector functions of antibodies. 26082235 3 83 from status 514:519 arg1 systems 548:554 various host expression systems 524:554 various host expression systems 524:554 However, mAbs are usually produced as mixtures of Fc glycoforms and the control of glycosylation to a favorable, homogeneous status in various host expression systems is still a challenging task. 25213144 0 0 theme allelic 80:86 arg1 polymorphisms 88:100 allelic polymorphisms 80:100 allelic polymorphisms 80:100 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 0 1 from diversity 16:24 arg1 system 63:68 a bacterial protein glycosylation system 29:68 a bacterial protein glycosylation system 29:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 1 2 link O-linked 221:228 arg1 glycosylation 238:250 O-linked protein glycosylation 221:250 O-linked protein glycosylation in species within the genus Neisseria display intra- 221:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 8 3 theme polysaccharide 1456:1469 arg1 variability 1471:1481 capsular polysaccharide variability 1447:1481 capsular polysaccharide variability 1447:1481 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 7 4 theme N. meningitidis 1292:1306 arg1 strains 1281:1287 strains 1281:1287 strains of N. meningitidis 1281:1306 Phylogenetic analyses revealed that GlcNAc-containing disaccharides may be widely distributed within the pgl systems of Neisseria particularly in strains of N. meningitidis. 25213144 3 5 from genes 542:546 arg1 degree 492:497 the high degree 483:497 the high degree of standing allelic polymorphisms in these genes 483:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 7 6 theme GlcNAc-containing 1171:1187 arg1 disaccharides 1189:1201 GlcNAc-containing disaccharides 1171:1201 GlcNAc-containing disaccharides 1171:1201 Phylogenetic analyses revealed that GlcNAc-containing disaccharides may be widely distributed within the pgl systems of Neisseria particularly in strains of N. meningitidis. 25213144 8 7 theme minimal 1328:1334 arg1 alterations 1347:1357 analogous minimal structural alterations 1318:1357 analogous minimal structural alterations in glycosyltransferases 1318:1381 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 1 8 theme genus 274:278 arg1 intra- 298:303 the genus Neisseria display intra- 270:303 the genus Neisseria display intra- 270:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 4 9 theme protein-associated 639:656 arg1 glycoforms 671:680 unique protein-associated disaccharide glycoforms 632:680 unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end 632:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 1 10 gly glycosylation 238:250 arg1 species 255:261 species 255:261 species within the genus Neisseria display intra- 255:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 3 11 theme diversity 569:577 arg1 level 553:557 the level 549:557 the level of glycan diversity 549:577 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 1 12 theme Neisseria 280:288 arg1 intra- 298:303 the genus Neisseria display intra- 270:303 the genus Neisseria display intra- 270:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 6 13 theme site-specific 962:974 arg1 mutagenesis 976:986 site-specific mutagenesis 962:986 site-specific mutagenesis 962:986 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 3 14 from degree 492:497 arg1 genes 542:546 these genes 536:546 these genes 536:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 1 15 theme display 290:296 arg1 intra- 298:303 the genus Neisseria display intra- 270:303 the genus Neisseria display intra- 270:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 0 16 theme genetic 114:120 arg1 alterations 122:132 minimal genetic alterations 106:132 minimal genetic alterations 106:132 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 4 17 gly glycoforms 671:680 arg1 disaccharide 658:669 unique protein-associated disaccharide glycoforms 632:680 unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end 632:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 3 18 theme standing 502:509 arg1 polymorphisms 519:531 standing allelic polymorphisms 502:531 standing allelic polymorphisms in these genes 502:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 2 19 theme expression 432:441 arg1 status 443:448 expression status 432:448 expression status 432:448 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 0 20 theme minimal 106:112 arg1 alterations 122:132 minimal genetic alterations 106:132 minimal genetic alterations 106:132 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 6 21 theme Allele 939:944 arg1 comparisons 946:956 Allele comparisons 939:956 Allele comparisons 939:956 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 3 22 theme glycan 562:567 arg1 diversity 569:577 glycan diversity 562:577 glycan diversity 562:577 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 5 23 theme allelic 792:798 arg1 variants 800:807 allelic variants 792:807 allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides 792:936 This altered structure was correlated with allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides. 25213144 0 24 theme glycan 9:14 arg1 diversity 16:24 Extended glycan diversity 0:24 Extended glycan diversity in a bacterial protein glycosylation system 0:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 3 25 from polymorphisms 519:531 arg1 genes 542:546 these genes 536:546 these genes 536:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 2 26 theme glycan 347:352 arg1 structure 354:362 glycan structure 347:362 glycan structure 347:362 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 5 27 theme -containing 912:922 arg1 disaccharides 924:936 glucose (Glc)-containing disaccharides 899:936 glucose (Glc)-containing disaccharides 899:936 This altered structure was correlated with allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides. 25213144 0 28 theme Extended 0:7 arg1 diversity 16:24 Extended glycan diversity 0:24 Extended glycan diversity in a bacterial protein glycosylation system 0:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 7 29 theme Phylogenetic 1135:1146 arg1 analyses 1148:1155 Phylogenetic analyses 1135:1155 Phylogenetic analyses 1135:1155 Phylogenetic analyses revealed that GlcNAc-containing disaccharides may be widely distributed within the pgl systems of Neisseria particularly in strains of N. meningitidis. 25213144 6 30 from position 1045:1052 arg1 residue 1025:1031 a single residue 1016:1031 a single residue 1016:1031 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 6 30 from position 1045:1052 arg1 alanine 1034:1040 alanine 1034:1040 alanine at position 303 in place of a glutamine 1034:1080 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 5 31 theme disaccharides 924:936 arg1 expression 885:894 the expression 881:894 the expression of glucose (Glc)-containing disaccharides 881:936 This altered structure was correlated with allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides. 25213144 6 32 theme residue 1025:1031 arg1 sufficient 1087:1096 sufficient 1087:1096 sufficient 1087:1096 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 6 32 theme residue 1025:1031 arg1 presence 1004:1011 the presence 1000:1011 the presence of a single residue, alanine at position 303 in place of a glutamine, 1000:1081 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 2 33 from differences 401:411 arg1 content 420:426 content 420:426 content 420:426 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 2 33 from differences 401:411 arg1 status 443:448 expression status 432:448 expression status 432:448 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 8 34 theme first 1507:1511 arg1 this 1484:1487 this 1484:1487 this 1484:1487 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 8 34 theme first 1507:1511 arg1 example 1513:1519 the first example 1503:1519 the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system 1503:1636 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 1 35 theme broad 206:210 arg1 spectrum 212:219 the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity 202:329 spectrum 212:219 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 2 36 theme genes 470:474 arg1 content 420:426 content 420:426 content 420:426 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 2 36 theme genes 470:474 arg1 status 443:448 expression status 432:448 expression status 432:448 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 1 37 theme interstrain 309:319 arg1 diversity 321:329 interstrain diversity 309:329 interstrain diversity 309:329 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 5 38 theme altered 754:760 arg1 structure 762:770 This altered structure 749:770 This altered structure 749:770 This altered structure was correlated with allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides. 25213144 4 39 contain carry 687:691 arg2 GlcNAc 714:719 GlcNAc 714:719 GlcNAc 714:719 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 4 39 contain carry 687:691 arg1 glycoforms 671:680 unique protein-associated disaccharide glycoforms 632:680 unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end 632:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 4 39 contain carry 687:691 arg2 N-acetylglucosamine 693:711 N-acetylglucosamine 693:711 N-acetylglucosamine (GlcNAc) 693:720 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 4 39 contain carry 687:691 arg3 end 744:746 their non-reducing end 725:746 their non-reducing end 725:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 8 40 theme capsular 1447:1454 arg1 polysaccharide 1456:1469 capsular polysaccharide 1447:1469 capsular polysaccharide variability 1447:1481 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 2 41 theme glycan 453:458 arg1 genes 470:474 glycan synthesis genes 453:474 glycan synthesis genes 453:474 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 0 42 theme bacterial 31:39 arg1 system 63:68 a bacterial protein glycosylation system 29:68 a bacterial protein glycosylation system 29:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 6 43 theme single 1018:1023 arg1 residue 1025:1031 a single residue 1016:1031 a single residue 1016:1031 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 6 43 theme single 1018:1023 arg1 alanine 1034:1040 alanine 1034:1040 alanine at position 303 in place of a glutamine 1034:1080 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 0 44 theme glycosyltransferase 139:157 arg1 gene 159:162 a glycosyltransferase gene 137:162 a glycosyltransferase gene 137:162 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 3 45 theme high 487:490 arg1 degree 492:497 the high degree 483:497 the high degree of standing allelic polymorphisms in these genes 483:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 8 46 theme structural 1336:1345 arg1 alterations 1347:1357 analogous minimal structural alterations 1318:1357 analogous minimal structural alterations in glycosyltransferases 1318:1381 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 1 47 theme O-linked 221:228 arg1 glycosylation 238:250 O-linked protein glycosylation 221:250 O-linked protein glycosylation in species within the genus Neisseria display intra- 221:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 8 48 theme analogous 1318:1326 arg1 alterations 1347:1357 analogous minimal structural alterations 1318:1357 analogous minimal structural alterations in glycosyltransferases 1318:1381 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 8 49 theme glycan 1567:1572 arg1 diversification 1574:1588 glycan diversification 1567:1588 glycan diversification within a bacterial protein glycosylation system 1567:1636 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 1 50 theme protein 230:236 arg1 glycosylation 238:250 O-linked protein glycosylation 221:250 O-linked protein glycosylation in species within the genus Neisseria display intra- 221:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 0 51 theme glycosylation 49:61 arg1 system 63:68 a bacterial protein glycosylation system 29:68 a bacterial protein glycosylation system 29:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 8 52 theme such 1530:1533 arg1 changes 1535:1541 such changes 1530:1541 such changes 1530:1541 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 0 53 attach linked 70:75 arg1 alterations 122:132 minimal genetic alterations 106:132 minimal genetic alterations 106:132 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 0 53 attach linked 70:75 arg1 polymorphisms 88:100 allelic polymorphisms 80:100 allelic polymorphisms 80:100 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 0 53 attach linked 70:75 arg2 diversity 16:24 Extended glycan diversity 0:24 Extended glycan diversity in a bacterial protein glycosylation system 0:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 8 54 with association 1407:1417 arg1 variability 1471:1481 capsular polysaccharide variability 1447:1481 capsular polysaccharide variability 1447:1481 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 8 54 with association 1407:1417 arg1 lipopolysaccharide 1424:1441 lipopolysaccharide 1424:1441 lipopolysaccharide 1424:1441 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 0 55 theme protein 41:47 arg1 system 63:68 a bacterial protein glycosylation system 29:68 a bacterial protein glycosylation system 29:68 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 8 56 theme bacterial 1599:1607 arg1 system 1631:1636 a bacterial protein glycosylation system 1597:1636 a bacterial protein glycosylation system 1597:1636 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 1 57 from glycosylation 238:250 arg1 species 255:261 species 255:261 species within the genus Neisseria display intra- 255:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 2 58 dep content 420:426 arg1 the 416:418 the 416:418 the 416:418 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 3 59 theme allelic 511:517 arg1 polymorphisms 519:531 standing allelic polymorphisms 502:531 standing allelic polymorphisms in these genes 502:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 4 60 theme unique 632:637 arg1 glycoforms 671:680 unique protein-associated disaccharide glycoforms 632:680 unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end 632:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 1 61 with conjunction 185:195 arg1 spectrum 212:219 the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity 202:329 spectrum 212:219 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 1 61 with conjunction 185:195 arg1 diversity 321:329 interstrain diversity 309:329 interstrain diversity 309:329 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 8 62 theme glycosylation 1617:1629 arg1 system 1631:1636 a bacterial protein glycosylation system 1597:1636 a bacterial protein glycosylation system 1597:1636 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 8 63 from alterations 1347:1357 arg1 glycosyltransferases 1362:1381 glycosyltransferases 1362:1381 glycosyltransferases 1362:1381 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 3 64 theme polymorphisms 519:531 arg1 degree 492:497 the high degree 483:497 the high degree of standing allelic polymorphisms in these genes 483:546 Given the high degree of standing allelic polymorphisms in these genes, the level of glycan diversity may exceed that currently defined. 25213144 8 65 theme protein 1609:1615 arg1 system 1631:1636 a bacterial protein glycosylation system 1597:1636 a bacterial protein glycosylation system 1597:1636 Although analogous minimal structural alterations in glycosyltransferases have been documented in association with lipopolysaccharide and capsular polysaccharide variability, this appears to be the first example in which such changes have been implicated in glycan diversification within a bacterial protein glycosylation system. 25213144 0 66 from polymorphisms 88:100 arg1 gene 159:162 a glycosyltransferase gene 137:162 a glycosyltransferase gene 137:162 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 25213144 2 67 theme synthesis 460:468 arg1 genes 470:474 glycan synthesis genes 453:474 glycan synthesis genes 453:474 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 4 68 theme non-reducing 731:742 arg1 end 744:746 their non-reducing end 725:746 their non-reducing end 725:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 6 69 theme Glc 1116:1118 arg1 incorporation 1120:1132 Glc incorporation 1116:1132 Glc incorporation 1116:1132 Allele comparisons and site-specific mutagenesis showed that the presence of a single residue, alanine at position 303 in place of a glutamine, was sufficient for GlcNAc versus Glc incorporation. 25213144 4 70 theme disaccharide 658:669 arg1 glycoforms 671:680 unique protein-associated disaccharide glycoforms 632:680 unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end 632:746 Here, we identify unique protein-associated disaccharide glycoforms that carry N-acetylglucosamine (GlcNAc) at their non-reducing end. 25213144 5 71 theme pglH 812:815 arg1 variants 800:807 allelic variants 792:807 allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides 792:936 This altered structure was correlated with allelic variants of pglH whose product was previously demonstrated to be responsible for the expression of glucose (Glc)-containing disaccharides. 25213144 7 72 theme Neisseria 1255:1263 arg1 systems 1244:1250 the pgl systems 1236:1250 the pgl systems of Neisseria 1236:1263 Phylogenetic analyses revealed that GlcNAc-containing disaccharides may be widely distributed within the pgl systems of Neisseria particularly in strains of N. meningitidis. 25213144 1 73 dep spectrum 212:219 arg1 glycosylation 238:250 O-linked protein glycosylation 221:250 O-linked protein glycosylation in species within the genus Neisseria display intra- 221:303 Glycans manifest in conjunction with the broad spectrum O-linked protein glycosylation in species within the genus Neisseria display intra- and interstrain diversity. 25213144 7 74 theme pgl 1240:1242 arg1 systems 1244:1250 the pgl systems 1236:1250 the pgl systems of Neisseria 1236:1263 Phylogenetic analyses revealed that GlcNAc-containing disaccharides may be widely distributed within the pgl systems of Neisseria particularly in strains of N. meningitidis. 25213144 2 75 from Variability 332:342 arg1 antigenicity 368:379 antigenicity 368:379 antigenicity 368:379 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 2 75 from Variability 332:342 arg1 structure 354:362 glycan structure 347:362 glycan structure 347:362 Variability in glycan structure and antigenicity are attributable to differences in the content and expression status of glycan synthesis genes. 25213144 0 76 from alterations 122:132 arg1 gene 159:162 a glycosyltransferase gene 137:162 a glycosyltransferase gene 137:162 Extended glycan diversity in a bacterial protein glycosylation system linked to allelic polymorphisms and minimal genetic alterations in a glycosyltransferase gene. 28460017 0 0 theme glycosylation 79:91 arg1 system 93:98 a bacterial protein glycosylation system 59:98 a bacterial protein glycosylation system 59:98 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 3 1 gly glycosylation 460:472 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 1 gly glycosylation 460:472 arg1 species 498:504 pathogenic Neisseria species 477:504 pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 477:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 1 gly glycosylation 460:472 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 2 from species 498:504 arg1 studies 413:419 Previous studies 404:419 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 404:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 5 3 theme only 940:943 arg1 exception 945:953 the only exception 936:953 the only exception of di-N-acetyl-bacillosamine 936:982 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 0 4 theme protein 71:77 arg1 system 93:98 a bacterial protein glycosylation system 59:98 a bacterial protein glycosylation system 59:98 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 4 5 theme complete 740:747 arg1 set 749:751 a complete set 738:751 a complete set of all glycan variants identified to date 738:793 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 4 5 theme complete 740:747 arg1 variants 767:774 all glycan variants 756:774 all glycan variants identified to date 756:793 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 7 6 theme isogenic 1206:1213 arg1 backgrounds 1215:1225 otherwise isogenic backgrounds 1196:1225 otherwise isogenic backgrounds 1196:1225 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 8 7 theme unprecedented 1248:1260 arg1 phenomenon 1262:1271 this unprecedented phenomenon 1243:1271 this unprecedented phenomenon 1243:1271 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 3 8 from glycosylation 460:472 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 8 from glycosylation 460:472 arg1 species 498:504 pathogenic Neisseria species 477:504 pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 477:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 8 from glycosylation 460:472 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 4 9 theme mass 709:712 arg1 spectrometry 714:725 mass spectrometry 709:725 mass spectrometry 709:725 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 5 10 from glycoforms 827:836 arg1 oligosaccharide 860:874 the oligosaccharide 856:874 the oligosaccharide 856:874 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 7 11 theme otherwise 1196:1204 arg1 backgrounds 1215:1225 otherwise isogenic backgrounds 1196:1225 otherwise isogenic backgrounds 1196:1225 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 3 12 theme diverse 598:604 arg1 glycoforms 606:615 their diverse glycoforms 592:615 their diverse glycoforms 592:615 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 4 13 theme glycan 760:765 arg1 variants 767:774 all glycan variants 756:774 all glycan variants identified to date 756:793 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 3 14 from studies 413:419 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 14 from studies 413:419 arg1 species 498:504 pathogenic Neisseria species 477:504 pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 477:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 14 from studies 413:419 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 7 15 theme oligosaccharides 1166:1181 arg1 lengths 1155:1161 altered chain lengths 1141:1161 altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds 1141:1225 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 2 16 theme important 300:308 arg1 consequences 310:321 important consequences 300:321 important consequences for biotic and abiotic recognition events 300:363 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 5 17 theme PglI-dependent 908:921 arg1 fashion 923:929 a PglI-dependent fashion 906:929 a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine 906:982 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 3 18 theme Neisseria 488:496 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 18 theme Neisseria 488:496 arg1 species 498:504 pathogenic Neisseria species 477:504 pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 477:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 18 theme Neisseria 488:496 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 19 theme O-linked 443:450 arg1 glycosylation 460:472 the broad-spectrum O-linked protein glycosylation 424:472 the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 424:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 5 20 from sugars 846:851 arg1 oligosaccharide 860:874 the oligosaccharide 856:874 the oligosaccharide 856:874 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 3 21 theme protein 452:458 arg1 glycosylation 460:472 the broad-spectrum O-linked protein glycosylation 424:472 the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 424:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 7 22 theme chain 1149:1153 arg1 lengths 1155:1161 altered chain lengths 1141:1161 altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds 1141:1225 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 8 23 theme membrane 1359:1366 arg1 topology 1368:1375 the membrane topology 1355:1375 the membrane topology of glycan O-acetylation 1355:1399 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 2 24 from fitness 384:390 arg1 general 395:401 general 395:401 general 395:401 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 2 25 theme recognition 346:356 arg1 events 358:363 biotic and abiotic recognition events 327:363 biotic and abiotic recognition events 327:363 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 3 26 theme some 584:587 arg1 O-acetylation 567:579 O-acetylation 567:579 O-acetylation of some of their diverse glycoforms 567:615 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 1 27 theme common 178:183 arg1 modification 185:196 a common modification 176:196 a common modification of bacterial glycoconjugates 176:225 O-acetylation is a common modification of bacterial glycoconjugates. 28460017 1 27 theme common 178:183 arg1 O-acetylation 159:171 O-acetylation 159:171 O-acetylation 159:171 O-acetylation is a common modification of bacterial glycoconjugates. 28460017 0 28 theme Structural 0:9 arg1 analyses 23:30 Structural and genetic analyses 0:30 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 0:157 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 2 29 contain has 296:298 arg2 consequences 310:321 important consequences 300:321 important consequences for biotic and abiotic recognition events 300:363 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 2 29 contain has 296:298 arg1 O-acetylation 282:294 O-acetylation 282:294 O-acetylation 282:294 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 2 30 theme abiotic 338:344 arg1 events 358:363 biotic and abiotic recognition events 327:363 biotic and abiotic recognition events 327:363 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 3 31 theme pathogenic 477:486 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 31 theme pathogenic 477:486 arg1 species 498:504 pathogenic Neisseria species 477:504 pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 477:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 31 theme pathogenic 477:486 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 3 32 theme Previous 404:411 arg1 studies 413:419 Previous studies 404:419 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 404:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 7 33 theme altered 1141:1147 arg1 lengths 1155:1161 altered chain lengths 1141:1161 altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds 1141:1225 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 3 34 theme N. 517:518 arg1 gonorrhoeae 520:530 N. gonorrhoeae 517:530 N. gonorrhoeae 517:530 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 0 35 theme genetic 15:21 arg1 analyses 23:30 Structural and genetic analyses 0:30 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 0:157 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 6 36 theme single 1016:1021 arg1 oligosaccharide 1023:1037 a single oligosaccharide 1014:1037 a single oligosaccharide 1014:1037 Moreover, multiple sugars in a single oligosaccharide could be simultaneously modified. 28460017 0 37 theme differential 114:125 arg1 effects 127:133 differential effects 114:133 differential effects on glycan chain length 114:156 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 2 38 theme biotic 327:332 arg1 events 358:363 biotic and abiotic recognition events 327:363 biotic and abiotic recognition events 327:363 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 1 39 theme bacterial 201:209 arg1 glycoconjugates 211:225 bacterial glycoconjugates 201:225 bacterial glycoconjugates 201:225 O-acetylation is a common modification of bacterial glycoconjugates. 28460017 0 40 theme glycan 35:40 arg1 O-acetylation 42:54 glycan O-acetylation 35:54 glycan O-acetylation 35:54 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 5 41 with fashion 923:929 arg1 exception 945:953 the only exception 936:953 the only exception of di-N-acetyl-bacillosamine 936:982 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 0 42 theme glycan 138:143 arg1 length 151:156 glycan chain length 138:156 glycan chain length 138:156 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 8 43 theme O-acetylation 1387:1399 arg1 topology 1368:1375 the membrane topology 1355:1375 the membrane topology of glycan O-acetylation 1355:1399 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 1 44 theme glycoconjugates 211:225 arg1 modification 185:196 a common modification 176:196 a common modification of bacterial glycoconjugates 176:225 O-acetylation is a common modification of bacterial glycoconjugates. 28460017 1 44 theme glycoconjugates 211:225 arg1 O-acetylation 159:171 O-acetylation 159:171 O-acetylation 159:171 O-acetylation is a common modification of bacterial glycoconjugates. 28460017 3 45 theme glycosylation 460:472 arg1 studies 413:419 Previous studies 404:419 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 404:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 2 46 theme oligosaccharide 241:255 arg1 structure 257:265 oligosaccharide structure 241:265 oligosaccharide structure 241:265 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 7 47 theme O-acetylation 1088:1100 arg1 status 1102:1107 O-acetylation status 1088:1107 O-acetylation status 1088:1107 Interestingly, O-acetylation status was found to be correlated with altered chain lengths of oligosaccharides expressed in otherwise isogenic backgrounds. 28460017 8 48 theme glycan 1380:1385 arg1 O-acetylation 1387:1399 glycan O-acetylation 1380:1399 glycan O-acetylation 1380:1399 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 3 49 theme broad-spectrum 428:441 arg1 glycosylation 460:472 the broad-spectrum O-linked protein glycosylation 424:472 the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 424:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 5 50 from acetylation 891:901 arg1 fashion 923:929 a PglI-dependent fashion 906:929 a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine 906:982 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 2 51 theme bacterial 374:382 arg1 fitness 384:390 bacterial fitness 374:390 bacterial fitness in general 374:401 By modifying oligosaccharide structure and chemistry, O-acetylation has important consequences for biotic and abiotic recognition events and thus bacterial fitness in general. 28460017 0 52 theme O-acetylation 42:54 arg1 analyses 23:30 Structural and genetic analyses 0:30 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 0:157 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 6 53 theme multiple 995:1002 arg1 sugars 1004:1009 multiple sugars 995:1009 multiple sugars in a single oligosaccharide 995:1037 Moreover, multiple sugars in a single oligosaccharide could be simultaneously modified. 28460017 0 54 theme chain 145:149 arg1 length 151:156 glycan chain length 138:156 glycan chain length 138:156 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 8 55 contain having 1309:1314 arg2 implications 1338:1349 potentially important implications 1316:1349 potentially important implications for the membrane topology of glycan O-acetylation 1316:1399 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 8 55 contain having 1309:1314 arg1 some 1304:1307 some 1304:1307 some 1304:1307 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 5 56 theme di-N-acetyl-bacillosamine 958:982 arg1 exception 945:953 the only exception 936:953 the only exception of di-N-acetyl-bacillosamine 936:982 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 9 57 theme better 1433:1438 arg1 insight 1440:1446 better insight 1433:1446 better insight into how O-acetylation can both directly and indirectly govern glycoform structure and diversity 1433:1543 Together, the findings provide better insight into how O-acetylation can both directly and indirectly govern glycoform structure and diversity. 28460017 5 58 gly glycoforms 827:836 arg2 oligosaccharide 860:874 the oligosaccharide 856:874 the oligosaccharide 856:874 Regardless of composition, all glycoforms and all sugars in the oligosaccharide are subject to acetylation in a PglI-dependent fashion with the only exception of di-N-acetyl-bacillosamine. 28460017 0 59 dep analyses 23:30 arg1 evidence 101:108 evidence 101:108 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 0:157 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 3 60 link O-linked 443:450 arg1 glycosylation 460:472 the broad-spectrum O-linked protein glycosylation 424:472 the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) 424:551 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 4 61 theme variants 767:774 arg1 set 749:751 a complete set 738:751 a complete set of all glycan variants identified to date 738:793 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 4 61 theme variants 767:774 arg1 variants 767:774 all glycan variants 756:774 all glycan variants identified to date 756:793 Herein, we extend these observations by using mass spectrometry to examine a complete set of all glycan variants identified to date. 28460017 6 62 from sugars 1004:1009 arg1 oligosaccharide 1023:1037 a single oligosaccharide 1014:1037 a single oligosaccharide 1014:1037 Moreover, multiple sugars in a single oligosaccharide could be simultaneously modified. 28460017 3 63 theme N. 536:537 arg1 meningitidis 539:550 N. meningitidis 536:550 N. meningitidis 536:550 Previous studies of the broad-spectrum O-linked protein glycosylation in pathogenic Neisseria species (including N. gonorrhoeae and N. meningitidis) have revealed O-acetylation of some of their diverse glycoforms and identified the committed acetylase, PglI. 28460017 9 64 theme glycoform 1511:1519 arg1 structure 1521:1529 glycoform structure 1511:1529 glycoform structure 1511:1529 Together, the findings provide better insight into how O-acetylation can both directly and indirectly govern glycoform structure and diversity. 28460017 0 65 theme bacterial 61:69 arg1 system 93:98 a bacterial protein glycosylation system 59:98 a bacterial protein glycosylation system 59:98 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 0 66 from effects 127:133 arg1 length 151:156 glycan chain length 138:156 glycan chain length 138:156 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28460017 8 67 theme important 1328:1336 arg1 implications 1338:1349 potentially important implications 1316:1349 potentially important implications for the membrane topology of glycan O-acetylation 1316:1399 Models for how this unprecedented phenomenon might arise are discussed with some having potentially important implications for the membrane topology of glycan O-acetylation. 28460017 0 68 from analyses 23:30 arg1 system 93:98 a bacterial protein glycosylation system 59:98 a bacterial protein glycosylation system 59:98 Structural and genetic analyses of glycan O-acetylation in a bacterial protein glycosylation system: evidence for differential effects on glycan chain length. 28091715 5 0 theme O-glycans 848:856 arg1 release 825:831 efficient release 815:831 efficient release of the reduced O-glycans from the glycoproteins 815:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 4 1 theme peptide 643:649 arg1 F 665:665 peptide N-glycosidase F 643:665 peptide N-glycosidase F 643:665 In contrast, N-glycans can be readily released non-reductively from peptide backbones using an enzyme such as peptide N-glycosidase F. 28091715 6 2 theme cytotoxic 899:907 arg1 models 994:999 models 994:999 models for O-glycan analysis 994:1021 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 2 theme cytotoxic 899:907 arg1 lymphocyte 911:920 the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin 895:989 the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin 895:989 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 3 theme 4-immunoglobulin 941:956 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 4 theme 2-AB 1081:1084 arg1 labelling 1086:1094 2-AB labelling 1081:1094 2-AB labelling 1081:1094 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 4 theme 2-AB 1081:1084 arg1 methods 1051:1057 the analytical methods 1036:1057 the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling 1036:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 3 5 theme protein 415:421 arg1 glycosylation 423:435 protein glycosylation 415:435 protein glycosylation 415:435 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 5 6 from glycoproteins 867:879 arg1 release 825:831 efficient release 815:831 efficient release of the reduced O-glycans from the glycoproteins 815:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 3 7 theme glycosylation 423:435 arg1 analysis 403:410 Comprehensive analysis 389:410 Comprehensive analysis of protein glycosylation 389:435 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 7 8 theme third 1225:1229 arg1 technique 1231:1239 the third technique 1221:1239 the third technique 1221:1239 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 4 9 theme N-glycosidase 651:663 arg1 F 665:665 peptide N-glycosidase F 643:665 peptide N-glycosidase F 643:665 In contrast, N-glycans can be readily released non-reductively from peptide backbones using an enzyme such as peptide N-glycosidase F. 28091715 6 10 theme fusion 965:970 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 5 11 theme sensitive 690:698 arg1 protocol 700:707 a highly sensitive protocol 681:707 a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins 681:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 0 12 theme sensitive 7:15 arg1 labelling 31:39 Highly sensitive glycosylamine labelling 0:39 Highly sensitive glycosylamine labelling of O-glycans 0:52 Highly sensitive glycosylamine labelling of O-glycans using non-reductive β-elimination. 28091715 3 13 dep release 474:480 arg1 the 470:472 the 470:472 the 470:472 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 6 14 gly glycopeptide 1059:1070 arg2 glycopeptide 1059:1070 glycopeptide mapping 1059:1078 glycopeptide mapping 1059:1078 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 7 15 theme major 1142:1146 arg1 O-glycans 1148:1156 major O-glycans 1142:1156 major O-glycans 1142:1156 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 7 16 theme O-glycans 1148:1156 arg1 structures 1128:1137 The structures 1124:1137 The structures of major O-glycans and low-abundance O-glycans 1124:1184 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 6 17 theme glycopeptide 1059:1070 arg1 mapping 1072:1078 glycopeptide mapping 1059:1078 glycopeptide mapping 1059:1078 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 17 theme glycopeptide 1059:1070 arg1 methods 1051:1057 the analytical methods 1036:1057 the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling 1036:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 18 dep methods 1051:1057 arg1 methods 1051:1057 the analytical methods 1036:1057 the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling 1036:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 18 dep methods 1051:1057 arg1 mapping 1072:1078 glycopeptide mapping 1059:1078 glycopeptide mapping 1059:1078 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 18 dep methods 1051:1057 arg1 labelling 1086:1094 2-AB labelling 1081:1094 2-AB labelling 1081:1094 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 18 dep methods 1051:1057 arg1 labelling 1113:1121 RapiFluor-MS labelling 1100:1121 RapiFluor-MS labelling 1100:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 7 19 theme low-abundance 1162:1174 arg1 O-glycans 1176:1184 low-abundance O-glycans 1162:1184 low-abundance O-glycans 1162:1184 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 2 20 theme quality 347:353 arg1 profile 310:316 The glycan profile 299:316 The glycan profile 299:316 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 2 20 theme quality 347:353 arg1 attribute 355:363 a critical quality attribute 336:363 a critical quality attribute of a biopharmaceutical 336:386 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 3 21 theme O-glycans 522:530 arg1 separation 494:503 separation 494:503 separation 494:503 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 3 21 theme O-glycans 522:530 arg1 labelling 483:491 labelling 483:491 labelling 483:491 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 3 21 theme O-glycans 522:530 arg1 release 474:480 release 474:480 release 474:480 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 3 21 theme O-glycans 522:530 arg1 detection 509:517 detection 509:517 detection 509:517 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 0 22 theme glycosylamine 17:29 arg1 labelling 31:39 Highly sensitive glycosylamine labelling 0:39 Highly sensitive glycosylamine labelling of O-glycans 0:52 Highly sensitive glycosylamine labelling of O-glycans using non-reductive β-elimination. 28091715 6 23 theme analytical 1040:1049 arg1 labelling 1113:1121 RapiFluor-MS labelling 1100:1121 RapiFluor-MS labelling 1100:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 23 theme analytical 1040:1049 arg1 mapping 1072:1078 glycopeptide mapping 1059:1078 glycopeptide mapping 1059:1078 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 23 theme analytical 1040:1049 arg1 labelling 1086:1094 2-AB labelling 1081:1094 2-AB labelling 1081:1094 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 23 theme analytical 1040:1049 arg1 methods 1051:1057 the analytical methods 1036:1057 the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling 1036:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 7 24 theme O-glycans 1176:1184 arg1 structures 1128:1137 The structures 1124:1137 The structures of major O-glycans and low-abundance O-glycans 1124:1184 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 6 25 theme Ig 961:962 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 2 26 theme critical 338:345 arg1 profile 310:316 The glycan profile 299:316 The glycan profile 299:316 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 2 26 theme critical 338:345 arg1 attribute 355:363 a critical quality attribute 336:363 a critical quality attribute of a biopharmaceutical 336:386 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 6 27 used used 890:893 arg2 we 887:888 we 887:888 we 887:888 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 0 28 theme O-glycans 44:52 arg1 labelling 31:39 Highly sensitive glycosylamine labelling 0:39 Highly sensitive glycosylamine labelling of O-glycans 0:52 Highly sensitive glycosylamine labelling of O-glycans using non-reductive β-elimination. 28091715 5 29 theme non-enzyme 790:799 arg1 treatment 801:809 a non-enzyme treatment 788:809 a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins 788:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 1 30 theme between-protein 243:257 arg1 interactions 259:270 the between-protein interactions 239:270 the between-protein interactions that maintain homeostasis 239:296 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 6 31 theme G 958:958 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 32 theme O-glycan 1005:1012 arg1 analysis 1014:1021 O-glycan analysis 1005:1021 O-glycan analysis 1005:1021 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 0 33 theme non-reductive 60:72 arg1 β-elimination 74:86 non-reductive β-elimination 60:86 non-reductive β-elimination 60:86 Highly sensitive glycosylamine labelling of O-glycans using non-reductive β-elimination. 28091715 4 34 attach released 571:578 arg1 backbones 609:617 peptide backbones 601:617 peptide backbones using an enzyme such as peptide N-glycosidase F 601:665 In contrast, N-glycans can be readily released non-reductively from peptide backbones using an enzyme such as peptide N-glycosidase F. 28091715 4 34 attach released 571:578 arg2 N-glycans 546:554 N-glycans 546:554 N-glycans 546:554 In contrast, N-glycans can be readily released non-reductively from peptide backbones using an enzyme such as peptide N-glycosidase F. 28091715 5 35 theme O-glycan 756:763 arg1 analysis 765:772 O-glycan analysis 756:772 O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins 756:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 6 36 theme protein 933:939 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 37 dep lymphocyte 911:920 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 37 dep lymphocyte 911:920 arg1 fetuin 984:989 fetuin 984:989 fetuin 984:989 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 4 38 theme peptide 601:607 arg1 backbones 609:617 peptide backbones 601:617 peptide backbones using an enzyme such as peptide N-glycosidase F 601:665 In contrast, N-glycans can be readily released non-reductively from peptide backbones using an enzyme such as peptide N-glycosidase F. 28091715 6 39 theme associated 922:931 arg1 protein 972:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 associated protein 4-immunoglobulin G (Ig) fusion protein 922:978 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 5 40 gly glycoproteins 867:879 arg1 glycoproteins 867:879 the glycoproteins 863:879 the glycoproteins 863:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 5 41 theme reduced 840:846 arg1 O-glycans 848:856 the reduced O-glycans 836:856 the reduced O-glycans 836:856 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28091715 1 42 theme important 146:154 arg1 modifications 182:194 the most important posttranslational protein modifications 137:194 the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis 137:296 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 1 42 theme important 146:154 arg1 glycans 125:131 glycans 125:131 glycans 125:131 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 1 43 theme posttranslational 156:172 arg1 modifications 182:194 the most important posttranslational protein modifications 137:194 the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis 137:296 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 1 43 theme posttranslational 156:172 arg1 glycans 125:131 glycans 125:131 glycans 125:131 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 2 44 theme biopharmaceutical 370:386 arg1 profile 310:316 The glycan profile 299:316 The glycan profile 299:316 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 2 44 theme biopharmaceutical 370:386 arg1 attribute 355:363 a critical quality attribute 336:363 a critical quality attribute of a biopharmaceutical 336:386 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 6 45 theme T 909:909 arg1 models 994:999 models 994:999 models for O-glycan analysis 994:1021 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 45 theme T 909:909 arg1 lymphocyte 911:920 the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin 895:989 the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin 895:989 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 2 46 theme glycan 303:308 arg1 attribute 355:363 a critical quality attribute 336:363 a critical quality attribute of a biopharmaceutical 336:386 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 2 46 theme glycan 303:308 arg1 profile 310:316 The glycan profile 299:316 The glycan profile 299:316 The glycan profile may be defined as a critical quality attribute of a biopharmaceutical. 28091715 1 47 theme protein 174:180 arg1 modifications 182:194 the most important posttranslational protein modifications 137:194 the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis 137:296 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 1 47 theme protein 174:180 arg1 glycans 125:131 glycans 125:131 glycans 125:131 When developing biopharmaceuticals, glycans are the most important posttranslational protein modifications that must be addressed because they affect the between-protein interactions that maintain homeostasis. 28091715 6 48 theme RapiFluor-MS 1100:1111 arg1 methods 1051:1057 the analytical methods 1036:1057 the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling 1036:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 6 48 theme RapiFluor-MS 1100:1111 arg1 labelling 1113:1121 RapiFluor-MS labelling 1100:1121 RapiFluor-MS labelling 1100:1121 Here we used the cytotoxic T lymphocyte associated protein 4-immunoglobulin G (Ig) fusion protein and fetuin as models for O-glycan analysis and compared the analytical methods glycopeptide mapping, 2-AB labelling and RapiFluor-MS labelling. 28091715 3 49 theme Comprehensive 389:401 arg1 analysis 403:410 Comprehensive analysis 389:410 Comprehensive analysis of protein glycosylation 389:435 Comprehensive analysis of protein glycosylation must overcome challenges such as the release, labelling, separation and detection of O-glycans. 28091715 7 50 theme high 1276:1279 arg1 sensitivity 1281:1291 high sensitivity 1276:1291 high sensitivity 1276:1291 The structures of major O-glycans and low-abundance O-glycans were successfully identified using the third technique, which detected the O-glycans with high sensitivity. 28091715 5 51 theme efficient 815:823 arg1 release 825:831 efficient release 815:831 efficient release of the reduced O-glycans from the glycoproteins 815:879 We developed a highly sensitive protocol using RapiFluor-MS to label glycosylamines for O-glycan analysis combined with a non-enzyme treatment for efficient release of the reduced O-glycans from the glycoproteins. 28188786 6 0 theme conventional 978:989 arg1 induction 996:1004 the conventional IPTG induction 974:1004 the conventional IPTG induction 974:1004 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 4 1 theme 10th 620:623 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 1 theme 10th 620:623 arg1 FN3 660:662 FN3 660:662 FN3 660:662 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 1 theme 10th 620:623 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 1 theme 10th 620:623 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 1 2 link N-linked 282:289 arg1 glycosylation 299:311 N-linked protein glycosylation 282:311 N-linked protein glycosylation 282:311 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 2 link N-linked 282:289 arg1 pgl 277:279 pgl 277:279 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 3 3 theme N-glycosylated 531:544 arg1 proteins 546:553 N-glycosylated proteins 531:553 N-glycosylated proteins with high efficiency 531:574 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 4 4 theme fibronectin 631:641 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 4 theme fibronectin 631:641 arg1 FN3 660:662 FN3 660:662 FN3 660:662 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 4 theme fibronectin 631:641 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 4 theme fibronectin 631:641 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 1 5 from jejuni 339:344 arg1 locus 314:318 the pgl (N-linked protein glycosylation) locus 273:318 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 6 theme protein 291:297 arg1 glycosylation 299:311 N-linked protein glycosylation 282:311 N-linked protein glycosylation 282:311 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 6 theme protein 291:297 arg1 pgl 277:279 pgl 277:279 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 9 7 theme large-scale 1709:1719 arg1 production 1721:1730 the large-scale production 1705:1730 the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells 1705:1790 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 4 8 theme native 685:690 arg1 DFNRSK 713:718 native glycosylation sequon DFNRSK 685:718 native glycosylation sequon DFNRSK 685:718 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 6 9 theme glycosylation 935:947 arg1 efficiency 921:930 Increased efficiency 911:930 Increased efficiency of glycosylation 911:947 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 4 10 theme sequon 706:711 arg1 DFNRSK 713:718 native glycosylation sequon DFNRSK 685:718 native glycosylation sequon DFNRSK 685:718 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 1 11 gly N-glycosylated 234:247 arg1 protein 249:255 N-glycosylated protein 234:255 N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 234:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 12 theme successful 209:218 arg1 production 220:229 the successful production 205:229 the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 205:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 9 13 theme exogenous 1740:1748 arg1 proteins 1766:1773 pure exogenous N-glycosylation proteins 1735:1773 pure exogenous N-glycosylation proteins 1735:1773 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 3 14 theme auto-induction 593:606 arg1 method 608:613 an optimized auto-induction method 580:613 an optimized auto-induction method 580:613 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 5 15 theme anti-FLAG 888:896 arg1 antibody 901:908 anti-FLAG M1 antibody 888:908 anti-FLAG M1 antibody 888:908 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 9 16 from production 1721:1730 arg1 cells 1786:1790 E. coli cells 1778:1790 E. coli cells 1778:1790 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 9 17 theme proteins 1766:1773 arg1 production 1721:1730 the large-scale production 1705:1730 the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells 1705:1790 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 1 18 theme protein 249:255 arg1 production 220:229 the successful production 205:229 the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 205:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 6 19 theme auto-induction 1009:1022 arg1 method 1024:1029 auto-induction method 1009:1029 auto-induction method 1009:1029 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 9 20 theme E. coli 1778:1784 arg1 cells 1786:1790 E. coli cells 1778:1790 E. coli cells 1778:1790 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 8 21 theme lsg 1481:1483 arg1 locus 1485:1489 the reconstituted lsg locus 1463:1489 the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni 1463:1541 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 1 22 from production 220:229 arg1 E. coli 260:266 E. coli 260:266 E. coli 260:266 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 23 from E. coli 260:266 arg1 production 220:229 the successful production 205:229 the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 205:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 24 theme promising 145:153 arg1 cells 115:119 Escherichia coli cells 98:119 Escherichia coli cells 98:119 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 24 theme promising 145:153 arg1 hosts 155:159 promising hosts 145:159 promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 145:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 7 25 theme auto-induction 1347:1360 arg1 procedures 1362:1371 our optimized auto-induction procedures 1333:1371 our optimized auto-induction procedures 1333:1371 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 0 26 from efficiency 24:33 arg1 coli 74:77 Escherichia coli 62:77 Escherichia coli 62:77 Increased glycosylation efficiency of recombinant proteins in Escherichia coli by auto-induction. 28188786 4 27 theme DQNAT 734:738 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 27 theme DQNAT 734:738 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 27 theme DQNAT 734:738 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 7 28 theme local 1239:1243 arg1 conformation 1256:1267 local structural conformation 1239:1267 local structural conformation 1239:1267 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 1 29 theme N-glycosylated 175:188 arg1 proteins 190:197 N-glycosylated proteins 175:197 N-glycosylated proteins 175:197 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 6 30 from % 1094:1094 arg1 efficiencies 1066:1077 the glycosylation efficiencies 1048:1077 the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively 1048:1126 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 8 31 used utilized 1548:1555 arg2 locus 1485:1489 the reconstituted lsg locus 1463:1489 the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni 1463:1541 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 8 32 from influenzae 1508:1517 arg1 locus 1485:1489 the reconstituted lsg locus 1463:1489 the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni 1463:1541 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 6 33 theme glycosylation 1052:1064 arg1 efficiencies 1066:1077 the glycosylation efficiencies 1048:1077 the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively 1048:1126 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 3 34 gly N-glycosylated 531:544 arg1 proteins 546:553 N-glycosylated proteins 531:553 N-glycosylated proteins with high efficiency 531:574 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 0 35 theme proteins 50:57 arg1 efficiency 24:33 glycosylation efficiency 10:33 glycosylation efficiency of recombinant proteins in Escherichia coli 10:77 Increased glycosylation efficiency of recombinant proteins in Escherichia coli by auto-induction. 28188786 7 36 theme optimized 1337:1345 arg1 procedures 1362:1371 our optimized auto-induction procedures 1333:1371 our optimized auto-induction procedures 1333:1371 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 7 37 theme FN3 1209:1211 arg1 loop 1201:1204 the loop 1197:1204 the loop of FN3 (the acceptor sequon with local structural conformation) 1197:1268 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 4 38 theme acceptor 785:792 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 38 theme acceptor 785:792 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 38 theme acceptor 785:792 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 8 39 from PglB 1523:1526 arg1 locus 1485:1489 the reconstituted lsg locus 1463:1489 the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni 1463:1541 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 1 40 with protein 249:255 arg1 locus 314:318 the pgl (N-linked protein glycosylation) locus 273:318 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 7 41 theme 80 1326:1327 arg1 % 1321:1321 % 1321:1321 % 1321:1321 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 8 42 theme 100 1574:1576 arg1 % 1577:1577 % 1577:1577 % 1577:1577 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 2 43 theme inefficient 436:446 arg1 glycosylation 455:467 inefficient glycan glycosylation 436:467 inefficient glycan glycosylation 436:467 However, one hurdle in producing N-glycosylated proteins in large scale using E. coli is inefficient glycan glycosylation. 28188786 0 44 theme recombinant 38:48 arg1 proteins 50:57 recombinant proteins 38:57 recombinant proteins 38:57 Increased glycosylation efficiency of recombinant proteins in Escherichia coli by auto-induction. 28188786 1 45 theme Campylobacter 325:337 arg1 jejuni 339:344 Campylobacter jejuni 325:344 Campylobacter jejuni 325:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 5 46 theme glycosylation 824:836 arg1 efficiencies 838:849 The resulting glycosylation efficiencies 810:849 The resulting glycosylation efficiencies 810:849 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 6 47 theme Increased 911:919 arg1 efficiency 921:930 Increased efficiency 911:930 Increased efficiency of glycosylation 911:947 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 8 48 theme glycosylation 1579:1591 arg1 efficiency 1593:1602 100% glycosylation efficiency 1574:1602 100% glycosylation efficiency 1574:1602 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 7 49 gly glycosylation 1173:1185 arg2 sequon 1187:1192 the glycosylation sequon 1169:1192 the glycosylation sequon 1169:1192 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 7 50 theme glycosylation 1173:1185 arg1 sequon 1187:1192 the glycosylation sequon 1169:1192 the glycosylation sequon 1169:1192 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 8 51 theme auto-induction 1440:1453 arg1 method 1455:1460 the optimized auto-induction method 1426:1460 the optimized auto-induction method 1426:1460 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 7 52 dep % 1321:1321 arg1 to 1323:1324 to 1323:1324 to 1323:1324 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 4 53 theme type 643:646 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 53 theme type 643:646 arg1 FN3 660:662 FN3 660:662 FN3 660:662 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 53 theme type 643:646 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 53 theme type 643:646 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 3 54 theme proteins 546:553 arg1 production 517:526 the production 513:526 the production of N-glycosylated proteins with high efficiency 513:574 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 4 55 from sequon 740:745 arg1 C-terminus 750:759 C-terminus 750:759 C-terminus 750:759 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 6 56 theme IPTG 991:994 arg1 induction 996:1004 the conventional IPTG induction 974:1004 the conventional IPTG induction 974:1004 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 9 57 theme auto-induction 1667:1680 arg1 method 1682:1687 the optimized auto-induction method 1653:1687 the optimized auto-induction method 1653:1687 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 7 58 theme glycosylation 1275:1287 arg1 efficiency 1289:1298 the glycosylation efficiency 1271:1298 the glycosylation efficiency 1271:1298 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 2 59 theme N-glycosylated 380:393 arg1 proteins 395:402 N-glycosylated proteins 380:402 N-glycosylated proteins 380:402 However, one hurdle in producing N-glycosylated proteins in large scale using E. coli is inefficient glycan glycosylation. 28188786 1 60 theme pgl 277:279 arg1 locus 314:318 the pgl (N-linked protein glycosylation) locus 273:318 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 4 61 with sequon 740:745 arg1 linker 775:780 flexible linker 766:780 flexible linker 766:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 3 62 with proteins 546:553 arg1 efficiency 565:574 high efficiency 560:574 high efficiency 560:574 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 1 63 theme N-linked 282:289 arg1 glycosylation 299:311 N-linked protein glycosylation 282:311 N-linked protein glycosylation 282:311 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 63 theme N-linked 282:289 arg1 pgl 277:279 pgl 277:279 the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 273:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 64 dep Escherichia 98:108 arg1 coli 110:113 coli 110:113 coli 110:113 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 4 65 theme human 625:629 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 65 theme human 625:629 arg1 FN3 660:662 FN3 660:662 FN3 660:662 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 65 theme human 625:629 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 65 theme human 625:629 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 3 66 theme high 560:563 arg1 efficiency 565:574 high efficiency 560:574 high efficiency 560:574 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 9 67 theme pure 1735:1738 arg1 proteins 1766:1773 pure exogenous N-glycosylation proteins 1735:1773 pure exogenous N-glycosylation proteins 1735:1773 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 5 68 with blots 877:881 arg1 antibody 901:908 anti-FLAG M1 antibody 888:908 anti-FLAG M1 antibody 888:908 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 4 69 theme glycosylation 692:704 arg1 DFNRSK 713:718 native glycosylation sequon DFNRSK 685:718 native glycosylation sequon DFNRSK 685:718 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 3 70 theme optimized 583:591 arg1 method 608:613 an optimized auto-induction method 580:613 an optimized auto-induction method 580:613 In this study, we developed a strategy for the production of N-glycosylated proteins with high efficiency via an optimized auto-induction method. 28188786 9 71 theme N-glycosylation 1750:1764 arg1 proteins 1766:1773 pure exogenous N-glycosylation proteins 1735:1773 pure exogenous N-glycosylation proteins 1735:1773 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 8 72 theme reconstituted 1467:1479 arg1 locus 1485:1489 the reconstituted lsg locus 1463:1489 the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni 1463:1541 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 5 73 theme M1 898:899 arg1 antibody 901:908 anti-FLAG M1 antibody 888:908 anti-FLAG M1 antibody 888:908 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 1 74 from protein 249:255 arg1 E. coli 260:266 E. coli 260:266 E. coli 260:266 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 7 75 with sequon 1227:1232 arg1 conformation 1256:1267 local structural conformation 1239:1267 local structural conformation 1239:1267 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 1 76 gly N-glycosylated 175:188 arg1 proteins 190:197 N-glycosylated proteins 175:197 N-glycosylated proteins 175:197 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 7 77 theme structural 1245:1254 arg1 conformation 1256:1267 local structural conformation 1239:1267 local structural conformation 1239:1267 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 6 78 from % 1086:1086 arg1 efficiencies 1066:1077 the glycosylation efficiencies 1048:1077 the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively 1048:1126 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 7 79 theme acceptor 1218:1225 arg1 sequon 1227:1232 the acceptor sequon 1214:1232 the acceptor sequon with local structural conformation 1214:1267 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 7 80 dep FN3 1209:1211 arg1 sequon 1227:1232 the acceptor sequon 1214:1232 the acceptor sequon with local structural conformation 1214:1267 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 4 81 theme protein 794:800 arg1 models 802:807 acceptor protein models 785:807 acceptor protein models 785:807 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 81 theme protein 794:800 arg1 domain 652:657 The 10th human fibronectin type III domain 616:657 The 10th human fibronectin type III domain (FN3) 616:663 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 4 81 theme protein 794:800 arg1 sequon 740:745 DQNAT sequon 734:745 DQNAT sequon in C-terminus with flexible linker 734:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 6 82 dep % 1094:1094 arg1 % 1104:1104 up to 90% 1096:1104 75% up to 90% 1092:1104 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 6 83 dep 90 1102:1103 arg1 to 1099:1100 to 1099:1100 to 1099:1100 Increased efficiency of glycosylation was obtained by changing the conventional IPTG induction to auto-induction method, which increased the glycosylation efficiencies from 60% and 75% up to 90% and 100% respectively. 28188786 0 84 theme glycosylation 10:22 arg1 efficiency 24:33 glycosylation efficiency 10:33 glycosylation efficiency of recombinant proteins in Escherichia coli 10:77 Increased glycosylation efficiency of recombinant proteins in Escherichia coli by auto-induction. 28188786 1 85 theme Escherichia 98:108 arg1 cells 115:119 Escherichia coli cells 98:119 Escherichia coli cells 98:119 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 1 85 theme Escherichia 98:108 arg1 hosts 155:159 promising hosts 145:159 promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 145:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 4 86 theme flexible 766:773 arg1 linker 775:780 flexible linker 766:780 flexible linker 766:780 The 10th human fibronectin type III domain (FN3) was engineered with native glycosylation sequon DFNRSK and optimized DQNAT sequon in C-terminus with flexible linker as acceptor protein models. 28188786 2 87 theme large 407:411 arg1 scale 413:417 large scale 407:417 large scale using E. coli 407:431 However, one hurdle in producing N-glycosylated proteins in large scale using E. coli is inefficient glycan glycosylation. 28188786 5 88 theme Western 869:875 arg1 blots 877:881 Western blots 869:881 Western blots with anti-FLAG M1 antibody 869:908 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 8 89 theme % 1577:1577 arg1 efficiency 1593:1602 100% glycosylation efficiency 1574:1602 100% glycosylation efficiency 1574:1602 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 2 90 theme glycan 448:453 arg1 glycosylation 455:467 inefficient glycan glycosylation 436:467 inefficient glycan glycosylation 436:467 However, one hurdle in producing N-glycosylated proteins in large scale using E. coli is inefficient glycan glycosylation. 28188786 5 91 theme resulting 814:822 arg1 efficiencies 838:849 The resulting glycosylation efficiencies 810:849 The resulting glycosylation efficiencies 810:849 The resulting glycosylation efficiencies were confirmed by Western blots with anti-FLAG M1 antibody. 28188786 1 92 theme N-glycosylated 234:247 arg1 protein 249:255 N-glycosylated protein 234:255 N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni 234:344 Escherichia coli cells have been considered as promising hosts for producing N-glycosylated proteins since the successful production of N-glycosylated protein in E. coli with the pgl (N-linked protein glycosylation) locus from Campylobacter jejuni. 28188786 7 93 theme 35 1319:1320 arg1 % 1321:1321 % 1321:1321 % 1321:1321 Moreover, in the condition of inserting the glycosylation sequon in the loop of FN3 (the acceptor sequon with local structural conformation), the glycosylation efficiency was increased from 35% to 80% by our optimized auto-induction procedures. 28188786 2 94 gly N-glycosylated 380:393 arg1 proteins 395:402 N-glycosylated proteins 380:402 N-glycosylated proteins 380:402 However, one hurdle in producing N-glycosylated proteins in large scale using E. coli is inefficient glycan glycosylation. 28188786 8 95 theme general 1403:1409 arg1 application 1411:1421 general application 1403:1421 general application of the optimized auto-induction method 1403:1460 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 9 96 theme quantitative 1626:1637 arg1 evidence 1639:1646 quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells 1626:1790 quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells 1626:1790 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 8 97 theme optimized 1430:1438 arg1 method 1455:1460 the optimized auto-induction method 1426:1460 the optimized auto-induction method 1426:1460 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28188786 9 98 theme optimized 1657:1665 arg1 method 1682:1687 the optimized auto-induction method 1653:1687 the optimized auto-induction method 1653:1687 Our studies provided quantitative evidence that the optimized auto-induction method will facilitate the large-scale production of pure exogenous N-glycosylation proteins in E. coli cells. 28188786 8 99 theme method 1455:1460 arg1 application 1411:1421 general application 1403:1421 general application of the optimized auto-induction method 1403:1460 To justify the potential for general application of the optimized auto-induction method, the reconstituted lsg locus from Haemophilus influenzae and PglB from C. jejuni were utilized, and this led to 100% glycosylation efficiency. 28096352 1 0 theme modifications 234:246 arg1 addition 150:157 the addition 146:157 the addition of covalently linked sugars (glycosylation) 146:201 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 0 theme modifications 234:246 arg1 modifications 234:246 the most abundant modifications 216:246 the most abundant modifications 216:246 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 0 theme modifications 234:246 arg1 one 209:211 one 209:211 one 209:211 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 2 1 theme essential 336:344 arg1 hydrolysis 270:279 the hydrolysis 266:279 the hydrolysis of protein peptide bonds by peptidases 266:318 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 1 theme essential 336:344 arg1 process 328:334 a process 326:334 a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation 326:477 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 3 2 theme carbohydrate-specific 730:750 arg1 subsites 752:759 carbohydrate-specific subsites 730:759 carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery 730:819 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 0 3 link protein-linked 15:28 arg1 glycans 30:36 protein-linked glycans 15:36 protein-linked glycans 15:36 Recognition of protein-linked glycans as a determinant of peptidase activity. 28096352 3 4 theme crystallographic 543:558 arg1 structures 560:569 X-ray crystallographic structures 537:569 X-ray crystallographic structures 537:569 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 1 5 theme proteins 99:106 arg1 majority 87:94 The vast majority 78:94 The vast majority of proteins 78:106 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 3 6 theme peptidase 791:799 arg1 machinery 811:819 their peptidase catalytic machinery 785:819 their peptidase catalytic machinery 785:819 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 4 7 theme glycan 938:943 arg1 modifications 945:957 particular glycan modifications 927:957 particular glycan modifications 927:957 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 3 8 theme adjacent 773:780 arg1 subsites 752:759 carbohydrate-specific subsites 730:759 carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery 730:819 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 4 9 theme particular 927:936 arg1 modifications 945:957 particular glycan modifications 927:957 particular glycan modifications 927:957 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 3 10 theme X-ray 537:541 arg1 structures 560:569 X-ray crystallographic structures 537:569 X-ray crystallographic structures 537:569 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 1 11 theme linked 173:178 arg1 sugars 180:185 covalently linked sugars 162:185 covalently linked sugars (glycosylation) 162:201 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 11 theme linked 173:178 arg1 glycosylation 188:200 glycosylation 188:200 glycosylation 188:200 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 12 theme vast 82:85 arg1 majority 87:94 The vast majority 78:94 The vast majority of proteins 78:106 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 13 theme sugars 180:185 arg1 addition 150:157 the addition 146:157 the addition of covalently linked sugars (glycosylation) 146:201 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 13 theme sugars 180:185 arg1 modifications 234:246 the most abundant modifications 216:246 the most abundant modifications 216:246 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 13 theme sugars 180:185 arg1 one 209:211 one 209:211 one 209:211 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 2 14 theme posttranslational 422:438 arg1 modifications 440:452 posttranslational modifications 422:452 posttranslational modifications 422:452 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 14 theme posttranslational 422:438 arg1 glycosylation 465:477 glycosylation 465:477 glycosylation 465:477 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 1 15 link linked 173:178 arg1 sugars 180:185 covalently linked sugars 162:185 covalently linked sugars (glycosylation) 162:201 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 1 15 link linked 173:178 arg1 glycosylation 188:200 glycosylation 188:200 glycosylation 188:200 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 28096352 2 16 from life 353:356 arg1 details 384:390 the fundamental details 368:390 the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation 368:477 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 16 from life 353:356 arg1 Earth 361:365 Earth 361:365 Earth 361:365 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 17 theme bonds 300:304 arg1 hydrolysis 270:279 the hydrolysis 266:279 the hydrolysis of protein peptide bonds by peptidases 266:318 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 17 theme bonds 300:304 arg1 process 328:334 a process 326:334 a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation 326:477 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 0 18 theme glycans 30:36 arg1 Recognition 0:10 Recognition 0:10 Recognition of protein-linked glycans as a determinant of peptidase activity 0:75 Recognition of protein-linked glycans as a determinant of peptidase activity. 28096352 4 19 theme glycosylated 1022:1033 arg1 residue 1035:1041 the glycosylated residue 1018:1041 the glycosylated residue 1018:1041 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 2 20 theme peptide 292:298 arg1 bonds 300:304 protein peptide bonds 284:304 protein peptide bonds 284:304 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 0 21 theme protein-linked 15:28 arg1 glycans 30:36 protein-linked glycans 15:36 protein-linked glycans 15:36 Recognition of protein-linked glycans as a determinant of peptidase activity. 28096352 3 22 theme catalytic 801:809 arg1 machinery 811:819 their peptidase catalytic machinery 785:819 their peptidase catalytic machinery 785:819 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 3 23 theme specific 672:679 arg1 recognition 681:691 the specific recognition 668:691 the specific recognition of O-linked glycan modifications 668:724 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 2 24 theme protein 284:290 arg1 bonds 300:304 protein peptide bonds 284:304 protein peptide bonds 284:304 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 4 25 theme protein-specific 895:910 arg1 adaptations 912:922 protein-specific adaptations 895:922 protein-specific adaptations 895:922 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 2 26 theme fundamental 372:382 arg1 details 384:390 the fundamental details 368:390 the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation 368:477 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 2 26 theme fundamental 372:382 arg1 Earth 361:365 Earth 361:365 Earth 361:365 However, despite the hydrolysis of protein peptide bonds by peptidases being a process essential to all life on Earth, the fundamental details of how peptidases accommodate posttranslational modifications, including glycosylation, has not been addressed. 28096352 5 27 theme molecular 1136:1144 arg1 understanding 1146:1158 a molecular understanding 1134:1158 a molecular understanding of glycoprotein degradation 1134:1186 This insight builds upon the paradigm of how peptidases recognize substrates and provides a molecular understanding of glycoprotein degradation. 28096352 4 28 theme peptide 983:989 arg1 bond 991:994 the peptide bond 979:994 the peptide bond immediately preceding the glycosylated residue 979:1041 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 3 29 theme glycan 705:710 arg1 modifications 712:724 O-linked glycan modifications 696:724 O-linked glycan modifications 696:724 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 5 30 theme glycoprotein 1163:1174 arg1 degradation 1176:1186 glycoprotein degradation 1163:1186 glycoprotein degradation 1163:1186 This insight builds upon the paradigm of how peptidases recognize substrates and provides a molecular understanding of glycoprotein degradation. 28096352 4 31 gly glycosylated 1022:1033 arg1 residue 1035:1041 the glycosylated residue 1018:1041 the glycosylated residue 1018:1041 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 3 32 theme modifications 712:724 arg1 recognition 681:691 the specific recognition 668:691 the specific recognition of O-linked glycan modifications 668:724 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 3 33 theme related 634:640 arg1 metallopeptidases 642:658 three structurally related metallopeptidases 615:658 three structurally related metallopeptidases 615:658 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 3 34 link O-linked 696:703 arg1 modifications 712:724 O-linked glycan modifications 696:724 O-linked glycan modifications 696:724 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 5 35 theme degradation 1176:1186 arg1 understanding 1146:1158 a molecular understanding 1134:1158 a molecular understanding of glycoprotein degradation 1134:1186 This insight builds upon the paradigm of how peptidases recognize substrates and provides a molecular understanding of glycoprotein degradation. 28096352 4 36 theme different 866:874 arg1 glycans 876:882 different glycans 866:882 different glycans 866:882 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 0 37 theme activity 68:75 arg1 determinant 43:53 a determinant 41:53 a determinant of peptidase activity 41:75 Recognition of protein-linked glycans as a determinant of peptidase activity. 28096352 3 38 theme O-linked 696:703 arg1 modifications 712:724 O-linked glycan modifications 696:724 O-linked glycan modifications 696:724 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 3 39 theme biochemical 512:522 arg1 analyses 524:531 biochemical analyses 512:531 biochemical analyses 512:531 Through biochemical analyses and X-ray crystallographic structures we show that to hydrolyze their substrates, three structurally related metallopeptidases require the specific recognition of O-linked glycan modifications via carbohydrate-specific subsites immediately adjacent to their peptidase catalytic machinery. 28096352 0 40 theme peptidase 58:66 arg1 activity 68:75 peptidase activity 58:75 peptidase activity 58:75 Recognition of protein-linked glycans as a determinant of peptidase activity. 28096352 4 41 dep showed 843:848 arg1 cleaved 971:977 cleaved 971:977 cleaved the peptide bond immediately preceding the glycosylated residue 971:1041 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 4 41 dep showed 843:848 arg1 revealing 885:893 revealing 885:893 revealing protein-specific adaptations to particular glycan modifications 885:957 The three peptidases showed selectivity for different glycans, revealing protein-specific adaptations to particular glycan modifications, yet always cleaved the peptide bond immediately preceding the glycosylated residue. 28096352 5 42 gly glycoprotein 1163:1174 arg1 glycoprotein 1163:1174 glycoprotein degradation 1163:1186 glycoprotein degradation 1163:1186 This insight builds upon the paradigm of how peptidases recognize substrates and provides a molecular understanding of glycoprotein degradation. 28096352 1 43 theme abundant 225:232 arg1 modifications 234:246 the most abundant modifications 216:246 the most abundant modifications 216:246 The vast majority of proteins are posttranslationally altered, with the addition of covalently linked sugars (glycosylation) being one of the most abundant modifications. 27798070 3 0 from milieu 473:478 arg1 modifications 399:411 dynamic modifications 391:411 dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu 391:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 9 1 theme cell 1692:1695 arg1 glycans 1705:1711 cell surface glycans 1692:1711 cell surface glycans 1692:1711 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 5 2 theme sequential 920:929 arg1 spectrometry 943:954 sequential tandem mass spectrometry 920:954 sequential tandem mass spectrometry 920:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 3 theme activities 675:684 arg1 evaluation 605:614 an evaluation 602:614 an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures 602:854 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 4 theme dynamic 391:397 arg1 modifications 399:411 dynamic modifications 391:411 dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu 391:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 9 5 theme surface 1697:1703 arg1 glycans 1705:1711 cell surface glycans 1692:1711 cell surface glycans 1692:1711 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 5 6 theme tandem 931:936 arg1 spectrometry 943:954 sequential tandem mass spectrometry 920:954 sequential tandem mass spectrometry 920:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 7 theme fucosyltransferase 656:673 arg1 activities 675:684 fucosyltransferase activities 656:684 fucosyltransferase activities 656:684 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 8 theme distal 427:432 arg1 surfaces 439:446 distal cell surfaces 427:446 distal cell surfaces 427:446 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 8 9 theme mouse 1533:1537 arg1 platelets 1539:1547 mouse platelets 1533:1547 mouse platelets 1533:1547 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 8 10 theme sialyltransferases 1471:1488 arg1 amounts 1416:1422 large amounts 1410:1422 large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets 1410:1547 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 8 10 theme sialyltransferases 1471:1488 arg1 sialyltransferases 1471:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 4 11 theme GlcNAc 750:755 arg1 motifs 742:747 the four common terminal glycan precursor motifs 700:747 the four common terminal glycan precursor motifs 700:747 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 11 theme GlcNAc 750:755 arg1 monomer 757:763 GlcNAc monomer 750:763 GlcNAc monomer 750:763 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 12 theme cell 434:437 arg1 surfaces 439:446 distal cell surfaces 427:446 distal cell surfaces 427:446 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 3 13 from surfaces 439:446 arg1 modifications 399:411 dynamic modifications 391:411 dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu 391:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 4 14 theme Gal 799:801 arg1 GalNAc 806:811 Gal(β4)GlcNAc and Gal(β3)GalNAc 781:811 GalNAc 806:811 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 14 theme Gal 799:801 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 5 15 from analysis 908:915 arg1 Data 857:860 Data 857:860 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry 857:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 2 16 theme inflammatory 340:351 arg1 conditions 353:362 inflammatory conditions 340:362 inflammatory conditions 340:362 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 4 17 theme Gal 781:783 arg1 GlcNAc 788:793 Gal(β4)GlcNAc and Gal(β3)GalNAc 781:811 GlcNAc 788:793 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 17 theme Gal 781:783 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 18 theme glycans 416:422 arg1 modifications 399:411 dynamic modifications 391:411 dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu 391:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 8 19 dep Gal 1457:1459 arg1 to 1454:1455 to 1454:1455 to 1454:1455 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 8 19 dep Gal 1457:1459 arg1 β4 1461:1462 β4 1461:1462 β4 1461:1462 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 5 20 theme detailed 891:898 arg1 analysis 908:915 detailed product analysis 891:915 detailed product analysis by sequential tandem mass spectrometry 891:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 3 21 theme present 513:519 arg1 activities 502:511 the glycan-modifying activities 481:511 the glycan-modifying activities present in systemic circulation 481:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 9 22 theme cell 1723:1726 arg1 behavior 1728:1735 cell behavior 1723:1735 cell behavior 1723:1735 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 4 23 theme β3 803:804 arg1 GalNAc 806:811 Gal(β4)GlcNAc and Gal(β3)GalNAc 781:811 GalNAc 806:811 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 23 theme β3 803:804 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 9 24 theme circulatory 1590:1600 arg1 glycosyltransferases 1602:1621 circulatory glycosyltransferases 1590:1621 circulatory glycosyltransferases 1590:1621 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 0 25 theme blood 12:16 arg1 glycosyltransferases 39:58 blood and platelets supply glycosyltransferases 12:58 blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation 12:108 Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. 27798070 8 26 theme Gal 1457:1459 arg1 GlcNAc 1464:1469 the α3-sialyl to Gal(β4)GlcNAc 1440:1469 the α3-sialyl to Gal(β4)GlcNAc 1440:1469 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 3 27 attach present 513:519 arg1 circulation 533:543 systemic circulation 524:543 systemic circulation 524:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 3 27 attach present 513:519 arg2 activities 502:511 the glycan-modifying activities 481:511 the glycan-modifying activities present in systemic circulation 481:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 9 28 theme glycosyltransferases 1602:1621 arg1 capability 1576:1585 the capability 1572:1585 the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior 1572:1735 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 3 29 theme glycan-modifying 485:500 arg1 activities 502:511 the glycan-modifying activities 481:511 the glycan-modifying activities present in systemic circulation 481:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 4 30 theme β4 785:786 arg1 GlcNAc 788:793 Gal(β4)GlcNAc and Gal(β3)GalNAc 781:811 GlcNAc 788:793 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 30 theme β4 785:786 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 5 31 theme product 900:906 arg1 analysis 908:915 detailed product analysis 891:915 detailed product analysis by sequential tandem mass spectrometry 891:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 0 32 theme platelets 22:30 arg1 glycosyltransferases 39:58 blood and platelets supply glycosyltransferases 12:58 blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation 12:108 Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. 27798070 4 33 theme sialyl- 631:637 arg1 evaluation 605:614 an evaluation 602:614 an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures 602:854 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 34 theme glycan 838:843 arg1 structures 845:854 more complex glycan structures 825:854 more complex glycan structures 825:854 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 8 35 theme ST6Gal-1 1427:1434 arg1 sialyltransferases 1471:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 5 36 theme glycan 1043:1048 arg1 motifs 1050:1055 the well-recognized and important glycan motifs 1009:1055 the well-recognized and important glycan motifs 1009:1055 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 36 theme glycan 1043:1048 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 37 theme glycan 725:730 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 37 theme glycan 725:730 arg1 motifs 742:747 the four common terminal glycan precursor motifs 700:747 the four common terminal glycan precursor motifs 700:747 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 37 theme glycan 725:730 arg1 monomer 757:763 GlcNAc monomer 750:763 GlcNAc monomer 750:763 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 8 38 theme large 1410:1414 arg1 amounts 1416:1422 large amounts 1410:1422 large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets 1410:1547 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 8 38 theme large 1410:1414 arg1 sialyltransferases 1471:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 3 39 from glycans 416:422 arg1 milieu 473:478 the extracellular milieu 455:478 the extracellular milieu 455:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 3 39 from glycans 416:422 arg1 surfaces 439:446 distal cell surfaces 427:446 distal cell surfaces 427:446 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 8 40 theme Human 1372:1376 arg1 carriers 1397:1404 carriers 1397:1404 carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets 1397:1547 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 8 40 theme Human 1372:1376 arg1 platelets 1378:1386 Human platelets 1372:1386 Human platelets 1372:1386 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 5 41 contain has 972:974 arg1 blood 966:970 blood 966:970 blood 966:970 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 41 contain has 972:974 arg2 capacity 980:987 the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 976:1116 the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 976:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 42 theme terminal 716:723 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 42 theme terminal 716:723 arg1 motifs 742:747 the four common terminal glycan precursor motifs 700:747 the four common terminal glycan precursor motifs 700:747 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 42 theme terminal 716:723 arg1 monomer 757:763 GlcNAc monomer 750:763 GlcNAc monomer 750:763 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 43 theme galactosyl- 640:650 arg1 evaluation 605:614 an evaluation 602:614 an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures 602:854 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 5 44 theme H- 1093:1094 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 45 theme complex 830:836 arg1 structures 845:854 more complex glycan structures 825:854 more complex glycan structures 825:854 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 6 46 theme mouse 1208:1212 arg1 β4GalT 1282:1287 β4GalT 1282:1287 β4GalT 1282:1287 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 46 theme mouse 1208:1212 arg1 platelets 1214:1222 human and mouse platelets 1198:1222 human and mouse platelets 1198:1222 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 46 theme mouse 1208:1212 arg1 carriers 1238:1245 important carriers 1228:1245 important carriers 1228:1245 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 46 theme mouse 1208:1212 arg1 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 3 47 theme systemic 524:531 arg1 circulation 533:543 systemic circulation 524:543 systemic circulation 524:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 0 48 theme supply 32:37 arg1 glycosyltransferases 39:58 blood and platelets supply glycosyltransferases 12:58 blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation 12:108 Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. 27798070 6 49 theme human 1198:1202 arg1 β4GalT 1282:1287 β4GalT 1282:1287 β4GalT 1282:1287 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 49 theme human 1198:1202 arg1 platelets 1214:1222 human and mouse platelets 1198:1222 human and mouse platelets 1198:1222 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 49 theme human 1198:1202 arg1 carriers 1238:1245 important carriers 1228:1245 important carriers 1228:1245 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 49 theme human 1198:1202 arg1 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 5 50 theme motifs 1050:1055 arg1 motifs 1050:1055 the well-recognized and important glycan motifs 1009:1055 the well-recognized and important glycan motifs 1009:1055 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 50 theme motifs 1050:1055 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 50 theme motifs 1050:1055 arg1 many 1001:1004 many 1001:1004 many 1001:1004 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 51 theme radioisotope 867:878 arg1 assays 880:885 radioisotope assays 867:885 radioisotope assays 867:885 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 52 theme precursor 732:740 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 52 theme precursor 732:740 arg1 motifs 742:747 the four common terminal glycan precursor motifs 700:747 the four common terminal glycan precursor motifs 700:747 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 52 theme precursor 732:740 arg1 monomer 757:763 GlcNAc monomer 750:763 GlcNAc monomer 750:763 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 53 theme extracellular 459:471 arg1 milieu 473:478 the extracellular milieu 455:478 the extracellular milieu 455:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 5 54 theme Sialyl-T 1100:1107 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 3 55 from circulation 533:543 arg1 present 513:519 present 513:519 present 513:519 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 5 56 theme mass 938:941 arg1 spectrometry 943:954 sequential tandem mass spectrometry 920:954 sequential tandem mass spectrometry 920:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 57 theme blood-borne 619:629 arg1 sialyl- 631:637 blood-borne sialyl- 619:637 blood-borne sialyl- 619:637 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 2 58 theme blood-borne 239:249 arg1 glycosyltransferases 251:270 these blood-borne glycosyltransferases 233:270 these blood-borne glycosyltransferases 233:270 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 2 59 theme glycosyltransferases 251:270 arg1 glycosyltransferases 251:270 these blood-borne glycosyltransferases 233:270 these blood-borne glycosyltransferases 233:270 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 2 59 theme glycosyltransferases 251:270 arg1 Many 225:228 Many 225:228 Many 225:228 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 6 60 theme glycosyltransferases 1139:1158 arg1 many 1125:1128 many 1125:1128 many 1125:1128 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 60 theme glycosyltransferases 1139:1158 arg1 glycosyltransferases 1139:1158 these glycosyltransferases 1133:1158 these glycosyltransferases 1133:1158 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 8 61 theme GlcNAc 1464:1469 arg1 sialyltransferases 1471:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases 1427:1488 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 5 62 theme important 1033:1041 arg1 motifs 1050:1055 the well-recognized and important glycan motifs 1009:1055 the well-recognized and important glycan motifs 1009:1055 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 62 theme important 1033:1041 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 63 theme Lewis 1072:1076 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 6 64 theme important 1228:1236 arg1 β4GalT 1282:1287 β4GalT 1282:1287 β4GalT 1282:1287 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 64 theme important 1228:1236 arg1 platelets 1214:1222 human and mouse platelets 1198:1222 human and mouse platelets 1198:1222 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 64 theme important 1228:1236 arg1 carriers 1238:1245 important carriers 1228:1245 important carriers 1228:1245 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 6 64 theme important 1228:1236 arg1 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 ST3Gal-1 1269:1276 While many of these glycosyltransferases are freely circulating in the plasma, human and mouse platelets are important carriers for others, including ST3Gal-1 and β4GalT. 27798070 1 65 theme intracellular 161:173 arg1 apparatus 185:193 the intracellular secretory apparatus 157:193 the intracellular secretory apparatus 157:193 Glycosyltransferases, usually residing within the intracellular secretory apparatus, also circulate in the blood. 27798070 5 66 from assays 880:885 arg1 Data 857:860 Data 857:860 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry 857:954 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 67 dep Gal 766:768 arg1 GlcNAc 773:778 GlcNAc 773:778 GlcNAc 773:778 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 3 68 from modifications 399:411 arg1 milieu 473:478 the extracellular milieu 455:478 the extracellular milieu 455:478 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 3 68 from modifications 399:411 arg1 surfaces 439:446 distal cell surfaces 427:446 distal cell surfaces 427:446 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 5 69 theme well-recognized 1013:1027 arg1 motifs 1050:1055 the well-recognized and important glycan motifs 1009:1055 the well-recognized and important glycan motifs 1009:1055 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 5 69 theme well-recognized 1013:1027 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 1 70 theme secretory 175:183 arg1 apparatus 185:193 the intracellular secretory apparatus 157:193 the intracellular secretory apparatus 157:193 Glycosyltransferases, usually residing within the intracellular secretory apparatus, also circulate in the blood. 27798070 0 71 theme extracellular 82:94 arg1 glycosylation 96:108 extrinsic extracellular glycosylation 72:108 extrinsic extracellular glycosylation 72:108 Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. 27798070 5 72 theme sialyl-Lewis 1079:1090 arg1 antigens 1109:1116 the Lewis, sialyl-Lewis, H- and Sialyl-T antigens 1068:1116 antigens 1109:1116 Data from radioisotope assays and detailed product analysis by sequential tandem mass spectrometry show that blood has the capacity to generate many of the well-recognized and important glycan motifs, including the Lewis, sialyl-Lewis, H- and Sialyl-T antigens. 27798070 4 73 theme common 709:714 arg1 Gal 766:768 Gal 766:768 Gal 766:768 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 73 theme common 709:714 arg1 motifs 742:747 the four common terminal glycan precursor motifs 700:747 the four common terminal glycan precursor motifs 700:747 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 4 73 theme common 709:714 arg1 monomer 757:763 GlcNAc monomer 750:763 GlcNAc monomer 750:763 Here, we describe an evaluation of blood-borne sialyl-, galactosyl- and fucosyltransferase activities that act upon the four common terminal glycan precursor motifs, GlcNAc monomer, Gal(β3)GlcNAc, Gal(β4)GlcNAc and Gal(β3)GalNAc, to produce more complex glycan structures. 27798070 0 74 theme extrinsic 72:80 arg1 glycosylation 96:108 extrinsic extracellular glycosylation 72:108 extrinsic extracellular glycosylation 72:108 Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. 27798070 2 75 theme pathological 292:303 arg1 states 305:310 pathological states 292:310 pathological states 292:310 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 2 75 theme pathological 292:303 arg1 malignancies 323:334 malignancies 323:334 malignancies 323:334 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 2 75 theme pathological 292:303 arg1 conditions 353:362 inflammatory conditions 340:362 inflammatory conditions 340:362 Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. 27798070 3 76 from present 513:519 arg1 circulation 533:543 systemic circulation 524:543 systemic circulation 524:543 Despite the potential for dynamic modifications of glycans on distal cell surfaces and in the extracellular milieu, the glycan-modifying activities present in systemic circulation have not been systematically examined. 27798070 8 77 from platelets 1539:1547 arg1 absent 1523:1528 absent 1523:1528 absent 1523:1528 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 27798070 9 78 theme platelet 1660:1667 arg1 activation 1669:1678 platelet activation 1660:1678 platelet activation 1660:1678 This study highlights the capability of circulatory glycosyltransferases, which are dynamically controlled by platelet activation, to remodel cell surface glycans and alter cell behavior. 27798070 8 79 from absent 1523:1528 arg1 platelets 1539:1547 mouse platelets 1533:1547 mouse platelets 1533:1547 Human platelets are also carriers for large amounts of ST6Gal-1 and the α3-sialyl to Gal(β4)GlcNAc sialyltransferases, both of which are conspicuously absent in mouse platelets. 28494147 4 0 theme impaired 1030:1037 arg1 interactions 1039:1050 the impaired interactions 1026:1050 the impaired interactions 1026:1050 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 3 1 theme influence 725:733 arg1 understanding 704:716 a better understanding 695:716 a better understanding of the influence of O-glycan structure on the molecule's physical stability 695:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 3 2 from stability 784:792 arg1 understanding 704:716 a better understanding 695:716 a better understanding of the influence of O-glycan structure on the molecule's physical stability 695:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 1 3 theme effects 159:165 arg1 effects 159:165 site-specific and glycan-specific effects 125:165 site-specific and glycan-specific effects 125:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 1 3 theme effects 159:165 arg1 variety 114:120 a variety 112:120 a variety of site-specific and glycan-specific effects 112:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 4 4 theme new 908:910 arg1 interactions 920:931 new bonding interactions 908:931 new bonding interactions 908:931 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 4 5 theme bonding 912:918 arg1 interactions 920:931 new bonding interactions 908:931 new bonding interactions 908:931 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 3 6 theme structural 541:550 arg1 features 552:559 the structural features 537:559 the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 537:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 4 7 theme decreased 984:992 arg1 stability 994:1002 the decreased stability 980:1002 the decreased stability 980:1002 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 3 8 theme structure 747:755 arg1 influence 725:733 the influence 721:733 the influence of O-glycan structure on the molecule's physical stability 721:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 1 9 theme Protein 67:73 arg1 glycosylation 75:87 Protein glycosylation 67:87 Protein glycosylation 67:87 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 2 10 theme structural 268:277 arg1 changes 279:285 the structural changes 264:285 the structural changes that occur 264:296 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 5 11 theme general 1153:1159 arg1 mechanism 1161:1169 a powerful and potentially general mechanism 1126:1169 a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering 1126:1234 This type of knowledge provides a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering. 28494147 2 12 gly glycopeptide 368:379 arg2 glycopeptide 368:379 the glycopeptide 364:379 the glycopeptide 364:379 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 13 gly glycoproteins 494:506 arg1 glycoproteins 494:506 glycoproteins 494:506 glycoproteins 494:506 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 14 theme glycoproteins 494:506 arg1 stability 481:489 the stability 477:489 the stability of glycoproteins 477:506 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 4 15 theme O-glycans 868:876 arg1 effects 857:863 the previously observed stabilizing effects 821:863 the previously observed stabilizing effects of O-glycans 821:876 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 2 16 theme glycopeptide 368:379 arg1 steps 394:398 valuable steps 385:398 valuable steps 385:398 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 16 theme glycopeptide 368:379 arg1 properties 350:359 the physical properties 337:359 the physical properties of the glycopeptide 337:379 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 3 17 link reesei-derived 628:641 arg1 cellobiohydrolase 652:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 3 18 theme O-glycan 738:745 arg1 structure 747:755 O-glycan structure 738:755 O-glycan structure 738:755 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 3 19 theme O-glycosylated 568:581 arg1 module 604:609 the O-glycosylated carbohydrate-binding module 564:609 the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 564:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 0 20 theme Structural 0:9 arg1 Insight 11:17 Structural Insight 0:17 Structural Insight into the Stabilizing Effect of O-Glycosylation 0:64 Structural Insight into the Stabilizing Effect of O-Glycosylation. 28494147 3 21 theme carbohydrate-binding 583:602 arg1 module 604:609 the O-glycosylated carbohydrate-binding module 564:609 the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 564:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 3 22 theme physical 775:782 arg1 stability 784:792 the molecule's physical stability 760:792 the molecule's physical stability 760:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 2 23 dep those 326:330 arg1 steps 394:398 valuable steps 385:398 valuable steps 385:398 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 23 dep those 326:330 arg1 properties 350:359 the physical properties 337:359 the physical properties of the glycopeptide 337:379 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 1 24 theme molecular 184:192 arg1 elusive 241:247 elusive 241:247 elusive 241:247 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 1 24 theme molecular 184:192 arg1 logic 194:198 the molecular logic 180:198 the molecular logic that leads to such observations 180:230 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 3 25 from understanding 704:716 arg1 stability 784:792 the molecule's physical stability 760:792 the molecule's physical stability 760:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 4 26 theme stabilizing 845:855 arg1 effects 857:863 the previously observed stabilizing effects 821:863 the previously observed stabilizing effects of O-glycans 821:876 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 0 27 theme Stabilizing 28:38 arg1 Effect 40:45 the Stabilizing Effect 24:45 the Stabilizing Effect of O-Glycosylation 24:64 Structural Insight into the Stabilizing Effect of O-Glycosylation. 28494147 1 28 contain have 107:110 arg1 glycosylation 75:87 Protein glycosylation 67:87 Protein glycosylation 67:87 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 1 28 contain have 107:110 arg2 effects 159:165 site-specific and glycan-specific effects 125:165 site-specific and glycan-specific effects 125:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 1 28 contain have 107:110 arg2 variety 114:120 a variety 112:120 a variety of site-specific and glycan-specific effects 112:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 5 29 theme powerful 1128:1135 arg1 mechanism 1161:1169 a powerful and potentially general mechanism 1126:1169 a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering 1126:1234 This type of knowledge provides a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering. 28494147 3 30 theme Trichoderma 616:626 arg1 cellobiohydrolase 652:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 2 31 theme valuable 385:392 arg1 steps 394:398 valuable steps 385:398 valuable steps 385:398 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 31 theme valuable 385:392 arg1 properties 350:359 the physical properties 337:359 the physical properties of the glycopeptide 337:379 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 0 32 theme O-Glycosylation 50:64 arg1 Effect 40:45 the Stabilizing Effect 24:45 the Stabilizing Effect of O-Glycosylation 24:64 Structural Insight into the Stabilizing Effect of O-Glycosylation. 28494147 5 33 theme proteins 1202:1209 arg1 stability 1189:1197 the stability 1185:1197 the stability of proteins 1185:1209 This type of knowledge provides a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering. 28494147 3 34 theme module 604:609 arg1 features 552:559 the structural features 537:559 the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 537:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 2 35 theme specific 433:440 arg1 patterns 456:463 specific glycosylation patterns 433:463 specific glycosylation patterns 433:463 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 4 36 theme interactions 920:931 arg1 introduction 892:903 the introduction 888:903 the introduction of new bonding interactions to the structure 888:948 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 1 37 theme glycan-specific 143:157 arg1 effects 159:165 site-specific and glycan-specific effects 125:165 site-specific and glycan-specific effects 125:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 3 38 gly O-glycosylated 568:581 arg1 module 604:609 the O-glycosylated carbohydrate-binding module 564:609 the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 564:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 3 39 theme better 697:702 arg1 understanding 704:716 a better understanding 695:716 a better understanding of the influence of O-glycan structure on the molecule's physical stability 695:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 4 40 theme observed 836:843 arg1 effects 857:863 the previously observed stabilizing effects 821:863 the previously observed stabilizing effects of O-glycans 821:876 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 3 41 from influence 725:733 arg1 stability 784:792 the molecule's physical stability 760:792 the molecule's physical stability 760:792 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 4 42 theme conformational 1066:1079 arg1 flexibility 1081:1091 conformational flexibility 1066:1091 conformational flexibility 1066:1091 Our results indicate that the previously observed stabilizing effects of O-glycans come from the introduction of new bonding interactions to the structure and increased rigidity, while the decreased stability seemed to result from the impaired interactions and increased conformational flexibility. 28494147 3 43 theme cellobiohydrolase 652:668 arg1 module 604:609 the O-glycosylated carbohydrate-binding module 564:609 the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase 564:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 1 44 theme site-specific 125:137 arg1 effects 159:165 site-specific and glycan-specific effects 125:165 site-specific and glycan-specific effects 125:165 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 5 45 theme knowledge 1107:1115 arg1 type 1099:1102 This type 1094:1102 This type of knowledge 1094:1115 This type of knowledge provides a powerful and potentially general mechanism for improving the stability of proteins through glycoengineering. 28494147 3 46 theme reesei-derived 628:641 arg1 cellobiohydrolase 652:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 1 47 theme such 214:217 arg1 observations 219:230 such observations 214:230 such observations 214:230 Protein glycosylation has been shown to have a variety of site-specific and glycan-specific effects, but so far, the molecular logic that leads to such observations has been elusive. 28494147 2 48 theme glycosylation 442:454 arg1 patterns 456:463 specific glycosylation patterns 433:463 specific glycosylation patterns 433:463 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 3 49 theme Family 643:648 arg1 cellobiohydrolase 652:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 a Trichoderma reesei-derived Family 7 cellobiohydrolase 614:668 By systematically comparing the structural features of the O-glycosylated carbohydrate-binding module of a Trichoderma reesei-derived Family 7 cellobiohydrolase, we were able to develop a better understanding of the influence of O-glycan structure on the molecule's physical stability. 28494147 2 50 theme physical 341:348 arg1 steps 394:398 valuable steps 385:398 valuable steps 385:398 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 28494147 2 50 theme physical 341:348 arg1 properties 350:359 the physical properties 337:359 the physical properties of the glycopeptide 337:379 Understanding the structural changes that occur and being able to correlate those with the physical properties of the glycopeptide are valuable steps toward being able to predict how specific glycosylation patterns will affect the stability of glycoproteins. 24303873 8 0 used used 1198:1201 arg2 clusterings 1139:1149 Hierarchical clusterings 1126:1149 Hierarchical clusterings based on Pearson correlation coefficients 1126:1191 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 12 1 theme potential 1873:1881 arg1 applications 1883:1894 potential applications 1873:1894 potential applications toward cancer diagnosis and direction of treatment 1873:1945 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 2 2 theme biopsy 342:347 arg1 results 349:355 traditional biopsy results 330:355 traditional biopsy results 330:355 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 1 3 theme histochemical 234:246 arg1 staining 271:278 histochemical or immunohistochemical staining 234:278 histochemical or immunohistochemical staining 234:278 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 4 4 theme isomer-sensitive 593:608 arg1 nano-LC/MS 610:619 isomer-sensitive nano-LC/MS 593:619 isomer-sensitive nano-LC/MS 593:619 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 1 5 theme immunohistochemical 251:269 arg1 staining 271:278 histochemical or immunohistochemical staining 234:278 histochemical or immunohistochemical staining 234:278 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 8 6 theme separate 1226:1233 arg1 line 1245:1248 separate each cell line 1226:1248 separate each cell line according to originating organ 1226:1279 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 12 7 theme MS-based 1851:1858 arg1 biopsy 1860:1865 an MS-based biopsy 1848:1865 an MS-based biopsy 1848:1865 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 1 8 used used 144:147 arg2 biopsies 121:128 biopsies 121:128 biopsies 121:128 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 3 9 theme alternate 537:545 arg1 route 547:551 an alternate route 534:551 an alternate route toward cancer diagnosis 534:575 Glycomic profiling of the cell membrane offers an alternate route toward cancer diagnosis. 24303873 8 10 theme cell 1240:1243 arg1 line 1245:1248 separate each cell line 1226:1248 separate each cell line according to originating organ 1226:1279 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 6 11 theme carbon 901:906 arg1 nano-LC/MS 908:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 4 12 theme N-glycan 684:691 arg1 structures 693:702 the different N-glycan structures 670:702 the different N-glycan structures present on cancer cell membranes 670:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 2 13 theme significant 301:311 arg1 number 313:318 a significant number 299:318 a significant number of cases 299:327 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 7 14 theme most 1107:1110 arg1 compositions 1112:1123 most compositions 1107:1123 most compositions 1107:1123 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 3 15 theme cell 513:516 arg1 membrane 518:525 the cell membrane 509:525 the cell membrane 509:525 Glycomic profiling of the cell membrane offers an alternate route toward cancer diagnosis. 24303873 8 16 theme originating 1263:1273 arg1 organ 1275:1279 originating organ 1263:1279 originating organ 1263:1279 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 6 17 theme native 968:973 arg1 N-glycans 975:983 the native N-glycans 964:983 the native N-glycans 964:983 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 5 18 theme Membrane 738:745 arg1 N-glycans 747:755 Membrane N-glycans 738:755 Membrane N-glycans 738:755 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 7 19 theme peaks 1055:1059 arg1 isomers 1095:1101 multiple isomers 1086:1101 multiple isomers for most compositions 1086:1123 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 7 19 theme peaks 1055:1059 arg1 hundreds 1036:1043 hundreds 1036:1043 hundreds 1036:1043 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 5 20 theme cancer 863:868 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 12 21 theme cell 1793:1796 arg1 glycosylation 1807:1819 cancer cell membrane glycosylation 1786:1819 cancer cell membrane glycosylation 1786:1819 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 9 22 theme cell 1454:1457 arg1 lines 1459:1463 most cell lines 1449:1463 most cell lines 1449:1463 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 4 23 theme present 704:710 arg1 structures 693:702 the different N-glycan structures 670:702 the different N-glycan structures present on cancer cell membranes 670:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 6 24 dep identify 941:948 arg1 separate 931:938 separate 931:938 separate 931:938 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 5 25 theme ovarian 840:846 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 7 26 theme N-glycan 1006:1013 arg1 profiling 1015:1023 Structure-sensitive N-glycan profiling 986:1023 Structure-sensitive N-glycan profiling 986:1023 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 8 27 theme Hierarchical 1126:1137 arg1 clusterings 1139:1149 Hierarchical clusterings 1126:1149 Hierarchical clusterings based on Pearson correlation coefficients 1126:1191 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 4 28 theme cancer 715:720 arg1 membranes 727:735 cancer cell membranes 715:735 cancer cell membranes 715:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 0 29 theme membrane 70:77 arg1 profiling 88:96 plasma membrane N-glycan profiling 63:96 plasma membrane N-glycan profiling 63:96 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 9 30 theme broad 1345:1349 arg1 classes 1358:1364 broad glycan classes 1345:1364 broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.) 1345:1446 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 2 31 from number 313:318 arg1 inconclusive 368:379 inconclusive 368:379 inconclusive 368:379 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 9 32 dep mannose 1378:1384 arg1 e.g. 1367:1370 e.g. 1367:1370 e.g. 1367:1370 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 9 33 theme classes 1358:1364 arg1 abundances 1331:1340 the relative abundances 1318:1340 the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.) 1318:1446 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 9 34 dep sialylated 1430:1439 arg1 complex/hybrid 1415:1428 complex/hybrid 1415:1428 complex/hybrid 1415:1428 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 10 35 theme related 1507:1513 arg1 lines 1520:1524 More closely related cell lines 1494:1524 More closely related cell lines 1494:1524 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 5 36 theme cervical 830:837 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 9 37 theme high 1373:1376 arg1 mannose 1378:1384 high mannose 1373:1384 high mannose 1373:1384 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 0 38 theme cancer 19:24 arg1 origin 31:36 cancer cell origin 19:36 cancer cell origin 19:36 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 6 39 theme graphitized 889:899 arg1 nano-LC/MS 908:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 1 40 theme clinical 102:109 arg1 settings 111:118 clinical settings 102:118 clinical settings 102:118 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 2 41 theme full 399:402 arg1 differentiation 412:426 full subtype differentiation 399:426 full subtype differentiation 399:426 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 0 42 theme origin 31:36 arg1 Differentiation 0:14 Differentiation 0:14 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 0:97 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 5 43 theme breast 816:821 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 6 44 theme Chip-based 871:880 arg1 nano-LC/MS 908:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 12 45 theme cancer 1903:1908 arg1 diagnosis 1910:1918 cancer diagnosis 1903:1918 cancer diagnosis 1903:1918 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 11 46 theme primary 1664:1670 arg1 origins 1679:1685 primary cancer origins 1664:1685 primary cancer origins 1664:1685 Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. 24303873 2 47 theme traditional 330:340 arg1 results 349:355 traditional biopsy results 330:355 traditional biopsy results 330:355 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 5 48 theme lung 824:827 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 2 49 theme cases 323:327 arg1 number 313:318 a significant number 299:318 a significant number of cases 299:327 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 8 50 theme Pearson 1160:1166 arg1 coefficients 1180:1191 Pearson correlation coefficients 1160:1191 Pearson correlation coefficients 1160:1191 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 10 51 theme glycans 1612:1618 arg1 abundances 1587:1596 the abundances 1583:1596 the abundances of individual glycans 1583:1618 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 11 52 theme molecular 1691:1699 arg1 subtypes 1701:1708 molecular subtypes 1691:1708 molecular subtypes 1691:1708 Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. 24303873 3 53 theme membrane 518:525 arg1 profiling 496:504 Glycomic profiling 487:504 Glycomic profiling of the cell membrane 487:525 Glycomic profiling of the cell membrane offers an alternate route toward cancer diagnosis. 24303873 5 54 theme various 796:802 arg1 subtypes 804:811 various subtypes 796:811 various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer 796:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 9 55 theme most 1449:1452 arg1 lines 1459:1463 most cell lines 1449:1463 most cell lines 1449:1463 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 4 56 attach present 704:710 arg2 structures 693:702 the different N-glycan structures 670:702 the different N-glycan structures present on cancer cell membranes 670:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 4 56 attach present 704:710 arg1 membranes 727:735 cancer cell membranes 715:735 cancer cell membranes 715:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 1 57 theme cancer 162:167 arg1 type 169:172 cancer type 162:172 cancer type 162:172 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 0 58 theme molecular 42:50 arg1 subtype 52:58 molecular subtype 42:58 molecular subtype 42:58 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 8 59 theme disease 1285:1291 arg1 subtype 1293:1299 disease subtype 1285:1299 disease subtype 1285:1299 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 6 60 used used 923:926 arg2 nano-LC/MS 908:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 11 61 theme characteristic 1630:1643 arg1 profiles 1654:1661 characteristic N-glycan profiles 1630:1661 characteristic N-glycan profiles 1630:1661 Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. 24303873 8 62 theme each 1235:1238 arg1 line 1245:1248 separate each cell line 1226:1248 separate each cell line according to originating organ 1226:1279 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 4 63 theme different 674:682 arg1 structures 693:702 the different N-glycan structures 670:702 the different N-glycan structures present on cancer cell membranes 670:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 4 64 used used 625:628 arg2 nano-LC/MS 610:619 isomer-sensitive nano-LC/MS 593:619 isomer-sensitive nano-LC/MS 593:619 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 3 65 theme cancer 560:565 arg1 diagnosis 567:575 cancer diagnosis 560:575 cancer diagnosis 560:575 Glycomic profiling of the cell membrane offers an alternate route toward cancer diagnosis. 24303873 7 66 theme multiple 1086:1093 arg1 isomers 1095:1101 multiple isomers 1086:1101 multiple isomers for most compositions 1086:1123 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 5 67 theme lymphatic 853:861 arg1 cancer 863:868 lymphatic cancer 853:868 lymphatic cancer 853:868 Membrane N-glycans were extracted from cells representing various subtypes of breast, lung, cervical, ovarian, and lymphatic cancer. 24303873 4 68 theme detailed 649:656 arg1 profiles 658:665 detailed profiles 649:665 detailed profiles of the different N-glycan structures present on cancer cell membranes 649:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 4 69 theme cell 722:725 arg1 membranes 727:735 cancer cell membranes 715:735 cancer cell membranes 715:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 7 70 theme cell 1065:1068 arg1 line 1070:1073 cell line 1065:1073 cell line 1065:1073 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 7 71 theme glycan 1048:1053 arg1 peaks 1055:1059 glycan peaks 1048:1059 glycan peaks per cell line 1048:1073 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 12 72 theme cancer 1786:1791 arg1 glycosylation 1807:1819 cancer cell membrane glycosylation 1786:1819 cancer cell membrane glycosylation 1786:1819 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 12 73 from differences 1771:1781 arg1 glycosylation 1807:1819 cancer cell membrane glycosylation 1786:1819 cancer cell membrane glycosylation 1786:1819 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 0 74 theme plasma 63:68 arg1 profiling 88:96 plasma membrane N-glycan profiling 63:96 plasma membrane N-glycan profiling 63:96 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 9 75 theme relative 1322:1329 arg1 abundances 1331:1340 the relative abundances 1318:1340 the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.) 1318:1446 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 12 76 theme membrane 1798:1805 arg1 glycosylation 1807:1819 cancer cell membrane glycosylation 1786:1819 cancer cell membrane glycosylation 1786:1819 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 4 77 theme structures 693:702 arg1 profiles 658:665 detailed profiles 649:665 detailed profiles of the different N-glycan structures present on cancer cell membranes 649:735 In this study, isomer-sensitive nano-LC/MS was used to directly obtain detailed profiles of the different N-glycan structures present on cancer cell membranes. 24303873 0 78 theme N-glycan 79:86 arg1 profiling 88:96 plasma membrane N-glycan profiling 63:96 plasma membrane N-glycan profiling 63:96 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 12 79 theme treatment 1937:1945 arg1 direction 1924:1932 direction 1924:1932 direction 1924:1932 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 12 79 theme treatment 1937:1945 arg1 diagnosis 1910:1918 cancer diagnosis 1903:1918 cancer diagnosis 1903:1918 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 10 80 from differences 1568:1578 arg1 abundances 1587:1596 the abundances 1583:1596 the abundances of individual glycans 1583:1618 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 9 81 theme glycan 1351:1356 arg1 classes 1358:1364 broad glycan classes 1345:1364 broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.) 1345:1446 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 7 82 theme Structure-sensitive 986:1004 arg1 profiling 1015:1023 Structure-sensitive N-glycan profiling 986:1023 Structure-sensitive N-glycan profiling 986:1023 Structure-sensitive N-glycan profiling identified hundreds of glycan peaks per cell line, including multiple isomers for most compositions. 24303873 10 83 theme cell 1515:1518 arg1 lines 1520:1524 More closely related cell lines 1494:1524 More closely related cell lines 1494:1524 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 1 84 theme tumor 193:197 arg1 morphology 204:213 tumor cell morphology 193:213 tumor cell morphology 193:213 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 0 85 theme cell 26:29 arg1 origin 31:36 cancer cell origin 19:36 cancer cell origin 19:36 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 2 86 theme subtype 404:410 arg1 differentiation 412:426 full subtype differentiation 399:426 full subtype differentiation 399:426 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 6 87 theme porous 882:887 arg1 nano-LC/MS 908:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS 871:917 Chip-based porous graphitized carbon nano-LC/MS was used to separate, identify, and quantify the native N-glycans. 24303873 12 88 theme stark 1765:1769 arg1 differences 1771:1781 stark differences 1765:1781 stark differences in cancer cell membrane glycosylation 1765:1819 These results demonstrate that stark differences in cancer cell membrane glycosylation can be exploited to create an MS-based biopsy, with potential applications toward cancer diagnosis and direction of treatment. 24303873 2 89 theme ineffective 464:474 arg1 treatment 476:484 inefficient or ineffective treatment 449:484 inefficient or ineffective treatment 449:484 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 0 90 theme subtype 52:58 arg1 Differentiation 0:14 Differentiation 0:14 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 0:97 Differentiation of cancer cell origin and molecular subtype by plasma membrane N-glycan profiling. 24303873 2 91 from inconclusive 368:379 arg1 number 313:318 a significant number 299:318 a significant number of cases 299:327 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 2 92 theme inefficient 449:459 arg1 treatment 476:484 inefficient or ineffective treatment 449:484 inefficient or ineffective treatment 449:484 Unfortunately, in a significant number of cases, traditional biopsy results are either inconclusive or do not provide full subtype differentiation, possibly leading to inefficient or ineffective treatment. 24303873 9 93 dep classes 1358:1364 arg1 etc. 1442:1445 etc. 1442:1445 etc. 1442:1445 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 9 93 dep classes 1358:1364 arg1 fucosylated 1402:1412 fucosylated 1402:1412 fucosylated 1402:1412 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 9 93 dep classes 1358:1364 arg1 sialylated 1430:1439 sialylated 1430:1439 sialylated 1430:1439 Based simply on the relative abundances of broad glycan classes (e.g., high mannose, complex/hybrid fucosylated, complex/hybrid sialylated, etc.), most cell lines were readily differentiated. 24303873 11 94 theme N-glycan 1645:1652 arg1 profiles 1654:1661 characteristic N-glycan profiles 1630:1661 characteristic N-glycan profiles 1630:1661 Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. 24303873 8 95 theme correlation 1168:1178 arg1 coefficients 1180:1191 Pearson correlation coefficients 1160:1191 Pearson correlation coefficients 1160:1191 Hierarchical clusterings based on Pearson correlation coefficients were used to quickly compare and separate each cell line according to originating organ and disease subtype. 24303873 10 96 theme several-fold 1555:1566 arg1 differences 1568:1578 several-fold differences 1555:1578 several-fold differences in the abundances of individual glycans 1555:1618 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 3 97 theme Glycomic 487:494 arg1 profiling 496:504 Glycomic profiling 487:504 Glycomic profiling of the cell membrane 487:525 Glycomic profiling of the cell membrane offers an alternate route toward cancer diagnosis. 24303873 1 98 theme cell 199:202 arg1 morphology 204:213 tumor cell morphology 193:213 tumor cell morphology 193:213 In clinical settings, biopsies are routinely used to determine cancer type and grade based on tumor cell morphology, as determined via histochemical or immunohistochemical staining. 24303873 10 99 theme individual 1601:1610 arg1 glycans 1612:1618 individual glycans 1601:1618 individual glycans 1601:1618 More closely related cell lines were differentiated based on several-fold differences in the abundances of individual glycans. 24303873 11 100 theme cancer 1672:1677 arg1 origins 1679:1685 primary cancer origins 1664:1685 primary cancer origins 1664:1685 Based on characteristic N-glycan profiles, primary cancer origins and molecular subtypes could be distinguished. 26203194 5 0 from molecules 964:972 arg1 surface 1012:1018 the cell surface 1003:1018 the cell surface 1003:1018 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 1 from flexible 761:768 arg1 solution 773:780 solution 773:780 solution 773:780 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 2 from solution 773:780 arg1 terminus 730:737 The C terminus 724:737 The C terminus 724:737 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 2 from solution 773:780 arg1 flexible 761:768 flexible 761:768 flexible 761:768 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 1 3 theme cellular 173:180 arg1 pathways 192:199 several important cellular signaling pathways 155:199 several important cellular signaling pathways 155:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 0 4 from Aspects 11:17 arg1 Glypican-1 74:83 Human Glypican-1 68:83 Human Glypican-1 68:83 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 1 5 theme signaling 182:190 arg1 pathways 192:199 several important cellular signaling pathways 155:199 several important cellular signaling pathways 155:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 3 6 theme cell 396:399 arg1 membrane 401:408 the cell membrane 392:408 the cell membrane 392:408 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 3 7 attach attach 365:370 arg2 domain 353:358 the C-terminal domain 338:358 the C-terminal domain that attach the core protein to the cell membrane 338:408 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 3 7 attach attach 365:370 arg1 membrane 401:408 the cell membrane 392:408 the cell membrane 392:408 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 3 8 theme core 376:379 arg1 protein 381:387 the core protein 372:387 the core protein 372:387 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 0 9 from Region 58:63 arg1 Glypican-1 74:83 Human Glypican-1 68:83 Human Glypican-1 68:83 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 5 10 theme C 728:728 arg1 terminus 730:737 The C terminus 724:737 The C terminus 724:737 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 10 theme C 728:728 arg1 flexible 761:768 flexible 761:768 flexible 761:768 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 2 11 theme human 296:300 arg1 brain 302:306 adult human brain 290:306 adult human brain 290:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 3 12 theme Gpc1 434:437 arg1 structure 447:455 the Gpc1 crystal structure 430:455 the Gpc1 crystal structure 430:455 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 5 13 from substitution 1069:1080 arg1 apparatus 1095:1103 the Golgi apparatus 1085:1103 the Golgi apparatus 1085:1103 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 2 14 theme adult 290:294 arg1 brain 302:306 adult human brain 290:306 adult human brain 290:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 2 15 from proteoglycan 255:266 arg1 brain 302:306 adult human brain 290:306 adult human brain 290:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 2 15 from proteoglycan 255:266 arg1 developing 275:284 developing 275:284 developing 275:284 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 3 16 theme crystal 439:445 arg1 structure 447:455 the Gpc1 crystal structure 430:455 the Gpc1 crystal structure 430:455 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 5 17 theme heparan 1053:1059 arg1 sulfate 1061:1067 heparan sulfate 1053:1067 heparan sulfate substitution in the Golgi apparatus 1053:1103 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 3 18 theme C-terminal 342:351 arg1 domain 353:358 the C-terminal domain 338:358 the C-terminal domain that attach the core protein to the cell membrane 338:408 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 5 19 theme Gpc1 891:894 arg1 orthologs 896:904 Gpc1 orthologs 891:904 Gpc1 orthologs 891:904 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 0 20 theme Structural 0:9 arg1 Aspects 11:17 Structural Aspects 0:17 Structural Aspects of N-Glycosylations 0:37 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 4 21 theme chromatographic 547:561 arg1 approaches 563:572 chromatographic approaches 547:572 chromatographic approaches 547:572 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 2 22 dep developing 275:284 arg1 the 271:273 the 271:273 the 271:273 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 0 23 theme N-Glycosylations 22:37 arg1 Aspects 11:17 Structural Aspects 0:17 Structural Aspects of N-Glycosylations 0:37 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 0 23 theme N-Glycosylations 22:37 arg1 Region 58:63 the C-terminal Region 43:63 the C-terminal Region in Human Glypican-1 43:83 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 6 24 theme protein 1157:1163 arg1 stability 1165:1173 the protein stability 1153:1173 the protein stability 1153:1173 Furthermore, the N-glycans are shown to extend the protein stability and lifetime by protection against proteolysis and aggregation. 26203194 1 25 theme multifunctional 100:114 arg1 proteoglycans 129:141 multifunctional cell surface proteoglycans 100:141 multifunctional cell surface proteoglycans involved in several important cellular signaling pathways 100:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 1 25 theme multifunctional 100:114 arg1 Glypicans 86:94 Glypicans 86:94 Glypicans 86:94 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 1 26 theme cell 116:119 arg1 proteoglycans 129:141 multifunctional cell surface proteoglycans 100:141 multifunctional cell surface proteoglycans involved in several important cellular signaling pathways 100:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 1 26 theme cell 116:119 arg1 Glypicans 86:94 Glypicans 86:94 Glypicans 86:94 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 5 27 theme membrane 981:988 arg1 receptors 990:998 membrane receptors 981:998 membrane receptors 981:998 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 4 28 theme terminus 655:662 arg1 composition 591:601 composition 591:601 composition 591:601 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 28 theme terminus 655:662 arg1 structure 604:612 structure 604:612 structure 604:612 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 28 theme terminus 655:662 arg1 function 619:626 function 619:626 function 619:626 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 29 theme C 653:653 arg1 terminus 655:662 the C terminus 649:662 the C terminus 649:662 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 30 theme x-ray 525:529 arg1 scattering 531:540 small angle x-ray scattering 513:540 small angle x-ray scattering 513:540 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 2 31 theme sulfate 247:253 arg1 Glypican-1 202:211 Glypican-1 202:211 Glypican-1 (Gpc1) 202:218 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 2 31 theme sulfate 247:253 arg1 proteoglycan 255:266 the predominant heparan sulfate proteoglycan 223:266 the predominant heparan sulfate proteoglycan in the developing and adult human brain 223:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 4 32 theme small 513:517 arg1 scattering 531:540 small angle x-ray scattering 513:540 small angle x-ray scattering 513:540 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 33 theme N-glycans 635:643 arg1 composition 591:601 composition 591:601 composition 591:601 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 33 theme N-glycans 635:643 arg1 structure 604:612 structure 604:612 structure 604:612 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 33 theme N-glycans 635:643 arg1 function 619:626 function 619:626 function 619:626 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 4 34 theme angle 519:523 arg1 scattering 531:540 small angle x-ray scattering 513:540 small angle x-ray scattering 513:540 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 3 35 theme N-linked 317:324 arg1 glycans 326:332 The two N-linked glycans 309:332 The two N-linked glycans 309:332 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 2 36 theme heparan 239:245 arg1 Glypican-1 202:211 Glypican-1 202:211 Glypican-1 (Gpc1) 202:218 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 2 36 theme heparan 239:245 arg1 proteoglycan 255:266 the predominant heparan sulfate proteoglycan 223:266 the predominant heparan sulfate proteoglycan in the developing and adult human brain 223:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 1 37 theme surface 121:127 arg1 proteoglycans 129:141 multifunctional cell surface proteoglycans 100:141 multifunctional cell surface proteoglycans involved in several important cellular signaling pathways 100:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 1 37 theme surface 121:127 arg1 Glypicans 86:94 Glypicans 86:94 Glypicans 86:94 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 0 38 theme C-terminal 47:56 arg1 Region 58:63 the C-terminal Region 43:63 the C-terminal Region in Human Glypican-1 43:83 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 2 39 theme predominant 227:237 arg1 Glypican-1 202:211 Glypican-1 202:211 Glypican-1 (Gpc1) 202:218 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 2 39 theme predominant 227:237 arg1 proteoglycan 255:266 the predominant heparan sulfate proteoglycan 223:266 the predominant heparan sulfate proteoglycan in the developing and adult human brain 223:306 Glypican-1 (Gpc1) is the predominant heparan sulfate proteoglycan in the developing and adult human brain. 26203194 5 40 theme Golgi 1089:1093 arg1 apparatus 1095:1103 the Golgi apparatus 1085:1103 the Golgi apparatus 1085:1103 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 4 41 theme Gpc1 689:692 arg1 topology 677:684 the topology 673:684 the topology of Gpc1 with respect to the membrane 673:721 Therefore, we have studied Gpc1 using crystallography, small angle x-ray scattering, and chromatographic approaches to elucidate the composition, structure, and function of the N-glycans and the C terminus and also the topology of Gpc1 with respect to the membrane. 26203194 3 42 link N-linked 317:324 arg1 glycans 326:332 The two N-linked glycans 309:332 The two N-linked glycans 309:332 The two N-linked glycans and the C-terminal domain that attach the core protein to the cell membrane are not resolved in the Gpc1 crystal structure. 26203194 5 43 theme cell 1007:1010 arg1 surface 1012:1018 the cell surface 1003:1018 the cell surface 1003:1018 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 44 theme conserved 878:886 arg1 surface 855:861 a surface 853:861 a surface evolutionarily conserved 853:886 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 0 45 theme Human 68:72 arg1 Glypican-1 74:83 Human Glypican-1 68:83 Human Glypican-1 68:83 Structural Aspects of N-Glycosylations and the C-terminal Region in Human Glypican-1. 26203194 5 46 theme core 802:805 arg1 protein 807:813 the core protein 798:813 the core protein transverse to the membrane 798:840 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 1 47 theme several 155:161 arg1 pathways 192:199 several important cellular signaling pathways 155:199 several important cellular signaling pathways 155:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 26203194 5 48 theme transverse 815:824 arg1 protein 807:813 the core protein 798:813 the core protein transverse to the membrane 798:840 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 49 theme sulfate 1061:1067 arg1 substitution 1069:1080 heparan sulfate substitution 1053:1080 heparan sulfate substitution in the Golgi apparatus 1053:1103 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 5 50 theme signaling 954:962 arg1 molecules 964:972 signaling molecules 954:972 signaling molecules 954:972 The C terminus is shown to be highly flexible in solution, but it orients the core protein transverse to the membrane, directing a surface evolutionarily conserved in Gpc1 orthologs toward the membrane, where it may interact with signaling molecules and/or membrane receptors on the cell surface, or even the enzymes involved in heparan sulfate substitution in the Golgi apparatus. 26203194 1 51 theme important 163:171 arg1 pathways 192:199 several important cellular signaling pathways 155:199 several important cellular signaling pathways 155:199 Glypicans are multifunctional cell surface proteoglycans involved in several important cellular signaling pathways. 24295789 2 0 theme molecular 448:456 arg1 mass 458:461 molecular mass 448:461 molecular mass of ∼66,000 and ∼44,000 Da 448:487 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 3 1 theme interaction 685:695 arg1 chromatography 704:717 zwitterionic hydrophilic interaction liquid chromatography 660:717 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 5 2 theme sequence 1189:1196 arg1 analysis 1198:1205 the N-glycan sequence analysis 1176:1205 the N-glycan sequence analysis 1176:1205 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 6 3 theme acidic 1467:1472 arg1 α-mannosidases 1474:1487 all the acidic α-mannosidases 1459:1487 all the acidic α-mannosidases reported so far in eukaryotes 1459:1517 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 3 4 theme solid 719:723 arg1 separation 748:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 5 theme low-energy 860:869 arg1 dissociation 889:900 low-energy collision-induced dissociation 860:900 low-energy collision-induced dissociation 860:900 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 2 6 theme purified 362:369 arg1 enzyme 371:376 the purified enzyme 358:376 the purified enzyme 358:376 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 1 7 used used 298:301 arg2 properties 268:277 its biochemical properties 252:277 its biochemical properties 252:277 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 2 8 theme present 330:336 arg1 study 338:342 the present study 326:342 the present study 326:342 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 6 9 from present 1448:1454 arg1 α-mannosidases 1474:1487 all the acidic α-mannosidases 1459:1487 all the acidic α-mannosidases reported so far in eukaryotes 1459:1517 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 3 10 theme collision-induced 871:887 arg1 dissociation 889:900 low-energy collision-induced dissociation 860:900 low-energy collision-induced dissociation 860:900 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 0 11 theme glycomics 90:98 arg1 tool 100:103 a valuable glycomics tool 79:103 a valuable glycomics tool 79:103 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 12 theme zwitterionic 660:671 arg1 chromatography 704:717 zwitterionic hydrophilic interaction liquid chromatography 660:717 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 2 13 dep subunits 438:445 arg1 mass 458:461 molecular mass 448:461 molecular mass of ∼66,000 and ∼44,000 Da 448:487 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 2 13 dep subunits 438:445 arg1 both 423:426 both 423:426 both 423:426 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 0 14 gly N-glycosylation 51:65 arg1 tool 100:103 a valuable glycomics tool 79:103 a valuable glycomics tool 79:103 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 15 theme spectrometry 843:854 arg1 experiments 902:912 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments 784:912 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments 784:912 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 0 16 theme valuable 81:88 arg1 tool 100:103 a valuable glycomics tool 79:103 a valuable glycomics tool 79:103 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 1 17 contain contain 145:151 arg2 proteins 184:191 several biologically important proteins 153:191 several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis 153:320 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 1 17 contain contain 145:151 arg1 Canavalia 117:125 Canavalia 117:125 Canavalia 117:125 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 1 17 contain contain 145:151 arg1 seeds 139:143 Jack bean (Canavalia ensiformis) seeds 106:143 Jack bean (Canavalia ensiformis) seeds 106:143 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 7 18 theme paucimannose 1541:1552 arg1 type 1554:1557 a truncated paucimannose type 1529:1557 a truncated paucimannose type 1529:1557 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 1 19 dep Canavalia 117:125 arg1 ensiformis 127:136 Canavalia ensiformis 117:136 Canavalia ensiformis 117:136 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 4 20 theme site 1066:1069 arg1 specificities 1071:1083 site specificities 1066:1083 site specificities 1066:1083 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 3 21 gly N-glycopeptides 583:597 arg2 N-glycopeptides 583:597 N-glycopeptides 583:597 N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 583:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 4 22 theme complete 1026:1033 arg1 structures 1044:1053 complete N-glycan structures 1026:1053 complete N-glycan structures 1026:1053 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 3 23 theme phase 725:729 arg1 separation 748:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 5 24 theme hexoses 1307:1313 arg1 number 1297:1302 number 1297:1302 number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 1297:1352 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 6 25 from α-mannosidases 1474:1487 arg1 present 1448:1454 present 1448:1454 present 1448:1454 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 1 26 theme several 153:159 arg1 proteins 184:191 several biologically important proteins 153:191 several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis 153:320 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 0 27 theme tool 100:103 arg1 sequencing 36:45 amino acid sequencing 25:45 amino acid sequencing 25:45 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 0 27 theme tool 100:103 arg1 analysis 67:74 N-glycosylation analysis 51:74 N-glycosylation analysis 51:74 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 28 theme liquid 697:702 arg1 chromatography 704:717 zwitterionic hydrophilic interaction liquid chromatography 660:717 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 29 dep de 534:535 arg1 novo 537:540 novo 537:540 novo 537:540 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 0 30 theme bean 5:8 arg1 α-mannosidase 10:22 Jack bean α-mannosidase 0:22 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 0:104 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 31 theme chromatography 704:717 arg1 separation 748:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 4 32 theme De 915:916 arg1 sequencing 923:932 De novo sequencing 915:932 De novo sequencing 915:932 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 4 33 theme subunits 1013:1020 arg1 organization 983:994 the disulfide linkage organization 961:994 the disulfide linkage organization 961:994 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 4 33 theme subunits 1013:1020 arg1 structures 1044:1053 complete N-glycan structures 1026:1053 complete N-glycan structures 1026:1053 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 4 33 theme subunits 1013:1020 arg1 intersection 997:1008 intersection 997:1008 intersection of subunits 997:1020 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 1 34 theme important 174:182 arg1 proteins 184:191 several biologically important proteins 153:191 several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis 153:320 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 4 35 dep De 915:916 arg1 novo 918:921 novo 918:921 novo 918:921 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 1 36 theme biochemical 256:266 arg1 properties 268:277 its biochemical properties 252:277 its biochemical properties 252:277 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 6 37 located present 1448:1454 arg2 site 1412:1415 This site 1407:1415 This site with two proximal cysteines 1407:1443 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 6 37 located present 1448:1454 arg1 α-mannosidases 1474:1487 all the acidic α-mannosidases 1459:1487 all the acidic α-mannosidases reported so far in eukaryotes 1459:1517 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 3 38 theme nanoelectrospray 784:799 arg1 spectrometry 843:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 2 39 theme acid 400:403 arg1 sequence 405:412 its amino acid sequence 390:412 its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety 390:523 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 2 40 theme amino 394:398 arg1 sequence 405:412 its amino acid sequence 390:412 its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety 390:523 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 3 41 theme extraction-based 731:746 arg1 separation 748:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 4 42 theme linkage 975:981 arg1 organization 983:994 the disulfide linkage organization 961:994 the disulfide linkage organization 961:994 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 5 43 gly N-glycosylation 1385:1399 arg2 site 1401:1404 an evolutionarily conserved N-glycosylation site 1357:1404 an evolutionarily conserved N-glycosylation site 1357:1404 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 6 44 with site 1412:1415 arg1 cysteines 1435:1443 two proximal cysteines 1422:1443 two proximal cysteines 1422:1443 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 0 45 theme acid 31:34 arg1 sequencing 36:45 amino acid sequencing 25:45 amino acid sequencing 25:45 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 5 46 theme hydrolase 1152:1160 arg1 family 1162:1167 glycosyl hydrolase family 38 1143:1170 glycosyl hydrolase family 38 1143:1170 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 47 theme primary 1090:1096 arg1 sequence 1098:1105 The primary sequence 1086:1105 The primary sequence 1086:1105 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 0 48 theme amino 25:29 arg1 sequencing 36:45 amino acid sequencing 25:45 amino acid sequencing 25:45 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 2 49 theme known 432:436 arg1 subunits 438:445 the known subunits 428:445 the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) 428:488 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 3 50 theme N-glycopeptides 583:597 arg1 sequencing 542:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 50 theme N-glycopeptides 583:597 arg1 elucidation 568:578 structural elucidation 557:578 structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 557:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 5 51 theme glycosyl 1143:1150 arg1 family 1162:1167 glycosyl hydrolase family 38 1143:1170 glycosyl hydrolase family 38 1143:1170 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 2 52 theme Da 486:487 arg1 mass 458:461 molecular mass 448:461 molecular mass of ∼66,000 and ∼44,000 Da 448:487 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 4 53 theme new 943:945 arg1 insights 947:954 new insights 943:954 new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures 943:1053 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 4 54 theme N-glycan 1035:1042 arg1 structures 1044:1053 complete N-glycan structures 1026:1053 complete N-glycan structures 1026:1053 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 3 55 theme Peptide 526:532 arg1 sequencing 542:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 5 56 theme N-glycan 1180:1187 arg1 analysis 1198:1205 the N-glycan sequence analysis 1176:1205 the N-glycan sequence analysis 1176:1205 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 57 theme high-mannose 1216:1227 arg1 oligosaccharides 1229:1244 high-mannose oligosaccharides 1216:1244 high-mannose oligosaccharides 1216:1244 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 6 58 theme proximal 1426:1433 arg1 cysteines 1435:1443 two proximal cysteines 1422:1443 two proximal cysteines 1422:1443 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 7 59 theme terminal 1588:1595 arg1 mannose 1601:1607 terminal two mannose 1588:1607 terminal two mannose 1588:1607 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 59 theme terminal 1588:1595 arg1 GlcNAc2 1619:1625 Man1(Xyl)GlcNAc2 1610:1625 Man1(Xyl)GlcNAc2 (Fuc) 1610:1631 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 60 theme Man1 1610:1613 arg1 Fuc 1628:1630 Fuc 1628:1630 Fuc 1628:1630 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 60 theme Man1 1610:1613 arg1 GlcNAc2 1619:1625 Man1(Xyl)GlcNAc2 1610:1625 Man1(Xyl)GlcNAc2 (Fuc) 1610:1631 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 60 theme Man1 1610:1613 arg1 mannose 1601:1607 terminal two mannose 1588:1607 terminal two mannose 1588:1607 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 3 61 theme de 534:535 arg1 sequencing 542:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing 526:551 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 7 62 theme truncated 1531:1539 arg1 type 1554:1557 a truncated paucimannose type 1529:1557 a truncated paucimannose type 1529:1557 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 1 63 theme bean 111:114 arg1 Canavalia 117:125 Canavalia 117:125 Canavalia 117:125 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 1 63 theme bean 111:114 arg1 seeds 139:143 Jack bean (Canavalia ensiformis) seeds 106:143 Jack bean (Canavalia ensiformis) seeds 106:143 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 5 64 theme conserved 1375:1383 arg1 site 1401:1404 an evolutionarily conserved N-glycosylation site 1357:1404 an evolutionarily conserved N-glycosylation site 1357:1404 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 1 65 theme glycan 306:311 arg1 analysis 313:320 glycan analysis 306:320 glycan analysis 306:320 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 4 66 theme disulfide 965:973 arg1 organization 983:994 the disulfide linkage organization 961:994 the disulfide linkage organization 961:994 De novo sequencing provided new insights into the disulfide linkage organization, intersection of subunits and complete N-glycan structures along with site specificities. 24295789 5 67 dep hexoses 1307:1313 arg1 Glc1Man9GlcNAc2 1338:1352 Glc1Man9GlcNAc2 1338:1352 Glc1Man9GlcNAc2 1338:1352 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 67 dep hexoses 1307:1313 arg1 hexoses 1307:1313 hexoses 1307:1313 hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 1307:1352 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 67 dep hexoses 1307:1313 arg1 Man8-9GlcNAc2 1320:1332 Man8-9GlcNAc2 1320:1332 Man8-9GlcNAc2 1320:1332 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 67 dep hexoses 1307:1313 arg1 viz 1315:1317 viz 1315:1317 viz 1315:1317 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 5 68 theme N-glycosylation 1385:1399 arg1 site 1401:1404 an evolutionarily conserved N-glycosylation site 1357:1404 an evolutionarily conserved N-glycosylation site 1357:1404 The primary sequence suggests that the enzyme belongs to glycosyl hydrolase family 38 and the N-glycan sequence analysis revealed high-mannose oligosaccharides, which were found to be heterogeneous with varying number of hexoses viz, Man8-9GlcNAc2 and Glc1Man9GlcNAc2 in an evolutionarily conserved N-glycosylation site. 24295789 1 69 theme EC 220:221 arg1 α-mannosidase 205:217 α-mannosidase 205:217 α-mannosidase (EC 3.2.1.24) 205:231 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 1 69 theme EC 220:221 arg1 3.2.1.24 223:230 EC 3.2.1.24 220:230 EC 3.2.1.24 220:230 Jack bean (Canavalia ensiformis) seeds contain several biologically important proteins among which α-mannosidase (EC 3.2.1.24) has been purified, its biochemical properties studied and widely used in glycan analysis. 24295789 6 70 attach present 1448:1454 arg1 α-mannosidases 1474:1487 all the acidic α-mannosidases 1459:1487 all the acidic α-mannosidases reported so far in eukaryotes 1459:1517 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 6 70 attach present 1448:1454 arg2 site 1412:1415 This site 1407:1415 This site with two proximal cysteines 1407:1443 This site with two proximal cysteines is present in all the acidic α-mannosidases reported so far in eukaryotes. 24295789 0 71 dep α-mannosidase 10:22 arg1 sequencing 36:45 amino acid sequencing 25:45 amino acid sequencing 25:45 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 0 71 dep α-mannosidase 10:22 arg1 analysis 67:74 N-glycosylation analysis 51:74 N-glycosylation analysis 51:74 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 72 theme dissociation 889:900 arg1 experiments 902:912 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments 784:912 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments 784:912 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 73 theme mass 838:841 arg1 spectrometry 843:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 74 theme proteolytic 629:639 arg1 digestion 641:649 proteolytic digestion 629:649 proteolytic digestion 629:649 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 2 75 used used 353:356 arg2 we 345:346 we 345:346 we 345:346 In the present study, we have used the purified enzyme and derived its amino acid sequence covering both the known subunits (molecular mass of ∼66,000 and ∼44,000 Da) hitherto not known in its entirety. 24295789 3 76 theme experiments 902:912 arg1 use 777:779 use 777:779 use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments 777:912 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 77 theme ionization 801:810 arg1 spectrometry 843:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 0 78 theme N-glycosylation 51:65 arg1 analysis 67:74 N-glycosylation analysis 51:74 N-glycosylation analysis 51:74 Jack bean α-mannosidase: amino acid sequencing and N-glycosylation analysis of a valuable glycomics tool. 24295789 3 79 theme hydrophilic 673:683 arg1 chromatography 704:717 zwitterionic hydrophilic interaction liquid chromatography 660:717 zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 660:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 80 theme structural 557:566 arg1 elucidation 568:578 structural elucidation 557:578 structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation 557:757 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 7 81 theme Xyl 1615:1617 arg1 Fuc 1628:1630 Fuc 1628:1630 Fuc 1628:1630 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 81 theme Xyl 1615:1617 arg1 GlcNAc2 1619:1625 Man1(Xyl)GlcNAc2 1610:1625 Man1(Xyl)GlcNAc2 (Fuc) 1610:1631 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 7 81 theme Xyl 1615:1617 arg1 mannose 1601:1607 terminal two mannose 1588:1607 terminal two mannose 1588:1607 Further, a truncated paucimannose type was identified to be lacking terminal two mannose, Man1(Xyl)GlcNAc2 (Fuc). 24295789 3 82 theme time-of-flight 823:836 arg1 spectrometry 843:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 24295789 3 83 theme quadrupole 812:821 arg1 spectrometry 843:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 nanoelectrospray ionization quadrupole time-of-flight mass spectrometry 784:854 Peptide de novo sequencing and structural elucidation of N-glycopeptides obtained either directly from proteolytic digestion or after zwitterionic hydrophilic interaction liquid chromatography solid phase extraction-based separation were performed by use of nanoelectrospray ionization quadrupole time-of-flight mass spectrometry and low-energy collision-induced dissociation experiments. 27249581 0 0 theme Carbohydrate 86:97 arg1 Structure 69:77 the Structure 65:77 the Structure of the Carbohydrate 65:97 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. 27249581 3 1 theme interactions 357:368 arg1 strengths 329:337 the relative strengths 316:337 the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host 316:483 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 10 2 theme recent 1650:1655 arg1 literature 1657:1666 recent literature 1650:1666 recent literature regarding the contribution of electrostatics to CH-π interactions 1650:1732 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 7 3 theme strongest 1120:1128 arg1 interactions 1145:1156 the strongest sugar-aromatic interactions 1116:1156 the strongest sugar-aromatic interactions 1116:1156 Galactose seems to make the weakest and allose the strongest sugar-aromatic interactions, with glucose, N-acetylglucosamine (GlcNAc) and mannose in between. 27249581 4 4 theme face-to-face 613:624 arg1 interactions 650:661 face-to-face monosaccharide-aromatic interactions 613:661 face-to-face monosaccharide-aromatic interactions 613:661 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 5 theme structural 530:539 arg1 motif 541:545 a structural motif 528:545 a structural motif found in the reverse turns of some N-glycoproteins 528:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 5 theme structural 530:539 arg1 sequon 520:525 the enhanced aromatic sequon 498:525 the enhanced aromatic sequon 498:525 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 0 6 from Dependence 4:13 arg1 Structure 69:77 the Structure 65:77 the Structure of the Carbohydrate 65:97 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. 27249581 10 7 link N-linked 1857:1864 arg1 glycans 1866:1872 N-linked glycans 1857:1872 N-linked glycans 1857:1872 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 9 8 from strength 1520:1527 arg1 context 1570:1576 the context 1566:1576 the context of our N-glycoprotein host 1566:1603 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 7 9 theme sugar-aromatic 1130:1143 arg1 interactions 1145:1156 the strongest sugar-aromatic interactions 1116:1156 the strongest sugar-aromatic interactions 1116:1156 Galactose seems to make the weakest and allose the strongest sugar-aromatic interactions, with glucose, N-acetylglucosamine (GlcNAc) and mannose in between. 27249581 10 10 from glycoproteins 1890:1902 arg1 eukaryotes 1907:1916 eukaryotes 1907:1916 eukaryotes 1907:1916 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 3 11 gly glycosylation 440:452 arg2 site 454:457 the glycosylation site 436:457 the glycosylation site 436:457 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 6 12 from changes 968:974 arg1 identity 1003:1010 identity 1003:1010 identity 1003:1010 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 6 12 from changes 968:974 arg1 stereochemistry 983:997 stereochemistry 983:997 stereochemistry 983:997 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 5 13 theme folding 718:724 arg1 energetics 726:735 the folding energetics 714:735 the folding energetics 714:735 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 5 14 theme protein 666:672 arg1 host 674:677 A protein host 664:677 A protein host 664:677 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 4 15 theme monosaccharide-aromatic 626:648 arg1 interactions 650:661 face-to-face monosaccharide-aromatic interactions 613:661 face-to-face monosaccharide-aromatic interactions 613:661 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 6 16 from stereochemistry 983:997 arg1 rings 1048:1052 the pyranose rings 1035:1052 the pyranose rings of the sugars 1035:1066 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 4 17 theme aromatic 511:518 arg1 motif 541:545 a structural motif 528:545 a structural motif found in the reverse turns of some N-glycoproteins 528:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 17 theme aromatic 511:518 arg1 sequon 520:525 the enhanced aromatic sequon 498:525 the enhanced aromatic sequon 498:525 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 1 18 from biology 166:172 arg1 ubiquitous 152:161 ubiquitous 152:161 ubiquitous 152:161 Interactions between proteins and carbohydrates are ubiquitous in biology. 27249581 9 19 theme N-glycoprotein 1585:1598 arg1 host 1600:1603 our N-glycoprotein host 1581:1603 our N-glycoprotein host 1581:1603 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 8 20 theme monosaccharide-containing 1272:1296 arg1 N-glycoproteins 1298:1312 the monosaccharide-containing N-glycoproteins 1268:1312 the monosaccharide-containing N-glycoproteins 1268:1312 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 9 21 theme host 1600:1603 arg1 context 1570:1576 the context 1566:1576 the context of our N-glycoprotein host 1566:1603 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 3 22 theme monosaccharides 390:404 arg1 series 380:385 a series 378:385 a series of monosaccharides 378:404 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 9 23 theme interaction 1551:1561 arg1 strength 1520:1527 the strength 1516:1527 the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host 1516:1603 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 8 24 theme several 1257:1263 arg1 structures 1243:1252 The NMR solution structures 1226:1252 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins 1226:1312 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 1 25 from ubiquitous 152:161 arg1 biology 166:172 biology 166:172 biology 166:172 Interactions between proteins and carbohydrates are ubiquitous in biology. 27249581 6 26 from substituents 1019:1030 arg1 rings 1048:1052 the pyranose rings 1035:1052 the pyranose rings of the sugars 1035:1066 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 10 27 theme GlcNAc 1814:1819 arg1 what 1751:1754 what 1751:1754 what 1751:1754 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 27 theme GlcNAc 1814:1819 arg1 monosaccharide 1828:1841 the monosaccharide 1824:1841 the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes 1824:1916 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 27 theme GlcNAc 1814:1819 arg1 conservation 1798:1809 the absolute conservation 1785:1809 the absolute conservation of GlcNAc 1785:1819 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 28 attach attached 1878:1885 arg2 glycans 1866:1872 N-linked glycans 1857:1872 N-linked glycans 1857:1872 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 28 attach attached 1878:1885 arg1 glycoproteins 1890:1902 glycoproteins 1890:1902 glycoproteins in eukaryotes 1890:1916 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 4 29 theme N-glycoproteins 582:596 arg1 turns 568:572 the reverse turns 556:572 the reverse turns of some N-glycoproteins 556:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 8 30 theme NMR 1230:1232 arg1 structures 1243:1252 The NMR solution structures 1226:1252 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins 1226:1312 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 3 31 theme N-glycoprotein 465:478 arg1 host 480:483 an N-glycoprotein host 462:483 an N-glycoprotein host 462:483 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 3 32 theme aromatic 413:420 arg1 ring 422:425 an aromatic ring 410:425 an aromatic ring close to the glycosylation site 410:457 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 8 33 theme interaction 1432:1442 arg1 energies 1444:1451 the monosaccharide-aromatic interaction energies 1404:1451 the monosaccharide-aromatic interaction energies 1404:1451 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 10 34 gly glycoproteins 1890:1902 arg1 glycoproteins 1890:1902 glycoproteins 1890:1902 glycoproteins in eukaryotes 1890:1916 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 8 35 gly N-glycoproteins 1298:1312 arg1 N-glycoproteins 1298:1312 the monosaccharide-containing N-glycoproteins 1268:1312 the monosaccharide-containing N-glycoproteins 1268:1312 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 8 36 theme N-glycoproteins 1298:1312 arg1 several 1257:1263 several 1257:1263 several 1257:1263 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 8 37 theme similarities 1367:1378 arg1 origins 1352:1358 the origins 1348:1358 the origins of the similarities and differences between the monosaccharide-aromatic interaction energies 1348:1451 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 3 38 theme ring 422:425 arg1 strengths 329:337 the relative strengths 316:337 the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host 316:483 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 6 39 from rings 1048:1052 arg1 identity 1003:1010 identity 1003:1010 identity 1003:1010 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 6 39 from rings 1048:1052 arg1 stereochemistry 983:997 stereochemistry 983:997 stereochemistry 983:997 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 0 40 theme Strengths 52:60 arg1 Dependence 4:13 The Dependence 0:13 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate 0:97 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. 27249581 5 41 theme monosaccharide 760:773 arg1 identity 744:751 the identity 740:751 the identity of the monosaccharide 740:773 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 10 42 theme N-linked 1857:1864 arg1 glycans 1866:1872 N-linked glycans 1857:1872 N-linked glycans 1857:1872 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 8 43 theme differences 1384:1394 arg1 origins 1352:1358 the origins 1348:1358 the origins of the similarities and differences between the monosaccharide-aromatic interaction energies 1348:1451 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 0 44 theme Interaction 40:50 arg1 Strengths 52:60 Carbohydrate-Aromatic Interaction Strengths 18:60 Carbohydrate-Aromatic Interaction Strengths 18:60 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. 27249581 4 45 theme reverse 560:566 arg1 turns 568:572 the reverse turns 556:572 the reverse turns of some N-glycoproteins 556:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 5 46 theme carbohydrate-aromatic 832:852 arg1 interaction 854:864 the carbohydrate-aromatic interaction 828:864 the carbohydrate-aromatic interaction 828:864 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 4 47 theme enhanced 502:509 arg1 motif 541:545 a structural motif 528:545 a structural motif found in the reverse turns of some N-glycoproteins 528:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 47 theme enhanced 502:509 arg1 sequon 520:525 the enhanced aromatic sequon 498:525 the enhanced aromatic sequon 498:525 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 0 48 theme Carbohydrate-Aromatic 18:38 arg1 Strengths 52:60 Carbohydrate-Aromatic Interaction Strengths 18:60 Carbohydrate-Aromatic Interaction Strengths 18:60 The Dependence of Carbohydrate-Aromatic Interaction Strengths on the Structure of the Carbohydrate. 27249581 10 49 theme CH-π 1716:1719 arg1 interactions 1721:1732 CH-π interactions 1716:1732 CH-π interactions 1716:1732 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 5 50 theme interaction 854:864 arg1 strength 816:823 the strength 812:823 the strength of the carbohydrate-aromatic interaction 812:864 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 6 51 theme pyranose 1039:1046 arg1 rings 1048:1052 the pyranose rings 1035:1052 the pyranose rings of the sugars 1035:1066 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 5 52 theme energetics 726:735 arg1 dependence 700:709 the dependence 696:709 the dependence of the folding energetics on the identity of the monosaccharide 696:773 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 9 53 theme monosaccharides 1476:1490 arg1 Peracetylation 1454:1467 Peracetylation 1454:1467 Peracetylation of the monosaccharides 1454:1490 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 4 54 gly N-glycoproteins 582:596 arg1 N-glycoproteins 582:596 some N-glycoproteins 577:596 some N-glycoproteins 577:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 3 55 theme glycosylation 440:452 arg1 site 454:457 the glycosylation site 436:457 the glycosylation site 436:457 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 5 56 from dependence 700:709 arg1 identity 744:751 the identity 740:751 the identity of the monosaccharide 740:773 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 3 57 theme intramolecular 342:355 arg1 interactions 357:368 intramolecular interactions 342:368 intramolecular interactions between a series of monosaccharides 342:404 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 10 58 theme electrostatics 1698:1711 arg1 contribution 1682:1693 the contribution 1678:1693 the contribution of electrostatics to CH-π interactions 1678:1732 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 6 59 theme substituents 1019:1030 arg1 identity 1003:1010 identity 1003:1010 identity 1003:1010 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 6 59 theme substituents 1019:1030 arg1 stereochemistry 983:997 stereochemistry 983:997 stereochemistry 983:997 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 3 60 theme relative 320:327 arg1 strengths 329:337 the relative strengths 316:337 the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host 316:483 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 6 61 theme interaction 919:929 arg1 strengths 931:939 the carbohydrate-aromatic interaction strengths 893:939 the carbohydrate-aromatic interaction strengths 893:939 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 5 62 used used 683:686 arg2 host 674:677 A protein host 664:677 A protein host 664:677 A protein host was used because the dependence of the folding energetics on the identity of the monosaccharide can be accurately measured to assess the strength of the carbohydrate-aromatic interaction. 27249581 4 63 located found 547:551 arg1 turns 568:572 the reverse turns 556:572 the reverse turns of some N-glycoproteins 556:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 63 located found 547:551 arg2 motif 541:545 a structural motif 528:545 a structural motif found in the reverse turns of some N-glycoproteins 528:596 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 4 63 located found 547:551 arg2 sequon 520:525 the enhanced aromatic sequon 498:525 the enhanced aromatic sequon 498:525 We employed the enhanced aromatic sequon, a structural motif found in the reverse turns of some N-glycoproteins, to facilitate face-to-face monosaccharide-aromatic interactions. 27249581 9 64 theme sugar-aromatic 1536:1549 arg1 interaction 1551:1561 the sugar-aromatic interaction 1532:1561 the sugar-aromatic interaction 1532:1561 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 10 65 theme literature 1657:1666 arg1 light 1641:1645 light 1641:1645 light of recent literature regarding the contribution of electrostatics to CH-π interactions 1641:1732 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 6 66 theme carbohydrate-aromatic 897:917 arg1 strengths 931:939 the carbohydrate-aromatic interaction strengths 893:939 the carbohydrate-aromatic interaction strengths 893:939 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 8 67 theme monosaccharide-aromatic 1408:1430 arg1 energies 1444:1451 the monosaccharide-aromatic interaction energies 1404:1451 the monosaccharide-aromatic interaction energies 1404:1451 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27249581 3 68 from strengths 329:337 arg1 host 480:483 an N-glycoprotein host 462:483 an N-glycoprotein host 462:483 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 6 69 from identity 1003:1010 arg1 rings 1048:1052 the pyranose rings 1035:1052 the pyranose rings of the sugars 1035:1066 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 3 70 gly N-glycoprotein 465:478 arg1 N-glycoprotein 465:478 N-glycoprotein 465:478 N-glycoprotein 465:478 Herein, we have determined the relative strengths of intramolecular interactions between a series of monosaccharides and an aromatic ring close to the glycosylation site in an N-glycoprotein host. 27249581 9 71 gly N-glycoprotein 1585:1598 arg1 N-glycoprotein 1585:1598 N-glycoprotein 1585:1598 N-glycoprotein 1585:1598 Peracetylation of the monosaccharides substantially increases the strength of the sugar-aromatic interaction in the context of our N-glycoprotein host. 27249581 6 72 dep stereochemistry 983:997 arg1 the 979:981 the 979:981 the 979:981 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 6 73 theme sugars 1061:1066 arg1 rings 1048:1052 the pyranose rings 1035:1052 the pyranose rings of the sugars 1035:1066 Our data demonstrate that the carbohydrate-aromatic interaction strengths are moderately affected by changes in the stereochemistry and identity of the substituents on the pyranose rings of the sugars. 27249581 10 74 theme absolute 1789:1796 arg1 what 1751:1754 what 1751:1754 what 1751:1754 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 74 theme absolute 1789:1796 arg1 monosaccharide 1828:1841 the monosaccharide 1824:1841 the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes 1824:1916 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 10 74 theme absolute 1789:1796 arg1 conservation 1798:1809 the absolute conservation 1785:1809 the absolute conservation of GlcNAc 1785:1819 Finally, we discuss our results in light of recent literature regarding the contribution of electrostatics to CH-π interactions and speculate on what our observations imply about the absolute conservation of GlcNAc as the monosaccharide through which N-linked glycans are attached to glycoproteins in eukaryotes. 27249581 8 75 theme solution 1234:1241 arg1 structures 1243:1252 The NMR solution structures 1226:1252 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins 1226:1312 The NMR solution structures of several of the monosaccharide-containing N-glycoproteins were solved to further understand the origins of the similarities and differences between the monosaccharide-aromatic interaction energies. 27054587 6 0 theme them 964:967 arg1 β-1,4-galactosyltransferase 970:996 β-1,4-galactosyltransferase 970:996 β-1,4-galactosyltransferase (β4Gal-T4) 970:1007 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 6 0 theme them 964:967 arg1 them 964:967 them 964:967 them 964:967 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 6 0 theme them 964:967 arg1 six 957:959 six 957:959 six 957:959 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 7 1 theme sialyl-Lewis 1165:1176 arg1 X 1178:1178 sialyl-Lewis X 1165:1178 sialyl-Lewis X 1165:1178 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 5 2 theme Simple 741:746 arg1 measures 764:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures were used to characterise the resulting reaction networks. 27054587 1 3 theme post-translational 161:178 arg1 glycosylation 131:143 O-linked glycosylation 122:143 O-linked glycosylation 122:143 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 3 theme post-translational 161:178 arg1 changes 216:222 changes 216:222 changes to which are important biomarkers of cancer 216:266 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 3 theme post-translational 161:178 arg1 modification 180:191 an important post-translational modification 148:191 an important post-translational modification of mucin-type protein 148:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 3 4 from activities 583:592 arg1 glycans 559:565 glycans 559:565 glycans from the enzyme activities 559:592 Software for generating glycans from the enzyme activities is presented, and is also available online. 27054587 2 5 theme reactions 524:532 arg1 networks 495:502 networks 495:502 networks of enzyme-catalysed reactions 495:532 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 7 6 theme X 1178:1178 arg1 glycoforms 1201:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 6 7 theme each 885:888 arg1 knockouts 872:880 in-silico single-enzyme knockouts 848:880 in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis 848:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 7 8 theme X 1162:1162 arg1 glycoforms 1201:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 0 9 dep Knockouts 111:119 arg1 Response 92:99 Response 92:99 Response 92:99 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 5 10 theme resulting 803:811 arg1 networks 822:829 the resulting reaction networks 799:829 the resulting reaction networks 799:829 Simple graph-theoretic measures were used to characterise the resulting reaction networks. 27054587 2 11 link GalNAc-linked 370:382 arg1 oligosaccharides 384:399 GalNAc-linked oligosaccharides 370:399 GalNAc-linked oligosaccharides 370:399 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 2 12 theme enzymes 291:297 arg1 study 278:282 this study 273:282 this study of the enzymes of O-glycosylation 273:316 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 6 13 theme knockouts 872:880 arg1 study 839:843 a study 837:843 a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis 837:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 2 14 theme GalNAc-linked 370:382 arg1 oligosaccharides 384:399 GalNAc-linked oligosaccharides 370:399 GalNAc-linked oligosaccharides 370:399 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 4 15 from Poisson 722:728 arg1 nature 733:738 nature 733:738 nature 733:738 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 8 16 dep 98 1534:1535 arg1 to 1531:1532 to 1531:1532 to 1531:1532 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 6 17 theme single-enzyme 858:870 arg1 knockouts 872:880 in-silico single-enzyme knockouts 848:880 in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis 848:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 3 18 theme enzyme 576:581 arg1 activities 583:592 the enzyme activities 572:592 the enzyme activities 572:592 Software for generating glycans from the enzyme activities is presented, and is also available online. 27054587 0 19 theme Enzyme 104:109 arg1 Knockouts 111:119 Enzyme Knockouts 104:119 Enzyme Knockouts 104:119 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 6 20 theme O-glycan 934:941 arg1 biosynthesis 943:954 mucin O-glycan biosynthesis 928:954 mucin O-glycan biosynthesis 928:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 5 21 used used 778:781 arg2 measures 764:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures were used to characterise the resulting reaction networks. 27054587 6 22 theme in-silico 848:856 arg1 knockouts 872:880 in-silico single-enzyme knockouts 848:880 in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis 848:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 1 23 theme mucin-type 196:205 arg1 protein 207:213 mucin-type protein 196:213 mucin-type protein 196:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 0 24 theme Knowledge-Based 2:16 arg1 System 18:23 A Knowledge-Based System 0:23 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 0:120 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 0 25 theme Behaviour 79:87 arg1 Prediction 41:50 Prediction 41:50 Prediction 41:50 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 0 25 theme Behaviour 79:87 arg1 Display 29:35 Display 29:35 Display 29:35 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 5 26 theme reaction 813:820 arg1 networks 822:829 the resulting reaction networks 799:829 the resulting reaction networks 799:829 Simple graph-theoretic measures were used to characterise the resulting reaction networks. 27054587 8 27 theme O-glycans 1430:1438 arg1 O-glycans 1430:1438 244 experimentally determined mucin-type O-glycans 1389:1438 244 experimentally determined mucin-type O-glycans obtained from the literature 1389:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 27 theme O-glycans 1430:1438 arg1 set 1382:1384 A set 1380:1384 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature 1380:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 1 28 theme protein 207:213 arg1 glycosylation 131:143 O-linked glycosylation 122:143 O-linked glycosylation 122:143 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 28 theme protein 207:213 arg1 changes 216:222 changes 216:222 changes to which are important biomarkers of cancer 216:266 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 28 theme protein 207:213 arg1 modification 180:191 an important post-translational modification 148:191 an important post-translational modification of mucin-type protein 148:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 7 29 theme Y 1187:1187 arg1 glycoforms 1201:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 8 30 theme mucin-type 1419:1428 arg1 O-glycans 1430:1438 244 experimentally determined mucin-type O-glycans 1389:1438 244 experimentally determined mucin-type O-glycans obtained from the literature 1389:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 2 31 theme pattern-matching 453:468 arg1 algorithm 470:478 a pattern-matching algorithm 451:478 a pattern-matching algorithm that generates networks of enzyme-catalysed reactions 451:532 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 5 32 theme graph-theoretic 748:762 arg1 measures 764:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures 741:771 Simple graph-theoretic measures were used to characterise the resulting reaction networks. 27054587 7 33 dep β4Gal-T4 1142:1149 arg1 the 1127:1129 the 1127:1129 the 1127:1129 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 33 dep β4Gal-T4 1142:1149 arg1 absence 1131:1137 absence 1131:1137 absence 1131:1137 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 8 34 gly glycoforms 1612:1621 arg1 cell 1607:1610 engineered CHO cell glycoforms 1592:1621 engineered CHO cell glycoforms 1592:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 34 gly glycoforms 1612:1621 arg1 CHO 1603:1605 engineered CHO cell glycoforms 1592:1621 engineered CHO cell glycoforms 1592:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 4 35 theme enzyme-reaction 680:694 arg1 networks 696:703 the resulting enzyme-reaction networks 666:703 the resulting enzyme-reaction networks 666:703 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 8 36 theme common 1550:1555 arg1 structures 1557:1566 the most common structures 1541:1566 the most common structures obtained from human and engineered CHO cell glycoforms 1541:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 37 theme engineered 1592:1601 arg1 glycoforms 1612:1621 engineered CHO cell glycoforms 1592:1621 engineered CHO cell glycoforms 1592:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 38 theme determined 1408:1417 arg1 O-glycans 1430:1438 244 experimentally determined mucin-type O-glycans 1389:1438 244 experimentally determined mucin-type O-glycans obtained from the literature 1389:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 2 39 theme O-glycosylation 302:316 arg1 enzymes 291:297 the enzymes 287:297 the enzymes of O-glycosylation 287:316 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 4 40 theme resulting 670:678 arg1 networks 696:703 the resulting enzyme-reaction networks 666:703 the resulting enzyme-reaction networks 666:703 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 2 41 theme enzyme-catalysed 507:522 arg1 reactions 524:532 enzyme-catalysed reactions 507:532 enzyme-catalysed reactions 507:532 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 7 42 theme O-glycans 1343:1351 arg1 abundances 1329:1338 the relative abundances 1316:1338 the relative abundances of O-glycans expressing these epitopes 1316:1377 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 6 43 theme O-glycan 1100:1107 arg1 heterogeneity 1109:1121 O-glycan heterogeneity 1100:1121 O-glycan heterogeneity 1100:1121 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 8 44 theme cell 1607:1610 arg1 glycoforms 1612:1621 engineered CHO cell glycoforms 1592:1621 engineered CHO cell glycoforms 1592:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 7 45 theme Sda/Cad 1193:1199 arg1 glycoforms 1201:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms 1152:1210 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 2 46 theme graphical 421:429 arg1 interpretation 431:444 their graphical interpretation 415:444 their graphical interpretation 415:444 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 8 47 theme CHO 1603:1605 arg1 glycoforms 1612:1621 engineered CHO cell glycoforms 1592:1621 engineered CHO cell glycoforms 1592:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 7 48 theme N-acetylglucosaminyltransferases 1261:1292 arg1 knockouts 1244:1252 knockouts 1244:1252 knockouts 1244:1252 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 3 49 theme available 620:628 arg1 online 630:635 available online 620:635 available online 620:635 Software for generating glycans from the enzyme activities is presented, and is also available online. 27054587 7 50 theme relative 1320:1327 arg1 abundances 1329:1338 the relative abundances 1316:1338 the relative abundances of O-glycans expressing these epitopes 1316:1377 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 51 theme Lewis 1181:1185 arg1 Y 1187:1187 Lewis Y 1181:1187 Lewis Y 1181:1187 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 52 dep knockouts 1244:1252 arg1 contrast 1232:1239 contrast 1232:1239 contrast 1232:1239 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 53 theme Lewis 1156:1160 arg1 X 1162:1162 Lewis X 1156:1162 Lewis X 1156:1162 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 1 54 theme O-linked 122:129 arg1 glycosylation 131:143 O-linked glycosylation 122:143 O-linked glycosylation 122:143 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 54 theme O-linked 122:129 arg1 modification 180:191 an important post-translational modification 148:191 an important post-translational modification of mucin-type protein 148:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 6 55 theme mucin 928:932 arg1 biosynthesis 943:954 mucin O-glycan biosynthesis 928:954 mucin O-glycan biosynthesis 928:954 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 2 56 theme shorthand 334:342 arg1 notation 344:351 a shorthand notation 332:351 a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions 332:532 For this study of the enzymes of O-glycosylation, we developed a shorthand notation for representing GalNAc-linked oligosaccharides, a method for their graphical interpretation, and a pattern-matching algorithm that generates networks of enzyme-catalysed reactions. 27054587 1 57 theme important 237:245 arg1 biomarkers 247:256 important biomarkers 237:256 important biomarkers of cancer 237:266 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 58 link O-linked 122:129 arg1 glycosylation 131:143 O-linked glycosylation 122:143 O-linked glycosylation 122:143 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 58 link O-linked 122:129 arg1 modification 180:191 an important post-translational modification 148:191 an important post-translational modification of mucin-type protein 148:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 8 59 used used 1473:1476 arg2 set 1382:1384 A set 1380:1384 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature 1380:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 59 used used 1473:1476 arg2 O-glycans 1430:1438 244 experimentally determined mucin-type O-glycans 1389:1438 244 experimentally determined mucin-type O-glycans obtained from the literature 1389:1467 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 4 60 theme degree 642:647 arg1 distributions 649:661 The degree distributions 638:661 The degree distributions of the resulting enzyme-reaction networks 638:703 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 4 60 theme degree 642:647 arg1 Poisson 722:728 Poisson 722:728 Poisson 722:728 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 0 61 theme Network 71:77 arg1 Behaviour 79:87 O-Glycosylation Network Behaviour 55:87 O-Glycosylation Network Behaviour 55:87 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 4 62 theme networks 696:703 arg1 distributions 649:661 The degree distributions 638:661 The degree distributions of the resulting enzyme-reaction networks 638:703 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 4 62 theme networks 696:703 arg1 Poisson 722:728 Poisson 722:728 Poisson 722:728 The degree distributions of the resulting enzyme-reaction networks were found to be Poisson in nature. 27054587 7 63 gly glycoforms 1201:1210 arg1 Y 1187:1187 Lewis Y 1181:1187 Lewis Y 1181:1187 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 63 gly glycoforms 1201:1210 arg1 X 1162:1162 Lewis X 1156:1162 Lewis X 1156:1162 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 63 gly glycoforms 1201:1210 arg1 Sda/Cad 1193:1199 Sda/Cad 1193:1199 Sda/Cad 1193:1199 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 7 63 gly glycoforms 1201:1210 arg1 X 1178:1178 sialyl-Lewis X 1165:1178 sialyl-Lewis X 1165:1178 In the absence of β4Gal-T4, all Lewis X, sialyl-Lewis X, Lewis Y and Sda/Cad glycoforms were eliminated, in contrast to knockouts of the N-acetylglucosaminyltransferases, which did not affect the relative abundances of O-glycans expressing these epitopes. 27054587 8 64 theme structures 1557:1566 arg1 structures 1557:1566 the most common structures 1541:1566 the most common structures obtained from human and engineered CHO cell glycoforms 1541:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 8 64 theme structures 1557:1566 arg1 % 1536:1536 up to 98% 1528:1536 up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms 1528:1621 A set of 244 experimentally determined mucin-type O-glycans obtained from the literature was used to validate the method, which was able to predict up to 98% of the most common structures obtained from human and engineered CHO cell glycoforms. 27054587 6 65 theme dominant 1071:1078 arg1 role 1080:1083 the dominant role 1067:1083 the dominant role 1067:1083 From a study of in-silico single-enzyme knockouts of each of 25 enzymes known to be involved in mucin O-glycan biosynthesis, six of them, β-1,4-galactosyltransferase (β4Gal-T4), four glycosyltransferases and one sulfotransferase, play the dominant role in determining O-glycan heterogeneity. 27054587 0 66 theme O-Glycosylation 55:69 arg1 Behaviour 79:87 O-Glycosylation Network Behaviour 55:87 O-Glycosylation Network Behaviour 55:87 A Knowledge-Based System for Display and Prediction of O-Glycosylation Network Behaviour in Response to Enzyme Knockouts. 27054587 1 67 theme important 151:159 arg1 glycosylation 131:143 O-linked glycosylation 122:143 O-linked glycosylation 122:143 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 67 theme important 151:159 arg1 changes 216:222 changes 216:222 changes to which are important biomarkers of cancer 216:266 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 67 theme important 151:159 arg1 modification 180:191 an important post-translational modification 148:191 an important post-translational modification of mucin-type protein 148:213 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 27054587 1 68 theme cancer 261:266 arg1 biomarkers 247:256 important biomarkers 237:256 important biomarkers of cancer 237:266 O-linked glycosylation is an important post-translational modification of mucin-type protein, changes to which are important biomarkers of cancer. 28371718 11 0 dep originator 1743:1752 arg1 biodrug 1768:1774 biodrug 1768:1774 biodrug 1768:1774 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 6 1 theme Intact 898:903 arg1 analysis 910:917 Intact mass analysis 898:917 Intact mass analysis 898:917 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 6 1 theme Intact 898:903 arg1 approach 930:937 middle-up approach 920:937 middle-up approach 920:937 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 1 2 theme due 167:169 arg1 market 160:165 the pharmaceuticals market 140:165 the pharmaceuticals market due to their high target selectivity in different diseases 140:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 11 3 theme functional 1591:1600 arg1 properties 1602:1611 functional properties 1591:1611 functional properties 1591:1611 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 5 4 theme post-translational 790:807 arg1 PTMs 824:827 PTMs 824:827 PTMs 824:827 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 5 4 theme post-translational 790:807 arg1 modifications 809:821 post-translational modifications 790:821 post-translational modifications (PTMs) 790:828 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 4 5 dep authorities 528:538 arg1 EMA 554:556 EMA 554:556 EMA 554:556 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 4 5 dep authorities 528:538 arg1 authorities 528:538 Health authorities 521:538 Health authorities (e.g. FDA and EMA) 521:557 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 4 5 dep authorities 528:538 arg1 FDA 546:548 FDA 546:548 FDA 546:548 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 9 6 theme special 1193:1199 arg1 focus 1201:1205 A special focus 1191:1205 A special focus 1191:1205 A special focus was given to N-glycosylation due to its potential to monitor the batch-to-batch consistency and alteration during the production bioprocess. 28371718 11 7 theme head-to-head 1561:1572 arg1 comparability 1574:1586 a head-to-head comparability 1559:1586 a head-to-head comparability of functional properties 1559:1611 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 6 8 theme middle-up 920:928 arg1 analysis 910:917 Intact mass analysis 898:917 Intact mass analysis 898:917 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 6 8 theme middle-up 920:928 arg1 approach 930:937 middle-up approach 920:937 middle-up approach 920:937 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 3 9 theme biosimilars 508:518 arg1 approval 490:497 the approval 486:497 the approval of these biosimilars 486:518 In correlation to their complex structure, an analytical challenge is facing the approval of these biosimilars. 28371718 2 10 theme similar 289:295 arg1 exits 335:339 similar or highly similar follow-on biologics exits 289:339 similar or highly similar follow-on biologics exits 289:339 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 11 11 theme properties 1602:1611 arg1 comparability 1574:1586 a head-to-head comparability 1559:1586 a head-to-head comparability of functional properties 1559:1611 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 9 12 theme production 1325:1334 arg1 bioprocess 1336:1345 the production bioprocess 1321:1345 the production bioprocess 1321:1345 A special focus was given to N-glycosylation due to its potential to monitor the batch-to-batch consistency and alteration during the production bioprocess. 28371718 10 13 theme profiles 1382:1389 arg1 product 1455:1461 the reference product 1441:1461 the reference product 1441:1461 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 10 13 theme profiles 1382:1389 arg1 Comparison 1348:1357 Comparison 1348:1357 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar 1348:1435 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 10 14 theme biosimilar 1426:1435 arg1 batches 1411:1417 three batches 1405:1417 three batches of the biosimilar 1405:1435 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 1 15 theme Immunglobolin 77:89 arg1 G 91:91 Immunglobolin G 77:91 Immunglobolin G (IgG)-based biopharmaceuticals 77:122 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 1 15 theme Immunglobolin 77:89 arg1 IgG 94:96 IgG 94:96 IgG 94:96 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 11 16 dep glycosylation 1656:1668 arg1 alteration 1670:1679 alteration 1670:1679 alteration 1670:1679 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 4 17 theme several 571:577 arg1 guidelines 579:588 several guidelines 571:588 several guidelines 571:588 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 6 18 from variants 1017:1024 arg1 biosimilar 1033:1042 the biosimilar 1029:1042 the biosimilar 1029:1042 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 5 19 theme Rituximab 860:868 arg1 PTMs 824:827 PTMs 824:827 PTMs 824:827 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 5 19 theme Rituximab 860:868 arg1 mass 784:787 intact mass 777:787 intact mass 777:787 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 5 19 theme Rituximab 860:868 arg1 modifications 809:821 post-translational modifications 790:821 post-translational modifications (PTMs) 790:828 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 5 19 theme Rituximab 860:868 arg1 structure 847:855 higher order structure 834:855 higher order structure of Rituximab 834:868 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 1 20 theme G 91:91 arg1 biopharmaceuticals 105:122 Immunglobolin G (IgG)-based biopharmaceuticals 77:122 Immunglobolin G (IgG)-based biopharmaceuticals 77:122 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 5 21 theme biosimilars 885:895 arg1 one 874:876 one 874:876 one 874:876 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 5 21 theme biosimilars 885:895 arg1 biosimilars 885:895 its biosimilars 881:895 its biosimilars 881:895 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 3 22 theme complex 433:439 arg1 structure 441:449 their complex structure 427:449 their complex structure 427:449 In correlation to their complex structure, an analytical challenge is facing the approval of these biosimilars. 28371718 6 23 theme lysine 1010:1015 arg1 variants 1017:1024 additional lysine variants 999:1024 additional lysine variants in the biosimilar 999:1042 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 6 24 theme mass 905:908 arg1 analysis 910:917 Intact mass analysis 898:917 Intact mass analysis 898:917 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 6 24 theme mass 905:908 arg1 approach 930:937 middle-up approach 920:937 middle-up approach 920:937 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 10 25 theme quantitative 1470:1481 arg1 variations 1483:1492 quantitative variations 1470:1492 quantitative variations 1470:1492 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 1 26 theme high 180:183 arg1 selectivity 192:202 their high target selectivity 174:202 their high target selectivity in different diseases 174:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 4 27 dep FDA 546:548 arg1 e.g. 541:544 e.g. 541:544 e.g. 541:544 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 2 28 theme follow-on 315:323 arg1 exits 335:339 similar or highly similar follow-on biologics exits 289:339 similar or highly similar follow-on biologics exits 289:339 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 1 29 theme target 185:190 arg1 selectivity 192:202 their high target selectivity 174:202 their high target selectivity in different diseases 174:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 5 30 theme higher 834:839 arg1 structure 847:855 higher order structure 834:855 higher order structure of Rituximab 834:868 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 6 31 theme additional 999:1008 arg1 variants 1017:1024 additional lysine variants 999:1024 additional lysine variants in the biosimilar 999:1042 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 12 32 theme acceptable 1829:1838 arg1 range 1840:1844 the acceptable range 1825:1844 the acceptable range for biosimilarity 1825:1862 The data affirm that the difference is still in the acceptable range for biosimilarity. 28371718 2 33 theme similar 307:313 arg1 exits 335:339 similar or highly similar follow-on biologics exits 289:339 similar or highly similar follow-on biologics exits 289:339 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 11 34 theme biosimilar 1712:1721 arg1 batches 1723:1729 the biosimilar batches 1708:1729 the biosimilar batches 1708:1729 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 2 35 theme biotherapeutics 373:387 arg1 patent 351:356 the patent 347:356 the patent of blockbuster biotherapeutics 347:387 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 0 36 theme Rituximab 23:31 arg1 originator 33:42 Rituximab originator 23:42 Rituximab originator 23:42 Comparability study of Rituximab originator and follow-on biopharmaceutical. 28371718 10 37 theme reference 1445:1453 arg1 product 1455:1461 the reference product 1441:1461 the reference product 1441:1461 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 1 38 theme -based 98:103 arg1 biopharmaceuticals 105:122 Immunglobolin G (IgG)-based biopharmaceuticals 77:122 Immunglobolin G (IgG)-based biopharmaceuticals 77:122 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 3 39 theme analytical 455:464 arg1 challenge 466:474 an analytical challenge 452:474 an analytical challenge 452:474 In correlation to their complex structure, an analytical challenge is facing the approval of these biosimilars. 28371718 11 40 theme glycosylation 1656:1668 arg1 impact 1646:1651 the impact 1642:1651 the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug 1642:1774 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 1 41 theme different 207:215 arg1 diseases 217:224 different diseases 207:224 different diseases 207:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 6 42 theme similar 976:982 arg1 glycoforms 984:993 similar glycoforms 976:993 similar glycoforms 976:993 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 5 43 theme state-of-the-art 727:742 arg1 analytics 744:752 state-of-the-art analytics 727:752 state-of-the-art analytics 727:752 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 4 44 theme process 649:655 arg1 changes 657:663 process changes 649:663 process changes 649:663 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 7 45 gly N-glycosylation 1049:1063 arg2 site 1065:1068 The N-glycosylation site 1045:1068 The N-glycosylation site 1045:1068 The N-glycosylation site was confirmed for both, the originator and the biosimilar. 28371718 5 46 theme order 841:845 arg1 structure 847:855 higher order structure 834:855 higher order structure of Rituximab 834:868 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 4 47 theme Health 521:526 arg1 EMA 554:556 EMA 554:556 EMA 554:556 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 4 47 theme Health 521:526 arg1 authorities 528:538 Health authorities 521:538 Health authorities (e.g. FDA and EMA) 521:557 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 4 47 theme Health 521:526 arg1 FDA 546:548 FDA 546:548 FDA 546:548 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 5 48 theme current 673:679 arg1 study 681:685 the current study 669:685 the current study 669:685 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 7 49 theme N-glycosylation 1049:1063 arg1 site 1065:1068 The N-glycosylation site 1045:1068 The N-glycosylation site 1045:1068 The N-glycosylation site was confirmed for both, the originator and the biosimilar. 28371718 0 50 theme follow-on 48:56 arg1 biopharmaceutical 58:74 follow-on biopharmaceutical 48:74 follow-on biopharmaceutical 48:74 Comparability study of Rituximab originator and follow-on biopharmaceutical. 28371718 6 51 theme subunit 950:956 arg1 analysis 958:965 subunit analysis 950:965 Intact mass analysis, middle-up approach as well as subunit analysis 898:965 Intact mass analysis, middle-up approach as well as subunit analysis revealed similar glycoforms but additional lysine variants in the biosimilar. 28371718 3 52 dep structure 441:449 arg1 correlation 412:422 correlation 412:422 correlation 412:422 In correlation to their complex structure, an analytical challenge is facing the approval of these biosimilars. 28371718 10 53 theme N-glycosylation 1366:1380 arg1 profiles 1382:1389 the N-glycosylation profiles 1362:1389 the N-glycosylation profiles obtained from three batches of the biosimilar 1362:1435 Comparison of the N-glycosylation profiles obtained from three batches of the biosimilar and the reference product showed quantitative variations, although the N-glycans were qualitatively similar. 28371718 8 54 theme order 1145:1149 arg1 structure 1151:1159 higher order structure 1138:1159 higher order structure 1138:1159 PTMs and higher order structure were confirmed to be similar. 28371718 1 55 theme pharmaceuticals 144:158 arg1 market 160:165 the pharmaceuticals market 140:165 the pharmaceuticals market due to their high target selectivity in different diseases 140:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 2 56 theme other 262:266 arg1 companies 268:276 other companies 262:276 other companies 262:276 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 8 57 theme higher 1138:1143 arg1 structure 1151:1159 higher order structure 1138:1159 higher order structure 1138:1159 PTMs and higher order structure were confirmed to be similar. 28371718 11 58 from impact 1646:1651 arg1 potency 1693:1699 potency 1693:1699 potency within the biosimilar batches and between originator and follow-on biodrug 1693:1774 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 5 59 theme physicochemical 688:702 arg1 characterization 704:719 physicochemical characterization 688:719 physicochemical characterization using state-of-the-art analytics 688:752 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 2 60 theme biologics 325:333 arg1 exits 335:339 similar or highly similar follow-on biologics exits 289:339 similar or highly similar follow-on biologics exits 289:339 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 5 61 theme intact 777:782 arg1 mass 784:787 intact mass 777:787 intact mass 777:787 In the current study, physicochemical characterization using state-of-the-art analytics was applied to analyse intact mass, post-translational modifications (PTMs) and higher order structure of Rituximab and one of its biosimilars. 28371718 1 62 from selectivity 192:202 arg1 diseases 217:224 different diseases 207:224 different diseases 207:224 Immunglobolin G (IgG)-based biopharmaceuticals are emerging on the pharmaceuticals market due to their high target selectivity in different diseases. 28371718 2 63 theme blockbuster 361:371 arg1 biotherapeutics 373:387 blockbuster biotherapeutics 361:387 blockbuster biotherapeutics 361:387 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 9 64 theme batch-to-batch 1272:1285 arg1 consistency 1287:1297 the batch-to-batch consistency 1268:1297 the batch-to-batch consistency 1268:1297 A special focus was given to N-glycosylation due to its potential to monitor the batch-to-batch consistency and alteration during the production bioprocess. 28371718 4 65 theme quality 609:615 arg1 attributes 617:626 critical quality attributes 600:626 critical quality attributes 600:626 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28371718 11 66 theme PTMs 1685:1688 arg1 impact 1646:1651 the impact 1642:1651 the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug 1642:1774 Furthermore, a head-to-head comparability of functional properties was performed to investigate the impact of glycosylation alteration and PTMs on potency within the biosimilar batches and between originator and follow-on biodrug. 28371718 2 67 theme growing 242:248 arg1 interest 250:257 a growing interest 240:257 a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics 240:387 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 2 67 theme growing 242:248 arg1 expire 401:406 expire 401:406 to expire 398:406 In parallel, a growing interest by other companies to produce similar or highly similar follow-on biologics exits, once the patent of blockbuster biotherapeutics is about to expire. 28371718 4 68 theme critical 600:607 arg1 attributes 617:626 critical quality attributes 600:626 critical quality attributes 600:626 Health authorities (e.g. FDA and EMA) have issued several guidelines to define critical quality attributes during manufacturing process changes. 28938118 2 0 theme different 212:220 arg1 organisms 222:230 different organisms 212:230 different organisms that install these complex and highly variable structures 212:288 This SnapShot presents the core pathways from different organisms that install these complex and highly variable structures. 28938118 1 1 with modification 82:93 arg1 carbohydrates 112:124 carbohydrates 112:124 carbohydrates 112:124 Post-translational modification of proteins with carbohydrates shapes their localization and function. 28938118 1 2 theme proteins 98:105 arg1 modification 82:93 Post-translational modification 63:93 Post-translational modification of proteins with carbohydrates 63:124 Post-translational modification of proteins with carbohydrates shapes their localization and function. 28938118 2 3 theme variable 270:277 arg1 structures 279:288 these complex and highly variable structures 245:288 these complex and highly variable structures 245:288 This SnapShot presents the core pathways from different organisms that install these complex and highly variable structures. 28938118 2 4 theme complex 251:257 arg1 structures 279:288 these complex and highly variable structures 245:288 these complex and highly variable structures 245:288 This SnapShot presents the core pathways from different organisms that install these complex and highly variable structures. 28938118 1 5 theme Post-translational 63:80 arg1 modification 82:93 Post-translational modification 63:93 Post-translational modification of proteins with carbohydrates 63:124 Post-translational modification of proteins with carbohydrates shapes their localization and function. 28938118 2 6 from organisms 222:230 arg1 pathways 198:205 the core pathways 189:205 the core pathways from different organisms that install these complex and highly variable structures 189:288 This SnapShot presents the core pathways from different organisms that install these complex and highly variable structures. 28938118 2 7 theme core 193:196 arg1 pathways 198:205 the core pathways 189:205 the core pathways from different organisms that install these complex and highly variable structures 189:288 This SnapShot presents the core pathways from different organisms that install these complex and highly variable structures. 28938118 0 8 theme Processing 26:35 arg1 Pathways 37:44 N-Glycosylation Processing Pathways 10:44 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 0:61 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 28938118 0 9 dep SnapShot 0:7 arg1 Pathways 37:44 N-Glycosylation Processing Pathways 10:44 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 0:61 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 28938118 0 10 theme N-Glycosylation 10:24 arg1 Pathways 37:44 N-Glycosylation Processing Pathways 10:44 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 0:61 SnapShot: N-Glycosylation Processing Pathways across Kingdoms. 25462875 0 0 theme insect 96:101 arg1 cells 103:107 insect cells 96:107 insect cells 96:107 Engineering β1,4-galactosyltransferase I to reduce secretion and enhance N-glycan elongation in insect cells. 25462875 1 1 gly glycoprotein 191:202 arg1 glycoprotein 191:202 glycoprotein glycans 191:210 glycoprotein glycans 191:210 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 4 2 theme secreted 702:709 arg1 levels 722:727 higher levels 715:727 higher levels of intracellular B4GALT1 activity 715:761 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 4 2 theme secreted 702:709 arg1 levels 692:697 lower levels 686:697 lower levels of secreted 686:709 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 5 3 theme other 879:883 arg1 sequences 900:908 all other animal B4GALT1 sequences 875:908 all other animal B4GALT1 sequences 875:908 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 1 4 theme proteolytic 270:280 arg1 cleavage 282:289 proteolytic cleavage 270:289 proteolytic cleavage in its stem domain 270:308 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 7 5 theme structures 1317:1326 arg1 N-glycans 1261:1269 terminally galactosylated N-glycans 1235:1269 terminally galactosylated N-glycans 1235:1269 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 5 theme structures 1317:1326 arg1 levels 1190:1195 the levels 1186:1195 the levels of intracellular B4GALT1 activity 1186:1229 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 5 theme structures 1317:1326 arg1 appearance 1296:1305 the appearance 1292:1305 the appearance of hybrid structures 1292:1326 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 5 theme structures 1317:1326 arg1 fucosylation 1346:1357 reduced core fucosylation 1333:1357 reduced core fucosylation 1333:1357 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 5 theme structures 1317:1326 arg1 branching 1281:1289 N-glycan branching 1272:1289 N-glycan branching 1272:1289 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 4 6 theme native 772:777 arg1 enzyme 779:784 the native enzyme 768:784 the native enzyme 768:784 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 6 7 from impact 987:992 arg1 processing 1111:1120 N-glycan processing 1102:1120 N-glycan processing in insect cells 1102:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 6 7 from impact 987:992 arg1 levels 1091:1096 intracellular B4GALT1 activity levels 1060:1096 intracellular B4GALT1 activity levels 1060:1096 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 7 8 theme B4GALT1 1214:1220 arg1 activity 1222:1229 intracellular B4GALT1 activity 1200:1229 intracellular B4GALT1 activity 1200:1229 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 3 9 theme human 555:559 arg1 α1,3-fucosyltransferase 561:583 human α1,3-fucosyltransferase 7 555:585 human α1,3-fucosyltransferase 7 (FUT7) 555:592 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 3 9 theme human 555:559 arg1 FUT7 588:591 FUT7 588:591 FUT7 588:591 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 3 10 theme cytoplasmic/transmembrane/stem 483:512 arg1 domains 520:526 the cytoplasmic/transmembrane/stem (CTS) domains 479:526 the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 479:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 3 10 theme cytoplasmic/transmembrane/stem 483:512 arg1 B4GALT1 531:537 B4GALT1 531:537 B4GALT1 531:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 5 11 theme animal 885:890 arg1 sequences 900:908 all other animal B4GALT1 sequences 875:908 all other animal B4GALT1 sequences 875:908 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 4 12 theme insect 664:669 arg1 cells 671:675 insect cells 664:675 insect cells 664:675 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 6 13 theme N-glycan 1102:1109 arg1 processing 1111:1120 N-glycan processing 1102:1120 N-glycan processing in insect cells 1102:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 6 14 theme insect 1125:1130 arg1 cells 1132:1136 insect cells 1125:1136 insect cells 1125:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 1 15 theme glycoprotein 191:202 arg1 glycans 204:210 glycoprotein glycans 191:210 glycoprotein glycans 191:210 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 7 16 theme N-glycan 1272:1279 arg1 branching 1281:1289 N-glycan branching 1272:1289 N-glycan branching 1272:1289 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 5 17 contain had 836:838 arg1 B4GALT1 810:816 the B4GALT1 806:816 the B4GALT1 used in our study 806:834 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 5 17 contain had 836:838 arg2 leucine 842:848 a leucine 840:848 a leucine 840:848 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 4 18 theme B4GALT1 746:752 arg1 activity 754:761 intracellular B4GALT1 activity 732:761 intracellular B4GALT1 activity 732:761 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 7 19 theme galactosylated 1246:1259 arg1 N-glycans 1261:1269 terminally galactosylated N-glycans 1235:1269 terminally galactosylated N-glycans 1235:1269 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 8 20 from elongation 1475:1484 arg1 cells 1496:1500 insect cells 1489:1500 insect cells 1489:1500 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 2 21 dep B4GALT1 344:350 arg1 block 355:359 block 355:359 to block cleavage and secretion 352:382 We hypothesized that engineering B4GALT1 to block cleavage and secretion would enhance its retention and, therefore, its function. 25462875 1 22 theme stem 298:301 arg1 domain 303:308 its stem domain 294:308 its stem domain 294:308 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 1 23 from cleavage 282:289 arg1 domain 303:308 its stem domain 294:308 its stem domain 294:308 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 1 24 theme β1,4-galactosyltransferase 110:135 arg1 enzyme 169:174 a Golgi-resident enzyme 152:174 a Golgi-resident enzyme that elongates glycoprotein glycans 152:210 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 1 24 theme β1,4-galactosyltransferase 110:135 arg1 B4GALT1 140:146 B4GALT1 140:146 B4GALT1 140:146 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 1 24 theme β1,4-galactosyltransferase 110:135 arg1 I 137:137 β1,4-galactosyltransferase I 110:137 β1,4-galactosyltransferase I (B4GALT1) 110:147 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 4 25 theme intracellular 732:744 arg1 activity 754:761 intracellular B4GALT1 activity 732:761 intracellular B4GALT1 activity 732:761 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 8 26 theme insect 1489:1494 arg1 cells 1496:1500 insect cells 1489:1500 insect cells 1489:1500 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 5 27 theme aromatic 918:925 arg1 acid 933:936 an aromatic amino acid 915:936 an aromatic amino acid 915:936 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 4 28 theme activity 754:761 arg1 levels 722:727 higher levels 715:727 higher levels of intracellular B4GALT1 activity 715:761 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 4 28 theme activity 754:761 arg1 levels 692:697 lower levels 686:697 lower levels of secreted 686:709 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 4 29 from Expression 630:639 arg1 cells 671:675 insect cells 664:675 insect cells 664:675 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 6 30 theme CTS 1010:1012 arg1 domains 1014:1020 the CTS domains 1006:1020 the CTS domains 1006:1020 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 5 31 theme B4GALT1 892:898 arg1 sequences 900:908 all other animal B4GALT1 sequences 875:908 all other animal B4GALT1 sequences 875:908 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 6 32 theme activity 1082:1089 arg1 levels 1091:1096 intracellular B4GALT1 activity levels 1060:1096 intracellular B4GALT1 activity levels 1060:1096 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 6 33 from processing 1111:1120 arg1 cells 1132:1136 insect cells 1125:1136 insect cells 1125:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 4 34 theme FUT7-CTS-B4GALT1 644:659 arg1 Expression 630:639 Expression 630:639 Expression of FUT7-CTS-B4GALT1 in insect cells 630:675 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 7 35 theme hybrid 1310:1315 arg1 structures 1317:1326 hybrid structures 1310:1326 hybrid structures 1310:1326 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 8 36 used used 1450:1453 arg2 B4GALT1 1378:1384 engineering B4GALT1 1366:1384 engineering B4GALT1 to reduce its cleavage and secretion 1366:1421 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 8 36 used used 1450:1453 arg2 approach 1429:1436 an approach 1426:1436 an approach that can be used to enhance N-glycan elongation in insect cells 1426:1500 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 0 37 from elongation 82:91 arg1 cells 103:107 insect cells 96:107 insect cells 96:107 Engineering β1,4-galactosyltransferase I to reduce secretion and enhance N-glycan elongation in insect cells. 25462875 8 38 theme engineering 1366:1376 arg1 approach 1429:1436 an approach 1426:1436 an approach that can be used to enhance N-glycan elongation in insect cells 1426:1500 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 8 38 theme engineering 1366:1376 arg1 B4GALT1 1378:1384 engineering B4GALT1 1366:1384 engineering B4GALT1 to reduce its cleavage and secretion 1366:1421 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 5 39 contain have 910:913 arg2 acid 933:936 an aromatic amino acid 915:936 an aromatic amino acid 915:936 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 5 39 contain have 910:913 arg1 sequences 900:908 all other animal B4GALT1 sequences 875:908 all other animal B4GALT1 sequences 875:908 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 4 40 theme higher 715:720 arg1 levels 722:727 higher levels 715:727 higher levels of intracellular B4GALT1 activity 715:761 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 7 41 gly fucosylation 1346:1357 arg1 activity 1222:1229 intracellular B4GALT1 activity 1200:1229 intracellular B4GALT1 activity 1200:1229 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 41 gly fucosylation 1346:1357 arg1 structures 1317:1326 hybrid structures 1310:1326 hybrid structures 1310:1326 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 1 42 theme Golgi-resident 154:167 arg1 enzyme 169:174 a Golgi-resident enzyme 152:174 a Golgi-resident enzyme that elongates glycoprotein glycans 152:210 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 1 42 theme Golgi-resident 154:167 arg1 I 137:137 β1,4-galactosyltransferase I 110:137 β1,4-galactosyltransferase I (B4GALT1) 110:147 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 6 43 from levels 1091:1096 arg1 cells 1132:1136 insect cells 1125:1136 insect cells 1125:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 3 44 theme CTS 515:517 arg1 domains 520:526 the cytoplasmic/transmembrane/stem (CTS) domains 479:526 the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 479:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 3 44 theme CTS 515:517 arg1 B4GALT1 531:537 B4GALT1 531:537 B4GALT1 531:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 7 45 theme reduced 1333:1339 arg1 fucosylation 1346:1357 reduced core fucosylation 1333:1357 reduced core fucosylation 1333:1357 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 6 46 theme B4GALT1 1074:1080 arg1 levels 1091:1096 intracellular B4GALT1 activity levels 1060:1096 intracellular B4GALT1 activity levels 1060:1096 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 5 47 theme amino 927:931 arg1 acid 933:936 an aromatic amino acid 915:936 an aromatic amino acid 915:936 We also noted that the B4GALT1 used in our study had a leucine at position 282, whereas all other animal B4GALT1 sequences have an aromatic amino acid at this position. 25462875 8 48 dep B4GALT1 1378:1384 arg1 reduce 1389:1394 reduce 1389:1394 to reduce its cleavage and secretion 1386:1421 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 7 49 theme core 1341:1344 arg1 fucosylation 1346:1357 reduced core fucosylation 1333:1357 reduced core fucosylation 1333:1357 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 3 50 theme B4GALT1 531:537 arg1 domains 520:526 the cytoplasmic/transmembrane/stem (CTS) domains 479:526 the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 479:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 3 50 theme B4GALT1 531:537 arg1 B4GALT1 531:537 B4GALT1 531:537 B4GALT1 531:537 To test this hypothesis, we replaced the cytoplasmic/transmembrane/stem (CTS) domains of B4GALT1 with those from human α1,3-fucosyltransferase 7 (FUT7), which is not cleaved and secreted. 25462875 1 51 theme enzyme 241:246 arg1 subpopulation 219:231 a subpopulation 217:231 a subpopulation of this enzyme 217:246 β1,4-galactosyltransferase I (B4GALT1) is a Golgi-resident enzyme that elongates glycoprotein glycans, but a subpopulation of this enzyme is secreted following proteolytic cleavage in its stem domain. 25462875 7 52 theme activity 1222:1229 arg1 N-glycans 1261:1269 terminally galactosylated N-glycans 1235:1269 terminally galactosylated N-glycans 1235:1269 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 52 theme activity 1222:1229 arg1 levels 1190:1195 the levels 1186:1195 the levels of intracellular B4GALT1 activity 1186:1229 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 52 theme activity 1222:1229 arg1 appearance 1296:1305 the appearance 1292:1305 the appearance of hybrid structures 1292:1326 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 52 theme activity 1222:1229 arg1 fucosylation 1346:1357 reduced core fucosylation 1333:1357 reduced core fucosylation 1333:1357 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 7 52 theme activity 1222:1229 arg1 branching 1281:1289 N-glycan branching 1272:1289 N-glycan branching 1272:1289 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 25462875 6 53 theme amino 1030:1034 arg1 acid 1036:1039 the amino acid 1026:1039 the amino acid 1026:1039 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 6 54 theme combined 978:985 arg1 impact 987:992 the combined impact 974:992 the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells 974:1136 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 0 55 theme N-glycan 73:80 arg1 elongation 82:91 N-glycan elongation 73:91 N-glycan elongation in insect cells 73:107 Engineering β1,4-galactosyltransferase I to reduce secretion and enhance N-glycan elongation in insect cells. 25462875 8 56 theme N-glycan 1466:1473 arg1 elongation 1475:1484 N-glycan elongation 1466:1484 N-glycan elongation in insect cells 1466:1500 Thus, engineering B4GALT1 to reduce its cleavage and secretion is an approach that can be used to enhance N-glycan elongation in insect cells. 25462875 6 57 theme intracellular 1060:1072 arg1 levels 1091:1096 intracellular B4GALT1 activity levels 1060:1096 intracellular B4GALT1 activity levels 1060:1096 Thus, we examined the combined impact of changing the CTS domains and the amino acid at position 282 on intracellular B4GALT1 activity levels and N-glycan processing in insect cells. 25462875 4 58 theme lower 686:690 arg1 levels 692:697 lower levels 686:697 lower levels of secreted 686:709 Expression of FUT7-CTS-B4GALT1 in insect cells produced lower levels of secreted and higher levels of intracellular B4GALT1 activity than the native enzyme. 25462875 7 59 theme intracellular 1200:1212 arg1 activity 1222:1229 intracellular B4GALT1 activity 1200:1229 intracellular B4GALT1 activity 1200:1229 The results demonstrated a correlation between the levels of intracellular B4GALT1 activity and terminally galactosylated N-glycans, N-glycan branching, the appearance of hybrid structures, and reduced core fucosylation. 28299519 4 0 theme molecular 502:510 arg1 simulations 521:531 molecular docking and molecular dynamics simulations 480:531 simulations 521:531 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 1 1 theme Crataeva 152:159 arg1 bark 167:170 Crataeva tapia bark 152:170 Crataeva tapia bark 152:170 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 9 2 theme better 1392:1397 arg1 comprehension 1399:1411 a better comprehension 1390:1411 a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures 1390:1501 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 6 3 theme monosaccharides 1062:1076 arg1 states 1048:1053 α- and β-anomeric states 1030:1053 α- and β-anomeric states of the monosaccharides 1030:1076 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 5 4 theme CrataBL 811:817 arg1 moiety 827:832 CrataBL protein moiety 811:832 CrataBL protein moiety 811:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 5 5 theme protein 819:825 arg1 moiety 827:832 CrataBL protein moiety 811:832 CrataBL protein moiety 811:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 7 6 theme binding 1218:1224 arg1 preference 1226:1235 the binding preference 1214:1235 the binding preference for glucose over galactose 1214:1262 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 1 7 theme tapia 161:165 arg1 bark 167:170 Crataeva tapia bark 152:170 Crataeva tapia bark 152:170 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 0 8 theme lectin 96:101 arg1 CrataBL 103:109 the plant lectin CrataBL 86:109 the plant lectin CrataBL 86:109 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 4 9 theme binding 675:681 arg1 specificity 683:693 its binding specificity 671:693 its binding specificity 671:693 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 0 10 theme plant 90:94 arg1 CrataBL 103:109 the plant lectin CrataBL 86:109 the plant lectin CrataBL 86:109 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 4 11 theme molecular 480:488 arg1 docking 490:496 molecular docking and molecular dynamics simulations 480:531 docking 490:496 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 9 12 theme lectins 1420:1426 arg1 comprehension 1399:1411 a better comprehension 1390:1411 a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures 1390:1501 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 6 13 theme lectin 849:854 arg1 activity 856:863 its lectin activity 845:863 its lectin activity 845:863 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 9 14 theme site 1487:1490 arg1 structures 1492:1501 carbohydrate-binding site structures 1466:1501 carbohydrate-binding site structures 1466:1501 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 4 15 theme aqueous 617:623 arg1 solutions 625:633 aqueous solutions 617:633 aqueous solutions 617:633 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 1 16 attach isolated 138:145 arg2 CrataBL 112:118 CrataBL 112:118 CrataBL 112:118 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 16 attach isolated 138:145 arg1 bark 167:170 Crataeva tapia bark 152:170 Crataeva tapia bark 152:170 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 16 attach isolated 138:145 arg2 glycoprotein 125:136 a glycoprotein 123:136 a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites 123:208 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 3 17 gly glycosylation 370:382 arg2 sites 418:422 glycosylation or CrataBL monosaccharide-binding sites 370:422 sites 418:422 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 3 18 theme specificity 443:453 arg1 effects 359:365 the effects 355:365 the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity 355:453 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 0 19 theme CrataBL 103:109 arg1 Insights 0:7 Insights 0:7 Insights into the effects of glycosylation 0:41 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 0 19 theme CrataBL 103:109 arg1 activity 74:81 the monosaccharide-binding activity 47:81 the monosaccharide-binding activity of the plant lectin CrataBL 47:109 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 1 20 gly N-glycosylation 188:202 arg2 sites 204:208 two N-glycosylation sites 184:208 two N-glycosylation sites 184:208 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 20 gly N-glycosylation 188:202 arg2 two 184:186 two 184:186 two 184:186 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 4 21 from conformation 588:599 arg1 solutions 625:633 aqueous solutions 617:633 aqueous solutions 617:633 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 6 22 theme structure-based 978:992 arg1 information 1011:1021 sequence-based, structure-based and evolutionary information 962:1021 sequence-based, structure-based and evolutionary information 962:1021 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 6 23 theme binding 928:934 arg1 sites 936:940 two possible binding sites 915:940 two possible binding sites 915:940 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 6 24 theme β-anomeric 1037:1046 arg1 states 1048:1053 α- and β-anomeric states 1030:1053 α- and β-anomeric states of the monosaccharides 1030:1076 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 4 25 dep conformation 588:599 arg1 the 563:565 the 563:565 the 563:565 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 3 26 theme glycosylation 370:382 arg1 sites 418:422 glycosylation or CrataBL monosaccharide-binding sites 370:422 sites 418:422 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 4 27 theme CrataBL 580:586 arg1 conformation 588:599 glycosylated CrataBL conformation 567:599 glycosylated CrataBL conformation 567:599 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 9 28 theme structures 1492:1501 arg1 comprehension 1399:1411 a better comprehension 1390:1411 a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures 1390:1501 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 7 29 theme positively-charged 1175:1192 arg1 acids 1200:1204 positively-charged amino acids 1175:1204 positively-charged amino acids 1175:1204 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 3 30 theme monosaccharide 428:441 arg1 specificity 443:453 monosaccharide specificity 428:453 monosaccharide specificity 428:453 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 4 31 theme glycosylated 567:578 arg1 conformation 588:599 glycosylated CrataBL conformation 567:599 glycosylated CrataBL conformation 567:599 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 5 32 link N-linked 789:796 arg1 glycans 798:804 N-linked glycans 789:804 N-linked glycans 789:804 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 1 33 theme N-glycosylation 188:202 arg1 sites 204:208 two N-glycosylation sites 184:208 two N-glycosylation sites 184:208 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 6 34 theme α- 1030:1031 arg1 states 1048:1053 α- and β-anomeric states 1030:1053 α- and β-anomeric states of the monosaccharides 1030:1076 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 5 35 theme local 731:735 arg1 effects 778:784 both local and distant conformational stabilization effects 726:784 both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety 726:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 6 36 theme possible 919:926 arg1 sites 936:940 two possible binding sites 915:940 two possible binding sites 915:940 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 3 37 theme monosaccharide-binding 395:416 arg1 sites 418:422 glycosylation or CrataBL monosaccharide-binding sites 370:422 sites 418:422 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 1 38 gly glycoprotein 125:136 arg1 CrataBL 112:118 CrataBL 112:118 CrataBL 112:118 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 38 gly glycoprotein 125:136 arg1 glycoprotein 125:136 a glycoprotein 123:136 a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites 123:208 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 39 contain containing 173:182 arg1 bark 167:170 Crataeva tapia bark 152:170 Crataeva tapia bark 152:170 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 1 39 contain containing 173:182 arg2 sites 204:208 two N-glycosylation sites 184:208 two N-glycosylation sites 184:208 CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. 28299519 3 40 theme sites 418:422 arg1 effects 359:365 the effects 355:365 the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity 355:453 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 9 41 theme carbohydrate-binding 1466:1485 arg1 structures 1492:1501 carbohydrate-binding site structures 1466:1501 carbohydrate-binding site structures 1466:1501 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 5 42 theme distant 741:747 arg1 effects 778:784 both local and distant conformational stabilization effects 726:784 both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety 726:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 3 43 from information 340:350 arg1 effects 359:365 the effects 355:365 the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity 355:453 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 9 44 theme monosaccharide-binding 1428:1449 arg1 activities 1451:1460 monosaccharide-binding activities 1428:1460 monosaccharide-binding activities 1428:1460 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 4 45 gly glycosylated 567:578 arg1 conformation 588:599 glycosylated CrataBL conformation 567:599 glycosylated CrataBL conformation 567:599 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 4 46 from dynamics 605:612 arg1 solutions 625:633 aqueous solutions 617:633 aqueous solutions 617:633 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 5 47 theme conformational 749:762 arg1 effects 778:784 both local and distant conformational stabilization effects 726:784 both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety 726:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 5 48 theme glycans 798:804 arg1 effects 778:784 both local and distant conformational stabilization effects 726:784 both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety 726:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 7 49 theme dynamics 1137:1144 arg1 simulations 1146:1156 molecular dynamics simulations 1127:1156 molecular dynamics simulations 1127:1156 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 3 50 theme CrataBL 387:393 arg1 sites 418:422 glycosylation or CrataBL monosaccharide-binding sites 370:422 sites 418:422 However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. 28299519 6 51 theme evolutionary 998:1009 arg1 information 1011:1021 sequence-based, structure-based and evolutionary information 962:1021 sequence-based, structure-based and evolutionary information 962:1021 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 5 52 theme obtained 700:707 arg1 results 709:715 The obtained results 696:715 The obtained results 696:715 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 6 53 theme sequence-based 962:975 arg1 information 1011:1021 sequence-based, structure-based and evolutionary information 962:1021 sequence-based, structure-based and evolutionary information 962:1021 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 7 54 theme amino 1194:1198 arg1 acids 1200:1204 positively-charged amino acids 1175:1204 positively-charged amino acids 1175:1204 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 8 55 theme possible 1294:1301 arg1 preference 1303:1312 a possible preference 1292:1312 a possible preference for β-monosaccharides 1292:1334 In addition, a possible preference for β-monosaccharides was proposed. 28299519 9 56 dep lectins 1420:1426 arg1 activities 1451:1460 monosaccharide-binding activities 1428:1460 monosaccharide-binding activities 1428:1460 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 0 57 theme glycosylation 29:41 arg1 effects 18:24 the effects 14:24 the effects of glycosylation 14:41 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 6 58 theme docking 876:882 arg1 calculations 884:895 molecular docking calculations 866:895 molecular docking calculations 866:895 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 7 59 theme obtained 1083:1090 arg1 poses 1092:1096 The obtained poses 1079:1096 The obtained poses 1079:1096 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 9 60 theme Such 1350:1353 arg1 data 1355:1358 Such data 1350:1358 Such data 1350:1358 Such data are expected to contribute to a better comprehension of the lectins monosaccharide-binding activities and carbohydrate-binding site structures. 28299519 5 61 theme N-linked 789:796 arg1 glycans 798:804 N-linked glycans 789:804 N-linked glycans 789:804 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 28299519 6 62 theme molecular 866:874 arg1 calculations 884:895 molecular docking calculations 866:895 molecular docking calculations 866:895 Regarding its lectin activity, molecular docking calculations were performed in two possible binding sites, identified through sequence-based, structure-based and evolutionary information, using α- and β-anomeric states of the monosaccharides. 28299519 2 63 theme lectin 245:250 arg1 activity 252:259 lectin activity 245:259 lectin activity 245:259 It has been identified to present lectin activity with some specificity for binding glucose over galactose. 28299519 4 64 theme molecular 651:659 arg1 basis 661:665 the molecular basis 647:665 the molecular basis for its binding specificity 647:693 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 0 65 theme monosaccharide-binding 51:72 arg1 activity 74:81 the monosaccharide-binding activity 47:81 the monosaccharide-binding activity of the plant lectin CrataBL 47:109 Insights into the effects of glycosylation and the monosaccharide-binding activity of the plant lectin CrataBL. 28299519 4 66 theme dynamics 512:519 arg1 simulations 521:531 molecular docking and molecular dynamics simulations 480:531 simulations 521:531 Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity. 28299519 7 67 theme molecular 1127:1135 arg1 simulations 1146:1156 molecular dynamics simulations 1127:1156 molecular dynamics simulations 1127:1156 The obtained poses were further refined through molecular dynamics simulations, suggesting that positively-charged amino acids dictate the binding preference for glucose over galactose in both sites. 28299519 5 68 theme stabilization 764:776 arg1 effects 778:784 both local and distant conformational stabilization effects 726:784 both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety 726:832 The obtained results indicate both local and distant conformational stabilization effects of N-linked glycans over CrataBL protein moiety. 26154505 4 0 theme structures 837:846 arg1 profiling 811:819 detailed profiling 802:819 detailed profiling of the N-glycan structures using mass spectra (MS) analysis 802:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 4 1 theme mass 854:857 arg1 analysis 872:879 mass spectra (MS) analysis 854:879 mass spectra (MS) analysis 854:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 6 2 theme N-glycans 1245:1253 arg1 % 1230:1230 approximately 92% 1214:1230 approximately 92% of the total N-glycans on the resulting EPO 1214:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 6 2 theme N-glycans 1245:1253 arg1 N-glycans 1245:1253 the total N-glycans 1235:1253 the total N-glycans on the resulting EPO 1235:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 8 3 theme N-glycan 1675:1682 arg1 complexity 1694:1703 N-glycan branching complexity 1675:1703 N-glycan branching complexity 1675:1703 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 3 4 theme glycans 709:715 arg1 content 692:698 the sialic acid content 676:698 the sialic acid content of total glycans on EPO 676:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 8 5 theme complex 1761:1767 arg1 structures 1778:1787 tailored complex N-glycan structures 1752:1787 tailored complex N-glycan structures on therapeutic glycoproteins in the future 1752:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 7 6 theme parental 1501:1508 arg1 CHO-K1 1510:1515 the wild-type parental CHO-K1 1487:1515 the wild-type parental CHO-K1 1487:1515 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 2 7 from impact 349:354 arg1 N-glycosylation 397:411 the N-glycosylation 393:411 the N-glycosylation of recombinant erythropoietin (rEPO) 393:448 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 7 8 theme sugar 1454:1458 arg1 chains 1460:1465 total sugar chains 1448:1465 total sugar chains 1448:1465 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 8 9 theme therapeutic 1792:1802 arg1 glycoproteins 1804:1816 therapeutic glycoproteins 1792:1816 therapeutic glycoproteins in the future 1792:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 6 10 theme MS 1294:1295 arg1 analysis 1297:1304 MS analysis 1294:1304 MS analysis 1294:1304 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 3 11 from content 692:698 arg1 EPO 720:722 EPO 720:722 EPO 720:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 3 12 theme CHO 585:587 arg1 proteins 598:605 CHO cellular proteins 585:605 CHO cellular proteins 585:605 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 8 13 gly glycoproteins 1804:1816 arg1 glycoproteins 1804:1816 therapeutic glycoproteins 1792:1816 therapeutic glycoproteins in the future 1792:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 5 14 dep UDP-N-acetylglucosamine 925:947 arg1 GnTIV/Mgat4 1006:1016 GnTIV/Mgat4 1006:1016 GnTIV/Mgat4 1006:1016 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 14 dep UDP-N-acetylglucosamine 925:947 arg1 β1,4-N-acetylglucosaminyltransferase 968:1003 α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase 950:1003 α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) 950:1017 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 14 dep UDP-N-acetylglucosamine 925:947 arg1 GnTV/Mgat5 1103:1112 GnTV/Mgat5 1103:1112 GnTV/Mgat5 1103:1112 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 14 dep UDP-N-acetylglucosamine 925:947 arg1 UDP-N-acetylglucosamine 1023:1045 UDP-N-acetylglucosamine 1023:1045 UDP-N-acetylglucosamine 1023:1045 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 14 dep UDP-N-acetylglucosamine 925:947 arg1 β1,6-N-acetylglucosaminyltransferase 1065:1100 α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase 1047:1100 α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5) 1047:1113 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 3 15 theme SNA 622:624 arg1 analysis 633:640 SNA lectin analysis 622:640 SNA lectin analysis 622:640 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 2 16 theme Chinese 509:515 arg1 cells 540:544 Chinese hamster ovary (CHO-K1) cells 509:544 Chinese hamster ovary (CHO-K1) cells 509:544 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 0 17 theme erythropoietin 61:74 arg1 branching 85:93 enhanced erythropoietin N-glycan branching 52:93 enhanced erythropoietin N-glycan branching 52:93 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 18 theme complex 147:153 arg1 N-glycans 155:163 complex N-glycans 147:163 complex N-glycans 147:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 4 19 from increase 746:753 arg1 content 770:776 sialic acid content 758:776 sialic acid content 758:776 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 2 20 theme CHO-K1 532:537 arg1 cells 540:544 Chinese hamster ovary (CHO-K1) cells 509:544 Chinese hamster ovary (CHO-K1) cells 509:544 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 8 21 theme coordinated 1531:1541 arg1 overexpression 1543:1556 coordinated overexpression 1531:1556 coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines 1531:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 2 22 theme ovary 525:529 arg1 cells 540:544 Chinese hamster ovary (CHO-K1) cells 509:544 Chinese hamster ovary (CHO-K1) cells 509:544 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 2 23 gly N-glycosylation 397:411 arg1 erythropoietin 428:441 recombinant erythropoietin 416:441 recombinant erythropoietin (rEPO) 416:448 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 2 23 gly N-glycosylation 397:411 arg1 rEPO 444:447 rEPO 444:447 rEPO 444:447 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 2 24 theme human 453:457 arg1 ST6Gal1 483:489 ST6Gal1 483:489 ST6Gal1 483:489 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 2 24 theme human 453:457 arg1 α2,6-sialyltransferase 459:480 a human α2,6-sialyltransferase 451:480 a human α2,6-sialyltransferase (ST6Gal1) 451:490 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 0 25 theme Chinese 20:26 arg1 cells 42:46 Chinese hamster ovary cells 20:46 Chinese hamster ovary cells 20:46 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 26 theme glycoproteins 286:298 arg1 activity 237:244 biological activity 226:244 biological activity 226:244 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 1 26 theme glycoproteins 286:298 arg1 lifetime 262:269 circulatory lifetime 250:269 circulatory lifetime 250:269 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 7 27 theme engineered 1359:1368 arg1 cells 1370:1374 these engineered cells 1353:1374 these engineered cells 1353:1374 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 8 28 theme first 1602:1606 arg1 time 1608:1611 the first time 1598:1611 the first time in model CHO-K1 cell lines 1598:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 8 29 from structures 1778:1787 arg1 glycoproteins 1804:1816 therapeutic glycoproteins 1792:1816 therapeutic glycoproteins in the future 1792:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 2 30 theme erythropoietin 428:441 arg1 N-glycosylation 397:411 the N-glycosylation 393:411 the N-glycosylation of recombinant erythropoietin (rEPO) 393:448 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 0 31 theme ovary 36:40 arg1 cells 42:46 Chinese hamster ovary cells 20:46 Chinese hamster ovary cells 20:46 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 32 theme therapeutic 303:313 arg1 interest 315:322 therapeutic interest 303:322 therapeutic interest 303:322 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 6 33 theme tetraantennary 1177:1190 arg1 N-glycans 1192:1200 Tri- and tetraantennary N-glycans 1168:1200 Tri- and tetraantennary N-glycans 1168:1200 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 8 34 theme CHO-K1 1622:1627 arg1 lines 1634:1638 model CHO-K1 cell lines 1616:1638 model CHO-K1 cell lines 1616:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 4 35 theme sialic 758:763 arg1 content 770:776 sialic acid content 758:776 sialic acid content 758:776 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 1 36 from acid 119:122 arg1 N-glycans 155:163 complex N-glycans 147:163 complex N-glycans 147:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 7 37 from cells 1370:1374 arg1 content 1332:1338 sialic acid content 1320:1338 sialic acid content of rEPO from these engineered cells 1320:1374 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 6 38 theme Tri- 1168:1171 arg1 N-glycans 1192:1200 Tri- and tetraantennary N-glycans 1168:1200 Tri- and tetraantennary N-glycans 1168:1200 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 2 39 theme expression 379:388 arg1 impact 349:354 the impact 345:354 the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO) 345:448 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 8 40 from time 1608:1611 arg1 lines 1634:1638 model CHO-K1 cell lines 1616:1638 model CHO-K1 cell lines 1616:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 4 41 theme N-glycan 828:835 arg1 structures 837:846 the N-glycan structures 824:846 the N-glycan structures using mass spectra (MS) analysis 824:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 3 42 theme sialic 680:685 arg1 content 692:698 the sialic acid content 676:698 the sialic acid content of total glycans on EPO 676:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 1 43 theme circulatory 250:260 arg1 lifetime 262:269 circulatory lifetime 250:269 circulatory lifetime 250:269 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 7 44 theme wild-type 1491:1499 arg1 CHO-K1 1510:1515 the wild-type parental CHO-K1 1487:1515 the wild-type parental CHO-K1 1487:1515 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 6 45 theme total 1239:1243 arg1 N-glycans 1245:1253 the total N-glycans 1235:1253 the total N-glycans on the resulting EPO 1235:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 8 46 theme tailored 1752:1759 arg1 structures 1778:1787 tailored complex N-glycan structures 1752:1787 tailored complex N-glycan structures on therapeutic glycoproteins in the future 1752:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 4 47 theme detailed 802:809 arg1 profiling 811:819 detailed profiling 802:819 detailed profiling of the N-glycan structures using mass spectra (MS) analysis 802:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 3 48 theme total 703:707 arg1 glycans 709:715 total glycans 703:715 total glycans on EPO 703:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 8 49 theme N-glycan 1769:1776 arg1 structures 1778:1787 tailored complex N-glycan structures 1752:1787 tailored complex N-glycan structures on therapeutic glycoproteins in the future 1752:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 7 50 theme chains 1460:1465 arg1 chains 1460:1465 total sugar chains 1448:1465 total sugar chains 1448:1465 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 7 50 theme chains 1460:1465 arg1 % 1443:1443 ∼10% 1440:1443 ∼10% of total sugar chains 1440:1465 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 3 51 from glycans 709:715 arg1 EPO 720:722 EPO 720:722 EPO 720:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 7 52 theme total 1448:1452 arg1 chains 1460:1465 total sugar chains 1448:1465 total sugar chains 1448:1465 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 3 53 theme cellular 589:596 arg1 proteins 598:605 CHO cellular proteins 585:605 CHO cellular proteins 585:605 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 1 54 theme key 204:206 arg1 roles 208:212 key roles 204:212 key roles 204:212 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 4 55 theme spectra 859:865 arg1 analysis 872:879 mass spectra (MS) analysis 854:879 mass spectra (MS) analysis 854:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 1 56 gly glycoproteins 286:298 arg1 glycoproteins 286:298 recombinant glycoproteins 274:298 recombinant glycoproteins of therapeutic interest 274:322 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 1 57 theme terminal 127:134 arg1 residue 136:142 a terminal residue 125:142 a terminal residue on complex N-glycans 125:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 4 58 theme MS 868:869 arg1 analysis 872:879 mass spectra (MS) analysis 854:879 mass spectra (MS) analysis 854:879 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 1 59 dep activity 237:244 arg1 the 222:224 the 222:224 the 222:224 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 6 60 theme resulting 1262:1270 arg1 EPO 1272:1274 the resulting EPO 1258:1274 the resulting EPO 1258:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 0 61 theme N-glycan 76:83 arg1 branching 85:93 enhanced erythropoietin N-glycan branching 52:93 enhanced erythropoietin N-glycan branching 52:93 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 62 from antennarity 183:193 arg1 N-glycans 155:163 complex N-glycans 147:163 complex N-glycans 147:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 8 63 theme branching 1684:1692 arg1 complexity 1694:1703 N-glycan branching complexity 1675:1703 N-glycan branching complexity 1675:1703 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 1 64 from branching 170:178 arg1 N-glycans 155:163 complex N-glycans 147:163 complex N-glycans 147:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 6 65 from EPO 1272:1274 arg1 % 1230:1230 approximately 92% 1214:1230 approximately 92% of the total N-glycans on the resulting EPO 1214:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 6 65 from EPO 1272:1274 arg1 N-glycans 1245:1253 the total N-glycans 1235:1253 the total N-glycans on the resulting EPO 1235:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 7 66 theme acid 1327:1330 arg1 content 1332:1338 sialic acid content 1320:1338 sialic acid content of rEPO from these engineered cells 1320:1374 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 5 67 theme α-1,3-D-mannoside 950:966 arg1 GnTIV/Mgat4 1006:1016 GnTIV/Mgat4 1006:1016 GnTIV/Mgat4 1006:1016 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 67 theme α-1,3-D-mannoside 950:966 arg1 β1,4-N-acetylglucosaminyltransferase 968:1003 α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase 950:1003 α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) 950:1017 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 7 68 theme increased 1380:1388 arg1 % 1393:1393 increased ∼45% 1380:1393 increased ∼45% 1380:1393 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 2 69 theme hamster 517:523 arg1 cells 540:544 Chinese hamster ovary (CHO-K1) cells 509:544 Chinese hamster ovary (CHO-K1) cells 509:544 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 8 70 theme glycosyltransferases 1573:1592 arg1 overexpression 1543:1556 coordinated overexpression 1531:1556 coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines 1531:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 6 71 from N-glycans 1245:1253 arg1 EPO 1272:1274 the resulting EPO 1258:1274 the resulting EPO 1258:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 1 72 from residue 136:142 arg1 N-glycans 155:163 complex N-glycans 147:163 complex N-glycans 147:163 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 3 73 from EPO 720:722 arg1 content 692:698 the sialic acid content 676:698 the sialic acid content of total glycans on EPO 676:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 0 74 theme hamster 28:34 arg1 cells 42:46 Chinese hamster ovary cells 20:46 Chinese hamster ovary cells 20:46 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 75 theme recombinant 274:284 arg1 glycoproteins 286:298 recombinant glycoproteins 274:298 recombinant glycoproteins of therapeutic interest 274:322 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 1 76 theme Sialic 112:117 arg1 acid 119:122 Sialic acid 112:122 Sialic acid 112:122 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 6 77 from % 1230:1230 arg1 EPO 1272:1274 the resulting EPO 1258:1274 the resulting EPO 1258:1274 Tri- and tetraantennary N-glycans represented approximately 92% of the total N-glycans on the resulting EPO as measured using MS analysis. 26154505 0 78 theme cells 42:46 arg1 Glycoengineering 0:15 Glycoengineering 0:15 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 0:110 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 7 79 theme rEPO 1343:1346 arg1 content 1332:1338 sialic acid content 1320:1338 sialic acid content of rEPO from these engineered cells 1320:1374 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 8 80 theme model 1616:1620 arg1 lines 1634:1638 model CHO-K1 cell lines 1616:1638 model CHO-K1 cell lines 1616:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 3 81 theme lectin 626:631 arg1 analysis 633:640 SNA lectin analysis 622:640 SNA lectin analysis 622:640 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 2 82 theme recombinant 416:426 arg1 erythropoietin 428:441 recombinant erythropoietin 416:441 recombinant erythropoietin (rEPO) 416:448 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 2 82 theme recombinant 416:426 arg1 rEPO 444:447 rEPO 444:447 rEPO 444:447 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 0 83 theme enhanced 52:59 arg1 branching 85:93 enhanced erythropoietin N-glycan branching 52:93 enhanced erythropoietin N-glycan branching 52:93 Glycoengineering of Chinese hamster ovary cells for enhanced erythropoietin N-glycan branching and sialylation. 26154505 1 84 theme interest 315:322 arg1 glycoproteins 286:298 recombinant glycoproteins 274:298 recombinant glycoproteins of therapeutic interest 274:322 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 26154505 5 85 theme α-1,6-D-mannoside 1047:1063 arg1 GnTV/Mgat5 1103:1112 GnTV/Mgat5 1103:1112 GnTV/Mgat5 1103:1112 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 5 85 theme α-1,6-D-mannoside 1047:1063 arg1 β1,6-N-acetylglucosaminyltransferase 1065:1100 α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase 1047:1100 α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5) 1047:1113 In order to enhance antennarity/branching, UDP-N-acetylglucosamine: α-1,3-D-mannoside β1,4-N-acetylglucosaminyltransferase (GnTIV/Mgat4) and UDP-N-acetylglucosamine:α-1,6-D-mannoside β1,6-N-acetylglucosaminyltransferase (GnTV/Mgat5), was incorporated into CHO-K1 together with ST6Gal1. 26154505 8 86 theme cell 1629:1632 arg1 lines 1634:1638 model CHO-K1 cell lines 1616:1638 model CHO-K1 cell lines 1616:1638 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 7 87 theme sialic 1320:1325 arg1 content 1332:1338 sialic acid content 1320:1338 sialic acid content of rEPO from these engineered cells 1320:1374 Furthermore, sialic acid content of rEPO from these engineered cells was increased ∼45% higher with tetra-sialylation accounting for ∼10% of total sugar chains compared to ∼3% for the wild-type parental CHO-K1. 26154505 3 88 theme HPLC 647:650 arg1 profiling 652:660 HPLC profiling 647:660 HPLC profiling 647:660 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 8 89 from glycoproteins 1804:1816 arg1 future 1825:1830 future 1825:1830 future 1825:1830 In this way, coordinated overexpression of these three glycosyltransferases for the first time in model CHO-K1 cell lines provides a mean for enhancing both N-glycan branching complexity and sialylation with opportunities to generate tailored complex N-glycan structures on therapeutic glycoproteins in the future. 26154505 3 90 theme acid 687:690 arg1 content 692:698 the sialic acid content 676:698 the sialic acid content of total glycans on EPO 676:722 Sialylation increased on both EPO and CHO cellular proteins as observed by SNA lectin analysis, and HPLC profiling revealed that the sialic acid content of total glycans on EPO increased by 26%. 26154505 4 91 theme acid 765:768 arg1 content 770:776 sialic acid content 758:776 sialic acid content 758:776 The increase in sialic acid content was further verified by detailed profiling of the N-glycan structures using mass spectra (MS) analysis. 26154505 2 92 theme glycosyltransferase 359:377 arg1 expression 379:388 glycosyltransferase expression 359:388 glycosyltransferase expression 359:388 In order to examine the impact of glycosyltransferase expression on the N-glycosylation of recombinant erythropoietin (rEPO), a human α2,6-sialyltransferase (ST6Gal1) was expressed in Chinese hamster ovary (CHO-K1) cells. 26154505 1 93 theme biological 226:235 arg1 activity 237:244 biological activity 226:244 biological activity 226:244 Sialic acid, a terminal residue on complex N-glycans, and branching or antennarity can play key roles in both the biological activity and circulatory lifetime of recombinant glycoproteins of therapeutic interest. 25598335 4 0 theme technology 507:516 arg1 matures 518:524 the technology matures 503:524 the technology matures 503:524 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 3 1 from glycans 458:464 arg1 proteins 481:488 recombinant proteins 469:488 recombinant proteins 469:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 4 2 dep come 549:552 arg1 up 554:555 up 554:555 up 554:555 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 5 3 from N- 794:795 arg1 yeasts 832:837 yeasts 832:837 yeasts of industrial importance 832:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 4 4 theme relevant 702:709 arg1 biopharmaceuticals 711:728 relevant biopharmaceuticals 702:728 relevant biopharmaceuticals 702:728 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 1 5 theme mammalian 112:120 arg1 culture 127:133 mammalian cell culture 112:133 mammalian cell culture 112:133 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 1 6 theme recombinant 157:167 arg1 glycoproteins 181:193 recombinant therapeutic glycoproteins 157:193 recombinant therapeutic glycoproteins 157:193 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 4 7 theme glycan 671:676 arg1 homogeneity 678:688 the unprecedented glycan homogeneity 653:688 the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals 653:728 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 3 8 theme great 340:344 arg1 value 346:350 great value 340:350 great value for therapeutic protein production and indispensable 340:403 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 1 9 gly glycoproteins 181:193 arg1 glycoproteins 181:193 recombinant therapeutic glycoproteins 157:193 recombinant therapeutic glycoproteins 157:193 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 2 10 theme N-glycosylation 255:269 arg1 pathway 271:277 the human N-glycosylation pathway 245:277 the human N-glycosylation pathway 245:277 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 3 11 theme glycans 458:464 arg1 relationships 441:453 the structure-function relationships 418:453 the structure-function relationships of glycans on recombinant proteins 418:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 5 12 dep N- 794:795 arg1 engineering 817:827 engineering 817:827 engineering 817:827 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 2 13 theme human 306:310 arg1 O-glycosylation 312:326 human O-glycosylation 306:326 human O-glycosylation 306:326 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 3 14 theme structure-function 422:439 arg1 relationships 441:453 the structure-function relationships 418:453 the structure-function relationships of glycans on recombinant proteins 418:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 5 15 theme recombinant 876:886 arg1 therapeutics 888:899 recombinant therapeutics 876:899 recombinant therapeutics with customized glycans 876:923 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 16 from developments 778:789 arg1 O-glycosylation 801:815 O-glycosylation 801:815 O-glycosylation 801:815 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 16 from developments 778:789 arg1 N- 794:795 N- 794:795 N- 794:795 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 1 17 theme therapeutic 169:179 arg1 glycoproteins 181:193 recombinant therapeutic glycoproteins 157:193 recombinant therapeutic glycoproteins 157:193 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 3 18 theme therapeutic 356:366 arg1 production 376:385 therapeutic protein production 356:385 therapeutic protein production 356:385 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 1 19 theme glycoproteins 181:193 arg1 production 143:152 the production 139:152 the production of recombinant therapeutic glycoproteins 139:193 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 3 20 theme protein 368:374 arg1 production 376:385 therapeutic protein production 356:385 therapeutic protein production 356:385 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 4 21 theme biomedical 573:582 arg1 applications 584:595 biomedical applications 573:595 biomedical applications 573:595 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 2 22 theme specific 288:295 arg1 types 297:301 specific types 288:301 specific types of human O-glycosylation 288:326 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 4 23 theme full 635:638 arg1 potential 640:648 the full potential 631:648 the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals 631:728 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 5 24 theme special 739:745 arg1 report 747:752 this special report 734:752 this special report 734:752 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 4 25 theme homogeneity 678:688 arg1 potential 640:648 the full potential 631:648 the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals 631:728 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 0 26 theme human 32:36 arg1 glycoproteins 38:50 human glycoproteins: 32:51 human glycoproteins: where are we now 32:68 Engineering yeast for producing human glycoproteins: where are we now? 25598335 3 27 theme recombinant 469:479 arg1 proteins 481:488 recombinant proteins 469:488 recombinant proteins 469:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 5 28 theme industrial 842:851 arg1 importance 853:862 industrial importance 842:862 industrial importance 842:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 2 29 theme yeast 207:211 arg1 strains 213:219 Engineered yeast strains 196:219 Engineered yeast strains 196:219 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 3 30 from proteins 481:488 arg1 relationships 441:453 the structure-function relationships 418:453 the structure-function relationships of glycans on recombinant proteins 418:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 2 31 theme Engineered 196:205 arg1 strains 213:219 Engineered yeast strains 196:219 Engineered yeast strains 196:219 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 2 32 theme human 249:253 arg1 pathway 271:277 the human N-glycosylation pathway 245:277 the human N-glycosylation pathway 245:277 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 5 33 theme importance 853:862 arg1 yeasts 832:837 yeasts 832:837 yeasts of industrial importance 832:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 34 theme customized 906:915 arg1 glycans 917:923 customized glycans 906:923 customized glycans 906:923 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 35 with therapeutics 888:899 arg1 glycans 917:923 customized glycans 906:923 customized glycans 906:923 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 0 36 gly glycoproteins 38:50 arg1 glycoproteins 38:50 human glycoproteins: 32:51 human glycoproteins: where are we now 32:68 Engineering yeast for producing human glycoproteins: where are we now? 25598335 2 37 theme O-glycosylation 312:326 arg1 types 297:301 specific types 288:301 specific types of human O-glycosylation 288:326 Engineered yeast strains not only allow to mimic the human N-glycosylation pathway but also specific types of human O-glycosylation. 25598335 3 38 from relationships 441:453 arg1 proteins 481:488 recombinant proteins 469:488 recombinant proteins 469:488 This is of great value for therapeutic protein production and indispensable to determine the structure-function relationships of glycans on recombinant proteins. 25598335 1 39 theme cell 122:125 arg1 culture 127:133 mammalian cell culture 112:133 mammalian cell culture 112:133 Yeast has advanced as an alternative for mammalian cell culture for the production of recombinant therapeutic glycoproteins. 25598335 4 40 theme unprecedented 657:669 arg1 homogeneity 678:688 the unprecedented glycan homogeneity 653:688 the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals 653:728 However, as the technology matures, some limitations have come up that may hamper biomedical applications and must be considered to exploit the full potential of the unprecedented glycan homogeneity obtained on relevant biopharmaceuticals. 25598335 5 41 theme recent 771:776 arg1 developments 778:789 the recent developments 767:789 the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance 767:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 42 from O-glycosylation 801:815 arg1 yeasts 832:837 yeasts 832:837 yeasts of industrial importance 832:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 25598335 5 43 gly O-glycosylation 801:815 arg1 yeasts 832:837 yeasts 832:837 yeasts of industrial importance 832:862 In this special report, we focus on the recent developments in N- and O-glycosylation engineering in yeasts of industrial importance, to produce recombinant therapeutics with customized glycans. 26101185 5 0 theme EndoBI-1 821:828 arg1 activity 830:837 EndoBI-1 activity 821:837 EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry 821:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 7 1 theme reaction 1058:1065 arg1 conditions 1067:1076 different reaction conditions 1048:1076 different reaction conditions 1048:1076 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 1 2 theme different 184:192 arg1 signaling 237:245 signaling 237:245 signaling 237:245 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 2 theme different 184:192 arg1 processes 203:211 different cellular processes 184:211 different cellular processes 184:211 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 2 theme different 184:192 arg1 adhesion 224:231 adhesion 224:231 adhesion 224:231 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 4 3 theme heat-stable 576:586 arg1 15697 547:551 infantis ATCC 15697 533:551 infantis ATCC 15697 533:551 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 4 3 theme heat-stable 576:586 arg1 enzyme 588:593 a recently isolated heat-stable enzyme 556:593 a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core 556:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 7 4 theme N-glycan 1173:1180 arg1 types 1182:1186 N-glycan types 1173:1186 N-glycan types 1173:1186 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 1 5 theme cellular 194:201 arg1 signaling 237:245 signaling 237:245 signaling 237:245 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 5 theme cellular 194:201 arg1 processes 203:211 different cellular processes 184:211 different cellular processes 184:211 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 5 theme cellular 194:201 arg1 adhesion 224:231 adhesion 224:231 adhesion 224:231 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 6 theme infant 281:286 arg1 microbiome 288:297 the infant microbiome 277:297 the infant microbiome 277:297 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 1 7 gly glycoproteins 116:128 arg1 glycoproteins 116:128 Milk glycoproteins 111:128 Milk glycoproteins 111:128 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 4 8 theme isolated 567:574 arg1 15697 547:551 infantis ATCC 15697 533:551 infantis ATCC 15697 533:551 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 4 8 theme isolated 567:574 arg1 enzyme 588:593 a recently isolated heat-stable enzyme 556:593 a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core 556:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 1 9 theme microbiome 288:297 arg1 development 262:272 the development 258:272 the development of the infant microbiome 258:297 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 8 10 theme lower 1244:1248 arg1 temperatures 1250:1261 lower temperatures 1244:1261 lower temperatures 1244:1261 In general, more sialylated N-glycans were released at lower temperatures and pH values. 26101185 4 11 theme ATCC 542:545 arg1 15697 547:551 infantis ATCC 15697 533:551 infantis ATCC 15697 533:551 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 4 11 theme ATCC 542:545 arg1 enzyme 588:593 a recently isolated heat-stable enzyme 556:593 a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core 556:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 9 12 theme different 1423:1431 arg1 conditions 1444:1453 different processing conditions 1423:1453 different processing conditions 1423:1453 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 4 13 theme N-glycan 657:664 arg1 core 666:669 the N-glycan core 653:669 the N-glycan core 653:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 8 14 theme pH 1267:1268 arg1 values 1270:1275 pH values 1267:1275 pH values 1267:1275 In general, more sialylated N-glycans were released at lower temperatures and pH values. 26101185 4 15 theme infantis 533:540 arg1 15697 547:551 infantis ATCC 15697 533:551 infantis ATCC 15697 533:551 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 4 15 theme infantis 533:540 arg1 enzyme 588:593 a recently isolated heat-stable enzyme 556:593 a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core 556:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 5 16 theme different 687:695 arg1 conditions 708:717 different processing conditions 687:717 different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 687:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 16 theme different 687:695 arg1 ratio 771:775 enzyme/protein ratio 756:775 enzyme/protein ratio 756:775 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 16 theme different 687:695 arg1 temperature 724:734 temperature 724:734 temperature 724:734 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 16 theme different 687:695 arg1 time 746:749 reaction time 737:749 reaction time 737:749 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 16 theme different 687:695 arg1 pH 720:721 pH 720:721 pH 720:721 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 2 17 theme protein 424:430 arg1 functionality 432:444 protein functionality 424:444 protein functionality 424:444 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 5 18 theme processing 697:706 arg1 conditions 708:717 different processing conditions 687:717 different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 687:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 18 theme processing 697:706 arg1 ratio 771:775 enzyme/protein ratio 756:775 enzyme/protein ratio 756:775 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 18 theme processing 697:706 arg1 temperature 724:734 temperature 724:734 temperature 724:734 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 18 theme processing 697:706 arg1 time 746:749 reaction time 737:749 reaction time 737:749 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 18 theme processing 697:706 arg1 pH 720:721 pH 720:721 pH 720:721 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 7 19 theme Nano-LC-Chip-Q-TOF 1002:1019 arg1 data 1024:1027 Nano-LC-Chip-Q-TOF MS data 1002:1027 Nano-LC-Chip-Q-TOF MS data 1002:1027 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 4 20 located found 644:648 arg1 core 666:669 the N-glycan core 653:669 the N-glycan core 653:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 4 20 located found 644:648 arg2 moiety 637:642 the N-N'-diacetyl chitobiose moiety 608:642 the N-N'-diacetyl chitobiose moiety found in the N-glycan core 608:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 0 21 theme novel 2:6 arg1 endo-β-N-acetylglucosaminidase 8:37 A novel endo-β-N-acetylglucosaminidase 0:37 A novel endo-β-N-acetylglucosaminidase 0:37 A novel endo-β-N-acetylglucosaminidase releases specific N-glycans depending on different reaction conditions. 26101185 5 22 theme conditions 708:717 arg1 effects 676:682 The effects 672:682 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 672:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 7 23 theme MS 1021:1022 arg1 data 1024:1027 Nano-LC-Chip-Q-TOF MS data 1002:1027 Nano-LC-Chip-Q-TOF MS data 1002:1027 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 1 24 theme Milk 111:114 arg1 glycoproteins 116:128 Milk glycoproteins 111:128 Milk glycoproteins 111:128 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 2 25 theme N-glycans 411:419 arg1 role 403:406 the role 399:406 the role of N-glycans in protein functionality 399:444 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 4 26 theme chitobiose 626:635 arg1 moiety 637:642 the N-N'-diacetyl chitobiose moiety 608:642 the N-N'-diacetyl chitobiose moiety found in the N-glycan core 608:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 7 27 theme types 1182:1186 arg1 abundance 1160:1168 the relative abundance 1147:1168 the relative abundance of N-glycan types 1147:1186 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 2 28 from role 403:406 arg1 functionality 432:444 protein functionality 424:444 protein functionality 424:444 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 5 29 theme advanced 884:891 arg1 spectrometry 898:909 advanced mass spectrometry 884:909 advanced mass spectrometry 884:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 4 30 theme N-N'-diacetyl 612:624 arg1 moiety 637:642 the N-N'-diacetyl chitobiose moiety 608:642 the N-N'-diacetyl chitobiose moiety found in the N-glycan core 608:669 infantis ATCC 15697 is a recently isolated heat-stable enzyme that cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core. 26101185 5 31 theme enzyme/protein 756:769 arg1 conditions 708:717 different processing conditions 687:717 different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 687:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 31 theme enzyme/protein 756:769 arg1 ratio 771:775 enzyme/protein ratio 756:775 enzyme/protein ratio 756:775 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 32 theme mass 893:896 arg1 spectrometry 898:909 advanced mass spectrometry 884:909 advanced mass spectrometry 884:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 33 dep conditions 708:717 arg1 conditions 708:717 different processing conditions 687:717 different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 687:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 33 dep conditions 708:717 arg1 ratio 771:775 enzyme/protein ratio 756:775 enzyme/protein ratio 756:775 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 33 dep conditions 708:717 arg1 temperature 724:734 temperature 724:734 temperature 724:734 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 33 dep conditions 708:717 arg1 time 746:749 reaction time 737:749 reaction time 737:749 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 33 dep conditions 708:717 arg1 pH 720:721 pH 720:721 pH 720:721 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 2 34 theme protein 362:368 arg1 glycosylation 370:382 milk protein glycosylation 357:382 milk protein glycosylation 357:382 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 8 35 gly sialylated 1206:1215 arg1 N-glycans 1217:1225 more sialylated N-glycans 1201:1225 more sialylated N-glycans 1201:1225 In general, more sialylated N-glycans were released at lower temperatures and pH values. 26101185 0 36 theme specific 48:55 arg1 N-glycans 57:65 specific N-glycans 48:65 specific N-glycans 48:65 A novel endo-β-N-acetylglucosaminidase releases specific N-glycans depending on different reaction conditions. 26101185 5 37 theme bovine 842:847 arg1 glycoproteins 864:876 bovine colostrum whey glycoproteins 842:876 bovine colostrum whey glycoproteins using advanced mass spectrometry 842:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 38 gly glycoproteins 864:876 arg1 glycoproteins 864:876 bovine colostrum whey glycoproteins 842:876 bovine colostrum whey glycoproteins using advanced mass spectrometry 842:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 7 39 theme N-glycan 1100:1107 arg1 compositions 1109:1120 different N-glycan compositions 1090:1120 different N-glycan compositions released 1090:1129 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 3 40 theme longum 519:524 arg1 subsp 526:530 Bifidobacterium longum subsp 503:530 Bifidobacterium longum subsp 503:530 Endo-β-N-acetylglucosaminidase (EndoBI-1) isolated from Bifidobacterium longum subsp. 26101185 5 41 theme colostrum 849:857 arg1 glycoproteins 864:876 bovine colostrum whey glycoproteins 842:876 bovine colostrum whey glycoproteins using advanced mass spectrometry 842:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 42 theme reaction 737:744 arg1 conditions 708:717 different processing conditions 687:717 different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) 687:776 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 5 42 theme reaction 737:744 arg1 time 746:749 reaction time 737:749 reaction time 737:749 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 9 43 theme processing 1433:1442 arg1 conditions 1444:1453 different processing conditions 1423:1453 different processing conditions 1423:1453 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 2 44 theme glycosylation 370:382 arg1 complexities 341:352 the complexities 337:352 the complexities of milk protein glycosylation 337:382 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 5 45 theme whey 859:862 arg1 glycoproteins 864:876 bovine colostrum whey glycoproteins 842:876 bovine colostrum whey glycoproteins using advanced mass spectrometry 842:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 9 46 theme wide 1340:1343 arg1 variety 1345:1351 a wide variety 1338:1351 a wide variety 1338:1351 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 9 46 theme wide 1340:1343 arg1 N-glycans 1356:1364 N-glycans 1356:1364 N-glycans 1356:1364 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 1 47 theme different 146:154 arg1 functions 156:164 different functions 146:164 different functions 146:164 Milk glycoproteins are involved in different functions and contribute to different cellular processes, including adhesion and signaling, and shape the development of the infant microbiome. 26101185 6 48 theme structures 990:999 arg1 diversity 968:976 a high diversity 961:976 a high diversity of N-glycan structures 961:999 This study shows that EndoBI-1 is able to cleave a high diversity of N-glycan structures. 26101185 8 49 theme sialylated 1206:1215 arg1 N-glycans 1217:1225 more sialylated N-glycans 1201:1225 more sialylated N-glycans 1201:1225 In general, more sialylated N-glycans were released at lower temperatures and pH values. 26101185 5 50 from activity 830:837 arg1 glycoproteins 864:876 bovine colostrum whey glycoproteins 842:876 bovine colostrum whey glycoproteins using advanced mass spectrometry 842:909 The effects of different processing conditions (pH, temperature, reaction time, and enzyme/protein ratio) were evaluated for their ability to change EndoBI-1 activity on bovine colostrum whey glycoproteins using advanced mass spectrometry. 26101185 7 51 theme different 1090:1098 arg1 compositions 1109:1120 different N-glycan compositions 1090:1120 different N-glycan compositions released 1090:1129 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 2 52 theme milk 357:360 arg1 glycosylation 370:382 milk protein glycosylation 357:382 milk protein glycosylation 357:382 Methods have been developed to study the complexities of milk protein glycosylation and understand the role of N-glycans in protein functionality. 26101185 0 53 theme reaction 90:97 arg1 conditions 99:108 different reaction conditions 80:108 different reaction conditions 80:108 A novel endo-β-N-acetylglucosaminidase releases specific N-glycans depending on different reaction conditions. 26101185 9 54 theme N-glycans 1356:1364 arg1 variety 1345:1351 a wide variety 1338:1351 a wide variety 1338:1351 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 9 54 theme N-glycans 1356:1364 arg1 N-glycans 1356:1364 N-glycans 1356:1364 N-glycans 1356:1364 These results demonstrated that EndoBI-1 is able to release a wide variety of N-glycans, whose compositions can be selectively manipulated using different processing conditions. 26101185 7 55 theme relative 1151:1158 arg1 abundance 1160:1168 the relative abundance 1147:1168 the relative abundance of N-glycan types 1147:1186 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 26101185 6 56 theme high 963:966 arg1 diversity 968:976 a high diversity 961:976 a high diversity of N-glycan structures 961:999 This study shows that EndoBI-1 is able to cleave a high diversity of N-glycan structures. 26101185 0 57 theme different 80:88 arg1 conditions 99:108 different reaction conditions 80:108 different reaction conditions 80:108 A novel endo-β-N-acetylglucosaminidase releases specific N-glycans depending on different reaction conditions. 26101185 6 58 theme N-glycan 981:988 arg1 structures 990:999 N-glycan structures 981:999 N-glycan structures 981:999 This study shows that EndoBI-1 is able to cleave a high diversity of N-glycan structures. 26101185 7 59 theme different 1048:1056 arg1 conditions 1067:1076 different reaction conditions 1048:1076 different reaction conditions 1048:1076 Nano-LC-Chip-Q-TOF MS data also revealed that different reaction conditions resulted in different N-glycan compositions released, thus modifying the relative abundance of N-glycan types. 24519758 8 0 theme goat 1123:1126 arg1 milk 1128:1131 goat milk 1123:1131 goat milk that were not present in human milk 1123:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 5 1 theme advanced 699:706 arg1 MS. 708:710 advanced MS. 699:710 advanced MS. 699:710 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 7 2 gly sialylated 968:977 arg1 % 961:961 37% 959:961 37% 959:961 Among the N-glycan compositions, 37% were sialylated and 34% were fucosylated. 24519758 4 3 theme lactoferrin 506:516 arg1 pattern 485:491 the glycosylation pattern 467:491 the glycosylation pattern of goat milk lactoferrin 467:516 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 5 4 theme glycan 661:666 arg1 composition 668:678 glycan composition 661:678 glycan composition 661:678 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 6 5 theme MS 837:838 arg1 data 840:843 Nano-LC-Chip-Q-TOF MS data 818:843 Nano-LC-Chip-Q-TOF MS data 818:843 Nano-LC-Chip-Q-TOF MS data identified 65 structures, including high mannose, hybrid, and complex N-glycans. 24519758 5 6 theme goat 733:736 arg1 lactoferrin 743:753 goat milk lactoferrin 733:753 goat milk lactoferrin 733:753 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 4 7 theme milk 501:504 arg1 lactoferrin 506:516 goat milk lactoferrin 496:516 goat milk lactoferrin 496:516 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 5 8 theme composition 668:678 arg1 characterization 641:656 a complete characterization 630:656 a complete characterization of glycan composition 630:678 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 6 9 theme Nano-LC-Chip-Q-TOF 818:835 arg1 data 840:843 Nano-LC-Chip-Q-TOF MS data 818:843 Nano-LC-Chip-Q-TOF MS data 818:843 Nano-LC-Chip-Q-TOF MS data identified 65 structures, including high mannose, hybrid, and complex N-glycans. 24519758 0 10 theme human 90:94 arg1 N-glycomes 76:85 the N-glycomes 72:85 the N-glycomes of human and bovine milk 72:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 3 11 theme infant 337:342 arg1 supplementation 352:366 infant formula supplementation 337:366 infant formula supplementation 337:366 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 8 12 theme glycans 1055:1061 arg1 existence 1034:1042 the existence 1030:1042 the existence of similar glycans in human and goat milk 1030:1084 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 0 13 theme bovine 100:105 arg1 N-glycomes 76:85 the N-glycomes 72:85 the N-glycomes of human and bovine milk 72:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 0 14 theme N-glycans 42:50 arg1 Characterization 0:15 Characterization 0:15 Characterization of goat milk lactoferrin N-glycans 0:50 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 0 14 theme N-glycans 42:50 arg1 comparison 56:65 comparison 56:65 comparison with the N-glycomes of human and bovine milk 56:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 0 15 dep human 90:94 arg1 milk 107:110 milk 107:110 milk 107:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 5 16 theme lactoferrin 743:753 arg1 glycosylation 716:728 The glycosylation 712:728 The glycosylation of goat milk lactoferrin 712:753 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 3 17 theme high 383:386 arg1 homology 388:395 its high homology 379:395 its high homology with its human counterpart 379:422 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 4 18 theme glycosylation 471:483 arg1 pattern 485:491 the glycosylation pattern 467:491 the glycosylation pattern of goat milk lactoferrin 467:516 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 5 19 theme complete 632:639 arg1 characterization 641:656 a complete characterization 630:656 a complete characterization of glycan composition 630:678 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 5 20 theme affinity 563:570 arg1 chromatography 572:585 affinity chromatography 563:585 affinity chromatography 563:585 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 8 21 theme human 1158:1162 arg1 milk 1164:1167 human milk 1158:1167 human milk 1158:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 9 22 theme functional 1290:1299 arg1 ingredients 1301:1311 functional ingredients 1290:1311 functional ingredients for food products beneficial to human nutrition 1290:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 22 theme functional 1290:1299 arg1 source 1215:1220 a source 1213:1220 a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition 1213:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 23 used used 1282:1285 arg2 milk 1199:1202 goat milk 1194:1202 goat milk 1194:1202 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 23 used used 1282:1285 arg2 source 1215:1220 a source 1213:1220 a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition 1213:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 23 used used 1282:1285 arg2 ingredients 1301:1311 functional ingredients 1290:1311 functional ingredients for food products beneficial to human nutrition 1290:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 0 24 theme milk 25:28 arg1 N-glycans 42:50 goat milk lactoferrin N-glycans 20:50 goat milk lactoferrin N-glycans 20:50 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 8 25 from existence 1034:1042 arg1 milk 1081:1084 goat milk 1076:1084 goat milk 1076:1084 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 8 25 from existence 1034:1042 arg1 human 1066:1070 human 1066:1070 human 1066:1070 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 5 26 theme human 781:785 arg1 glycoproteins 803:815 human and bovine milk glycoproteins 781:815 human and bovine milk glycoproteins 781:815 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 7 27 theme N-glycan 936:943 arg1 compositions 945:956 the N-glycan compositions 932:956 the N-glycan compositions 932:956 Among the N-glycan compositions, 37% were sialylated and 34% were fucosylated. 24519758 8 28 attach present 1147:1153 arg1 milk 1164:1167 human milk 1158:1167 human milk 1158:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 8 28 attach present 1147:1153 arg2 milk 1128:1131 goat milk 1123:1131 goat milk that were not present in human milk 1123:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 1 29 theme health-promoting 178:193 arg1 compounds 195:203 health-promoting compounds 178:203 health-promoting compounds 178:203 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 29 theme health-promoting 178:193 arg1 components 127:136 Numerous milk components 113:136 Numerous milk components 113:136 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 29 theme health-promoting 178:193 arg1 lactoferrin 147:157 lactoferrin 147:157 lactoferrin 147:157 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 0 30 theme goat 20:23 arg1 N-glycans 42:50 goat milk lactoferrin N-glycans 20:50 goat milk lactoferrin N-glycans 20:50 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 4 31 theme study 441:445 arg1 aim 429:431 The aim 425:431 The aim of this study 425:445 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 3 32 with homology 388:395 arg1 counterpart 412:422 its human counterpart 402:422 its human counterpart 402:422 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 4 33 gly glycosylation 471:483 arg1 lactoferrin 506:516 goat milk lactoferrin 496:516 goat milk lactoferrin 496:516 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 3 34 theme human 406:410 arg1 counterpart 412:422 its human counterpart 402:422 its human counterpart 402:422 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 9 35 theme food 1317:1320 arg1 products 1322:1329 food products 1317:1329 food products beneficial to human nutrition 1317:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 6 36 theme high 881:884 arg1 mannose 886:892 high mannose 881:892 high mannose 881:892 Nano-LC-Chip-Q-TOF MS data identified 65 structures, including high mannose, hybrid, and complex N-glycans. 24519758 2 37 theme evidence 224:231 arg1 body 216:219 A growing body 206:219 A growing body of evidence 206:231 A growing body of evidence suggests that glycans could mediate lactoferrin's bioactivity. 24519758 0 38 theme lactoferrin 30:40 arg1 N-glycans 42:50 goat milk lactoferrin N-glycans 20:50 goat milk lactoferrin N-glycans 20:50 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 9 39 theme human 1345:1349 arg1 nutrition 1351:1359 human nutrition 1345:1359 human nutrition 1345:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 5 40 gly glycoproteins 803:815 arg1 glycoproteins 803:815 human and bovine milk glycoproteins 781:815 human and bovine milk glycoproteins 781:815 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 8 41 theme similar 1047:1053 arg1 glycans 1055:1061 similar glycans 1047:1061 similar glycans 1047:1061 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 3 42 theme formula 344:350 arg1 supplementation 352:366 infant formula supplementation 337:366 infant formula supplementation 337:366 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 5 43 theme bovine 791:796 arg1 glycoproteins 803:815 human and bovine milk glycoproteins 781:815 human and bovine milk glycoproteins 781:815 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 9 44 theme beneficial 1331:1340 arg1 products 1322:1329 food products 1317:1329 food products beneficial to human nutrition 1317:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 3 45 theme milk 301:304 arg1 candidate 323:331 a candidate 321:331 a candidate for infant formula supplementation 321:366 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 3 45 theme milk 301:304 arg1 lactoferrin 306:316 Goat milk lactoferrin 296:316 Goat milk lactoferrin 296:316 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 9 46 theme goat 1194:1197 arg1 milk 1199:1202 goat milk 1194:1202 goat milk 1194:1202 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 46 theme goat 1194:1197 arg1 source 1215:1220 a source 1213:1220 a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition 1213:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 5 47 theme milk 798:801 arg1 glycoproteins 803:815 human and bovine milk glycoproteins 781:815 human and bovine milk glycoproteins 781:815 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 8 48 theme goat 1076:1079 arg1 milk 1081:1084 goat milk 1076:1084 goat milk 1076:1084 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 8 49 from milk 1164:1167 arg1 present 1147:1153 present 1147:1153 present 1147:1153 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 2 50 theme growing 208:214 arg1 body 216:219 A growing body 206:219 A growing body of evidence 206:231 A growing body of evidence suggests that glycans could mediate lactoferrin's bioactivity. 24519758 1 51 theme Numerous 113:120 arg1 compounds 195:203 health-promoting compounds 178:203 health-promoting compounds 178:203 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 51 theme Numerous 113:120 arg1 components 127:136 Numerous milk components 113:136 Numerous milk components 113:136 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 51 theme Numerous 113:120 arg1 lactoferrin 147:157 lactoferrin 147:157 lactoferrin 147:157 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 7 52 gly fucosylated 992:1002 arg1 % 985:985 34% 983:985 34% 983:985 Among the N-glycan compositions, 37% were sialylated and 34% were fucosylated. 24519758 8 53 from present 1147:1153 arg1 milk 1164:1167 human milk 1158:1167 human milk 1158:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 0 54 with Characterization 0:15 arg1 N-glycomes 76:85 the N-glycomes 72:85 the N-glycomes of human and bovine milk 72:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 1 55 theme milk 122:125 arg1 compounds 195:203 health-promoting compounds 178:203 health-promoting compounds 178:203 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 55 theme milk 122:125 arg1 components 127:136 Numerous milk components 113:136 Numerous milk components 113:136 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 1 55 theme milk 122:125 arg1 lactoferrin 147:157 lactoferrin 147:157 lactoferrin 147:157 Numerous milk components, such as lactoferrin, are recognized as health-promoting compounds. 24519758 8 56 located present 1147:1153 arg1 milk 1164:1167 human milk 1158:1167 human milk 1158:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 8 56 located present 1147:1153 arg2 milk 1128:1131 goat milk 1123:1131 goat milk that were not present in human milk 1123:1167 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 3 57 theme Goat 296:299 arg1 candidate 323:331 a candidate 321:331 a candidate for infant formula supplementation 321:366 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 3 57 theme Goat 296:299 arg1 lactoferrin 306:316 Goat milk lactoferrin 296:316 Goat milk lactoferrin 296:316 Goat milk lactoferrin is a candidate for infant formula supplementation because of its high homology with its human counterpart. 24519758 9 58 theme bioactive 1225:1233 arg1 lactoferrin 1256:1266 lactoferrin 1256:1266 lactoferrin 1256:1266 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 58 theme bioactive 1225:1233 arg1 compounds 1235:1243 bioactive compounds 1225:1243 bioactive compounds 1225:1243 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 0 59 with comparison 56:65 arg1 N-glycomes 76:85 the N-glycomes 72:85 the N-glycomes of human and bovine milk 72:110 Characterization of goat milk lactoferrin N-glycans and comparison with the N-glycomes of human and bovine milk. 24519758 5 60 gly glycosylation 716:728 arg1 lactoferrin 743:753 goat milk lactoferrin 733:753 goat milk lactoferrin 733:753 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 5 61 attach isolated 541:548 arg2 protein 529:535 the protein 525:535 the protein 525:535 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 5 61 attach isolated 541:548 arg1 milk 555:558 milk 555:558 milk 555:558 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 24519758 9 62 theme compounds 1235:1243 arg1 milk 1199:1202 goat milk 1194:1202 goat milk 1194:1202 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 62 theme compounds 1235:1243 arg1 ingredients 1301:1311 functional ingredients 1290:1311 functional ingredients for food products beneficial to human nutrition 1290:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 9 62 theme compounds 1235:1243 arg1 source 1215:1220 a source 1213:1220 a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition 1213:1359 These data suggest that goat milk could be a source of bioactive compounds, including lactoferrin that could be used as functional ingredients for food products beneficial to human nutrition. 24519758 6 63 theme complex 907:913 arg1 N-glycans 915:923 complex N-glycans 907:923 complex N-glycans 907:923 Nano-LC-Chip-Q-TOF MS data identified 65 structures, including high mannose, hybrid, and complex N-glycans. 24519758 4 64 theme goat 496:499 arg1 lactoferrin 506:516 goat milk lactoferrin 496:516 goat milk lactoferrin 496:516 The aim of this study was to characterize the glycosylation pattern of goat milk lactoferrin. 24519758 8 65 theme novel 1106:1110 arg1 glycans 1112:1118 novel glycans 1106:1118 novel glycans 1106:1118 The results demonstrated the existence of similar glycans in human and goat milk but also identified novel glycans in goat milk that were not present in human milk. 24519758 5 66 theme milk 738:741 arg1 lactoferrin 743:753 goat milk lactoferrin 733:753 goat milk lactoferrin 733:753 After the protein was isolated from milk by affinity chromatography, N-glycans were enzymatically released and a complete characterization of glycan composition was carried out by advanced MS. The glycosylation of goat milk lactoferrin was compared with that of human and bovine milk glycoproteins. 28855248 4 0 theme immature 799:806 arg1 SCs 808:810 immature SCs 799:810 immature SCs isolated from 8-day-old rats 799:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 3 1 theme oligosaccharides 656:671 arg1 complexity 642:651 complexity 642:651 complexity 642:651 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 1 theme oligosaccharides 656:671 arg1 degree 619:624 their sialylation degree 601:624 their sialylation degree (AC and BA) 601:636 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 5 2 from effect 890:895 arg1 FSH 900:902 FSH 900:902 FSH 900:902 Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. 28855248 10 3 theme high 1452:1455 arg1 mannose 1457:1463 high mannose and hybrid-type oligosaccharides 1452:1496 mannose 1457:1463 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 3 4 theme FSH 539:541 arg1 variants 565:572 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 2 5 with interaction 384:394 arg1 factors 409:415 gonadal factors 401:415 gonadal factors 401:415 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 6 6 theme Pro-αC 1075:1080 arg1 production 1082:1091 Pro-αC production 1075:1091 Pro-αC production 1075:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 3 7 theme Recombinant 521:531 arg1 rhFSH 544:548 rhFSH 544:548 rhFSH 544:548 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 7 theme Recombinant 521:531 arg1 FSH 539:541 Recombinant human FSH 521:541 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 11 8 from effect 1686:1691 arg1 regulation 1733:1742 the regulation 1729:1742 the regulation of inhibin production in the male gonad 1729:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 1 9 theme sexual 265:270 arg1 maturation 272:281 sexual maturation 265:281 sexual maturation 265:281 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 3 10 theme glycosylation 551:563 arg1 variants 565:572 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 1 11 theme structural 201:210 arg1 changes 227:233 important structural and functional changes 191:233 important structural and functional changes in Sertoli cells (SCs) during sexual maturation 191:281 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 1 12 theme hormone 118:124 arg1 composition 145:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 8 13 theme growth 1325:1330 arg1 factors 1332:1338 growth factors 1325:1338 growth factors 1325:1338 In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. 28855248 11 14 theme structure 1716:1724 arg1 effect 1686:1691 the modulatory effect 1671:1691 the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad 1671:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 1 15 theme functional 216:225 arg1 changes 227:233 important structural and functional changes 191:233 important structural and functional changes in Sertoli cells (SCs) during sexual maturation 191:281 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 4 16 theme 8-day-old 826:834 arg1 rats 836:839 8-day-old rats 826:839 8-day-old rats 826:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 10 17 theme HY 1499:1500 arg1 Glycoforms 1433:1442 Glycoforms 1433:1442 Glycoforms bearing high mannose and hybrid-type oligosaccharides 1433:1496 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 11 18 theme FSH 1696:1698 arg1 structure 1716:1724 FSH oligosaccharide structure 1696:1724 FSH oligosaccharide structure 1696:1724 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 9 19 theme marked 1375:1380 arg1 effect 1394:1399 a more marked stimulatory effect 1368:1399 a more marked stimulatory effect on INHB and Pro-αC than the AC 1368:1430 The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. 28855248 2 20 from stages 503:508 arg1 regulation 424:433 the regulation 420:433 the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC 420:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 4 21 theme Native 686:691 arg1 rhFSH 693:697 Native rhFSH 686:697 Native rhFSH 686:697 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 10 22 theme factors 1635:1641 arg1 presence 1608:1615 the presence 1604:1615 the presence of gonadal growth factors 1604:1641 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 6 23 theme rhFSH 952:956 arg1 BA 976:977 BA 976:977 BA 976:977 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 23 theme rhFSH 952:956 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 23 theme rhFSH 952:956 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 2 24 theme maturation 492:501 arg1 stages 503:508 different maturation stages 482:508 different maturation stages of the SC 482:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 1 25 theme carbohydrate 132:143 arg1 composition 145:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 0 26 theme cell 58:61 arg1 production 71:80 Sertoli cell inhibin production 50:80 Sertoli cell inhibin production 50:80 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 2 27 theme SC 517:518 arg1 stages 503:508 different maturation stages 482:508 different maturation stages of the SC 482:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 0 28 from Role 0:3 arg1 regulation 36:45 the regulation 32:45 the regulation of Sertoli cell inhibin production 32:80 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 0 29 theme production 71:80 arg1 regulation 36:45 the regulation 32:45 the regulation of Sertoli cell inhibin production 32:80 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 4 30 from production 785:794 arg1 SCs 808:810 immature SCs 799:810 immature SCs isolated from 8-day-old rats 799:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 10 31 theme growth 1628:1633 arg1 factors 1635:1641 gonadal growth factors 1620:1641 gonadal growth factors 1620:1641 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 8 32 theme glycosylation 1240:1252 arg1 variants 1254:1261 all rhFSH glycosylation variants 1230:1261 all rhFSH glycosylation variants 1230:1261 In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. 28855248 6 33 theme only 989:992 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 33 theme only 989:992 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 11 34 from regulation 1733:1742 arg1 gonad 1778:1782 the male gonad 1769:1782 the male gonad 1769:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 2 35 theme structure 366:374 arg1 impact 336:341 the impact 332:341 the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC 332:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 3 36 dep oligosaccharides 656:671 arg1 CO 674:675 CO 674:675 CO 674:675 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 36 dep oligosaccharides 656:671 arg1 oligosaccharides 656:671 oligosaccharides 656:671 oligosaccharides (CO and HY) 656:683 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 36 dep oligosaccharides 656:671 arg1 HY 681:682 HY 681:682 HY 681:682 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 10 37 theme complex 1566:1572 arg1 oligosaccharides 1574:1589 complex oligosaccharides 1566:1589 complex oligosaccharides 1566:1589 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 0 38 theme FSH 8:10 arg1 structure 19:27 FSH glycan structure 8:27 FSH glycan structure 8:27 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 5 39 theme synergistic 878:888 arg1 effect 890:895 a synergistic effect 876:895 a synergistic effect on FSH to stimulate Pro-αC 876:922 Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. 28855248 2 40 theme inhibin 460:466 arg1 production 468:477 monomeric and dimeric inhibin production 438:477 monomeric and dimeric inhibin production 438:477 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 0 41 theme structure 19:27 arg1 Role 0:3 Role 0:3 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 0:81 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 7 42 attach isolated 1183:1190 arg2 SCs 1142:1144 SCs 1142:1144 SCs 1142:1144 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 7 42 attach isolated 1183:1190 arg1 rats 1208:1211 20-day-old rats 1197:1211 20-day-old rats 1197:1211 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 4 43 theme B 776:776 arg1 production 785:794 inhibin B (INHB) production 768:794 inhibin B (INHB) production in immature SCs isolated from 8-day-old rats 768:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 1 44 theme follicle-stimulating 97:116 arg1 FSH 127:129 FSH 127:129 FSH 127:129 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 1 44 theme follicle-stimulating 97:116 arg1 hormone 118:124 follicle-stimulating hormone 97:124 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 7 45 theme maturation 1165:1174 arg1 stage 1176:1180 a more advanced maturation stage 1149:1180 a more advanced maturation stage 1149:1180 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 2 46 theme FSH 346:348 arg1 structure 366:374 FSH oligosaccharide structure 346:374 FSH oligosaccharide structure 346:374 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 1 47 theme Sertoli 238:244 arg1 SCs 253:255 SCs 253:255 SCs 253:255 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 1 47 theme Sertoli 238:244 arg1 cells 246:250 Sertoli cells 238:250 Sertoli cells (SCs) 238:256 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 2 48 with structure 366:374 arg1 factors 409:415 gonadal factors 401:415 gonadal factors 401:415 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 6 49 gly acidic/sialylated 934:950 arg1 BA 976:977 BA 976:977 BA 976:977 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 49 gly acidic/sialylated 934:950 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 49 gly acidic/sialylated 934:950 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 11 50 theme production 1755:1764 arg1 regulation 1733:1742 the regulation 1729:1742 the regulation of inhibin production in the male gonad 1729:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 6 51 theme analogue 1001:1008 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 51 theme analogue 1001:1008 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 9 52 from effect 1394:1399 arg1 INHB 1404:1407 INHB 1404:1407 INHB 1404:1407 The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. 28855248 9 52 from effect 1394:1399 arg1 Pro-αC 1413:1418 Pro-αC 1413:1418 Pro-αC 1413:1418 The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. 28855248 3 53 theme human 533:537 arg1 rhFSH 544:548 rhFSH 544:548 rhFSH 544:548 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 53 theme human 533:537 arg1 FSH 539:541 Recombinant human FSH 521:541 Recombinant human FSH (rhFSH) glycosylation variants 521:572 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 10 54 theme gonadal 1620:1626 arg1 factors 1635:1641 gonadal growth factors 1620:1641 gonadal growth factors 1620:1641 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 2 55 from impact 336:341 arg1 regulation 424:433 the regulation 420:433 the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC 420:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 2 56 theme present 299:305 arg1 study 307:311 the present study 295:311 the present study 295:311 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 7 57 from stage 1176:1180 arg1 SCs 1142:1144 SCs 1142:1144 SCs 1142:1144 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 6 58 theme potent 1055:1060 arg1 stimulus 1062:1069 the most potent stimulus 1046:1069 INHB as well as the most potent stimulus for Pro-αC production 1030:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 10 59 theme hybrid-type 1469:1479 arg1 oligosaccharides 1481:1496 high mannose and hybrid-type oligosaccharides 1452:1496 oligosaccharides 1481:1496 Glycoforms bearing high mannose and hybrid-type oligosaccharides, HY, stimulated INHB and Pro-αC more effectively than those bearing complex oligosaccharides, CO, even in the presence of gonadal growth factors. 28855248 9 60 theme BA 1345:1346 arg1 preparation 1348:1358 The BA preparation 1341:1358 The BA preparation 1341:1358 The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. 28855248 8 61 theme factors 1332:1338 arg1 presence 1313:1320 the presence 1309:1320 the presence of growth factors 1309:1338 In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. 28855248 5 62 contain had 872:874 arg1 A 850:850 Activin A 842:850 Activin A 842:850 Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. 28855248 5 62 contain had 872:874 arg2 effect 890:895 a synergistic effect 876:895 a synergistic effect on FSH to stimulate Pro-αC 876:922 Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. 28855248 11 63 theme modulatory 1675:1684 arg1 effect 1686:1691 the modulatory effect 1671:1691 the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad 1671:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 5 64 theme Activin 842:848 arg1 A 850:850 Activin A 842:850 Activin A 842:850 Activin A stimulated INHB and had a synergistic effect on FSH to stimulate Pro-αC. 28855248 11 65 theme oligosaccharide 1700:1714 arg1 structure 1716:1724 FSH oligosaccharide structure 1696:1724 FSH oligosaccharide structure 1696:1724 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 1 66 from changes 227:233 arg1 SCs 253:255 SCs 253:255 SCs 253:255 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 1 66 from changes 227:233 arg1 cells 246:250 Sertoli cells 238:250 Sertoli cells (SCs) 238:256 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 3 67 theme sialylation 607:617 arg1 degree 619:624 their sialylation degree 601:624 their sialylation degree (AC and BA) 601:636 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 4 68 from effect 758:763 arg1 production 785:794 inhibin B (INHB) production 768:794 inhibin B (INHB) production in immature SCs isolated from 8-day-old rats 768:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 9 69 theme stimulatory 1382:1392 arg1 effect 1394:1399 a more marked stimulatory effect 1368:1399 a more marked stimulatory effect on INHB and Pro-αC than the AC 1368:1430 The BA preparation exerted a more marked stimulatory effect on INHB and Pro-αC than the AC. 28855248 6 70 theme charge 958:963 arg1 BA 976:977 BA 976:977 BA 976:977 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 70 theme charge 958:963 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 70 theme charge 958:963 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 0 71 theme Sertoli 50:56 arg1 production 71:80 Sertoli cell inhibin production 50:80 Sertoli cell inhibin production 50:80 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 1 72 theme important 191:199 arg1 changes 227:233 important structural and functional changes 191:233 important structural and functional changes in Sertoli cells (SCs) during sexual maturation 191:281 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 4 73 attach isolated 812:819 arg1 rats 836:839 8-day-old rats 826:839 8-day-old rats 826:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 4 73 attach isolated 812:819 arg2 SCs 808:810 immature SCs 799:810 immature SCs isolated from 8-day-old rats 799:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 6 74 theme acidic/sialylated 934:950 arg1 BA 976:977 BA 976:977 BA 976:977 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 74 theme acidic/sialylated 934:950 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 74 theme acidic/sialylated 934:950 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 2 75 theme different 482:490 arg1 stages 503:508 different maturation stages 482:508 different maturation stages of the SC 482:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 0 76 theme inhibin 63:69 arg1 production 71:80 Sertoli cell inhibin production 50:80 Sertoli cell inhibin production 50:80 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 2 77 theme production 468:477 arg1 regulation 424:433 the regulation 420:433 the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC 420:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 7 78 theme Native 1094:1099 arg1 rhFSH 1101:1105 Native rhFSH 1094:1105 Native rhFSH 1094:1105 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 6 79 theme charge 994:999 arg1 analogues 965:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues 925:973 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 6 79 theme charge 994:999 arg1 mix 1010:1012 the only charge analogue mix 985:1012 the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production 985:1091 The less acidic/sialylated rhFSH charge analogues, BA, were the only charge analogue mix that stimulated INHB as well as the most potent stimulus for Pro-αC production. 28855248 8 80 theme rhFSH 1234:1238 arg1 variants 1254:1261 all rhFSH glycosylation variants 1230:1261 all rhFSH glycosylation variants 1230:1261 In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. 28855248 2 81 theme oligosaccharide 350:364 arg1 structure 366:374 FSH oligosaccharide structure 346:374 FSH oligosaccharide structure 346:374 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 0 82 theme glycan 12:17 arg1 structure 19:27 FSH glycan structure 8:27 FSH glycan structure 8:27 Role of FSH glycan structure in the regulation of Sertoli cell inhibin production. 28855248 7 83 theme 20-day-old 1197:1206 arg1 rats 1208:1211 20-day-old rats 1197:1211 20-day-old rats 1197:1211 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 4 84 theme inhibin 710:716 arg1 Pro-αC 729:734 Pro-αC 729:734 Pro-αC 729:734 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 4 84 theme inhibin 710:716 arg1 α-subunit 718:726 inhibin α-subunit 710:726 inhibin α-subunit (Pro-αC) 710:735 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 2 85 theme gonadal 401:407 arg1 factors 409:415 gonadal factors 401:415 gonadal factors 401:415 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 2 86 theme dimeric 452:458 arg1 production 468:477 monomeric and dimeric inhibin production 438:477 monomeric and dimeric inhibin production 438:477 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 8 87 theme Pro-αC 1282:1287 arg1 production 1289:1298 Pro-αC production 1282:1298 Pro-αC production 1282:1298 In these cells, all rhFSH glycosylation variants increased INHB and Pro-αC production, even in the presence of growth factors. 28855248 2 88 theme interaction 384:394 arg1 impact 336:341 the impact 332:341 the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC 332:518 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 2 89 theme monomeric 438:446 arg1 production 468:477 monomeric and dimeric inhibin production 438:477 monomeric and dimeric inhibin production 438:477 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 4 90 theme inhibin 768:774 arg1 INHB 779:782 INHB 779:782 INHB 779:782 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 4 90 theme inhibin 768:774 arg1 B 776:776 inhibin B 768:776 inhibin B (INHB) production in immature SCs isolated from 8-day-old rats 768:839 Native rhFSH stimulated inhibin α-subunit (Pro-αC) but did not show any effect on inhibin B (INHB) production in immature SCs isolated from 8-day-old rats. 28855248 11 91 theme inhibin 1747:1753 arg1 production 1755:1764 inhibin production 1747:1764 inhibin production 1747:1764 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 28855248 1 92 from Variations 83:92 arg1 structure 161:169 structure 161:169 structure 161:169 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 1 92 from Variations 83:92 arg1 composition 145:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 follicle-stimulating hormone (FSH) carbohydrate composition 97:155 Variations in follicle-stimulating hormone (FSH) carbohydrate composition and structure are associated with important structural and functional changes in Sertoli cells (SCs) during sexual maturation. 28855248 7 93 theme advanced 1156:1163 arg1 stage 1176:1180 a more advanced maturation stage 1149:1180 a more advanced maturation stage 1149:1180 Native rhFSH stimulated both Pro-αC and INHB in SCs at a more advanced maturation stage, isolated from 20-day-old rats. 28855248 2 94 theme study 307:311 arg1 aim 288:290 The aim 284:290 The aim of the present study 284:311 The aim of the present study was to investigate the impact of FSH oligosaccharide structure and its interaction with gonadal factors on the regulation of monomeric and dimeric inhibin production at different maturation stages of the SC. 28855248 3 95 dep degree 619:624 arg1 BA 634:635 BA 634:635 BA 634:635 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 3 95 dep degree 619:624 arg1 AC 627:628 AC 627:628 AC 627:628 Recombinant human FSH (rhFSH) glycosylation variants were isolated according to their sialylation degree (AC and BA) and complexity of oligosaccharides (CO and HY). 28855248 11 96 theme male 1773:1776 arg1 gonad 1778:1782 the male gonad 1769:1782 the male gonad 1769:1782 These findings demonstrate the modulatory effect of FSH oligosaccharide structure on the regulation of inhibin production in the male gonad. 27151270 8 0 theme sites 1164:1168 arg1 sites 1164:1168 the 13 sites 1157:1168 the 13 sites 1157:1168 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 8 0 theme sites 1164:1168 arg1 two 1150:1152 two 1150:1152 two 1150:1152 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 9 1 theme plant 1314:1318 arg1 glycoproteins 1329:1341 plant vacuolar glycoproteins 1314:1341 plant vacuolar glycoproteins 1314:1341 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 3 2 gly microheterogeneity 435:452 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 9 3 gly glycoproteins 1329:1341 arg1 glycoproteins 1329:1341 plant vacuolar glycoproteins 1314:1341 plant vacuolar glycoproteins 1314:1341 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 2 4 from tree 306:309 arg1 Peroxidases 284:294 Peroxidases 284:294 Peroxidases from palm tree leaves 284:316 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 1 5 theme enzyme 253:258 arg1 activity 260:267 full enzyme activity 248:267 full enzyme activity 248:267 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 9 6 theme vacuolar 1320:1327 arg1 glycoproteins 1329:1341 plant vacuolar glycoproteins 1314:1341 plant vacuolar glycoproteins 1314:1341 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 4 7 gly glycopeptide 666:677 arg2 glycopeptide 666:677 glycopeptide fragmentation spectra 666:699 glycopeptide fragmentation spectra 666:699 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 1 8 theme Class 174:178 arg1 peroxidases 185:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 1 8 theme Class 174:178 arg1 enzymes 207:213 redox enzymes 201:213 redox enzymes that rely on N-glycosylation for full enzyme activity and stability 201:281 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 9 9 theme glycoproteins 1329:1341 arg1 paucimannosidic 1279:1293 paucimannosidic 1279:1293 paucimannosidic 1279:1293 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 9 9 theme glycoproteins 1329:1341 arg1 characteristic 1296:1309 characteristic 1296:1309 characteristic of plant vacuolar glycoproteins 1296:1341 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 3 10 gly glycosylation 417:429 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 9 11 theme complex 1206:1212 arg1 glycans 1225:1231 complex and hybrid glycans 1206:1231 glycans 1225:1231 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 2 12 theme palm 301:304 arg1 tree 306:309 palm tree 301:309 palm tree leaves 301:316 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 4 13 theme glycopeptide 666:677 arg1 spectra 693:699 glycopeptide fragmentation spectra 666:699 glycopeptide fragmentation spectra 666:699 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 8 14 used occupied 1110:1117 arg2 quantitation 1053:1064 Relative quantitation 1044:1064 Relative quantitation revealed 90% of each glycosylation site 1044:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 5 15 link N-linked 859:866 arg1 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 10 16 theme Further 1344:1350 arg1 studies 1352:1358 Further studies 1344:1358 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases 1344:1437 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases can benefit from the work outlined here. 27151270 5 17 theme windmill 885:892 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 0 18 theme Glycopeptide 120:131 arg1 Data 151:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 4 19 theme relative 725:732 arg1 glycoRQ 762:768 glycoRQ 762:768 glycoRQ 762:768 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 4 19 theme relative 725:732 arg1 quantitation 734:745 relative quantitation 725:745 relative quantitation of glycoforms 725:759 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 8 20 theme site 1101:1104 arg1 site 1101:1104 each glycosylation site 1082:1104 each glycosylation site 1082:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 8 20 theme site 1101:1104 arg1 % 1077:1077 90% 1075:1077 90% of each glycosylation site 1075:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 9 21 theme hybrid 1218:1223 arg1 glycans 1225:1231 complex and hybrid glycans 1206:1231 glycans 1225:1231 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 5 22 theme tree 899:902 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 3 23 theme palm 466:469 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 0 24 theme Plant 114:118 arg1 Data 151:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 6 25 theme Glycan 916:921 arg1 microheterogeneity 923:940 Glycan microheterogeneity 916:940 Glycan microheterogeneity 916:940 Glycan microheterogeneity was observed at each site. 27151270 8 26 theme glycosylation 1087:1099 arg1 site 1101:1104 each glycosylation site 1082:1104 each glycosylation site 1082:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 8 27 theme Relative 1044:1051 arg1 quantitation 1053:1064 Relative quantitation 1044:1064 Relative quantitation revealed 90% of each glycosylation site 1044:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 4 28 theme plant 605:609 arg1 pGlycoFilter 634:645 pGlycoFilter 634:645 pGlycoFilter 634:645 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 4 28 theme plant 605:609 arg1 identification 618:631 plant glycan identification 605:631 plant glycan identification 605:631 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 3 29 theme tree 471:474 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 0 30 theme N-Glycosylation 14:28 arg1 Characterization 30:45 Site-Specific N-Glycosylation Characterization 0:45 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 0:155 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 4 31 dep tools 577:581 arg1 aid 601:603 aid 601:603 to aid plant glycan identification, pGlycoFilter 598:645 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 5 32 theme Mass 771:774 arg1 analysis 789:796 Mass spectrometry analysis 771:796 Mass spectrometry analysis 771:796 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 33 from sites 850:854 arg1 profile 832:838 a detailed glycosylation profile 807:838 a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 807:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 33 from sites 850:854 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 4 34 theme spectra 693:699 arg1 annotation 652:661 annotation 652:661 annotation of glycopeptide fragmentation spectra 652:699 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 4 34 theme spectra 693:699 arg1 gPSMvalidator 702:714 gPSMvalidator 702:714 gPSMvalidator 702:714 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 0 35 theme Site-Specific 0:12 arg1 Characterization 30:45 Site-Specific N-Glycosylation Characterization 0:45 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 0:155 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 10 36 from relationships 1404:1416 arg1 peroxidases 1427:1437 plant peroxidases 1421:1437 plant peroxidases 1421:1437 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases can benefit from the work outlined here. 27151270 5 37 gly glycosylation 868:880 arg2 sites 850:854 the 13 sites 843:854 the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 843:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 37 gly glycosylation 868:880 arg2 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 37 gly glycosylation 868:880 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 4 38 theme glycoforms 750:759 arg1 glycoRQ 762:768 glycoRQ 762:768 glycoRQ 762:768 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 4 38 theme glycoforms 750:759 arg1 quantitation 734:745 relative quantitation 725:745 relative quantitation of glycoforms 725:759 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 3 39 theme windmill 457:464 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 0 40 theme Spectrometry 138:149 arg1 Data 151:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 7 41 theme Site 969:972 arg1 Asn211 974:979 Site Asn211 969:979 Site Asn211 969:979 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 7 41 theme Site 969:972 arg1 heterogeneous 994:1006 heterogeneous 994:1006 heterogeneous 994:1006 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 5 42 theme glycosylation 818:830 arg1 profile 832:838 a detailed glycosylation profile 807:838 a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 807:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 1 43 theme redox 201:205 arg1 peroxidases 185:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 1 43 theme redox 201:205 arg1 enzymes 207:213 redox enzymes 201:213 redox enzymes that rely on N-glycosylation for full enzyme activity and stability 201:281 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 5 44 from peroxidase 904:913 arg1 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 44 from peroxidase 904:913 arg1 sites 850:854 the 13 sites 843:854 the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 843:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 0 45 theme Mass 133:136 arg1 Data 151:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Plant Glycopeptide Mass Spectrometry Data 114:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 9 46 theme glycans 1266:1272 arg1 paucimannosidic 1279:1293 paucimannosidic 1279:1293 paucimannosidic 1279:1293 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 9 46 theme glycans 1266:1272 arg1 majority 1254:1261 the majority 1250:1261 the majority of glycans 1250:1272 Although complex and hybrid glycans were identified, the majority of glycans were paucimannosidic, characteristic of plant vacuolar glycoproteins. 27151270 2 47 theme plant 354:358 arg1 peroxidases 360:370 the most stable and active plant peroxidases 327:370 the most stable and active plant peroxidases characterized to date 327:392 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 7 48 contain contained 1012:1020 arg1 Asn211 974:979 Site Asn211 969:979 Site Asn211 969:979 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 7 48 contain contained 1012:1020 arg1 heterogeneous 994:1006 heterogeneous 994:1006 heterogeneous 994:1006 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 7 48 contain contained 1012:1020 arg2 glycans 1035:1041 30 different glycans 1022:1041 30 different glycans 1022:1041 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 5 49 theme N-linked 859:866 arg1 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 0 50 theme Palm 59:62 arg1 Peroxidase 69:78 Windmill Palm Tree Peroxidase 50:78 Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data 50:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 6 51 located observed 946:953 arg2 microheterogeneity 923:940 Glycan microheterogeneity 916:940 Glycan microheterogeneity 916:940 Glycan microheterogeneity was observed at each site. 27151270 6 51 located observed 946:953 arg1 site 963:966 each site 958:966 each site 958:966 Glycan microheterogeneity was observed at each site. 27151270 4 52 theme fragmentation 679:691 arg1 spectra 693:699 glycopeptide fragmentation spectra 666:699 glycopeptide fragmentation spectra 666:699 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 5 53 theme glycosylation 868:880 arg1 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 53 theme glycosylation 868:880 arg1 sites 850:854 the 13 sites 843:854 the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 843:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 0 54 theme Windmill 50:57 arg1 Peroxidase 69:78 Windmill Palm Tree Peroxidase 50:78 Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data 50:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 0 55 theme Data 151:154 arg1 Analysis 102:109 Analysis 102:109 Analysis of Plant Glycopeptide Mass Spectrometry Data 102:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 5 56 from glycosylation 868:880 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 3 57 theme Trachycarpus 477:488 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 0 58 theme Peroxidase 69:78 arg1 Characterization 30:45 Site-Specific N-Glycosylation Characterization 0:45 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 0:155 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 5 59 theme spectrometry 776:787 arg1 analysis 789:796 Mass spectrometry analysis 771:796 Mass spectrometry analysis 771:796 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 10 60 theme glycan 1378:1383 arg1 relationships 1404:1416 the glycan structure-activity relationships 1374:1416 the glycan structure-activity relationships in plant peroxidases 1374:1437 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases can benefit from the work outlined here. 27151270 2 61 theme active 347:352 arg1 peroxidases 360:370 the most stable and active plant peroxidases 327:370 the most stable and active plant peroxidases characterized to date 327:392 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 0 62 theme Tree 64:67 arg1 Peroxidase 69:78 Windmill Palm Tree Peroxidase 50:78 Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data 50:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 4 63 theme novel 571:575 arg1 tools 577:581 novel tools 571:581 novel tools 571:581 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 2 64 dep tree 306:309 arg1 leaves 311:316 leaves 311:316 leaves 311:316 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 5 65 theme detailed 809:816 arg1 profile 832:838 a detailed glycosylation profile 807:838 a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 807:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 2 66 theme stable 336:341 arg1 peroxidases 360:370 the most stable and active plant peroxidases 327:370 the most stable and active plant peroxidases characterized to date 327:392 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 4 67 theme glycan 611:616 arg1 pGlycoFilter 634:645 pGlycoFilter 634:645 pGlycoFilter 634:645 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 4 67 theme glycan 611:616 arg1 identification 618:631 plant glycan identification 605:631 plant glycan identification 605:631 The workflow developed in this study includes novel tools, written in R, to aid plant glycan identification, pGlycoFilter, for annotation of glycopeptide fragmentation spectra, gPSMvalidator, and for relative quantitation of glycoforms, glycoRQ. 27151270 10 68 theme structure-activity 1385:1402 arg1 relationships 1404:1416 the glycan structure-activity relationships 1374:1416 the glycan structure-activity relationships in plant peroxidases 1374:1437 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases can benefit from the work outlined here. 27151270 7 69 theme different 1025:1033 arg1 glycans 1035:1041 30 different glycans 1022:1041 30 different glycans 1022:1041 Site Asn211 was the most heterogeneous and contained 30 different glycans. 27151270 2 70 theme most 331:334 arg1 peroxidases 360:370 the most stable and active plant peroxidases 327:370 the most stable and active plant peroxidases characterized to date 327:392 Peroxidases from palm tree leaves comprise the most stable and active plant peroxidases characterized to date. 27151270 3 71 theme peroxidase 500:509 arg1 glycosylation 417:429 site-specific glycosylation 403:429 site-specific glycosylation 403:429 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 3 71 theme peroxidase 500:509 arg1 microheterogeneity 435:452 microheterogeneity 435:452 microheterogeneity 435:452 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 0 72 theme Novel 86:90 arg1 Tools 92:96 Novel Tools 86:96 Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data 86:154 Site-Specific N-Glycosylation Characterization of Windmill Palm Tree Peroxidase Using Novel Tools for Analysis of Plant Glycopeptide Mass Spectrometry Data. 27151270 10 73 theme plant 1421:1425 arg1 peroxidases 1427:1437 plant peroxidases 1421:1437 plant peroxidases 1421:1437 Further studies pertaining to the glycan structure-activity relationships in plant peroxidases can benefit from the work outlined here. 27151270 3 74 theme fortunei 490:497 arg1 peroxidase 500:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 windmill palm tree (Trachycarpus fortunei) peroxidase 457:509 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 1 75 theme Plant 157:161 arg1 peroxidases 185:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 1 75 theme Plant 157:161 arg1 enzymes 207:213 redox enzymes 201:213 redox enzymes that rely on N-glycosylation for full enzyme activity and stability 201:281 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 5 76 theme palm 894:897 arg1 peroxidase 904:913 windmill palm tree peroxidase 885:913 windmill palm tree peroxidase 885:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 3 77 theme site-specific 403:415 arg1 glycosylation 417:429 site-specific glycosylation 403:429 site-specific glycosylation 403:429 Herein, site-specific glycosylation and microheterogeneity of windmill palm tree (Trachycarpus fortunei) peroxidase are reported. 27151270 5 78 gly glycosylation 818:830 arg2 sites 850:854 the 13 sites 843:854 the 13 sites of N-linked glycosylation on windmill palm tree peroxidase 843:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 5 78 gly glycosylation 818:830 arg2 glycosylation 868:880 N-linked glycosylation 859:880 N-linked glycosylation on windmill palm tree peroxidase 859:913 Mass spectrometry analysis provided a detailed glycosylation profile at the 13 sites of N-linked glycosylation on windmill palm tree peroxidase. 27151270 1 79 theme secretory 163:171 arg1 peroxidases 185:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases 157:195 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 1 79 theme secretory 163:171 arg1 enzymes 207:213 redox enzymes 201:213 redox enzymes that rely on N-glycosylation for full enzyme activity and stability 201:281 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 27151270 8 80 gly glycosylation 1087:1099 arg2 site 1101:1104 each glycosylation site 1082:1104 each glycosylation site 1082:1104 Relative quantitation revealed 90% of each glycosylation site was occupied by three or fewer glycans, and two of the 13 sites were partially unoccupied. 27151270 1 81 theme full 248:251 arg1 activity 260:267 full enzyme activity 248:267 full enzyme activity 248:267 Plant secretory (Class III) peroxidases are redox enzymes that rely on N-glycosylation for full enzyme activity and stability. 24549892 5 0 with protein 1097:1103 arg1 sites 1123:1127 16 predicted sites 1110:1127 16 predicted sites 1110:1127 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 1 1 theme glycosylation 294:306 arg1 lack 328:331 the lack 324:331 the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies 324:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 1 theme glycosylation 294:306 arg1 diversity 281:289 diversity 281:289 diversity 281:289 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 1 theme glycosylation 294:306 arg1 heterogeneity 263:275 heterogeneity 263:275 heterogeneity 263:275 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 7 2 gly glycoproteins 1699:1711 arg1 glycoproteins 1699:1711 complex relatively pure glycoproteins 1675:1711 complex relatively pure glycoproteins 1675:1711 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 6 3 theme retention 1270:1278 arg1 times 1280:1284 the retention times 1266:1284 the retention times of glycopeptides 1266:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 3 4 theme site-specific 773:785 arg1 mapping 803:809 site-specific N-glycosylation mapping 773:809 site-specific N-glycosylation mapping 773:809 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 6 5 theme glycoform 1566:1574 arg1 compositions 1452:1463 glycan compositions 1445:1463 glycan compositions 1445:1463 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 5 theme glycoform 1566:1574 arg1 sequences 1477:1485 peptide sequences 1469:1485 peptide sequences 1469:1485 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 5 theme glycoform 1566:1574 arg1 abundances 1547:1556 relative abundances 1538:1556 relative abundances 1538:1556 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 5 theme glycoform 1566:1574 arg1 occupancy 1524:1532 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 4 6 theme experimental 962:973 arg1 pipeline 975:982 an experimental pipeline 959:982 an experimental pipeline with parallel analysis of glyco- and deglycopeptides 959:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 3 7 theme dissociation 842:853 arg1 fragmentation 866:878 collision-induced dissociation (CID)-only fragmentation 824:878 collision-induced dissociation (CID)-only fragmentation 824:878 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 3 8 theme -only 860:864 arg1 fragmentation 866:878 collision-induced dissociation (CID)-only fragmentation 824:878 collision-induced dissociation (CID)-only fragmentation 824:878 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 1 9 gly glycopeptide 155:166 arg2 glycopeptide 155:166 Site-specific glycopeptide 141:166 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS 141:233 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 4 10 theme glyco- 1010:1015 arg1 analysis 998:1005 parallel analysis 989:1005 parallel analysis of glyco- and deglycopeptides 989:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 1 11 dep heterogeneity 263:275 arg1 the 259:261 the 259:261 the 259:261 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 6 12 theme non-glycolytic 1332:1345 arg1 peptide 1347:1353 the non-glycolytic peptide 1328:1353 the non-glycolytic peptide from which the glycopeptide was derived 1328:1393 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 13 theme complex 628:634 arg1 mixtures 636:643 complex mixtures 628:643 complex mixtures of glycopeptides 628:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 1 14 from diversity 281:289 arg1 proteins 311:318 proteins 311:318 proteins 311:318 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 15 theme Site-specific 141:153 arg1 glycopeptide 155:166 Site-specific glycopeptide 141:166 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS 141:233 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 4 16 theme deglycopeptides 1021:1035 arg1 analysis 998:1005 parallel analysis 989:1005 parallel analysis of glyco- and deglycopeptides 989:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 3 17 theme retention 901:909 arg1 times 911:915 chromatographic retention times 885:915 chromatographic retention times of glycopeptides 885:932 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 1 18 from heterogeneity 263:275 arg1 proteins 311:318 proteins 311:318 proteins 311:318 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 5 19 theme test 1045:1048 arg1 case 1050:1053 the test case 1041:1053 the test case 1041:1053 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 5 19 theme test 1045:1048 arg1 ABCA4 1064:1068 ABCA4 1064:1068 ABCA4 1064:1068 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 7 20 theme complex 1739:1745 arg1 mixtures 1760:1767 more complex glycoprotein mixtures 1734:1767 more complex glycoprotein mixtures 1734:1767 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 6 21 theme N-glycosylation 1503:1517 arg1 occupancy 1524:1532 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 22 with glycopeptides 527:539 arg1 errors 556:561 mass errors 551:561 mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 551:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 1 23 theme simultaneous 180:191 arg1 glycan 193:198 simultaneous glycan and peptide characterization 180:227 glycan 193:198 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 0 24 theme collision-induced 81:97 arg1 MS/MS 99:103 collision-induced MS/MS 81:103 collision-induced MS/MS 81:103 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 25 gly heterogeneity 263:275 arg1 glycosylation 294:306 glycosylation 294:306 glycosylation 294:306 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 25 gly heterogeneity 263:275 arg1 information 359:369 complete fragmentation information 336:369 complete fragmentation information for either peptides or glycans with current fragmentation technologies 336:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 4 26 gly deglycopeptides 1021:1035 arg2 deglycopeptides 1021:1035 deglycopeptides 1021:1035 deglycopeptides 1021:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 7 27 theme pure 1694:1697 arg1 glycoproteins 1699:1711 complex relatively pure glycoproteins 1675:1711 complex relatively pure glycoproteins 1675:1711 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 2 28 theme large 703:707 arg1 proteins 709:716 large proteins 703:716 large proteins 703:716 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 2 29 theme multiple 451:458 arg1 peptide 460:466 multiple peptide and glycan combinations 451:490 peptide 460:466 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 0 30 theme retention 125:133 arg1 time 135:138 chromatographic retention time 109:138 chromatographic retention time 109:138 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 31 theme fragmentation 415:427 arg1 technologies 429:440 current fragmentation technologies 407:440 current fragmentation technologies 407:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 5 32 theme integral 1079:1086 arg1 ABCA4 1064:1068 ABCA4 1064:1068 ABCA4 1064:1068 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 5 32 theme integral 1079:1086 arg1 protein 1097:1103 a large integral membrane protein 1071:1103 a large integral membrane protein with 16 predicted sites for N-glycosylation 1071:1147 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 0 33 theme Reliable 0:7 arg1 determination 9:21 Reliable determination 0:21 Reliable determination of site-specific in vivo protein N-glycosylation 0:70 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 6 34 theme CID 1170:1172 arg1 speed 1201:1205 high scan speed 1191:1205 high scan speed 1191:1205 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 34 theme CID 1170:1172 arg1 features 1174:1181 CID features 1170:1181 CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides 1170:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 34 theme CID 1170:1172 arg1 intensity 1216:1224 high intensity 1211:1224 high intensity 1211:1224 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 1 35 theme lack 328:331 arg1 difficult 238:246 difficult 238:246 difficult 238:246 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 6 36 dep occupancy 1524:1532 arg1 the 1499:1501 the 1499:1501 the 1499:1501 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 37 theme glycopeptides 527:539 arg1 mass 519:522 the same mass 510:522 the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 510:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 0 38 theme in 40:41 arg1 N-glycosylation 56:70 site-specific in vivo protein N-glycosylation 26:70 site-specific in vivo protein N-glycosylation 26:70 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 39 theme complete 336:343 arg1 information 359:369 complete fragmentation information 336:369 complete fragmentation information for either peptides or glycans with current fragmentation technologies 336:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 6 40 theme relative 1538:1545 arg1 abundances 1547:1556 relative abundances 1538:1556 relative abundances 1538:1556 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 41 gly glycopeptides 527:539 arg2 glycopeptides 527:539 glycopeptides 527:539 glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 527:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 42 from sites 1588:1592 arg1 compositions 1452:1463 glycan compositions 1445:1463 glycan compositions 1445:1463 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 42 from sites 1588:1592 arg1 sequences 1477:1485 peptide sequences 1469:1485 peptide sequences 1469:1485 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 42 from sites 1588:1592 arg1 abundances 1547:1556 relative abundances 1538:1556 relative abundances 1538:1556 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 42 from sites 1588:1592 arg1 occupancy 1524:1532 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 0 43 theme protein 48:54 arg1 N-glycosylation 56:70 site-specific in vivo protein N-glycosylation 26:70 site-specific in vivo protein N-glycosylation 26:70 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 44 theme information 359:369 arg1 lack 328:331 the lack 324:331 the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies 324:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 44 theme information 359:369 arg1 diversity 281:289 diversity 281:289 diversity 281:289 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 44 theme information 359:369 arg1 heterogeneity 263:275 heterogeneity 263:275 heterogeneity 263:275 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 0 45 dep in 40:41 arg1 vivo 43:46 vivo 43:46 vivo 43:46 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 2 46 from case 695:698 arg1 challenging 676:686 challenging 676:686 challenging 676:686 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 3 47 theme novel 736:740 arg1 strategy 742:749 a novel strategy 734:749 a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides 734:932 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 6 48 theme high 1211:1214 arg1 intensity 1216:1224 high intensity 1211:1224 high intensity 1211:1224 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 49 gly glycopeptides 1289:1301 arg2 glycopeptides 1289:1301 glycopeptides 1289:1301 glycopeptides 1289:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 50 theme high 1191:1194 arg1 speed 1201:1205 high scan speed 1191:1205 high scan speed 1191:1205 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 51 theme specific 1579:1586 arg1 sites 1588:1592 specific sites 1579:1592 specific sites for ABCA4 1579:1602 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 52 theme glycopeptides 648:660 arg1 mixtures 636:643 complex mixtures 628:643 complex mixtures of glycopeptides 628:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 1 53 from lack 328:331 arg1 proteins 311:318 proteins 311:318 proteins 311:318 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 54 theme peptide 204:210 arg1 characterization 212:227 simultaneous glycan and peptide characterization 180:227 characterization 212:227 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 3 55 theme N-glycosylation 787:801 arg1 mapping 803:809 site-specific N-glycosylation mapping 773:809 site-specific N-glycosylation mapping 773:809 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 6 56 theme peptide 1469:1475 arg1 sequences 1477:1485 peptide sequences 1469:1485 peptide sequences 1469:1485 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 57 theme ions 1238:1241 arg1 speed 1201:1205 high scan speed 1191:1205 high scan speed 1191:1205 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 57 theme ions 1238:1241 arg1 intensity 1216:1224 high intensity 1211:1224 high intensity 1211:1224 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 58 attach derived 1387:1393 arg1 peptide 1347:1353 the non-glycolytic peptide 1328:1353 the non-glycolytic peptide from which the glycopeptide was derived 1328:1393 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 58 attach derived 1387:1393 arg2 glycopeptide 1370:1381 the glycopeptide 1366:1381 the glycopeptide 1366:1381 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 3 59 theme collision-induced 824:840 arg1 dissociation 842:853 collision-induced dissociation 824:853 collision-induced dissociation (CID)-only fragmentation 824:878 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 3 59 theme collision-induced 824:840 arg1 CID 856:858 CID 856:858 CID 856:858 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 2 60 gly glycopeptides 648:660 arg2 glycopeptides 648:660 glycopeptides 648:660 glycopeptides 648:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 3 61 gly glycopeptides 920:932 arg2 glycopeptides 920:932 glycopeptides 920:932 glycopeptides 920:932 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 6 62 gly glycopeptide 1370:1381 arg2 glycopeptide 1370:1381 the glycopeptide 1366:1381 the glycopeptide 1366:1381 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 7 63 theme glycoprotein 1747:1758 arg1 mixtures 1760:1767 more complex glycoprotein mixtures 1734:1767 more complex glycoprotein mixtures 1734:1767 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 2 64 theme mixtures 636:643 arg1 analysis 616:623 analysis 616:623 analysis of complex mixtures of glycopeptides 616:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 2 64 theme mixtures 636:643 arg1 ambiguity 602:610 ambiguity 602:610 ambiguity 602:610 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 65 theme complete 1418:1425 arg1 information 1427:1437 virtually complete information 1408:1437 virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4 1408:1602 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 5 66 theme membrane 1088:1095 arg1 ABCA4 1064:1068 ABCA4 1064:1068 ABCA4 1064:1068 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 5 66 theme membrane 1088:1095 arg1 protein 1097:1103 a large integral membrane protein 1071:1103 a large integral membrane protein with 16 predicted sites for N-glycosylation 1071:1147 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 4 67 theme parallel 989:996 arg1 analysis 998:1005 parallel analysis 989:1005 parallel analysis of glyco- and deglycopeptides 989:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 6 68 theme site 1519:1522 arg1 occupancy 1524:1532 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 4 69 with pipeline 975:982 arg1 analysis 998:1005 parallel analysis 989:1005 parallel analysis of glyco- and deglycopeptides 989:1035 This approach leverages an experimental pipeline with parallel analysis of glyco- and deglycopeptides. 24549892 2 70 theme mass 551:554 arg1 errors 556:561 mass errors 551:561 mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 551:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 71 theme glycopeptides 1289:1301 arg1 times 1280:1284 the retention times 1266:1284 the retention times of glycopeptides 1266:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 3 72 theme chromatographic 885:899 arg1 times 911:915 chromatographic retention times 885:915 chromatographic retention times of glycopeptides 885:932 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 1 73 with glycans 394:400 arg1 technologies 429:440 current fragmentation technologies 407:440 current fragmentation technologies 407:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 0 74 theme chromatographic 109:123 arg1 time 135:138 chromatographic retention time 109:138 chromatographic retention time 109:138 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 75 with peptides 382:389 arg1 technologies 429:440 current fragmentation technologies 407:440 current fragmentation technologies 407:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 2 76 theme proteins 709:716 arg1 case 695:698 the case 691:698 the case of large proteins 691:716 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 3 77 theme glycopeptides 920:932 arg1 times 911:915 chromatographic retention times 885:915 chromatographic retention times of glycopeptides 885:932 Here we report a novel strategy to reliably determine site-specific N-glycosylation mapping by combining collision-induced dissociation (CID)-only fragmentation with chromatographic retention times of glycopeptides. 24549892 1 78 theme current 407:413 arg1 technologies 429:440 current fragmentation technologies 407:440 current fragmentation technologies 407:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 5 79 theme large 1073:1077 arg1 ABCA4 1064:1068 ABCA4 1064:1068 ABCA4 1064:1068 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 5 79 theme large 1073:1077 arg1 protein 1097:1103 a large integral membrane protein 1071:1103 a large integral membrane protein with 16 predicted sites for N-glycosylation 1071:1147 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 2 80 theme less 563:566 arg1 errors 556:561 mass errors 551:561 mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 551:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 81 theme features 1174:1181 arg1 advantage 1157:1165 advantage 1157:1165 advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides 1157:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 0 82 theme site-specific 26:38 arg1 N-glycosylation 56:70 site-specific in vivo protein N-glycosylation 26:70 site-specific in vivo protein N-glycosylation 26:70 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 2 83 from challenging 676:686 arg1 case 695:698 the case 691:698 the case of large proteins 691:716 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 84 gly occupancy 1524:1532 arg2 site 1519:1522 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 84 gly occupancy 1524:1532 arg2 glycoform 1566:1574 each glycoform 1561:1574 each glycoform 1561:1574 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 84 gly occupancy 1524:1532 arg1 glycoform 1566:1574 each glycoform 1561:1574 each glycoform 1561:1574 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 85 theme glycan 472:477 arg1 combinations 479:490 multiple peptide and glycan combinations 451:490 combinations 479:490 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 7 86 theme complex 1675:1681 arg1 glycoproteins 1699:1711 complex relatively pure glycoproteins 1675:1711 complex relatively pure glycoproteins 1675:1711 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 5 87 theme predicted 1113:1121 arg1 sites 1123:1127 16 predicted sites 1110:1127 16 predicted sites 1110:1127 As the test case we chose ABCA4, a large integral membrane protein with 16 predicted sites for N-glycosylation. 24549892 0 88 theme N-glycosylation 56:70 arg1 determination 9:21 Reliable determination 0:21 Reliable determination of site-specific in vivo protein N-glycosylation 0:70 Reliable determination of site-specific in vivo protein N-glycosylation based on collision-induced MS/MS and chromatographic retention time. 24549892 1 89 theme fragmentation 345:357 arg1 information 359:369 complete fragmentation information 336:369 complete fragmentation information for either peptides or glycans with current fragmentation technologies 336:440 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 6 90 theme fragment 1229:1236 arg1 ions 1238:1241 fragment ions 1229:1241 fragment ions together combined with the retention times of glycopeptides 1229:1301 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 2 91 theme same 514:517 arg1 mass 519:522 the same mass 510:522 the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides 510:660 Indeed, multiple peptide and glycan combinations can readily match the same mass of glycopeptides even with mass errors less than 5 ppm providing considerably ambiguity and analysis of complex mixtures of glycopeptides becomes quite challenging in the case of large proteins. 24549892 6 92 dep speed 1201:1205 arg1 scan 1196:1199 scan 1196:1199 scan 1196:1199 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 93 gly N-glycosylation 1503:1517 arg2 site 1519:1522 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 6 93 gly N-glycosylation 1503:1517 arg2 occupancy 1524:1532 N-glycosylation site occupancy 1503:1532 N-glycosylation site occupancy 1503:1532 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 7 94 gly glycoprotein 1747:1758 arg1 glycoprotein 1747:1758 more complex glycoprotein mixtures 1734:1767 more complex glycoprotein mixtures 1734:1767 The challenges provided by this example provide guidance in analyzing complex relatively pure glycoproteins and potentially even more complex glycoprotein mixtures. 24549892 6 95 theme glycan 1445:1450 arg1 compositions 1452:1463 glycan compositions 1445:1463 glycan compositions 1445:1463 Taking advantage of CID features such as high scan speed and high intensity of fragment ions together combined with the retention times of glycopeptides to conclusively identify the non-glycolytic peptide from which the glycopeptide was derived, we obtained virtually complete information about glycan compositions and peptide sequences, as well as the N-glycosylation site occupancy and relative abundances of each glycoform at specific sites for ABCA4. 24549892 1 96 theme heterogeneity 263:275 arg1 difficult 238:246 difficult 238:246 difficult 238:246 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 24549892 1 97 theme diversity 281:289 arg1 difficult 238:246 difficult 238:246 difficult 238:246 Site-specific glycopeptide mapping for simultaneous glycan and peptide characterization by MS is difficult because of the heterogeneity and diversity of glycosylation in proteins and the lack of complete fragmentation information for either peptides or glycans with current fragmentation technologies. 25551295 9 0 theme level 1735:1739 arg1 presence 1710:1717 the presence 1706:1717 the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming 1706:1832 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 8 1 theme GlcNAc 1468:1473 arg1 residue 1475:1481 a single [(13)C,(15)N]GlcNAc residue 1446:1481 a single [(13)C,(15)N]GlcNAc residue on each N-glycan 1446:1498 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 0 2 theme immunoglobulin 150:163 arg1 G1 165:166 immunoglobulin G1 150:166 immunoglobulin G1 150:166 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 9 3 dep complex-type 1870:1881 arg1 biantennary 1884:1894 biantennary 1884:1894 biantennary 1884:1894 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 9 4 theme interaction 1744:1754 arg1 level 1735:1739 an increased level 1722:1739 an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming 1722:1832 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 7 5 with Fc 1180:1181 arg1 Gnt1 1234:1237 Gnt1 1234:1237 Gnt1 1234:1237 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 5 with Fc 1180:1181 arg1 Gnt2 1243:1246 Gnt2 1243:1246 Gnt2 1243:1246 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 5 with Fc 1180:1181 arg1 GlcNAc 1270:1275 UDP-[(13)C,(15)N]GlcNAc 1253:1275 UDP-[(13)C,(15)N]GlcNAc 1253:1275 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 5 with Fc 1180:1181 arg1 glycosyltransferases 1212:1231 recombinantly expressed glycosyltransferases 1188:1231 recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) 1188:1247 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 6 theme complete 1289:1296 arg1 conversion 1308:1317 complete glycoform conversion 1289:1317 complete glycoform conversion 1289:1317 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 8 7 contain containing 1435:1444 arg2 residue 1475:1481 a single [(13)C,(15)N]GlcNAc residue 1446:1481 a single [(13)C,(15)N]GlcNAc residue on each N-glycan 1446:1498 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 8 7 contain containing 1435:1444 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 5 8 theme immunoglobulin 885:898 arg1 fragment 903:910 the immunoglobulin G1 fragment 881:910 the immunoglobulin G1 fragment crystallizable (Fc) 881:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 10 9 theme N-glycan 1951:1958 arg1 maturation 1960:1969 Fc N-glycan maturation 1948:1969 Fc N-glycan maturation 1948:1969 Thus, conformational restriction increases as Fc N-glycan maturation proceeds. 25551295 1 10 theme Asparagine-linked 169:185 arg1 N 188:188 N 188:188 N 188:188 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 10 theme Asparagine-linked 169:185 arg1 modification 236:247 a common eukaryotic protein modification 208:247 a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues 208:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 10 theme Asparagine-linked 169:185 arg1 glycosylation 191:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 3 11 theme nuclear 643:649 arg1 resonance 660:668 nuclear magnetic resonance 643:668 nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms 643:767 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 9 12 dep N-glycan 1768:1775 arg1 the 1764:1766 the 1764:1766 the 1764:1766 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 8 13 from residue 1475:1481 arg1 N-glycan 1491:1498 each N-glycan 1486:1498 each N-glycan 1486:1498 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 7 14 theme C 1262:1262 arg1 GlcNAc 1270:1275 UDP-[(13)C,(15)N]GlcNAc 1253:1275 UDP-[(13)C,(15)N]GlcNAc 1253:1275 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 8 15 dep single 1448:1453 arg1 C 1460:1460 (13)C 1456:1460 (13)C 1456:1460 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 6 16 theme UDP-[ 1046:1050 arg1 C 1055:1055 UDP-[(13)C, 1046:1056 C 1055:1055 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 6 16 theme UDP-[ 1046:1050 arg1 15 1058:1059 15 1058:1059 15 1058:1059 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 11 17 theme complex-type 2072:2083 arg1 N-glycan 2085:2092 a complex-type N-glycan 2070:2092 a complex-type N-glycan 2070:2092 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 8 18 theme heteronuclear 1367:1379 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 1 19 theme intramolecular 311:324 arg1 interactions 326:337 intramolecular interactions 311:337 intramolecular interactions between N-glycan and polypeptide residues 311:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 9 20 theme N-glycan 1668:1675 arg1 pathway 1688:1694 the N-glycan remodeling pathway 1664:1694 the N-glycan remodeling pathway 1664:1694 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 9 21 theme homogeneous 1607:1617 arg1 glycoforms 1619:1628 homogeneous glycoforms 1607:1628 homogeneous glycoforms 1607:1628 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 8 22 theme coherence 1396:1404 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 8 23 theme Gnt1 1421:1424 arg1 product 1426:1432 the Gnt1 product 1417:1432 the Gnt1 product 1417:1432 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 0 24 theme C 109:109 arg1 labeling 138:145 [(13)C,(15)N]-N-acetylglucosamine labeling 104:145 [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1 104:166 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 0 24 theme C 109:109 arg1 15 112:113 15 112:113 15 112:113 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 5 25 theme N 1013:1013 arg1 C 1007:1007 [(13)C 1002:1007 [(13)C 1002:1007 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 5 25 theme N 1013:1013 arg1 GlcNAc 1037:1042 GlcNAc 1037:1042 GlcNAc 1037:1042 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 5 25 theme N 1013:1013 arg1 -N-acetylglucosamine 1015:1034 (15)N]-N-acetylglucosamine 1009:1034 (15)N]-N-acetylglucosamine (GlcNAc) 1009:1043 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 0 26 theme Intramolecular 0:13 arg1 interactions 36:47 Intramolecular N-glycan/polypeptide interactions 0:47 Intramolecular N-glycan/polypeptide interactions 0:47 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 1 27 theme protein 262:268 arg1 folding 270:276 protein folding 262:276 protein folding 262:276 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 11 28 theme fundamental 2004:2014 arg1 reactions 2016:2024 fundamental reactions 2004:2024 fundamental reactions 2004:2024 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 5 29 theme fragment 903:910 arg1 glycoforms 867:876 immature glycoforms 858:876 immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) 858:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 6 30 theme C 1055:1055 arg1 GlcNAc 1063:1068 UDP-[(13)C,(15)N]GlcNAc 1046:1068 UDP-[(13)C,(15)N]GlcNAc 1046:1068 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 9 31 theme remodeling 1677:1686 arg1 pathway 1688:1694 the N-glycan remodeling pathway 1664:1694 the N-glycan remodeling pathway 1664:1694 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 5 32 theme immature 858:865 arg1 glycoforms 867:876 immature glycoforms 858:876 immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) 858:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 11 33 gly glycoprotein 2052:2063 arg1 glycoprotein 2052:2063 every glycoprotein 2046:2063 every glycoprotein with a complex-type N-glycan 2046:2092 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 3 34 theme glycan 701:706 arg1 remodeling 708:717 enzymatic glycan remodeling 691:717 enzymatic glycan remodeling 691:717 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 0 35 theme N-glycan 70:77 arg1 steps 90:94 multiple N-glycan remodeling steps 61:94 multiple N-glycan remodeling steps 61:94 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 10 36 theme conformational 1908:1921 arg1 restriction 1923:1933 conformational restriction 1908:1933 conformational restriction 1908:1933 Thus, conformational restriction increases as Fc N-glycan maturation proceeds. 25551295 3 37 theme resonance 660:668 arg1 techniques 670:679 nuclear magnetic resonance techniques 643:679 nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms 643:767 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 11 38 with glycoprotein 2052:2063 arg1 N-glycan 2085:2092 a complex-type N-glycan 2070:2092 a complex-type N-glycan 2070:2092 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 11 39 theme broad 2153:2157 arg1 range 2159:2163 a broad range 2151:2163 a broad range of glycoprotein studies 2151:2187 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 5 40 dep crystallizable 912:925 arg1 Fc 928:929 Fc 928:929 Fc 928:929 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 1 41 theme common 210:215 arg1 modification 236:247 a common eukaryotic protein modification 208:247 a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues 208:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 41 theme common 210:215 arg1 glycosylation 191:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 9 42 theme increased 1725:1733 arg1 level 1735:1739 an increased level 1722:1739 an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming 1722:1832 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 0 43 theme G1 165:166 arg1 labeling 138:145 [(13)C,(15)N]-N-acetylglucosamine labeling 104:145 [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1 104:166 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 0 43 theme G1 165:166 arg1 15 112:113 15 112:113 15 112:113 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 1 44 theme protein 228:234 arg1 modification 236:247 a common eukaryotic protein modification 208:247 a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues 208:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 44 theme protein 228:234 arg1 glycosylation 191:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 11 45 theme studies 2181:2187 arg1 range 2159:2163 a broad range 2151:2163 a broad range of glycoprotein studies 2151:2187 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 7 46 theme mass 1332:1335 arg1 spectrometry 1337:1348 mass spectrometry 1332:1348 mass spectrometry 1332:1348 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 6 47 theme C 1132:1132 arg1 glucose 1134:1140 [(13)C]glucose 1127:1140 [(13)C]glucose 1127:1140 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 6 47 theme C 1132:1132 arg1 13 1129:1130 13 1129:1130 13 1129:1130 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 7 48 theme expressed 1202:1210 arg1 Gnt2 1243:1246 Gnt2 1243:1246 Gnt2 1243:1246 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 48 theme expressed 1202:1210 arg1 Gnt1 1234:1237 Gnt1 1234:1237 Gnt1 1234:1237 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 48 theme expressed 1202:1210 arg1 glycosyltransferases 1212:1231 recombinantly expressed glycosyltransferases 1188:1231 recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) 1188:1247 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 49 theme glycoform 1298:1306 arg1 conversion 1308:1317 complete glycoform conversion 1289:1317 complete glycoform conversion 1289:1317 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 9 50 theme glycoforms 1619:1628 arg1 spectra 1596:1602 Similar spectra 1588:1602 Similar spectra 1588:1602 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 8 51 theme polypeptide 1566:1576 arg1 residues 1578:1585 polypeptide residues 1566:1585 polypeptide residues 1566:1585 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 5 52 theme crystallizable 912:925 arg1 fragment 903:910 the immunoglobulin G1 fragment 881:910 the immunoglobulin G1 fragment crystallizable (Fc) 881:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 0 53 theme N 115:115 arg1 labeling 138:145 [(13)C,(15)N]-N-acetylglucosamine labeling 104:145 [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1 104:166 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 0 53 theme N 115:115 arg1 15 112:113 15 112:113 15 112:113 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 8 54 dep C 1460:1460 arg1 N 1466:1466 (15)N 1462:1466 (15)N 1462:1466 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 9 55 from step 1801:1804 arg1 N-glycan 1768:1775 N-glycan 1768:1775 N-glycan 1768:1775 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 9 55 from step 1801:1804 arg1 polypeptide 1781:1791 polypeptide 1781:1791 polypeptide 1781:1791 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 0 56 theme -N-acetylglucosamine 117:136 arg1 labeling 138:145 [(13)C,(15)N]-N-acetylglucosamine labeling 104:145 [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1 104:166 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 0 56 theme -N-acetylglucosamine 117:136 arg1 15 112:113 15 112:113 15 112:113 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 8 57 theme Two-dimensional 1351:1365 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 9 58 theme mannose 1817:1823 arg1 trimming 1825:1832 mannose trimming 1817:1832 mannose trimming 1817:1832 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 4 59 theme applicable 786:795 arg1 methods 797:803 widely applicable methods 779:803 widely applicable methods 779:803 However, widely applicable methods do not yet exist. 25551295 3 60 theme techniques 670:679 arg1 combination 628:638 a combination 626:638 a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms 626:767 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 9 61 theme Similar 1588:1594 arg1 spectra 1596:1602 Similar spectra 1588:1602 Similar spectra 1588:1602 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 8 62 theme single-quantum 1381:1394 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 3 63 theme magnetic 651:658 arg1 resonance 660:668 nuclear magnetic resonance 643:668 nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms 643:767 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 5 64 theme homogeneous 951:961 arg1 state 963:967 a homogeneous state 949:967 a homogeneous state 949:967 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 5 65 theme technological 839:851 arg1 gap 853:855 this technological gap 834:855 this technological gap 834:855 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 2 66 theme glycoprotein 503:514 arg1 properties 485:494 inherent properties 476:494 inherent properties 476:494 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 2 66 theme glycoprotein 503:514 arg1 conformational 534:547 conformational 534:547 conformational 534:547 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 2 66 theme glycoprotein 503:514 arg1 heterogeneity 567:579 compositional heterogeneity 553:579 compositional heterogeneity 553:579 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 8 67 with interactions 1548:1559 arg1 residues 1578:1585 polypeptide residues 1566:1585 polypeptide residues 1566:1585 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 1 68 theme N-glycan 347:354 arg1 residues 372:379 N-glycan and polypeptide residues 347:379 N-glycan and polypeptide residues 347:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 2 69 theme compositional 553:565 arg1 heterogeneity 567:579 compositional heterogeneity 553:579 compositional heterogeneity 553:579 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 8 70 theme product 1426:1432 arg1 spectra 1406:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra 1351:1412 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 1 71 theme polypeptide 360:370 arg1 residues 372:379 N-glycan and polypeptide residues 347:379 N-glycan and polypeptide residues 347:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 6 72 theme N 1061:1061 arg1 GlcNAc 1063:1068 UDP-[(13)C,(15)N]GlcNAc 1046:1068 UDP-[(13)C,(15)N]GlcNAc 1046:1068 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 9 73 theme different 1641:1649 arg1 points 1651:1656 different points 1641:1656 different points along the N-glycan remodeling pathway 1641:1694 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 1 74 link Asparagine-linked 169:185 arg1 N 188:188 N 188:188 N 188:188 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 74 link Asparagine-linked 169:185 arg1 modification 236:247 a common eukaryotic protein modification 208:247 a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues 208:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 74 link Asparagine-linked 169:185 arg1 glycosylation 191:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 9 75 theme complex-type 1870:1881 arg1 form 1896:1899 a complex-type, biantennary form 1868:1899 a complex-type, biantennary form 1868:1899 Similar spectra of homogeneous glycoforms, halted at different points along the N-glycan remodeling pathway, revealed the presence of an increased level of interaction between the N-glycan and polypeptide at each step, including mannose trimming, as the N-glycan was converted to a complex-type, biantennary form. 25551295 0 76 theme N-glycan/polypeptide 15:34 arg1 interactions 36:47 Intramolecular N-glycan/polypeptide interactions 0:47 Intramolecular N-glycan/polypeptide interactions 0:47 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 8 77 theme single 1448:1453 arg1 residue 1475:1481 a single [(13)C,(15)N]GlcNAc residue 1446:1481 a single [(13)C,(15)N]GlcNAc residue on each N-glycan 1446:1498 Two-dimensional heteronuclear single-quantum coherence spectra of the Gnt1 product, containing a single [(13)C,(15)N]GlcNAc residue on each N-glycan, showed that the N-glycan is stabilized through interactions with polypeptide residues. 25551295 5 78 theme G1 900:901 arg1 fragment 903:910 the immunoglobulin G1 fragment 881:910 the immunoglobulin G1 fragment crystallizable (Fc) 881:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 2 79 gly glycoprotein 503:514 arg1 glycoprotein 503:514 the glycoprotein 499:514 the glycoprotein 499:514 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 2 80 theme N-glycan 451:458 arg1 relationship 430:441 the structure-activity relationship 407:441 the structure-activity relationship of each N-glycan 407:458 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 0 81 theme multiple 61:68 arg1 steps 90:94 multiple N-glycan remodeling steps 61:94 multiple N-glycan remodeling steps 61:94 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 11 82 theme glycoprotein 2052:2063 arg1 synthesis 2033:2041 the synthesis 2029:2041 the synthesis of every glycoprotein with a complex-type N-glycan 2029:2092 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 3 83 theme enzymatic 691:699 arg1 remodeling 708:717 enzymatic glycan remodeling 691:717 enzymatic glycan remodeling 691:717 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 2 84 theme structure-activity 411:428 arg1 relationship 430:441 the structure-activity relationship 407:441 the structure-activity relationship of each N-glycan 407:458 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 0 85 theme remodeling 79:88 arg1 steps 90:94 multiple N-glycan remodeling steps 61:94 multiple N-glycan remodeling steps 61:94 Intramolecular N-glycan/polypeptide interactions observed at multiple N-glycan remodeling steps through [(13)C,(15)N]-N-acetylglucosamine labeling of immunoglobulin G1. 25551295 11 86 gly glycoprotein 2168:2179 arg1 glycoprotein 2168:2179 glycoprotein studies 2168:2187 glycoprotein studies 2168:2187 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 6 87 theme one-pot 1105:1111 arg1 reaction 1113:1120 a one-pot reaction 1103:1120 a one-pot reaction 1103:1120 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 2 88 theme inherent 476:483 arg1 properties 485:494 inherent properties 476:494 inherent properties 476:494 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 2 88 theme inherent 476:483 arg1 conformational 534:547 conformational 534:547 conformational 534:547 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 2 88 theme inherent 476:483 arg1 heterogeneity 567:579 compositional heterogeneity 553:579 compositional heterogeneity 553:579 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 3 89 theme homogeneous 746:756 arg1 glycoforms 758:767 homogeneous glycoforms 746:767 homogeneous glycoforms 746:767 These limitations can be addressed by using a combination of nuclear magnetic resonance techniques following enzymatic glycan remodeling to simultaneously generate homogeneous glycoforms. 25551295 6 90 theme N-amido 1151:1157 arg1 glutamine 1159:1167 [(15)N-amido]glutamine 1146:1167 [(15)N-amido]glutamine 1146:1167 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 6 90 theme N-amido 1151:1157 arg1 15 1148:1149 15 1148:1149 15 1148:1149 UDP-[(13)C,(15)N]GlcNAc was synthesized enzymatically in a one-pot reaction from [(13)C]glucose and [(15)N-amido]glutamine. 25551295 11 91 theme glycoprotein 2168:2179 arg1 studies 2181:2187 glycoprotein studies 2168:2187 glycoprotein studies 2168:2187 Gnt1 and Gnt2 catalyze fundamental reactions in the synthesis of every glycoprotein with a complex-type N-glycan; thus, the strategies presented herein can be applied to a broad range of glycoprotein studies. 25551295 7 92 dep glycosyltransferases 1212:1231 arg1 Gnt2 1243:1246 Gnt2 1243:1246 Gnt2 1243:1246 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 92 dep glycosyltransferases 1212:1231 arg1 Gnt1 1234:1237 Gnt1 1234:1237 Gnt1 1234:1237 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 7 92 dep glycosyltransferases 1212:1231 arg1 glycosyltransferases 1212:1231 recombinantly expressed glycosyltransferases 1188:1231 recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) 1188:1247 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 2 93 dep Attempts 382:389 arg1 hindered 464:471 hindered 464:471 are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity 460:579 Attempts to characterize the structure-activity relationship of each N-glycan are hindered by inherent properties of the glycoprotein, including glycan conformational and compositional heterogeneity. 25551295 10 94 theme Fc 1948:1949 arg1 maturation 1960:1969 Fc N-glycan maturation 1948:1969 Fc N-glycan maturation 1948:1969 Thus, conformational restriction increases as Fc N-glycan maturation proceeds. 25551295 7 95 theme Modifying 1170:1178 arg1 Fc 1180:1181 Modifying Fc 1170:1181 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc 1170:1275 Modifying Fc with recombinantly expressed glycosyltransferases (Gnt1 and Gnt2) and UDP-[(13)C,(15)N]GlcNAc resulted in complete glycoform conversion as judged by mass spectrometry. 25551295 5 96 gly glycoforms 867:876 arg1 fragment 903:910 the immunoglobulin G1 fragment 881:910 the immunoglobulin G1 fragment crystallizable (Fc) 881:930 To address this technological gap, immature glycoforms of the immunoglobulin G1 fragment crystallizable (Fc) were isolated in a homogeneous state and enzymatically remodeled with [(13)C,(15)N]-N-acetylglucosamine (GlcNAc). 25551295 1 97 theme eukaryotic 217:226 arg1 modification 236:247 a common eukaryotic protein modification 208:247 a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues 208:379 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 25551295 1 97 theme eukaryotic 217:226 arg1 glycosylation 191:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation 169:203 Asparagine-linked (N) glycosylation is a common eukaryotic protein modification that affects protein folding, function, and stability through intramolecular interactions between N-glycan and polypeptide residues. 24840237 6 0 dep robustness 1359:1368 arg1 The 1355:1357 The 1355:1357 The 1355:1357 The robustness and selectivity of HILIC for N-glycan analyses allowed for method qualification. 24840237 4 1 theme N-glycans 956:964 arg1 separation 926:935 the baseline separation 913:935 the baseline separation of all typical mAb N-glycans 913:964 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 7 2 theme drug 1555:1558 arg1 release 1570:1576 clinical drug substance release 1546:1576 clinical drug substance release 1546:1576 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 1 3 theme chromatography 197:210 arg1 method 220:225 A highly robust hydrophilic interaction liquid chromatography (HILIC) method 150:225 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection 150:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 4 4 theme typical 944:950 arg1 N-glycans 956:964 all typical mAb N-glycans 940:964 all typical mAb N-glycans 940:964 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 0 5 theme related 127:133 arg1 glycoproteins 135:147 related glycoproteins 127:147 related glycoproteins 127:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 3 6 theme HILIC 637:641 arg1 column 643:648 a high-resolution, low-shedding HILIC column 605:648 a high-resolution, low-shedding HILIC column with acetonitrile 605:666 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 7 7 theme development 1489:1499 arg1 activities 1501:1510 bioprocess development activities 1478:1510 bioprocess development activities 1478:1510 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 5 8 dep such 1074:1077 arg1 maximizing 1307:1316 maximizing 1307:1316 maximizing sensitivity for trace-level species 1307:1352 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 2 9 theme mass 486:489 arg1 spectrometer 491:502 a quadrupole time-of-flight mass spectrometer 458:502 a quadrupole time-of-flight mass spectrometer 458:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 0 10 gly glycoproteins 135:147 arg1 glycoproteins 135:147 related glycoproteins 127:147 related glycoproteins 127:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 5 11 theme reproducible 1131:1142 arg1 profiles 1160:1167 reproducible chromatographic profiles 1131:1167 reproducible chromatographic profiles 1131:1167 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 12 theme conventional 1016:1027 arg1 formate 1038:1044 conventional ammonium formate 1016:1044 conventional ammonium formate 1016:1044 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 8 13 theme method 1608:1613 arg1 Application 1579:1589 Application 1579:1589 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), 1579:1689 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 0 14 theme method 59:64 arg1 application 16:26 application 16:26 application 16:26 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 0 14 theme method 59:64 arg1 Development 0:10 Development 0:10 Development 0:10 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 1 15 theme robust 159:164 arg1 method 220:225 A highly robust hydrophilic interaction liquid chromatography (HILIC) method 150:225 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection 150:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 2 16 theme separated 550:558 arg1 N-glycans 574:582 the separated, 2-AB-labeled N-glycans 546:582 the separated, 2-AB-labeled N-glycans 546:582 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 0 17 theme heightened 70:79 arg1 characterization 81:96 heightened characterization 70:96 heightened characterization of monoclonal antibodies and related glycoproteins 70:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 1 18 theme interaction 178:188 arg1 HILIC 213:217 HILIC 213:217 HILIC 213:217 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 1 18 theme interaction 178:188 arg1 chromatography 197:210 hydrophilic interaction liquid chromatography 166:210 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection 150:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 4 19 theme compatibility 853:865 arg1 combination 794:804 the combination 790:804 the combination of robust chromatographic performance and full compatibility and sensitivity with online MS 790:896 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 5 20 theme total 1183:1187 arg1 chromatograms 1201:1213 matching total ion current chromatograms 1174:1213 matching total ion current chromatograms 1174:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 7 21 theme substance 1560:1568 arg1 release 1570:1576 clinical drug substance release 1546:1576 clinical drug substance release 1546:1576 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 0 22 theme antibodies 112:121 arg1 characterization 81:96 heightened characterization 70:96 heightened characterization of monoclonal antibodies and related glycoproteins 70:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 5 23 theme current 1193:1199 arg1 chromatograms 1201:1213 matching total ion current chromatograms 1174:1213 matching total ion current chromatograms 1174:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 8 24 theme therapeutic 1662:1672 arg1 Rituxan 1679:1685 Rituxan 1679:1685 Rituxan(®) 1679:1688 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 8 24 theme therapeutic 1662:1672 arg1 mAb 1674:1676 a marketed therapeutic mAb 1651:1676 a marketed therapeutic mAb 1651:1676 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 4 25 theme chromatographic 816:830 arg1 performance 832:842 robust chromatographic performance 809:842 robust chromatographic performance 809:842 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 5 26 theme analyte 1253:1259 arg1 adduction 1261:1269 analyte adduction 1253:1269 analyte adduction 1253:1269 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 27 gly sialylated 1109:1118 arg1 N-glycans 1120:1128 sialylated N-glycans 1109:1128 sialylated N-glycans 1109:1128 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 2 28 theme quadrupole 460:469 arg1 spectrometer 491:502 a quadrupole time-of-flight mass spectrometer 458:502 a quadrupole time-of-flight mass spectrometer 458:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 4 29 dep separation 926:935 arg1 addition 901:908 addition 901:908 addition 901:908 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 5 30 theme minimal 1227:1233 arg1 signal 1235:1240 minimal signal 1227:1240 minimal signal splitting 1227:1250 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 2 31 theme mass 522:525 arg1 identifications 527:541 accurate mass identifications 513:541 accurate mass identifications of the separated, 2-AB-labeled N-glycans 513:582 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 4 32 theme baseline 917:924 arg1 separation 926:935 the baseline separation 913:935 the baseline separation of all typical mAb N-glycans 913:964 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 0 33 theme N-glycan 40:47 arg1 method 59:64 a robust N-glycan profiling method 31:64 a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins 31:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 5 34 theme sialylated 1109:1118 arg1 N-glycans 1120:1128 sialylated N-glycans 1109:1128 sialylated N-glycans 1109:1128 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 4 35 theme online 888:893 arg1 MS 895:896 online MS 888:896 online MS 888:896 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 2 36 theme Online 417:422 arg1 MS 449:450 MS 449:450 MS 449:450 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 2 36 theme Online 417:422 arg1 spectrometry 435:446 Online HILIC/mass spectrometry 417:446 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer 417:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 2 37 with spectrometry 435:446 arg1 spectrometer 491:502 a quadrupole time-of-flight mass spectrometer 458:502 a quadrupole time-of-flight mass spectrometer 458:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 5 38 theme optimal 1083:1089 arg1 order 1099:1103 optimal elution order 1083:1103 optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms 1083:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 4 39 with combination 794:804 arg1 MS 895:896 online MS 888:896 online MS 888:896 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 6 40 theme HILIC 1389:1393 arg1 selectivity 1374:1384 selectivity 1374:1384 selectivity 1374:1384 The robustness and selectivity of HILIC for N-glycan analyses allowed for method qualification. 24840237 6 40 theme HILIC 1389:1393 arg1 robustness 1359:1368 robustness 1359:1368 robustness 1359:1368 The robustness and selectivity of HILIC for N-glycan analyses allowed for method qualification. 24840237 1 41 theme spectrometric 268:280 arg1 detection 282:290 mass spectrometric detection 263:290 mass spectrometric detection 263:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 4 42 theme mAb 952:954 arg1 N-glycans 956:964 all typical mAb N-glycans 940:964 all typical mAb N-glycans 940:964 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 3 43 theme high-resolution 607:621 arg1 column 643:648 a high-resolution, low-shedding HILIC column 605:648 a high-resolution, low-shedding HILIC column with acetonitrile 605:666 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 5 44 theme TFA 978:980 arg1 use 971:973 The use 967:973 The use of TFA 967:980 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 1 45 theme liquid 190:195 arg1 HILIC 213:217 HILIC 213:217 HILIC 213:217 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 1 45 theme liquid 190:195 arg1 chromatography 197:210 hydrophilic interaction liquid chromatography 166:210 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection 150:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 7 46 theme clinical 1546:1553 arg1 release 1570:1576 clinical drug substance release 1546:1576 clinical drug substance release 1546:1576 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 0 47 theme glycoproteins 135:147 arg1 characterization 81:96 heightened characterization 70:96 heightened characterization of monoclonal antibodies and related glycoproteins 70:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 7 48 theme heightened 1513:1522 arg1 characterization 1524:1539 heightened characterization 1513:1539 heightened characterization 1513:1539 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 6 49 theme method 1429:1434 arg1 qualification 1436:1448 method qualification 1429:1448 method qualification 1429:1448 The robustness and selectivity of HILIC for N-glycan analyses allowed for method qualification. 24840237 5 50 theme matching 1174:1181 arg1 chromatograms 1201:1213 matching total ion current chromatograms 1174:1213 matching total ion current chromatograms 1174:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 51 theme distinct 991:998 arg1 advantages 1000:1009 distinct advantages 991:1009 distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species 991:1352 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 8 52 theme HILIC/MS 1599:1606 arg1 method 1608:1613 this HILIC/MS method 1594:1613 this HILIC/MS method 1594:1613 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 7 53 theme bioprocess 1478:1487 arg1 activities 1501:1510 bioprocess development activities 1478:1510 bioprocess development activities 1478:1510 The method is suitable for bioprocess development activities, heightened characterization, and clinical drug substance release. 24840237 6 54 theme N-glycan 1399:1406 arg1 analyses 1408:1415 N-glycan analyses 1399:1415 N-glycan analyses 1399:1415 The robustness and selectivity of HILIC for N-glycan analyses allowed for method qualification. 24840237 3 55 theme trifluoroacetic 709:723 arg1 TFA 731:733 TFA 731:733 TFA 731:733 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 55 theme trifluoroacetic 709:723 arg1 acid 725:728 trifluoroacetic acid 709:728 trifluoroacetic acid (TFA) 709:734 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 56 dep high-resolution 607:621 arg1 low-shedding 624:635 low-shedding 624:635 low-shedding 624:635 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 5 57 theme chromatographic 1144:1158 arg1 profiles 1160:1167 reproducible chromatographic profiles 1131:1167 reproducible chromatographic profiles 1131:1167 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 8 58 theme detailed 1622:1629 arg1 characterization 1631:1646 the detailed characterization 1618:1646 the detailed characterization of a marketed therapeutic mAb, Rituxan(®), 1618:1689 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 4 59 theme column 756:761 arg1 system 775:780 This column and solvent system 751:780 system 775:780 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 1 60 theme hydrophilic 166:176 arg1 HILIC 213:217 HILIC 213:217 HILIC 213:217 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 1 60 theme hydrophilic 166:176 arg1 chromatography 197:210 hydrophilic interaction liquid chromatography 166:210 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection 150:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 1 61 theme mAb 402:404 arg1 N-glycans 406:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 4 62 theme solvent 767:773 arg1 system 775:780 This column and solvent system 751:780 system 775:780 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 0 63 theme monoclonal 101:110 arg1 antibodies 112:121 monoclonal antibodies 101:121 monoclonal antibodies 101:121 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 1 64 theme -derivatized 389:400 arg1 N-glycans 406:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 5 65 theme mobile 1051:1056 arg1 phase 1058:1062 a mobile phase additive 1049:1071 a mobile phase additive 1049:1071 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 65 theme mobile 1051:1056 arg1 such 1074:1077 such 1074:1077 such 1074:1077 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 8 66 theme marketed 1653:1660 arg1 Rituxan 1679:1685 Rituxan 1679:1685 Rituxan(®) 1679:1688 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 8 66 theme marketed 1653:1660 arg1 mAb 1674:1676 a marketed therapeutic mAb 1651:1676 a marketed therapeutic mAb 1651:1676 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 4 67 theme full 848:851 arg1 compatibility 853:865 full compatibility 848:865 full compatibility 848:865 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 5 68 theme ion 1189:1191 arg1 chromatograms 1201:1213 matching total ion current chromatograms 1174:1213 matching total ion current chromatograms 1174:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 69 theme additive 1064:1071 arg1 phase 1058:1062 a mobile phase additive 1049:1071 a mobile phase additive 1049:1071 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 5 69 theme additive 1064:1071 arg1 such 1074:1077 such 1074:1077 such 1074:1077 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 2 70 theme N-glycans 574:582 arg1 identifications 527:541 accurate mass identifications 513:541 accurate mass identifications of the separated, 2-AB-labeled N-glycans 513:582 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 8 71 theme mAb 1674:1676 arg1 characterization 1631:1646 the detailed characterization 1618:1646 the detailed characterization of a marketed therapeutic mAb, Rituxan(®), 1618:1689 Application of this HILIC/MS method to the detailed characterization of a marketed therapeutic mAb, Rituxan(®), is described. 24840237 5 72 theme ammonium 1029:1036 arg1 formate 1038:1044 conventional ammonium formate 1016:1044 conventional ammonium formate 1016:1044 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 4 73 theme robust 809:814 arg1 performance 832:842 robust chromatographic performance 809:842 robust chromatographic performance 809:842 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 2 74 theme HILIC/mass 424:433 arg1 MS 449:450 MS 449:450 MS 449:450 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 2 74 theme HILIC/mass 424:433 arg1 spectrometry 435:446 Online HILIC/mass spectrometry 417:446 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer 417:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 3 75 with column 643:648 arg1 acetonitrile 655:666 acetonitrile 655:666 acetonitrile 655:666 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 2 76 theme time-of-flight 471:484 arg1 spectrometer 491:502 a quadrupole time-of-flight mass spectrometer 458:502 a quadrupole time-of-flight mass spectrometer 458:502 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 4 77 theme performance 832:842 arg1 combination 794:804 the combination 790:804 the combination of robust chromatographic performance and full compatibility and sensitivity with online MS 790:896 This column and solvent system ensures the combination of robust chromatographic performance and full compatibility and sensitivity with online MS in addition to the baseline separation of all typical mAb N-glycans. 24840237 0 78 theme robust 33:38 arg1 method 59:64 a robust N-glycan profiling method 31:64 a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins 31:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 5 79 theme elution 1091:1097 arg1 order 1099:1103 optimal elution order 1083:1103 optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms 1083:1213 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24840237 0 80 theme profiling 49:57 arg1 method 59:64 a robust N-glycan profiling method 31:64 a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins 31:147 Development and application of a robust N-glycan profiling method for heightened characterization of monoclonal antibodies and related glycoproteins. 24840237 1 81 theme released 353:360 arg1 N-glycans 406:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans 339:414 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 2 82 dep separated 550:558 arg1 2-AB-labeled 561:572 2-AB-labeled 561:572 2-AB-labeled 561:572 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 2 83 theme accurate 513:520 arg1 identifications 527:541 accurate mass identifications 513:541 accurate mass identifications of the separated, 2-AB-labeled N-glycans 513:582 Online HILIC/mass spectrometry (MS) with a quadrupole time-of-flight mass spectrometer provides accurate mass identifications of the separated, 2-AB-labeled N-glycans. 24840237 3 84 theme mobile 684:689 arg1 modifier 741:748 a modifier 739:748 a modifier 739:748 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 84 theme mobile 684:689 arg1 phases 691:696 mobile phases 684:696 mobile phases containing trifluoroacetic acid (TFA) 684:734 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 85 contain containing 698:707 arg2 acid 725:728 trifluoroacetic acid 709:728 trifluoroacetic acid (TFA) 709:734 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 85 contain containing 698:707 arg1 modifier 741:748 a modifier 739:748 a modifier 739:748 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 85 contain containing 698:707 arg2 TFA 731:733 TFA 731:733 TFA 731:733 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 3 85 contain containing 698:707 arg1 phases 691:696 mobile phases 684:696 mobile phases containing trifluoroacetic acid (TFA) 684:734 The method features a high-resolution, low-shedding HILIC column with acetonitrile and water-based mobile phases containing trifluoroacetic acid (TFA) as a modifier. 24840237 1 86 theme mass 263:266 arg1 detection 282:290 mass spectrometric detection 263:290 mass spectrometric detection 263:290 A highly robust hydrophilic interaction liquid chromatography (HILIC) method that involves both fluorescence and mass spectrometric detection was developed for profiling and characterizing enzymatically released and 2-aminobenzamide (2-AB)-derivatized mAb N-glycans. 24840237 5 87 theme trace-level 1334:1344 arg1 species 1346:1352 trace-level species 1334:1352 trace-level species 1334:1352 The use of TFA provided distinct advantages over conventional ammonium formate as a mobile phase additive, such as, optimal elution order for sialylated N-glycans, reproducible chromatographic profiles, and matching total ion current chromatograms, as well as minimal signal splitting, analyte adduction, and fragmentation during HILIC/MS, maximizing sensitivity for trace-level species. 24821760 9 0 theme specific 1593:1600 arg1 events 1616:1621 specific glycosylation events 1593:1621 specific glycosylation events 1593:1621 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 7 1 contain have 1212:1215 arg1 Ser3 1198:1201 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 7 1 contain have 1212:1215 arg2 effect 1246:1251 the largest themostabilizing effect 1217:1251 the largest themostabilizing effect 1217:1251 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 7 2 theme mannose 1174:1180 arg1 Ser3 1198:1201 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 3 3 from reesei 515:520 arg1 CBM 490:492 the Family 1 CBM 477:492 the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 477:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 9 4 theme glycosylation 1602:1614 arg1 events 1616:1621 specific glycosylation events 1593:1621 specific glycosylation events 1593:1621 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 6 5 from number 1026:1031 arg1 CBM 1057:1059 the CBM 1053:1059 the CBM 1053:1059 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 6 5 from number 1026:1031 arg1 site 1080:1083 the attachment site 1065:1083 the attachment site 1065:1083 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 1 6 theme lignocellulosic 170:184 arg1 biomass 186:192 lignocellulosic biomass 170:192 lignocellulosic biomass 170:192 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 8 7 from site 1311:1314 arg1 glycans 1295:1301 small glycans 1289:1301 small glycans at each site 1289:1314 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 1 8 theme carbohydrate-binding 255:274 arg1 CBMs 285:288 CBMs 285:288 CBMs 285:288 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 1 8 theme carbohydrate-binding 255:274 arg1 modules 276:282 Family 1 carbohydrate-binding modules 246:282 Family 1 carbohydrate-binding modules (CBMs) 246:289 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 5 9 theme CBMs 698:701 arg1 library 684:690 a library 682:690 a library of 20 CBMs 682:701 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 9 10 theme functional 1560:1569 arg1 studies 1571:1577 functional studies 1560:1577 functional studies 1560:1577 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 3 11 theme Family 522:527 arg1 cellobiohydrolase 531:547 Family 7 cellobiohydrolase 522:547 the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 499:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 9 12 theme glycoprotein 1533:1544 arg1 synthesis 1546:1554 chemical glycoprotein synthesis 1524:1554 chemical glycoprotein synthesis 1524:1554 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 0 13 theme Family 92:97 arg1 modules 122:128 Family 1 carbohydrate-binding modules 92:128 Family 1 carbohydrate-binding modules 92:128 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 8 14 theme affinity 1468:1475 arg1 enhancement 1477:1487 the highest affinity enhancement 1456:1487 the highest affinity enhancement of 7.4-fold 1456:1499 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 7 15 theme disaccharide 1182:1193 arg1 Ser3 1198:1201 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 8 16 with glycoform 1387:1395 arg1 mannose 1411:1417 a single mannose 1402:1417 a single mannose at each of three positions 1402:1444 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 0 17 theme modules 122:128 arg1 cellulose 62:70 cellulose 62:70 cellulose 62:70 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 0 17 theme modules 122:128 arg1 stability 48:56 stability 48:56 stability 48:56 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 7 18 theme at 1195:1196 arg1 Ser3 1198:1201 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 2 19 gly glycosylation 378:390 arg1 CBM 395:397 CBM function 395:406 CBM function 395:406 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 1 20 from turnover 158:165 arg1 nature 197:202 nature 197:202 nature 197:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 3 21 theme Trichoderma 503:513 arg1 reesei 515:520 the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 499:576 the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 499:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 7 22 theme largest 1221:1227 arg1 effect 1246:1251 the largest themostabilizing effect 1217:1251 the largest themostabilizing effect 1217:1251 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 1 23 theme biomass 186:192 arg1 turnover 158:165 biological turnover 147:165 biological turnover of lignocellulosic biomass in nature 147:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 0 24 theme carbohydrate-binding 101:120 arg1 modules 122:128 Family 1 carbohydrate-binding modules 92:128 Family 1 carbohydrate-binding modules 92:128 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 9 25 theme beneficial 1639:1648 arg1 properties 1650:1659 multiple beneficial properties 1630:1659 multiple beneficial properties 1630:1659 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 1 26 from majority 135:142 arg1 nature 197:202 nature 197:202 nature 197:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 7 27 gly glycoforms 1144:1153 arg1 CBM 1140:1142 CBM glycoforms 1140:1153 CBM glycoforms 1140:1153 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 2 28 theme glycosylation 378:390 arg1 effects 367:373 the effects 363:373 the effects of glycosylation on CBM function 363:406 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 6 29 theme major 913:917 arg1 changes 934:940 major conformational changes 913:940 major conformational changes 913:940 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 6 30 theme mannose 1036:1042 arg1 units 1044:1048 mannose units 1036:1048 mannose units 1036:1048 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 8 31 theme highest 1460:1466 arg1 enhancement 1477:1487 the highest affinity enhancement 1456:1487 the highest affinity enhancement of 7.4-fold 1456:1499 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 7 32 theme CBM 1140:1142 arg1 glycoforms 1144:1153 CBM glycoforms 1140:1153 CBM glycoforms 1140:1153 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 4 33 gly glycosylated 626:637 arg1 CBMs 648:651 glycosylated Family 1 CBMs 626:651 glycosylated Family 1 CBMs 626:651 To enable this work, a procedure to synthesize glycosylated Family 1 CBMs was developed. 24821760 3 34 theme glycosylation 558:570 arg1 sites 572:576 three glycosylation sites 552:576 three glycosylation sites 552:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 3 35 from sites 572:576 arg1 cellobiohydrolase 531:547 Family 7 cellobiohydrolase 522:547 the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 499:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 5 36 theme thermostability 830:844 arg1 comparison 771:780 comparison 771:780 comparison of binding affinity, proteolytic stability, and thermostability 771:844 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 8 37 theme 7.4-fold 1492:1499 arg1 enhancement 1477:1487 the highest affinity enhancement 1456:1487 the highest affinity enhancement of 7.4-fold 1456:1499 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 7 38 dep °C 1164:1165 arg1 up 1155:1156 up 1155:1156 up 1155:1156 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 7 39 from Ser3 1198:1201 arg1 at 1195:1196 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 7 39 from Ser3 1198:1201 arg1 mannose 1174:1180 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 7 39 from Ser3 1198:1201 arg1 disaccharide 1182:1193 a mannose disaccharide at Ser3 1172:1201 a mannose disaccharide at Ser3 1172:1201 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 8 40 theme higher 1326:1331 arg1 affinities 1341:1350 higher binding affinities 1326:1350 higher binding affinities for crystalline cellulose 1326:1376 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 3 41 theme O-mannosylation 445:459 arg1 effects 434:440 the effects 430:440 the effects of O-mannosylation 430:459 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 0 42 theme O-glycosylation 15:29 arg1 Specificity 0:10 Specificity 0:10 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 0:129 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 6 43 theme conformational 919:932 arg1 changes 934:940 major conformational changes 913:940 major conformational changes 913:940 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 6 44 theme units 1044:1048 arg1 number 1026:1031 the number 1022:1031 the number of mannose units on the CBM and the attachment site 1022:1083 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 5 45 theme affinity 793:800 arg1 comparison 771:780 comparison 771:780 comparison of binding affinity, proteolytic stability, and thermostability 771:844 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 7 46 theme themostabilizing 1229:1244 arg1 effect 1246:1251 the largest themostabilizing effect 1217:1251 the largest themostabilizing effect 1217:1251 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 3 47 dep reesei 515:520 arg1 cellobiohydrolase 531:547 Family 7 cellobiohydrolase 522:547 the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 499:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 2 48 gly glycosylated 345:356 arg1 CBMs 336:339 Family 1 CBMs 327:339 Family 1 CBMs 327:339 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 6 49 theme attachment 1069:1078 arg1 site 1080:1083 the attachment site 1065:1083 the attachment site 1065:1083 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 4 50 theme glycosylated 626:637 arg1 CBMs 648:651 glycosylated Family 1 CBMs 626:651 glycosylated Family 1 CBMs 626:651 To enable this work, a procedure to synthesize glycosylated Family 1 CBMs was developed. 24821760 5 51 theme stability 815:823 arg1 comparison 771:780 comparison 771:780 comparison of binding affinity, proteolytic stability, and thermostability 771:844 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 8 52 theme single 1404:1409 arg1 mannose 1411:1417 a single mannose 1402:1417 a single mannose at each of three positions 1402:1444 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 0 53 dep stability 48:56 arg1 affinity 80:87 binding affinity 72:87 binding affinity 72:87 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 0 53 dep stability 48:56 arg1 the 44:46 the 44:46 the 44:46 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 2 54 theme CBM 395:397 arg1 function 399:406 CBM function 395:406 CBM function 395:406 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 8 55 theme small 1289:1293 arg1 glycans 1295:1301 small glycans 1289:1301 small glycans at each site 1289:1314 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 8 56 theme crystalline 1356:1366 arg1 cellulose 1368:1376 crystalline cellulose 1356:1376 crystalline cellulose 1356:1376 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 2 57 from effects 367:373 arg1 function 399:406 CBM function 395:406 CBM function 395:406 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 3 58 gly glycosylation 558:570 arg2 sites 572:576 three glycosylation sites 552:576 three glycosylation sites 552:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 3 58 gly glycosylation 558:570 arg2 three 552:556 three 552:556 three 552:556 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 9 59 gly glycoprotein 1533:1544 arg1 glycoprotein 1533:1544 chemical glycoprotein synthesis 1524:1554 chemical glycoprotein synthesis 1524:1554 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 9 60 theme Family 1664:1669 arg1 CBMs 1673:1676 Family 1 CBMs 1664:1676 Family 1 CBMs 1664:1676 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 8 61 with glycoforms 1273:1282 arg1 glycans 1295:1301 small glycans 1289:1301 small glycans at each site 1289:1314 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 4 62 dep procedure 602:610 arg1 synthesize 615:624 synthesize 615:624 to synthesize glycosylated Family 1 CBMs 612:651 To enable this work, a procedure to synthesize glycosylated Family 1 CBMs was developed. 24821760 3 63 theme Family 481:486 arg1 CBM 490:492 the Family 1 CBM 477:492 the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites 477:576 Here, the effects of O-mannosylation are examined on the Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase at three glycosylation sites. 24821760 8 64 theme binding 1333:1339 arg1 affinities 1341:1350 higher binding affinities 1326:1350 higher binding affinities for crystalline cellulose 1326:1376 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 9 65 theme chemical 1524:1531 arg1 synthesis 1546:1554 chemical glycoprotein synthesis 1524:1554 chemical glycoprotein synthesis 1524:1554 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 9 66 theme multiple 1630:1637 arg1 properties 1650:1659 multiple beneficial properties 1630:1659 multiple beneficial properties 1630:1659 Overall, by combining chemical glycoprotein synthesis and functional studies, we show that specific glycosylation events confer multiple beneficial properties on Family 1 CBMs. 24821760 6 67 dep 50-fold 1001:1007 arg1 up 995:996 up 995:996 up 995:996 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 1 68 from nature 197:202 arg1 majority 135:142 The majority 131:142 The majority of biological turnover of lignocellulosic biomass in nature 131:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 2 69 theme Family 327:332 arg1 CBMs 336:339 Family 1 CBMs 327:339 Family 1 CBMs 327:339 Family 1 CBMs are glycosylated, but the effects of glycosylation on CBM function remain unknown. 24821760 8 70 from each 1422:1425 arg1 mannose 1411:1417 a single mannose 1402:1417 a single mannose at each of three positions 1402:1444 Interestingly, the glycoforms with small glycans at each site displayed higher binding affinities for crystalline cellulose, and the glycoform with a single mannose at each of three positions conferred the highest affinity enhancement of 7.4-fold. 24821760 0 71 theme binding 72:78 arg1 affinity 80:87 binding affinity 72:87 binding affinity 72:87 Specificity of O-glycosylation in enhancing the stability and cellulose binding affinity of Family 1 carbohydrate-binding modules. 24821760 5 72 theme proteolytic 803:813 arg1 stability 815:823 proteolytic stability 803:823 proteolytic stability 803:823 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 5 73 theme binding 785:791 arg1 affinity 793:800 binding affinity 785:800 binding affinity 785:800 Subsequently, a library of 20 CBMs was synthesized with mono-, di-, or trisaccharides at each site for comparison of binding affinity, proteolytic stability, and thermostability. 24821760 1 74 theme biological 147:156 arg1 turnover 158:165 biological turnover 147:165 biological turnover of lignocellulosic biomass in nature 147:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 4 75 theme Family 639:644 arg1 CBMs 648:651 glycosylated Family 1 CBMs 626:651 glycosylated Family 1 CBMs 626:651 To enable this work, a procedure to synthesize glycosylated Family 1 CBMs was developed. 24821760 6 76 theme cleavage 975:982 arg1 resistance 984:993 the thermolysin cleavage resistance 959:993 the thermolysin cleavage resistance up to 50-fold 959:1007 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 7 77 theme glycoforms 1144:1153 arg1 thermostability 1121:1135 the thermostability 1117:1135 the thermostability of CBM glycoforms 1117:1153 O-Mannosylation also increases the thermostability of CBM glycoforms up to 16 °C, and a mannose disaccharide at Ser3 seems to have the largest themostabilizing effect. 24821760 6 78 theme CBM 879:881 arg1 mannosylation 883:895 CBM mannosylation 879:895 CBM mannosylation 879:895 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 6 79 theme thermolysin 963:973 arg1 resistance 984:993 the thermolysin cleavage resistance 959:993 the thermolysin cleavage resistance up to 50-fold 959:1007 The results show that, although CBM mannosylation does not induce major conformational changes, it can increase the thermolysin cleavage resistance up to 50-fold depending on the number of mannose units on the CBM and the attachment site. 24821760 1 80 theme turnover 158:165 arg1 majority 135:142 The majority 131:142 The majority of biological turnover of lignocellulosic biomass in nature 131:202 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 1 81 theme Family 246:251 arg1 CBMs 285:288 CBMs 285:288 CBMs 285:288 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24821760 1 81 theme Family 246:251 arg1 modules 276:282 Family 1 carbohydrate-binding modules 246:282 Family 1 carbohydrate-binding modules (CBMs) 246:289 The majority of biological turnover of lignocellulosic biomass in nature is conducted by fungi, which commonly use Family 1 carbohydrate-binding modules (CBMs) for targeting enzymes to cellulose. 24663386 6 0 theme type 1360:1363 arg1 antigen 1367:1373 blood type A antigen 1354:1373 blood type A antigen 1354:1373 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 1 theme Lewis 1425:1429 arg1 antigen 1433:1439 sialyl Lewis X antigen 1418:1439 sialyl Lewis X antigen 1418:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 2 2 from controls 199:206 arg1 sera 176:179 human sera 170:179 human sera from four healthy controls 170:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 2 2 from controls 199:206 arg1 structures 143:152 The structures 139:152 The structures of O-glycans in human sera from four healthy controls 139:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 6 3 dep glycans 1224:1230 arg1 ii 1209:1210 ii 1209:1210 ii 1209:1210 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 4 theme X 1404:1404 arg1 antigen 1406:1412 Lewis X antigen 1398:1412 Lewis X antigen 1398:1412 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 0 5 link O-linked 31:38 arg1 oligosaccharides 40:55 O-linked oligosaccharides 31:55 O-linked oligosaccharides in human serum 31:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 7 6 from antigen 1598:1604 arg1 controls 1617:1624 healthy controls 1609:1624 healthy controls 1609:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 5 7 gly sialylated 885:894 arg1 glycans 896:902 other sialylated glycans 879:902 other sialylated glycans 879:902 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 6 8 theme Galβ1-4GlcNAcβ1-6 1174:1190 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 8 theme Galβ1-4GlcNAcβ1-6 1174:1190 arg1 GalNAc 1200:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 9 theme sialyl 1240:1245 arg1 antigen 1250:1256 sialyl Tn antigen 1240:1256 sialyl Tn antigen 1240:1256 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 10 from kinds 1512:1516 arg1 controls 1617:1624 healthy controls 1609:1624 healthy controls 1609:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 0 11 from serum 66:70 arg1 analysis 19:26 Precise structural analysis 0:26 Precise structural analysis of O-linked oligosaccharides in human serum 0:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 6 12 theme glycan 1166:1171 arg1 glycans 1316:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans 1150:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen 1150:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 13 dep markers 1546:1552 arg1 antigen 1571:1577 sialyl Tn antigen 1561:1577 clinically applied tumor markers namely sialyl Tn antigen 1521:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 4 14 theme neutral 692:698 arg1 glycans 700:706 neutral glycans 692:706 neutral glycans 692:706 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 5 15 theme NeuAcα2-3Galβ1-3 1004:1019 arg1 GalNAc 1021:1026 NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 977:1026 NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 977:1026 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 1 16 theme useful 117:122 arg1 markers 130:136 novel and useful tumor markers 107:136 novel and useful tumor markers 107:136 O-glycans are suitable targets as novel and useful tumor markers. 24663386 6 17 theme sialylated 1213:1222 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 17 theme sialylated 1213:1222 arg1 glycans 1224:1230 (ii) sialylated glycans 1208:1230 (ii) sialylated glycans 1208:1230 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 2 18 theme reference 246:254 arg1 database 265:272 the reference O-glycan database 242:272 the reference O-glycan database 242:272 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 5 19 theme sialylated 885:894 arg1 glycans 896:902 other sialylated glycans 879:902 other sialylated glycans 879:902 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 7 20 theme Lewis 1590:1594 arg1 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 1 21 theme tumor 124:128 arg1 markers 130:136 novel and useful tumor markers 107:136 novel and useful tumor markers 107:136 O-glycans are suitable targets as novel and useful tumor markers. 24663386 6 22 dep glycan 1166:1171 arg1 i 1151:1151 i 1151:1151 i 1151:1151 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 3 23 theme hydrazine 312:320 arg1 treatment 322:330 hydrazine treatment 312:330 hydrazine treatment followed by fluorescent labeling with aminopyridine 312:382 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 0 24 from analysis 19:26 arg1 serum 66:70 human serum 60:70 human serum 60:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 8 25 theme relative 1768:1775 arg1 glycans 1794:1800 these glycans 1788:1800 these glycans 1788:1800 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 25 theme relative 1768:1775 arg1 % 1813:1813 <0.01% 1808:1813 <0.01% 1808:1813 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 25 theme relative 1768:1775 arg1 amounts 1777:1783 the relative amounts 1764:1783 the relative amounts of these glycans 1764:1800 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 26 theme glycans 1794:1800 arg1 glycans 1794:1800 these glycans 1788:1800 these glycans 1788:1800 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 26 theme glycans 1794:1800 arg1 % 1813:1813 <0.01% 1808:1813 <0.01% 1808:1813 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 26 theme glycans 1794:1800 arg1 amounts 1777:1783 the relative amounts 1764:1783 the relative amounts of these glycans 1764:1800 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 5 27 theme major 856:860 arg1 components 862:871 major components 856:871 major components while other sialylated glycans 856:902 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 6 28 dep glycans 1459:1465 arg1 iv 1446:1447 iv 1446:1447 iv 1446:1447 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 9 29 theme detailed 1822:1829 arg1 profiles 1840:1847 These detailed O-glycan profiles 1816:1847 These detailed O-glycan profiles 1816:1847 These detailed O-glycan profiles will help to find novel carbohydrate tumor markers. 24663386 6 30 theme trisialylated 1272:1284 arg1 structures 1286:1295 trisialylated structures 1272:1295 trisialylated structures 1272:1295 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 3 31 theme fluorescent 344:354 arg1 labeling 356:363 fluorescent labeling 344:363 fluorescent labeling with aminopyridine 344:382 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 5 32 theme ∼1-2 1077:1080 arg1 % 1081:1081 ∼1-2% 1077:1081 ∼1-2% 1077:1081 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 7 33 located present 1651:1657 arg1 % 1670:1670 ∼0.1-0.2% 1662:1670 ∼0.1-0.2% of the total 1662:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 33 located present 1651:1657 arg2 kinds 1512:1516 Two kinds 1508:1516 Two kinds of clinically applied tumor markers namely sialyl Tn antigen 1508:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 33 located present 1651:1657 arg2 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 33 located present 1651:1657 arg1 total 1679:1683 total 1679:1683 total 1679:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 34 theme tumor 1540:1544 arg1 markers 1546:1552 clinically applied tumor markers 1521:1552 clinically applied tumor markers namely sialyl Tn antigen 1521:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 0 35 theme Precise 0:6 arg1 analysis 19:26 Precise structural analysis 0:26 Precise structural analysis of O-linked oligosaccharides in human serum 0:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 9 36 theme carbohydrate 1873:1884 arg1 markers 1892:1898 novel carbohydrate tumor markers 1867:1898 novel carbohydrate tumor markers 1867:1898 These detailed O-glycan profiles will help to find novel carbohydrate tumor markers. 24663386 6 37 theme type 1338:1341 arg1 antigen 1345:1351 blood type H antigen 1332:1351 blood type H antigen 1332:1351 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 4 38 attach present 588:594 arg1 % 613:613 more than 0.01% 599:613 more than 0.01% of the total O-glycans 599:636 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 4 38 attach present 588:594 arg2 O-glycans 577:585 O-glycans 577:585 O-glycans 577:585 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 4 38 attach present 588:594 arg1 O-glycans 628:636 the total O-glycans 618:636 the total O-glycans 618:636 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 1 39 theme novel 107:111 arg1 markers 130:136 novel and useful tumor markers 107:136 novel and useful tumor markers 107:136 O-glycans are suitable targets as novel and useful tumor markers. 24663386 3 40 theme enzymatic 430:438 arg1 digestion 440:448 enzymatic digestion 430:448 enzymatic digestion 430:448 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 4 41 theme present 588:594 arg1 O-glycans 577:585 O-glycans 577:585 O-glycans 577:585 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 2 42 theme human 170:174 arg1 sera 176:179 human sera 170:179 human sera from four healthy controls 170:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 0 43 theme O-linked 31:38 arg1 oligosaccharides 40:55 O-linked oligosaccharides 31:55 O-linked oligosaccharides in human serum 31:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 2 44 theme O-glycans 157:165 arg1 structures 143:152 The structures 139:152 The structures of O-glycans in human sera from four healthy controls 139:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 6 45 theme fucosylated 1304:1314 arg1 glycans 1316:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans 1150:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen 1150:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 46 theme minor 1089:1093 arg1 glycans 1095:1101 Very minor glycans 1084:1101 Very minor glycans accounting for ∼0.01-0.1% of the total 1084:1140 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 2 47 theme healthy 191:197 arg1 controls 199:206 four healthy controls 186:206 four healthy controls 186:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 6 48 theme type 1382:1385 arg1 antigen 1389:1395 blood type B antigen 1376:1395 blood type B antigen 1376:1395 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 3 49 theme synthesized 525:535 arg1 oligosaccharides 546:561 newly synthesized sulfated oligosaccharides 519:561 newly synthesized sulfated oligosaccharides as standards 519:574 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 4 50 theme total 622:626 arg1 O-glycans 628:636 the total O-glycans 618:636 the total O-glycans 618:636 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 8 51 theme other 1695:1699 arg1 markers 1701:1707 other markers 1695:1707 other markers such as CA19-9 and DU-PAN-2 1695:1735 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 51 theme other 1695:1699 arg1 CA19-9 1717:1722 CA19-9 1717:1722 CA19-9 1717:1722 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 8 51 theme other 1695:1699 arg1 DU-PAN-2 1728:1735 DU-PAN-2 1728:1735 DU-PAN-2 1728:1735 However, other markers such as CA19-9 and DU-PAN-2 were not found, suggesting the relative amounts of these glycans to be <0.01%. 24663386 6 52 theme A 1365:1365 arg1 antigen 1367:1373 blood type A antigen 1354:1373 blood type A antigen 1354:1373 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 53 theme blood 1354:1358 arg1 antigen 1367:1373 blood type A antigen 1354:1373 blood type A antigen 1354:1373 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 54 theme total 1136:1140 arg1 % 1127:1127 ∼0.01-0.1% 1118:1127 ∼0.01-0.1% of the total 1118:1140 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 54 theme total 1136:1140 arg1 total 1136:1140 total 1136:1140 total 1136:1140 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 3 55 theme oligosaccharides 546:561 arg1 spectrometry 459:470 mass spectrometry 454:470 mass spectrometry (MS) 454:475 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 55 theme oligosaccharides 546:561 arg1 use 512:514 the use 508:514 the use of newly synthesized sulfated oligosaccharides as standards 508:574 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 55 theme oligosaccharides 546:561 arg1 MS 473:474 MS 473:474 MS 473:474 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 55 theme oligosaccharides 546:561 arg1 mapping 421:427 two-dimensional mapping 405:427 two-dimensional mapping 405:427 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 55 theme oligosaccharides 546:561 arg1 digestion 440:448 enzymatic digestion 430:448 enzymatic digestion 430:448 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 6 56 theme sialyl 1418:1423 arg1 antigen 1433:1439 sialyl Lewis X antigen 1418:1439 sialyl Lewis X antigen 1418:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 57 theme Galβ1-3 1192:1198 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 57 theme Galβ1-3 1192:1198 arg1 GalNAc 1200:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 58 gly sialylated 1213:1222 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 58 gly sialylated 1213:1222 arg1 glycans 1224:1230 (ii) sialylated glycans 1208:1230 (ii) sialylated glycans 1208:1230 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 59 theme healthy 1609:1615 arg1 controls 1617:1624 healthy controls 1609:1624 healthy controls 1609:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 6 60 theme Lewis 1398:1402 arg1 antigen 1406:1412 Lewis X antigen 1398:1412 Lewis X antigen 1398:1412 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 61 attach present 1651:1657 arg1 % 1670:1670 ∼0.1-0.2% 1662:1670 ∼0.1-0.2% of the total 1662:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 61 attach present 1651:1657 arg2 kinds 1512:1516 Two kinds 1508:1516 Two kinds of clinically applied tumor markers namely sialyl Tn antigen 1508:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 61 attach present 1651:1657 arg2 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 61 attach present 1651:1657 arg1 total 1679:1683 total 1679:1683 total 1679:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 6 62 theme Tn 1247:1248 arg1 antigen 1250:1256 sialyl Tn antigen 1240:1256 sialyl Tn antigen 1240:1256 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 5 63 theme minor 1044:1048 arg1 components 1050:1059 relatively minor components 1033:1059 relatively minor components 1033:1059 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 3 64 with labeling 356:363 arg1 aminopyridine 370:382 aminopyridine 370:382 aminopyridine 370:382 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 6 65 theme sulfated 1450:1457 arg1 glycans 1459:1465 (iv) sulfated glycans 1445:1465 (iv) sulfated glycans 1445:1465 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 2 66 from structures 143:152 arg1 sera 176:179 human sera 170:179 human sera from four healthy controls 170:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 2 66 from structures 143:152 arg1 controls 199:206 four healthy controls 186:206 four healthy controls 186:206 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 6 67 theme neutral 1158:1164 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 67 theme neutral 1158:1164 arg1 GalNAc 1200:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 67 theme neutral 1158:1164 arg1 glycans 1224:1230 (ii) sialylated glycans 1208:1230 (ii) sialylated glycans 1208:1230 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 2 68 theme O-glycan 256:263 arg1 database 265:272 the reference O-glycan database 242:272 the reference O-glycan database 242:272 The structures of O-glycans in human sera from four healthy controls were precisely analyzed to obtain the reference O-glycan database. 24663386 0 69 theme oligosaccharides 40:55 arg1 analysis 19:26 Precise structural analysis 0:26 Precise structural analysis of O-linked oligosaccharides in human serum 0:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 7 70 dep kinds 1512:1516 arg1 sera 1626:1629 sera 1626:1629 sera 1626:1629 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 4 71 theme glycans 700:706 arg1 kinds 683:687 2 kinds 681:687 2 kinds of neutral glycans 681:706 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 4 71 theme glycans 700:706 arg1 kinds 661:665 18 kinds 658:665 18 kinds of acidic 658:675 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 7 72 theme X 1596:1596 arg1 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 4 73 theme acidic 670:675 arg1 kinds 683:687 2 kinds 681:687 2 kinds of neutral glycans 681:706 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 4 73 theme acidic 670:675 arg1 kinds 661:665 18 kinds 658:665 18 kinds of acidic 658:675 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 7 74 theme Tn 1568:1569 arg1 antigen 1571:1577 sialyl Tn antigen 1561:1577 clinically applied tumor markers namely sialyl Tn antigen 1521:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 6 75 theme X 1431:1431 arg1 antigen 1433:1439 sialyl Lewis X antigen 1418:1439 sialyl Lewis X antigen 1418:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 76 theme sialyl 1583:1588 arg1 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 4 77 from % 613:613 arg1 present 588:594 present 588:594 present 588:594 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 5 78 theme NeuAcα2-6 913:921 arg1 GalNAc 923:928 Galβ1-3(NeuAcα2-6)GalNAc 905:928 Galβ1-3(NeuAcα2-6)GalNAc 905:928 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 6 79 contain having 1233:1238 arg2 agalacto 1259:1266 agalacto 1259:1266 agalacto 1259:1266 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 79 contain having 1233:1238 arg1 GalNAc 1200:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc 1174:1205 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 79 contain having 1233:1238 arg2 structures 1286:1295 trisialylated structures 1272:1295 trisialylated structures 1272:1295 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 79 contain having 1233:1238 arg1 glycans 1224:1230 (ii) sialylated glycans 1208:1230 (ii) sialylated glycans 1208:1230 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 79 contain having 1233:1238 arg1 glycan 1166:1171 the neutral glycan 1154:1171 the neutral glycan 1154:1171 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 79 contain having 1233:1238 arg2 antigen 1250:1256 sialyl Tn antigen 1240:1256 sialyl Tn antigen 1240:1256 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 9 80 theme O-glycan 1831:1838 arg1 profiles 1840:1847 These detailed O-glycan profiles 1816:1847 These detailed O-glycan profiles 1816:1847 These detailed O-glycan profiles will help to find novel carbohydrate tumor markers. 24663386 6 81 theme 3'-sulfo 1487:1494 arg1 structures 1496:1505 6-sulfo and 3'-sulfo structures 1475:1505 6-sulfo and 3'-sulfo structures 1475:1505 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 6 82 gly fucosylated 1304:1314 arg1 glycans 1316:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans 1150:1322 (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen 1150:1439 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 5 83 theme other 879:883 arg1 glycans 896:902 other sialylated glycans 879:902 other sialylated glycans 879:902 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 0 84 from oligosaccharides 40:55 arg1 serum 66:70 human serum 60:70 human serum 60:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 0 85 theme structural 8:17 arg1 analysis 19:26 Precise structural analysis 0:26 Precise structural analysis of O-linked oligosaccharides in human serum 0:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 6 86 theme 6-sulfo 1475:1481 arg1 structures 1496:1505 6-sulfo and 3'-sulfo structures 1475:1505 6-sulfo and 3'-sulfo structures 1475:1505 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 1 87 theme suitable 87:94 arg1 O-glycans 73:81 O-glycans 73:81 O-glycans 73:81 O-glycans are suitable targets as novel and useful tumor markers. 24663386 1 87 theme suitable 87:94 arg1 targets 96:102 suitable targets 87:102 suitable targets as novel and useful tumor markers 87:136 O-glycans are suitable targets as novel and useful tumor markers. 24663386 7 88 theme markers 1546:1552 arg1 kinds 1512:1516 Two kinds 1508:1516 Two kinds of clinically applied tumor markers namely sialyl Tn antigen 1508:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 88 theme markers 1546:1552 arg1 antigen 1598:1604 sialyl Lewis X antigen 1583:1604 sialyl Lewis X antigen in healthy controls 1583:1624 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 89 from % 1670:1670 arg1 present 1651:1657 present 1651:1657 present 1651:1657 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 3 90 theme two-dimensional 405:419 arg1 mapping 421:427 two-dimensional mapping 405:427 two-dimensional mapping 405:427 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 91 theme mass 454:457 arg1 spectrometry 459:470 mass spectrometry 454:470 mass spectrometry (MS) 454:475 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 3 91 theme mass 454:457 arg1 MS 473:474 MS 473:474 MS 473:474 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 24663386 6 92 theme H 1343:1343 arg1 antigen 1345:1351 blood type H antigen 1332:1351 blood type H antigen 1332:1351 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 5 93 theme Galβ1-3 905:911 arg1 GalNAc 923:928 Galβ1-3(NeuAcα2-6)GalNAc 905:928 Galβ1-3(NeuAcα2-6)GalNAc 905:928 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 7 94 theme applied 1532:1538 arg1 markers 1546:1552 clinically applied tumor markers 1521:1552 clinically applied tumor markers namely sialyl Tn antigen 1521:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 9 95 theme novel 1867:1871 arg1 markers 1892:1898 novel carbohydrate tumor markers 1867:1898 novel carbohydrate tumor markers 1867:1898 These detailed O-glycan profiles will help to find novel carbohydrate tumor markers. 24663386 6 96 theme blood 1332:1336 arg1 antigen 1345:1351 blood type H antigen 1332:1351 blood type H antigen 1332:1351 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 7 97 theme sialyl 1561:1566 arg1 antigen 1571:1577 sialyl Tn antigen 1561:1577 clinically applied tumor markers namely sialyl Tn antigen 1521:1577 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 6 98 gly trisialylated 1272:1284 arg1 structures 1286:1295 trisialylated structures 1272:1295 trisialylated structures 1272:1295 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 9 99 theme tumor 1886:1890 arg1 markers 1892:1898 novel carbohydrate tumor markers 1867:1898 novel carbohydrate tumor markers 1867:1898 These detailed O-glycan profiles will help to find novel carbohydrate tumor markers. 24663386 0 100 theme human 60:64 arg1 serum 66:70 human serum 60:70 human serum 60:70 Precise structural analysis of O-linked oligosaccharides in human serum. 24663386 5 101 theme NeuAcα2-3Galβ1-4GlcNAcβ1-6 977:1002 arg1 GalNAc 1021:1026 NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 977:1026 NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 977:1026 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 6 102 theme B 1387:1387 arg1 antigen 1389:1395 blood type B antigen 1376:1395 blood type B antigen 1376:1395 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 5 103 theme Galβ1-4GlcNAcβ1-6 931:947 arg1 GalNAc 966:971 Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 931:971 Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 931:971 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 7 104 theme total 1679:1683 arg1 % 1670:1670 ∼0.1-0.2% 1662:1670 ∼0.1-0.2% of the total 1662:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 7 104 theme total 1679:1683 arg1 total 1679:1683 total 1679:1683 total 1679:1683 Two kinds of clinically applied tumor markers namely sialyl Tn antigen and sialyl Lewis X antigen in healthy controls sera were revealed to be present at ∼0.1-0.2% of the total. 24663386 6 105 theme blood 1376:1380 arg1 antigen 1389:1395 blood type B antigen 1376:1395 blood type B antigen 1376:1395 Very minor glycans accounting for ∼0.01-0.1% of the total include (i) the neutral glycan, Galβ1-4GlcNAcβ1-6(Galβ1-3)GalNAc, (ii) sialylated glycans, having sialyl Tn antigen, agalacto and trisialylated structures, (iii) fucosylated glycans forming blood type H antigen, blood type A antigen, blood type B antigen, Lewis X antigen and sialyl Lewis X antigen and (iv) sulfated glycans, having 6-sulfo and 3'-sulfo structures. 24663386 5 106 theme NeuAcα2-3Galβ1-3 949:964 arg1 GalNAc 966:971 Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 931:971 Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc 931:971 NeuAcα2-3Galβ1-3N-acetylgalactosamine (GalNAc) (61-64%), NeuAcα2-3Galβ1-3(NeuAcα2-6)GalNAc (15-26%) and Galβ1-3GalNAc (6-14%) were major components while other sialylated glycans, Galβ1-3(NeuAcα2-6)GalNAc, Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc and NeuAcα2-3Galβ1-4GlcNAcβ1-6(NeuAcα2-3Galβ1-3)GalNAc were relatively minor components, accounting for ∼1-2%. 24663386 4 107 theme O-glycans 628:636 arg1 % 613:613 more than 0.01% 599:613 more than 0.01% of the total O-glycans 599:636 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 4 107 theme O-glycans 628:636 arg1 O-glycans 628:636 the total O-glycans 618:636 the total O-glycans 618:636 O-glycans, present at more than 0.01% of the total O-glycans, were analyzed, and 18 kinds of acidic and 2 kinds of neutral glycans were identified. 24663386 3 108 theme sulfated 537:544 arg1 oligosaccharides 546:561 newly synthesized sulfated oligosaccharides 519:561 newly synthesized sulfated oligosaccharides as standards 519:574 O-glycans were prepared from sera by hydrazine treatment followed by fluorescent labeling with aminopyridine and identified using two-dimensional mapping, enzymatic digestion and mass spectrometry (MS) together with methanolysis and the use of newly synthesized sulfated oligosaccharides as standards. 28994411 2 0 theme glycan 483:488 arg1 forms 490:494 high-mannose glycan forms 470:494 high-mannose glycan forms only 470:499 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 3 1 theme processed 706:714 arg1 glycosylation 727:739 fully processed and native glycosylation 700:739 fully processed and native glycosylation 700:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 2 2 theme high-resolution 599:613 arg1 structures 615:624 high-resolution structures 599:624 high-resolution structures 599:624 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 3 3 theme native 720:725 arg1 glycosylation 727:739 fully processed and native glycosylation 700:739 fully processed and native glycosylation 700:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 0 4 theme trimer 66:71 arg1 structures 13:22 X-ray and EM structures 0:22 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer 0:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 3 5 theme trimer 688:693 arg1 structures 660:669 3.5 and 3.9 Å resolution crystal structures 627:669 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation 627:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 3 6 theme resolution 641:650 arg1 structures 660:669 3.5 and 3.9 Å resolution crystal structures 627:669 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation 627:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 3 7 theme bNAbs 877:881 arg1 epitopes 861:868 the complete epitopes 848:868 the complete epitopes of two bNAbs 848:881 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 4 8 from structures 937:946 arg1 refinement 894:903 the refinement 890:903 the refinement of the N-glycans in the crystal structures 890:946 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 4 9 theme crystal 929:935 arg1 structures 937:946 the crystal structures 925:946 the crystal structures 925:946 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 3 10 used used 833:836 arg2 shield 773:778 shield 773:778 shield 773:778 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 2 11 gly glycoprotein 403:414 arg1 Env 417:419 Env 417:419 Env 417:419 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 2 11 gly glycoprotein 403:414 arg1 glycoprotein 403:414 the trimeric envelope glycoprotein 381:414 the trimeric envelope glycoprotein (Env) 381:420 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 4 12 theme glycosylated 1011:1022 arg1 structures 1028:1037 glycosylated Env structures 1011:1037 glycosylated Env structures derived by single-particle cryo-electron microscopy 1011:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 2 13 theme envelope 394:401 arg1 Env 417:419 Env 417:419 Env 417:419 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 2 13 theme envelope 394:401 arg1 glycoprotein 403:414 the trimeric envelope glycoprotein 381:414 the trimeric envelope glycoprotein (Env) 381:420 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 4 14 theme Env 1024:1026 arg1 structures 1028:1037 glycosylated Env structures 1011:1037 glycosylated Env structures derived by single-particle cryo-electron microscopy 1011:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 2 15 theme high-mannose 470:481 arg1 forms 490:494 high-mannose glycan forms 470:494 high-mannose glycan forms only 470:499 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 2 16 theme trimeric 385:392 arg1 Env 417:419 Env 417:419 Env 417:419 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 2 16 theme trimeric 385:392 arg1 glycoprotein 403:414 the trimeric envelope glycoprotein 381:414 the trimeric envelope glycoprotein (Env) 381:420 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 0 17 theme X-ray 0:4 arg1 structures 13:22 X-ray and EM structures 0:22 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer 0:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 2 18 theme critical 304:311 arg1 mechanisms 313:322 critical mechanisms 304:322 critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env) 304:420 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 3 19 gly glycosylation 727:739 arg1 structures 660:669 3.5 and 3.9 Å resolution crystal structures 627:669 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation 627:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 4 20 theme N-glycans 912:920 arg1 refinement 894:903 the refinement 890:903 the refinement of the N-glycans in the crystal structures 890:946 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 2 21 from N-glycans 368:376 arg1 Env 417:419 Env 417:419 Env 417:419 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 2 21 from N-glycans 368:376 arg1 glycoprotein 403:414 the trimeric envelope glycoprotein 381:414 the trimeric envelope glycoprotein (Env) 381:420 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 1 22 theme structural 78:87 arg1 characterization 105:120 The structural and biochemical characterization 74:120 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) 74:174 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 22 theme structural 78:87 arg1 essential 185:193 essential 185:193 essential 185:193 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 3 23 theme crystal 652:658 arg1 structures 660:669 3.5 and 3.9 Å resolution crystal structures 627:669 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation 627:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 1 24 theme biochemical 93:103 arg1 characterization 105:120 The structural and biochemical characterization 74:120 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) 74:174 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 24 theme biochemical 93:103 arg1 essential 185:193 essential 185:193 essential 185:193 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 4 25 theme glycan 991:996 arg1 densities 998:1006 glycan densities 991:1006 glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy 991:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 4 26 gly glycosylated 1011:1022 arg1 structures 1028:1037 glycosylated Env structures 1011:1037 glycosylated Env structures derived by single-particle cryo-electron microscopy 1011:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 3 27 with structures 660:669 arg1 glycosylation 727:739 fully processed and native glycosylation 700:739 fully processed and native glycosylation 700:739 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 0 28 theme EM 10:11 arg1 structures 13:22 X-ray and EM structures 0:22 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer 0:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 4 29 from N-glycans 912:920 arg1 structures 937:946 the crystal structures 925:946 the crystal structures 925:946 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 0 30 theme glycosylated 38:49 arg1 trimer 66:71 a natively glycosylated HIV-1 envelope trimer 27:71 a natively glycosylated HIV-1 envelope trimer 27:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 3 31 theme high-mannose 783:794 arg1 N-glycans 813:821 high-mannose and complex-type N-glycans 783:821 N-glycans 813:821 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 0 32 theme HIV-1 51:55 arg1 trimer 66:71 a natively glycosylated HIV-1 envelope trimer 27:71 a natively glycosylated HIV-1 envelope trimer 27:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 3 33 theme HIV-1 678:682 arg1 trimer 688:693 the HIV-1 Env trimer 674:693 the HIV-1 Env trimer 674:693 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 4 34 from refinement 894:903 arg1 structures 937:946 the crystal structures 925:946 the crystal structures 925:946 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 2 35 theme glycans 562:568 arg1 presence 542:549 the presence 538:549 the presence of complex glycans 538:568 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 3 36 theme Env 684:686 arg1 trimer 688:693 the HIV-1 Env trimer 674:693 the HIV-1 Env trimer 674:693 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 3 37 theme N-glycans 813:821 arg1 shield 773:778 shield 773:778 shield 773:778 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 2 38 theme complex 554:560 arg1 glycans 562:568 complex glycans 554:568 complex glycans 554:568 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 1 39 theme neutralizing 133:144 arg1 antibodies 157:166 anti-HIV-1 antibodies 146:166 broadly neutralizing anti-HIV-1 antibodies (bNAbs) 125:174 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 39 theme neutralizing 133:144 arg1 bNAbs 169:173 bNAbs 169:173 bNAbs 169:173 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 40 theme potential 220:228 arg1 vaccines 230:237 potential vaccines 220:237 potential vaccines 220:237 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 0 41 theme envelope 57:64 arg1 trimer 66:71 a natively glycosylated HIV-1 envelope trimer 27:71 a natively glycosylated HIV-1 envelope trimer 27:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 4 42 theme cryo-electron 1066:1078 arg1 microscopy 1080:1089 single-particle cryo-electron microscopy 1050:1089 single-particle cryo-electron microscopy 1050:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 1 43 theme anti-HIV-1 146:155 arg1 antibodies 157:166 anti-HIV-1 antibodies 146:166 broadly neutralizing anti-HIV-1 antibodies (bNAbs) 125:174 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 43 theme anti-HIV-1 146:155 arg1 bNAbs 169:173 bNAbs 169:173 bNAbs 169:173 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 3 44 theme complete 852:859 arg1 epitopes 861:868 the complete epitopes 848:868 the complete epitopes of two bNAbs 848:881 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 28994411 1 45 theme vaccines 230:237 arg1 design 210:215 the design 206:215 the design of potential vaccines to prevent infection by HIV-1 206:267 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 2 46 theme forms 490:494 arg1 visualization 453:465 the visualization 449:465 the visualization of high-mannose glycan forms only 449:499 While these studies have revealed critical mechanisms by which bNAbs recognize and/or accommodate N-glycans on the trimeric envelope glycoprotein (Env), they have been limited to the visualization of high-mannose glycan forms only, since heterogeneity introduced from the presence of complex glycans makes it difficult to obtain high-resolution structures. 28994411 4 47 from densities 998:1006 arg1 structures 1028:1037 glycosylated Env structures 1011:1037 glycosylated Env structures derived by single-particle cryo-electron microscopy 1011:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 0 48 gly glycosylated 38:49 arg1 trimer 66:71 a natively glycosylated HIV-1 envelope trimer 27:71 a natively glycosylated HIV-1 envelope trimer 27:71 X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. 28994411 1 49 theme antibodies 157:166 arg1 characterization 105:120 The structural and biochemical characterization 74:120 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) 74:174 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 1 49 theme antibodies 157:166 arg1 essential 185:193 essential 185:193 essential 185:193 The structural and biochemical characterization of broadly neutralizing anti-HIV-1 antibodies (bNAbs) has been essential in guiding the design of potential vaccines to prevent infection by HIV-1. 28994411 4 50 theme single-particle 1050:1064 arg1 microscopy 1080:1089 single-particle cryo-electron microscopy 1050:1089 single-particle cryo-electron microscopy 1050:1089 Here, the refinement of the N-glycans in the crystal structures is discussed and comparisons are made with glycan densities in glycosylated Env structures derived by single-particle cryo-electron microscopy. 28994411 3 51 theme complex-type 800:811 arg1 N-glycans 813:821 high-mannose and complex-type N-glycans 783:821 N-glycans 813:821 3.5 and 3.9 Å resolution crystal structures of the HIV-1 Env trimer with fully processed and native glycosylation were solved, revealing a glycan shield of high-mannose and complex-type N-glycans that were used to define the complete epitopes of two bNAbs. 26610891 7 0 theme N-glycosylation 1655:1669 arg1 machineries 1671:1681 functional N-glycosylation machineries 1644:1681 functional N-glycosylation machineries 1644:1681 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 7 0 theme N-glycosylation 1655:1669 arg1 bacteria 1628:1635 deep-sea vent bacteria 1614:1635 deep-sea vent bacteria 1614:1635 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 2 1 from δ-proteobacterium 474:490 arg1 orthologues 388:398 orthologues 388:398 orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, 388:520 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 2 theme systems 652:658 arg1 subdivisions 569:580 these two subdivisions 559:580 these two subdivisions of bacteria 559:592 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 2 theme systems 652:658 arg1 source 603:608 a source 601:608 a source of further N-linked protein glycosylation systems 601:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 3 from source 753:758 arg1 subdivisions 782:793 these subdivisions 776:793 these subdivisions 776:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 1 4 theme pathogen 284:291 arg1 member 317:322 a member 315:322 a member of the ε-proteobacteria-subdivision of bacteria 315:370 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 4 theme pathogen 284:291 arg1 jejuni 307:312 the intestinal pathogen Campylobacter jejuni 269:312 the intestinal pathogen Campylobacter jejuni 269:312 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 6 5 contain contains 1536:1543 arg1 glycan 1479:1484 the N-linked glycan 1466:1484 the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum 1466:1534 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 6 5 contain contains 1536:1543 arg2 sugar 1559:1563 an acetylated sugar 1545:1563 an acetylated sugar at the reducing end 1545:1583 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 7 6 theme vent 1623:1626 arg1 machineries 1671:1681 functional N-glycosylation machineries 1644:1681 functional N-glycosylation machineries 1644:1681 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 7 6 theme vent 1623:1626 arg1 bacteria 1628:1635 deep-sea vent bacteria 1614:1635 deep-sea vent bacteria 1614:1635 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 3 7 from habitats 736:743 arg1 sequencing 674:683 Whole-genome sequencing 661:683 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, 661:794 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 5 8 theme sequon 1338:1343 arg1 specificities 1345:1357 different glycosylation sequon specificities 1314:1357 different glycosylation sequon specificities 1314:1357 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 1 9 theme selected 206:213 arg1 proteins 215:222 selected proteins 206:222 selected proteins in the periplasm 206:239 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 4 10 theme putative 955:962 arg1 ORFs 970:973 putative OTase ORFs 955:973 putative OTase ORFs 955:973 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 0 11 theme deep-sea 79:86 arg1 proteobacteria 93:106 deep-sea vent proteobacteria 79:106 deep-sea vent proteobacteria 79:106 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 2 12 theme glycosylation 638:650 arg1 systems 652:658 further N-linked protein glycosylation systems 613:658 further N-linked protein glycosylation systems 613:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 13 theme OTase 828:832 arg1 enzymes 834:840 OTase enzymes 828:840 OTase enzymes 828:840 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 6 14 theme desulfuricans 1390:1402 arg1 protein 1413:1419 a permissive D. desulfuricans acceptor protein 1374:1419 a permissive D. desulfuricans acceptor protein 1374:1419 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 2 15 theme further 613:619 arg1 systems 652:658 further N-linked protein glycosylation systems 613:658 further N-linked protein glycosylation systems 613:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 16 link N-linked 621:628 arg1 systems 652:658 further N-linked protein glycosylation systems 613:658 further N-linked protein glycosylation systems 613:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 6 17 theme D. 1387:1388 arg1 protein 1413:1419 a permissive D. desulfuricans acceptor protein 1374:1419 a permissive D. desulfuricans acceptor protein 1374:1419 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 3 18 theme adjacent 857:864 arg1 glycosyltransferases 866:885 associated adjacent glycosyltransferases 846:885 associated adjacent glycosyltransferases 846:885 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 6 19 theme N-linked 1470:1477 arg1 glycan 1479:1484 the N-linked glycan 1466:1484 the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum 1466:1534 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 4 20 theme vent 993:996 arg1 species 998:1004 the deep-sea vent species 980:1004 the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli 980:1107 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 20 theme vent 993:996 arg1 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 20 theme vent 993:996 arg1 Sulfurovum 1031:1040 Sulfurovum 1031:1040 Sulfurovum 1031:1040 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 20 theme vent 993:996 arg1 Deferribacter 1061:1073 Deferribacter 1061:1073 Deferribacter 1061:1073 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 3 21 theme Whole-genome 661:672 arg1 sequencing 674:683 Whole-genome sequencing 661:683 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, 661:794 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 2 22 theme protein 630:636 arg1 systems 652:658 further N-linked protein glycosylation systems 613:658 further N-linked protein glycosylation systems 613:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 23 theme δ-proteobacteria 700:715 arg1 sequencing 674:683 Whole-genome sequencing 661:683 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, 661:794 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 1 24 from proteins 215:222 arg1 periplasm 231:239 the periplasm 227:239 the periplasm 227:239 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 3 25 theme similar 887:893 arg1 enzymes 834:840 OTase enzymes 828:840 OTase enzymes 828:840 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 4 26 dep species 998:1004 arg1 species 998:1004 the deep-sea vent species 980:1004 the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli 980:1107 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 26 dep species 998:1004 arg1 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 26 dep species 998:1004 arg1 Sulfurovum 1031:1040 Sulfurovum 1031:1040 Sulfurovum 1031:1040 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 26 dep species 998:1004 arg1 Deferribacter 1061:1073 Deferribacter 1061:1073 Deferribacter 1061:1073 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 5 27 theme relaxed 1230:1236 arg1 specificity 1245:1255 relaxed glycan specificity 1230:1255 relaxed glycan specificity 1230:1255 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 3 28 theme ε- 693:694 arg1 sequencing 674:683 Whole-genome sequencing 661:683 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, 661:794 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 29 theme glycosylation 921:933 arg1 locus 935:939 the C. jejuni N-linked glycosylation locus 898:939 the C. jejuni N-linked glycosylation locus 898:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 0 30 theme vent 88:91 arg1 proteobacteria 93:106 deep-sea vent proteobacteria 79:106 deep-sea vent proteobacteria 79:106 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 5 31 contain possess 1222:1228 arg2 specificity 1245:1255 relaxed glycan specificity 1230:1255 relaxed glycan specificity 1230:1255 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 5 31 contain possess 1222:1228 arg1 enzymes 1200:1206 The enzymes 1196:1206 The enzymes 1196:1206 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 3 32 link N-linked 912:919 arg1 locus 935:939 the C. jejuni N-linked glycosylation locus 898:939 the C. jejuni N-linked glycosylation locus 898:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 33 theme jejuni 905:910 arg1 locus 935:939 the C. jejuni N-linked glycosylation locus 898:939 the C. jejuni N-linked glycosylation locus 898:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 4 34 dep Sulfurovum 1031:1040 arg1 lithotrophicum 1042:1055 Sulfurovum lithotrophicum 1031:1055 Sulfurovum lithotrophicum 1031:1055 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 7 35 theme OTase 1742:1746 arg1 enzymes 1748:1754 biotechnologically important OTase enzymes 1713:1754 biotechnologically important OTase enzymes 1713:1754 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 2 36 theme bacteria 585:592 arg1 subdivisions 569:580 these two subdivisions 559:580 these two subdivisions of bacteria 559:592 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 36 theme bacteria 585:592 arg1 source 603:608 a source 601:608 a source of further N-linked protein glycosylation systems 601:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 0 37 theme transferase 48:58 arg1 enzymes 60:66 N-linking oligosaccharyl transferase enzymes 23:66 N-linking oligosaccharyl transferase enzymes 23:66 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 6 38 theme acetylated 1548:1557 arg1 sugar 1559:1563 an acetylated sugar 1545:1563 an acetylated sugar at the reducing end 1545:1583 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 0 39 theme Functional 0:9 arg1 analysis 11:18 Functional analysis 0:18 Functional analysis of N-linking oligosaccharyl transferase enzymes 0:66 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 1 40 theme Bacterial 109:117 arg1 transferases 144:155 Bacterial N-linking oligosaccharyl transferases 109:155 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) 109:171 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 40 theme Bacterial 109:117 arg1 enzymes 164:170 OTase enzymes 158:170 OTase enzymes 158:170 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 41 link lipid-linked 182:193 arg1 glycans 195:201 lipid-linked glycans 182:201 lipid-linked glycans 182:201 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 5 42 theme different 1314:1322 arg1 specificities 1345:1357 different glycosylation sequon specificities 1314:1357 different glycosylation sequon specificities 1314:1357 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 1 43 theme oligosaccharyl 129:142 arg1 transferases 144:155 Bacterial N-linking oligosaccharyl transferases 109:155 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) 109:171 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 43 theme oligosaccharyl 129:142 arg1 enzymes 164:170 OTase enzymes 158:170 OTase enzymes 158:170 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 0 44 theme oligosaccharyl 33:46 arg1 enzymes 60:66 N-linking oligosaccharyl transferase enzymes 23:66 N-linking oligosaccharyl transferase enzymes 23:66 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 7 45 theme potential 1693:1701 arg1 source 1703:1708 a potential source 1691:1708 a potential source of biotechnologically important OTase enzymes 1691:1754 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 3 46 theme deep-sea 722:729 arg1 habitats 736:743 deep-sea vent habitats 722:743 deep-sea vent habitats 722:743 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 46 theme deep-sea 722:729 arg1 source 753:758 a rich source 746:758 a rich source of species from these subdivisions 746:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 0 47 theme enzymes 60:66 arg1 analysis 11:18 Functional analysis 0:18 Functional analysis of N-linking oligosaccharyl transferase enzymes 0:66 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 5 48 theme glycan 1279:1284 arg1 structures 1286:1295 diverse glycan structures 1271:1295 diverse glycan structures 1271:1295 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 2 49 from species 460:466 arg1 orthologues 388:398 orthologues 388:398 orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, 388:520 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 50 theme ε-proteobacterial 411:427 arg1 Campylobacter 429:441 other ε-proteobacterial Campylobacter and Helicobacter species 405:466 Campylobacter 429:441 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 7 51 theme functional 1644:1653 arg1 machineries 1671:1681 functional N-glycosylation machineries 1644:1681 functional N-glycosylation machineries 1644:1681 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 7 51 theme functional 1644:1653 arg1 bacteria 1628:1635 deep-sea vent bacteria 1614:1635 deep-sea vent bacteria 1614:1635 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 5 52 theme diverse 1271:1277 arg1 structures 1286:1295 diverse glycan structures 1271:1295 diverse glycan structures 1271:1295 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 1 53 theme intestinal 273:282 arg1 member 317:322 a member 315:322 a member of the ε-proteobacteria-subdivision of bacteria 315:370 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 53 theme intestinal 273:282 arg1 jejuni 307:312 the intestinal pathogen Campylobacter jejuni 269:312 the intestinal pathogen Campylobacter jejuni 269:312 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 4 54 theme OTase 964:968 arg1 ORFs 970:973 putative OTase ORFs 955:973 putative OTase ORFs 955:973 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 3 55 theme species 763:769 arg1 habitats 736:743 deep-sea vent habitats 722:743 deep-sea vent habitats 722:743 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 55 theme species 763:769 arg1 source 753:758 a rich source 746:758 a rich source of species from these subdivisions 746:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 1 56 theme Campylobacter 293:305 arg1 member 317:322 a member 315:322 a member of the ε-proteobacteria-subdivision of bacteria 315:370 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 56 theme Campylobacter 293:305 arg1 jejuni 307:312 the intestinal pathogen Campylobacter jejuni 269:312 the intestinal pathogen Campylobacter jejuni 269:312 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 7 57 theme deep-sea 1614:1621 arg1 machineries 1671:1681 functional N-glycosylation machineries 1644:1681 functional N-glycosylation machineries 1644:1681 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 7 57 theme deep-sea 1614:1621 arg1 bacteria 1628:1635 deep-sea vent bacteria 1614:1635 deep-sea vent bacteria 1614:1635 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 3 58 theme putative 805:812 arg1 ORFs 814:817 putative ORFs 805:817 putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus 805:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 59 from subdivisions 782:793 arg1 habitats 736:743 deep-sea vent habitats 722:743 deep-sea vent habitats 722:743 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 59 from subdivisions 782:793 arg1 species 763:769 species 763:769 species from these subdivisions 763:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 59 from subdivisions 782:793 arg1 source 753:758 a rich source 746:758 a rich source of species from these subdivisions 746:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 6 60 theme acceptor 1404:1411 arg1 protein 1413:1419 a permissive D. desulfuricans acceptor protein 1374:1419 a permissive D. desulfuricans acceptor protein 1374:1419 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 4 61 dep Deferribacter 1061:1073 arg1 desulfuricans 1075:1087 Deferribacter desulfuricans 1061:1087 Deferribacter desulfuricans 1061:1087 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 6 62 from end 1581:1583 arg1 sugar 1559:1563 an acetylated sugar 1545:1563 an acetylated sugar at the reducing end 1545:1583 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 6 63 theme permissive 1376:1385 arg1 protein 1413:1419 a permissive D. desulfuricans acceptor protein 1374:1419 a permissive D. desulfuricans acceptor protein 1374:1419 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 4 64 theme deep-sea 984:991 arg1 species 998:1004 the deep-sea vent species 980:1004 the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli 980:1107 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 64 theme deep-sea 984:991 arg1 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 Nitratiruptor 1006:1018 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 64 theme deep-sea 984:991 arg1 Sulfurovum 1031:1040 Sulfurovum 1031:1040 Sulfurovum 1031:1040 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 4 64 theme deep-sea 984:991 arg1 Deferribacter 1061:1073 Deferribacter 1061:1073 Deferribacter 1061:1073 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 3 65 theme associated 846:855 arg1 glycosyltransferases 866:885 associated adjacent glycosyltransferases 846:885 associated adjacent glycosyltransferases 846:885 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 4 66 from species 998:1004 arg1 coli 1104:1107 Escherichia coli 1092:1107 Escherichia coli 1092:1107 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 1 67 theme OTase 158:162 arg1 transferases 144:155 Bacterial N-linking oligosaccharyl transferases 109:155 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) 109:171 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 67 theme OTase 158:162 arg1 enzymes 164:170 OTase enzymes 158:170 OTase enzymes 158:170 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 2 68 theme N-linked 621:628 arg1 systems 652:658 further N-linked protein glycosylation systems 613:658 further N-linked protein glycosylation systems 613:658 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 69 theme C. 902:903 arg1 jejuni 905:910 C. jejuni 902:910 the C. jejuni N-linked glycosylation locus 898:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 4 70 dep Nitratiruptor 1006:1018 arg1 tergarcus 1020:1028 Nitratiruptor tergarcus 1006:1028 Nitratiruptor tergarcus 1006:1028 We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. 26610891 5 71 theme glycan 1238:1243 arg1 specificity 1245:1255 relaxed glycan specificity 1230:1255 relaxed glycan specificity 1230:1255 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 1 72 theme lipid-linked 182:193 arg1 glycans 195:201 lipid-linked glycans 182:201 lipid-linked glycans 182:201 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 7 73 theme enzymes 1748:1754 arg1 source 1703:1708 a potential source 1691:1708 a potential source of biotechnologically important OTase enzymes 1691:1754 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 7 74 theme important 1732:1740 arg1 enzymes 1748:1754 biotechnologically important OTase enzymes 1713:1754 biotechnologically important OTase enzymes 1713:1754 This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes. 26610891 3 75 theme N-linked 912:919 arg1 locus 935:939 the C. jejuni N-linked glycosylation locus 898:939 the C. jejuni N-linked glycosylation locus 898:939 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 2 76 theme Helicobacter 447:458 arg1 species 460:466 other ε-proteobacterial Campylobacter and Helicobacter species 405:466 species 460:466 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 0 77 theme N-linking 23:31 arg1 enzymes 60:66 N-linking oligosaccharyl transferase enzymes 23:66 N-linking oligosaccharyl transferase enzymes 23:66 Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria. 26610891 1 78 theme N-linking 119:127 arg1 transferases 144:155 Bacterial N-linking oligosaccharyl transferases 109:155 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) 109:171 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 78 theme N-linking 119:127 arg1 enzymes 164:170 OTase enzymes 158:170 OTase enzymes 158:170 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 2 79 theme Desulfovibrio 493:505 arg1 δ-proteobacterium 474:490 a δ-proteobacterium 472:490 a δ-proteobacterium 472:490 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 2 79 theme Desulfovibrio 493:505 arg1 desulfuricans 507:519 Desulfovibrio desulfuricans 493:519 Desulfovibrio desulfuricans 493:519 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 80 theme vent 731:734 arg1 habitats 736:743 deep-sea vent habitats 722:743 deep-sea vent habitats 722:743 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 80 theme vent 731:734 arg1 source 753:758 a rich source 746:758 a rich source of species from these subdivisions 746:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 6 81 theme reducing 1572:1579 arg1 end 1581:1583 the reducing end 1568:1583 the reducing end 1568:1583 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 5 82 theme glycosylation 1324:1336 arg1 specificities 1345:1357 different glycosylation sequon specificities 1314:1357 different glycosylation sequon specificities 1314:1357 The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. 26610891 1 83 theme ε-proteobacteria-subdivision 331:358 arg1 member 317:322 a member 315:322 a member of the ε-proteobacteria-subdivision of bacteria 315:370 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 1 83 theme ε-proteobacteria-subdivision 331:358 arg1 jejuni 307:312 the intestinal pathogen Campylobacter jejuni 269:312 the intestinal pathogen Campylobacter jejuni 269:312 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 2 84 from Campylobacter 429:441 arg1 orthologues 388:398 orthologues 388:398 orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, 388:520 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 26610891 3 85 theme rich 748:751 arg1 habitats 736:743 deep-sea vent habitats 722:743 deep-sea vent habitats 722:743 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 3 85 theme rich 748:751 arg1 source 753:758 a rich source 746:758 a rich source of species from these subdivisions 746:793 Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. 26610891 1 86 theme bacteria 363:370 arg1 ε-proteobacteria-subdivision 331:358 the ε-proteobacteria-subdivision 327:358 the ε-proteobacteria-subdivision of bacteria 327:370 Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. 26610891 6 87 link N-linked 1470:1477 arg1 glycan 1479:1484 the N-linked glycan 1466:1484 the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum 1466:1534 Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. 26610891 2 88 theme other 405:409 arg1 Campylobacter 429:441 other ε-proteobacterial Campylobacter and Helicobacter species 405:466 Campylobacter 429:441 More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. 28199092 7 0 theme chromatography 1768:1781 arg1 analysis 1791:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis 1721:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 6 1 with glycopeptides 1386:1398 arg1 classes 1415:1421 different classes 1405:1421 different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1405:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 1 2 theme glycosylation 285:297 arg1 information 299:309 site-specific glycosylation information 271:309 site-specific glycosylation information 271:309 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 2 3 attach present 407:413 arg2 species 399:405 the individual glycan species 377:405 the individual glycan species present at a particular site 377:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 2 3 attach present 407:413 arg1 site 431:434 a particular site 418:434 a particular site 418:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 7 4 theme interaction 1749:1759 arg1 HILIC 1784:1788 HILIC 1784:1788 HILIC 1784:1788 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 4 theme interaction 1749:1759 arg1 chromatography 1768:1781 the traditional hydrophilic interaction liquid chromatography 1721:1781 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 5 theme glycan 1010:1015 arg1 present 1031:1037 N-linked glycan heterogeneity present 1001:1037 N-linked glycan heterogeneity present 1001:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 8 6 from glycoform 1905:1913 arg1 impact 1889:1894 no significant impact 1874:1894 no significant impact from the glycoform on the ionization properties of the glycopeptide 1874:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 3 7 theme multiattribute 607:620 arg1 MAM 630:632 MAM 630:632 MAM 630:632 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 7 theme multiattribute 607:620 arg1 method 622:627 a multiattribute method 605:627 a multiattribute method (MAM) 605:633 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 4 8 attach present 1121:1127 arg1 chain 1142:1146 the heavy chain 1132:1146 the heavy chain (HC) 1132:1151 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 8 attach present 1121:1127 arg1 HC 1149:1150 HC 1149:1150 HC 1149:1150 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 8 attach present 1121:1127 arg2 region 1162:1167 one present on the heavy chain (HC) variable region 1117:1167 one present on the heavy chain (HC) variable region (Fab) 1117:1173 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 8 attach present 1121:1127 arg2 Fab 1170:1172 Fab 1170:1172 Fab 1170:1172 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 7 9 theme traditional 1725:1735 arg1 HILIC 1784:1788 HILIC 1784:1788 HILIC 1784:1788 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 9 theme traditional 1725:1735 arg1 chromatography 1768:1781 the traditional hydrophilic interaction liquid chromatography 1721:1781 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 10 theme distinct 1078:1085 arg1 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 9 11 theme glycans 2129:2135 arg1 profiling 2107:2115 profiling 2107:2115 profiling of N-linked glycans on immunoglobulins 2107:2154 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 4 12 theme glycosylation 1096:1108 arg1 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 13 link N-linked 1001:1008 arg1 present 1031:1037 N-linked glycan heterogeneity present 1001:1037 N-linked glycan heterogeneity present 1001:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 2 14 theme glycosylation 356:368 arg1 sites 370:374 multiple glycosylation sites 347:374 multiple glycosylation sites 347:374 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 6 15 theme variants 1444:1451 arg1 classes 1415:1421 different classes 1405:1421 different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1405:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 3 16 dep quantify 646:653 arg1 including 697:705 including 697:705 including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1) 697:897 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 7 17 from sites 1675:1679 arg1 species 1619:1625 individual glycan species 1601:1625 individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1601:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 17 from sites 1675:1679 arg1 quantification 1583:1596 MAM quantification 1579:1596 MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1579:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 18 theme mAb 1050:1052 arg1 molecule 1054:1061 an IgG1 mAb molecule 1042:1061 an IgG1 mAb molecule containing two distinct N-linked glycosylation sites 1042:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 6 19 theme glycan 1426:1431 arg1 variants 1444:1451 glycan structural variants 1426:1451 glycan structural variants 1426:1451 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 6 19 theme glycan 1426:1431 arg1 mannose 1469:1475 high mannose 1464:1475 high mannose 1464:1475 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 1 20 theme techniques 232:241 arg1 challenge 192:200 A significant challenge 178:200 A significant challenge of traditional glycan mapping techniques 178:241 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 4 21 attach present 1189:1195 arg2 region 1226:1231 the other present on the conserved HC constant region 1179:1231 the other present on the conserved HC constant region (Fc) 1179:1236 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 21 attach present 1189:1195 arg1 HC 1214:1215 the conserved HC 1200:1215 the conserved HC 1200:1215 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 21 attach present 1189:1195 arg2 Fc 1234:1235 Fc 1234:1235 Fc 1234:1235 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 0 22 theme N-Linked 148:155 arg1 Sites 171:175 Two N-Linked Glycosylation Sites 144:175 Two N-Linked Glycosylation Sites 144:175 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 6 23 theme galactosylated 1503:1516 arg1 species 1518:1524 terminal galactosylated species 1494:1524 terminal galactosylated species 1494:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 6 23 theme galactosylated 1503:1516 arg1 mannose 1469:1475 high mannose 1464:1475 high mannose 1464:1475 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 4 24 dep describe 917:924 arg1 Fc 1234:1235 Fc 1234:1235 Fc 1234:1235 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 24 dep describe 917:924 arg1 Fab 1170:1172 Fab 1170:1172 Fab 1170:1172 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 24 dep describe 917:924 arg1 region 1226:1231 the other present on the conserved HC constant region 1179:1231 the other present on the conserved HC constant region (Fc) 1179:1236 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 24 dep describe 917:924 arg1 region 1162:1167 one present on the heavy chain (HC) variable region 1117:1167 one present on the heavy chain (HC) variable region (Fab) 1117:1173 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 7 25 theme N-Linked 1652:1659 arg1 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 0 26 theme Monitoring 62:71 arg1 Heterogeneity 94:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 4 27 theme specific 974:981 arg1 quantification 983:996 site specific quantification 969:996 site specific quantification of N-linked glycan heterogeneity present 969:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 6 28 theme sialylated 1478:1487 arg1 classes 1415:1421 different classes 1405:1421 different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1405:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 4 29 theme present 1121:1127 arg1 Fab 1170:1172 Fab 1170:1172 Fab 1170:1172 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 29 theme present 1121:1127 arg1 region 1162:1167 one present on the heavy chain (HC) variable region 1117:1167 one present on the heavy chain (HC) variable region (Fab) 1117:1173 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 0 30 theme Glycan 87:92 arg1 Heterogeneity 94:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 4 31 gly glycosylation 1096:1108 arg2 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 31 gly glycosylation 1096:1108 arg2 two 1074:1076 two 1074:1076 two 1074:1076 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 1 32 theme significant 180:190 arg1 challenge 192:200 A significant challenge 178:200 A significant challenge of traditional glycan mapping techniques 178:241 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 0 33 from Application 0:10 arg1 Antibody 124:131 a Monoclonal Antibody 111:131 a Monoclonal Antibody 111:131 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 8 34 theme glycopeptide 1951:1962 arg1 properties 1933:1942 the ionization properties 1918:1942 the ionization properties of the glycopeptide 1918:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 9 35 dep quantitative 2056:2067 arg1 site-specific 2070:2082 site-specific 2070:2082 site-specific 2070:2082 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 0 36 theme Monoclonal 113:122 arg1 Antibody 124:131 a Monoclonal Antibody 111:131 a Monoclonal Antibody 111:131 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 6 37 gly glycopeptides 1386:1398 arg2 glycopeptides 1386:1398 the glycopeptides 1382:1398 the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1382:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 7 38 theme glycan 1612:1617 arg1 species 1619:1625 individual glycan species 1601:1625 individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1601:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 2 39 theme other 495:499 arg1 sites 515:519 the other glycosylation sites 491:519 the other glycosylation sites on the molecule 491:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 7 40 dep Fab 1641:1643 arg1 the 1637:1639 the 1637:1639 the 1637:1639 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 2 41 attach present 480:486 arg2 species 472:478 those species 466:478 those species present at the other glycosylation sites on the molecule 466:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 2 41 attach present 480:486 arg1 sites 515:519 the other glycosylation sites 491:519 the other glycosylation sites on the molecule 491:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 4 42 theme variable 1153:1160 arg1 Fab 1170:1172 Fab 1170:1172 Fab 1170:1172 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 42 theme variable 1153:1160 arg1 region 1162:1167 one present on the heavy chain (HC) variable region 1117:1167 one present on the heavy chain (HC) variable region (Fab) 1117:1173 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 7 43 theme MAM 1579:1581 arg1 quantification 1583:1596 MAM quantification 1579:1596 MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1579:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 44 theme MAM 948:950 arg1 application 930:940 the application 926:940 the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites 926:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 9 45 theme suitable 2024:2031 arg1 approach 2033:2040 a suitable approach 2022:2040 a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins 2022:2154 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 9 45 theme suitable 2024:2031 arg1 method 2012:2017 the MAM method 2004:2017 the MAM method 2004:2017 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 0 46 theme LC-MS 30:34 arg1 Method 51:56 a Quantitative LC-MS Multiattribute Method 15:56 a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity 15:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 2 47 contain containing 336:345 arg2 sites 370:374 multiple glycosylation sites 347:374 multiple glycosylation sites 347:374 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 2 47 contain containing 336:345 arg1 proteins 327:334 proteins 327:334 proteins containing multiple glycosylation sites 327:374 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 9 48 from profiling 2107:2115 arg1 immunoglobulins 2140:2154 immunoglobulins 2140:2154 immunoglobulins 2140:2154 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 2 49 from site 431:434 arg1 present 407:413 present 407:413 present 407:413 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 8 50 theme ionization 1922:1931 arg1 properties 1933:1942 the ionization properties 1918:1942 the ionization properties of the glycopeptide 1918:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 2 51 theme glycan 392:397 arg1 species 399:405 the individual glycan species 377:405 the individual glycan species present at a particular site 377:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 0 52 theme Method 51:56 arg1 Application 0:10 Application 0:10 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody 0:131 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 7 53 theme labeled 1844:1850 arg1 glycans 1852:1858 fluorescently labeled glycans 1830:1858 fluorescently labeled glycans 1830:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 3 54 theme post-translational 664:681 arg1 modifications 683:695 multiple post-translational modifications 655:695 multiple post-translational modifications 655:695 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 9 55 theme quantitative 2056:2067 arg1 information 2091:2101 quantitative, site-specific glycan information 2056:2101 quantitative, site-specific glycan information 2056:2101 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 6 56 theme different 1405:1413 arg1 classes 1415:1421 different classes 1405:1421 different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1405:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 3 57 dep deamidation 707:717 arg1 MAbs 864:867 Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 775:882 Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1 775:896 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 7 58 gly glycosylation 1661:1673 arg2 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 59 theme IgG1 1045:1048 arg1 molecule 1054:1061 an IgG1 mAb molecule 1042:1061 an IgG1 mAb molecule containing two distinct N-linked glycosylation sites 1042:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 8 60 from impact 1889:1894 arg1 properties 1933:1942 the ionization properties 1918:1942 the ionization properties of the glycopeptide 1918:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 2 61 gly glycosylation 356:368 arg2 sites 370:374 multiple glycosylation sites 347:374 multiple glycosylation sites 347:374 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 4 62 theme present 1189:1195 arg1 Fc 1234:1235 Fc 1234:1235 Fc 1234:1235 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 62 theme present 1189:1195 arg1 region 1226:1231 the other present on the conserved HC constant region 1179:1231 the other present on the conserved HC constant region (Fc) 1179:1236 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 5 63 theme mapping 1256:1262 arg1 method 1264:1269 a peptide mapping method 1246:1269 a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest 1246:1350 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 5 63 theme mapping 1256:1262 arg1 MAM 1239:1241 MAM 1239:1241 MAM 1239:1241 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 6 64 theme glycopeptides 1386:1398 arg1 properties 1368:1377 The ionization properties 1353:1377 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1353:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 7 65 theme liquid 1761:1766 arg1 HILIC 1784:1788 HILIC 1784:1788 HILIC 1784:1788 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 65 theme liquid 1761:1766 arg1 chromatography 1768:1781 the traditional hydrophilic interaction liquid chromatography 1721:1781 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 66 theme N-linked 1001:1008 arg1 present 1031:1037 N-linked glycan heterogeneity present 1001:1037 N-linked glycan heterogeneity present 1001:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 2 67 theme present 407:413 arg1 species 399:405 the individual glycan species 377:405 the individual glycan species present at a particular site 377:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 1 68 theme site-specific 271:283 arg1 information 299:309 site-specific glycosylation information 271:309 site-specific glycosylation information 271:309 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 7 69 theme hydrophilic 1737:1747 arg1 HILIC 1784:1788 HILIC 1784:1788 HILIC 1784:1788 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 69 theme hydrophilic 1737:1747 arg1 chromatography 1768:1781 the traditional hydrophilic interaction liquid chromatography 1721:1781 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 70 theme heterogeneity 1017:1029 arg1 present 1031:1037 N-linked glycan heterogeneity present 1001:1037 N-linked glycan heterogeneity present 1001:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 5 71 theme interest 1343:1350 arg1 peptides 1331:1338 specific peptides 1322:1338 specific peptides of interest 1322:1350 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 6 72 theme ionization 1357:1366 arg1 properties 1368:1377 The ionization properties 1353:1377 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1353:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 4 73 theme N-linked 1087:1094 arg1 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 9 74 theme N-linked 2120:2127 arg1 glycans 2129:2135 N-linked glycans 2120:2135 N-linked glycans 2120:2135 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 6 75 theme high 1464:1467 arg1 mannose 1469:1475 high mannose 1464:1475 high mannose 1464:1475 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 3 76 theme S. 786:787 arg1 MAbs 864:867 Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 775:882 Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1 775:896 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 9 77 link N-linked 2120:2127 arg1 glycans 2129:2135 N-linked glycans 2120:2135 N-linked glycans 2120:2135 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 1 78 theme traditional 205:215 arg1 techniques 232:241 traditional glycan mapping techniques 205:241 traditional glycan mapping techniques 205:241 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 5 79 theme specific 1322:1329 arg1 peptides 1331:1338 specific peptides 1322:1338 specific peptides of interest 1322:1350 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 8 80 theme significant 1877:1887 arg1 impact 1889:1894 no significant impact 1874:1894 no significant impact from the glycoform on the ionization properties of the glycopeptide 1874:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 2 81 theme multiple 347:354 arg1 sites 370:374 multiple glycosylation sites 347:374 multiple glycosylation sites 347:374 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 6 82 theme structural 1433:1442 arg1 variants 1444:1451 glycan structural variants 1426:1451 glycan structural variants 1426:1451 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 6 82 theme structural 1433:1442 arg1 mannose 1469:1475 high mannose 1464:1475 high mannose 1464:1475 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 1 83 theme mapping 224:230 arg1 techniques 232:241 traditional glycan mapping techniques 205:241 traditional glycan mapping techniques 205:241 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 3 84 theme glycoform 556:564 arg1 Quantification 538:551 Quantification 538:551 Quantification of glycoform 538:564 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 0 85 theme Glycosylation 157:169 arg1 Sites 171:175 Two N-Linked Glycosylation Sites 144:175 Two N-Linked Glycosylation Sites 144:175 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 1 86 theme glycan 217:222 arg1 techniques 232:241 traditional glycan mapping techniques 205:241 traditional glycan mapping techniques 205:241 A significant challenge of traditional glycan mapping techniques is that they do not provide site-specific glycosylation information. 28199092 7 87 from quantification 1583:1596 arg1 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 88 theme heavy 1136:1140 arg1 HC 1149:1150 HC 1149:1150 HC 1149:1150 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 88 theme heavy 1136:1140 arg1 chain 1142:1146 the heavy chain 1132:1146 the heavy chain (HC) 1132:1151 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 6 89 theme species 1518:1524 arg1 classes 1415:1421 different classes 1405:1421 different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species 1405:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 6 90 theme terminal 1494:1501 arg1 species 1518:1524 terminal galactosylated species 1494:1524 terminal galactosylated species 1494:1524 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 6 90 theme terminal 1494:1501 arg1 mannose 1469:1475 high mannose 1464:1475 high mannose 1464:1475 The ionization properties of the glycopeptides with different classes of glycan structural variants, including high mannose, sialylated, and terminal galactosylated species were studied in detail. 28199092 8 91 gly glycopeptide 1951:1962 arg2 glycopeptide 1951:1962 the glycopeptide 1947:1962 the glycopeptide 1947:1962 Furthermore, no significant impact from the glycoform on the ionization properties of the glycopeptide is observed. 28199092 0 92 theme Site-Specific 73:85 arg1 Heterogeneity 94:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Monitoring Site-Specific Glycan Heterogeneity 62:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 3 93 theme multiple 655:662 arg1 modifications 683:695 multiple post-translational modifications 655:695 multiple post-translational modifications 655:695 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 7 94 theme glycosylation 1661:1673 arg1 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 95 theme Fc 1649:1650 arg1 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 96 theme Fab 1641:1643 arg1 sites 1675:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 both the Fab and Fc N-Linked glycosylation sites 1632:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 2 97 from sites 515:519 arg1 molecule 528:535 the molecule 524:535 the molecule 524:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 2 97 from sites 515:519 arg1 present 480:486 present 480:486 present 480:486 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 3 98 dep MAbs 864:867 arg1 890 886:888 890 886:888 890 886:888 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 98 dep MAbs 864:867 arg1 881 880:882 881 880:882 881 880:882 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 98 dep MAbs 864:867 arg1 2015 869:872 2015 869:872 2015 869:872 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 98 dep MAbs 864:867 arg1 ref 892:894 ref 1 892:896 Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1 775:896 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 7 99 theme species 1619:1625 arg1 quantification 1583:1596 MAM quantification 1579:1596 MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1579:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 7 100 with consistent 1684:1693 arg1 quantification 1700:1713 quantification 1700:1713 quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1700:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 2 101 theme particular 420:429 arg1 site 431:434 a particular site 418:434 a particular site 418:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 0 102 theme Quantitative 17:28 arg1 Method 51:56 a Quantitative LC-MS Multiattribute Method 15:56 a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity 15:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 4 103 contain containing 1063:1072 arg2 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 103 contain containing 1063:1072 arg1 molecule 1054:1061 an IgG1 mAb molecule 1042:1061 an IgG1 mAb molecule containing two distinct N-linked glycosylation sites 1042:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 7 104 theme individual 1601:1610 arg1 species 1619:1625 individual glycan species 1601:1625 individual glycan species from both the Fab and Fc N-Linked glycosylation sites 1601:1679 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 2 105 theme present 480:486 arg1 species 472:478 those species 466:478 those species present at the other glycosylation sites on the molecule 466:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 4 106 theme conserved 1204:1212 arg1 HC 1214:1215 the conserved HC 1200:1215 the conserved HC 1200:1215 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 0 107 theme Multiattribute 36:49 arg1 Method 51:56 a Quantitative LC-MS Multiattribute Method 15:56 a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity 15:106 Application of a Quantitative LC-MS Multiattribute Method for Monitoring Site-Specific Glycan Heterogeneity on a Monoclonal Antibody Containing Two N-Linked Glycosylation Sites. 28199092 7 108 theme glycans 1852:1858 arg1 analysis 1791:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis 1721:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 109 theme constant 1217:1224 arg1 Fc 1234:1235 Fc 1234:1235 Fc 1234:1235 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 109 theme constant 1217:1224 arg1 region 1226:1231 the other present on the conserved HC constant region 1179:1231 the other present on the conserved HC constant region (Fc) 1179:1236 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 9 110 theme MAM 2008:2010 arg1 approach 2033:2040 a suitable approach 2022:2040 a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins 2022:2154 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 9 110 theme MAM 2008:2010 arg1 method 2012:2017 the MAM method 2004:2017 the MAM method 2004:2017 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28199092 5 111 theme mass 1281:1284 arg1 spectrometry 1286:1297 mass spectrometry 1281:1297 mass spectrometry 1281:1297 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 3 112 dep S. 786:787 arg1 P. 836:837 P. 836:837 P. 836:837 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 112 dep S. 786:787 arg1 A. 861:862 A. 861:862 A. 861:862 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 112 dep S. 786:787 arg1 B. 822:823 B. 822:823 B. 822:823 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 112 dep S. 786:787 arg1 R. 848:849 R. 848:849 R. 848:849 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 112 dep S. 786:787 arg1 N. 803:804 N. 803:804 N. 803:804 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 3 112 dep S. 786:787 arg1 S. 806:807 S. 806:807 S. 806:807 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 2 113 gly glycosylation 501:513 arg2 sites 515:519 the other glycosylation sites 491:519 the other glycosylation sites on the molecule 491:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 2 114 theme glycosylation 501:513 arg1 sites 515:519 the other glycosylation sites 491:519 the other glycosylation sites on the molecule 491:535 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 4 115 theme other 1183:1187 arg1 Fc 1234:1235 Fc 1234:1235 Fc 1234:1235 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 115 theme other 1183:1187 arg1 region 1226:1231 the other present on the conserved HC constant region 1179:1231 the other present on the conserved HC constant region (Fc) 1179:1236 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 4 116 link N-linked 1087:1094 arg1 sites 1110:1114 two distinct N-linked glycosylation sites 1074:1114 two distinct N-linked glycosylation sites 1074:1114 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 2 117 theme individual 381:390 arg1 species 399:405 the individual glycan species 377:405 the individual glycan species present at a particular site 377:434 Therefore, for proteins containing multiple glycosylation sites, the individual glycan species present at a particular site cannot be differentiated from those species present at the other glycosylation sites on the molecule. 28199092 7 118 theme released 1817:1824 arg1 analysis 1791:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis 1721:1798 the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans 1721:1858 Our results demonstrate that MAM quantification of individual glycan species from both the Fab and Fc N-Linked glycosylation sites is consistent with quantification using the traditional hydrophilic interaction liquid chromatography (HILIC) analysis of enzymatically released and fluorescently labeled glycans. 28199092 4 119 theme present 1031:1037 arg1 quantification 983:996 site specific quantification 969:996 site specific quantification of N-linked glycan heterogeneity present 969:1037 In this paper we describe the application of an MAM based method for site specific quantification of N-linked glycan heterogeneity present on an IgG1 mAb molecule containing two distinct N-linked glycosylation sites: one present on the heavy chain (HC) variable region (Fab) and the other present on the conserved HC constant region (Fc). 28199092 3 120 theme glycosylation 731:743 arg1 variants 745:752 glycosylation variants 731:752 glycosylation variants 731:752 Quantification of glycoform has previously been demonstrated using a multiattribute method (MAM), which can quantify multiple post-translational modifications including deamidation, oxidation, glycosylation variants, and fragmentation ( Rogers, R. S.; Nightlinger, N. S.; Livingston, B.; Campbell, P.; Bailey, R.; Balland, A. MAbs 2015 , 7 , 881 - 890 ; ref 1). 28199092 5 121 theme peptide 1248:1254 arg1 method 1264:1269 a peptide mapping method 1246:1269 a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest 1246:1350 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 5 121 theme peptide 1248:1254 arg1 MAM 1239:1241 MAM 1239:1241 MAM 1239:1241 MAM is a peptide mapping method utilizing mass spectrometry to detect and quantify specific peptides of interest. 28199092 9 122 theme glycan 2084:2089 arg1 information 2091:2101 quantitative, site-specific glycan information 2056:2101 quantitative, site-specific glycan information 2056:2101 Our work demonstrates that the MAM method is a suitable approach for providing quantitative, site-specific glycan information for profiling of N-linked glycans on immunoglobulins. 28533470 0 0 theme Epithelial 83:92 arg1 Sites 94:98 Epithelial Sites 83:98 Epithelial Sites 83:98 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 3 1 theme skin 713:716 arg1 glycans 724:730 the more simple skin mucin glycans 697:730 the more simple skin mucin glycans 697:730 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 5 2 contain have 875:878 arg1 acids 862:866 Sialic acids 855:866 Sialic acids 855:866 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 5 2 contain have 875:878 arg2 abundance 883:891 an abundance 880:891 an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans 880:968 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 7 3 theme interaction 1423:1433 arg1 determinants 1388:1399 important determinants 1378:1399 important determinants of the A. salmonicida interaction with its host at the mucosal surface 1378:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 3 theme interaction 1423:1433 arg1 GlcNAc 1350:1355 GlcNAc 1350:1355 GlcNAc 1350:1355 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 3 theme interaction 1423:1433 arg1 acids 1368:1372 sialic acids 1361:1372 sialic acids 1361:1372 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 6 4 dep mucins 1288:1293 arg1 response 1276:1283 response 1276:1283 response 1276:1283 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 3 5 theme glycan 525:530 arg1 residues 532:539 glycan residues 525:539 glycan residues 525:539 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 8 6 theme glycan 1549:1554 arg1 repertoire 1556:1565 the glycan repertoire 1545:1565 the glycan repertoire 1545:1565 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 8 6 theme glycan 1549:1554 arg1 factor 1576:1581 a factor 1574:1581 a factor to take into account during breeding and selection of strains for aquaculture 1574:1659 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 7 7 theme A. 1408:1409 arg1 interaction 1423:1433 the A. salmonicida interaction 1404:1433 the A. salmonicida interaction with its host at the mucosal surface 1404:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 4 8 theme present 763:769 arg1 residues 754:761 the glycan residues 743:761 the glycan residues present terminally on the salmon mucins 743:801 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 2 9 theme epithelial 303:312 arg1 surfaces 314:321 The epithelial surfaces 299:321 The epithelial surfaces that the pathogen colonizes 299:349 The epithelial surfaces that the pathogen colonizes are covered by a mucus layer predominantly comprised of secreted mucins. 28533470 7 10 theme mucosal 1456:1462 arg1 surface 1464:1470 the mucosal surface 1452:1470 the mucosal surface 1452:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 11 from determinants 1388:1399 arg1 conclusion 1338:1347 conclusion 1338:1347 conclusion 1338:1347 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 11 from determinants 1388:1399 arg1 surface 1464:1470 the mucosal surface 1452:1470 the mucosal surface 1452:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 1 12 theme Atlantic 270:277 arg1 aquaculture 286:296 Atlantic salmon aquaculture 270:296 Atlantic salmon aquaculture 270:296 Aeromonas salmonicida causes furunculosis in salmonids and is a threat to Atlantic salmon aquaculture. 28533470 7 13 from surface 1464:1470 arg1 determinants 1388:1399 important determinants 1378:1399 important determinants of the A. salmonicida interaction with its host at the mucosal surface 1378:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 13 from surface 1464:1470 arg1 acids 1368:1372 sialic acids 1361:1372 sialic acids 1361:1372 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 13 from surface 1464:1470 arg1 GlcNAc 1350:1355 GlcNAc 1350:1355 GlcNAc 1350:1355 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 13 from surface 1464:1470 arg1 interaction 1423:1433 the A. salmonicida interaction 1404:1433 the A. salmonicida interaction with its host at the mucosal surface 1404:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 5 14 theme % 943:943 arg1 abundance 883:891 an abundance 880:891 an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans 880:968 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 2 15 theme secreted 407:414 arg1 mucins 416:421 secreted mucins 407:421 secreted mucins 407:421 The epithelial surfaces that the pathogen colonizes are covered by a mucus layer predominantly comprised of secreted mucins. 28533470 3 16 theme mucin 718:722 arg1 glycans 724:730 the more simple skin mucin glycans 697:730 the more simple skin mucin glycans 697:730 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 8 17 theme pathogen 1528:1535 arg1 growth 1537:1542 pathogen growth 1528:1542 pathogen growth 1528:1542 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 7 18 theme important 1378:1386 arg1 determinants 1388:1399 important determinants 1378:1399 important determinants of the A. salmonicida interaction with its host at the mucosal surface 1378:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 18 theme important 1378:1386 arg1 GlcNAc 1350:1355 GlcNAc 1350:1355 GlcNAc 1350:1355 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 18 theme important 1378:1386 arg1 acids 1368:1372 sialic acids 1361:1372 sialic acids 1361:1372 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 0 19 theme salmonicida 10:20 arg1 Growth 22:27 Aeromonas salmonicida Growth 0:27 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins 0:65 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 6 20 from binding 1299:1305 arg1 mucins 1288:1293 mucins 1288:1293 mucins 1288:1293 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 6 21 theme growth-promoting 1192:1207 arg1 purposes 1209:1216 growth-promoting purposes 1192:1216 growth-promoting purposes 1192:1216 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 3 22 theme mass 433:436 arg1 spectrometry 438:449 mass spectrometry 433:449 mass spectrometry 433:449 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 0 23 theme Aeromonas 0:8 arg1 salmonicida 10:20 Aeromonas salmonicida 0:20 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins 0:65 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 1 24 theme salmon 279:284 arg1 aquaculture 286:296 Atlantic salmon aquaculture 270:296 Atlantic salmon aquaculture 270:296 Aeromonas salmonicida causes furunculosis in salmonids and is a threat to Atlantic salmon aquaculture. 28533470 7 25 theme sialic 1361:1366 arg1 determinants 1388:1399 important determinants 1378:1399 important determinants of the A. salmonicida interaction with its host at the mucosal surface 1378:1470 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 25 theme sialic 1361:1366 arg1 GlcNAc 1350:1355 GlcNAc 1350:1355 GlcNAc 1350:1355 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 7 25 theme sialic 1361:1366 arg1 acids 1368:1372 sialic acids 1361:1372 sialic acids 1361:1372 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 4 26 attach present 763:769 arg1 mucins 796:801 the salmon mucins 785:801 the salmon mucins 785:801 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 4 26 attach present 763:769 arg2 residues 754:761 the glycan residues 743:761 the glycan residues present terminally on the salmon mucins 743:801 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 0 27 theme Sialylated 116:125 arg1 O-Glycans 161:169 Sialylated and N-Acetylhexosamine-Containing O-Glycans 116:169 Sialylated and N-Acetylhexosamine-Containing O-Glycans 116:169 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 6 28 theme Ca2+ 1098:1101 arg1 concentration 1103:1115 A Ca2+ concentration 1096:1115 A Ca2+ concentration above 0.1 mM 1096:1128 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 6 28 theme Ca2+ 1098:1101 arg1 able 1165:1168 able 1165:1168 able 1165:1168 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 3 29 theme bacterial 569:577 arg1 growth 579:584 bacterial growth 569:584 bacterial growth 569:584 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 1 30 theme Aeromonas 196:204 arg1 salmonicida 206:216 Aeromonas salmonicida 196:216 Aeromonas salmonicida 196:216 Aeromonas salmonicida causes furunculosis in salmonids and is a threat to Atlantic salmon aquaculture. 28533470 5 31 theme growth-enhancing 1019:1034 arg1 purposes 1036:1043 growth-enhancing purposes 1019:1043 growth-enhancing purposes 1019:1043 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 0 32 dep Mucins 60:65 arg1 Response 32:39 Response 32:39 Response 32:39 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 5 33 used used 992:995 arg2 acids 862:866 Sialic acids 855:866 Sialic acids 855:866 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 8 34 theme strains 1637:1643 arg1 selection 1624:1632 selection 1624:1632 selection 1624:1632 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 8 34 theme strains 1637:1643 arg1 breeding 1611:1618 breeding 1611:1618 breeding 1611:1618 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 0 35 theme N-Acetylhexosamine-Containing 131:159 arg1 O-Glycans 161:169 Sialylated and N-Acetylhexosamine-Containing O-Glycans 116:169 Sialylated and N-Acetylhexosamine-Containing O-Glycans 116:169 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 3 36 theme A. 666:667 arg1 growth 681:686 A. salmonicida growth 666:686 A. salmonicida growth 666:686 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 3 37 theme mucin 463:467 arg1 structures 476:485 mucin glycan structures 463:485 mucin glycan structures 463:485 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 5 38 theme Sialic 855:860 arg1 acids 862:866 Sialic acids 855:866 Sialic acids 855:866 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 8 39 theme glycan 1502:1507 arg1 repertoire 1509:1518 the mucin glycan repertoire 1492:1518 the mucin glycan repertoire 1492:1518 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 3 40 theme mucin 644:648 arg1 glycans 650:656 the complex Atlantic salmon intestinal mucin glycans 605:656 the complex Atlantic salmon intestinal mucin glycans 605:656 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 4 41 theme only 804:807 arg1 N-acetylglucosamine 809:827 only N-acetylglucosamine 804:827 only N-acetylglucosamine (GlcNAc) 804:836 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 4 41 theme only 804:807 arg1 GlcNAc 830:835 GlcNAc 830:835 GlcNAc 830:835 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 3 42 theme glycan 469:474 arg1 structures 476:485 mucin glycan structures 463:485 mucin glycan structures 463:485 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 0 43 theme Salmon 53:58 arg1 Mucins 60:65 Atlantic Salmon Mucins 44:65 Atlantic Salmon Mucins 44:65 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 8 44 theme mucin 1496:1500 arg1 repertoire 1509:1518 the mucin glycan repertoire 1492:1518 the mucin glycan repertoire 1492:1518 Furthermore, since the mucin glycan repertoire affects pathogen growth, the glycan repertoire may be a factor to take into account during breeding and selection of strains for aquaculture. 28533470 6 45 from growth 1266:1271 arg1 mucins 1288:1293 mucins 1288:1293 mucins 1288:1293 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 1 46 from furunculosis 225:236 arg1 salmonids 241:249 salmonids 241:249 salmonids 241:249 Aeromonas salmonicida causes furunculosis in salmonids and is a threat to Atlantic salmon aquaculture. 28533470 3 47 theme enzymatic 504:512 arg1 removal 514:520 enzymatic removal 504:520 enzymatic removal of glycan residues 504:539 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 0 48 theme Atlantic 44:51 arg1 Mucins 60:65 Atlantic Salmon Mucins 44:65 Atlantic Salmon Mucins 44:65 Aeromonas salmonicida Growth in Response to Atlantic Salmon Mucins Differs between Epithelial Sites, Is Governed by Sialylated and N-Acetylhexosamine-Containing O-Glycans, and Is Affected by Ca2. 28533470 6 49 theme A. 1251:1252 arg1 growth 1266:1271 A. salmonicida growth 1251:1271 A. salmonicida growth in response to mucins 1251:1293 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 5 50 theme % 898:898 arg1 abundance 883:891 an abundance 880:891 an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans 880:968 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 5 51 from skin 928:931 arg1 glycans 915:921 terminal glycans 906:921 terminal glycans from skin 906:931 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 6 52 theme bacterium 1314:1322 arg1 binding 1299:1305 binding 1299:1305 binding of the bacterium to mucins 1299:1332 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 6 52 theme bacterium 1314:1322 arg1 growth 1266:1271 A. salmonicida growth 1251:1271 A. salmonicida growth in response to mucins 1251:1293 A Ca2+ concentration above 0.1 mM is needed for A. salmonicida to be able to utilize mucins for growth-promoting purposes, and 10 mM further enhances both A. salmonicida growth in response to mucins and binding of the bacterium to mucins. 28533470 4 53 theme salmon 789:794 arg1 mucins 796:801 the salmon mucins 785:801 the salmon mucins 785:801 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 3 54 theme salmon 626:631 arg1 glycans 650:656 the complex Atlantic salmon intestinal mucin glycans 605:656 the complex Atlantic salmon intestinal mucin glycans 605:656 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 4 55 theme glycan 747:752 arg1 residues 754:761 the glycan residues 743:761 the glycan residues present terminally on the salmon mucins 743:801 Of the glycan residues present terminally on the salmon mucins, only N-acetylglucosamine (GlcNAc) enhances growth. 28533470 3 56 theme intestinal 633:642 arg1 glycans 650:656 the complex Atlantic salmon intestinal mucin glycans 605:656 the complex Atlantic salmon intestinal mucin glycans 605:656 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 5 57 theme terminal 906:913 arg1 glycans 915:921 terminal glycans 906:921 terminal glycans from skin 906:931 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 3 58 theme growth 579:584 arg1 measurements 553:564 measurements 553:564 measurements of bacterial growth 553:584 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 7 59 with interaction 1423:1433 arg1 host 1444:1447 its host 1440:1447 its host 1440:1447 In conclusion, GlcNAc and sialic acids are important determinants of the A. salmonicida interaction with its host at the mucosal surface. 28533470 3 60 theme complex 609:615 arg1 salmon 626:631 complex Atlantic salmon 609:631 the complex Atlantic salmon intestinal mucin glycans 605:656 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 3 61 theme residues 532:539 arg1 removal 514:520 enzymatic removal 504:520 enzymatic removal of glycan residues 504:539 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 5 62 theme intestinal 951:960 arg1 glycans 962:968 intestinal glycans 951:968 intestinal glycans 951:968 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 3 63 theme simple 706:711 arg1 glycans 724:730 the more simple skin mucin glycans 697:730 the more simple skin mucin glycans 697:730 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 3 64 theme Atlantic 617:624 arg1 salmon 626:631 complex Atlantic salmon 609:631 the complex Atlantic salmon intestinal mucin glycans 605:656 By using mass spectrometry to identify mucin glycan structures with and without enzymatic removal of glycan residues, coupled to measurements of bacterial growth, we show here that the complex Atlantic salmon intestinal mucin glycans enhance A. salmonicida growth, whereas the more simple skin mucin glycans do not. 28533470 5 65 theme internal 1062:1069 arg1 GlcNAc 1071:1076 internal GlcNAc 1062:1076 internal GlcNAc 1062:1076 Sialic acids, which have an abundance of 75% among terminal glycans from skin and of <50% among intestinal glycans, cannot be removed or used by A. salmonicida for growth-enhancing purposes, and they shield internal GlcNAc from utilization. 28533470 2 66 theme mucus 368:372 arg1 layer 374:378 a mucus layer 366:378 a mucus layer predominantly comprised of secreted mucins 366:421 The epithelial surfaces that the pathogen colonizes are covered by a mucus layer predominantly comprised of secreted mucins. 26501333 4 0 theme sialic 771:776 arg1 acid 778:781 sialic acid 771:781 sialic acid containing glycans 771:800 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 0 1 theme Mass 99:102 arg1 Spectrometry 104:115 MALDI-Imaging Mass Spectrometry 85:115 MALDI-Imaging Mass Spectrometry 85:115 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 5 2 gly fucosylated 932:942 arg1 N-glycans 944:952 singly fucosylated N-glycans 925:952 singly fucosylated N-glycans detected in tumor tissue microarray cores 925:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 5 3 theme microarray 979:988 arg1 cores 990:994 tumor tissue microarray cores 966:994 tumor tissue microarray cores 966:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 1 4 theme two-dimensional 185:199 arg1 distribution 201:212 the two-dimensional distribution 181:212 the two-dimensional distribution of N-glycans in tissues 181:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 2 5 theme Assisted 290:297 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 5 theme Assisted 290:297 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 1 6 dep approach 150:157 arg1 map 177:179 map 177:179 to simultaneously map the two-dimensional distribution of N-glycans in tissues 159:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 6 7 theme technique 1236:1244 arg1 limitations 1217:1227 limitations 1217:1227 limitations 1217:1227 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 6 7 theme technique 1236:1244 arg1 advantages 1202:1211 advantages 1202:1211 advantages 1202:1211 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 8 dep location 390:397 arg1 the 386:388 the 386:388 the 386:388 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 9 theme Imaging 327:333 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 9 theme Imaging 327:333 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 5 10 theme fucosylated 932:942 arg1 N-glycans 944:952 singly fucosylated N-glycans 925:952 singly fucosylated N-glycans detected in tumor tissue microarray cores 925:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 3 11 theme hepatocellular 555:568 arg1 carcinoma 570:578 human hepatocellular carcinoma 549:578 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 3 12 theme Multiple 525:532 arg1 tissues 580:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 2 13 theme Ionization 316:325 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 13 theme Ionization 316:325 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 14 theme N-linked 428:435 arg1 species 444:450 multiple N-linked glycan species 419:450 multiple N-linked glycan species released by peptide N-glycosidase F 419:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 15 theme species 444:450 arg1 distribution 403:414 distribution 403:414 distribution 403:414 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 15 theme species 444:450 arg1 location 390:397 location 390:397 location 390:397 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 16 theme formalin-fixed 501:514 arg1 tissues 516:522 frozen or formalin-fixed tissues 491:522 frozen or formalin-fixed tissues 491:522 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 17 theme Desorption 305:314 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 17 theme Desorption 305:314 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 5 18 theme fucose 1059:1064 arg1 lectin 1074:1079 a core fucose binding lectin 1052:1079 a core fucose binding lectin 1052:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 2 19 theme glycan 437:442 arg1 species 444:450 multiple N-linked glycan species 419:450 multiple N-linked glycan species released by peptide N-glycosidase F 419:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 0 20 theme N-Glycan 16:23 arg1 Mapping 38:44 Two-Dimensional N-Glycan Distribution Mapping 0:44 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 0:116 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 4 21 contain containing 783:792 arg2 glycans 794:800 glycans 794:800 glycans 794:800 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 4 21 contain containing 783:792 arg1 acid 778:781 sialic acid 771:781 sialic acid containing glycans 771:800 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 6 22 theme tissue 1163:1168 arg1 block 1170:1174 any formalin-fixed tissue block 1144:1174 any formalin-fixed tissue block 1144:1174 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 23 from location 390:397 arg1 tissues 516:522 frozen or formalin-fixed tissues 491:522 frozen or formalin-fixed tissues 491:522 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 5 24 theme histochemistry 1017:1030 arg1 pattern 1041:1047 the histochemistry staining pattern 1013:1047 the histochemistry staining pattern of a core fucose binding lectin 1013:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 6 25 dep advantages 1202:1211 arg1 the 1198:1200 the 1198:1200 the 1198:1200 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 1 26 theme N-glycans 217:225 arg1 distribution 201:212 the two-dimensional distribution 181:212 the two-dimensional distribution of N-glycans in tissues 181:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 0 27 theme Two-Dimensional 0:14 arg1 Mapping 38:44 Two-Dimensional N-Glycan Distribution Mapping 0:44 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 0:116 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 6 28 with context 1249:1255 arg1 methods 1277:1283 other glycomic methods 1262:1283 other glycomic methods 1262:1283 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 5 29 theme staining 1032:1039 arg1 pattern 1041:1047 the histochemistry staining pattern 1013:1047 the histochemistry staining pattern of a core fucose binding lectin 1013:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 6 30 contain has 1109:1111 arg1 workflow 1100:1107 this MALDI-IMS workflow 1085:1107 this MALDI-IMS workflow 1085:1107 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 6 30 contain has 1109:1111 arg2 potential 1117:1125 the potential to be applied to any formalin-fixed tissue block or tissue microarray 1113:1195 the potential to be applied to any formalin-fixed tissue block or tissue microarray 1113:1195 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 31 theme multiple 419:426 arg1 species 444:450 multiple N-linked glycan species 419:450 multiple N-linked glycan species released by peptide N-glycosidase F 419:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 6 32 theme formalin-fixed 1148:1161 arg1 block 1170:1174 any formalin-fixed tissue block 1144:1174 any formalin-fixed tissue block 1144:1174 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 33 from distribution 403:414 arg1 tissues 516:522 frozen or formalin-fixed tissues 491:522 frozen or formalin-fixed tissues 491:522 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 1 34 theme new 120:122 arg1 approach 150:157 A new mass spectrometry imaging approach 118:157 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues 118:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 6 35 from limitations 1217:1227 arg1 context 1249:1255 context 1249:1255 context with other glycomic methods 1249:1283 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 36 theme N-glycosidase 472:484 arg1 F 486:486 peptide N-glycosidase F 464:486 peptide N-glycosidase F 464:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 0 37 theme Distribution 25:36 arg1 Mapping 38:44 Two-Dimensional N-Glycan Distribution Mapping 0:44 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 0:116 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 6 38 theme other 1262:1266 arg1 methods 1277:1283 other glycomic methods 1262:1283 other glycomic methods 1262:1283 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 2 39 theme frozen 491:496 arg1 tissues 516:522 frozen or formalin-fixed tissues 491:522 frozen or formalin-fixed tissues 491:522 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 40 link N-linked 428:435 arg1 species 444:450 multiple N-linked glycan species 419:450 multiple N-linked glycan species released by peptide N-glycosidase F 419:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 1 41 theme mass 124:127 arg1 approach 150:157 A new mass spectrometry imaging approach 118:157 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues 118:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 2 42 theme peptide 464:470 arg1 F 486:486 peptide N-glycosidase F 464:486 peptide N-glycosidase F 464:486 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 0 43 theme Hepatocellular 49:62 arg1 Tissues 74:80 Hepatocellular Carcinoma Tissues 49:80 Hepatocellular Carcinoma Tissues 49:80 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 3 44 theme carcinoma 570:578 arg1 tissues 580:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 3 45 theme N-glycans 653:661 arg1 panel 636:640 a panel 634:640 a panel of over 30 N-glycans detected 634:670 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 4 46 theme adjacent 733:740 arg1 slides 742:747 adjacent slides 733:747 adjacent slides 733:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 6 47 theme MALDI-IMS 1090:1098 arg1 workflow 1100:1107 this MALDI-IMS workflow 1085:1107 this MALDI-IMS workflow 1085:1107 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 4 48 theme N-glycans 709:717 arg1 reaction 687:694 An ethylation reaction 673:694 An ethylation reaction of extracted N-glycans released from adjacent slides 673:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 0 49 theme Tissues 74:80 arg1 Mapping 38:44 Two-Dimensional N-Glycan Distribution Mapping 0:44 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 0:116 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 5 50 located detected 954:961 arg2 N-glycans 944:952 singly fucosylated N-glycans 925:952 singly fucosylated N-glycans detected in tumor tissue microarray cores 925:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 5 50 located detected 954:961 arg1 cores 990:994 tumor tissue microarray cores 966:994 tumor tissue microarray cores 966:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 5 51 theme core 1054:1057 arg1 lectin 1074:1079 a core fucose binding lectin 1052:1079 a core fucose binding lectin 1052:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 6 52 theme tissue 1179:1184 arg1 microarray 1186:1195 tissue microarray 1179:1195 tissue microarray 1179:1195 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 1 53 theme spectrometry 129:140 arg1 approach 150:157 A new mass spectrometry imaging approach 118:157 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues 118:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 0 54 theme Carcinoma 64:72 arg1 Tissues 74:80 Hepatocellular Carcinoma Tissues 49:80 Hepatocellular Carcinoma Tissues 49:80 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 1 55 theme imaging 142:148 arg1 approach 150:157 A new mass spectrometry imaging approach 118:157 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues 118:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 6 56 theme glycomic 1268:1275 arg1 methods 1277:1283 other glycomic methods 1262:1283 other glycomic methods 1262:1283 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 5 57 theme lectin 1074:1079 arg1 pattern 1041:1047 the histochemistry staining pattern 1013:1047 the histochemistry staining pattern of a core fucose binding lectin 1013:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 2 58 theme Laser 299:303 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 58 theme Laser 299:303 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 6 59 from advantages 1202:1211 arg1 context 1249:1255 context 1249:1255 context with other glycomic methods 1249:1283 As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized. 26501333 3 60 theme formalin-fixed 534:547 arg1 tissues 580:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 5 61 theme binding 1066:1072 arg1 lectin 1074:1079 a core fucose binding lectin 1052:1079 a core fucose binding lectin 1052:1079 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 2 62 theme Mass 335:338 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 62 theme Mass 335:338 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 3 63 theme human 549:553 arg1 carcinoma 570:578 human hepatocellular carcinoma 549:578 Multiple formalin-fixed human hepatocellular carcinoma tissues 525:586 Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. 26501333 0 64 theme MALDI-Imaging 85:97 arg1 Spectrometry 104:115 MALDI-Imaging Mass Spectrometry 85:115 MALDI-Imaging Mass Spectrometry 85:115 Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry. 26501333 4 65 attach released 719:726 arg1 slides 742:747 adjacent slides 733:747 adjacent slides 733:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 4 65 attach released 719:726 arg2 N-glycans 709:717 extracted N-glycans 699:717 extracted N-glycans released from adjacent slides 699:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 2 66 theme Matrix 283:288 arg1 MALDI-IMS 354:362 MALDI-IMS 354:362 MALDI-IMS 354:362 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 2 66 theme Matrix 283:288 arg1 Spectrometry 340:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry 283:351 Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) 283:363 The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. 26501333 4 67 theme extracted 699:707 arg1 N-glycans 709:717 extracted N-glycans 699:717 extracted N-glycans released from adjacent slides 699:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 5 68 theme tumor 966:970 arg1 cores 990:994 tumor tissue microarray cores 966:994 tumor tissue microarray cores 966:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 5 69 theme N-glycans 944:952 arg1 distribution 909:920 the distribution 905:920 the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores 905:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 4 70 theme tissue 874:879 arg1 profiling 881:889 tissue profiling 874:889 tissue profiling 874:889 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 26501333 1 71 from distribution 201:212 arg1 tissues 230:236 tissues 230:236 tissues 230:236 A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. 26501333 5 72 theme tissue 972:977 arg1 cores 990:994 tumor tissue microarray cores 966:994 tumor tissue microarray cores 966:994 In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. 26501333 4 73 theme ethylation 676:685 arg1 reaction 687:694 An ethylation reaction 673:694 An ethylation reaction of extracted N-glycans released from adjacent slides 673:747 An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. 25359784 8 0 theme Asn448 1007:1012 arg1 chain 1019:1023 the Asn448 side chain 1003:1023 the Asn448 side chain 1003:1023 These results suggest that the Asn448 side chain is primarily important for the stability of IPU. 25359784 8 0 theme Asn448 1007:1012 arg1 important 1038:1046 important 1038:1046 important 1038:1046 These results suggest that the Asn448 side chain is primarily important for the stability of IPU. 25359784 7 1 theme wild-type 864:872 arg1 IPU 874:876 wild-type IPU 864:876 wild-type IPU 864:876 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 3 2 theme active 470:475 arg1 site 477:480 the active site 466:480 the active site 466:480 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 3 2 theme active 470:475 arg1 Asn448 483:488 Asn448 483:488 Asn448 483:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 5 3 theme Endo 652:655 arg1 H 657:657 Endo H 652:657 Endo H 652:657 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 3 4 theme site 477:480 arg1 vicinity 454:461 the vicinity 450:461 the vicinity of the active site, Asn448 450:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 2 5 gly N-glycosylated 261:274 arg1 protein 276:282 a highly N-glycosylated protein 252:282 a highly N-glycosylated protein 252:282 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 2 5 gly N-glycosylated 261:274 arg1 Isopullulanase 190:203 Isopullulanase 190:203 Isopullulanase (IPU) 190:209 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 10 6 theme atypical 1345:1352 arg1 structure 1354:1362 an atypical structure 1342:1362 an atypical structure that lacked this configuration 1342:1393 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 10 7 theme extended 1282:1289 arg1 configuration 1291:1303 an extended configuration 1279:1303 an extended configuration in IPU 1279:1310 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 5 8 theme N448A 668:672 arg1 variant 674:680 endoglycosidase H (Endo H)-treated N448A variant 633:680 endoglycosidase H (Endo H)-treated N448A variant 633:680 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 5 9 theme crystal 612:618 arg1 structure 620:628 The crystal structure 608:628 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant 608:680 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 2 10 from decrease 322:329 arg1 thermostability 334:348 thermostability 334:348 thermostability 334:348 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 2 11 theme N-glycosylated 261:274 arg1 protein 276:282 a highly N-glycosylated protein 252:282 a highly N-glycosylated protein 252:282 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 2 11 theme N-glycosylated 261:274 arg1 Isopullulanase 190:203 Isopullulanase 190:203 Isopullulanase (IPU) 190:209 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 9 12 theme N-glycan 1215:1222 arg1 function 1203:1210 the function 1199:1210 the function of N-glycan 1199:1222 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 10 13 from configuration 1291:1303 arg1 IPU 1308:1310 IPU 1308:1310 IPU 1308:1310 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 5 14 theme variant 674:680 arg1 structure 620:628 The crystal structure 608:628 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant 608:680 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 0 15 theme isopullulanase 86:99 arg1 stability 73:81 the stability 69:81 the stability of isopullulanase 69:99 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 3 16 from function 370:377 arg1 IPU 394:396 IPU 394:396 IPU 394:396 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 1 17 gly glycoproteins 175:187 arg1 glycoproteins 175:187 some glycoproteins 170:187 some glycoproteins 170:187 N-glycosylation has been shown to be important for the stability of some glycoproteins. 25359784 0 18 theme side 4:7 arg1 chain 9:13 The side chain 0:13 The side chain of a glycosylated asparagine residue 0:50 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 0 18 theme side 4:7 arg1 important 55:63 important 55:63 important 55:63 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 9 19 theme function 1203:1210 arg1 evaluation 1185:1194 the evaluation 1181:1194 the evaluation of the function of N-glycan 1181:1222 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 3 20 from vicinity 454:461 arg1 located 439:445 located 439:445 located 439:445 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 10 21 theme sites 1268:1272 arg1 structures 1238:1247 the structures 1234:1247 the structures of N-glycosylation sites 1234:1272 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 4 22 theme IPU 522:524 arg1 N448A 543:547 N448A 543:547 N448A 543:547 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 4 22 theme IPU 522:524 arg1 Y440A 536:540 Y440A 536:540 Y440A 536:540 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 4 22 theme IPU 522:524 arg1 variants 526:533 three IPU variants 516:533 three IPU variants 516:533 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 4 22 theme IPU 522:524 arg1 S450A 553:557 S450A 553:557 S450A 553:557 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 9 23 theme Asn 1132:1134 arg1 residue 1136:1142 only N-glycosylated Asn residue 1112:1142 only N-glycosylated Asn residue 1112:1142 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 6 24 theme IPU 713:715 arg1 molecules 717:725 four IPU molecules 708:725 four IPU molecules 708:725 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 6 24 theme IPU 713:715 arg1 D 744:744 D 744:744 D 744:744 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 6 24 theme IPU 713:715 arg1 C 738:738 C 738:738 C 738:738 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 6 24 theme IPU 713:715 arg1 B 735:735 B 735:735 B 735:735 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 6 24 theme IPU 713:715 arg1 Mol-A 728:732 Mol-A 728:732 Mol-A 728:732 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 2 25 theme IPU 289:291 arg1 deglycosylation 293:307 IPU deglycosylation 289:307 IPU deglycosylation 289:307 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 6 26 theme asymmetric 754:763 arg1 unit 765:768 the asymmetric unit 750:768 the asymmetric unit 750:768 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 10 27 gly N-glycosylation 1252:1266 arg2 sites 1268:1272 N-glycosylation sites 1252:1272 N-glycosylation sites 1252:1272 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 7 28 theme amino 810:814 arg1 residues 821:828 amino acid residues 435-455 810:836 amino acid residues 435-455 in Mol-C 810:845 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 29 theme acid 816:819 arg1 residues 821:828 amino acid residues 435-455 810:836 amino acid residues 435-455 in Mol-C 810:845 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 8 30 theme IPU 1069:1071 arg1 stability 1056:1064 the stability 1052:1064 the stability of IPU 1052:1071 These results suggest that the Asn448 side chain is primarily important for the stability of IPU. 25359784 3 31 theme located 439:445 arg1 residue 431:437 an N-glycosylated residue 413:437 an N-glycosylated residue located in the vicinity of the active site, Asn448 413:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 1 32 gly N-glycosylation 102:116 arg1 glycoproteins 175:187 some glycoproteins 170:187 some glycoproteins 170:187 N-glycosylation has been shown to be important for the stability of some glycoproteins. 25359784 0 33 theme glycosylated 20:31 arg1 residue 44:50 a glycosylated asparagine residue 18:50 a glycosylated asparagine residue 18:50 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 4 34 theme variants 526:533 arg1 thermostabilities 495:511 The thermostabilities 491:511 The thermostabilities of three IPU variants, Y440A, N448A and S450A, 491:558 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 4 34 theme variants 526:533 arg1 lower 575:579 lower 575:579 lower 575:579 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 4 35 theme wild-type 590:598 arg1 enzyme 600:605 the wild-type enzyme 586:605 the wild-type enzyme 586:605 The thermostabilities of three IPU variants, Y440A, N448A and S450A, were 0.5-8.4°C lower than the wild-type enzyme. 25359784 3 36 theme N-glycosylated 416:429 arg1 residue 431:437 an N-glycosylated residue 413:437 an N-glycosylated residue located in the vicinity of the active site, Asn448 413:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 7 37 from other 969:973 arg1 conformations 891:903 the conformations 887:903 the conformations of this loop in Mol-A, Mol-B and Mol-D 887:942 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 37 from other 969:973 arg1 different 949:957 different 949:957 different 949:957 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 10 38 theme Asn448 1326:1331 arg1 site 1333:1336 the Asn448 site 1322:1336 the Asn448 site 1322:1336 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 6 39 from molecules 717:725 arg1 unit 765:768 the asymmetric unit 750:768 the asymmetric unit 750:768 There are four IPU molecules, Mol-A, B, C and D, in the asymmetric unit. 25359784 3 40 theme N-glycan 382:389 arg1 function 370:377 the function 366:377 the function of N-glycan in IPU 366:396 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 5 41 theme -treated 659:666 arg1 variant 674:680 endoglycosidase H (Endo H)-treated N448A variant 633:680 endoglycosidase H (Endo H)-treated N448A variant 633:680 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 5 42 dep -treated 659:666 arg1 H 649:649 endoglycosidase H 633:649 endoglycosidase H 633:649 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 5 42 dep -treated 659:666 arg1 H 657:657 Endo H 652:657 Endo H 652:657 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 0 43 theme asparagine 33:42 arg1 residue 44:50 a glycosylated asparagine residue 18:50 a glycosylated asparagine residue 18:50 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 7 44 from conformations 891:903 arg1 Mol-B 928:932 Mol-B 928:932 Mol-B 928:932 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 44 from conformations 891:903 arg1 Mol-A 921:925 Mol-A 921:925 Mol-A 921:925 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 44 from conformations 891:903 arg1 Mol-D 938:942 Mol-D 938:942 Mol-D 938:942 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 3 45 gly N-glycosylated 416:429 arg1 residue 431:437 an N-glycosylated residue 413:437 an N-glycosylated residue located in the vicinity of the active site, Asn448 413:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 3 46 from located 439:445 arg1 vicinity 454:461 the vicinity 450:461 the vicinity of the active site, Asn448 450:488 To investigate the function of N-glycan in IPU, we focused on an N-glycosylated residue located in the vicinity of the active site, Asn448. 25359784 10 47 theme N-glycosylation 1252:1266 arg1 sites 1268:1272 N-glycosylation sites 1252:1272 N-glycosylation sites 1252:1272 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 5 48 theme endoglycosidase 633:647 arg1 H 649:649 endoglycosidase H 633:649 endoglycosidase H 633:649 The crystal structure of endoglycosidase H (Endo H)-treated N448A variant was determined. 25359784 7 49 from residues 821:828 arg1 Mol-C 841:845 Mol-C 841:845 Mol-C 841:845 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 50 theme loop 913:916 arg1 conformations 891:903 the conformations 887:903 the conformations of this loop in Mol-A, Mol-B and Mol-D 887:942 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 50 theme loop 913:916 arg1 different 949:957 different 949:957 different 949:957 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 9 51 theme N-glycosylated 1117:1130 arg1 residue 1136:1142 only N-glycosylated Asn residue 1112:1142 only N-glycosylated Asn residue 1112:1142 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 0 52 gly glycosylated 20:31 arg1 residue 44:50 a glycosylated asparagine residue 18:50 a glycosylated asparagine residue 18:50 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 10 53 contain had 1338:1340 arg2 structure 1354:1362 an atypical structure 1342:1362 an atypical structure that lacked this configuration 1342:1393 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 10 53 contain had 1338:1340 arg1 site 1333:1336 the Asn448 site 1322:1336 the Asn448 site 1322:1336 Usually, the structures of N-glycosylation sites form an extended configuration in IPU; however, the Asn448 site had an atypical structure that lacked this configuration. 25359784 9 54 gly N-glycosylated 1117:1130 arg1 residue 1136:1142 only N-glycosylated Asn residue 1112:1142 only N-glycosylated Asn residue 1112:1142 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 1 55 theme glycoproteins 175:187 arg1 stability 157:165 the stability 153:165 the stability of some glycoproteins 153:187 N-glycosylation has been shown to be important for the stability of some glycoproteins. 25359784 7 56 theme loop 793:796 arg1 conformation 775:786 The conformation 771:786 The conformation of a loop composed of amino acid residues 435-455 in Mol-C 771:845 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 7 56 theme loop 793:796 arg1 identical 851:859 identical 851:859 identical 851:859 The conformation of a loop composed of amino acid residues 435-455 in Mol-C was identical to wild-type IPU, whereas the conformations of this loop in Mol-A, Mol-B and Mol-D were different from each other. 25359784 0 57 theme residue 44:50 arg1 chain 9:13 The side chain 0:13 The side chain of a glycosylated asparagine residue 0:50 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 0 57 theme residue 44:50 arg1 important 55:63 important 55:63 important 55:63 The side chain of a glycosylated asparagine residue is important for the stability of isopullulanase. 25359784 9 58 theme residue 1136:1142 arg1 mutation 1100:1107 mutation 1100:1107 mutation of only N-glycosylated Asn residue 1100:1142 Our results indicate that mutation of only N-glycosylated Asn residue may lead to incorrect conclusion for the evaluation of the function of N-glycan. 25359784 2 59 theme polysaccharide-hydrolyzing 214:239 arg1 enzyme 241:246 a polysaccharide-hydrolyzing enzyme 212:246 a polysaccharide-hydrolyzing enzyme 212:246 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 2 59 theme polysaccharide-hydrolyzing 214:239 arg1 Isopullulanase 190:203 Isopullulanase 190:203 Isopullulanase (IPU) 190:209 Isopullulanase (IPU), a polysaccharide-hydrolyzing enzyme, is a highly N-glycosylated protein, and IPU deglycosylation results in a decrease in thermostability. 25359784 8 60 theme side 1014:1017 arg1 chain 1019:1023 the Asn448 side chain 1003:1023 the Asn448 side chain 1003:1023 These results suggest that the Asn448 side chain is primarily important for the stability of IPU. 25359784 8 60 theme side 1014:1017 arg1 important 1038:1046 important 1038:1046 important 1038:1046 These results suggest that the Asn448 side chain is primarily important for the stability of IPU. 25673493 7 0 theme capillary 1270:1278 arg1 CE 1297:1298 CE 1297:1298 CE 1297:1298 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 7 0 theme capillary 1270:1278 arg1 electrophoresis 1280:1294 capillary electrophoresis 1270:1294 capillary electrophoresis (CE)-based profiling 1270:1315 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 4 1 theme structural 612:621 arg1 diversity 623:631 The structural diversity 608:631 The structural diversity of these attached glycans 608:657 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 3 2 theme receptor-binding 531:546 arg1 activity 548:555 receptor-binding activity 531:555 receptor-binding activity 531:555 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 5 3 theme recent 921:926 arg1 target 928:933 a recent target 919:933 a recent target in efficacy and safety issues 919:963 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 2 4 gly glycoproteins 264:276 arg1 half 225:228 Over half 220:228 Over half of these new generation drugs 220:258 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 4 gly glycoproteins 264:276 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 4 gly glycoproteins 264:276 arg1 antibodies 298:307 monoclonal antibodies 287:307 monoclonal antibodies 287:307 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 4 gly glycoproteins 264:276 arg1 glycoproteins 264:276 glycoproteins 264:276 glycoproteins 264:276 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 5 theme other 312:316 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 6 6 theme clone 1127:1131 arg1 selection 1133:1141 clone selection 1127:1141 clone selection 1127:1141 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 5 7 theme modification 895:906 arg1 effects 864:870 the effects 860:870 the effects of their glycosylation modification 860:906 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 8 8 theme N-linked 1480:1487 arg1 glycans 1489:1495 the N-linked glycans 1476:1495 the N-linked glycans 1476:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 4 9 theme altered 680:686 arg1 composition 703:713 altered monosaccharide composition 680:713 altered monosaccharide composition 680:713 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 5 10 from target 928:933 arg1 issues 958:963 efficacy and safety issues 938:963 efficacy and safety issues 938:963 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 8 11 theme liberated 1514:1522 arg1 carbohydrates 1524:1536 the liberated carbohydrates 1510:1536 the liberated carbohydrates 1510:1536 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 5 12 theme biosimilar 810:819 arg1 products 821:828 more and more biosimilar products 796:828 more and more biosimilar products 796:828 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 2 13 theme generation 243:252 arg1 drugs 254:258 these new generation drugs 233:258 these new generation drugs 233:258 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 5 14 theme more 796:799 arg1 products 821:828 more and more biosimilar products 796:828 more and more biosimilar products 796:828 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 4 15 theme building 763:770 arg1 blocks 772:777 the monosaccharide building blocks 744:777 the monosaccharide building blocks 744:777 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 7 16 theme preparation 1243:1253 arg1 techniques 1255:1264 the recently introduced and commonly used sample preparation techniques 1194:1264 the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans 1194:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 8 17 theme mixture 1548:1554 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 18 theme protein 1390:1396 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 7 19 theme used 1231:1234 arg1 techniques 1255:1264 the recently introduced and commonly used sample preparation techniques 1194:1264 the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans 1194:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 6 20 theme manufacturing 1093:1105 arg1 process 1107:1113 the manufacturing process 1089:1113 the manufacturing process 1089:1113 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 8 21 theme affinity 1400:1407 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 0 22 theme Sample 0:5 arg1 preparation 7:17 Sample preparation 0:17 Sample preparation 0:17 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 8 23 link N-linked 1480:1487 arg1 glycans 1489:1495 the N-linked glycans 1476:1495 the N-linked glycans 1476:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 24 theme excess 1594:1599 arg1 reagent 1610:1616 the excess labeling reagent 1590:1616 the excess labeling reagent 1590:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 25 theme antibodies 1436:1445 arg1 release 1465:1471 enzymatic release 1455:1471 enzymatic release of the N-linked glycans 1455:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 25 theme antibodies 1436:1445 arg1 labeling 1498:1505 labeling 1498:1505 labeling of the liberated carbohydrates 1498:1536 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 25 theme antibodies 1436:1445 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 25 theme antibodies 1436:1445 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 25 theme antibodies 1436:1445 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 3 26 theme thermal 482:488 arg1 stability 490:498 thermal stability 482:498 thermal stability 482:498 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 1 27 theme considerable 117:128 arg1 number 130:135 a considerable number 115:135 a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade 115:217 There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. 25673493 7 28 theme N-glycans 1347:1355 arg1 profiling 1307:1315 capillary electrophoresis (CE)-based profiling 1270:1315 capillary electrophoresis (CE)-based profiling 1270:1315 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 7 28 theme N-glycans 1347:1355 arg1 elucidation 1332:1342 structural elucidation 1321:1342 structural elucidation of N-glycans 1321:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 6 29 dep selection 1133:1141 arg1 i.e. 1116:1119 i.e. 1116:1119 i.e. 1116:1119 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 3 30 theme linked 400:405 arg1 moieties 420:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 0 31 theme antibodies 74:83 arg1 analysis 39:46 N-glycosylation analysis 23:46 N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis 23:102 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 3 32 theme crucial 509:515 arg1 features 517:524 crucial features 509:524 not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity 436:555 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 0 33 theme therapeutic 51:61 arg1 antibodies 74:83 therapeutic monoclonal antibodies 51:83 therapeutic monoclonal antibodies 51:83 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 0 34 gly N-glycosylation 23:37 arg1 antibodies 74:83 therapeutic monoclonal antibodies 51:83 therapeutic monoclonal antibodies 51:83 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 2 35 gly glycoproteins 330:342 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 7 36 theme -based 1300:1305 arg1 profiling 1307:1315 capillary electrophoresis (CE)-based profiling 1270:1315 capillary electrophoresis (CE)-based profiling 1270:1315 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 2 37 theme mammalian 374:382 arg1 lines 389:393 mammalian cell lines 374:393 mammalian cell lines 374:393 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 8 38 theme labeled 1734:1740 arg1 N-glycans 1755:1763 the labeled and purified N-glycans 1730:1763 N-glycans 1755:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 6 39 theme consistent 976:985 arg1 quality 987:993 consistent quality 976:993 consistent quality of these products 976:1011 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 4 40 theme glycans 651:657 arg1 diversity 623:631 The structural diversity 608:631 The structural diversity of these attached glycans 608:657 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 8 41 theme purified 1746:1753 arg1 N-glycans 1755:1763 the labeled and purified N-glycans 1730:1763 N-glycans 1755:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 7 42 theme electrophoresis 1280:1294 arg1 profiling 1307:1315 capillary electrophoresis (CE)-based profiling 1270:1315 capillary electrophoresis (CE)-based profiling 1270:1315 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 2 43 theme monoclonal 287:296 arg1 antibodies 298:307 monoclonal antibodies 287:307 monoclonal antibodies 287:307 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 8 44 theme -based 1710:1715 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 1 45 theme clinical 187:194 arg1 use 196:198 clinical use 187:198 clinical use in the past decade 187:217 There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. 25673493 4 46 theme monosaccharide 688:701 arg1 composition 703:713 altered monosaccharide composition 680:713 altered monosaccharide composition 680:713 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 2 47 theme recombinant 318:328 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 5 48 theme glycosylation 881:893 arg1 modification 895:906 their glycosylation modification 875:906 their glycosylation modification 875:906 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 6 49 theme glycosylation 1014:1026 arg1 profiles 1028:1035 glycosylation profiles 1014:1035 glycosylation profiles 1014:1035 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 8 50 theme glycans 1489:1495 arg1 release 1465:1471 enzymatic release 1455:1471 enzymatic release of the N-linked glycans 1455:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 50 theme glycans 1489:1495 arg1 labeling 1498:1505 labeling 1498:1505 labeling of the liberated carbohydrates 1498:1536 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 50 theme glycans 1489:1495 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 50 theme glycans 1489:1495 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 50 theme glycans 1489:1495 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 51 theme presented 1362:1370 arg1 protocols 1372:1380 The presented protocols 1358:1380 The presented protocols 1358:1380 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 3 52 link linked 400:405 arg1 moieties 420:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 6 53 theme products 1004:1011 arg1 quality 987:993 consistent quality 976:993 consistent quality of these products 976:1011 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 1 54 theme biopharmaceuticals 140:157 arg1 number 130:135 a considerable number 115:135 a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade 115:217 There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. 25673493 8 55 theme carbohydrates 1524:1536 arg1 release 1465:1471 enzymatic release 1455:1471 enzymatic release of the N-linked glycans 1455:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 55 theme carbohydrates 1524:1536 arg1 labeling 1498:1505 labeling 1498:1505 labeling of the liberated carbohydrates 1498:1536 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 55 theme carbohydrates 1524:1536 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 55 theme carbohydrates 1524:1536 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 55 theme carbohydrates 1524:1536 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 2 56 theme drugs 254:258 arg1 half 225:228 Over half 220:228 Over half of these new generation drugs 220:258 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 56 theme drugs 254:258 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 56 theme drugs 254:258 arg1 antibodies 298:307 monoclonal antibodies 287:307 monoclonal antibodies 287:307 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 56 theme drugs 254:258 arg1 glycoproteins 264:276 glycoproteins 264:276 glycoproteins 264:276 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 1 57 theme past 207:210 arg1 decade 212:217 the past decade 203:217 the past decade 203:217 There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. 25673493 5 58 theme efficacy 938:945 arg1 issues 958:963 efficacy and safety issues 938:963 efficacy and safety issues 938:963 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 8 59 theme reaction 1539:1546 arg1 mixture 1548:1554 reaction mixture 1539:1554 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 7 60 theme sample 1236:1241 arg1 techniques 1255:1264 the recently introduced and commonly used sample preparation techniques 1194:1264 the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans 1194:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 1 61 from use 196:198 arg1 decade 212:217 the past decade 203:217 the past decade 203:217 There are a considerable number of biopharmaceuticals that have been approved for clinical use in the past decade. 25673493 5 62 theme safety 951:956 arg1 issues 958:963 efficacy and safety issues 938:963 efficacy and safety issues 938:963 In addition, as more and more biosimilar products hit the market, understanding the effects of their glycosylation modification has become a recent target in efficacy and safety issues. 25673493 8 63 theme purification 1556:1567 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 4 64 theme monosaccharide 748:761 arg1 blocks 772:777 the monosaccharide building blocks 744:777 the monosaccharide building blocks 744:777 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 7 65 theme introduced 1207:1216 arg1 techniques 1255:1264 the recently introduced and commonly used sample preparation techniques 1194:1264 the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans 1194:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 8 66 theme A 1398:1398 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 3 67 theme physicochemical 451:465 arg1 properties 467:476 physicochemical properties 451:476 physicochemical properties 451:476 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 8 68 theme labeling 1601:1608 arg1 reagent 1610:1616 the excess labeling reagent 1590:1616 the excess labeling reagent 1590:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 2 69 theme cell 384:387 arg1 lines 389:393 mammalian cell lines 374:393 mammalian cell lines 374:393 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 0 70 theme N-glycosylation 23:37 arg1 analysis 39:46 N-glycosylation analysis 23:46 N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis 23:102 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 2 71 theme new 239:241 arg1 drugs 254:258 these new generation drugs 233:258 these new generation drugs 233:258 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 6 72 theme process 1107:1113 arg1 steps 1080:1084 all steps 1076:1084 all steps 1076:1084 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 8 73 theme monoclonal 1425:1434 arg1 mAbs 1448:1451 mAbs 1448:1451 mAbs 1448:1451 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 73 theme monoclonal 1425:1434 arg1 antibodies 1436:1445 monoclonal antibodies 1425:1445 monoclonal antibodies (mAbs) 1425:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 7 74 theme structural 1321:1330 arg1 elucidation 1332:1342 structural elucidation 1321:1342 structural elucidation of N-glycans 1321:1355 In this paper, we describe some of the recently introduced and commonly used sample preparation techniques for capillary electrophoresis (CE)-based profiling and structural elucidation of N-glycans. 25673493 8 75 theme high-resolution 1623:1637 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 0 76 theme monoclonal 63:72 arg1 antibodies 74:83 therapeutic monoclonal antibodies 51:83 therapeutic monoclonal antibodies 51:83 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 3 77 theme carbohydrate 407:418 arg1 moieties 420:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties 396:427 The linked carbohydrate moieties affect not only their physicochemical properties and thermal stability but also crucial features like receptor-binding activity, circulating half-life, as well as immunogenicity. 25673493 8 78 theme rapid 1643:1647 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 2 79 theme Over 220:223 arg1 half 225:228 Over half 220:228 Over half of these new generation drugs 220:258 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 79 theme Over 220:223 arg1 glycoproteins 330:342 other recombinant glycoproteins 312:342 other recombinant glycoproteins 312:342 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 79 theme Over 220:223 arg1 antibodies 298:307 monoclonal antibodies 287:307 monoclonal antibodies 287:307 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 2 79 theme Over 220:223 arg1 glycoproteins 264:276 glycoproteins 264:276 glycoproteins 264:276 Over half of these new generation drugs are glycoproteins, such as monoclonal antibodies or other recombinant glycoproteins, which are mostly produced in mammalian cell lines. 25673493 4 80 theme attached 642:649 arg1 glycans 651:657 these attached glycans 636:657 these attached glycans 636:657 The structural diversity of these attached glycans can be manifested in altered monosaccharide composition and linkages/positions among the monosaccharide building blocks. 25673493 8 81 theme enzymatic 1455:1463 arg1 release 1465:1471 enzymatic release 1455:1471 enzymatic release of the N-linked glycans 1455:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 0 82 dep preparation 7:17 arg1 analysis 39:46 N-glycosylation analysis 23:46 N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis 23:102 Sample preparation for N-glycosylation analysis of therapeutic monoclonal antibodies by electrophoresis. 25673493 6 83 theme lot 1146:1148 arg1 release 1150:1156 lot release 1146:1156 lot release 1146:1156 To ensure consistent quality of these products, glycosylation profiles have to be monitored and controlled in all steps of the manufacturing process, i.e., from clone selection to lot release. 25673493 8 84 theme N-glycans 1755:1763 arg1 release 1465:1471 enzymatic release 1455:1471 enzymatic release of the N-linked glycans 1455:1495 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 84 theme N-glycans 1755:1763 arg1 labeling 1498:1505 labeling 1498:1505 labeling of the liberated carbohydrates 1498:1536 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 84 theme N-glycans 1755:1763 arg1 techniques 1569:1578 reaction mixture purification techniques 1539:1578 reaction mixture purification techniques to remove the excess labeling reagent 1539:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 84 theme N-glycans 1755:1763 arg1 partitioning 1409:1420 protein A affinity partitioning 1390:1420 protein A affinity partitioning of monoclonal antibodies (mAbs) 1390:1452 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 84 theme N-glycans 1755:1763 arg1 profiling 1717:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling 1623:1725 high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans 1623:1763 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 25673493 8 85 dep techniques 1569:1578 arg1 remove 1583:1588 remove 1583:1588 to remove the excess labeling reagent 1580:1616 The presented protocols include protein A affinity partitioning of monoclonal antibodies (mAbs), enzymatic release of the N-linked glycans, labeling of the liberated carbohydrates, reaction mixture purification techniques to remove the excess labeling reagent, and high-resolution and rapid capillary electrophoresis-laser-induced fluorescence (CE-LIF)-based profiling of the labeled and purified N-glycans. 28749929 6 0 theme site-specific 1484:1496 arg1 conjugation 1512:1522 site-specific antibody-drug conjugation 1484:1522 site-specific antibody-drug conjugation 1484:1522 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 1 theme antibody-related 1359:1374 arg1 research 1376:1383 antibody-related research 1359:1383 antibody-related research 1359:1383 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 4 2 theme IgG 700:702 arg1 GlcNAc 689:694 GlcNAc 689:694 GlcNAc 689:694 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 2 theme IgG 700:702 arg1 N-acetylglucosamine 668:686 the remaining N-acetylglucosamine 654:686 the remaining N-acetylglucosamine (GlcNAc) of IgG 654:702 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 5 3 theme produced 1070:1077 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 4 dep in 951:952 arg1 vitro 954:958 vitro 954:958 vitro 954:958 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 2 5 theme powerful 360:367 arg1 technology 344:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 5 theme powerful 360:367 arg1 tool 369:372 a powerful tool 358:372 a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling 358:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 6 6 theme therapeutic 1437:1447 arg1 antibodies 1449:1458 novel glycoengineered therapeutic antibodies 1415:1458 novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1415:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 2 7 theme fragment 386:393 arg1 remodeling 426:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 7 theme fragment 386:393 arg1 cystallizable 395:407 fragment cystallizable 386:407 fragment cystallizable 386:407 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 6 8 theme method 1329:1334 arg1 applications 1308:1319 Potential applications 1298:1319 Potential applications of this method 1298:1334 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 5 9 with compatibility 1100:1112 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 6 10 theme antibodies 1559:1568 arg1 conjugation 1512:1522 site-specific antibody-drug conjugation 1484:1522 site-specific antibody-drug conjugation 1484:1522 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 10 theme antibodies 1559:1568 arg1 modification 1543:1554 site-specific modification 1529:1554 site-specific modification 1529:1554 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 10 theme antibodies 1559:1568 arg1 efficacy 1474:1481 enhanced efficacy 1465:1481 enhanced efficacy 1465:1481 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 2 11 theme Fc 382:383 arg1 remodeling 426:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 11 theme Fc 382:383 arg1 cystallizable 395:407 fragment cystallizable 386:407 fragment cystallizable 386:407 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 6 12 theme glycoengineered 1421:1435 arg1 antibodies 1449:1458 novel glycoengineered therapeutic antibodies 1415:1458 novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1415:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 5 13 theme well-defined 1240:1251 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 13 theme well-defined 1240:1251 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 2 14 theme in 318:319 arg1 technology 344:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 14 theme in 318:319 arg1 tool 369:372 a powerful tool 358:372 a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling 358:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 5 15 theme antibodies 1079:1088 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 15 theme antibodies 1079:1088 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 16 theme ADCs 1094:1097 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 7 17 theme IgG 1803:1805 arg1 N-glycosylation 1807:1821 the IgG N-glycosylation 1799:1821 the IgG N-glycosylation 1799:1821 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 5 18 theme glycoengineering 885:900 arg1 technologies 902:913 in vivo glycoengineering technologies 877:913 in vivo glycoengineering technologies 877:913 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 2 19 theme glycoengineering 327:342 arg1 technology 344:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 19 theme glycoengineering 327:342 arg1 tool 369:372 a powerful tool 358:372 a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling 358:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 5 20 theme homogeneous 1044:1054 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 20 theme homogeneous 1044:1054 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 21 with glycoforms 1056:1065 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 1 22 theme great 219:223 arg1 interest 225:232 great interest 219:232 great interest among researchers 219:250 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 1 23 theme glycosite-specific 151:168 arg1 conjugates 184:193 glycosite-specific antibody-drug conjugates 151:193 glycosite-specific antibody-drug conjugates 151:193 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 4 24 theme homogeneous 573:583 arg1 substrate 594:602 a homogeneous N-glycan substrate 571:602 a homogeneous N-glycan substrate 571:602 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 24 theme homogeneous 573:583 arg1 presynthesized 605:618 presynthesized 605:618 presynthesized 605:618 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 3 25 theme wild-type 538:546 arg1 endoglycosidase 548:562 a wild-type endoglycosidase 536:562 a wild-type endoglycosidase 536:562 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 5 26 theme in 877:878 arg1 technologies 902:913 in vivo glycoengineering technologies 877:913 in vivo glycoengineering technologies 877:913 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 27 theme antibody 1274:1281 arg1 modifications 1283:1295 antibody modifications 1274:1295 antibody modifications 1274:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 3 28 gly glycosylated 479:490 arg1 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 5 29 theme glycan 1151:1156 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 3 30 theme glycosylated 479:490 arg1 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 0 31 theme Chemoenzymatic 0:13 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates 0:105 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 3 32 theme native 456:461 arg1 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 7 33 theme modified 1751:1758 arg1 substrates 1769:1778 the natural or modified N-glycan substrates 1736:1778 the natural or modified N-glycan substrates 1736:1778 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 5 34 theme diverse 1119:1125 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 7 35 theme small-molecule 1852:1865 arg1 toxins 1867:1872 the small-molecule toxins 1848:1872 the small-molecule toxins 1848:1872 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 0 36 theme IgG 44:46 arg1 antibodies 48:57 glycoengineered IgG antibodies 28:57 glycoengineered IgG antibodies 28:57 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 5 37 theme glycosyltransferase-enabled 923:949 arg1 method 972:977 the glycosyltransferase-enabled in vitro engineering method 919:977 the glycosyltransferase-enabled in vitro engineering method 919:977 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 7 38 theme natural 1740:1746 arg1 substrates 1769:1778 the natural or modified N-glycan substrates 1736:1778 the natural or modified N-glycan substrates 1736:1778 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 5 39 theme native 1197:1202 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 0 40 theme antibody-drug 82:94 arg1 conjugates 96:105 glycosite-specific antibody-drug conjugates 63:105 glycosite-specific antibody-drug conjugates 63:105 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 6 41 theme immunoliposome 1631:1644 arg1 formation 1646:1654 immunoliposome formation 1631:1654 immunoliposome formation 1631:1654 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 41 theme immunoliposome 1631:1644 arg1 labeling 1586:1593 fluorescent labeling 1574:1593 fluorescent labeling 1574:1593 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 4 42 dep endoglycosidase 720:734 arg1 called 742:747 called 742:747 called endoglycosynthase 742:765 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 2 43 theme N-glycosylation 410:424 arg1 remodeling 426:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 43 theme N-glycosylation 410:424 arg1 cystallizable 395:407 fragment cystallizable 386:407 fragment cystallizable 386:407 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 0 44 theme glycosite-specific 63:80 arg1 conjugates 96:105 glycosite-specific antibody-drug conjugates 63:105 glycosite-specific antibody-drug conjugates 63:105 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 4 45 contain possesses 803:811 arg1 endoglycosidase 720:734 a mutant endoglycosidase 711:734 a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering 711:860 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 45 contain possesses 803:811 arg2 activity 832:839 transglycosylation activity 813:839 transglycosylation activity for glycoengineering 813:860 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 6 46 theme antibody-drug 1498:1510 arg1 conjugation 1512:1522 site-specific antibody-drug conjugation 1484:1522 site-specific antibody-drug conjugation 1484:1522 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 47 theme antigen-binding 1684:1698 arg1 affinity 1700:1707 antigen-binding affinity 1684:1707 antigen-binding affinity 1684:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 2 48 theme IgG 378:380 arg1 Fc 382:383 IgG Fc 378:383 IgG Fc (fragment cystallizable) N-glycosylation remodeling 378:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 6 49 theme enhanced 1465:1472 arg1 efficacy 1474:1481 enhanced efficacy 1465:1481 enhanced efficacy 1465:1481 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 50 theme broad 1344:1348 arg1 scope 1350:1354 a broad scope 1342:1354 a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1342:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 51 theme antibodies 1449:1458 arg1 development 1400:1410 the development 1396:1410 the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1396:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 5 52 theme quantitative 1024:1035 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 52 theme quantitative 1024:1035 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 4 53 attach attached 642:649 arg1 GlcNAc 689:694 GlcNAc 689:694 GlcNAc 689:694 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 53 attach attached 642:649 arg1 N-acetylglucosamine 668:686 the remaining N-acetylglucosamine 654:686 the remaining N-acetylglucosamine (GlcNAc) of IgG 654:702 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 53 attach attached 642:649 arg2 presynthesized 605:618 presynthesized 605:618 presynthesized 605:618 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 53 attach attached 642:649 arg2 substrate 594:602 a homogeneous N-glycan substrate 571:602 a homogeneous N-glycan substrate 571:602 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 5 54 theme current 984:990 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 54 theme current 984:990 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 1 55 theme therapeutic 269:279 arg1 potential 281:289 their therapeutic potential 263:289 their therapeutic potential 263:289 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 6 56 theme novel 1415:1419 arg1 antibodies 1449:1458 novel glycoengineered therapeutic antibodies 1415:1458 novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1415:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 2 57 dep in 318:319 arg1 vitro 321:325 vitro 321:325 vitro 321:325 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 6 58 theme site-specific 1529:1541 arg1 modification 1543:1554 site-specific modification 1529:1554 site-specific modification 1529:1554 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 5 59 theme conjugation 1253:1263 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 59 theme conjugation 1253:1263 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 2 60 theme Endoglycosidase-catalyzed 292:316 arg1 technology 344:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology 292:353 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 2 60 theme Endoglycosidase-catalyzed 292:316 arg1 tool 369:372 a powerful tool 358:372 a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling 358:435 Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. 28749929 1 61 theme antibody-drug 170:182 arg1 conjugates 184:193 glycosite-specific antibody-drug conjugates 151:193 glycosite-specific antibody-drug conjugates 151:193 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 5 62 dep in 877:878 arg1 vivo 880:883 vivo 880:883 vivo 880:883 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 4 63 theme transglycosylation 813:830 arg1 activity 832:839 transglycosylation activity 813:839 transglycosylation activity for glycoengineering 813:860 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 5 64 theme starting 1215:1222 arg1 material 1224:1231 the starting material 1211:1231 the starting material 1211:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 65 gly glycoforms 1056:1065 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 65 gly glycoforms 1056:1065 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 65 gly glycoforms 1056:1065 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 66 with site 1265:1268 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 0 67 theme conjugates 96:105 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates 0:105 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 6 68 theme research 1376:1383 arg1 scope 1350:1354 a broad scope 1342:1354 a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity 1342:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 4 69 theme hydrolytic 779:788 arg1 activity 790:797 hydrolytic activity 779:797 hydrolytic activity 779:797 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 6 70 theme protein 1608:1614 arg1 cross-linking 1616:1628 protein cross-linking 1608:1628 protein cross-linking 1608:1628 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 70 theme protein 1608:1614 arg1 labeling 1586:1593 fluorescent labeling 1574:1593 fluorescent labeling 1574:1593 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 3 71 gly deglycosylated 516:529 arg1 protocol 446:453 this protocol 441:453 this protocol 441:453 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 3 71 gly deglycosylated 516:529 arg2 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 3 71 gly deglycosylated 516:529 arg1 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 7 72 theme 3-4 1781:1783 arg1 d 1785:1785 3-4 d 1781:1785 3-4 d to engineer the IgG N-glycosylation 1781:1821 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 5 73 theme natural 1127:1133 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 4 74 theme N-glycan 585:592 arg1 substrate 594:602 a homogeneous N-glycan substrate 571:602 a homogeneous N-glycan substrate 571:602 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 74 theme N-glycan 585:592 arg1 presynthesized 605:618 presynthesized 605:618 presynthesized 605:618 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 5 75 theme non-natural 1139:1149 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 76 theme engineering 960:970 arg1 method 972:977 the glycosyltransferase-enabled in vitro engineering method 919:977 the glycosyltransferase-enabled in vitro engineering method 919:977 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 1 77 theme therapeutic 124:134 arg1 antibodies 136:145 Glycoengineered therapeutic antibodies 108:145 Glycoengineered therapeutic antibodies 108:145 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 1 77 theme therapeutic 124:134 arg1 gsADCs 196:201 gsADCs 196:201 gsADCs 196:201 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 3 78 theme IgG 492:494 arg1 N-glycans 496:504 native heterogeneously glycosylated IgG N-glycans 456:504 native heterogeneously glycosylated IgG N-glycans 456:504 In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. 28749929 4 79 theme remaining 658:666 arg1 GlcNAc 689:694 GlcNAc 689:694 GlcNAc 689:694 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 4 79 theme remaining 658:666 arg1 N-acetylglucosamine 668:686 the remaining N-acetylglucosamine 654:686 the remaining N-acetylglucosamine (GlcNAc) of IgG 654:702 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 0 80 theme glycoengineered 28:42 arg1 antibodies 48:57 glycoengineered IgG antibodies 28:57 glycoengineered IgG antibodies 28:57 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 0 81 theme antibodies 48:57 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates 0:105 Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates. 28749929 6 82 dep and 1657:1659 arg1 so 1661:1662 so 1661:1662 so 1661:1662 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 7 83 theme N-glycan 1760:1767 arg1 substrates 1769:1778 the natural or modified N-glycan substrates 1736:1778 the natural or modified N-glycan substrates 1736:1778 It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs. 28749929 4 84 dep called 742:747 arg1 also 737:740 also 737:740 also 737:740 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 6 85 theme fluorescent 1574:1584 arg1 PEGylation 1596:1605 PEGylation 1596:1605 PEGylation 1596:1605 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 85 theme fluorescent 1574:1584 arg1 cross-linking 1616:1628 protein cross-linking 1608:1628 protein cross-linking 1608:1628 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 85 theme fluorescent 1574:1584 arg1 labeling 1586:1593 fluorescent labeling 1574:1593 fluorescent labeling 1574:1593 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 85 theme fluorescent 1574:1584 arg1 formation 1646:1654 immunoliposome formation 1631:1654 immunoliposome formation 1631:1654 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 5 86 theme in 951:952 arg1 method 972:977 the glycosyltransferase-enabled in vitro engineering method 919:977 the glycosyltransferase-enabled in vitro engineering method 919:977 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 87 theme IgG 1204:1206 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 88 with yield 1037:1041 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 4 89 dep presynthesized 605:618 arg1 described 623:631 described 623:631 described here 623:636 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 6 90 theme Potential 1298:1306 arg1 applications 1308:1319 Potential applications 1298:1319 Potential applications of this method 1298:1334 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 1 91 theme Glycoengineered 108:122 arg1 antibodies 136:145 Glycoengineered therapeutic antibodies 108:145 Glycoengineered therapeutic antibodies 108:145 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 1 91 theme Glycoengineered 108:122 arg1 gsADCs 196:201 gsADCs 196:201 gsADCs 196:201 Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. 28749929 5 92 dep robust 1004:1009 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 92 dep robust 1004:1009 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 antibodies 1079:1088 produced antibodies 1070:1088 produced antibodies 1070:1088 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 exploitation 1181:1192 convenient exploitation 1170:1192 convenient exploitation of native IgG as the starting material 1170:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 approach 992:999 the current approach 980:999 the current approach 980:999 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 ADCs 1094:1097 ADCs 1094:1097 ADCs 1094:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 site 1265:1268 a well-defined conjugation site 1238:1268 a well-defined conjugation site for antibody modifications 1238:1295 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 robust 1004:1009 robust 1004:1009 robust 1004:1009 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 glycoforms 1056:1065 homogeneous glycoforms 1044:1065 homogeneous glycoforms of produced antibodies and ADCs 1044:1097 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 IgG 1204:1206 native IgG 1197:1206 native IgG as the starting material 1197:1231 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 compatibility 1100:1112 compatibility 1100:1112 compatibility with diverse natural and non-natural glycan structures 1100:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 5 93 theme convenient 1170:1179 arg1 yield 1037:1041 quantitative yield 1024:1041 quantitative yield 1024:1041 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 6 94 with antibodies 1449:1458 arg1 conjugation 1512:1522 site-specific antibody-drug conjugation 1484:1522 site-specific antibody-drug conjugation 1484:1522 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 94 with antibodies 1449:1458 arg1 efficacy 1474:1481 enhanced efficacy 1465:1481 enhanced efficacy 1465:1481 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 6 94 with antibodies 1449:1458 arg1 modification 1543:1554 site-specific modification 1529:1554 site-specific modification 1529:1554 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 28749929 4 95 theme mutant 713:718 arg1 endoglycosidase 720:734 a mutant endoglycosidase 711:734 a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering 711:860 Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. 28749929 5 96 with exploitation 1181:1192 arg1 structures 1158:1167 diverse natural and non-natural glycan structures 1119:1167 diverse natural and non-natural glycan structures 1119:1167 Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. 28749929 6 97 theme affinity 1700:1707 arg1 loss 1676:1679 loss 1676:1679 loss of antigen-binding affinity 1676:1707 Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. 26066491 8 0 theme sialic 1043:1048 arg1 Neu5Gc 1065:1070 Neu5Gc 1065:1070 Neu5Gc 1065:1070 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 0 theme sialic 1043:1048 arg1 Kdn 1077:1079 Kdn 1077:1079 Kdn 1077:1079 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 0 theme sialic 1043:1048 arg1 Neu5Ac 1057:1062 Neu5Ac 1057:1062 Neu5Ac 1057:1062 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 0 theme sialic 1043:1048 arg1 acids 1050:1054 sialic acids 1043:1054 sialic acids (Neu5Ac, Neu5Gc, and Kdn) 1043:1080 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 2 1 theme contact 324:330 arg1 skin 230:233 The skin 226:233 The skin 226:233 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 1 theme contact 324:330 arg1 surfaces 247:254 mucosal surfaces 239:254 mucosal surfaces 239:254 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 1 theme contact 324:330 arg1 point 315:319 the first point 305:319 the first point of contact between fish and pathogens 305:357 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 10 2 theme host-pathogen 1374:1386 arg1 interactions 1388:1399 host-pathogen interactions 1374:1399 host-pathogen interactions for further testing in biological assays and disease intervention therapies 1374:1475 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 6 3 theme pyloric 791:797 arg1 ceca 799:802 pyloric ceca 791:802 pyloric ceca 791:802 Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. 26066491 5 4 theme acclimated 648:657 arg1 salmon 668:673 five freshwater acclimated Atlantic salmon 632:673 five freshwater acclimated Atlantic salmon 632:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 7 5 theme Skin 840:843 arg1 O-glycans 845:853 Skin O-glycans 840:853 Skin O-glycans 840:853 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 6 6 gly sialylated 739:748 arg1 most 729:732 most 729:732 most 729:732 Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. 26066491 5 7 theme freshwater 637:646 arg1 salmon 668:673 five freshwater acclimated Atlantic salmon 632:673 five freshwater acclimated Atlantic salmon 632:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 4 8 theme O-glycosylation 557:571 arg1 knowledge 536:544 knowledge 536:544 knowledge of piscine O-glycosylation 536:571 However, knowledge of piscine O-glycosylation is very limited. 26066491 8 9 theme acids 1050:1054 arg1 cores 1003:1007 O-glycan cores 1, 2, 3, and 5 994:1022 O-glycan cores 1, 2, 3, and 5 994:1022 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 9 theme acids 1050:1054 arg1 types 1034:1038 three types 1028:1038 three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) 1028:1080 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 3 10 gly O-glycosylated 367:380 arg1 mucins 382:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 3 10 gly O-glycosylated 367:380 arg1 part 424:427 an important part 411:427 an important part of the defense against pathogens 411:460 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 3 11 theme lectin-like 505:515 arg1 adhesins 517:524 lectin-like adhesins 505:524 lectin-like adhesins 505:524 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 4 12 theme piscine 549:555 arg1 O-glycosylation 557:571 piscine O-glycosylation 549:571 piscine O-glycosylation 549:571 However, knowledge of piscine O-glycosylation is very limited. 26066491 8 13 theme O-glycan 994:1001 arg1 cores 1003:1007 O-glycan cores 1, 2, 3, and 5 994:1022 O-glycan cores 1, 2, 3, and 5 994:1022 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 3 14 theme host 485:488 arg1 surfaces 490:497 host surfaces 485:497 host surfaces using lectin-like adhesins 485:524 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 8 15 theme Skin 974:977 arg1 mucins 979:984 Skin mucins 974:984 Skin mucins 974:984 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 0 16 theme Intestinal 98:107 arg1 Mucins 109:114 Intestinal Mucins 98:114 Intestinal Mucins 98:114 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 0 17 contain Carries 16:22 arg1 Salmon 9:14 Atlantic Salmon 0:14 Atlantic Salmon 0:14 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 0 17 contain Carries 16:22 arg2 Range 26:30 a Range 24:30 a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins 24:114 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 10 18 theme intervention 1454:1465 arg1 therapies 1467:1475 disease intervention therapies 1446:1475 disease intervention therapies 1446:1475 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 8 19 dep acids 1050:1054 arg1 Neu5Gc 1065:1070 Neu5Gc 1065:1070 Neu5Gc 1065:1070 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 19 dep acids 1050:1054 arg1 Kdn 1077:1079 Kdn 1077:1079 Kdn 1077:1079 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 19 dep acids 1050:1054 arg1 Neu5Ac 1057:1062 Neu5Ac 1057:1062 Neu5Ac 1057:1062 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 19 dep acids 1050:1054 arg1 acids 1050:1054 sialic acids 1043:1054 sialic acids (Neu5Ac, Neu5Gc, and Kdn) 1043:1080 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 5 20 theme mucin 607:611 arg1 O-glycosylation 613:627 mucin O-glycosylation 607:627 mucin O-glycosylation of five freshwater acclimated Atlantic salmon 607:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 10 21 theme importance 1360:1369 arg1 structures 1337:1346 structures 1337:1346 structures of putative importance 1337:1369 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 8 22 contain carried 986:992 arg2 cores 1003:1007 O-glycan cores 1, 2, 3, and 5 994:1022 O-glycan cores 1, 2, 3, and 5 994:1022 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 22 contain carried 986:992 arg1 mucins 979:984 Skin mucins 974:984 Skin mucins 974:984 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 22 contain carried 986:992 arg2 types 1034:1038 three types 1028:1038 three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) 1028:1080 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 0 23 theme Atlantic 0:7 arg1 Salmon 9:14 Atlantic Salmon 0:14 Atlantic Salmon 0:14 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 10 24 theme structural 1281:1290 arg1 characterization 1292:1307 This structural characterization 1276:1307 This structural characterization 1276:1307 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 8 25 theme predominant 1107:1117 arg1 structure 1119:1127 the predominant structure 1103:1127 the predominant structure 1103:1127 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 25 theme predominant 1107:1117 arg1 sialyl-Tn 1090:1098 sialyl-Tn 1090:1098 sialyl-Tn 1090:1098 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 10 26 theme disease 1446:1452 arg1 therapies 1467:1475 disease intervention therapies 1446:1475 disease intervention therapies 1446:1475 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 5 27 theme mass 682:685 arg1 spectrometry 687:698 mass spectrometry 682:698 mass spectrometry 682:698 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 7 28 dep shorter 860:866 arg1 residues 873:880 2-6 residues 869:880 2-6 residues 869:880 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 7 29 dep O-glycans 932:940 arg1 residues 948:955 2-13 residues 943:955 2-13 residues 943:955 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 7 29 dep O-glycans 932:940 arg1 structures 961:970 93 structures 958:970 93 structures 958:970 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 1 30 theme host-pathogen 190:202 arg1 interactions 204:215 host-pathogen interactions 190:215 host-pathogen interactions in fish 190:223 Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. 26066491 1 31 dep industry 142:149 arg1 increasing 152:161 increasing 152:161 increasing the need for understanding host-pathogen interactions in fish 152:223 Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. 26066491 2 32 theme mucus 270:274 arg1 layer 276:280 a mucus layer 268:280 a mucus layer composed of mucins 268:299 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 10 33 used used 1316:1319 arg2 characterization 1292:1307 This structural characterization 1276:1307 This structural characterization 1276:1307 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 9 34 dep cores 1161:1165 arg1 core 1236:1239 sialylated core 5 1225:1241 sialylated core 5 1225:1241 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 34 dep cores 1161:1165 arg1 structure 1265:1273 the most dominant structure 1247:1273 the most dominant structure 1247:1273 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 34 dep cores 1161:1165 arg1 present 1212:1218 present 1212:1218 present 1212:1218 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 5 35 gly O-glycosylation 613:627 arg1 salmon 668:673 five freshwater acclimated Atlantic salmon 632:673 five freshwater acclimated Atlantic salmon 632:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 8 36 contain had 1086:1088 arg2 sialyl-Tn 1090:1098 sialyl-Tn 1090:1098 sialyl-Tn 1090:1098 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 36 contain had 1086:1088 arg1 mucins 979:984 Skin mucins 974:984 Skin mucins 974:984 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 8 36 contain had 1086:1088 arg2 structure 1119:1127 the predominant structure 1103:1127 the predominant structure 1103:1127 Skin mucins carried O-glycan cores 1, 2, 3, and 5 and three types of sialic acids (Neu5Ac, Neu5Gc, and Kdn) and had sialyl-Tn as the predominant structure. 26066491 3 37 theme important 414:422 arg1 part 424:427 an important part 411:427 an important part of the defense against pathogens 411:460 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 3 37 theme important 414:422 arg1 mucins 382:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 9 38 theme dominant 1256:1263 arg1 core 1236:1239 sialylated core 5 1225:1241 sialylated core 5 1225:1241 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 38 theme dominant 1256:1263 arg1 structure 1265:1273 the most dominant structure 1247:1273 the most dominant structure 1247:1273 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 6 39 theme distal 822:827 arg1 intestine 829:837 proximal and distal intestine 809:837 proximal and distal intestine 809:837 Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. 26066491 3 40 theme O-glycosylated 367:380 arg1 mucins 382:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 3 40 theme O-glycosylated 367:380 arg1 part 424:427 an important part 411:427 an important part of the defense against pathogens 411:460 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 9 41 theme sialic 1200:1205 arg1 acid 1207:1210 the only sialic acid 1191:1210 the only sialic acid 1191:1210 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 5 42 theme Atlantic 659:666 arg1 salmon 668:673 five freshwater acclimated Atlantic salmon 632:673 five freshwater acclimated Atlantic salmon 632:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 0 43 theme Structures 50:59 arg1 Range 26:30 a Range 24:30 a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins 24:114 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 0 44 theme Novel 35:39 arg1 Structures 50:59 Novel O-Glycan Structures 35:59 Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins 35:114 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 5 45 theme salmon 668:673 arg1 O-glycosylation 613:627 mucin O-glycosylation 607:627 mucin O-glycosylation of five freshwater acclimated Atlantic salmon 607:673 We characterized mucin O-glycosylation of five freshwater acclimated Atlantic salmon, using mass spectrometry. 26066491 6 46 theme proximal 809:816 arg1 intestine 829:837 proximal and distal intestine 809:837 proximal and distal intestine 809:837 Of the 109 O-glycans found, most were sialylated and differed in distribution among skin, pyloric ceca, and proximal and distal intestine. 26066491 9 47 gly sialylated 1225:1234 arg1 core 1236:1239 sialylated core 5 1225:1241 sialylated core 5 1225:1241 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 47 gly sialylated 1225:1234 arg1 structure 1265:1273 the most dominant structure 1247:1273 the most dominant structure 1247:1273 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 2 48 theme first 309:313 arg1 skin 230:233 The skin 226:233 The skin 226:233 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 48 theme first 309:313 arg1 surfaces 247:254 mucosal surfaces 239:254 mucosal surfaces 239:254 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 48 theme first 309:313 arg1 point 315:319 the first point 305:319 the first point of contact between fish and pathogens 305:357 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 10 49 theme putative 1351:1358 arg1 importance 1360:1369 putative importance 1351:1369 putative importance 1351:1369 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 9 50 theme Intestinal 1130:1139 arg1 mucins 1141:1146 Intestinal mucins 1130:1146 Intestinal mucins 1130:1146 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 1 51 from interactions 204:215 arg1 fish 220:223 fish 220:223 fish 220:223 Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. 26066491 3 52 theme defense 436:442 arg1 part 424:427 an important part 411:427 an important part of the defense against pathogens 411:460 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 3 52 theme defense 436:442 arg1 mucins 382:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins 360:387 Highly O-glycosylated mucins have been shown to be an important part of the defense against pathogens, and pathogens bind to host surfaces using lectin-like adhesins. 26066491 10 53 from testing 1413:1419 arg1 therapies 1467:1475 disease intervention therapies 1446:1475 disease intervention therapies 1446:1475 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 10 53 from testing 1413:1419 arg1 assays 1435:1440 biological assays 1424:1440 biological assays 1424:1440 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 10 54 theme further 1405:1411 arg1 testing 1413:1419 further testing 1405:1419 further testing in biological assays and disease intervention therapies 1405:1475 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26066491 1 55 theme growing 134:140 arg1 industry 142:149 a growing industry 132:149 a growing industry 132:149 Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. 26066491 1 55 theme growing 134:140 arg1 Aquaculture 117:127 Aquaculture 117:127 Aquaculture 117:127 Aquaculture is a growing industry, increasing the need for understanding host-pathogen interactions in fish. 26066491 9 56 theme sialylated 1225:1234 arg1 core 1236:1239 sialylated core 5 1225:1241 sialylated core 5 1225:1241 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 56 theme sialylated 1225:1234 arg1 structure 1265:1273 the most dominant structure 1247:1273 the most dominant structure 1247:1273 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 0 57 theme O-Glycan 41:48 arg1 Structures 50:59 Novel O-Glycan Structures 35:59 Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins 35:114 Atlantic Salmon Carries a Range of Novel O-Glycan Structures Differentially Localized on Skin and Intestinal Mucins. 26066491 7 58 dep diverse 892:898 arg1 structures 904:913 33 structures 901:913 33 structures 901:913 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 2 59 theme mucosal 239:245 arg1 skin 230:233 The skin 226:233 The skin 226:233 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 59 theme mucosal 239:245 arg1 surfaces 247:254 mucosal surfaces 239:254 mucosal surfaces 239:254 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 2 59 theme mucosal 239:245 arg1 point 315:319 the first point 305:319 the first point of contact between fish and pathogens 305:357 The skin and mucosal surfaces, covered by a mucus layer composed of mucins, is the first point of contact between fish and pathogens. 26066491 7 60 theme intestinal 921:930 arg1 O-glycans 932:940 intestinal O-glycans 921:940 intestinal O-glycans (2-13 residues, 93 structures) 921:971 Skin O-glycans were shorter (2-6 residues) and less diverse (33 structures) than intestinal O-glycans (2-13 residues, 93 structures). 26066491 9 61 contain carried 1148:1154 arg1 mucins 1141:1146 Intestinal mucins 1130:1146 Intestinal mucins 1130:1146 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 9 61 contain carried 1148:1154 arg2 cores 1161:1165 cores 1, 2, and 5 1161:1177 cores 1, 2, and 5 1161:1177 Intestinal mucins carried only cores 1, 2, and 5, Neu5Ac was the only sialic acid present, and sialylated core 5 was the most dominant structure. 26066491 10 62 theme biological 1424:1433 arg1 assays 1435:1440 biological assays 1424:1440 biological assays 1424:1440 This structural characterization can be used for identifying structures of putative importance in host-pathogen interactions for further testing in biological assays and disease intervention therapies. 26241388 7 0 theme S2 1172:1173 arg1 derivative 1180:1189 the new FDL-deficient S2 cell derivative 1150:1189 the new FDL-deficient S2 cell derivative produced in this study 1150:1212 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 4 1 from N-glycans 768:776 arg1 cells 805:809 S2 cells 802:809 S2 cells 802:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 2 2 theme N-glycosylation 330:344 arg1 patterns 346:353 the N-glycosylation patterns 326:353 the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells 326:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 2 3 theme insect 384:389 arg1 cells 405:409 insect and mammalian cells 384:409 insect and mammalian cells 384:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 1 4 theme key 145:147 arg1 reaction 158:165 a key trimming reaction 143:165 a key trimming reaction 143:165 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 4 5 theme high 547:550 arg1 frequency 552:560 a high frequency 545:560 a high frequency of site-specific nucleotide insertions and deletions 545:613 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 6 6 theme FDL 1045:1047 arg1 role 1037:1040 the key role 1029:1040 the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways 1029:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 7 7 theme recombinant 1335:1345 arg1 glycoproteins 1347:1359 recombinant glycoproteins 1335:1359 recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1335:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 3 8 from gene 474:477 arg1 cells 509:513 Drosophila melanogaster S2 cells 482:513 Drosophila melanogaster S2 cells 482:513 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 4 9 theme insect-type 642:652 arg1 Man3GlcNAc2 681:691 Man3GlcNAc2 681:691 Man3GlcNAc2 681:691 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 4 9 theme insect-type 642:652 arg1 products 671:678 insect-type, paucimannosidic products 642:678 insect-type, paucimannosidic products (Man3GlcNAc2) 642:692 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 4 10 theme site-specific 565:577 arg1 insertions 590:599 site-specific nucleotide insertions 565:599 site-specific nucleotide insertions 565:599 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 5 11 theme insect 912:917 arg1 genes 924:928 insect cell genes 912:928 insect cell genes 912:928 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 6 12 theme mammalian 1101:1109 arg1 pathways 1131:1138 insect and mammalian N-glycan processing pathways 1090:1138 insect and mammalian N-glycan processing pathways 1090:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 4 13 theme elongated 734:742 arg1 GlcNAc2Man3GlcNAc2 779:796 GlcNAc2Man3GlcNAc2 779:796 GlcNAc2Man3GlcNAc2 779:796 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 4 13 theme elongated 734:742 arg1 N-glycans 768:776 partially elongated, mammalian-type complex N-glycans 724:776 partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells 724:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 3 14 theme fdl 470:472 arg1 gene 474:477 the fdl gene 466:477 the fdl gene in Drosophila melanogaster S2 cells 466:513 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 6 15 theme major 1056:1060 arg1 point 1069:1073 the major branch point 1052:1073 the major branch point distinguishing insect and mammalian N-glycan processing pathways 1052:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 7 16 theme bottom-up 1233:1241 arg1 efforts 1260:1266 future bottom-up glycoengineering efforts 1226:1266 future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1226:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 5 17 theme cell 919:922 arg1 genes 924:928 insect cell genes 912:928 insect cell genes 912:928 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 2 18 from differences 311:321 arg1 patterns 346:353 the N-glycosylation patterns 326:353 the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells 326:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 4 19 theme complex 760:766 arg1 GlcNAc2Man3GlcNAc2 779:796 GlcNAc2Man3GlcNAc2 779:796 GlcNAc2Man3GlcNAc2 779:796 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 4 19 theme complex 760:766 arg1 N-glycans 768:776 partially elongated, mammalian-type complex N-glycans 724:776 partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells 724:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 1 20 theme trimming 149:156 arg1 reaction 158:165 a key trimming reaction 143:165 a key trimming reaction 143:165 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 7 21 gly glycoproteins 1347:1359 arg1 glycoproteins 1347:1359 recombinant glycoproteins 1335:1359 recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1335:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 5 22 theme protein 873:879 arg1 pathways 895:902 protein glycosylation pathways 873:902 protein glycosylation pathways 873:902 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 6 23 theme branch 1062:1067 arg1 point 1069:1073 the major branch point 1052:1073 the major branch point distinguishing insect and mammalian N-glycan processing pathways 1052:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 1 24 theme Fused 82:86 arg1 enzyme 106:111 an enzyme 103:111 an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway 103:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 1 24 theme Fused 82:86 arg1 FDL 95:97 FDL 95:97 FDL 95:97 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 1 24 theme Fused 82:86 arg1 lobes 88:92 Fused lobes 82:92 Fused lobes (FDL) 82:98 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 6 25 theme key 1033:1035 arg1 role 1037:1040 the key role 1029:1040 the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways 1029:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 6 26 theme N-glycan 1111:1118 arg1 pathways 1131:1138 insect and mammalian N-glycan processing pathways 1090:1138 insect and mammalian N-glycan processing pathways 1090:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 2 27 gly glycoproteins 358:370 arg1 glycoproteins 358:370 glycoproteins 358:370 glycoproteins produced by insect and mammalian cells 358:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 3 28 used used 430:433 arg2 we 427:428 we 427:428 we 427:428 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 7 29 theme future 1226:1231 arg1 efforts 1260:1266 future bottom-up glycoengineering efforts 1226:1266 future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1226:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 7 30 theme insect 1288:1293 arg1 lines 1300:1304 insect cell lines 1288:1304 insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1288:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 4 31 from production 710:719 arg1 cells 805:809 S2 cells 802:809 S2 cells 802:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 5 32 used used 847:850 arg2 CRISPR-Cas9 815:825 CRISPR-Cas9 815:825 CRISPR-Cas9 815:825 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 2 33 theme glycoproteins 358:370 arg1 patterns 346:353 the N-glycosylation patterns 326:353 the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells 326:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 0 34 theme Cell 20:23 arg1 Pathway 45:51 an Insect Cell N-Glycan Processing Pathway 10:51 an Insect Cell N-Glycan Processing Pathway 10:51 Modifying an Insect Cell N-Glycan Processing Pathway Using CRISPR-Cas Technology. 26241388 4 35 theme CRISPR-Cas9 516:526 arg1 editing 528:534 CRISPR-Cas9 editing 516:534 CRISPR-Cas9 editing 516:534 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 7 36 theme oligosaccharide 1388:1402 arg1 structures 1415:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 1 37 theme elongation 183:192 arg1 reactions 194:202 elongation reactions 183:202 elongation reactions in the insect N-glycan processing pathway 183:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 0 38 theme Insect 13:18 arg1 Pathway 45:51 an Insect Cell N-Glycan Processing Pathway 10:51 an Insect Cell N-Glycan Processing Pathway 10:51 Modifying an Insect Cell N-Glycan Processing Pathway Using CRISPR-Cas Technology. 26241388 4 39 theme insertions 590:599 arg1 frequency 552:560 a high frequency 545:560 a high frequency of site-specific nucleotide insertions and deletions 545:613 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 3 40 dep Drosophila 482:491 arg1 melanogaster 493:504 melanogaster 493:504 melanogaster 493:504 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 3 41 theme CRISPR-Cas9 439:449 arg1 system 451:456 the CRISPR-Cas9 system 435:456 the CRISPR-Cas9 system 435:456 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 0 42 theme Processing 34:43 arg1 Pathway 45:51 an Insect Cell N-Glycan Processing Pathway 10:51 an Insect Cell N-Glycan Processing Pathway 10:51 Modifying an Insect Cell N-Glycan Processing Pathway Using CRISPR-Cas Technology. 26241388 4 43 theme S2 802:803 arg1 cells 805:809 S2 cells 802:809 S2 cells 802:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 7 44 theme cell 1295:1298 arg1 lines 1300:1304 insect cell lines 1288:1304 insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1288:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 2 45 theme mammalian 395:403 arg1 cells 405:409 insect and mammalian cells 384:409 insect and mammalian cells 384:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 0 46 theme N-Glycan 25:32 arg1 Pathway 45:51 an Insect Cell N-Glycan Processing Pathway 10:51 an Insect Cell N-Glycan Processing Pathway 10:51 Modifying an Insect Cell N-Glycan Processing Pathway Using CRISPR-Cas Technology. 26241388 7 47 theme cell 1175:1178 arg1 derivative 1180:1189 the new FDL-deficient S2 cell derivative 1150:1189 the new FDL-deficient S2 cell derivative produced in this study 1150:1212 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 2 48 gly N-glycosylation 330:344 arg1 glycoproteins 358:370 glycoproteins 358:370 glycoproteins produced by insect and mammalian cells 358:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 4 49 theme deletions 605:613 arg1 frequency 552:560 a high frequency 545:560 a high frequency of site-specific nucleotide insertions and deletions 545:613 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 7 50 theme new 1154:1156 arg1 derivative 1180:1189 the new FDL-deficient S2 cell derivative 1150:1189 the new FDL-deficient S2 cell derivative produced in this study 1150:1212 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 7 51 with glycoproteins 1347:1359 arg1 structures 1415:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 7 52 theme predefined 1377:1386 arg1 structures 1415:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 4 53 from cells 805:809 arg1 production 710:719 the production 706:719 the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells 706:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 5 54 theme glycosylation 881:893 arg1 pathways 895:902 protein glycosylation pathways 873:902 protein glycosylation pathways 873:902 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 4 55 theme nucleotide 579:588 arg1 insertions 590:599 site-specific nucleotide insertions 565:599 site-specific nucleotide insertions 565:599 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 6 56 from point 1069:1073 arg1 role 1037:1040 the key role 1029:1040 the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways 1029:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 7 57 theme glycoengineering 1243:1258 arg1 efforts 1260:1266 future bottom-up glycoengineering efforts 1226:1266 future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures 1226:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 4 58 dep elongated 734:742 arg1 mammalian-type 745:758 mammalian-type 745:758 mammalian-type 745:758 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 5 59 theme broad 960:964 arg1 utility 966:972 its broad utility 956:972 its broad utility 956:972 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 5 59 theme broad 960:964 arg1 tool 979:982 a tool 977:982 a tool for these purposes 977:1001 As CRISPR-Cas9 has not been widely used to analyze or modify protein glycosylation pathways or edit insect cell genes, these results underscore its broad utility as a tool for these purposes. 26241388 2 60 theme FDL 260:262 arg1 function 264:271 FDL function 260:271 FDL function 260:271 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 1 61 from reactions 194:202 arg1 pathway 238:244 the insect N-glycan processing pathway 207:244 the insect N-glycan processing pathway 207:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 2 62 theme major 305:309 arg1 differences 311:321 major differences 305:321 major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells 305:409 Accordingly, FDL function accounts, at least in part, for major differences in the N-glycosylation patterns of glycoproteins produced by insect and mammalian cells. 26241388 1 63 theme insect 211:216 arg1 pathway 238:244 the insect N-glycan processing pathway 207:244 the insect N-glycan processing pathway 207:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 3 64 theme S2 506:507 arg1 cells 509:513 Drosophila melanogaster S2 cells 482:513 Drosophila melanogaster S2 cells 482:513 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 4 65 theme N-glycans 768:776 arg1 production 710:719 the production 706:719 the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells 706:809 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 4 66 theme products 671:678 arg1 production 628:637 the production 624:637 the production of insect-type, paucimannosidic products (Man3GlcNAc2) 624:692 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 1 67 theme N-glycan 218:225 arg1 pathway 238:244 the insect N-glycan processing pathway 207:244 the insect N-glycan processing pathway 207:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 4 68 dep insect-type 642:652 arg1 paucimannosidic 655:669 paucimannosidic 655:669 paucimannosidic 655:669 CRISPR-Cas9 editing produced a high frequency of site-specific nucleotide insertions and deletions, reduced the production of insect-type, paucimannosidic products (Man3GlcNAc2), and led to the production of partially elongated, mammalian-type complex N-glycans (GlcNAc2Man3GlcNAc2) in S2 cells. 26241388 6 69 theme processing 1120:1129 arg1 pathways 1131:1138 insect and mammalian N-glycan processing pathways 1090:1138 insect and mammalian N-glycan processing pathways 1090:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 1 70 theme processing 227:236 arg1 pathway 238:244 the insect N-glycan processing pathway 207:244 the insect N-glycan processing pathway 207:244 Fused lobes (FDL) is an enzyme that simultaneously catalyzes a key trimming reaction and antagonizes elongation reactions in the insect N-glycan processing pathway. 26241388 0 71 theme CRISPR-Cas 59:68 arg1 Technology 70:79 CRISPR-Cas Technology 59:79 CRISPR-Cas Technology 59:79 Modifying an Insect Cell N-Glycan Processing Pathway Using CRISPR-Cas Technology. 26241388 6 72 theme insect 1090:1095 arg1 pathways 1131:1138 insect and mammalian N-glycan processing pathways 1090:1138 insect and mammalian N-glycan processing pathways 1090:1138 Our results also confirm the key role of FDL at the major branch point distinguishing insect and mammalian N-glycan processing pathways. 26241388 3 73 theme Drosophila 482:491 arg1 cells 509:513 Drosophila melanogaster S2 cells 482:513 Drosophila melanogaster S2 cells 482:513 In this study, we used the CRISPR-Cas9 system to edit the fdl gene in Drosophila melanogaster S2 cells. 26241388 7 74 theme side-chain 1404:1413 arg1 structures 1415:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 chemically predefined oligosaccharide side-chain structures 1366:1424 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 26241388 7 75 theme FDL-deficient 1158:1170 arg1 derivative 1180:1189 the new FDL-deficient S2 cell derivative 1150:1189 the new FDL-deficient S2 cell derivative produced in this study 1150:1212 Finally, the new FDL-deficient S2 cell derivative produced in this study will enable future bottom-up glycoengineering efforts designed to isolate insect cell lines that can efficiently produce recombinant glycoproteins with chemically predefined oligosaccharide side-chain structures. 28417175 1 0 theme tris 302:305 arg1 acid 349:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid 302:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 4 1 theme test 944:947 arg1 value 951:955 T test p value <0.01 942:961 T test p value <0.01 942:961 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 1 theme test 944:947 arg1 differences 929:939 significant differences 917:939 significant differences (T test p value <0.01) of 6 glycan structures 917:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 1 2 theme 2,4,6-trimethoxyphenyl 307:328 arg1 acid 349:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid 302:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 5 3 from controls 1210:1217 arg1 expression 1222:1231 expression 1222:1231 expression of the selected glycans 1222:1255 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 0 4 theme Microwave-assisted 0:17 arg1 deglycosylation 19:33 Microwave-assisted deglycosylation 0:33 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 0:113 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 28417175 4 5 theme significant 917:927 arg1 result 909:914 a result 907:914 a result 907:914 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 5 theme significant 917:927 arg1 differences 929:939 significant differences 917:939 significant differences (T test p value <0.01) of 6 glycan structures 917:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 5 theme significant 917:927 arg1 value 951:955 T test p value <0.01 942:961 T test p value <0.01 942:961 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 2 6 theme MALDI-MS 600:607 arg1 sensitivity 609:619 MALDI-MS sensitivity 600:619 MALDI-MS sensitivity 600:619 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 3 7 theme human 796:800 arg1 serum 802:806 human serum 796:806 human serum 796:806 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 6 8 theme potential 1345:1353 arg1 biomarkers 1380:1389 potential lung cancer glycan-based biomarkers 1345:1389 potential lung cancer glycan-based biomarkers 1345:1389 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 8 theme potential 1345:1353 arg1 structures 1319:1328 these structures 1313:1328 these structures 1313:1328 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 4 9 theme structures 976:985 arg1 result 909:914 a result 907:914 a result 907:914 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 9 theme structures 976:985 arg1 differences 929:939 significant differences 917:939 significant differences (T test p value <0.01) of 6 glycan structures 917:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 9 theme structures 976:985 arg1 value 951:955 T test p value <0.01 942:961 T test p value <0.01 942:961 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 10 theme sample 1143:1148 arg1 sets 1150:1153 two sample sets 1139:1153 two sample sets 1139:1153 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 6 11 attach present 1529:1535 arg2 N-glycome 1519:1527 N-glycome 1519:1527 N-glycome present in human serum 1519:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 11 attach present 1529:1535 arg1 serum 1546:1550 human serum 1540:1550 human serum 1540:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 1 12 theme phosphonium 330:340 arg1 acid 349:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid 302:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 3 13 theme pathological 824:835 arg1 analysis 837:844 preliminary pathological analysis 812:844 preliminary pathological analysis of N-glycans between healthy and lung cancer individuals 812:901 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 1 14 theme N-glycan 137:144 arg1 profiling 146:154 A rapid and sensitive N-glycan profiling 115:154 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 115:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 3 15 theme other 695:699 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 15 theme other 695:699 arg1 ovalbumin 729:737 ovalbumin 729:737 ovalbumin 729:737 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 15 theme other 695:699 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 4 16 theme N-glycan 1020:1027 arg1 structures 1029:1038 54 detected N-glycan structures 1008:1038 54 detected N-glycan structures with only 50 nL of loading amount 1008:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 17 with validation 679:688 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 17 with validation 679:688 arg1 ovalbumin 729:737 ovalbumin 729:737 ovalbumin 729:737 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 17 with validation 679:688 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 5 18 theme T 1278:1278 arg1 test 1280:1283 T test 1278:1283 T test 1278:1283 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 0 19 theme glycosylamine 85:97 arg1 derivatization 99:112 glycosylamine derivatization 85:112 glycosylamine derivatization 85:112 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 28417175 4 20 theme T 942:942 arg1 value 951:955 T test p value <0.01 942:961 T test p value <0.01 942:961 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 20 theme T 942:942 arg1 differences 929:939 significant differences 917:939 significant differences (T test p value <0.01) of 6 glycan structures 917:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 7 21 theme microwave-assisted 1610:1627 arg1 deglycosylation 1629:1643 microwave-assisted deglycosylation 1610:1643 microwave-assisted deglycosylation 1610:1643 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 3 22 theme cancer 884:889 arg1 individuals 891:901 healthy and lung cancer individuals 867:901 healthy and lung cancer individuals 867:901 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 7 23 theme N-glycans 1597:1605 arg1 analysis 1585:1592 Graphical abstract MALDI-TOF MS analysis 1553:1592 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 1553:1677 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 6 24 theme cancer 1360:1365 arg1 biomarkers 1380:1389 potential lung cancer glycan-based biomarkers 1345:1389 potential lung cancer glycan-based biomarkers 1345:1389 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 24 theme cancer 1360:1365 arg1 structures 1319:1328 these structures 1313:1328 these structures 1313:1328 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 25 theme proposed 1468:1475 arg1 method 1477:1482 proposed method 1468:1482 proposed method 1468:1482 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 5 26 theme cancer 1193:1198 arg1 stage 1200:1204 each lung cancer stage 1183:1204 each lung cancer stage 1183:1204 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 1 27 theme acetic 342:347 arg1 acid 349:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid 302:352 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 7 28 theme MS 1582:1583 arg1 analysis 1585:1592 Graphical abstract MALDI-TOF MS analysis 1553:1592 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 1553:1677 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 0 29 theme N-glycans 71:79 arg1 analysis 59:66 rapid and sensitive analysis 39:66 rapid and sensitive analysis of N-glycans 39:79 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 28417175 2 30 theme efficient 464:472 arg1 release 474:480 highly efficient release 457:480 highly efficient release 457:480 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 1 31 theme N-hydroxysuccinimide 354:373 arg1 ester 375:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester 302:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 31 theme N-hydroxysuccinimide 354:373 arg1 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 7 32 theme abstract 1563:1570 arg1 analysis 1585:1592 Graphical abstract MALDI-TOF MS analysis 1553:1592 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 1553:1677 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 4 33 theme loading 1059:1065 arg1 amount 1067:1072 loading amount 1059:1072 loading amount 1059:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 34 theme standard 705:712 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 34 theme standard 705:712 arg1 ovalbumin 729:737 ovalbumin 729:737 ovalbumin 729:737 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 34 theme standard 705:712 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 6 35 theme cancer 1421:1426 arg1 progression 1428:1438 cancer progression 1421:1438 cancer progression 1421:1438 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 36 from present 1529:1535 arg1 serum 1546:1550 human serum 1540:1550 human serum 1540:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 37 theme present 1529:1535 arg1 N-glycome 1519:1527 N-glycome 1519:1527 N-glycome present in human serum 1519:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 5 38 theme selected 1240:1247 arg1 glycans 1249:1255 the selected glycans 1236:1255 the selected glycans 1236:1255 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 3 39 dep glycoproteins 714:726 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 39 dep glycoproteins 714:726 arg1 ovalbumin 729:737 ovalbumin 729:737 ovalbumin 729:737 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 39 dep glycoproteins 714:726 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 4 40 theme PCA 1104:1106 arg1 analyses 1122:1129 PCA and ROC (AUC) analyses 1104:1129 PCA and ROC (AUC) analyses between two sample sets 1104:1153 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 41 theme proposed 763:770 arg1 strategy 772:779 the proposed strategy 759:779 the proposed strategy 759:779 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 4 42 theme ROC 1112:1114 arg1 analyses 1122:1129 PCA and ROC (AUC) analyses 1104:1129 PCA and ROC (AUC) analyses between two sample sets 1104:1153 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 43 gly glycoproteins 714:726 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 43 gly glycoproteins 714:726 arg1 ovalbumin 729:737 ovalbumin 729:737 ovalbumin 729:737 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 43 gly glycoproteins 714:726 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 5 44 from trend 1174:1178 arg1 expression 1222:1231 expression 1222:1231 expression of the selected glycans 1222:1255 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 2 45 dep 35-fold 577:583 arg1 to 574:575 to 574:575 to 574:575 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 2 46 theme underivatized 640:652 arg1 N-glycans 654:662 underivatized N-glycans 640:662 underivatized N-glycans 640:662 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 2 47 theme ribonuclease 512:523 arg1 B 525:525 ribonuclease B 512:525 ribonuclease B 512:525 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 1 48 theme deglycosylation 203:217 arg1 use 196:198 the use 192:198 the use of deglycosylation with microwave assistance 192:243 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 48 theme deglycosylation 203:217 arg1 co-derivatization 253:269 the co-derivatization 249:269 the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 249:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 4 49 theme amount 1067:1072 arg1 50 nL 1050:1054 only 50 nL 1045:1054 only 50 nL of loading amount 1045:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 50 theme preliminary 812:822 arg1 analysis 837:844 preliminary pathological analysis 812:844 preliminary pathological analysis of N-glycans between healthy and lung cancer individuals 812:901 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 6 51 theme lung 1355:1358 arg1 biomarkers 1380:1389 potential lung cancer glycan-based biomarkers 1345:1389 potential lung cancer glycan-based biomarkers 1345:1389 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 51 theme lung 1355:1358 arg1 structures 1319:1328 these structures 1313:1328 these structures 1313:1328 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 1 52 theme microwave 224:232 arg1 assistance 234:243 microwave assistance 224:243 microwave assistance 224:243 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 6 53 theme human 1540:1544 arg1 serum 1546:1550 human serum 1540:1550 human serum 1540:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 54 used used 1337:1340 arg2 structures 1319:1328 these structures 1313:1328 these structures 1313:1328 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 54 used used 1337:1340 arg2 biomarkers 1380:1389 potential lung cancer glycan-based biomarkers 1345:1389 potential lung cancer glycan-based biomarkers 1345:1389 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 4 55 theme glycan 969:974 arg1 structures 976:985 6 glycan structures 967:985 6 glycan structures 967:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 56 theme N-glycans 849:857 arg1 analysis 837:844 preliminary pathological analysis 812:844 preliminary pathological analysis of N-glycans between healthy and lung cancer individuals 812:901 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 1 57 theme sensitive 127:135 arg1 profiling 146:154 A rapid and sensitive N-glycan profiling 115:154 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 115:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 4 58 theme p 949:949 arg1 value 951:955 T test p value <0.01 942:961 T test p value <0.01 942:961 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 4 58 theme p 949:949 arg1 differences 929:939 significant differences 917:939 significant differences (T test p value <0.01) of 6 glycan structures 917:985 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 1 59 theme profiling 146:154 arg1 strategy 156:163 A rapid and sensitive N-glycan profiling strategy 115:163 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 115:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 7 60 theme glycosylamine 1649:1661 arg1 derivatization 1663:1676 glycosylamine derivatization 1649:1676 glycosylamine derivatization 1649:1676 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 2 61 theme sensitivity 609:619 arg1 enhancement 585:595 up to 35-fold enhancement 571:595 up to 35-fold enhancement of MALDI-MS sensitivity 571:619 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 3 62 theme lung 879:882 arg1 cancer 884:889 lung cancer 879:889 lung cancer 879:889 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 63 theme further 671:677 arg1 validation 679:688 further validation 671:688 further validation with other two standard glycoproteins (ovalbumin and bovine fetuin) 671:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 4 64 theme detected 1011:1018 arg1 structures 1029:1038 54 detected N-glycan structures 1008:1038 54 detected N-glycan structures with only 50 nL of loading amount 1008:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 3 65 theme healthy 867:873 arg1 individuals 891:901 healthy and lung cancer individuals 867:901 healthy and lung cancer individuals 867:901 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 2 66 from B 525:525 arg1 tagging 486:492 tagging 486:492 tagging 486:492 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 2 66 from B 525:525 arg1 release 474:480 highly efficient release 457:480 highly efficient release 457:480 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 6 67 theme glycan-based 1367:1378 arg1 biomarkers 1380:1389 potential lung cancer glycan-based biomarkers 1345:1389 potential lung cancer glycan-based biomarkers 1345:1389 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 67 theme glycan-based 1367:1378 arg1 structures 1319:1328 these structures 1313:1328 these structures 1313:1328 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 68 theme method 1477:1482 arg1 ability 1457:1463 the ability 1453:1463 the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum 1453:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 6 69 from serum 1546:1550 arg1 present 1529:1535 present 1529:1535 present 1529:1535 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 2 70 theme N-glycans 497:505 arg1 tagging 486:492 tagging 486:492 tagging 486:492 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 2 70 theme N-glycans 497:505 arg1 release 474:480 highly efficient release 457:480 highly efficient release 457:480 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 0 71 theme rapid 39:43 arg1 analysis 59:66 rapid and sensitive analysis 39:66 rapid and sensitive analysis of N-glycans 39:79 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 28417175 5 72 theme stage 1200:1204 arg1 trend 1174:1178 the trend 1170:1178 the trend of each lung cancer stage 1170:1204 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 5 72 theme stage 1200:1204 arg1 controls 1210:1217 controls 1210:1217 controls in expression of the selected glycans 1210:1255 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 1 73 theme rapid 117:121 arg1 profiling 146:154 A rapid and sensitive N-glycan profiling 115:154 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 115:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 0 74 theme sensitive 49:57 arg1 analysis 59:66 rapid and sensitive analysis 39:66 rapid and sensitive analysis of N-glycans 39:79 Microwave-assisted deglycosylation for rapid and sensitive analysis of N-glycans via glycosylamine derivatization. 28417175 1 75 theme acid 349:352 arg1 ester 375:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester 302:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 75 theme acid 349:352 arg1 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 4 76 theme only 1045:1048 arg1 50 nL 1050:1054 only 50 nL 1045:1054 only 50 nL of loading amount 1045:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 1 77 theme glycosylamine 274:286 arg1 labeling 288:295 glycosylamine labeling 274:295 glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 274:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 6 78 theme progression 1428:1438 arg1 definition 1407:1416 further definition 1399:1416 further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum 1399:1550 Accordingly, these structures can be used as potential lung cancer glycan-based biomarkers and for further definition of cancer progression highlighting the ability of proposed method to rapidly and efficiently analyze N-glycome present in human serum. 28417175 1 79 with deglycosylation 203:217 arg1 assistance 234:243 microwave assistance 224:243 microwave assistance 224:243 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 7 80 theme MALDI-TOF 1572:1580 arg1 analysis 1585:1592 Graphical abstract MALDI-TOF MS analysis 1553:1592 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 1553:1677 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 4 81 with structures 1029:1038 arg1 50 nL 1050:1054 only 50 nL 1045:1054 only 50 nL of loading amount 1045:1072 As a result, significant differences (T test p value <0.01) of 6 glycan structures were determined from 54 detected N-glycan structures with only 50 nL of loading amount and further confirmed through PCA and ROC (AUC) analyses between two sample sets. 28417175 2 82 theme 35-fold 577:583 arg1 enhancement 585:595 up to 35-fold enhancement 571:595 up to 35-fold enhancement of MALDI-MS sensitivity 571:619 Notably, highly efficient release and tagging of N-glycans from ribonuclease B was achieved in less than 90 min, providing up to 35-fold enhancement of MALDI-MS sensitivity with comparison to underivatized N-glycans. 28417175 3 83 theme bovine 743:748 arg1 glycoproteins 714:726 two standard glycoproteins 701:726 other two standard glycoproteins (ovalbumin and bovine fetuin) 695:756 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 3 83 theme bovine 743:748 arg1 fetuin 750:755 bovine fetuin 743:755 bovine fetuin 743:755 After further validation with other two standard glycoproteins (ovalbumin and bovine fetuin), the proposed strategy was applied to human serum for preliminary pathological analysis of N-glycans between healthy and lung cancer individuals. 28417175 5 84 theme lung 1188:1191 arg1 stage 1200:1204 each lung cancer stage 1183:1204 each lung cancer stage 1183:1204 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 7 85 theme Graphical 1553:1561 arg1 analysis 1585:1592 Graphical abstract MALDI-TOF MS analysis 1553:1592 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 1553:1677 Graphical abstract MALDI-TOF MS analysis of N-glycans by microwave-assisted deglycosylation and glycosylamine derivatization. 28417175 5 86 theme glycans 1249:1255 arg1 expression 1222:1231 expression 1222:1231 expression of the selected glycans 1222:1255 Subsequently, the trend of each lung cancer stage and controls in expression of the selected glycans was implemented with T test and box-plots. 28417175 1 87 with labeling 288:295 arg1 methylamidation 399:413 methylamidation 399:413 methylamidation 399:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 87 with labeling 288:295 arg1 ester 375:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester 302:379 tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) 302:393 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 87 with labeling 288:295 arg1 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 TMPP-Ac-OSu 382:392 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 88 theme labeling 288:295 arg1 use 196:198 the use 192:198 the use of deglycosylation with microwave assistance 192:243 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 28417175 1 88 theme labeling 288:295 arg1 co-derivatization 253:269 the co-derivatization 249:269 the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation 249:413 A rapid and sensitive N-glycan profiling strategy for MALDI-MS incorporating the use of deglycosylation with microwave assistance and the co-derivatization of glycosylamine labeling with tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide ester (TMPP-Ac-OSu) and methylamidation has been developed in this work. 26925665 3 0 link O-linked 318:325 arg1 glycoprotein 327:338 O-linked glycoprotein 318:338 O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 318:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 1 from structures 304:313 arg1 lines 375:379 cholangiocarcinoma (CCA) cell lines 345:379 cholangiocarcinoma (CCA) cell lines from different histological types 345:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 2 2 theme protein 185:191 arg1 glycosylation 193:205 Aberrant protein glycosylation 176:205 Aberrant protein glycosylation 176:205 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 8 3 from role 1543:1546 arg1 cells 1574:1578 cancer cells 1567:1578 cancer cells 1567:1578 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 6 4 dep appeared 1275:1282 arg1 dominated 1284:1292 dominated 1284:1292 appeared dominated by NeuAc2Gal1GalNAc1 1275:1313 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 9 5 theme disease-related 1674:1688 arg1 glycoproteins 1690:1702 disease-related glycoproteins 1674:1702 disease-related glycoproteins 1674:1702 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 6 6 theme differentiated 1184:1197 arg1 % 1225:1225 M214; 40.1% 1215:1225 M214; 40.1% 1215:1225 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 6 theme differentiated 1184:1197 arg1 adenocarcinoma 1199:1212 moderately to poorly differentiated adenocarcinoma 1163:1212 moderately to poorly differentiated adenocarcinoma (M214; 40.1%) 1163:1226 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 5 7 theme cell 647:650 arg1 lines 652:656 the CCA cell lines 639:656 the CCA cell lines 639:656 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 7 8 theme histological 1407:1418 arg1 types 1420:1424 the different histological types 1393:1424 the different histological types of CCA 1393:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 5 9 theme sialic 721:726 arg1 acids 728:732 sialic acids 721:732 sialic acids 721:732 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 6 10 theme differentiated 1014:1027 arg1 % 1055:1055 K100; 57.1% 1045:1055 K100; 57.1% 1045:1055 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 10 theme differentiated 1014:1027 arg1 adenocarcinoma 1029:1042 poorly differentiated adenocarcinoma 1007:1042 poorly differentiated adenocarcinoma (K100; 57.1%) 1007:1056 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 3 11 theme glycan 297:302 arg1 structures 304:313 the glycan structures 293:313 the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 293:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 8 12 theme cell 1447:1450 arg1 lines 1452:1456 All five CCA cell lines 1434:1456 All five CCA cell lines 1434:1456 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 6 13 theme cell 1246:1249 arg1 % 1272:1272 M213; 34.7% 1262:1272 M213; 34.7% 1262:1272 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 13 theme cell 1246:1249 arg1 carcinoma 1251:1259 adenosquamous cell carcinoma 1232:1259 adenosquamous cell carcinoma (M213; 34.7%) 1232:1273 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 3 14 theme cholangiocarcinoma 345:362 arg1 lines 375:379 cholangiocarcinoma (CCA) cell lines 345:379 cholangiocarcinoma (CCA) cell lines from different histological types 345:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 7 15 theme glycans 1382:1388 arg1 expression 1355:1364 differential expression 1342:1364 differential expression of the O-linked glycans in the different histological types of CCA 1342:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 3 16 theme glycoprotein 327:338 arg1 structures 304:313 the glycan structures 293:313 the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 293:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 6 17 theme abundant 985:992 arg1 NeuAc1Gal1GalNAc1 945:961 NeuAc1Gal1GalNAc1 945:961 NeuAc1Gal1GalNAc1 945:961 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 17 theme abundant 985:992 arg1 structure 994:1002 the most abundant structure 976:1002 the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%) 976:1154 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 5 18 dep NeuAc1Gal1GalNAc1 735:751 arg1 lines 864:868 All five CCA cell lines 846:868 All five CCA cell lines 846:868 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 8 19 theme terminal 1472:1479 arg1 NeuAc 1494:1498 NeuAc 1494:1498 NeuAc 1494:1498 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 8 19 theme terminal 1472:1479 arg1 acid 1488:1491 terminal sialic acid 1472:1491 terminal sialic acid (NeuAc) 1472:1499 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 1 20 theme mammalian 159:167 arg1 cells 169:173 mammalian cells 159:173 mammalian cells 159:173 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 6 21 theme differentiated 1070:1083 arg1 % 1111:1111 M055; 42.6% 1101:1111 M055; 42.6% 1101:1111 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 21 theme differentiated 1070:1083 arg1 adenocarcinoma 1085:1098 moderately differentiated adenocarcinoma 1059:1098 moderately differentiated adenocarcinoma (M055; 42.6%) 1059:1112 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 3 22 theme cell 370:373 arg1 lines 375:379 cholangiocarcinoma (CCA) cell lines 345:379 cholangiocarcinoma (CCA) cell lines from different histological types 345:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 23 theme CCA 365:367 arg1 lines 375:379 cholangiocarcinoma (CCA) cell lines 345:379 cholangiocarcinoma (CCA) cell lines from different histological types 345:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 5 24 link O-linked 619:626 arg1 glycans 628:634 the O-linked glycans 615:634 the O-linked glycans of the CCA cell lines 615:656 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 3 25 theme histological 396:407 arg1 types 409:413 different histological types 386:413 different histological types 386:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 8 26 theme O-linked 1501:1508 arg1 glycans 1510:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 5 27 theme O-linked 619:626 arg1 glycans 628:634 the O-linked glycans 615:634 the O-linked glycans of the CCA cell lines 615:656 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 7 28 from expression 1355:1364 arg1 types 1420:1424 the different histological types 1393:1424 the different histological types of CCA 1393:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 4 29 theme human 517:521 arg1 K100 539:542 K100 539:542 K100 539:542 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 29 theme human 517:521 arg1 M055 545:548 M055 545:548 M055 545:548 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 29 theme human 517:521 arg1 M214 566:569 M214 566:569 M214 566:569 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 29 theme human 517:521 arg1 lines 532:536 Five human CCA cell lines 512:536 Five human CCA cell lines 512:536 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 29 theme human 517:521 arg1 M139 551:554 M139 551:554 M139 551:554 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 29 theme human 517:521 arg1 M213 557:560 M213 557:560 M213 557:560 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 7 30 theme differential 1342:1353 arg1 expression 1355:1364 differential expression 1342:1364 differential expression of the O-linked glycans in the different histological types of CCA 1342:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 4 31 theme cell 527:530 arg1 K100 539:542 K100 539:542 K100 539:542 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 31 theme cell 527:530 arg1 M055 545:548 M055 545:548 M055 545:548 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 31 theme cell 527:530 arg1 M214 566:569 M214 566:569 M214 566:569 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 31 theme cell 527:530 arg1 lines 532:536 Five human CCA cell lines 512:536 Five human CCA cell lines 512:536 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 31 theme cell 527:530 arg1 M139 551:554 M139 551:554 M139 551:554 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 31 theme cell 527:530 arg1 M213 557:560 M213 557:560 M213 557:560 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 3 32 theme trap 478:481 arg1 spectrometry 488:499 nanospray ionization-linear ion trap mass spectrometry 446:499 nanospray ionization-linear ion trap mass spectrometry (NSI-MSn) 446:509 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 32 theme trap 478:481 arg1 NSI-MSn 502:508 NSI-MSn 502:508 NSI-MSn 502:508 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 0 33 theme Differential 0:11 arg1 Expression 13:22 Differential Expression 0:22 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 0:82 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 5 34 theme similar 879:885 arg1 pattern 894:900 a similar glycan pattern 877:900 a similar glycan pattern 877:900 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 1 35 theme common 118:123 arg1 glycosylation 92:104 Protein glycosylation 84:104 Protein glycosylation 84:104 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 1 35 theme common 118:123 arg1 modification 143:154 the most common posttranslational modification 109:154 the most common posttranslational modification in mammalian cells 109:173 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 3 36 theme ionization-linear 456:472 arg1 spectrometry 488:499 nanospray ionization-linear ion trap mass spectrometry 446:499 nanospray ionization-linear ion trap mass spectrometry (NSI-MSn) 446:509 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 36 theme ionization-linear 456:472 arg1 NSI-MSn 502:508 NSI-MSn 502:508 NSI-MSn 502:508 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 8 37 theme sialic 1552:1557 arg1 acid 1559:1562 sialic acid 1552:1562 sialic acid 1552:1562 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 0 38 theme Glycans 42:48 arg1 Expression 13:22 Differential Expression 0:22 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 0:82 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 8 39 dep abundant 1463:1470 arg1 NeuAc 1494:1498 NeuAc 1494:1498 NeuAc 1494:1498 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 8 39 dep abundant 1463:1470 arg1 acid 1488:1491 terminal sialic acid 1472:1491 terminal sialic acid (NeuAc) 1472:1499 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 9 40 theme structural 1585:1594 arg1 analyses 1596:1603 Our structural analyses 1581:1603 Our structural analyses of glycans 1581:1614 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 6 41 from structure 994:1002 arg1 % 1055:1055 K100; 57.1% 1045:1055 K100; 57.1% 1045:1055 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 41 from structure 994:1002 arg1 % 1153:1153 M139; 43.0% 1143:1153 M139; 43.0% 1143:1153 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 41 from structure 994:1002 arg1 adenocarcinoma 1029:1042 poorly differentiated adenocarcinoma 1007:1042 poorly differentiated adenocarcinoma (K100; 57.1%) 1007:1056 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 41 from structure 994:1002 arg1 carcinoma 1132:1140 squamous cell carcinoma 1118:1140 squamous cell carcinoma (M139; 43.0%) 1118:1154 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 41 from structure 994:1002 arg1 % 1111:1111 M055; 42.6% 1101:1111 M055; 42.6% 1101:1111 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 41 from structure 994:1002 arg1 adenocarcinoma 1085:1098 moderately differentiated adenocarcinoma 1059:1098 moderately differentiated adenocarcinoma (M055; 42.6%) 1059:1112 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 5 42 theme cell 859:862 arg1 lines 864:868 All five CCA cell lines 846:868 All five CCA cell lines 846:868 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 0 43 gly O-glycoprotein 27:40 arg1 O-glycoprotein 27:40 O-glycoprotein Glycans 27:48 O-glycoprotein Glycans 27:48 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 8 44 link O-linked 1501:1508 arg1 glycans 1510:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 8 45 theme CCA 1443:1445 arg1 lines 1452:1456 All five CCA cell lines 1434:1456 All five CCA cell lines 1434:1456 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 5 46 theme CCA 855:857 arg1 lines 864:868 All five CCA cell lines 846:868 All five CCA cell lines 846:868 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 9 47 theme important 1628:1636 arg1 information 1638:1648 important information 1628:1648 important information regarding physiology of disease-related glycoproteins in CCA 1628:1709 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 8 48 contain have 1458:1461 arg2 glycans 1510:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 8 48 contain have 1458:1461 arg1 lines 1452:1456 All five CCA cell lines 1434:1456 All five CCA cell lines 1434:1456 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 5 49 theme lines 652:656 arg1 glycans 628:634 the O-linked glycans 615:634 the O-linked glycans of the CCA cell lines 615:656 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 3 50 gly glycoprotein 327:338 arg1 glycoprotein 327:338 O-linked glycoprotein 318:338 O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 318:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 6 51 theme cell 1127:1130 arg1 % 1153:1153 M139; 43.0% 1143:1153 M139; 43.0% 1143:1153 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 51 theme cell 1127:1130 arg1 carcinoma 1132:1140 squamous cell carcinoma 1118:1140 squamous cell carcinoma (M139; 43.0%) 1118:1154 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 2 52 theme Aberrant 176:183 arg1 glycosylation 193:205 Aberrant protein glycosylation 176:205 Aberrant protein glycosylation 176:205 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 0 53 theme Cholangiocarcinoma 53:70 arg1 Lines 77:81 Cholangiocarcinoma Cell Lines 53:81 Cholangiocarcinoma Cell Lines 53:81 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 9 54 theme glycoproteins 1690:1702 arg1 physiology 1660:1669 physiology 1660:1669 physiology of disease-related glycoproteins in CCA 1660:1709 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 5 55 theme CCA 643:645 arg1 lines 652:656 the CCA cell lines 639:656 the CCA cell lines 639:656 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 7 56 theme CCA 1429:1431 arg1 types 1420:1424 the different histological types 1393:1424 the different histological types of CCA 1393:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 6 57 dep moderately 1163:1172 arg1 to 1174:1175 to 1174:1175 to 1174:1175 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 5 58 theme glycan 887:892 arg1 pattern 894:900 a similar glycan pattern 877:900 a similar glycan pattern 877:900 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 3 59 theme O-linked 318:325 arg1 glycoprotein 327:338 O-linked glycoprotein 318:338 O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 318:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 9 60 from physiology 1660:1669 arg1 CCA 1707:1709 CCA 1707:1709 CCA 1707:1709 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 8 61 theme abundant 1463:1470 arg1 glycans 1510:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 abundant terminal sialic acid (NeuAc) O-linked glycans 1463:1516 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 6 62 theme adenosquamous 1232:1244 arg1 % 1272:1272 M213; 34.7% 1262:1272 M213; 34.7% 1262:1272 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 62 theme adenosquamous 1232:1244 arg1 carcinoma 1251:1259 adenosquamous cell carcinoma 1232:1259 adenosquamous cell carcinoma (M213; 34.7%) 1232:1273 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 3 63 from types 409:413 arg1 lines 375:379 cholangiocarcinoma (CCA) cell lines 345:379 cholangiocarcinoma (CCA) cell lines from different histological types 345:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 1 64 theme posttranslational 125:141 arg1 glycosylation 92:104 Protein glycosylation 84:104 Protein glycosylation 84:104 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 1 64 theme posttranslational 125:141 arg1 modification 143:154 the most common posttranslational modification 109:154 the most common posttranslational modification in mammalian cells 109:173 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 8 65 theme sialic 1481:1486 arg1 NeuAc 1494:1498 NeuAc 1494:1498 NeuAc 1494:1498 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 8 65 theme sialic 1481:1486 arg1 acid 1488:1491 terminal sialic acid 1472:1491 terminal sialic acid (NeuAc) 1472:1499 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 5 66 theme terminal 698:705 arg1 galactose 707:715 terminal galactose 698:715 terminal galactose 698:715 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 7 67 theme O-linked 1373:1380 arg1 glycans 1382:1388 the O-linked glycans 1369:1388 the O-linked glycans 1369:1388 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 5 68 theme NeuAc2Gal1GalNAc1 774:790 arg1 NeuAc1Gal2GlcNAc1GalNAc1 792:815 NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 774:815 NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 774:815 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 3 69 from lines 375:379 arg1 structures 304:313 the glycan structures 293:313 the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 293:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 69 from lines 375:379 arg1 glycoprotein 327:338 O-linked glycoprotein 318:338 O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types 318:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 5 70 from differences 912:922 arg1 quantities 933:942 their quantities 927:942 their quantities 927:942 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 0 71 from Expression 13:22 arg1 Lines 77:81 Cholangiocarcinoma Cell Lines 53:81 Cholangiocarcinoma Cell Lines 53:81 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 2 72 gly glycosylation 193:205 arg1 diseases 236:243 various diseases 228:243 various diseases 228:243 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 2 72 gly glycosylation 193:205 arg1 cancer 256:261 cancer 256:261 cancer 256:261 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 4 73 theme CCA 523:525 arg1 K100 539:542 K100 539:542 K100 539:542 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 73 theme CCA 523:525 arg1 M055 545:548 M055 545:548 M055 545:548 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 73 theme CCA 523:525 arg1 M214 566:569 M214 566:569 M214 566:569 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 73 theme CCA 523:525 arg1 lines 532:536 Five human CCA cell lines 512:536 Five human CCA cell lines 512:536 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 73 theme CCA 523:525 arg1 M139 551:554 M139 551:554 M139 551:554 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 4 73 theme CCA 523:525 arg1 M213 557:560 M213 557:560 M213 557:560 Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. 26925665 1 74 theme Protein 84:90 arg1 glycosylation 92:104 Protein glycosylation 84:104 Protein glycosylation 84:104 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 1 74 theme Protein 84:90 arg1 modification 143:154 the most common posttranslational modification 109:154 the most common posttranslational modification in mammalian cells 109:173 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 6 75 theme squamous 1118:1125 arg1 % 1153:1153 M139; 43.0% 1143:1153 M139; 43.0% 1143:1153 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 6 75 theme squamous 1118:1125 arg1 carcinoma 1132:1140 squamous cell carcinoma 1118:1140 squamous cell carcinoma (M139; 43.0%) 1118:1154 NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by NeuAc2Gal1GalNAc1. 26925665 3 76 theme different 386:394 arg1 types 409:413 different histological types 386:413 different histological types 386:413 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 8 77 theme important 1533:1541 arg1 role 1543:1546 an important role 1530:1546 an important role for sialic acid in cancer cells 1530:1578 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 5 78 with hexa-saccharides 676:691 arg1 galactose 707:715 terminal galactose 698:715 terminal galactose 698:715 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 5 78 with hexa-saccharides 676:691 arg1 acids 728:732 sialic acids 721:732 sialic acids 721:732 The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. 26925665 3 79 theme mass 483:486 arg1 spectrometry 488:499 nanospray ionization-linear ion trap mass spectrometry 446:499 nanospray ionization-linear ion trap mass spectrometry (NSI-MSn) 446:509 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 79 theme mass 483:486 arg1 NSI-MSn 502:508 NSI-MSn 502:508 NSI-MSn 502:508 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 0 80 theme O-glycoprotein 27:40 arg1 Glycans 42:48 O-glycoprotein Glycans 27:48 O-glycoprotein Glycans 27:48 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 1 81 from modification 143:154 arg1 cells 169:173 mammalian cells 159:173 mammalian cells 159:173 Protein glycosylation is the most common posttranslational modification in mammalian cells. 26925665 7 82 link O-linked 1373:1380 arg1 glycans 1382:1388 the O-linked glycans 1369:1388 the O-linked glycans 1369:1388 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 3 83 theme ion 474:476 arg1 spectrometry 488:499 nanospray ionization-linear ion trap mass spectrometry 446:499 nanospray ionization-linear ion trap mass spectrometry (NSI-MSn) 446:509 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 83 theme ion 474:476 arg1 NSI-MSn 502:508 NSI-MSn 502:508 NSI-MSn 502:508 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 8 84 theme cancer 1567:1572 arg1 cells 1574:1578 cancer cells 1567:1578 cancer cells 1567:1578 All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. 26925665 0 85 theme Cell 72:75 arg1 Lines 77:81 Cholangiocarcinoma Cell Lines 53:81 Cholangiocarcinoma Cell Lines 53:81 Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines. 26925665 3 86 theme nanospray 446:454 arg1 spectrometry 488:499 nanospray ionization-linear ion trap mass spectrometry 446:499 nanospray ionization-linear ion trap mass spectrometry (NSI-MSn) 446:509 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 3 86 theme nanospray 446:454 arg1 NSI-MSn 502:508 NSI-MSn 502:508 NSI-MSn 502:508 We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-MSn). 26925665 9 87 theme glycans 1608:1614 arg1 analyses 1596:1603 Our structural analyses 1581:1603 Our structural analyses of glycans 1581:1614 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26925665 7 88 theme different 1397:1405 arg1 types 1420:1424 the different histological types 1393:1424 the different histological types of CCA 1393:1431 These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. 26925665 2 89 theme various 228:234 arg1 diseases 236:243 various diseases 228:243 various diseases 228:243 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 2 89 theme various 228:234 arg1 cancer 256:261 cancer 256:261 cancer 256:261 Aberrant protein glycosylation has been reported in various diseases, including cancer. 26925665 9 90 gly glycoproteins 1690:1702 arg1 glycoproteins 1690:1702 disease-related glycoproteins 1674:1702 disease-related glycoproteins 1674:1702 Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA. 26483525 10 0 theme relative 1674:1681 arg1 distribution 1683:1694 the relative distribution 1670:1694 the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, 1670:1786 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 5 1 theme branching 685:693 arg1 subsp 714:718 deeply branching Neisseria elongata subsp 678:718 deeply branching Neisseria elongata subsp 678:718 Using a strain of deeply branching Neisseria elongata subsp. 26483525 2 2 theme O-linked 185:192 arg1 glycosylation 202:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 5 3 theme elongata 705:712 arg1 subsp 714:718 deeply branching Neisseria elongata subsp 678:718 deeply branching Neisseria elongata subsp 678:718 Using a strain of deeply branching Neisseria elongata subsp. 26483525 9 4 theme endogenous 1573:1582 arg1 pilus 1592:1596 the endogenous type IV pilus 1569:1596 the endogenous type IV pilus 1569:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 10 5 theme glycosylation 1705:1717 arg1 distribution 1683:1694 the relative distribution 1670:1694 the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, 1670:1786 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 2 6 theme UNLABELLED 159:168 arg1 glycosylation 202:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 17 7 theme evolutionary 2950:2961 arg1 trajectories 2963:2974 the evolutionary trajectories 2946:2974 the evolutionary trajectories 2946:2974 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 7 8 theme glycan 982:987 arg1 serotyping 989:998 glycan serotyping 982:998 glycan serotyping 982:998 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 8 9 theme pglG 1325:1328 arg1 gene 1330:1333 the broadly conserved but enigmatic pglG gene 1289:1333 the broadly conserved but enigmatic pglG gene 1289:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 10 10 theme protein 1741:1747 arg1 substrates 1763:1772 protein glycosylation substrates 1741:1772 protein glycosylation substrates 1741:1772 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 11 11 theme protein 1976:1982 arg1 glycosylation 1984:1996 neisserial O-linked protein glycosylation 1956:1996 neisserial O-linked protein glycosylation diversity 1956:2006 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 17 12 theme PilE 2850:2853 arg1 pilin 2855:2859 PilE pilin 2850:2859 PilE pilin 2850:2859 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 13 13 theme major 2442:2446 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 13 theme major 2442:2446 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 11 14 theme neisserial 1956:1965 arg1 glycosylation 1984:1996 neisserial O-linked protein glycosylation 1956:1996 neisserial O-linked protein glycosylation diversity 1956:2006 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 7 15 theme PglH 1221:1224 arg1 orthologues 1226:1236 glucosyltransferase PglH orthologues 1201:1236 glucosyltransferase PglH orthologues 1201:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 7 16 theme spectrometric 954:966 arg1 analysis 968:975 mass spectrometric analysis 949:975 mass spectrometric analysis 949:975 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 13 17 from components 2234:2243 arg1 systems 2254:2260 these systems 2248:2260 these systems 2248:2260 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 8 18 theme extended 1363:1370 arg1 glycoform 1372:1380 the extended glycoform 1359:1380 the extended glycoform 1359:1380 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 3 19 theme gonorrhoeae 340:350 arg1 strains 319:325 strains 319:325 strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica 319:385 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 2 20 theme Neisseria 251:259 arg1 species 261:267 the major Neisseria species 241:267 the major Neisseria species of importance to human health and disease 241:309 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 7 21 theme successive 1154:1163 arg1 actions 1165:1171 the successive actions 1150:1171 the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1150:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 8 22 theme functional 1434:1443 arg1 product 1421:1427 its product 1417:1427 its product 1417:1427 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 8 22 theme functional 1434:1443 arg1 glycosyltransferase 1445:1463 a functional glycosyltransferase 1432:1463 a functional glycosyltransferase 1432:1463 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 7 23 theme PglC 1182:1185 arg1 actions 1165:1171 the successive actions 1150:1171 the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1150:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 14 24 gly glycosylation 2499:2511 arg1 species 2562:2568 a distantly related, nonpathogenic neisserial species 2516:2568 a distantly related, nonpathogenic neisserial species 2516:2568 Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. 26483525 17 25 theme broad-spectrum 3024:3037 arg1 systems 3061:3067 broad-spectrum protein glycosylation systems 3024:3067 broad-spectrum protein glycosylation systems in bacteria 3024:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 11 26 link O-linked 1967:1974 arg1 glycosylation 1984:1996 neisserial O-linked protein glycosylation 1956:1996 neisserial O-linked protein glycosylation diversity 1956:2006 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 11 27 theme study 1908:1912 arg1 results 1892:1898 the results 1888:1898 the results of this study 1888:1912 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 3 28 theme N. 353:354 arg1 meningitidis 356:367 N. meningitidis 353:367 N. meningitidis 353:367 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 17 29 from trajectories 2963:2974 arg1 bacteria 3072:3079 bacteria 3072:3079 bacteria 3072:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 14 30 theme protein 2491:2497 arg1 glycosylation 2499:2511 protein glycosylation 2491:2511 protein glycosylation 2491:2511 Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. 26483525 0 31 theme O-Linked 79:86 arg1 Glycosylation 96:108 O-Linked Protein Glycosylation 79:108 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 13 32 theme diversifying 2277:2288 arg1 selection 2290:2298 diversifying selection 2277:2298 diversifying selection 2277:2298 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 12 33 theme pathogenic 2114:2123 arg1 gonorrhoeae 2146:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 13 34 theme IV 2464:2465 arg1 pili 2467:2470 type IV pili 2459:2470 type IV pili 2459:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 16 35 theme genus 2779:2783 arg1 level 2785:2789 the genus level 2775:2789 the genus level 2775:2789 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 0 36 from Characterization 0:15 arg1 System 110:115 the O-Linked Protein Glycosylation System 75:115 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 2 37 gly glycosylation 202:214 arg1 species 261:267 the major Neisseria species 241:267 the major Neisseria species of importance to human health and disease 241:309 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 0 38 theme Glycosylation 96:108 arg1 System 110:115 the O-Linked Protein Glycosylation System 75:115 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 12 39 theme species 2135:2141 arg1 gonorrhoeae 2146:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 7 40 theme N. 1102:1103 arg1 gonorrhoeae 1105:1115 N. gonorrhoeae 1102:1115 N. gonorrhoeae 1102:1115 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 3 41 theme N. 374:375 arg1 lactamica 377:385 N. lactamica 374:385 N. lactamica 374:385 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 6 42 theme unrecognized 747:758 arg1 glycolytica 721:731 glycolytica 721:731 glycolytica 721:731 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 6 42 theme unrecognized 747:758 arg1 glycoform 776:784 a heretofore unrecognized tetrasaccharide glycoform 734:784 a heretofore unrecognized tetrasaccharide glycoform 734:784 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 10 43 theme structure-function 1804:1821 arg1 relationships 1823:1835 the overall structure-function relationships 1792:1835 the overall structure-function relationships of broad-spectrum protein glycosylation 1792:1875 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 13 44 theme lines 2191:2195 arg1 number 2181:2186 A number 2179:2186 A number of lines of evidence 2179:2207 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 3 45 theme pgl 411:413 arg1 content 421:427 protein glycosylation (pgl) gene content 388:427 protein glycosylation (pgl) gene content 388:427 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 0 46 from Tetrasaccharide 29:43 arg1 System 110:115 the O-Linked Protein Glycosylation System 75:115 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 3 47 theme corresponding 437:449 arg1 structure 467:475 the corresponding oligosaccharide structure 433:475 the corresponding oligosaccharide structure 433:475 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 13 48 theme PilE 2432:2435 arg1 glycosylation 2366:2378 glycosylation 2366:2378 glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili 2366:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 7 49 theme diNAcBac-Glc-based 1065:1082 arg1 structure 1084:1092 the diNAcBac-Glc-based structure 1061:1092 the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1061:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 3 50 theme glycosylation 396:408 arg1 content 421:427 protein glycosylation (pgl) gene content 388:427 protein glycosylation (pgl) gene content 388:427 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 10 51 theme obvious 1646:1652 arg1 questions 1654:1662 obvious questions 1646:1662 obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, 1646:1786 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 10 52 theme protein 1855:1861 arg1 glycosylation 1863:1875 broad-spectrum protein glycosylation 1840:1875 broad-spectrum protein glycosylation 1840:1875 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 7 53 theme Directed 927:934 arg1 mutagenesis 936:946 Directed mutagenesis 927:946 Directed mutagenesis 927:946 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 12 54 theme protein 2054:2060 arg1 systems 2076:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 16 55 theme novel 2698:2702 arg1 glycoforms 2704:2713 novel glycoforms 2698:2713 novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown 2698:2835 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 17 56 theme overall 2980:2986 arg1 relationships 3007:3019 overall structure-function relationships 2980:3019 overall structure-function relationships 2980:3019 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 4 57 theme related 596:602 arg1 species 614:620 distantly related commensal species 586:620 distantly related commensal species 586:620 The status of such systems in distantly related commensal species, however, remains largely unexplored. 26483525 0 58 theme Tetrasaccharide 29:43 arg1 Characterization 0:15 Characterization 0:15 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 0:144 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 3 59 theme gene 416:419 arg1 content 421:427 protein glycosylation (pgl) gene content 388:427 protein glycosylation (pgl) gene content 388:427 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 2 60 theme protein 194:200 arg1 glycosylation 202:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 0 61 theme Distinct 49:56 arg1 Glycoproteome 58:70 Distinct Glycoproteome 49:70 Distinct Glycoproteome 49:70 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 9 62 theme glycoprotein 1517:1528 arg1 substrates 1530:1539 glycoprotein substrates 1517:1539 glycoprotein substrates 1517:1539 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 4 63 theme such 570:573 arg1 systems 575:581 such systems 570:581 such systems 570:581 The status of such systems in distantly related commensal species, however, remains largely unexplored. 26483525 6 64 theme hexuronic 845:853 arg1 acid-N-acetylhexosamine 855:877 di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine 800:877 di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) 800:909 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 6 64 theme hexuronic 845:853 arg1 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 13 65 theme glycosylation 2366:2378 arg1 context 2355:2361 the context 2351:2361 the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili 2351:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 8 66 theme null 1254:1257 arg1 mutation 1259:1266 a null mutation 1252:1266 a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene 1252:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 16 67 located found 2660:2664 arg1 species 2680:2686 pathogenic species 2669:2686 pathogenic species 2669:2686 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 16 67 located found 2660:2664 arg2 systems 2652:2658 the systems 2648:2658 the systems found in pathogenic species 2648:2686 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 10 68 theme pilin 1699:1703 arg1 glycosylation 1705:1717 pilin glycosylation 1699:1717 pilin glycosylation 1699:1717 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 9 69 theme major 1546:1550 arg1 subunit 1558:1564 the major pilin subunit 1542:1564 the major pilin subunit of the endogenous type IV pilus 1542:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 17 70 from relationships 3007:3019 arg1 bacteria 3072:3079 bacteria 3072:3079 bacteria 3072:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 71 theme glycosylation 3047:3059 arg1 systems 3061:3067 broad-spectrum protein glycosylation systems 3024:3067 broad-spectrum protein glycosylation systems in bacteria 3024:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 16 72 theme clear 2626:2630 arg1 similarities 2632:2643 clear similarities 2626:2643 clear similarities to the systems found in pathogenic species 2626:2686 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 2 73 link O-linked 185:192 arg1 glycosylation 202:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 16 74 contain has 2622:2624 arg2 similarities 2632:2643 clear similarities 2626:2643 clear similarities to the systems found in pathogenic species 2626:2686 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 16 74 contain has 2622:2624 arg1 system 2615:2620 This system 2610:2620 This system 2610:2620 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 14 75 dep related 2528:2534 arg1 nonpathogenic 2537:2549 nonpathogenic 2537:2549 nonpathogenic 2537:2549 Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. 26483525 5 76 theme Neisseria 695:703 arg1 subsp 714:718 deeply branching Neisseria elongata subsp 678:718 deeply branching Neisseria elongata subsp 678:718 Using a strain of deeply branching Neisseria elongata subsp. 26483525 16 77 theme pathogenic 2669:2678 arg1 species 2680:2686 pathogenic species 2669:2686 pathogenic species 2669:2686 This system has clear similarities to the systems found in pathogenic species but makes novel glycoforms utilizing a glycosyltransferase that is widely conserved at the genus level but whose function until now remained unknown. 26483525 2 78 theme Broad-spectrum 170:183 arg1 glycosylation 202:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation 159:214 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 9 79 theme type 1584:1587 arg1 pilus 1592:1596 the endogenous type IV pilus 1569:1596 the endogenous type IV pilus 1569:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 8 80 theme conserved 1301:1309 arg1 gene 1330:1333 the broadly conserved but enigmatic pglG gene 1289:1333 the broadly conserved but enigmatic pglG gene 1289:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 13 81 theme pilin 2418:2422 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 81 theme pilin 2418:2422 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 17 82 theme important 2920:2928 arg1 questions 2930:2938 important questions 2920:2938 important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria 2920:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 8 83 theme glycoform 1372:1380 arg1 expression 1345:1354 expression 1345:1354 expression of the extended glycoform 1345:1380 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 10 84 theme glycosylation 1749:1761 arg1 substrates 1763:1772 protein glycosylation substrates 1741:1772 protein glycosylation substrates 1741:1772 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 17 85 theme systems 3061:3067 arg1 trajectories 2963:2974 the evolutionary trajectories 2946:2974 the evolutionary trajectories 2946:2974 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 85 theme systems 3061:3067 arg1 relationships 3007:3019 overall structure-function relationships 2980:3019 overall structure-function relationships 2980:3019 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 7 86 theme orthologues 1226:1236 arg1 actions 1165:1171 the successive actions 1150:1171 the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1150:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 14 87 theme neisserial 2551:2560 arg1 species 2562:2568 a distantly related, nonpathogenic neisserial species 2516:2568 a distantly related, nonpathogenic neisserial species 2516:2568 Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. 26483525 9 88 theme pilus 1592:1596 arg1 subunit 1558:1564 the major pilin subunit 1542:1564 the major pilin subunit of the endogenous type IV pilus 1542:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 8 89 theme enigmatic 1315:1323 arg1 gene 1330:1333 the broadly conserved but enigmatic pglG gene 1289:1333 the broadly conserved but enigmatic pglG gene 1289:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 11 90 theme glycosylation 1984:1996 arg1 diversity 1998:2006 neisserial O-linked protein glycosylation diversity 1956:2006 neisserial O-linked protein glycosylation diversity 1956:2006 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 0 91 theme subsp 139:143 arg1 System 110:115 the O-Linked Protein Glycosylation System 75:115 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 8 92 theme gene 1330:1333 arg1 orthologue 1275:1284 the orthologue 1271:1284 the orthologue of the broadly conserved but enigmatic pglG gene 1271:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 8 93 theme first 1397:1401 arg1 evidence 1403:1410 the first evidence that its product is a functional glycosyltransferase 1393:1463 the first evidence that its product is a functional glycosyltransferase 1393:1463 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 11 94 theme O-linked 1967:1974 arg1 glycosylation 1984:1996 neisserial O-linked protein glycosylation 1956:1996 neisserial O-linked protein glycosylation diversity 1956:2006 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 7 95 theme PglD 1192:1195 arg1 actions 1165:1171 the successive actions 1150:1171 the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1150:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 2 96 theme importance 272:281 arg1 species 261:267 the major Neisseria species 241:267 the major Neisseria species of importance to human health and disease 241:309 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 13 97 theme protein 2424:2430 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 97 theme protein 2424:2430 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 11 98 theme genus 2015:2019 arg1 level 2021:2025 the genus level 2011:2025 the genus level 2011:2025 Together, the results of this study provide a foundation upon which to assess neisserial O-linked protein glycosylation diversity at the genus level. 26483525 5 99 theme subsp 714:718 arg1 strain 668:673 a strain 666:673 a strain of deeply branching Neisseria elongata subsp 666:718 Using a strain of deeply branching Neisseria elongata subsp. 26483525 7 100 theme PglB 1176:1179 arg1 actions 1165:1171 the successive actions 1150:1171 the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1150:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 13 101 theme pili 2467:2470 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 101 theme pili 2467:2470 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 12 102 theme N. 2162:2163 arg1 meningitidis 2165:2176 N. meningitidis 2162:2176 N. meningitidis 2162:2176 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 7 103 theme glucosyltransferase 1201:1219 arg1 orthologues 1226:1236 glucosyltransferase PglH orthologues 1201:1236 glucosyltransferase PglH orthologues 1201:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 9 104 gly glycosylated 1606:1617 arg1 subunit 1558:1564 the major pilin subunit 1542:1564 the major pilin subunit of the endogenous type IV pilus 1542:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 14 105 theme related 2528:2534 arg1 species 2562:2568 a distantly related, nonpathogenic neisserial species 2516:2568 a distantly related, nonpathogenic neisserial species 2516:2568 Here, we examined protein glycosylation in a distantly related, nonpathogenic neisserial species, Neisseria elongata subsp. 26483525 3 106 theme Neisseria 330:338 arg1 gonorrhoeae 340:350 Neisseria gonorrhoeae 330:350 Neisseria gonorrhoeae 330:350 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 0 107 from System 110:115 arg1 Characterization 0:15 Characterization 0:15 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 0:144 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 13 108 theme glycan 2317:2322 arg1 variation 2324:2332 glycan variation 2317:2332 glycan variation 2317:2332 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 2 109 theme human 286:290 arg1 health 292:297 human health 286:297 human health 286:297 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 6 110 gly glycoform 776:784 arg1 tetrasaccharide 760:774 a heretofore unrecognized tetrasaccharide glycoform 734:784 a heretofore unrecognized tetrasaccharide glycoform 734:784 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 17 111 theme protein 3039:3045 arg1 systems 3061:3067 broad-spectrum protein glycosylation systems 3024:3067 broad-spectrum protein glycosylation systems in bacteria 3024:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 4 112 from status 560:565 arg1 species 614:620 distantly related commensal species 586:620 distantly related commensal species 586:620 The status of such systems in distantly related commensal species, however, remains largely unexplored. 26483525 17 113 gly glycosylated 2868:2879 arg1 finding 2900:2906 a finding 2898:2906 a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria 2898:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 113 gly glycosylated 2868:2879 arg1 pilin 2855:2859 PilE pilin 2850:2859 PilE pilin 2850:2859 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 113 gly glycosylated 2868:2879 arg2 pilin 2855:2859 PilE pilin 2850:2859 PilE pilin 2850:2859 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 113 gly glycosylated 2868:2879 arg1 species 2889:2895 this species 2884:2895 this species 2884:2895 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 0 114 theme Protein 88:94 arg1 Glycosylation 96:108 O-Linked Protein Glycosylation 79:108 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 12 115 theme Neisseria 2125:2133 arg1 gonorrhoeae 2146:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 12 116 theme IMPORTANCE 2028:2037 arg1 systems 2076:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 13 117 theme type 2459:2462 arg1 pili 2467:2470 type IV pili 2459:2470 type IV pili 2459:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 8 118 from mutation 1259:1266 arg1 orthologue 1275:1284 the orthologue 1271:1284 the orthologue of the broadly conserved but enigmatic pglG gene 1271:1333 In addition, a null mutation in the orthologue of the broadly conserved but enigmatic pglG gene precluded expression of the extended glycoform, providing the first evidence that its product is a functional glycosyltransferase. 26483525 3 119 theme meningitidis 356:367 arg1 strains 319:325 strains 319:325 strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica 319:385 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 12 120 theme N. 2143:2144 arg1 gonorrhoeae 2146:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 the pathogenic Neisseria species N. gonorrhoeae 2110:2156 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 7 121 theme N. 1121:1122 arg1 meningitidis 1124:1135 N. meningitidis 1121:1135 N. meningitidis 1121:1135 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 3 122 theme protein 388:394 arg1 content 421:427 protein glycosylation (pgl) gene content 388:427 protein glycosylation (pgl) gene content 388:427 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 0 123 from Glycoproteome 58:70 arg1 System 110:115 the O-Linked Protein Glycosylation System 75:115 the O-Linked Protein Glycosylation System of Neisseria elongata subsp 75:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 6 124 theme tetrasaccharide 760:774 arg1 glycolytica 721:731 glycolytica 721:731 glycolytica 721:731 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 6 124 theme tetrasaccharide 760:774 arg1 glycoform 776:784 a heretofore unrecognized tetrasaccharide glycoform 734:784 a heretofore unrecognized tetrasaccharide glycoform 734:784 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 0 125 theme Neisseria 120:128 arg1 subsp 139:143 Neisseria elongata subsp 120:143 Neisseria elongata subsp 120:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 3 126 theme lactamica 377:385 arg1 strains 319:325 strains 319:325 strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica 319:385 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 10 127 theme overall 1796:1802 arg1 relationships 1823:1835 the overall structure-function relationships 1792:1835 the overall structure-function relationships of broad-spectrum protein glycosylation 1792:1875 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 9 128 theme clear 1474:1478 arg1 evidence 1480:1487 clear evidence 1474:1487 clear evidence for a substantial number of glycoprotein substrates 1474:1539 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 10 129 dep genus 1730:1734 arg1 selected 1778:1785 selected 1778:1785 are selected 1774:1785 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 3 130 theme oligosaccharide 451:465 arg1 structure 467:475 the corresponding oligosaccharide structure 433:475 the corresponding oligosaccharide structure 433:475 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 0 131 theme Unique 22:27 arg1 Tetrasaccharide 29:43 a Unique Tetrasaccharide 20:43 a Unique Tetrasaccharide 20:43 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 7 132 theme mass 949:952 arg1 analysis 968:975 mass spectrometric analysis 949:975 mass spectrometric analysis 949:975 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 12 133 theme glycosylation 2062:2074 arg1 systems 2076:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 7 134 theme structure 1084:1092 arg1 version 1050:1056 an extended version 1038:1056 an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1038:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 7 134 theme structure 1084:1092 arg1 oligosaccharide 1019:1033 the oligosaccharide 1015:1033 the oligosaccharide 1015:1033 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 4 135 theme commensal 604:612 arg1 species 614:620 distantly related commensal species 586:620 distantly related commensal species 586:620 The status of such systems in distantly related commensal species, however, remains largely unexplored. 26483525 9 136 gly glycoprotein 1517:1528 arg1 glycoprotein 1517:1528 glycoprotein substrates 1517:1539 glycoprotein substrates 1517:1539 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 10 137 theme broad-spectrum 1840:1853 arg1 glycosylation 1863:1875 broad-spectrum protein glycosylation 1840:1875 broad-spectrum protein glycosylation 1840:1875 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 9 138 theme substantial 1495:1505 arg1 number 1507:1512 a substantial number 1493:1512 a substantial number of glycoprotein substrates 1493:1539 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 17 139 theme structure-function 2988:3005 arg1 relationships 3007:3019 overall structure-function relationships 2980:3019 overall structure-function relationships 2980:3019 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 10 140 theme latter 1624:1629 arg1 finding 1631:1637 The latter finding 1620:1637 The latter finding 1620:1637 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 13 141 gly glycosylation 2366:2378 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 141 gly glycosylation 2366:2378 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 0 142 theme Glycoproteome 58:70 arg1 Characterization 0:15 Characterization 0:15 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 0:144 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 13 143 theme surface-localized 2400:2416 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 143 theme surface-localized 2400:2416 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 17 144 from bacteria 3072:3079 arg1 trajectories 2963:2974 the evolutionary trajectories 2946:2974 the evolutionary trajectories 2946:2974 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 17 144 from bacteria 3072:3079 arg1 relationships 3007:3019 overall structure-function relationships 2980:3019 overall structure-function relationships 2980:3019 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 4 145 theme systems 575:581 arg1 status 560:565 The status 556:565 The status of such systems in distantly related commensal species 556:620 The status of such systems in distantly related commensal species, however, remains largely unexplored. 26483525 2 146 theme major 245:249 arg1 species 261:267 the major Neisseria species 241:267 the major Neisseria species of importance to human health and disease 241:309 UNLABELLED Broad-spectrum O-linked protein glycosylation is well characterized in the major Neisseria species of importance to human health and disease. 26483525 13 147 theme glycan 2227:2232 arg1 subject 2266:2272 subject 2266:2272 subject 2266:2272 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 147 theme glycan 2227:2232 arg1 components 2234:2243 the glycan components 2223:2243 the glycan components in these systems 2223:2260 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 3 148 theme interstrain 524:534 arg1 variability 536:546 interstrain variability 524:546 interstrain variability 524:546 Within strains of Neisseria gonorrhoeae, N. meningitidis, and N. lactamica, protein glycosylation (pgl) gene content and the corresponding oligosaccharide structure are fairly well conserved, although intra- and interstrain variability occurs. 26483525 9 149 theme substrates 1530:1539 arg1 number 1507:1512 a substantial number 1493:1512 a substantial number of glycoprotein substrates 1493:1539 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26483525 12 150 theme Broad-spectrum 2039:2052 arg1 systems 2076:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems 2028:2082 IMPORTANCE Broad-spectrum protein glycosylation systems are well characterized in the pathogenic Neisseria species N. gonorrhoeae and N. meningitidis. 26483525 17 151 from systems 3061:3067 arg1 bacteria 3072:3079 bacteria 3072:3079 bacteria 3072:3079 Remarkably, PilE pilin is not glycosylated in this species, a finding that raises important questions about the evolutionary trajectories and overall structure-function relationships of broad-spectrum protein glycosylation systems in bacteria. 26483525 6 152 theme di-N-acetylbacillosamine-glucose-di-N-acetyl 800:843 arg1 acid-N-acetylhexosamine 855:877 di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine 800:877 di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) 800:909 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 6 152 theme di-N-acetylbacillosamine-glucose-di-N-acetyl 800:843 arg1 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 diNAcBac-Glc-diNAcHexA-HexNAc 880:908 glycolytica, a heretofore unrecognized tetrasaccharide glycoform consisting of di-N-acetylbacillosamine-glucose-di-N-acetyl hexuronic acid-N-acetylhexosamine (diNAcBac-Glc-diNAcHexA-HexNAc) was identified. 26483525 10 153 theme glycosylation 1863:1875 arg1 relationships 1823:1835 the overall structure-function relationships 1792:1835 the overall structure-function relationships of broad-spectrum protein glycosylation 1792:1875 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 10 153 theme glycosylation 1863:1875 arg1 questions 1654:1662 obvious questions 1646:1662 obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, 1646:1786 The latter finding raises obvious questions as to the relative distribution of pilin glycosylation within the genus, how protein glycosylation substrates are selected, and the overall structure-function relationships of broad-spectrum protein glycosylation. 26483525 7 154 theme extended 1041:1048 arg1 version 1050:1056 an extended version 1038:1056 an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues 1038:1236 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 7 154 theme extended 1041:1048 arg1 oligosaccharide 1019:1033 the oligosaccharide 1015:1033 the oligosaccharide 1015:1033 Directed mutagenesis, mass spectrometric analysis, and glycan serotyping confirmed that the oligosaccharide is an extended version of the diNAcBac-Glc-based structure seen in N. gonorrhoeae and N. meningitidis generated by the successive actions of PglB, PglC, and PglD and glucosyltransferase PglH orthologues. 26483525 0 155 theme elongata 130:137 arg1 subsp 139:143 Neisseria elongata subsp 120:143 Neisseria elongata subsp 120:143 Characterization of a Unique Tetrasaccharide and Distinct Glycoproteome in the O-Linked Protein Glycosylation System of Neisseria elongata subsp. 26483525 13 156 theme abundant 2387:2394 arg1 subunit 2448:2454 the major subunit 2438:2454 the major subunit of type IV pili 2438:2470 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 156 theme abundant 2387:2394 arg1 PilE 2432:2435 the abundant and surface-localized pilin protein PilE 2383:2435 the abundant and surface-localized pilin protein PilE 2383:2435 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 13 157 theme evidence 2200:2207 arg1 lines 2191:2195 lines 2191:2195 lines of evidence 2191:2207 A number of lines of evidence indicate that the glycan components in these systems are subject to diversifying selection and suggest that glycan variation may be driven in the context of glycosylation of the abundant and surface-localized pilin protein PilE, the major subunit of type IV pili. 26483525 9 158 theme pilin 1552:1556 arg1 subunit 1558:1564 the major pilin subunit 1542:1564 the major pilin subunit of the endogenous type IV pilus 1542:1596 Despite clear evidence for a substantial number of glycoprotein substrates, the major pilin subunit of the endogenous type IV pilus was not glycosylated. 26077366 0 0 theme molecular 128:136 arg1 simulations 147:157 thermal perturbation molecular dynamics simulations 107:157 thermal perturbation molecular dynamics simulations 107:157 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 1 from simulations 147:157 arg1 insight 94:100 an insight 91:100 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 0:158 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 10 2 theme peptide 1533:1539 arg1 orientation 1547:1557 corresponding cis peptide angle orientation 1515:1557 corresponding cis peptide angle orientation 1515:1557 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 2 3 theme conformational 349:362 arg1 transitions 364:374 the protein conformational transitions 337:374 the protein conformational transitions 337:374 Its effect on the protein conformational transitions is not clearly understood yet. 26077366 1 4 theme post-translational 235:252 arg1 modifications 254:266 the most prevalent and complex post-translational modifications 204:266 the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates 204:320 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 2 5 from effect 327:332 arg1 transitions 364:374 the protein conformational transitions 337:374 the protein conformational transitions 337:374 Its effect on the protein conformational transitions is not clearly understood yet. 26077366 7 6 theme sheets 1118:1123 arg1 loops 1082:1086 solvent exposed loops 1066:1086 solvent exposed loops 1066:1086 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 7 6 theme sheets 1118:1123 arg1 ends 1099:1102 ends 1099:1102 ends 1099:1102 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 7 6 theme sheets 1118:1123 arg1 turns 1089:1093 turns 1089:1093 turns 1089:1093 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 6 7 gly glycosylated 932:943 arg1 form 945:948 corresponding glycosylated form 918:948 corresponding glycosylated form at normal and elevated temperatures 918:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 8 8 theme important 1172:1180 arg1 saltbridges 1182:1192 some important saltbridges 1167:1192 some important saltbridges 1167:1192 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 8 8 theme important 1172:1180 arg1 bonds 1204:1208 hydrogen bonds 1195:1208 hydrogen bonds 1195:1208 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 8 8 theme important 1172:1180 arg1 interactions 1226:1237 hydrophobic interactions 1214:1237 hydrophobic interactions within the protein 1214:1256 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 7 9 theme distinct 1107:1114 arg1 sheets 1118:1123 distinct β sheets 1107:1123 distinct β sheets 1107:1123 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 3 10 theme soybean 541:547 arg1 agglutinin 549:558 legume lectin soybean agglutinin 527:558 legume lectin soybean agglutinin (SBA) 527:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 3 10 theme soybean 541:547 arg1 SBA 561:563 SBA 561:563 SBA 561:563 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 6 11 theme nonglycosylated 867:881 arg1 form 893:896 the nonglycosylated monomeric form 863:896 the nonglycosylated monomeric form 863:896 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 6 11 theme nonglycosylated 867:881 arg1 stable 906:911 stable 906:911 stable 906:911 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 0 12 theme perturbation 115:126 arg1 simulations 147:157 thermal perturbation molecular dynamics simulations 107:157 thermal perturbation molecular dynamics simulations 107:157 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 7 13 from temperature 1009:1019 arg1 prominent 1053:1061 prominent 1053:1061 prominent 1053:1061 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 8 14 theme hydrophobic 1214:1224 arg1 saltbridges 1182:1192 some important saltbridges 1167:1192 some important saltbridges 1167:1192 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 8 14 theme hydrophobic 1214:1224 arg1 interactions 1226:1237 hydrophobic interactions 1214:1237 hydrophobic interactions within the protein 1214:1256 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 7 15 from loops 1082:1086 arg1 prominent 1053:1061 prominent 1053:1061 prominent 1053:1061 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 11 16 contain have 1593:1596 arg1 description 1575:1585 This atomistic description 1560:1585 This atomistic description 1560:1585 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 11 16 contain have 1593:1596 arg2 implications 1608:1619 important implications 1598:1619 important implications for understanding the functionality and stability of Soybean agglutinin 1598:1691 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 7 17 theme solvent 1066:1072 arg1 loops 1082:1086 solvent exposed loops 1066:1086 solvent exposed loops 1066:1086 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 9 18 theme reducing 1263:1270 arg1 residues 1288:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 10 19 contain has 1486:1488 arg1 glycosylation 1464:1476 single glycosylation 1457:1476 single glycosylation (ASN75) 1457:1484 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 10 19 contain has 1486:1488 arg2 effect 1505:1510 no significant effect 1490:1510 no significant effect 1490:1510 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 10 19 contain has 1486:1488 arg1 ASN75 1479:1483 ASN75 1479:1483 ASN75 1479:1483 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 6 20 theme glycosylated 932:943 arg1 form 945:948 corresponding glycosylated form 918:948 corresponding glycosylated form at normal and elevated temperatures 918:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 5 21 theme thermal 698:704 arg1 unfolding 706:714 thermal unfolding 698:714 thermal unfolding of these two forms 698:733 To unveil the structural perturbations during thermal unfolding of these two forms, we have studied and compared them to the experimental results. 26077366 9 22 theme GlcNAc 1281:1286 arg1 residues 1288:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 0 23 theme dynamics 138:145 arg1 simulations 147:157 thermal perturbation molecular dynamics simulations 107:157 thermal perturbation molecular dynamics simulations 107:157 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 24 theme soybean 65:71 arg1 SBA 85:87 SBA 85:87 SBA 85:87 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 24 theme soybean 65:71 arg1 agglutinin 73:82 soybean agglutinin 65:82 soybean agglutinin (SBA) 65:88 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 25 from Impact 0:5 arg1 unfolding 52:60 unfolding 52:60 unfolding 52:60 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 25 from Impact 0:5 arg1 structure 38:46 structure 38:46 structure 38:46 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 25 from Impact 0:5 arg1 stability 27:35 stability 27:35 stability 27:35 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 4 26 gly glycosylated 571:582 arg1 moiety 584:589 Its glycosylated moiety 567:589 Its glycosylated moiety 567:589 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 1 27 theme proteins 271:278 arg1 modifications 254:266 the most prevalent and complex post-translational modifications 204:266 the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates 204:320 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 7 28 from ends 1099:1102 arg1 prominent 1053:1061 prominent 1053:1061 prominent 1053:1061 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 10 29 theme significant 1493:1503 arg1 effect 1505:1510 no significant effect 1490:1510 no significant effect 1490:1510 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 3 30 theme glycosylation 453:465 arg1 effect 443:448 the effect 439:448 the effect of glycosylation on protein stability 439:486 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 0 31 theme glycosylation 10:22 arg1 Impact 0:5 Impact 0:5 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 0:158 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 1 32 theme numerous 290:297 arg1 enzymes 299:305 numerous enzymes 290:305 numerous enzymes 290:305 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 4 33 link N-linked 621:628 arg1 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 33 link N-linked 621:628 arg1 glycan 630:635 high mannose type N-linked glycan 603:635 high mannose type N-linked glycan (Man9GlcNAc2) 603:649 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 10 34 theme cis 1529:1531 arg1 orientation 1547:1557 corresponding cis peptide angle orientation 1515:1557 corresponding cis peptide angle orientation 1515:1557 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 2 35 theme protein 341:347 arg1 transitions 364:374 the protein conformational transitions 337:374 the protein conformational transitions 337:374 Its effect on the protein conformational transitions is not clearly understood yet. 26077366 9 36 theme hydrogen 1382:1389 arg1 bonding 1391:1397 hydrogen bonding 1382:1397 hydrogen bonding with ASN 9 and ASN 75 1382:1419 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 11 37 theme important 1598:1606 arg1 implications 1608:1619 important implications 1598:1619 important implications for understanding the functionality and stability of Soybean agglutinin 1598:1691 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 3 38 theme protein 470:476 arg1 stability 478:486 protein stability 470:486 protein stability 470:486 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 6 39 theme dynamics 823:830 arg1 perspective 808:818 the perspective 804:818 the perspective of dynamics 804:830 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 7 40 from prominent 1053:1061 arg1 loops 1082:1086 solvent exposed loops 1066:1086 solvent exposed loops 1066:1086 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 7 40 from prominent 1053:1061 arg1 ends 1099:1102 ends 1099:1102 ends 1099:1102 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 7 40 from prominent 1053:1061 arg1 turns 1089:1093 turns 1089:1093 turns 1089:1093 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 1 41 theme prevalent 213:221 arg1 modifications 254:266 the most prevalent and complex post-translational modifications 204:266 the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates 204:320 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 6 42 theme corresponding 918:930 arg1 form 945:948 corresponding glycosylated form 918:948 corresponding glycosylated form at normal and elevated temperatures 918:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 0 43 dep Impact 0:5 arg1 insight 94:100 an insight 91:100 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 0:158 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 4 44 theme type 616:619 arg1 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 44 theme type 616:619 arg1 glycan 630:635 high mannose type N-linked glycan 603:635 high mannose type N-linked glycan (Man9GlcNAc2) 603:649 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 10 45 theme angle 1541:1545 arg1 orientation 1547:1557 corresponding cis peptide angle orientation 1515:1557 corresponding cis peptide angle orientation 1515:1557 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 11 46 theme agglutinin 1682:1691 arg1 stability 1661:1669 stability 1661:1669 stability 1661:1669 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 11 46 theme agglutinin 1682:1691 arg1 functionality 1643:1655 functionality 1643:1655 functionality 1643:1655 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 3 47 theme dynamics 504:511 arg1 simulation 513:522 molecular dynamics simulation 494:522 molecular dynamics simulation of legume lectin soybean agglutinin (SBA) 494:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 1 48 theme complex 227:233 arg1 modifications 254:266 the most prevalent and complex post-translational modifications 204:266 the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates 204:320 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 8 49 theme folded 1143:1148 arg1 structure 1150:1158 it folded structure 1140:1158 it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein 1140:1256 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 4 50 theme glycosylated 571:582 arg1 moiety 584:589 Its glycosylated moiety 567:589 Its glycosylated moiety 567:589 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 7 51 theme β 1116:1116 arg1 sheets 1118:1123 distinct β sheets 1107:1123 distinct β sheets 1107:1123 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 9 52 with bonding 1391:1397 arg1 ASN 1404:1406 ASN 9 1404:1408 ASN 9 1404:1408 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 52 with bonding 1391:1397 arg1 ASN 1414:1416 ASN 75 1414:1419 ASN 75 1414:1419 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 3 53 theme agglutinin 549:558 arg1 simulation 513:522 molecular dynamics simulation 494:522 molecular dynamics simulation of legume lectin soybean agglutinin (SBA) 494:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 1 54 theme modifications 254:266 arg1 modifications 254:266 the most prevalent and complex post-translational modifications 204:266 the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates 204:320 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 1 54 theme modifications 254:266 arg1 one 197:199 one 197:199 one 197:199 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 1 54 theme modifications 254:266 arg1 Glycosylation 160:172 Glycosylation 160:172 Glycosylation 160:172 Glycosylation has been recognized as one of the most prevalent and complex post-translational modifications of proteins involving numerous enzymes and substrates. 26077366 8 55 theme due 1160:1162 arg1 structure 1150:1158 it folded structure 1140:1158 it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein 1140:1256 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 7 56 from turns 1089:1093 arg1 prominent 1053:1061 prominent 1053:1061 prominent 1053:1061 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 3 57 theme lectin 534:539 arg1 agglutinin 549:558 legume lectin soybean agglutinin 527:558 legume lectin soybean agglutinin (SBA) 527:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 3 57 theme lectin 534:539 arg1 SBA 561:563 SBA 561:563 SBA 561:563 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 6 58 theme monomeric 883:891 arg1 form 893:896 the nonglycosylated monomeric form 863:896 the nonglycosylated monomeric form 863:896 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 6 58 theme monomeric 883:891 arg1 stable 906:911 stable 906:911 stable 906:911 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 11 59 theme atomistic 1565:1573 arg1 description 1575:1585 This atomistic description 1560:1585 This atomistic description 1560:1585 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 8 60 theme hydrogen 1195:1202 arg1 saltbridges 1182:1192 some important saltbridges 1167:1192 some important saltbridges 1167:1192 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 8 60 theme hydrogen 1195:1202 arg1 bonds 1204:1208 hydrogen bonds 1195:1208 hydrogen bonds 1195:1208 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 6 61 theme elevated 964:971 arg1 temperatures 973:984 normal and elevated temperatures 953:984 normal and elevated temperatures 953:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 11 62 dep functionality 1643:1655 arg1 the 1639:1641 the 1639:1641 the 1639:1641 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 7 63 theme exposed 1074:1080 arg1 loops 1082:1086 solvent exposed loops 1066:1086 solvent exposed loops 1066:1086 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 6 64 theme normal 953:958 arg1 temperatures 973:984 normal and elevated temperatures 953:984 normal and elevated temperatures 953:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 5 65 theme structural 666:675 arg1 perturbations 677:689 the structural perturbations 662:689 the structural perturbations during thermal unfolding of these two forms 662:733 To unveil the structural perturbations during thermal unfolding of these two forms, we have studied and compared them to the experimental results. 26077366 9 66 theme terminal 1272:1279 arg1 residues 1288:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues 1259:1295 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 7 67 theme thermal 1021:1027 arg1 destabilization 1029:1043 thermal destabilization 1021:1043 thermal destabilization 1021:1043 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 6 68 gly nonglycosylated 867:881 arg1 form 893:896 the nonglycosylated monomeric form 863:896 the nonglycosylated monomeric form 863:896 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 6 68 gly nonglycosylated 867:881 arg1 stable 906:911 stable 906:911 stable 906:911 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 7 69 theme elevated 1000:1007 arg1 temperature 1009:1019 elevated temperature 1000:1019 elevated temperature 1000:1019 Moreover, at elevated temperature thermal destabilization is more prominent in solvent exposed loops, turns and ends of distinct β sheets. 26077366 0 70 theme agglutinin 73:82 arg1 unfolding 52:60 unfolding 52:60 unfolding 52:60 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 70 theme agglutinin 73:82 arg1 structure 38:46 structure 38:46 structure 38:46 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 0 70 theme agglutinin 73:82 arg1 stability 27:35 stability 27:35 stability 27:35 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 5 71 theme forms 729:733 arg1 unfolding 706:714 thermal unfolding 698:714 thermal unfolding of these two forms 698:733 To unveil the structural perturbations during thermal unfolding of these two forms, we have studied and compared them to the experimental results. 26077366 9 72 theme protein 1315:1321 arg1 VAL161 1332:1337 VAL161 1332:1337 VAL161 1332:1337 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 72 theme protein 1315:1321 arg1 SER225 1351:1356 SER225 1351:1356 SER225 1351:1356 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 72 theme protein 1315:1321 arg1 residues 1323:1330 the protein residues VAL161, PRO182 and SER225 1311:1356 the protein residues VAL161, PRO182 and SER225 1311:1356 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 72 theme protein 1315:1321 arg1 PRO182 1340:1345 PRO182 1340:1345 PRO182 1340:1345 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 8 73 dep folded 1143:1148 arg1 it 1140:1141 it 1140:1141 it 1140:1141 SBA maintains it folded structure due to some important saltbridges, hydrogen bonds and hydrophobic interactions within the protein. 26077366 4 74 theme high 603:606 arg1 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 74 theme high 603:606 arg1 glycan 630:635 high mannose type N-linked glycan 603:635 high mannose type N-linked glycan (Man9GlcNAc2) 603:649 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 0 75 theme thermal 107:113 arg1 simulations 147:157 thermal perturbation molecular dynamics simulations 107:157 thermal perturbation molecular dynamics simulations 107:157 Impact of glycosylation on stability, structure and unfolding of soybean agglutinin (SBA): an insight from thermal perturbation molecular dynamics simulations. 26077366 10 76 theme single 1457:1462 arg1 glycosylation 1464:1476 single glycosylation 1457:1476 single glycosylation (ASN75) 1457:1484 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 10 76 theme single 1457:1462 arg1 ASN75 1479:1483 ASN75 1479:1483 ASN75 1479:1483 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 6 77 from temperatures 973:984 arg1 form 945:948 corresponding glycosylated form 918:948 corresponding glycosylated form at normal and elevated temperatures 918:984 From the perspective of dynamics, our simulations revealed that the nonglycosylated monomeric form is less stable than corresponding glycosylated form at normal and elevated temperatures. 26077366 3 78 gly glycosylation 453:465 arg1 protein 470:476 protein stability 470:486 protein stability 470:486 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 3 79 from effect 443:448 arg1 stability 478:486 protein stability 470:486 protein stability 470:486 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 5 80 theme experimental 777:788 arg1 results 790:796 the experimental results 773:796 the experimental results 773:796 To unveil the structural perturbations during thermal unfolding of these two forms, we have studied and compared them to the experimental results. 26077366 9 81 dep residues 1323:1330 arg1 VAL161 1332:1337 VAL161 1332:1337 VAL161 1332:1337 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 81 dep residues 1323:1330 arg1 SER225 1351:1356 SER225 1351:1356 SER225 1351:1356 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 81 dep residues 1323:1330 arg1 residues 1323:1330 the protein residues VAL161, PRO182 and SER225 1311:1356 the protein residues VAL161, PRO182 and SER225 1311:1356 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 9 81 dep residues 1323:1330 arg1 PRO182 1340:1345 PRO182 1340:1345 PRO182 1340:1345 The reducing terminal GlcNAc residues interact with the protein residues VAL161, PRO182 and SER225 via hydrophobic and via hydrogen bonding with ASN 9 and ASN 75. 26077366 10 82 theme corresponding 1515:1527 arg1 orientation 1547:1557 corresponding cis peptide angle orientation 1515:1557 corresponding cis peptide angle orientation 1515:1557 Our simulations also revealed that single glycosylation (ASN75) has no significant effect on corresponding cis peptide angle orientation. 26077366 3 83 theme molecular 494:502 arg1 simulation 513:522 molecular dynamics simulation 494:522 molecular dynamics simulation of legume lectin soybean agglutinin (SBA) 494:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 4 84 theme mannose 608:614 arg1 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 84 theme mannose 608:614 arg1 glycan 630:635 high mannose type N-linked glycan 603:635 high mannose type N-linked glycan (Man9GlcNAc2) 603:649 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 85 theme N-linked 621:628 arg1 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Man9GlcNAc2 638:648 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 4 85 theme N-linked 621:628 arg1 glycan 630:635 high mannose type N-linked glycan 603:635 high mannose type N-linked glycan (Man9GlcNAc2) 603:649 Its glycosylated moiety consists of high mannose type N-linked glycan (Man9GlcNAc2). 26077366 11 86 theme Soybean 1674:1680 arg1 agglutinin 1682:1691 Soybean agglutinin 1674:1691 Soybean agglutinin 1674:1691 This atomistic description might have important implications for understanding the functionality and stability of Soybean agglutinin. 26077366 3 87 theme legume 527:532 arg1 agglutinin 549:558 legume lectin soybean agglutinin 527:558 legume lectin soybean agglutinin (SBA) 527:564 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 26077366 3 87 theme legume 527:532 arg1 SBA 561:563 SBA 561:563 SBA 561:563 In this study, we have examined the effect of glycosylation on protein stability using molecular dynamics simulation of legume lectin soybean agglutinin (SBA). 28576848 0 0 theme Receptor 27:34 arg1 Glycoforms 36:45 Fc Gamma Receptor Glycoforms 18:45 Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab 18:102 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 7 1 from source 1695:1700 arg1 FcγRI 1679:1683 FcγRI 1679:1683 FcγRI from each source 1679:1700 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 3 2 theme hamster 628:634 arg1 CHO 643:645 CHO 643:645 CHO 643:645 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 2 theme hamster 628:634 arg1 ovary 636:640 Chinese hamster ovary 620:640 Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 620:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 4 3 theme CHO-FcγRIIIaPhe158/Val158 938:962 arg1 interactions 978:989 the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions 934:989 the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions 934:989 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 3 4 theme human 658:662 arg1 receptors 668:676 human Fcγ receptors 658:676 human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 658:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 4 theme human 658:662 arg1 RIIb 716:719 RIIb 716:719 RIIb (CD32b) 716:727 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 4 theme human 658:662 arg1 RIIaArg131/His131 689:705 RIIaArg131/His131 689:705 RIIaArg131/His131 (CD32a) 689:713 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 4 theme human 658:662 arg1 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 (CD16a) 733:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 4 theme human 658:662 arg1 RI 678:679 RI (CD64) 678:686 RI (CD64) 678:686 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 4 5 dep murine 1063:1068 arg1 NS0 1071:1073 NS0 1071:1073 NS0 1071:1073 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 9 6 theme FcγR 2056:2059 arg1 glycosylation 2035:2047 the glycosylation 2031:2047 the glycosylation of the FcγR 2031:2059 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 1 7 theme antibodies 153:162 arg1 region 143:148 the Fc region 136:148 the Fc region of antibodies 136:162 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 8 8 gly sialylated 1831:1840 arg1 glycans 1842:1848 large and sialylated glycans 1821:1848 large and sialylated glycans 1821:1848 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 2 9 theme immune 282:287 arg1 cell 298:301 The immune effector cell 278:301 The immune effector cell activity 278:310 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 6 10 theme HEK293 1411:1416 arg1 cells 1418:1422 NS0 and HEK293 cells 1403:1422 cells 1418:1422 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 6 11 theme NS0 1403:1405 arg1 cells 1418:1422 NS0 and HEK293 cells 1403:1422 cells 1418:1422 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 3 12 with mechanism 811:819 arg1 IgG 826:828 IgG 826:828 IgG 826:828 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 6 13 theme FcγRIIIaPhe158/Val158 1312:1332 arg1 profiles 1276:1283 The glycan profiles 1265:1283 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 1265:1332 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 2 14 theme FcγR 523:526 arg1 glycans 528:534 the FcγR glycans 519:534 the FcγR glycans 519:534 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 9 15 theme FcγR 2103:2106 arg1 glycans 2108:2114 FcγR glycans 2103:2114 FcγR glycans 2103:2114 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 7 16 theme Oligomannose 1638:1649 arg1 structures 1651:1660 Oligomannose structures 1638:1660 Oligomannose structures 1638:1660 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 5 17 from differences 1210:1220 arg1 glycosylation 1250:1262 host cell-dependent FcγR glycosylation 1225:1262 host cell-dependent FcγR glycosylation 1225:1262 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 8 18 contain have 1850:1853 arg2 impact 1866:1871 a negative impact 1855:1871 a negative impact 1855:1871 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 8 18 contain have 1850:1853 arg1 glycans 1842:1848 large and sialylated glycans 1821:1848 large and sialylated glycans 1821:1848 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 9 19 theme antibody 2123:2130 arg1 interaction 2140:2150 the antibody binding interaction 2119:2150 the antibody binding interaction 2119:2150 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 6 20 theme glycan 1269:1274 arg1 profiles 1276:1283 The glycan profiles 1265:1283 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 1265:1332 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 3 21 dep receptors 668:676 arg1 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 (CD16a) 733:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 RIIb 716:719 RIIb 716:719 RIIb (CD32b) 716:727 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 CD16a 753:757 CD16a 753:757 CD16a 753:757 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 RIIaArg131/His131 689:705 RIIaArg131/His131 689:705 RIIaArg131/His131 (CD32a) 689:713 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 CD32b 722:726 CD32b 722:726 CD32b 722:726 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 receptors 668:676 human Fcγ receptors 658:676 human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 658:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 RI 678:679 RI (CD64) 678:686 RI (CD64) 678:686 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 21 dep receptors 668:676 arg1 CD32a 708:712 CD32a 708:712 CD32a 708:712 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 8 22 theme interaction 1933:1943 arg1 destabilization 1910:1924 destabilization 1910:1924 destabilization of the interaction 1910:1943 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 4 23 theme antibody 866:873 arg1 rituximab 875:883 the monoclonal antibody rituximab 851:883 the monoclonal antibody rituximab 851:883 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 5 24 theme binding 1140:1146 arg1 profiles 1148:1155 the binding profiles 1136:1155 the binding profiles of rituximab 1136:1168 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 8 25 theme rituximab 1876:1884 arg1 binding 1886:1892 rituximab binding 1876:1892 rituximab binding 1876:1892 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 3 26 theme glycans 788:794 arg1 role 776:779 the role 772:779 the role of the glycans in the binding mechanism with IgG 772:828 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 2 27 theme FcγRs 371:375 arg1 engagement 357:366 a productive molecular engagement 334:366 a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction 334:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 0 28 theme Glycoforms 36:45 arg1 Identification 0:13 Identification 0:13 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 0:103 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 6 29 theme FcγR 1600:1603 arg1 patterns 1613:1620 the respective FcγR binding patterns 1585:1620 the respective FcγR binding patterns with rituximab 1585:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 2 30 theme glycan 404:409 arg1 moiety 411:416 glycan moiety 404:416 glycan moiety of antibody and receptor 404:441 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 0 31 theme Fc 18:19 arg1 Glycoforms 36:45 Fc Gamma Receptor Glycoforms 18:45 Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab 18:102 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 1 32 theme gamma 108:112 arg1 FcγR 125:128 FcγR 125:128 FcγR 125:128 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 1 32 theme gamma 108:112 arg1 receptors 114:122 Fc gamma receptors 105:122 Fc gamma receptors (FcγR) 105:129 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 0 33 theme Differential 60:71 arg1 Kinetics 81:88 Differential Binding Kinetics 60:88 Differential Binding Kinetics for Rituximab 60:102 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 6 34 theme expressed 1292:1300 arg1 FcγRI 1302:1306 CHO expressed FcγRI 1288:1306 CHO expressed FcγRI 1288:1306 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 4 35 with interactions 1026:1037 arg1 HEK293 1051:1056 HEK293 1051:1056 HEK293 1051:1056 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 4 35 with interactions 1026:1037 arg1 murine 1063:1068 murine 1063:1068 murine 1063:1068 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 4 35 with interactions 1026:1037 arg1 human 1044:1048 human 1044:1048 human 1044:1048 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 7 36 theme rituximab 1745:1753 arg1 interaction 1755:1765 the high affinity rituximab interaction 1727:1765 the high affinity rituximab interaction through a stabilization effect 1727:1796 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 4 37 dep human 1044:1048 arg1 receptors 1085:1093 produced receptors 1076:1093 produced receptors 1076:1093 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 2 38 dep protein 392:398 arg1 the 388:390 the 388:390 the 388:390 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 7 39 theme high 1731:1734 arg1 interaction 1755:1765 the high affinity rituximab interaction 1727:1765 the high affinity rituximab interaction through a stabilization effect 1727:1796 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 3 40 theme Chinese 620:626 arg1 CHO 643:645 CHO 643:645 CHO 643:645 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 40 theme Chinese 620:626 arg1 ovary 636:640 Chinese hamster ovary 620:640 Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 620:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 6 41 theme receptors 1380:1388 arg1 profiles 1364:1371 the glycan profiles 1353:1371 the glycan profiles of the receptors expressed in NS0 and HEK293 cells 1353:1422 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 4 42 with interactions 835:846 arg1 FcγR 895:898 each FcγR 890:898 each FcγR 890:898 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 3 43 theme composition 605:615 arg1 description 579:589 a complete description 568:589 a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 568:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 5 44 theme cell-dependent 1230:1243 arg1 glycosylation 1250:1262 host cell-dependent FcγR glycosylation 1225:1262 host cell-dependent FcγR glycosylation 1225:1262 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 2 45 theme productive 336:345 arg1 engagement 357:366 a productive molecular engagement 334:366 a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction 334:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 1 46 theme cell-based 222:231 arg1 responses 240:248 antibody-dependent cell-based immune responses 203:248 antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP 203:275 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 1 46 theme cell-based 222:231 arg1 ADCC 258:261 ADCC 258:261 ADCC 258:261 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 6 47 theme glycan 1357:1362 arg1 profiles 1364:1371 the glycan profiles 1353:1371 the glycan profiles of the receptors expressed in NS0 and HEK293 cells 1353:1422 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 2 48 attach linked 324:329 arg1 engagement 357:366 a productive molecular engagement 334:366 a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction 334:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 2 48 attach linked 324:329 arg2 activity 303:310 The immune effector cell activity 278:310 The immune effector cell activity 278:310 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 2 49 from role 511:514 arg1 interaction 544:554 this interaction 539:554 this interaction 539:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 3 50 theme ovary 636:640 arg1 composition 605:615 the glycan composition 594:615 the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 594:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 9 51 from role 2095:2098 arg1 interaction 2140:2150 the antibody binding interaction 2119:2150 the antibody binding interaction 2119:2150 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 2 52 theme glycans 528:534 arg1 role 511:514 the role 507:514 the role of the FcγR glycans in this interaction 507:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 4 53 theme rituximab 875:883 arg1 interactions 835:846 The interactions 831:846 The interactions of the monoclonal antibody rituximab with each FcγR 831:898 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 3 54 theme Fcγ 664:666 arg1 receptors 668:676 human Fcγ receptors 658:676 human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 658:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 54 theme Fcγ 664:666 arg1 RIIb 716:719 RIIb 716:719 RIIb (CD32b) 716:727 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 54 theme Fcγ 664:666 arg1 RIIaArg131/His131 689:705 RIIaArg131/His131 689:705 RIIaArg131/His131 (CD32a) 689:713 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 54 theme Fcγ 664:666 arg1 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 733:750 RIIIaPhe158/Val158 (CD16a) 733:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 54 theme Fcγ 664:666 arg1 RI 678:679 RI (CD64) 678:686 RI (CD64) 678:686 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 3 55 from role 776:779 arg1 mechanism 811:819 the binding mechanism 799:819 the binding mechanism with IgG 799:828 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 2 56 theme effector 289:296 arg1 cell 298:301 The immune effector cell 278:301 The immune effector cell activity 278:310 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 8 57 theme large 1821:1825 arg1 glycans 1842:1848 large and sialylated glycans 1821:1848 large and sialylated glycans 1821:1848 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 6 58 theme respective 1589:1598 arg1 patterns 1613:1620 the respective FcγR binding patterns 1585:1620 the respective FcγR binding patterns with rituximab 1585:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 4 59 theme equivalent 1015:1024 arg1 interactions 1026:1037 the equivalent interactions 1011:1037 the equivalent interactions with human (HEK293) and murine (NS0) produced receptors 1011:1093 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 9 60 theme stabilizing 2083:2093 arg1 role 2095:2098 a stabilizing role 2081:2098 a stabilizing role of FcγR glycans in the antibody binding interaction 2081:2150 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 8 61 theme sialylated 1831:1840 arg1 glycans 1842:1848 large and sialylated glycans 1821:1848 large and sialylated glycans 1821:1848 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 1 62 theme prominent 185:193 arg1 role 195:198 a prominent role 183:198 a prominent role 183:198 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 5 63 from differences 1121:1131 arg1 profiles 1148:1155 the binding profiles 1136:1155 the binding profiles of rituximab 1136:1168 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 6 64 theme observed 1561:1568 arg1 differences 1570:1580 the observed differences 1557:1580 the observed differences in the respective FcγR binding patterns with rituximab 1557:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 9 65 theme binding 1994:2000 arg1 kinetics 2002:2009 the IgG1-FcγR binding kinetics 1980:2009 the IgG1-FcγR binding kinetics 1980:2009 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 4 66 theme produced 1076:1083 arg1 receptors 1085:1093 produced receptors 1076:1093 produced receptors 1076:1093 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 5 67 theme rituximab 1160:1168 arg1 profiles 1148:1155 the binding profiles 1136:1155 the binding profiles of rituximab 1136:1168 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 9 68 theme glycans 2108:2114 arg1 role 2095:2098 a stabilizing role 2081:2098 a stabilizing role of FcγR glycans in the antibody binding interaction 2081:2150 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 8 69 theme negative 1857:1864 arg1 impact 1866:1871 a negative impact 1855:1871 a negative impact 1855:1871 On FcγRI and FcγRIIIa large and sialylated glycans have a negative impact on rituximab binding, likely through destabilization of the interaction. 28576848 0 70 theme Binding 73:79 arg1 Kinetics 81:88 Differential Binding Kinetics 60:88 Differential Binding Kinetics for Rituximab 60:102 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 6 71 theme glycan 1445:1450 arg1 type 1452:1455 glycan type 1445:1455 glycan type 1445:1455 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 9 72 theme binding 2132:2138 arg1 interaction 2140:2150 the antibody binding interaction 2119:2150 the antibody binding interaction 2119:2150 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 1 73 theme Fc 140:141 arg1 region 143:148 the Fc region 136:148 the Fc region of antibodies 136:162 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 4 74 theme monoclonal 855:864 arg1 antibody 866:873 the monoclonal antibody 851:873 the monoclonal antibody rituximab 851:883 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 6 75 with patterns 1613:1620 arg1 rituximab 1627:1635 rituximab 1627:1635 rituximab 1627:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 6 76 theme glycan 1530:1535 arg1 differences 1537:1547 these glycan differences 1524:1547 these glycan differences 1524:1547 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 2 77 theme present 481:487 arg1 study 489:493 the present study 477:493 the present study 477:493 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 6 78 theme binding 1605:1611 arg1 patterns 1613:1620 the respective FcγR binding patterns 1585:1620 the respective FcγR binding patterns with rituximab 1585:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 1 79 theme antibody-dependent 203:220 arg1 responses 240:248 antibody-dependent cell-based immune responses 203:248 antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP 203:275 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 1 79 theme antibody-dependent 203:220 arg1 ADCC 258:261 ADCC 258:261 ADCC 258:261 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 6 80 from differences 1570:1580 arg1 patterns 1613:1620 the respective FcγR binding patterns 1585:1620 the respective FcγR binding patterns with rituximab 1585:1635 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 2 81 theme molecular 347:355 arg1 engagement 357:366 a productive molecular engagement 334:366 a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction 334:554 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 0 82 theme Gamma 21:25 arg1 Glycoforms 36:45 Fc Gamma Receptor Glycoforms 18:45 Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab 18:102 Identification of Fc Gamma Receptor Glycoforms That Produce Differential Binding Kinetics for Rituximab. 28576848 1 83 theme Fc 105:106 arg1 FcγR 125:128 FcγR 125:128 FcγR 125:128 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 1 83 theme Fc 105:106 arg1 receptors 114:122 Fc gamma receptors 105:122 Fc gamma receptors (FcγR) 105:129 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 6 84 theme FcγRI 1302:1306 arg1 profiles 1276:1283 The glycan profiles 1265:1283 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 1265:1332 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 3 85 theme binding 803:809 arg1 mechanism 811:819 the binding mechanism 799:819 the binding mechanism with IgG 799:828 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 2 86 theme receptor 434:441 arg1 moiety 411:416 glycan moiety 404:416 glycan moiety of antibody and receptor 404:441 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 2 86 theme receptor 434:441 arg1 protein 392:398 protein 392:398 protein 392:398 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 3 87 theme complete 570:577 arg1 description 579:589 a complete description 568:589 a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 568:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 6 88 theme CHO 1288:1290 arg1 FcγRI 1302:1306 CHO expressed FcγRI 1288:1306 CHO expressed FcγRI 1288:1306 The glycan profiles of CHO expressed FcγRI and FcγRIIIaPhe158/Val158 were compared with the glycan profiles of the receptors expressed in NS0 and HEK293 cells and we show that the glycan type and abundance differs significantly between the receptors and that these glycan differences lead to the observed differences in the respective FcγR binding patterns with rituximab. 28576848 7 89 theme stabilization 1777:1789 arg1 effect 1791:1796 a stabilization effect 1775:1796 a stabilization effect 1775:1796 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 2 90 theme antibody 421:428 arg1 moiety 411:416 glycan moiety 404:416 glycan moiety of antibody and receptor 404:441 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 2 90 theme antibody 421:428 arg1 protein 392:398 protein 392:398 protein 392:398 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 9 91 theme IgG1-FcγR 1984:1992 arg1 kinetics 2002:2009 the IgG1-FcγR binding kinetics 1980:2009 the IgG1-FcγR binding kinetics 1980:2009 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 7 92 theme affinity 1736:1743 arg1 interaction 1755:1765 the high affinity rituximab interaction 1727:1765 the high affinity rituximab interaction through a stabilization effect 1727:1796 Oligomannose structures are prevalent on FcγRI from each source and likely contribute to the high affinity rituximab interaction through a stabilization effect. 28576848 9 93 gly glycosylation 2035:2047 arg1 FcγR 2056:2059 the FcγR 2052:2059 the FcγR 2052:2059 In conclusion, the data show that the IgG1-FcγR binding kinetics differ depending on the glycosylation of the FcγR and further support a stabilizing role of FcγR glycans in the antibody binding interaction. 28576848 4 94 theme CHO-FcγRI 968:976 arg1 interactions 978:989 the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions 934:989 the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions 934:989 The interactions of the monoclonal antibody rituximab with each FcγR were characterized and we discuss the CHO-FcγRIIIaPhe158/Val158 and CHO-FcγRI interactions and compare them to the equivalent interactions with human (HEK293) and murine (NS0) produced receptors. 28576848 3 95 theme glycan 598:603 arg1 composition 605:615 the glycan composition 594:615 the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) 594:758 We provide a complete description of the glycan composition of Chinese hamster ovary (CHO) expressed human Fcγ receptors RI (CD64), RIIaArg131/His131 (CD32a), RIIb (CD32b) and RIIIaPhe158/Val158 (CD16a) and analyze the role of the glycans in the binding mechanism with IgG. 28576848 5 96 theme host 1225:1228 arg1 glycosylation 1250:1262 host cell-dependent FcγR glycosylation 1225:1262 host cell-dependent FcγR glycosylation 1225:1262 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 2 97 theme cell 298:301 arg1 activity 303:310 The immune effector cell activity 278:310 The immune effector cell activity 278:310 The immune effector cell activity is directly linked to a productive molecular engagement of FcγRs where both the protein and glycan moiety of antibody and receptor can affect the interaction and in the present study we focus on the role of the FcγR glycans in this interaction. 28576848 5 98 theme clear 1115:1119 arg1 differences 1121:1131 clear differences 1115:1131 clear differences 1115:1131 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 5 99 theme FcγR 1245:1248 arg1 glycosylation 1250:1262 host cell-dependent FcγR glycosylation 1225:1262 host cell-dependent FcγR glycosylation 1225:1262 Our results reveal clear differences in the binding profiles of rituximab, which we attribute in each case to the differences in host cell-dependent FcγR glycosylation. 28576848 1 100 theme immune 233:238 arg1 responses 240:248 antibody-dependent cell-based immune responses 203:248 antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP 203:275 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28576848 1 100 theme immune 233:238 arg1 ADCC 258:261 ADCC 258:261 ADCC 258:261 Fc gamma receptors (FcγR) bind the Fc region of antibodies and therefore play a prominent role in antibody-dependent cell-based immune responses such as ADCC, CDC and ADCP. 28109441 3 0 theme comprehensive 524:536 arg1 characterization 538:553 comprehensive characterization 524:553 comprehensive characterization of protein glycosylation 524:578 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 5 1 from samples 796:802 arg1 purification 756:767 purification 756:767 purification 756:767 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 5 1 from samples 796:802 arg1 release 744:750 release 744:750 release 744:750 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 6 2 theme effective 814:822 arg1 methods 833:839 The most effective labeling methods 805:839 The most effective labeling methods developed for sensitive quantitative glycomics 805:886 The most effective labeling methods developed for sensitive quantitative glycomics are also described and discussed. 28109441 7 3 used used 968:971 arg2 approaches 942:951 The chromatographic approaches 922:951 The chromatographic approaches that have been used effectively in glycomics 922:996 The chromatographic approaches that have been used effectively in glycomics are also highlighted. 28109441 2 4 theme turnover 340:347 arg1 rates 349:353 protein turnover rates 332:353 protein turnover rates 332:353 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 5 5 dep release 744:750 arg1 the 740:742 the 740:742 the 740:742 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 2 6 theme mechanisms 293:302 arg1 recognition 247:257 both direct recognition 235:257 both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation 235:382 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 3 7 from attributes 400:409 arg1 implications 459:470 implications 459:470 implications in a number of diseases 459:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 3 7 from attributes 400:409 arg1 processes 445:453 numerous biological processes 425:453 numerous biological processes 425:453 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 2 8 theme indirect 284:291 arg1 mechanisms 293:302 indirect mechanisms 284:302 indirect mechanisms 284:302 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 2 9 theme protein 332:338 arg1 rates 349:353 protein turnover rates 332:353 protein turnover rates 332:353 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 0 10 theme Quantitative 0:11 arg1 Glycomics 13:21 Quantitative Glycomics 0:21 Quantitative Glycomics 0:21 Quantitative Glycomics: A Combined Analytical and Bioinformatics Approach. 28109441 4 11 theme cutting-edge 602:613 arg1 methods 615:621 cutting-edge methods 602:621 cutting-edge methods 602:621 This chapter reviews cutting-edge methods and tools developed to facilitate quantitative glycomics. 28109441 5 12 theme different 709:717 arg1 methods 719:725 the different methods 705:725 the different methods employed for the release and purification of glycans from biological samples 705:802 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 6 13 theme quantitative 865:876 arg1 glycomics 878:886 sensitive quantitative glycomics 855:886 sensitive quantitative glycomics 855:886 The most effective labeling methods developed for sensitive quantitative glycomics are also described and discussed. 28109441 7 14 theme chromatographic 926:940 arg1 approaches 942:951 The chromatographic approaches 922:951 The chromatographic approaches that have been used effectively in glycomics 922:996 The chromatographic approaches that have been used effectively in glycomics are also highlighted. 28109441 5 15 theme glycans 772:778 arg1 purification 756:767 purification 756:767 purification 756:767 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 5 15 theme glycans 772:778 arg1 release 744:750 release 744:750 release 744:750 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 6 16 theme sensitive 855:863 arg1 glycomics 878:886 sensitive quantitative glycomics 855:886 sensitive quantitative glycomics 855:886 The most effective labeling methods developed for sensitive quantitative glycomics are also described and discussed. 28109441 3 17 theme biological 389:398 arg1 attributes 400:409 The biological attributes 385:409 The biological attributes of glycans in numerous biological processes and implications in a number of diseases 385:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 3 18 theme glycosylation 566:578 arg1 characterization 538:553 comprehensive characterization 524:553 comprehensive characterization of protein glycosylation 524:578 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 3 19 theme diseases 487:494 arg1 number 477:482 a number 475:482 a number of diseases 475:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 3 20 theme numerous 425:432 arg1 processes 445:453 numerous biological processes 425:453 numerous biological processes 425:453 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 5 21 theme biological 785:794 arg1 samples 796:802 biological samples 785:802 biological samples 785:802 This chapter highlights the different methods employed for the release and purification of glycans from biological samples. 28109441 0 22 theme Combined 26:33 arg1 Analytical 35:44 A Combined Analytical and Bioinformatics Approach 24:72 Analytical 35:44 Quantitative Glycomics: A Combined Analytical and Bioinformatics Approach. 28109441 3 23 theme biological 434:443 arg1 processes 445:453 numerous biological processes 425:453 numerous biological processes 425:453 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 2 24 theme direct 240:245 arg1 recognition 247:257 both direct recognition 235:257 both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation 235:382 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 3 25 theme glycans 414:420 arg1 attributes 400:409 The biological attributes 385:409 The biological attributes of glycans in numerous biological processes and implications in a number of diseases 385:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 0 26 dep Analytical 35:44 arg1 Glycomics 13:21 Quantitative Glycomics 0:21 Quantitative Glycomics 0:21 Quantitative Glycomics: A Combined Analytical and Bioinformatics Approach. 28109441 1 27 theme common 108:113 arg1 modifications 137:149 the most common and essential protein modifications 99:149 the most common and essential protein modifications 99:149 Glycosylation is one of the most common and essential protein modifications. 28109441 4 28 theme quantitative 657:668 arg1 glycomics 670:678 quantitative glycomics 657:678 quantitative glycomics 657:678 This chapter reviews cutting-edge methods and tools developed to facilitate quantitative glycomics. 28109441 2 29 theme molecules 217:225 arg1 function 200:207 the function 196:207 the function of such molecules 196:225 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 2 30 theme such 212:215 arg1 molecules 217:225 such molecules 212:225 such molecules 212:225 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 6 31 theme labeling 824:831 arg1 methods 833:839 The most effective labeling methods 805:839 The most effective labeling methods developed for sensitive quantitative glycomics 805:886 The most effective labeling methods developed for sensitive quantitative glycomics are also described and discussed. 28109441 1 32 theme essential 119:127 arg1 modifications 137:149 the most common and essential protein modifications 99:149 the most common and essential protein modifications 99:149 Glycosylation is one of the most common and essential protein modifications. 28109441 2 33 theme structures 269:278 arg1 recognition 247:257 both direct recognition 235:257 both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation 235:382 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 3 34 from processes 445:453 arg1 number 477:482 a number 475:482 a number of diseases 475:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 1 35 theme protein 129:135 arg1 modifications 137:149 the most common and essential protein modifications 99:149 the most common and essential protein modifications 99:149 Glycosylation is one of the most common and essential protein modifications. 28109441 2 36 theme rates 349:353 arg1 control 321:327 the control 317:327 the control of protein turnover rates 317:353 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 2 36 theme rates 349:353 arg1 stability 356:364 stability 356:364 stability 356:364 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 2 36 theme rates 349:353 arg1 conformation 371:382 conformation 371:382 conformation 371:382 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 2 37 theme glycan 262:267 arg1 structures 269:278 glycan structures 262:278 glycan structures 262:278 Glycans conjugated to biomolecules modulate the function of such molecules through both direct recognition of glycan structures and indirect mechanisms that involve the control of protein turnover rates, stability, and conformation. 28109441 1 38 theme modifications 137:149 arg1 modifications 137:149 the most common and essential protein modifications 99:149 the most common and essential protein modifications 99:149 Glycosylation is one of the most common and essential protein modifications. 28109441 1 38 theme modifications 137:149 arg1 one 92:94 one 92:94 one 92:94 Glycosylation is one of the most common and essential protein modifications. 28109441 0 39 theme Bioinformatics 50:63 arg1 Approach 65:72 A Combined Analytical and Bioinformatics Approach 24:72 Approach 65:72 Quantitative Glycomics: A Combined Analytical and Bioinformatics Approach. 28109441 3 40 from implications 459:470 arg1 number 477:482 a number 475:482 a number of diseases 475:494 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 28109441 3 41 theme protein 558:564 arg1 glycosylation 566:578 protein glycosylation 558:578 protein glycosylation 558:578 The biological attributes of glycans in numerous biological processes and implications in a number of diseases highlight the necessity for comprehensive characterization of protein glycosylation. 25849464 7 0 theme isolated 1102:1109 arg1 gene 1116:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 1 theme kinetic 1732:1738 arg1 results 1740:1746 the kinetic results 1728:1746 the kinetic results 1728:1746 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 11 2 theme new 1921:1923 arg1 insights 1925:1932 new insights 1921:1932 new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase 1921:2027 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 10 3 theme local 1769:1773 arg1 networks 1782:1789 the local H-bond networks 1765:1789 the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties 1765:1854 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 5 4 theme ± 694:694 arg1 μM 700:701 65.0 ± 6.5 μM 689:701 65.0 ± 6.5 μM 689:701 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 2 5 attach isolated 312:319 arg2 laccase 285:291 a native laccase 276:291 a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 276:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 2 5 attach isolated 312:319 arg1 sp 354:355 a white-rot fungus Lentinus sp 326:355 a white-rot fungus Lentinus sp 326:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 2 5 attach isolated 312:319 arg2 nLcc4 305:309 designated nLcc4 294:309 designated nLcc4 294:309 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 9 6 theme cultural 1517:1524 arg1 media 1526:1530 the cultural media 1513:1530 the cultural media of P. pastoris cells 1513:1551 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 6 7 theme kinetic 1039:1045 arg1 efficiency 1047:1056 similar kinetic efficiency 1031:1056 similar kinetic efficiency 1031:1056 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 5 8 theme rate 582:585 arg1 analysis 595:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 9 theme lignosulfonic 754:766 arg1 acid 768:771 lignosulfonic acid 754:771 lignosulfonic acid 754:771 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 11 10 from role 1943:1946 arg1 function 1986:1993 function 1986:1993 function 1986:1993 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 11 10 from role 1943:1946 arg1 structure 1972:1980 structure 1972:1980 structure 1972:1980 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 7 11 with frame 1146:1150 arg1 polypeptide 1178:1188 a deduced polypeptide 1168:1188 a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1168:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 2 12 theme laccase 285:291 arg1 structure 243:251 the crystal structure 231:251 the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 231:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 9 13 theme pastoris 1538:1545 arg1 cells 1547:1551 P. pastoris cells 1535:1551 P. pastoris cells 1535:1551 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 8 14 theme Recombinant 1291:1301 arg1 Lcc4 1313:1316 Recombinant wild-type Lcc4 1291:1316 Recombinant wild-type Lcc4 1291:1316 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 7 15 theme predicted 1229:1237 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 7 16 theme deduced 1170:1176 arg1 polypeptide 1178:1188 a deduced polypeptide 1168:1188 a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1168:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 17 theme dynamics 1642:1649 arg1 simulations 1651:1661 Molecular dynamics simulations 1632:1661 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc 1632:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 4 18 gly N-glycosylated 507:520 arg1 sites 522:526 three N-glycosylated sites 501:526 three N-glycosylated sites at Asn75, Asn238, and Asn458 501:555 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 5 19 theme 0.234 828:832 arg1 s-1 834:836 0.234 s-1 828:836 0.234 s-1 828:836 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 7 20 from peptide 1249:1255 arg1 N-terminus 1279:1288 the N-terminus 1275:1288 the N-terminus 1275:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 8 21 theme mutant 1322:1327 arg1 enzymes 1329:1335 mutant enzymes 1322:1335 mutant enzymes 1322:1335 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 7 22 theme acid 1203:1206 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 7 22 theme acid 1203:1206 arg1 residues 1208:1215 521 amino acid residues 1193:1215 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1193:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 23 theme various 1675:1681 arg1 states 1683:1688 various states 1675:1688 various states of (de-)glycosylation in nLcc 1675:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 5 24 theme isothermal 790:799 arg1 calorimetry 811:821 isothermal titration calorimetry 790:821 isothermal titration calorimetry 790:821 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 6 25 theme H-deglycosylated 892:907 arg1 nLcc4 909:913 Endo H-deglycosylated nLcc4 887:913 Endo H-deglycosylated nLcc4 (dLcc4) 887:921 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 6 25 theme H-deglycosylated 892:907 arg1 dLcc4 916:920 dLcc4 916:920 dLcc4 916:920 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 11 26 theme laccase 2021:2027 arg1 function 1986:1993 function 1986:1993 function 1986:1993 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 11 26 theme laccase 2021:2027 arg1 structure 1972:1980 structure 1972:1980 structure 1972:1980 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 4 27 theme T1 463:464 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 0 28 theme Lentinus 72:79 arg1 sp 81:82 Lentinus sp 72:82 Lentinus sp 72:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 7 29 contain contains 1121:1128 arg1 gene 1116:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 7 29 contain contains 1121:1128 arg2 frame 1146:1150 an open reading frame 1130:1150 an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1130:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 1 30 theme organic 159:165 arg1 compounds 181:189 various organic and inorganic compounds 151:189 various organic and inorganic compounds 151:189 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 7 31 from N-terminus 1279:1288 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 7 31 from N-terminus 1279:1288 arg1 residues 1263:1270 21 residues 1260:1270 21 residues at the N-terminus 1260:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 4 32 theme T2/T3 470:474 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 9 33 theme wild-type 1613:1621 arg1 laccase 1623:1629 the wild-type laccase 1609:1629 the wild-type laccase 1609:1629 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 8 34 theme Pichia 1377:1382 arg1 pastoris 1384:1391 Pichia pastoris 1377:1391 Pichia pastoris 1377:1391 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 3 35 theme cupredoxin-like 385:399 arg1 domains 401:407 three cupredoxin-like domains D1-D3 379:413 three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology 379:460 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 0 36 theme Structural 0:9 arg1 roles 26:30 Structural and functional roles 0:30 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp 0:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 5 37 theme ± 844:844 arg1 μM 850:851 56.7 ± 3.2 μM 839:851 56.7 ± 3.2 μM 839:851 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 38 with Km 628:629 arg1 ABTS 668:671 substrate ABTS 658:671 substrate ABTS 658:671 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 4 39 theme N-glycosylated 507:520 arg1 sites 522:526 three N-glycosylated sites 501:526 three N-glycosylated sites at Asn75, Asn238, and Asn458 501:555 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 5 40 dep using 784:788 arg1 μM 850:851 56.7 ± 3.2 μM 839:851 56.7 ± 3.2 μM 839:851 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 40 dep using 784:788 arg1 s-1 834:836 0.234 s-1 828:836 0.234 s-1 828:836 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 40 dep using 784:788 arg1 s-1μM-1 864:870 0.004 s-1μM-1 858:870 0.004 s-1μM-1 858:870 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 0 41 theme functional 15:24 arg1 roles 26:30 Structural and functional roles 0:30 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp 0:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 1 42 theme multi-copper 98:109 arg1 Laccases 85:92 Laccases 85:92 Laccases 85:92 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 1 42 theme multi-copper 98:109 arg1 oxidases 111:118 multi-copper oxidases 98:118 multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water 98:213 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 10 43 from states 1683:1688 arg1 nLcc 1715:1718 nLcc 1715:1718 nLcc 1715:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 11 44 theme Basidiomycete 2000:2012 arg1 laccase 2021:2027 a Basidiomycete fungal laccase 1998:2027 a Basidiomycete fungal laccase 1998:2027 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 10 45 theme de- 1694:1696 arg1 glycosylation 1698:1710 (de-)glycosylation 1693:1710 (de-)glycosylation in nLcc 1693:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 3 46 dep domains 401:407 arg1 D1-D3 409:413 D1-D3 409:413 three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology 379:460 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 3 46 dep domains 401:407 arg1 each 415:418 each 415:418 each 415:418 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 4 47 theme binding 483:489 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 8 48 theme enzyme 1425:1430 arg1 activity 1432:1439 enzyme activity 1425:1439 enzyme activity 1425:1439 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 6 49 from sites 1000:1004 arg1 enzyme 1013:1018 the enzyme 1009:1018 the enzyme 1009:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 3 50 theme key 440:442 arg1 topology 453:460 a Greek key β-barrel topology 432:460 a Greek key β-barrel topology 432:460 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 7 51 theme open 1133:1136 arg1 frame 1146:1150 an open reading frame 1130:1150 an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1130:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 52 theme glycosylation 1698:1710 arg1 states 1683:1688 various states 1675:1688 various states of (de-)glycosylation in nLcc 1675:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 2 53 theme fungus 338:343 arg1 sp 354:355 a white-rot fungus Lentinus sp 326:355 a white-rot fungus Lentinus sp 326:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 1 54 theme inorganic 171:179 arg1 compounds 181:189 various organic and inorganic compounds 151:189 various organic and inorganic compounds 151:189 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 9 55 theme mutant 1486:1491 arg1 enzymes 1493:1499 The mutant enzymes 1482:1499 The mutant enzymes secreted in the cultural media of P. pastoris cells 1482:1551 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 5 56 with kcat 622:625 arg1 ABTS 668:671 substrate ABTS 658:671 substrate ABTS 658:671 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 8 57 theme glycosylation 1451:1463 arg1 site 1465:1468 single glycosylation site 1444:1468 single glycosylation site deficiency 1444:1479 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 11 58 theme glycosylation 1951:1963 arg1 role 1943:1946 the role 1939:1946 the role of glycosylation in the structure and function of a Basidiomycete fungal laccase 1939:2027 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 7 59 theme Lcc4 1111:1114 arg1 gene 1116:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene 1098:1119 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 60 from glycosylation 1698:1710 arg1 nLcc 1715:1718 nLcc 1715:1718 nLcc 1715:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 5 61 theme titration 801:809 arg1 calorimetry 811:821 isothermal titration calorimetry 790:821 isothermal titration calorimetry 790:821 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 62 theme Initial 574:580 arg1 analysis 595:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 2 63 theme Å 260:260 arg1 resolution 262:271 1.8 Å resolution 256:271 1.8 Å resolution 256:271 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 10 64 gly glycosylation 1698:1710 arg1 nLcc 1715:1718 nLcc 1715:1718 nLcc 1715:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 10 65 from nLcc 1715:1718 arg1 states 1683:1688 various states 1675:1688 various states of (de-)glycosylation in nLcc 1675:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 6 66 theme similar 1031:1037 arg1 efficiency 1047:1056 similar kinetic efficiency 1031:1056 similar kinetic efficiency 1031:1056 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 4 67 from Asn458 550:555 arg1 sites 522:526 three N-glycosylated sites 501:526 three N-glycosylated sites at Asn75, Asn238, and Asn458 501:555 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 4 67 from Asn458 550:555 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 5 68 theme kinetic 587:593 arg1 analysis 595:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis 574:602 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 1 69 theme compounds 181:189 arg1 oxidation 138:146 the oxidation 134:146 the oxidation of various organic and inorganic compounds 134:189 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 8 70 from effect 1415:1420 arg1 activity 1432:1439 enzyme activity 1425:1439 enzyme activity 1425:1439 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 7 71 theme residues 1263:1270 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 2 72 theme native 278:283 arg1 laccase 285:291 a native laccase 276:291 a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 276:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 2 72 theme native 278:283 arg1 nLcc4 305:309 designated nLcc4 294:309 designated nLcc4 294:309 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 9 73 theme cells 1547:1551 arg1 media 1526:1530 the cultural media 1513:1530 the cultural media of P. pastoris cells 1513:1551 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 0 74 from sp 81:82 arg1 laccase 59:65 fungal laccase 52:65 fungal laccase from Lentinus sp 52:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 0 74 from sp 81:82 arg1 roles 26:30 Structural and functional roles 0:30 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp 0:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 10 75 theme H-bond 1775:1780 arg1 networks 1782:1789 the local H-bond networks 1765:1789 the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties 1765:1854 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 5 76 theme nLcc4 647:651 arg1 s-1 684:686 3,382 s-1 678:686 3,382 s-1 678:686 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 76 theme nLcc4 647:651 arg1 Km 628:629 Km 628:629 Km 628:629 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 76 theme nLcc4 647:651 arg1 kcat/Km 636:642 kcat/Km 636:642 kcat/Km 636:642 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 5 76 theme nLcc4 647:651 arg1 kcat 622:625 the kcat 618:625 the kcat 618:625 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 9 77 theme P. 1535:1536 arg1 cells 1547:1551 P. pastoris cells 1535:1551 P. pastoris cells 1535:1551 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 10 78 theme simulations 1651:1661 arg1 analyses 1663:1670 Molecular dynamics simulations analyses 1632:1670 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc 1632:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 10 79 theme crucial 1863:1869 arg1 role 1871:1874 a crucial role 1861:1874 a crucial role 1861:1874 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 7 80 theme signaling 1239:1247 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 81 theme loop 1821:1824 arg1 D2-D3 1826:1830 loop D2-D3 1821:1830 loop D2-D3 1821:1830 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 10 82 theme Molecular 1632:1640 arg1 simulations 1651:1661 Molecular dynamics simulations 1632:1661 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc 1632:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 10 83 theme states 1683:1688 arg1 analyses 1663:1670 Molecular dynamics simulations analyses 1632:1670 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc 1632:1718 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 7 84 theme residues 1208:1215 arg1 polypeptide 1178:1188 a deduced polypeptide 1168:1188 a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1168:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 10 85 theme glycan 1840:1845 arg1 moieties 1847:1854 the glycan moieties 1836:1854 the glycan moieties 1836:1854 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 2 86 theme crystal 235:241 arg1 structure 243:251 the crystal structure 231:251 the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 231:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 2 87 from resolution 262:271 arg1 structure 243:251 the crystal structure 231:251 the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 231:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 0 88 from roles 26:30 arg1 laccase 59:65 fungal laccase 52:65 fungal laccase from Lentinus sp 52:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 0 88 from roles 26:30 arg1 sp 81:82 Lentinus sp 72:82 Lentinus sp 72:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 7 89 theme amino 1197:1201 arg1 peptide 1249:1255 a predicted signaling peptide 1227:1255 a predicted signaling peptide of 21 residues at the N-terminus 1227:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 7 89 theme amino 1197:1201 arg1 residues 1208:1215 521 amino acid residues 1193:1215 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1193:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 1 90 theme various 151:157 arg1 compounds 181:189 various organic and inorganic compounds 151:189 various organic and inorganic compounds 151:189 Laccases are multi-copper oxidases that catalyze the oxidation of various organic and inorganic compounds by reducing O2 to water. 25849464 6 91 theme Endo 887:890 arg1 nLcc4 909:913 Endo H-deglycosylated nLcc4 887:913 Endo H-deglycosylated nLcc4 (dLcc4) 887:921 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 6 91 theme Endo 887:890 arg1 dLcc4 916:920 dLcc4 916:920 dLcc4 916:920 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 9 92 theme activity 1597:1604 arg1 % 1588:1588 only 4-50% 1579:1588 only 4-50% of the activity of the wild-type laccase 1579:1629 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 9 92 theme activity 1597:1604 arg1 activity 1597:1604 the activity 1593:1604 the activity of the wild-type laccase 1593:1629 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 4 93 theme copper 476:481 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 8 94 theme wild-type 1303:1311 arg1 Lcc4 1313:1316 Recombinant wild-type Lcc4 1291:1316 Recombinant wild-type Lcc4 1291:1316 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 5 95 theme substrate 658:666 arg1 ABTS 668:671 substrate ABTS 658:671 substrate ABTS 658:671 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 2 96 theme designated 294:303 arg1 laccase 285:291 a native laccase 276:291 a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp 276:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 2 96 theme designated 294:303 arg1 nLcc4 305:309 designated nLcc4 294:309 designated nLcc4 294:309 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 4 97 from Asn238 538:543 arg1 sites 522:526 three N-glycosylated sites 501:526 three N-glycosylated sites at Asn75, Asn238, and Asn458 501:555 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 4 97 from Asn238 538:543 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 11 98 gly glycosylation 1951:1963 arg1 laccase 2021:2027 a Basidiomycete fungal laccase 1998:2027 a Basidiomycete fungal laccase 1998:2027 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 6 99 theme remaining 953:961 arg1 residue 945:951 only one GlcNAc residue remaining 929:961 only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme 929:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 9 100 theme laccase 1623:1629 arg1 activity 1597:1604 the activity 1593:1604 the activity of the wild-type laccase 1593:1629 The mutant enzymes secreted in the cultural media of P. pastoris cells were observed to maintain only 4-50% of the activity of the wild-type laccase. 25849464 4 101 from Asn75 531:535 arg1 sites 522:526 three N-glycosylated sites 501:526 three N-glycosylated sites at Asn75, Asn238, and Asn458 501:555 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 4 101 from Asn75 531:535 arg1 sites 491:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites 463:495 T1 and T2/T3 copper binding sites and three N-glycosylated sites at Asn75, Asn238, and Asn458 were elucidated. 25849464 6 102 theme GlcNAc 938:943 arg1 residue 945:951 only one GlcNAc residue remaining 929:961 only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme 929:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 6 103 gly N-glycosylation 984:998 arg2 sites 1000:1004 the three N-glycosylation sites 974:1004 the three N-glycosylation sites in the enzyme 974:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 6 103 gly N-glycosylation 984:998 arg2 three 978:982 three 978:982 three 978:982 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 2 104 theme Lentinus 345:352 arg1 sp 354:355 a white-rot fungus Lentinus sp 326:355 a white-rot fungus Lentinus sp 326:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 25849464 6 105 with nLcc4 909:913 arg1 residue 945:951 only one GlcNAc residue remaining 929:961 only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme 929:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 7 106 theme bp 1160:1161 arg1 frame 1146:1150 an open reading frame 1130:1150 an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1130:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 5 107 with values 742:747 arg1 acid 768:771 lignosulfonic acid 754:771 lignosulfonic acid 754:771 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 3 108 theme Greek 434:438 arg1 topology 453:460 a Greek key β-barrel topology 432:460 a Greek key β-barrel topology 432:460 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 0 109 theme glycosylation 35:47 arg1 roles 26:30 Structural and functional roles 0:30 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp 0:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 6 110 theme thermal 1062:1068 arg1 stability 1070:1078 thermal stability 1062:1078 thermal stability 1062:1078 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 5 111 with kcat/Km 636:642 arg1 ABTS 668:671 substrate ABTS 658:671 substrate ABTS 658:671 Initial rate kinetic analysis revealed that the kcat, Km, and kcat/Km of nLcc4 with substrate ABTS were 3,382 s-1, 65.0 ± 6.5 μM, and 52 s-1μM-1, respectively; and the values with lignosulfonic acid determined using isothermal titration calorimetry were 0.234 s-1, 56.7 ± 3.2 μM, and 0.004 s-1μM-1, respectively. 25849464 11 112 theme fungal 2014:2019 arg1 laccase 2021:2027 a Basidiomycete fungal laccase 1998:2027 a Basidiomycete fungal laccase 1998:2027 This study provides new insights into the role of glycosylation in the structure and function of a Basidiomycete fungal laccase. 25849464 0 113 theme fungal 52:57 arg1 laccase 59:65 fungal laccase 52:65 fungal laccase from Lentinus sp 52:82 Structural and functional roles of glycosylation in fungal laccase from Lentinus sp. 25849464 8 114 dep Lcc4 1313:1316 arg1 N458D 1353:1357 N458D 1353:1357 N458D 1353:1357 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 8 114 dep Lcc4 1313:1316 arg1 N238D 1343:1347 N238D 1343:1347 N238D 1343:1347 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 8 115 gly glycosylation 1451:1463 arg2 site 1465:1468 single glycosylation site 1444:1468 single glycosylation site deficiency 1444:1479 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 7 116 theme reading 1138:1144 arg1 frame 1146:1150 an open reading frame 1130:1150 an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus 1130:1288 The isolated Lcc4 gene contains an open reading frame of 1563 bp with a deduced polypeptide of 521 amino acid residues including a predicted signaling peptide of 21 residues at the N-terminus. 25849464 6 117 theme N-glycosylation 984:998 arg1 sites 1000:1004 the three N-glycosylation sites 974:1004 the three N-glycosylation sites in the enzyme 974:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 10 118 theme laccase 1883:1889 arg1 activity 1891:1898 the laccase activity 1879:1898 the laccase activity 1879:1898 Molecular dynamics simulations analyses of various states of (de-)glycosylation in nLcc support the kinetic results and suggest that the local H-bond networks between the domain connecting loop D2-D3 and the glycan moieties play a crucial role in the laccase activity. 25849464 8 119 theme single 1444:1449 arg1 site 1465:1468 single glycosylation site 1444:1468 single glycosylation site deficiency 1444:1479 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 6 120 from each 966:969 arg1 residue 945:951 only one GlcNAc residue remaining 929:961 only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme 929:1018 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 3 121 theme β-barrel 444:451 arg1 topology 453:460 a Greek key β-barrel topology 432:460 a Greek key β-barrel topology 432:460 nLcc4 is composed of three cupredoxin-like domains D1-D3 each folded into a Greek key β-barrel topology. 25849464 8 122 theme site 1465:1468 arg1 deficiency 1470:1479 single glycosylation site deficiency 1444:1479 single glycosylation site deficiency 1444:1479 Recombinant wild-type Lcc4 and mutant enzymes N75D, N238D and N458D were expressed in Pichia pastoris cells to evaluate the effect on enzyme activity by single glycosylation site deficiency. 25849464 6 123 gly H-deglycosylated 892:907 arg1 nLcc4 909:913 Endo H-deglycosylated nLcc4 887:913 Endo H-deglycosylated nLcc4 (dLcc4) 887:921 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 6 123 gly H-deglycosylated 892:907 arg1 dLcc4 916:920 dLcc4 916:920 dLcc4 916:920 Endo H-deglycosylated nLcc4 (dLcc4), with only one GlcNAc residue remaining at each of the three N-glycosylation sites in the enzyme, exhibited similar kinetic efficiency and thermal stability to that of nLcc4. 25849464 2 124 theme white-rot 328:336 arg1 sp 354:355 a white-rot fungus Lentinus sp 326:355 a white-rot fungus Lentinus sp 326:355 Here we report the crystal structure at 1.8 Å resolution of a native laccase (designated nLcc4) isolated from a white-rot fungus Lentinus sp. 26255982 6 0 theme m/z 904:906 arg1 529.75 908:913 m/z 529.75 904:913 m/z 529.75 904:913 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 7 1 theme O-glycans 1080:1088 arg1 abundance 1063:1071 the abundance 1059:1071 the abundance of the O-glycans 1059:1088 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 5 2 theme LC-MS 578:582 arg1 data 584:587 the LC-MS data 574:587 the LC-MS data 574:587 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 1 3 theme cancer 174:179 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 3 theme cancer 174:179 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 9 4 theme O-glycan 1260:1267 arg1 structures 1269:1278 The O-glycan structures 1256:1278 The O-glycan structures 1256:1278 The O-glycan structures are increased in the serum of GC, they may be candidates for carbohydrate tumor markers. 26255982 9 5 theme tumor 1354:1358 arg1 markers 1360:1366 carbohydrate tumor markers 1341:1366 carbohydrate tumor markers 1341:1366 The O-glycan structures are increased in the serum of GC, they may be candidates for carbohydrate tumor markers. 26255982 0 6 theme gastric 99:105 arg1 cancer 107:112 gastric cancer 99:112 gastric cancer 99:112 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 6 7 theme m/z 841:843 arg1 809.42 845:850 m/z 809.42 841:850 m/z 809.42 841:850 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 7 theme m/z 841:843 arg1 m/z 829:831 m/z 733.33 829:838 m/z 733.33 829:838 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 5 8 theme Student 640:646 arg1 test 650:653 unpaired Student t test 631:653 unpaired Student t test 631:653 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 6 9 theme complex 810:816 arg1 O-glycans 818:826 core2 complex O-glycans 804:826 core2 complex O-glycans (m/z 733.33, m/z 809.42) 804:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 4 10 dep DESIGN 427:432 arg1 used 437:440 used 437:440 used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis 437:561 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 3 11 from differences 310:320 arg1 O-glycosylation 325:339 O-glycosylation 325:339 O-glycosylation 325:339 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 3 11 from differences 310:320 arg1 serum 348:352 the serum 344:352 the serum of 157 gastric cancer patients (GC) and 144 healthy donors 344:411 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 7 12 theme GC 1136:1137 arg1 serum 1127:1131 the serum 1123:1131 the serum of GC 1123:1137 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 4 13 theme LC-MS 548:552 arg1 analysis 554:561 LC-MS analysis 548:561 LC-MS analysis 548:561 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 5 14 theme O-glycans 700:708 arg1 information 685:695 the structural information 670:695 the structural information of O-glycans from LC-MS/MS in positive mode 670:739 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 4 15 theme EXPERIMENTAL 414:425 arg1 DESIGN 427:432 EXPERIMENTAL DESIGN 414:432 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 414:562 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 8 16 with patients 1238:1245 arg1 GC 1252:1253 GC 1252:1253 GC 1252:1253 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 3 17 theme patients 376:383 arg1 serum 348:352 the serum 344:352 the serum of 157 gastric cancer patients (GC) and 144 healthy donors 344:411 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 6 18 theme expression 754:763 arg1 level 765:769 The expression level 750:769 The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) 750:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 19 theme GC 963:964 arg1 serum 954:958 the serum 950:958 the serum of GC 950:964 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 20 theme O-glycans 818:826 arg1 level 765:769 The expression level 750:769 The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) 750:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 8 21 theme multiple 1206:1213 arg1 antigens 1225:1232 multiple candidate antigens 1206:1232 multiple candidate antigens for patients with GC 1206:1253 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 5 22 theme squares 606:612 arg1 discriminant 614:625 partial least squares discriminant 592:625 partial least squares discriminant 592:625 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 8 23 theme Glycomics 1174:1182 arg1 approaches 1184:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 3 24 theme donors 406:411 arg1 serum 348:352 the serum 344:352 the serum of 157 gastric cancer patients (GC) and 144 healthy donors 344:411 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 3 25 theme gastric 361:367 arg1 GC 386:387 GC 386:387 GC 386:387 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 3 25 theme gastric 361:367 arg1 patients 376:383 157 gastric cancer patients 357:383 157 gastric cancer patients (GC) 357:388 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 7 26 dep MUC1 1109:1112 arg1 CEA 1115:1117 CEA 1115:1117 CEA 1115:1117 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 6 27 theme core2 804:808 arg1 O-glycans 818:826 core2 complex O-glycans 804:826 core2 complex O-glycans (m/z 733.33, m/z 809.42) 804:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 4 28 attach released 476:483 arg1 proteins 490:497 proteins 490:497 proteins 490:497 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 4 28 attach released 476:483 arg2 O-glycans 465:473 labeling O-glycans 456:473 labeling O-glycans (released from proteins) 456:498 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 0 29 theme Liquid 0:5 arg1 chromatography 7:20 Liquid chromatography 0:20 Liquid chromatography mass spectrometry-based O-glycomics 0:56 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 5 30 theme structural 674:683 arg1 information 685:695 the structural information 670:695 the structural information of O-glycans from LC-MS/MS in positive mode 670:739 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 1 31 theme PURPOSE 115:121 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 31 theme PURPOSE 115:121 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 6 32 dep RESULTS 742:748 arg1 increased 858:866 increased 858:866 were increased significantly (p < 0.0001) 853:893 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 0 33 theme spectrometry-based 27:44 arg1 O-glycomics 46:56 Liquid chromatography mass spectrometry-based O-glycomics 0:56 Liquid chromatography mass spectrometry-based O-glycomics 0:56 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 5 34 theme unpaired 631:638 arg1 test 650:653 unpaired Student t test 631:653 unpaired Student t test 631:653 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 1 35 theme Gastric 123:129 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 35 theme Gastric 123:129 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 6 36 dep O-glycans 818:826 arg1 809.42 845:850 m/z 809.42 841:850 m/z 809.42 841:850 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 36 dep O-glycans 818:826 arg1 m/z 829:831 m/z 733.33 829:838 m/z 733.33 829:838 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 2 37 dep occurs 260:265 arg1 Important 192:200 Important 192:200 Important 192:200 Important for tumorigenesis and progression, aberrant glycosylation occurs frequently in cancers. 26255982 3 38 theme healthy 398:404 arg1 donors 406:411 144 healthy donors 394:411 144 healthy donors 394:411 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 7 39 theme significant 1026:1036 arg1 correlations 1038:1049 significant correlations 1026:1049 significant correlations between the abundance of the O-glycans 1026:1088 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 8 40 theme candidate 1215:1223 arg1 antigens 1225:1232 multiple candidate antigens 1206:1232 multiple candidate antigens for patients with GC 1206:1253 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 4 41 theme O-glycans 465:473 arg1 method 446:451 the method 442:451 the method of labeling O-glycans (released from proteins) 442:498 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 6 42 theme core2 781:785 arg1 level 765:769 The expression level 750:769 The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) 750:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 8 43 theme CONCLUSION 1140:1149 arg1 approaches 1184:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 6 44 dep increased 858:866 arg1 whereas 896:902 whereas 896:902 whereas 896:902 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 44 dep increased 858:866 arg1 < 885:885 p < 0.0001 883:892 p < 0.0001 883:892 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 45 theme antigen 791:797 arg1 level 765:769 The expression level 750:769 The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) 750:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 0 46 theme glycosylation 70:82 arg1 alterations 84:94 glycosylation alterations 70:94 glycosylation alterations in gastric cancer 70:112 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 9 47 theme GC 1310:1311 arg1 serum 1301:1305 the serum 1297:1305 the serum of GC 1297:1311 The O-glycan structures are increased in the serum of GC, they may be candidates for carbohydrate tumor markers. 26255982 0 48 from alterations 84:94 arg1 cancer 107:112 gastric cancer 99:112 gastric cancer 99:112 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 6 49 theme ST 788:789 arg1 antigen 791:797 ST antigen 788:797 ST antigen 788:797 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 8 50 theme CLINICAL 1155:1162 arg1 approaches 1184:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 2 51 theme aberrant 237:244 arg1 glycosylation 246:258 aberrant glycosylation 237:258 aberrant glycosylation 237:258 Important for tumorigenesis and progression, aberrant glycosylation occurs frequently in cancers. 26255982 7 52 gly glycoproteins 1094:1106 arg1 MUC1 1109:1112 MUC1 1109:1112 MUC1 1109:1112 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 7 52 gly glycoproteins 1094:1106 arg1 glycoproteins 1094:1106 glycoproteins 1094:1106 glycoproteins (MUC1, CEA) 1094:1118 In addition, there were significant correlations between the abundance of the O-glycans and glycoproteins (MUC1, CEA) in the serum of GC. 26255982 5 53 theme partial 592:598 arg1 squares 606:612 partial least squares 592:612 partial least squares discriminant 592:625 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 4 54 used used 437:440 arg2 We 434:435 We 434:435 We 434:435 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 5 55 theme positive 727:734 arg1 mode 736:739 positive mode 727:739 positive mode 727:739 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 6 56 theme p 883:883 arg1 < 885:885 p < 0.0001 883:892 p < 0.0001 883:892 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 5 57 theme t 648:648 arg1 test 650:653 unpaired Student t test 631:653 unpaired Student t test 631:653 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 1 58 theme fourth 145:150 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 58 theme fourth 145:150 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 5 59 theme least 600:604 arg1 squares 606:612 partial least squares 592:612 partial least squares discriminant 592:625 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 4 60 theme labeling 456:463 arg1 O-glycans 465:473 labeling O-glycans 456:473 labeling O-glycans (released from proteins) 456:498 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 1 61 theme malignant 164:172 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 61 theme malignant 164:172 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 8 62 theme RELEVANCE 1164:1172 arg1 approaches 1184:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches 1140:1193 CONCLUSION AND CLINICAL RELEVANCE Glycomics approaches identified multiple candidate antigens for patients with GC. 26255982 6 63 theme < 992:992 arg1 p 990:990 p < 0.001 990:998 p < 0.001 990:998 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 6 63 theme < 992:992 arg1 controls 980:987 controls 980:987 controls (p < 0.001) 980:999 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 3 64 theme cancer 369:374 arg1 GC 386:387 GC 386:387 GC 386:387 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 3 64 theme cancer 369:374 arg1 patients 376:383 157 gastric cancer patients 357:383 157 gastric cancer patients (GC) 357:388 We investigated the differences in O-glycosylation in the serum of 157 gastric cancer patients (GC) and 144 healthy donors. 26255982 6 65 theme core1 774:778 arg1 level 765:769 The expression level 750:769 The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) 750:851 RESULTS The expression level of core1, core2, ST antigen, and core2 complex O-glycans (m/z 733.33, m/z 809.42) were increased significantly (p < 0.0001), whereas m/z 529.75 and diST-antigen were decreased in the serum of GC compared with controls (p < 0.001). 26255982 1 66 theme common 157:162 arg1 cancer 131:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer 115:136 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 1 66 theme common 157:162 arg1 worldwide 181:189 the fourth most common malignant cancer worldwide 141:189 the fourth most common malignant cancer worldwide 141:189 PURPOSE Gastric cancer is the fourth most common malignant cancer worldwide. 26255982 5 67 from LC-MS/MS 715:722 arg1 mode 736:739 positive mode 727:739 positive mode 727:739 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 5 67 from LC-MS/MS 715:722 arg1 information 685:695 the structural information 670:695 the structural information of O-glycans from LC-MS/MS in positive mode 670:739 Analyzing the LC-MS data by partial least squares discriminant and unpaired Student t test, combined with the structural information of O-glycans from LC-MS/MS in positive mode. 26255982 2 68 gly glycosylation 246:258 arg2 cancers 281:287 cancers 281:287 cancers 281:287 Important for tumorigenesis and progression, aberrant glycosylation occurs frequently in cancers. 26255982 0 69 theme chromatography 7:20 arg1 O-glycomics 46:56 Liquid chromatography mass spectrometry-based O-glycomics 0:56 Liquid chromatography mass spectrometry-based O-glycomics 0:56 Liquid chromatography mass spectrometry-based O-glycomics to evaluate glycosylation alterations in gastric cancer. 26255982 4 70 dep used 437:440 arg1 followed 536:543 followed 536:543 followed by LC-MS analysis 536:561 EXPERIMENTAL DESIGN We used the method of labeling O-glycans (released from proteins) with 1-phenyl-3-methyl-5-pyrazolone followed by LC-MS analysis. 26255982 9 71 theme carbohydrate 1341:1352 arg1 markers 1360:1366 carbohydrate tumor markers 1341:1366 carbohydrate tumor markers 1341:1366 The O-glycan structures are increased in the serum of GC, they may be candidates for carbohydrate tumor markers. 26248080 6 0 theme fucose 877:882 arg1 structures 915:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 4 1 theme sialylated 646:655 arg1 N-glycans 665:673 sialylated complex N-glycans 646:673 sialylated complex N-glycans 646:673 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 7 2 theme processing 1009:1018 arg1 inhibition 979:988 The inhibition 975:988 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine 975:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 3 3 theme membrane 468:475 arg1 compartments 477:488 cellular membrane compartments 459:488 cellular membrane compartments 459:488 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 7 4 dep glycans 1049:1055 arg1 to 1038:1039 to 1038:1039 to 1038:1039 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 4 5 gly glycoprotein 549:560 arg1 glycoprotein 549:560 glycoprotein galectin-3 binding protein 549:587 The glycoprotein galectin-3 binding protein (LGALS3BP) 545:598 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 3 6 theme compartments 477:488 arg1 GS28 520:523 GS28 520:523 GS28 520:523 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 6 theme compartments 477:488 arg1 LAMP-1 526:531 LAMP-1 526:531 LAMP-1 526:531 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 6 theme compartments 477:488 arg1 markers 448:454 markers 448:454 markers 448:454 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 6 theme compartments 477:488 arg1 calnexin 501:508 calnexin 501:508 calnexin 501:508 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 6 theme compartments 477:488 arg1 GRASP65 511:517 GRASP65 511:517 GRASP65 511:517 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 6 theme compartments 477:488 arg1 L1CAM 538:542 L1CAM 538:542 L1CAM 538:542 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 9 7 theme novel 1405:1409 arg1 biomarkers 1411:1420 novel biomarkers 1405:1420 novel biomarkers for ovarian cancer 1405:1439 Furthermore, the identified glycosignatures of EVs could provide novel biomarkers for ovarian cancer. 26248080 4 8 theme binding 573:579 arg1 LGALS3BP 590:597 LGALS3BP 590:597 LGALS3BP 590:597 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 4 8 theme binding 573:579 arg1 protein 581:587 glycoprotein galectin-3 binding protein 549:587 The glycoprotein galectin-3 binding protein (LGALS3BP) 545:598 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 6 9 with O-glycans 931:939 arg1 T-antigen 950:958 the T-antigen 946:958 the T-antigen 946:958 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 6 9 with O-glycans 931:939 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 8 10 dep composition 1296:1306 arg1 the 1292:1294 the 1292:1294 the 1292:1294 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 2 11 theme cell 295:298 arg1 MBs 311:313 MBs 311:313 MBs 311:313 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 2 11 theme cell 295:298 arg1 membranes 300:308 total cell membranes 289:308 total cell membranes (MBs) 289:314 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 6 12 theme sialic 864:869 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 2 13 theme total 289:293 arg1 MBs 311:313 MBs 311:313 MBs 311:313 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 2 13 theme total 289:293 arg1 membranes 300:308 total cell membranes 289:308 total cell membranes (MBs) 289:314 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 7 14 from glycans 1049:1055 arg1 inhibition 979:988 The inhibition 975:988 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine 975:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 7 14 from glycans 1049:1055 arg1 processing 1009:1018 N-glycosylation processing 993:1018 N-glycosylation processing from high mannose to complex glycans using kifunensine 993:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 6 15 theme α2,3-linked 852:862 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 9 16 theme ovarian 1426:1432 arg1 cancer 1434:1439 ovarian cancer 1426:1439 ovarian cancer 1426:1439 Furthermore, the identified glycosignatures of EVs could provide novel biomarkers for ovarian cancer. 26248080 0 17 from Cells 46:50 arg1 Vesicles 14:21 Extracellular Vesicles 0:21 Extracellular Vesicles from Ovarian Carcinoma Cells 0:50 Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures. 26248080 6 18 with N-glycans 837:845 arg1 T-antigen 950:958 the T-antigen 946:958 the T-antigen 946:958 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 6 18 with N-glycans 837:845 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 4 19 theme glycoprotein 549:560 arg1 LGALS3BP 590:597 LGALS3BP 590:597 LGALS3BP 590:597 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 4 19 theme glycoprotein 549:560 arg1 protein 581:587 glycoprotein galectin-3 binding protein 549:587 The glycoprotein galectin-3 binding protein (LGALS3BP) 545:598 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 3 20 theme specific 372:379 arg1 CD63 408:411 CD63 408:411 CD63 408:411 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 20 theme specific 372:379 arg1 markers 381:387 specific markers 372:387 specific markers 372:387 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 20 theme specific 372:379 arg1 CD9 414:416 CD9 414:416 CD9 414:416 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 20 theme specific 372:379 arg1 Tsg101 400:405 Tsg101 400:405 Tsg101 400:405 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 20 theme specific 372:379 arg1 annexin-I 419:427 annexin-I 419:427 annexin-I 419:427 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 3 20 theme specific 372:379 arg1 MBs 434:436 MBs 434:436 MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM 434:542 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 7 21 from changes 1082:1088 arg1 composition 1097:1107 the composition 1093:1107 the composition of EVs 1093:1114 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 1 22 with environment 130:140 arg1 lipid 185:189 lipid 185:189 lipid 185:189 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 1 22 with environment 130:140 arg1 protein 176:182 protein 176:182 protein 176:182 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 1 22 with environment 130:140 arg1 acid 170:173 characteristic nucleic acid 147:173 characteristic nucleic acid 147:173 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 1 22 with environment 130:140 arg1 composition 203:213 glycan composition 196:213 glycan composition 196:213 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 8 23 theme EVs 1223:1225 arg1 glycosignatures 1204:1218 glycosignatures 1204:1218 glycosignatures of EVs 1204:1225 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 7 24 theme several 1142:1148 arg1 glycoproteins 1150:1162 several glycoproteins 1142:1162 several glycoproteins 1142:1162 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 0 25 theme Extracellular 0:12 arg1 Vesicles 14:21 Extracellular Vesicles 0:21 Extracellular Vesicles from Ovarian Carcinoma Cells 0:50 Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures. 26248080 1 26 theme glycan 196:201 arg1 composition 203:213 glycan composition 196:213 glycan composition 196:213 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 0 27 theme Ovarian 28:34 arg1 Cells 46:50 Ovarian Carcinoma Cells 28:50 Ovarian Carcinoma Cells 28:50 Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures. 26248080 7 28 theme complex 1041:1047 arg1 glycans 1049:1055 high mannose to complex glycans 1025:1055 high mannose to complex glycans using kifunensine 1025:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 5 29 theme specific 736:743 arg1 glycosignatures 745:759 specific glycosignatures 736:759 specific glycosignatures relative to MBs 736:775 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 9 30 theme identified 1357:1366 arg1 glycosignatures 1368:1382 the identified glycosignatures 1353:1382 the identified glycosignatures of EVs 1353:1389 Furthermore, the identified glycosignatures of EVs could provide novel biomarkers for ovarian cancer. 26248080 6 31 theme complex 829:835 arg1 N-glycans 837:845 complex N-glycans 829:845 complex N-glycans with α2,3-linked sialic acid 829:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 1 32 theme extracellular 116:128 arg1 environment 130:140 the extracellular environment 112:140 the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition 112:213 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 8 33 theme release 1331:1337 arg1 composition 1296:1306 composition 1296:1306 composition 1296:1306 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 3 34 theme cellular 459:466 arg1 compartments 477:488 cellular membrane compartments 459:488 cellular membrane compartments 459:488 EVs were enriched in specific markers, including Tsg101, CD63, CD9, annexin-I, and MBs contained markers of cellular membrane compartments, including calnexin, GRASP65, GS28, LAMP-1, and L1CAM. 26248080 7 35 theme mannose 1030:1036 arg1 glycans 1049:1055 high mannose to complex glycans 1025:1055 high mannose to complex glycans using kifunensine 1025:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 7 36 theme glycoproteins 1150:1162 arg1 decrease 1130:1137 a decrease 1128:1137 a decrease of several glycoproteins 1128:1162 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 6 37 gly glycoproteins 807:819 arg1 glycoproteins 807:819 glycoproteins 807:819 glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen 807:958 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 7 38 theme EVs 1112:1114 arg1 composition 1097:1107 the composition 1093:1107 the composition of EVs 1093:1114 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 0 39 theme Carcinoma 36:44 arg1 Cells 46:50 Ovarian Carcinoma Cells 28:50 Ovarian Carcinoma Cells 28:50 Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures. 26248080 5 40 theme Lectin 676:681 arg1 blotting 683:690 Lectin blotting 676:690 Lectin blotting with a panel of lectins 676:714 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 6 41 theme glycoproteins 807:819 arg1 presence 795:802 the presence 791:802 the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen 791:958 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 6 42 link α2,3-linked 852:862 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 0 43 theme Specific 60:67 arg1 Glycosignatures 69:83 Specific Glycosignatures 60:83 Specific Glycosignatures 60:83 Extracellular Vesicles from Ovarian Carcinoma Cells Display Specific Glycosignatures. 26248080 6 44 theme LacdiNAc 906:913 arg1 structures 915:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 5 45 with blotting 683:690 arg1 panel 699:703 a panel 697:703 a panel of lectins 697:714 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 2 46 theme OVMz 339:342 arg1 cells 344:348 ovarian carcinoma OVMz cells 321:348 ovarian carcinoma OVMz cells 321:348 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 7 47 theme high 1025:1028 arg1 glycans 1049:1055 high mannose to complex glycans 1025:1055 high mannose to complex glycans using kifunensine 1025:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 7 48 from inhibition 979:988 arg1 glycans 1049:1055 high mannose to complex glycans 1025:1055 high mannose to complex glycans using kifunensine 1025:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 2 49 theme carcinoma 329:337 arg1 cells 344:348 ovarian carcinoma OVMz cells 321:348 ovarian carcinoma OVMz cells 321:348 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 6 50 theme bisecting-GlcNAc 885:900 arg1 structures 915:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 fucose, bisecting-GlcNAc and LacdiNAc structures 877:924 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 7 51 theme N-glycosylation 993:1007 arg1 processing 1009:1018 N-glycosylation processing 993:1018 N-glycosylation processing from high mannose to complex glycans using kifunensine 993:1073 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 7 52 gly glycoproteins 1150:1162 arg1 glycoproteins 1150:1162 several glycoproteins 1142:1162 several glycoproteins 1142:1162 The inhibition of N-glycosylation processing from high mannose to complex glycans using kifunensine caused changes in the composition of EVs and induced a decrease of several glycoproteins. 26248080 1 53 theme characteristic 147:160 arg1 acid 170:173 characteristic nucleic acid 147:173 characteristic nucleic acid 147:173 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 6 54 with structures 915:924 arg1 T-antigen 950:958 the T-antigen 946:958 the T-antigen 946:958 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 6 54 with structures 915:924 arg1 acid 871:874 α2,3-linked sialic acid 852:874 α2,3-linked sialic acid 852:874 Furthermore, the presence of glycoproteins bearing complex N-glycans with α2,3-linked sialic acid, fucose, bisecting-GlcNAc and LacdiNAc structures, and O-glycans with the T-antigen were detected. 26248080 4 55 theme galectin-3 562:571 arg1 LGALS3BP 590:597 LGALS3BP 590:597 LGALS3BP 590:597 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 4 55 theme galectin-3 562:571 arg1 protein 581:587 glycoprotein galectin-3 binding protein 549:587 The glycoprotein galectin-3 binding protein (LGALS3BP) 545:598 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 4 56 gly sialylated 646:655 arg1 N-glycans 665:673 sialylated complex N-glycans 646:673 sialylated complex N-glycans 646:673 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 5 57 contain had 732:734 arg1 EVs 728:730 EVs 728:730 EVs 728:730 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 5 57 contain had 732:734 arg2 glycosignatures 745:759 specific glycosignatures 736:759 specific glycosignatures relative to MBs 736:775 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 8 58 theme EVs 1327:1329 arg1 release 1331:1337 EVs release 1327:1337 EVs release 1327:1337 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 1 59 theme nucleic 162:168 arg1 acid 170:173 characteristic nucleic acid 147:173 characteristic nucleic acid 147:173 Cells release vesicles to the extracellular environment with characteristic nucleic acid, protein, lipid, and glycan composition. 26248080 2 60 theme ovarian 321:327 arg1 cells 344:348 ovarian carcinoma OVMz cells 321:348 ovarian carcinoma OVMz cells 321:348 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 8 61 gly glycosylation 1253:1265 arg1 release 1331:1337 EVs release 1327:1337 EVs release 1327:1337 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 8 62 theme altered 1245:1251 arg1 glycosylation 1253:1265 altered glycosylation 1245:1265 altered glycosylation within the cell 1245:1281 In conclusion, the results showed that glycosignatures of EVs were specific and altered glycosylation within the cell affected the composition and/or dynamics of EVs release. 26248080 9 63 theme EVs 1387:1389 arg1 glycosignatures 1368:1382 the identified glycosignatures 1353:1382 the identified glycosignatures of EVs 1353:1389 Furthermore, the identified glycosignatures of EVs could provide novel biomarkers for ovarian cancer. 26248080 5 64 theme lectins 708:714 arg1 panel 699:703 a panel 697:703 a panel of lectins 697:714 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 4 65 contain contained 636:644 arg2 N-glycans 665:673 sialylated complex N-glycans 646:673 sialylated complex N-glycans 646:673 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 4 65 contain contained 636:644 arg1 it 633:634 it 633:634 it 633:634 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 26248080 2 66 theme extracellular 256:268 arg1 vesicles 270:277 extracellular vesicles 256:277 extracellular vesicles (EVs) 256:283 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 2 66 theme extracellular 256:268 arg1 EVs 280:282 EVs 280:282 EVs 280:282 Here we have isolated and characterized extracellular vesicles (EVs) and total cell membranes (MBs) from ovarian carcinoma OVMz cells. 26248080 5 67 theme relative 761:768 arg1 glycosignatures 745:759 specific glycosignatures 736:759 specific glycosignatures relative to MBs 736:775 Lectin blotting with a panel of lectins showed that EVs had specific glycosignatures relative to MBs. 26248080 4 68 theme complex 657:663 arg1 N-glycans 665:673 sialylated complex N-glycans 646:673 sialylated complex N-glycans 646:673 The glycoprotein galectin-3 binding protein (LGALS3BP) was strongly enriched in EVs and it contained sialylated complex N-glycans. 28692416 7 0 theme pressure 1518:1525 arg1 status 1527:1532 blood pressure status 1512:1532 blood pressure status in the Kazakh population 1512:1557 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 6 1 theme status 1146:1151 arg1 differentiation 1102:1116 differentiation 1102:1116 differentiation of hypertension and healthy status in the Kazakh population sample 1102:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 2 2 theme immunoglobulin 364:377 arg1 G 379:379 immunoglobulin G 364:379 immunoglobulin G (IgG) glycome composition 364:405 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 2 2 theme immunoglobulin 364:377 arg1 IgG 382:384 IgG 382:384 IgG 382:384 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 5 3 gly N-glycopeptide 911:924 arg2 N-glycopeptide 911:924 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 3 4 from hypertension 641:652 arg1 population 668:677 the Kazakh population 657:677 the Kazakh population 657:677 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 1 5 theme clinical 277:284 arg1 medicine 286:293 clinical medicine 277:293 clinical medicine 277:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 7 6 gly N-glycopeptide 1444:1457 arg2 N-glycopeptide 1444:1457 Fc N-glycopeptide profiles 1441:1466 Fc N-glycopeptide profiles of plasma IgG subclasses 1441:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 9 7 theme Further 1712:1718 arg1 studies 1720:1726 Further studies 1712:1726 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases 1712:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 7 8 theme Fc 1441:1442 arg1 profiles 1459:1466 Fc N-glycopeptide profiles 1441:1466 Fc N-glycopeptide profiles of plasma IgG subclasses 1441:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 3 9 theme first 554:558 arg1 time 560:563 the first time 550:563 the first time to the best of our knowledge 550:592 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 5 10 theme subclass-specific 890:906 arg1 structures 926:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 7 11 theme IgG 1478:1480 arg1 subclasses 1482:1491 plasma IgG subclasses 1471:1491 plasma IgG subclasses 1471:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 6 12 theme characteristic 1209:1222 arg1 analysis 1230:1237 characteristic curve analysis 1209:1237 characteristic curve analysis 1209:1237 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 6 13 theme IgG 1296:1298 arg1 N-glycans 1300:1308 nine IgG N-glycans 1291:1308 nine IgG N-glycans 1291:1308 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 1 14 theme fresh 193:197 arg1 vistas 199:204 fresh vistas 193:204 fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine 193:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 9 15 theme glycomic 1743:1750 arg1 biomarkers 1752:1761 postgenomic glycomic biomarkers 1731:1761 postgenomic glycomic biomarkers in cardiovascular and chronic diseases 1731:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 6 16 theme mass 1366:1369 arg1 index 1371:1375 body mass index 1361:1375 body mass index 1361:1375 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 9 17 from studies 1720:1726 arg1 diseases 1793:1800 cardiovascular and chronic diseases 1766:1800 cardiovascular and chronic diseases 1766:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 1 18 from diagnostics 262:272 arg1 medicine 286:293 clinical medicine 277:293 clinical medicine 277:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 3 19 theme IgG 599:601 arg1 N-glycans 603:611 the IgG N-glycans 595:611 the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population 595:677 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 1 20 from innovation 210:219 arg1 biomarkers 240:249 chronic disease biomarkers 224:249 chronic disease biomarkers 224:249 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 1 20 from innovation 210:219 arg1 diagnostics 262:272 system diagnostics 255:272 system diagnostics in clinical medicine 255:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 0 21 from Hypertension 67:78 arg1 Population 94:103 the Kazakh Population 83:103 the Kazakh Population 83:103 Immunoglobulin G N-Glycans as Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population. 28692416 4 22 theme IgG 717:719 arg1 subclass 721:728 IgG subclass 717:728 IgG subclass isolated from plasma samples of 150 Kazakh study participants 717:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 5 23 theme blood 1082:1086 arg1 pressure 1088:1095 diastolic blood pressure 1072:1095 diastolic blood pressure 1072:1095 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 5 24 theme derived 953:959 arg1 trait 975:979 one derived glycosylation trait 949:979 one derived glycosylation trait in subclasses IgG2/3 and IgG4 949:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 4 25 gly N-glycopeptides 698:712 arg1 subclass 721:728 IgG subclass 717:728 IgG subclass isolated from plasma samples of 150 Kazakh study participants 717:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 4 25 gly N-glycopeptides 698:712 arg2 N-glycopeptides 698:712 60 N-glycopeptides 695:712 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants 695:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 7 26 from alteration 1427:1436 arg1 profiles 1459:1466 Fc N-glycopeptide profiles 1441:1466 Fc N-glycopeptide profiles of plasma IgG subclasses 1441:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 1 27 theme nascent 156:162 arg1 field 164:168 the nascent field 152:168 the nascent field of glycomics 152:181 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 4 28 theme N-glycopeptides 698:712 arg1 profile 684:690 The profile 680:690 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants 680:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 1 29 from field 164:168 arg1 biomarkers 136:145 Next-generation (postgenomic) biomarkers 106:145 Next-generation (postgenomic) biomarkers from the nascent field of glycomics 106:181 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 6 30 theme curve 1224:1228 arg1 analysis 1230:1237 characteristic curve analysis 1209:1237 characteristic curve analysis 1209:1237 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 4 31 theme Kazakh 766:771 arg1 participants 779:790 150 Kazakh study participants 762:790 150 Kazakh study participants 762:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 9 32 from biomarkers 1752:1761 arg1 diseases 1793:1800 cardiovascular and chronic diseases 1766:1800 cardiovascular and chronic diseases 1766:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 4 33 theme participants 779:790 arg1 samples 751:757 plasma samples 744:757 plasma samples of 150 Kazakh study participants 744:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 6 34 theme hypertension 1121:1132 arg1 differentiation 1102:1116 differentiation 1102:1116 differentiation of hypertension and healthy status in the Kazakh population sample 1102:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 2 35 theme IgG 467:469 arg1 N-glycosylation 448:462 N-glycosylation 448:462 N-glycosylation of IgG 448:469 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 8 36 theme IgG 1560:1562 arg1 biomarkers 1612:1621 potential biomarkers 1602:1621 potential biomarkers for hypertension in the Kazakhs 1602:1653 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 8 36 theme IgG 1560:1562 arg1 profiles 1580:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 4 37 theme plasma 744:749 arg1 samples 751:757 plasma samples 744:757 plasma samples of 150 Kazakh study participants 744:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 2 38 theme glycome 387:393 arg1 composition 395:405 immunoglobulin G (IgG) glycome composition 364:405 immunoglobulin G (IgG) glycome composition 364:405 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 0 39 theme Immunoglobulin 0:13 arg1 G 15:15 Immunoglobulin G 0:15 Immunoglobulin G 0:15 Immunoglobulin G N-Glycans as Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population. 28692416 2 40 theme hypertension 491:502 arg1 pathogenesis 504:515 hypertension pathogenesis 491:515 hypertension pathogenesis 491:515 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 4 41 theme liquid 831:836 arg1 chromatography 838:851 nano ultra-performance liquid chromatography 808:851 nano ultra-performance liquid chromatography with mass spectrometry 808:874 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 5 42 theme IgG 886:888 arg1 structures 926:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 0 43 theme Potential 30:38 arg1 Biomarkers 52:61 Potential Postgenomic Biomarkers 30:61 Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population 30:103 Immunoglobulin G N-Glycans as Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population. 28692416 8 44 theme potential 1602:1610 arg1 biomarkers 1612:1621 potential biomarkers 1602:1621 potential biomarkers for hypertension in the Kazakhs 1602:1653 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 8 44 theme potential 1602:1610 arg1 profiles 1580:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 5 45 theme diastolic 1072:1080 arg1 pressure 1088:1095 diastolic blood pressure 1072:1095 diastolic blood pressure 1072:1095 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 6 46 from sample 1178:1183 arg1 differentiation 1102:1116 differentiation 1102:1116 differentiation of hypertension and healthy status in the Kazakh population sample 1102:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 1 47 theme chronic 224:230 arg1 biomarkers 240:249 chronic disease biomarkers 224:249 chronic disease biomarkers 224:249 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 5 48 link derived 953:959 arg1 trait 975:979 one derived glycosylation trait 949:979 one derived glycosylation trait in subclasses IgG2/3 and IgG4 949:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 6 49 theme Kazakh 1160:1165 arg1 sample 1178:1183 the Kazakh population sample 1156:1183 the Kazakh population sample 1156:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 4 50 theme nano 808:811 arg1 chromatography 838:851 nano ultra-performance liquid chromatography 808:851 nano ultra-performance liquid chromatography with mass spectrometry 808:874 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 7 51 theme blood 1512:1516 arg1 status 1527:1532 blood pressure status 1512:1532 blood pressure status in the Kazakh population 1512:1557 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 5 52 theme systolic 1041:1048 arg1 pressure 1056:1063 systolic blood pressure 1041:1063 systolic blood pressure 1041:1063 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 2 53 from variation 435:443 arg1 N-glycosylation 448:462 N-glycosylation 448:462 N-glycosylation of IgG 448:469 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 6 54 theme traditional 1332:1342 arg1 gender 1344:1349 the traditional gender 1328:1349 the traditional gender 1328:1349 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 2 55 theme G 379:379 arg1 composition 395:405 immunoglobulin G (IgG) glycome composition 364:405 immunoglobulin G (IgG) glycome composition 364:405 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 1 56 theme system 255:260 arg1 diagnostics 262:272 system diagnostics 255:272 system diagnostics in clinical medicine 255:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 6 57 theme healthy 1138:1144 arg1 status 1146:1151 healthy status 1138:1151 healthy status 1138:1151 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 4 58 attach isolated 730:737 arg1 samples 751:757 plasma samples 744:757 plasma samples of 150 Kazakh study participants 744:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 4 58 attach isolated 730:737 arg2 subclass 721:728 IgG subclass 717:728 IgG subclass isolated from plasma samples of 150 Kazakh study participants 717:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 6 59 from differentiation 1102:1116 arg1 sample 1178:1183 the Kazakh population sample 1156:1183 the Kazakh population sample 1156:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 2 60 theme previous 300:307 arg1 work 309:312 Our previous work 296:312 Our previous work 296:312 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 7 61 theme subclasses 1482:1491 arg1 profiles 1459:1466 Fc N-glycopeptide profiles 1441:1466 Fc N-glycopeptide profiles of plasma IgG subclasses 1441:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 8 62 from hypertension 1627:1638 arg1 Kazakhs 1647:1653 the Kazakhs 1643:1653 the Kazakhs 1643:1653 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 6 63 from hypertension 1121:1132 arg1 sample 1178:1183 the Kazakh population sample 1156:1183 the Kazakh population sample 1156:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 1 64 theme glycomics 173:181 arg1 field 164:168 the nascent field 152:168 the nascent field of glycomics 152:181 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 7 65 theme plasma 1471:1476 arg1 subclasses 1482:1491 plasma IgG subclasses 1471:1491 plasma IgG subclasses 1471:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 4 66 theme mass 858:861 arg1 spectrometry 863:874 mass spectrometry 858:874 mass spectrometry 858:874 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 7 67 theme N-glycopeptide 1444:1457 arg1 profiles 1459:1466 Fc N-glycopeptide profiles 1441:1466 Fc N-glycopeptide profiles of plasma IgG subclasses 1441:1491 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 5 68 theme Fc 908:909 arg1 structures 926:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 9 69 theme postgenomic 1731:1741 arg1 biomarkers 1752:1761 postgenomic glycomic biomarkers 1731:1761 postgenomic glycomic biomarkers in cardiovascular and chronic diseases 1731:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 5 70 dep along 938:942 arg1 with 944:947 with 944:947 with 944:947 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 9 71 theme biomarkers 1752:1761 arg1 studies 1720:1726 Further studies 1712:1726 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases 1712:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 6 72 theme body 1361:1364 arg1 index 1371:1375 body mass index 1361:1375 body mass index 1361:1375 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 5 73 theme N-glycopeptide 911:924 arg1 structures 926:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures 877:935 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 9 74 theme cardiovascular 1766:1779 arg1 diseases 1793:1800 cardiovascular and chronic diseases 1766:1800 cardiovascular and chronic diseases 1766:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 1 75 from biomarkers 240:249 arg1 medicine 286:293 clinical medicine 277:293 clinical medicine 277:293 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 1 76 theme postgenomic 123:133 arg1 biomarkers 136:145 Next-generation (postgenomic) biomarkers 106:145 Next-generation (postgenomic) biomarkers from the nascent field of glycomics 106:181 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 9 77 theme chronic 1785:1791 arg1 diseases 1793:1800 cardiovascular and chronic diseases 1766:1800 cardiovascular and chronic diseases 1766:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 6 78 theme model 1274:1278 arg1 performance 1255:1265 the performance 1251:1265 the performance 1251:1265 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 6 78 theme model 1274:1278 arg1 greater 1315:1321 greater 1315:1321 greater 1315:1321 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 3 79 theme potential 616:624 arg1 biomarkers 626:635 potential biomarkers 616:635 potential biomarkers for hypertension in the Kazakh population 616:677 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 5 80 theme glycosylation 961:973 arg1 trait 975:979 one derived glycosylation trait 949:979 one derived glycosylation trait in subclasses IgG2/3 and IgG4 949:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 9 81 from diseases 1793:1800 arg1 studies 1720:1726 Further studies 1712:1726 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases 1712:1800 Further studies of postgenomic glycomic biomarkers in cardiovascular and chronic diseases are timely and called for. 28692416 4 82 theme subclass 721:728 arg1 N-glycopeptides 698:712 60 N-glycopeptides 695:712 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants 695:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 4 83 with chromatography 838:851 arg1 spectrometry 863:874 mass spectrometry 858:874 mass spectrometry 858:874 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 0 84 theme Kazakh 87:92 arg1 Population 94:103 the Kazakh Population 83:103 the Kazakh Population 83:103 Immunoglobulin G N-Glycans as Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population. 28692416 5 85 dep subclasses 984:993 arg1 IgG2/3 995:1000 IgG2/3 995:1000 IgG2/3 995:1000 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 5 85 dep subclasses 984:993 arg1 subclasses 984:993 subclasses IgG2/3 and IgG4 984:1009 subclasses IgG2/3 and IgG4 984:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 5 85 dep subclasses 984:993 arg1 IgG4 1006:1009 IgG4 1006:1009 IgG4 1006:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 8 86 theme N-glycosylation 1564:1578 arg1 biomarkers 1612:1621 potential biomarkers 1602:1621 potential biomarkers for hypertension in the Kazakhs 1602:1653 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 8 86 theme N-glycosylation 1564:1578 arg1 profiles 1580:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles 1560:1587 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 4 87 theme study 773:777 arg1 participants 779:790 150 Kazakh study participants 762:790 150 Kazakh study participants 762:790 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 5 88 from trait 975:979 arg1 IgG2/3 995:1000 IgG2/3 995:1000 IgG2/3 995:1000 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 5 88 from trait 975:979 arg1 subclasses 984:993 subclasses IgG2/3 and IgG4 984:1009 subclasses IgG2/3 and IgG4 984:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 5 88 from trait 975:979 arg1 IgG4 1006:1009 IgG4 1006:1009 IgG4 1006:1009 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 1 89 theme Next-generation 106:120 arg1 biomarkers 136:145 Next-generation (postgenomic) biomarkers 106:145 Next-generation (postgenomic) biomarkers from the nascent field of glycomics 106:181 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 2 90 theme individual 424:433 arg1 variation 435:443 individual variation 424:443 individual variation in N-glycosylation of IgG 424:469 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 0 91 theme Postgenomic 40:50 arg1 Biomarkers 52:61 Potential Postgenomic Biomarkers 30:61 Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population 30:103 Immunoglobulin G N-Glycans as Potential Postgenomic Biomarkers for Hypertension in the Kazakh Population. 28692416 5 92 theme blood 1050:1054 arg1 pressure 1056:1063 systolic blood pressure 1041:1063 systolic blood pressure 1041:1063 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 4 93 theme ultra-performance 813:829 arg1 chromatography 838:851 nano ultra-performance liquid chromatography 808:851 nano ultra-performance liquid chromatography with mass spectrometry 808:874 The profile of 60 N-glycopeptides of IgG subclass isolated from plasma samples of 150 Kazakh study participants was analyzed by nano ultra-performance liquid chromatography with mass spectrometry. 28692416 3 94 theme present 522:528 arg1 study 530:534 The present study 518:534 The present study 518:534 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 8 95 theme personalized 1689:1700 arg1 medicine 1702:1709 personalized medicine 1689:1709 personalized medicine 1689:1709 IgG N-glycosylation profiles may serve as potential biomarkers for hypertension in the Kazakhs, thus contributing to move toward personalized medicine. 28692416 2 96 gly N-glycosylation 448:462 arg1 IgG 467:469 IgG 467:469 IgG 467:469 Our previous work has shown an association between hypertension and immunoglobulin G (IgG) glycome composition, suggesting that individual variation in N-glycosylation of IgG might contribute to hypertension pathogenesis. 28692416 7 97 theme Kazakh 1541:1546 arg1 population 1548:1557 the Kazakh population 1537:1557 the Kazakh population 1537:1557 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 28692416 6 98 theme population 1167:1176 arg1 sample 1178:1183 the Kazakh population sample 1156:1183 the Kazakh population sample 1156:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 5 99 dep derived 953:959 arg1 one 949:951 one 949:951 one 949:951 Fourteen IgG subclass-specific Fc N-glycopeptide structures, along with one derived glycosylation trait in subclasses IgG2/3 and IgG4, were found to correlate with systolic blood pressure and/or diastolic blood pressure. 28692416 3 100 theme Kazakh 661:666 arg1 population 668:677 the Kazakh population 657:677 the Kazakh population 657:677 The present study examined, for the first time to the best of our knowledge, the IgG N-glycans as potential biomarkers for hypertension in the Kazakh population. 28692416 6 101 from status 1146:1151 arg1 sample 1178:1183 the Kazakh population sample 1156:1183 the Kazakh population sample 1156:1183 For differentiation of hypertension and healthy status in the Kazakh population sample, the receiver operating characteristic curve analysis showed that the performance of the model, including nine IgG N-glycans, was greater than the traditional gender, age, and body mass index based model (p < 0.05). 28692416 1 102 theme disease 232:238 arg1 biomarkers 240:249 chronic disease biomarkers 224:249 chronic disease biomarkers 224:249 Next-generation (postgenomic) biomarkers from the nascent field of glycomics now offer fresh vistas for innovation in chronic disease biomarkers and system diagnostics in clinical medicine. 28692416 7 103 from status 1527:1532 arg1 population 1548:1557 the Kazakh population 1537:1557 the Kazakh population 1537:1557 This study indicates that alteration in Fc N-glycopeptide profiles of plasma IgG subclasses is associated with blood pressure status in the Kazakh population. 25182382 9 0 gly N-glycopeptides 1693:1707 arg2 N-glycopeptides 1693:1707 intact N-glycopeptides 1686:1707 intact N-glycopeptides derived from complex proteome samples 1686:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 5 1 from enrichment 1042:1051 arg1 peptides 1013:1020 deglycosylated peptides 998:1020 deglycosylated peptides from the same HILIC enrichment 998:1051 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 1 2 gly glycopeptides 291:303 arg2 glycopeptides 291:303 intact glycopeptides 284:303 intact glycopeptides 284:303 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 3 3 gly glycopeptides 570:582 arg2 glycopeptides 570:582 intact glycopeptides 563:582 intact glycopeptides as well as de-glycopeptides 563:610 N-glycopeptides were identified using the spectra acquired for intact glycopeptides as well as de-glycopeptides. 25182382 4 4 theme glycan 730:735 arg1 structure 717:725 the structure 713:725 the structure of glycan 713:735 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 1 5 theme attachment 221:230 arg1 sites 232:236 specific attachment sites 212:236 specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry 212:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 9 6 theme BIOLOGICAL 1569:1578 arg1 SIGNIFICANCE 1580:1591 BIOLOGICAL SIGNIFICANCE 1569:1591 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 1569:1746 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 8 7 theme glycosylation 1488:1500 arg1 sites 1502:1506 453 glycosylation sites 1484:1506 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations 1484:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 9 8 theme software 1624:1631 arg1 platform 1633:1640 a fully automated software platform 1606:1640 a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1606:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 5 9 theme intact 894:899 arg1 glycopeptides 901:913 intact glycopeptides 894:913 intact glycopeptides 894:913 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 4 10 theme glycan 784:789 arg1 database 791:798 a constructed glycan database 770:798 a constructed glycan database with calculated molecular weight of glycans and their fragment ions 770:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 1 11 from proteins 241:248 arg1 identification 171:184 the identification 167:184 the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry 167:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 9 12 theme high 1650:1653 arg1 characterization 1666:1681 the high throughput characterization 1646:1681 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1646:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 9 13 dep SIGNIFICANCE 1580:1591 arg1 presented 1596:1604 presented 1596:1604 presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1596:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 1 14 from sites 232:236 arg1 proteins 241:248 proteins 241:248 proteins 241:248 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 9 15 theme throughput 1655:1664 arg1 characterization 1666:1681 the high throughput characterization 1646:1681 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1646:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 13 16 theme intact 2128:2133 arg1 glycopeptides 2135:2147 2249 intact glycopeptides 2123:2147 2249 intact glycopeptides 2123:2147 In total, 2249 intact glycopeptides were identified which is by the far the largest dataset among the studies of N-glycoproteomics. 25182382 11 17 theme times 1950:1954 arg1 correlation 1925:1935 The strong correlation 1914:1935 The strong correlation of retention times 1914:1954 The strong correlation of retention times effectively filtered out random matches. 25182382 0 18 theme glycoproteomic 74:87 arg1 method 89:94 an automated glycoproteomic method 61:94 an automated glycoproteomic method 61:94 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 5 19 theme Y1 980:981 arg1 ion 983:985 Y1 ion 980:985 Y1 ion 980:985 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 7 20 theme 293T 1333:1336 arg1 cells 1338:1342 HEK 293T cells 1329:1342 HEK 293T cells 1329:1342 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 1 21 from composition 196:206 arg1 proteins 241:248 proteins 241:248 proteins 241:248 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 3 22 gly de-glycopeptides 595:610 arg2 de-glycopeptides 595:610 de-glycopeptides 595:610 intact glycopeptides as well as de-glycopeptides 563:610 N-glycopeptides were identified using the spectra acquired for intact glycopeptides as well as de-glycopeptides. 25182382 1 23 theme site-specific 130:142 arg1 glycosylation 144:156 site-specific glycosylation 130:156 site-specific glycosylation 130:156 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 11 24 theme random 1981:1986 arg1 matches 1988:1994 random matches 1981:1994 random matches 1981:1994 The strong correlation of retention times effectively filtered out random matches. 25182382 9 25 theme complex 1722:1728 arg1 samples 1739:1745 complex proteome samples 1722:1745 complex proteome samples 1722:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 10 26 theme weights 1884:1890 arg1 commonality 1859:1869 the commonality 1855:1869 the commonality of molecular weights of peptide backbones 1855:1911 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 4 27 theme fragment 854:861 arg1 ions 863:866 their fragment ions 848:866 their fragment ions 848:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 2 28 theme N-glycopeptides 445:459 arg1 characterization 418:433 the high throughput characterization 398:433 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 398:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 7 29 theme site-specific 1298:1310 arg1 glycosylation 1312:1324 site-specific glycosylation 1298:1324 site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides 1298:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 6 30 theme above 1162:1166 arg1 processes 1168:1176 the above processes 1158:1176 the above processes involved in the identification of the intact N-glycopeptides 1158:1237 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 7 31 from characterization 1278:1293 arg1 cells 1338:1342 HEK 293T cells 1329:1342 HEK 293T cells 1329:1342 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 4 32 theme molecular 816:824 arg1 weight 826:831 calculated molecular weight 805:831 calculated molecular weight of glycans and their fragment ions 805:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 0 33 theme Large-scale 0:10 arg1 characterization 12:27 Large-scale characterization 0:27 Large-scale characterization of intact N-glycopeptides 0:53 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 9 34 theme intact 1686:1691 arg1 N-glycopeptides 1693:1707 intact N-glycopeptides 1686:1707 intact N-glycopeptides derived from complex proteome samples 1686:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 10 35 gly deglycosylated 1779:1792 arg1 forms 1794:1798 their deglycosylated forms 1773:1798 their deglycosylated forms 1773:1798 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 2 36 theme proteome 482:489 arg1 samples 491:497 complex proteome samples 474:497 complex proteome samples 474:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 5 37 theme glycopeptides 901:913 arg1 sequences 881:889 The peptide sequences 869:889 The peptide sequences of intact glycopeptides 869:913 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 12 38 theme random 2084:2089 arg1 matches 2091:2097 random matches 2084:2097 random matches 2084:2097 The reliability of this strategy was carefully evaluated which showed a probability of random matches less than 1%. 25182382 5 39 theme HILIC 1036:1040 arg1 enrichment 1042:1051 the same HILIC enrichment 1027:1051 the same HILIC enrichment 1027:1051 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 8 40 theme high 1531:1534 arg1 heterogeneity 1536:1548 the high heterogeneity 1527:1548 the high heterogeneity of glycosylations 1527:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 10 41 theme molecular 1874:1882 arg1 weights 1884:1890 molecular weights 1874:1890 molecular weights of peptide backbones 1874:1911 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 0 42 theme N-glycopeptides 39:53 arg1 characterization 12:27 Large-scale characterization 0:27 Large-scale characterization of intact N-glycopeptides 0:53 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 8 43 gly glycosylation 1488:1500 arg2 sites 1502:1506 453 glycosylation sites 1484:1506 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations 1484:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 8 43 gly glycosylation 1488:1500 arg2 453 1484:1486 453 1484:1486 453 1484:1486 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 2 44 attach derived 461:467 arg1 samples 491:497 complex proteome samples 474:497 complex proteome samples 474:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 2 44 attach derived 461:467 arg2 N-glycopeptides 445:459 intact N-glycopeptides 438:459 intact N-glycopeptides derived from complex proteome samples 438:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 7 45 theme N-glycopeptides 1399:1413 arg1 identification 1362:1375 the identification 1358:1375 the identification of 2249 unique intact N-glycopeptides 1358:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 5 46 gly glycopeptides 901:913 arg2 glycopeptides 901:913 intact glycopeptides 894:913 intact glycopeptides 894:913 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 3 47 theme intact 563:568 arg1 glycopeptides 570:582 intact glycopeptides 563:582 intact glycopeptides as well as de-glycopeptides 563:610 N-glycopeptides were identified using the spectra acquired for intact glycopeptides as well as de-glycopeptides. 25182382 5 48 theme peptide 873:879 arg1 sequences 881:889 The peptide sequences 869:889 The peptide sequences of intact glycopeptides 869:913 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 6 49 theme intact 1216:1221 arg1 N-glycopeptides 1223:1237 the intact N-glycopeptides 1212:1237 the intact N-glycopeptides 1212:1237 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 4 50 theme Y1 617:618 arg1 peptide+GlcNAc 625:638 peptide+GlcNAc 625:638 peptide+GlcNAc 625:638 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 4 50 theme Y1 617:618 arg1 ion 620:622 The Y1 ion 613:622 The Y1 ion (peptide+GlcNAc) 613:639 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 9 51 attach derived 1709:1715 arg1 samples 1739:1745 complex proteome samples 1722:1745 complex proteome samples 1722:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 9 51 attach derived 1709:1715 arg2 N-glycopeptides 1693:1707 intact N-glycopeptides 1686:1707 intact N-glycopeptides derived from complex proteome samples 1686:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 11 52 theme retention 1940:1948 arg1 times 1950:1954 retention times 1940:1954 retention times 1940:1954 The strong correlation of retention times effectively filtered out random matches. 25182382 13 53 theme N-glycoproteomics 2226:2242 arg1 studies 2215:2221 the studies 2211:2221 the studies of N-glycoproteomics 2211:2242 In total, 2249 intact glycopeptides were identified which is by the far the largest dataset among the studies of N-glycoproteomics. 25182382 10 54 theme backbones 1903:1911 arg1 weights 1884:1890 molecular weights 1874:1890 molecular weights of peptide backbones 1874:1911 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 5 55 theme separated 1073:1081 arg1 analysis 1092:1099 a separated LC-MS/MS analysis 1071:1099 a separated LC-MS/MS analysis 1071:1099 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 1 56 theme glycan 189:194 arg1 composition 196:206 glycan composition 189:206 glycan composition 189:206 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 6 57 theme software 1122:1129 arg1 platform 1131:1138 The fully automated software platform 1102:1138 The fully automated software platform 1102:1138 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 13 58 theme largest 2189:2195 arg1 dataset 2197:2203 the largest dataset 2185:2203 the far the largest dataset 2177:2203 In total, 2249 intact glycopeptides were identified which is by the far the largest dataset among the studies of N-glycoproteomics. 25182382 2 59 theme throughput 407:416 arg1 characterization 418:433 the high throughput characterization 398:433 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 398:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 5 60 theme molecular 947:955 arg1 weight 957:962 the molecular weight 943:962 the molecular weight calculated from Y1 ion 943:985 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 8 61 gly heterogeneity 1536:1548 arg1 glycosylations 1553:1566 glycosylations 1553:1566 glycosylations 1553:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 12 62 theme strategy 2021:2028 arg1 reliability 2001:2011 The reliability 1997:2011 The reliability of this strategy 1997:2028 The reliability of this strategy was carefully evaluated which showed a probability of random matches less than 1%. 25182382 4 63 theme intact 687:692 arg1 glycopeptides 694:706 intact glycopeptides 687:706 intact glycopeptides 687:706 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 4 64 with database 791:798 arg1 weight 826:831 calculated molecular weight 805:831 calculated molecular weight of glycans and their fragment ions 805:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 1 65 theme glycopeptides 291:303 arg1 identification 266:279 the identification 262:279 the identification of intact glycopeptides by mass spectrometry 262:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 1 66 theme mass 308:311 arg1 spectrometry 313:324 mass spectrometry 308:324 mass spectrometry 308:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 9 67 theme N-glycopeptides 1693:1707 arg1 characterization 1666:1681 the high throughput characterization 1646:1681 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1646:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 10 68 theme Intact 1748:1753 arg1 glycopeptides 1755:1767 Intact glycopeptides 1748:1767 Intact glycopeptides 1748:1767 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 1 69 theme specific 212:219 arg1 sites 232:236 specific attachment sites 212:236 specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry 212:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 8 70 theme glycosylations 1553:1566 arg1 heterogeneity 1536:1548 the high heterogeneity 1527:1548 the high heterogeneity of glycosylations 1527:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 1 71 theme sites 232:236 arg1 identification 171:184 the identification 167:184 the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry 167:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 2 72 attach present 345:351 arg2 we 342:343 we 342:343 we 342:343 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 2 72 attach present 345:351 arg1 study 335:339 this study 330:339 this study 330:339 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 10 73 theme deglycosylated 1779:1792 arg1 forms 1794:1798 their deglycosylated forms 1773:1798 their deglycosylated forms 1773:1798 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 9 74 theme automated 1614:1622 arg1 platform 1633:1640 a fully automated software platform 1606:1640 a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 1606:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 7 75 theme unique 1385:1390 arg1 N-glycopeptides 1399:1413 2249 unique intact N-glycopeptides 1380:1413 2249 unique intact N-glycopeptides 1380:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 13 76 gly glycopeptides 2135:2147 arg2 glycopeptides 2135:2147 2249 intact glycopeptides 2123:2147 2249 intact glycopeptides 2123:2147 In total, 2249 intact glycopeptides were identified which is by the far the largest dataset among the studies of N-glycoproteomics. 25182382 5 77 gly deglycosylated 998:1011 arg1 peptides 1013:1020 deglycosylated peptides 998:1020 deglycosylated peptides from the same HILIC enrichment 998:1051 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 7 78 from glycosylation 1312:1324 arg1 cells 1338:1342 HEK 293T cells 1329:1342 HEK 293T cells 1329:1342 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 12 79 dep % 2110:2110 arg1 1 2109:2109 1 2109:2109 1 2109:2109 The reliability of this strategy was carefully evaluated which showed a probability of random matches less than 1%. 25182382 10 80 gly glycopeptides 1755:1767 arg2 glycopeptides 1755:1767 Intact glycopeptides 1748:1767 Intact glycopeptides 1748:1767 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 1 81 theme glycosylation 144:156 arg1 characterization 110:125 The detailed characterization 97:125 The detailed characterization of site-specific glycosylation 97:156 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 4 82 theme constructed 772:782 arg1 database 791:798 a constructed glycan database 770:798 a constructed glycan database with calculated molecular weight of glycans and their fragment ions 770:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 5 83 theme deglycosylated 998:1011 arg1 peptides 1013:1020 deglycosylated peptides 998:1020 deglycosylated peptides from the same HILIC enrichment 998:1051 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 6 84 theme automated 1112:1120 arg1 platform 1131:1138 The fully automated software platform 1102:1138 The fully automated software platform 1102:1138 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 7 85 from cells 1338:1342 arg1 characterization 1278:1293 detailed characterization 1269:1293 detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides 1269:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 13 86 theme 2249 2123:2126 arg1 glycopeptides 2135:2147 2249 intact glycopeptides 2123:2147 2249 intact glycopeptides 2123:2147 In total, 2249 intact glycopeptides were identified which is by the far the largest dataset among the studies of N-glycoproteomics. 25182382 8 87 theme intact 1422:1427 arg1 glycopeptides 1429:1441 These intact glycopeptides 1416:1441 These intact glycopeptides 1416:1441 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 7 88 theme HEK 1329:1331 arg1 cells 1338:1342 HEK 293T cells 1329:1342 HEK 293T cells 1329:1342 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 1 89 theme composition 196:206 arg1 identification 171:184 the identification 167:184 the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry 167:324 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 7 90 theme glycosylation 1312:1324 arg1 characterization 1278:1293 detailed characterization 1269:1293 detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides 1269:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 3 91 gly N-glycopeptides 500:514 arg2 N-glycopeptides 500:514 N-glycopeptides 500:514 N-glycopeptides 500:514 N-glycopeptides were identified using the spectra acquired for intact glycopeptides as well as de-glycopeptides. 25182382 7 92 gly N-glycopeptides 1399:1413 arg2 N-glycopeptides 1399:1413 2249 unique intact N-glycopeptides 1380:1413 2249 unique intact N-glycopeptides 1380:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 4 93 theme glycans 836:842 arg1 weight 826:831 calculated molecular weight 805:831 calculated molecular weight of glycans and their fragment ions 805:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 7 94 gly glycosylation 1312:1324 arg1 cells 1338:1342 HEK 293T cells 1329:1342 HEK 293T cells 1329:1342 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 10 95 theme peptide 1895:1901 arg1 backbones 1903:1911 peptide backbones 1895:1911 peptide backbones 1895:1911 Intact glycopeptides and their deglycosylated forms were identified respectively and combined according to the commonality of molecular weights of peptide backbones. 25182382 8 96 gly glycopeptides 1429:1441 arg2 glycopeptides 1429:1441 These intact glycopeptides 1416:1441 These intact glycopeptides 1416:1441 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 4 97 gly glycopeptides 694:706 arg2 glycopeptides 694:706 intact glycopeptides 687:706 intact glycopeptides 687:706 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 9 98 theme proteome 1730:1737 arg1 samples 1739:1745 complex proteome samples 1722:1745 complex proteome samples 1722:1745 BIOLOGICAL SIGNIFICANCE We presented a fully automated software platform for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 8 99 from N-glycans 1471:1479 arg1 sites 1502:1506 453 glycosylation sites 1484:1506 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations 1484:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 2 100 gly N-glycopeptides 445:459 arg2 N-glycopeptides 445:459 intact N-glycopeptides 438:459 intact N-glycopeptides derived from complex proteome samples 438:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 2 101 theme intact 438:443 arg1 N-glycopeptides 445:459 intact N-glycopeptides 438:459 intact N-glycopeptides derived from complex proteome samples 438:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 7 102 theme detailed 1269:1276 arg1 characterization 1278:1293 detailed characterization 1269:1293 detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides 1269:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 1 103 from identification 171:184 arg1 proteins 241:248 proteins 241:248 proteins 241:248 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 1 104 theme detailed 101:108 arg1 characterization 110:125 The detailed characterization 97:125 The detailed characterization of site-specific glycosylation 97:156 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25182382 4 105 theme calculated 805:814 arg1 weight 826:831 calculated molecular weight 805:831 calculated molecular weight of glycans and their fragment ions 805:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 0 106 theme intact 32:37 arg1 N-glycopeptides 39:53 intact N-glycopeptides 32:53 intact N-glycopeptides 32:53 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 12 107 theme matches 2091:2097 arg1 probability 2069:2079 a probability 2067:2079 a probability of random matches less than 1% 2067:2110 The reliability of this strategy was carefully evaluated which showed a probability of random matches less than 1%. 25182382 5 108 theme same 1031:1034 arg1 enrichment 1042:1051 the same HILIC enrichment 1027:1051 the same HILIC enrichment 1027:1051 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 2 109 theme complex 474:480 arg1 samples 491:497 complex proteome samples 474:497 complex proteome samples 474:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 7 110 theme intact 1392:1397 arg1 N-glycopeptides 1399:1413 2249 unique intact N-glycopeptides 1380:1413 2249 unique intact N-glycopeptides 1380:1413 This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. 25182382 0 111 theme automated 64:72 arg1 method 89:94 an automated glycoproteomic method 61:94 an automated glycoproteomic method 61:94 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 6 112 theme N-glycopeptides 1223:1237 arg1 identification 1194:1207 the identification 1190:1207 the identification of the intact N-glycopeptides 1190:1237 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 6 113 gly N-glycopeptides 1223:1237 arg2 N-glycopeptides 1223:1237 the intact N-glycopeptides 1212:1237 the intact N-glycopeptides 1212:1237 The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. 25182382 2 114 theme computational 371:383 arg1 strategy 385:392 an analytical and computational strategy 353:392 an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 353:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 0 115 gly N-glycopeptides 39:53 arg2 N-glycopeptides 39:53 intact N-glycopeptides 32:53 intact N-glycopeptides 32:53 Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method. 25182382 11 116 theme strong 1918:1923 arg1 correlation 1925:1935 The strong correlation 1914:1935 The strong correlation of retention times 1914:1954 The strong correlation of retention times effectively filtered out random matches. 25182382 4 117 theme ions 863:866 arg1 weight 826:831 calculated molecular weight 805:831 calculated molecular weight of glycans and their fragment ions 805:866 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 2 118 theme analytical 356:365 arg1 strategy 385:392 an analytical and computational strategy 353:392 an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 353:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 8 119 theme site-specific 1457:1469 arg1 N-glycans 1471:1479 1769 site-specific N-glycans 1452:1479 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations 1452:1566 These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations. 25182382 5 120 theme LC-MS/MS 1083:1090 arg1 analysis 1092:1099 a separated LC-MS/MS analysis 1071:1099 a separated LC-MS/MS analysis 1071:1099 The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. 25182382 4 121 theme glycopeptides 694:706 arg1 spectra 676:682 the spectra 672:682 the spectra of intact glycopeptides 672:706 The Y1 ion (peptide+GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. 25182382 12 122 dep probability 2069:2079 arg1 % 2110:2110 % 2110:2110 % 2110:2110 The reliability of this strategy was carefully evaluated which showed a probability of random matches less than 1%. 25182382 2 123 theme high 402:405 arg1 characterization 418:433 the high throughput characterization 398:433 the high throughput characterization of intact N-glycopeptides derived from complex proteome samples 398:497 In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. 25182382 1 124 theme intact 284:289 arg1 glycopeptides 291:303 intact glycopeptides 284:303 intact glycopeptides 284:303 The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. 25188817 4 0 gly glycosylation 604:616 arg2 site 618:621 the glycosylation site 600:621 the glycosylation site 600:621 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 7 1 from sites 1093:1097 arg1 β-hairpins 1065:1074 compact β-hairpins 1057:1074 compact β-hairpins at glycosylation sites 1057:1097 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 4 2 theme stacking 645:652 arg1 interactions 654:665 stacking interactions 645:665 stacking interactions with the aromatic Trp residue 645:695 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 6 3 theme protein-glycan 919:932 arg1 interactions 934:945 the intraprotein and protein-glycan interactions 898:945 the intraprotein and protein-glycan interactions of natural glycoproteins 898:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 6 4 theme intraprotein 902:913 arg1 interactions 934:945 the intraprotein and protein-glycan interactions 898:945 the intraprotein and protein-glycan interactions of natural glycoproteins 898:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 8 5 dep glycosylation 1173:1185 arg1 code 1187:1190 code 1187:1190 code 1187:1190 In summary, these studies elucidate basic biophysical principles for the glycosylation code. 25188817 0 6 dep glycosylation 16:28 arg1 code 30:33 code 30:33 code 30:33 Deciphering the glycosylation code. 25188817 1 7 theme impact 79:84 arg1 folding 99:105 impact glycoprotein folding 79:105 impact glycoprotein folding 79:105 Asparagine-linked carbohydrates profoundly impact glycoprotein folding, stability, and structure. 25188817 3 8 from simulations 299:309 arg1 solvent 323:329 explicit solvent 314:329 explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 314:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 5 9 theme simulated 726:734 arg1 ensembles 736:744 our simulated ensembles 722:744 our simulated ensembles 722:744 In addition, we compare our simulated ensembles with the known structures for full length glycoproteins. 25188817 1 10 theme glycoprotein 86:97 arg1 folding 99:105 impact glycoprotein folding 79:105 impact glycoprotein folding 79:105 Asparagine-linked carbohydrates profoundly impact glycoprotein folding, stability, and structure. 25188817 3 11 theme glycosylation 377:389 arg1 impact 367:372 the impact 363:372 the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 363:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 3 12 theme replica 263:269 arg1 dynamics 290:297 detailed replica exchange molecular dynamics 254:297 detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 254:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 7 13 theme glycosylation 1079:1091 arg1 sites 1093:1097 glycosylation sites 1079:1097 glycosylation sites 1079:1097 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 2 14 theme glycosylation 148:160 arg1 " 166:166 the "glycosylation code" 143:166 the "glycosylation code" that relates these effects to protein sequence 143:213 However, the "glycosylation code" that relates these effects to protein sequence remains unsolved. 25188817 3 15 theme exchange 271:278 arg1 dynamics 290:297 detailed replica exchange molecular dynamics 254:297 detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 254:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 6 16 theme remarkable 867:876 arg1 consistency 878:888 a remarkable consistency 865:888 a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins 865:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 3 17 theme sequon 422:427 arg1 Pro-Asn-Gly/Ala-Thr-Trp/Ala 429:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 4 18 theme steric 544:549 arg1 crowding 551:558 steric crowding 544:558 steric crowding by the side chain immediately following the glycosylation site 544:621 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 5 19 theme full 776:779 arg1 glycoproteins 788:800 full length glycoproteins 776:800 full length glycoproteins 776:800 In addition, we compare our simulated ensembles with the known structures for full length glycoproteins. 25188817 3 20 theme detailed 254:261 arg1 dynamics 290:297 detailed replica exchange molecular dynamics 254:297 detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 254:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 7 21 theme conformations 1039:1051 arg1 significance 1011:1022 the significance 1007:1022 the significance of left-handed conformations for compact β-hairpins at glycosylation sites 1007:1097 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 5 22 gly glycoproteins 788:800 arg1 glycoproteins 788:800 full length glycoproteins 776:800 full length glycoproteins 776:800 In addition, we compare our simulated ensembles with the known structures for full length glycoproteins. 25188817 8 23 theme biophysical 1142:1152 arg1 principles 1154:1163 basic biophysical principles 1136:1163 basic biophysical principles for the glycosylation code 1136:1190 In summary, these studies elucidate basic biophysical principles for the glycosylation code. 25188817 3 24 theme simulations 299:309 arg1 dynamics 290:297 detailed replica exchange molecular dynamics 254:297 detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 254:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 8 25 theme basic 1136:1140 arg1 principles 1154:1163 basic biophysical principles 1136:1163 basic biophysical principles for the glycosylation code 1136:1190 In summary, these studies elucidate basic biophysical principles for the glycosylation code. 25188817 3 26 theme central 414:420 arg1 Pro-Asn-Gly/Ala-Thr-Trp/Ala 429:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 2 27 dep " 166:166 arg1 code 162:165 code 162:165 code 162:165 However, the "glycosylation code" that relates these effects to protein sequence remains unsolved. 25188817 3 28 theme molecular 280:288 arg1 dynamics 290:297 detailed replica exchange molecular dynamics 254:297 detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 254:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 4 29 theme glycosylation 504:516 arg1 effects 493:499 the effects 489:499 the effects of glycosylation 489:516 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 4 29 theme glycosylation 504:516 arg1 sensitive 531:539 sensitive 531:539 sensitive 531:539 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 6 30 gly glycoproteins 958:970 arg1 glycoproteins 958:970 natural glycoproteins 950:970 natural glycoproteins 950:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 5 31 theme length 781:786 arg1 glycoproteins 788:800 full length glycoproteins 776:800 full length glycoproteins 776:800 In addition, we compare our simulated ensembles with the known structures for full length glycoproteins. 25188817 6 32 theme glycoproteins 958:970 arg1 interactions 934:945 the intraprotein and protein-glycan interactions 898:945 the intraprotein and protein-glycan interactions of natural glycoproteins 898:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 6 33 theme natural 950:956 arg1 glycoproteins 958:970 natural glycoproteins 950:970 natural glycoproteins 950:970 These structures corroborate the simulations and also suggest a remarkable consistency between the intraprotein and protein-glycan interactions of natural glycoproteins. 25188817 3 34 with peptides 396:403 arg1 Pro-Asn-Gly/Ala-Thr-Trp/Ala 429:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 410:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 4 35 theme side 567:570 arg1 chain 572:576 the side chain 563:576 the side chain immediately following the glycosylation site 563:621 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 4 36 theme glycosylation 604:616 arg1 site 618:621 the glycosylation site 600:621 the glycosylation site 600:621 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 4 37 with interactions 654:665 arg1 residue 689:695 the aromatic Trp residue 672:695 the aromatic Trp residue 672:695 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 3 38 theme explicit 314:321 arg1 solvent 323:329 explicit solvent 314:329 explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala 314:455 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 7 39 theme compact 1057:1063 arg1 β-hairpins 1065:1074 compact β-hairpins 1057:1074 compact β-hairpins at glycosylation sites 1057:1097 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 7 40 theme left-handed 1027:1037 arg1 conformations 1039:1051 left-handed conformations 1027:1051 left-handed conformations 1027:1051 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 4 41 theme aromatic 676:683 arg1 residue 689:695 the aromatic Trp residue 672:695 the aromatic Trp residue 672:695 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25188817 5 42 theme known 755:759 arg1 structures 761:770 the known structures 751:770 the known structures for full length glycoproteins 751:800 In addition, we compare our simulated ensembles with the known structures for full length glycoproteins. 25188817 2 43 theme protein 198:204 arg1 sequence 206:213 protein sequence 198:213 protein sequence 198:213 However, the "glycosylation code" that relates these effects to protein sequence remains unsolved. 25188817 1 44 theme Asparagine-linked 36:52 arg1 carbohydrates 54:66 Asparagine-linked carbohydrates 36:66 Asparagine-linked carbohydrates 36:66 Asparagine-linked carbohydrates profoundly impact glycoprotein folding, stability, and structure. 25188817 3 45 from solvent 323:329 arg1 simulations 299:309 simulations 299:309 simulations 299:309 We report atomically detailed replica exchange molecular dynamics simulations in explicit solvent that systematically investigate the impact of glycosylation upon peptides with the central sequon Pro-Asn-Gly/Ala-Thr-Trp/Ala. 25188817 1 46 gly glycoprotein 86:97 arg1 glycoprotein 86:97 impact glycoprotein folding 79:105 impact glycoprotein folding 79:105 Asparagine-linked carbohydrates profoundly impact glycoprotein folding, stability, and structure. 25188817 7 47 gly glycosylation 1079:1091 arg2 sites 1093:1097 glycosylation sites 1079:1097 glycosylation sites 1079:1097 Moreover, our analysis highlights the significance of left-handed conformations for compact β-hairpins at glycosylation sites. 25188817 1 48 link Asparagine-linked 36:52 arg1 carbohydrates 54:66 Asparagine-linked carbohydrates 36:66 Asparagine-linked carbohydrates 36:66 Asparagine-linked carbohydrates profoundly impact glycoprotein folding, stability, and structure. 25188817 4 49 theme Trp 685:687 arg1 residue 689:695 the aromatic Trp residue 672:695 the aromatic Trp residue 672:695 These simulations suggest that the effects of glycosylation may be quite sensitive to steric crowding by the side chain immediately following the glycosylation site but less sensitive to stacking interactions with the aromatic Trp residue. 25029481 2 0 from profiling 196:204 arg1 sections 229:236 tissue sections 222:236 tissue sections 222:236 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 0 1 link N-linked 11:18 arg1 glycans 20:26 N-linked glycans 11:26 N-linked glycans from formalin-fixed paraffin-embedded tissue sections 11:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 4 2 from analysis 662:669 arg1 tissues 729:735 formalin-fixed paraffin-embedded (FFPE) tissues 689:735 formalin-fixed paraffin-embedded (FFPE) tissues 689:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 8 3 theme mass 1269:1272 arg1 spectrometry 1274:1285 mass spectrometry 1269:1285 The mass spectrometry images 1265:1292 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 2 4 theme molecular 332:340 arg1 light 381:385 light 381:385 light 381:385 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 4 theme molecular 332:340 arg1 N-glycans 291:299 tissue-specific and/or disease-associated N-glycans 249:299 tissue-specific and/or disease-associated N-glycans 249:299 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 4 theme molecular 332:340 arg1 signatures 342:351 molecular signatures 332:351 molecular signatures for diagnosis 332:365 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 5 5 theme N-linked 880:887 arg1 glycans 889:895 N-linked glycans 880:895 N-linked glycans from proteins by printing peptide-N-glycosidase F 880:945 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 6 dep sectioning 765:774 arg1 tissues 781:787 FFPE tissues 776:787 FFPE tissues 776:787 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 4 7 theme MSI 637:639 arg1 technique 641:649 an MSI technique 634:649 an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues 634:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 10 8 theme differential 1666:1677 arg1 expression 1679:1688 differential expression 1666:1688 differential expression in tumor versus normal brain tissues 1666:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 3 9 theme tissue 601:606 arg1 sections 608:615 tissue sections 601:615 tissue sections 601:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 5 10 theme tissue 853:858 arg1 proteins 860:867 tissue proteins 853:867 tissue proteins 853:867 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 0 11 from sections 73:80 arg1 Imaging 0:6 Imaging 0:6 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections 0:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 0 11 from sections 73:80 arg1 glycans 20:26 N-linked glycans 11:26 N-linked glycans from formalin-fixed paraffin-embedded tissue sections 11:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 5 12 theme mass 1075:1078 arg1 MALDI-MS 1094:1101 MALDI-MS 1094:1101 MALDI-MS 1094:1101 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 12 theme mass 1075:1078 arg1 spectrometry 1080:1091 matrix-assisted laser desorption/ionization mass spectrometry 1031:1091 matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) 1031:1102 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 13 theme peptide-N-glycosidase 923:943 arg1 F 945:945 printing peptide-N-glycosidase F 914:945 printing peptide-N-glycosidase F 914:945 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 8 14 from abundance 1326:1334 arg1 isocortex 1438:1446 isocortex 1438:1446 isocortex 1438:1446 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 14 from abundance 1326:1334 arg1 formation 1461:1469 hippocampal formation 1449:1469 hippocampal formation 1449:1469 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 14 from abundance 1326:1334 arg1 areas 1422:1426 different brain areas 1406:1426 different brain areas including isocortex, hippocampal formation, and brainstem 1406:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 14 from abundance 1326:1334 arg1 brainstem 1476:1484 brainstem 1476:1484 brainstem 1476:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 15 theme relative 1317:1324 arg1 abundance 1326:1334 the relative abundance 1313:1334 the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem 1313:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 9 16 theme NOD/SCID 1617:1624 arg1 mouse 1626:1630 a NOD/SCID mouse 1615:1630 a NOD/SCID mouse 1615:1630 Furthermore, glioblastoma tumor xenografts in a NOD/SCID mouse were imaged. 25029481 4 17 theme N-glycans 674:682 arg1 analysis 662:669 direct analysis 655:669 direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues 655:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 11 18 theme N-glycans 1784:1792 arg1 imaging 1773:1779 imaging 1773:1779 imaging of N-glycans 1773:1792 The MSI technique allows for imaging of N-glycans directly from FFPE sections. 25029481 7 19 theme brain 1250:1254 arg1 section 1256:1262 the mouse brain section 1240:1262 the mouse brain section 1240:1262 Forty-two N-glycans were analyzed from the mouse brain section. 25029481 4 20 theme formalin-fixed 689:702 arg1 tissues 729:735 formalin-fixed paraffin-embedded (FFPE) tissues 689:735 formalin-fixed paraffin-embedded (FFPE) tissues 689:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 0 21 theme MALDI 88:92 arg1 spectrometry 99:110 MALDI mass spectrometry 88:110 MALDI mass spectrometry 88:110 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 5 22 from proteins 902:909 arg1 glycans 889:895 N-linked glycans 880:895 N-linked glycans from proteins by printing peptide-N-glycosidase F 880:945 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 0 23 from Imaging 0:6 arg1 sections 73:80 formalin-fixed paraffin-embedded tissue sections 33:80 formalin-fixed paraffin-embedded tissue sections 33:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 9 24 theme glioblastoma 1582:1593 arg1 xenografts 1601:1610 glioblastoma tumor xenografts 1582:1610 glioblastoma tumor xenografts in a NOD/SCID mouse 1582:1630 Furthermore, glioblastoma tumor xenografts in a NOD/SCID mouse were imaged. 25029481 8 25 theme fucosylated 1373:1383 arg1 N-glycans 1393:1401 fucosylated complex N-glycans 1373:1401 fucosylated complex N-glycans 1373:1401 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 10 26 theme normal 1706:1711 arg1 tissues 1719:1725 tumor versus normal brain tissues 1693:1725 tissues 1719:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 5 27 link N-linked 880:887 arg1 glycans 889:895 N-linked glycans 880:895 N-linked glycans from proteins by printing peptide-N-glycosidase F 880:945 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 6 28 theme Brain 1105:1109 arg1 sections 1111:1118 Brain sections 1105:1118 Brain sections from a C57BL/6 mouse 1105:1139 Brain sections from a C57BL/6 mouse were imaged using this technique at a resolution of 100 μm. 25029481 8 29 theme N-glycans 1393:1401 arg1 abundance 1326:1334 the relative abundance 1313:1334 the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem 1313:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 10 30 theme tumor 1693:1697 arg1 tissues 1719:1725 tumor versus normal brain tissues 1693:1725 tissues 1719:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 8 31 theme different 1406:1414 arg1 isocortex 1438:1446 isocortex 1438:1446 isocortex 1438:1446 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 31 theme different 1406:1414 arg1 formation 1461:1469 hippocampal formation 1449:1469 hippocampal formation 1449:1469 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 31 theme different 1406:1414 arg1 areas 1422:1426 different brain areas 1406:1426 different brain areas including isocortex, hippocampal formation, and brainstem 1406:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 31 theme different 1406:1414 arg1 brainstem 1476:1484 brainstem 1476:1484 brainstem 1476:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 3 32 from histopathology 583:596 arg1 sections 608:615 tissue sections 601:615 tissue sections 601:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 3 33 theme powerful 471:478 arg1 imaging 452:458 Mass spectrometry imaging 434:458 Mass spectrometry imaging (MSI) 434:464 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 3 33 theme powerful 471:478 arg1 tool 480:483 a powerful tool 469:483 a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections 469:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 0 34 theme N-linked 11:18 arg1 glycans 20:26 N-linked glycans 11:26 N-linked glycans from formalin-fixed paraffin-embedded tissue sections 11:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 4 35 dep paraffin-embedded 704:720 arg1 FFPE 723:726 FFPE 723:726 FFPE 723:726 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 3 36 theme Mass 434:437 arg1 spectrometry 439:450 Mass spectrometry 434:450 Mass spectrometry imaging (MSI) 434:464 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 12 37 theme certain 1964:1970 arg1 structures 1985:1994 certain histological structures 1964:1994 certain histological structures 1964:1994 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 5 38 theme matrix-assisted 1031:1045 arg1 MALDI-MS 1094:1101 MALDI-MS 1094:1101 MALDI-MS 1094:1101 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 38 theme matrix-assisted 1031:1045 arg1 spectrometry 1080:1091 matrix-assisted laser desorption/ionization mass spectrometry 1031:1091 matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) 1031:1102 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 12 39 theme disease-associated 1883:1900 arg1 glycans 1902:1908 tissue-specific and/or disease-associated glycans 1860:1908 tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures 1860:1994 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 2 40 theme Direct 177:182 arg1 imaging 184:190 Direct imaging 177:190 Direct imaging 177:190 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 3 41 used used 499:502 arg2 tool 480:483 a powerful tool 469:483 a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections 469:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 3 41 used used 499:502 arg2 imaging 452:458 Mass spectrometry imaging 434:458 Mass spectrometry imaging (MSI) 434:464 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 0 42 theme paraffin-embedded 48:64 arg1 sections 73:80 formalin-fixed paraffin-embedded tissue sections 33:80 formalin-fixed paraffin-embedded tissue sections 33:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 5 43 theme desorption/ionization 1053:1073 arg1 MALDI-MS 1094:1101 MALDI-MS 1094:1101 MALDI-MS 1094:1101 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 43 theme desorption/ionization 1053:1073 arg1 spectrometry 1080:1091 matrix-assisted laser desorption/ionization mass spectrometry 1031:1091 matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) 1031:1102 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 8 44 theme hippocampal 1449:1459 arg1 formation 1461:1469 hippocampal formation 1449:1469 hippocampal formation 1449:1469 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 12 45 theme molecular 1933:1941 arg1 signatures 1943:1952 other molecular signatures 1927:1952 other molecular signatures 1927:1952 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 10 46 theme brain 1713:1717 arg1 tissues 1719:1725 tumor versus normal brain tissues 1693:1725 tissues 1719:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 12 47 theme tissue-specific 1860:1874 arg1 glycans 1902:1908 tissue-specific and/or disease-associated glycans 1860:1908 tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures 1860:1994 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 6 48 theme C57BL/6 1127:1133 arg1 mouse 1135:1139 a C57BL/6 mouse 1125:1139 a C57BL/6 mouse 1125:1139 Brain sections from a C57BL/6 mouse were imaged using this technique at a resolution of 100 μm. 25029481 6 49 theme μm 1197:1198 arg1 resolution 1179:1188 a resolution 1177:1188 a resolution of 100 μm 1177:1198 Brain sections from a C57BL/6 mouse were imaged using this technique at a resolution of 100 μm. 25029481 10 50 from expression 1679:1688 arg1 tissues 1719:1725 tumor versus normal brain tissues 1693:1725 tissues 1719:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 10 51 theme Several 1645:1651 arg1 glycans 1653:1659 Several glycans 1645:1659 Several glycans with differential expression in tumor versus normal brain tissues 1645:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 5 52 theme FFPE 776:779 arg1 tissues 781:787 FFPE tissues 776:787 FFPE tissues 776:787 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 4 53 theme direct 655:660 arg1 analysis 662:669 direct analysis 655:669 direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues 655:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 5 54 theme sections 832:839 arg1 deparaffinization 790:806 deparaffinization 790:806 deparaffinization 790:806 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 54 theme sections 832:839 arg1 rehydration 813:823 rehydration 813:823 rehydration of the sections 813:839 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 54 theme sections 832:839 arg1 sectioning 765:774 sectioning FFPE tissues, deparaffinization, and rehydration of the sections 765:839 sectioning 765:774 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 8 55 used used 1299:1302 arg2 images 1287:1292 The mass spectrometry images 1265:1292 The mass spectrometry images 1265:1292 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 56 theme spectrometry 1274:1285 arg1 images 1287:1292 The mass spectrometry images 1265:1292 The mass spectrometry images 1265:1292 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 2 57 theme tissue-specific 249:263 arg1 light 381:385 light 381:385 light 381:385 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 57 theme tissue-specific 249:263 arg1 N-glycans 291:299 tissue-specific and/or disease-associated N-glycans 249:299 tissue-specific and/or disease-associated N-glycans 249:299 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 57 theme tissue-specific 249:263 arg1 signatures 342:351 molecular signatures 332:351 molecular signatures for diagnosis 332:365 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 8 58 theme specific 1490:1497 arg1 glycans 1499:1505 specific glycans 1490:1505 specific glycans associated with different areas of the brain 1490:1550 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 3 59 theme underlying 572:581 arg1 histopathology 583:596 their underlying histopathology 566:596 their underlying histopathology in tissue sections 566:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 5 60 theme printing 914:921 arg1 F 945:945 printing peptide-N-glycosidase F 914:945 printing peptide-N-glycosidase F 914:945 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 8 61 theme different 1523:1531 arg1 areas 1533:1537 different areas 1523:1537 different areas of the brain 1523:1550 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 11 62 theme FFPE 1808:1811 arg1 sections 1813:1820 FFPE sections 1808:1820 FFPE sections 1808:1820 The MSI technique allows for imaging of N-glycans directly from FFPE sections. 25029481 7 63 theme mouse 1244:1248 arg1 section 1256:1262 the mouse brain section 1240:1262 the mouse brain section 1240:1262 Forty-two N-glycans were analyzed from the mouse brain section. 25029481 2 64 theme disease-associated 272:289 arg1 light 381:385 light 381:385 light 381:385 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 64 theme disease-associated 272:289 arg1 N-glycans 291:299 tissue-specific and/or disease-associated N-glycans 249:299 tissue-specific and/or disease-associated N-glycans 249:299 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 64 theme disease-associated 272:289 arg1 signatures 342:351 molecular signatures 332:351 molecular signatures for diagnosis 332:365 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 0 65 theme mass 94:97 arg1 spectrometry 99:110 MALDI mass spectrometry 88:110 MALDI mass spectrometry 88:110 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 9 66 from xenografts 1601:1610 arg1 mouse 1626:1630 a NOD/SCID mouse 1615:1630 a NOD/SCID mouse 1615:1630 Furthermore, glioblastoma tumor xenografts in a NOD/SCID mouse were imaged. 25029481 8 67 theme brain 1546:1550 arg1 areas 1533:1537 different areas 1523:1537 different areas of the brain 1523:1550 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 68 theme oligomannose 1339:1350 arg1 abundance 1326:1334 the relative abundance 1313:1334 the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem 1313:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 4 69 dep formalin-fixed 689:702 arg1 paraffin-embedded 704:720 paraffin-embedded 704:720 paraffin-embedded 704:720 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 8 70 theme nonfucosylated 1353:1366 arg1 abundance 1326:1334 the relative abundance 1313:1334 the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem 1313:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 7 71 theme Forty-two 1201:1209 arg1 N-glycans 1211:1219 Forty-two N-glycans 1201:1219 Forty-two N-glycans 1201:1219 Forty-two N-glycans were analyzed from the mouse brain section. 25029481 9 72 theme tumor 1595:1599 arg1 xenografts 1601:1610 glioblastoma tumor xenografts 1582:1610 glioblastoma tumor xenografts in a NOD/SCID mouse 1582:1630 Furthermore, glioblastoma tumor xenografts in a NOD/SCID mouse were imaged. 25029481 8 73 theme complex 1385:1391 arg1 N-glycans 1393:1401 fucosylated complex N-glycans 1373:1401 fucosylated complex N-glycans 1373:1401 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 2 74 theme functional 394:403 arg1 roles 405:409 the functional roles 390:409 the functional roles of these biomolecules 390:431 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 0 75 theme glycans 20:26 arg1 Imaging 0:6 Imaging 0:6 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections 0:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 4 76 from tissues 729:735 arg1 N-glycans 674:682 N-glycans 674:682 N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues 674:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 4 76 from tissues 729:735 arg1 analysis 662:669 direct analysis 655:669 direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues 655:735 Here, we report an MSI technique for direct analysis of N-glycans from formalin-fixed paraffin-embedded (FFPE) tissues. 25029481 1 77 theme Aberrant 113:120 arg1 glycosylation 122:134 Aberrant glycosylation 113:134 Aberrant glycosylation 113:134 Aberrant glycosylation is associated with most of the diseases. 25029481 11 78 theme MSI 1748:1750 arg1 technique 1752:1760 The MSI technique 1744:1760 The MSI technique 1744:1760 The MSI technique allows for imaging of N-glycans directly from FFPE sections. 25029481 0 79 theme formalin-fixed 33:46 arg1 sections 73:80 formalin-fixed paraffin-embedded tissue sections 33:80 formalin-fixed paraffin-embedded tissue sections 33:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 8 80 gly fucosylated 1373:1383 arg1 N-glycans 1393:1401 fucosylated complex N-glycans 1373:1401 fucosylated complex N-glycans 1373:1401 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 12 81 theme histological 1972:1983 arg1 structures 1985:1994 certain histological structures 1964:1994 certain histological structures 1964:1994 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 3 82 theme spectrometry 439:450 arg1 MSI 461:463 MSI 461:463 MSI 461:463 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 3 82 theme spectrometry 439:450 arg1 imaging 452:458 Mass spectrometry imaging 434:458 Mass spectrometry imaging (MSI) 434:464 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 3 82 theme spectrometry 439:450 arg1 tool 480:483 a powerful tool 469:483 a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections 469:615 Mass spectrometry imaging (MSI) is a powerful tool that has been used to correlate peptides, proteins, lipids, and metabolites with their underlying histopathology in tissue sections. 25029481 8 83 theme brain 1416:1420 arg1 isocortex 1438:1446 isocortex 1438:1446 isocortex 1438:1446 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 83 theme brain 1416:1420 arg1 formation 1461:1469 hippocampal formation 1449:1469 hippocampal formation 1449:1469 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 83 theme brain 1416:1420 arg1 areas 1422:1426 different brain areas 1406:1426 different brain areas including isocortex, hippocampal formation, and brainstem 1406:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 8 83 theme brain 1416:1420 arg1 brainstem 1476:1484 brainstem 1476:1484 brainstem 1476:1484 The mass spectrometry images were used to study the relative abundance of oligomannose, nonfucosylated, and fucosylated complex N-glycans in different brain areas including isocortex, hippocampal formation, and brainstem and specific glycans associated with different areas of the brain were identified. 25029481 2 84 theme biomolecules 420:431 arg1 roles 405:409 the functional roles 390:409 the functional roles of these biomolecules 390:431 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 0 85 theme tissue 66:71 arg1 sections 73:80 formalin-fixed paraffin-embedded tissue sections 33:80 formalin-fixed paraffin-embedded tissue sections 33:80 Imaging of N-linked glycans from formalin-fixed paraffin-embedded tissue sections using MALDI mass spectrometry. 25029481 12 86 theme other 1927:1931 arg1 signatures 1943:1952 other molecular signatures 1927:1952 other molecular signatures 1927:1952 This method can potentially identify tissue-specific and/or disease-associated glycans coexpressed with other molecular signatures or within certain histological structures. 25029481 5 87 theme laser 1047:1051 arg1 MALDI-MS 1094:1101 MALDI-MS 1094:1101 MALDI-MS 1094:1101 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 5 87 theme laser 1047:1051 arg1 spectrometry 1080:1091 matrix-assisted laser desorption/ionization mass spectrometry 1031:1091 matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) 1031:1102 This technique consists of sectioning FFPE tissues, deparaffinization, and rehydration of the sections, denaturing tissue proteins, releasing N-linked glycans from proteins by printing peptide-N-glycosidase F over the sections, spray-coating the tissue with matrix, and analyzing N-glycans by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). 25029481 2 88 theme tissue 222:227 arg1 sections 229:236 tissue sections 222:236 tissue sections 222:236 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 89 theme N-glycans 209:217 arg1 profiling 196:204 profiling 196:204 profiling 196:204 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 89 theme N-glycans 209:217 arg1 imaging 184:190 Direct imaging 177:190 Direct imaging 177:190 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 2 90 from imaging 184:190 arg1 sections 229:236 tissue sections 222:236 tissue sections 222:236 Direct imaging and profiling of N-glycans on tissue sections can reveal tissue-specific and/or disease-associated N-glycans, which not only could serve as molecular signatures for diagnosis but also shed light on the functional roles of these biomolecules. 25029481 10 91 with glycans 1653:1659 arg1 expression 1679:1688 differential expression 1666:1688 differential expression in tumor versus normal brain tissues 1666:1725 Several glycans with differential expression in tumor versus normal brain tissues were identified. 25029481 6 92 from mouse 1135:1139 arg1 sections 1111:1118 Brain sections 1105:1118 Brain sections from a C57BL/6 mouse 1105:1139 Brain sections from a C57BL/6 mouse were imaged using this technique at a resolution of 100 μm. 28493134 7 0 theme glycoprotein 1244:1255 arg1 purification 1257:1268 glycoprotein purification 1244:1268 glycoprotein purification after separation from producing cell lines 1244:1311 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 2 1 theme cell 354:357 arg1 lines 359:363 Several cell lines 346:363 Several cell lines 346:363 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 1 2 theme clinical 245:252 arg1 trials 254:259 clinical trials 245:259 clinical trials 245:259 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 3 from pharmaceuticals 315:329 arg1 market 338:343 the market 334:343 the market 334:343 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 2 4 theme Several 346:352 arg1 lines 359:363 Several cell lines 346:363 Several cell lines 346:363 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 6 5 theme glycosylation 1118:1130 arg1 analysis 1132:1139 glycosylation analysis 1118:1139 glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1118:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 7 6 gly glycoprotein 1244:1255 arg1 glycoprotein 1244:1255 glycoprotein purification 1244:1268 glycoprotein purification after separation from producing cell lines 1244:1311 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 6 7 theme Hamster 1195:1201 arg1 CHO 1182:1184 CHO 1182:1184 CHO (Chinese Hamster Ovary) cells 1182:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 6 7 theme Hamster 1195:1201 arg1 Ovary 1203:1207 Chinese Hamster Ovary 1187:1207 Chinese Hamster Ovary 1187:1207 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 1 8 from class 306:310 arg1 market 338:343 the market 334:343 the market 334:343 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 7 9 theme producing 1292:1300 arg1 lines 1307:1311 producing cell lines 1292:1311 producing cell lines 1292:1311 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 3 10 dep common 589:594 arg1 the 580:582 the 580:582 the 580:582 Glycosylation is the most common and complex (for both bioprocess engineering and quality control) of these modifications. 28493134 4 11 theme therapeutic 745:755 arg1 proteins 757:764 therapeutic proteins 745:764 therapeutic proteins 745:764 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 7 12 theme cell 1302:1305 arg1 lines 1307:1311 producing cell lines 1292:1311 producing cell lines 1292:1311 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 7 13 theme structural 1355:1364 arg1 characterization 1366:1381 fine structural characterization 1350:1381 fine structural characterization 1350:1381 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 7 14 theme mass 1391:1394 arg1 techniques 1409:1418 mass spectrometry techniques 1391:1418 mass spectrometry techniques 1391:1418 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 2 15 theme post-translational 522:539 arg1 modifications 541:553 human-like post-translational modifications 511:553 human-like post-translational modifications (PTMs) 511:560 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 2 15 theme post-translational 522:539 arg1 PTMs 556:559 PTMs 556:559 PTMs 556:559 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 2 16 theme biopharmaceutical 383:399 arg1 processes 401:409 biopharmaceutical processes 383:409 biopharmaceutical processes 383:409 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 4 17 dep way 883:885 arg1 interacts 892:900 interacts 892:900 interacts with patients' cells 892:921 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 1 18 theme last 82:85 arg1 decades 87:93 the last decades 78:93 the last decades 78:93 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 7 19 theme N-glycan 1314:1321 arg1 release 1323:1329 N-glycan release 1314:1329 N-glycan release 1314:1329 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 0 20 theme Glycosylation 0:12 arg1 Analysis 14:21 Glycosylation Analysis 0:21 Glycosylation Analysis of Therapeutic Glycoproteins 0:50 Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells. 28493134 6 21 gly glycoproteins 1156:1168 arg1 glycoproteins 1156:1168 therapeutic glycoproteins 1144:1168 therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1144:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 6 22 theme Chinese 1187:1193 arg1 CHO 1182:1184 CHO 1182:1184 CHO (Chinese Hamster Ovary) cells 1182:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 6 22 theme Chinese 1187:1193 arg1 Ovary 1203:1207 Chinese Hamster Ovary 1187:1207 Chinese Hamster Ovary 1187:1207 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 1 23 theme large 180:184 arg1 number 186:191 a large number 178:191 a large number of new therapeutic entities 178:219 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 0 24 theme Therapeutic 26:36 arg1 Glycoproteins 38:50 Therapeutic Glycoproteins 26:50 Therapeutic Glycoproteins 26:50 Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells. 28493134 7 25 from lines 1307:1311 arg1 purification 1257:1268 glycoprotein purification 1244:1268 glycoprotein purification after separation from producing cell lines 1244:1311 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 7 25 from lines 1307:1311 arg1 release 1323:1329 N-glycan release 1314:1329 N-glycan release 1314:1329 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 7 25 from lines 1307:1311 arg1 variants 1341:1348 their variants 1335:1348 their variants 1335:1348 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 5 26 theme other 962:966 arg1 PTMs 968:971 other PTMs 962:971 other PTMs 962:971 As a consequence, glycosylation (like other PTMs) needs to be regulated and accurately analyzed during biopharmaceutical production. 28493134 7 27 theme fine 1350:1353 arg1 characterization 1366:1381 fine structural characterization 1350:1381 fine structural characterization 1350:1381 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 6 28 theme therapeutic 1144:1154 arg1 glycoproteins 1156:1168 therapeutic glycoproteins 1144:1168 therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1144:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 7 29 dep purification 1257:1268 arg1 characterization 1366:1381 fine structural characterization 1350:1381 fine structural characterization 1350:1381 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 6 30 theme CHO 1182:1184 arg1 cells 1210:1214 CHO (Chinese Hamster Ovary) cells 1182:1214 CHO (Chinese Hamster Ovary) cells 1182:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 6 31 gly glycosylation 1118:1130 arg1 glycoproteins 1156:1168 therapeutic glycoproteins 1144:1168 therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1144:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 4 32 theme glycosylation 708:720 arg1 pattern 722:728 the desired glycosylation pattern 696:728 the desired glycosylation pattern 696:728 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 1 33 from market 338:343 arg1 class 306:310 the most promising class 287:310 the most promising class of pharmaceuticals on the market 287:343 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 33 from market 338:343 arg1 biologics 274:282 biologics 274:282 biologics 274:282 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 34 theme new 196:198 arg1 entities 212:219 new therapeutic entities 196:219 new therapeutic entities 196:219 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 2 35 theme mammalian 415:423 arg1 cells 425:429 mammalian cells 415:429 mammalian cells 415:429 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 0 36 theme Glycoproteins 38:50 arg1 Analysis 14:21 Glycosylation Analysis 0:21 Glycosylation Analysis of Therapeutic Glycoproteins 0:50 Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells. 28493134 1 37 theme promising 296:304 arg1 class 306:310 the most promising class 287:310 the most promising class of pharmaceuticals on the market 287:343 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 37 theme promising 296:304 arg1 biologics 274:282 biologics 274:282 biologics 274:282 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 5 38 theme biopharmaceutical 1027:1043 arg1 production 1045:1054 biopharmaceutical production 1027:1054 biopharmaceutical production 1027:1054 As a consequence, glycosylation (like other PTMs) needs to be regulated and accurately analyzed during biopharmaceutical production. 28493134 1 39 theme therapeutic 200:210 arg1 entities 212:219 new therapeutic entities 196:219 new therapeutic entities 196:219 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 0 40 theme CHO 64:66 arg1 Cells 68:72 CHO Cells 64:72 CHO Cells 64:72 Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells. 28493134 4 41 dep impact 776:781 arg1 stability 797:805 stability 797:805 stability 797:805 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 4 41 dep impact 776:781 arg1 driving 840:846 driving 840:846 driving molecular processes 840:866 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 4 41 dep impact 776:781 arg1 safety 822:827 safety 822:827 safety 822:827 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 4 41 dep impact 776:781 arg1 half-life 808:816 half-life 808:816 half-life 808:816 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 2 42 theme fewer 461:465 arg1 problems 467:474 fewer problems 461:474 fewer problems for immunogenicity 461:493 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 4 43 theme molecular 848:856 arg1 processes 858:866 molecular processes 848:866 molecular processes 848:866 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 7 44 theme spectrometry 1396:1407 arg1 techniques 1409:1418 mass spectrometry techniques 1391:1418 mass spectrometry techniques 1391:1418 This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques. 28493134 1 45 theme pharmaceuticals 315:329 arg1 class 306:310 the most promising class 287:310 the most promising class of pharmaceuticals on the market 287:343 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 45 theme pharmaceuticals 315:329 arg1 biologics 274:282 biologics 274:282 biologics 274:282 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 4 46 theme desired 700:706 arg1 pattern 722:728 the desired glycosylation pattern 696:728 the desired glycosylation pattern 696:728 Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. 28493134 1 47 theme approved 110:117 arg1 drugs 140:144 approved therapeutic proteins drugs 110:144 approved therapeutic proteins drugs 110:144 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 1 48 theme entities 212:219 arg1 number 186:191 a large number 178:191 a large number of new therapeutic entities 178:219 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 3 49 theme modifications 671:683 arg1 common 589:594 common 589:594 common 589:594 Glycosylation is the most common and complex (for both bioprocess engineering and quality control) of these modifications. 28493134 3 50 theme quality 645:651 arg1 control 653:659 quality control 645:659 quality control 645:659 Glycosylation is the most common and complex (for both bioprocess engineering and quality control) of these modifications. 28493134 1 51 theme therapeutic 119:129 arg1 drugs 140:144 approved therapeutic proteins drugs 110:144 approved therapeutic proteins drugs 110:144 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 6 52 theme glycoproteins 1156:1168 arg1 analysis 1132:1139 glycosylation analysis 1118:1139 glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1118:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 2 53 theme human-like 511:520 arg1 modifications 541:553 human-like post-translational modifications 511:553 human-like post-translational modifications (PTMs) 511:560 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 2 53 theme human-like 511:520 arg1 PTMs 556:559 PTMs 556:559 PTMs 556:559 Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). 28493134 3 54 theme bioprocess 618:627 arg1 engineering 629:639 bioprocess engineering 618:639 bioprocess engineering 618:639 Glycosylation is the most common and complex (for both bioprocess engineering and quality control) of these modifications. 28493134 1 55 theme proteins 131:138 arg1 drugs 140:144 approved therapeutic proteins drugs 110:144 approved therapeutic proteins drugs 110:144 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28493134 6 56 theme analytical 1092:1101 arg1 approaches 1103:1112 the analytical approaches 1088:1112 the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells 1088:1214 Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. 28493134 1 57 theme drugs 140:144 arg1 number 100:105 the number 96:105 the number of approved therapeutic proteins drugs 96:144 In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. 28803068 7 0 theme Cleaved 1188:1194 arg1 PARP 1196:1199 Cleaved PARP 1188:1199 Cleaved PARP 1188:1199 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 3 1 dep signalling 580:589 arg1 procaspase-3 626:637 procaspase-3 626:637 procaspase-3 626:637 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 1 dep signalling 580:589 arg1 FADD 640:643 FADD 640:643 FADD 640:643 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 1 dep signalling 580:589 arg1 caspase-3 664:672 caspase-3 664:672 caspase-3 664:672 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 1 dep signalling 580:589 arg1 H3 622:623 histone H3 614:623 histone H3 614:623 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 1 dep signalling 580:589 arg1 PARP 654:657 cleaved PARP 646:657 cleaved PARP 646:657 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 1 dep signalling 580:589 arg1 blot 600:603 Western blot 592:603 Western blot of Cdk2 592:611 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 10 2 theme NF-κB 1562:1566 arg1 activation 1548:1557 activation 1548:1557 activation of NF-κB 1548:1566 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 0 3 theme cancer 98:103 arg1 cells 105:109 ovarian cancer cells 90:109 ovarian cancer cells 90:109 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 6 4 with pattern 1113:1119 arg1 sialylation 1153:1163 some terminal sialylation 1139:1163 some terminal sialylation 1139:1163 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 6 4 with pattern 1113:1119 arg1 fucosylation 1174:1185 core fucosylation 1169:1185 core fucosylation 1169:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 4 5 from present 780:786 arg1 hIFNγ 808:812 HEK293-expressed hIFNγ 791:812 HEK293-expressed hIFNγ (hIFNγ-HEK) 791:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 4 5 from present 780:786 arg1 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 6 6 gly sialylation 1153:1163 arg1 pattern 1113:1119 the N-glycosylation pattern 1093:1119 the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation 1093:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 10 7 theme therapeutic 1592:1602 arg1 target 1604:1609 a novel therapeutic target 1584:1609 a novel therapeutic target 1584:1609 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 10 7 theme therapeutic 1592:1602 arg1 presence 1485:1492 The presence 1481:1492 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, 1481:1567 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 5 8 theme dose-independent 1022:1037 arg1 manner 1039:1044 a dose-independent manner 1020:1044 a dose-independent manner 1020:1044 PEO1 was more sensitive to hIFNγ than SKOV3, but responses were dose-dependent and expression platform/glycosylation status-independent, whereas SKOV3 responded to mammalian-expressed hIFNγ in a dose-independent manner, only. 28803068 3 9 dep hIFNγ-CHO 446:454 arg1 glycosylated/de-glycosylated 468:495 glycosylated/de-glycosylated 468:495 glycosylated/de-glycosylated 468:495 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 2 10 theme system 313:318 arg1 effect 274:279 the effect 270:279 the effect of glycosylation and expression system of recombinant hIFNγ 270:339 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 5 11 theme mammalian-expressed 991:1009 arg1 hIFNγ 1011:1015 mammalian-expressed hIFNγ 991:1015 mammalian-expressed hIFNγ 991:1015 PEO1 was more sensitive to hIFNγ than SKOV3, but responses were dose-dependent and expression platform/glycosylation status-independent, whereas SKOV3 responded to mammalian-expressed hIFNγ in a dose-independent manner, only. 28803068 2 12 theme expression 302:311 arg1 system 313:318 expression system 302:318 expression system 302:318 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 4 13 theme Interaction 701:711 arg1 Chromatography 720:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 7 14 theme cell 1251:1254 arg1 line 1256:1259 either cell line 1244:1259 line 1256:1259 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 8 15 dep line 1398:1401 arg1 either 1386:1391 either 1386:1391 either 1386:1391 In conclusion, hIFNγ did not induce apoptosis in either cell line. 28803068 4 16 theme present 780:786 arg1 glycans 772:778 N-linked glycans 763:778 N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK) 763:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 6 17 theme hIFNγ-HEK 1124:1132 arg1 pattern 1113:1119 the N-glycosylation pattern 1093:1119 the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation 1093:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 1 18 theme immune 178:183 arg1 responses 185:193 immune responses 178:193 immune responses 178:193 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 3 19 theme coli- 409:413 arg1 efficacy 394:401 The efficacy 390:401 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) 390:673 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 20 from efficacy 394:401 arg1 signalling 580:589 apoptotic signalling 570:589 apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) 570:673 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 20 from efficacy 394:401 arg1 cytostasis 501:510 cytostasis 501:510 cytostasis 501:510 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 20 from efficacy 394:401 arg1 death 518:522 cell death 513:522 cell death (MTT, and Guava-ViaCount® flow-cytometry) 513:564 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 2 21 theme glycosylation 284:296 arg1 effect 274:279 the effect 270:279 the effect of glycosylation and expression system of recombinant hIFNγ 270:339 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 6 22 theme terminal 1144:1151 arg1 sialylation 1153:1163 some terminal sialylation 1139:1163 some terminal sialylation 1139:1163 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 0 23 theme therapeutic 9:19 arg1 efficacy 21:28 therapeutic efficacy 9:28 therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells 9:109 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 4 24 attach present 780:786 arg1 hIFNγ 808:812 HEK293-expressed hIFNγ 791:812 HEK293-expressed hIFNγ (hIFNγ-HEK) 791:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 4 24 attach present 780:786 arg2 glycans 772:778 N-linked glycans 763:778 N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK) 763:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 4 24 attach present 780:786 arg1 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 6 25 gly fucosylation 1174:1185 arg1 pattern 1113:1119 the N-glycosylation pattern 1093:1119 the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation 1093:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 10 26 theme novel 1586:1590 arg1 target 1604:1609 a novel therapeutic target 1584:1609 a novel therapeutic target 1584:1609 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 10 26 theme novel 1586:1590 arg1 presence 1485:1492 The presence 1481:1492 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, 1481:1567 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 3 27 dep death 518:522 arg1 flow-cytometry 550:563 flow-cytometry 550:563 flow-cytometry 550:563 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 27 dep death 518:522 arg1 MTT 525:527 MTT 525:527 MTT 525:527 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 28 theme hIFNγ 439:443 arg1 efficacy 394:401 The efficacy 390:401 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) 390:673 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 6 29 theme Complex-type 1053:1064 arg1 oligosaccharides 1066:1081 Complex-type oligosaccharides 1053:1081 Complex-type oligosaccharides 1053:1081 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 7 30 located detected 1232:1239 arg2 PARP 1196:1199 Cleaved PARP 1188:1199 Cleaved PARP 1188:1199 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 7 30 located detected 1232:1239 arg1 line 1256:1259 either cell line 1244:1259 line 1256:1259 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 7 30 located detected 1232:1239 arg2 caspase-3 1213:1221 cleaved caspase-3 1205:1221 cleaved caspase-3 1205:1221 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 1 31 theme Human 112:116 arg1 hIFNγ 136:140 hIFNγ 136:140 hIFNγ 136:140 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 1 31 theme Human 112:116 arg1 gamma 129:133 Human interferon gamma 112:133 Human interferon gamma (hIFNγ) 112:141 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 10 32 theme FADD 1497:1500 arg1 target 1604:1609 a novel therapeutic target 1584:1609 a novel therapeutic target 1584:1609 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 10 32 theme FADD 1497:1500 arg1 presence 1485:1492 The presence 1481:1492 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, 1481:1567 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 10 33 from presence 1485:1492 arg1 SKOV3 1505:1509 SKOV3 1505:1509 SKOV3 1505:1509 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 6 34 gly N-glycosylation 1097:1111 arg1 hIFNγ-HEK 1124:1132 hIFNγ-HEK 1124:1132 hIFNγ-HEK 1124:1132 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 2 35 theme recombinant 323:333 arg1 hIFNγ 335:339 recombinant hIFNγ 323:339 recombinant hIFNγ 323:339 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 36 theme hIFNγ 335:339 arg1 effect 274:279 the effect 270:279 the effect of glycosylation and expression system of recombinant hIFNγ 270:339 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 3 37 theme apoptotic 570:578 arg1 signalling 580:589 apoptotic signalling 570:589 apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) 570:673 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 6 38 theme N-glycosylation 1097:1111 arg1 pattern 1113:1119 the N-glycosylation pattern 1093:1119 the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation 1093:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 0 39 theme expressed-recombinant 43:63 arg1 gamma 76:80 mammalian expressed-recombinant interferon gamma 33:80 mammalian expressed-recombinant interferon gamma 33:80 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 3 40 theme cell 513:516 arg1 death 518:522 cell death 513:522 cell death (MTT, and Guava-ViaCount® flow-cytometry) 513:564 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 41 dep hIFNγ 439:443 arg1 hIFNγ-CHO 446:454 hIFNγ-CHO 446:454 hIFNγ-CHO 446:454 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 41 dep hIFNγ 439:443 arg1 HEK293 460:465 HEK293 460:465 HEK293 460:465 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 10 42 attach presence 1485:1492 arg1 SKOV3 1505:1509 SKOV3 1505:1509 SKOV3 1505:1509 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 10 42 attach presence 1485:1492 arg2 FADD 1497:1500 FADD 1497:1500 FADD 1497:1500 The presence of FADD in SKOV3, which may inhibit apoptosis through activation of NF-κB, could serve as a novel therapeutic target. 28803068 0 43 theme mammalian 33:41 arg1 gamma 76:80 mammalian expressed-recombinant interferon gamma 33:80 mammalian expressed-recombinant interferon gamma 33:80 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 3 44 theme mammalian-expressed 419:437 arg1 hIFNγ 439:443 mammalian-expressed hIFNγ 419:443 mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) 419:496 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 45 gly glycosylated/de-glycosylated 468:495 arg1 hIFNγ-CHO 446:454 hIFNγ-CHO 446:454 hIFNγ-CHO 446:454 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 1 46 theme anti-cancer 218:228 arg1 biotherapeutic 230:243 an anti-cancer biotherapeutic 215:243 an anti-cancer biotherapeutic 215:243 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 1 46 theme anti-cancer 218:228 arg1 promise 204:210 promise 204:210 promise 204:210 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 0 47 theme gamma 76:80 arg1 efficacy 21:28 therapeutic efficacy 9:28 therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells 9:109 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 8 48 theme cell 1393:1396 arg1 line 1398:1401 either cell line 1386:1401 line 1398:1401 In conclusion, hIFNγ did not induce apoptosis in either cell line. 28803068 3 49 theme Western 592:598 arg1 blot 600:603 Western blot 592:603 Western blot of Cdk2 592:611 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 6 50 theme core 1169:1172 arg1 fucosylation 1174:1185 core fucosylation 1169:1185 core fucosylation 1169:1185 Complex-type oligosaccharides dominated the N-glycosylation pattern of hIFNγ-HEK with some terminal sialylation and core fucosylation. 28803068 4 51 theme Hydrophilic 689:699 arg1 Chromatography 720:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 0 52 theme interferon 65:74 arg1 gamma 76:80 mammalian expressed-recombinant interferon gamma 33:80 mammalian expressed-recombinant interferon gamma 33:80 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 9 53 theme hIFNγ 1425:1429 arg1 death 1446:1450 hIFNγ increased cell death 1425:1450 hIFNγ increased cell death 1425:1450 Mammalian- expressed hIFNγ increased cell death in the drug-resistant SKOV3. 28803068 7 54 dep line 1256:1259 arg1 either 1244:1249 either 1244:1249 either 1244:1249 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 2 55 theme cell 362:365 arg1 PEO1 374:377 PEO1 374:377 PEO1 374:377 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 55 theme cell 362:365 arg1 lines 367:371 ovarian carcinoma cell lines 344:371 ovarian carcinoma cell lines 344:371 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 55 theme cell 362:365 arg1 SKOV3 383:387 SKOV3 383:387 SKOV3 383:387 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 4 56 theme Liquid 713:718 arg1 Chromatography 720:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography 689:733 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 9 57 theme increased 1431:1439 arg1 death 1446:1450 hIFNγ increased cell death 1425:1450 hIFNγ increased cell death 1425:1450 Mammalian- expressed hIFNγ increased cell death in the drug-resistant SKOV3. 28803068 7 58 theme cleaved 1205:1211 arg1 caspase-3 1213:1221 cleaved caspase-3 1205:1221 cleaved caspase-3 1205:1221 Cleaved PARP and cleaved caspase-3 were not detected in either cell line, but FADD was expressed in SKOV3 with levels increased following treatment. 28803068 5 59 dep sensitive 841:849 arg1 SKOV3 865:869 SKOV3 865:869 SKOV3 865:869 PEO1 was more sensitive to hIFNγ than SKOV3, but responses were dose-dependent and expression platform/glycosylation status-independent, whereas SKOV3 responded to mammalian-expressed hIFNγ in a dose-independent manner, only. 28803068 4 60 theme HEK293-expressed 791:806 arg1 hIFNγ 808:812 HEK293-expressed hIFNγ 791:812 HEK293-expressed hIFNγ (hIFNγ-HEK) 791:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 4 60 theme HEK293-expressed 791:806 arg1 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 hIFNγ-HEK 815:823 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 3 61 theme Cdk2 608:611 arg1 procaspase-3 626:637 procaspase-3 626:637 procaspase-3 626:637 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 61 theme Cdk2 608:611 arg1 FADD 640:643 FADD 640:643 FADD 640:643 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 61 theme Cdk2 608:611 arg1 caspase-3 664:672 caspase-3 664:672 caspase-3 664:672 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 61 theme Cdk2 608:611 arg1 H3 622:623 histone H3 614:623 histone H3 614:623 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 61 theme Cdk2 608:611 arg1 PARP 654:657 cleaved PARP 646:657 cleaved PARP 646:657 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 61 theme Cdk2 608:611 arg1 blot 600:603 Western blot 592:603 Western blot of Cdk2 592:611 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 2 62 theme carcinoma 352:360 arg1 PEO1 374:377 PEO1 374:377 PEO1 374:377 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 62 theme carcinoma 352:360 arg1 lines 367:371 ovarian carcinoma cell lines 344:371 ovarian carcinoma cell lines 344:371 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 62 theme carcinoma 352:360 arg1 SKOV3 383:387 SKOV3 383:387 SKOV3 383:387 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 1 63 theme interferon 118:127 arg1 hIFNγ 136:140 hIFNγ 136:140 hIFNγ 136:140 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 1 63 theme interferon 118:127 arg1 gamma 129:133 Human interferon gamma 112:133 Human interferon gamma (hIFNγ) 112:141 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 9 64 theme cell 1441:1444 arg1 death 1446:1450 hIFNγ increased cell death 1425:1450 hIFNγ increased cell death 1425:1450 Mammalian- expressed hIFNγ increased cell death in the drug-resistant SKOV3. 28803068 4 65 theme glycans 772:778 arg1 structure 750:758 the structure 746:758 the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK) 746:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 3 66 theme cleaved 646:652 arg1 PARP 654:657 cleaved PARP 646:657 cleaved PARP 646:657 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 4 67 theme N-linked 763:770 arg1 glycans 772:778 N-linked glycans 763:778 N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK) 763:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 2 68 theme ovarian 344:350 arg1 PEO1 374:377 PEO1 374:377 PEO1 374:377 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 68 theme ovarian 344:350 arg1 lines 367:371 ovarian carcinoma cell lines 344:371 ovarian carcinoma cell lines 344:371 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 2 68 theme ovarian 344:350 arg1 SKOV3 383:387 SKOV3 383:387 SKOV3 383:387 This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. 28803068 4 69 link N-linked 763:770 arg1 glycans 772:778 N-linked glycans 763:778 N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK) 763:824 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 0 70 theme ovarian 90:96 arg1 cells 105:109 ovarian cancer cells 90:109 ovarian cancer cells 90:109 Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells. 28803068 4 71 from hIFNγ 808:812 arg1 present 780:786 present 780:786 present 780:786 Hydrophilic Interaction Liquid Chromatography determined the structure of N-linked glycans present in HEK293-expressed hIFNγ (hIFNγ-HEK). 28803068 1 72 theme tumour 151:156 arg1 cells 158:162 tumour cells 151:162 tumour cells 151:162 Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. 28803068 3 73 theme E. 406:407 arg1 coli- 409:413 E. coli- 406:413 E. coli- 406:413 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28803068 3 74 theme histone 614:620 arg1 H3 622:623 histone H3 614:623 histone H3 614:623 The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount® flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined. 28193043 6 0 theme N-linked 1196:1203 arg1 glycopeptides 1205:1217 labeled peptic N-linked glycopeptides 1181:1217 labeled peptic N-linked glycopeptides 1181:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 5 1 theme HDX 747:749 arg1 information 751:761 HDX information 747:761 HDX information 747:761 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 2 2 theme oligosaccharide 369:383 arg1 structures 385:394 complex oligosaccharide structures 361:394 complex oligosaccharide structures 361:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 1 3 theme eukaryotic 264:273 arg1 proteins 275:282 eukaryotic proteins 264:282 eukaryotic proteins 264:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 6 4 link N-linked 1101:1108 arg1 glycoproteins 1110:1122 N-linked glycoproteins 1101:1122 N-linked glycoproteins 1101:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 8 5 theme glycoproteins 1818:1830 arg1 properties 1804:1813 the conformational properties 1785:1813 the conformational properties of glycoproteins 1785:1830 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 6 theme low 1265:1267 arg1 temperature 1269:1279 low temperature 1265:1279 low temperature 1265:1279 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 7 7 link N-linked 1549:1556 arg1 glycan 1558:1563 the N-linked glycan 1545:1563 the N-linked glycan 1545:1563 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 1 8 theme frequent 188:195 arg1 glycosylation 162:174 Protein glycosylation 154:174 Protein glycosylation 154:174 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 8 theme frequent 188:195 arg1 modification 216:227 the most frequent post-translational modification 179:227 the most frequent post-translational modification 179:227 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 7 9 theme N-linked 1549:1556 arg1 glycan 1558:1563 the N-linked glycan 1545:1563 the N-linked glycan 1545:1563 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 7 10 theme native 1498:1503 arg1 dependent 1532:1540 dependent 1532:1540 dependent 1532:1540 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 7 10 theme native 1498:1503 arg1 dynamics 1520:1527 the native conformational dynamics 1494:1527 the native conformational dynamics 1494:1527 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 6 11 dep pH 1258:1259 arg1 i.e. 1245:1248 i.e. 1245:1248 i.e. 1245:1248 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 2 12 theme modifications 323:335 arg1 subset 313:318 a wide subset 306:318 a wide subset of modifications involving many types of complex oligosaccharide structures 306:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 7 13 theme conformational 1505:1518 arg1 dependent 1532:1540 dependent 1532:1540 dependent 1532:1540 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 7 13 theme conformational 1505:1518 arg1 dynamics 1520:1527 the native conformational dynamics 1494:1527 the native conformational dynamics 1494:1527 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 8 14 theme PNGase 1614:1619 arg1 deglycosylation 1623:1637 integrated PNGase A deglycosylation 1603:1637 integrated PNGase A deglycosylation 1603:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 5 15 link N-linked 850:857 arg1 glycan 859:864 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 0 16 theme Glycoproteins 139:151 arg1 Analysis 118:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis 72:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins 72:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 5 17 theme peptic 949:954 arg1 peptides 971:978 peptic N-glycosylated peptides 949:978 peptic N-glycosylated peptides that can be difficult to detect 949:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 6 18 theme novel 1032:1036 arg1 workflow 1045:1052 a novel HDX-MS workflow 1030:1052 a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature 1030:1279 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 8 19 link N-linked 1704:1711 arg1 sites 1720:1724 N-linked glycan sites 1704:1724 N-linked glycan sites 1704:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 3 20 theme protein 608:614 arg1 dynamics 631:638 protein conformational dynamics 608:638 protein conformational dynamics of complex heterogeneous proteins in solution 608:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 7 21 theme antibody 1440:1447 arg1 Trastuzumab 1449:1459 the monoclonal antibody Trastuzumab 1425:1459 the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan 1425:1563 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 5 22 theme N-linked 850:857 arg1 glycan 859:864 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 6 23 theme N-linked 1101:1108 arg1 glycoproteins 1110:1122 N-linked glycoproteins 1101:1122 N-linked glycoproteins 1101:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 8 24 theme HDX-MS 1582:1587 arg1 workflow 1589:1596 the HDX-MS workflow 1578:1596 the HDX-MS workflow with integrated PNGase A deglycosylation 1578:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 25 gly glycopeptides 1205:1217 arg2 glycopeptides 1205:1217 labeled peptic N-linked glycopeptides 1181:1217 labeled peptic N-linked glycopeptides 1181:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 6 26 theme dynamics 1089:1096 arg1 analysis 1058:1065 analysis 1058:1065 analysis of the conformational dynamics of N-linked glycoproteins 1058:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 0 27 theme Exchange 91:98 arg1 Analysis 118:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis 72:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins 72:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 3 28 theme conformational 616:629 arg1 dynamics 631:638 protein conformational dynamics 608:638 protein conformational dynamics of complex heterogeneous proteins in solution 608:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 0 29 from pH 45:46 arg1 Removal 0:6 Removal 0:6 Removal of N-Linked Glycosylations at Acidic pH by PNGase A 0:58 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 2 30 theme glycoproteins 427:439 arg1 analysis 415:422 structural analysis 404:422 structural analysis of glycoproteins and their glycans challenging for most analytical techniques 404:500 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 0 31 theme Spectrometry 105:116 arg1 Analysis 118:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis 72:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins 72:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 0 32 theme Acidic 38:43 arg1 pH 45:46 Acidic pH 38:46 Acidic pH 38:46 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 7 33 theme monoclonal 1429:1438 arg1 Trastuzumab 1449:1459 the monoclonal antibody Trastuzumab 1425:1459 the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan 1425:1563 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 8 34 with workflow 1589:1596 arg1 deglycosylation 1623:1637 integrated PNGase A deglycosylation 1603:1637 integrated PNGase A deglycosylation 1603:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 2 35 theme most 475:478 arg1 techniques 491:500 most analytical techniques 475:500 most analytical techniques 475:500 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 0 36 theme N-Linked 11:18 arg1 Glycosylations 20:33 N-Linked Glycosylations 11:33 N-Linked Glycosylations 11:33 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 6 37 theme PNGase 1149:1154 arg1 A 1156:1156 the enzyme PNGase A 1138:1156 the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides 1138:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 7 38 theme approach 1381:1388 arg1 utility 1365:1371 the utility 1361:1371 the utility of this approach 1361:1388 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 8 39 theme HDX 1670:1672 arg1 analysis 1647:1654 analysis 1647:1654 analysis of the native HDX of protein regions containing N-linked glycan sites 1647:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 0 40 theme PNGase 51:56 arg1 A 58:58 PNGase A 51:58 PNGase A 51:58 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 2 41 theme complex 361:367 arg1 structures 385:394 complex oligosaccharide structures 361:394 complex oligosaccharide structures 361:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 8 42 theme protein 1677:1683 arg1 regions 1685:1691 protein regions 1677:1691 protein regions containing N-linked glycan sites 1677:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 43 theme HDX 1222:1224 arg1 conditions 1233:1242 HDX quench conditions 1222:1242 HDX quench conditions 1222:1242 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 4 44 theme N-linked 687:694 arg1 glycoproteins 696:708 N-linked glycoproteins 687:708 N-linked glycoproteins 687:708 N-linked glycoproteins however pose a challenge for HDX-MS. 28193043 1 45 attach present 236:242 arg2 modification 216:227 the most frequent post-translational modification 179:227 the most frequent post-translational modification 179:227 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 45 attach present 236:242 arg2 glycosylation 162:174 Protein glycosylation 154:174 Protein glycosylation 154:174 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 45 attach present 236:242 arg1 % 259:259 more than 50% 247:259 more than 50% of eukaryotic proteins 247:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 45 attach present 236:242 arg1 proteins 275:282 eukaryotic proteins 264:282 eukaryotic proteins 264:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 5 46 theme peptides 971:978 arg1 diversity 936:944 a large diversity 928:944 a large diversity of peptic N-glycosylated peptides that can be difficult to detect 928:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 6 47 theme glycopeptides 1205:1217 arg1 deglycosylation 1162:1176 deglycosylation 1162:1176 deglycosylation of labeled peptic N-linked glycopeptides 1162:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 8 48 theme glycan 1713:1718 arg1 sites 1720:1724 N-linked glycan sites 1704:1724 N-linked glycan sites 1704:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 49 gly glycoproteins 1110:1122 arg1 glycoproteins 1110:1122 N-linked glycoproteins 1101:1122 N-linked glycoproteins 1101:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 2 50 theme structures 385:394 arg1 types 352:356 many types 347:356 many types of complex oligosaccharide structures 347:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 5 51 gly N-glycosylated 956:969 arg1 peptides 971:978 peptic N-glycosylated peptides 949:978 peptic N-glycosylated peptides that can be difficult to detect 949:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 0 52 attach Removal 0:6 arg2 Glycosylations 20:33 N-Linked Glycosylations 11:33 N-Linked Glycosylations 11:33 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 0 52 attach Removal 0:6 arg3 pH 45:46 Acidic pH 38:46 Acidic pH 38:46 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 2 53 theme wide 308:311 arg1 subset 313:318 a wide subset 306:318 a wide subset of modifications involving many types of complex oligosaccharide structures 306:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 8 54 theme conformational 1789:1802 arg1 properties 1804:1813 the conformational properties 1785:1813 the conformational properties of glycoproteins 1785:1830 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 1 55 theme proteins 275:282 arg1 % 259:259 more than 50% 247:259 more than 50% of eukaryotic proteins 247:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 55 theme proteins 275:282 arg1 proteins 275:282 eukaryotic proteins 264:282 eukaryotic proteins 264:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 5 56 theme N-glycosylated 956:969 arg1 peptides 971:978 peptic N-glycosylated peptides 949:978 peptic N-glycosylated peptides that can be difficult to detect 949:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 3 57 theme complex 643:649 arg1 proteins 665:672 complex heterogeneous proteins 643:672 complex heterogeneous proteins in solution 643:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 1 58 theme post-translational 197:214 arg1 glycosylation 162:174 Protein glycosylation 154:174 Protein glycosylation 154:174 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 58 theme post-translational 197:214 arg1 modification 216:227 the most frequent post-translational modification 179:227 the most frequent post-translational modification 179:227 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 6 59 theme acidic 1251:1256 arg1 pH 1258:1259 acidic pH 1251:1259 acidic pH 1251:1259 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 0 60 theme N-Linked 130:137 arg1 Glycoproteins 139:151 N-Linked Glycoproteins 130:151 N-Linked Glycoproteins 130:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 3 61 theme dynamics 631:638 arg1 investigation 591:603 investigation 591:603 investigation of protein conformational dynamics of complex heterogeneous proteins in solution 591:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 5 62 theme large 930:934 arg1 diversity 936:944 a large diversity 928:944 a large diversity of peptic N-glycosylated peptides that can be difficult to detect 928:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 3 63 theme heterogeneous 651:663 arg1 proteins 665:672 complex heterogeneous proteins 643:672 complex heterogeneous proteins in solution 643:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 8 64 gly glycoproteins 1818:1830 arg1 glycoproteins 1818:1830 glycoproteins 1818:1830 glycoproteins 1818:1830 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 65 theme HDX-MS 1038:1043 arg1 workflow 1045:1052 a novel HDX-MS workflow 1030:1052 a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature 1030:1279 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 5 66 theme glycoprotein 813:824 arg1 regions 798:804 regions 798:804 regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 798:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 1 67 gly glycosylation 162:174 arg2 proteins 275:282 eukaryotic proteins 264:282 eukaryotic proteins 264:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 67 gly glycosylation 162:174 arg2 % 259:259 more than 50% 247:259 more than 50% of eukaryotic proteins 247:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 3 68 theme proteins 665:672 arg1 dynamics 631:638 protein conformational dynamics 608:638 protein conformational dynamics of complex heterogeneous proteins in solution 608:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 3 69 from dynamics 631:638 arg1 solution 677:684 solution 677:684 solution 677:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 5 70 theme actual 843:848 arg1 glycan 859:864 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 1 71 located present 236:242 arg2 modification 216:227 the most frequent post-translational modification 179:227 the most frequent post-translational modification 179:227 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 71 located present 236:242 arg2 glycosylation 162:174 Protein glycosylation 154:174 Protein glycosylation 154:174 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 71 located present 236:242 arg1 % 259:259 more than 50% 247:259 more than 50% of eukaryotic proteins 247:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 71 located present 236:242 arg1 proteins 275:282 eukaryotic proteins 264:282 eukaryotic proteins 264:282 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 72 theme Protein 154:160 arg1 glycosylation 162:174 Protein glycosylation 154:174 Protein glycosylation 154:174 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 1 72 theme Protein 154:160 arg1 modification 216:227 the most frequent post-translational modification 179:227 the most frequent post-translational modification 179:227 Protein glycosylation is the most frequent post-translational modification and is present on more than 50% of eukaryotic proteins. 28193043 6 73 theme glycoproteins 1110:1122 arg1 dynamics 1089:1096 the conformational dynamics 1070:1096 the conformational dynamics of N-linked glycoproteins 1070:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 7 74 theme Trastuzumab 1449:1459 arg1 analysis 1413:1420 analysis 1413:1420 analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan 1413:1563 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 0 75 theme Hydrogen/Deuterium 72:89 arg1 Analysis 118:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis 72:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins 72:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 8 76 gly deglycosylation 1623:1637 arg1 workflow 1589:1596 the HDX-MS workflow 1578:1596 the HDX-MS workflow with integrated PNGase A deglycosylation 1578:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 8 77 theme integrated 1603:1612 arg1 deglycosylation 1623:1637 integrated PNGase A deglycosylation 1603:1637 integrated PNGase A deglycosylation 1603:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 0 78 theme Mass 100:103 arg1 Analysis 118:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis 72:125 Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins 72:151 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 4 79 gly glycoproteins 696:708 arg1 glycoproteins 696:708 N-linked glycoproteins 687:708 N-linked glycoproteins 687:708 N-linked glycoproteins however pose a challenge for HDX-MS. 28193043 6 80 theme conformational 1074:1087 arg1 dynamics 1089:1096 the conformational dynamics 1070:1096 the conformational dynamics of N-linked glycoproteins 1070:1122 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 7 81 theme A-based 1289:1295 arg1 deglycosylation 1297:1311 PNGase A-based deglycosylation 1282:1311 PNGase A-based deglycosylation 1282:1311 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 8 82 theme A 1621:1621 arg1 deglycosylation 1623:1637 integrated PNGase A deglycosylation 1603:1637 integrated PNGase A deglycosylation 1603:1637 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 2 83 theme many 347:350 arg1 types 352:356 many types 347:356 many types of complex oligosaccharide structures 347:394 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 7 84 theme PNGase 1282:1287 arg1 deglycosylation 1297:1311 PNGase A-based deglycosylation 1282:1311 PNGase A-based deglycosylation 1282:1311 PNGase A-based deglycosylation is thus performed after labeling (post-HDX) and the utility of this approach is demonstrated during analysis of the monoclonal antibody Trastuzumab for which it has been shown that the native conformational dynamics is dependent on the N-linked glycan. 28193043 6 85 theme labeled 1181:1187 arg1 glycopeptides 1205:1217 labeled peptic N-linked glycopeptides 1181:1217 labeled peptic N-linked glycopeptides 1181:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 2 86 gly glycoproteins 427:439 arg1 glycoproteins 427:439 glycoproteins 427:439 glycoproteins 427:439 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 5 87 theme pepsin 904:909 arg1 digestion 911:919 pepsin digestion 904:919 pepsin digestion 904:919 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 5 88 contain contain 831:837 arg1 glycoprotein 813:824 the glycoprotein 809:824 the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 809:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 5 88 contain contain 831:837 arg2 glycan 859:864 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 839:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 2 89 theme analytical 480:489 arg1 techniques 491:500 most analytical techniques 475:500 most analytical techniques 475:500 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 8 90 contain containing 1693:1702 arg2 sites 1720:1724 N-linked glycan sites 1704:1724 N-linked glycan sites 1704:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 8 90 contain containing 1693:1702 arg1 regions 1685:1691 protein regions 1677:1691 protein regions containing N-linked glycan sites 1677:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 5 91 gly glycoprotein 813:824 arg1 glycoprotein 813:824 the glycoprotein 809:824 the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect 809:1010 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 3 92 from proteins 665:672 arg1 solution 677:684 solution 677:684 solution 677:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 2 93 theme structural 404:413 arg1 analysis 415:422 structural analysis 404:422 structural analysis of glycoproteins and their glycans challenging for most analytical techniques 404:500 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 0 94 theme Glycosylations 20:33 arg1 Removal 0:6 Removal 0:6 Removal of N-Linked Glycosylations at Acidic pH by PNGase A 0:58 Removal of N-Linked Glycosylations at Acidic pH by PNGase A Facilitates Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of N-Linked Glycoproteins. 28193043 8 95 theme native 1663:1668 arg1 HDX 1670:1672 the native HDX 1659:1672 the native HDX of protein regions containing N-linked glycan sites 1659:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 3 96 from solution 677:684 arg1 dynamics 631:638 protein conformational dynamics 608:638 protein conformational dynamics of complex heterogeneous proteins in solution 608:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 2 97 theme glycans 451:457 arg1 analysis 415:422 structural analysis 404:422 structural analysis of glycoproteins and their glycans challenging for most analytical techniques 404:500 Glycosylation covers a wide subset of modifications involving many types of complex oligosaccharide structures, making structural analysis of glycoproteins and their glycans challenging for most analytical techniques. 28193043 5 98 theme glycan 869:874 arg1 heterogeneity 876:888 glycan heterogeneity 869:888 glycan heterogeneity combined with pepsin digestion 869:919 HDX information can typically not be obtained from regions of the glycoprotein that contain the actual N-linked glycan as glycan heterogeneity combined with pepsin digestion yields a large diversity of peptic N-glycosylated peptides that can be difficult to detect. 28193043 6 99 theme enzyme 1142:1147 arg1 A 1156:1156 the enzyme PNGase A 1138:1156 the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides 1138:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 3 100 theme Hydrogen/deuterium 503:520 arg1 technique 577:585 a sensitive technique 565:585 a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution 565:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 3 100 theme Hydrogen/deuterium 503:520 arg1 exchange 522:529 Hydrogen/deuterium exchange 503:529 Hydrogen/deuterium exchange monitored by mass spectrometry 503:560 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 4 101 link N-linked 687:694 arg1 glycoproteins 696:708 N-linked glycoproteins 687:708 N-linked glycoproteins 687:708 N-linked glycoproteins however pose a challenge for HDX-MS. 28193043 8 102 theme regions 1685:1691 arg1 HDX 1670:1672 the native HDX 1659:1672 the native HDX of protein regions containing N-linked glycan sites 1659:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 103 link N-linked 1196:1203 arg1 glycopeptides 1205:1217 labeled peptic N-linked glycopeptides 1181:1217 labeled peptic N-linked glycopeptides 1181:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 3 104 theme sensitive 567:575 arg1 exchange 522:529 Hydrogen/deuterium exchange 503:529 Hydrogen/deuterium exchange monitored by mass spectrometry 503:560 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 3 104 theme sensitive 567:575 arg1 technique 577:585 a sensitive technique 565:585 a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution 565:684 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 28193043 6 105 theme quench 1226:1231 arg1 conditions 1233:1242 HDX quench conditions 1222:1242 HDX quench conditions 1222:1242 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 8 106 theme N-linked 1704:1711 arg1 sites 1720:1724 N-linked glycan sites 1704:1724 N-linked glycan sites 1704:1724 In summary, the HDX-MS workflow with integrated PNGase A deglycosylation enables analysis of the native HDX of protein regions containing N-linked glycan sites and should thus significantly improve our ability to study the conformational properties of glycoproteins. 28193043 6 107 gly deglycosylation 1162:1176 arg1 glycopeptides 1205:1217 labeled peptic N-linked glycopeptides 1181:1217 labeled peptic N-linked glycopeptides 1181:1217 Here, we present a novel HDX-MS workflow for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. 28193043 3 108 theme mass 544:547 arg1 spectrometry 549:560 mass spectrometry 544:560 mass spectrometry 544:560 Hydrogen/deuterium exchange monitored by mass spectrometry is a sensitive technique for investigation of protein conformational dynamics of complex heterogeneous proteins in solution. 24747898 4 0 theme purified 863:870 arg1 variants 877:884 the purified CBH1 variants 859:884 the purified CBH1 variants 859:884 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 3 1 theme unfolded 689:696 arg1 genes 728:732 the unfolded protein response (UPR) target genes 685:732 the unfolded protein response (UPR) target genes 685:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 2 2 theme N-linked 361:368 arg1 glycans 370:376 the three N-linked glycans 351:376 the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 351:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 8 3 theme proper 1740:1745 arg1 interactions 1747:1758 tuning proper interactions 1733:1758 tuning proper interactions between glycans and functional residues 1733:1798 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 4 4 theme variants 877:884 arg1 characterization 839:854 Further characterization 831:854 Further characterization of the purified CBH1 variants 831:884 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 2 5 from significance 335:346 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 5 from significance 335:346 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 6 theme structure 1465:1473 arg1 integrity 1434:1442 the integrity 1430:1442 the integrity of regular secondary structure 1430:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 4 7 theme catalyzed 1045:1053 arg1 hydrolysis 1055:1064 the catalyzed hydrolysis 1041:1064 the catalyzed hydrolysis of soluble substrate 1041:1085 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 0 8 from reesei 144:149 arg1 secretion 69:77 secretion 69:77 secretion 69:77 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 8 from reesei 144:149 arg1 activity 80:87 activity 80:87 activity 80:87 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 8 from reesei 144:149 arg1 stability 94:102 stability 94:102 stability 94:102 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 3 9 theme target 721:726 arg1 genes 728:732 the unfolded protein response (UPR) target genes 685:732 the unfolded protein response (UPR) target genes 685:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 10 theme N-glycosylation 542:556 arg1 sites 558:562 one or two N-glycosylation sites 531:562 one or two N-glycosylation sites 531:562 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 4 11 theme Further 831:837 arg1 characterization 839:854 Further characterization 831:854 Further characterization of the purified CBH1 variants 831:884 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 7 12 theme regular 1447:1453 arg1 structure 1465:1473 regular secondary structure 1447:1473 regular secondary structure 1447:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 3 13 theme UPR 716:718 arg1 genes 728:732 the unfolded protein response (UPR) target genes 685:732 the unfolded protein response (UPR) target genes 685:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 1 14 from effects 294:300 arg1 proteins 305:312 proteins 305:312 proteins 305:312 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 2 15 from secretion 486:494 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 15 from secretion 486:494 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 16 theme pronounced 1409:1418 arg1 effect 1420:1425 a more pronounced effect 1402:1425 a more pronounced effect on the integrity of regular secondary structure 1402:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 3 17 theme variant 762:768 arg1 secretion 739:747 secretion 739:747 secretion of this CBH1 variant 739:768 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 18 theme CBH1 611:614 arg1 secretion 598:606 the extracellular secretion 580:606 the extracellular secretion of CBH1 580:614 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 2 19 from activity 500:507 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 19 from activity 500:507 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 0 20 theme Trichoderma 132:142 arg1 reesei 144:149 Trichoderma reesei 132:149 Trichoderma reesei 132:149 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 7 21 theme N-glycosylation 1372:1386 arg1 removal 1361:1367 removal 1361:1367 removal of N-glycosylation at Asn384 1361:1396 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 3 22 gly N-glycosylation 542:556 arg2 one 531:533 one 531:533 one 531:533 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 22 gly N-glycosylation 542:556 arg2 sites 558:562 one or two N-glycosylation sites 531:562 one or two N-glycosylation sites 531:562 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 22 gly N-glycosylation 542:556 arg2 two 538:540 two 538:540 two 538:540 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 6 23 from severe 1248:1253 arg1 contrast 1200:1207 contrast 1200:1207 contrast 1200:1207 In contrast, this thermal labile property was less severe when hydrolyzing insoluble cellulose. 24747898 3 24 gly glycosylation 640:652 arg2 sites 654:658 the glycosylation sites 636:658 the glycosylation sites 636:658 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 0 25 from effect 16:21 arg1 secretion 69:77 secretion 69:77 secretion 69:77 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 25 from effect 16:21 arg1 activity 80:87 activity 80:87 activity 80:87 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 25 from effect 16:21 arg1 stability 94:102 stability 94:102 stability 94:102 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 3 26 theme deletion 814:821 arg1 strain 823:828 a calnexin gene deletion strain 798:828 a calnexin gene deletion strain 798:828 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 5 27 theme glycans 1129:1135 arg1 loss 1102:1105 Combinatorial loss 1088:1105 Combinatorial loss of these two N-linked glycans 1088:1135 Combinatorial loss of these two N-linked glycans further exacerbated the temperature-dependent inactivation. 24747898 1 28 theme N-linked 152:159 arg1 glycosylation 161:173 N-linked glycosylation 152:173 N-linked glycosylation 152:173 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 3 29 theme calnexin 800:807 arg1 strain 823:828 a calnexin gene deletion strain 798:828 a calnexin gene deletion strain 798:828 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 5 30 theme Combinatorial 1088:1100 arg1 loss 1102:1105 Combinatorial loss 1088:1105 Combinatorial loss of these two N-linked glycans 1088:1135 Combinatorial loss of these two N-linked glycans further exacerbated the temperature-dependent inactivation. 24747898 4 31 theme CBH1 945:948 arg1 reactivity 931:940 the thermal reactivity 919:940 the thermal reactivity of CBH1 919:948 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 2 32 theme cellobiohydrolase 405:421 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 32 theme cellobiohydrolase 405:421 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 33 from Asn384 1391:1396 arg1 removal 1361:1367 removal 1361:1367 removal of N-glycosylation at Asn384 1361:1396 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 4 34 theme Asn384 1008:1013 arg1 site 1029:1032 Asn384 glycosylation site 1008:1032 Asn384 glycosylation site 1008:1032 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 8 35 theme N-glycosylation 1559:1573 arg1 modifications 1575:1587 N-glycosylation modifications 1559:1587 N-glycosylation modifications 1559:1587 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 6 36 from contrast 1200:1207 arg1 severe 1248:1253 severe 1248:1253 severe 1248:1253 In contrast, this thermal labile property was less severe when hydrolyzing insoluble cellulose. 24747898 1 37 theme extrinsic 284:292 arg1 effects 294:300 both intrinsic and extrinsic effects 265:300 both intrinsic and extrinsic effects on proteins 265:312 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 4 38 theme site 1029:1032 arg1 removal 981:987 removal 981:987 removal of either Asn45 or Asn384 glycosylation site 981:1032 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 6 39 theme thermal 1215:1221 arg1 property 1230:1237 this thermal labile property 1210:1237 this thermal labile property 1210:1237 In contrast, this thermal labile property was less severe when hydrolyzing insoluble cellulose. 24747898 7 40 from effect 1420:1425 arg1 integrity 1434:1442 the integrity 1430:1442 the integrity of regular secondary structure 1430:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 7 41 theme variants 1338:1345 arg1 integrity 1320:1328 the structural integrity 1305:1328 the structural integrity of CBH1 variants 1305:1345 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 2 42 from domain 395:400 arg1 activity 500:507 activity 500:507 activity 500:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 42 from domain 395:400 arg1 reesei 472:477 the filamentous fungus Trichoderma reesei 437:477 the filamentous fungus Trichoderma reesei 437:477 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 42 from domain 395:400 arg1 significance 335:346 the significance 331:346 the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 331:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 42 from domain 395:400 arg1 secretion 486:494 its secretion 482:494 its secretion 482:494 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 4 43 theme Asn45 999:1003 arg1 removal 981:987 removal 981:987 removal of either Asn45 or Asn384 glycosylation site 981:1032 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 3 44 theme glycosylation 640:652 arg1 sites 654:658 the glycosylation sites 636:658 the glycosylation sites 636:658 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 0 45 theme N-glycosylation 40:54 arg1 sites 56:60 the different N-glycosylation sites 26:60 the different N-glycosylation sites 26:60 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 5 46 theme temperature-dependent 1161:1181 arg1 inactivation 1183:1194 the temperature-dependent inactivation 1157:1194 the temperature-dependent inactivation 1157:1194 Combinatorial loss of these two N-linked glycans further exacerbated the temperature-dependent inactivation. 24747898 2 47 from cellobiohydrolase 405:421 arg1 activity 500:507 activity 500:507 activity 500:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 47 from cellobiohydrolase 405:421 arg1 secretion 486:494 its secretion 482:494 its secretion 482:494 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 48 theme structural 1309:1318 arg1 integrity 1320:1328 the structural integrity 1305:1328 the structural integrity of CBH1 variants 1305:1345 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 4 49 theme soluble 1069:1075 arg1 substrate 1077:1085 soluble substrate 1069:1085 soluble substrate 1069:1085 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 8 50 theme specific 1625:1632 arg1 regions 1645:1651 specific functional regions 1625:1651 specific functional regions of CBH1 1625:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 7 51 attach removal 1361:1367 arg2 N-glycosylation 1372:1386 N-glycosylation 1372:1386 N-glycosylation 1372:1386 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 7 51 attach removal 1361:1367 arg3 Asn384 1391:1396 Asn384 1391:1396 Asn384 1391:1396 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 8 52 theme regions 1645:1651 arg1 stability 1612:1620 the stability 1608:1620 the stability of specific functional regions of CBH1 1608:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 7 53 contain had 1398:1400 arg2 effect 1420:1425 a more pronounced effect 1402:1425 a more pronounced effect on the integrity of regular secondary structure 1402:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 7 53 contain had 1398:1400 arg1 removal 1361:1367 removal 1361:1367 removal of N-glycosylation at Asn384 1361:1396 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 7 54 theme Asn270 1508:1513 arg1 loss 1491:1494 the loss 1487:1494 the loss of Asn45 or Asn270 1487:1513 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 8 55 theme CBH1 1656:1659 arg1 regions 1645:1651 specific functional regions 1625:1651 specific functional regions of CBH1 1625:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 2 56 theme glycans 370:376 arg1 significance 335:346 the significance 331:346 the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 331:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 8 57 theme CBH1 1725:1728 arg1 thermostability 1706:1720 the thermostability 1702:1720 the thermostability of CBH1 1702:1728 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 7 58 theme Asn45 1499:1503 arg1 loss 1491:1494 the loss 1487:1494 the loss of Asn45 or Asn270 1487:1513 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 3 59 theme protein 698:704 arg1 genes 728:732 the unfolded protein response (UPR) target genes 685:732 the unfolded protein response (UPR) target genes 685:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 8 60 theme tuning 1733:1738 arg1 interactions 1747:1758 tuning proper interactions 1733:1758 tuning proper interactions between glycans and functional residues 1733:1798 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 1 61 theme intrinsic 270:278 arg1 effects 294:300 both intrinsic and extrinsic effects 265:300 both intrinsic and extrinsic effects on proteins 265:312 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 4 62 theme CBH1 872:875 arg1 variants 877:884 the purified CBH1 variants 859:884 the purified CBH1 variants 859:884 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 2 63 link N-linked 361:368 arg1 glycans 370:376 the three N-linked glycans 351:376 the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 351:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 64 from glycans 370:376 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 64 from glycans 370:376 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 65 theme secondary 1455:1463 arg1 structure 1465:1473 regular secondary structure 1447:1473 regular secondary structure 1447:1473 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 6 66 theme insoluble 1272:1280 arg1 cellulose 1282:1290 insoluble cellulose 1272:1290 insoluble cellulose 1272:1290 In contrast, this thermal labile property was less severe when hydrolyzing insoluble cellulose. 24747898 0 67 theme cellobiohydrolase 107:123 arg1 secretion 69:77 secretion 69:77 secretion 69:77 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 67 theme cellobiohydrolase 107:123 arg1 activity 80:87 activity 80:87 activity 80:87 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 0 67 theme cellobiohydrolase 107:123 arg1 stability 94:102 stability 94:102 stability 94:102 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 3 68 theme sites 558:562 arg1 removal 520:526 the removal 516:526 the removal of one or two N-glycosylation sites 516:562 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 8 69 theme functional 1780:1789 arg1 residues 1791:1798 functional residues 1780:1798 functional residues 1780:1798 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 3 70 theme extracellular 584:596 arg1 secretion 598:606 the extracellular secretion 580:606 the extracellular secretion of CBH1 580:614 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 71 theme CBH1 757:760 arg1 variant 762:768 this CBH1 variant 752:768 this CBH1 variant 752:768 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 3 72 theme genes 728:732 arg1 expression 671:680 expression 671:680 expression of the unfolded protein response (UPR) target genes 671:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 6 73 theme labile 1223:1228 arg1 property 1230:1237 this thermal labile property 1210:1237 this thermal labile property 1210:1237 In contrast, this thermal labile property was less severe when hydrolyzing insoluble cellulose. 24747898 5 74 theme N-linked 1120:1127 arg1 glycans 1129:1135 these two N-linked glycans 1110:1135 these two N-linked glycans 1110:1135 Combinatorial loss of these two N-linked glycans further exacerbated the temperature-dependent inactivation. 24747898 8 75 theme differential 1537:1548 arg1 roles 1550:1554 differential roles 1537:1554 differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 1537:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 4 76 theme thermal 923:929 arg1 reactivity 931:940 the thermal reactivity 919:940 the thermal reactivity of CBH1 919:948 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 2 77 from reesei 472:477 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 77 from reesei 472:477 arg1 CBH1 426:429 CBH1 426:429 CBH1 426:429 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 77 from reesei 472:477 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 3 78 theme gene 809:812 arg1 strain 823:828 a calnexin gene deletion strain 798:828 a calnexin gene deletion strain 798:828 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 8 79 theme modifications 1575:1587 arg1 roles 1550:1554 differential roles 1537:1554 differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 1537:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 2 80 theme catalytic 385:393 arg1 cellobiohydrolase 405:421 cellobiohydrolase I 405:423 cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 405:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 2 80 theme catalytic 385:393 arg1 domain 395:400 the catalytic domain 381:400 the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity 381:507 We investigated the significance of the three N-linked glycans on the catalytic domain of cellobiohydrolase I (CBH1) from the filamentous fungus Trichoderma reesei in its secretion and activity. 24747898 7 81 theme CBH1 1333:1336 arg1 variants 1338:1345 CBH1 variants 1333:1345 CBH1 variants 1333:1345 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 4 82 theme glycosylation 1015:1027 arg1 site 1029:1032 Asn384 glycosylation site 1008:1032 Asn384 glycosylation site 1008:1032 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 1 83 link N-linked 152:159 arg1 glycosylation 161:173 N-linked glycosylation 152:173 N-linked glycosylation 152:173 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 0 84 theme different 30:38 arg1 sites 56:60 the different N-glycosylation sites 26:60 the different N-glycosylation sites 26:60 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 5 85 link N-linked 1120:1127 arg1 glycans 1129:1135 these two N-linked glycans 1110:1135 these two N-linked glycans 1110:1135 Combinatorial loss of these two N-linked glycans further exacerbated the temperature-dependent inactivation. 24747898 7 86 theme integrity 1320:1328 arg1 Analysis 1293:1300 Analysis 1293:1300 Analysis of the structural integrity of CBH1 variants 1293:1345 Analysis of the structural integrity of CBH1 variants revealed that removal of N-glycosylation at Asn384 had a more pronounced effect on the integrity of regular secondary structure compared to the loss of Asn45 or Asn270. 24747898 1 87 theme eukaryotic 237:246 arg1 proteome 248:255 the eukaryotic proteome 233:255 the eukaryotic proteome 233:255 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 0 88 theme sites 56:60 arg1 effect 16:21 the effect 12:21 the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei 12:149 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 4 89 gly glycosylation 1015:1027 arg2 site 1029:1032 Asn384 glycosylation site 1008:1032 Asn384 glycosylation site 1008:1032 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 8 90 theme functional 1634:1643 arg1 regions 1645:1651 specific functional regions 1625:1651 specific functional regions of CBH1 1625:1659 These data implicate differential roles of N-glycosylation modifications in contributing to the stability of specific functional regions of CBH1 and highlight the potential of improving the thermostability of CBH1 by tuning proper interactions between glycans and functional residues. 24747898 0 91 gly N-glycosylation 40:54 arg2 sites 56:60 the different N-glycosylation sites 26:60 the different N-glycosylation sites 26:60 Deciphering the effect of the different N-glycosylation sites on the secretion, activity, and stability of cellobiohydrolase I from Trichoderma reesei. 24747898 4 92 theme substrate 1077:1085 arg1 hydrolysis 1055:1064 the catalyzed hydrolysis 1041:1064 the catalyzed hydrolysis of soluble substrate 1041:1085 Further characterization of the purified CBH1 variants showed that, compared to Asn270, the thermal reactivity of CBH1 was significantly decreased by removal of either Asn45 or Asn384 glycosylation site during the catalyzed hydrolysis of soluble substrate. 24747898 3 93 theme response 706:713 arg1 genes 728:732 the unfolded protein response (UPR) target genes 685:732 the unfolded protein response (UPR) target genes 685:732 While the removal of one or two N-glycosylation sites hardly affected the extracellular secretion of CBH1, eliminating all of the glycosylation sites did induce expression of the unfolded protein response (UPR) target genes, and secretion of this CBH1 variant was severely compromised in a calnexin gene deletion strain. 24747898 1 94 dep structures 205:214 arg1 the 201:203 the 201:203 the 201:203 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 1 95 theme proteome 248:255 arg1 structures 205:214 structures 205:214 structures 205:214 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 24747898 1 95 theme proteome 248:255 arg1 functions 220:228 functions 220:228 functions 220:228 N-linked glycosylation modulates and diversifies the structures and functions of the eukaryotic proteome through both intrinsic and extrinsic effects on proteins. 28433761 4 0 theme oligosaccharides 605:620 arg1 presence 593:600 the presence 589:600 the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid 589:681 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 6 1 attach present 1092:1098 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 6 1 attach present 1092:1098 arg2 sequons 1064:1070 ten characteristic sequons 1045:1070 ten characteristic sequons (Asn-X-Ser/Thr) 1045:1086 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 5 2 theme Further 684:690 arg1 analysis 705:712 Further nanoHPLC-ESI analysis 684:712 Further nanoHPLC-ESI analysis of the glycopeptides 684:733 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 2 3 theme S-layer 322:328 arg1 protein 330:336 the S-layer protein 318:336 the S-layer protein of Lactobacillus kefiri CIDCA 83111 318:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 9 4 theme structure 1483:1491 arg1 description 1464:1474 the first description 1454:1474 the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species 1454:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 9 4 theme structure 1483:1491 arg1 this 1446:1449 this 1446:1449 this 1446:1449 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 2 5 theme work 262:265 arg1 aim 250:252 The aim 246:252 The aim of this work 246:265 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 9 6 theme N-glycans 1496:1504 arg1 structure 1483:1491 the structure 1479:1491 the structure of N-glycans 1479:1504 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 5 7 dep motif 850:854 arg1 substituted 872:882 substituted 872:882 substituted on average with eight glucose residues 872:921 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 5 7 dep motif 850:854 arg1 decorated 927:935 decorated 927:935 decorated 927:935 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 5 8 theme glycosylation 836:848 arg1 motif 850:854 a signature glycosylation motif 824:854 a signature glycosylation motif 824:854 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 11 9 theme L. 1772:1773 arg1 strains 1782:1788 kefir-isolated L. kefiri strains 1757:1788 kefir-isolated L. kefiri strains 1757:1788 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 1 10 theme acid 168:171 arg1 bacteria 173:180 lactic acid bacteria 161:180 lactic acid bacteria 161:180 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 12 11 theme important 2075:2083 arg1 properties 2085:2094 these important properties 2069:2094 these important properties 2069:2094 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 5 12 gly glycopeptides 721:733 arg2 glycopeptides 721:733 the glycopeptides 717:733 the glycopeptides 717:733 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 11 13 from present 1853:1859 arg1 grains 1870:1875 kefir grains 1864:1875 kefir grains 1864:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 8 14 theme ion 1326:1328 arg1 modes 1330:1334 the positive and negative ion modes 1300:1334 the positive and negative ion modes 1300:1334 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 11 15 theme S-layer 1735:1741 arg1 proteins 1743:1750 S-layer proteins 1735:1750 S-layer proteins from kefir-isolated L. kefiri strains 1735:1788 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 5 16 gly O-glycosylated 972:985 arg1 site 987:990 another O-glycosylated site 964:990 another O-glycosylated site on peptide 471-476 964:1009 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 5 17 theme O-glycosylated 746:759 arg1 peptides 761:768 two O-glycosylated peptides 742:768 two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure 742:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 1 18 theme glycosylation 191:203 arg1 systems 205:211 general glycosylation systems 183:211 general glycosylation systems 183:211 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 13 19 from spp 2206:2208 arg1 this 2110:2113 this 2110:2113 this 2110:2113 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 13 19 from spp 2206:2208 arg1 description 2128:2138 the first description 2118:2138 the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp 2118:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 13 19 from spp 2206:2208 arg1 glycoprotein 2174:2185 S-layer glycoprotein 2166:2185 S-layer glycoprotein from Lactobacillus spp 2166:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 10 20 dep SIGNIFICANCE 1559:1570 arg1 characterization 1583:1598 A detailed characterization 1572:1598 SIGNIFICANCE A detailed characterization of protein glycosylation 1559:1623 SIGNIFICANCE A detailed characterization of protein glycosylation is essential to establish the basis for understanding and investigating its biological role. 28433761 9 21 gly glycoproteins 1517:1529 arg1 glycoproteins 1517:1529 S-layer glycoproteins 1509:1529 S-layer glycoproteins from Lactobacillus species 1509:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 5 22 gly O-glycosylated 746:759 arg1 peptides 761:768 two O-glycosylated peptides 742:768 two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure 742:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 11 23 theme adverse 1916:1922 arg1 effects 1924:1930 the adverse effects 1912:1930 the adverse effects of different enteric pathogens 1912:1961 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 5 24 theme O-glycosylated 972:985 arg1 site 987:990 another O-glycosylated site 964:990 another O-glycosylated site on peptide 471-476 964:1009 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 1 25 theme Gram-positive 129:141 arg1 bacteria 173:180 lactic acid bacteria 161:180 lactic acid bacteria 161:180 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 1 25 theme Gram-positive 129:141 arg1 bacteria 143:150 Gram-positive bacteria 129:150 Gram-positive bacteria 129:150 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 5 26 dep peptides 761:768 arg1 SSASSASSA 792:800 the peptide sequence SSASSASSA 771:800 two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure 742:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 13 27 from description 2128:2138 arg1 glycoprotein 2174:2185 S-layer glycoprotein 2166:2185 S-layer glycoprotein from Lactobacillus spp 2166:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 13 27 from description 2128:2138 arg1 spp 2206:2208 Lactobacillus spp 2192:2208 Lactobacillus spp 2192:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 5 28 from motif 850:854 arg1 buchneri 862:869 L. buchneri 859:869 L. buchneri 859:869 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 11 29 attach present 1853:1859 arg2 yeasts 1846:1851 yeasts 1846:1851 yeasts present in kefir grains 1846:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 11 29 attach present 1853:1859 arg1 grains 1870:1875 kefir grains 1864:1875 kefir grains 1864:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 5 30 theme galacturonic 942:953 arg1 acid 955:958 galacturonic acid 942:958 galacturonic acid 942:958 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 12 31 from chains 2023:2028 arg1 protein 2038:2044 this protein 2033:2044 this protein 2033:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 8 32 theme N-glycan 1407:1414 arg1 structures 1416:1425 short N-glycan structures 1401:1425 short N-glycan structures 1401:1425 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 0 33 theme kefiri 81:86 arg1 83111 94:98 Lactobacillus kefiri CIDCA 83111 67:98 Lactobacillus kefiri CIDCA 83111 67:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 13 34 theme Lactobacillus 2192:2204 arg1 spp 2206:2208 Lactobacillus spp 2192:2208 Lactobacillus spp 2192:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 12 35 theme chains 2023:2028 arg1 site 2004:2007 site 2004:2007 site 2004:2007 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 12 35 theme chains 2023:2028 arg1 chains 2023:2028 glycosidic chains 2012:2028 glycosidic chains in this protein 2012:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 12 35 theme chains 2023:2028 arg1 type 1995:1998 type 1995:1998 type 1995:1998 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 5 36 from acid 955:958 arg1 471-476 1003:1009 peptide 471-476 995:1009 peptide 471-476 995:1009 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 6 37 theme characteristic 1049:1062 arg1 sequons 1064:1070 ten characteristic sequons 1045:1070 ten characteristic sequons (Asn-X-Ser/Thr) 1045:1086 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 0 38 theme 83111 94:98 arg1 O- 103:104 O- 103:104 O- 103:104 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 0 38 theme 83111 94:98 arg1 glycoprotein 51:62 the S-layer glycoprotein 39:62 the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 39:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 13 39 theme N-glycosidic 2143:2154 arg1 chains 2156:2161 N-glycosidic chains 2143:2161 N-glycosidic chains 2143:2161 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 5 40 theme Glc5-8GalA2 1019:1029 arg1 structure 1031:1039 a Glc5-8GalA2 structure 1017:1039 a Glc5-8GalA2 structure 1017:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 2 41 dep Lactobacillus 341:353 arg1 kefiri 355:360 kefiri 355:360 kefiri 355:360 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 3 42 theme anion 423:427 arg1 chromatography 462:475 anion exchange high performance liquid chromatography 423:475 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 7 43 theme short 1261:1265 arg1 chains 1276:1281 mainly short N-linked chains 1254:1281 mainly short N-linked chains 1254:1281 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 5 44 from site 987:990 arg1 471-476 1003:1009 peptide 471-476 995:1009 peptide 471-476 995:1009 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 2 45 theme CIDCA 362:366 arg1 83111 368:372 Lactobacillus kefiri CIDCA 83111 341:372 Lactobacillus kefiri CIDCA 83111 341:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 6 46 theme amino 1115:1119 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 11 47 from strains 1782:1788 arg1 proteins 1743:1750 S-layer proteins 1735:1750 S-layer proteins from kefir-isolated L. kefiri strains 1735:1788 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 3 48 theme liquid 455:460 arg1 chromatography 462:475 anion exchange high performance liquid chromatography 423:475 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 7 49 theme chromatography 1223:1236 arg1 analysis 1238:1245 Anion exchange chromatography analysis 1208:1245 Anion exchange chromatography analysis 1208:1245 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 3 50 theme β-elimination 387:399 arg1 useful 490:495 useful 490:495 useful 490:495 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 3 50 theme β-elimination 387:399 arg1 treatment 401:409 A reductive β-elimination treatment 375:409 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis 375:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 10 51 theme detailed 1574:1581 arg1 characterization 1583:1598 A detailed characterization 1572:1598 SIGNIFICANCE A detailed characterization of protein glycosylation 1559:1623 SIGNIFICANCE A detailed characterization of protein glycosylation is essential to establish the basis for understanding and investigating its biological role. 28433761 3 52 theme high 438:441 arg1 chromatography 462:475 anion exchange high performance liquid chromatography 423:475 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 1 53 theme lactic 161:166 arg1 bacteria 173:180 lactic acid bacteria 161:180 lactic acid bacteria 161:180 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 11 54 dep L. 1772:1773 arg1 kefiri 1775:1780 kefiri 1775:1780 kefiri 1775:1780 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 7 55 theme Anion 1208:1212 arg1 chromatography 1223:1236 Anion exchange chromatography 1208:1236 Anion exchange chromatography analysis 1208:1245 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 4 56 contain carrying 656:663 arg1 units 650:654 5 to 8 glucose units 635:654 5 to 8 glucose units carrying galacturonic acid 635:681 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 4 56 contain carrying 656:663 arg2 acid 678:681 galacturonic acid 665:681 galacturonic acid 665:681 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 13 57 theme first 2122:2126 arg1 this 2110:2113 this 2110:2113 this 2110:2113 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 13 57 theme first 2122:2126 arg1 description 2128:2138 the first description 2118:2138 the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp 2118:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 0 58 theme S-layer 43:49 arg1 O- 103:104 O- 103:104 O- 103:104 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 0 58 theme S-layer 43:49 arg1 glycoprotein 51:62 the S-layer glycoprotein 39:62 the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 39:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 4 59 theme mass 552:555 arg1 analysis 570:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 10 60 theme glycosylation 1611:1623 arg1 characterization 1583:1598 A detailed characterization 1572:1598 SIGNIFICANCE A detailed characterization of protein glycosylation 1559:1623 SIGNIFICANCE A detailed characterization of protein glycosylation is essential to establish the basis for understanding and investigating its biological role. 28433761 5 61 gly glycosylation 836:848 arg2 motif 850:854 a signature glycosylation motif 824:854 a signature glycosylation motif 824:854 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 11 62 theme enteric 1945:1951 arg1 pathogens 1953:1961 different enteric pathogens 1935:1961 different enteric pathogens 1935:1961 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 6 63 theme F 1158:1158 arg1 digestion 1160:1168 a PNGase F digestion 1149:1168 a PNGase F digestion 1149:1168 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 11 64 theme bacterial 1825:1833 arg1 cells 1835:1839 bacterial cells 1825:1839 bacterial cells 1825:1839 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 3 65 theme O-glycosidic 517:528 arg1 structures 530:539 the O-glycosidic structures 513:539 the O-glycosidic structures 513:539 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 9 66 theme S-layer 1509:1515 arg1 glycoproteins 1517:1529 S-layer glycoproteins 1509:1529 S-layer glycoproteins from Lactobacillus species 1509:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 0 67 gly glycoprotein 51:62 arg1 O- 103:104 O- 103:104 O- 103:104 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 0 67 gly glycoprotein 51:62 arg1 glycoprotein 51:62 the S-layer glycoprotein 39:62 the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 39:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 11 68 theme kefir-isolated 1757:1770 arg1 strains 1782:1788 kefir-isolated L. kefiri strains 1757:1788 kefir-isolated L. kefiri strains 1757:1788 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 2 69 theme protein 330:336 arg1 glycosylation 301:313 the glycosylation 297:313 the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111 297:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 5 70 theme nanoHPLC-ESI 692:703 arg1 analysis 705:712 Further nanoHPLC-ESI analysis 684:712 Further nanoHPLC-ESI analysis of the glycopeptides 684:733 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 6 71 from sequence 1126:1133 arg1 present 1092:1098 present 1092:1098 present 1092:1098 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 7 72 link N-linked 1267:1274 arg1 chains 1276:1281 mainly short N-linked chains 1254:1281 mainly short N-linked chains 1254:1281 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 5 73 theme glucose 906:912 arg1 residues 914:921 eight glucose residues 900:921 eight glucose residues 900:921 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 8 74 from NanoHPLC-ESI 1284:1295 arg1 modes 1330:1334 the positive and negative ion modes 1300:1334 the positive and negative ion modes 1300:1334 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 8 75 theme positive 1304:1311 arg1 modes 1330:1334 the positive and negative ion modes 1300:1334 the positive and negative ion modes 1300:1334 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 6 76 from present 1092:1098 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 6 77 dep sequons 1064:1070 arg1 Asn-X-Ser/Thr 1073:1085 Asn-X-Ser/Thr 1073:1085 Asn-X-Ser/Thr 1073:1085 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 12 78 from type 1995:1998 arg1 protein 2038:2044 this protein 2033:2044 this protein 2033:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 8 79 theme negative 1317:1324 arg1 modes 1330:1334 the positive and negative ion modes 1300:1334 the positive and negative ion modes 1300:1334 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 10 80 theme biological 1701:1710 arg1 role 1712:1715 its biological role 1697:1715 its biological role 1697:1715 SIGNIFICANCE A detailed characterization of protein glycosylation is essential to establish the basis for understanding and investigating its biological role. 28433761 1 81 theme general 183:189 arg1 systems 205:211 general glycosylation systems 183:211 general glycosylation systems 183:211 In Gram-positive bacteria, such as lactic acid bacteria, general glycosylation systems have not been documented so far. 28433761 2 82 theme Lactobacillus 341:353 arg1 83111 368:372 Lactobacillus kefiri CIDCA 83111 341:372 Lactobacillus kefiri CIDCA 83111 341:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 6 83 theme N-linked 1181:1188 arg1 oligosaccharides 1190:1205 N-linked oligosaccharides 1181:1205 N-linked oligosaccharides 1181:1205 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 11 84 with interaction 1810:1820 arg1 yeasts 1846:1851 yeasts 1846:1851 yeasts present in kefir grains 1846:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 4 85 theme galacturonic 665:676 arg1 acid 678:681 galacturonic acid 665:681 galacturonic acid 665:681 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 11 86 theme different 1935:1943 arg1 pathogens 1953:1961 different enteric pathogens 1935:1961 different enteric pathogens 1935:1961 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 11 87 from grains 1870:1875 arg1 present 1853:1859 present 1853:1859 present 1853:1859 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 13 88 gly glycoprotein 2174:2185 arg1 glycoprotein 2174:2185 S-layer glycoprotein 2166:2185 S-layer glycoprotein from Lactobacillus spp 2166:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 5 89 theme glycopeptides 721:733 arg1 analysis 705:712 Further nanoHPLC-ESI analysis 684:712 Further nanoHPLC-ESI analysis of the glycopeptides 684:733 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 4 90 dep 8 640:640 arg1 to 637:638 to 637:638 to 637:638 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 12 91 from protein 2038:2044 arg1 site 2004:2007 site 2004:2007 site 2004:2007 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 12 91 from protein 2038:2044 arg1 chains 2023:2028 glycosidic chains 2012:2028 glycosidic chains in this protein 2012:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 12 91 from protein 2038:2044 arg1 type 1995:1998 type 1995:1998 type 1995:1998 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 8 92 theme different 1365:1373 arg1 peptides 1375:1382 two different peptides 1361:1382 two different peptides substituted with short N-glycan structures 1361:1425 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 11 93 theme kefir 1864:1868 arg1 grains 1870:1875 kefir grains 1864:1875 kefir grains 1864:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 0 94 theme Lactobacillus 67:79 arg1 83111 94:98 Lactobacillus kefiri CIDCA 83111 67:98 Lactobacillus kefiri CIDCA 83111 67:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 11 95 theme present 1853:1859 arg1 yeasts 1846:1851 yeasts 1846:1851 yeasts present in kefir grains 1846:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 9 96 theme Lactobacillus 1536:1548 arg1 species 1550:1556 Lactobacillus species 1536:1556 Lactobacillus species 1536:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 5 97 theme peptide 775:781 arg1 SSASSASSA 792:800 the peptide sequence SSASSASSA 771:800 two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure 742:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 0 98 theme CIDCA 88:92 arg1 83111 94:98 Lactobacillus kefiri CIDCA 83111 67:98 Lactobacillus kefiri CIDCA 83111 67:98 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 10 99 theme protein 1603:1609 arg1 glycosylation 1611:1623 protein glycosylation 1603:1623 protein glycosylation 1603:1623 SIGNIFICANCE A detailed characterization of protein glycosylation is essential to establish the basis for understanding and investigating its biological role. 28433761 6 100 located present 1092:1098 arg2 sequons 1064:1070 ten characteristic sequons 1045:1070 ten characteristic sequons (Asn-X-Ser/Thr) 1045:1086 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 6 100 located present 1092:1098 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 8 101 theme short 1401:1405 arg1 structures 1416:1425 short N-glycan structures 1401:1425 short N-glycan structures 1401:1425 NanoHPLC-ESI in the positive and negative ion modes were useful to determine two different peptides substituted with short N-glycan structures. 28433761 6 102 link N-linked 1181:1188 arg1 oligosaccharides 1190:1205 N-linked oligosaccharides 1181:1205 N-linked oligosaccharides 1181:1205 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 7 103 theme N-linked 1267:1274 arg1 chains 1276:1281 mainly short N-linked chains 1254:1281 mainly short N-linked chains 1254:1281 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 12 104 from site 2004:2007 arg1 protein 2038:2044 this protein 2033:2044 this protein 2033:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 5 105 theme sequence 783:790 arg1 SSASSASSA 792:800 the peptide sequence SSASSASSA 771:800 two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure 742:1039 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 13 106 theme chains 2156:2161 arg1 this 2110:2113 this 2110:2113 this 2110:2113 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 13 106 theme chains 2156:2161 arg1 description 2128:2138 the first description 2118:2138 the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp 2118:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 12 107 theme site 2004:2007 arg1 characterization 1975:1990 characterization 1975:1990 characterization of type and site of glycosidic chains in this protein 1975:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 3 108 theme exchange 429:436 arg1 chromatography 462:475 anion exchange high performance liquid chromatography 423:475 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 0 109 theme glycoproteomic 2:15 arg1 approach 17:24 A glycoproteomic approach 0:24 A glycoproteomic approach 0:24 A glycoproteomic approach reveals that the S-layer glycoprotein of Lactobacillus kefiri CIDCA 83111 is O- and N-glycosylated. 28433761 13 110 theme S-layer 2166:2172 arg1 glycoprotein 2174:2185 S-layer glycoprotein 2166:2185 S-layer glycoprotein from Lactobacillus spp 2166:2208 Furthermore, this is the first description of N-glycosidic chains in S-layer glycoprotein from Lactobacillus spp. 28433761 12 111 theme glycosidic 2012:2021 arg1 chains 2023:2028 glycosidic chains 2012:2028 glycosidic chains in this protein 2012:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 5 112 theme peptide 995:1001 arg1 471-476 1003:1009 peptide 471-476 995:1009 peptide 471-476 995:1009 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 6 113 theme acid 1121:1124 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 7 114 theme exchange 1214:1221 arg1 chromatography 1223:1236 Anion exchange chromatography 1208:1236 Anion exchange chromatography analysis 1208:1245 Anion exchange chromatography analysis showed mainly short N-linked chains. 28433761 6 115 theme S-layer 1107:1113 arg1 sequence 1126:1133 the S-layer amino acid sequence 1103:1133 the S-layer amino acid sequence 1103:1133 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 3 116 theme chromatography 462:475 arg1 analysis 477:484 anion exchange high performance liquid chromatography analysis 423:484 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 5 117 theme signature 826:834 arg1 motif 850:854 a signature glycosylation motif 824:854 a signature glycosylation motif 824:854 Further nanoHPLC-ESI analysis of the glycopeptides showed two O-glycosylated peptides: the peptide sequence SSASSASSA already identified as a signature glycosylation motif in L. buchneri, substituted on average with eight glucose residues and decorated with galacturonic acid and another O-glycosylated site on peptide 471-476, with a Glc5-8GalA2 structure. 28433761 12 118 theme type 1995:1998 arg1 characterization 1975:1990 characterization 1975:1990 characterization of type and site of glycosidic chains in this protein 1975:2044 Therefore, characterization of type and site of glycosidic chains in this protein may help to understand these important properties. 28433761 4 119 theme MALDI-TOF 542:550 arg1 analysis 570:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 3 120 theme performance 443:453 arg1 chromatography 462:475 anion exchange high performance liquid chromatography 423:475 anion exchange high performance liquid chromatography analysis 423:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 9 121 from description 1464:1474 arg1 glycoproteins 1517:1529 S-layer glycoproteins 1509:1529 S-layer glycoproteins from Lactobacillus species 1509:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 9 121 from description 1464:1474 arg1 species 1550:1556 Lactobacillus species 1536:1556 Lactobacillus species 1536:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 3 122 theme reductive 377:385 arg1 useful 490:495 useful 490:495 useful 490:495 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 3 122 theme reductive 377:385 arg1 treatment 401:409 A reductive β-elimination treatment 375:409 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis 375:484 A reductive β-elimination treatment followed by anion exchange high performance liquid chromatography analysis was useful to characterize the O-glycosidic structures. 28433761 4 123 theme spectrometry 557:568 arg1 analysis 570:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis 542:577 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 28433761 11 124 theme cells 1835:1839 arg1 interaction 1810:1820 the interaction 1806:1820 the interaction of bacterial cells with yeasts present in kefir grains 1806:1875 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 9 125 from species 1550:1556 arg1 description 1464:1474 the first description 1454:1474 the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species 1454:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 9 125 from species 1550:1556 arg1 glycoproteins 1517:1529 S-layer glycoproteins 1509:1529 S-layer glycoproteins from Lactobacillus species 1509:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 9 125 from species 1550:1556 arg1 this 1446:1449 this 1446:1449 this 1446:1449 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 2 126 gly glycosylation 301:313 arg1 protein 330:336 the S-layer protein 318:336 the S-layer protein of Lactobacillus kefiri CIDCA 83111 318:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 2 126 gly glycosylation 301:313 arg1 detail 290:295 detail 290:295 detail 290:295 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 11 127 theme pathogens 1953:1961 arg1 effects 1924:1930 the adverse effects 1912:1930 the adverse effects of different enteric pathogens 1912:1961 It is known that S-layer proteins from kefir-isolated L. kefiri strains are involved in the interaction of bacterial cells with yeasts present in kefir grains and are also capable to antagonize the adverse effects of different enteric pathogens. 28433761 9 128 theme first 1458:1462 arg1 description 1464:1474 the first description 1454:1474 the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species 1454:1556 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 9 128 theme first 1458:1462 arg1 this 1446:1449 this 1446:1449 this 1446:1449 To our knowledge, this is the first description of the structure of N-glycans in S-layer glycoproteins from Lactobacillus species. 28433761 6 129 theme PNGase 1151:1156 arg1 digestion 1160:1168 a PNGase F digestion 1149:1168 a PNGase F digestion 1149:1168 As ten characteristic sequons (Asn-X-Ser/Thr) are present in the S-layer amino acid sequence, we performed a PNGase F digestion to release N-linked oligosaccharides. 28433761 2 130 theme 83111 368:372 arg1 protein 330:336 the S-layer protein 318:336 the S-layer protein of Lactobacillus kefiri CIDCA 83111 318:372 The aim of this work was to characterize in detail the glycosylation of the S-layer protein of Lactobacillus kefiri CIDCA 83111. 28433761 4 131 theme glucose 642:648 arg1 units 650:654 5 to 8 glucose units 635:654 5 to 8 glucose units carrying galacturonic acid 635:681 MALDI-TOF mass spectrometry analysis confirmed the presence of oligosaccharides bearing from 5 to 8 glucose units carrying galacturonic acid. 25452313 8 0 gly sialylated 996:1005 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 5 1 theme Novel 526:530 arg1 biomarkers 532:541 Novel biomarkers 526:541 Novel biomarkers 526:541 Novel biomarkers are essential to identify patients for targeted and individualized therapy. 25452313 8 2 from common 1076:1081 arg1 adenomas 1105:1112 adenomas 1105:1112 adenomas 1105:1112 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 8 2 from common 1076:1081 arg1 carcinomas 1086:1095 carcinomas 1086:1095 carcinomas 1086:1095 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 10 3 theme tissue 1322:1327 arg1 expression 1329:1338 their tissue expression 1316:1338 their tissue expression in 220 colorectal cancer patients 1316:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 9 4 dep stage 1149:1153 arg1 carcinomas 1174:1183 carcinomas 1174:1183 carcinomas 1174:1183 We also found differences between stage I-II and stage III carcinomas. 25452313 1 5 gly glycoproteins 142:154 arg1 glycoproteins 142:154 glycoproteins 142:154 glycoproteins 142:154 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 8 6 theme N-glycan 1046:1053 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 12 7 theme related 1634:1640 arg1 glycans 1642:1648 several carcinoma related glycans 1616:1648 several carcinoma related glycans 1616:1648 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 12 8 theme biomarker 1773:1781 arg1 expression 1783:1792 biomarker expression 1773:1792 biomarker expression in patient samples 1773:1811 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 10 9 theme Lewis 1271:1275 arg1 a 1277:1277 sialyl Lewis a 1264:1277 sialyl Lewis a 1264:1277 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 8 10 theme pauci-mannose 1008:1020 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 5 11 theme individualized 595:608 arg1 therapy 610:616 targeted and individualized therapy 582:616 targeted and individualized therapy 582:616 Novel biomarkers are essential to identify patients for targeted and individualized therapy. 25452313 12 12 from expression 1783:1792 arg1 samples 1805:1811 patient samples 1797:1811 patient samples 1797:1811 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 10 13 theme sialyl 1264:1269 arg1 a 1277:1277 sialyl Lewis a 1264:1277 sialyl Lewis a 1264:1277 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 11 14 theme advanced 1474:1481 arg1 cancer 1494:1499 advanced colorectal cancer 1474:1499 advanced colorectal cancer 1474:1499 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 2 15 theme essential 259:267 arg1 events 252:257 events 252:257 events essential for cell viability and function 252:299 Most glycans are localized to cell surfaces and participate in events essential for cell viability and function. 25452313 6 16 theme stages 715:720 arg1 profiles 644:651 the N-glycan profiles 631:651 the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages 631:720 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 10 17 theme cancer 1358:1363 arg1 patients 1365:1372 220 colorectal cancer patients 1343:1372 220 colorectal cancer patients 1343:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 10 18 from analysis 1304:1311 arg1 patients 1365:1372 220 colorectal cancer patients 1343:1372 220 colorectal cancer patients 1343:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 6 19 theme different 705:713 arg1 stages 715:720 different stages 705:720 different stages 705:720 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 1 20 theme glycoproteins 142:154 arg1 glycans 107:113 glycans 107:113 glycans 107:113 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 1 20 theme glycoproteins 142:154 arg1 units 133:137 the carbohydrate units 116:137 the carbohydrate units of glycoproteins, glycolipids, and proteoglycans 116:186 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 0 21 theme separate 34:41 arg1 adenomas 50:57 separate rectal adenomas 34:57 separate rectal adenomas from carcinomas 34:73 N-glycomic profiling as a tool to separate rectal adenomas from carcinomas. 25452313 4 22 theme cancers 488:494 arg1 one 451:453 one 451:453 one 451:453 Colorectal cancer is one of the world's three most common cancers, and its incidence is rising. 25452313 4 22 theme cancers 488:494 arg1 cancers 488:494 the world's three most common cancers 458:494 the world's three most common cancers 458:494 Colorectal cancer is one of the world's three most common cancers, and its incidence is rising. 25452313 11 23 theme colorectal 1378:1387 arg1 cancer 1389:1394 colorectal cancer 1378:1394 colorectal cancer 1378:1394 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 11 24 theme colorectal 1483:1492 arg1 cancer 1494:1499 advanced colorectal cancer 1474:1499 advanced colorectal cancer 1474:1499 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 11 25 theme poor 1502:1505 arg1 prognosis 1507:1515 poor prognosis 1502:1515 poor prognosis 1502:1515 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 11 26 theme poor 1397:1400 arg1 prognosis 1402:1410 poor prognosis 1397:1410 poor prognosis 1397:1410 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 4 27 theme common 481:486 arg1 cancers 488:494 the world's three most common cancers 458:494 the world's three most common cancers 458:494 Colorectal cancer is one of the world's three most common cancers, and its incidence is rising. 25452313 6 28 theme carcinomas 691:700 arg1 profiles 644:651 the N-glycan profiles 631:651 the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages 631:720 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 1 29 theme glycolipids 157:167 arg1 glycans 107:113 glycans 107:113 glycans 107:113 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 1 29 theme glycolipids 157:167 arg1 units 133:137 the carbohydrate units 116:137 the carbohydrate units of glycoproteins, glycolipids, and proteoglycans 116:186 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 12 30 theme patient 1797:1803 arg1 samples 1805:1811 patient samples 1797:1811 patient samples 1797:1811 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 1 31 theme human 80:84 arg1 cells 86:90 All human cells 76:90 All human cells 76:90 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 6 32 theme rectal 684:689 arg1 carcinomas 691:700 18 rectal carcinomas 681:700 18 rectal carcinomas 681:700 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 3 33 theme carcinoma-related 361:377 arg1 structures 386:395 therefore carcinoma-related glycan structures 351:395 therefore carcinoma-related glycan structures 351:395 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 3 33 theme carcinoma-related 361:377 arg1 biomarkers 418:427 potential cancer biomarkers 401:427 potential cancer biomarkers 401:427 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 8 34 from adenomas 1105:1112 arg1 common 1076:1081 common 1076:1081 common 1076:1081 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 5 35 theme targeted 582:589 arg1 therapy 610:616 targeted and individualized therapy 582:616 targeted and individualized therapy 582:616 Novel biomarkers are essential to identify patients for targeted and individualized therapy. 25452313 10 36 from expression 1329:1338 arg1 patients 1365:1372 220 colorectal cancer patients 1343:1372 220 colorectal cancer patients 1343:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 11 37 theme elevated 1533:1540 arg1 expression 1542:1551 elevated expression 1533:1551 elevated expression of pauci-mannose 1533:1568 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 3 38 theme glycan 379:384 arg1 structures 386:395 therefore carcinoma-related glycan structures 351:395 therefore carcinoma-related glycan structures 351:395 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 3 38 theme glycan 379:384 arg1 biomarkers 418:427 potential cancer biomarkers 401:427 potential cancer biomarkers 401:427 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 2 39 theme cell 273:276 arg1 viability 278:286 cell viability 273:286 cell viability 273:286 Most glycans are localized to cell surfaces and participate in events essential for cell viability and function. 25452313 1 40 theme proteoglycans 174:186 arg1 glycans 107:113 glycans 107:113 glycans 107:113 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 1 40 theme proteoglycans 174:186 arg1 units 133:137 the carbohydrate units 116:137 the carbohydrate units of glycoproteins, glycolipids, and proteoglycans 116:186 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 11 41 theme elevated 1428:1435 arg1 expression 1437:1446 elevated expression 1428:1446 elevated expression of sialyl Lewis a 1428:1464 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 10 42 theme colorectal 1347:1356 arg1 patients 1365:1372 220 colorectal cancer patients 1343:1372 220 colorectal cancer patients 1343:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 10 43 theme expression 1329:1338 arg1 analysis 1304:1311 immunohistochemical analysis 1284:1311 immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients 1284:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 0 44 from carcinomas 64:73 arg1 adenomas 50:57 separate rectal adenomas 34:57 separate rectal adenomas from carcinomas 34:73 N-glycomic profiling as a tool to separate rectal adenomas from carcinomas. 25452313 6 45 theme adenomas 668:675 arg1 profiles 644:651 the N-glycan profiles 631:651 the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages 631:720 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 11 46 theme pauci-mannose 1556:1568 arg1 expression 1542:1551 elevated expression 1533:1551 elevated expression of pauci-mannose 1533:1568 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 12 47 theme mass 1589:1592 arg1 spectrometry 1594:1605 mass spectrometry 1589:1605 mass spectrometry 1589:1605 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 3 48 theme potential 401:409 arg1 structures 386:395 therefore carcinoma-related glycan structures 351:395 therefore carcinoma-related glycan structures 351:395 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 3 48 theme potential 401:409 arg1 biomarkers 418:427 potential cancer biomarkers 401:427 potential cancer biomarkers 401:427 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 11 49 theme a 1464:1464 arg1 expression 1437:1446 elevated expression 1428:1446 elevated expression of sialyl Lewis a 1428:1464 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 8 50 theme monoantennary 981:993 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 3 51 theme cancer 411:416 arg1 structures 386:395 therefore carcinoma-related glycan structures 351:395 therefore carcinoma-related glycan structures 351:395 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 3 51 theme cancer 411:416 arg1 biomarkers 418:427 potential cancer biomarkers 401:427 potential cancer biomarkers 401:427 Glycosylation evolves during carcinogenesis, and therefore carcinoma-related glycan structures are potential cancer biomarkers. 25452313 7 52 theme Paraffin-embedded 803:819 arg1 samples 827:833 Paraffin-embedded tumor samples 803:833 Paraffin-embedded tumor samples 803:833 Paraffin-embedded tumor samples were deparaffinized, and glycans were enzymatically released and purified. 25452313 2 53 theme Most 189:192 arg1 glycans 194:200 Most glycans 189:200 Most glycans 189:200 Most glycans are localized to cell surfaces and participate in events essential for cell viability and function. 25452313 8 54 from differences 919:929 arg1 glycosylation 934:946 glycosylation 934:946 glycosylation 934:946 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 6 55 theme mass 784:787 arg1 spectrometry 789:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 6 56 theme rectal 661:666 arg1 adenomas 668:675 five rectal adenomas 656:675 five rectal adenomas 656:675 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 7 57 theme tumor 821:825 arg1 samples 827:833 Paraffin-embedded tumor samples 803:833 Paraffin-embedded tumor samples 803:833 Paraffin-embedded tumor samples were deparaffinized, and glycans were enzymatically released and purified. 25452313 4 58 theme Colorectal 430:439 arg1 cancer 441:446 Colorectal cancer 430:446 Colorectal cancer 430:446 Colorectal cancer is one of the world's three most common cancers, and its incidence is rising. 25452313 8 59 theme high-mannose 1033:1044 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 11 60 theme sialyl 1451:1456 arg1 a 1464:1464 sialyl Lewis a 1451:1464 sialyl Lewis a 1451:1464 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 8 61 theme small 1027:1031 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 12 62 theme carcinoma 1624:1632 arg1 glycans 1642:1648 several carcinoma related glycans 1616:1648 several carcinoma related glycans 1616:1648 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 8 63 theme sialylated 996:1005 arg1 structures 1055:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures 981:1064 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 10 64 from patients 1365:1372 arg1 analysis 1304:1311 immunohistochemical analysis 1284:1311 immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients 1284:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 6 65 theme desorption-ionization 747:767 arg1 spectrometry 789:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 12 66 theme several 1616:1622 arg1 glycans 1642:1648 several carcinoma related glycans 1616:1648 several carcinoma related glycans 1616:1648 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 11 67 theme Lewis 1458:1462 arg1 a 1464:1464 sialyl Lewis a 1451:1464 sialyl Lewis a 1451:1464 In colorectal cancer, poor prognosis correlated with elevated expression of sialyl Lewis a, and in advanced colorectal cancer, poor prognosis correlated with elevated expression of pauci-mannose. 25452313 6 68 theme laser 741:745 arg1 spectrometry 789:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 8 69 from carcinomas 1086:1095 arg1 common 1076:1081 common 1076:1081 common 1076:1081 We found differences in glycosylation between adenomas and carcinomas: monoantennary, sialylated, pauci-mannose, and small high-mannose N-glycan structures were more common in carcinomas than in adenomas. 25452313 10 70 theme immunohistochemical 1284:1302 arg1 analysis 1304:1311 immunohistochemical analysis 1284:1311 immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients 1284:1372 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 10 71 dep structures 1234:1243 arg1 pauci-mannose 1246:1258 pauci-mannose 1246:1258 pauci-mannose 1246:1258 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 10 71 dep structures 1234:1243 arg1 a 1277:1277 sialyl Lewis a 1264:1277 sialyl Lewis a 1264:1277 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25452313 0 72 theme rectal 43:48 arg1 adenomas 50:57 separate rectal adenomas 34:57 separate rectal adenomas from carcinomas 34:73 N-glycomic profiling as a tool to separate rectal adenomas from carcinomas. 25452313 12 73 theme applicable 1746:1755 arg1 method 1757:1762 a readily applicable method 1736:1762 a readily applicable method to study biomarker expression in patient samples 1736:1811 In conclusion, by mass spectrometry we found several carcinoma related glycans, and we demonstrate a method of transforming these results into immunohistochemistry, a readily applicable method to study biomarker expression in patient samples. 25452313 2 74 theme cell 219:222 arg1 surfaces 224:231 cell surfaces 219:231 cell surfaces 219:231 Most glycans are localized to cell surfaces and participate in events essential for cell viability and function. 25452313 6 75 theme matrix-assisted 725:739 arg1 spectrometry 789:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 matrix-assisted laser desorption-ionization time-of-flight mass spectrometry 725:800 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 1 76 theme carbohydrate 120:131 arg1 glycans 107:113 glycans 107:113 glycans 107:113 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 1 76 theme carbohydrate 120:131 arg1 units 133:137 the carbohydrate units 116:137 the carbohydrate units of glycoproteins, glycolipids, and proteoglycans 116:186 All human cells are covered by glycans, the carbohydrate units of glycoproteins, glycolipids, and proteoglycans. 25452313 6 77 theme N-glycan 635:642 arg1 profiles 644:651 the N-glycan profiles 631:651 the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages 631:720 We compared the N-glycan profiles of five rectal adenomas and 18 rectal carcinomas of different stages by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. 25452313 10 78 theme glycan 1227:1232 arg1 structures 1234:1243 two glycan structures 1223:1243 two glycan structures: pauci-mannose and sialyl Lewis a 1223:1277 Based on these findings, we selected two glycan structures: pauci-mannose and sialyl Lewis a, for immunohistochemical analysis of their tissue expression in 220 colorectal cancer patients. 25348702 7 0 theme deeper 1113:1118 arg1 insight 1120:1126 deeper insight 1113:1126 deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation 1113:1219 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 1 theme N-glycan 1152:1159 arg1 fucosylation 1191:1202 fucosylation 1191:1202 fucosylation 1191:1202 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 1 theme N-glycan 1152:1159 arg1 features 1161:1168 cell-type-specific N-glycan features 1133:1168 cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation 1133:1219 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 1 theme N-glycan 1152:1159 arg1 antennarity 1178:1188 antennarity 1178:1188 antennarity 1178:1188 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 1 theme N-glycan 1152:1159 arg1 sialylation 1209:1219 sialylation 1209:1219 sialylation 1209:1219 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 8 2 from useful 1298:1303 arg1 fields 1312:1317 the fields 1308:1317 the fields of glycobiotechnology and biomarker discovery 1308:1363 Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery. 25348702 0 3 theme mass 71:74 arg1 spectrometry 76:87 mass spectrometry 71:87 mass spectrometry 71:87 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 0 4 from analysis 6:13 arg1 cells 59:63 living cells 52:63 living cells 52:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 2 5 theme cells 331:335 arg1 glycome 313:319 the glycome 309:319 the glycome of living cells 309:335 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 6 6 from 42 1043:1044 arg1 CHO-K1 1049:1054 CHO-K1 1049:1054 CHO-K1 1049:1054 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 3 7 theme common 357:362 arg1 procedure 364:372 a common procedure 355:372 a common procedure 355:372 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 8 theme relative 928:935 arg1 amount 937:942 their relative amount 922:942 their relative amount from 14 to 85% 922:957 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 3 9 theme intracellular 520:532 arg1 proteins 534:541 intracellular proteins 520:541 intracellular proteins derived from the ER 520:561 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 10 theme N-glycans 898:906 arg1 quantification 867:880 quantification 867:880 quantification 867:880 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 5 10 theme N-glycans 898:906 arg1 detection 853:861 detection 853:861 detection 853:861 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 4 11 from cells 706:710 arg1 N-glycosylation 678:692 profile surface N-glycosylation 662:692 profile surface N-glycosylation from living cells 662:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 4 12 theme living 699:704 arg1 cells 706:710 living cells 699:710 living cells 699:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 3 13 attach derived 543:549 arg1 ER 560:561 the ER 556:561 the ER 556:561 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 3 13 attach derived 543:549 arg2 proteins 534:541 intracellular proteins 520:541 intracellular proteins derived from the ER 520:561 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 14 theme complex-type 885:896 arg1 N-glycans 898:906 complex-type N-glycans 885:906 complex-type N-glycans 885:906 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 4 15 theme sensitive 632:640 arg1 method 652:657 a reproducible, sensitive, and fast method 616:657 a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells 616:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 4 16 theme fast 647:650 arg1 method 652:657 a reproducible, sensitive, and fast method 616:657 a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells 616:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 6 17 theme G2 1071:1072 arg1 cells 1074:1078 Hep G2 cells 1067:1078 Hep G2 cells 1067:1078 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 7 18 theme additional 1085:1094 arg1 signals 1096:1102 The additional signals 1081:1102 The additional signals 1081:1102 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 3 19 theme cell 383:386 arg1 lysis 388:392 cell lysis 383:392 cell lysis 383:392 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 2 20 theme paramount 254:262 arg1 importance 264:273 paramount importance 254:273 paramount importance 254:273 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 3 21 theme membrane 412:419 arg1 proteins 428:435 membrane (glyco)proteins 412:435 membrane (glyco)proteins 412:435 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 22 theme cell 753:756 arg1 surfaces 758:765 cell surfaces 753:765 cell surfaces 753:765 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 5 23 from surfaces 758:765 arg1 glycopeptides 734:746 glycopeptides 734:746 glycopeptides from cell surfaces 734:765 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 4 24 theme surface 670:676 arg1 N-glycosylation 678:692 profile surface N-glycosylation 662:692 profile surface N-glycosylation from living cells 662:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 3 25 theme high-mannose 470:481 arg1 N-glycans 483:491 high-mannose N-glycans 470:491 high-mannose N-glycans 470:491 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 26 theme cells 824:828 arg1 digestion 783:791 tryptic digestion 775:791 tryptic digestion of freshly harvested and vital cells 775:828 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 0 27 theme Rapid 0:4 arg1 analysis 6:13 Rapid analysis 0:13 Rapid analysis of cell surface N-glycosylation from living cells 0:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 1 28 theme Cell 90:93 arg1 surfaces 95:102 Cell surfaces 90:102 Cell surfaces 90:102 Cell surfaces are covered with a dense carbohydrate layer referred to as the glycocalyx. 25348702 0 29 theme cell 18:21 arg1 N-glycosylation 31:45 cell surface N-glycosylation 18:45 cell surface N-glycosylation from living cells 18:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 8 30 theme biomarker 1345:1353 arg1 discovery 1355:1363 biomarker discovery 1345:1363 biomarker discovery 1345:1363 Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery. 25348702 8 31 from fields 1312:1317 arg1 useful 1298:1303 useful 1298:1303 useful 1298:1303 Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery. 25348702 6 32 theme additional 994:1003 arg1 structures 1005:1014 25 additional structures 991:1014 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 991:1062 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 4 33 theme profile 662:668 arg1 N-glycosylation 678:692 profile surface N-glycosylation 662:692 profile surface N-glycosylation from living cells 662:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 2 34 theme robust 283:288 arg1 methods 290:296 robust methods 283:296 robust methods 283:296 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 3 35 theme proteins 428:435 arg1 extraction 398:407 extraction 398:407 extraction 398:407 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 3 35 theme proteins 428:435 arg1 lysis 388:392 cell lysis 383:392 cell lysis 383:392 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 0 36 theme N-glycosylation 31:45 arg1 analysis 6:13 Rapid analysis 0:13 Rapid analysis of cell surface N-glycosylation from living cells 0:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 2 37 contain have 278:281 arg1 it 245:246 it 245:246 it 245:246 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 2 37 contain have 278:281 arg2 methods 290:296 robust methods 283:296 robust methods 283:296 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 8 38 theme glycobiotechnology 1322:1339 arg1 fields 1312:1317 the fields 1308:1317 the fields of glycobiotechnology and biomarker discovery 1308:1363 Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery. 25348702 0 39 theme surface 23:29 arg1 N-glycosylation 31:45 cell surface N-glycosylation 18:45 cell surface N-glycosylation from living cells 18:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 5 40 dep 85 955:956 arg1 to 952:953 to 952:953 to 952:953 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 2 41 theme glycan 226:231 arg1 signatures 233:242 different glycan signatures 216:242 different glycan signatures 216:242 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 5 42 theme harvested 804:812 arg1 cells 824:828 freshly harvested and vital cells 796:828 freshly harvested and vital cells 796:828 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 3 43 theme glyco 422:426 arg1 proteins 428:435 membrane (glyco)proteins 412:435 membrane (glyco)proteins 412:435 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 0 44 theme living 52:57 arg1 cells 59:63 living cells 52:63 living cells 52:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 0 45 from cells 59:63 arg1 N-glycosylation 31:45 cell surface N-glycosylation 18:45 cell surface N-glycosylation from living cells 18:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 0 45 from cells 59:63 arg1 analysis 6:13 Rapid analysis 0:13 Rapid analysis of cell surface N-glycosylation from living cells 0:63 Rapid analysis of cell surface N-glycosylation from living cells using mass spectrometry. 25348702 2 46 theme different 216:224 arg1 signatures 233:242 different glycan signatures 216:242 different glycan signatures 216:242 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 5 47 dep detection 853:861 arg1 the 849:851 the 849:851 the 849:851 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 2 48 theme living 324:329 arg1 cells 331:335 living cells 324:335 living cells 324:335 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 3 49 theme major 450:454 arg1 proportion 456:465 a major proportion 448:465 a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER 448:561 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 25348702 5 50 from % 957:957 arg1 amount 937:942 their relative amount 922:942 their relative amount from 14 to 85% 922:957 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 2 51 theme cell 197:200 arg1 types 202:206 different cell types 187:206 different cell types 187:206 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 4 52 theme reproducible 618:629 arg1 method 652:657 a reproducible, sensitive, and fast method 616:657 a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells 616:710 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 4 53 theme HEK 570:572 arg1 system 595:600 a model system 587:600 a model system 587:600 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 4 53 theme HEK 570:572 arg1 cells 578:582 HEK 293 cells 570:582 HEK 293 cells 570:582 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 6 54 theme Hep 1067:1069 arg1 cells 1074:1078 Hep G2 cells 1067:1078 Hep G2 cells 1067:1078 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 2 55 theme different 187:195 arg1 types 202:206 different cell types 187:206 different cell types 187:206 Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. 25348702 5 56 theme tryptic 775:781 arg1 digestion 783:791 tryptic digestion 775:791 tryptic digestion of freshly harvested and vital cells 775:828 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 6 57 from 48 1028:1029 arg1 AGE1.HN 1034:1040 AGE1.HN 1034:1040 AGE1.HN 1034:1040 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 1 58 theme dense 123:127 arg1 layer 142:146 a dense carbohydrate layer 121:146 a dense carbohydrate layer referred to as the glycocalyx 121:176 Cell surfaces are covered with a dense carbohydrate layer referred to as the glycocalyx. 25348702 8 59 theme discovery 1355:1363 arg1 fields 1312:1317 the fields 1308:1317 the fields of glycobiotechnology and biomarker discovery 1308:1363 Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery. 25348702 4 60 theme model 589:593 arg1 system 595:600 a model system 587:600 a model system 587:600 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 4 60 theme model 589:593 arg1 cells 578:582 HEK 293 cells 570:582 HEK 293 cells 570:582 Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. 25348702 1 61 theme carbohydrate 129:140 arg1 layer 142:146 a dense carbohydrate layer 121:146 a dense carbohydrate layer referred to as the glycocalyx 121:176 Cell surfaces are covered with a dense carbohydrate layer referred to as the glycocalyx. 25348702 5 62 theme vital 818:822 arg1 cells 824:828 freshly harvested and vital cells 796:828 freshly harvested and vital cells 796:828 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 7 63 theme cell-type-specific 1133:1150 arg1 fucosylation 1191:1202 fucosylation 1191:1202 fucosylation 1191:1202 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 63 theme cell-type-specific 1133:1150 arg1 features 1161:1168 cell-type-specific N-glycan features 1133:1168 cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation 1133:1219 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 63 theme cell-type-specific 1133:1150 arg1 antennarity 1178:1188 antennarity 1178:1188 antennarity 1178:1188 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 7 63 theme cell-type-specific 1133:1150 arg1 sialylation 1209:1219 sialylation 1209:1219 sialylation 1209:1219 The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. 25348702 5 64 gly glycopeptides 734:746 arg1 surfaces 758:765 cell surfaces 753:765 cell surfaces 753:765 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 5 64 gly glycopeptides 734:746 arg2 glycopeptides 734:746 glycopeptides 734:746 glycopeptides from cell surfaces 734:765 We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. 25348702 6 65 from structures 1005:1014 arg1 51 1061:1062 51 1061:1062 51 1061:1062 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 6 65 from structures 1005:1014 arg1 42 1043:1044 42 1043:1044 42 1043:1044 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 6 65 from structures 1005:1014 arg1 HEK 1019:1021 HEK 293 1019:1025 HEK 293 1019:1025 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 6 65 from structures 1005:1014 arg1 48 1028:1029 48 1028:1029 48 1028:1029 It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. 25348702 3 66 theme N-glycans 483:491 arg1 proportion 456:465 a major proportion 448:465 a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER 448:561 To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. 28921955 0 0 theme Spodoptera 95:104 arg1 Line 129:132 Spodoptera frugiperda Insect Cell Line 95:132 Spodoptera frugiperda Insect Cell Line 95:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 5 1 gly glycopeptide 910:921 arg2 glycopeptide 910:921 a given glycopeptide 902:921 a given glycopeptide 902:921 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 5 2 theme collision-induced 941:957 arg1 dissociation 959:970 collision-induced dissociation 941:970 collision-induced dissociation MS/MS fragmentation 941:990 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 3 theme glycan 728:733 arg1 occupancy 735:743 glycan occupancy 728:743 glycan occupancy 728:743 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 1 4 theme related 185:191 arg1 hemagglutinin 145:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 1 4 theme related 185:191 arg1 glycoprotein 172:183 a surface glycoprotein 162:183 a surface glycoprotein related to virus invasion and host immune system response 162:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 0 5 theme Insect 117:122 arg1 Line 129:132 Spodoptera frugiperda Insect Cell Line 95:132 Spodoptera frugiperda Insect Cell Line 95:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 0 6 from Characterization 0:15 arg1 Hemagglutinin 69:81 Influenza A Virus Hemagglutinin 51:81 Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 51:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 5 7 theme peptide 1001:1007 arg1 information 1018:1028 the peptide backbone information 997:1028 the peptide backbone information 997:1028 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 2 8 dep design 373:378 arg1 the 369:371 the 369:371 the 369:371 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 0 9 theme frugiperda 106:115 arg1 Line 129:132 Spodoptera frugiperda Insect Cell Line 95:132 Spodoptera frugiperda Insect Cell Line 95:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 7 10 theme glycosylation 1191:1203 arg1 sites 1205:1209 Six glycosylation sites 1187:1209 Six glycosylation sites fitting the sequon N-X-S/T 1187:1236 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 5 11 theme backbone 1009:1016 arg1 information 1018:1028 the peptide backbone information 997:1028 the peptide backbone information 997:1028 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 5 12 theme MS/MS 972:976 arg1 fragmentation 978:990 collision-induced dissociation MS/MS fragmentation 941:990 collision-induced dissociation MS/MS fragmentation 941:990 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 13 theme tandem 497:502 arg1 analysis 507:514 LC tandem MS analysis 494:514 LC tandem MS analysis 494:514 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 3 14 gly glycosylation 533:545 arg1 H1/A/California/04/2009 575:597 H1/A/California/04/2009 575:597 H1/A/California/04/2009 575:597 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 3 14 gly glycosylation 533:545 arg1 hemagglutinin 560:572 Influenza hemagglutinin 550:572 Influenza hemagglutinin (H1/A/California/04/2009) 550:598 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 5 15 theme dissociation 959:970 arg1 fragmentation 978:990 collision-induced dissociation MS/MS fragmentation 941:990 collision-induced dissociation MS/MS fragmentation 941:990 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 16 theme multienzyme 460:470 arg1 digestion 472:480 multienzyme digestion 460:480 multienzyme digestion followed by LC tandem MS analysis 460:514 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 0 17 theme Cell 124:127 arg1 Line 129:132 Spodoptera frugiperda Insect Cell Line 95:132 Spodoptera frugiperda Insect Cell Line 95:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 3 18 theme glycoproteomic 427:440 arg1 analysis 442:449 glycoproteomic analysis 427:449 glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis 427:514 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 6 19 theme sequence 1127:1134 arg1 coverage 1136:1143 100% sequence coverage 1122:1143 100% sequence coverage of the hemagglutinin sample 1122:1171 With this approach, 100% sequence coverage of the hemagglutinin sample was obtained. 28921955 1 20 theme virus 196:200 arg1 invasion 202:209 virus invasion 196:209 virus invasion 196:209 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 7 21 theme sequon 1223:1228 arg1 N-X-S/T 1230:1236 the sequon N-X-S/T 1219:1236 the sequon N-X-S/T 1219:1236 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 3 22 theme Influenza 550:558 arg1 H1/A/California/04/2009 575:597 H1/A/California/04/2009 575:597 H1/A/California/04/2009 575:597 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 3 22 theme Influenza 550:558 arg1 hemagglutinin 560:572 Influenza hemagglutinin 550:572 Influenza hemagglutinin (H1/A/California/04/2009) 550:598 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 6 23 theme % 1125:1125 arg1 coverage 1136:1143 100% sequence coverage 1122:1143 100% sequence coverage of the hemagglutinin sample 1122:1171 With this approach, 100% sequence coverage of the hemagglutinin sample was obtained. 28921955 3 24 theme following 610:618 arg1 steps 620:624 the following steps 606:624 the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy 606:743 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 7 25 gly glycosylation 1191:1203 arg2 sites 1205:1209 Six glycosylation sites 1187:1209 Six glycosylation sites fitting the sequon N-X-S/T 1187:1236 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 7 25 gly glycosylation 1191:1203 arg2 Six 1187:1189 Six 1187:1189 Six 1187:1189 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 7 26 gly heterogeneity 1282:1294 arg1 glycoforms 1321:1330 glycoforms 1321:1330 glycoforms 1321:1330 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 5 27 dep dissociation 1064:1075 arg1 fragmentation 1087:1099 -MS3 fragmentation 1082:1099 collision-induced dissociation (CID)-MS3 fragmentation 1046:1099 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 2 28 gly glycosylation 272:284 arg1 hemagglutinin 289:301 hemagglutinin 289:301 hemagglutinin 289:301 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 4 29 theme LC 855:856 arg1 analysis 868:875 online LC tandem MS analysis 848:875 online LC tandem MS analysis 848:875 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 30 theme online 848:853 arg1 analysis 868:875 online LC tandem MS analysis 848:875 online LC tandem MS analysis 848:875 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 1 31 theme host 215:218 arg1 response 234:241 host immune system response 215:241 host immune system response 215:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 0 32 theme Site-Specific 20:32 arg1 Glycosylation 34:46 Site-Specific Glycosylation 20:46 Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 20:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 3 33 theme hemagglutinin 560:572 arg1 glycosylation 533:545 the glycosylation 529:545 the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) 529:598 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 2 34 theme virus 337:341 arg1 evolution 343:351 virus evolution 337:351 virus evolution 337:351 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 3 35 dep steps 620:624 arg1 used 691:694 used 691:694 was used to determine the glycosites and glycan occupancy 687:743 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 1 36 theme immune 220:225 arg1 response 234:241 host immune system response 215:241 host immune system response 215:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 0 37 theme Influenza 51:59 arg1 Hemagglutinin 69:81 Influenza A Virus Hemagglutinin 51:81 Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 51:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 3 38 used used 691:694 arg2 treatment 635:643 PNGaseF treatment 627:643 PNGaseF treatment combined with trypsin or pepsin digestion 627:685 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 2 39 theme specific 263:270 arg1 glycosylation 272:284 site specific glycosylation 258:284 site specific glycosylation of hemagglutinin 258:301 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 1 40 theme system 227:232 arg1 response 234:241 host immune system response 215:241 host immune system response 215:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 5 41 theme glycan 882:887 arg1 structure 889:897 The glycan structure 878:897 The glycan structure of a given glycopeptide 878:921 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 42 theme pepsin 670:675 arg1 digestion 677:685 trypsin or pepsin digestion 659:685 trypsin or pepsin digestion 659:685 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 0 43 theme Glycosylation 34:46 arg1 Characterization 0:15 Characterization 0:15 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 0:133 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 0 44 theme Virus 63:67 arg1 Hemagglutinin 69:81 Influenza A Virus Hemagglutinin 51:81 Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 51:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 5 45 theme given 904:908 arg1 glycopeptide 910:921 a given glycopeptide 902:921 a given glycopeptide 902:921 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 46 used used 422:425 arg2 we 419:420 we 419:420 we 419:420 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 0 47 from Hemagglutinin 69:81 arg1 Characterization 0:15 Characterization 0:15 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 0:133 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 4 48 theme tandem 858:863 arg1 analysis 868:875 online LC tandem MS analysis 848:875 online LC tandem MS analysis 848:875 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 6 49 theme sample 1166:1171 arg1 coverage 1136:1143 100% sequence coverage 1122:1143 100% sequence coverage of the hemagglutinin sample 1122:1171 With this approach, 100% sequence coverage of the hemagglutinin sample was obtained. 28921955 7 50 theme glycoforms 1321:1330 arg1 ratios 1311:1316 the ratios 1307:1316 the glycan heterogeneity as well as the ratios of glycoforms 1271:1330 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 7 50 theme glycoforms 1321:1330 arg1 heterogeneity 1282:1294 the glycan heterogeneity 1271:1294 the glycan heterogeneity as well as the ratios of glycoforms 1271:1330 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 0 51 theme A 61:61 arg1 Hemagglutinin 69:81 Influenza A Virus Hemagglutinin 51:81 Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 51:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 4 52 theme MS 865:866 arg1 analysis 868:875 online LC tandem MS analysis 848:875 online LC tandem MS analysis 848:875 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 5 53 theme -MS3 1082:1085 arg1 fragmentation 1087:1099 -MS3 fragmentation 1082:1099 collision-induced dissociation (CID)-MS3 fragmentation 1046:1099 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 54 theme PNGaseF 627:633 arg1 treatment 635:643 PNGaseF treatment 627:643 PNGaseF treatment combined with trypsin or pepsin digestion 627:685 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 1 55 theme Influenza 135:143 arg1 hemagglutinin 145:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 1 55 theme Influenza 135:143 arg1 glycoprotein 172:183 a surface glycoprotein 162:183 a surface glycoprotein related to virus invasion and host immune system response 162:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 4 56 used used 793:796 arg2 AspN 770:773 AspN 770:773 AspN 770:773 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 56 used used 793:796 arg2 trypsin 761:767 trypsin 761:767 trypsin 761:767 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 56 used used 793:796 arg2 pepsin 780:785 pepsin 780:785 pepsin 780:785 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 56 used used 793:796 arg2 enzymes 752:758 Three enzymes 746:758 Three enzymes 746:758 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 57 theme suitable 821:828 arg1 glycopeptides 830:842 suitable glycopeptides 821:842 suitable glycopeptides 821:842 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 4 58 gly glycopeptides 830:842 arg2 glycopeptides 830:842 suitable glycopeptides 821:842 suitable glycopeptides 821:842 Three enzymes, trypsin, AspN, and pepsin, were used separately to generate suitable glycopeptides for online LC tandem MS analysis. 28921955 3 59 theme MS 504:505 arg1 analysis 507:514 LC tandem MS analysis 494:514 LC tandem MS analysis 494:514 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 6 60 theme hemagglutinin 1152:1164 arg1 sample 1166:1171 the hemagglutinin sample 1148:1171 the hemagglutinin sample 1148:1171 With this approach, 100% sequence coverage of the hemagglutinin sample was obtained. 28921955 1 61 gly glycoprotein 172:183 arg1 hemagglutinin 145:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 1 61 gly glycoprotein 172:183 arg1 glycoprotein 172:183 a surface glycoprotein 162:183 a surface glycoprotein related to virus invasion and host immune system response 162:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 2 62 theme hemagglutinin 289:301 arg1 glycosylation 272:284 site specific glycosylation 258:284 site specific glycosylation of hemagglutinin 258:301 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 2 63 theme vaccines 395:402 arg1 design 373:378 design 373:378 design 373:378 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 2 63 theme vaccines 395:402 arg1 quality 384:390 quality 384:390 quality 384:390 Understanding site specific glycosylation of hemagglutinin will increase our knowledge about virus evolution and can improve the design and quality of vaccines. 28921955 3 64 gly glycosites 713:722 arg2 glycosites 713:722 the glycosites 709:722 the glycosites 709:722 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 7 65 theme glycan 1275:1280 arg1 heterogeneity 1282:1294 the glycan heterogeneity 1271:1294 the glycan heterogeneity as well as the ratios of glycoforms 1271:1330 Six glycosylation sites fitting the sequon N-X-S/T were successfully confirmed, and the glycan heterogeneity as well as the ratios of glycoforms were determined at each site. 28921955 5 66 theme collision-induced 1046:1062 arg1 CID 1078:1080 CID 1078:1080 CID 1078:1080 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 5 66 theme collision-induced 1046:1062 arg1 dissociation 1064:1075 collision-induced dissociation 1046:1075 collision-induced dissociation (CID)-MS3 fragmentation 1046:1099 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 3 67 theme LC 494:495 arg1 analysis 507:514 LC tandem MS analysis 494:514 LC tandem MS analysis 494:514 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 5 68 theme glycopeptide 910:921 arg1 structure 889:897 The glycan structure 878:897 The glycan structure of a given glycopeptide 878:921 The glycan structure of a given glycopeptide was determined by collision-induced dissociation MS/MS fragmentation, and the peptide backbone information was provided by collision-induced dissociation (CID)-MS3 fragmentation. 28921955 6 69 theme 100 1122:1124 arg1 % 1125:1125 % 1125:1125 % 1125:1125 With this approach, 100% sequence coverage of the hemagglutinin sample was obtained. 28921955 0 70 from Glycosylation 34:46 arg1 Hemagglutinin 69:81 Influenza A Virus Hemagglutinin 51:81 Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line 51:132 Characterization of Site-Specific Glycosylation in Influenza A Virus Hemagglutinin Produced by Spodoptera frugiperda Insect Cell Line. 28921955 3 71 theme trypsin 659:665 arg1 digestion 677:685 trypsin or pepsin digestion 659:685 trypsin or pepsin digestion 659:685 In our study, we used glycoproteomic analysis based on multienzyme digestion followed by LC tandem MS analysis to determine the glycosylation of Influenza hemagglutinin (H1/A/California/04/2009) using the following steps: PNGaseF treatment combined with trypsin or pepsin digestion was used to determine the glycosites and glycan occupancy. 28921955 1 72 theme surface 164:170 arg1 hemagglutinin 145:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin 135:157 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 28921955 1 72 theme surface 164:170 arg1 glycoprotein 172:183 a surface glycoprotein 162:183 a surface glycoprotein related to virus invasion and host immune system response 162:241 Influenza hemagglutinin is a surface glycoprotein related to virus invasion and host immune system response. 26132640 6 0 theme total 957:961 arg1 O-glycans 963:971 total O-glycans 957:971 total O-glycans 957:971 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 1 1 theme proteins 193:200 arg1 glycosylation 111:123 O-Linked glycosylation 102:123 O-Linked glycosylation of serine/threonine residues 102:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 1 theme proteins 193:200 arg1 modification 177:188 a posttranslational modification 157:188 a posttranslational modification of proteins 157:200 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 4 2 theme novel 621:625 arg1 procedure 639:647 a novel O-glycomics procedure 619:647 a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP) 619:713 Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP). 26132640 0 3 theme Analogues 91:99 arg1 Presence 68:75 the Presence 64:75 the Presence of Pyrazolone Analogues 64:99 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues. 26132640 0 4 from O-Glycomics 13:23 arg1 Presence 68:75 the Presence 64:75 the Presence of Pyrazolone Analogues 64:99 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues. 26132640 6 5 gly glycoproteins 984:996 arg1 glycoproteins 984:996 model glycoproteins 978:996 model glycoproteins (PSM) 978:1002 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 5 gly glycoproteins 984:996 arg1 PSM 999:1001 PSM 999:1001 PSM 999:1001 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 6 theme MW-assisted 1109:1119 arg1 procedure 1125:1133 this MW-assisted BEP procedure 1104:1133 this MW-assisted BEP procedure 1104:1133 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 1 7 gly glycosylation 111:123 arg1 residues 145:152 serine/threonine residues 128:152 serine/threonine residues 128:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 7 gly glycosylation 111:123 arg1 protein 223:229 protein recognition 223:241 protein recognition 223:241 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 7 gly glycosylation 111:123 arg1 lipid 247:251 lipid functions 247:261 lipid functions 247:261 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 6 8 theme MW-assisted 893:903 arg1 reaction 909:916 the MW-assisted BEP reaction 889:916 the MW-assisted BEP reaction 889:916 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 2 9 theme structured 334:343 arg1 glycans 354:360 differently structured O-linked glycans 322:360 differently structured O-linked glycans (O-glycans) 322:372 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 2 9 theme structured 334:343 arg1 O-glycans 363:371 O-glycans 363:371 O-glycans 363:371 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 2 10 theme O-linked 345:352 arg1 glycans 354:360 differently structured O-linked glycans 322:360 differently structured O-linked glycans (O-glycans) 322:372 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 2 10 theme O-linked 345:352 arg1 O-glycans 363:371 O-glycans 363:371 O-glycans 363:371 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 6 11 dep h. 1051:1052 arg1 to 1046:1047 to 1046:1047 to 1046:1047 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 12 theme BEP 1121:1123 arg1 procedure 1125:1133 this MW-assisted BEP procedure 1104:1133 this MW-assisted BEP procedure 1104:1133 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 4 13 theme O-glycomics 627:637 arg1 procedure 639:647 a novel O-glycomics procedure 619:647 a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP) 619:713 Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP). 26132640 6 14 theme various 1166:1172 arg1 samples 1185:1191 various biological samples 1166:1191 various biological samples 1166:1191 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 3 15 theme unwanted 572:579 arg1 reactions 586:594 unwanted side reactions 572:594 unwanted side reactions 572:594 Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. 26132640 2 16 theme such 448:451 arg1 groups 453:458 such groups 448:458 such groups 448:458 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 6 17 theme reaction 1012:1019 arg1 time 1021:1024 the reaction time 1008:1024 the reaction time 1008:1024 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 3 18 theme digestion 481:489 arg1 approaches 491:500 chemical digestion approaches 472:500 chemical digestion approaches 472:500 Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. 26132640 3 19 theme side 581:584 arg1 reactions 586:594 unwanted side reactions 572:594 unwanted side reactions 572:594 Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. 26132640 5 20 theme -assisted 766:774 arg1 procedure 780:788 a microwave (MW)-assisted BEP procedure 750:788 a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis 750:835 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 1 21 theme O-Linked 102:109 arg1 glycosylation 111:123 O-Linked glycosylation 102:123 O-Linked glycosylation of serine/threonine residues 102:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 21 theme O-Linked 102:109 arg1 modification 177:188 a posttranslational modification 157:188 a posttranslational modification of proteins 157:200 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 3 22 theme chemical 472:479 arg1 approaches 491:500 chemical digestion approaches 472:500 chemical digestion approaches 472:500 Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. 26132640 0 23 theme Quantitative 0:11 arg1 O-Glycomics 13:23 Quantitative O-Glycomics 0:23 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues 0:99 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues. 26132640 6 24 with Combined 1054:1061 arg1 extractions 1091:1101 sequential solid-phase extractions 1068:1101 sequential solid-phase extractions 1068:1101 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 2 25 theme known 422:426 arg1 endoglycosidase 428:442 no known endoglycosidase 419:442 no known endoglycosidase for such groups 419:458 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 0 26 theme Microwave-Assisted 28:45 arg1 β-Elimination 47:59 Microwave-Assisted β-Elimination 28:59 Microwave-Assisted β-Elimination 28:59 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues. 26132640 6 27 theme biological 1174:1183 arg1 samples 1185:1191 various biological samples 1166:1191 various biological samples 1166:1191 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 2 28 theme glycans 354:360 arg1 characterization 302:317 The characterization 298:317 The characterization of differently structured O-linked glycans (O-glycans) 298:372 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 2 28 theme glycans 354:360 arg1 challenging 390:400 challenging 390:400 challenging 390:400 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 6 29 theme sequential 1068:1077 arg1 extractions 1091:1101 sequential solid-phase extractions 1068:1101 sequential solid-phase extractions 1068:1101 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 30 theme O-glycomic 1143:1152 arg1 analyses 1154:1161 O-glycomic analyses 1143:1161 O-glycomic analyses of various biological samples 1143:1191 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 31 dep conditions 877:886 arg1 improved 932:939 improved 932:939 improved the recovery of total O-glycans from model glycoproteins (PSM) 932:1002 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 31 dep conditions 877:886 arg1 reduced 1030:1036 reduced 1030:1036 was reduced from 16 to 2 h. Combined with sequential solid-phase extractions 1026:1101 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 32 theme BEP 905:907 arg1 reaction 909:916 the MW-assisted BEP reaction 889:916 the MW-assisted BEP reaction 889:916 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 1 33 theme serine/threonine 128:143 arg1 residues 145:152 serine/threonine residues 128:152 serine/threonine residues 128:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 5 34 theme rapid 794:798 arg1 analysis 828:835 rapid and quantitative O-glycomic analysis 794:835 rapid and quantitative O-glycomic analysis 794:835 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 4 35 theme analogues 699:707 arg1 presence 676:683 the presence 672:683 the presence of pyrazolone analogues (BEP) 672:713 Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP). 26132640 1 36 theme protein 223:229 arg1 recognition 231:241 protein recognition 223:241 protein recognition 223:241 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 37 theme lipid 247:251 arg1 functions 253:261 lipid functions 247:261 lipid functions 247:261 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 3 38 used used 519:522 arg2 approaches 491:500 chemical digestion approaches 472:500 chemical digestion approaches 472:500 Therefore, chemical digestion approaches have been widely used; however, it is sometimes difficult to suppress unwanted side reactions. 26132640 4 39 theme pyrazolone 688:697 arg1 BEP 710:712 BEP 710:712 BEP 710:712 Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP). 26132640 4 39 theme pyrazolone 688:697 arg1 analogues 699:707 pyrazolone analogues 688:707 pyrazolone analogues (BEP) 688:713 Recently, we reported a novel O-glycomics procedure using β-elimination in the presence of pyrazolone analogues (BEP). 26132640 5 40 theme quantitative 804:815 arg1 analysis 828:835 rapid and quantitative O-glycomic analysis 794:835 rapid and quantitative O-glycomic analysis 794:835 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 2 41 link O-linked 345:352 arg1 glycans 354:360 differently structured O-linked glycans 322:360 differently structured O-linked glycans (O-glycans) 322:372 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 2 41 link O-linked 345:352 arg1 O-glycans 363:371 O-glycans 363:371 O-glycans 363:371 The characterization of differently structured O-linked glycans (O-glycans) is particularly challenging because there is no known endoglycosidase for such groups. 26132640 1 42 from functions 253:261 arg1 surfaces 271:278 cell surfaces 266:278 cell surfaces 266:278 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 43 theme residues 145:152 arg1 glycosylation 111:123 O-Linked glycosylation 102:123 O-Linked glycosylation of serine/threonine residues 102:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 43 theme residues 145:152 arg1 modification 177:188 a posttranslational modification 157:188 a posttranslational modification of proteins 157:200 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 5 44 theme O-glycomic 817:826 arg1 analysis 828:835 rapid and quantitative O-glycomic analysis 794:835 rapid and quantitative O-glycomic analysis 794:835 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 6 45 theme model 978:982 arg1 glycoproteins 984:996 model glycoproteins 978:996 model glycoproteins (PSM) 978:1002 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 45 theme model 978:982 arg1 PSM 999:1001 PSM 999:1001 PSM 999:1001 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 46 theme conditions 877:886 arg1 optimization 848:859 optimization 848:859 optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions 848:1101 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 1 47 from recognition 231:241 arg1 surfaces 271:278 cell surfaces 266:278 cell surfaces 266:278 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 5 48 theme present 723:729 arg1 study 731:735 the present study 719:735 the present study 719:735 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 6 49 theme reaction 868:875 arg1 conditions 877:886 the reaction conditions 864:886 the reaction conditions 864:886 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 50 theme samples 1185:1191 arg1 analyses 1154:1161 O-glycomic analyses 1143:1161 O-glycomic analyses of various biological samples 1143:1191 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 51 from glycoproteins 984:996 arg1 recovery 945:952 the recovery 941:952 the recovery of total O-glycans from model glycoproteins (PSM) 941:1002 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 6 52 theme solid-phase 1079:1089 arg1 extractions 1091:1101 sequential solid-phase extractions 1068:1101 sequential solid-phase extractions 1068:1101 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26132640 1 53 theme posttranslational 159:175 arg1 glycosylation 111:123 O-Linked glycosylation 102:123 O-Linked glycosylation of serine/threonine residues 102:152 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 53 theme posttranslational 159:175 arg1 modification 177:188 a posttranslational modification 157:188 a posttranslational modification of proteins 157:200 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 1 54 theme cell 266:269 arg1 surfaces 271:278 cell surfaces 266:278 cell surfaces 266:278 O-Linked glycosylation of serine/threonine residues is a posttranslational modification of proteins and is essential for protein recognition and lipid functions on cell surfaces and within cells. 26132640 0 55 theme Pyrazolone 80:89 arg1 Analogues 91:99 Pyrazolone Analogues 80:99 Pyrazolone Analogues 80:99 Quantitative O-Glycomics by Microwave-Assisted β-Elimination in the Presence of Pyrazolone Analogues. 26132640 5 56 theme BEP 776:778 arg1 procedure 780:788 a microwave (MW)-assisted BEP procedure 750:788 a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis 750:835 In the present study, we describe a microwave (MW)-assisted BEP procedure for rapid and quantitative O-glycomic analysis. 26132640 6 57 theme O-glycans 963:971 arg1 recovery 945:952 the recovery 941:952 the recovery of total O-glycans from model glycoproteins (PSM) 941:1002 Following optimization of the reaction conditions, the MW-assisted BEP reaction substantially improved the recovery of total O-glycans from model glycoproteins (PSM) and the reaction time was reduced from 16 to 2 h. Combined with sequential solid-phase extractions, this MW-assisted BEP procedure enabled O-glycomic analyses of various biological samples. 26858341 6 0 theme same 1084:1087 arg1 litters 1091:1097 the same 2 litters 1080:1097 the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)) 1080:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 2 1 theme pig 354:356 arg1 health 358:363 pig health 354:363 pig health 354:363 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 8 2 from pigs 1530:1533 arg1 abundant 1518:1525 abundant 1518:1525 abundant 1518:1525 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 2 3 theme interactions 284:295 arg1 understanding 227:239 An enhanced understanding 215:239 An enhanced understanding of infectious diseases and of host-microbe interactions 215:295 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 5 4 theme intestinal 855:864 arg1 mucosa 866:871 the intestinal mucosa 851:871 the intestinal mucosa 851:871 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 1 5 theme rising 171:176 arg1 resistance 188:197 the rapidly rising bacterial resistance 159:197 the rapidly rising bacterial resistance to antibiotics 159:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 5 6 theme O-Linked 831:838 arg1 glycans 840:846 O-Linked glycans 831:846 O-Linked glycans 831:846 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 1 7 theme industrial 113:122 arg1 production 128:137 industrial pig production 113:137 industrial pig production 113:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 2 8 theme enhanced 218:225 arg1 understanding 227:239 An enhanced understanding 215:239 An enhanced understanding of infectious diseases and of host-microbe interactions 215:295 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 4 9 theme fimbriated 691:700 arg1 pathogen 678:685 the common porcine enteric pathogen F18 651:689 the common porcine enteric pathogen F18 fimbriated Escherichia coli 651:717 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 8 10 with pigs 1530:1533 arg1 mutation 1558:1565 the FUT1 missense mutation 1540:1565 the FUT1 missense mutation 1540:1565 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 8 11 gly sialylated 1475:1484 arg1 structures 1486:1495 sialylated structures 1475:1495 sialylated structures 1475:1495 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 9 12 theme host-microbe 1713:1724 arg1 interactions 1726:1737 host-microbe interactions 1713:1737 host-microbe interactions 1713:1737 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 2 13 theme diseases 255:262 arg1 understanding 227:239 An enhanced understanding 215:239 An enhanced understanding of infectious diseases and of host-microbe interactions 215:295 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 8 14 theme H-antigen 1450:1458 arg1 precursors 1460:1469 H-antigen precursors 1450:1469 H-antigen precursors 1450:1469 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 7 15 theme glycans 1290:1296 arg1 glycans 1290:1296 the O-linked glycans 1277:1296 the O-linked glycans 1277:1296 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 7 15 theme glycans 1290:1296 arg1 11 1271:1272 11 1271:1272 11 1271:1272 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 4 16 theme porcine 662:668 arg1 pathogen 678:685 the common porcine enteric pathogen F18 651:689 the common porcine enteric pathogen F18 fimbriated Escherichia coli 651:717 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 7 17 theme glycans 1200:1206 arg1 None 1162:1165 None 1162:1165 None of the characterized 48 N-linked glycans 1162:1206 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 5 18 theme N- 824:825 arg1 proteins 873:880 N- and O-Linked glycans of the intestinal mucosa proteins 824:880 N- and O-Linked glycans of the intestinal mucosa proteins 824:880 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 4 19 theme G 508:508 arg1 mutation 540:547 a G > A (nucleotide 307) missense mutation 506:547 a G > A (nucleotide 307) missense mutation 506:547 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 6 20 theme heterozygous 1004:1015 arg1 FUT1-307 1023:1030 FUT1-307 1023:1030 FUT1-307 1023:1030 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 20 theme heterozygous 1004:1015 arg1 pigs 1017:1020 four heterozygous pigs 999:1020 four heterozygous pigs (FUT1-307(A/G)) 999:1036 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 4 21 theme A 512:512 arg1 mutation 540:547 a G > A (nucleotide 307) missense mutation 506:547 a G > A (nucleotide 307) missense mutation 506:547 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 9 22 from variation 1682:1690 arg1 FUT1 1695:1698 FUT1 1695:1698 FUT1 1695:1698 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 9 23 theme genetic 1766:1772 arg1 variant 1774:1780 the FUT1 genetic variant 1757:1780 the FUT1 genetic variant 1757:1780 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 4 24 theme glycan 759:764 arg1 structures 766:775 glycan structures 759:775 glycan structures potentially involved in intestinal infections 759:821 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 7 25 theme O-linked 1281:1288 arg1 glycans 1290:1296 the O-linked glycans 1277:1296 the O-linked glycans 1277:1296 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 0 26 theme mucosa 73:78 arg1 glycosylation 37:49 impact protein glycosylation 22:49 impact protein glycosylation of porcine intestinal mucosa 22:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 0 27 theme FUT1 0:3 arg1 variants 13:20 FUT1 genetic variants 0:20 FUT1 genetic variants 0:20 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 4 28 dep Escherichia 702:712 arg1 coli 714:717 coli 714:717 coli 714:717 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 6 29 theme age-matched 1047:1057 arg1 pigs 1070:1073 four age-matched homozygous pigs 1042:1073 four age-matched homozygous pigs 1042:1073 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 4 30 theme unique 731:736 arg1 opportunity 738:748 a unique opportunity 729:748 a unique opportunity to study glycan structures potentially involved in intestinal infections 729:821 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 4 31 from mutation 540:547 arg1 gene 589:592 porcine α1,2fucosyltransferase 1 gene 556:592 the porcine α1,2fucosyltransferase 1 gene (FUT1) 552:599 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 4 31 from mutation 540:547 arg1 FUT1 595:598 FUT1 595:598 FUT1 595:598 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 1 32 theme antibiotics 98:108 arg1 use 91:93 A massive use 81:93 A massive use of antibiotics in industrial pig production 81:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 1 32 theme antibiotics 98:108 arg1 cause 150:154 a major cause 142:154 a major cause of the rapidly rising bacterial resistance to antibiotics 142:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 9 33 theme pig 1802:1804 arg1 health 1810:1815 pig gut health 1802:1815 pig gut health 1802:1815 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 3 34 theme microbe-carrying 465:480 arg1 lectins 482:488 microbe-carrying lectins 465:488 microbe-carrying lectins 465:488 Host-microbe interactions depend on host-expressed glycans and microbe-carrying lectins. 26858341 8 35 from abundant 1518:1525 arg1 pigs 1530:1533 pigs 1530:1533 pigs with the FUT1 missense mutation 1530:1565 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 0 36 theme protein 29:35 arg1 glycosylation 37:49 impact protein glycosylation 22:49 impact protein glycosylation of porcine intestinal mucosa 22:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 4 37 theme nucleotide 515:524 arg1 mutation 540:547 a G > A (nucleotide 307) missense mutation 506:547 a G > A (nucleotide 307) missense mutation 506:547 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 0 38 theme intestinal 62:71 arg1 mucosa 73:78 porcine intestinal mucosa 54:78 porcine intestinal mucosa 54:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 8 39 theme overall 1369:1375 arg1 abundance 1377:1385 The overall abundance 1365:1385 The overall abundance of H-antigen carrying structures 1365:1418 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 4 40 theme > 510:510 arg1 mutation 540:547 a G > A (nucleotide 307) missense mutation 506:547 a G > A (nucleotide 307) missense mutation 506:547 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 6 41 theme missense 1112:1119 arg1 FUT1-307 1146:1153 FUT1-307 1146:1153 FUT1-307(A/A) 1146:1158 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 41 theme missense 1112:1119 arg1 constellation 1131:1143 the missense FUT1 gene constellation 1108:1143 the missense FUT1 gene constellation (FUT1-307(A/A)) 1108:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 0 42 gly glycosylation 37:49 arg1 mucosa 73:78 porcine intestinal mucosa 54:78 porcine intestinal mucosa 54:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 1 43 from use 91:93 arg1 production 128:137 industrial pig production 113:137 industrial pig production 113:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 6 44 theme Relative 927:934 arg1 abundances 936:945 Relative abundances 927:945 Relative abundances of all glycans 927:960 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 9 45 theme FUT1 1615:1618 arg1 role 1607:1610 the role 1603:1610 the role of FUT1 on intestinal glycosylation 1603:1646 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 8 46 theme missense 1549:1556 arg1 mutation 1558:1565 the FUT1 missense mutation 1540:1565 the FUT1 missense mutation 1540:1565 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 9 47 theme intestinal 1623:1632 arg1 glycosylation 1634:1646 intestinal glycosylation 1623:1646 intestinal glycosylation 1623:1646 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 4 48 theme α1,2fucosyltransferase 564:585 arg1 gene 589:592 porcine α1,2fucosyltransferase 1 gene 556:592 the porcine α1,2fucosyltransferase 1 gene (FUT1) 552:599 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 4 48 theme α1,2fucosyltransferase 564:585 arg1 FUT1 595:598 FUT1 595:598 FUT1 595:598 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 2 49 theme host-microbe 271:282 arg1 interactions 284:295 host-microbe interactions 271:295 host-microbe interactions 271:295 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 5 50 theme mucosa 866:871 arg1 glycans 840:846 O-Linked glycans 831:846 O-Linked glycans 831:846 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 5 50 theme mucosa 866:871 arg1 N- 824:825 N- 824:825 N- 824:825 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 7 51 link O-linked 1281:1288 arg1 glycans 1290:1296 the O-linked glycans 1277:1296 the O-linked glycans 1277:1296 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 7 52 link N-linked 1191:1198 arg1 glycans 1200:1206 the characterized 48 N-linked glycans 1170:1206 the characterized 48 N-linked glycans 1170:1206 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 1 53 theme bacterial 178:186 arg1 resistance 188:197 the rapidly rising bacterial resistance 159:197 the rapidly rising bacterial resistance to antibiotics 159:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 6 54 contain carrying 1099:1106 arg1 litters 1091:1097 the same 2 litters 1080:1097 the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)) 1080:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 54 contain carrying 1099:1106 arg2 FUT1-307 1146:1153 FUT1-307 1146:1153 FUT1-307(A/A) 1146:1158 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 54 contain carrying 1099:1106 arg2 constellation 1131:1143 the missense FUT1 gene constellation 1108:1143 the missense FUT1 gene constellation (FUT1-307(A/A)) 1108:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 55 theme gene 1126:1129 arg1 FUT1-307 1146:1153 FUT1-307 1146:1153 FUT1-307(A/A) 1146:1158 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 55 theme gene 1126:1129 arg1 constellation 1131:1143 the missense FUT1 gene constellation 1108:1143 the missense FUT1 gene constellation (FUT1-307(A/A)) 1108:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 8 56 theme FUT1 1544:1547 arg1 mutation 1558:1565 the FUT1 missense mutation 1540:1565 the FUT1 missense mutation 1540:1565 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 7 57 theme FUT1 1241:1244 arg1 mutation 1255:1262 the FUT1 missense mutation 1237:1262 the FUT1 missense mutation 1237:1262 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 5 58 theme glycans 840:846 arg1 proteins 873:880 N- and O-Linked glycans of the intestinal mucosa proteins 824:880 N- and O-Linked glycans of the intestinal mucosa proteins 824:880 N- and O-Linked glycans of the intestinal mucosa proteins were characterized in detail using LC-MS/MS. 26858341 6 59 theme glycans 954:960 arg1 abundances 936:945 Relative abundances 927:945 Relative abundances of all glycans 927:960 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 1 60 theme pig 124:126 arg1 production 128:137 industrial pig production 113:137 industrial pig production 113:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 7 61 theme altered 1319:1325 arg1 abundances 1327:1336 significantly altered abundances 1305:1336 significantly altered abundances between the two genotypes 1305:1362 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 7 62 theme N-linked 1191:1198 arg1 glycans 1200:1206 the characterized 48 N-linked glycans 1170:1206 the characterized 48 N-linked glycans 1170:1206 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 2 63 theme infectious 244:253 arg1 diseases 255:262 infectious diseases 244:262 infectious diseases 244:262 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 8 64 theme sialylated 1475:1484 arg1 structures 1486:1495 sialylated structures 1475:1495 sialylated structures 1475:1495 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 4 65 theme common 655:660 arg1 pathogen 678:685 the common porcine enteric pathogen F18 651:689 the common porcine enteric pathogen F18 fimbriated Escherichia coli 651:717 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 7 66 theme characterized 1174:1186 arg1 glycans 1200:1206 the characterized 48 N-linked glycans 1170:1206 the characterized 48 N-linked glycans 1170:1206 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 1 67 theme major 144:148 arg1 use 91:93 A massive use 81:93 A massive use of antibiotics in industrial pig production 81:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 1 67 theme major 144:148 arg1 cause 150:154 a major cause 142:154 a major cause of the rapidly rising bacterial resistance to antibiotics 142:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 4 68 theme enteric 670:676 arg1 pathogen 678:685 the common porcine enteric pathogen F18 651:689 the common porcine enteric pathogen F18 fimbriated Escherichia coli 651:717 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 9 69 theme FUT1 1761:1764 arg1 variant 1774:1780 the FUT1 genetic variant 1757:1780 the FUT1 genetic variant 1757:1780 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 1 70 theme resistance 188:197 arg1 use 91:93 A massive use 81:93 A massive use of antibiotics in industrial pig production 81:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 1 70 theme resistance 188:197 arg1 cause 150:154 a major cause 142:154 a major cause of the rapidly rising bacterial resistance to antibiotics 142:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 0 71 theme genetic 5:11 arg1 variants 13:20 FUT1 genetic variants 0:20 FUT1 genetic variants 0:20 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 1 72 theme massive 83:89 arg1 use 91:93 A massive use 81:93 A massive use of antibiotics in industrial pig production 81:137 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 1 72 theme massive 83:89 arg1 cause 150:154 a major cause 142:154 a major cause of the rapidly rising bacterial resistance to antibiotics 142:212 A massive use of antibiotics in industrial pig production is a major cause of the rapidly rising bacterial resistance to antibiotics. 26858341 9 73 from role 1607:1610 arg1 glycosylation 1634:1646 intestinal glycosylation 1623:1646 intestinal glycosylation 1623:1646 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 6 74 theme homozygous 1059:1068 arg1 pigs 1070:1073 four age-matched homozygous pigs 1042:1073 four age-matched homozygous pigs 1042:1073 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 8 75 contain carrying 1400:1407 arg2 structures 1409:1418 structures 1409:1418 structures 1409:1418 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 8 75 contain carrying 1400:1407 arg1 H-antigen 1390:1398 H-antigen 1390:1398 H-antigen carrying structures 1390:1418 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 3 76 theme host-expressed 438:451 arg1 glycans 453:459 host-expressed glycans 438:459 host-expressed glycans 438:459 Host-microbe interactions depend on host-expressed glycans and microbe-carrying lectins. 26858341 0 77 theme impact 22:27 arg1 glycosylation 37:49 impact protein glycosylation 22:49 impact protein glycosylation of porcine intestinal mucosa 22:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 4 78 dep fimbriated 691:700 arg1 Escherichia 702:712 Escherichia 702:712 Escherichia 702:712 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 4 79 theme porcine 556:562 arg1 gene 589:592 porcine α1,2fucosyltransferase 1 gene 556:592 the porcine α1,2fucosyltransferase 1 gene (FUT1) 552:599 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 4 79 theme porcine 556:562 arg1 FUT1 595:598 FUT1 595:598 FUT1 595:598 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 2 80 contain has 297:299 arg2 potential 305:313 the potential to explore alternative ways 301:341 the potential to explore alternative ways 301:341 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 2 80 contain has 297:299 arg1 understanding 227:239 An enhanced understanding 215:239 An enhanced understanding of infectious diseases and of host-microbe interactions 215:295 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 7 81 theme missense 1246:1253 arg1 mutation 1255:1262 the FUT1 missense mutation 1237:1262 the FUT1 missense mutation 1237:1262 None of the characterized 48 N-linked glycans was found to be regulated by the FUT1 missense mutation, while 11 of the O-linked glycans showed significantly altered abundances between the two genotypes. 26858341 9 82 theme gut 1806:1808 arg1 health 1810:1815 pig gut health 1802:1815 pig gut health 1802:1815 These results provide insight into the role of FUT1 on intestinal glycosylation, improve our understanding of how variation in FUT1 can modulate host-microbe interactions, and suggest that the FUT1 genetic variant may help to improve pig gut health. 26858341 4 83 theme intestinal 801:810 arg1 infections 812:821 intestinal infections 801:821 intestinal infections 801:821 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 0 84 theme porcine 54:60 arg1 mucosa 73:78 porcine intestinal mucosa 54:78 porcine intestinal mucosa 54:78 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 0 85 dep glycosylation 37:49 arg1 variants 13:20 FUT1 genetic variants 0:20 FUT1 genetic variants 0:20 FUT1 genetic variants impact protein glycosylation of porcine intestinal mucosa. 26858341 6 86 theme FUT1 1121:1124 arg1 FUT1-307 1146:1153 FUT1-307 1146:1153 FUT1-307(A/A) 1146:1158 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 86 theme FUT1 1121:1124 arg1 constellation 1131:1143 the missense FUT1 gene constellation 1108:1143 the missense FUT1 gene constellation (FUT1-307(A/A)) 1108:1159 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 3 87 theme Host-microbe 402:413 arg1 interactions 415:426 Host-microbe interactions 402:426 Host-microbe interactions 402:426 Host-microbe interactions depend on host-expressed glycans and microbe-carrying lectins. 26858341 4 88 theme missense 531:538 arg1 mutation 540:547 a G > A (nucleotide 307) missense mutation 506:547 a G > A (nucleotide 307) missense mutation 506:547 In this study, a G > A (nucleotide 307) missense mutation in the porcine α1,2fucosyltransferase 1 gene (FUT1), which has been reported to prevent infections by the common porcine enteric pathogen F18 fimbriated Escherichia coli, provided a unique opportunity to study glycan structures potentially involved in intestinal infections. 26858341 2 89 theme alternative 326:336 arg1 ways 338:341 alternative ways 326:341 alternative ways 326:341 An enhanced understanding of infectious diseases and of host-microbe interactions has the potential to explore alternative ways to improve pig health and reduce the need for antibiotics. 26858341 8 90 theme H-antigen 1390:1398 arg1 abundance 1377:1385 The overall abundance 1365:1385 The overall abundance of H-antigen carrying structures 1365:1418 The overall abundance of H-antigen carrying structures was decreased fivefold, while H-antigen precursors and sialylated structures were relatively more abundant in pigs with the FUT1 missense mutation. 26858341 6 91 from litters 1091:1097 arg1 FUT1-307 1023:1030 FUT1-307 1023:1030 FUT1-307 1023:1030 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 91 from litters 1091:1097 arg1 pigs 1017:1020 four heterozygous pigs 999:1020 four heterozygous pigs (FUT1-307(A/G)) 999:1036 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 26858341 6 91 from litters 1091:1097 arg1 pigs 1070:1073 four age-matched homozygous pigs 1042:1073 four age-matched homozygous pigs 1042:1073 Relative abundances of all glycans were determined and compared between four heterozygous pigs (FUT1-307(A/G)) and four age-matched homozygous pigs from the same 2 litters carrying the missense FUT1 gene constellation (FUT1-307(A/A)). 27694851 7 0 theme certain 1215:1221 arg1 motifs 1231:1236 certain peptide motifs 1215:1236 certain peptide motifs 1215:1236 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 2 1 theme unique 380:385 arg1 sites 395:399 unique binding sites 380:399 unique binding sites for specific ligands 380:420 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 7 2 theme peptide 1223:1229 arg1 motifs 1231:1236 certain peptide motifs 1215:1236 certain peptide motifs 1215:1236 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 0 3 from identification 14:27 arg1 proteoglycans 93:105 hybrid proteoglycans 86:105 hybrid proteoglycans 86:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 8 4 theme basic 1372:1376 arg1 biology 1378:1384 basic biology 1372:1384 basic biology 1372:1384 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 4 5 theme glycoproteomic 604:617 arg1 approach 619:626 a glycoproteomic approach 602:626 a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins 602:739 Here we present a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins. 27694851 8 6 gly glycosylations 1452:1465 arg1 proteoglycans 1470:1482 proteoglycans 1470:1482 proteoglycans 1470:1482 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 7 7 from locus 1194:1198 arg1 Inspection 1131:1140 Inspection 1131:1140 Inspection of the amino acid sequence at the hybrid attachment locus 1131:1198 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 5 8 dep HS 929:930 arg1 lengths 945:951 chain lengths 939:951 chain lengths 939:951 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 0 9 from glycosaminoglycans 64:81 arg1 proteoglycans 93:105 hybrid proteoglycans 86:105 hybrid proteoglycans 86:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 7 10 theme amino 1149:1153 arg1 sequence 1160:1167 the amino acid sequence 1145:1167 the amino acid sequence 1145:1167 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 8 11 theme fundamental 1347:1357 arg1 questions 1359:1367 fundamental questions 1347:1367 fundamental questions in basic biology 1347:1384 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 6 12 theme CS 1121:1122 arg1 chain 1124:1128 either a HS or a CS chain 1104:1128 chain 1124:1128 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 7 13 theme attachment 1183:1192 arg1 locus 1194:1198 the hybrid attachment locus 1172:1198 the hybrid attachment locus 1172:1198 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 2 14 theme opposite 301:308 arg1 effects 310:316 opposite effects 301:316 opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands 301:420 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 1 15 theme complex 162:168 arg1 sulfate 116:122 Heparan sulfate 108:122 Heparan sulfate (HS) 108:127 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 15 theme complex 162:168 arg1 sulfate 145:151 chondroitin sulfate 133:151 chondroitin sulfate (CS) 133:156 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 15 theme complex 162:168 arg1 polysaccharides 170:184 complex polysaccharides 162:184 complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms 162:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 5 16 theme complex 787:793 arg1 perlecan 775:782 perlecan 775:782 perlecan 775:782 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 5 16 theme complex 787:793 arg1 proteoglycan 795:806 a complex proteoglycan 785:806 a complex proteoglycan known to be substituted with both HS and CS chains 785:857 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 5 17 dep HS 842:843 arg1 chains 852:857 chains 852:857 chains 852:857 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 6 18 theme consensus 1055:1063 arg1 sites 1068:1072 three consensus HS sites 1049:1072 three consensus HS sites 1049:1072 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 2 19 dep HS 341:342 arg1 motifs 362:367 structural motifs 351:367 structural motifs 351:367 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 1 20 theme Heparan 108:114 arg1 sulfate 116:122 Heparan sulfate 108:122 Heparan sulfate (HS) 108:127 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 20 theme Heparan 108:114 arg1 sulfate 145:151 chondroitin sulfate 133:151 chondroitin sulfate (CS) 133:156 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 20 theme Heparan 108:114 arg1 polysaccharides 170:184 complex polysaccharides 162:184 complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms 162:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 20 theme Heparan 108:114 arg1 HS 125:126 HS 125:126 HS 125:126 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 7 21 theme selection 1268:1276 arg1 process 1278:1284 the chain type selection process 1253:1284 the chain type selection process 1253:1284 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 2 22 theme specific 405:412 arg1 ligands 414:420 specific ligands 405:420 specific ligands 405:420 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 0 23 theme Site-specific 0:12 arg1 identification 14:27 Site-specific identification 0:27 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 0:106 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 5 24 theme chondroitinase 896:909 arg1 ABC 911:913 chondroitinase ABC 896:913 chondroitinase ABC 896:913 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 6 25 theme site 1089:1092 arg1 identification 1031:1044 the identification 1027:1044 the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain 1027:1128 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 0 26 theme heparan 32:38 arg1 glycosaminoglycans 64:81 heparan and chondroitin sulfate glycosaminoglycans 32:81 heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans 32:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 8 27 theme proteoglycans 1470:1482 arg1 glycosylations 1452:1465 site-specific glycosylations 1438:1465 site-specific glycosylations of proteoglycans 1438:1482 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 1 28 theme important 200:208 arg1 pathways 221:228 important biological pathways 200:228 important biological pathways in virtually all metazoan organisms 200:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 3 29 theme physiology 559:568 arg1 regulation 535:544 the regulation 531:544 the regulation of mammalian physiology 531:568 Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. 27694851 2 30 theme cell 321:324 arg1 functions 326:334 cell functions 321:334 cell functions 321:334 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 1 31 theme biological 210:219 arg1 pathways 221:228 important biological pathways 200:228 important biological pathways in virtually all metazoan organisms 200:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 0 32 theme chondroitin 44:54 arg1 glycosaminoglycans 64:81 heparan and chondroitin sulfate glycosaminoglycans 32:81 heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans 32:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 5 33 theme chain 939:943 arg1 lengths 945:951 chain lengths 939:951 chain lengths 939:951 Glycopeptides were prepared from perlecan, a complex proteoglycan known to be substituted with both HS and CS chains, further digested with heparinase or chondroitinase ABC to reduce the HS and CS chain lengths respectively, and thereafter analyzed by nLC-MS/MS. 27694851 0 34 from proteoglycans 93:105 arg1 identification 14:27 Site-specific identification 0:27 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 0:106 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 8 35 theme functional 1418:1427 arg1 roles 1429:1433 the functional roles 1414:1433 the functional roles of site-specific glycosylations of proteoglycans 1414:1482 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 3 36 theme glycan 454:459 arg1 biosynthesis 461:472 glycan biosynthesis 454:472 glycan biosynthesis 454:472 Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. 27694851 7 37 theme acid 1155:1158 arg1 sequence 1160:1167 the amino acid sequence 1145:1167 the amino acid sequence 1145:1167 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 7 38 theme sequence 1160:1167 arg1 Inspection 1131:1140 Inspection 1131:1140 Inspection of the amino acid sequence at the hybrid attachment locus 1131:1198 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 1 39 from pathways 221:228 arg1 organisms 256:264 virtually all metazoan organisms 233:264 virtually all metazoan organisms 233:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 3 40 theme complex 484:490 arg1 HS 492:493 complex HS 484:493 complex HS 484:493 Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. 27694851 0 41 theme glycosaminoglycans 64:81 arg1 identification 14:27 Site-specific identification 0:27 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 0:106 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 6 42 theme sites 1068:1072 arg1 identification 1031:1044 the identification 1027:1044 the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain 1027:1128 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 6 43 theme a 1119:1119 arg1 chain 1124:1128 either a HS or a CS chain 1104:1128 chain 1124:1128 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 8 44 theme analytical 1292:1301 arg1 approach 1303:1310 This analytical approach 1287:1310 This analytical approach 1287:1310 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 0 45 theme sulfate 56:62 arg1 glycosaminoglycans 64:81 heparan and chondroitin sulfate glycosaminoglycans 32:81 heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans 32:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 4 46 dep HS 671:672 arg1 sites 692:696 attachment sites 681:696 attachment sites 681:696 Here we present a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins. 27694851 2 47 theme structural 351:360 arg1 motifs 362:367 structural motifs 351:367 structural motifs 351:367 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 7 48 theme chain 1257:1261 arg1 process 1278:1284 the chain type selection process 1253:1284 the chain type selection process 1253:1284 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 7 49 theme hybrid 1176:1181 arg1 locus 1194:1198 the hybrid attachment locus 1172:1198 the hybrid attachment locus 1172:1198 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 8 50 theme glycosylations 1452:1465 arg1 roles 1429:1433 the functional roles 1414:1433 the functional roles of site-specific glycosylations of proteoglycans 1414:1482 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 1 51 theme chondroitin 133:143 arg1 CS 154:155 CS 154:155 CS 154:155 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 51 theme chondroitin 133:143 arg1 sulfate 145:151 chondroitin sulfate 133:151 chondroitin sulfate (CS) 133:156 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 51 theme chondroitin 133:143 arg1 polysaccharides 170:184 complex polysaccharides 162:184 complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms 162:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 1 51 theme chondroitin 133:143 arg1 sulfate 116:122 Heparan sulfate 108:122 Heparan sulfate (HS) 108:127 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 6 52 theme HS 1065:1066 arg1 sites 1068:1072 three consensus HS sites 1049:1072 three consensus HS sites 1049:1072 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 8 53 theme site-specific 1438:1450 arg1 glycosylations 1452:1465 site-specific glycosylations 1438:1465 site-specific glycosylations of proteoglycans 1438:1482 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 3 54 theme mammalian 549:557 arg1 physiology 559:568 mammalian physiology 549:568 mammalian physiology 549:568 Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. 27694851 8 55 from questions 1359:1367 arg1 biology 1378:1384 basic biology 1372:1384 basic biology 1372:1384 This analytical approach will become useful when addressing fundamental questions in basic biology specifically in elucidating the functional roles of site-specific glycosylations of proteoglycans. 27694851 1 56 theme all 243:245 arg1 organisms 256:264 virtually all metazoan organisms 233:264 virtually all metazoan organisms 233:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 0 57 theme hybrid 86:91 arg1 proteoglycans 93:105 hybrid proteoglycans 86:105 hybrid proteoglycans 86:105 Site-specific identification of heparan and chondroitin sulfate glycosaminoglycans in hybrid proteoglycans. 27694851 4 58 theme attachment 681:690 arg1 sites 692:696 attachment sites 681:696 attachment sites 681:696 Here we present a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins. 27694851 3 59 dep HS 492:493 arg1 polysaccharides 502:516 polysaccharides 502:516 polysaccharides 502:516 Still, the mechanisms by which glycan biosynthesis generates complex HS and CS polysaccharides required for the regulation of mammalian physiology remain elusive. 27694851 6 60 theme hybrid 1082:1087 arg1 site 1089:1092 one hybrid site 1078:1092 one hybrid site 1078:1092 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 4 61 theme core 727:730 arg1 proteins 732:739 the core proteins 723:739 the core proteins 723:739 Here we present a glycoproteomic approach that identifies and differentiates between HS and CS attachment sites and provides identity to the core proteins. 27694851 2 62 from effects 310:316 arg1 functions 326:334 cell functions 321:334 cell functions 321:334 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 1 63 theme metazoan 247:254 arg1 organisms 256:264 virtually all metazoan organisms 233:264 virtually all metazoan organisms 233:264 Heparan sulfate (HS) and chondroitin sulfate (CS) are complex polysaccharides that regulate important biological pathways in virtually all metazoan organisms. 27694851 7 64 theme type 1263:1266 arg1 process 1278:1284 the chain type selection process 1253:1284 the chain type selection process 1253:1284 Inspection of the amino acid sequence at the hybrid attachment locus indicates that certain peptide motifs may encode for the chain type selection process. 27694851 2 65 theme binding 387:393 arg1 sites 395:399 unique binding sites 380:399 unique binding sites for specific ligands 380:420 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 6 66 contain carrying 1095:1102 arg1 sites 1068:1072 three consensus HS sites 1049:1072 three consensus HS sites 1049:1072 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 6 66 contain carrying 1095:1102 arg2 chain 1124:1128 either a HS or a CS chain 1104:1128 chain 1124:1128 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 6 66 contain carrying 1095:1102 arg2 HS 1113:1114 either a HS or a CS chain 1104:1128 HS 1113:1114 This protocol enabled the identification of three consensus HS sites and one hybrid site, carrying either a HS or a CS chain. 27694851 2 67 with effects 310:316 arg1 CS 348:349 CS 348:349 CS 348:349 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 27694851 2 67 with effects 310:316 arg1 HS 341:342 HS 341:342 HS 341:342 The polysaccharides often display opposite effects on cell functions with HS and CS structural motifs presenting unique binding sites for specific ligands. 25833378 8 0 dep 11-fold 1215:1221 arg1 to 1212:1213 to 1212:1213 to 1212:1213 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 7 1 theme three-dimensional 873:889 arg1 model 900:904 A three-dimensional homology model 871:904 A three-dimensional homology model 871:904 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 2 theme site 1134:1137 arg1 residue 1139:1145 an active site residue 1124:1145 an active site residue 1124:1145 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 5 3 dep Salmonella 704:713 arg1 enterica 715:722 enterica 715:722 enterica 715:722 In this study, we increased glycosylation rates for Salmonella enterica sv. 25833378 9 4 theme mutant 1360:1365 arg1 S80R-Q287P-N311V 1367:1382 a triple mutant S80R-Q287P-N311V 1351:1382 a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16 1351:1464 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 7 5 theme acceptor 994:1001 arg1 peptide 1003:1009 the acceptor peptide 990:1009 the acceptor peptide 990:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 6 theme enhancing 1159:1167 arg1 N311V 1178:1182 the enhancing mutation N311V 1155:1182 the enhancing mutation N311V 1155:1182 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 6 7 theme LT2 740:742 arg1 antigen 746:752 Typhimurium LT2 O antigen 728:752 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) 728:782 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 9 8 theme yield 1395:1399 arg1 improvement 1401:1411 a yield improvement 1393:1411 a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16 1393:1464 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 2 9 theme glycoconjugate 371:384 arg1 vaccines 386:393 glycoconjugate vaccines 371:393 glycoconjugate vaccines 371:393 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 7 10 attach attached 978:985 arg2 N-glycan 969:976 the natural C. jejuni N-glycan 947:976 the natural C. jejuni N-glycan attached to the acceptor peptide 947:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 7 10 attach attached 978:985 arg1 peptide 1003:1009 the acceptor peptide 990:1009 the acceptor peptide 990:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 4 11 theme Transfer 563:570 arg1 efficiency 572:581 Transfer efficiency 563:581 Transfer efficiency of PglBCj 563:591 Transfer efficiency of PglBCj is low for certain heterologous polysaccharide substrates. 25833378 9 12 theme enterica 1419:1426 arg1 LT2 1428:1430 S. enterica LT2 1416:1430 S. enterica LT2 glycoconjugates 1416:1446 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 2 13 theme Campylobacter 295:307 arg1 jejuni 309:314 Campylobacter jejuni 295:314 Campylobacter jejuni 295:314 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 9 14 theme glycoconjugates 1432:1446 arg1 improvement 1401:1411 a yield improvement 1393:1411 a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16 1393:1464 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 2 15 theme PglB 287:290 arg1 expression 237:246 Functional expression 226:246 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli 226:334 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 5 16 theme glycosylation 680:692 arg1 rates 694:698 glycosylation rates 680:698 glycosylation rates for Salmonella enterica sv 680:725 In this study, we increased glycosylation rates for Salmonella enterica sv. 25833378 1 17 theme efficient 149:157 arg1 measures 170:177 the most efficient preventive measures 140:177 the most efficient preventive measures against life-threatening bacterial infections 140:223 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 10 18 theme protein 1586:1592 arg1 engineering 1594:1604 structure-guided protein engineering 1569:1604 structure-guided protein engineering 1569:1604 Our results demonstrate that bacterial N-OST can be tailored to specific polysaccharide substrates by structure-guided protein engineering. 25833378 3 19 theme Polysaccharide 417:430 arg1 antigens 432:439 Polysaccharide antigens 417:439 Polysaccharide antigens of pathogenic bacteria 417:462 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 6 20 theme N-acetyl 767:774 arg1 sugars 776:781 N-acetyl sugars 767:781 N-acetyl sugars 767:781 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 0 21 theme structure-guided 81:96 arg1 engineering 98:108 structure-guided engineering 81:108 structure-guided engineering 81:108 Increased efficiency of Campylobacter jejuni N-oligosaccharyltransferase PglB by structure-guided engineering. 25833378 1 22 theme life-threatening 187:202 arg1 infections 214:223 life-threatening bacterial infections 187:223 life-threatening bacterial infections 187:223 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 4 23 theme heterologous 612:623 arg1 substrates 640:649 certain heterologous polysaccharide substrates 604:649 certain heterologous polysaccharide substrates 604:649 Transfer efficiency of PglBCj is low for certain heterologous polysaccharide substrates. 25833378 3 24 theme bacteria 455:462 arg1 antigens 432:439 Polysaccharide antigens 417:439 Polysaccharide antigens of pathogenic bacteria 417:462 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 10 25 theme structure-guided 1569:1584 arg1 engineering 1594:1604 structure-guided protein engineering 1569:1604 structure-guided protein engineering 1569:1604 Our results demonstrate that bacterial N-OST can be tailored to specific polysaccharide substrates by structure-guided protein engineering. 25833378 1 26 theme Conjugate 111:119 arg1 vaccines 121:128 Conjugate vaccines 111:128 Conjugate vaccines 111:128 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 3 27 theme engineered 531:540 arg1 sites 556:560 engineered glycosylation sites 531:560 engineered glycosylation sites 531:560 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 0 28 theme Increased 0:8 arg1 efficiency 10:19 Increased efficiency 0:19 Increased efficiency of Campylobacter jejuni 0:43 Increased efficiency of Campylobacter jejuni N-oligosaccharyltransferase PglB by structure-guided engineering. 25833378 7 29 theme C. 959:960 arg1 jejuni 962:967 natural C. jejuni 951:967 the natural C. jejuni N-glycan attached to the acceptor peptide 947:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 30 theme in 1233:1234 arg1 rates 1255:1259 in vivo glycosylation rates 1233:1259 in vivo glycosylation rates 1233:1259 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 8 31 theme glycosylation 1241:1253 arg1 rates 1255:1259 in vivo glycosylation rates 1233:1259 in vivo glycosylation rates 1233:1259 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 6 32 theme Staphylococcus 788:801 arg1 CP5 810:812 Staphylococcus aureus CP5 788:812 Staphylococcus aureus CP5 788:812 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 0 33 theme jejuni 38:43 arg1 efficiency 10:19 Increased efficiency 0:19 Increased efficiency of Campylobacter jejuni 0:43 Increased efficiency of Campylobacter jejuni N-oligosaccharyltransferase PglB by structure-guided engineering. 25833378 3 34 theme immunogenic 493:503 arg1 proteins 514:521 immunogenic acceptor proteins 493:521 immunogenic acceptor proteins bearing engineered glycosylation sites 493:560 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 7 35 theme PglBCj 929:934 arg1 model 900:904 A three-dimensional homology model 871:904 A three-dimensional homology model 871:904 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 36 theme Saturation 1098:1107 arg1 mutagenesis 1109:1119 Saturation mutagenesis 1098:1119 Saturation mutagenesis of an active site residue 1098:1145 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 2 37 theme N-OST 280:284 arg1 PglB 287:290 N-oligosaccharyltransferase (N-OST) PglB 251:290 N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni 251:314 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 9 38 theme in 1325:1326 arg1 evolution 1334:1342 in vitro evolution 1325:1342 in vitro evolution 1325:1342 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 2 39 theme N-oligosaccharyltransferase 251:277 arg1 PglB 287:290 N-oligosaccharyltransferase (N-OST) PglB 251:290 N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni 251:314 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 3 40 gly glycosylation 542:554 arg2 sites 556:560 engineered glycosylation sites 531:560 engineered glycosylation sites 531:560 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 9 41 theme evolution 1334:1342 arg1 rounds 1315:1320 Further rounds 1307:1320 Further rounds of in vitro evolution 1307:1342 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 7 42 theme homology 891:898 arg1 model 900:904 A three-dimensional homology model 871:904 A three-dimensional homology model 871:904 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 43 theme active 1127:1132 arg1 residue 1139:1145 an active site residue 1124:1145 an active site residue 1124:1145 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 6 44 theme PglB 865:868 arg1 engineering 850:860 structure-guided engineering 833:860 structure-guided engineering of PglB 833:868 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 7 45 theme potential 1033:1041 arg1 residues 1061:1068 potential sugar-interacting residues 1033:1068 potential sugar-interacting residues 1033:1068 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 7 45 theme potential 1033:1041 arg1 targets 1073:1079 targets 1073:1079 targets for mutagenesis 1073:1095 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 2 46 from expression 237:246 arg1 coli 331:334 Escherichia coli 319:334 Escherichia coli 319:334 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 8 47 theme residue 1139:1145 arg1 mutagenesis 1109:1119 Saturation mutagenesis 1098:1119 Saturation mutagenesis of an active site residue 1098:1145 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 9 48 theme triple 1353:1358 arg1 S80R-Q287P-N311V 1367:1382 a triple mutant S80R-Q287P-N311V 1351:1382 a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16 1351:1464 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 8 49 dep facilitated 1191:1201 arg1 increased 1223:1231 increased 1223:1231 facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA 1191:1304 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 2 50 theme simplified 346:355 arg1 production 357:366 a simplified production 344:366 a simplified production of glycoconjugate vaccines in prokaryotic cells 344:414 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 8 51 theme mutation 1169:1176 arg1 N311V 1178:1182 the enhancing mutation N311V 1155:1182 the enhancing mutation N311V 1155:1182 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 6 52 theme O 744:744 arg1 antigen 746:752 Typhimurium LT2 O antigen 728:752 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) 728:782 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 2 53 theme prokaryotic 398:408 arg1 cells 410:414 prokaryotic cells 398:414 prokaryotic cells 398:414 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 6 54 theme Typhimurium 728:738 arg1 LT2 740:742 Typhimurium LT2 728:742 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) 728:782 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 2 55 theme vaccines 386:393 arg1 production 357:366 a simplified production 344:366 a simplified production of glycoconjugate vaccines in prokaryotic cells 344:414 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 9 56 theme S. 1416:1417 arg1 LT2 1428:1430 S. enterica LT2 1416:1430 S. enterica LT2 glycoconjugates 1416:1446 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 2 57 theme jejuni 309:314 arg1 PglB 287:290 N-oligosaccharyltransferase (N-OST) PglB 251:290 N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni 251:314 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 4 58 theme PglBCj 586:591 arg1 efficiency 572:581 Transfer efficiency 563:581 Transfer efficiency of PglBCj 563:591 Transfer efficiency of PglBCj is low for certain heterologous polysaccharide substrates. 25833378 9 59 theme LT2 1428:1430 arg1 glycoconjugates 1432:1446 S. enterica LT2 glycoconjugates 1416:1446 S. enterica LT2 glycoconjugates 1416:1446 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 1 60 theme most 144:147 arg1 measures 170:177 the most efficient preventive measures 140:177 the most efficient preventive measures against life-threatening bacterial infections 140:223 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 1 61 theme preventive 159:168 arg1 measures 170:177 the most efficient preventive measures 140:177 the most efficient preventive measures against life-threatening bacterial infections 140:223 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 2 62 from production 357:366 arg1 cells 410:414 prokaryotic cells 398:414 prokaryotic cells 398:414 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 4 63 theme polysaccharide 625:638 arg1 substrates 640:649 certain heterologous polysaccharide substrates 604:649 certain heterologous polysaccharide substrates 604:649 Transfer efficiency of PglBCj is low for certain heterologous polysaccharide substrates. 25833378 9 64 theme 16 1463:1464 arg1 factor 1453:1458 a factor 1451:1458 a factor of 16 1451:1464 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 1 65 theme bacterial 204:212 arg1 infections 214:223 life-threatening bacterial infections 187:223 life-threatening bacterial infections 187:223 Conjugate vaccines belong to the most efficient preventive measures against life-threatening bacterial infections. 25833378 9 66 dep in 1325:1326 arg1 vitro 1328:1332 vitro 1328:1332 vitro 1328:1332 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 3 67 theme pathogenic 444:453 arg1 bacteria 455:462 pathogenic bacteria 444:462 pathogenic bacteria 444:462 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 4 68 theme certain 604:610 arg1 substrates 640:649 certain heterologous polysaccharide substrates 604:649 certain heterologous polysaccharide substrates 604:649 Transfer efficiency of PglBCj is low for certain heterologous polysaccharide substrates. 25833378 6 69 theme structure-guided 833:848 arg1 engineering 850:860 structure-guided engineering 833:860 structure-guided engineering of PglB 833:868 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 5 70 theme Salmonella 704:713 arg1 sv 724:725 Salmonella enterica sv 704:725 Salmonella enterica sv 704:725 In this study, we increased glycosylation rates for Salmonella enterica sv. 25833378 10 71 theme bacterial 1496:1504 arg1 N-OST 1506:1510 bacterial N-OST 1496:1510 bacterial N-OST 1496:1510 Our results demonstrate that bacterial N-OST can be tailored to specific polysaccharide substrates by structure-guided protein engineering. 25833378 3 72 theme glycosylation 542:554 arg1 sites 556:560 engineered glycosylation sites 531:560 engineered glycosylation sites 531:560 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 0 73 theme Campylobacter 24:36 arg1 jejuni 38:43 Campylobacter jejuni 24:43 Campylobacter jejuni 24:43 Increased efficiency of Campylobacter jejuni N-oligosaccharyltransferase PglB by structure-guided engineering. 25833378 7 74 theme jejuni 962:967 arg1 N-glycan 969:976 the natural C. jejuni N-glycan 947:976 the natural C. jejuni N-glycan attached to the acceptor peptide 947:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 10 75 theme polysaccharide 1540:1553 arg1 substrates 1555:1564 specific polysaccharide substrates 1531:1564 specific polysaccharide substrates 1531:1564 Our results demonstrate that bacterial N-OST can be tailored to specific polysaccharide substrates by structure-guided protein engineering. 25833378 6 76 theme aureus 803:808 arg1 CP5 810:812 Staphylococcus aureus CP5 788:812 Staphylococcus aureus CP5 788:812 Typhimurium LT2 O antigen (which lacks N-acetyl sugars) and Staphylococcus aureus CP5 polysaccharides by structure-guided engineering of PglB. 25833378 7 77 theme natural 951:957 arg1 jejuni 962:967 natural C. jejuni 951:967 the natural C. jejuni N-glycan attached to the acceptor peptide 947:1009 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 2 78 theme Functional 226:235 arg1 expression 237:246 Functional expression 226:246 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli 226:334 Functional expression of N-oligosaccharyltransferase (N-OST) PglB of Campylobacter jejuni in Escherichia coli enables a simplified production of glycoconjugate vaccines in prokaryotic cells. 25833378 9 79 theme Further 1307:1313 arg1 rounds 1315:1320 Further rounds 1307:1320 Further rounds of in vitro evolution 1307:1342 Further rounds of in vitro evolution led to a triple mutant S80R-Q287P-N311V enabling a yield improvement of S. enterica LT2 glycoconjugates by a factor of 16. 25833378 8 80 theme glycoprotein-specific 1278:1298 arg1 ELISA 1300:1304 glycoprotein-specific ELISA 1278:1304 glycoprotein-specific ELISA 1278:1304 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 3 81 theme acceptor 505:512 arg1 proteins 514:521 immunogenic acceptor proteins 493:521 immunogenic acceptor proteins bearing engineered glycosylation sites 493:560 Polysaccharide antigens of pathogenic bacteria can be covalently coupled to immunogenic acceptor proteins bearing engineered glycosylation sites. 25833378 7 82 theme sugar-interacting 1043:1059 arg1 residues 1061:1068 potential sugar-interacting residues 1033:1068 potential sugar-interacting residues 1033:1068 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 7 82 theme sugar-interacting 1043:1059 arg1 targets 1073:1079 targets 1073:1079 targets for mutagenesis 1073:1095 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 7 83 theme membrane-associated 909:927 arg1 PglBCj 929:934 membrane-associated PglBCj 909:934 membrane-associated PglBCj 909:934 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 7 84 used used 1016:1019 arg2 model 900:904 A three-dimensional homology model 871:904 A three-dimensional homology model 871:904 A three-dimensional homology model of membrane-associated PglBCj, docked to the natural C. jejuni N-glycan attached to the acceptor peptide, was used to identify potential sugar-interacting residues as targets for mutagenesis. 25833378 8 85 dep in 1233:1234 arg1 vivo 1236:1239 vivo 1236:1239 vivo 1236:1239 Saturation mutagenesis of an active site residue yielded the enhancing mutation N311V, which facilitated fivefold to 11-fold increased in vivo glycosylation rates as determined by glycoprotein-specific ELISA. 25833378 10 86 theme specific 1531:1538 arg1 substrates 1555:1564 specific polysaccharide substrates 1531:1564 specific polysaccharide substrates 1531:1564 Our results demonstrate that bacterial N-OST can be tailored to specific polysaccharide substrates by structure-guided protein engineering. 24889612 5 0 theme sTn 724:726 arg1 O-glycan 728:735 the sTn O-glycan 720:735 the sTn O-glycan 720:735 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 1 1 theme type 143:146 arg1 receptor 150:157 Paired Ig-like type 2 receptor α 128:159 Paired Ig-like type 2 receptor α (PILRα) 128:167 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 1 theme type 143:146 arg1 PILRα 162:166 PILRα 162:166 PILRα 162:166 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 7 2 theme mucin 1028:1032 arg1 proteins 1034:1041 O-glycosylated mucin proteins 1013:1041 O-glycosylated mucin proteins with sTn modifications 1013:1064 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 0 3 theme attached 69:76 arg1 peptide 78:84 its attached peptide 65:84 its attached peptide of mucin family 65:100 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 2 4 theme recognitions 322:333 arg1 characteristics 297:311 the molecular characteristics 283:311 the molecular characteristics of these recognitions 283:333 However, the molecular characteristics of these recognitions are largely unknown. 24889612 2 4 theme recognitions 322:333 arg1 unknown 347:353 unknown 347:353 unknown 347:353 However, the molecular characteristics of these recognitions are largely unknown. 24889612 1 5 theme related 221:227 arg1 proteins 229:236 related proteins 221:236 related proteins 221:236 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 0 6 theme mucin 89:93 arg1 family 95:100 mucin family 89:100 mucin family 89:100 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 3 7 gly sialylated 374:383 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 7 gly sialylated 374:383 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 1 8 theme proteins 229:236 arg1 range 187:191 a wide range 180:191 a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses 180:271 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 9 gly O-glycosylated 196:209 arg1 mucin 211:215 O-glycosylated mucin 196:215 O-glycosylated mucin 196:215 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 4 10 theme peptide 603:609 arg1 region 611:616 its attached peptide region 590:616 its attached peptide region 590:616 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 0 11 theme family 95:100 arg1 peptide 78:84 its attached peptide 65:84 its attached peptide of mucin family 65:100 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 0 11 theme family 95:100 arg1 recognition 34:44 simultaneous recognition 21:44 simultaneous recognition of an O-glycan 21:59 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 7 12 theme immune 1088:1093 arg1 system 1095:1100 the immune system 1084:1100 the immune system 1084:1100 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 13 theme binding 913:919 arg1 distinct 961:968 distinct 961:968 distinct 961:968 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 13 theme binding 913:919 arg1 motif 921:925 the binding motif 909:925 the binding motif 909:925 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 4 14 theme attached 594:601 arg1 region 611:616 its attached peptide region 590:616 its attached peptide region 590:616 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 6 15 theme binding 847:853 arg1 mode 855:858 this binding mode 842:858 this binding mode 842:858 Binding and functional assays support this binding mode. 24889612 3 16 dep required 457:464 arg1 both 452:455 both 452:455 both 452:455 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 17 theme T 400:400 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 17 theme T 400:400 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 0 18 theme Structural 0:9 arg1 basis 11:15 Structural basis 0:15 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 0:126 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 7 19 theme sTn 1048:1050 arg1 modifications 1052:1064 sTn modifications 1048:1064 sTn modifications 1048:1064 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 20 with proteins 1034:1041 arg1 modifications 1052:1064 sTn modifications 1048:1064 sTn modifications 1048:1064 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 0 21 theme simultaneous 21:32 arg1 recognition 34:44 simultaneous recognition 21:44 simultaneous recognition of an O-glycan 21:59 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 1 22 theme broad 250:254 arg1 responses 263:271 broad immune responses 250:271 broad immune responses 250:271 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 0 23 theme receptor 112:119 arg1 PILRα 121:125 immune receptor PILRα 105:125 immune receptor PILRα 105:125 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 7 24 theme molecular 931:939 arg1 mechanism 941:949 molecular mechanism 931:949 molecular mechanism 931:949 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 5 25 theme compact 745:751 arg1 structure 761:769 the compact peptide structure 741:769 the compact peptide structure constrained by proline residues 741:801 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 5 26 theme proline 786:792 arg1 residues 794:801 proline residues 786:801 proline residues 786:801 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 1 27 theme immune 256:261 arg1 responses 263:271 broad immune responses 250:271 broad immune responses 250:271 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 0 28 theme immune 105:110 arg1 PILRα 121:125 immune receptor PILRα 105:125 immune receptor PILRα 105:125 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 3 29 theme ligand 470:475 arg1 recognition 477:487 ligand recognition 470:487 ligand recognition by PILRα 470:496 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 30 theme O-linked 385:392 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 30 theme O-linked 385:392 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 5 31 theme large 659:663 arg1 change 680:685 large conformational change 659:685 large conformational change 659:685 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 5 32 theme conformational 665:678 arg1 change 680:685 large conformational change 659:685 large conformational change 659:685 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 7 33 from receptors 993:1001 arg1 distinct 961:968 distinct 961:968 distinct 961:968 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 33 from receptors 993:1001 arg1 motif 921:925 the binding motif 909:925 the binding motif 909:925 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 3 34 theme sugar 394:398 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 34 theme sugar 394:398 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 4 35 theme crystal 530:536 arg1 structures 538:547 the crystal structures 526:547 the crystal structures of PILRα 526:556 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 4 36 with complex 566:572 arg1 region 611:616 its attached peptide region 590:616 its attached peptide region 590:616 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 4 36 with complex 566:572 arg1 sTn 582:584 an sTn 579:584 an sTn 579:584 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 1 37 theme wide 182:185 arg1 range 187:191 a wide range 180:191 a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses 180:271 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 0 38 theme O-glycan 52:59 arg1 peptide 78:84 its attached peptide 65:84 its attached peptide of mucin family 65:100 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 0 38 theme O-glycan 52:59 arg1 recognition 34:44 simultaneous recognition 21:44 simultaneous recognition of an O-glycan 21:59 Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. 24889612 5 39 theme peptide 753:759 arg1 structure 761:769 the compact peptide structure 741:769 the compact peptide structure constrained by proline residues 741:801 The structures show that PILRα exhibits large conformational change to recognize simultaneously both the sTn O-glycan and the compact peptide structure constrained by proline residues. 24889612 6 40 theme functional 816:825 arg1 assays 827:832 functional assays 816:832 functional assays 816:832 Binding and functional assays support this binding mode. 24889612 3 41 link O-linked 385:392 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 41 link O-linked 385:392 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 42 theme attached 424:431 arg1 region 441:446 its attached peptide region 420:446 its attached peptide region 420:446 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 43 theme sialylated 374:383 arg1 sTn 411:413 sTn 411:413 sTn 411:413 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 3 43 theme sialylated 374:383 arg1 antigen 402:408 O-linked sugar T antigen 385:408 sialylated O-linked sugar T antigen (sTn) 374:414 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 7 44 theme sugar-recognition 975:991 arg1 receptors 993:1001 sugar-recognition receptors 975:1001 sugar-recognition receptors 975:1001 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 45 theme significant 884:894 arg1 insight 896:902 significant insight 884:902 significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry 884:1130 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 7 46 theme viral 1120:1124 arg1 entry 1126:1130 viral entry 1120:1130 viral entry 1120:1130 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 3 47 theme peptide 433:439 arg1 region 441:446 its attached peptide region 420:446 its attached peptide region 420:446 Here we show that sialylated O-linked sugar T antigen (sTn) and its attached peptide region are both required for ligand recognition by PILRα. 24889612 7 48 theme O-glycosylated 1013:1026 arg1 proteins 1034:1041 O-glycosylated mucin proteins 1013:1041 O-glycosylated mucin proteins with sTn modifications 1013:1064 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 2 49 theme molecular 287:295 arg1 characteristics 297:311 the molecular characteristics 283:311 the molecular characteristics of these recognitions 283:333 However, the molecular characteristics of these recognitions are largely unknown. 24889612 2 49 theme molecular 287:295 arg1 unknown 347:353 unknown 347:353 unknown 347:353 However, the molecular characteristics of these recognitions are largely unknown. 24889612 7 50 gly O-glycosylated 1013:1026 arg1 proteins 1034:1041 O-glycosylated mucin proteins 1013:1041 O-glycosylated mucin proteins with sTn modifications 1013:1064 These findings provide significant insight into the binding motif and molecular mechanism (which is distinct from sugar-recognition receptors) by which O-glycosylated mucin proteins with sTn modifications are recognized in the immune system as well as during viral entry. 24889612 4 51 with structures 538:547 arg1 region 611:616 its attached peptide region 590:616 its attached peptide region 590:616 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 4 51 with structures 538:547 arg1 sTn 582:584 an sTn 579:584 an sTn 579:584 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 1 52 theme Paired 128:133 arg1 receptor 150:157 Paired Ig-like type 2 receptor α 128:159 Paired Ig-like type 2 receptor α (PILRα) 128:167 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 52 theme Paired 128:133 arg1 PILRα 162:166 PILRα 162:166 PILRα 162:166 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 53 theme O-glycosylated 196:209 arg1 mucin 211:215 O-glycosylated mucin 196:215 O-glycosylated mucin 196:215 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 4 54 theme PILRα 552:556 arg1 complex 566:572 its complex 562:572 its complex with an sTn and its attached peptide region 562:616 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 4 54 theme PILRα 552:556 arg1 structures 538:547 the crystal structures 526:547 the crystal structures of PILRα 526:556 Furthermore, we determined the crystal structures of PILRα and its complex with an sTn and its attached peptide region. 24889612 1 55 theme Ig-like 135:141 arg1 receptor 150:157 Paired Ig-like type 2 receptor α 128:159 Paired Ig-like type 2 receptor α (PILRα) 128:167 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 55 theme Ig-like 135:141 arg1 PILRα 162:166 PILRα 162:166 PILRα 162:166 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 24889612 1 56 theme mucin 211:215 arg1 range 187:191 a wide range 180:191 a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses 180:271 Paired Ig-like type 2 receptor α (PILRα) recognizes a wide range of O-glycosylated mucin and related proteins to regulate broad immune responses. 25655580 9 0 theme discovery 1267:1275 arg1 datasets 1277:1284 discovery datasets 1267:1284 discovery datasets 1267:1284 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 6 1 theme related 813:819 arg1 expression 802:811 altered gene expression 789:811 altered gene expression related to glycosaminoglycans (GAGs) and N-glycans 789:862 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 3 2 theme carcinomas 494:503 arg1 datasets 462:469 Five expression datasets 446:469 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples 446:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 8 3 theme Subtype-specific 1036:1051 arg1 patterns 1064:1071 Subtype-specific expression patterns 1036:1071 Subtype-specific expression patterns 1036:1071 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 4 4 theme breast 632:637 arg1 carcinomas 639:648 1960 breast carcinomas 627:648 1960 breast carcinomas 627:648 Results were validated in 1960 breast carcinomas. 25655580 10 5 theme UGCG 1530:1533 arg1 association 1504:1514 association 1504:1514 association of B3GNT5 and UGCG genes to patient survival 1504:1559 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 3 6 theme in 527:528 arg1 carcinomas 516:525 31 ductal carcinomas 506:525 31 ductal carcinomas in situ (DCIS) 506:540 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 3 6 theme in 527:528 arg1 DCIS 536:539 DCIS 536:539 DCIS 536:539 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 6 7 theme non-malignant 881:893 arg1 samples 895:901 non-malignant samples 881:901 non-malignant samples 881:901 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 10 8 theme B3GNT5 1519:1524 arg1 association 1504:1514 association 1504:1514 association of B3GNT5 and UGCG genes to patient survival 1504:1559 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 13 9 theme cancer 1918:1923 arg1 patients 1925:1932 breast cancer patients 1911:1932 breast cancer patients 1911:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 12 10 theme prognostic 1740:1749 arg1 signature 1751:1759 a prognostic signature 1738:1759 a prognostic signature for two basal-like subgroups 1738:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 13 11 theme prognostication 1863:1877 arg1 stratification 1893:1906 better prognostication and treatment stratification 1856:1906 better prognostication and treatment stratification of breast cancer patients 1856:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 3 12 theme tissue 571:576 arg1 samples 578:584 79 non-malignant breast tissue samples 547:584 79 non-malignant breast tissue samples 547:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 13 13 theme treatment 1883:1891 arg1 stratification 1893:1906 better prognostication and treatment stratification 1856:1906 better prognostication and treatment stratification of breast cancer patients 1856:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 3 14 theme non-malignant 550:562 arg1 samples 578:584 79 non-malignant breast tissue samples 547:584 79 non-malignant breast tissue samples 547:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 3 15 dep in 527:528 arg1 situ 530:533 situ 530:533 situ 530:533 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 12 16 theme cancer 1712:1717 arg1 subtypes 1719:1726 breast cancer subtypes 1705:1726 breast cancer subtypes including a prognostic signature for two basal-like subgroups 1705:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 12 16 theme cancer 1712:1717 arg1 signature 1751:1759 a prognostic signature 1738:1759 a prognostic signature for two basal-like subgroups 1738:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 7 17 from changes 974:980 arg1 sulfation 1003:1011 glycosaminoglycan sulfation 985:1011 glycosaminoglycan sulfation 985:1011 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 7 17 from changes 974:980 arg1 proteins 1026:1033 adhesion proteins 1017:1033 adhesion proteins 1017:1033 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 5 18 theme glycosylation-related 670:690 arg1 proteins 692:699 glycosylation-related proteins 670:699 glycosylation-related proteins 670:699 419 genes encoding glycosylation-related proteins were selected. 25655580 2 19 from differences 274:284 arg1 expression 289:298 expression 289:298 expression of genes related to the process of glycosylation 289:347 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 11 20 theme glycan-specific 1582:1596 arg1 changes 1598:1604 most glycan-specific changes 1577:1604 most glycan-specific changes 1577:1604 In conclusion, most glycan-specific changes occur early in the carcinogenic process. 25655580 1 21 theme malignancy 143:152 arg1 Alterations 91:101 Alterations 91:101 Alterations in glycan structures 91:122 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 1 21 theme malignancy 143:152 arg1 signs 134:138 early signs 128:138 early signs of malignancy 128:152 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 1 22 from force 210:214 arg1 part 195:198 part 195:198 part 195:198 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 11 23 theme carcinogenic 1625:1636 arg1 process 1638:1644 the carcinogenic process 1621:1644 the carcinogenic process 1621:1644 In conclusion, most glycan-specific changes occur early in the carcinogenic process. 25655580 8 24 theme genes 1101:1105 arg1 down-regulation 1082:1096 down-regulation 1082:1096 down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes 1082:1170 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 8 25 theme glycan-binding 1116:1129 arg1 proteins 1131:1138 glycan-binding proteins 1116:1138 glycan-binding proteins 1116:1138 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 7 26 theme glycosaminoglycan 985:1001 arg1 sulfation 1003:1011 glycosaminoglycan sulfation 985:1011 glycosaminoglycan sulfation 985:1011 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 2 27 theme carcinoma 370:378 arg1 subtypes 380:387 breast carcinoma subtypes 363:387 breast carcinoma subtypes 363:387 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 0 28 theme Glycan-related 0:13 arg1 signatures 31:40 Glycan-related gene expression signatures 0:40 Glycan-related gene expression signatures in breast cancer subtypes 0:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 8 29 theme luminal 1147:1153 arg1 subtypes 1163:1170 the luminal A and B subtypes 1143:1170 the luminal A and B subtypes 1143:1170 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 9 30 theme basal-like 1184:1193 arg1 samples 1195:1201 Clustering basal-like samples 1173:1201 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets 1173:1284 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 3 31 theme expression 451:460 arg1 datasets 462:469 Five expression datasets 446:469 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples 446:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 0 32 theme expression 20:29 arg1 signatures 31:40 Glycan-related gene expression signatures 0:40 Glycan-related gene expression signatures in breast cancer subtypes 0:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 1 33 theme glycan 106:111 arg1 structures 113:122 glycan structures 106:122 glycan structures 106:122 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 9 34 theme consensus 1211:1219 arg1 list 1221:1224 a consensus list 1209:1224 a consensus list of genes differentially expressed across discovery datasets 1209:1284 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 3 35 theme invasive 478:485 arg1 carcinomas 494:503 454 invasive breast carcinomas 474:503 454 invasive breast carcinomas 474:503 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 0 36 theme cancer 52:57 arg1 subtypes 59:66 breast cancer subtypes 45:66 breast cancer subtypes 45:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 6 37 theme gene 797:800 arg1 expression 802:811 altered gene expression 789:811 altered gene expression related to glycosaminoglycans (GAGs) and N-glycans 789:862 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 10 38 theme important 1433:1441 arg1 role 1443:1446 an important role 1430:1446 an important role 1430:1446 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 7 39 theme aggressiveness 935:948 arg1 potentials 950:959 different aggressiveness potentials 925:959 different aggressiveness potentials 925:959 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 1 40 theme driving 202:208 arg1 Alterations 91:101 Alterations 91:101 Alterations in glycan structures 91:122 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 1 40 theme driving 202:208 arg1 force 210:214 a driving force 200:214 a driving force behind malignant transformation 200:246 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 10 41 theme tumors 1476:1481 arg1 carcinogenesis 1451:1464 carcinogenesis 1451:1464 carcinogenesis of breast tumors 1451:1481 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 1 42 from Alterations 91:101 arg1 structures 113:122 glycan structures 106:122 glycan structures 106:122 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 8 43 from down-regulation 1082:1096 arg1 subtypes 1163:1170 the luminal A and B subtypes 1143:1170 the luminal A and B subtypes 1143:1170 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 6 44 dep similar 758:764 arg1 showing 781:787 showing 781:787 showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans 781:862 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 2 45 theme glycosylation 335:347 arg1 process 324:330 the process 320:330 the process of glycosylation 320:347 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 7 46 theme In-situ 904:910 arg1 lesions 912:918 In-situ lesions 904:918 In-situ lesions with different aggressiveness potentials 904:959 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 13 47 from research 1798:1805 arg1 area 1815:1818 this area 1810:1818 this area 1810:1818 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 13 48 theme patients 1925:1932 arg1 stratification 1893:1906 better prognostication and treatment stratification 1856:1906 better prognostication and treatment stratification of breast cancer patients 1856:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 7 49 with lesions 912:918 arg1 potentials 950:959 different aggressiveness potentials 925:959 different aggressiveness potentials 925:959 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 8 50 theme expression 1053:1062 arg1 patterns 1064:1071 Subtype-specific expression patterns 1036:1071 Subtype-specific expression patterns 1036:1071 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 3 51 theme carcinomas 516:525 arg1 datasets 462:469 Five expression datasets 446:469 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples 446:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 6 52 theme altered 789:795 arg1 expression 802:811 altered gene expression 789:811 altered gene expression related to glycosaminoglycans (GAGs) and N-glycans 789:862 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 2 53 theme genes 303:307 arg1 expression 289:298 expression 289:298 expression of genes related to the process of glycosylation 289:347 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 13 54 theme breast 1911:1916 arg1 patients 1925:1932 breast cancer patients 1911:1932 breast cancer patients 1911:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 13 55 theme better 1856:1861 arg1 stratification 1893:1906 better prognostication and treatment stratification 1856:1906 better prognostication and treatment stratification of breast cancer patients 1856:1932 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 3 56 theme breast 564:569 arg1 samples 578:584 79 non-malignant breast tissue samples 547:584 79 non-malignant breast tissue samples 547:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 7 57 theme different 925:933 arg1 potentials 950:959 different aggressiveness potentials 925:959 different aggressiveness potentials 925:959 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 12 58 theme breast 1705:1710 arg1 subtypes 1719:1726 breast cancer subtypes 1705:1726 breast cancer subtypes including a prognostic signature for two basal-like subgroups 1705:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 12 58 theme breast 1705:1710 arg1 signature 1751:1759 a prognostic signature 1738:1759 a prognostic signature for two basal-like subgroups 1738:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 9 59 theme validation 1354:1363 arg1 dataset 1365:1371 the validation dataset 1350:1371 the validation dataset 1350:1371 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 1 60 theme early 128:132 arg1 Alterations 91:101 Alterations 91:101 Alterations in glycan structures 91:122 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 1 60 theme early 128:132 arg1 signs 134:138 early signs 128:138 early signs of malignancy 128:152 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 25655580 12 61 theme glycan-related 1666:1679 arg1 alterations 1681:1691 glycan-related alterations 1666:1691 glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups 1666:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 11 62 theme most 1577:1580 arg1 changes 1598:1604 most glycan-specific changes 1577:1604 most glycan-specific changes 1577:1604 In conclusion, most glycan-specific changes occur early in the carcinogenic process. 25655580 3 63 theme samples 578:584 arg1 datasets 462:469 Five expression datasets 446:469 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples 446:584 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 12 64 theme specific 1693:1700 arg1 alterations 1681:1691 glycan-related alterations 1666:1691 glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups 1666:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 0 65 from signatures 31:40 arg1 subtypes 59:66 breast cancer subtypes 45:66 breast cancer subtypes 45:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 10 66 theme patient 1544:1550 arg1 survival 1552:1559 patient survival 1544:1559 patient survival 1544:1559 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 7 67 theme adhesion 1017:1024 arg1 proteins 1026:1033 adhesion proteins 1017:1033 adhesion proteins 1017:1033 In-situ lesions with different aggressiveness potentials demonstrated changes in glycosaminoglycan sulfation and adhesion proteins. 25655580 0 68 theme gene 15:18 arg1 signatures 31:40 Glycan-related gene expression signatures 0:40 Glycan-related gene expression signatures in breast cancer subtypes 0:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 8 69 theme A 1155:1155 arg1 subtypes 1163:1170 the luminal A and B subtypes 1143:1170 the luminal A and B subtypes 1143:1170 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 2 70 theme clinical 425:432 arg1 parameters 434:443 clinical parameters 425:443 clinical parameters 425:443 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 9 71 theme Clustering 1173:1182 arg1 samples 1195:1201 Clustering basal-like samples 1173:1201 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets 1173:1284 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 2 72 theme breast 363:368 arg1 subtypes 380:387 breast carcinoma subtypes 363:387 breast carcinoma subtypes 363:387 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 8 73 theme B 1161:1161 arg1 subtypes 1163:1170 the luminal A and B subtypes 1143:1170 the luminal A and B subtypes 1143:1170 Subtype-specific expression patterns revealed down-regulation of genes encoding glycan-binding proteins in the luminal A and B subtypes. 25655580 0 74 dep relation 69:76 arg1 signatures 31:40 Glycan-related gene expression signatures 0:40 Glycan-related gene expression signatures in breast cancer subtypes 0:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 13 75 theme Future 1791:1796 arg1 research 1798:1805 Future research 1791:1805 Future research in this area 1791:1818 Future research in this area may potentially lead to markers for better prognostication and treatment stratification of breast cancer patients. 25655580 0 76 theme breast 45:50 arg1 subtypes 59:66 breast cancer subtypes 45:66 breast cancer subtypes 45:66 Glycan-related gene expression signatures in breast cancer subtypes; relation to survival. 25655580 12 77 theme basal-like 1769:1778 arg1 subgroups 1780:1788 two basal-like subgroups 1765:1788 two basal-like subgroups 1765:1788 We have identified glycan-related alterations specific to breast cancer subtypes including a prognostic signature for two basal-like subgroups. 25655580 3 78 theme breast 487:492 arg1 carcinomas 494:503 454 invasive breast carcinomas 474:503 454 invasive breast carcinomas 474:503 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 10 79 dep B3GNT5 1519:1524 arg1 genes 1535:1539 genes 1535:1539 genes 1535:1539 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 6 80 theme DCIS 720:723 arg1 samples 725:731 The DCIS samples 716:731 The DCIS samples 716:731 The DCIS samples appeared expression-wise similar to carcinomas, showing altered gene expression related to glycosaminoglycans (GAGs) and N-glycans when compared to non-malignant samples. 25655580 9 81 theme genes 1229:1233 arg1 list 1221:1224 a consensus list 1209:1224 a consensus list of genes differentially expressed across discovery datasets 1209:1284 Clustering basal-like samples using a consensus list of genes differentially expressed across discovery datasets produced two clusters with significantly differing prognosis in the validation dataset. 25655580 2 82 theme related 309:315 arg1 genes 303:307 genes 303:307 genes related to the process of glycosylation 303:347 Here, we explore whether differences in expression of genes related to the process of glycosylation exist between breast carcinoma subtypes - and look for their association to clinical parameters. 25655580 10 83 theme breast 1469:1474 arg1 tumors 1476:1481 breast tumors 1469:1481 breast tumors 1469:1481 Finally, our analyses suggest that glycolipids may play an important role in carcinogenesis of breast tumors - as demonstrated by association of B3GNT5 and UGCG genes to patient survival. 25655580 3 84 theme ductal 509:514 arg1 carcinomas 516:525 31 ductal carcinomas 506:525 31 ductal carcinomas in situ (DCIS) 506:540 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 3 84 theme ductal 509:514 arg1 DCIS 536:539 DCIS 536:539 DCIS 536:539 Five expression datasets of 454 invasive breast carcinomas, 31 ductal carcinomas in situ (DCIS), and 79 non-malignant breast tissue samples were analysed. 25655580 1 85 theme malignant 223:231 arg1 transformation 233:246 malignant transformation 223:246 malignant transformation 223:246 Alterations in glycan structures are early signs of malignancy and have recently been proposed to be in part a driving force behind malignant transformation. 28800704 3 0 from research 696:703 arg1 role 712:715 the role 708:715 the role played by N-glycans in Ginkgo-caused allergy 708:760 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 7 1 theme types 1545:1549 arg1 amount 1489:1494 the absolute amount 1476:1494 the absolute amount of each N-glycan 1476:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 1 theme types 1545:1549 arg1 N-glycan 1504:1511 each N-glycan 1499:1511 each N-glycan 1499:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 1 theme types 1545:1549 arg1 percentage 1521:1530 the percentage 1517:1530 the percentage of different types of N-glycan 1517:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 1 theme types 1545:1549 arg1 types 1545:1549 different types 1535:1549 different types of N-glycan 1535:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 1 2 theme cross-reactive 170:183 arg1 CCDs 199:202 CCDs 199:202 CCDs 199:202 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 1 2 theme cross-reactive 170:183 arg1 determinants 185:196 carbohydrate cross-reactive determinants 157:196 carbohydrate cross-reactive determinants (CCDs) 157:203 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 5 3 theme ultraperformance 974:989 arg1 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 3 theme ultraperformance 974:989 arg1 chromatography 998:1011 hydrophilic interaction ultraperformance liquid chromatography 950:1011 hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) 950:1024 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 2 4 from allergen 354:361 arg1 seeds 380:384 Ginkgo biloba seeds 366:384 Ginkgo biloba seeds 366:384 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 2 5 theme biloba 373:378 arg1 seeds 380:384 Ginkgo biloba seeds 366:384 Ginkgo biloba seeds 366:384 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 7 6 theme geographic 1633:1642 arg1 variation 1644:1652 geographic variation 1633:1652 geographic variation 1633:1652 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 0 7 theme biloba 105:110 arg1 Seeds 89:93 Seeds 89:93 Seeds of Ginkgo biloba 89:110 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 7 8 theme significant 1570:1580 arg1 diversity 1582:1590 significant diversity 1570:1590 significant diversity among the samples without any pattern of geographic variation 1570:1652 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 6 9 theme complex-type 1247:1258 arg1 N-glycans 1260:1268 complex-type N-glycans 1247:1268 complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans 1247:1329 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 5 10 theme matrix-assisted 1030:1044 arg1 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 10 theme matrix-assisted 1030:1044 arg1 spectrometry 1094:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1030:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 1030:1120 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 1 11 theme innate 285:290 arg1 system 299:304 the innate immune system 281:304 the innate immune system 281:304 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 0 12 theme Ginkgo 98:103 arg1 biloba 105:110 Ginkgo biloba 98:110 Ginkgo biloba 98:110 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 3 13 theme Ginkgo-caused 740:752 arg1 allergy 754:760 Ginkgo-caused allergy 740:760 Ginkgo-caused allergy 740:760 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 14 theme N-linked 566:573 arg1 moieties 545:552 carbohydrate moieties 532:552 carbohydrate moieties 532:552 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 14 theme N-linked 566:573 arg1 glycans 575:581 N-linked glycans 566:581 especially N-linked glycans 555:581 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 6 15 theme first 1373:1377 arg1 time 1379:1382 the first time 1369:1382 the first time 1369:1382 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 2 16 theme Previous 307:314 arg1 research 316:323 Previous research 307:323 Previous research 307:323 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 7 17 theme standard 1457:1464 arg1 use 1438:1440 use 1438:1440 use of an external standard 1438:1464 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 1 18 theme immune 292:297 arg1 system 299:304 the innate immune system 281:304 the innate immune system 281:304 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 3 19 gly glycoproteins 587:599 arg1 glycoproteins 587:599 glycoproteins 587:599 glycoproteins from Ginkgo seeds originating from different places for detailed structures 587:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 4 20 theme assays 820:825 arg1 Results 763:769 Results 763:769 Results of monosaccharide composition and immunoblotting assays 763:825 Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. 28800704 6 21 theme tandem 1209:1214 arg1 spectrometry 1221:1232 tandem mass spectrometry 1209:1232 tandem mass spectrometry 1209:1232 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 3 22 link N-linked 566:573 arg1 moieties 545:552 carbohydrate moieties 532:552 carbohydrate moieties 532:552 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 22 link N-linked 566:573 arg1 glycans 575:581 N-linked glycans 566:581 especially N-linked glycans 555:581 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 5 23 theme hydrophilic 950:960 arg1 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 23 theme hydrophilic 950:960 arg1 chromatography 998:1011 hydrophilic interaction ultraperformance liquid chromatography 950:1011 hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) 950:1024 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 4 24 theme immunoblotting 805:818 arg1 assays 820:825 immunoblotting assays 805:825 immunoblotting assays 805:825 Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. 28800704 3 25 theme carbohydrate 532:543 arg1 moieties 545:552 carbohydrate moieties 532:552 carbohydrate moieties 532:552 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 25 theme carbohydrate 532:543 arg1 glycans 575:581 N-linked glycans 566:581 especially N-linked glycans 555:581 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 7 26 theme N-glycan 1554:1561 arg1 types 1545:1549 different types 1535:1549 different types of N-glycan 1535:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 0 27 theme Qualitative 0:10 arg1 Analysis 29:36 Qualitative and Quantitative Analysis 0:36 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 0:111 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 6 28 theme Ginkgo 1352:1357 arg1 seeds 1359:1363 Ginkgo seeds 1352:1363 Ginkgo seeds for the first time 1352:1382 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 1 29 from progress 120:127 arg1 relationship 136:147 the relationship 132:147 the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response 132:225 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 3 30 theme glycoproteins 587:599 arg1 moieties 545:552 carbohydrate moieties 532:552 carbohydrate moieties 532:552 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 30 theme glycoproteins 587:599 arg1 glycans 575:581 N-linked glycans 566:581 especially N-linked glycans 555:581 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 7 31 theme variation 1644:1652 arg1 pattern 1622:1628 any pattern 1618:1628 any pattern of geographic variation 1618:1652 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 5 32 theme Detailed 865:872 arg1 elucidation 885:895 Detailed structural elucidation 865:895 Detailed structural elucidation of the N-glycans 865:912 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 7 33 theme N-glycan 1504:1511 arg1 amount 1489:1494 the absolute amount 1476:1494 the absolute amount of each N-glycan 1476:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 33 theme N-glycan 1504:1511 arg1 N-glycan 1504:1511 each N-glycan 1499:1511 each N-glycan 1499:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 33 theme N-glycan 1504:1511 arg1 percentage 1521:1530 the percentage 1517:1530 the percentage of different types of N-glycan 1517:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 33 theme N-glycan 1504:1511 arg1 types 1545:1549 different types 1535:1549 different types of N-glycan 1535:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 4 34 theme monosaccharide 774:787 arg1 composition 789:799 monosaccharide composition 774:799 monosaccharide composition 774:799 Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. 28800704 1 35 theme allergic 209:216 arg1 response 218:225 allergic response 209:225 allergic response 209:225 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 0 36 theme Quantitative 16:27 arg1 Analysis 29:36 Qualitative and Quantitative Analysis 0:36 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 0:111 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 2 37 gly glycosylated 389:400 arg1 allergen 354:361 the protein allergen 342:361 the protein allergen in Ginkgo biloba seeds 342:384 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 6 38 theme Lewis 1278:1282 arg1 determinants 1286:1297 Lewis A determinants 1278:1297 Lewis A determinants 1278:1297 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 1 39 theme Recent 113:118 arg1 progress 120:127 Recent progress 113:127 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response 113:225 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 3 40 from seeds 613:617 arg1 moieties 545:552 carbohydrate moieties 532:552 carbohydrate moieties 532:552 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 40 from seeds 613:617 arg1 glycans 575:581 N-linked glycans 566:581 especially N-linked glycans 555:581 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 3 40 from seeds 613:617 arg1 glycoproteins 587:599 glycoproteins 587:599 glycoproteins from Ginkgo seeds originating from different places for detailed structures 587:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 0 41 theme Carbohydrate 41:52 arg1 Modification 54:65 Carbohydrate Modification 41:65 Carbohydrate Modification 41:65 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 3 42 theme study 511:515 arg1 aim 499:501 The aim 495:501 The aim of this study 495:515 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 0 43 from Seeds 89:93 arg1 Glycoproteins 70:82 Glycoproteins 70:82 Glycoproteins from Seeds of Ginkgo biloba 70:110 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 7 44 theme Precise 1385:1391 arg1 quantification 1393:1406 Precise quantification 1385:1406 Precise quantification of N-glycans 1385:1419 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 3 45 theme detailed 657:664 arg1 structures 666:675 detailed structures 657:675 detailed structures 657:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 4 46 theme N-glycans 854:862 arg1 existence 841:849 the existence 837:849 the existence of N-glycans 837:862 Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. 28800704 5 47 theme N-glycans 904:912 arg1 elucidation 885:895 Detailed structural elucidation 865:895 Detailed structural elucidation of the N-glycans 865:912 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 48 theme structural 874:883 arg1 elucidation 885:895 Detailed structural elucidation 865:895 Detailed structural elucidation of the N-glycans 865:912 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 4 49 theme composition 789:799 arg1 Results 763:769 Results 763:769 Results of monosaccharide composition and immunoblotting assays 763:825 Results of monosaccharide composition and immunoblotting assays indicated the existence of N-glycans. 28800704 5 50 theme laser 1046:1050 arg1 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 50 theme laser 1046:1050 arg1 spectrometry 1094:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1030:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 1030:1120 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 7 51 theme different 1535:1543 arg1 types 1545:1549 different types 1535:1549 different types of N-glycan 1535:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 2 52 theme Ginkgo 366:371 arg1 seeds 380:384 Ginkgo biloba seeds 366:384 Ginkgo biloba seeds 366:384 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 0 53 theme Modification 54:65 arg1 Analysis 29:36 Qualitative and Quantitative Analysis 0:36 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 0:111 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 3 54 theme Ginkgo 606:611 arg1 seeds 613:617 Ginkgo seeds 606:617 Ginkgo seeds originating from different places for detailed structures 606:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 2 55 dep pointed 325:331 arg1 out 333:335 out 333:335 out 333:335 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 6 56 theme A 1284:1284 arg1 determinants 1286:1297 Lewis A determinants 1278:1297 Lewis A determinants 1278:1297 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 1 57 from importance 242:251 arg1 system 299:304 the innate immune system 281:304 the innate immune system 281:304 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 6 58 theme high-mannose-type 1303:1319 arg1 N-glycans 1321:1329 high-mannose-type N-glycans 1303:1329 high-mannose-type N-glycans 1303:1329 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 5 59 theme mass 1089:1092 arg1 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 59 theme mass 1089:1092 arg1 spectrometry 1094:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1030:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 1030:1120 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 7 60 theme absolute 1480:1487 arg1 amount 1489:1494 the absolute amount 1476:1494 the absolute amount of each N-glycan 1476:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 60 theme absolute 1480:1487 arg1 N-glycan 1504:1511 each N-glycan 1499:1511 each N-glycan 1499:1511 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 7 60 theme absolute 1480:1487 arg1 types 1545:1549 different types 1535:1549 different types of N-glycan 1535:1561 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 5 61 theme liquid 991:996 arg1 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 61 theme liquid 991:996 arg1 chromatography 998:1011 hydrophilic interaction ultraperformance liquid chromatography 950:1011 hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) 950:1024 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 62 theme time-of-flight 1074:1087 arg1 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 62 theme time-of-flight 1074:1087 arg1 spectrometry 1094:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1030:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 1030:1120 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 63 theme interaction 962:972 arg1 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 HILIC-UPLC 1014:1023 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 63 theme interaction 962:972 arg1 chromatography 998:1011 hydrophilic interaction ultraperformance liquid chromatography 950:1011 hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) 950:1024 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 64 theme desorption/ionization 1052:1072 arg1 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 MALDI-TOF-MS 1108:1119 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 5 64 theme desorption/ionization 1052:1072 arg1 spectrometry 1094:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1030:1105 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 1030:1120 Detailed structural elucidation of the N-glycans was further carried out by means of hydrophilic interaction ultraperformance liquid chromatography (HILIC-UPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 28800704 2 65 theme protein 346:352 arg1 allergen 354:361 the protein allergen 342:361 the protein allergen in Ginkgo biloba seeds 342:384 Previous research pointed out that the protein allergen in Ginkgo biloba seeds is glycosylated, and the oligosaccharides conjugated to these proteins might also contribute to the allergy. 28800704 6 66 theme mass 1216:1219 arg1 spectrometry 1221:1232 tandem mass spectrometry 1209:1232 tandem mass spectrometry 1209:1232 In total, 14 out of 16 structures detected by UPLC were confirmed by MALDI-TOF-MS and tandem mass spectrometry, among which complex-type N-glycans bearing Lewis A determinants and high-mannose-type N-glycans were identified from Ginkgo seeds for the first time. 28800704 3 67 theme further 688:694 arg1 research 696:703 further research 688:703 further research on the role played by N-glycans in Ginkgo-caused allergy 688:760 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 7 68 theme N-glycans 1411:1419 arg1 quantification 1393:1406 Precise quantification 1385:1406 Precise quantification of N-glycans 1385:1419 Precise quantification of N-glycans was performed by use of an external standard, and both the absolute amount of each N-glycan and the percentage of different types of N-glycan showed significant diversity among the samples without any pattern of geographic variation. 28800704 1 69 theme carbohydrate 256:267 arg1 moieties 269:276 carbohydrate moieties 256:276 carbohydrate moieties 256:276 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 0 70 from Analysis 29:36 arg1 Glycoproteins 70:82 Glycoproteins 70:82 Glycoproteins from Seeds of Ginkgo biloba 70:110 Qualitative and Quantitative Analysis of Carbohydrate Modification on Glycoproteins from Seeds of Ginkgo biloba. 28800704 3 71 theme different 636:644 arg1 places 646:651 different places 636:651 different places for detailed structures 636:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 1 72 theme carbohydrate 157:168 arg1 CCDs 199:202 CCDs 199:202 CCDs 199:202 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 1 72 theme carbohydrate 157:168 arg1 determinants 185:196 carbohydrate cross-reactive determinants 157:196 carbohydrate cross-reactive determinants (CCDs) 157:203 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800704 3 73 from moieties 545:552 arg1 seeds 613:617 Ginkgo seeds 606:617 Ginkgo seeds originating from different places for detailed structures 606:675 The aim of this study was to analyze carbohydrate moieties, especially N-linked glycans, of glycoproteins from Ginkgo seeds originating from different places for detailed structures, to enable further research on the role played by N-glycans in Ginkgo-caused allergy. 28800704 1 74 theme moieties 269:276 arg1 importance 242:251 the importance 238:251 the importance of carbohydrate moieties in the innate immune system 238:304 Recent progress in the relationship between carbohydrate cross-reactive determinants (CCDs) and allergic response highlights the importance of carbohydrate moieties in the innate immune system. 28800497 1 0 theme N-glycan 200:207 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 5 1 theme novel 835:839 arg1 glycoconjugates 841:855 These novel glycoconjugates 829:855 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure 829:921 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 4 2 from specificity 750:760 arg1 purification 765:776 purification 765:776 purification of AAL 765:783 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 4 3 theme chromatography 640:653 arg1 column 655:660 An affinity chromatography column 628:660 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide 628:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 1 4 theme trisaccharide 209:221 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 5 5 theme N-glycan 904:911 arg1 structure 913:921 the precise and simplified core-fucose N-glycan structure 865:921 the precise and simplified core-fucose N-glycan structure 865:921 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 4 6 theme affinity 631:638 arg1 column 655:660 An affinity chromatography column 628:660 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide 628:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 4 7 theme good 745:748 arg1 specificity 750:760 a good specificity 743:760 a good specificity in purification of AAL 743:783 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 3 8 contain carrying 592:599 arg1 PAMAM 586:590 the FITC-labeled PAMAM 569:590 the FITC-labeled PAMAM carrying core-fucose trisaccharide 569:625 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 3 8 contain carrying 592:599 arg2 trisaccharide 613:625 core-fucose trisaccharide 601:625 core-fucose trisaccharide 601:625 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 4 9 theme AAL 781:783 arg1 purification 765:776 purification 765:776 purification of AAL 765:783 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 3 10 theme lectin 526:531 arg1 AAL 533:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 1 11 theme GlcNAcβ1,4 223:232 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 3 12 theme model 513:517 arg1 AAL 533:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 3 13 theme binding 547:553 arg1 activity 555:562 binding activity 547:562 binding activity 547:562 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 5 14 theme core-fucose-specific 960:979 arg1 discovery 988:996 core-fucose-specific lectin discovery 960:996 core-fucose-specific lectin discovery 960:996 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 5 15 theme potential 934:942 arg1 application 944:954 a potential application 932:954 a potential application for core-fucose-specific lectin discovery 932:996 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 4 16 contain carrying 693:700 arg1 beads 687:691 the agarose beads 675:691 the agarose beads carrying core-fucosylated trisaccharide 675:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 4 16 contain carrying 693:700 arg2 trisaccharide 719:731 core-fucosylated trisaccharide 702:731 core-fucosylated trisaccharide 702:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 1 17 theme Fucα1,6 234:240 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 0 18 theme Glycoconjugate 0:13 arg1 probes 15:20 Glycoconjugate probes 0:20 Glycoconjugate probes 0:20 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 1 19 theme monosaccharide 346:359 arg1 analysis 361:368 monosaccharide analysis 346:368 monosaccharide analysis with HPAEC-PAD technique 346:393 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 2 20 theme lectin 474:479 arg1 detection 481:489 fucose-specific lectin detection 458:489 fucose-specific lectin detection 458:489 These glycoconjugates as fucose lectin probes were applied in fucose-specific lectin detection and purification. 28800497 4 21 theme agarose 679:685 arg1 beads 687:691 the agarose beads 675:691 the agarose beads carrying core-fucosylated trisaccharide 675:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 5 22 theme precise 869:875 arg1 structure 913:921 the precise and simplified core-fucose N-glycan structure 865:921 the precise and simplified core-fucose N-glycan structure 865:921 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 1 23 with analysis 361:368 arg1 technique 385:393 HPAEC-PAD technique 375:393 HPAEC-PAD technique 375:393 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 1 24 theme Glyco-PAMAM 126:136 arg1 dendrimers 138:147 Glyco-PAMAM dendrimers 126:147 Glyco-PAMAM dendrimers 126:147 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 4 25 theme core-fucosylated 702:717 arg1 trisaccharide 719:731 core-fucosylated trisaccharide 702:731 core-fucosylated trisaccharide 702:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 3 26 theme FITC-labeled 573:584 arg1 PAMAM 586:590 the FITC-labeled PAMAM 569:590 the FITC-labeled PAMAM carrying core-fucose trisaccharide 569:625 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 0 27 theme N-glycan 52:59 arg1 trisaccharide 61:73 a core-fucosylated N-glycan trisaccharide 33:73 a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification 33:123 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 1 28 theme HPAEC-PAD 375:383 arg1 technique 385:393 HPAEC-PAD technique 375:393 HPAEC-PAD technique 375:393 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 5 29 theme simplified 881:890 arg1 structure 913:921 the precise and simplified core-fucose N-glycan structure 865:921 the precise and simplified core-fucose N-glycan structure 865:921 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 4 30 theme non-fucose 790:799 arg1 beads 822:826 non-fucose disaccharide agarose beads 790:826 non-fucose disaccharide agarose beads 790:826 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 4 31 theme disaccharide 801:812 arg1 beads 822:826 non-fucose disaccharide agarose beads 790:826 non-fucose disaccharide agarose beads 790:826 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 0 32 theme core-fucosylated 35:50 arg1 trisaccharide 61:73 a core-fucosylated N-glycan trisaccharide 33:73 a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification 33:123 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 3 33 theme fucose 519:524 arg1 AAL 533:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL 509:535 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 1 34 theme non-fucose 254:263 arg1 GlcNAcβ1,4GlcNAc 278:293 a non-fucose disaccharide GlcNAcβ1,4GlcNAc 252:293 a non-fucose disaccharide GlcNAcβ1,4GlcNAc 252:293 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 1 35 gly core-fucosylated 183:198 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 3 36 theme core-fucose 601:611 arg1 trisaccharide 613:625 core-fucose trisaccharide 601:625 core-fucose trisaccharide 601:625 The model fucose lectin AAL indicated binding activity with the FITC-labeled PAMAM carrying core-fucose trisaccharide. 28800497 5 37 theme core-fucose 892:902 arg1 structure 913:921 the precise and simplified core-fucose N-glycan structure 865:921 the precise and simplified core-fucose N-glycan structure 865:921 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 1 38 theme glyco-agarose 153:165 arg1 beads 167:171 glyco-agarose beads 153:171 glyco-agarose beads 153:171 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 0 39 gly core-fucosylated 35:50 arg1 trisaccharide 61:73 a core-fucosylated N-glycan trisaccharide 33:73 a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification 33:123 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 4 40 gly core-fucosylated 702:717 arg1 trisaccharide 719:731 core-fucosylated trisaccharide 702:731 core-fucosylated trisaccharide 702:731 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 2 41 theme fucose 421:426 arg1 probes 435:440 fucose lectin probes 421:440 fucose lectin probes 421:440 These glycoconjugates as fucose lectin probes were applied in fucose-specific lectin detection and purification. 28800497 1 42 theme disaccharide 265:276 arg1 GlcNAcβ1,4GlcNAc 278:293 a non-fucose disaccharide GlcNAcβ1,4GlcNAc 252:293 a non-fucose disaccharide GlcNAcβ1,4GlcNAc 252:293 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 28800497 5 43 theme lectin 981:986 arg1 discovery 988:996 core-fucose-specific lectin discovery 960:996 core-fucose-specific lectin discovery 960:996 These novel glycoconjugates bearing the precise and simplified core-fucose N-glycan structure provided a potential application for core-fucose-specific lectin discovery. 28800497 0 44 theme lectin 86:91 arg1 identification 93:106 fucose lectin identification 79:106 fucose lectin identification 79:106 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 2 45 theme lectin 428:433 arg1 probes 435:440 fucose lectin probes 421:440 fucose lectin probes 421:440 These glycoconjugates as fucose lectin probes were applied in fucose-specific lectin detection and purification. 28800497 2 46 theme fucose-specific 458:472 arg1 detection 481:489 fucose-specific lectin detection 458:489 fucose-specific lectin detection 458:489 These glycoconjugates as fucose lectin probes were applied in fucose-specific lectin detection and purification. 28800497 4 47 theme agarose 814:820 arg1 beads 822:826 non-fucose disaccharide agarose beads 790:826 non-fucose disaccharide agarose beads 790:826 An affinity chromatography column stuffed with the agarose beads carrying core-fucosylated trisaccharide exhibited a good specificity in purification of AAL than non-fucose disaccharide agarose beads. 28800497 0 48 theme fucose 79:84 arg1 identification 93:106 fucose lectin identification 79:106 fucose lectin identification 79:106 Glycoconjugate probes containing a core-fucosylated N-glycan trisaccharide for fucose lectin identification and purification. 28800497 1 49 theme core-fucosylated 183:198 arg1 GlcNAc 242:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc 181:247 Glyco-PAMAM dendrimers and glyco-agarose beads bearing a core-fucosylated N-glycan trisaccharide GlcNAcβ1,4(Fucα1,6)GlcNAc or a non-fucose disaccharide GlcNAcβ1,4GlcNAc were successfully synthesized and characterized by monosaccharide analysis with HPAEC-PAD technique. 26489654 8 0 theme multifactorial 1355:1368 arg1 mechanism 1370:1378 This intrinsic multifactorial mechanism 1340:1378 This intrinsic multifactorial mechanism of secreted protein aging and turnover 1340:1417 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 6 1 theme distinct 928:935 arg1 receptor 1035:1042 macrophage mannose receptor 1016:1042 macrophage mannose receptor 1016:1042 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 1 theme distinct 928:935 arg1 specificities 945:957 distinct binding specificities 928:957 distinct binding specificities 928:957 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 1 theme distinct 928:935 arg1 αM 1008:1009 integrin αM 999:1009 integrin αM 999:1009 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 1 theme distinct 928:935 arg1 receptor 989:996 the Ashwell-Morell receptor 970:996 the Ashwell-Morell receptor 970:996 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 2 theme secreted 1108:1115 arg1 proteins 1117:1124 secreted proteins 1108:1124 secreted proteins 1108:1124 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 3 theme macrophage 1016:1025 arg1 receptor 1035:1042 macrophage mannose receptor 1016:1042 macrophage mannose receptor 1016:1042 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 7 4 theme N-glycan 1240:1247 arg1 remodeling 1249:1258 N-glycan remodeling 1240:1258 N-glycan remodeling 1240:1258 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 4 5 theme stepwise 528:535 arg1 elimination 537:547 the stepwise elimination 524:547 the stepwise elimination of saccharides attached to the termini of N-glycans 524:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 7 6 theme lectin 1154:1159 arg1 deficiencies 1161:1172 lectin deficiencies 1154:1172 lectin deficiencies 1154:1172 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 7 7 theme distinct 1273:1280 arg1 proteins 1282:1289 distinct proteins 1273:1289 distinct proteins 1273:1289 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 4 8 theme N-glycans 591:599 arg1 termini 580:586 the termini 576:586 the termini of N-glycans 576:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 7 9 theme basal 1226:1230 arg1 rate 1232:1235 the basal rate 1222:1235 the basal rate of N-glycan remodeling 1222:1258 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 7 10 dep life 1328:1331 arg1 spans 1333:1337 spans 1333:1337 spans 1333:1337 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 7 11 theme Glycosidase 1127:1137 arg1 inhibition 1139:1148 Glycosidase inhibition 1127:1148 Glycosidase inhibition 1127:1148 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 8 12 theme secreted 1383:1390 arg1 protein 1392:1398 secreted protein aging and turnover 1383:1417 secreted protein aging and turnover 1383:1417 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 3 13 mod modified 381:388 arg1 proteins 348:355 Almost all secreted proteins 328:355 Almost all secreted proteins 328:355 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 3 13 mod modified 381:388 arg3 attachment 408:417 the covalent attachment 395:417 the covalent attachment of N-glycans 395:430 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 6 14 theme endocytic 905:913 arg1 lectins 915:921 endocytic lectins 905:921 endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, 905:1043 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 8 15 theme protein 1392:1398 arg1 mechanism 1370:1378 This intrinsic multifactorial mechanism 1340:1378 This intrinsic multifactorial mechanism of secreted protein aging and turnover 1340:1417 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 7 16 theme protein 1184:1190 arg1 life 1192:1195 protein life spans 1184:1201 protein life spans 1184:1201 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 3 17 theme secreted 339:346 arg1 proteins 348:355 Almost all secreted proteins 328:355 Almost all secreted proteins 328:355 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 1 18 theme secreted 100:107 arg1 proteome 109:116 the secreted proteome 96:116 the secreted proteome 96:116 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 8 19 dep protein 1392:1398 arg1 turnover 1410:1417 turnover 1410:1417 turnover 1410:1417 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 8 19 dep protein 1392:1398 arg1 aging 1400:1404 aging 1400:1404 aging 1400:1404 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 4 20 theme turnover 505:512 arg1 mechanism 465:473 an intrinsic mechanism 452:473 an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans 452:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 0 21 theme protein 35:41 arg1 aging 43:47 secreted protein aging 26:47 secreted protein aging 26:47 An intrinsic mechanism of secreted protein aging and turnover. 26489654 5 22 theme N-glycan 797:804 arg1 structures 806:815 N-glycan structures 797:815 N-glycan structures 797:815 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 1 23 theme different 154:162 arg1 proteins 164:171 different proteins 154:171 different proteins 154:171 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 0 24 theme intrinsic 3:11 arg1 mechanism 13:21 An intrinsic mechanism 0:21 An intrinsic mechanism of secreted protein aging and turnover 0:60 An intrinsic mechanism of secreted protein aging and turnover. 26489654 4 25 dep protein 487:493 arg1 aging 495:499 aging 495:499 aging 495:499 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 1 26 dep life 140:143 arg1 spans 145:149 spans 145:149 spans of different proteins 145:171 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 4 27 theme protein 487:493 arg1 mechanism 465:473 an intrinsic mechanism 452:473 an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans 452:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 5 28 theme protein 878:884 arg1 age 886:888 increased protein age 868:888 increased protein age 868:888 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 3 29 theme covalent 399:406 arg1 attachment 408:417 the covalent attachment 395:417 the covalent attachment of N-glycans 395:430 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 6 30 theme integrin 999:1006 arg1 αM 1008:1009 integrin αM 999:1009 integrin αM 999:1009 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 7 31 dep life 1192:1195 arg1 spans 1197:1201 spans 1197:1201 spans 1197:1201 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 5 32 theme Endogenous 602:611 arg1 beta-galactosidase 685:702 beta-galactosidase 1 685:704 beta-galactosidase 1 (Glb1) 685:711 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 32 theme Endogenous 602:611 arg1 B 733:733 hexosaminidase B 718:733 hexosaminidase B (HexB) 718:740 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 32 theme Endogenous 602:611 arg1 glycosidases 613:624 Endogenous glycosidases 602:624 Endogenous glycosidases 602:624 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 32 theme Endogenous 602:611 arg1 neuraminidase 637:649 neuraminidase 1 637:651 neuraminidase 1 (Neu1) 637:658 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 32 theme Endogenous 602:611 arg1 neuraminidase 661:673 neuraminidase 3 661:675 neuraminidase 3 (Neu3) 661:682 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 6 33 theme Ashwell-Morell 974:987 arg1 receptor 989:996 the Ashwell-Morell receptor 970:996 the Ashwell-Morell receptor 970:996 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 7 34 from differences 1307:1317 arg1 life 1328:1331 their life 1322:1331 their life spans 1322:1337 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 7 35 dep proteins 1282:1289 arg1 accounting 1292:1301 accounting 1292:1301 accounting for differences in their life spans 1292:1337 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 6 36 theme ligand 1069:1074 arg1 recognition 1076:1086 N-glycan ligand recognition 1060:1086 N-glycan ligand recognition 1060:1086 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 4 37 attach attached 564:571 arg1 termini 580:586 the termini 576:586 the termini of N-glycans 576:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 4 37 attach attached 564:571 arg2 saccharides 552:562 saccharides 552:562 saccharides attached to the termini of N-glycans 552:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 6 38 theme N-glycan 1060:1067 arg1 recognition 1076:1086 N-glycan ligand recognition 1060:1086 N-glycan ligand recognition 1060:1086 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 3 39 theme N-glycans 422:430 arg1 attachment 408:417 the covalent attachment 395:417 the covalent attachment of N-glycans 395:430 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 1 40 dep composition 67:77 arg1 The 63:65 The 63:65 The 63:65 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 4 41 theme intrinsic 455:463 arg1 mechanism 465:473 an intrinsic mechanism 452:473 an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans 452:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 2 42 theme intrinsic 218:226 arg1 factors 228:234 intrinsic factors 218:234 intrinsic factors determining secreted protein aging and turnover 218:282 However, unlike intracellular protein fate, intrinsic factors determining secreted protein aging and turnover have not been identified and characterized. 26489654 5 43 theme different 841:849 arg1 saccharides 851:861 different saccharides 841:861 different saccharides 841:861 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 0 44 theme secreted 26:33 arg1 aging 43:47 secreted protein aging 26:47 secreted protein aging 26:47 An intrinsic mechanism of secreted protein aging and turnover. 26489654 4 45 theme secreted 478:485 arg1 protein 487:493 secreted protein aging and turnover 478:512 protein 487:493 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 0 46 theme turnover 53:60 arg1 mechanism 13:21 An intrinsic mechanism 0:21 An intrinsic mechanism of secreted protein aging and turnover 0:60 An intrinsic mechanism of secreted protein aging and turnover. 26489654 4 47 attach linked 514:519 arg2 protein 487:493 secreted protein aging and turnover 478:512 protein 487:493 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 4 47 attach linked 514:519 arg1 elimination 537:547 the stepwise elimination 524:547 the stepwise elimination of saccharides attached to the termini of N-glycans 524:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 0 48 theme aging 43:47 arg1 mechanism 13:21 An intrinsic mechanism 0:21 An intrinsic mechanism of secreted protein aging and turnover 0:60 An intrinsic mechanism of secreted protein aging and turnover. 26489654 6 49 theme binding 937:943 arg1 receptor 1035:1042 macrophage mannose receptor 1016:1042 macrophage mannose receptor 1016:1042 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 49 theme binding 937:943 arg1 specificities 945:957 distinct binding specificities 928:957 distinct binding specificities 928:957 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 49 theme binding 937:943 arg1 αM 1008:1009 integrin αM 999:1009 integrin αM 999:1009 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 49 theme binding 937:943 arg1 receptor 989:996 the Ashwell-Morell receptor 970:996 the Ashwell-Morell receptor 970:996 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 2 50 theme intracellular 190:202 arg1 fate 212:215 intracellular protein fate 190:215 intracellular protein fate 190:215 However, unlike intracellular protein fate, intrinsic factors determining secreted protein aging and turnover have not been identified and characterized. 26489654 2 51 theme protein 204:210 arg1 fate 212:215 intracellular protein fate 190:215 intracellular protein fate 190:215 However, unlike intracellular protein fate, intrinsic factors determining secreted protein aging and turnover have not been identified and characterized. 26489654 1 52 theme proteome 109:116 arg1 functions 83:91 functions 83:91 functions 83:91 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 1 52 theme proteome 109:116 arg1 composition 67:77 composition 67:77 composition 67:77 The composition and functions of the secreted proteome are controlled by the life spans of different proteins. 26489654 4 53 theme saccharides 552:562 arg1 elimination 537:547 the stepwise elimination 524:547 the stepwise elimination of saccharides attached to the termini of N-glycans 524:599 We have discovered an intrinsic mechanism of secreted protein aging and turnover linked to the stepwise elimination of saccharides attached to the termini of N-glycans. 26489654 6 54 with lectins 915:921 arg1 receptor 1035:1042 macrophage mannose receptor 1016:1042 macrophage mannose receptor 1016:1042 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 54 with lectins 915:921 arg1 specificities 945:957 distinct binding specificities 928:957 distinct binding specificities 928:957 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 54 with lectins 915:921 arg1 αM 1008:1009 integrin αM 999:1009 integrin αM 999:1009 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 54 with lectins 915:921 arg1 receptor 989:996 the Ashwell-Morell receptor 970:996 the Ashwell-Morell receptor 970:996 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 8 55 theme disease 1461:1467 arg1 health 1434:1439 health 1434:1439 health 1434:1439 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 8 55 theme disease 1461:1467 arg1 outcomes 1449:1456 the outcomes 1445:1456 the outcomes of disease 1445:1467 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 2 56 theme protein 257:263 arg1 aging 265:269 secreted protein aging 248:269 secreted protein aging 248:269 However, unlike intracellular protein fate, intrinsic factors determining secreted protein aging and turnover have not been identified and characterized. 26489654 5 57 theme hydrolytic 751:760 arg1 activities 762:771 hydrolytic activities 751:771 hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age 751:888 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 2 58 theme secreted 248:255 arg1 aging 265:269 secreted protein aging 248:269 secreted protein aging 248:269 However, unlike intracellular protein fate, intrinsic factors determining secreted protein aging and turnover have not been identified and characterized. 26489654 7 59 theme remodeling 1249:1258 arg1 rate 1232:1235 the basal rate 1222:1235 the basal rate of N-glycan remodeling 1222:1258 Glycosidase inhibition and lectin deficiencies increased protein life spans and abundance, and the basal rate of N-glycan remodeling varied among distinct proteins, accounting for differences in their life spans. 26489654 8 60 theme intrinsic 1345:1353 arg1 mechanism 1370:1378 This intrinsic multifactorial mechanism 1340:1378 This intrinsic multifactorial mechanism of secreted protein aging and turnover 1340:1417 This intrinsic multifactorial mechanism of secreted protein aging and turnover contributes to health and the outcomes of disease. 26489654 5 61 theme increased 868:876 arg1 age 886:888 increased protein age 868:888 increased protein age 868:888 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 3 62 theme all 335:337 arg1 proteins 348:355 Almost all secreted proteins 328:355 Almost all secreted proteins 328:355 Almost all secreted proteins are posttranslationally modified with the covalent attachment of N-glycans. 26489654 5 63 contain possess 743:749 arg1 glycosidases 613:624 Endogenous glycosidases 602:624 Endogenous glycosidases 602:624 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 63 contain possess 743:749 arg2 activities 762:771 hydrolytic activities 751:771 hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age 751:888 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 63 contain possess 743:749 arg1 beta-galactosidase 685:702 beta-galactosidase 1 685:704 beta-galactosidase 1 (Glb1) 685:711 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 63 contain possess 743:749 arg1 neuraminidase 637:649 neuraminidase 1 637:651 neuraminidase 1 (Neu1) 637:658 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 63 contain possess 743:749 arg1 neuraminidase 661:673 neuraminidase 3 661:675 neuraminidase 3 (Neu3) 661:682 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 63 contain possess 743:749 arg1 B 733:733 hexosaminidase B 718:733 hexosaminidase B (HexB) 718:740 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 6 64 theme mannose 1027:1033 arg1 receptor 1035:1042 macrophage mannose receptor 1016:1042 macrophage mannose receptor 1016:1042 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 65 theme proteins 1117:1124 arg1 recognition 1076:1086 N-glycan ligand recognition 1060:1086 N-glycan ligand recognition 1060:1086 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 6 65 theme proteins 1117:1124 arg1 turnover 1096:1103 the turnover 1092:1103 the turnover of secreted proteins 1092:1124 Subsequently, endocytic lectins with distinct binding specificities, including the Ashwell-Morell receptor, integrin αM, and macrophage mannose receptor, are engaged in N-glycan ligand recognition and the turnover of secreted proteins. 26489654 5 66 theme hexosaminidase 718:731 arg1 B 733:733 hexosaminidase B 718:733 hexosaminidase B (HexB) 718:740 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 26489654 5 66 theme hexosaminidase 718:731 arg1 HexB 736:739 HexB 736:739 HexB 736:739 Endogenous glycosidases, including neuraminidase 1 (Neu1), neuraminidase 3 (Neu3), beta-galactosidase 1 (Glb1), and hexosaminidase B (HexB), possess hydrolytic activities that temporally remodel N-glycan structures, progressively exposing different saccharides with increased protein age. 27585995 15 0 theme diseases 3489:3496 arg1 biomarkers 3464:3473 O-glycan biomarkers 3455:3473 O-glycan biomarkers of some major diseases 3455:3496 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 0 theme diseases 3489:3496 arg1 targets 3519:3525 O-glycan related targets 3502:3525 O-glycan related targets of some drugs 3502:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 4 1 theme chromophoric 1111:1122 arg1 labeling 1124:1131 in situ chromophoric labeling 1103:1131 in situ chromophoric labeling of intact mucin-type O-glycans in one step 1103:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 3 2 theme mass 730:733 arg1 difference 735:744 the too large or too small inconstant mass difference 692:744 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 1 3 from studies 349:355 arg1 patterns 384:391 protein O-glycosylation patterns 360:391 protein O-glycosylation patterns 360:391 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 1 3 from studies 349:355 arg1 search 401:406 the search 397:406 the search for disease O-glycan biomarkers 397:438 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 1 4 theme quantitative 267:278 arg1 analysis 280:287 quantitative analysis 267:287 quantitative analysis of glycoprotein O-glycans 267:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 11 5 theme nascent 2559:2565 arg1 proteins 2567:2574 nascent proteins 2559:2574 nascent proteins 2559:2574 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 10 6 theme quantitative 2201:2212 arg1 comparison 2225:2234 the quantitative O-glycomic comparison 2197:2234 the quantitative O-glycomic comparison between perch and salmon eggs 2197:2264 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 3 7 theme heavy 768:772 arg1 derivatives 787:797 the light and heavy isotope form derivatives 754:797 the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 754:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 0 8 theme mass 198:201 arg1 analysis 217:224 mass spectrometric analysis 198:224 mass spectrometric analysis 198:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 13 9 theme important 2907:2915 arg1 opportunity 2917:2927 an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans 2904:2998 an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans 2904:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 12 10 theme informational 2648:2660 arg1 molecule 2662:2669 informational molecule 2648:2669 informational molecule 2648:2669 As a type of informational molecule, the O-glycans of these glycoproteins participate directly in these biological mechanisms. 27585995 3 11 theme light 758:762 arg1 derivatives 787:797 the light and heavy isotope form derivatives 754:797 the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 754:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 4 12 dep in 1103:1104 arg1 situ 1106:1109 situ 1106:1109 situ 1106:1109 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 11 13 dep serine 2515:2520 arg1 residues 2547:2554 residues 2547:2554 residues of nascent proteins 2547:2574 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 11 13 dep serine 2515:2520 arg1 proteins 2567:2574 nascent proteins 2559:2574 nascent proteins 2559:2574 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 7 14 theme strong 1731:1736 arg1 adsorption 1741:1750 a strong UV adsorption 1729:1750 a strong UV adsorption 1729:1750 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 4 15 theme relative 976:983 arg1 strategy 1000:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 11 16 theme existing 2468:2475 arg1 process 2504:2510 a widely existing glycosylation modification process 2459:2510 a widely existing glycosylation modification process 2459:2510 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 3 17 theme form 782:785 arg1 derivatives 787:797 the light and heavy isotope form derivatives 754:797 the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 754:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 0 18 theme isotope 126:132 arg1 labeling 134:141 simultaneous stable isotope labeling 106:141 simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 106:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 1 19 theme disease 412:418 arg1 biomarkers 429:438 disease O-glycan biomarkers 412:438 disease O-glycan biomarkers 412:438 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 6 20 theme spectrometric 1595:1607 arg1 techniques 1609:1618 mass spectrometric techniques 1590:1618 mass spectrometric techniques 1590:1618 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 15 21 theme quantitative/comparative 3287:3310 arg1 O-glycomics 3312:3322 quantitative/comparative O-glycomics 3287:3322 quantitative/comparative O-glycomics 3287:3322 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 10 22 theme O-glycomic 2214:2223 arg1 comparison 2225:2234 the quantitative O-glycomic comparison 2197:2234 the quantitative O-glycomic comparison between perch and salmon eggs 2197:2264 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 9 23 theme bovine 2068:2073 arg1 fetuin 2075:2080 bovine fetuin and porcine stomach mucin 2068:2106 fetuin 2075:2080 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 6 24 theme obtained 1377:1384 arg1 derivatives 1395:1405 The obtained O-glycan derivatives 1373:1405 The obtained O-glycan derivatives 1373:1405 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 3 25 theme O-glycan 656:663 arg1 methods 673:679 reliable nonreductive O-glycan release methods 634:679 reliable nonreductive O-glycan release methods 634:679 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 4 26 theme facile 946:951 arg1 strategy 1000:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 11 27 theme essential 2582:2590 arg1 roles 2592:2596 essential roles 2582:2596 essential roles 2582:2596 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 15 28 theme biomarkers 3464:3473 arg1 search 3445:3450 the search 3441:3450 the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs 3441:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 14 29 theme versatile 3058:3066 arg1 method 3079:3084 a simple and versatile analytical method 3045:3084 a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry 3045:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 3 30 theme reliable 634:641 arg1 methods 673:679 reliable nonreductive O-glycan release methods 634:679 reliable nonreductive O-glycan release methods 634:679 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 14 31 theme mass 3144:3147 arg1 spectrometry 3149:3160 mass spectrometry 3144:3160 mass spectrometry 3144:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 3 32 theme mass 908:911 arg1 spectrometry 913:924 mass spectrometry 908:924 mass spectrometry 908:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 7 33 theme high 1698:1701 arg1 hydrophobicity 1703:1716 a relatively high hydrophobicity 1685:1716 a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV 1685:1847 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 4 34 theme versatile 957:965 arg1 strategy 1000:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 9 35 theme stomach 2094:2100 arg1 mucin 2102:2106 bovine fetuin and porcine stomach mucin 2068:2106 mucin 2102:2106 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 14 36 theme simple 3047:3052 arg1 method 3079:3084 a simple and versatile analytical method 3045:3084 a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry 3045:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 8 37 theme one-pot 1889:1895 arg1 reaction 1897:1904 the one-pot reaction 1885:1904 the one-pot reaction as well as the corresponding sample purification approach 1885:1962 We have refined the conditions for the one-pot reaction as well as the corresponding sample purification approach. 27585995 3 38 theme O-glycans 895:903 arg1 recognition 854:864 recognition 854:864 recognition 854:864 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 3 38 theme O-glycans 895:903 arg1 analysis 883:890 quantitative analysis 870:890 quantitative analysis 870:890 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 15 39 theme complex 3372:3378 arg1 relationship 3399:3410 the complex structure-function relationship 3368:3410 the complex structure-function relationship of O-glycans 3368:3423 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 4 40 theme one-pot 1031:1037 arg1 method 1039:1044 an improved one-pot method 1019:1044 an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step 1019:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 10 41 theme excellent 2334:2342 arg1 applicability 2344:2356 its excellent applicability 2330:2356 its excellent applicability to various complex biological samples 2330:2394 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 0 42 theme method 61:66 arg1 improvement 34:44 improvement 34:44 improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 34:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 3 43 theme methods 673:679 arg1 difference 735:744 the too large or too small inconstant mass difference 692:744 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 3 43 theme methods 673:679 arg1 shortage 622:629 the shortage 618:629 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 14 44 from changes 3211:3217 arg1 level 3246:3250 expression level 3235:3250 expression level 3235:3250 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 0 45 theme nonreductive 72:83 arg1 release 94:100 nonreductive O-glycan release 72:100 nonreductive O-glycan release 72:100 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 1 46 theme O-glycosylation 368:382 arg1 patterns 384:391 protein O-glycosylation patterns 360:391 protein O-glycosylation patterns 360:391 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 9 47 gly O-glycoproteins 2117:2131 arg1 O-glycoproteins 2117:2131 model O-glycoproteins 2111:2131 model O-glycoproteins 2111:2131 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 12 48 gly glycoproteins 2695:2707 arg1 glycoproteins 2695:2707 these glycoproteins 2689:2707 these glycoproteins 2689:2707 As a type of informational molecule, the O-glycans of these glycoproteins participate directly in these biological mechanisms. 27585995 11 49 theme BIOLOGICAL 2397:2406 arg1 glycoproteins 2430:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 11 50 theme biological 2613:2622 arg1 processes 2624:2632 biological processes 2613:2632 biological processes 2613:2632 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 13 51 theme various 2969:2975 arg1 O-glycans 2990:2998 various glycoprotein O-glycans 2969:2998 various glycoprotein O-glycans 2969:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 10 52 theme complex 2369:2375 arg1 samples 2388:2394 various complex biological samples 2361:2394 various complex biological samples 2361:2394 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 6 53 theme 10-Da 1427:1431 arg1 difference 1438:1447 a permanent 10-Da mass difference 1415:1447 a permanent 10-Da mass difference between the d0- and d5-PMP forms 1415:1480 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 8 54 theme corresponding 1921:1933 arg1 approach 1955:1962 the corresponding sample purification approach 1917:1962 the one-pot reaction as well as the corresponding sample purification approach 1885:1962 We have refined the conditions for the one-pot reaction as well as the corresponding sample purification approach. 27585995 7 55 theme high-resolution 1777:1791 arg1 separation 1793:1802 high-resolution separation 1777:1802 high-resolution separation 1777:1802 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 7 56 theme d5-PMP 1643:1648 arg1 derivatives 1650:1660 the d0- and d5-PMP derivatives 1631:1660 derivatives 1650:1660 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 2 57 theme promising 570:578 arg1 technique 580:588 an ideal and promising technique 557:588 an ideal and promising technique 557:588 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 0 58 theme simultaneous 106:117 arg1 labeling 134:141 simultaneous stable isotope labeling 106:141 simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 106:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 8 59 theme purification 1942:1953 arg1 approach 1955:1962 the corresponding sample purification approach 1917:1962 the one-pot reaction as well as the corresponding sample purification approach 1885:1962 We have refined the conditions for the one-pot reaction as well as the corresponding sample purification approach. 27585995 7 60 theme suitable 1764:1771 arg1 adsorption 1741:1750 a strong UV adsorption 1729:1750 a strong UV adsorption 1729:1750 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 4 61 from release 1091:1097 arg1 step 1171:1174 one step 1167:1174 one step 1167:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 2 62 theme ideal 560:564 arg1 technique 580:588 an ideal and promising technique 557:588 an ideal and promising technique 557:588 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 0 63 theme Quantitative 0:11 arg1 O-glycomics 13:23 Quantitative O-glycomics 0:23 Quantitative O-glycomics 0:23 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 1 64 theme glycoprotein 292:303 arg1 O-glycans 305:313 glycoprotein O-glycans 292:313 glycoprotein O-glycans 292:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 13 65 theme functional 2944:2953 arg1 mechanisms 2955:2964 the functional mechanisms 2940:2964 the functional mechanisms of various glycoprotein O-glycans 2940:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 13 66 from differences 2787:2797 arg1 level 2837:2841 expression level 2826:2841 expression level 2826:2841 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 2 67 theme stable 482:487 arg1 labeling 497:504 stable isotope labeling 482:504 stable isotope labeling followed by mass spectrometric analysis 482:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 14 68 theme O-glycans 3222:3230 arg1 evaluation 3178:3187 rapid evaluation 3172:3187 rapid evaluation of the differences 3172:3206 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 14 68 theme O-glycans 3222:3230 arg1 changes 3211:3217 changes 3211:3217 changes of O-glycans in expression level 3211:3250 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 15 69 contain has 3331:3333 arg2 significance 3341:3352 great significance 3335:3352 great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs 3335:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 69 contain has 3331:3333 arg1 O-glycomics 3312:3322 quantitative/comparative O-glycomics 3287:3322 quantitative/comparative O-glycomics 3287:3322 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 2 70 theme O-glycans 466:474 arg1 quantitation 450:461 Relative quantitation 441:461 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis 441:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 5 71 theme quantitative 1240:1251 arg1 release 1262:1268 quantitative O-glycan release 1240:1268 quantitative O-glycan release 1240:1268 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 11 72 theme proteins 2567:2574 arg1 residues 2547:2554 residues 2547:2554 residues of nascent proteins 2547:2574 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 11 72 theme proteins 2567:2574 arg1 proteins 2567:2574 nascent proteins 2559:2574 nascent proteins 2559:2574 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 7 73 theme UV 1738:1739 arg1 adsorption 1741:1750 a strong UV adsorption 1729:1750 a strong UV adsorption 1729:1750 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 2 74 theme spectrometric 523:535 arg1 analysis 537:544 mass spectrometric analysis 518:544 mass spectrometric analysis 518:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 3 75 theme large 700:704 arg1 difference 735:744 the too large or too small inconstant mass difference 692:744 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 4 76 theme nonreductive 1078:1089 arg1 release 1091:1097 nonreductive release 1078:1097 nonreductive release 1078:1097 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 1 77 theme current 341:347 arg1 studies 349:355 current studies 341:355 current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers 341:438 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 14 78 theme glycoprotein 3118:3129 arg1 O-glycans 3131:3139 glycoprotein O-glycans 3118:3139 glycoprotein O-glycans 3118:3139 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 15 79 theme related 3511:3517 arg1 targets 3519:3525 O-glycan related targets 3502:3525 O-glycan related targets of some drugs 3502:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 4 80 theme O-glycans 1154:1162 arg1 labeling 1124:1131 in situ chromophoric labeling 1103:1131 in situ chromophoric labeling of intact mucin-type O-glycans in one step 1103:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 4 80 theme O-glycans 1154:1162 arg1 release 1091:1097 nonreductive release 1078:1097 nonreductive release 1078:1097 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 3 81 theme inconstant 719:728 arg1 difference 735:744 the too large or too small inconstant mass difference 692:744 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 6 82 theme O-glycans 1577:1585 arg1 discrimination 1501:1514 complete discrimination 1492:1514 complete discrimination 1492:1514 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 6 82 theme O-glycans 1577:1585 arg1 quantification 1532:1545 comparative quantification 1520:1545 comparative quantification 1520:1545 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 1 83 theme simple 234:239 arg1 methods 255:261 Rapid, simple and versatile methods 227:261 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans 227:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 13 84 theme characteristic 2772:2785 arg1 differences 2787:2797 the characteristic differences 2768:2797 the characteristic differences 2768:2797 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 9 85 theme quantitation 1974:1985 arg1 feasibility 1987:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 11 86 theme O-Linked 2421:2428 arg1 glycoproteins 2430:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 14 87 theme rapid 3172:3176 arg1 evaluation 3178:3187 rapid evaluation 3172:3187 rapid evaluation of the differences 3172:3206 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 1 88 theme versatile 245:253 arg1 methods 255:261 Rapid, simple and versatile methods 227:261 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans 227:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 0 89 theme one-pot 53:59 arg1 method 61:66 the one-pot method 49:66 the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 49:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 7 90 contain have 1680:1683 arg1 derivatives 1650:1660 the d0- and d5-PMP derivatives 1631:1660 derivatives 1650:1660 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 7 90 contain have 1680:1683 arg2 adsorption 1741:1750 a strong UV adsorption 1729:1750 a strong UV adsorption 1729:1750 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 7 90 contain have 1680:1683 arg2 hydrophobicity 1703:1716 a relatively high hydrophobicity 1685:1716 a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV 1685:1847 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 15 91 theme major 3483:3487 arg1 diseases 3489:3496 some major diseases 3478:3496 some major diseases 3478:3496 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 12 92 theme molecule 2662:2669 arg1 type 2640:2643 a type 2638:2643 a type of informational molecule 2638:2669 As a type of informational molecule, the O-glycans of these glycoproteins participate directly in these biological mechanisms. 27585995 13 93 theme diseases 2881:2888 arg1 pathologies 2861:2871 pathologies 2861:2871 pathologies of many diseases 2861:2888 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 15 94 theme O-glycan 3455:3462 arg1 biomarkers 3464:3473 O-glycan biomarkers 3455:3473 O-glycan biomarkers of some major diseases 3455:3496 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 4 95 theme intact 1136:1141 arg1 O-glycans 1154:1162 intact mucin-type O-glycans 1136:1162 intact mucin-type O-glycans 1136:1162 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 0 96 theme spectrometric 203:215 arg1 analysis 217:224 mass spectrometric analysis 198:224 mass spectrometric analysis 198:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 6 97 theme comparative 1520:1530 arg1 quantification 1532:1545 comparative quantification 1520:1545 comparative quantification 1520:1545 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 3 98 theme isotope 774:780 arg1 derivatives 787:797 the light and heavy isotope form derivatives 754:797 the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 754:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 14 99 theme O-glycans 3131:3139 arg1 quantitation 3102:3113 the precise quantitation 3090:3113 the precise quantitation of glycoprotein O-glycans by mass spectrometry 3090:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 14 100 theme differences 3196:3206 arg1 evaluation 3178:3187 rapid evaluation 3172:3187 rapid evaluation of the differences 3172:3206 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 14 100 theme differences 3196:3206 arg1 changes 3211:3217 changes 3211:3217 changes of O-glycans in expression level 3211:3250 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 9 101 theme strategy 2034:2041 arg1 reliability 2000:2010 reliability 2000:2010 reliability 2000:2010 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 9 101 theme strategy 2034:2041 arg1 feasibility 1987:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 9 101 theme strategy 2034:2041 arg1 linearity 2016:2024 linearity 2016:2024 linearity 2016:2024 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 13 102 theme O-glycans 2813:2821 arg1 changes 2802:2808 changes 2802:2808 changes of O-glycans in expression level 2802:2841 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 13 102 theme O-glycans 2813:2821 arg1 differences 2787:2797 the characteristic differences 2768:2797 the characteristic differences 2768:2797 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 15 103 theme O-glycomics 3312:3322 arg1 field 3278:3282 the field 3274:3282 the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs 3274:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 104 theme drugs 3535:3539 arg1 biomarkers 3464:3473 O-glycan biomarkers 3455:3473 O-glycan biomarkers of some major diseases 3455:3496 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 104 theme drugs 3535:3539 arg1 targets 3519:3525 O-glycan related targets 3502:3525 O-glycan related targets of some drugs 3502:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 14 105 theme novel 3005:3009 arg1 strategy 3011:3018 The novel strategy 3001:3018 The novel strategy introduced here 3001:3034 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 3 106 theme O-glycans 802:810 arg1 derivatives 787:797 the light and heavy isotope form derivatives 754:797 the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 754:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 14 107 theme precise 3094:3100 arg1 quantitation 3102:3113 the precise quantitation 3090:3113 the precise quantitation of glycoprotein O-glycans by mass spectrometry 3090:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 0 108 with release 94:100 arg1 phenyl-3-methyl-5-pyrazolone 157:184 1-(d0/d5)phenyl-3-methyl-5-pyrazolone 148:184 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 148:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 13 109 theme expression 2826:2835 arg1 level 2837:2841 expression level 2826:2841 expression level 2826:2841 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 4 110 theme O-glycan 967:974 arg1 strategy 1000:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 6 111 theme O-glycan 1386:1393 arg1 derivatives 1395:1405 The obtained O-glycan derivatives 1373:1405 The obtained O-glycan derivatives 1373:1405 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 1 112 theme O-glycan 420:427 arg1 biomarkers 429:438 disease O-glycan biomarkers 412:438 disease O-glycan biomarkers 412:438 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 3 113 theme nonreductive 643:654 arg1 methods 673:679 reliable nonreductive O-glycan release methods 634:679 reliable nonreductive O-glycan release methods 634:679 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 6 114 theme mass 1590:1593 arg1 techniques 1609:1618 mass spectrometric techniques 1590:1618 mass spectrometric techniques 1590:1618 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 1 115 theme Rapid 227:231 arg1 methods 255:261 Rapid, simple and versatile methods 227:261 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans 227:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 4 116 theme quantification 985:998 arg1 strategy 1000:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 a facile and versatile O-glycan relative quantification strategy 944:1007 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 14 117 theme expression 3235:3244 arg1 level 3246:3250 expression level 3235:3250 expression level 3235:3250 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 14 118 theme analytical 3068:3077 arg1 method 3079:3084 a simple and versatile analytical method 3045:3084 a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry 3045:3160 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 3 119 dep recognition 854:864 arg1 the 850:852 the 850:852 the 850:852 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 1 120 gly glycoprotein 292:303 arg1 glycoprotein 292:303 glycoprotein O-glycans 292:313 glycoprotein O-glycans 292:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 9 121 theme porcine 2086:2092 arg1 mucin 2102:2106 bovine fetuin and porcine stomach mucin 2068:2106 mucin 2102:2106 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 5 122 theme d5- 1330:1332 arg1 1-phenyl-3-methyl-5-pyrazolone 1335:1364 either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone 1287:1364 1-phenyl-3-methyl-5-pyrazolone 1335:1364 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 12 123 theme biological 2739:2748 arg1 mechanisms 2750:2759 these biological mechanisms 2733:2759 these biological mechanisms 2733:2759 As a type of informational molecule, the O-glycans of these glycoproteins participate directly in these biological mechanisms. 27585995 15 124 theme O-glycan 3502:3509 arg1 targets 3519:3525 O-glycan related targets 3502:3525 O-glycan related targets of some drugs 3502:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 0 125 with labeling 134:141 arg1 phenyl-3-methyl-5-pyrazolone 157:184 1-(d0/d5)phenyl-3-methyl-5-pyrazolone 148:184 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 148:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 6 126 theme d0- 1461:1463 arg1 forms 1476:1480 the d0- and d5-PMP forms 1457:1480 forms 1476:1480 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 3 127 theme release 665:671 arg1 methods 673:679 reliable nonreductive O-glycan release methods 634:679 reliable nonreductive O-glycan release methods 634:679 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 9 128 theme good 1969:1972 arg1 feasibility 1987:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility 1965:1997 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 7 129 theme O-glycans 1665:1673 arg1 derivatives 1650:1660 the d0- and d5-PMP derivatives 1631:1660 derivatives 1650:1660 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 4 130 theme improved 1022:1029 arg1 method 1039:1044 an improved one-pot method 1019:1044 an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step 1019:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 11 131 theme processes 2624:2632 arg1 series 2603:2608 a series 2601:2608 a series of biological processes 2601:2632 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 9 132 theme model 2111:2115 arg1 O-glycoproteins 2117:2131 model O-glycoproteins 2111:2131 model O-glycoproteins 2111:2131 The good quantitation feasibility, reliability and linearity of this strategy have been verified using bovine fetuin and porcine stomach mucin as model O-glycoproteins. 27585995 12 133 theme glycoproteins 2695:2707 arg1 O-glycans 2676:2684 the O-glycans 2672:2684 the O-glycans of these glycoproteins 2672:2707 As a type of informational molecule, the O-glycans of these glycoproteins participate directly in these biological mechanisms. 27585995 0 134 theme O-glycan 85:92 arg1 release 94:100 nonreductive O-glycan release 72:100 nonreductive O-glycan release 72:100 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 1 135 theme protein 360:366 arg1 patterns 384:391 protein O-glycosylation patterns 360:391 protein O-glycosylation patterns 360:391 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 4 136 from labeling 1124:1131 arg1 step 1171:1174 one step 1167:1174 one step 1167:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 15 137 theme great 3335:3339 arg1 significance 3341:3352 great significance 3335:3352 great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs 3335:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 138 theme structure-function 3380:3397 arg1 relationship 3399:3410 the complex structure-function relationship 3368:3410 the complex structure-function relationship of O-glycans 3368:3423 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 10 139 theme biological 2377:2386 arg1 samples 2388:2394 various complex biological samples 2361:2394 various complex biological samples 2361:2394 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 5 140 theme deuterated 1318:1327 arg1 1-phenyl-3-methyl-5-pyrazolone 1335:1364 either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone 1287:1364 1-phenyl-3-methyl-5-pyrazolone 1335:1364 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 6 141 theme mass 1433:1436 arg1 difference 1438:1447 a permanent 10-Da mass difference 1415:1447 a permanent 10-Da mass difference between the d0- and d5-PMP forms 1415:1480 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 5 142 theme one-pot 1196:1202 arg1 method 1204:1209 the one-pot method 1192:1209 the one-pot method 1192:1209 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 13 143 theme glycoprotein 2977:2988 arg1 O-glycans 2990:2998 various glycoprotein O-glycans 2969:2998 various glycoprotein O-glycans 2969:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 7 144 theme high-sensitivity 1808:1823 arg1 detection 1825:1833 high-sensitivity detection 1808:1833 high-sensitivity detection 1808:1833 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 11 145 theme SIGNIFICANCE 2408:2419 arg1 glycoproteins 2430:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 10 146 theme various 2361:2367 arg1 samples 2388:2394 various complex biological samples 2361:2394 various complex biological samples 2361:2394 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 3 147 theme quantitative 870:881 arg1 analysis 883:890 quantitative analysis 870:890 quantitative analysis 870:890 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 6 148 theme permanent 1417:1425 arg1 difference 1438:1447 a permanent 10-Da mass difference 1415:1447 a permanent 10-Da mass difference between the d0- and d5-PMP forms 1415:1480 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 13 149 theme O-glycans 2990:2998 arg1 mechanisms 2955:2964 the functional mechanisms 2940:2964 the functional mechanisms of various glycoprotein O-glycans 2940:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 11 150 theme glycosylation 2477:2489 arg1 process 2504:2510 a widely existing glycosylation modification process 2459:2510 a widely existing glycosylation modification process 2459:2510 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 0 151 theme stable 119:124 arg1 labeling 134:141 simultaneous stable isotope labeling 106:141 simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis 106:224 Quantitative O-glycomics based on improvement of the one-pot method for nonreductive O-glycan release and simultaneous stable isotope labeling with 1-(d0/d5)phenyl-3-methyl-5-pyrazolone followed by mass spectrometric analysis. 27585995 11 152 gly glycoproteins 2430:2442 arg1 glycoproteins 2430:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins 2397:2442 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 8 153 theme sample 1935:1940 arg1 approach 1955:1962 the corresponding sample purification approach 1917:1962 the one-pot reaction as well as the corresponding sample purification approach 1885:1962 We have refined the conditions for the one-pot reaction as well as the corresponding sample purification approach. 27585995 2 154 theme isotope 489:495 arg1 labeling 497:504 stable isotope labeling 482:504 stable isotope labeling followed by mass spectrometric analysis 482:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 6 155 theme complete 1492:1499 arg1 discrimination 1501:1514 complete discrimination 1492:1514 complete discrimination 1492:1514 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 1 156 theme O-glycans 305:313 arg1 analysis 280:287 quantitative analysis 267:287 quantitative analysis of glycoprotein O-glycans 267:313 Rapid, simple and versatile methods for quantitative analysis of glycoprotein O-glycans are urgently required for current studies on protein O-glycosylation patterns and the search for disease O-glycan biomarkers. 27585995 4 157 theme in 1103:1104 arg1 labeling 1124:1131 in situ chromophoric labeling 1103:1131 in situ chromophoric labeling of intact mucin-type O-glycans in one step 1103:1174 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 5 158 theme O-glycan 1253:1260 arg1 release 1262:1268 quantitative O-glycan release 1240:1268 quantitative O-glycan release 1240:1268 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 7 159 theme d0- 1635:1637 arg1 derivatives 1650:1660 the d0- and d5-PMP derivatives 1631:1660 derivatives 1650:1660 Moreover, the d0- and d5-PMP derivatives of O-glycans also have a relatively high hydrophobicity as well as a strong UV adsorption, especially suitable for high-resolution separation and high-sensitivity detection by RP-HPLC-UV. 27585995 14 160 gly glycoprotein 3118:3129 arg1 glycoprotein 3118:3129 glycoprotein O-glycans 3118:3139 glycoprotein O-glycans 3118:3139 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 2 161 theme mass 518:521 arg1 analysis 537:544 mass spectrometric analysis 518:544 mass spectrometric analysis 518:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 10 162 theme online 2287:2292 arg1 RP-HPLC-UV-ESI-MS/MS 2294:2313 online RP-HPLC-UV-ESI-MS/MS 2287:2313 online RP-HPLC-UV-ESI-MS/MS 2287:2313 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 5 163 with tagging 1274:1280 arg1 PMP 1367:1369 PMP 1367:1369 PMP 1367:1369 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 5 163 with tagging 1274:1280 arg1 1-phenyl-3-methyl-5-pyrazolone 1335:1364 either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone 1287:1364 1-phenyl-3-methyl-5-pyrazolone 1335:1364 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 5 163 with tagging 1274:1280 arg1 d0- 1310:1312 d0- 1310:1312 d0- 1310:1312 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 2 164 theme Relative 441:448 arg1 quantitation 450:461 Relative quantitation 441:461 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis 441:544 Relative quantitation of O-glycans using stable isotope labeling followed by mass spectrometric analysis represents an ideal and promising technique. 27585995 6 165 theme d5-PMP 1469:1474 arg1 forms 1476:1480 the d0- and d5-PMP forms 1457:1480 forms 1476:1480 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 13 166 gly glycoprotein 2977:2988 arg1 glycoprotein 2977:2988 various glycoprotein O-glycans 2969:2998 various glycoprotein O-glycans 2969:2998 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 3 167 theme small 713:717 arg1 difference 735:744 the too large or too small inconstant mass difference 692:744 the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry 618:924 However, it is hindered by the shortage of reliable nonreductive O-glycan release methods as well as the too large or too small inconstant mass difference between the light and heavy isotope form derivatives of O-glycans, which results in difficulties during the recognition and quantitative analysis of O-glycans by mass spectrometry. 27585995 5 168 with release 1262:1268 arg1 PMP 1367:1369 PMP 1367:1369 PMP 1367:1369 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 5 168 with release 1262:1268 arg1 1-phenyl-3-methyl-5-pyrazolone 1335:1364 either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone 1287:1364 1-phenyl-3-methyl-5-pyrazolone 1335:1364 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 5 168 with release 1262:1268 arg1 d0- 1310:1312 d0- 1310:1312 d0- 1310:1312 In this study, the one-pot method is optimized and applied for quantitative O-glycan release and tagging with either non-deuterated (d0-) or deuterated (d5-) 1-phenyl-3-methyl-5-pyrazolone (PMP). 27585995 14 169 from evaluation 3178:3187 arg1 level 3246:3250 expression level 3235:3250 expression level 3235:3250 The novel strategy introduced here provides a simple and versatile analytical method for the precise quantitation of glycoprotein O-glycans by mass spectrometry, enabling rapid evaluation of the differences or changes of O-glycans in expression level. 27585995 15 170 theme O-glycans 3415:3423 arg1 relationship 3399:3410 the complex structure-function relationship 3368:3410 the complex structure-function relationship of O-glycans 3368:3423 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 15 171 theme targets 3519:3525 arg1 search 3445:3450 the search 3441:3450 the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs 3441:3539 It is attractive for the field of quantitative/comparative O-glycomics, which has great significance for exploring the complex structure-function relationship of O-glycans, as well as for the search of O-glycan biomarkers of some major diseases and O-glycan related targets of some drugs. 27585995 4 172 theme mucin-type 1143:1152 arg1 O-glycans 1154:1162 intact mucin-type O-glycans 1136:1162 intact mucin-type O-glycans 1136:1162 Herein we report a facile and versatile O-glycan relative quantification strategy, based on an improved one-pot method that can quantitatively achieve nonreductive release and in situ chromophoric labeling of intact mucin-type O-glycans in one step. 27585995 6 173 theme labeled 1569:1575 arg1 O-glycans 1577:1585 these isotopically labeled O-glycans 1550:1585 these isotopically labeled O-glycans 1550:1585 The obtained O-glycan derivatives feature a permanent 10-Da mass difference between the d0- and d5-PMP forms, allowing complete discrimination and comparative quantification of these isotopically labeled O-glycans by mass spectrometric techniques. 27585995 13 174 from changes 2802:2808 arg1 level 2837:2841 expression level 2826:2841 expression level 2826:2841 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27585995 10 175 dep perch 2244:2248 arg1 eggs 2261:2264 eggs 2261:2264 eggs 2261:2264 Additionally, we have also successfully applied this method to the quantitative O-glycomic comparison between perch and salmon eggs by ESI-MS, MS/MS and online RP-HPLC-UV-ESI-MS/MS, demonstrating its excellent applicability to various complex biological samples. 27585995 11 176 theme modification 2491:2502 arg1 process 2504:2510 a widely existing glycosylation modification process 2459:2510 a widely existing glycosylation modification process 2459:2510 BIOLOGICAL SIGNIFICANCE O-Linked glycoproteins, generated via a widely existing glycosylation modification process on serine (Ser) or threonine (Thr) residues of nascent proteins, play essential roles in a series of biological processes. 27585995 13 177 theme many 2876:2879 arg1 diseases 2881:2888 many diseases 2876:2888 many diseases 2876:2888 Thus, the characteristic differences or changes of O-glycans in expression level usually relate to pathologies of many diseases and represent an important opportunity to uncover the functional mechanisms of various glycoprotein O-glycans. 27311011 7 0 theme structural 1220:1229 arg1 characterization 1231:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 5 1 theme single 918:923 arg1 subclass 925:932 a single subclass 916:932 a single subclass of IgG 916:939 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 1 2 theme significant 262:272 arg1 impact 274:279 a significant impact 260:279 a significant impact 260:279 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 2 3 theme glycosylation 372:384 arg1 sites 386:390 glycosylation sites 372:390 glycosylation sites 372:390 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 7 4 theme Such 1204:1207 arg1 characterization 1231:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 0 5 from Quantitation 0:11 arg1 Drugs 92:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 5 6 theme total 967:971 arg1 content 977:983 the total IgG content 963:983 the total IgG content 963:983 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 7 7 theme multilevel 1209:1218 arg1 characterization 1231:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization 1204:1246 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 6 8 theme occupancy 1178:1186 arg1 extent 1163:1168 the extent 1159:1168 the extent of site occupancy within minutes 1159:1201 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 5 9 theme IgG 973:975 arg1 content 977:983 the total IgG content 963:983 the total IgG content 963:983 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 7 10 theme therapeutic 1295:1305 arg1 antibodies 1307:1316 therapeutic antibodies 1295:1316 therapeutic antibodies 1295:1316 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 7 11 theme antibodies 1307:1316 arg1 development 1280:1290 the successful development 1265:1290 the successful development of therapeutic antibodies 1265:1316 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 6 12 theme MRM-based 1072:1080 arg1 method 1082:1087 an MRM-based method 1069:1087 an MRM-based method 1069:1087 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 3 13 theme specific 546:553 arg1 method 555:560 a sensitive and specific method 530:560 a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 530:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 2 14 theme glycan 425:430 arg1 changes 432:438 glycan changes 425:438 glycan changes specific to an individual protein within a complex mixture 425:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 2 15 theme complex 483:489 arg1 mixture 491:497 a complex mixture 481:497 a complex mixture 481:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 6 16 gly glycopeptides 1131:1143 arg2 glycopeptides 1131:1143 130 distinct glycopeptides 1118:1143 130 distinct glycopeptides 1118:1143 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 6 17 theme method 1082:1087 arg1 utility 1058:1064 the utility 1054:1064 the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes 1054:1201 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 1 18 theme particular 194:203 arg1 focus 205:209 particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions 194:314 particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions 194:314 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 4 19 theme IgG 786:788 arg1 subclass 790:797 the IgG subclass 782:797 the IgG subclass expressed in the drugs 782:820 Our results showed that irrespective of the IgG subclass expressed in the drugs, the N-glycopeptide profiles are nearly the same but differ in abundances. 27311011 5 20 theme rMAb 904:907 arg1 drugs 909:913 all rMAb drugs 900:913 all rMAb drugs 900:913 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 1 21 theme antibody 288:295 arg1 functions 306:314 the antibody effector functions 284:314 the antibody effector functions 284:314 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 2 22 theme in-depth 404:411 arg1 analysis 413:420 the in-depth analysis 400:420 the in-depth analysis of glycan changes specific to an individual protein within a complex mixture 400:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 0 23 theme Glycosylation 30:42 arg1 Quantitation 0:11 Quantitation 0:11 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 0:97 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 1 24 theme effector 297:304 arg1 functions 306:314 the antibody effector functions 284:314 the antibody effector functions 284:314 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 2 25 theme vast 321:324 arg1 heterogeneity 326:338 The vast heterogeneity 317:338 The vast heterogeneity of structures that exist across glycosylation sites 317:390 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 5 26 theme N-glycan 1005:1012 arg1 occupancy 1019:1027 97% N-glycan site occupancy 1001:1027 97% N-glycan site occupancy 1001:1027 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 0 27 theme Site-Specific 16:28 arg1 Glycosylation 30:42 Site-Specific Glycosylation 16:42 Site-Specific Glycosylation 16:42 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 5 28 theme content 977:983 arg1 content 977:983 the total IgG content 963:983 the total IgG content 963:983 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 5 28 theme content 977:983 arg1 % 958:958 97% 956:958 97% of the total IgG content 956:983 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 3 29 theme liquid 683:688 arg1 MS 723:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS 658:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 658:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 29 theme liquid 683:688 arg1 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 1 30 theme recombinant 125:135 arg1 rMAb 158:161 rMAb 158:161 rMAb 158:161 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 1 30 theme recombinant 125:135 arg1 antibody 148:155 recombinant monoclonal antibody 125:155 recombinant monoclonal antibody (rMAb) drugs 125:168 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 7 31 theme successful 1269:1278 arg1 development 1280:1290 the successful development 1265:1290 the successful development of therapeutic antibodies 1265:1316 Such multilevel structural characterization is important for the successful development of therapeutic antibodies. 27311011 0 32 theme Manufactured 47:58 arg1 Drugs 92:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 5 33 theme IgG 937:939 arg1 subclass 925:932 a single subclass 916:932 a single subclass of IgG 916:939 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 2 34 gly glycosylation 372:384 arg2 sites 386:390 glycosylation sites 372:390 glycosylation sites 372:390 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 3 35 theme chromatography-triple 690:710 arg1 MS 723:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS 658:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 658:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 35 theme chromatography-triple 690:710 arg1 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 36 theme reaction 629:636 arg1 MRM 650:652 MRM 650:652 MRM 650:652 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 36 theme reaction 629:636 arg1 monitoring 638:647 multiple reaction monitoring 620:647 multiple reaction monitoring (MRM) 620:653 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 2 37 theme specific 440:447 arg1 changes 432:438 glycan changes 425:438 glycan changes specific to an individual protein within a complex mixture 425:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 4 38 gly N-glycopeptide 827:840 arg2 N-glycopeptide 827:840 N-glycopeptide 827:840 N-glycopeptide 827:840 Our results showed that irrespective of the IgG subclass expressed in the drugs, the N-glycopeptide profiles are nearly the same but differ in abundances. 27311011 1 39 contain have 255:258 arg2 impact 274:279 a significant impact 260:279 a significant impact 260:279 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 1 39 contain have 255:258 arg1 changes 219:225 changes 219:225 changes in the attached glycans 219:249 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 1 40 theme attached 234:241 arg1 glycans 243:249 the attached glycans 230:249 the attached glycans 230:249 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 0 41 theme Monoclonal 72:81 arg1 Drugs 92:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 3 42 theme ultrahigh-performance 661:681 arg1 MS 723:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS 658:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 658:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 42 theme ultrahigh-performance 661:681 arg1 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 6 43 theme distinct 1122:1129 arg1 glycopeptides 1131:1143 130 distinct glycopeptides 1118:1143 130 distinct glycopeptides 1118:1143 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 1 44 theme monoclonal 137:146 arg1 rMAb 158:161 rMAb 158:161 rMAb 158:161 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 1 44 theme monoclonal 137:146 arg1 antibody 148:155 recombinant monoclonal antibody 125:155 recombinant monoclonal antibody (rMAb) drugs 125:168 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 0 45 theme Recombinant 60:70 arg1 Drugs 92:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 3 46 theme multiple 620:627 arg1 MRM 650:652 MRM 650:652 MRM 650:652 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 46 theme multiple 620:627 arg1 monitoring 638:647 multiple reaction monitoring 620:647 multiple reaction monitoring (MRM) 620:653 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 1 47 from changes 219:225 arg1 glycans 243:249 the attached glycans 230:249 the attached glycans 230:249 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 1 48 theme antibody 148:155 arg1 drugs 164:168 recombinant monoclonal antibody (rMAb) drugs 125:168 recombinant monoclonal antibody (rMAb) drugs 125:168 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 27311011 5 49 theme % 1003:1003 arg1 occupancy 1019:1027 97% N-glycan site occupancy 1001:1027 97% N-glycan site occupancy 1001:1027 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 6 50 theme site 1173:1176 arg1 occupancy 1178:1186 site occupancy 1173:1186 site occupancy 1173:1186 This study demonstrates the utility of an MRM-based method to rapidly characterize over 130 distinct glycopeptides and determine the extent of site occupancy within minutes. 27311011 4 51 theme N-glycopeptide 827:840 arg1 profiles 842:849 the N-glycopeptide profiles 823:849 the N-glycopeptide profiles 823:849 Our results showed that irrespective of the IgG subclass expressed in the drugs, the N-glycopeptide profiles are nearly the same but differ in abundances. 27311011 4 51 theme N-glycopeptide 827:840 arg1 same 866:869 same 866:869 same 866:869 Our results showed that irrespective of the IgG subclass expressed in the drugs, the N-glycopeptide profiles are nearly the same but differ in abundances. 27311011 5 52 theme site 1014:1017 arg1 occupancy 1019:1027 97% N-glycan site occupancy 1001:1027 97% N-glycan site occupancy 1001:1027 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 3 53 gly glycosylation 591:603 arg1 rMAbs 608:612 rMAbs 608:612 rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 608:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 54 theme site-specific 577:589 arg1 glycosylation 591:603 site-specific glycosylation 577:603 site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 577:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 55 theme quadrupole 712:721 arg1 MS 723:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS 658:724 an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 658:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 55 theme quadrupole 712:721 arg1 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 UHPLC-QqQ-MS 727:738 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 2 56 theme structures 343:352 arg1 heterogeneity 326:338 The vast heterogeneity 317:338 The vast heterogeneity of structures that exist across glycosylation sites 317:390 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 2 57 gly heterogeneity 326:338 arg1 structures 343:352 structures 343:352 structures that exist across glycosylation sites 343:390 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 2 58 theme changes 432:438 arg1 analysis 413:420 the in-depth analysis 400:420 the in-depth analysis of glycan changes specific to an individual protein within a complex mixture 400:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 5 59 theme 97 1001:1002 arg1 % 1003:1003 % 1003:1003 % 1003:1003 In all rMAb drugs, a single subclass of IgG comprised over 97% of the total IgG content and showed over 97% N-glycan site occupancy. 27311011 3 60 theme sensitive 532:540 arg1 method 555:560 a sensitive and specific method 530:560 a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 530:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 3 61 from glycosylation 591:603 arg1 rMAbs 608:612 rMAbs 608:612 rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS) 608:739 In this study, we established a sensitive and specific method for monitoring site-specific glycosylation in rMAbs using multiple reaction monitoring (MRM) on an ultrahigh-performance liquid chromatography-triple quadrupole MS (UHPLC-QqQ-MS). 27311011 0 62 theme Antibody 83:90 arg1 Drugs 92:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Manufactured Recombinant Monoclonal Antibody Drugs 47:96 Quantitation of Site-Specific Glycosylation in Manufactured Recombinant Monoclonal Antibody Drugs. 27311011 2 63 theme individual 455:464 arg1 protein 466:472 an individual protein 452:472 an individual protein within a complex mixture 452:497 The vast heterogeneity of structures that exist across glycosylation sites hinders the in-depth analysis of glycan changes specific to an individual protein within a complex mixture. 27311011 1 64 theme drugs 164:168 arg1 development 110:120 the development 106:120 the development of recombinant monoclonal antibody (rMAb) drugs 106:168 During the development of recombinant monoclonal antibody (rMAb) drugs, glycosylation receives particular focus because changes in the attached glycans can have a significant impact on the antibody effector functions. 26863921 0 0 theme archaeon 103:110 arg1 volcanii 122:129 the halophilic archaeon Haloferax volcanii 88:129 the halophilic archaeon Haloferax volcanii 88:129 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 3 1 theme mass 448:451 arg1 MS 467:468 MS 467:468 MS 467:468 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 1 theme mass 448:451 arg1 spectrometry 453:464 mass spectrometry 448:464 mass spectrometry (MS) 448:469 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 0 2 theme halophilic 92:101 arg1 volcanii 122:129 the halophilic archaeon Haloferax volcanii 88:129 the halophilic archaeon Haloferax volcanii 88:129 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 2 3 theme Agl 387:389 arg1 proteins 416:423 Agl (archaeal glycosylation) proteins 387:423 Agl (archaeal glycosylation) proteins 387:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 5 4 theme methylated 1328:1337 arg1 GlcA 1339:1342 a methylated GlcA 1326:1342 a methylated GlcA 1326:1342 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 0 5 theme volcanii 122:129 arg1 glycoproteins 71:83 glycoproteins 71:83 glycoproteins of the halophilic archaeon Haloferax volcanii 71:129 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 5 6 theme hexuronic 1275:1283 arg1 acid 1285:1288 a hexuronic acid 1273:1288 a hexuronic acid 1273:1288 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 7 theme present 1173:1179 arg1 analysis 1181:1188 The present analysis 1169:1188 The present analysis 1169:1188 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 3 8 theme glycan 557:562 arg1 structure 539:547 the structure 535:547 the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279 535:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 0 9 theme Haloferax 112:120 arg1 volcanii 122:129 the halophilic archaeon Haloferax volcanii 88:129 the halophilic archaeon Haloferax volcanii 88:129 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 5 10 theme sugar 1246:1250 arg1 ester 1264:1268 a methyl ester 1255:1268 a methyl ester of a hexuronic acid 1255:1288 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 10 theme sugar 1246:1250 arg1 assignment 1216:1225 the earlier assignment 1204:1225 the earlier assignment of the penultimate sugar 1204:1250 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 3 11 gly glycoprotein 620:631 arg1 glycoprotein 620:631 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 2 12 mod modified 322:329 arg1 glycoprotein 305:316 the S-layer glycoprotein 293:316 the S-layer glycoprotein 293:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 12 mod modified 322:329 arg1 glycoproteins 271:283 glycoproteins 271:283 glycoproteins such as the S-layer glycoprotein 271:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 12 mod modified 322:329 arg3 pentasaccharide 346:360 an N-linked pentasaccharide 334:360 an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins 334:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 4 13 theme pentasaccharide 1138:1152 arg1 end 1127:1129 the non-reducing end 1110:1129 the non-reducing end of the pentasaccharide to be mannose 1110:1166 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 2 14 gly glycoprotein 305:316 arg1 glycoprotein 305:316 the S-layer glycoprotein 293:316 the S-layer glycoprotein 293:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 1 15 theme life 219:222 arg1 life 219:222 life 219:222 life 219:222 N-Glycosylation is a post-translational modification performed in all three domains of life. 26863921 1 15 theme life 219:222 arg1 domains 208:214 all three domains 198:214 all three domains of life 198:222 N-Glycosylation is a post-translational modification performed in all three domains of life. 26863921 7 16 theme N-glycosylation 1442:1456 arg1 pathway 1458:1464 volcanii N-glycosylation pathway 1433:1464 volcanii N-glycosylation pathway from previous genetic and biochemical 1433:1502 volcanii N-glycosylation pathway from previous genetic and biochemical efforts while providing new insight into the process. 26863921 6 17 theme good 1378:1381 arg1 agreement 1383:1391 good agreement 1378:1391 good agreement with what was already known of the Hfx 1378:1430 The assignments made here are in good agreement with what was already known of the Hfx. 26863921 7 18 from genetic 1480:1486 arg1 pathway 1458:1464 volcanii N-glycosylation pathway 1433:1464 volcanii N-glycosylation pathway from previous genetic and biochemical 1433:1502 volcanii N-glycosylation pathway from previous genetic and biochemical efforts while providing new insight into the process. 26863921 3 19 gly N-glycosylation 633:647 arg2 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 19 gly N-glycosylation 633:647 arg2 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 19 gly N-glycosylation 633:647 arg2 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 19 gly N-glycosylation 633:647 arg2 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 19 gly N-glycosylation 633:647 arg2 seven 597:601 seven 597:601 seven 597:601 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 19 gly N-glycosylation 633:647 arg2 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 5 20 theme methyl 1257:1262 arg1 assignment 1216:1225 the earlier assignment 1204:1225 the earlier assignment of the penultimate sugar 1204:1250 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 20 theme methyl 1257:1262 arg1 ester 1264:1268 a methyl ester 1255:1268 a methyl ester of a hexuronic acid 1255:1288 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 4 21 theme glucuronic 758:767 arg1 acid 769:772 glucuronic acid 758:772 glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn 758:809 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 4 21 theme glucuronic 758:767 arg1 GlcA 775:778 GlcA 775:778 GlcA 775:778 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 2 22 theme proteins 416:423 arg1 series 377:382 a series 375:382 a series of Agl (archaeal glycosylation) proteins 375:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 0 23 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of the N-linked pentasaccharide 0:58 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 3 24 theme N-glycosylation 633:647 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 24 theme N-glycosylation 633:647 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 24 theme N-glycosylation 633:647 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 24 theme N-glycosylation 633:647 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 24 theme N-glycosylation 633:647 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 four 585:588 four 585:588 four 585:588 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg2 glycan 557:562 this glycan 552:562 this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279 552:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 25 attach attached 564:571 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 4 26 theme Such 700:703 arg1 approaches 705:714 Such approaches 700:714 Such approaches 700:714 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 4 27 theme radiolabeling 1062:1074 arg1 experiments 1076:1086 previous MS and radiolabeling experiments 1046:1086 experiments 1076:1086 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 0 28 link N-linked 35:42 arg1 pentasaccharide 44:58 the N-linked pentasaccharide 31:58 the N-linked pentasaccharide 31:58 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 2 29 theme N-linked 337:344 arg1 pentasaccharide 346:360 an N-linked pentasaccharide 334:360 an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins 334:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 3 30 theme nuclear 475:481 arg1 resonance 492:500 nuclear magnetic resonance 475:500 nuclear magnetic resonance spectroscopy 475:513 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 4 31 theme acid 769:772 arg1 -β1,4-GlcA-β1,4-glucose-β1-Asn 780:809 glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn 758:809 glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn 758:809 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 3 32 theme magnetic 483:490 arg1 resonance 492:500 nuclear magnetic resonance 475:500 nuclear magnetic resonance spectroscopy 475:513 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 0 33 theme pentasaccharide 44:58 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of the N-linked pentasaccharide 0:58 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 7 34 theme volcanii 1433:1440 arg1 pathway 1458:1464 volcanii N-glycosylation pathway 1433:1464 volcanii N-glycosylation pathway from previous genetic and biochemical 1433:1502 volcanii N-glycosylation pathway from previous genetic and biochemical efforts while providing new insight into the process. 26863921 0 35 theme N-linked 35:42 arg1 pentasaccharide 44:58 the N-linked pentasaccharide 31:58 the N-linked pentasaccharide 31:58 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 2 36 link N-linked 337:344 arg1 pentasaccharide 346:360 an N-linked pentasaccharide 334:360 an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins 334:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 37 gly glycoproteins 271:283 arg1 glycoprotein 305:316 the S-layer glycoprotein 293:316 the S-layer glycoprotein 293:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 37 gly glycoproteins 271:283 arg1 glycoproteins 271:283 glycoproteins 271:283 glycoproteins such as the S-layer glycoprotein 271:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 3 38 theme putative 603:610 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 38 theme putative 603:610 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 38 theme putative 603:610 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 38 theme putative 603:610 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 38 theme putative 603:610 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 5 39 theme acid 1285:1288 arg1 assignment 1216:1225 the earlier assignment 1204:1225 the earlier assignment of the penultimate sugar 1204:1250 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 39 theme acid 1285:1288 arg1 ester 1264:1268 a methyl ester 1255:1268 a methyl ester of a hexuronic acid 1255:1288 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 40 theme penultimate 1234:1244 arg1 sugar 1246:1250 the penultimate sugar 1230:1250 the penultimate sugar 1230:1250 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 3 41 used used 520:523 arg2 MS 467:468 MS 467:468 MS 467:468 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 41 used used 520:523 arg2 spectroscopy 502:513 nuclear magnetic resonance spectroscopy 475:513 nuclear magnetic resonance spectroscopy 475:513 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 41 used used 520:523 arg2 spectrometry 453:464 mass spectrometry 448:464 mass spectrometry (MS) 448:469 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 42 theme present 433:439 arg1 study 441:445 the present study 429:445 the present study 429:445 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 2 43 theme archaea 243:249 arg1 volcanii 261:268 the halophilic archaea Haloferax volcanii 228:268 the halophilic archaea Haloferax volcanii 228:268 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 44 theme Haloferax 251:259 arg1 volcanii 261:268 the halophilic archaea Haloferax volcanii 228:268 the halophilic archaea Haloferax volcanii 228:268 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 3 45 theme glycoprotein 620:631 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 45 theme glycoprotein 620:631 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 45 theme glycoprotein 620:631 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 45 theme glycoprotein 620:631 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 45 theme glycoprotein 620:631 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 2 46 theme halophilic 232:241 arg1 volcanii 261:268 the halophilic archaea Haloferax volcanii 228:268 the halophilic archaea Haloferax volcanii 228:268 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 2 47 theme S-layer 297:303 arg1 glycoprotein 305:316 the S-layer glycoprotein 293:316 the S-layer glycoprotein 293:316 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 4 48 theme methyl-O-4-GlcA-β-1,4-galacturonic 847:880 arg1 acid-α1,4-GlcA-β1,4-glucose-β1-Asn 882:915 methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn 847:915 methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn 847:915 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 1 49 theme post-translational 153:170 arg1 modification 172:183 a post-translational modification 151:183 a post-translational modification performed in all three domains of life 151:222 N-Glycosylation is a post-translational modification performed in all three domains of life. 26863921 1 49 theme post-translational 153:170 arg1 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation is a post-translational modification performed in all three domains of life. 26863921 7 50 theme new 1528:1530 arg1 insight 1532:1538 new insight 1528:1538 new insight into the process 1528:1555 volcanii N-glycosylation pathway from previous genetic and biochemical efforts while providing new insight into the process. 26863921 4 51 theme previous 1046:1053 arg1 MS 1055:1056 previous MS and radiolabeling experiments 1046:1086 MS 1055:1056 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 3 52 theme sites 649:653 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 52 theme sites 649:653 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 52 theme sites 649:653 arg1 four 585:588 four 585:588 four 585:588 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 52 theme sites 649:653 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 52 theme sites 649:653 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 52 theme sites 649:653 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 7 53 from biochemical 1492:1502 arg1 pathway 1458:1464 volcanii N-glycosylation pathway 1433:1464 volcanii N-glycosylation pathway from previous genetic and biochemical 1433:1502 volcanii N-glycosylation pathway from previous genetic and biochemical efforts while providing new insight into the process. 26863921 4 54 theme GlcA-β-1,4-galacturonic 981:1003 arg1 acid-α1,4-GlcA-β1,4-glucose-β1-Asn 1005:1038 hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn 957:1038 hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn 957:1038 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 3 55 theme resonance 492:500 arg1 spectroscopy 502:513 nuclear magnetic resonance spectroscopy 475:513 nuclear magnetic resonance spectroscopy 475:513 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 5 56 theme earlier 1208:1214 arg1 ester 1264:1268 a methyl ester 1255:1268 a methyl ester of a hexuronic acid 1255:1288 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 5 56 theme earlier 1208:1214 arg1 assignment 1216:1225 the earlier assignment 1204:1225 the earlier assignment of the penultimate sugar 1204:1250 The present analysis thus corrects the earlier assignment of the penultimate sugar as a methyl ester of a hexuronic acid, instead revealing this sugar to be a methylated GlcA. 26863921 3 57 theme S-layer 612:618 arg1 Asn-13 663:668 Asn-13 663:668 Asn-13 663:668 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 57 theme S-layer 612:618 arg1 Asn-279 691:697 Asn-279 691:697 Asn-279 691:697 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 57 theme S-layer 612:618 arg1 Asn-274 679:685 Asn-274 679:685 Asn-274 679:685 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 57 theme S-layer 612:618 arg1 Asn-83 671:676 Asn-83 671:676 Asn-83 671:676 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 3 57 theme S-layer 612:618 arg1 sites 649:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 the seven putative S-layer glycoprotein N-glycosylation sites 593:653 In the present study, mass spectrometry (MS) and nuclear magnetic resonance spectroscopy were used to define the structure of this glycan attached to at least four of the seven putative S-layer glycoprotein N-glycosylation sites, namely Asn-13, Asn-83, Asn-274 and Asn-279. 26863921 2 58 theme glycosylation 401:413 arg1 proteins 416:423 Agl (archaeal glycosylation) proteins 387:423 Agl (archaeal glycosylation) proteins 387:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 26863921 4 59 theme non-reducing 1114:1125 arg1 end 1127:1129 the non-reducing end 1110:1129 the non-reducing end of the pentasaccharide to be mannose 1110:1166 Such approaches detected a trisaccharide corresponding to glucuronic acid (GlcA)-β1,4-GlcA-β1,4-glucose-β1-Asn, a tetrasaccharide corresponding to methyl-O-4-GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, and a pentasaccharide corresponding to hexose-1,2-[methyl-O-4-]GlcA-β-1,4-galacturonic acid-α1,4-GlcA-β1,4-glucose-β1-Asn, with previous MS and radiolabeling experiments showing the hexose at the non-reducing end of the pentasaccharide to be mannose. 26863921 0 60 gly glycoproteins 71:83 arg1 glycoproteins 71:83 glycoproteins 71:83 glycoproteins of the halophilic archaeon Haloferax volcanii 71:129 Structural characterization of the N-linked pentasaccharide decorating glycoproteins of the halophilic archaeon Haloferax volcanii. 26863921 2 61 theme archaeal 392:399 arg1 proteins 416:423 Agl (archaeal glycosylation) proteins 387:423 Agl (archaeal glycosylation) proteins 387:423 In the halophilic archaea Haloferax volcanii, glycoproteins such as the S-layer glycoprotein are modified by an N-linked pentasaccharide assembled by a series of Agl (archaeal glycosylation) proteins. 27616296 3 0 theme liquid 906:911 arg1 chromatography 913:926 online hydrophilic interaction liquid chromatography 875:926 online hydrophilic interaction liquid chromatography 875:926 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 1 theme mass 844:847 arg1 ESI-MS 863:868 ESI-MS 863:868 ESI-MS 863:868 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 1 theme mass 844:847 arg1 spectrometry 849:860 electrospray ionization mass spectrometry 820:860 electrospray ionization mass spectrometry (ESI-MS) 820:869 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 2 theme moieties 559:566 arg1 features 536:543 structural features 525:543 structural features of the glycan moieties of more soybean glycoproteins 525:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 3 3 theme in-gel 697:702 arg1 release 713:719 in-gel N-glycan release 697:719 in-gel N-glycan release 697:719 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 4 with related 166:172 arg1 allergies 184:192 food allergies 179:192 food allergies 179:192 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 3 5 theme online 875:880 arg1 chromatography 913:926 online hydrophilic interaction liquid chromatography 875:926 online hydrophilic interaction liquid chromatography 875:926 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 6 theme glycoallergens 262:275 arg1 screening 199:207 screening 199:207 screening 199:207 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 1 6 theme glycoallergens 262:275 arg1 analysis 224:231 structural analysis 213:231 structural analysis 213:231 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 1 7 gly Glycosylation 103:115 arg1 proteins 125:132 many proteins 120:132 many proteins 120:132 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 3 8 theme N-glycan 704:711 arg1 release 713:719 in-gel N-glycan release 697:719 in-gel N-glycan release 697:719 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 4 9 dep two 1088:1090 arg1 contain 1099:1105 contain 1099:1105 mainly contain core α1,3-fucosylated glycans 1092:1135 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 3 10 theme hydrophilic 882:892 arg1 chromatography 913:926 online hydrophilic interaction liquid chromatography 875:926 online hydrophilic interaction liquid chromatography 875:926 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 11 theme electrospray 820:831 arg1 ESI-MS 863:868 ESI-MS 863:868 ESI-MS 863:868 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 11 theme electrospray 820:831 arg1 spectrometry 849:860 electrospray ionization mass spectrometry 820:860 electrospray ionization mass spectrometry (ESI-MS) 820:869 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 12 gly glycoproteins 244:256 arg1 glycoproteins 244:256 related glycoproteins 236:256 related glycoproteins 236:256 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 3 13 theme ionization 954:963 arg1 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 13 theme ionization 954:963 arg1 spectrometry 977:988 electrospray ionization tandem mass spectrometry 941:988 electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 941:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 14 theme ionization 833:842 arg1 ESI-MS 863:868 ESI-MS 863:868 ESI-MS 863:868 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 14 theme ionization 833:842 arg1 spectrometry 849:860 electrospray ionization mass spectrometry 820:860 electrospray ionization mass spectrometry (ESI-MS) 820:869 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 15 theme structural 525:534 arg1 features 536:543 structural features 525:543 structural features of the glycan moieties of more soybean glycoproteins 525:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 16 theme N-glycoform 347:357 arg1 analysis 359:366 detailed N-glycoform analysis 338:366 detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 338:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 3 17 theme electrospray 941:952 arg1 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 17 theme electrospray 941:952 arg1 spectrometry 977:988 electrospray ionization tandem mass spectrometry 941:988 electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 941:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 18 theme protein 409:415 arg1 SPI 426:428 SPI 426:428 SPI 426:428 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 18 theme protein 409:415 arg1 isolate 417:423 soybean protein isolate 401:423 soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 401:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 19 theme sulfate 459:465 arg1 SDS-PAGE 503:510 SDS-PAGE 503:510 SDS-PAGE 503:510 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 19 theme sulfate 459:465 arg1 electrophoresis 486:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis 444:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 444:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 20 theme sodium 444:449 arg1 SDS-PAGE 503:510 SDS-PAGE 503:510 SDS-PAGE 503:510 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 20 theme sodium 444:449 arg1 electrophoresis 486:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis 444:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 444:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 1 21 theme food 179:182 arg1 allergies 184:192 food allergies 179:192 food allergies 179:192 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 0 22 theme Spectrometric 5:17 arg1 Analysis 19:26 Mass Spectrometric Analysis 0:26 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins 0:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 6 23 theme allergens 1482:1490 arg1 research 1462:1469 research 1462:1469 research of soybean allergens 1462:1490 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 3 24 theme one-pot 742:748 arg1 method 750:755 a one-pot method 740:755 a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 740:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 0 25 theme Mass 0:3 arg1 Analysis 19:26 Mass Spectrometric Analysis 0:26 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins 0:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 1 26 from studies 295:301 arg1 field 311:315 this field 306:315 this field 306:315 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 6 27 theme structural 1437:1446 arg1 reference 1448:1456 significant structural reference 1425:1456 significant structural reference for research of soybean allergens 1425:1490 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 3 28 theme mass 972:975 arg1 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 28 theme mass 972:975 arg1 spectrometry 977:988 electrospray ionization tandem mass spectrometry 941:988 electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 941:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 29 theme fractions 388:396 arg1 analysis 359:366 detailed N-glycoform analysis 338:366 detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 338:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 4 30 contain have 1146:1149 arg1 six 1142:1144 six 1142:1144 six 1142:1144 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 30 contain have 1146:1149 arg1 two 1088:1090 two 1088:1090 two 1088:1090 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 30 contain have 1146:1149 arg2 glycans 1154:1160 no glycans 1151:1160 no glycans 1151:1160 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 31 contain containing 1050:1059 arg2 glycans 1079:1085 oligomannose-type glycans 1061:1085 oligomannose-type glycans 1061:1085 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 31 contain containing 1050:1059 arg1 bands 1037:1041 seven bands 1031:1041 seven bands mainly containing oligomannose-type glycans 1031:1085 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 31 contain containing 1050:1059 arg1 result 1014:1019 a result 1012:1019 a result 1012:1019 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 4 32 theme oligomannose-type 1061:1077 arg1 glycans 1079:1085 oligomannose-type glycans 1061:1085 oligomannose-type glycans 1061:1085 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 5 33 theme core 1209:1212 arg1 glycoproteins 1300:1312 glycoproteins 1300:1312 glycoproteins 1300:1312 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 33 theme core 1209:1212 arg1 N-glycans 1231:1239 core α1,3-fucosylated N-glycans 1209:1239 core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1209:1295 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 33 theme core 1209:1212 arg1 N-glycoforms 1324:1335 their N-glycoforms 1318:1335 their N-glycoforms 1318:1335 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 2 34 gly glycoprotein 375:386 arg1 glycoprotein 375:386 all glycoprotein fractions 371:396 all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 371:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 6 35 theme soybean 1383:1389 arg1 glycosylation 1399:1411 soybean protein glycosylation 1383:1411 soybean protein glycosylation 1383:1411 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 1 36 theme many 120:123 arg1 proteins 125:132 many proteins 120:132 many proteins 120:132 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 2 37 theme polyacrylamide 467:480 arg1 SDS-PAGE 503:510 SDS-PAGE 503:510 SDS-PAGE 503:510 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 37 theme polyacrylamide 467:480 arg1 electrophoresis 486:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis 444:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 444:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 6 38 theme protein 1391:1397 arg1 glycosylation 1399:1411 soybean protein glycosylation 1383:1411 soybean protein glycosylation 1383:1411 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 5 39 theme α1,3-fucosylated 1214:1229 arg1 glycoproteins 1300:1312 glycoproteins 1300:1312 glycoproteins 1300:1312 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 39 theme α1,3-fucosylated 1214:1229 arg1 N-glycans 1231:1239 core α1,3-fucosylated N-glycans 1209:1239 core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1209:1295 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 39 theme α1,3-fucosylated 1214:1229 arg1 N-glycoforms 1324:1335 their N-glycoforms 1318:1335 their N-glycoforms 1318:1335 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 2 40 theme glycan 552:557 arg1 moieties 559:566 the glycan moieties 548:566 the glycan moieties of more soybean glycoproteins 548:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 5 41 theme discloses 1266:1274 arg1 bands 1276:1280 discloses bands 1266:1280 discloses bands 1266:1280 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 6 42 theme significant 1425:1435 arg1 reference 1448:1456 significant structural reference 1425:1456 significant structural reference for research of soybean allergens 1425:1490 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 3 43 theme tandem 965:970 arg1 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 HILIC-ESI-MS/MS 991:1005 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 3 43 theme tandem 965:970 arg1 spectrometry 977:988 electrospray ionization tandem mass spectrometry 941:988 electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 941:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 44 gly glycoproteins 584:596 arg1 glycoproteins 584:596 more soybean glycoproteins 571:596 more soybean glycoproteins 571:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 0 45 theme N-Glycoforms 31:42 arg1 Analysis 19:26 Mass Spectrometric Analysis 0:26 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins 0:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 2 46 theme gel 482:484 arg1 SDS-PAGE 503:510 SDS-PAGE 503:510 SDS-PAGE 503:510 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 46 theme gel 482:484 arg1 electrophoresis 486:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis 444:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 444:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 3 47 theme generated 641:649 arg1 bands 659:663 the generated protein bands 637:663 the generated protein bands 637:663 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 48 theme structural 213:222 arg1 analysis 224:231 structural analysis 213:231 structural analysis 213:231 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 0 49 theme Allergenic 55:64 arg1 Glycoproteins 66:78 Soybean Allergenic Glycoproteins 47:78 Soybean Allergenic Glycoproteins 47:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 3 50 theme protein 651:657 arg1 bands 659:663 the generated protein bands 637:663 the generated protein bands 637:663 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 51 theme proteins 125:132 arg1 Glycosylation 103:115 Glycosylation 103:115 Glycosylation of many proteins 103:132 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 0 52 theme Soybean 47:53 arg1 Glycoproteins 66:78 Soybean Allergenic Glycoproteins 47:78 Soybean Allergenic Glycoproteins 47:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 5 53 theme first 1181:1185 arg1 report 1187:1192 the first report 1177:1192 the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms 1177:1335 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 53 theme first 1181:1185 arg1 study 1168:1172 This study 1163:1172 This study 1163:1172 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 2 54 theme glycoproteins 584:596 arg1 moieties 559:566 the glycan moieties 548:566 the glycan moieties of more soybean glycoproteins 548:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 5 55 gly α1,3-fucosylated 1214:1229 arg1 glycoproteins 1300:1312 glycoproteins 1300:1312 glycoproteins 1300:1312 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 55 gly α1,3-fucosylated 1214:1229 arg1 N-glycans 1231:1239 core α1,3-fucosylated N-glycans 1209:1239 core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1209:1295 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 55 gly α1,3-fucosylated 1214:1229 arg1 N-glycoforms 1324:1335 their N-glycoforms 1318:1335 their N-glycoforms 1318:1335 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 2 56 theme detailed 338:345 arg1 analysis 359:366 detailed N-glycoform analysis 338:366 detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 338:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 4 57 theme α1,3-fucosylated 1112:1127 arg1 glycans 1129:1135 core α1,3-fucosylated glycans 1107:1135 core α1,3-fucosylated glycans 1107:1135 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 2 58 theme soybean 576:582 arg1 glycoproteins 584:596 more soybean glycoproteins 571:596 more soybean glycoproteins 571:596 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 4 59 theme core 1107:1110 arg1 glycans 1129:1135 core α1,3-fucosylated glycans 1107:1135 core α1,3-fucosylated glycans 1107:1135 As a result, we found seven bands mainly containing oligomannose-type glycans; two mainly contain core α1,3-fucosylated glycans, and six have no glycans. 27616296 5 60 from N-glycans 1231:1239 arg1 bands 1244:1248 bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1244:1295 bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1244:1295 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 6 61 theme soybean 1474:1480 arg1 allergens 1482:1490 soybean allergens 1474:1490 soybean allergens 1474:1490 Therefore, it can expand our knowledge about soybean protein glycosylation and provide significant structural reference for research of soybean allergens. 27616296 2 62 theme isolate 417:423 arg1 fractions 388:396 all glycoprotein fractions 371:396 all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 371:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 63 theme glycoprotein 375:386 arg1 fractions 388:396 all glycoprotein fractions 371:396 all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 371:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 64 theme soybean 401:407 arg1 SPI 426:428 SPI 426:428 SPI 426:428 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 64 theme soybean 401:407 arg1 isolate 417:423 soybean protein isolate 401:423 soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 401:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 1 65 theme related 236:242 arg1 glycoproteins 244:256 related glycoproteins 236:256 related glycoproteins 236:256 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 0 66 theme Glycoproteins 66:78 arg1 N-Glycoforms 31:42 N-Glycoforms 31:42 N-Glycoforms of Soybean Allergenic Glycoproteins 31:78 Mass Spectrometric Analysis of N-Glycoforms of Soybean Allergenic Glycoproteins Separated by SDS-PAGE. 27616296 5 67 gly glycoproteins 1300:1312 arg1 glycoproteins 1300:1312 glycoproteins 1300:1312 glycoproteins 1300:1312 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 5 67 gly glycoproteins 1300:1312 arg1 N-glycans 1231:1239 core α1,3-fucosylated N-glycans 1209:1239 core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 1209:1295 This study is the first report that discovers core α1,3-fucosylated N-glycans in bands 1, 2, and 6 and discloses bands 3, 4, 5, and 7 as glycoproteins and their N-glycoforms. 27616296 3 68 theme detailed 799:806 arg1 analysis 808:815 detailed analysis 799:815 detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS) 799:1006 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 2 69 theme dodecyl 451:457 arg1 SDS-PAGE 503:510 SDS-PAGE 503:510 SDS-PAGE 503:510 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 2 69 theme dodecyl 451:457 arg1 electrophoresis 486:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis 444:500 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 444:511 Herein, we describe detailed N-glycoform analysis of all glycoprotein fractions of soybean protein isolate (SPI) separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to disclose structural features of the glycan moieties of more soybean glycoproteins. 27616296 3 70 theme interaction 894:904 arg1 chromatography 913:926 online hydrophilic interaction liquid chromatography 875:926 online hydrophilic interaction liquid chromatography 875:926 SPI was fractionated by SDS-PAGE, and the generated protein bands were recovered and subjected to in-gel N-glycan release and labeling using a one-pot method newly developed by our group, followed by detailed analysis by electrospray ionization mass spectrometry (ESI-MS) and online hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). 27616296 1 71 theme glycoproteins 244:256 arg1 screening 199:207 screening 199:207 screening 199:207 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 27616296 1 71 theme glycoproteins 244:256 arg1 analysis 224:231 structural analysis 213:231 structural analysis 213:231 Glycosylation of many proteins has been revealed to be closely related with food allergies, and screening and structural analysis of related glycoproteins and glycoallergens are essential for studies in this field. 25940448 10 0 theme genes 1630:1634 arg1 prediction 1607:1616 the functional prediction 1592:1616 the functional prediction of novel GR genes 1592:1634 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 7 1 from glycans 1220:1226 arg1 Notch 1209:1213 Notch 1209:1213 Notch 1209:1213 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 10 2 theme Phenotypic 1540:1549 arg1 clustering 1551:1560 Phenotypic clustering 1540:1560 Phenotypic clustering 1540:1560 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 10 3 theme novel 1621:1625 arg1 genes 1630:1634 novel GR genes 1621:1634 novel GR genes 1621:1634 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 6 4 theme thick 1079:1083 arg1 veins 1085:1089 thick veins 1079:1089 thick veins 1079:1089 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 5 5 theme Drosophila 803:812 arg1 125 834:836 125 834:836 125 834:836 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 5 theme Drosophila 803:812 arg1 genes 827:831 Drosophila GR candidate genes 803:831 Drosophila GR candidate genes (125) 803:837 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 7 6 theme GAGs 1176:1179 arg1 pathways 1164:1171 the biosynthetic pathways 1147:1171 the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans) 1147:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 7 dep ends 1258:1261 arg1 termed 1271:1276 termed 1271:1276 termed terminal domains of glycans 1271:1304 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 8 theme glycan 1199:1204 arg1 pathways 1164:1171 the biosynthetic pathways 1147:1171 the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans) 1147:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 8 9 theme domains 1406:1412 arg1 formation 1384:1392 formation 1384:1392 formation of terminal domains 1384:1412 Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. 25940448 7 10 theme terminal 1278:1285 arg1 glycans 1298:1304 glycans 1298:1304 glycans 1298:1304 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 10 theme terminal 1278:1285 arg1 domains 1287:1293 terminal domains 1278:1293 terminal domains of glycans 1278:1304 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 5 11 link O-linked 916:923 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 11 link O-linked 916:923 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 2 12 theme core 314:317 arg1 region 319:324 the common core region 303:324 the common core region of glycosaminoglycans (GAGs) 303:353 For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. 25940448 6 13 theme extra 1066:1070 arg1 veins 1072:1076 extra veins 1066:1076 extra veins 1066:1076 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 6 14 theme regulated 979:987 arg1 silencing 989:997 Spatiotemporally regulated silencing 962:997 Spatiotemporally regulated silencing 962:997 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 8 15 theme phenotypic 1321:1330 arg1 clustering 1332:1341 the phenotypic clustering 1317:1341 the phenotypic clustering 1317:1341 Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. 25940448 1 16 theme Glycan 91:96 arg1 structures 98:107 Glycan structures 91:107 Glycan structures 91:107 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 5 17 theme unknown 945:951 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 17 theme unknown 945:951 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 4 18 theme proteoglycan 747:758 arg1 proteins 765:772 proteoglycan core proteins 747:772 proteoglycan core proteins 747:772 To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. 25940448 7 19 theme glycans 1220:1226 arg1 pathways 1164:1171 the biosynthetic pathways 1147:1171 the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans) 1147:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 20 theme Fringe-dependent 1182:1197 arg1 glycan 1199:1204 Fringe-dependent glycan 1182:1204 Fringe-dependent glycan on Notch 1182:1213 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 5 21 theme GAGs 900:903 arg1 synthesis 887:895 synthesis 887:895 synthesis of GAGs 887:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 7 22 from pathways 1164:1171 arg1 Notch 1209:1213 Notch 1209:1213 Notch 1209:1213 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 23 from Notch 1209:1213 arg1 pathways 1164:1171 the biosynthetic pathways 1147:1171 the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans) 1147:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 24 theme nonreducing 1246:1256 arg1 ends 1258:1261 nonreducing ends 1246:1261 nonreducing ends (herein termed terminal domains of glycans) 1246:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 10 25 theme GR 1627:1628 arg1 genes 1630:1634 novel GR genes 1621:1634 novel GR genes 1621:1634 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 9 26 link N-linked 1522:1529 arg1 glycans 1531:1537 terminal N-linked glycans 1513:1537 terminal N-linked glycans 1513:1537 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 3 27 theme phenotypic 645:654 arg1 abnormality 656:666 the same phenotypic abnormality 636:666 the same phenotypic abnormality 636:666 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 3 28 theme glycan 613:618 arg1 synthesis 591:599 synthesis 591:599 synthesis of the same glycan 591:618 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 4 29 theme proteins 765:772 arg1 silencing 712:720 comprehensive silencing 698:720 comprehensive silencing of genes encoding GR and proteoglycan core proteins 698:772 To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. 25940448 0 30 theme Phenotype-based 0:14 arg1 clustering 16:25 Phenotype-based clustering 0:25 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 0:89 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 25940448 6 31 dep types 1059:1063 arg1 veins 1072:1076 extra veins 1066:1076 extra veins 1066:1076 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 6 31 dep types 1059:1063 arg1 veins 1085:1089 thick veins 1079:1089 thick veins 1079:1089 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 6 31 dep types 1059:1063 arg1 depigmentation 1096:1109 depigmentation 1096:1109 depigmentation 1096:1109 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 4 32 theme core 760:763 arg1 proteins 765:772 proteoglycan core proteins 747:772 proteoglycan core proteins 747:772 To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. 25940448 0 33 theme glycosylation-related 30:50 arg1 genes 52:56 glycosylation-related genes 30:56 glycosylation-related genes 30:56 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 25940448 7 34 from GAGs 1176:1179 arg1 Notch 1209:1213 Notch 1209:1213 Notch 1209:1213 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 35 theme biosynthetic 1151:1162 arg1 pathways 1164:1171 the biosynthetic pathways 1147:1171 the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans) 1147:1305 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 9 36 theme N-linked 1522:1529 arg1 glycans 1531:1537 terminal N-linked glycans 1513:1537 terminal N-linked glycans 1513:1537 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 2 37 theme glycosaminoglycans 329:346 arg1 region 319:324 the common core region 303:324 the common core region of glycosaminoglycans (GAGs) 303:353 For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. 25940448 4 38 theme genes 725:729 arg1 silencing 712:720 comprehensive silencing 698:720 comprehensive silencing of genes encoding GR and proteoglycan core proteins 698:772 To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. 25940448 5 39 theme N-linked 906:913 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 39 theme N-linked 906:913 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 2 40 theme common 307:312 arg1 region 319:324 the common core region 303:324 the common core region of glycosaminoglycans (GAGs) 303:353 For example, the common core region of glycosaminoglycans (GAGs) is sequentially synthesized by peptide-O-xylosyltransferase, β1,4-galactosyltransferase I, β1,3-galactosyltransferase II, and β1,3-glucuronyltransferase. 25940448 3 41 theme same 640:643 arg1 abnormality 656:666 the same phenotypic abnormality 636:666 the same phenotypic abnormality 636:666 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 9 42 theme terminal 1513:1520 arg1 glycans 1531:1537 terminal N-linked glycans 1513:1537 terminal N-linked glycans 1513:1537 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 5 43 theme O-linked 916:923 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 43 theme O-linked 916:923 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 44 theme functional 865:874 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 44 theme functional 865:874 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 45 theme GR 814:815 arg1 125 834:836 125 834:836 125 834:836 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 45 theme GR 814:815 arg1 genes 827:831 Drosophila GR candidate genes 803:831 Drosophila GR candidate genes (125) 803:837 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 1 46 theme glycan-modifying 229:244 arg1 enzymes 246:252 glycan-modifying enzymes 229:252 glycan-modifying enzymes 229:252 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 0 47 theme genes 52:56 arg1 clustering 16:25 Phenotype-based clustering 0:25 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 0:89 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 25940448 7 48 from glycan 1199:1204 arg1 Notch 1209:1213 Notch 1209:1213 Notch 1209:1213 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 5 49 theme candidate 817:825 arg1 125 834:836 125 834:836 125 834:836 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 49 theme candidate 817:825 arg1 genes 827:831 Drosophila GR candidate genes 803:831 Drosophila GR candidate genes (125) 803:837 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 9 50 theme galactosyltransferase 1466:1486 arg1 activity 1488:1495 galactosyltransferase activity 1466:1495 galactosyltransferase activity 1466:1495 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 3 51 theme functional 542:551 arg1 impairment 553:562 functional impairment 542:562 functional impairment of GR proteins involved in synthesis of the same glycan 542:618 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 0 52 theme gene 75:78 arg1 silencing 80:88 RNAi-mediated gene silencing 61:88 RNAi-mediated gene silencing 61:88 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 25940448 9 53 theme further 1419:1425 arg1 analysis 1427:1434 Our further analysis 1415:1434 Our further analysis 1415:1434 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 7 54 theme clustered 1116:1124 arg1 phenotypes 1126:1135 The clustered phenotypes 1112:1135 The clustered phenotypes 1112:1135 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 10 55 contain has 1574:1576 arg2 potential 1578:1586 potential 1578:1586 potential for the functional prediction of novel GR genes 1578:1634 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 10 55 contain has 1574:1576 arg1 clustering 1551:1560 Phenotypic clustering 1540:1560 Phenotypic clustering 1540:1560 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 0 56 theme RNAi-mediated 61:73 arg1 silencing 80:88 RNAi-mediated gene silencing 61:88 RNAi-mediated gene silencing 61:88 Phenotype-based clustering of glycosylation-related genes by RNAi-mediated gene silencing. 25940448 6 57 theme phenotypes 1027:1036 arg1 range 1008:1012 a range 1006:1012 a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation 1006:1109 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 1 58 theme glycosylation-related 163:183 arg1 enzymes 246:252 glycan-modifying enzymes 229:252 glycan-modifying enzymes 229:252 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 1 58 theme glycosylation-related 163:183 arg1 transporters 276:287 nucleotide-sugar transporters 259:287 nucleotide-sugar transporters 259:287 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 1 58 theme glycosylation-related 163:183 arg1 proteins 190:197 glycosylation-related (GR) proteins 163:197 glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters 163:287 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 1 58 theme glycosylation-related 163:183 arg1 glycosyltransferases 207:226 glycosyltransferases 207:226 glycosyltransferases 207:226 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 1 58 theme glycosylation-related 163:183 arg1 GR 186:187 GR 186:187 GR 186:187 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 6 59 theme malformed 1017:1025 arg1 phenotypes 1027:1036 malformed phenotypes 1017:1036 malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation 1017:1109 Spatiotemporally regulated silencing caused a range of malformed phenotypes that fell into three types: extra veins, thick veins, and depigmentation. 25940448 7 60 dep termed 1271:1276 arg1 herein 1264:1269 herein 1264:1269 herein 1264:1269 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 1 61 theme reactions 140:148 arg1 series 130:135 a series 128:135 a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters 128:287 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 10 62 theme functional 1596:1605 arg1 prediction 1607:1616 the functional prediction 1592:1616 the functional prediction of novel GR genes 1592:1634 Phenotypic clustering, therefore, has potential for the functional prediction of novel GR genes. 25940448 5 63 link N-linked 906:913 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 63 link N-linked 906:913 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 3 64 theme proteins 570:577 arg1 impairment 553:562 functional impairment 542:562 functional impairment of GR proteins involved in synthesis of the same glycan 542:618 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 8 65 theme terminal 1397:1404 arg1 domains 1406:1412 terminal domains 1397:1412 terminal domains 1397:1412 Based on the phenotypic clustering, CG33145 was predicted to be involved in formation of terminal domains. 25940448 3 66 theme GR 567:568 arg1 proteins 570:577 GR proteins 567:577 GR proteins involved in synthesis of the same glycan 567:618 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 7 67 theme glycans 1298:1304 arg1 glycans 1298:1304 glycans 1298:1304 glycans 1298:1304 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 7 67 theme glycans 1298:1304 arg1 domains 1287:1293 terminal domains 1278:1293 terminal domains of glycans 1278:1304 The clustered phenotypes reflected the biosynthetic pathways of GAGs, Fringe-dependent glycan on Notch, and glycans placed at or near nonreducing ends (herein termed terminal domains of glycans). 25940448 5 68 theme Notch-related 926:938 arg1 groups 876:881 five functional groups 860:881 five functional groups for synthesis of GAGs 860:903 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 5 68 theme Notch-related 926:938 arg1 glycans 953:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 N-linked, O-linked, Notch-related, and unknown glycans 906:959 Drosophila GR candidate genes (125) were classified into five functional groups for synthesis of GAGs, N-linked, O-linked, Notch-related, and unknown glycans. 25940448 9 69 theme glycans 1531:1537 arg1 synthesis 1500:1508 synthesis 1500:1508 synthesis of terminal N-linked glycans 1500:1537 Our further analysis showed that CG33145 exhibited galactosyltransferase activity in synthesis of terminal N-linked glycans. 25940448 1 70 theme nucleotide-sugar 259:274 arg1 transporters 276:287 nucleotide-sugar transporters 259:287 nucleotide-sugar transporters 259:287 Glycan structures are synthesized by a series of reactions conducted by glycosylation-related (GR) proteins such as glycosyltransferases, glycan-modifying enzymes, and nucleotide-sugar transporters. 25940448 3 71 theme same 608:611 arg1 glycan 613:618 the same glycan 604:618 the same glycan 604:618 This raises the possibility that functional impairment of GR proteins involved in synthesis of the same glycan might result in the same phenotypic abnormality. 25940448 4 72 theme comprehensive 698:710 arg1 silencing 712:720 comprehensive silencing 698:720 comprehensive silencing of genes encoding GR and proteoglycan core proteins 698:772 To examine this possibility, comprehensive silencing of genes encoding GR and proteoglycan core proteins was conducted in Drosophila. 27124023 0 0 theme Multiple 78:85 arg1 Study 110:114 A Multiple Ethnic Cross-Sectional Study 76:114 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 1 1 theme proteins 145:152 arg1 half 127:130 half 127:130 half of all known proteins 127:152 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 5 2 theme glycome 1276:1282 arg1 composition 1284:1294 IgG glycome composition 1272:1294 IgG glycome composition 1272:1294 The present study demonstrated for the 1st time an association between hypertension and IgG glycome composition. 27124023 2 3 theme possible 467:474 arg1 association 476:486 the possible association 463:486 the possible association between hypertension and IgG glycosylation 463:529 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 3 4 theme individuals 691:701 arg1 total 677:681 a total 675:681 a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 675:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 5 theme normal 1045:1050 arg1 pressure 1058:1065 normal blood pressure 1045:1065 normal blood pressure 1045:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 2 6 theme human 346:350 arg1 diseases.A 352:361 at least 30 severe human diseases.A 327:361 at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation 327:529 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 5 7 theme 1st 1223:1225 arg1 time 1227:1230 the 1st time 1219:1230 the 1st time 1219:1230 The present study demonstrated for the 1st time an association between hypertension and IgG glycome composition. 27124023 0 8 theme Cross-Sectional 94:108 arg1 Study 110:114 A Multiple Ethnic Cross-Sectional Study 76:114 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 3 9 theme blood 1052:1056 arg1 pressure 1058:1065 normal blood pressure 1045:1065 normal blood pressure 1045:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 1 10 dep half 127:130 arg1 More 117:120 More 117:120 More 117:120 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 3 11 theme normal 900:905 arg1 pressure 913:920 normal blood pressure 900:920 normal blood pressure 900:920 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 0 12 theme Ethnic 87:92 arg1 Study 110:114 A Multiple Ethnic Cross-Sectional Study 76:114 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 2 13 theme glycosylation 279:291 arg1 pathways 293:300 glycosylation pathways 279:300 glycosylation pathways 279:300 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 3 14 theme additional 929:938 arg1 traits 950:955 additional 17 glycan traits 929:955 additional 17 glycan traits 929:955 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 15 with participants 993:1004 arg1 hypertension 1011:1022 hypertension 1011:1022 hypertension 1011:1022 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 2 16 theme IgG 513:515 arg1 glycosylation 517:529 IgG glycosylation 513:529 IgG glycosylation 513:529 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 3 17 used used 610:613 arg2 chromatography 558:571 A hydrophilic interaction chromatography 532:571 A hydrophilic interaction chromatography of fluorescently labeled glycans 532:604 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 4 18 theme diastolic 1153:1161 arg1 DBP 1179:1181 DBP 1179:1181 DBP 1179:1181 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 4 18 theme diastolic 1153:1161 arg1 pressure 1169:1176 diastolic blood pressure 1153:1176 diastolic blood pressure (DBP) 1153:1182 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 2 19 dep diseases.A 352:361 arg1 carried 436:442 carried 436:442 was carried out to investigate the possible association between hypertension and IgG glycosylation 432:529 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 3 20 theme hydrophilic 534:544 arg1 chromatography 558:571 A hydrophilic interaction chromatography 532:571 A hydrophilic interaction chromatography of fluorescently labeled glycans 532:604 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 21 theme Croatian 719:726 arg1 individuals 691:701 4757 individuals 686:701 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 686:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 22 theme labeled 590:596 arg1 glycans 598:604 fluorescently labeled glycans 576:604 fluorescently labeled glycans 576:604 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 23 from IgG 648:650 arg1 samples 662:668 plasma samples 655:668 plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 655:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 23 from IgG 648:650 arg1 total 677:681 a total 675:681 a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 675:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 1 24 theme all 166:168 arg1 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins 117:205 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 3 25 dep glycans 757:763 arg1 IgG 766:768 IgG 766:768 IgG with digalactosylated glycans 766:798 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 5 26 theme present 1188:1194 arg1 study 1196:1200 The present study 1184:1200 The present study 1184:1200 The present study demonstrated for the 1st time an association between hypertension and IgG glycome composition. 27124023 3 27 theme glycans 598:604 arg1 chromatography 558:571 A hydrophilic interaction chromatography 532:571 A hydrophilic interaction chromatography of fluorescently labeled glycans 532:604 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 2 28 theme cross-sectional 379:393 arg1 study 395:399 multiple center cross-sectional study 363:399 multiple center cross-sectional study (China, Croatia, and Scotland) 363:430 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 4 29 theme blood 1163:1167 arg1 DBP 1179:1181 DBP 1179:1181 DBP 1179:1181 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 4 29 theme blood 1163:1167 arg1 pressure 1169:1176 diastolic blood pressure 1153:1176 diastolic blood pressure (DBP) 1153:1182 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 3 30 with IgG 766:768 arg1 glycans 792:798 digalactosylated glycans 775:798 digalactosylated glycans 775:798 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 6 31 theme anti-inflammatory 1462:1478 arg1 pathways 1480:1487 pro- and/or anti-inflammatory pathways 1450:1487 pro- and/or anti-inflammatory pathways 1450:1487 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 3 32 theme Scottish 733:740 arg1 individuals 691:701 4757 individuals 686:701 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 686:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 6 33 theme IgG 1372:1374 arg1 N-glycosylation 1353:1367 N-glycosylation 1353:1367 N-glycosylation of IgG 1353:1374 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 3 34 theme glycan 943:948 arg1 traits 950:955 additional 17 glycan traits 929:955 additional 17 glycan traits 929:955 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 1 35 theme extra-cellular 183:196 arg1 proteins 198:205 More than half of all known proteins, and almost all membrane and extra-cellular proteins 117:205 proteins 198:205 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 0 36 theme Immunoglobulin 41:54 arg1 G 56:56 Immunoglobulin G 41:56 Immunoglobulin G 41:56 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 3 37 theme ethnicity.Five 742:755 arg1 glycans 757:763 ethnicity.Five glycans 742:763 ethnicity.Five glycans (IgG with digalactosylated glycans) 742:799 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 38 from total 677:681 arg1 samples 662:668 plasma samples 655:668 plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 655:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 38 from total 677:681 arg1 IgG 648:650 IgG 648:650 IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 648:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 6 39 gly N-glycosylation 1353:1367 arg1 IgG 1372:1374 IgG 1372:1374 IgG 1372:1374 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 6 40 theme pro- 1450:1453 arg1 pathways 1480:1487 pro- and/or anti-inflammatory pathways 1450:1487 pro- and/or anti-inflammatory pathways 1450:1487 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 3 41 theme digalactosylated 775:790 arg1 glycans 792:798 digalactosylated glycans 775:798 digalactosylated glycans 775:798 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 42 with participants 827:838 arg1 hypertension 864:875 hypertension 864:875 hypertension 864:875 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 42 with participants 827:838 arg1 prehypertension 845:859 prehypertension 845:859 prehypertension 845:859 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 43 dep Croatian 719:726 arg1 differed 815:822 differed 815:822 differed 815:822 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 6 44 theme hypertension 1407:1418 arg1 pathogenesis 1391:1402 pathogenesis 1391:1402 pathogenesis of hypertension 1391:1418 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 3 45 theme plasma 655:660 arg1 samples 662:668 plasma samples 655:668 plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 655:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 2 46 theme severe 339:344 arg1 diseases.A 352:361 at least 30 severe human diseases.A 327:361 at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation 327:529 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 0 47 theme G 56:56 arg1 Association 4:14 The Association 0:14 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 1 48 contain have 207:210 arg1 proteins 198:205 More than half of all known proteins, and almost all membrane and extra-cellular proteins 117:205 proteins 198:205 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 1 48 contain have 207:210 arg1 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins 117:205 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 1 48 contain have 207:210 arg2 glycans 242:248 glycans 242:248 glycans attached to them 242:265 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 1 48 contain have 207:210 arg2 structures 228:237 oligosaccharide structures 212:237 oligosaccharide structures 212:237 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 4 49 theme systolic 1120:1127 arg1 SBP 1145:1147 SBP 1145:1147 SBP 1145:1147 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 4 49 theme systolic 1120:1127 arg1 pressure 1135:1142 systolic blood pressure 1120:1142 systolic blood pressure (SBP) 1120:1148 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 6 50 from variation 1340:1348 arg1 N-glycosylation 1353:1367 N-glycosylation 1353:1367 N-glycosylation of IgG 1353:1374 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 6 51 theme individual 1329:1338 arg1 variation 1340:1348 the individual variation 1325:1348 the individual variation in N-glycosylation of IgG 1325:1374 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 6 52 from effect 1440:1445 arg1 pathways 1480:1487 pro- and/or anti-inflammatory pathways 1450:1487 pro- and/or anti-inflammatory pathways 1450:1487 These findings suggest that the individual variation in N-glycosylation of IgG contributes to pathogenesis of hypertension, presumably via its effect on pro- and/or anti-inflammatory pathways. 27124023 2 53 dep study 395:399 arg1 Croatia 409:415 Croatia 409:415 Croatia 409:415 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 2 53 dep study 395:399 arg1 Scotland 422:429 Scotland 422:429 Scotland 422:429 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 2 53 dep study 395:399 arg1 China 402:406 China 402:406 China 402:406 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 4 54 dep significant 1092:1102 arg1 correlated 1104:1113 correlated 1104:1113 correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP) 1104:1182 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 1 55 theme half 127:130 arg1 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins 117:205 membrane 170:177 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 2 56 from Defects 268:274 arg1 pathways 293:300 glycosylation pathways 279:300 glycosylation pathways 279:300 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 1 57 theme oligosaccharide 212:226 arg1 structures 228:237 oligosaccharide structures 212:237 oligosaccharide structures 212:237 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 1 58 attach attached 250:257 arg1 them 262:265 them 262:265 them 262:265 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 1 58 attach attached 250:257 arg2 glycans 242:248 glycans 242:248 glycans attached to them 242:265 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 2 59 theme multiple 363:370 arg1 study 395:399 multiple center cross-sectional study 363:399 multiple center cross-sectional study (China, Croatia, and Scotland) 363:430 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 3 60 attach attached 636:643 arg2 N-glycans 626:634 N-glycans 626:634 N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 626:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 3 60 attach attached 636:643 arg1 IgG 648:650 IgG 648:650 IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure 648:1065 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 0 61 dep Association 4:14 arg1 Study 110:114 A Multiple Ethnic Cross-Sectional Study 76:114 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 5 62 theme IgG 1272:1274 arg1 composition 1284:1294 IgG glycome composition 1272:1294 IgG glycome composition 1272:1294 The present study demonstrated for the 1st time an association between hypertension and IgG glycome composition. 27124023 2 63 theme center 372:377 arg1 study 395:399 multiple center cross-sectional study 363:399 multiple center cross-sectional study (China, Croatia, and Scotland) 363:430 Defects in glycosylation pathways are directly involved in at least 30 severe human diseases.A multiple center cross-sectional study (China, Croatia, and Scotland) was carried out to investigate the possible association between hypertension and IgG glycosylation. 27124023 4 64 theme blood 1129:1133 arg1 SBP 1145:1147 SBP 1145:1147 SBP 1145:1147 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 4 64 theme blood 1129:1133 arg1 pressure 1135:1142 systolic blood pressure 1120:1142 systolic blood pressure (SBP) 1120:1148 These glycans were also significant correlated with systolic blood pressure (SBP) or diastolic blood pressure (DBP). 27124023 0 65 theme Hypertension 62:73 arg1 Association 4:14 The Association 0:14 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 0:115 The Association Between Glycosylation of Immunoglobulin G and Hypertension: A Multiple Ethnic Cross-Sectional Study. 27124023 3 66 theme interaction 546:556 arg1 chromatography 558:571 A hydrophilic interaction chromatography 532:571 A hydrophilic interaction chromatography of fluorescently labeled glycans 532:604 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 27124023 1 67 theme known 139:143 arg1 proteins 145:152 all known proteins 135:152 all known proteins 135:152 More than half of all known proteins, and almost all membrane and extra-cellular proteins have oligosaccharide structures or glycans attached to them. 27124023 3 68 theme blood 907:911 arg1 pressure 913:920 normal blood pressure 900:920 normal blood pressure 900:920 A hydrophilic interaction chromatography of fluorescently labeled glycans was used to analyze N-glycans attached to IgG in plasma samples from a total of 4757 individuals of Chinese Han, Croatian, and Scottish ethnicity.Five glycans (IgG with digalactosylated glycans) significantly differed in participants with prehypertension or hypertension compared to those with normal blood pressure, while additional 17 glycan traits were only significantly differed in participants with hypertension compared to those of normal blood pressure. 26646445 6 0 theme least 1104:1108 arg1 squares 1110:1116 partial least squares 1096:1116 partial least squares discriminant analysis 1096:1138 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 9 1 theme cancer 1649:1654 arg1 recurrence 1656:1665 breast cancer recurrence 1642:1665 breast cancer recurrence 1642:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 7 2 theme improved 1272:1279 arg1 performance 1292:1302 an improved diagnostic performance 1269:1302 an improved diagnostic performance 1269:1302 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 2 3 theme breast 272:277 arg1 patients 286:293 breast cancer patients 272:293 breast cancer patients 272:293 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 4 4 theme Lewis 632:636 arg1 X 638:638 Lewis X 632:638 Lewis X 632:638 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 6 5 theme partial 1096:1102 arg1 squares 1110:1116 partial least squares 1096:1116 partial least squares discriminant analysis 1096:1138 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 5 6 theme ion-trap 827:834 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 6 theme ion-trap 827:834 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 7 theme ionization 860:869 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 7 theme ionization 860:869 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 8 8 theme CA 1482:1483 arg1 15-3 1485:1488 CA 15-3 1482:1488 CA 15-3 1482:1488 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 3 9 with pattern 430:436 arg1 antennary 484:492 antennary 484:492 antennary 484:492 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 3 9 with pattern 430:436 arg1 core 475:478 core 475:478 core 475:478 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 5 10 theme sensitive 920:928 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 10 theme sensitive 920:928 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 2 11 gly bifucosylated 205:217 arg1 N-glycans 219:227 bifucosylated N-glycans 205:227 bifucosylated N-glycans 205:227 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 5 12 theme useful 934:939 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 12 theme useful 934:939 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 0 13 theme value 67:71 arg1 level 9:13 Elevated level 0:13 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 0:97 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 6 14 theme profile 1015:1021 arg1 data 1023:1026 MS profile data 1012:1026 MS profile data 1012:1026 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 8 15 theme bifucosylated 1366:1378 arg1 N-glycan 1380:1387 bifucosylated N-glycan 1366:1387 bifucosylated N-glycan 1366:1387 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 0 16 theme Chinese 76:82 arg1 cancer 91:96 Chinese breast cancer 76:96 Chinese breast cancer 76:96 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 7 17 theme CEA 1254:1256 arg1 levels 1258:1263 CEA levels 1254:1263 CEA levels 1254:1263 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 5 18 gly glycoproteins 721:733 arg1 glycoproteins 721:733 the glycoproteins 717:733 the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls 717:798 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 2 19 dep recurrence 366:375 arg1 the 362:364 the 362:364 the 362:364 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 2 20 theme breast 395:400 arg1 cancer 402:407 breast cancer 395:407 breast cancer 395:407 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 7 21 theme bifucosylation 1228:1241 arg1 degree 1243:1248 bifucosylation degree 1228:1248 bifucosylation degree 1228:1248 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 5 22 theme N-glycans 989:997 arg1 identification 971:984 identification 971:984 identification 971:984 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 22 theme N-glycans 989:997 arg1 detection 957:965 detection 957:965 detection 957:965 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 8 23 theme breast 1408:1413 arg1 recurrence 1422:1431 breast cancer recurrence 1408:1431 breast cancer recurrence 1408:1431 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 3 24 theme structural 419:428 arg1 pattern 430:436 A unique structural pattern 410:436 A unique structural pattern 410:436 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 0 25 theme Elevated 0:7 arg1 level 9:13 Elevated level 0:13 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 0:97 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 8 26 theme recurrence 1422:1431 arg1 potential 1353:1361 the potential 1349:1361 the potential of bifucosylated N-glycan 1349:1387 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 8 26 theme recurrence 1422:1431 arg1 indicator 1395:1403 an indicator 1392:1403 an indicator of breast cancer recurrence 1392:1431 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 3 27 theme breast 526:531 arg1 patients 540:547 breast cancer patients 526:547 breast cancer patients 526:547 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 3 28 theme N-glycan 455:462 arg1 pattern 430:436 A unique structural pattern 410:436 A unique structural pattern 410:436 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 0 29 theme glycoprotein 24:35 arg1 bifucosylation 37:50 serum glycoprotein bifucosylation 18:50 serum glycoprotein bifucosylation 18:50 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 9 30 theme N-glycans 1594:1602 arg1 indicator 1629:1637 a more reliable indicator 1613:1637 a more reliable indicator of breast cancer recurrence 1613:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 9 30 theme N-glycans 1594:1602 arg1 degree 1584:1589 the bifucosylation degree 1565:1589 the bifucosylation degree of N-glycans 1565:1602 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 6 31 theme statistical 1037:1047 arg1 tools 1049:1053 several statistical tools 1029:1053 several statistical tools 1029:1053 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 8 32 theme clinical 1450:1457 arg1 biomarkers 1459:1468 the current clinical biomarkers 1438:1468 the current clinical biomarkers 1438:1468 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 5 33 theme healthy 783:789 arg1 controls 791:798 43 healthy controls 780:798 43 healthy controls 780:798 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 6 34 theme receiver-operating 1144:1161 arg1 curve 1178:1182 receiver-operating characteristic curve 1144:1182 receiver-operating characteristic curve 1144:1182 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 3 35 gly bifucosylated 441:453 arg1 N-glycan 455:462 bifucosylated N-glycan 441:462 bifucosylated N-glycan 441:462 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 3 36 theme cancer 533:538 arg1 patients 540:547 breast cancer patients 526:547 breast cancer patients 526:547 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 6 37 theme squares 1110:1116 arg1 analysis 1131:1138 partial least squares discriminant analysis 1096:1138 partial least squares discriminant analysis 1096:1138 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 4 38 theme standard 612:619 arg1 spectra 621:627 the standard spectra 608:627 the standard spectra of Lewis X and H2 608:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 5 39 theme breast 753:758 arg1 patients 767:774 91 breast cancer patients 750:774 91 breast cancer patients 750:774 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 7 40 contain had 1265:1267 arg2 performance 1292:1302 an improved diagnostic performance 1269:1302 an improved diagnostic performance 1269:1302 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 7 40 contain had 1265:1267 arg1 measurement 1213:1223 the measurement 1209:1223 the measurement of bifucosylation degree and CEA levels 1209:1263 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 9 41 theme breast 1642:1647 arg1 recurrence 1656:1665 breast cancer recurrence 1642:1665 breast cancer recurrence 1642:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 7 42 theme diagnostic 1281:1290 arg1 performance 1292:1302 an improved diagnostic performance 1269:1302 an improved diagnostic performance 1269:1302 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 4 43 theme fucosylation 576:587 arg1 spectrum 554:561 The spectrum 550:561 The spectrum of antennary fucosylation 550:587 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 4 43 theme fucosylation 576:587 arg1 composite 595:603 a composite 593:603 a composite of the standard spectra of Lewis X and H2 593:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 4 43 theme fucosylation 576:587 arg1 spectra 621:627 the standard spectra 608:627 the standard spectra of Lewis X and H2 608:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 9 44 theme recurrence 1656:1665 arg1 degree 1584:1589 the bifucosylation degree 1565:1589 the bifucosylation degree of N-glycans 1565:1602 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 9 44 theme recurrence 1656:1665 arg1 indicator 1629:1637 a more reliable indicator 1613:1637 a more reliable indicator of breast cancer recurrence 1613:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 2 45 theme cancer 279:284 arg1 patients 286:293 breast cancer patients 272:293 breast cancer patients 272:293 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 5 46 theme mass 871:874 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 46 theme mass 871:874 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 4 47 theme H2 644:645 arg1 spectra 621:627 the standard spectra 608:627 the standard spectra of Lewis X and H2 608:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 5 48 theme cancer 760:765 arg1 patients 767:774 91 breast cancer patients 750:774 91 breast cancer patients 750:774 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 7 49 theme levels 1258:1263 arg1 measurement 1213:1223 the measurement 1209:1223 the measurement of bifucosylation degree and CEA levels 1209:1263 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 5 50 theme linear 820:825 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 50 theme linear 820:825 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 51 theme Permethylated 690:702 arg1 N-glycans 704:712 Permethylated N-glycans 690:712 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls 690:798 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 52 theme quadrupole-electrospray 836:858 arg1 spectrometry 876:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 linear ion-trap quadrupole-electrospray ionization mass spectrometry 820:887 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 52 theme quadrupole-electrospray 836:858 arg1 approach 941:948 a highly sensitive and useful approach 911:948 a highly sensitive and useful approach in the detection and identification of N-glycans 911:997 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 4 53 theme X 638:638 arg1 spectra 621:627 the standard spectra 608:627 the standard spectra of Lewis X and H2 608:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 4 54 theme epitopes 680:687 arg1 mixture 661:667 a mixture 659:667 a mixture of the two epitopes 659:687 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 6 55 theme several 1029:1035 arg1 tools 1049:1053 several statistical tools 1029:1053 several statistical tools 1029:1053 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 9 56 theme CA 1546:1547 arg1 15-3 1549:1552 CA 15-3 1546:1552 CA 15-3 1546:1552 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 2 57 theme healthy 251:257 arg1 controls 259:266 healthy controls 251:266 healthy controls 251:266 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 1 58 theme cancer 148:153 arg1 progression 155:165 cancer progression 148:165 cancer progression 148:165 Aberrant glycosylation is highly associated with cancer progression. 26646445 9 59 theme bifucosylation 1569:1582 arg1 indicator 1629:1637 a more reliable indicator 1613:1637 a more reliable indicator of breast cancer recurrence 1613:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 9 59 theme bifucosylation 1569:1582 arg1 degree 1584:1589 the bifucosylation degree 1565:1589 the bifucosylation degree of N-glycans 1565:1602 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 5 60 dep detection 957:965 arg1 the 953:955 the 953:955 the 953:955 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 0 61 theme breast 84:89 arg1 cancer 91:96 Chinese breast cancer 76:96 Chinese breast cancer 76:96 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 6 62 theme MS 1012:1013 arg1 data 1023:1026 MS profile data 1012:1026 MS profile data 1012:1026 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 0 63 from level 9:13 arg1 cancer 91:96 Chinese breast cancer 76:96 Chinese breast cancer 76:96 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 7 64 theme degree 1243:1248 arg1 measurement 1213:1223 the measurement 1209:1223 the measurement of bifucosylation degree and CEA levels 1209:1263 The results showed that the measurement of bifucosylation degree and CEA levels had an improved diagnostic performance compared with that of CEA alone. 26646445 3 65 dep core 475:478 arg1 fucosylation 494:505 fucosylation 494:505 fucosylation 494:505 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 8 66 theme N-glycan 1380:1387 arg1 potential 1353:1361 the potential 1349:1361 the potential of bifucosylated N-glycan 1349:1387 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 8 66 theme N-glycan 1380:1387 arg1 indicator 1395:1403 an indicator 1392:1403 an indicator of breast cancer recurrence 1392:1431 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 5 67 theme glycoproteins 721:733 arg1 N-glycans 704:712 Permethylated N-glycans 690:712 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls 690:798 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 3 68 theme bifucosylated 441:453 arg1 N-glycan 455:462 bifucosylated N-glycan 441:462 bifucosylated N-glycan 441:462 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 8 69 gly bifucosylated 1366:1378 arg1 N-glycan 1380:1387 bifucosylated N-glycan 1366:1387 bifucosylated N-glycan 1366:1387 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 0 70 theme serum 18:22 arg1 bifucosylation 37:50 serum glycoprotein bifucosylation 18:50 serum glycoprotein bifucosylation 18:50 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 1 71 theme Aberrant 99:106 arg1 glycosylation 108:120 Aberrant glycosylation 99:120 Aberrant glycosylation 99:120 Aberrant glycosylation is highly associated with cancer progression. 26646445 4 72 theme antennary 566:574 arg1 fucosylation 576:587 antennary fucosylation 566:587 antennary fucosylation 566:587 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 2 73 theme study 184:188 arg1 aim 172:174 The aim 168:174 The aim of this study 168:188 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 8 74 theme cancer 1415:1420 arg1 recurrence 1422:1431 breast cancer recurrence 1408:1431 breast cancer recurrence 1408:1431 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 0 75 theme bifucosylation 37:50 arg1 level 9:13 Elevated level 0:13 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 0:97 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 3 76 theme unique 412:417 arg1 pattern 430:436 A unique structural pattern 410:436 A unique structural pattern 410:436 A unique structural pattern of bifucosylated N-glycan, with both core and antennary fucosylation, was identified in breast cancer patients. 26646445 2 77 theme cancer 402:407 arg1 metastasis 381:390 metastasis 381:390 metastasis 381:390 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 2 77 theme cancer 402:407 arg1 recurrence 366:375 recurrence 366:375 recurrence 366:375 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 0 78 theme prognostic 56:65 arg1 value 67:71 prognostic value 56:71 prognostic value 56:71 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 6 79 theme characteristic 1163:1176 arg1 curve 1178:1182 receiver-operating characteristic curve 1144:1182 receiver-operating characteristic curve 1144:1182 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 2 80 theme bifucosylated 205:217 arg1 N-glycans 219:227 bifucosylated N-glycans 205:227 bifucosylated N-glycans 205:227 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 8 81 theme current 1442:1448 arg1 biomarkers 1459:1468 the current clinical biomarkers 1438:1468 the current clinical biomarkers 1438:1468 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 2 82 theme potential 327:335 arg1 indicator 337:345 a potential indicator 325:345 a potential indicator for monitoring the recurrence and metastasis of breast cancer 325:407 The aim of this study was to compare bifucosylated N-glycans in sera obtained from healthy controls and breast cancer patients, with the goal of identifying a potential indicator for monitoring the recurrence and metastasis of breast cancer. 26646445 8 83 dep CEA 1477:1479 arg1 i.e. 1471:1474 i.e. 1471:1474 i.e. 1471:1474 We compared the potential of bifucosylated N-glycan as an indicator of breast cancer recurrence with the current clinical biomarkers, i.e., CEA, CA 15-3 and CA125. 26646445 0 84 gly glycoprotein 24:35 arg1 glycoprotein 24:35 serum glycoprotein bifucosylation 18:50 serum glycoprotein bifucosylation 18:50 Elevated level of serum glycoprotein bifucosylation and prognostic value in Chinese breast cancer. 26646445 5 85 from approach 941:948 arg1 identification 971:984 identification 971:984 identification 971:984 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 5 85 from approach 941:948 arg1 detection 957:965 detection 957:965 detection 957:965 Permethylated N-glycans of the glycoproteins extracted from 91 breast cancer patients and 43 healthy controls were detected using linear ion-trap quadrupole-electrospray ionization mass spectrometry, which appeared to be a highly sensitive and useful approach in the detection and identification of N-glycans. 26646445 9 86 theme reliable 1620:1627 arg1 degree 1584:1589 the bifucosylation degree 1565:1589 the bifucosylation degree of N-glycans 1565:1602 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 9 86 theme reliable 1620:1627 arg1 indicator 1629:1637 a more reliable indicator 1613:1637 a more reliable indicator of breast cancer recurrence 1613:1665 The result revealed that, compared with CEA, CA 15-3 and CA125, the bifucosylation degree of N-glycans could be a more reliable indicator of breast cancer recurrence. 26646445 6 87 theme discriminant 1118:1129 arg1 analysis 1131:1138 partial least squares discriminant analysis 1096:1138 partial least squares discriminant analysis 1096:1138 To evaluate MS profile data, several statistical tools were applied, including Student'st-test, partial least squares discriminant analysis and receiver-operating characteristic curve. 26646445 4 88 theme spectra 621:627 arg1 spectrum 554:561 The spectrum 550:561 The spectrum of antennary fucosylation 550:587 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 4 88 theme spectra 621:627 arg1 composite 595:603 a composite 593:603 a composite of the standard spectra of Lewis X and H2 593:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 26646445 4 88 theme spectra 621:627 arg1 spectra 621:627 the standard spectra 608:627 the standard spectra of Lewis X and H2 608:645 The spectrum of antennary fucosylation was a composite of the standard spectra of Lewis X and H2, indicating a mixture of the two epitopes. 25142936 7 0 theme processing 1207:1216 arg1 pathway 1218:1224 the non-classical glycan processing pathway 1182:1224 the non-classical glycan processing pathway of brain eNPP 6 1182:1240 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 7 0 theme processing 1207:1216 arg1 due 1249:1251 due 1249:1251 due 1249:1251 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 4 1 theme glycans 726:732 arg1 structures 664:673 hybrid and high mannose type structures 635:673 structures 664:673 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 1 theme glycans 726:732 arg1 reminiscent 676:686 reminiscent 676:686 reminiscent of processed mannose-6-phosphorylated glycans 676:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 1 2 theme alkaline 291:298 arg1 phosphodiesterase/pyrophosphatase 198:230 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 182:232 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 182:240 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 2 theme alkaline 291:298 arg1 C 318:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C 245:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney 245:378 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 5 3 theme bovine 812:817 arg1 s-eNPP6 825:831 bovine brain s-eNPP6 812:831 bovine brain s-eNPP6 812:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 8 4 theme brain 1545:1549 arg1 system 1531:1536 the cell-cell communication system 1503:1536 the cell-cell communication system of the brain 1503:1549 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 1 5 theme Ecto-nucleotide 182:196 arg1 phosphodiesterase/pyrophosphatase 198:230 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 182:232 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 182:240 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 5 theme Ecto-nucleotide 182:196 arg1 eNPP6 235:239 eNPP6 235:239 eNPP6 235:239 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 5 theme Ecto-nucleotide 182:196 arg1 C 318:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C 245:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney 245:378 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 6 theme lysophospholipase 300:316 arg1 phosphodiesterase/pyrophosphatase 198:230 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 182:232 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 182:240 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 6 theme lysophospholipase 300:316 arg1 C 318:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C 245:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney 245:378 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 7 7 theme brain 1229:1233 arg1 eNPP 1235:1238 brain eNPP 6 1229:1240 brain eNPP 6 1229:1240 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 6 8 theme brain 940:944 arg1 s-eNPP6 946:952 brain s-eNPP6 940:952 brain s-eNPP6 940:952 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 9 used occupied 1058:1065 arg2 sites 1048:1052 the glycosylation sites 1030:1052 the glycosylation sites 1030:1052 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 9 used occupied 1058:1065 arg2 one 1023:1025 one 1023:1025 one 1023:1025 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 10 from H-sensitive 985:995 arg1 s-eNPP6 946:952 brain s-eNPP6 940:952 brain s-eNPP6 940:952 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 11 gly glycosylation 1034:1046 arg2 sites 1048:1052 the glycosylation sites 1030:1052 the glycosylation sites 1030:1052 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 5 12 from kidney 921:926 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 7 13 theme glycan 1200:1205 arg1 pathway 1218:1224 the non-classical glycan processing pathway 1182:1224 the non-classical glycan processing pathway of brain eNPP 6 1182:1240 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 7 13 theme glycan 1200:1205 arg1 due 1249:1251 due 1249:1251 due 1249:1251 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 7 14 from pathway 1347:1353 arg1 brain 1367:1371 brain 1367:1371 brain 1367:1371 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 4 15 theme brain 596:600 arg1 s-eNPP6 602:608 brain s-eNPP6 596:608 brain s-eNPP6 596:608 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 3 16 theme identical 488:496 arg1 55 kDa 510:515 55 kDa 510:515 55 kDa joined by a disulfide bridge 510:544 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 3 16 theme identical 488:496 arg1 monomers 498:505 two identical monomers 484:505 two identical monomers of 55 kDa joined by a disulfide bridge 484:544 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 7 17 theme Golgi 1323:1327 arg1 pathway 1347:1353 an alternative Golgi glycan-processing pathway 1308:1353 an alternative Golgi glycan-processing pathway of eNPP6 in brain 1308:1371 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 5 18 theme site-specific 777:789 arg1 structures 798:807 the site-specific glycan structures 773:807 the site-specific glycan structures of bovine brain s-eNPP6 773:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 7 19 theme non-classical 1186:1198 arg1 pathway 1218:1224 the non-classical glycan processing pathway 1182:1224 the non-classical glycan processing pathway of brain eNPP 6 1182:1240 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 7 19 theme non-classical 1186:1198 arg1 due 1249:1251 due 1249:1251 due 1249:1251 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 6 20 theme dimer 1159:1163 arg1 interface 1165:1173 the dimer interface 1155:1173 the dimer interface 1155:1173 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 4 21 from structures 664:673 arg1 s-eNPP6 602:608 brain s-eNPP6 596:608 brain s-eNPP6 596:608 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 22 theme type 659:662 arg1 structures 664:673 hybrid and high mannose type structures 635:673 structures 664:673 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 22 theme type 659:662 arg1 reminiscent 676:686 reminiscent 676:686 reminiscent of processed mannose-6-phosphorylated glycans 676:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 22 theme type 659:662 arg1 N-glycans 614:622 the N-glycans 610:622 the N-glycans 610:622 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 0 23 theme non-classical 4:16 arg1 specific 131:138 specific 131:138 specific 131:138 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 23 theme non-classical 4:16 arg1 pathway 38:44 The non-classical N-glycan processing pathway 0:44 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 0:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 8 24 theme brain-specific 1388:1401 arg1 important 1486:1494 important 1486:1494 important 1486:1494 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 8 24 theme brain-specific 1388:1401 arg1 expression 1403:1412 The resulting brain-specific expression 1374:1412 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans 1374:1461 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 5 25 theme glycan 791:796 arg1 structures 798:807 the site-specific glycan structures 773:807 the site-specific glycan structures of bovine brain s-eNPP6 773:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 6 26 theme likely 1098:1103 arg1 located 1120:1126 located 1120:1126 located 1120:1126 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 26 theme likely 1098:1103 arg1 glycan 1090:1095 an endo H-sensitive glycan 1070:1095 an endo H-sensitive glycan 1070:1095 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 27 theme H-sensitive 1078:1088 arg1 located 1120:1126 located 1120:1126 located 1120:1126 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 27 theme H-sensitive 1078:1088 arg1 glycan 1090:1095 an endo H-sensitive glycan 1070:1095 an endo H-sensitive glycan 1070:1095 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 28 located located 1120:1126 arg1 cleft 1139:1143 the cleft 1135:1143 the cleft formed by the dimer interface 1135:1173 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 28 located located 1120:1126 arg2 glycan 1090:1095 an endo H-sensitive glycan 1070:1095 an endo H-sensitive glycan 1070:1095 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 28 located located 1120:1126 arg2 located 1120:1126 located 1120:1126 located 1120:1126 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 29 theme endo 1073:1076 arg1 located 1120:1126 located 1120:1126 located 1120:1126 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 29 theme endo 1073:1076 arg1 glycan 1090:1095 an endo H-sensitive glycan 1070:1095 an endo H-sensitive glycan 1070:1095 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 0 30 theme processing 27:36 arg1 specific 131:138 specific 131:138 specific 131:138 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 30 theme processing 27:36 arg1 pathway 38:44 The non-classical N-glycan processing pathway 0:44 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 0:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 8 31 theme resulting 1378:1386 arg1 important 1486:1494 important 1486:1494 important 1486:1494 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 8 31 theme resulting 1378:1386 arg1 expression 1403:1412 The resulting brain-specific expression 1374:1412 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans 1374:1461 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 5 32 theme glycan 872:877 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 0 33 theme N-glycan 18:25 arg1 specific 131:138 specific 131:138 specific 131:138 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 33 theme N-glycan 18:25 arg1 pathway 38:44 The non-classical N-glycan processing pathway 0:44 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 0:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 2 34 theme eNPP6 415:419 arg1 shedding 388:395 shedding 388:395 shedding of the GPI-anchor eNPP6 388:419 Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). 25142936 8 35 theme cell-cell 1507:1515 arg1 system 1531:1536 the cell-cell communication system 1503:1536 the cell-cell communication system of the brain 1503:1549 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 4 36 theme mannose-6-phosphorylated 701:724 arg1 glycans 726:732 processed mannose-6-phosphorylated glycans 691:732 processed mannose-6-phosphorylated glycans 691:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 37 theme hybrid 635:640 arg1 structures 664:673 hybrid and high mannose type structures 635:673 structures 664:673 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 37 theme hybrid 635:640 arg1 reminiscent 676:686 reminiscent 676:686 reminiscent of processed mannose-6-phosphorylated glycans 676:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 37 theme hybrid 635:640 arg1 N-glycans 614:622 the N-glycans 610:622 the N-glycans 610:622 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 5 38 theme brain 819:823 arg1 s-eNPP6 825:831 bovine brain s-eNPP6 812:831 bovine brain s-eNPP6 812:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 6 39 from s-eNPP6 946:952 arg1 H-sensitive 985:995 H-sensitive 985:995 H-sensitive 985:995 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 7 40 theme alternative 1311:1321 arg1 pathway 1347:1353 an alternative Golgi glycan-processing pathway 1308:1353 an alternative Golgi glycan-processing pathway of eNPP6 in brain 1308:1371 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 4 41 theme mannose 651:657 arg1 structures 664:673 hybrid and high mannose type structures 635:673 structures 664:673 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 41 theme mannose 651:657 arg1 reminiscent 676:686 reminiscent 676:686 reminiscent of processed mannose-6-phosphorylated glycans 676:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 41 theme mannose 651:657 arg1 N-glycans 614:622 the N-glycans 610:622 the N-glycans 610:622 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 42 theme processed 691:699 arg1 glycans 726:732 processed mannose-6-phosphorylated glycans 691:732 processed mannose-6-phosphorylated glycans 691:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 5 43 theme s-eNPP6 825:831 arg1 structures 798:807 the site-specific glycan structures 773:807 the site-specific glycan structures of bovine brain s-eNPP6 773:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 5 44 theme endo 853:856 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 5 45 theme s-eNPP6 891:897 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 4 46 theme high 646:649 arg1 structures 664:673 hybrid and high mannose type structures 635:673 structures 664:673 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 46 theme high 646:649 arg1 reminiscent 676:686 reminiscent 676:686 reminiscent of processed mannose-6-phosphorylated glycans 676:732 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 4 46 theme high 646:649 arg1 N-glycans 614:622 the N-glycans 610:622 the N-glycans 610:622 In brain s-eNPP6 the N-glycans are mainly hybrid and high mannose type structures, reminiscent of processed mannose-6-phosphorylated glycans. 25142936 0 47 theme brain 56:60 arg1 phosphodiesterase/pyrophosphatase 78:110 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 49:112 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 49:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 47 theme brain 56:60 arg1 eNPP6 115:119 eNPP6 115:119 eNPP6 115:119 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 1 48 theme myelin 362:367 arg1 brain 356:360 brain myelin 356:367 brain myelin 356:367 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 5 49 theme structures 798:807 arg1 characterization 753:768 characterization 753:768 characterization of the site-specific glycan structures of bovine brain s-eNPP6 753:831 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 8 50 theme accessible 1417:1426 arg1 hybrid 1428:1433 accessible hybrid 1417:1433 accessible hybrid 1417:1433 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 3 51 theme 55 kDa 510:515 arg1 55 kDa 510:515 55 kDa 510:515 55 kDa joined by a disulfide bridge 510:544 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 3 51 theme 55 kDa 510:515 arg1 monomers 498:505 two identical monomers 484:505 two identical monomers of 55 kDa joined by a disulfide bridge 484:544 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 0 52 theme bovine 49:54 arg1 phosphodiesterase/pyrophosphatase 78:110 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 49:112 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 49:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 52 theme bovine 49:54 arg1 eNPP6 115:119 eNPP6 115:119 eNPP6 115:119 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 8 53 theme oligomannosidic 1439:1453 arg1 glycans 1455:1461 oligomannosidic glycans 1439:1461 oligomannosidic glycans 1439:1461 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 7 54 theme glycan-processing 1329:1345 arg1 pathway 1347:1353 an alternative Golgi glycan-processing pathway 1308:1353 an alternative Golgi glycan-processing pathway of eNPP6 in brain 1308:1371 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 3 55 contain possesses 551:559 arg1 eNPP 465:468 eNPP 6 465:470 eNPP 6 465:470 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 3 55 contain possesses 551:559 arg2 N-glycans 566:574 four N-glycans 561:574 four N-glycans 561:574 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 7 56 theme eNPP6 1358:1362 arg1 pathway 1347:1353 an alternative Golgi glycan-processing pathway 1308:1353 an alternative Golgi glycan-processing pathway of eNPP6 in brain 1308:1371 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 6 57 dep likely 1098:1103 arg1 N406 1105:1108 N406 1105:1108 N406 1105:1108 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 58 theme sites 1048:1052 arg1 one 1023:1025 one 1023:1025 one 1023:1025 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 6 58 theme sites 1048:1052 arg1 sites 1048:1052 the glycosylation sites 1030:1052 the glycosylation sites 1030:1052 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 8 59 theme communication 1517:1529 arg1 system 1531:1536 the cell-cell communication system 1503:1536 the cell-cell communication system of the brain 1503:1549 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 8 60 theme hybrid 1428:1433 arg1 important 1486:1494 important 1486:1494 important 1486:1494 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 8 60 theme hybrid 1428:1433 arg1 expression 1403:1412 The resulting brain-specific expression 1374:1412 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans 1374:1461 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 5 61 from liver 911:915 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 6 62 theme glycosylation 1034:1046 arg1 sites 1048:1052 the glycosylation sites 1030:1052 the glycosylation sites 1030:1052 Whereas in brain s-eNPP6 all of the N-glycans were endo H-sensitive, in liver and kidney only one of the glycosylation sites was occupied by an endo H-sensitive glycan, likely N406, which is located within the cleft formed by the dimer interface. 25142936 0 63 theme phosphodiesterase/pyrophosphatase 78:110 arg1 specific 131:138 specific 131:138 specific 131:138 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 63 theme phosphodiesterase/pyrophosphatase 78:110 arg1 pathway 38:44 The non-classical N-glycan processing pathway 0:44 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 0:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 2 64 theme soluble 438:444 arg1 isoform 446:452 a soluble isoform 436:452 a soluble isoform (s-eNPP6) 436:462 Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). 25142936 2 64 theme soluble 438:444 arg1 s-eNPP6 455:461 s-eNPP6 455:461 s-eNPP6 455:461 Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). 25142936 8 65 theme glycans 1455:1461 arg1 important 1486:1494 important 1486:1494 important 1486:1494 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 8 65 theme glycans 1455:1461 arg1 expression 1403:1412 The resulting brain-specific expression 1374:1412 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans 1374:1461 The resulting brain-specific expression of accessible hybrid and oligomannosidic glycans may be physiologically important within the cell-cell communication system of the brain. 25142936 3 66 theme disulfide 529:537 arg1 bridge 539:544 a disulfide bridge 527:544 a disulfide bridge 527:544 eNPP 6 consists of two identical monomers of 55 kDa joined by a disulfide bridge, and possesses four N-glycans in each monomer. 25142936 5 67 theme bovine 904:909 arg1 liver 911:915 bovine liver 904:915 bovine liver 904:915 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 0 68 theme ecto-nucleotide 62:76 arg1 phosphodiesterase/pyrophosphatase 78:110 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 49:112 bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 49:120 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 0 68 theme ecto-nucleotide 62:76 arg1 eNPP6 115:119 eNPP6 115:119 eNPP6 115:119 The non-classical N-glycan processing pathway of bovine brain ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is brain specific and not due to mannose-6-phosphorylation. 25142936 5 69 theme H-sensitivity 858:870 arg1 profiles 879:886 the endo H-sensitivity glycan profiles 849:886 the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney 849:926 Here we completed characterization of the site-specific glycan structures of bovine brain s-eNPP6, and determined the endo H-sensitivity glycan profiles of s-eNPP6 from bovine liver and kidney. 25142936 2 70 theme GPI-anchor 404:413 arg1 eNPP6 415:419 the GPI-anchor eNPP6 400:419 the GPI-anchor eNPP6 400:419 Due to shedding of the GPI-anchor eNPP6 occurs also as a soluble isoform (s-eNPP6). 25142936 7 71 theme eNPP 1235:1238 arg1 pathway 1218:1224 the non-classical glycan processing pathway 1182:1224 the non-classical glycan processing pathway of brain eNPP 6 1182:1240 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 7 71 theme eNPP 1235:1238 arg1 due 1249:1251 due 1249:1251 due 1249:1251 Thus, the non-classical glycan processing pathway of brain eNPP 6 is not due to mannose-6-phosphorylation, suggesting that there is an alternative Golgi glycan-processing pathway of eNPP6 in brain. 25142936 1 72 theme -anchored 281:289 arg1 phosphodiesterase/pyrophosphatase 198:230 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 182:232 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) 182:240 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 25142936 1 72 theme -anchored 281:289 arg1 C 318:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C 245:318 a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney 245:378 Ecto-nucleotide phosphodiesterase/pyrophosphatase 6 (eNPP6) is a glycosylphosphatidylinositol (GPI)-anchored alkaline lysophospholipase C which is predominantly expressed in brain myelin and kidney. 24508870 10 0 link N-linked 1818:1825 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 1 1 theme chain 303:307 arg1 structures 309:318 unusual glycosylphosphatidylinositol-anchor side chain structures 254:318 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 10 2 theme complex 1769:1775 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 10 3 link GPI-linked 1831:1840 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 3 4 theme parasite 625:632 arg1 form 613:616 the mammalian bloodstream form 587:616 the mammalian bloodstream form of the parasite 587:632 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 5 5 theme molecular 1034:1042 arg1 weight 1044:1049 an approximate molecular weight 1019:1049 an approximate molecular weight of 110 kDa in the mutant lysate 1019:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 9 6 theme Immunofluorescence 1568:1585 arg1 microscopy 1587:1596 Immunofluorescence microscopy 1568:1596 Immunofluorescence microscopy 1568:1596 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 1 7 contain contain 325:331 arg2 units 380:384 branched N-acetyllactosamine and lacto-N-biose units 333:384 units 380:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 1 7 contain contain 325:331 arg1 structures 309:318 unusual glycosylphosphatidylinositol-anchor side chain structures 254:318 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 1 7 contain contain 325:331 arg2 N-acetyllactosamine 342:360 branched N-acetyllactosamine and lacto-N-biose units 333:384 N-acetyllactosamine 342:360 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 3 8 dep T. 681:682 arg1 brucei 684:689 brucei 684:689 brucei 684:689 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 4 9 theme reduced 774:780 arg1 binding 782:788 significantly reduced binding 760:788 significantly reduced binding 760:788 Under non-permissive conditions, the mutant showed significantly reduced binding to tomato lectin, which recognizes poly-N-acetyllactosamine-containing glycans. 24508870 3 10 theme conditional 664:674 arg1 mutant 691:696 a tetracycline-inducible conditional null T. brucei mutant 639:696 a tetracycline-inducible conditional null T. brucei mutant for TbGT8 639:706 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 9 11 theme epitope-tagged 1643:1656 arg1 TbGT8 1658:1662 hemagglutinin epitope-tagged TbGT8 1629:1662 hemagglutinin epitope-tagged TbGT8 1629:1662 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 6 12 theme Proteomic 1084:1092 arg1 analysis 1094:1101 Proteomic analysis 1084:1101 Proteomic analysis 1084:1101 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 10 13 theme type 1813:1816 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 0 14 theme GPI-anchor 115:124 arg1 glycan 136:141 a GPI-anchor modifying glycan 113:141 a GPI-anchor modifying glycan 113:141 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 1 15 theme surface 227:233 arg1 glycoproteins 235:247 procyclin surface glycoproteins 217:247 procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 217:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 0 16 from glycans 69:75 arg1 AcP115 102:107 a protein phosphatase AcP115 80:107 a protein phosphatase AcP115 80:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 8 17 theme protein 1457:1463 arg1 cores 1465:1469 the underlying protein cores 1442:1469 the underlying protein cores 1442:1469 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 8 17 theme protein 1457:1463 arg1 same 1480:1483 same 1480:1483 same 1480:1483 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 10 18 theme glycans 1842:1848 arg1 construction 1753:1764 the construction 1749:1764 the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1749:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 0 19 theme Trypanosoma 146:156 arg1 brucei 158:163 Trypanosoma brucei 146:163 Trypanosoma brucei 146:163 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 3 20 theme T. 681:682 arg1 mutant 691:696 a tetracycline-inducible conditional null T. brucei mutant 639:696 a tetracycline-inducible conditional null T. brucei mutant for TbGT8 639:706 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 7 21 from 10kDa 1366:1370 arg1 mutant 1387:1392 the mutant 1383:1392 the mutant 1383:1392 Western blotting with an anti-AcP115 antibody revealed that AcP115 was approximately 10kDa smaller in the mutant. 24508870 4 22 theme poly-N-acetyllactosamine-containing 825:859 arg1 glycans 861:867 poly-N-acetyllactosamine-containing glycans 825:867 poly-N-acetyllactosamine-containing glycans 825:867 Under non-permissive conditions, the mutant showed significantly reduced binding to tomato lectin, which recognizes poly-N-acetyllactosamine-containing glycans. 24508870 5 23 theme approximate 1022:1032 arg1 weight 1044:1049 an approximate molecular weight 1019:1049 an approximate molecular weight of 110 kDa in the mutant lysate 1019:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 5 24 with band 1009:1012 arg1 weight 1044:1049 an approximate molecular weight 1019:1049 an approximate molecular weight of 110 kDa in the mutant lysate 1019:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 6 25 contain contained 1126:1134 arg2 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 25 contain contained 1126:1134 arg1 band 1121:1124 the band 1117:1124 the band 1117:1124 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 25 contain contained 1126:1134 arg2 glycoproteins 1144:1156 several glycoproteins 1136:1156 several glycoproteins 1136:1156 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 0 26 theme phosphatase 90:100 arg1 AcP115 102:107 a protein phosphatase AcP115 80:107 a protein phosphatase AcP115 80:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 2 27 theme chain 467:471 arg1 synthesis 436:444 the synthesis 432:444 the synthesis of the branched side chain through its UDP-GlcNAc 432:494 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 5 28 theme pull-down 877:885 arg1 assays 887:892 Lectin pull-down assays 870:892 Lectin pull-down assays 870:892 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 2 29 theme branched 453:460 arg1 chain 467:471 the branched side chain 449:471 the branched side chain 449:471 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 5 30 theme null-mutant 947:957 arg1 TbGT8 941:945 TbGT8 941:945 TbGT8 null-mutant T. brucei 941:967 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 6 31 theme phosphatase 1193:1203 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 31 theme phosphatase 1193:1203 arg1 glycoprotein 1230:1241 a stage-specific glycoprotein 1213:1241 a stage-specific glycoprotein in the bloodstream form of T. brucei 1213:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 2 32 theme β1-3N-acetylglucosaminyltransferase 502:536 arg1 activity 538:545 βGal β1-3N-acetylglucosaminyltransferase activity 497:545 βGal β1-3N-acetylglucosaminyltransferase activity 497:545 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 5 33 theme mutant 1069:1074 arg1 lysate 1076:1081 the mutant lysate 1065:1081 the mutant lysate 1065:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 6 34 theme acidic 1173:1178 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 34 theme acidic 1173:1178 arg1 glycoprotein 1230:1241 a stage-specific glycoprotein 1213:1241 a stage-specific glycoprotein in the bloodstream form of T. brucei 1213:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 8 35 theme N-linked 1550:1557 arg1 glycans 1559:1565 N-linked glycans 1550:1565 N-linked glycans 1550:1565 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 10 36 theme parasite 1896:1903 arg1 forms 1905:1909 the bloodstream and procyclic parasite forms 1866:1909 forms 1905:1909 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 5 37 theme wild 927:930 arg1 type 932:935 the wild type 923:935 the wild type 923:935 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 1 38 theme lacto-N-biose 366:378 arg1 units 380:384 branched N-acetyllactosamine and lacto-N-biose units 333:384 units 380:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 8 39 link N-linked 1550:1557 arg1 glycans 1559:1565 N-linked glycans 1550:1565 N-linked glycans 1550:1565 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 8 40 theme Enzymatic 1395:1403 arg1 de-N-glycosylation 1405:1422 Enzymatic de-N-glycosylation 1395:1422 Enzymatic de-N-glycosylation 1395:1422 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 6 41 theme bloodstream 1250:1260 arg1 form 1262:1265 the bloodstream form 1246:1265 the bloodstream form of T. brucei 1246:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 0 42 from glycan 136:141 arg1 AcP115 102:107 a protein phosphatase AcP115 80:107 a protein phosphatase AcP115 80:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 3 43 theme mammalian 591:599 arg1 form 613:616 the mammalian bloodstream form 587:616 the mammalian bloodstream form of the parasite 587:632 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 1 44 theme unusual 254:260 arg1 structures 309:318 unusual glycosylphosphatidylinositol-anchor side chain structures 254:318 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 1 45 with glycoproteins 235:247 arg1 structures 309:318 unusual glycosylphosphatidylinositol-anchor side chain structures 254:318 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 1 46 theme side 298:301 arg1 chain 303:307 glycosylphosphatidylinositol-anchor side chain 262:307 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 6 47 theme stage-specific 1215:1228 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 47 theme stage-specific 1215:1228 arg1 glycoprotein 1230:1241 a stage-specific glycoprotein 1213:1241 a stage-specific glycoprotein in the bloodstream form of T. brucei 1213:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 10 48 theme poly-N-acetyllactosamine-containing 1777:1811 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 5 49 theme broad 995:999 arg1 band 1009:1012 a broad protein band 993:1012 a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate 993:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 1 50 theme procyclic 170:178 arg1 form 180:183 The procyclic form 166:183 The procyclic form of Trypanosoma brucei 166:205 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 0 51 link N-linked 60:67 arg1 glycans 69:75 N-linked glycans 60:75 N-linked glycans on a protein phosphatase AcP115 60:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 7 52 with blotting 1289:1296 arg1 antibody 1318:1325 an anti-AcP115 antibody 1303:1325 an anti-AcP115 antibody 1303:1325 Western blotting with an anti-AcP115 antibody revealed that AcP115 was approximately 10kDa smaller in the mutant. 24508870 10 53 theme GPI-linked 1831:1840 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 3 54 from role 570:573 arg1 form 613:616 the mammalian bloodstream form 587:616 the mammalian bloodstream form of the parasite 587:632 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 3 55 theme tetracycline-inducible 641:662 arg1 mutant 691:696 a tetracycline-inducible conditional null T. brucei mutant 639:696 a tetracycline-inducible conditional null T. brucei mutant for TbGT8 639:706 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 9 56 theme hemagglutinin 1629:1641 arg1 TbGT8 1658:1662 hemagglutinin epitope-tagged TbGT8 1629:1662 hemagglutinin epitope-tagged TbGT8 1629:1662 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 10 57 theme N-linked 1818:1825 arg1 glycans 1842:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans 1769:1848 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 0 58 theme modifying 126:134 arg1 glycan 136:141 a GPI-anchor modifying glycan 113:141 a GPI-anchor modifying glycan 113:141 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 9 59 theme TbGT8 1658:1662 arg1 colocalization 1611:1624 the colocalization 1607:1624 the colocalization of hemagglutinin epitope-tagged TbGT8 1607:1662 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 9 59 theme TbGT8 1658:1662 arg1 GRASP 1697:1701 the Golgi-associated protein GRASP 1668:1701 the Golgi-associated protein GRASP 1668:1701 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 6 60 theme brucei 1273:1278 arg1 form 1262:1265 the bloodstream form 1246:1265 the bloodstream form of T. brucei 1246:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 4 61 theme non-permissive 715:728 arg1 conditions 730:739 non-permissive conditions 715:739 non-permissive conditions 715:739 Under non-permissive conditions, the mutant showed significantly reduced binding to tomato lectin, which recognizes poly-N-acetyllactosamine-containing glycans. 24508870 1 62 theme Trypanosoma 188:198 arg1 brucei 200:205 Trypanosoma brucei 188:205 Trypanosoma brucei 188:205 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 1 63 theme procyclin 217:225 arg1 glycoproteins 235:247 procyclin surface glycoproteins 217:247 procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 217:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 3 64 theme null 676:679 arg1 mutant 691:696 a tetracycline-inducible conditional null T. brucei mutant 639:696 a tetracycline-inducible conditional null T. brucei mutant for TbGT8 639:706 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 9 65 theme protein 1689:1695 arg1 GRASP 1697:1701 the Golgi-associated protein GRASP 1668:1701 the Golgi-associated protein GRASP 1668:1701 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 8 66 theme underlying 1446:1455 arg1 cores 1465:1469 the underlying protein cores 1442:1469 the underlying protein cores 1442:1469 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 8 66 theme underlying 1446:1455 arg1 same 1480:1483 same 1480:1483 same 1480:1483 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 5 67 theme protein 1001:1007 arg1 band 1009:1012 a broad protein band 993:1012 a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate 993:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 1 68 theme brucei 200:205 arg1 form 180:183 The procyclic form 166:183 The procyclic form of Trypanosoma brucei 166:205 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 5 69 theme band 1009:1012 arg1 absence 982:988 the absence 978:988 the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate 978:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 5 69 theme band 1009:1012 arg1 differences 903:913 differences 903:913 differences between the wild type 903:935 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 5 69 theme band 1009:1012 arg1 TbGT8 941:945 TbGT8 941:945 TbGT8 null-mutant T. brucei 941:967 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 6 70 theme several 1136:1142 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 70 theme several 1136:1142 arg1 glycoproteins 1144:1156 several glycoproteins 1136:1156 several glycoproteins 1136:1156 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 0 71 theme protein 82:88 arg1 AcP115 102:107 a protein phosphatase AcP115 80:107 a protein phosphatase AcP115 80:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 4 72 theme tomato 793:798 arg1 lectin 800:805 tomato lectin 793:805 tomato lectin 793:805 Under non-permissive conditions, the mutant showed significantly reduced binding to tomato lectin, which recognizes poly-N-acetyllactosamine-containing glycans. 24508870 1 73 gly glycoproteins 235:247 arg1 glycoproteins 235:247 procyclin surface glycoproteins 217:247 procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 217:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 6 74 from glycoprotein 1230:1241 arg1 form 1262:1265 the bloodstream form 1246:1265 the bloodstream form of T. brucei 1246:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 75 gly glycoprotein 1230:1241 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 75 gly glycoprotein 1230:1241 arg1 glycoprotein 1230:1241 a stage-specific glycoprotein 1213:1241 a stage-specific glycoprotein in the bloodstream form of T. brucei 1213:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 5 76 theme Lectin 870:875 arg1 assays 887:892 Lectin pull-down assays 870:892 Lectin pull-down assays 870:892 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 8 77 theme 10-kDa 1506:1511 arg1 due 1528:1530 due 1528:1530 due 1528:1530 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 8 77 theme 10-kDa 1506:1511 arg1 difference 1513:1522 the 10-kDa difference 1502:1522 the 10-kDa difference 1502:1522 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 2 78 theme side 462:465 arg1 chain 467:471 the branched side chain 449:471 the branched side chain 449:471 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 5 79 theme kDa 1058:1060 arg1 weight 1044:1049 an approximate molecular weight 1019:1049 an approximate molecular weight of 110 kDa in the mutant lysate 1019:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 0 80 theme bifunctional 11:22 arg1 TbGT8 0:4 TbGT8 0:4 TbGT8 0:4 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 0 80 theme bifunctional 11:22 arg1 glycosyltransferase 24:42 a bifunctional glycosyltransferase 9:42 a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei 9:163 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 1 81 theme branched 333:340 arg1 N-acetyllactosamine 342:360 branched N-acetyllactosamine and lacto-N-biose units 333:384 N-acetyllactosamine 342:360 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 24508870 5 82 from weight 1044:1049 arg1 lysate 1076:1081 the mutant lysate 1065:1081 the mutant lysate 1065:1081 Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110 kDa in the mutant lysate. 24508870 6 83 theme ecto-protein 1180:1191 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 83 theme ecto-protein 1180:1191 arg1 glycoprotein 1230:1241 a stage-specific glycoprotein 1213:1241 a stage-specific glycoprotein in the bloodstream form of T. brucei 1213:1278 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 2 84 theme βGal 497:500 arg1 activity 538:545 βGal β1-3N-acetylglucosaminyltransferase activity 497:545 βGal β1-3N-acetylglucosaminyltransferase activity 497:545 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 7 85 dep 10kDa 1366:1370 arg1 smaller 1372:1378 smaller 1372:1378 smaller 1372:1378 Western blotting with an anti-AcP115 antibody revealed that AcP115 was approximately 10kDa smaller in the mutant. 24508870 10 86 theme forms 1905:1909 arg1 Golgi 1857:1861 the Golgi 1853:1861 the Golgi of the bloodstream and procyclic parasite forms, respectively 1853:1923 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 7 87 theme Western 1281:1287 arg1 blotting 1289:1296 Western blotting 1281:1296 Western blotting with an anti-AcP115 antibody 1281:1325 Western blotting with an anti-AcP115 antibody revealed that AcP115 was approximately 10kDa smaller in the mutant. 24508870 10 88 theme procyclic 1886:1894 arg1 forms 1905:1909 the bloodstream and procyclic parasite forms 1866:1909 forms 1905:1909 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 0 89 theme N-linked 60:67 arg1 glycans 69:75 N-linked glycans 60:75 N-linked glycans on a protein phosphatase AcP115 60:107 TbGT8 is a bifunctional glycosyltransferase that elaborates N-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in Trypanosoma brucei. 24508870 3 90 theme TbGT8 578:582 arg1 role 570:573 the role 566:573 the role of TbGT8 in the mammalian bloodstream form of the parasite 566:632 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 6 91 gly glycoproteins 1144:1156 arg1 AcP115 1205:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 the acidic ecto-protein phosphatase AcP115 1169:1210 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 6 91 gly glycoproteins 1144:1156 arg1 glycoproteins 1144:1156 several glycoproteins 1136:1156 several glycoproteins 1136:1156 Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. 24508870 2 92 theme glycosyltransferase 391:409 arg1 TbGT8 411:415 The glycosyltransferase TbGT8 387:415 The glycosyltransferase TbGT8 387:415 The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. 24508870 9 93 theme Golgi-associated 1672:1687 arg1 GRASP 1697:1701 the Golgi-associated protein GRASP 1668:1701 the Golgi-associated protein GRASP 1668:1701 Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. 24508870 10 94 theme bloodstream 1870:1880 arg1 Golgi 1857:1861 the Golgi 1853:1861 the Golgi of the bloodstream and procyclic parasite forms, respectively 1853:1923 These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively. 24508870 3 95 theme bloodstream 601:611 arg1 form 613:616 the mammalian bloodstream form 587:616 the mammalian bloodstream form of the parasite 587:632 Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. 24508870 8 96 from differences 1535:1545 arg1 glycans 1559:1565 N-linked glycans 1550:1565 N-linked glycans 1550:1565 Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. 24508870 1 97 theme glycosylphosphatidylinositol-anchor 262:296 arg1 chain 303:307 glycosylphosphatidylinositol-anchor side chain 262:307 unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units 254:384 The procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. 28905229 7 0 theme increased 1436:1444 arg1 risk 1446:1449 increased risk 1436:1449 increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations) 1436:1547 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 1 theme protein 1777:1783 arg1 N-glycans 1785:1793 plasma protein N-glycans 1770:1793 plasma protein N-glycans 1770:1793 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 4 2 theme greater 578:584 arg1 risk 586:589 greater risk 578:589 greater risk of developing type 2 diabetes 578:619 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 2 3 from Changes 184:190 arg1 N-glycome 208:216 human plasma N-glycome 195:216 human plasma N-glycome 195:216 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 9 4 theme further 2035:2041 arg1 research 2043:2050 further research 2035:2050 further research 2035:2050 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 2 5 theme diagnostic 276:285 arg1 biomarkers 302:311 promising diagnostic and prognostic biomarkers 266:311 promising diagnostic and prognostic biomarkers 266:311 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 5 6 from proteins 705:712 arg1 glycans 685:691 N-linked glycans 676:691 N-linked glycans from plasma proteins 676:712 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 8 7 theme Increased 1867:1875 arg1 complexity 1877:1886 CONCLUSIONS/INTERPRETATION Increased complexity 1840:1886 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures 1840:1916 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 8 8 theme blood 2004:2008 arg1 levels 2018:2023 blood glucose levels 2004:2023 blood glucose levels 2004:2023 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 5 9 dep illness 803:809 arg1 risk 822:825 increased risk 812:825 increased risk for development of type 2 diabetes 812:860 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 10 theme increased 1714:1722 arg1 branching 1724:1732 increased branching 1714:1732 increased branching 1714:1732 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 9 11 theme complications 2163:2175 arg1 prevention 2132:2141 prevention 2132:2141 prevention of diabetes and its complications 2132:2175 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 8 12 theme N-glycan 1898:1905 arg1 structures 1907:1916 plasma N-glycan structures 1891:1916 plasma N-glycan structures 1891:1916 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 2 13 theme prognostic 291:300 arg1 biomarkers 302:311 promising diagnostic and prognostic biomarkers 266:311 promising diagnostic and prognostic biomarkers 266:311 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 5 14 theme diabetes 853:860 arg1 development 831:841 development 831:841 development of type 2 diabetes 831:860 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 5 15 with individuals 744:754 arg1 hyperglycaemia 772:785 registered hyperglycaemia 761:785 registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) 761:861 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 16 theme diabetes 1575:1582 arg1 individuals 1631:1641 individuals 1631:1641 individuals with elevated HbA1c (ORCADES and SABRE populations) 1631:1693 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 16 theme diabetes 1575:1582 arg1 RESULTS 1411:1417 RESULTS 1411:1417 RESULTS 1411:1417 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 16 theme diabetes 1575:1582 arg1 cases 1559:1563 incident cases 1550:1563 incident cases of type 2 diabetes collected at baseline (FinRisk population) 1550:1625 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 5 17 theme N-linked 676:683 arg1 glycans 685:691 N-linked glycans 676:691 N-linked glycans from plasma proteins 676:712 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 2 18 theme plasma 201:206 arg1 N-glycome 208:216 human plasma N-glycome 195:216 human plasma N-glycome 195:216 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 7 19 theme type 1568:1571 arg1 diabetes 1575:1582 type 2 diabetes 1568:1582 type 2 diabetes collected at baseline (FinRisk population) 1568:1625 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 20 theme Disease 1196:1202 arg1 FinRisk 1096:1102 FinRisk 1096:1102 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls) 1096:1178 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 20 theme Disease 1196:1202 arg1 Study 1204:1208 Orkney Complex Disease Study 1181:1208 Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) 1181:1284 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 7 21 dep HbA1c 1657:1661 arg1 ORCADES 1664:1670 ORCADES 1664:1670 ORCADES 1664:1670 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 21 dep HbA1c 1657:1661 arg1 populations 1682:1692 SABRE populations 1676:1692 SABRE populations 1676:1692 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 22 theme Orkney 1181:1186 arg1 FinRisk 1096:1102 FinRisk 1096:1102 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls) 1096:1178 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 22 theme Orkney 1181:1186 arg1 Study 1204:1208 Orkney Complex Disease Study 1181:1208 Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) 1181:1284 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 23 from studies 1334:1340 arg1 individuals 1079:1089 individuals 1079:1089 individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls) 1079:1408 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 5 24 theme type 846:849 arg1 diabetes 853:860 type 2 diabetes 846:860 type 2 diabetes 846:860 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 0 25 theme type 72:75 arg1 diabetes 79:86 type 2 diabetes 72:86 type 2 diabetes 72:86 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 6 26 with [47.5 mmol/mol 1377:1390 arg1 % 1375:1375 HbA1c > 6.5% 1364:1375 HbA1c > 6.5% 1364:1375 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 7 27 gly sialylation 1755:1765 arg1 N-glycans 1785:1793 plasma protein N-glycans 1770:1793 plasma protein N-glycans 1770:1793 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 0 28 theme diabetes 79:86 arg1 risk 64:67 higher risk 57:67 higher risk of type 2 diabetes 57:86 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 7 29 theme elevated 1648:1655 arg1 HbA1c 1657:1661 elevated HbA1c 1648:1661 elevated HbA1c (ORCADES and SABRE populations) 1648:1693 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 30 theme AcuteInflammation 1506:1522 arg1 populations 1536:1546 AcuteInflammation Replication populations 1506:1546 AcuteInflammation Replication populations 1506:1546 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 4 31 theme study 485:489 arg1 aim 473:475 The aim 469:475 The aim of this study 469:489 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 8 32 theme type 1963:1966 arg1 diabetes 1970:1977 type 2 diabetes 1963:1977 type 2 diabetes 1963:1977 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 3 33 theme type 441:444 arg1 diabetes 448:455 type 2 diabetes 441:455 type 2 diabetes 441:455 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 0 34 theme Increased 0:8 arg1 complexity 27:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 7 35 dep RESULTS 1411:1417 arg1 Individuals 1419:1429 Individuals 1419:1429 Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations) 1419:1547 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 35 dep RESULTS 1411:1417 arg1 all 1695:1697 all 1695:1697 all 1695:1697 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 3 36 theme hexosamine 389:398 arg1 pathway 400:406 the hexosamine pathway 385:406 the hexosamine pathway 385:406 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 6 37 theme cohort 1327:1332 arg1 studies 1334:1340 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies 1096:1340 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls) 1096:1408 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 0 38 theme N-glycome 17:25 arg1 complexity 27:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 6 39 dep individuals 1347:1357 arg1 [47.5 mmol/mol 1377:1390 [47.5 mmol/mol 1377:1390 [47.5 mmol/mol 1377:1390 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 1 40 theme Better 105:110 arg1 understanding 112:124 AIMS/HYPOTHESIS Better understanding 89:124 AIMS/HYPOTHESIS Better understanding of type 2 diabetes 89:143 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 40 theme Better 105:110 arg1 need 178:181 a pressing need 167:181 a pressing need 167:181 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 40 theme Better 105:110 arg1 prevention 153:162 its prevention 149:162 its prevention 149:162 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 7 41 theme FinRisk 1607:1613 arg1 baseline 1597:1604 baseline 1597:1604 baseline (FinRisk population) 1597:1625 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 41 theme FinRisk 1607:1613 arg1 population 1615:1624 FinRisk population 1607:1624 FinRisk population 1607:1624 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 3 42 theme glycome 430:436 arg1 studies 412:418 studies 412:418 studies of plasma glycome in type 2 diabetes 412:455 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 3 42 theme glycome 430:436 arg1 scarce 461:466 scarce 461:466 scarce 461:466 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 6 43 theme HbA1c 1364:1368 arg1 % 1375:1375 HbA1c > 6.5% 1364:1375 HbA1c > 6.5% 1364:1375 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 9 44 from improvement 2117:2127 arg1 prevention 2132:2141 prevention 2132:2141 prevention of diabetes and its complications 2132:2175 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 4 45 theme protein 523:529 arg1 N-glycome 531:539 plasma protein N-glycome 516:539 plasma protein N-glycome 516:539 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 7 46 theme diabetes 1455:1462 arg1 development 1471:1481 diabetes type 2 development 1455:1481 diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations) 1455:1547 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 47 theme type 1126:1129 arg1 diabetes 1133:1140 type 2 diabetes 1126:1140 type 2 diabetes 1126:1140 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 5 48 with individuals 867:877 arg1 hyperglycaemia 772:785 registered hyperglycaemia 761:785 registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) 761:861 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 49 theme N-glycans 1785:1793 arg1 branching 1724:1732 increased branching 1714:1732 increased branching 1714:1732 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 49 theme N-glycans 1785:1793 arg1 sialylation 1755:1765 sialylation 1755:1765 sialylation 1755:1765 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 49 theme N-glycans 1785:1793 arg1 galactosylation 1735:1749 galactosylation 1735:1749 galactosylation 1735:1749 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 4 50 theme developing 594:603 arg1 diabetes 612:619 developing type 2 diabetes 594:619 developing type 2 diabetes 594:619 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 9 51 theme potential 2090:2098 arg1 approach 2104:2111 a potential new approach 2088:2111 a potential new approach for improvement in prevention of diabetes and its complications 2088:2175 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 6 52 with [47.5 mmol/mol 1253:1266 arg1 % 1251:1251 HbA1c > 6.5% 1240:1251 HbA1c > 6.5% 1240:1251 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 5 53 theme increased 812:820 arg1 risk 822:825 increased risk 812:825 increased risk for development of type 2 diabetes 812:860 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 54 theme plasma 1770:1775 arg1 N-glycans 1785:1793 plasma protein N-glycans 1770:1793 plasma protein N-glycans 1770:1793 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 5 55 theme chromatographic 638:652 arg1 approach 654:661 a chromatographic approach 636:661 a chromatographic approach 636:661 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 9 56 theme diabetes 2146:2153 arg1 prevention 2132:2141 prevention 2132:2141 prevention of diabetes and its complications 2132:2175 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 7 57 with Individuals 1419:1429 arg1 risk 1446:1449 increased risk 1436:1449 increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations) 1436:1547 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 58 theme FinRisk 1096:1102 arg1 studies 1334:1340 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies 1096:1340 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls) 1096:1408 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 8 59 theme CONCLUSIONS/INTERPRETATION 1840:1865 arg1 complexity 1877:1886 CONCLUSIONS/INTERPRETATION Increased complexity 1840:1886 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures 1840:1916 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 6 60 theme HbA1c 1240:1244 arg1 % 1251:1251 HbA1c > 6.5% 1240:1251 HbA1c > 6.5% 1240:1251 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 2 61 theme promising 266:274 arg1 biomarkers 302:311 promising diagnostic and prognostic biomarkers 266:311 promising diagnostic and prognostic biomarkers 266:311 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 5 62 theme critical 794:801 arg1 illness 803:809 critical illness 794:809 critical illness (increased risk for development of type 2 diabetes) 794:861 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 8 63 theme plasma 1891:1896 arg1 structures 1907:1916 plasma N-glycan structures 1891:1916 plasma N-glycan structures 1891:1916 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 1 64 theme pressing 169:176 arg1 understanding 112:124 AIMS/HYPOTHESIS Better understanding 89:124 AIMS/HYPOTHESIS Better understanding of type 2 diabetes 89:143 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 64 theme pressing 169:176 arg1 need 178:181 a pressing need 167:181 a pressing need 167:181 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 64 theme pressing 169:176 arg1 prevention 153:162 its prevention 149:162 its prevention 149:162 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 5 65 theme plasma 698:703 arg1 proteins 705:712 plasma proteins 698:712 plasma proteins 698:712 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 8 66 theme levels 2018:2023 arg1 diabetes 1970:1977 type 2 diabetes 1963:1977 type 2 diabetes 1963:1977 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 8 66 theme levels 2018:2023 arg1 regulation 1990:1999 poorer regulation 1983:1999 poorer regulation of blood glucose levels 1983:2023 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 6 67 theme diabetes 1133:1140 arg1 cases 1117:1121 37 incident cases 1105:1121 37 incident cases of type 2 diabetes collected at baseline 1105:1162 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 8 68 theme higher 1937:1942 arg1 risk 1944:1947 higher risk 1937:1947 higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels 1937:2023 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 8 69 theme structures 1907:1916 arg1 complexity 1877:1886 CONCLUSIONS/INTERPRETATION Increased complexity 1840:1886 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures 1840:1916 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 7 70 with individuals 1631:1641 arg1 HbA1c 1657:1661 elevated HbA1c 1648:1661 elevated HbA1c (ORCADES and SABRE populations) 1648:1693 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 5 71 dep Replication 998:1008 arg1 cases 1014:1018 52 cases 1011:1018 52 cases 1011:1018 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 5 71 dep Replication 998:1008 arg1 controls 1026:1033 14 controls 1023:1033 14 controls 1023:1033 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 1 72 theme type 129:132 arg1 diabetes 136:143 type 2 diabetes 129:143 type 2 diabetes 129:143 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 7 73 theme incident 1550:1557 arg1 cases 1559:1563 incident cases 1550:1563 incident cases of type 2 diabetes collected at baseline (FinRisk population) 1550:1625 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 74 theme Complex 1188:1194 arg1 FinRisk 1096:1102 FinRisk 1096:1102 FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls) 1096:1178 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 74 theme Complex 1188:1194 arg1 Study 1204:1208 Orkney Complex Disease Study 1181:1208 Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) 1181:1284 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 5 75 theme registered 761:770 arg1 hyperglycaemia 772:785 registered hyperglycaemia 761:785 registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) 761:861 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 2 76 theme many 238:241 arg1 diseases 243:250 many diseases 238:250 many diseases 238:250 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 1 77 theme diabetes 136:143 arg1 understanding 112:124 AIMS/HYPOTHESIS Better understanding 89:124 AIMS/HYPOTHESIS Better understanding of type 2 diabetes 89:143 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 77 theme diabetes 136:143 arg1 need 178:181 a pressing need 167:181 a pressing need 167:181 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 77 theme diabetes 136:143 arg1 prevention 153:162 its prevention 149:162 its prevention 149:162 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 6 78 dep Study 1204:1208 arg1 ORCADES 1211:1217 ORCADES 1211:1217 ORCADES 1211:1217 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 7 79 with cases 1559:1563 arg1 HbA1c 1657:1661 elevated HbA1c 1648:1661 elevated HbA1c (ORCADES and SABRE populations) 1648:1693 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 5 80 theme same 916:919 arg1 condition 921:929 the same condition 912:929 the same condition 912:929 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 81 theme SABRE 1676:1680 arg1 populations 1682:1692 SABRE populations 1676:1692 SABRE populations 1676:1692 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 82 theme Replication 1524:1534 arg1 populations 1536:1546 AcuteInflammation Replication populations 1506:1546 AcuteInflammation Replication populations 1506:1546 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 3 83 theme glucose 328:334 arg1 metabolism 336:345 glucose metabolism 328:345 glucose metabolism 328:345 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 4 84 theme plasma 516:521 arg1 N-glycome 531:539 plasma protein N-glycome 516:539 plasma protein N-glycome 516:539 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 6 85 dep ORCADES 1211:1217 arg1 controls 1276:1283 658 controls 1272:1283 658 controls 1272:1283 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 85 dep ORCADES 1211:1217 arg1 individuals 1223:1233 94 individuals 1220:1233 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] 1220:1267 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 8 86 theme poorer 1983:1988 arg1 regulation 1990:1999 poorer regulation 1983:1999 poorer regulation of blood glucose levels 1983:2023 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 6 87 dep studies 1334:1340 arg1 individuals 1347:1357 307 individuals 1343:1357 307 individuals with HbA1c > 6.5% [47.5 mmol/mol] 1343:1391 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 87 dep studies 1334:1340 arg1 controls 1400:1407 307 controls 1396:1407 307 controls 1396:1407 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 5 88 theme AcuteInflammation 980:996 arg1 Replication 998:1008 AcuteInflammation Replication 980:1008 AcuteInflammation Replication (52 cases vs 14 controls) 980:1034 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 5 89 dep AcuteInflammation 932:948 arg1 cases 954:958 59 cases 951:958 59 cases 951:958 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 5 89 dep AcuteInflammation 932:948 arg1 controls 966:973 49 controls 963:973 49 controls 963:973 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 5 89 dep AcuteInflammation 932:948 arg1 populations 1036:1046 populations 1036:1046 populations 1036:1046 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 0 90 theme plasma 10:15 arg1 complexity 27:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity 0:36 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 3 91 from studies 412:418 arg1 diabetes 448:455 type 2 diabetes 441:455 type 2 diabetes 441:455 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 1 92 theme AIMS/HYPOTHESIS 89:103 arg1 understanding 112:124 AIMS/HYPOTHESIS Better understanding 89:124 AIMS/HYPOTHESIS Better understanding of type 2 diabetes 89:143 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 92 theme AIMS/HYPOTHESIS 89:103 arg1 need 178:181 a pressing need 167:181 a pressing need 167:181 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 1 92 theme AIMS/HYPOTHESIS 89:103 arg1 prevention 153:162 its prevention 149:162 its prevention 149:162 AIMS/HYPOTHESIS Better understanding of type 2 diabetes and its prevention is a pressing need. 28905229 6 93 dep FinRisk 1096:1102 arg1 controls 1170:1177 37 controls 1167:1177 37 controls 1167:1177 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 93 dep FinRisk 1096:1102 arg1 cases 1117:1121 37 incident cases 1105:1121 37 incident cases of type 2 diabetes collected at baseline 1105:1162 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 0 94 theme higher 57:62 arg1 risk 64:67 higher risk 57:67 higher risk of type 2 diabetes 57:86 Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes. 28905229 3 95 theme plasma 423:428 arg1 glycome 430:436 plasma glycome 423:436 plasma glycome 423:436 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28905229 2 96 theme human 195:199 arg1 N-glycome 208:216 human plasma N-glycome 195:216 human plasma N-glycome 195:216 Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic and prognostic biomarkers. 28905229 7 97 theme similar 1821:1827 arg1 magnitude 1829:1837 similar magnitude 1821:1837 similar magnitude 1821:1837 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 98 theme type 1464:1467 arg1 development 1471:1481 diabetes type 2 development 1455:1481 diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations) 1455:1547 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 4 99 theme type 605:608 arg1 diabetes 612:619 developing type 2 diabetes 594:619 developing type 2 diabetes 594:619 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 5 100 link N-linked 676:683 arg1 glycans 685:691 N-linked glycans 676:691 N-linked glycans from plasma proteins 676:712 METHODS Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. 28905229 7 101 dep development 1471:1481 arg1 populations 1536:1546 AcuteInflammation Replication populations 1506:1546 AcuteInflammation Replication populations 1506:1546 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 7 101 dep development 1471:1481 arg1 AcuteInflammation 1484:1500 AcuteInflammation 1484:1500 AcuteInflammation 1484:1500 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 6 102 dep individuals 1223:1233 arg1 [47.5 mmol/mol 1253:1266 [47.5 mmol/mol 1253:1266 [47.5 mmol/mol 1253:1266 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 6 103 theme incident 1108:1115 arg1 cases 1117:1121 37 incident cases 1105:1121 37 incident cases of type 2 diabetes collected at baseline 1105:1162 N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 6.5% [47.5 mmol/mol] vs 307 controls). 28905229 4 104 theme diabetes 612:619 arg1 risk 586:589 greater risk 578:589 greater risk of developing type 2 diabetes 578:619 The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes. 28905229 9 105 theme new 2100:2102 arg1 approach 2104:2111 a potential new approach 2088:2111 a potential new approach for improvement in prevention of diabetes and its complications 2088:2175 Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications. 28905229 8 106 theme glucose 2010:2016 arg1 levels 2018:2023 blood glucose levels 2004:2023 blood glucose levels 2004:2023 CONCLUSIONS/INTERPRETATION Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. 28905229 7 107 with RESULTS 1411:1417 arg1 HbA1c 1657:1661 elevated HbA1c 1648:1661 elevated HbA1c (ORCADES and SABRE populations) 1648:1693 RESULTS Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA1c (ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude. 28905229 3 108 from Variations 314:323 arg1 metabolism 336:345 glucose metabolism 328:345 glucose metabolism 328:345 Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. 28074929 5 0 contain containing 822:831 arg2 domains 837:843 the domains 833:843 the domains 833:843 From 153 species, 4491 sequences containing the domains were retrieved, based on which we analyzed distribution of domains among eukaryotic species. 28074929 5 0 contain containing 822:831 arg1 sequences 812:820 4491 sequences 807:820 4491 sequences containing the domains 807:843 From 153 species, 4491 sequences containing the domains were retrieved, based on which we analyzed distribution of domains among eukaryotic species. 28074929 1 1 theme Protein 148:154 arg1 PNG 173:175 PNG 173:175 PNG 173:175 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 1 1 theme Protein 148:154 arg1 N-glycosylation 156:170 Protein N-glycosylation 148:170 Protein N-glycosylation (PNG) 148:176 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 7 2 theme plant-specific 1288:1301 arg1 structure 1307:1315 plant-specific Lea structure 1288:1315 plant-specific Lea structure 1288:1315 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 6 3 from domains 942:948 arg1 GnTV 953:956 GnTV 953:956 GnTV 953:956 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 3 4 theme eukaryotic 483:492 arg1 structures 511:520 eukaryotic protein N-glycan structures 483:520 eukaryotic protein N-glycan structures 483:520 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 0 5 theme N-glycan 98:105 arg1 structures 107:116 protein N-glycan structures 90:116 protein N-glycan structures 90:116 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 0 6 from Evolution 0:8 arg1 apparatus 54:62 Golgi apparatus 48:62 Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi 48:145 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 7 7 theme Lea 1303:1305 arg1 structure 1307:1315 plant-specific Lea structure 1288:1315 plant-specific Lea structure 1288:1315 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 6 8 theme specific 976:983 arg1 domains 996:1002 specific eukaryotic domains 976:1002 specific eukaryotic domains 976:1002 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 1 9 theme remarkable 243:252 arg1 diversity 254:262 remarkable diversity 243:262 remarkable diversity among eukaryotic species 243:287 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 0 10 theme protein 90:96 arg1 structures 107:116 protein N-glycan structures 90:116 protein N-glycan structures 90:116 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 5 11 theme domains 904:910 arg1 distribution 888:899 distribution 888:899 distribution of domains among eukaryotic species 888:935 From 153 species, 4491 sequences containing the domains were retrieved, based on which we analyzed distribution of domains among eukaryotic species. 28074929 6 12 theme eukaryotic 985:994 arg1 domains 996:1002 specific eukaryotic domains 976:1002 specific eukaryotic domains 976:1002 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 3 13 theme structures 511:520 arg1 diversity 470:478 diversity 470:478 diversity of eukaryotic protein N-glycan structures 470:520 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 4 14 theme known 647:651 arg1 genes 657:661 24 known PNG genes 644:661 24 known PNG genes 644:661 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 3 15 theme evolution 411:419 arg1 picture 390:396 a picture 388:396 a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution 388:609 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 0 16 theme structures 107:116 arg1 diversity 77:85 diversity 77:85 diversity of protein N-glycan structures in plants, animals and fungi 77:145 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 4 17 theme genes 782:786 arg1 majority 766:773 the majority 762:773 the majority of the genes 762:786 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 3 18 from apparatus 448:456 arg1 evolution 411:419 evolution 411:419 evolution 411:419 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 18 from apparatus 448:456 arg1 onset 401:405 onset 401:405 onset 401:405 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 5 19 theme eukaryotic 918:927 arg1 species 929:935 eukaryotic species 918:935 eukaryotic species 918:935 From 153 species, 4491 sequences containing the domains were retrieved, based on which we analyzed distribution of domains among eukaryotic species. 28074929 7 20 theme essential 1260:1268 arg1 enzyme 1270:1275 an essential enzyme 1257:1275 an essential enzyme in forming plant-specific Lea structure 1257:1315 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 20 theme essential 1260:1268 arg1 galactosyltransferase 1234:1254 β-1,3 galactosyltransferase 1228:1254 β-1,3 galactosyltransferase 1228:1254 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 21 attach present 1323:1329 arg2 domains 1208:1214 two domains 1204:1214 two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, 1204:1316 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 21 attach present 1323:1329 arg1 genes 1344:1348 separated genes 1334:1348 separated genes in fungi and animals 1334:1369 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 6 22 theme other 1168:1172 arg1 lineages 1185:1192 other ehkaryotic lineages 1168:1192 other ehkaryotic lineages 1168:1192 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 7 23 theme separated 1334:1342 arg1 genes 1344:1348 separated genes 1334:1348 separated genes in fungi and animals 1334:1369 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 3 24 theme onset 401:405 arg1 picture 390:396 a picture 388:396 a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution 388:609 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 7 25 from present 1323:1329 arg1 genes 1344:1348 separated genes 1334:1348 separated genes in fungi and animals 1334:1369 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 0 26 theme N-glycosylation 21:35 arg1 process 37:43 protein N-glycosylation process 13:43 protein N-glycosylation process 13:43 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 1 27 theme eukaryotic 270:279 arg1 species 281:287 eukaryotic species 270:287 eukaryotic species 270:287 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 4 28 theme single 705:710 arg1 clan 712:715 a single clan 703:715 a single clan 703:715 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 0 29 theme protein 13:19 arg1 process 37:43 protein N-glycosylation process 13:43 protein N-glycosylation process 13:43 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 6 30 theme PNG 1074:1076 arg1 reactions 1078:1086 certain unique PNG reactions 1059:1086 certain unique PNG reactions 1059:1086 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 3 31 theme PNG 424:426 arg1 components 428:437 PNG components 424:437 PNG components in Golgi apparatus 424:456 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 32 theme Golgi 442:446 arg1 apparatus 448:456 Golgi apparatus 442:456 Golgi apparatus 442:456 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 7 33 located present 1323:1329 arg2 domains 1208:1214 two domains 1204:1214 two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, 1204:1316 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 33 located present 1323:1329 arg1 genes 1344:1348 separated genes 1334:1348 separated genes in fungi and animals 1334:1369 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 6 34 theme certain 1059:1065 arg1 reactions 1078:1086 certain unique PNG reactions 1059:1086 certain unique PNG reactions 1059:1086 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 0 35 from diversity 77:85 arg1 animals 129:135 animals 129:135 animals 129:135 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 0 35 from diversity 77:85 arg1 plants 121:126 plants 121:126 plants 121:126 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 0 35 from diversity 77:85 arg1 fungi 141:145 fungi 141:145 fungi 141:145 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 1 36 theme protein 193:199 arg1 folding 201:207 protein folding 193:207 protein folding 193:207 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 0 37 theme process 37:43 arg1 Evolution 0:8 Evolution 0:8 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 0:146 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 5 38 theme 4491 807:810 arg1 sequences 812:820 4491 sequences 807:820 4491 sequences containing the domains 807:843 From 153 species, 4491 sequences containing the domains were retrieved, based on which we analyzed distribution of domains among eukaryotic species. 28074929 6 39 dep distribute 1022:1031 arg1 only 1037:1040 only 1037:1040 only 1037:1040 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 3 40 theme components 428:437 arg1 evolution 411:419 evolution 411:419 evolution 411:419 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 40 theme components 428:437 arg1 onset 401:405 onset 401:405 onset 401:405 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 41 theme domain 554:559 arg1 emergence 561:569 domain emergence 554:569 domain emergence 554:569 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 42 from components 428:437 arg1 apparatus 448:456 Golgi apparatus 442:456 Golgi apparatus 442:456 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 6 43 theme unique 1067:1072 arg1 reactions 1078:1086 certain unique PNG reactions 1059:1086 certain unique PNG reactions 1059:1086 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 0 44 theme Golgi 48:52 arg1 apparatus 54:62 Golgi apparatus 48:62 Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi 48:145 Evolution of protein N-glycosylation process in Golgi apparatus which shapes diversity of protein N-glycan structures in plants, animals and fungi. 28074929 6 45 theme ehkaryotic 1174:1183 arg1 lineages 1185:1192 other ehkaryotic lineages 1168:1192 other ehkaryotic lineages 1168:1192 Two domains in GnTV are restricted to specific eukaryotic domains, while 10 domains distribute not only in species where certain unique PNG reactions occur and thus genes harboring these domains are supoosed to be present, but in other ehkaryotic lineages. 28074929 2 46 theme PNG 320:322 arg1 mechanisms 324:333 unique PNG mechanisms 313:333 unique PNG mechanisms 313:333 Little is known of how unique PNG mechanisms arose and evolved in eukaryotes. 28074929 3 47 from emphasis 531:538 arg1 roles 543:547 roles 543:547 roles that domain emergence and combination played on PNG evolution 543:609 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 4 48 theme evolutionary 738:749 arg1 source 751:756 a single evolutionary source 729:756 a single evolutionary source for the majority of the genes 729:786 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 3 49 theme PNG 597:599 arg1 evolution 601:609 PNG evolution 597:609 PNG evolution 597:609 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 2 50 theme unique 313:318 arg1 mechanisms 324:333 unique PNG mechanisms 313:333 unique PNG mechanisms 313:333 Little is known of how unique PNG mechanisms arose and evolved in eukaryotes. 28074929 7 51 theme domain 1412:1417 arg1 shuffling 1419:1427 domain shuffling 1412:1427 domain shuffling 1412:1427 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 4 52 theme single 731:736 arg1 source 751:756 a single evolutionary source 729:756 a single evolutionary source for the majority of the genes 729:786 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 3 53 from onset 401:405 arg1 apparatus 448:456 Golgi apparatus 442:456 Golgi apparatus 442:456 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 4 54 theme PNG 653:655 arg1 genes 657:661 24 known PNG genes 644:661 24 known PNG genes 644:661 23 domains were identified from 24 known PNG genes, most of which could be classified into a single clan, indicating a single evolutionary source for the majority of the genes. 28074929 1 55 theme enzymatic 213:221 arg1 activities 223:232 enzymatic activities 213:232 enzymatic activities 213:232 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 7 56 theme shuffling 1419:1427 arg1 emergence 1387:1395 its emergence 1383:1395 its emergence 1383:1395 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 56 theme shuffling 1419:1427 arg1 result 1402:1407 a result 1400:1407 a result of domain shuffling 1400:1427 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 1 57 contain has 239:241 arg2 diversity 254:262 remarkable diversity 243:262 remarkable diversity among eukaryotic species 243:287 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 1 57 contain has 239:241 arg1 PNG 173:175 PNG 173:175 PNG 173:175 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 1 57 contain has 239:241 arg1 N-glycosylation 156:170 Protein N-glycosylation 148:170 Protein N-glycosylation (PNG) 148:176 Protein N-glycosylation (PNG) is crucial for protein folding and enzymatic activities, and has remarkable diversity among eukaryotic species. 28074929 3 58 theme protein 494:500 arg1 structures 511:520 eukaryotic protein N-glycan structures 483:520 eukaryotic protein N-glycan structures 483:520 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 7 59 from genes 1344:1348 arg1 fungi 1353:1357 fungi 1353:1357 fungi 1353:1357 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 59 from genes 1344:1348 arg1 present 1323:1329 present 1323:1329 present 1323:1329 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 7 59 from genes 1344:1348 arg1 animals 1363:1369 animals 1363:1369 animals 1363:1369 Notably, two domains harbored by β-1,3 galactosyltransferase, an essential enzyme in forming plant-specific Lea structure, were present in separated genes in fungi and animals, suggesting its emergence as a result of domain shuffling. 28074929 3 60 from evolution 411:419 arg1 apparatus 448:456 Golgi apparatus 442:456 Golgi apparatus 442:456 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 28074929 3 61 theme N-glycan 502:509 arg1 structures 511:520 eukaryotic protein N-glycan structures 483:520 eukaryotic protein N-glycan structures 483:520 Here we demonstrate a picture of onset and evolution of PNG components in Golgi apparatus that shaped diversity of eukaryotic protein N-glycan structures, with an emphasis on roles that domain emergence and combination played on PNG evolution. 25488985 1 0 from generation 351:360 arg1 insects 394:400 insects 394:400 insects 394:400 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 3 1 theme FDL 770:772 arg1 homolog 774:780 the Apis mellifera FDL homolog 751:780 the Apis mellifera FDL homolog 751:780 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 7 2 from exohexosaminidases 1835:1852 arg1 insects 1857:1863 insects 1857:1863 insects 1857:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 3 3 from melanogaster 696:707 arg1 enzyme 673:678 the already-known FDL enzyme 651:678 the already-known FDL enzyme from Drosophila melanogaster 651:707 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 3 4 theme Apis 755:758 arg1 mellifera 760:768 the Apis mellifera 751:768 the Apis mellifera FDL homolog 751:780 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 6 5 theme Caenorhabditis 1519:1532 arg1 strains 1534:1540 wild-type and mutant Caenorhabditis strains 1498:1540 wild-type and mutant Caenorhabditis strains 1498:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 7 6 from insects 1857:1863 arg1 similarity 1785:1794 the high structural similarity 1765:1794 the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1765:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 8 7 theme important 1986:1994 arg1 family 1979:1984 the GH20 hexosaminidase family 1955:1984 the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1955:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 7 8 theme compatible 1749:1758 arg1 FDL 1744:1746 Drosophila FDL 1733:1746 Drosophila FDL 1733:1746 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 6 9 theme N-glycans 1608:1616 arg1 definition 1567:1576 isomeric definition 1558:1576 isomeric definition of paucimannosidic and hybrid N-glycans 1558:1616 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 7 10 theme FDL 1744:1746 arg1 mutants 1722:1728 two site-directed mutants 1704:1728 two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1704:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 4 11 theme recombinant 974:984 arg1 enzymes 1018:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 8 12 from variety 1909:1915 arg1 family 1979:1984 the GH20 hexosaminidase family 1955:1984 the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1955:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 6 13 theme Recombinant 1397:1407 arg1 FDL 1409:1411 Recombinant FDL 1397:1411 Recombinant FDL 1397:1411 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 6 14 theme mutant 1512:1517 arg1 strains 1534:1540 wild-type and mutant Caenorhabditis strains 1498:1540 wild-type and mutant Caenorhabditis strains 1498:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 2 15 theme relevant 605:612 arg1 glycosyltransferases 614:633 various relevant glycosyltransferases 597:633 various relevant glycosyltransferases 597:633 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 7 16 from differences 1649:1659 arg1 activity 1664:1671 activity 1664:1671 activity 1664:1671 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 7 16 from differences 1649:1659 arg1 specificity 1677:1687 specificity 1677:1687 specificity 1677:1687 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 1 17 theme Fused 152:156 arg1 glycosidases 228:239 the most recently genetically defined glycosidases 190:239 the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates 190:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 1 17 theme Fused 152:156 arg1 hexosaminidases 170:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 7 18 theme site-directed 1708:1720 arg1 mutants 1722:1728 two site-directed mutants 1704:1728 two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1704:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 7 19 from similarity 1785:1794 arg1 insects 1857:1863 insects 1857:1863 insects 1857:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 5 20 theme other 1341:1345 arg1 GalNAc 1356:1361 other terminal GalNAc and N-acetylglucosamine residues 1341:1394 GalNAc 1356:1361 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 1 21 from biosynthesis 257:268 arg1 invertebrates 286:298 invertebrates 286:298 invertebrates 286:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 3 22 theme FDL 669:671 arg1 enzyme 673:678 the already-known FDL enzyme 651:678 the already-known FDL enzyme from Drosophila melanogaster 651:707 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 8 23 from family 1979:1984 arg1 aspects 1944:1950 structural and function aspects 1920:1950 structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1920:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 8 23 from family 1979:1984 arg1 variety 1909:1915 the variety 1905:1915 the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1905:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 6 24 theme mixtures 1459:1466 arg1 analysis 1439:1446 the analysis 1435:1446 the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains 1435:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 8 25 theme structural 1920:1929 arg1 aspects 1944:1950 structural and function aspects 1920:1950 structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1920:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 7 26 theme exohexosaminidases 1835:1852 arg1 similarity 1785:1794 the high structural similarity 1765:1794 the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1765:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 2 27 theme substrates 522:531 arg1 utilization 474:484 the utilization 470:484 the utilization of different artificial and natural substrates 470:531 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 5 28 dep specific 1237:1244 arg1 enzymes 1218:1224 the invertebrate FDL(-like) enzymes 1190:1224 the invertebrate FDL(-like) enzymes 1190:1224 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 28 dep specific 1237:1244 arg1 remove 1334:1339 remove 1334:1339 remove other terminal GalNAc and N-acetylglucosamine residues 1334:1394 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 28 dep specific 1237:1244 arg1 specific 1237:1244 specific 1237:1244 specific 1237:1244 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 29 theme extreme 1310:1316 arg1 conditions 1318:1327 extreme conditions 1310:1327 extreme conditions 1310:1327 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 1 30 theme N-glycans 381:389 arg1 generation 351:360 the generation 347:360 the generation of paucimannosidic N-glycans in insects 347:400 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 8 31 theme function 1935:1942 arg1 aspects 1944:1950 structural and function aspects 1920:1950 structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1920:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 7 32 theme N-glycan 1816:1823 arg1 exohexosaminidases 1835:1852 chitinolytic and N-glycan degrading exohexosaminidases 1799:1852 exohexosaminidases 1835:1852 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 4 33 theme elegans 1001:1007 arg1 enzymes 1018:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 0 34 theme invertebrate 102:113 arg1 hexosaminidases 135:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 5 35 theme natural 1158:1164 arg1 N-glycans 1166:1174 natural N-glycans 1158:1174 natural N-glycans 1158:1174 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 7 36 theme chitinolytic 1799:1810 arg1 exohexosaminidases 1835:1852 chitinolytic and N-glycan degrading exohexosaminidases 1799:1852 exohexosaminidases 1835:1852 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 2 37 theme purified 542:549 arg1 compounds 559:567 purified, native compounds 542:567 purified, native compounds 542:567 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 4 38 theme forms 823:827 arg1 properties 797:806 The enzymatic properties 783:806 The enzymatic properties 783:806 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 0 39 theme processing 124:133 arg1 hexosaminidases 135:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 6 40 from analysis 1439:1446 arg1 useful 1425:1430 useful 1425:1430 useful 1425:1430 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 8 41 theme GH20 1959:1962 arg1 family 1979:1984 the GH20 hexosaminidase family 1955:1984 the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1955:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 0 42 from differences 32:42 arg1 specificity 61:71 the substrate specificity 47:71 the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases 47:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 0 43 theme Enzymatic 0:8 arg1 properties 10:19 Enzymatic properties 0:19 Enzymatic properties 0:19 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 7 44 theme high 1769:1772 arg1 similarity 1785:1794 the high structural similarity 1765:1794 the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1765:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 5 45 theme N-acetylglucosamine 1367:1385 arg1 residues 1387:1394 other terminal GalNAc and N-acetylglucosamine residues 1341:1394 residues 1387:1394 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 2 46 theme FDL 447:449 arg1 hexosaminidases 451:465 FDL hexosaminidases 447:465 FDL hexosaminidases 447:465 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 4 47 theme enzymes 865:871 arg1 forms 823:827 the soluble forms 811:827 the soluble forms of the affinity-purified insect FDL enzymes 811:871 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 5 48 theme FDL 1207:1209 arg1 remove 1334:1339 remove 1334:1339 remove other terminal GalNAc and N-acetylglucosamine residues 1334:1394 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 48 theme FDL 1207:1209 arg1 enzymes 1218:1224 the invertebrate FDL(-like) enzymes 1190:1224 the invertebrate FDL(-like) enzymes 1190:1224 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 48 theme FDL 1207:1209 arg1 specific 1237:1244 specific 1237:1244 specific 1237:1244 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 4 49 theme Caenorhabditis 1074:1087 arg1 HEX-4 1104:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 6 50 theme N-glycans 1471:1479 arg1 mixtures 1459:1466 complex mixtures 1451:1466 complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains 1451:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 6 51 theme paucimannosidic 1581:1595 arg1 N-glycans 1608:1616 paucimannosidic and hybrid N-glycans 1581:1616 N-glycans 1608:1616 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 8 52 theme hexosaminidase 1964:1977 arg1 family 1979:1984 the GH20 hexosaminidase family 1955:1984 the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1955:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 4 53 theme insect 854:859 arg1 enzymes 865:871 the affinity-purified insect FDL enzymes 832:871 the affinity-purified insect FDL enzymes 832:871 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 2 54 theme different 489:497 arg1 substrates 522:531 different artificial and natural substrates 489:531 different artificial and natural substrates 489:531 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 6 55 theme complex 1451:1457 arg1 mixtures 1459:1466 complex mixtures 1451:1466 complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains 1451:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 4 56 theme insect 902:907 arg1 cells 909:913 insect cells 902:913 insect cells 902:913 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 3 57 theme mellifera 760:768 arg1 homolog 774:780 the Apis mellifera FDL homolog 751:780 the Apis mellifera FDL homolog 751:780 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 8 58 theme glycoconjugates 2036:2050 arg1 biosynthesis 2020:2031 biosynthesis 2020:2031 biosynthesis 2020:2031 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 8 58 theme glycoconjugates 2036:2050 arg1 catabolism 2005:2014 catabolism 2005:2014 catabolism 2005:2014 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 1 59 theme defined 220:226 arg1 glycosidases 228:239 the most recently genetically defined glycosidases 190:239 the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates 190:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 1 59 theme defined 220:226 arg1 hexosaminidases 170:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 7 60 theme Drosophila 1733:1742 arg1 FDL 1744:1746 Drosophila FDL 1733:1746 Drosophila FDL 1733:1746 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 5 61 theme substrates 1136:1145 arg1 range 1127:1131 a range 1125:1131 a range of substrates, including natural N-glycans 1125:1174 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 61 theme substrates 1136:1145 arg1 N-glycans 1166:1174 natural N-glycans 1158:1174 natural N-glycans 1158:1174 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 6 62 theme hybrid 1601:1606 arg1 N-glycans 1608:1616 paucimannosidic and hybrid N-glycans 1581:1616 N-glycans 1608:1616 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 4 63 theme distinct 965:972 arg1 enzymes 1018:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 2 64 theme various 597:603 arg1 glycosyltransferases 614:633 various relevant glycosyltransferases 597:633 various relevant glycosyltransferases 597:633 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 8 65 from aspects 1944:1950 arg1 family 1979:1984 the GH20 hexosaminidase family 1955:1984 the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1955:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 3 66 theme already-known 655:667 arg1 enzyme 673:678 the already-known FDL enzyme 651:678 the already-known FDL enzyme from Drosophila melanogaster 651:707 In addition to the already-known FDL enzyme from Drosophila melanogaster, we now have identified and characterized the Apis mellifera FDL homolog. 25488985 1 67 theme lobes 158:162 arg1 glycosidases 228:239 the most recently genetically defined glycosidases 190:239 the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates 190:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 1 67 theme lobes 158:162 arg1 hexosaminidases 170:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 5 68 theme terminal 1347:1354 arg1 GalNAc 1356:1361 other terminal GalNAc and N-acetylglucosamine residues 1341:1394 GalNAc 1356:1361 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 1 69 theme FDL 165:167 arg1 glycosidases 228:239 the most recently genetically defined glycosidases 190:239 the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates 190:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 1 69 theme FDL 165:167 arg1 hexosaminidases 170:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases 152:184 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 5 70 attach attached 1270:1277 arg1 α1,3-mannose 1286:1297 the α1,3-mannose 1282:1297 the α1,3-mannose 1282:1297 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 70 attach attached 1270:1277 arg2 N-acetylglucosamine 1250:1268 N-acetylglucosamine 1250:1268 N-acetylglucosamine attached to the α1,3-mannose 1250:1297 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 0 71 theme distinct 93:100 arg1 hexosaminidases 135:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 1 72 theme paucimannosidic 365:379 arg1 N-glycans 381:389 paucimannosidic N-glycans 365:389 paucimannosidic N-glycans 365:389 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 2 73 theme natural 514:520 arg1 substrates 522:531 different artificial and natural substrates 489:531 different artificial and natural substrates 489:531 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 0 74 from properties 10:19 arg1 specificity 61:71 the substrate specificity 47:71 the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases 47:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 0 75 theme N-glycan 115:122 arg1 hexosaminidases 135:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 phylogenetically distinct invertebrate N-glycan processing hexosaminidases 76:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 5 76 theme invertebrate 1194:1205 arg1 FDL 1207:1209 invertebrate FDL 1194:1209 the invertebrate FDL(-like) enzymes 1190:1224 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 7 77 theme degrading 1825:1833 arg1 exohexosaminidases 1835:1852 chitinolytic and N-glycan degrading exohexosaminidases 1799:1852 exohexosaminidases 1835:1852 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 0 78 theme hexosaminidases 135:149 arg1 specificity 61:71 the substrate specificity 47:71 the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases 47:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 6 79 from useful 1425:1430 arg1 analysis 1439:1446 the analysis 1435:1446 the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains 1435:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 8 80 theme aspects 1944:1950 arg1 aspects 1944:1950 structural and function aspects 1920:1950 structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1920:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 8 80 theme aspects 1944:1950 arg1 variety 1909:1915 the variety 1905:1915 the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes 1905:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 4 81 theme FDL-like 1009:1016 arg1 enzymes 1018:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes 944:1024 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 5 82 dep FDL 1207:1209 arg1 -like 1211:1215 -like 1211:1215 -like 1211:1215 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 4 83 theme soluble 815:821 arg1 forms 823:827 the soluble forms 811:827 the soluble forms of the affinity-purified insect FDL enzymes 811:871 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 5 84 with tests 1114:1118 arg1 range 1127:1131 a range 1125:1131 a range of substrates, including natural N-glycans 1125:1174 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 5 84 with tests 1114:1118 arg1 N-glycans 1166:1174 natural N-glycans 1158:1174 natural N-glycans 1158:1174 In tests with a range of substrates, including natural N-glycans, we show that the invertebrate FDL(-like) enzymes are highly specific for N-acetylglucosamine attached to the α1,3-mannose, but under extreme conditions also remove other terminal GalNAc and N-acetylglucosamine residues. 25488985 2 85 dep utilization 474:484 arg1 both 534:537 both 534:537 both 534:537 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 4 86 theme hexosaminidase 1089:1102 arg1 HEX-4 1104:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 8 87 from catabolism 2005:2014 arg1 eukaryotes 2055:2064 eukaryotes 2055:2064 eukaryotes 2055:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 0 88 theme subtle 25:30 arg1 differences 32:42 subtle differences 25:42 subtle differences 25:42 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 1 89 theme narrow 311:316 arg1 specificity 318:328 their narrow specificity 305:328 their narrow specificity 305:328 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25488985 2 90 dep purified 542:549 arg1 native 552:557 native 552:557 native 552:557 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 7 91 with compatible 1749:1758 arg1 similarity 1785:1794 the high structural similarity 1765:1794 the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1765:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 7 92 theme structural 1774:1783 arg1 similarity 1785:1794 the high structural similarity 1765:1794 the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects 1765:1863 Furthermore, differences in activity and specificity were shown for two site-directed mutants of Drosophila FDL, compatible with the high structural similarity of chitinolytic and N-glycan degrading exohexosaminidases in insects. 25488985 2 93 theme hexosaminidases 451:465 arg1 potential 434:442 the potential 430:442 the potential of FDL hexosaminidases 430:465 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 4 94 theme enzymatic 787:795 arg1 properties 797:806 The enzymatic properties 783:806 The enzymatic properties 783:806 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 6 95 theme wild-type 1498:1506 arg1 strains 1534:1540 wild-type and mutant Caenorhabditis strains 1498:1540 wild-type and mutant Caenorhabditis strains 1498:1540 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 2 96 theme artificial 499:508 arg1 substrates 522:531 different artificial and natural substrates 489:531 different artificial and natural substrates 489:531 In this study, we explored the potential of FDL hexosaminidases in the utilization of different artificial and natural substrates, both as purified, native compounds or generated in vitro using various relevant glycosyltransferases. 25488985 4 97 theme FDL 861:863 arg1 enzymes 865:871 the affinity-purified insect FDL enzymes 832:871 the affinity-purified insect FDL enzymes 832:871 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 0 98 theme substrate 51:59 arg1 specificity 61:71 the substrate specificity 47:71 the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases 47:149 Enzymatic properties and subtle differences in the substrate specificity of phylogenetically distinct invertebrate N-glycan processing hexosaminidases. 25488985 4 99 theme -specific 1064:1072 arg1 HEX-4 1104:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4 1030:1108 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 8 100 from biosynthesis 2020:2031 arg1 eukaryotes 2055:2064 eukaryotes 2055:2064 eukaryotes 2055:2064 Our studies are another indication for the variety of structural and function aspects in the GH20 hexosaminidase family important for both catabolism and biosynthesis of glycoconjugates in eukaryotes. 25488985 4 101 theme affinity-purified 836:852 arg1 enzymes 865:871 the affinity-purified insect FDL enzymes 832:871 the affinity-purified insect FDL enzymes 832:871 The enzymatic properties of the soluble forms of the affinity-purified insect FDL enzymes, expressed in both yeast and insect cells, were compared with those of the phylogenetically distinct recombinant Caenorhabditis elegans FDL-like enzymes and the N-acetylgalactosamine (GalNAc)-specific Caenorhabditis hexosaminidase HEX-4. 25488985 6 102 theme isomeric 1558:1565 arg1 definition 1567:1576 isomeric definition 1558:1576 isomeric definition of paucimannosidic and hybrid N-glycans 1558:1616 Recombinant FDL also proved useful in the analysis of complex mixtures of N-glycans originating from wild-type and mutant Caenorhabditis strains, thereby aiding isomeric definition of paucimannosidic and hybrid N-glycans in this organism. 25488985 1 103 theme N-glycans 273:281 arg1 biosynthesis 257:268 the biosynthesis 253:268 the biosynthesis of N-glycans in invertebrates 253:298 Fused lobes (FDL) hexosaminidases are the most recently genetically defined glycosidases involved in the biosynthesis of N-glycans in invertebrates, and their narrow specificity is essential for the generation of paucimannosidic N-glycans in insects. 25341402 0 0 from activity 161:168 arg1 roles 89:93 roles 89:93 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 7 1 theme Additional 843:852 arg1 2+ 857:858 2+ 857:858 2+ 857:858 Additional Ca(2+) was not required to measure PelA activity in vitro, but it could significantly enhance its activity and thermal stability. 25341402 7 1 theme Additional 843:852 arg1 Ca 854:855 Additional Ca 843:855 Additional Ca(2+) 843:859 Additional Ca(2+) was not required to measure PelA activity in vitro, but it could significantly enhance its activity and thermal stability. 25341402 9 2 theme residue 1268:1274 arg1 N95 1276:1278 the residue N95 1264:1278 the residue N95 1264:1278 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 0 3 from secretion 147:155 arg1 roles 89:93 roles 89:93 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 5 4 theme non-conserved 642:654 arg1 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 4 theme non-conserved 642:654 arg1 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 1 5 theme Alkaline 171:178 arg1 lyases 188:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 8 6 theme K 1133:1133 arg1 values 1137:1142 the K m values 1129:1142 the K m values 1129:1142 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 11 7 theme thermal 1589:1595 arg1 stability 1597:1605 less thermal stability 1584:1605 less thermal stability 1584:1605 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 5 8 theme N-glycosylation 666:680 arg1 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 8 theme N-glycosylation 666:680 arg1 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 2 9 theme few 312:314 arg1 fungi 316:320 a few fungi 310:320 a few fungi 310:320 They are isolated predominantly from bacteria and a few fungi. 25341402 8 10 theme mM 1110:1111 arg1 2+ 1116:1117 2+ 1116:1117 2+ 1116:1117 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 8 10 theme mM 1110:1111 arg1 Ca 1113:1114 0.1 mM Ca 1106:1114 0.1 mM Ca(2+) 1106:1118 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 11 11 from decrease 1550:1557 arg1 stability 1597:1605 less thermal stability 1584:1605 less thermal stability 1584:1605 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 11 from decrease 1550:1557 arg1 activity 1571:1578 specific activity 1562:1578 specific activity 1562:1578 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 8 12 dep increased 1047:1055 arg1 whereas 1121:1127 whereas 1121:1127 whereas 1121:1127 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 0 13 theme conserved 109:117 arg1 site 135:138 the highly conserved N-glycosylation site 98:138 the highly conserved N-glycosylation site in its secretion and activity 98:168 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 1 14 theme great 200:204 arg1 potential 218:226 great application potential 200:226 great application potential 200:226 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 11 15 contain had 1463:1465 arg2 effect 1470:1475 no effect 1467:1475 no effect 1467:1475 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 15 contain had 1463:1465 arg1 substitution 1430:1441 Amino acid substitution 1419:1441 Amino acid substitution at the residue N198 1419:1461 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 0 16 from volvacea 79:86 arg1 characteristics 12:26 Biochemical characteristics 0:26 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 5 17 gly N-glycosylation 666:680 arg2 one 620:622 one 620:622 one 620:622 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 17 gly N-glycosylation 666:680 arg2 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 17 gly N-glycosylation 666:680 arg2 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 0 18 theme site 135:138 arg1 roles 89:93 roles 89:93 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 4 19 theme full-length 459:469 arg1 polypeptide 501:511 a 321-amino-acid polypeptide 484:511 a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain 484:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 4 19 theme full-length 459:469 arg1 pelA 471:474 The full-length pelA 455:474 The full-length pelA 455:474 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 0 20 gly N-glycosylation 119:133 arg2 site 135:138 the highly conserved N-glycosylation site 98:138 the highly conserved N-glycosylation site in its secretion and activity 98:168 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 0 21 from roles 89:93 arg1 secretion 147:155 its secretion 143:155 its secretion 143:155 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 0 21 from roles 89:93 arg1 activity 161:168 activity 161:168 activity 161:168 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 11 22 theme specific 1562:1569 arg1 activity 1571:1578 specific activity 1562:1578 specific activity 1562:1578 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 3 23 theme biochemical 343:353 arg1 characteristics 355:369 the biochemical characteristics 339:369 the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 339:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 24 theme putative 526:533 arg1 peptide 553:559 a putative 18-residue signal peptide 524:559 a putative 18-residue signal peptide 524:559 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 5 25 theme conserved 624:632 arg1 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 25 theme conserved 624:632 arg1 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 3 26 theme Volvariella 433:443 arg1 volvacea 445:452 the basidiomycete Volvariella volvacea 415:452 the basidiomycete Volvariella volvacea 415:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 8 27 theme max 990:992 arg1 values 994:999 The V max values 984:999 The V max values using polygalacturonic acid as substrate 984:1040 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 7 28 theme PelA 889:892 arg1 activity 894:901 PelA activity 889:901 PelA activity 889:901 Additional Ca(2+) was not required to measure PelA activity in vitro, but it could significantly enhance its activity and thermal stability. 25341402 3 29 theme novel 376:380 arg1 PelA 405:408 a novel alkaline pectate lyase PelA 374:408 a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 374:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 30 theme 321-amino-acid 486:499 arg1 polypeptide 501:511 a 321-amino-acid polypeptide 484:511 a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain 484:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 4 30 theme 321-amino-acid 486:499 arg1 pelA 471:474 The full-length pelA 455:474 The full-length pelA 455:474 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 8 31 dep IU 1077:1078 arg1 89.96 1071:1075 89.96 1071:1075 89.96 1071:1075 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 0 32 theme Biochemical 0:10 arg1 characteristics 12:26 Biochemical characteristics 0:26 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 4 33 theme pectate 567:573 arg1 domain 600:605 a pectate lyase family 1 catalytic domain 565:605 a pectate lyase family 1 catalytic domain 565:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 9 34 theme Site-directed 1190:1202 arg1 mutagenesis 1204:1214 Site-directed mutagenesis 1190:1214 Site-directed mutagenesis 1190:1214 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 3 35 theme PelA 405:408 arg1 characteristics 355:369 the biochemical characteristics 339:369 the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 339:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 36 theme family 581:586 arg1 domain 600:605 a pectate lyase family 1 catalytic domain 565:605 a pectate lyase family 1 catalytic domain 565:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 0 37 theme alkaline 34:41 arg1 PelA 57:60 an alkaline pectate lyase PelA 31:60 an alkaline pectate lyase PelA 31:60 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 3 38 theme pectate 391:397 arg1 PelA 405:408 a novel alkaline pectate lyase PelA 374:408 a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 374:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 39 theme signal 546:551 arg1 peptide 553:559 a putative 18-residue signal peptide 524:559 a putative 18-residue signal peptide 524:559 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 11 40 theme slight 1514:1519 arg1 decrease 1550:1557 a slight (5.16 %) to modest (27.37 %) decrease 1512:1557 a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability 1512:1605 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 0 41 theme lyase 51:55 arg1 PelA 57:60 an alkaline pectate lyase PelA 31:60 an alkaline pectate lyase PelA 31:60 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 6 42 theme optimal 759:765 arg1 activity 767:774 optimal activity 759:774 optimal activity 759:774 The enzyme showed optimal activity at 60 °C and pH 10, although it was stable between pH 4 and pH 11. 25341402 0 43 from site 135:138 arg1 secretion 147:155 its secretion 143:155 its secretion 143:155 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 0 43 from site 135:138 arg1 activity 161:168 activity 161:168 activity 161:168 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 1 44 contain have 195:198 arg2 potential 218:226 great application potential 200:226 great application potential 200:226 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 1 44 contain have 195:198 arg1 lyases 188:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 9 45 contain has 1230:1232 arg1 PelA 1225:1228 PelA 1225:1228 PelA 1225:1228 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 9 45 contain has 1230:1232 arg2 N-glycan 1243:1250 only one N-glycan 1234:1250 only one N-glycan attached to the residue N95 1234:1278 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 11 46 from N198 1458:1461 arg1 substitution 1430:1441 Amino acid substitution 1419:1441 Amino acid substitution at the residue N198 1419:1461 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 3 47 from volvacea 445:452 arg1 PelA 405:408 a novel alkaline pectate lyase PelA 374:408 a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 374:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 3 47 from volvacea 445:452 arg1 characteristics 355:369 the biochemical characteristics 339:369 the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 339:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 48 contain containing 513:522 arg1 polypeptide 501:511 a 321-amino-acid polypeptide 484:511 a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain 484:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 4 48 contain containing 513:522 arg2 peptide 553:559 a putative 18-residue signal peptide 524:559 a putative 18-residue signal peptide 524:559 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 4 48 contain containing 513:522 arg2 domain 600:605 a pectate lyase family 1 catalytic domain 565:605 a pectate lyase family 1 catalytic domain 565:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 4 48 contain containing 513:522 arg1 pelA 471:474 The full-length pelA 455:474 The full-length pelA 455:474 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 1 49 theme textiles 250:257 arg1 bioscouring 235:245 the bioscouring 231:245 the bioscouring of textiles 231:257 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 4 50 theme catalytic 590:598 arg1 domain 600:605 a pectate lyase family 1 catalytic domain 565:605 a pectate lyase family 1 catalytic domain 565:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 10 51 theme PelA 1413:1416 arg1 structure 1400:1408 the secondary structure 1386:1408 the secondary structure of PelA 1386:1416 This N-glycan is crucial to its efficient secretion and activity possibly due to its role in maintaining the secondary structure of PelA. 25341402 9 52 attach attached 1252:1259 arg1 N95 1276:1278 the residue N95 1264:1278 the residue N95 1264:1278 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 9 52 attach attached 1252:1259 arg2 N-glycan 1243:1250 only one N-glycan 1234:1250 only one N-glycan attached to the residue N95 1234:1278 Site-directed mutagenesis revealed PelA has only one N-glycan attached to the residue N95. 25341402 5 53 theme potential 656:664 arg1 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 53 theme potential 656:664 arg1 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 11 54 theme Amino 1419:1423 arg1 substitution 1430:1441 Amino acid substitution 1419:1441 Amino acid substitution at the residue N198 1419:1461 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 1 55 theme pectate 180:186 arg1 lyases 188:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases 171:193 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 5 56 contain contains 611:618 arg1 N198 721:724 N198 721:724 N198 721:724 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg1 It 608:609 It 608:609 It 608:609 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg2 site 682:685 one conserved and one non-conserved potential N-glycosylation site 620:685 one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) 620:695 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg2 N-X-S/T 688:694 N-X-S/T 688:694 N-X-S/T 688:694 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg1 residues 704:711 the residues N95 and N198 700:724 the residues N95 and N198 700:724 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg1 N95 713:715 N95 713:715 N95 713:715 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 56 contain contains 611:618 arg2 It 608:609 It 608:609 It 608:609 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 0 57 theme N-glycosylation 119:133 arg1 site 135:138 the highly conserved N-glycosylation site 98:138 the highly conserved N-glycosylation site in its secretion and activity 98:168 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 8 58 theme 0.1 1106:1108 arg1 mM 1110:1111 mM 1110:1111 mM 1110:1111 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 8 59 theme m 1135:1135 arg1 values 1137:1142 the K m values 1129:1142 the K m values 1129:1142 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 1 60 theme application 206:216 arg1 potential 218:226 great application potential 200:226 great application potential 200:226 Alkaline pectate lyases have great application potential in the bioscouring of textiles. 25341402 8 61 theme Ca 1113:1114 arg1 addition 1094:1101 the addition 1090:1101 the addition of 0.1 mM Ca(2+) 1090:1118 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 5 62 dep residues 704:711 arg1 residues 704:711 the residues N95 and N198 700:724 the residues N95 and N198 700:724 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 62 dep residues 704:711 arg1 N198 721:724 N198 721:724 N198 721:724 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 5 62 dep residues 704:711 arg1 N95 713:715 N95 713:715 N95 713:715 It contains one conserved and one non-conserved potential N-glycosylation site (N-X-S/T) at the residues N95 and N198, respectively. 25341402 3 63 from characteristics 355:369 arg1 volvacea 445:452 the basidiomycete Volvariella volvacea 415:452 the basidiomycete Volvariella volvacea 415:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 7 64 theme thermal 965:971 arg1 stability 973:981 thermal stability 965:981 thermal stability 965:981 Additional Ca(2+) was not required to measure PelA activity in vitro, but it could significantly enhance its activity and thermal stability. 25341402 2 65 attach isolated 269:276 arg1 bacteria 297:304 bacteria 297:304 bacteria 297:304 They are isolated predominantly from bacteria and a few fungi. 25341402 2 65 attach isolated 269:276 arg2 They 260:263 They 260:263 They 260:263 They are isolated predominantly from bacteria and a few fungi. 25341402 2 65 attach isolated 269:276 arg1 fungi 316:320 a few fungi 310:320 a few fungi 310:320 They are isolated predominantly from bacteria and a few fungi. 25341402 0 66 theme Volvariella 67:77 arg1 volvacea 79:86 Volvariella volvacea 67:86 Volvariella volvacea 67:86 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 11 67 theme catalytic 1741:1749 arg1 site 1751:1754 the catalytic site 1737:1754 the catalytic site 1737:1754 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 8 68 theme mg 1179:1180 arg1 ml 1182:1183 0.681 to 0.514 mg ml(-1) 1164:1187 0.681 to 0.514 mg ml(-1) 1164:1187 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 4 69 theme 18-residue 535:544 arg1 peptide 553:559 a putative 18-residue signal peptide 524:559 a putative 18-residue signal peptide 524:559 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 8 70 theme V 988:988 arg1 values 994:999 The V max values 984:999 The V max values using polygalacturonic acid as substrate 984:1040 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 8 71 dep mg 1080:1081 arg1 IU 1077:1078 IU 1077:1078 IU 1077:1078 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 8 72 theme polygalacturonic 1007:1022 arg1 acid 1024:1027 polygalacturonic acid 1007:1027 polygalacturonic acid 1007:1027 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 3 73 theme alkaline 382:389 arg1 PelA 405:408 a novel alkaline pectate lyase PelA 374:408 a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 374:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 4 74 theme lyase 575:579 arg1 domain 600:605 a pectate lyase family 1 catalytic domain 565:605 a pectate lyase family 1 catalytic domain 565:605 The full-length pelA encodes a 321-amino-acid polypeptide containing a putative 18-residue signal peptide and a pectate lyase family 1 catalytic domain. 25341402 8 75 dep 89.96 1071:1075 arg1 to 1068:1069 to 1068:1069 to 1068:1069 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 3 76 theme basidiomycete 419:431 arg1 volvacea 445:452 the basidiomycete Volvariella volvacea 415:452 the basidiomycete Volvariella volvacea 415:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 0 77 theme pectate 43:49 arg1 PelA 57:60 an alkaline pectate lyase PelA 31:60 an alkaline pectate lyase PelA 31:60 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 0 78 theme PelA 57:60 arg1 characteristics 12:26 Biochemical characteristics 0:26 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 3 79 theme lyase 399:403 arg1 PelA 405:408 a novel alkaline pectate lyase PelA 374:408 a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea 374:452 Here, we report the biochemical characteristics of a novel alkaline pectate lyase PelA from the basidiomycete Volvariella volvacea. 25341402 10 80 theme efficient 1313:1321 arg1 secretion 1323:1331 its efficient secretion 1309:1331 its efficient secretion 1309:1331 This N-glycan is crucial to its efficient secretion and activity possibly due to its role in maintaining the secondary structure of PelA. 25341402 11 81 theme acid 1425:1428 arg1 substitution 1430:1441 Amino acid substitution 1419:1441 Amino acid substitution at the residue N198 1419:1461 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 8 82 dep 0.514 1173:1177 arg1 to 1170:1171 to 1170:1171 to 1170:1171 The V max values using polygalacturonic acid as substrate were increased from 50.71 to 89.96 IU mg(-1) by the addition of 0.1 mM Ca(2+), whereas the K m values were decreased from 0.681 to 0.514 mg ml(-1). 25341402 11 83 dep slight 1514:1519 arg1 % 1547:1547 27.37 % 1541:1547 27.37 % 1541:1547 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 83 dep slight 1514:1519 arg1 to 1530:1531 to 1530:1531 to 1530:1531 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 83 dep slight 1514:1519 arg1 % 1527:1527 5.16 % 1522:1527 5.16 % 1522:1527 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 83 dep slight 1514:1519 arg1 modest 1533:1538 modest 1533:1538 modest 1533:1538 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 0 84 dep characteristics 12:26 arg1 roles 89:93 roles 89:93 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 0:169 Biochemical characteristics of an alkaline pectate lyase PelA from Volvariella volvacea: roles of the highly conserved N-glycosylation site in its secretion and activity. 25341402 10 85 theme secondary 1390:1398 arg1 structure 1400:1408 the secondary structure 1386:1408 the secondary structure of PelA 1386:1416 This N-glycan is crucial to its efficient secretion and activity possibly due to its role in maintaining the secondary structure of PelA. 25341402 11 86 theme PelA 1480:1483 arg1 secretion 1485:1493 PelA secretion 1480:1493 PelA secretion 1480:1493 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 87 theme residue 1450:1456 arg1 N198 1458:1461 the residue N198 1446:1461 the residue N198 1446:1461 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 88 theme due 1672:1674 arg1 activity 1663:1670 activity 1663:1670 activity due to it being highly conserved 1663:1703 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 89 theme amino 1623:1627 arg1 acid 1629:1632 the amino acid itself 1619:1639 the amino acid itself 1619:1639 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 25341402 11 89 theme amino 1623:1627 arg1 important 1649:1657 important 1649:1657 important 1649:1657 Amino acid substitution at the residue N198 had no effect on PelA secretion, but resulted in a slight (5.16 %) to modest (27.37 %) decrease in specific activity and less thermal stability, indicating the amino acid itself is also important for activity due to it being highly conserved and because of its proximity to the catalytic site. 24467287 7 0 theme cell 1396:1399 arg1 lines 1401:1405 both rodent and mosquito cell lines 1371:1405 both rodent and mosquito cell lines 1371:1405 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 4 1 theme prototypic 613:622 arg1 virus 651:655 the prototypic alphavirus, Semliki Forest virus 609:655 virus 651:655 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 7 2 theme protruding 1241:1250 arg1 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 6 3 link mosquito-derived 1129:1144 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 5 4 gly glycoprotein 894:905 arg1 glycoprotein 894:905 the protruding E2 attachment glycoprotein 865:905 the protruding E2 attachment glycoprotein 865:905 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 4 5 theme rodent 691:696 arg1 lines 703:707 both arthropod and rodent cell lines 672:707 both arthropod and rodent cell lines 672:707 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 2 6 link N-linked 410:417 arg1 glycans 419:425 N-linked glycans 410:425 N-linked glycans 410:425 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 2 7 theme virus 318:322 arg1 E1 324:325 Semliki Forest virus E1 303:325 Semliki Forest virus E1 303:325 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 1 8 theme viral 213:217 arg1 glycoproteins 228:240 viral envelope glycoproteins 213:240 viral envelope glycoproteins 213:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 11 9 theme Semliki 1991:1997 arg1 virus 2006:2010 Semliki Forest virus 1991:2010 Semliki Forest virus 1991:2010 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 1 10 theme glycoproteins 228:240 arg1 processing 199:208 differential post-translational processing 167:208 differential post-translational processing of viral envelope glycoproteins 167:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 6 11 theme processed 1067:1075 arg1 glycans 1077:1083 highly processed glycans 1060:1083 highly processed glycans dependent on the host cell 1060:1110 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 4 12 theme arthropod 677:685 arg1 lines 703:707 both arthropod and rodent cell lines 672:707 both arthropod and rodent cell lines 672:707 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 10 13 theme surface 1801:1807 arg1 model 1761:1765 a detailed model 1750:1765 a detailed model of the glycosylated mature virion surface 1750:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 5 14 theme heterogeneous 915:927 arg1 glycosylation 929:941 heterogeneous glycosylation 915:941 heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing 915:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 10 15 theme mature 1787:1792 arg1 surface 1801:1807 the glycosylated mature virion surface 1770:1807 the glycosylated mature virion surface 1770:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 11 16 theme virus 2006:2010 arg1 transmission 1975:1986 interspecies transmission 1962:1986 interspecies transmission of Semliki Forest virus 1962:2010 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 5 17 theme extensive 1002:1010 arg1 processing 1012:1021 both limited and extensive processing 985:1021 both limited and extensive processing 985:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 4 18 theme collision-induced 754:770 arg1 dissociation 772:783 collision-induced dissociation 754:783 collision-induced dissociation 754:783 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 2 19 theme Semliki 303:309 arg1 virus 318:322 Semliki Forest virus 303:322 Semliki Forest virus E1 303:325 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 1 20 theme progeny 143:149 arg1 alphaviruses 151:162 parent and progeny alphaviruses 132:162 parent and progeny alphaviruses 132:162 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 11 21 theme processing 1911:1920 arg1 enzymes 1922:1928 cellular processing enzymes 1902:1928 cellular processing enzymes 1902:1928 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 7 22 theme mosquito 1387:1394 arg1 lines 1401:1405 both rodent and mosquito cell lines 1371:1405 both rodent and mosquito cell lines 1371:1405 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 7 23 theme conserved 1299:1307 arg1 structures 1343:1352 conserved under-processed oligomannose-type structures 1299:1352 conserved under-processed oligomannose-type structures 1299:1352 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 1 24 theme differential 167:178 arg1 processing 199:208 differential post-translational processing 167:208 differential post-translational processing of viral envelope glycoproteins 167:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 4 25 theme ion-mobility 719:730 arg1 spectrometry 737:748 ion-mobility mass spectrometry 719:748 ion-mobility mass spectrometry 719:748 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 7 26 theme rodent 1376:1381 arg1 lines 1401:1405 both rodent and mosquito cell lines 1371:1405 both rodent and mosquito cell lines 1371:1405 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 4 27 from composition 577:587 arg1 virus 651:655 the prototypic alphavirus, Semliki Forest virus 609:655 virus 651:655 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 5 28 theme limited 990:996 arg1 processing 1012:1021 both limited and extensive processing 985:1021 both limited and extensive processing 985:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 5 29 contain contains 948:955 arg2 glycans 966:972 N-linked glycans 957:972 N-linked glycans exhibiting both limited and extensive processing 957:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 5 29 contain contains 948:955 arg1 glycosylation 929:941 heterogeneous glycosylation 915:941 heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing 915:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 11 30 theme virion 1940:1945 arg1 glycome 1947:1953 the virion glycome 1936:1953 the virion glycome 1936:1953 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 6 31 theme dependent 1085:1093 arg1 glycans 1077:1083 highly processed glycans 1060:1083 highly processed glycans dependent on the host cell 1060:1110 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 8 32 theme viral 1514:1518 arg1 structure 1528:1536 local viral protein structure 1508:1536 local viral protein structure 1508:1536 It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. 24467287 5 33 link N-linked 957:964 arg1 glycans 966:972 N-linked glycans 957:972 N-linked glycans exhibiting both limited and extensive processing 957:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 6 34 theme paucimannose-type 1177:1193 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 10 35 gly glycosylated 1774:1785 arg1 surface 1801:1807 the glycosylated mature virion surface 1770:1807 the glycosylated mature virion surface 1770:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 0 36 theme Structural 0:9 arg1 plasticity 11:20 Structural plasticity 0:20 Structural plasticity of the Semliki Forest virus 0:48 Structural plasticity of the Semliki Forest virus glycome upon interspecies transmission. 24467287 1 37 theme viral 104:108 arg1 transmission 110:121 Cross-species viral transmission 90:121 Cross-species viral transmission 90:121 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 6 38 theme complex-type 1160:1171 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 3 39 theme host 508:511 arg1 cells 513:517 different host cells 498:517 different host cells 498:517 However the composition of these glycans, including that arising from different host cells, has not been determined. 24467287 5 40 theme E2 880:881 arg1 glycoprotein 894:905 the protruding E2 attachment glycoprotein 865:905 the protruding E2 attachment glycoprotein 865:905 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 6 41 theme E1 1146:1147 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 4 42 theme glycans 596:602 arg1 composition 577:587 the chemical composition 564:587 the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, 564:708 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 7 43 theme oligomannose-type 1325:1341 arg1 structures 1343:1352 conserved under-processed oligomannose-type structures 1299:1352 conserved under-processed oligomannose-type structures 1299:1352 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 0 44 theme Semliki 29:35 arg1 virus 44:48 the Semliki Forest virus 25:48 the Semliki Forest virus 25:48 Structural plasticity of the Semliki Forest virus glycome upon interspecies transmission. 24467287 5 45 theme E1 838:839 arg1 glycoprotein 848:859 the membrane-proximal E1 fusion glycoprotein 816:859 the membrane-proximal E1 fusion glycoprotein 816:859 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 10 46 theme cryo-electron 1684:1696 arg1 microscopy 1698:1707 previous cryo-electron microscopy 1675:1707 previous cryo-electron microscopy 1675:1707 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 4 47 from virus 651:655 arg1 composition 577:587 the chemical composition 564:587 the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, 564:708 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 4 47 from virus 651:655 arg1 glycans 596:602 the glycans 592:602 the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, 592:708 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 0 48 theme virus 44:48 arg1 plasticity 11:20 Structural plasticity 0:20 Structural plasticity of the Semliki Forest virus 0:48 Structural plasticity of the Semliki Forest virus glycome upon interspecies transmission. 24467287 4 49 theme chemical 568:575 arg1 composition 577:587 the chemical composition 564:587 the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, 564:708 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 11 50 theme cellular 1902:1909 arg1 enzymes 1922:1928 cellular processing enzymes 1902:1928 cellular processing enzymes 1902:1928 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 5 51 gly glycoprotein 848:859 arg1 glycoprotein 848:859 the membrane-proximal E1 fusion glycoprotein 816:859 the membrane-proximal E1 fusion glycoprotein 816:859 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 2 52 theme N-linked 410:417 arg1 glycans 419:425 N-linked glycans 410:425 N-linked glycans 410:425 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 9 53 theme host 1603:1606 arg1 cell 1608:1611 the host cell 1599:1611 the host cell 1599:1611 This contrasts E1, which presents glycans characteristic of the host cell and is accessible to enzymes. 24467287 5 54 theme fusion 841:846 arg1 glycoprotein 848:859 the membrane-proximal E1 fusion glycoprotein 816:859 the membrane-proximal E1 fusion glycoprotein 816:859 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 2 55 gly glycoproteins 334:346 arg1 glycoproteins 334:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 2 56 theme processing 396:405 arg1 degrees 385:391 differing degrees 375:391 differing degrees of processing of N-linked glycans 375:425 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 10 57 theme detailed 1752:1759 arg1 model 1761:1765 a detailed model 1750:1765 a detailed model of the glycosylated mature virion surface 1750:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 1 58 gly glycoproteins 228:240 arg1 glycoproteins 228:240 viral envelope glycoproteins 213:240 viral envelope glycoproteins 213:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 2 59 theme E2 331:332 arg1 glycoproteins 334:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 9 60 theme characteristic 1581:1594 arg1 glycans 1573:1579 glycans 1573:1579 glycans characteristic of the host cell 1573:1611 This contrasts E1, which presents glycans characteristic of the host cell and is accessible to enzymes. 24467287 4 61 theme alphavirus 624:633 arg1 virus 651:655 the prototypic alphavirus, Semliki Forest virus 609:655 virus 651:655 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 2 62 theme E1 324:325 arg1 glycoproteins 334:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 the Semliki Forest virus E1 and E2 glycoproteins 299:346 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 9 63 theme cell 1608:1611 arg1 characteristic 1581:1594 characteristic 1581:1594 characteristic 1581:1594 This contrasts E1, which presents glycans characteristic of the host cell and is accessible to enzymes. 24467287 7 64 theme attachment 1255:1264 arg1 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 10 65 theme glycosylated 1774:1785 arg1 surface 1801:1807 the glycosylated mature virion surface 1770:1807 the glycosylated mature virion surface 1770:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 7 66 contain contained 1289:1297 arg1 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 7 66 contain contained 1289:1297 arg2 structures 1343:1352 conserved under-processed oligomannose-type structures 1299:1352 conserved under-processed oligomannose-type structures 1299:1352 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 7 66 contain contained 1289:1297 arg1 contrast 1227:1234 contrast 1227:1234 contrast 1227:1234 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 7 66 contain contained 1289:1297 arg2 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 1 67 theme envelope 219:226 arg1 glycoproteins 228:240 viral envelope glycoproteins 213:240 viral envelope glycoproteins 213:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 4 68 theme cell 698:701 arg1 lines 703:707 both arthropod and rodent cell lines 672:707 both arthropod and rodent cell lines 672:707 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 11 69 theme Forest 1999:2004 arg1 virus 2006:2010 Semliki Forest virus 1991:2010 Semliki Forest virus 1991:2010 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 5 70 theme attachment 883:892 arg1 glycoprotein 894:905 the protruding E2 attachment glycoprotein 865:905 the protruding E2 attachment glycoprotein 865:905 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 5 71 theme N-linked 957:964 arg1 glycans 966:972 N-linked glycans 957:972 N-linked glycans exhibiting both limited and extensive processing 957:1021 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 10 72 theme virion 1794:1799 arg1 surface 1801:1807 the glycosylated mature virion surface 1770:1807 the glycosylated mature virion surface 1770:1807 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 1 73 theme parent 132:137 arg1 alphaviruses 151:162 parent and progeny alphaviruses 132:162 parent and progeny alphaviruses 132:162 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 4 74 theme mass 732:735 arg1 spectrometry 737:748 ion-mobility mass spectrometry 719:748 ion-mobility mass spectrometry 719:748 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 4 75 dep virus 651:655 arg1 Semliki 636:642 Semliki 636:642 Semliki 636:642 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 2 76 theme Forest 311:316 arg1 virus 318:322 Semliki Forest virus 303:322 Semliki Forest virus E1 303:325 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 0 77 theme Forest 37:42 arg1 virus 44:48 the Semliki Forest virus 25:48 the Semliki Forest virus 25:48 Structural plasticity of the Semliki Forest virus glycome upon interspecies transmission. 24467287 8 78 theme E2 1447:1448 arg1 processing 1433:1442 glycan processing 1426:1442 glycan processing of E2 1426:1448 It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. 24467287 0 79 theme interspecies 63:74 arg1 transmission 76:87 interspecies transmission 63:87 interspecies transmission 63:87 Structural plasticity of the Semliki Forest virus glycome upon interspecies transmission. 24467287 1 80 theme post-translational 180:197 arg1 processing 199:208 differential post-translational processing 167:208 differential post-translational processing of viral envelope glycoproteins 167:240 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 6 81 theme host 1102:1105 arg1 cell 1107:1110 the host cell 1098:1110 the host cell 1098:1110 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 11 82 theme protein 1880:1886 arg1 structure 1888:1896 virally encoded protein structure 1864:1896 virally encoded protein structure 1864:1896 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 11 83 theme interspecies 1962:1973 arg1 transmission 1975:1986 interspecies transmission 1962:1986 interspecies transmission of Semliki Forest virus 1962:2010 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 8 84 theme local 1508:1512 arg1 structure 1528:1536 local viral protein structure 1508:1536 local viral protein structure 1508:1536 It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. 24467287 7 85 theme under-processed 1309:1323 arg1 structures 1343:1352 conserved under-processed oligomannose-type structures 1299:1352 conserved under-processed oligomannose-type structures 1299:1352 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 1 86 theme Cross-species 90:102 arg1 transmission 110:121 Cross-species viral transmission 90:121 Cross-species viral transmission 90:121 Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. 24467287 8 87 theme protein 1520:1526 arg1 structure 1528:1536 local viral protein structure 1508:1536 local viral protein structure 1508:1536 It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. 24467287 2 88 theme glycans 419:425 arg1 processing 396:405 processing 396:405 processing of N-linked glycans 396:425 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 5 89 theme protruding 869:878 arg1 glycoprotein 894:905 the protruding E2 attachment glycoprotein 865:905 the protruding E2 attachment glycoprotein 865:905 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 4 90 dep spectrometry 737:748 arg1 analysis 785:792 analysis 785:792 analysis 785:792 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 6 91 theme exhibiting 1149:1158 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 5 92 theme membrane-proximal 820:836 arg1 glycoprotein 848:859 the membrane-proximal E1 fusion glycoprotein 816:859 the membrane-proximal E1 fusion glycoprotein 816:859 We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. 24467287 8 93 theme glycan 1426:1431 arg1 processing 1433:1442 glycan processing 1426:1442 glycan processing of E2 1426:1448 It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. 24467287 11 94 theme encoded 1872:1878 arg1 structure 1888:1896 virally encoded protein structure 1864:1896 virally encoded protein structure 1864:1896 Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus. 24467287 6 95 theme mosquito-derived 1129:1144 arg1 glycosylation 1195:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation 1129:1207 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 3 96 theme glycans 461:467 arg1 composition 440:450 the composition 436:450 the composition of these glycans, including that arising from different host cells, 436:518 However the composition of these glycans, including that arising from different host cells, has not been determined. 24467287 7 97 gly glycoprotein 1266:1277 arg1 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 3 98 theme different 498:506 arg1 cells 513:517 different host cells 498:517 different host cells 498:517 However the composition of these glycans, including that arising from different host cells, has not been determined. 24467287 7 99 theme E2 1252:1253 arg1 glycoprotein 1266:1277 the protruding E2 attachment glycoprotein 1237:1277 the protruding E2 attachment glycoprotein 1237:1277 In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. 24467287 10 100 theme previous 1675:1682 arg1 microscopy 1698:1707 previous cryo-electron microscopy 1675:1707 previous cryo-electron microscopy 1675:1707 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 2 101 theme differing 375:383 arg1 degrees 385:391 differing degrees 375:391 differing degrees of processing of N-linked glycans 375:425 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 4 102 theme Forest 644:649 arg1 virus 651:655 the prototypic alphavirus, Semliki Forest virus 609:655 virus 651:655 Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. 24467287 2 103 theme Alphavirus 243:252 arg1 biogenesis 254:263 Alphavirus biogenesis 243:263 Alphavirus biogenesis 243:263 Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. 24467287 10 104 theme crystallographic 1713:1728 arg1 analyses 1730:1737 crystallographic analyses 1713:1737 crystallographic analyses 1713:1737 We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. 24467287 6 105 contain contained 1036:1044 arg1 E1 1033:1034 E1 1033:1034 E1 1033:1034 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 24467287 6 105 contain contained 1036:1044 arg2 glycans 1077:1083 highly processed glycans 1060:1083 highly processed glycans dependent on the host cell 1060:1110 However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. 25895110 7 0 from decrease 1133:1140 arg1 proportion 1149:1158 the proportion 1145:1158 the proportion of sialylated structures in CD 1145:1189 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 5 1 with patients 632:639 arg1 CD 660:661 CD 660:661 CD (n = 287) 660:671 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 1 with patients 632:639 arg1 controls 678:685 controls 678:685 controls (n = 320) 678:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 1 with patients 632:639 arg1 n 650:650 n = 507 650:656 n = 507 650:656 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 1 with patients 632:639 arg1 n 688:688 n = 320 688:694 n = 320 688:694 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 1 with patients 632:639 arg1 287 668:670 n = 287 664:670 n = 287 664:670 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 1 with patients 632:639 arg1 UC 646:647 UC 646:647 UC (n = 507) 646:657 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 9 2 dep increased 1655:1663 arg1 compared 1706:1713 compared 1706:1713 compared with no colectomy (FDR-adjusted, P = 0.05) 1706:1756 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 7 3 theme = 1018:1018 arg1 odds 1002:1005 odds 1002:1005 odds ratio [OR] = 0.71; 95% confidence interval [CI 1002:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 4 dep galactosylation 962:976 arg1 digalactosylation 979:995 digalactosylation 979:995 digalactosylation 979:995 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 4 dep galactosylation 962:976 arg1 = 1105:1105 = 1105:1105 = 1105:1105 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 5 5 theme glycome 609:615 arg1 composition 617:627 METHODS IgG glycome composition 597:627 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) 597:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 7 6 theme P 1216:1216 arg1 × 1224:1224 = 0.46, CI, 0.3-0.6, P = 8.4 × 10 1195:1227 × 1224:1224 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 7 theme = 1195:1195 arg1 × 1224:1224 = 0.46, CI, 0.3-0.6, P = 8.4 × 10 1195:1227 × 1224:1224 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 8 dep = 1018:1018 arg1 [CI 1050:1052 0.71; 95% confidence interval [CI 1020:1052 0.71; 95% confidence interval [CI 1020:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 9 9 with patients 1668:1675 arg1 UC 1682:1683 UC 1682:1683 UC 1682:1683 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 2 10 theme immunoglobulin 312:325 arg1 IgG 330:332 IgG 330:332 IgG 330:332 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 2 10 theme immunoglobulin 312:325 arg1 G 327:327 immunoglobulin G 312:327 the immunoglobulin G (IgG) Fc fragment 308:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 8 11 theme characteristic 1414:1427 arg1 curves 1429:1434 receiver-operator characteristic curves 1396:1434 receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1396:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 9 12 dep = 1750:1750 arg1 FDR-adjusted 1734:1745 FDR-adjusted 1734:1745 FDR-adjusted 1734:1745 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 6 13 from differences 788:798 arg1 composition 815:825 IgG glycome composition 803:825 IgG glycome composition 803:825 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 10 14 theme observed 1775:1782 arg1 differences 1784:1794 The observed differences 1771:1794 The observed differences 1771:1794 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 10 15 from potential 1842:1850 arg1 IBD 1862:1864 IBD 1862:1864 IBD 1862:1864 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 8 16 theme = 1551:1551 arg1 P 1549:1549 P 1549:1549 P = 0.026 1549:1557 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 2 17 attach attached 296:303 arg1 fragment 338:345 the immunoglobulin G (IgG) Fc fragment 308:345 the immunoglobulin G (IgG) Fc fragment 308:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 2 17 attach attached 296:303 arg2 N-glycans 286:294 N-glycans 286:294 N-glycans attached to the immunoglobulin G (IgG) Fc fragment 286:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 12 18 theme IgG 1989:1991 arg1 profiles 2000:2007 IgG glycan profiles 1989:2007 IgG glycan profiles 1989:2007 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 7 19 theme sialylated 1163:1172 arg1 structures 1174:1183 sialylated structures 1163:1183 sialylated structures 1163:1183 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 5 20 theme = 652:652 arg1 n 650:650 n = 507 650:656 n = 507 650:656 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 20 theme = 652:652 arg1 UC 646:647 UC 646:647 UC (n = 507) 646:657 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 1 21 theme underexplored 132:144 arg1 area 155:158 an underexplored research area 129:158 an underexplored research area in inflammatory bowel disease (IBD) 129:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 21 theme underexplored 132:144 arg1 Glycobiology 113:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 7 22 theme % 1028:1028 arg1 [CI 1050:1052 0.71; 95% confidence interval [CI 1020:1052 0.71; 95% confidence interval [CI 1020:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 23 theme decreased 948:956 arg1 galactosylation 962:976 significantly decreased IgG galactosylation 934:976 significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) 934:1115 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 8 24 theme model 1479:1483 arg1 performance 1457:1467 better performance 1450:1467 better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1450:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 10 25 theme inflammatory 1829:1840 arg1 potential 1842:1850 significantly increased inflammatory potential 1805:1850 significantly increased inflammatory potential of IgG in IBD 1805:1864 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 1 26 theme inflammatory 163:174 arg1 IBD 191:193 IBD 191:193 IBD 191:193 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 26 theme inflammatory 163:174 arg1 disease 182:188 inflammatory bowel disease 163:188 inflammatory bowel disease (IBD) 163:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 8 27 theme Logistic 1231:1238 arg1 models 1251:1256 Logistic regression models 1231:1256 Logistic regression models incorporating measured IgG glycan traits 1231:1297 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 9 28 theme GlcNAc 1611:1616 arg1 absence 1590:1596 absence 1590:1596 absence of bisecting GlcNAc in monogalactosylated structures 1590:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 0 29 theme G 91:91 arg1 glycome 93:99 the immunoglobulin G glycome 72:99 the immunoglobulin G glycome 72:99 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 8 30 dep = 1355:1355 arg1 UC 1349:1350 UC 1349:1350 UC 1349:1350 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 31 theme curve 1497:1501 arg1 [AUC 1503:1506 curve [AUC 1497:1506 curve [AUC 1497:1506 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 4 32 theme IgG 525:527 arg1 changes 536:542 IgG glycan changes 525:542 IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD) 525:594 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 9 33 theme monogalactosylated 1621:1638 arg1 structures 1640:1649 monogalactosylated structures 1621:1649 monogalactosylated structures 1621:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 8 34 theme measured 1272:1279 arg1 traits 1292:1297 measured IgG glycan traits 1272:1297 measured IgG glycan traits 1272:1297 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 12 35 contain have 2009:2012 arg1 profiles 2000:2007 IgG glycan profiles 1989:2007 IgG glycan profiles 1989:2007 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 12 35 contain have 2009:2012 arg2 biomarkers 2045:2054 IBD biomarkers 2041:2054 IBD biomarkers 2041:2054 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 12 35 contain have 2009:2012 arg2 potential 2028:2036 translational potential 2014:2036 translational potential 2014:2036 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 0 36 theme proinflammatory 43:57 arg1 potential 59:67 proinflammatory potential 43:67 proinflammatory potential of the immunoglobulin G glycome 43:99 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 7 37 from proportion 1149:1158 arg1 CD 1188:1189 CD 1188:1189 CD 1188:1189 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 9 38 from GlcNAc 1611:1616 arg1 structures 1640:1649 monogalactosylated structures 1621:1649 monogalactosylated structures 1621:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 8 39 theme glycan 1285:1290 arg1 traits 1292:1297 measured IgG glycan traits 1272:1297 measured IgG glycan traits 1272:1297 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 0 40 theme Inflammatory 0:11 arg1 disease 19:25 Inflammatory bowel disease 0:25 Inflammatory bowel disease 0:25 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 5 41 from composition 617:627 arg1 patients 632:639 patients 632:639 patients with UC (n = 507), CD (n = 287), and controls (n = 320) 632:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 1 42 theme BACKGROUND 102:111 arg1 area 155:158 an underexplored research area 129:158 an underexplored research area in inflammatory bowel disease (IBD) 129:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 42 theme BACKGROUND 102:111 arg1 Glycobiology 113:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 5 43 theme performance 719:729 arg1 chromatography 738:751 ultra performance liquid chromatography 713:751 ultra performance liquid chromatography 713:751 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 3 44 theme genome-wide 413:423 arg1 studies 437:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies reveal pleiotropy between IBD and IgG glycosylation. 25895110 11 45 from Changes 1867:1873 arg1 glycosylation 1882:1894 IgG glycosylation 1878:1894 IgG glycosylation 1878:1894 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 7 46 theme interval 1041:1048 arg1 [CI 1050:1052 0.71; 95% confidence interval [CI 1020:1052 0.71; 95% confidence interval [CI 1020:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 8 47 dep = 1551:1551 arg1 0.026 1553:1557 0.026 1553:1557 0.026 1553:1557 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 6 48 theme significant 776:786 arg1 differences 788:798 Statistically significant differences 762:798 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, 754:881 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 2 49 theme Fc 335:336 arg1 fragment 338:345 the immunoglobulin G (IgG) Fc fragment 308:345 the immunoglobulin G (IgG) Fc fragment 308:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 4 50 from changes 536:542 arg1 disease 583:589 Crohn's disease 575:589 Crohn's disease (CD) 575:594 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 4 50 from changes 536:542 arg1 UC 567:568 UC 567:568 UC 567:568 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 4 50 from changes 536:542 arg1 colitis 558:564 ulcerative colitis 547:564 ulcerative colitis (UC) 547:569 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 4 50 from changes 536:542 arg1 CD 592:593 CD 592:593 CD 592:593 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 10 51 dep CONCLUSIONS 1759:1769 arg1 indicate 1796:1803 indicate 1796:1803 indicate significantly increased inflammatory potential of IgG in IBD 1796:1864 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 6 52 with patients 835:842 arg1 CD 855:856 CD 855:856 CD 855:856 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 6 52 with patients 835:842 arg1 UC 849:850 UC 849:850 UC 849:850 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 11 53 theme antibody 1958:1965 arg1 efficacy 1979:1986 monoclonal antibody therapeutic efficacy 1947:1986 monoclonal antibody therapeutic efficacy 1947:1986 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 4 54 theme ulcerative 547:556 arg1 UC 567:568 UC 567:568 UC 567:568 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 4 54 theme ulcerative 547:556 arg1 colitis 558:564 ulcerative colitis 547:564 ulcerative colitis (UC) 547:569 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 10 55 theme increased 1819:1827 arg1 potential 1842:1850 significantly increased inflammatory potential 1805:1850 significantly increased inflammatory potential of IgG in IBD 1805:1864 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 8 56 dep model 1479:1483 arg1 area 1486:1489 area 1486:1489 area under curve [AUC 1486:1506 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 2 57 theme molecular 358:366 arg1 structure 368:376 molecular structure 358:376 molecular structure 358:376 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 1 58 theme etiological 230:240 arg1 mechanisms 242:251 many etiological mechanisms 225:251 many etiological mechanisms described in IBD 225:268 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 7 59 from galactosylation 962:976 arg1 proportion 1149:1158 the proportion 1145:1158 the proportion of sialylated structures in CD 1145:1189 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 8 60 theme UC 1526:1527 arg1 model 1529:1533 the UC model 1522:1533 the UC model (AUC = 0.72) (P = 0.026) 1522:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 60 theme UC 1526:1527 arg1 AUC 1536:1538 AUC = 0.72 1536:1545 AUC = 0.72 1536:1545 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 6 61 theme IgG 803:805 arg1 composition 815:825 IgG glycome composition 803:825 IgG glycome composition 803:825 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 7 62 theme = 1218:1218 arg1 × 1224:1224 = 0.46, CI, 0.3-0.6, P = 8.4 × 10 1195:1227 × 1224:1224 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 12 63 theme translational 2014:2026 arg1 potential 2028:2036 translational potential 2014:2036 translational potential 2014:2036 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 12 63 theme translational 2014:2026 arg1 biomarkers 2045:2054 IBD biomarkers 2041:2054 IBD biomarkers 2041:2054 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 5 64 theme IgG 605:607 arg1 composition 617:627 METHODS IgG glycome composition 597:627 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) 597:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 3 65 theme Recent 406:411 arg1 studies 437:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies reveal pleiotropy between IBD and IgG glycosylation. 25895110 6 66 theme glycome 807:813 arg1 composition 815:825 IgG glycome composition 803:825 IgG glycome composition 803:825 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 2 67 theme G 327:327 arg1 fragment 338:345 the immunoglobulin G (IgG) Fc fragment 308:345 the immunoglobulin G (IgG) Fc fragment 308:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 8 68 theme receiver-operator 1396:1412 arg1 curves 1429:1434 receiver-operator characteristic curves 1396:1434 receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1396:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 5 69 theme liquid 731:736 arg1 chromatography 738:751 ultra performance liquid chromatography 713:751 ultra performance liquid chromatography 713:751 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 11 70 theme IBD 1914:1916 arg1 pathogenesis 1918:1929 IBD pathogenesis 1914:1929 IBD pathogenesis 1914:1929 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 12 71 theme glycan 1993:1998 arg1 profiles 2000:2007 IgG glycan profiles 1989:2007 IgG glycan profiles 1989:2007 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 7 72 theme IgG 958:960 arg1 galactosylation 962:976 significantly decreased IgG galactosylation 934:976 significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) 934:1115 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 8 73 with able 1304:1307 arg1 curves 1429:1434 receiver-operator characteristic curves 1396:1434 receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1396:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 74 theme better 1450:1455 arg1 performance 1457:1467 better performance 1450:1467 better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1450:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 7 75 gly sialylated 1163:1172 arg1 structures 1174:1183 sialylated structures 1163:1183 sialylated structures 1163:1183 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 76 theme structures 1174:1183 arg1 proportion 1149:1158 the proportion 1145:1158 the proportion of sialylated structures in CD 1145:1189 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 9 77 theme presence 1578:1585 arg1 ratio 1565:1569 The ratio 1561:1569 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures 1561:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 8 78 theme = 1540:1540 arg1 model 1529:1533 the UC model 1522:1533 the UC model (AUC = 0.72) (P = 0.026) 1522:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 78 theme = 1540:1540 arg1 AUC 1536:1538 AUC = 0.72 1536:1545 AUC = 0.72 1536:1545 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 3 79 dep IBD 471:473 arg1 glycosylation 483:495 glycosylation 483:495 glycosylation 483:495 Recent genome-wide association studies reveal pleiotropy between IBD and IgG glycosylation. 25895110 1 80 theme research 146:153 arg1 area 155:158 an underexplored research area 129:158 an underexplored research area in inflammatory bowel disease (IBD) 129:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 80 theme research 146:153 arg1 Glycobiology 113:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology 102:124 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 2 81 from Alterations 271:281 arg1 N-glycans 286:294 N-glycans 286:294 N-glycans attached to the immunoglobulin G (IgG) Fc fragment 286:345 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 0 82 theme immunoglobulin 76:89 arg1 glycome 93:99 the immunoglobulin G glycome 72:99 the immunoglobulin G glycome 72:99 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 8 83 theme CD 1476:1477 arg1 model 1479:1483 the CD model 1472:1483 the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026) 1472:1558 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 84 theme regression 1240:1249 arg1 models 1251:1256 Logistic regression models 1231:1256 Logistic regression models incorporating measured IgG glycan traits 1231:1297 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 9 85 theme bisecting 1601:1609 arg1 GlcNAc 1611:1616 bisecting GlcNAc 1601:1616 bisecting GlcNAc in monogalactosylated structures 1601:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 11 86 theme IgG 1878:1880 arg1 glycosylation 1882:1894 IgG glycosylation 1878:1894 IgG glycosylation 1878:1894 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 0 87 theme glycome 93:99 arg1 potential 59:67 proinflammatory potential 43:67 proinflammatory potential of the immunoglobulin G glycome 43:99 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 8 88 dep 2.13 1357:1360 arg1 CD 1371:1372 CD 1371:1372 CD 1371:1372 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 88 dep 2.13 1357:1360 arg1 × 1362:1362 × 10 1362:1365 × 10 1362:1365 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 1 89 theme bowel 176:180 arg1 IBD 191:193 IBD 191:193 IBD 191:193 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 89 theme bowel 176:180 arg1 disease 182:188 inflammatory bowel disease 163:188 inflammatory bowel disease (IBD) 163:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 9 90 dep colectomy 1723:1731 arg1 = 1750:1750 = 1750:1750 = 1750:1750 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 4 91 theme glycan 529:534 arg1 changes 536:542 IgG glycan changes 525:542 IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD) 525:594 This study aims to explore IgG glycan changes in ulcerative colitis (UC) and Crohn's disease (CD). 25895110 5 92 theme = 666:666 arg1 CD 660:661 CD 660:661 CD (n = 287) 660:671 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 92 theme = 666:666 arg1 287 668:670 n = 287 664:670 n = 287 664:670 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 8 93 dep model 1529:1533 arg1 P 1549:1549 P 1549:1549 P = 0.026 1549:1557 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 8 94 theme IgG 1281:1283 arg1 traits 1292:1297 measured IgG glycan traits 1272:1297 measured IgG glycan traits 1272:1297 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 10 95 theme IgG 1855:1857 arg1 potential 1842:1850 significantly increased inflammatory potential 1805:1850 significantly increased inflammatory potential of IgG in IBD 1805:1864 CONCLUSIONS The observed differences indicate significantly increased inflammatory potential of IgG in IBD. 25895110 5 96 theme METHODS 597:603 arg1 composition 617:627 METHODS IgG glycome composition 597:627 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) 597:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 7 97 theme significant 1121:1131 arg1 decrease 1133:1140 significant decrease 1121:1140 significant decrease in the proportion of sialylated structures in CD 1121:1189 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 98 dep associated 918:927 arg1 × 1224:1224 = 0.46, CI, 0.3-0.6, P = 8.4 × 10 1195:1227 × 1224:1224 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 2 99 theme immunological 382:394 arg1 function 396:403 immunological function 382:403 immunological function 382:403 Alterations in N-glycans attached to the immunoglobulin G (IgG) Fc fragment can affect molecular structure and immunological function. 25895110 0 100 theme bowel 13:17 arg1 disease 19:25 Inflammatory bowel disease 0:25 Inflammatory bowel disease 0:25 Inflammatory bowel disease associates with proinflammatory potential of the immunoglobulin G glycome. 25895110 8 101 dep controls 1339:1346 arg1 = 1355:1355 = 1355:1355 = 1355:1355 Logistic regression models incorporating measured IgG glycan traits were able to distinguish UC and CD from controls (UC: P = 2.13 × 10 and CD: P = 2.20 × 10), with receiver-operator characteristic curves demonstrating better performance of the CD model (area under curve [AUC] = 0.77) over the UC model (AUC = 0.72) (P = 0.026). 25895110 1 102 from area 155:158 arg1 IBD 191:193 IBD 191:193 IBD 191:193 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 1 102 from area 155:158 arg1 disease 182:188 inflammatory bowel disease 163:188 inflammatory bowel disease (IBD) 163:194 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 9 103 from absence 1590:1596 arg1 structures 1640:1649 monogalactosylated structures 1621:1649 monogalactosylated structures 1621:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 5 104 theme ultra 713:717 arg1 chromatography 738:751 ultra performance liquid chromatography 713:751 ultra performance liquid chromatography 713:751 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 6 105 dep RESULTS 754:760 arg1 differences 788:798 Statistically significant differences 762:798 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, 754:881 RESULTS Statistically significant differences in IgG glycome composition between patients with UC or CD, compared with controls, were observed. 25895110 7 106 theme confidence 1030:1039 arg1 [CI 1050:1052 0.71; 95% confidence interval [CI 1020:1052 0.71; 95% confidence interval [CI 1020:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 3 107 theme association 425:435 arg1 studies 437:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies 406:443 Recent genome-wide association studies reveal pleiotropy between IBD and IgG glycosylation. 25895110 5 108 theme n 664:664 arg1 CD 660:661 CD 660:661 CD (n = 287) 660:671 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 108 theme n 664:664 arg1 287 668:670 n = 287 664:670 n = 287 664:670 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 7 109 dep = 1105:1105 arg1 = 1067:1067 = 1067:1067 = 1067:1067 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 109 dep = 1105:1105 arg1 0.41 1084:1087 0.41 1084:1087 0.41 1084:1087 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 109 dep = 1105:1105 arg1 CI 1090:1091 CI 1090:1091 CI 1090:1091 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 109 dep = 1105:1105 arg1 0.5-0.9 1056:1062 0.5-0.9 1056:1062 0.5-0.9 1056:1062 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 109 dep = 1105:1105 arg1 CD 1075:1076 CD 1075:1076 CD 1075:1076 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 109 dep = 1105:1105 arg1 0.3-0.6 1094:1100 0.3-0.6 1094:1100 0.3-0.6 1094:1100 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 11 110 theme therapeutic 1967:1977 arg1 efficacy 1979:1986 monoclonal antibody therapeutic efficacy 1947:1986 monoclonal antibody therapeutic efficacy 1947:1986 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 12 111 theme IBD 2041:2043 arg1 potential 2028:2036 translational potential 2014:2036 translational potential 2014:2036 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 12 111 theme IBD 2041:2043 arg1 biomarkers 2045:2054 IBD biomarkers 2041:2054 IBD biomarkers 2041:2054 IgG glycan profiles have translational potential as IBD biomarkers. 25895110 5 112 theme = 690:690 arg1 controls 678:685 controls 678:685 controls (n = 320) 678:695 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 5 112 theme = 690:690 arg1 n 688:688 n = 320 688:694 n = 320 688:694 METHODS IgG glycome composition in patients with UC (n = 507), CD (n = 287), and controls (n = 320) was analyzed by ultra performance liquid chromatography. 25895110 1 113 theme many 225:228 arg1 mechanisms 242:251 many etiological mechanisms 225:251 many etiological mechanisms described in IBD 225:268 BACKGROUND Glycobiology is an underexplored research area in inflammatory bowel disease (IBD), and glycans are relevant to many etiological mechanisms described in IBD. 25895110 9 114 from structures 1640:1649 arg1 absence 1590:1596 absence 1590:1596 absence of bisecting GlcNAc in monogalactosylated structures 1590:1649 The ratio of the presence to absence of bisecting GlcNAc in monogalactosylated structures was increased in patients with UC undergoing colectomy compared with no colectomy (FDR-adjusted, P = 0.05). 25895110 11 115 theme monoclonal 1947:1956 arg1 efficacy 1979:1986 monoclonal antibody therapeutic efficacy 1947:1986 monoclonal antibody therapeutic efficacy 1947:1986 Changes in IgG glycosylation may contribute to IBD pathogenesis and could alter monoclonal antibody therapeutic efficacy. 25895110 7 116 dep digalactosylation 979:995 arg1 odds 1002:1005 odds 1002:1005 odds ratio [OR] = 0.71; 95% confidence interval [CI 1002:1052 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 25895110 7 116 dep digalactosylation 979:995 arg1 UC 998:999 UC 998:999 UC 998:999 Both UC and CD were associated with significantly decreased IgG galactosylation (digalactosylation, UC: odds ratio [OR] = 0.71; 95% confidence interval [CI], 0.5-0.9; P = 0.01; CD: OR = 0.41; CI, 0.3-0.6; P = 1.4 × 10) and significant decrease in the proportion of sialylated structures in CD (OR = 0.46, CI, 0.3-0.6, P = 8.4 × 10). 26307003 6 0 theme simulations 1300:1310 arg1 analysis 1255:1262 detailed analysis 1246:1262 detailed analysis of the NMR structures and molecular simulations 1246:1310 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 5 1 theme binding 1214:1220 arg1 affinity 1222:1229 binding affinity 1214:1229 binding affinity 1214:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 4 2 theme cellulose 1052:1060 arg1 surface 1062:1068 the cellulose surface 1048:1068 the cellulose surface 1048:1068 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 3 3 theme Trichoderma 587:597 arg1 CBM 615:617 a Trichoderma reesei family 1 CBM 585:617 a Trichoderma reesei family 1 CBM 585:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 4 gly glycosylated 655:666 arg1 CBM 677:679 a glycosylated family 1 CBM 653:679 a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 653:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 6 5 theme NMR 1271:1273 arg1 structures 1275:1284 the NMR structures 1267:1284 the NMR structures 1267:1284 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 2 6 theme family 303:308 arg1 CBMs 312:315 family 1 CBMs 303:315 family 1 CBMs 303:315 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 3 7 attach attached 702:709 arg1 Thr1 719:722 both Thr1 714:722 both Thr1 714:722 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 7 attach attached 702:709 arg2 group 696:700 a mannose group 686:700 a mannose group attached to both Thr1 686:722 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 8 theme CBM 615:617 arg1 glycoforms 571:580 two glycoforms 567:580 two glycoforms of a Trichoderma reesei family 1 CBM 567:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 9 theme plant 211:215 arg1 polysaccharides 227:241 plant cell wall polysaccharides 211:241 plant cell wall polysaccharides 211:241 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 10 theme important 1644:1652 arg1 monosaccharides 1610:1624 monosaccharides 1610:1624 monosaccharides 1610:1624 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 10 theme important 1644:1652 arg1 modifications 1673:1685 likely important post-translational modifications 1637:1685 likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes 1637:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 3 11 theme family 746:751 arg1 CBM 755:757 a second family 1 CBM 737:757 a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14 737:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 6 12 theme CBM 1355:1357 arg1 backbone 1339:1346 the protein backbone 1327:1346 the protein backbone of the CBM 1327:1357 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 3 13 theme family 606:611 arg1 CBM 615:617 a Trichoderma reesei family 1 CBM 585:617 a Trichoderma reesei family 1 CBM 585:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 14 with CBM 677:679 arg1 groups 779:784 single mannose groups 764:784 single mannose groups attached to Thr1, Ser3 and Ser14 764:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 14 with CBM 677:679 arg1 group 696:700 a mannose group 686:700 a mannose group attached to both Thr1 686:722 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 14 with CBM 677:679 arg1 Ser3 728:731 Ser3 728:731 Ser3 728:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 15 theme wall 222:225 arg1 polysaccharides 227:241 plant cell wall polysaccharides 211:241 plant cell wall polysaccharides 211:241 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 4 16 theme additional 919:928 arg1 platforms 948:956 additional cellulose binding platforms 919:956 additional cellulose binding platforms 919:956 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 2 17 theme proteolytic 408:418 arg1 stability 420:428 thermal and proteolytic stability 396:428 stability 420:428 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 3 18 theme NMR 635:637 arg1 spectroscopy 639:650 NMR spectroscopy 635:650 NMR spectroscopy 635:650 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 19 theme single 764:769 arg1 groups 779:784 single mannose groups 764:784 single mannose groups attached to Thr1, Ser3 and Ser14 764:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 6 20 theme protein 1331:1337 arg1 backbone 1339:1346 the protein backbone 1327:1346 the protein backbone of the CBM 1327:1357 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 4 21 theme family 897:902 arg1 CBMs 906:909 family 1 CBMs 897:909 family 1 CBMs 897:909 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 4 22 from Ser14 888:892 arg1 monosaccharides 855:869 monosaccharides 855:869 monosaccharides at both Ser3 and Ser14 on family 1 CBMs 855:909 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 3 23 theme glycosylated 655:666 arg1 CBM 677:679 a glycosylated family 1 CBM 653:679 a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 653:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 24 theme ubiquitous 145:154 arg1 modules 126:132 UNLABELLED Family 1 carbohydrate-binding modules 85:132 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) 85:139 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 24 theme ubiquitous 145:154 arg1 components 156:165 ubiquitous components 145:165 ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose 145:276 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 25 theme enzymes 1750:1756 arg1 subdomains 1703:1712 these common subdomains 1690:1712 these common subdomains of fungal plant cell wall degrading enzymes 1690:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 25 theme enzymes 1750:1756 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 26 theme covalent 1587:1594 arg1 attachment 1596:1605 covalent attachment 1587:1605 covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes 1587:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 8 27 dep BMRB 1885:1888 arg1 25331 1897:1901 25331 1897:1901 25331 1897:1901 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 27 dep BMRB 1885:1888 arg1 codes 1890:1894 codes 1890:1894 codes 1890:1894 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 27 dep BMRB 1885:1888 arg1 25332 1907:1911 25332 1907:1911 25332 1907:1911 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 5 28 with agreement 1092:1100 arg1 work 1129:1132 previous experimental work 1107:1132 previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity 1107:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 6 29 theme monosaccharides 1405:1419 arg1 attachment 1391:1400 attachment 1391:1400 attachment of monosaccharides 1391:1419 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 5 30 theme experimental 1116:1127 arg1 work 1129:1132 previous experimental work 1107:1132 previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity 1107:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 8 31 theme Protein 1816:1822 arg1 Bank 1829:1832 the RCSB Protein Data Bank 1807:1832 the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) 1807:1859 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 7 32 theme wall 1735:1738 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 4 33 theme aromatic 989:996 arg1 residues 998:1005 well-characterized aromatic residues 970:1005 well-characterized aromatic residues at the binding interface, which align to the cellulose surface 970:1068 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 1 34 theme fungal 183:188 arg1 enzymes 190:196 multimodular fungal enzymes 170:196 multimodular fungal enzymes 170:196 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 35 theme plant 1724:1728 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 4 36 theme binding 940:946 arg1 platforms 948:956 additional cellulose binding platforms 919:956 additional cellulose binding platforms 919:956 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 2 37 theme CBMs 312:315 arg1 glycosylation 286:298 Native glycosylation 279:298 Native glycosylation of family 1 CBMs 279:315 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 38 theme cellulose 434:442 arg1 affinity 452:459 cellulose binding affinity 434:459 cellulose binding affinity 434:459 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 1 39 theme Family 96:101 arg1 CBMs 135:138 CBMs 135:138 CBMs 135:138 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 39 theme Family 96:101 arg1 modules 126:132 UNLABELLED Family 1 carbohydrate-binding modules 85:132 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) 85:139 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 39 theme Family 96:101 arg1 components 156:165 ubiquitous components 145:165 ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose 145:276 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 40 theme common 1696:1701 arg1 subdomains 1703:1712 these common subdomains 1690:1712 these common subdomains of fungal plant cell wall degrading enzymes 1690:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 40 theme common 1696:1701 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 0 41 theme O-glycosylation 0:14 arg1 effects 16:22 O-glycosylation effects 0:22 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 0:83 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 3 42 with CBM 755:757 arg1 group 696:700 a mannose group 686:700 a mannose group attached to both Thr1 686:722 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 42 with CBM 755:757 arg1 groups 779:784 single mannose groups 764:784 single mannose groups attached to Thr1, Ser3 and Ser14 764:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 42 with CBM 755:757 arg1 Ser3 728:731 Ser3 728:731 Ser3 728:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 43 theme carbohydrate-binding 105:124 arg1 CBMs 135:138 CBMs 135:138 CBMs 135:138 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 43 theme carbohydrate-binding 105:124 arg1 modules 126:132 UNLABELLED Family 1 carbohydrate-binding modules 85:132 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) 85:139 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 43 theme carbohydrate-binding 105:124 arg1 components 156:165 ubiquitous components 145:165 ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose 145:276 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 5 44 theme significant 1186:1196 arg1 improvements 1198:1209 significant improvements 1186:1209 significant improvements in binding affinity 1186:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 7 45 theme post-translational 1654:1671 arg1 monosaccharides 1610:1624 monosaccharides 1610:1624 monosaccharides 1610:1624 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 45 theme post-translational 1654:1671 arg1 modifications 1673:1685 likely important post-translational modifications 1637:1685 likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes 1637:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 46 theme present 1521:1527 arg1 study 1529:1533 the present study 1517:1533 the present study 1517:1533 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 4 47 theme binding 1014:1020 arg1 interface 1022:1030 the binding interface 1010:1030 the binding interface 1010:1030 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 3 48 from changes 498:504 arg1 properties 513:522 CBM properties 509:522 CBM properties 509:522 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 0 49 theme module 57:62 arg1 structures 73:82 family 1 carbohydrate-binding module solution structures 27:82 family 1 carbohydrate-binding module solution structures 27:82 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 4 50 from Ser3 879:882 arg1 monosaccharides 855:869 monosaccharides 855:869 monosaccharides at both Ser3 and Ser14 on family 1 CBMs 855:909 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 5 51 from improvements 1198:1209 arg1 affinity 1222:1229 binding affinity 1214:1229 binding affinity 1214:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 2 52 theme multiple 356:363 arg1 affinity 452:459 cellulose binding affinity 434:459 cellulose binding affinity 434:459 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 52 theme multiple 356:363 arg1 properties 374:383 multiple physical properties 356:383 multiple physical properties 356:383 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 52 theme multiple 356:363 arg1 stability 420:428 thermal and proteolytic stability 396:428 stability 420:428 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 3 53 theme mannose 688:694 arg1 group 696:700 a mannose group 686:700 a mannose group attached to both Thr1 686:722 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 8 54 theme BioMagRes 1869:1877 arg1 Bank 1879:1882 the BioMagRes Bank 1865:1882 the BioMagRes Bank (BMRB codes: 25331 and 25332) 1865:1912 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 55 dep Bank 1879:1882 arg1 BMRB 1885:1888 BMRB 1885:1888 BMRB codes: 25331 and 25332 1885:1911 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 5 56 from Ser3 1164:1167 arg1 glycans 1153:1159 glycans 1153:1159 glycans at Ser3 and Ser14 1153:1177 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 3 57 theme solution 544:551 arg1 structures 553:562 solution structures 544:562 solution structures of two glycoforms of a Trichoderma reesei family 1 CBM 544:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 6 58 theme molecular 1290:1298 arg1 simulations 1300:1310 molecular simulations 1290:1310 molecular simulations 1290:1310 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 3 59 theme second 739:744 arg1 CBM 755:757 a second family 1 CBM 737:757 a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14 737:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 6 60 theme structures 1275:1284 arg1 analysis 1255:1262 detailed analysis 1246:1262 detailed analysis of the NMR structures and molecular simulations 1246:1310 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 3 61 theme glycoforms 571:580 arg1 structures 553:562 solution structures 544:562 solution structures of two glycoforms of a Trichoderma reesei family 1 CBM 544:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 62 theme enzymes 190:196 arg1 modules 126:132 UNLABELLED Family 1 carbohydrate-binding modules 85:132 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) 85:139 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 62 theme enzymes 190:196 arg1 components 156:165 ubiquitous components 145:165 ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose 145:276 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 2 63 gly glycosylation 286:298 arg1 CBMs 312:315 family 1 CBMs 303:315 family 1 CBMs 303:315 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 5 64 from Ser14 1173:1177 arg1 glycans 1153:1159 glycans 1153:1159 glycans at Ser3 and Ser14 1153:1177 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 8 65 dep PDB 1835:1837 arg1 2MWK 1855:1858 2MWK 1855:1858 2MWK 1855:1858 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 65 dep PDB 1835:1837 arg1 2MWJ 1846:1849 2MWJ 1846:1849 2MWJ 1846:1849 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 65 dep PDB 1835:1837 arg1 codes 1839:1843 codes 1839:1843 codes 1839:1843 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 3 66 theme reesei 599:604 arg1 CBM 615:617 a Trichoderma reesei family 1 CBM 585:617 a Trichoderma reesei family 1 CBM 585:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 1 67 theme cell 217:220 arg1 polysaccharides 227:241 plant cell wall polysaccharides 211:241 plant cell wall polysaccharides 211:241 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 8 68 dep Bank 1829:1832 arg1 PDB 1835:1837 PDB 1835:1837 PDB codes: 2MWJ and 2MWK 1835:1858 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 8 69 theme DATABASE 1759:1766 arg1 data 1779:1782 DATABASE Structural data 1759:1782 DATABASE Structural data 1759:1782 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 7 70 theme monosaccharides 1610:1624 arg1 attachment 1596:1605 covalent attachment 1587:1605 covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes 1587:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 6 71 theme detailed 1246:1253 arg1 analysis 1255:1262 detailed analysis 1246:1262 detailed analysis of the NMR structures and molecular simulations 1246:1310 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 3 72 theme mannose 771:777 arg1 groups 779:784 single mannose groups 764:784 single mannose groups attached to Thr1, Ser3 and Ser14 764:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 2 73 theme Native 279:284 arg1 glycosylation 286:298 Native glycosylation 279:298 Native glycosylation of family 1 CBMs 279:315 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 4 74 theme similar 959:965 arg1 platforms 948:956 additional cellulose binding platforms 919:956 additional cellulose binding platforms 919:956 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 6 75 attach attached 1443:1450 arg1 Ser14 1455:1459 Ser14 1455:1459 Ser14 1455:1459 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 6 75 attach attached 1443:1450 arg2 mannose 1435:1441 the mannose 1431:1441 the mannose attached to Ser14 1431:1459 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 6 75 attach attached 1443:1450 arg2 flexible 1473:1480 flexible 1473:1480 flexible 1473:1480 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 8 76 theme RCSB 1811:1814 arg1 Bank 1829:1832 the RCSB Protein Data Bank 1807:1832 the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) 1807:1859 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 1 77 theme multimodular 170:181 arg1 enzymes 190:196 multimodular fungal enzymes 170:196 multimodular fungal enzymes 170:196 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 78 theme degrading 1740:1748 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 79 theme CBM 1556:1558 arg1 structures 1560:1569 family 1 CBM structures 1547:1569 family 1 CBM structures 1547:1569 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 4 80 theme well-characterized 970:987 arg1 residues 998:1005 well-characterized aromatic residues 970:1005 well-characterized aromatic residues at the binding interface, which align to the cellulose surface 970:1068 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 0 81 from effects 16:22 arg1 structures 73:82 family 1 carbohydrate-binding module solution structures 27:82 family 1 carbohydrate-binding module solution structures 27:82 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 3 82 gly glycoforms 571:580 arg1 CBM 615:617 a Trichoderma reesei family 1 CBM 585:617 a Trichoderma reesei family 1 CBM 585:617 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 4 83 from monosaccharides 855:869 arg1 CBMs 906:909 family 1 CBMs 897:909 family 1 CBMs 897:909 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 8 84 theme Data 1824:1827 arg1 Bank 1829:1832 the RCSB Protein Data Bank 1807:1832 the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) 1807:1859 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 7 85 theme cell 1730:1733 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 86 theme family 1547:1552 arg1 structures 1560:1569 family 1 CBM structures 1547:1569 family 1 CBM structures 1547:1569 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 4 87 theme cellulose 930:938 arg1 platforms 948:956 additional cellulose binding platforms 919:956 additional cellulose binding platforms 919:956 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 8 88 theme Structural 1768:1777 arg1 data 1779:1782 DATABASE Structural data 1759:1782 DATABASE Structural data 1759:1782 DATABASE Structural data have been deposited in the RCSB Protein Data Bank (PDB codes: 2MWJ and 2MWK) and the BioMagRes Bank (BMRB codes: 25331 and 25332) for CBM_M2 and CBM_M3, respectively. 26307003 1 89 theme UNLABELLED 85:94 arg1 CBMs 135:138 CBMs 135:138 CBMs 135:138 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 89 theme UNLABELLED 85:94 arg1 modules 126:132 UNLABELLED Family 1 carbohydrate-binding modules 85:132 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) 85:139 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 1 89 theme UNLABELLED 85:94 arg1 components 156:165 ubiquitous components 145:165 ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose 145:276 UNLABELLED Family 1 carbohydrate-binding modules (CBMs) are ubiquitous components of multimodular fungal enzymes that degrade plant cell wall polysaccharides and bind specifically to cellulose. 26307003 7 90 theme fungal 1717:1722 arg1 enzymes 1750:1756 fungal plant cell wall degrading enzymes 1717:1756 fungal plant cell wall degrading enzymes 1717:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 5 91 theme previous 1107:1114 arg1 work 1129:1132 previous experimental work 1107:1132 previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity 1107:1229 These results are in agreement with previous experimental work demonstrating that glycans at Ser3 and Ser14 impart significant improvements in binding affinity. 26307003 7 92 theme subdomains 1703:1712 arg1 monosaccharides 1610:1624 monosaccharides 1610:1624 monosaccharides 1610:1624 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 7 92 theme subdomains 1703:1712 arg1 modifications 1673:1685 likely important post-translational modifications 1637:1685 likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes 1637:1756 Overall, the present study reveals how family 1 CBM structures are affected by covalent attachment of monosaccharides, which are likely important post-translational modifications of these common subdomains of fungal plant cell wall degrading enzymes. 26307003 3 93 theme molecular 470:478 arg1 insights 480:487 molecular insights 470:487 molecular insights into the changes in CBM properties upon glycosylation 470:541 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 0 94 theme family 27:32 arg1 structures 73:82 family 1 carbohydrate-binding module solution structures 27:82 family 1 carbohydrate-binding module solution structures 27:82 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 3 95 attach attached 786:793 arg1 Ser14 813:817 Ser14 813:817 Ser14 813:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 95 attach attached 786:793 arg2 groups 779:784 single mannose groups 764:784 single mannose groups attached to Thr1, Ser3 and Ser14 764:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 95 attach attached 786:793 arg1 Thr1 798:801 Thr1 798:801 Thr1 798:801 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 95 attach attached 786:793 arg1 Ser3 804:807 Ser3 804:807 Ser3 804:807 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 96 theme CBM 509:511 arg1 properties 513:522 CBM properties 509:522 CBM properties 509:522 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 0 97 theme carbohydrate-binding 36:55 arg1 structures 73:82 family 1 carbohydrate-binding module solution structures 27:82 family 1 carbohydrate-binding module solution structures 27:82 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 4 98 from interface 1022:1030 arg1 residues 998:1005 well-characterized aromatic residues 970:1005 well-characterized aromatic residues at the binding interface, which align to the cellulose surface 970:1068 The structures clearly reveal that monosaccharides at both Ser3 and Ser14 on family 1 CBMs present additional cellulose binding platforms, similar to well-characterized aromatic residues at the binding interface, which align to the cellulose surface. 26307003 3 99 dep gain 465:468 arg1 CBM 677:679 a glycosylated family 1 CBM 653:679 a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 653:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 99 dep gain 465:468 arg1 CBM 755:757 a second family 1 CBM 737:757 a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14 737:817 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 3 100 theme family 668:673 arg1 CBM 677:679 a glycosylated family 1 CBM 653:679 a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 653:731 To gain molecular insights into the changes in CBM properties upon glycosylation, solution structures of two glycoforms of a Trichoderma reesei family 1 CBM were studied by NMR spectroscopy: a glycosylated family 1 CBM with a mannose group attached to both Thr1 and Ser3 and a second family 1 CBM with single mannose groups attached to Thr1, Ser3 and Ser14. 26307003 2 101 theme binding 444:450 arg1 affinity 452:459 cellulose binding affinity 434:459 cellulose binding affinity 434:459 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 0 102 theme solution 64:71 arg1 structures 73:82 family 1 carbohydrate-binding module solution structures 27:82 family 1 carbohydrate-binding module solution structures 27:82 O-glycosylation effects on family 1 carbohydrate-binding module solution structures. 26307003 6 103 from Ser3 1502:1505 arg1 mannose 1491:1497 the mannose 1487:1497 the mannose at Ser3 1487:1505 Additionally, detailed analysis of the NMR structures and molecular simulations indicates that the protein backbone of the CBM is not significantly altered by attachment of monosaccharides, and that the mannose attached to Ser14 may be more flexible than the mannose at Ser3. 26307003 2 104 theme physical 365:372 arg1 affinity 452:459 cellulose binding affinity 434:459 cellulose binding affinity 434:459 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 104 theme physical 365:372 arg1 properties 374:383 multiple physical properties 356:383 multiple physical properties 356:383 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 104 theme physical 365:372 arg1 stability 420:428 thermal and proteolytic stability 396:428 stability 420:428 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 26307003 2 105 theme thermal 396:402 arg1 stability 420:428 thermal and proteolytic stability 396:428 stability 420:428 Native glycosylation of family 1 CBMs has been shown to substantially impact multiple physical properties, including thermal and proteolytic stability and cellulose binding affinity. 27094473 10 0 theme glycan 1693:1698 arg1 structures 1700:1709 glycan structures 1693:1709 glycan structures 1693:1709 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 7 1 theme cell 1170:1173 arg1 proteomes 1180:1188 altered cell wall proteomes 1162:1188 altered cell wall proteomes 1162:1188 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 10 2 theme shorter 1728:1734 arg1 structures 1743:1752 shorter glycan structures 1728:1752 shorter glycan structures 1728:1752 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 1 3 theme profound 131:138 arg1 impacts 140:146 profound impacts 131:146 profound impacts 131:146 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 7 4 theme glycan 1141:1146 arg1 occupancy 1148:1156 decreased glycan occupancy 1131:1156 decreased glycan occupancy 1131:1156 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 8 5 theme earlier 1234:1240 arg1 mannosyltransferase 1242:1260 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 mannosyltransferase 1242:1260 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 8 6 theme hypoglycosylation 1327:1343 arg1 phenotypes 1345:1354 stronger hypoglycosylation phenotypes 1318:1354 stronger hypoglycosylation phenotypes 1318:1354 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 8 7 contain had 1314:1316 arg2 phenotypes 1345:1354 stronger hypoglycosylation phenotypes 1318:1354 stronger hypoglycosylation phenotypes 1318:1354 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 8 7 contain had 1314:1316 arg1 Mutations 1221:1229 Mutations 1221:1229 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis 1221:1312 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 4 8 with strains 572:578 arg1 deletions 585:593 deletions 585:593 deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 585:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 8 9 theme glucosyltransferase 1361:1379 arg1 defects 1381:1387 glucosyltransferase defects 1361:1387 glucosyltransferase defects 1361:1387 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 1 10 contain have 126:129 arg1 microheterogeneity 107:124 microheterogeneity 107:124 microheterogeneity 107:124 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 1 10 contain have 126:129 arg2 impacts 140:146 profound impacts 131:146 profound impacts 131:146 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 1 10 contain have 126:129 arg1 macro- 96:101 Glycan macro- 89:101 Glycan macro- 89:101 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 4 11 dep mannosyltransferases 643:662 arg1 Alg9 671:674 Alg9 671:674 Alg9 671:674 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 11 dep mannosyltransferases 643:662 arg1 Alg12 681:685 Alg12 681:685 Alg12 681:685 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 11 dep mannosyltransferases 643:662 arg1 Alg3 665:668 Alg3 665:668 Alg3 665:668 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 11 dep mannosyltransferases 643:662 arg1 mannosyltransferases 643:662 Endoplasmic Reticulum lumenal mannosyltransferases 613:662 Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12) 613:686 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 7 12 theme wild-type 1204:1212 arg1 cells 1214:1218 wild-type cells 1204:1218 wild-type cells 1204:1218 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 5 13 theme mature 868:873 arg1 proteins 875:882 mature proteins 868:882 mature proteins caused by these mutations 868:908 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 8 14 theme glycan 1294:1299 arg1 biosynthesis 1301:1312 glycan biosynthesis 1294:1312 glycan biosynthesis 1294:1312 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 11 15 theme site-specific 2009:2021 arg1 glycosylation 2023:2035 site-specific glycosylation 2009:2035 site-specific glycosylation 2009:2035 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 2 16 theme environmental 237:249 arg1 factors 251:257 physiological or environmental factors 220:257 physiological or environmental factors 220:257 This heterogeneity can be regulated by physiological or environmental factors. 27094473 5 17 theme spectrometry 936:947 arg1 workflow 965:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 4 18 from mutations 454:462 arg1 pathway 487:493 the N-glycosylation pathway 467:493 the N-glycosylation pathway 467:493 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 1 19 theme protein 151:157 arg1 folding 159:165 protein folding 151:165 protein folding 151:165 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 6 20 theme relative 1058:1065 arg1 abundance 1075:1083 relative protein abundance 1058:1083 relative protein abundance 1058:1083 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 11 21 theme quantitative 1920:1931 arg1 glycoproteomics 1933:1947 facile relative quantitative glycoproteomics 1904:1947 facile relative quantitative glycoproteomics 1904:1947 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 0 22 theme Glycosylation 64:76 arg1 Machinery 78:86 the Glycosylation Machinery 60:86 the Glycosylation Machinery 60:86 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 3 23 theme glycosylation 337:349 arg1 process 351:357 the glycosylation process 333:357 the glycosylation process 333:357 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 6 24 theme wall 1034:1037 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 6 25 theme mature 1022:1027 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 3 26 theme growing 367:373 arg1 number 375:380 a growing number 365:380 a growing number of Congenital Disorders of Glycosylation 365:421 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 10 27 theme mannosidase 1819:1829 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 9 28 theme mannosyltransferase 1410:1428 arg1 mutants 1430:1436 ER mannosyltransferase mutants 1407:1436 ER mannosyltransferase mutants 1407:1436 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 9 29 from changes 1467:1473 arg1 microheterogeneity 1485:1502 glycan microheterogeneity 1478:1502 glycan microheterogeneity 1478:1502 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 1 30 theme Glycan 89:94 arg1 macro- 96:101 Glycan macro- 89:101 Glycan macro- 89:101 Glycan macro- and microheterogeneity have profound impacts on protein folding and function. 27094473 3 31 theme Glycosylation 409:421 arg1 Disorders 396:404 Congenital Disorders 385:404 Congenital Disorders of Glycosylation 385:421 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 10 32 theme ER 1596:1597 arg1 glucosyltransferase 1599:1617 ER glucosyltransferase and oligosaccharyltransferase subunit mutants 1596:1663 glucosyltransferase 1599:1617 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 5 33 dep developed 913:921 arg1 measure 805:811 measure 805:811 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations 802:908 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 10 34 theme quality 1803:1809 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 0 35 theme SWATH-MS 0:7 arg1 Glycoproteomics 9:23 SWATH-MS Glycoproteomics 0:23 SWATH-MS Glycoproteomics 0:23 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 10 36 contain had 1724:1726 arg2 structures 1743:1752 shorter glycan structures 1728:1752 shorter glycan structures 1728:1752 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 10 36 contain had 1724:1726 arg1 cells 1718:1722 ost3Δ cells 1712:1722 ost3Δ cells 1712:1722 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 6 37 theme different 1093:1101 arg1 mutants 1103:1109 the different mutants 1089:1109 the different mutants 1089:1109 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 3 38 theme Disorders 396:404 arg1 number 375:380 a growing number 365:380 a growing number of Congenital Disorders of Glycosylation 365:421 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 9 39 theme global 1460:1465 arg1 changes 1467:1473 substantial global changes 1448:1473 substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains 1448:1584 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 4 40 theme Endoplasmic 613:623 arg1 Alg9 671:674 Alg9 671:674 Alg9 671:674 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 40 theme Endoplasmic 613:623 arg1 Alg12 681:685 Alg12 681:685 Alg12 681:685 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 40 theme Endoplasmic 613:623 arg1 Alg3 665:668 Alg3 665:668 Alg3 665:668 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 40 theme Endoplasmic 613:623 arg1 mannosyltransferases 643:662 Endoplasmic Reticulum lumenal mannosyltransferases 613:662 Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12) 613:686 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 10 41 theme subunit 1649:1655 arg1 mutants 1657:1663 ER glucosyltransferase and oligosaccharyltransferase subunit mutants 1596:1663 mutants 1657:1663 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 4 42 from deletions 585:593 arg1 genes 598:602 genes 598:602 genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 598:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 6 43 gly glycoproteins 1039:1051 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 6 44 from occupancy 1009:1017 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 0 45 from Consequences 33:44 arg1 Machinery 78:86 the Glycosylation Machinery 60:86 the Glycosylation Machinery 60:86 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 8 46 from Mutations 1221:1229 arg1 mannosyltransferase 1242:1260 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 mannosyltransferase 1242:1260 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 8 46 from Mutations 1221:1229 arg1 steps 1285:1289 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 steps 1285:1289 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 2 47 theme physiological 220:232 arg1 factors 251:257 physiological or environmental factors 220:257 physiological or environmental factors 220:257 This heterogeneity can be regulated by physiological or environmental factors. 27094473 9 48 from truncations 1520:1530 arg1 glycan 1539:1544 the glycan 1535:1544 the glycan transferred to protein in these strains 1535:1584 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 10 49 with consistent 1769:1778 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 10 50 theme ost3Δ 1712:1716 arg1 cells 1718:1722 ost3Δ cells 1712:1722 ost3Δ cells 1712:1722 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 7 51 theme altered 1162:1168 arg1 proteomes 1180:1188 altered cell wall proteomes 1162:1188 altered cell wall proteomes 1162:1188 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 4 52 theme lumenal 635:641 arg1 Alg9 671:674 Alg9 671:674 Alg9 671:674 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 52 theme lumenal 635:641 arg1 Alg12 681:685 Alg12 681:685 Alg12 681:685 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 52 theme lumenal 635:641 arg1 Alg3 665:668 Alg3 665:668 Alg3 665:668 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 52 theme lumenal 635:641 arg1 mannosyltransferases 643:662 Endoplasmic Reticulum lumenal mannosyltransferases 613:662 Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12) 613:686 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 10 53 theme glycan 1736:1741 arg1 structures 1743:1752 shorter glycan structures 1728:1752 shorter glycan structures 1728:1752 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 9 54 with consistent 1504:1513 arg1 truncations 1520:1530 truncations 1520:1530 truncations in the glycan transferred to protein in these strains 1520:1584 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 7 55 theme decreased 1131:1139 arg1 occupancy 1148:1156 decreased glycan occupancy 1131:1156 decreased glycan occupancy 1131:1156 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 8 56 theme biosynthesis 1301:1312 arg1 mannosyltransferase 1242:1260 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 mannosyltransferase 1242:1260 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 8 56 theme biosynthesis 1301:1312 arg1 steps 1285:1289 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 steps 1285:1289 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 3 57 theme unregulated 269:279 arg1 heterogeneity 281:293 unregulated heterogeneity 269:293 unregulated heterogeneity 269:293 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 8 58 theme stronger 1318:1325 arg1 phenotypes 1345:1354 stronger hypoglycosylation phenotypes 1318:1354 stronger hypoglycosylation phenotypes 1318:1354 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 4 59 from defects 503:509 arg1 proteins 521:528 mature proteins 514:528 mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 514:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 10 60 theme increased 1785:1793 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 11 61 theme global 1988:1993 arg1 regulation 1995:2004 global regulation 1988:2004 global regulation of site-specific glycosylation 1988:2035 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 8 62 theme glucosyltransferase 1265:1283 arg1 steps 1285:1289 earlier mannosyltransferase or glucosyltransferase steps 1234:1289 steps 1285:1289 Mutations in earlier mannosyltransferase or glucosyltransferase steps of glycan biosynthesis had stronger hypoglycosylation phenotypes, but glucosyltransferase defects were more severe. 27094473 6 63 theme glycan 987:992 arg1 structures 994:1003 glycan structures 987:1003 glycan structures 987:1003 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 0 64 from Machinery 78:86 arg1 Consequences 33:44 Consequences 33:44 Consequences of Defects in the Glycosylation Machinery 33:86 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 10 65 gly hypoglycosylating 1859:1875 arg0 mutant 1877:1882 this severely hypoglycosylating mutant 1845:1882 this severely hypoglycosylating mutant 1845:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 5 66 theme SWATH-mass 925:934 arg1 workflow 965:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 11 67 theme relative 1911:1918 arg1 glycoproteomics 1933:1947 facile relative quantitative glycoproteomics 1904:1947 facile relative quantitative glycoproteomics 1904:1947 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 11 68 theme glycosylation 2023:2035 arg1 regulation 1995:2004 global regulation 1988:2004 global regulation of site-specific glycosylation 1988:2035 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 5 69 theme glycoproteomics 949:963 arg1 workflow 965:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 a SWATH-mass spectrometry glycoproteomics workflow 923:972 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 6 70 theme protein 1067:1073 arg1 abundance 1075:1083 relative protein abundance 1058:1083 relative protein abundance 1058:1083 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 4 71 theme N-glycosylation 471:485 arg1 pathway 487:493 the N-glycosylation pathway 467:493 the N-glycosylation pathway 467:493 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 72 theme oligosaccharyltransferase 743:767 arg1 Ost5 785:788 Ost5 785:788 Ost5 785:788 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 72 theme oligosaccharyltransferase 743:767 arg1 Ost6 795:798 Ost6 795:798 Ost6 795:798 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 72 theme oligosaccharyltransferase 743:767 arg1 Ost3 779:782 Ost3 779:782 Ost3 779:782 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 72 theme oligosaccharyltransferase 743:767 arg1 subunits 769:776 oligosaccharyltransferase subunits 743:776 oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 743:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 73 theme viable 540:545 arg1 strains 572:578 all viable Saccharomyces cerevisiae strains 536:578 all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 536:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 74 theme cerevisiae 561:570 arg1 strains 572:578 all viable Saccharomyces cerevisiae strains 536:578 all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 536:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 6 75 theme cell 1029:1032 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 3 76 theme Congenital 385:394 arg1 Disorders 396:404 Congenital Disorders 385:404 Congenital Disorders of Glycosylation 385:421 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 4 77 theme mature 514:519 arg1 proteins 521:528 mature proteins 514:528 mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 514:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 9 78 theme ER 1407:1408 arg1 mutants 1430:1436 ER mannosyltransferase mutants 1407:1436 ER mannosyltransferase mutants 1407:1436 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 10 79 from change 1683:1688 arg1 structures 1700:1709 glycan structures 1693:1709 glycan structures 1693:1709 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 0 80 from Defects 49:55 arg1 Machinery 78:86 the Glycosylation Machinery 60:86 the Glycosylation Machinery 60:86 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 10 81 theme control 1811:1817 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 4 82 dep subunits 769:776 arg1 Ost5 785:788 Ost5 785:788 Ost5 785:788 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 82 dep subunits 769:776 arg1 Ost6 795:798 Ost6 795:798 Ost6 795:798 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 82 dep subunits 769:776 arg1 Ost3 779:782 Ost3 779:782 Ost3 779:782 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 82 dep subunits 769:776 arg1 subunits 769:776 oligosaccharyltransferase subunits 743:776 oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6) 743:799 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 10 83 theme protein 1795:1801 arg1 processing 1831:1840 increased protein quality control mannosidase processing 1785:1840 increased protein quality control mannosidase processing in this severely hypoglycosylating mutant 1785:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 5 84 theme glycan 828:833 arg1 macro- 835:840 glycan macro- 828:840 glycan macro- 828:840 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 9 85 theme substantial 1448:1458 arg1 changes 1467:1473 substantial global changes 1448:1473 substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains 1448:1584 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 4 86 theme Reticulum 625:633 arg1 Alg9 671:674 Alg9 671:674 Alg9 671:674 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 86 theme Reticulum 625:633 arg1 Alg12 681:685 Alg12 681:685 Alg12 681:685 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 86 theme Reticulum 625:633 arg1 Alg3 665:668 Alg3 665:668 Alg3 665:668 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 86 theme Reticulum 625:633 arg1 mannosyltransferases 643:662 Endoplasmic Reticulum lumenal mannosyltransferases 613:662 Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12) 613:686 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 6 87 from structures 994:1003 arg1 glycoproteins 1039:1051 mature cell wall glycoproteins 1022:1051 mature cell wall glycoproteins 1022:1051 We measured glycan structures and occupancy on mature cell wall glycoproteins, and relative protein abundance, in the different mutants. 27094473 0 88 theme Defects 49:55 arg1 Consequences 33:44 Consequences 33:44 Consequences of Defects in the Glycosylation Machinery 33:86 SWATH-MS Glycoproteomics Reveals Consequences of Defects in the Glycosylation Machinery. 27094473 4 89 dep glucosyltransferases 689:708 arg1 Die2/Alg10 727:736 Die2/Alg10 727:736 Die2/Alg10 727:736 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 89 dep glucosyltransferases 689:708 arg1 Alg6 711:714 Alg6 711:714 Alg6 711:714 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 89 dep glucosyltransferases 689:708 arg1 glucosyltransferases 689:708 glucosyltransferases 689:708 glucosyltransferases (Alg6, Alg8, and Die2/Alg10) 689:737 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 4 89 dep glucosyltransferases 689:708 arg1 Alg8 717:720 Alg8 717:720 Alg8 717:720 We systematically studied how mutations in the N-glycosylation pathway lead to defects in mature proteins using all viable Saccharomyces cerevisiae strains with deletions in genes encoding Endoplasmic Reticulum lumenal mannosyltransferases (Alg3, Alg9, and Alg12), glucosyltransferases (Alg6, Alg8, and Die2/Alg10), or oligosaccharyltransferase subunits (Ost3, Ost5, and Ost6). 27094473 10 90 theme oligosaccharyltransferase 1623:1647 arg1 mutants 1657:1663 ER glucosyltransferase and oligosaccharyltransferase subunit mutants 1596:1663 mutants 1657:1663 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 7 91 theme wall 1175:1178 arg1 proteomes 1180:1188 altered cell wall proteomes 1162:1188 altered cell wall proteomes 1162:1188 All mutants showed decreased glycan occupancy and altered cell wall proteomes compared with wild-type cells. 27094473 9 92 theme glycan 1478:1483 arg1 microheterogeneity 1485:1502 glycan microheterogeneity 1478:1502 glycan microheterogeneity 1478:1502 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 5 93 from changes 817:823 arg1 microheterogeneity 846:863 microheterogeneity 846:863 microheterogeneity 846:863 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 5 93 from changes 817:823 arg1 proteins 875:882 mature proteins 868:882 mature proteins caused by these mutations 868:908 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 5 93 from changes 817:823 arg1 macro- 835:840 glycan macro- 828:840 glycan macro- 828:840 To measure the changes in glycan macro- and microheterogeneity in mature proteins caused by these mutations we developed a SWATH-mass spectrometry glycoproteomics workflow. 27094473 11 94 theme facile 1904:1909 arg1 glycoproteomics 1933:1947 facile relative quantitative glycoproteomics 1904:1947 facile relative quantitative glycoproteomics 1904:1947 This method allows facile relative quantitative glycoproteomics, and our results provide insights into global regulation of site-specific glycosylation. 27094473 9 95 theme consistent 1504:1513 arg1 changes 1467:1473 substantial global changes 1448:1473 substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains 1448:1584 ER mannosyltransferase mutants displayed substantial global changes in glycan microheterogeneity consistent with truncations in the glycan transferred to protein in these strains. 27094473 10 96 theme hypoglycosylating 1859:1875 arg1 mutant 1877:1882 this severely hypoglycosylating mutant 1845:1882 this severely hypoglycosylating mutant 1845:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27094473 3 97 from mutations 320:328 arg1 process 351:357 the glycosylation process 333:357 the glycosylation process 333:357 However, unregulated heterogeneity can lead to disease, and mutations in the glycosylation process cause a growing number of Congenital Disorders of Glycosylation. 27094473 10 98 from processing 1831:1840 arg1 mutant 1877:1882 this severely hypoglycosylating mutant 1845:1882 this severely hypoglycosylating mutant 1845:1882 Although ER glucosyltransferase and oligosaccharyltransferase subunit mutants broadly showed no change in glycan structures, ost3Δ cells had shorter glycan structures at some sites, consistent with increased protein quality control mannosidase processing in this severely hypoglycosylating mutant. 27412689 8 0 theme carbon 1611:1616 arg1 PGC-LC 1641:1646 PGC-LC 1641:1646 PGC-LC 1641:1646 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 8 0 theme carbon 1611:1616 arg1 chromatography 1625:1638 porous graphitized carbon liquid chromatography 1592:1638 porous graphitized carbon liquid chromatography (PGC-LC) 1592:1647 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 4 1 theme desorption/ionization 655:675 arg1 spectrometry 690:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry 633:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 7 2 theme FFPE 1445:1448 arg1 tissue 1465:1470 FFPE ovarian cancer tissue regions 1445:1478 FFPE ovarian cancer tissue regions 1445:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 4 3 theme MALDI 678:682 arg1 spectrometry 690:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry 633:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 11 4 theme tumor 2133:2137 arg1 region 2146:2151 the tumor tissue region 2129:2151 the tumor tissue region 2129:2151 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 7 5 theme peptide-N-glycosidase 1497:1517 arg1 F 1519:1519 enzymatic peptide-N-glycosidase F 1487:1519 enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans 1487:1551 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 7 5 theme peptide-N-glycosidase 1497:1517 arg1 F 1529:1529 PNGase F 1522:1529 PNGase F 1522:1529 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 12 6 theme non-tumor 2262:2270 arg1 tissue 2272:2277 non-tumor tissue 2262:2277 non-tumor tissue 2262:2277 Therefore, tumor and non-tumor tissue regions were clearly demarcated solely on their N-glycan structure distributions. 27412689 10 7 theme cancer 2046:2051 arg1 sections 2053:2060 the ovarian cancer sections 2034:2060 the ovarian cancer sections 2034:2060 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 2 8 theme glycoproteomic 267:280 arg1 studies 292:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 1 9 theme adult 190:194 arg1 women 196:200 adult women 190:200 adult women 190:200 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 6 10 theme high 1225:1228 arg1 images 1241:1246 high resolution images 1225:1246 high resolution images of the glycan structure distribution in ovarian cancer tissue 1225:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 2 11 theme protein 324:330 arg1 alterations 354:364 extensive protein glycosylation pattern alterations 314:364 extensive protein glycosylation pattern alterations in ovarian cancer 314:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 7 12 dep tissue 1465:1470 arg1 regions 1472:1478 regions 1472:1478 FFPE ovarian cancer tissue regions 1445:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 7 13 theme first 1431:1435 arg1 time 1437:1440 the first time 1427:1440 the first time in FFPE ovarian cancer tissue regions 1427:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 7 14 theme N-linked 1315:1322 arg1 repertoires 1331:1341 The N-linked glycan repertoires 1311:1341 The N-linked glycan repertoires carried by the proteins in these tissue regions 1311:1389 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 1 15 theme five-year 209:217 arg1 rate 236:239 a five-year overall survival rate 207:239 a five-year overall survival rate of only 30% 207:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 7 16 theme cancer 1458:1463 arg1 tissue 1465:1470 FFPE ovarian cancer tissue regions 1445:1478 FFPE ovarian cancer tissue regions 1445:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 12 17 theme tumor 2252:2256 arg1 regions 2279:2285 tumor and non-tumor tissue regions 2252:2285 tumor and non-tumor tissue regions 2252:2285 Therefore, tumor and non-tumor tissue regions were clearly demarcated solely on their N-glycan structure distributions. 27412689 9 18 theme PNGase 1905:1910 arg1 F 1912:1912 PNGase F 1905:1912 PNGase F 1905:1912 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 1 19 theme survival 227:234 arg1 rate 236:239 a five-year overall survival rate 207:239 a five-year overall survival rate of only 30% 207:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 11 20 theme intervening 2221:2231 arg1 stroma 2233:2238 the intervening stroma 2217:2238 the intervening stroma 2217:2238 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 6 21 theme cancer 1296:1301 arg1 tissue 1303:1308 ovarian cancer tissue 1288:1308 ovarian cancer tissue 1288:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 1 22 theme Ovarian 135:141 arg1 cancer 143:148 Ovarian cancer 135:148 Ovarian cancer 135:148 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 1 22 theme Ovarian 135:141 arg1 malignancy 176:185 a fatal gynaecological malignancy 153:185 a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30% 153:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 4 23 theme N-glycan 803:810 arg1 compositions 812:823 profile N-glycan compositions 795:823 profile N-glycan compositions released from proteins in tissue-specific regions 795:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 2 24 theme Glycomic 254:261 arg1 studies 292:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 5 25 theme FFPE 993:996 arg1 sections 1005:1012 FFPE tissue sections 993:1012 FFPE tissue sections of advanced serous ovarian cancers (n = 3) 993:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 9 26 theme N-glycan 1841:1848 arg1 distribution 1820:1831 distribution 1820:1831 distribution 1820:1831 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 9 26 theme N-glycan 1841:1848 arg1 location 1807:1814 location 1807:1814 location 1807:1814 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 6 27 theme structure 1262:1270 arg1 distribution 1272:1283 the glycan structure distribution 1251:1283 the glycan structure distribution in ovarian cancer tissue 1251:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 7 28 theme tissue 1376:1381 arg1 regions 1383:1389 these tissue regions 1370:1389 these tissue regions 1370:1389 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 4 29 theme tissue 760:765 arg1 sections 767:774 formalin-fixed paraffin-embedded (FFPE) tissue sections 720:774 formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions 720:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 6 30 from tissue 1303:1308 arg1 images 1241:1246 high resolution images 1225:1246 high resolution images of the glycan structure distribution in ovarian cancer tissue 1225:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 9 31 dep location 1807:1814 arg1 the 1803:1805 the 1803:1805 the 1803:1805 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 5 32 theme interest 909:916 arg1 regions 898:904 tissue regions 891:904 tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) 891:972 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 33 theme necrotic 958:965 arg1 areas 967:971 necrotic areas 958:971 necrotic areas 958:971 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 9 34 from distribution 1820:1831 arg1 sections 1873:1880 FFPE ovarian cancer sections 1853:1880 FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI 1853:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 9 35 theme FFPE 1853:1856 arg1 sections 1873:1880 FFPE ovarian cancer sections 1853:1880 FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI 1853:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 1 36 with malignancy 176:185 arg1 rate 236:239 a five-year overall survival rate 207:239 a five-year overall survival rate of only 30% 207:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 9 37 from location 1807:1814 arg1 sections 1873:1880 FFPE ovarian cancer sections 1853:1880 FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI 1853:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 9 38 theme cancer 1866:1871 arg1 sections 1873:1880 FFPE ovarian cancer sections 1853:1880 FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI 1853:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 4 39 theme novel 560:564 arg1 method 566:571 A novel method 558:571 A novel method for investigating tissue-specific N-linked glycans 558:622 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 4 40 theme tissue-specific 851:865 arg1 regions 867:873 tissue-specific regions 851:873 tissue-specific regions 851:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 6 41 from distribution 1272:1283 arg1 tissue 1303:1308 ovarian cancer tissue 1288:1308 ovarian cancer tissue 1288:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 5 42 theme ovarian 1033:1039 arg1 n 1050:1050 n = 3 1050:1054 n = 3 1050:1054 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 42 theme ovarian 1033:1039 arg1 cancers 1041:1047 advanced serous ovarian cancers 1017:1047 advanced serous ovarian cancers (n = 3) 1017:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 8 43 theme porous 1592:1597 arg1 PGC-LC 1641:1646 PGC-LC 1641:1646 PGC-LC 1641:1646 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 8 43 theme porous 1592:1597 arg1 chromatography 1625:1638 porous graphitized carbon liquid chromatography 1592:1638 porous graphitized carbon liquid chromatography (PGC-LC) 1592:1647 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 0 44 theme Cancer 119:124 arg1 Tissues 126:132 Ovarian Cancer Tissues 111:132 Ovarian Cancer Tissues 111:132 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 4 45 from proteins 839:846 arg1 regions 867:873 tissue-specific regions 851:873 tissue-specific regions 851:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 7 46 from time 1437:1440 arg1 tissue 1465:1470 FFPE ovarian cancer tissue regions 1445:1478 FFPE ovarian cancer tissue regions 1445:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 0 47 from Spectrometry 28:39 arg1 Tissue 77:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 10 48 theme sections 2053:2060 arg1 regions 2023:2029 particular regions 2012:2029 particular regions of the ovarian cancer sections 2012:2060 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 3 49 theme tissue-specific 469:483 arg1 changes 485:491 unearth tissue-specific changes 461:491 unearth tissue-specific changes that occur in the development and progression of ovarian cancer 461:555 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 0 50 theme N-glycan 0:7 arg1 Spectrometry 28:39 N-glycan MALDI Imaging Mass Spectrometry 0:39 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue 0:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 3 51 theme spatio-temporal 396:410 arg1 investigation 412:424 spatio-temporal investigation 396:424 spatio-temporal investigation of these glycosylation changes 396:455 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 0 52 theme Imaging 15:21 arg1 Spectrometry 28:39 N-glycan MALDI Imaging Mass Spectrometry 0:39 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue 0:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 5 53 theme advanced 1017:1024 arg1 n 1050:1050 n = 3 1050:1054 n = 3 1050:1054 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 53 theme advanced 1017:1024 arg1 cancers 1041:1047 advanced serous ovarian cancers 1017:1047 advanced serous ovarian cancers (n = 3) 1017:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 10 54 theme tissue-specific 1949:1963 arg1 distribution 1965:1976 A tissue-specific distribution 1947:1976 A tissue-specific distribution of N-glycan structures 1947:1999 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 9 55 theme high 1920:1923 arg1 MALDI-MSI 1936:1944 high resolution MALDI-MSI 1920:1944 high resolution MALDI-MSI 1920:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 3 56 theme glycosylation 435:447 arg1 changes 449:455 these glycosylation changes 429:455 these glycosylation changes 429:455 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 12 57 theme structure 2336:2344 arg1 distributions 2346:2358 their N-glycan structure distributions 2321:2358 their N-glycan structure distributions 2321:2358 Therefore, tumor and non-tumor tissue regions were clearly demarcated solely on their N-glycan structure distributions. 27412689 3 58 theme ovarian 542:548 arg1 cancer 550:555 ovarian cancer 542:555 ovarian cancer 542:555 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 0 59 theme Formalin-Fixed 44:57 arg1 Tissue 77:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 4 60 theme tissue-specific 591:605 arg1 glycans 616:622 tissue-specific N-linked glycans 591:622 tissue-specific N-linked glycans 591:622 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 10 61 theme structures 1990:1999 arg1 distribution 1965:1976 A tissue-specific distribution 1947:1976 A tissue-specific distribution of N-glycan structures 1947:1999 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 5 62 dep tumor 924:928 arg1 e.g. 919:922 e.g. 919:922 e.g. 919:922 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 6 63 theme resolution 1230:1239 arg1 images 1241:1246 high resolution images 1225:1246 high resolution images of the glycan structure distribution in ovarian cancer tissue 1225:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 5 64 theme = 1052:1052 arg1 n 1050:1050 n = 3 1050:1054 n = 3 1050:1054 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 64 theme = 1052:1052 arg1 cancers 1041:1047 advanced serous ovarian cancers 1017:1047 advanced serous ovarian cancers (n = 3) 1017:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 6 65 theme tissue-specific 1177:1191 arg1 glycans 1193:1199 the tissue-specific glycans 1173:1199 the tissue-specific glycans 1173:1199 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 8 66 theme negative 1684:1691 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 9 67 theme resolution 1925:1934 arg1 MALDI-MSI 1936:1944 high resolution MALDI-MSI 1920:1944 high resolution MALDI-MSI 1920:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 5 68 attach isolated 979:986 arg2 regions 898:904 tissue regions 891:904 tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) 891:972 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 68 attach isolated 979:986 arg1 sections 1005:1012 FFPE tissue sections 993:1012 FFPE tissue sections of advanced serous ovarian cancers (n = 3) 993:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 4 69 dep formalin-fixed 720:733 arg1 paraffin-embedded 735:751 paraffin-embedded 735:751 paraffin-embedded 735:751 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 4 70 theme spectrometry 690:701 arg1 MSI 712:714 MSI 712:714 MSI 712:714 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 4 70 theme spectrometry 690:701 arg1 imaging 703:709 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging 633:709 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 1 71 theme % 251:251 arg1 rate 236:239 a five-year overall survival rate 207:239 a five-year overall survival rate of only 30% 207:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 7 72 theme N-glycans 1543:1551 arg1 release 1532:1538 enzymatic peptide-N-glycosidase F (PNGase F) release 1487:1538 enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans 1487:1551 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 2 73 from alterations 354:364 arg1 cancer 377:382 ovarian cancer 369:382 ovarian cancer 369:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 6 74 theme structural 1147:1156 arg1 information 1158:1168 structural information 1147:1168 structural information on the tissue-specific glycans 1147:1199 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 4 75 theme laser 649:653 arg1 spectrometry 690:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry 633:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 10 76 theme particular 2012:2021 arg1 regions 2023:2029 particular regions 2012:2029 particular regions of the ovarian cancer sections 2012:2060 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 4 77 link N-linked 607:614 arg1 glycans 616:622 tissue-specific N-linked glycans 591:622 tissue-specific N-linked glycans 591:622 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 7 78 theme PNGase 1522:1527 arg1 F 1519:1519 enzymatic peptide-N-glycosidase F 1487:1519 enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans 1487:1551 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 7 78 theme PNGase 1522:1527 arg1 F 1529:1529 PNGase F 1522:1529 PNGase F 1522:1529 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 2 79 theme ovarian 369:375 arg1 cancer 377:382 ovarian cancer 369:382 ovarian cancer 369:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 2 80 theme profiling 282:290 arg1 studies 292:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies 254:298 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 7 81 theme F 1519:1519 arg1 release 1532:1538 enzymatic peptide-N-glycosidase F (PNGase F) release 1487:1538 enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans 1487:1551 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 4 82 theme formalin-fixed 720:733 arg1 sections 767:774 formalin-fixed paraffin-embedded (FFPE) tissue sections 720:774 formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions 720:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 10 83 theme ovarian 2038:2044 arg1 sections 2053:2060 the ovarian cancer sections 2034:2060 the ovarian cancer sections 2034:2060 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 2 84 theme glycosylation 332:344 arg1 alterations 354:364 extensive protein glycosylation pattern alterations 314:364 extensive protein glycosylation pattern alterations in ovarian cancer 314:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 5 85 theme adipose 939:945 arg1 tissue 947:952 adipose tissue 939:952 adipose tissue 939:952 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 7 86 theme enzymatic 1487:1495 arg1 F 1519:1519 enzymatic peptide-N-glycosidase F 1487:1519 enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans 1487:1551 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 7 86 theme enzymatic 1487:1495 arg1 F 1529:1529 PNGase F 1522:1529 PNGase F 1522:1529 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 12 87 theme tissue 2272:2277 arg1 regions 2279:2285 tumor and non-tumor tissue regions 2252:2285 tumor and non-tumor tissue regions 2252:2285 Therefore, tumor and non-tumor tissue regions were clearly demarcated solely on their N-glycan structure distributions. 27412689 12 88 theme N-glycan 2327:2334 arg1 distributions 2346:2358 their N-glycan structure distributions 2321:2358 their N-glycan structure distributions 2321:2358 Therefore, tumor and non-tumor tissue regions were clearly demarcated solely on their N-glycan structure distributions. 27412689 2 89 theme extensive 314:322 arg1 alterations 354:364 extensive protein glycosylation pattern alterations 314:364 extensive protein glycosylation pattern alterations in ovarian cancer 314:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 11 90 theme complex/hybrid 2161:2174 arg1 N-glycans 2176:2184 complex/hybrid N-glycans 2161:2184 complex/hybrid N-glycans 2161:2184 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 7 91 link N-linked 1315:1322 arg1 repertoires 1331:1341 The N-linked glycan repertoires 1311:1341 The N-linked glycan repertoires carried by the proteins in these tissue regions 1311:1389 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 1 92 theme overall 219:225 arg1 rate 236:239 a five-year overall survival rate 207:239 a five-year overall survival rate of only 30% 207:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 8 93 theme induced 1663:1669 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 4 94 attach released 825:832 arg2 compositions 812:823 profile N-glycan compositions 795:823 profile N-glycan compositions released from proteins in tissue-specific regions 795:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 4 94 attach released 825:832 arg1 proteins 839:846 proteins 839:846 proteins in tissue-specific regions 839:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 6 95 theme ovarian 1288:1294 arg1 tissue 1303:1308 ovarian cancer tissue 1288:1308 ovarian cancer tissue 1288:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 7 96 from proteins 1358:1365 arg1 regions 1383:1389 these tissue regions 1370:1389 these tissue regions 1370:1389 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 6 97 used used 1094:1097 arg2 MALDI-MSI 1079:1087 MALDI-MSI 1079:1087 MALDI-MSI 1079:1087 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 6 97 used used 1094:1097 arg2 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 6 97 used used 1094:1097 arg2 techniques 1116:1125 complementary techniques 1102:1125 complementary techniques 1102:1125 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 11 98 theme tissue 2139:2144 arg1 region 2146:2151 the tumor tissue region 2129:2151 the tumor tissue region 2129:2151 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 8 99 theme released 1558:1565 arg1 glycans 1567:1573 The released glycans 1554:1573 The released glycans 1554:1573 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 6 100 theme distribution 1272:1283 arg1 images 1241:1246 high resolution images 1225:1246 high resolution images of the glycan structure distribution in ovarian cancer tissue 1225:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 4 101 theme profile 795:801 arg1 compositions 812:823 profile N-glycan compositions 795:823 profile N-glycan compositions released from proteins in tissue-specific regions 795:873 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 11 102 theme mannose 2081:2087 arg1 glycans 2089:2095 high mannose glycans 2076:2095 high mannose glycans 2076:2095 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 6 103 theme glycan 1255:1260 arg1 distribution 1272:1283 the glycan structure distribution 1251:1283 the glycan structure distribution in ovarian cancer tissue 1251:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 8 104 theme mode 1693:1696 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 11 105 from stroma 2233:2238 arg1 abundant 2205:2212 abundant 2205:2212 abundant 2205:2212 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 1 106 theme gynaecological 161:174 arg1 cancer 143:148 Ovarian cancer 135:148 Ovarian cancer 135:148 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 1 106 theme gynaecological 161:174 arg1 malignancy 176:185 a fatal gynaecological malignancy 153:185 a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30% 153:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 9 107 theme ovarian 1858:1864 arg1 sections 1873:1880 FFPE ovarian cancer sections 1853:1880 FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI 1853:1944 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 0 108 theme Ovarian 111:117 arg1 Tissues 126:132 Ovarian Cancer Tissues 111:132 Ovarian Cancer Tissues 111:132 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 1 109 from malignancy 176:185 arg1 women 196:200 adult women 190:200 adult women 190:200 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 5 110 theme serous 1026:1031 arg1 n 1050:1050 n = 3 1050:1054 n = 3 1050:1054 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 5 110 theme serous 1026:1031 arg1 cancers 1041:1047 advanced serous ovarian cancers 1017:1047 advanced serous ovarian cancers (n = 3) 1017:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 8 111 theme graphitized 1599:1609 arg1 PGC-LC 1641:1646 PGC-LC 1641:1646 PGC-LC 1641:1646 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 8 111 theme graphitized 1599:1609 arg1 chromatography 1625:1638 porous graphitized carbon liquid chromatography 1592:1638 porous graphitized carbon liquid chromatography (PGC-LC) 1592:1647 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 6 112 from images 1241:1246 arg1 tissue 1303:1308 ovarian cancer tissue 1288:1308 ovarian cancer tissue 1288:1308 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 11 113 from abundant 2205:2212 arg1 stroma 2233:2238 the intervening stroma 2217:2238 the intervening stroma 2217:2238 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 0 114 theme Tissues 126:132 arg1 Delineation 96:106 the Delineation 92:106 the Delineation of Ovarian Cancer Tissues 92:132 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 5 115 theme cancers 1041:1047 arg1 sections 1005:1012 FFPE tissue sections 993:1012 FFPE tissue sections of advanced serous ovarian cancers (n = 3) 993:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 8 116 theme liquid 1618:1623 arg1 PGC-LC 1641:1646 PGC-LC 1641:1646 PGC-LC 1641:1646 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 8 116 theme liquid 1618:1623 arg1 chromatography 1625:1638 porous graphitized carbon liquid chromatography 1592:1638 porous graphitized carbon liquid chromatography (PGC-LC) 1592:1647 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 5 117 theme tissue 891:896 arg1 regions 898:904 tissue regions 891:904 tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) 891:972 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 8 118 theme electrospray 1671:1682 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 5 119 theme tissue 998:1003 arg1 sections 1005:1012 FFPE tissue sections 993:1012 FFPE tissue sections of advanced serous ovarian cancers (n = 3) 993:1055 In this study, tissue regions of interest (e.g. tumor, stroma, adipose tissue and necrotic areas) were isolated from FFPE tissue sections of advanced serous ovarian cancers (n = 3). 27412689 0 120 theme Mass 23:26 arg1 Spectrometry 28:39 N-glycan MALDI Imaging Mass Spectrometry 0:39 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue 0:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 3 121 dep development 511:521 arg1 the 507:509 the 507:509 the 507:509 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 6 122 theme complementary 1102:1114 arg1 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS 1058:1073 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 6 122 theme complementary 1102:1114 arg1 techniques 1116:1125 complementary techniques 1102:1125 complementary techniques 1102:1125 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 6 122 theme complementary 1102:1114 arg1 MALDI-MSI 1079:1087 MALDI-MSI 1079:1087 MALDI-MSI 1079:1087 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 4 123 theme matrix-assisted 633:647 arg1 spectrometry 690:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry 633:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 3 124 theme unearth 461:467 arg1 changes 485:491 unearth tissue-specific changes 461:491 unearth tissue-specific changes that occur in the development and progression of ovarian cancer 461:555 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 8 125 theme collision 1653:1661 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 7 126 theme ovarian 1450:1456 arg1 tissue 1465:1470 FFPE ovarian cancer tissue regions 1445:1478 FFPE ovarian cancer tissue regions 1445:1478 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 0 127 theme Paraffin-Embedded 59:75 arg1 Tissue 77:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 Formalin-Fixed Paraffin-Embedded Tissue 44:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 27412689 3 128 theme changes 449:455 arg1 investigation 412:424 spatio-temporal investigation 396:424 spatio-temporal investigation of these glycosylation changes 396:455 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 4 129 theme N-linked 607:614 arg1 glycans 616:622 tissue-specific N-linked glycans 591:622 tissue-specific N-linked glycans 591:622 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 9 130 used used 1785:1788 arg2 profiles 1738:1745 The N-glycan profiles 1725:1745 The N-glycan profiles identified by this analysis 1725:1773 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 1 131 theme fatal 155:159 arg1 cancer 143:148 Ovarian cancer 135:148 Ovarian cancer 135:148 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 1 131 theme fatal 155:159 arg1 malignancy 176:185 a fatal gynaecological malignancy 153:185 a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30% 153:251 Ovarian cancer is a fatal gynaecological malignancy in adult women with a five-year overall survival rate of only 30%. 27412689 3 132 theme cancer 550:555 arg1 progression 527:537 progression 527:537 progression 527:537 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 3 132 theme cancer 550:555 arg1 development 511:521 development 511:521 development 511:521 Therefore, spatio-temporal investigation of these glycosylation changes may unearth tissue-specific changes that occur in the development and progression of ovarian cancer. 27412689 9 133 theme N-glycan 1729:1736 arg1 profiles 1738:1745 The N-glycan profiles 1725:1745 The N-glycan profiles identified by this analysis 1725:1773 The N-glycan profiles identified by this analysis were then used to determine the location and distribution of each N-glycan on FFPE ovarian cancer sections that were treated with PNGase F using high resolution MALDI-MSI. 27412689 11 134 theme high 2076:2079 arg1 glycans 2089:2095 high mannose glycans 2076:2095 high mannose glycans 2076:2095 For example, high mannose glycans were predominantly expressed in the tumor tissue region whereas complex/hybrid N-glycans were significantly abundant in the intervening stroma. 27412689 4 135 theme mass 685:688 arg1 spectrometry 690:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry 633:701 matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) 633:715 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 10 136 theme N-glycan 1981:1988 arg1 structures 1990:1999 N-glycan structures 1981:1999 N-glycan structures 1981:1999 A tissue-specific distribution of N-glycan structures identified particular regions of the ovarian cancer sections. 27412689 6 137 from information 1158:1168 arg1 glycans 1193:1199 the tissue-specific glycans 1173:1199 the tissue-specific glycans 1173:1199 PGC-LC-ESI-MS/MS and MALDI-MSI were used as complementary techniques to firstly generate structural information on the tissue-specific glycans in order to then obtain high resolution images of the glycan structure distribution in ovarian cancer tissue. 27412689 2 138 theme pattern 346:352 arg1 alterations 354:364 extensive protein glycosylation pattern alterations 314:364 extensive protein glycosylation pattern alterations in ovarian cancer 314:382 Glycomic and glycoproteomic profiling studies have reported extensive protein glycosylation pattern alterations in ovarian cancer. 27412689 4 139 dep paraffin-embedded 735:751 arg1 FFPE 754:757 FFPE 754:757 FFPE 754:757 A novel method for investigating tissue-specific N-linked glycans is using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) on formalin-fixed paraffin-embedded (FFPE) tissue sections that can spatially profile N-glycan compositions released from proteins in tissue-specific regions. 27412689 7 140 theme glycan 1324:1329 arg1 repertoires 1331:1341 The N-linked glycan repertoires 1311:1341 The N-linked glycan repertoires carried by the proteins in these tissue regions 1311:1389 The N-linked glycan repertoires carried by the proteins in these tissue regions were structurally characterized for the first time in FFPE ovarian cancer tissue regions, using enzymatic peptide-N-glycosidase F (PNGase F) release of N-glycans. 27412689 8 141 theme fragmentation 1701:1713 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 8 142 theme MS 1698:1699 arg1 analysis 1715:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 collision induced electrospray negative mode MS fragmentation analysis 1653:1722 The released glycans were analyzed by porous graphitized carbon liquid chromatography (PGC-LC) and collision induced electrospray negative mode MS fragmentation analysis. 27412689 0 143 theme MALDI 9:13 arg1 Spectrometry 28:39 N-glycan MALDI Imaging Mass Spectrometry 0:39 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue 0:82 N-glycan MALDI Imaging Mass Spectrometry on Formalin-Fixed Paraffin-Embedded Tissue Enables the Delineation of Ovarian Cancer Tissues. 25820733 2 0 theme Such 383:386 arg1 analyses 388:395 Such analyses 383:395 Such analyses 383:395 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 2 1 theme peptide 503:509 arg1 backbone 511:518 the peptide backbone 499:518 the peptide backbone for separate analyses 499:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 1 2 theme intact 369:374 arg1 state 376:380 their intact state 363:380 their intact state 363:380 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 4 3 theme site-specific 737:749 arg1 approach 751:758 a site-specific approach 735:758 a site-specific approach for intact glycopeptide analysis 735:791 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 4 4 theme many 632:635 arg1 cases 637:641 many cases 632:641 many cases 632:641 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 5 5 theme collision-induced 1068:1084 arg1 CID 1100:1102 CID 1100:1102 CID 1100:1102 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 5 theme collision-induced 1068:1084 arg1 dissociation 1086:1097 collision-induced dissociation 1068:1097 collision-induced dissociation (CID) 1068:1103 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 4 6 theme glycopeptide 771:782 arg1 analysis 784:791 intact glycopeptide analysis 764:791 intact glycopeptide analysis 764:791 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 4 7 theme intact 764:769 arg1 analysis 784:791 intact glycopeptide analysis 764:791 intact glycopeptide analysis 764:791 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 4 8 theme bacterial 659:667 arg1 sugars 669:674 unique bacterial sugars 652:674 unique bacterial sugars 652:674 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 1 9 theme Large-scale 79:89 arg1 analysis 91:98 Large-scale analysis 79:98 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry 79:160 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 2 10 theme commercial 415:424 arg1 F 467:467 e.g. protein N-glycosidase F 440:467 e.g. protein N-glycosidase F 440:467 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 2 10 theme commercial 415:424 arg1 glycosidases 426:437 commercial glycosidases 415:437 commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses 415:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 5 11 theme zwitterionic-hydrophilic 874:897 arg1 ZIC-HILIC 934:942 ZIC-HILIC 934:942 ZIC-HILIC 934:942 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 11 theme zwitterionic-hydrophilic 874:897 arg1 chromatography 918:931 zwitterionic-hydrophilic interaction liquid chromatography 874:931 zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) 874:943 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 12 theme protein 103:109 arg1 N- 111:112 protein N- 103:112 protein N- 103:112 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 13 theme structure 1179:1187 arg1 elucidation 1189:1199 glycan structure elucidation 1172:1199 glycan structure elucidation 1172:1199 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 14 theme interaction 899:909 arg1 ZIC-HILIC 934:942 ZIC-HILIC 934:942 ZIC-HILIC 934:942 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 14 theme interaction 899:909 arg1 chromatography 918:931 zwitterionic-hydrophilic interaction liquid chromatography 874:931 zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) 874:943 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 15 theme glycosylated 302:313 arg1 peptides 315:322 their formerly glycosylated peptides 287:322 their formerly glycosylated peptides (glycoproteomics) 287:340 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 1 15 theme glycosylated 302:313 arg1 glycoproteomics 325:339 glycoproteomics 325:339 glycoproteomics 325:339 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 16 theme mass 1034:1037 arg1 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 16 theme mass 1034:1037 arg1 spectrometry 1039:1050 electrospray ionization tandem mass spectrometry 1003:1050 electrospray ionization tandem mass spectrometry (LC-MS/MS) 1003:1061 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 17 theme N- 111:112 arg1 analysis 91:98 Large-scale analysis 79:98 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry 79:160 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 18 gly glycopeptide 1205:1216 arg2 glycopeptide 1205:1216 glycopeptide 1205:1216 glycopeptide 1205:1216 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 2 19 theme N-glycosidase 453:465 arg1 F 467:467 e.g. protein N-glycosidase F 440:467 e.g. protein N-glycosidase F 440:467 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 2 19 theme N-glycosidase 453:465 arg1 glycosidases 426:437 commercial glycosidases 415:437 commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses 415:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 3 20 theme widespread 606:615 arg1 phenomenon 617:626 a widespread phenomenon 604:626 a widespread phenomenon 604:626 Bacterial glycosylation has only recently been identified as a widespread phenomenon. 25820733 3 20 theme widespread 606:615 arg1 glycosylation 553:565 Bacterial glycosylation 543:565 Bacterial glycosylation 543:565 Bacterial glycosylation has only recently been identified as a widespread phenomenon. 25820733 5 21 theme glycan 1172:1177 arg1 elucidation 1189:1199 glycan structure elucidation 1172:1199 glycan structure elucidation 1172:1199 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 2 22 theme protein 445:451 arg1 F 467:467 e.g. protein N-glycosidase F 440:467 e.g. protein N-glycosidase F 440:467 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 2 22 theme protein 445:451 arg1 glycosidases 426:437 commercial glycosidases 415:437 commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses 415:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 0 23 theme glycopeptides 43:55 arg1 identification 15:28 identification 15:28 identification 15:28 Enrichment and identification of bacterial glycopeptides by mass spectrometry. 25820733 0 23 theme glycopeptides 43:55 arg1 Enrichment 0:9 Enrichment 0:9 Enrichment 0:9 Enrichment and identification of bacterial glycopeptides by mass spectrometry. 25820733 5 24 theme ionization 1016:1025 arg1 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 24 theme ionization 1016:1025 arg1 spectrometry 1039:1050 electrospray ionization tandem mass spectrometry 1003:1050 electrospray ionization tandem mass spectrometry (LC-MS/MS) 1003:1061 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 4 25 gly glycopeptide 771:782 arg2 glycopeptide 771:782 intact glycopeptide analysis 764:791 intact glycopeptide analysis 764:791 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 4 26 theme enzymatic 685:693 arg1 removal 695:701 enzymatic removal 685:701 enzymatic removal 685:701 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 5 27 theme energy 1116:1121 arg1 HCD 1149:1151 HCD 1149:1151 HCD 1149:1151 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 27 theme energy 1116:1121 arg1 dissociation 1135:1146 higher energy collisional dissociation 1109:1146 higher energy collisional dissociation (HCD) 1109:1152 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 2 28 theme e.g. 440:443 arg1 F 467:467 e.g. protein N-glycosidase F 440:467 e.g. protein N-glycosidase F 440:467 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 2 28 theme e.g. 440:443 arg1 glycosidases 426:437 commercial glycosidases 415:437 commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses 415:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 0 29 theme bacterial 33:41 arg1 glycopeptides 43:55 bacterial glycopeptides 33:55 bacterial glycopeptides 33:55 Enrichment and identification of bacterial glycopeptides by mass spectrometry. 25820733 5 30 theme collisional 1123:1133 arg1 HCD 1149:1151 HCD 1149:1151 HCD 1149:1151 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 30 theme collisional 1123:1133 arg1 dissociation 1135:1146 higher energy collisional dissociation 1109:1146 higher energy collisional dissociation (HCD) 1109:1152 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 31 gly glycosylated 302:313 arg1 peptides 315:322 their formerly glycosylated peptides 287:322 their formerly glycosylated peptides (glycoproteomics) 287:340 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 1 31 gly glycosylated 302:313 arg1 glycoproteomics 325:339 glycoproteomics 325:339 glycoproteomics 325:339 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 0 32 theme mass 60:63 arg1 spectrometry 65:76 mass spectrometry 60:76 mass spectrometry 60:76 Enrichment and identification of bacterial glycopeptides by mass spectrometry. 25820733 5 33 theme liquid 970:975 arg1 chromatography 977:990 liquid chromatography 970:990 liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification 970:1231 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 34 theme O-linked 118:125 arg1 glycosylation 127:139 O-linked glycosylation 118:139 O-linked glycosylation 118:139 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 35 theme glycopeptides 854:866 arg1 enrichment 830:839 the enrichment 826:839 the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification 826:1231 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 36 dep dissociation 1086:1097 arg1 fragmentation 1154:1166 fragmentation 1154:1166 fragmentation 1154:1166 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 37 theme glycosylation 127:139 arg1 analysis 91:98 Large-scale analysis 79:98 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry 79:160 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 38 theme higher 1109:1114 arg1 HCD 1149:1151 HCD 1149:1151 HCD 1149:1151 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 38 theme higher 1109:1114 arg1 dissociation 1135:1146 higher energy collisional dissociation 1109:1146 higher energy collisional dissociation (HCD) 1109:1152 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 39 link O-linked 118:125 arg1 glycosylation 127:139 O-linked glycosylation 118:139 O-linked glycosylation 118:139 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 5 40 theme tandem 1027:1032 arg1 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 40 theme tandem 1027:1032 arg1 spectrometry 1039:1050 electrospray ionization tandem mass spectrometry 1003:1050 electrospray ionization tandem mass spectrometry (LC-MS/MS) 1003:1061 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 41 dep elucidation 1189:1199 arg1 identification 1218:1231 identification 1218:1231 identification 1218:1231 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 42 theme electrospray 1003:1014 arg1 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 LC-MS/MS 1053:1060 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 42 theme electrospray 1003:1014 arg1 spectrometry 1039:1050 electrospray ionization tandem mass spectrometry 1003:1050 electrospray ionization tandem mass spectrometry (LC-MS/MS) 1003:1061 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 0 43 gly glycopeptides 43:55 arg2 glycopeptides 43:55 bacterial glycopeptides 33:55 bacterial glycopeptides 33:55 Enrichment and identification of bacterial glycopeptides by mass spectrometry. 25820733 1 44 theme mass 144:147 arg1 spectrometry 149:160 mass spectrometry 144:160 mass spectrometry 144:160 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 1 45 theme glycan 253:258 arg1 structures 260:269 glycan structures 253:269 glycan structures (glycomics) 253:281 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 1 45 theme glycan 253:258 arg1 glycomics 272:280 glycomics 272:280 glycomics 272:280 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 2 46 theme separate 524:531 arg1 analyses 533:540 separate analyses 524:540 separate analyses 524:540 Such analyses depend heavily on commercial glycosidases (e.g. protein N-glycosidase F) that can remove glycans from the peptide backbone for separate analyses. 25820733 5 47 gly glycopeptides 854:866 arg2 glycopeptides 854:866 bacterial glycopeptides 844:866 bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification 844:1231 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 1 48 theme parallel 212:219 arg1 approaches 221:230 parallel approaches 212:230 parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state 212:380 Large-scale analysis of protein N- and O-linked glycosylation by mass spectrometry has traditionally been performed in eukaryotes by parallel approaches aimed at elucidating glycan structures (glycomics) and their formerly glycosylated peptides (glycoproteomics) without reference to their intact state. 25820733 3 49 theme Bacterial 543:551 arg1 phenomenon 617:626 a widespread phenomenon 604:626 a widespread phenomenon 604:626 Bacterial glycosylation has only recently been identified as a widespread phenomenon. 25820733 3 49 theme Bacterial 543:551 arg1 glycosylation 553:565 Bacterial glycosylation 543:565 Bacterial glycosylation 543:565 Bacterial glycosylation has only recently been identified as a widespread phenomenon. 25820733 4 50 theme unique 652:657 arg1 sugars 669:674 unique bacterial sugars 652:674 unique bacterial sugars 652:674 In many cases however, unique bacterial sugars preclude enzymatic removal, therefore ultimately requiring a site-specific approach for intact glycopeptide analysis. 25820733 5 51 theme bacterial 844:852 arg1 glycopeptides 854:866 bacterial glycopeptides 844:866 bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification 844:1231 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 52 theme liquid 911:916 arg1 ZIC-HILIC 934:942 ZIC-HILIC 934:942 ZIC-HILIC 934:942 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 25820733 5 52 theme liquid 911:916 arg1 chromatography 918:931 zwitterionic-hydrophilic interaction liquid chromatography 874:931 zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) 874:943 Here, we describe protocols for the enrichment of bacterial glycopeptides using zwitterionic-hydrophilic interaction liquid chromatography (ZIC-HILIC) and their analysis using liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS/MS) with collision-induced dissociation (CID) and higher energy collisional dissociation (HCD) fragmentation for glycan structure elucidation and glycopeptide identification. 27114034 6 0 theme glycan-array 944:955 arg1 binding 957:963 glycan-array binding 944:963 glycan-array binding 944:963 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 2 1 dep clades 409:414 arg1 B 419:419 B 419:419 B 419:419 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 2 1 dep clades 409:414 arg1 G 426:426 G 426:426 G 426:426 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 2 1 dep clades 409:414 arg1 A 416:416 A 416:416 A 416:416 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 2 1 dep clades 409:414 arg1 clades 409:414 clades A, B, and G 409:426 clades A, B, and G 409:426 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 1 2 link N-linked 146:153 arg1 oligosaccharides 155:170 ∼90 N-linked oligosaccharides 142:170 ∼90 N-linked oligosaccharides 142:170 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 3 3 theme most 690:693 arg1 neutralization 713:726 most antibody-mediated neutralization 690:726 most antibody-mediated neutralization 690:726 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 6 4 theme broadly 1092:1098 arg1 antibodies 1113:1122 known broadly neutralizing antibodies 1086:1122 known broadly neutralizing antibodies that target the glycan-shielded trimer 1086:1161 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 1 5 from component 224:232 arg1 immunity 262:269 humoral immunity 254:269 humoral immunity 254:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 6 6 theme glycan-shielded 1140:1154 arg1 trimer 1156:1161 the glycan-shielded trimer 1136:1161 the glycan-shielded trimer 1136:1161 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 6 7 theme oligosaccharide 1011:1025 arg1 affinity 1027:1034 oligosaccharide affinity 1011:1034 oligosaccharide affinity 1011:1034 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 5 8 theme Crowded 812:818 arg1 glycans 834:840 Crowded and dispersed glycans 812:840 Crowded and dispersed glycans 812:840 Crowded and dispersed glycans are differently ordered, conserved, processed, and recognized by antibody. 27114034 2 9 theme glycosylated 379:390 arg1 trimers 396:402 fully glycosylated Env trimers 373:402 fully glycosylated Env trimers 373:402 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 3 10 theme protein 639:645 arg1 component 647:655 the protein component 635:655 the protein component of Env 635:662 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 4 11 theme interactions 798:809 arg1 taxonomy 763:770 a taxonomy 761:770 a taxonomy of N-linked glycan-glycan interactions 761:809 The revealed features delineate a taxonomy of N-linked glycan-glycan interactions. 27114034 3 12 dep HIV-1-glycan 505:516 arg1 shield 518:523 shield 518:523 shield 518:523 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 0 13 theme Trimeric 0:7 arg1 Structures 19:28 Trimeric HIV-1-Env Structures 0:28 Trimeric HIV-1-Env Structures 0:28 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 4 14 theme glycan-glycan 784:796 arg1 interactions 798:809 N-linked glycan-glycan interactions 775:809 N-linked glycan-glycan interactions 775:809 The revealed features delineate a taxonomy of N-linked glycan-glycan interactions. 27114034 6 15 from diversity 998:1006 arg1 affinity 1027:1034 oligosaccharide affinity 1011:1034 oligosaccharide affinity 1011:1034 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 4 16 theme N-linked 775:782 arg1 interactions 798:809 N-linked glycan-glycan interactions 775:809 N-linked glycan-glycan interactions 775:809 The revealed features delineate a taxonomy of N-linked glycan-glycan interactions. 27114034 5 17 theme dispersed 824:832 arg1 glycans 834:840 Crowded and dispersed glycans 812:840 Crowded and dispersed glycans 812:840 Crowded and dispersed glycans are differently ordered, conserved, processed, and recognized by antibody. 27114034 1 18 theme key 220:222 arg1 component 224:232 a key component 218:232 a key component of HIV evasion from humoral immunity 218:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 2 19 theme 3.4-3.7 Å 455:463 arg1 resolution 465:474 3.4-3.7 Å resolution 455:474 3.4-3.7 Å resolution 455:474 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 2 20 dep glycan 342:347 arg1 shield 349:354 shield 349:354 shield 349:354 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 3 21 theme Env 660:662 arg1 component 647:655 the protein component 635:655 the protein component of Env 635:662 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 3 22 theme oligosaccharides 563:578 arg1 network 539:545 a network 537:545 a network 537:545 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 6 23 theme neutralizing 1100:1111 arg1 antibodies 1113:1122 known broadly neutralizing antibodies 1086:1122 known broadly neutralizing antibodies that target the glycan-shielded trimer 1086:1161 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 1 24 theme N-linked 146:153 arg1 oligosaccharides 155:170 ∼90 N-linked oligosaccharides 142:170 ∼90 N-linked oligosaccharides 142:170 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 1 25 dep half 197:200 arg1 mass 206:209 its mass 202:209 half its mass 197:209 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 0 26 dep Clades 57:62 arg1 B 67:67 B 67:67 B 67:67 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 0 26 dep Clades 57:62 arg1 G 74:74 G 74:74 G 74:74 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 0 26 dep Clades 57:62 arg1 A 64:64 A 64:64 A 64:64 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 0 26 dep Clades 57:62 arg1 Clades 57:62 Clades A, B, and G 57:74 Clades A, B, and G 57:74 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 1 27 theme oligosaccharides 155:170 arg1 glycan 125:130 a glycan shield 123:137 a glycan shield 123:137 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 0 28 theme Glycan 37:42 arg1 Shields 44:50 Glycan Shields 37:50 Glycan Shields 37:50 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 1 29 theme HIV 237:239 arg1 evasion 241:247 HIV evasion 237:247 HIV evasion from humoral immunity 237:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 1 30 from immunity 262:269 arg1 component 224:232 a key component 218:232 a key component of HIV evasion from humoral immunity 218:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 1 30 from immunity 262:269 arg1 evasion 241:247 HIV evasion 237:247 HIV evasion from humoral immunity 237:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 1 31 theme evasion 241:247 arg1 component 224:232 a key component 218:232 a key component of HIV evasion from humoral immunity 218:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 3 32 theme interlocking 550:561 arg1 oligosaccharides 563:578 interlocking oligosaccharides 550:578 interlocking oligosaccharides 550:578 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 1 33 theme HIV-1-envelope 81:94 arg1 trimer 102:107 The HIV-1-envelope (Env) trimer 77:107 The HIV-1-envelope (Env) trimer 77:107 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 2 34 theme Env 392:394 arg1 trimers 396:402 fully glycosylated Env trimers 373:402 fully glycosylated Env trimers 373:402 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 1 35 dep glycan 125:130 arg1 shield 132:137 shield 132:137 shield 132:137 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 6 36 theme known 1086:1090 arg1 antibodies 1113:1122 known broadly neutralizing antibodies 1086:1122 known broadly neutralizing antibodies that target the glycan-shielded trimer 1086:1161 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 1 37 theme humoral 254:260 arg1 immunity 262:269 humoral immunity 254:269 humoral immunity 254:269 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 3 38 theme glycan 606:611 arg1 crowding 613:620 glycan crowding 606:620 glycan crowding 606:620 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 27114034 1 39 theme Env 97:99 arg1 trimer 102:107 The HIV-1-envelope (Env) trimer 77:107 The HIV-1-envelope (Env) trimer 77:107 The HIV-1-envelope (Env) trimer is covered by a glycan shield of ∼90 N-linked oligosaccharides, which comprises roughly half its mass and is a key component of HIV evasion from humoral immunity. 27114034 2 40 gly glycosylated 379:390 arg1 trimers 396:402 fully glycosylated Env trimers 373:402 fully glycosylated Env trimers 373:402 To understand how antibodies can overcome the barriers imposed by the glycan shield, we crystallized fully glycosylated Env trimers from clades A, B, and G, visualizing the shield at 3.4-3.7 Å resolution. 27114034 6 41 from requirement 1042:1052 arg1 affinity 1027:1034 oligosaccharide affinity 1011:1034 oligosaccharide affinity 1011:1034 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 6 42 theme molecular 969:977 arg1 dynamics 979:986 molecular dynamics 969:986 molecular dynamics 969:986 The structures, along with glycan-array binding and molecular dynamics, reveal a diversity in oligosaccharide affinity and a requirement for accommodating glycans among known broadly neutralizing antibodies that target the glycan-shielded trimer. 27114034 4 43 theme revealed 733:740 arg1 features 742:749 The revealed features 729:749 The revealed features 729:749 The revealed features delineate a taxonomy of N-linked glycan-glycan interactions. 27114034 4 44 link N-linked 775:782 arg1 interactions 798:809 N-linked glycan-glycan interactions 775:809 N-linked glycan-glycan interactions 775:809 The revealed features delineate a taxonomy of N-linked glycan-glycan interactions. 27114034 0 45 theme HIV-1-Env 9:17 arg1 Structures 19:28 Trimeric HIV-1-Env Structures 0:28 Trimeric HIV-1-Env Structures 0:28 Trimeric HIV-1-Env Structures Define Glycan Shields from Clades A, B, and G. 27114034 3 46 theme antibody-mediated 695:711 arg1 neutralization 713:726 most antibody-mediated neutralization 690:726 most antibody-mediated neutralization 690:726 These structures reveal the HIV-1-glycan shield to comprise a network of interlocking oligosaccharides, substantially ordered by glycan crowding, that encase the protein component of Env and enable HIV-1 to avoid most antibody-mediated neutralization. 24326091 7 0 theme end 1525:1527 arg1 integrity 1499:1507 the structural integrity 1484:1507 the structural integrity of the reducing end of the oligosaccharide 1484:1550 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 9 1 used utilized 1896:1903 arg2 example 1763:1769 an example 1760:1769 an example of a biochemical study using functionalized high mannose glycosylated asparagine 1760:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 1 used utilized 1896:1903 arg2 assay 1881:1885 a fluorescence polarization assay 1853:1885 a fluorescence polarization assay 1853:1885 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 2 2 theme protected 360:368 arg1 residue 384:390 an Fmoc protected aspartic acid residue 352:390 an Fmoc protected aspartic acid residue 352:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 6 3 theme 4-pentynoic 1210:1220 arg1 alkyne 1202:1207 an alkyne 1199:1207 an alkyne (4-pentynoic acid) 1199:1226 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 6 3 theme 4-pentynoic 1210:1220 arg1 acid 1222:1225 4-pentynoic acid 1210:1225 4-pentynoic acid 1210:1225 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 8 4 theme native 1665:1670 arg1 presentation 1672:1683 the native presentation 1661:1683 the native presentation of N-linked glycan which may aid in biochemical and structural studies 1661:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 4 theme native 1665:1670 arg1 linker 1596:1601 a linker 1594:1601 a linker between the glycan and the functional group 1594:1645 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 4 theme native 1665:1670 arg1 chain 1573:1577 The asparagine side chain 1553:1577 The asparagine side chain 1553:1577 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 6 5 theme N-terminal 1291:1300 arg1 amine 1302:1306 the N-terminal amine 1287:1306 the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group 1287:1376 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 3 6 theme protecting 504:513 arg1 group 515:519 a 2-phenylisopropyl protecting group 484:519 a 2-phenylisopropyl protecting group 484:519 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 7 7 gly glycosylated 1442:1453 arg1 asparagine 1455:1464 the glycosylated asparagine 1438:1464 the glycosylated asparagine 1438:1464 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 5 8 theme glycosylation 938:950 arg1 yield 952:956 excellent glycosylation yield 928:956 excellent glycosylation yield 928:956 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 4 9 theme glycosylated 851:862 arg1 asparagine 864:873 N-linked high mannose glycosylated asparagine 829:873 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 9 10 theme Concanavalin 1940:1951 arg1 A 1953:1953 the lectin Concanavalin A 1929:1953 the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1929:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 10 theme Concanavalin 1940:1951 arg1 ConA 1956:1959 ConA 1956:1959 ConA 1956:1959 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 3 11 theme selective 535:543 arg1 deprotection 545:556 selective deprotection 535:556 selective deprotection 535:556 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 3 12 theme glycosylation 582:594 arg1 introduction 566:577 the introduction 562:577 the introduction of glycosylation 562:594 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 7 13 dep functional 1399:1408 arg1 handles 1410:1416 handles 1410:1416 handles 1410:1416 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 6 14 theme Fmoc 1356:1359 arg1 group 1372:1376 the Fmoc protecting group 1352:1376 the Fmoc protecting group 1352:1376 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 4 15 gly glycosylated 851:862 arg1 asparagine 864:873 N-linked high mannose glycosylated asparagine 829:873 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 1 16 dep functional 264:273 arg1 handles 275:281 handles 275:281 handles 275:281 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 9 17 theme 5-carboxyfluorescein 1968:1987 arg1 mannose 2002:2008 5-carboxyfluorescein labeled high mannose 1968:2008 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 18 theme N-glycosidic 664:675 arg1 linkage 677:683 an N-glycosidic linkage 661:683 an N-glycosidic linkage 661:683 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 5 19 theme coupling 1111:1118 arg1 reaction 1120:1127 the on-resin coupling reaction 1098:1127 the on-resin coupling reaction 1098:1127 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 9 20 theme high 1997:2000 arg1 mannose 2002:2008 5-carboxyfluorescein labeled high mannose 1968:2008 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 5 21 theme excess 1055:1060 arg1 oligosaccharides 1062:1077 excess oligosaccharides 1055:1077 excess oligosaccharides 1055:1077 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 9 22 theme glycosylated 2010:2021 arg1 asparagine 2023:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 23 theme resin 738:742 arg1 chain 725:729 the free side chain 711:729 the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine 711:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 6 24 theme on-resin 1322:1329 arg1 amine 1302:1306 the N-terminal amine 1287:1306 the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group 1287:1376 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 1 25 theme solid 132:136 arg1 synthesis 144:152 solid phase synthesis 132:152 solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 132:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 5 26 theme coupling 910:917 arg1 use 880:882 The use 876:882 The use of on-resin glycosylamine coupling 876:917 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 9 27 theme biochemical 1776:1786 arg1 study 1788:1792 a biochemical study 1774:1792 a biochemical study using functionalized high mannose glycosylated asparagine 1774:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 28 theme side 720:723 arg1 chain 725:729 the free side chain 711:729 the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine 711:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 8 29 link N-linked 1688:1695 arg1 glycan 1697:1702 N-linked glycan 1688:1702 N-linked glycan 1688:1702 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 1 30 theme N-linked 157:164 arg1 glycoconjugates 166:180 N-linked glycoconjugates 157:180 N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 157:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 4 31 theme large 771:775 arg1 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 4 31 theme large 771:775 arg1 oligosaccharide 790:804 a large high mannose oligosaccharide 769:804 a large high mannose oligosaccharide 769:804 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 9 32 theme high 1815:1818 arg1 mannose 1820:1826 functionalized high mannose 1800:1826 functionalized high mannose glycosylated asparagine 1800:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 33 theme mannose 782:788 arg1 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 4 33 theme mannose 782:788 arg1 oligosaccharide 790:804 a large high mannose oligosaccharide 769:804 a large high mannose oligosaccharide 769:804 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 3 34 theme side 440:443 arg1 chain 445:449 The side chain 436:449 The side chain of aspartic acid 436:466 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 6 35 theme group 1372:1376 arg1 removal 1341:1347 the removal 1337:1347 the removal of the Fmoc protecting group 1337:1376 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 5 36 theme excellent 928:936 arg1 yield 952:956 excellent glycosylation yield 928:956 excellent glycosylation yield 928:956 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 5 37 theme oligosaccharides 999:1014 arg1 types 990:994 other types 984:994 other types of oligosaccharides 984:1014 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 4 38 link N-linked 829:836 arg1 mannose 843:849 N-linked high mannose 829:849 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 9 39 theme glycosylated 1828:1839 arg1 asparagine 1841:1850 functionalized high mannose glycosylated asparagine 1800:1850 functionalized high mannose glycosylated asparagine 1800:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 8 40 theme side 1568:1571 arg1 presentation 1672:1683 the native presentation 1661:1683 the native presentation of N-linked glycan which may aid in biochemical and structural studies 1661:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 40 theme side 1568:1571 arg1 linker 1596:1601 a linker 1594:1601 a linker between the glycan and the functional group 1594:1645 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 40 theme side 1568:1571 arg1 chain 1573:1577 The asparagine side chain 1553:1577 The asparagine side chain 1553:1577 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 4 41 theme aspartic 750:757 arg1 acid 759:762 aspartic acid 750:762 aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine 750:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 0 42 theme Versatile 0:8 arg1 synthesis 19:27 Versatile on-resin synthesis 0:27 Versatile on-resin synthesis of high mannose 0:43 Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles. 24326091 3 43 theme acid 463:466 arg1 chain 445:449 The side chain 436:449 The side chain of aspartic acid 436:466 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 1 44 theme phase 138:142 arg1 synthesis 144:152 solid phase synthesis 132:152 solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 132:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 9 45 theme fluorescence 1855:1866 arg1 assay 1881:1885 a fluorescence polarization assay 1853:1885 a fluorescence polarization assay 1853:1885 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 45 theme fluorescence 1855:1866 arg1 example 1763:1769 an example 1760:1769 an example of a biochemical study using functionalized high mannose glycosylated asparagine 1760:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 2 46 theme solid 415:419 arg1 support 427:433 a solid phase support 413:433 a solid phase support 413:433 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 0 47 theme high 32:35 arg1 mannose 37:43 high mannose 32:43 high mannose 32:43 Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles. 24326091 4 48 theme high 838:841 arg1 mannose 843:849 N-linked high mannose 829:849 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 2 49 theme Fmoc 355:358 arg1 residue 384:390 an Fmoc protected aspartic acid residue 352:390 an Fmoc protected aspartic acid residue 352:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 2 50 attach attached 401:408 arg2 C-terminus 338:347 the C-terminus 334:347 the C-terminus of an Fmoc protected aspartic acid residue 334:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 2 50 attach attached 401:408 arg3 strategy 324:331 this strategy 319:331 this strategy 319:331 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 2 50 attach attached 401:408 arg1 support 427:433 a solid phase support 413:433 a solid phase support 413:433 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 4 51 theme convergent 608:617 arg1 strategy 651:658 a convergent on-resin glycosylamine coupling strategy 606:658 a convergent on-resin glycosylamine coupling strategy 606:658 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 2 52 theme residue 384:390 arg1 C-terminus 338:347 the C-terminus 334:347 the C-terminus of an Fmoc protected aspartic acid residue 334:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 8 53 theme N-linked 1688:1695 arg1 glycan 1697:1702 N-linked glycan 1688:1702 N-linked glycan 1688:1702 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 7 54 theme reducing 1516:1523 arg1 end 1525:1527 the reducing end 1512:1527 the reducing end of the oligosaccharide 1512:1550 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 8 55 theme functional 1630:1639 arg1 group 1641:1645 the functional group 1626:1645 the functional group 1626:1645 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 6 56 theme asparagine 1311:1320 arg1 on-resin 1322:1329 asparagine on-resin 1311:1329 asparagine on-resin 1311:1329 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 2 57 theme aspartic 370:377 arg1 residue 384:390 an Fmoc protected aspartic acid residue 352:390 an Fmoc protected aspartic acid residue 352:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 7 58 theme structural 1488:1497 arg1 integrity 1499:1507 the structural integrity 1484:1507 the structural integrity of the reducing end of the oligosaccharide 1484:1550 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 9 59 theme lectin 1933:1938 arg1 A 1953:1953 the lectin Concanavalin A 1929:1953 the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1929:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 59 theme lectin 1933:1938 arg1 ConA 1956:1959 ConA 1956:1959 ConA 1956:1959 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 8 60 theme biochemical 1721:1731 arg1 studies 1748:1754 biochemical and structural studies 1721:1754 biochemical and structural studies 1721:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 1 61 theme glycoconjugates 166:180 arg1 synthesis 144:152 solid phase synthesis 132:152 solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 132:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 4 62 theme mannose 843:849 arg1 asparagine 864:873 N-linked high mannose glycosylated asparagine 829:873 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 7 63 theme glycosylated 1442:1453 arg1 asparagine 1455:1464 the glycosylated asparagine 1438:1464 the glycosylated asparagine 1438:1464 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 9 64 theme A 1953:1953 arg1 binding 1918:1924 the binding 1914:1924 the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1914:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 8 65 theme structural 1737:1746 arg1 studies 1748:1754 biochemical and structural studies 1721:1754 biochemical and structural studies 1721:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 1 66 theme high 193:196 arg1 oligosaccharides 206:221 high mannose oligosaccharides 193:221 high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 193:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 7 67 theme oligosaccharide 1536:1550 arg1 end 1525:1527 the reducing end 1512:1527 the reducing end of the oligosaccharide 1512:1550 In this way, useful functional handles are introduced onto the glycosylated asparagine while maintaining the structural integrity of the reducing end of the oligosaccharide. 24326091 6 68 theme protecting 1361:1370 arg1 group 1372:1376 the Fmoc protecting group 1352:1376 the Fmoc protecting group 1352:1376 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 4 69 theme N-linked 829:836 arg1 mannose 843:849 N-linked high mannose 829:849 N-linked high mannose glycosylated asparagine 829:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 6 70 theme p-vinylbenzoic 1177:1190 arg1 alkene 1169:1174 an alkene 1166:1174 an alkene (p-vinylbenzoic acid) 1166:1196 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 6 70 theme p-vinylbenzoic 1177:1190 arg1 acid 1192:1195 p-vinylbenzoic acid 1177:1195 p-vinylbenzoic acid 1177:1195 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 5 71 theme on-resin 1102:1109 arg1 reaction 1120:1127 the on-resin coupling reaction 1098:1127 the on-resin coupling reaction 1098:1127 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 1 72 theme mannose 198:204 arg1 oligosaccharides 206:221 high mannose oligosaccharides 193:221 high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 193:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 9 73 theme labeled 1989:1995 arg1 mannose 2002:2008 5-carboxyfluorescein labeled high mannose 1968:2008 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 74 with acid 759:762 arg1 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 4 74 with acid 759:762 arg1 oligosaccharide 790:804 a large high mannose oligosaccharide 769:804 a large high mannose oligosaccharide 769:804 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 1 75 theme synthetic 112:120 arg1 route 122:126 a synthetic route 110:126 a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 110:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 9 76 theme mannose 2002:2008 arg1 asparagine 2023:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 1 77 from incorporation 240:252 arg1 N-terminus 290:299 the N-terminus 286:299 the N-terminus of asparagine 286:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 5 78 theme on-resin 887:894 arg1 coupling 910:917 on-resin glycosylamine coupling 887:917 on-resin glycosylamine coupling 887:917 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 5 79 theme glycosylamine 896:908 arg1 coupling 910:917 on-resin glycosylamine coupling 887:917 on-resin glycosylamine coupling 887:917 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 9 80 gly glycosylated 2010:2021 arg1 asparagine 2023:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 5-carboxyfluorescein labeled high mannose glycosylated asparagine 1968:2032 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 81 theme study 1788:1792 arg1 assay 1881:1885 a fluorescence polarization assay 1853:1885 a fluorescence polarization assay 1853:1885 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 81 theme study 1788:1792 arg1 example 1763:1769 an example 1760:1769 an example of a biochemical study using functionalized high mannose glycosylated asparagine 1760:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 82 theme free 715:718 arg1 chain 725:729 the free side chain 711:729 the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine 711:873 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 9 83 gly glycosylated 1828:1839 arg1 asparagine 1841:1850 functionalized high mannose glycosylated asparagine 1800:1850 functionalized high mannose glycosylated asparagine 1800:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 5 84 theme other 984:988 arg1 types 990:994 other types 984:994 other types of oligosaccharides 984:1014 The use of on-resin glycosylamine coupling provides excellent glycosylation yield, can be applied to couple other types of oligosaccharides, and also makes it possible to recover excess oligosaccharides conveniently after the on-resin coupling reaction. 24326091 9 85 theme functionalized 1800:1813 arg1 mannose 1820:1826 functionalized high mannose 1800:1826 functionalized high mannose glycosylated asparagine 1800:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 8 86 theme asparagine 1557:1566 arg1 presentation 1672:1683 the native presentation 1661:1683 the native presentation of N-linked glycan which may aid in biochemical and structural studies 1661:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 86 theme asparagine 1557:1566 arg1 linker 1596:1601 a linker 1594:1601 a linker between the glycan and the functional group 1594:1645 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 86 theme asparagine 1557:1566 arg1 chain 1573:1577 The asparagine side chain 1553:1577 The asparagine side chain 1553:1577 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 9 87 theme mannose 1820:1826 arg1 asparagine 1841:1850 functionalized high mannose glycosylated asparagine 1800:1850 functionalized high mannose glycosylated asparagine 1800:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 4 88 theme high 777:780 arg1 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 Man8GlcNAc2 807:817 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 4 88 theme high 777:780 arg1 oligosaccharide 790:804 a large high mannose oligosaccharide 769:804 a large high mannose oligosaccharide 769:804 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 0 89 theme on-resin 10:17 arg1 synthesis 19:27 Versatile on-resin synthesis 0:27 Versatile on-resin synthesis of high mannose 0:43 Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles. 24326091 1 90 theme asparagine 304:313 arg1 N-terminus 290:299 the N-terminus 286:299 the N-terminus of asparagine 286:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 0 91 dep functional 74:83 arg1 handles 85:91 handles 85:91 handles 85:91 Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles. 24326091 1 92 link N-linked 157:164 arg1 glycoconjugates 166:180 N-linked glycoconjugates 157:180 N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 157:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 2 93 theme phase 421:425 arg1 support 427:433 a solid phase support 413:433 a solid phase support 413:433 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 0 94 theme mannose 37:43 arg1 synthesis 19:27 Versatile on-resin synthesis 0:27 Versatile on-resin synthesis of high mannose 0:43 Versatile on-resin synthesis of high mannose glycosylated asparagine with functional handles. 24326091 9 95 theme polarization 1868:1879 arg1 assay 1881:1885 a fluorescence polarization assay 1853:1885 a fluorescence polarization assay 1853:1885 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 9 95 theme polarization 1868:1879 arg1 example 1763:1769 an example 1760:1769 an example of a biochemical study using functionalized high mannose glycosylated asparagine 1760:1850 As an example of a biochemical study using functionalized high mannose glycosylated asparagine, a fluorescence polarization assay has been utilized to study the binding of the lectin Concanavalin A (ConA) using 5-carboxyfluorescein labeled high mannose glycosylated asparagine. 24326091 1 96 contain containing 182:191 arg1 glycoconjugates 166:180 N-linked glycoconjugates 157:180 N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 157:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 1 96 contain containing 182:191 arg2 oligosaccharides 206:221 high mannose oligosaccharides 193:221 high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine 193:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24326091 3 97 theme aspartic 454:461 arg1 acid 463:466 aspartic acid 454:466 aspartic acid 454:466 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 6 98 dep functional 1137:1146 arg1 handles 1148:1154 handles 1148:1154 handles 1148:1154 Useful functional handles including an alkene (p-vinylbenzoic acid), an alkyne (4-pentynoic acid), biotin, and 5-carboxyfluorescein are then conjugated onto the N-terminal amine of asparagine on-resin after the removal of the Fmoc protecting group. 24326091 3 99 theme 2-phenylisopropyl 486:502 arg1 group 515:519 a 2-phenylisopropyl protecting group 484:519 a 2-phenylisopropyl protecting group 484:519 The side chain of aspartic acid is protected by a 2-phenylisopropyl protecting group, which allows selective deprotection for the introduction of glycosylation. 24326091 4 100 theme on-resin 619:626 arg1 strategy 651:658 a convergent on-resin glycosylamine coupling strategy 606:658 a convergent on-resin glycosylamine coupling strategy 606:658 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 8 101 theme glycan 1697:1702 arg1 presentation 1672:1683 the native presentation 1661:1683 the native presentation of N-linked glycan which may aid in biochemical and structural studies 1661:1754 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 101 theme glycan 1697:1702 arg1 linker 1596:1601 a linker 1594:1601 a linker between the glycan and the functional group 1594:1645 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 8 101 theme glycan 1697:1702 arg1 chain 1573:1577 The asparagine side chain 1553:1577 The asparagine side chain 1553:1577 The asparagine side chain also serves as a linker between the glycan and the functional group and preserves the native presentation of N-linked glycan which may aid in biochemical and structural studies. 24326091 4 102 theme coupling 642:649 arg1 strategy 651:658 a convergent on-resin glycosylamine coupling strategy 606:658 a convergent on-resin glycosylamine coupling strategy 606:658 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 4 103 theme glycosylamine 628:640 arg1 strategy 651:658 a convergent on-resin glycosylamine coupling strategy 606:658 a convergent on-resin glycosylamine coupling strategy 606:658 By using a convergent on-resin glycosylamine coupling strategy, an N-glycosidic linkage is successfully formed on the free side chain of the resin bound aspartic acid with a large high mannose oligosaccharide, Man8GlcNAc2, to yield N-linked high mannose glycosylated asparagine. 24326091 2 104 theme acid 379:382 arg1 residue 384:390 an Fmoc protected aspartic acid residue 352:390 an Fmoc protected aspartic acid residue 352:390 In this strategy, the C-terminus of an Fmoc protected aspartic acid residue is first attached to a solid phase support. 24326091 1 105 theme functional 264:273 arg1 incorporation 240:252 the incorporation 236:252 the incorporation of useful functional handles on the N-terminus of asparagine 236:313 Here we present a synthetic route for solid phase synthesis of N-linked glycoconjugates containing high mannose oligosaccharides which allows the incorporation of useful functional handles on the N-terminus of asparagine. 24664808 4 0 theme fragmentation 678:690 arg1 behavior 692:699 the fragmentation behavior 674:699 the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions 674:803 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 10 1 theme glycan 1739:1744 arg1 moiety 1746:1751 intact glycan moiety 1732:1751 intact glycan moiety 1732:1751 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 8 2 dep [peptide 1447:1454 arg1 e.g. 1441:1444 e.g. 1441:1444 e.g. 1441:1444 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 6 3 theme 2,4 1216:1218 arg1 ions 1244:1247 E ions 1242:1247 E ions 1242:1247 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 3 theme 2,4 1216:1218 arg1 D 1235:1235 D 1235:1235 D 1235:1235 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 3 theme 2,4 1216:1218 arg1 AR-1 1229:1232 AR-1 1229:1232 AR-1 1229:1232 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 3 theme 2,4 1216:1218 arg1 AR 1220:1221 (2,4)AR 1215:1221 (2,4)AR 1215:1221 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 1 4 theme common 199:204 arg1 approach 206:213 a common approach 197:213 a common approach for elucidating site-specific N-glycosylation 197:259 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 9 5 theme peptide 1556:1562 arg1 sequence 1564:1571 the peptide sequence 1552:1571 the peptide sequence 1552:1571 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 5 6 with types 945:949 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 11 7 theme fragmentation 1872:1884 arg1 rules 1886:1890 existing fragmentation rules 1863:1890 existing fragmentation rules suggested for N-glycans and peptides 1863:1927 Observed fragments are reasonably explained by a combination of existing fragmentation rules suggested for N-glycans and peptides. 24664808 0 8 link N-linked 46:53 arg1 glycopeptides 55:67 deprotonated N-linked glycopeptides 33:67 deprotonated N-linked glycopeptides 33:67 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 4 9 theme collision-induced 757:773 arg1 dissociation 775:786 low-energy collision-induced dissociation 746:786 low-energy collision-induced dissociation (CID) conditions 746:803 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 9 theme collision-induced 757:773 arg1 CID 789:791 CID 789:791 CID 789:791 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 5 10 with products 1038:1045 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 1 11 theme site-specific 231:243 arg1 N-glycosylation 245:259 site-specific N-glycosylation 231:259 site-specific N-glycosylation 231:259 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 6 12 dep AR 1220:1221 arg1 e.g. 1209:1212 e.g. 1209:1212 e.g. 1209:1212 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 5 13 theme glycan 956:961 arg1 ions 972:975 (i) glycan fragment ions 952:975 (i) glycan fragment ions 952:975 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 3 14 theme glycopeptides 359:371 arg1 fragmentation 331:343 fragmentation 331:343 fragmentation of protonated glycopeptides 331:371 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 5 15 theme fragment 810:817 arg1 species 823:829 The fragment ion species 806:829 The fragment ion species 806:829 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 8 16 dep [peptide-NH3-H 1476:1489 arg1 - 1492:1492 - 1492:1492 - 1492:1492 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 10 17 theme glycosylated 1634:1645 arg1 located 1654:1660 located 1654:1660 located 1654:1660 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 17 theme glycosylated 1634:1645 arg1 Asn 1647:1649 a glycosylated Asn 1632:1649 a glycosylated Asn 1632:1649 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 2 18 theme positive-ion 301:312 arg1 mode 314:317 positive-ion mode 301:317 positive-ion mode 301:317 The analysis is generally performed in positive-ion mode. 24664808 1 19 theme tandem 160:165 arg1 spectrometry 172:183 tandem mass spectrometry 160:183 tandem mass spectrometry 160:183 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 5 20 theme amino 890:894 arg1 sequence 901:908 their amino acid sequence 884:908 their amino acid sequence 884:908 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 4 21 theme large 568:572 arg1 sets 574:577 large sets 568:577 large sets of glycopeptides derived from well-characterized glycoproteins 568:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 21 theme large 568:572 arg1 glycopeptides 582:594 glycopeptides 582:594 glycopeptides derived from well-characterized glycoproteins 582:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 9 22 theme Asp 1608:1610 arg1 presence 1596:1603 the presence 1592:1603 the presence of Asp, Asn, and Glu 1592:1624 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 0 23 theme acid 90:93 arg1 composition 95:105 amino acid composition 84:105 amino acid composition 84:105 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 5 24 theme fragment 995:1002 arg1 ions 1004:1007 glycan-lost fragment ions 983:1007 glycan-lost fragment ions 983:1007 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 3 25 theme few 417:419 arg1 studies 421:427 few studies 417:427 few studies 417:427 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 3 26 gly glycopeptides 459:471 arg2 glycopeptides 459:471 deprotonated glycopeptides 446:471 deprotonated glycopeptides 446:471 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 4 27 gly glycopeptides 726:738 arg2 glycopeptides 726:738 deprotonated N-linked glycopeptides 704:738 deprotonated N-linked glycopeptides 704:738 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 5 28 theme secondary 1019:1027 arg1 products 1038:1045 their secondary cleavage products 1013:1045 their secondary cleavage products 1013:1045 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 5 29 dep ions 1004:1007 arg1 ii 979:980 ii 979:980 ii 979:980 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 3 30 theme negative-ion 500:511 arg1 mode 513:516 negative-ion mode 500:516 negative-ion mode analysis 500:525 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 5 31 theme fragment 1058:1065 arg1 ions 1067:1070 fragment ions 1058:1070 (iii) fragment ions with intact glycan moiety 1052:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 6 32 theme peptide 1147:1153 arg1 sequence 1155:1162 a short peptide sequence 1139:1162 a short peptide sequence 1139:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 33 contain having 1132:1137 arg1 glycopeptides 1118:1130 glycopeptides 1118:1130 glycopeptides having a short peptide sequence 1118:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 33 contain having 1132:1137 arg2 sequence 1155:1162 a short peptide sequence 1139:1162 a short peptide sequence 1139:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 5 34 with ions 1004:1007 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 0 35 theme Fragmentation 0:12 arg1 characteristics 14:28 Fragmentation characteristics 0:28 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 0:119 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 7 36 theme glycan 1310:1315 arg1 moiety 1317:1322 the glycan moiety 1306:1322 the glycan moiety 1306:1322 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 6 37 gly glycopeptides 1118:1130 arg2 glycopeptides 1118:1130 glycopeptides 1118:1130 glycopeptides having a short peptide sequence 1118:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 3 38 gly glycopeptide 540:551 arg2 glycopeptide 540:551 glycopeptide signals 540:559 glycopeptide signals 540:559 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 1 39 theme Glycopeptide 121:132 arg1 analysis 145:152 Glycopeptide structural analysis 121:152 Glycopeptide structural analysis using tandem mass spectrometry 121:183 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 6 40 theme glycopeptides 1118:1130 arg1 spectra 1107:1113 The CID spectra 1099:1113 The CID spectra of glycopeptides having a short peptide sequence 1099:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 8 41 theme peptide 1381:1387 arg1 sequences 1389:1397 long peptide sequences 1376:1397 long peptide sequences 1376:1397 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 3 42 theme analysis 518:525 arg1 usefulness 486:495 the usefulness 482:495 the usefulness of negative-ion mode analysis in detecting glycopeptide signals 482:559 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 0 43 theme N-linked 46:53 arg1 glycopeptides 55:67 deprotonated N-linked glycopeptides 33:67 deprotonated N-linked glycopeptides 33:67 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 8 44 gly glycopeptides 1347:1359 arg2 glycopeptides 1347:1359 glycopeptides 1347:1359 glycopeptides with medium or long peptide sequences 1347:1397 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 10 45 located located 1654:1660 arg2 located 1654:1660 located 1654:1660 located 1654:1660 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 45 located located 1654:1660 arg1 C-terminus 1669:1678 the C-terminus 1665:1678 the C-terminus 1665:1678 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 45 located located 1654:1660 arg2 Asn 1647:1649 a glycosylated Asn 1632:1649 a glycosylated Asn 1632:1649 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 4 46 link N-linked 717:724 arg1 glycopeptides 726:738 deprotonated N-linked glycopeptides 704:738 deprotonated N-linked glycopeptides 704:738 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 7 47 theme structural 1283:1292 arg1 features 1294:1301 detailed structural features 1274:1301 detailed structural features of the glycan moiety such as branching 1274:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 7 47 theme structural 1283:1292 arg1 branching 1332:1340 branching 1332:1340 branching 1332:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 4 48 theme well-characterized 609:626 arg1 glycoproteins 628:640 well-characterized glycoproteins 609:640 well-characterized glycoproteins 609:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 8 49 theme major 1404:1408 arg1 type 1425:1428 type (ii) ions (e.g., [peptide + (0,2)X0-H](-) 1425:1470 type 1425:1428 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 8 49 theme major 1404:1408 arg1 fragments 1410:1418 the major fragments 1400:1418 the major fragments 1400:1418 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 0 50 gly glycopeptides 55:67 arg2 glycopeptides 55:67 deprotonated N-linked glycopeptides 33:67 deprotonated N-linked glycopeptides 33:67 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 3 51 gly glycopeptides 359:371 arg2 glycopeptides 359:371 protonated glycopeptides 348:371 protonated glycopeptides 348:371 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 5 52 theme glycan 1084:1089 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 4 53 theme glycopeptides 726:738 arg1 behavior 692:699 the fragmentation behavior 674:699 the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions 674:803 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 5 54 with ions 1067:1070 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 5 55 theme acid 896:899 arg1 sequence 901:908 their amino acid sequence 884:908 their amino acid sequence 884:908 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 9 56 theme ions 1526:1529 arg1 appearance 1501:1510 The appearance 1497:1510 The appearance of type (iii) ions 1497:1529 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 10 57 with residue 1719:1725 arg1 moiety 1746:1751 intact glycan moiety 1732:1751 intact glycan moiety 1732:1751 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 58 theme intact 1732:1737 arg1 moiety 1746:1751 intact glycan moiety 1732:1751 intact glycan moiety 1732:1751 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 59 gly glycosylated 1634:1645 arg1 located 1654:1660 located 1654:1660 located 1654:1660 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 59 gly glycosylated 1634:1645 arg1 Asn 1647:1649 a glycosylated Asn 1632:1649 a glycosylated Asn 1632:1649 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 11 60 theme Observed 1799:1806 arg1 fragments 1808:1816 Observed fragments 1799:1816 Observed fragments 1799:1816 Observed fragments are reasonably explained by a combination of existing fragmentation rules suggested for N-glycans and peptides. 24664808 5 61 theme fragment 963:970 arg1 ions 972:975 (i) glycan fragment ions 952:975 (i) glycan fragment ions 952:975 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 6 62 dep fragments 1198:1206 arg1 ions 1244:1247 E ions 1242:1247 E ions 1242:1247 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 62 dep fragments 1198:1206 arg1 D 1235:1235 D 1235:1235 D 1235:1235 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 62 dep fragments 1198:1206 arg1 AR-1 1229:1232 AR-1 1229:1232 AR-1 1229:1232 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 62 dep fragments 1198:1206 arg1 AR 1220:1221 (2,4)AR 1215:1221 (2,4)AR 1215:1221 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 63 theme E 1242:1242 arg1 ions 1244:1247 E ions 1242:1247 E ions 1242:1247 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 6 63 theme E 1242:1242 arg1 AR 1220:1221 (2,4)AR 1215:1221 (2,4)AR 1215:1221 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 4 64 attach derived 596:602 arg2 glycopeptides 582:594 glycopeptides 582:594 glycopeptides derived from well-characterized glycoproteins 582:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 64 attach derived 596:602 arg1 glycoproteins 628:640 well-characterized glycoproteins 609:640 well-characterized glycoproteins 609:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 65 gly glycopeptides 582:594 arg2 glycopeptides 582:594 glycopeptides 582:594 glycopeptides derived from well-characterized glycoproteins 582:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 8 66 theme 0,2 1459:1461 arg1 X0-H 1463:1466 (0,2)X0-H 1458:1466 (0,2)X0-H 1458:1466 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 11 67 theme rules 1886:1890 arg1 combination 1848:1858 a combination 1846:1858 a combination of existing fragmentation rules suggested for N-glycans and peptides 1846:1927 Observed fragments are reasonably explained by a combination of existing fragmentation rules suggested for N-glycans and peptides. 24664808 4 68 theme low-energy 746:755 arg1 dissociation 775:786 low-energy collision-induced dissociation 746:786 low-energy collision-induced dissociation (CID) conditions 746:803 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 68 theme low-energy 746:755 arg1 CID 789:791 CID 789:791 CID 789:791 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 11 69 theme existing 1863:1870 arg1 rules 1886:1890 existing fragmentation rules 1863:1890 existing fragmentation rules suggested for N-glycans and peptides 1863:1927 Observed fragments are reasonably explained by a combination of existing fragmentation rules suggested for N-glycans and peptides. 24664808 4 70 theme dissociation 775:786 arg1 conditions 794:803 low-energy collision-induced dissociation (CID) conditions 746:803 low-energy collision-induced dissociation (CID) conditions 746:803 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 9 71 theme Glu 1622:1624 arg1 presence 1596:1603 the presence 1592:1603 the presence of Asp, Asn, and Glu 1592:1624 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 1 72 theme structural 134:143 arg1 analysis 145:152 Glycopeptide structural analysis 121:152 Glycopeptide structural analysis using tandem mass spectrometry 121:183 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 5 73 theme ion 819:821 arg1 species 823:829 The fragment ion species 806:829 The fragment ion species 806:829 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 6 74 theme CID 1103:1105 arg1 spectra 1107:1113 The CID spectra 1099:1113 The CID spectra of glycopeptides having a short peptide sequence 1099:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 3 75 theme protonated 348:357 arg1 glycopeptides 359:371 protonated glycopeptides 348:371 protonated glycopeptides 348:371 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 0 76 theme amino 84:88 arg1 composition 95:105 amino acid composition 84:105 amino acid composition 84:105 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 1 77 theme mass 167:170 arg1 spectrometry 172:183 tandem mass spectrometry 160:183 tandem mass spectrometry 160:183 Glycopeptide structural analysis using tandem mass spectrometry is becoming a common approach for elucidating site-specific N-glycosylation. 24664808 9 78 theme Asn 1613:1615 arg1 presence 1596:1603 the presence 1592:1603 the presence of Asp, Asn, and Glu 1592:1624 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 0 79 theme composition 95:105 arg1 influences 70:79 influences 70:79 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 0:119 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 4 80 gly glycoproteins 628:640 arg1 glycoproteins 628:640 well-characterized glycoproteins 609:640 well-characterized glycoproteins 609:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 5 81 theme glycan-lost 983:993 arg1 ions 1004:1007 glycan-lost fragment ions 983:1007 glycan-lost fragment ions 983:1007 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 4 82 theme glycopeptides 582:594 arg1 sets 574:577 large sets 568:577 large sets of glycopeptides derived from well-characterized glycoproteins 568:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 4 82 theme glycopeptides 582:594 arg1 glycopeptides 582:594 glycopeptides 582:594 glycopeptides derived from well-characterized glycoproteins 582:640 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 0 83 theme sequence 111:118 arg1 influences 70:79 influences 70:79 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 0:119 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 9 84 theme type 1515:1518 arg1 ions 1526:1529 type (iii) ions 1515:1529 type (iii) ions 1515:1529 The appearance of type (iii) ions strongly depended on the peptide sequence, and especially on the presence of Asp, Asn, and Glu. 24664808 8 85 with glycopeptides 1347:1359 arg1 sequences 1389:1397 long peptide sequences 1376:1397 long peptide sequences 1376:1397 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 8 85 with glycopeptides 1347:1359 arg1 medium 1366:1371 medium 1366:1371 medium 1366:1371 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 7 86 theme moiety 1317:1322 arg1 features 1294:1301 detailed structural features 1274:1301 detailed structural features of the glycan moiety such as branching 1274:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 7 86 theme moiety 1317:1322 arg1 branching 1332:1340 branching 1332:1340 branching 1332:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 3 87 theme mode 513:516 arg1 analysis 518:525 negative-ion mode analysis 500:525 negative-ion mode analysis 500:525 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 5 88 theme cleavage 1029:1036 arg1 products 1038:1045 their secondary cleavage products 1013:1045 their secondary cleavage products 1013:1045 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 6 89 theme short 1141:1145 arg1 sequence 1155:1162 a short peptide sequence 1139:1162 a short peptide sequence 1139:1162 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 3 90 theme deprotonated 446:457 arg1 glycopeptides 459:471 deprotonated glycopeptides 446:471 deprotonated glycopeptides 446:471 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 0 91 theme deprotonated 33:44 arg1 glycopeptides 55:67 deprotonated N-linked glycopeptides 33:67 deprotonated N-linked glycopeptides 33:67 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 0 92 dep characteristics 14:28 arg1 influences 70:79 influences 70:79 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 0:119 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 3 93 theme glycopeptide 540:551 arg1 signals 553:559 glycopeptide signals 540:559 glycopeptide signals 540:559 Therefore, fragmentation of protonated glycopeptides has been extensively investigated; however, few studies are available on deprotonated glycopeptides, despite the usefulness of negative-ion mode analysis in detecting glycopeptide signals. 24664808 0 94 theme glycopeptides 55:67 arg1 characteristics 14:28 Fragmentation characteristics 0:28 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 0:119 Fragmentation characteristics of deprotonated N-linked glycopeptides: influences of amino acid composition and sequence. 24664808 5 95 dep ions 1067:1070 arg1 iii 1053:1055 iii 1053:1055 iii 1053:1055 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 7 96 theme detailed 1274:1281 arg1 features 1294:1301 detailed structural features 1274:1301 detailed structural features of the glycan moiety such as branching 1274:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 7 96 theme detailed 1274:1281 arg1 branching 1332:1340 branching 1332:1340 branching 1332:1340 These fragments define detailed structural features of the glycan moiety such as branching. 24664808 10 97 theme interesting 1684:1694 arg1 fragment 1696:1703 an interesting fragment 1681:1703 an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), 1681:1774 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 8 98 theme long 1376:1379 arg1 sequences 1389:1397 long peptide sequences 1376:1397 long peptide sequences 1376:1397 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 6 99 theme glycan 1191:1196 arg1 fragments 1198:1206 type (i) glycan fragments 1182:1206 type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions) 1182:1248 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 5 100 dep ions 972:975 arg1 i 953:953 i 953:953 i 953:953 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 8 101 dep type 1425:1428 arg1 ii 1431:1432 ii 1431:1432 ii 1431:1432 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 8 101 dep type 1425:1428 arg1 ions 1435:1438 ions 1435:1438 ions (e.g., [peptide + (0,2)X0-H] 1435:1467 For glycopeptides with medium or long peptide sequences, the major fragments were type (ii) ions (e.g., [peptide + (0,2)X0-H](-) and [peptide-NH3-H](-)). 24664808 6 102 theme i 1188:1188 arg1 fragments 1198:1206 type (i) glycan fragments 1182:1206 type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions) 1182:1248 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 4 103 theme N-linked 717:724 arg1 glycopeptides 726:738 deprotonated N-linked glycopeptides 704:738 deprotonated N-linked glycopeptides 704:738 Here, large sets of glycopeptides derived from well-characterized glycoproteins were investigated to understand the fragmentation behavior of deprotonated N-linked glycopeptides under low-energy collision-induced dissociation (CID) conditions. 24664808 5 104 with ions 972:975 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 10 105 theme Asn 1715:1717 arg1 Asn-36 1764:1769 Asn-36 1764:1769 Asn-36 1764:1769 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 105 theme Asn 1715:1717 arg1 residue 1719:1725 an Asn residue 1712:1725 an Asn residue with intact glycan moiety 1712:1751 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 105 theme Asn 1715:1717 arg1 [glycan 1754:1760 [glycan 1754:1760 [glycan 1754:1760 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 5 106 theme intact 1077:1082 arg1 moiety 1091:1096 intact glycan moiety 1077:1096 intact glycan moiety 1077:1096 The fragment ion species were found to be significantly variable depending on their amino acid sequence and could be classified into three types: (i) glycan fragment ions, (ii) glycan-lost fragment ions and their secondary cleavage products, and (iii) fragment ions with intact glycan moiety. 24664808 6 107 theme type 1182:1185 arg1 fragments 1198:1206 type (i) glycan fragments 1182:1206 type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions) 1182:1248 The CID spectra of glycopeptides having a short peptide sequence were dominated by type (i) glycan fragments (e.g., (2,4)AR, (2,4)AR-1, D, and E ions). 24664808 10 108 contain having 1705:1710 arg1 fragment 1696:1703 an interesting fragment 1681:1703 an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), 1681:1774 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 108 contain having 1705:1710 arg2 residue 1719:1725 an Asn residue 1712:1725 an Asn residue with intact glycan moiety 1712:1751 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 108 contain having 1705:1710 arg2 Asn-36 1764:1769 Asn-36 1764:1769 Asn-36 1764:1769 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 24664808 10 108 contain having 1705:1710 arg2 [glycan 1754:1760 [glycan 1754:1760 [glycan 1754:1760 When a glycosylated Asn is located on the C-terminus, an interesting fragment having an Asn residue with intact glycan moiety, [glycan + Asn-36](-), was abundantly formed. 28381551 6 0 from site 1177:1180 arg1 introduction 1127:1138 the introduction 1123:1138 the introduction of small hydrophobic residues at the site 1123:1180 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 1 1 theme N-glycan 199:206 arg1 macroheterogeneity 226:243 macroheterogeneity 226:243 macroheterogeneity 226:243 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 1 1 theme N-glycan 199:206 arg1 occupancy 215:223 N-glycan sequon occupancy 199:223 N-glycan sequon occupancy (macroheterogeneity) 199:244 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 0 2 theme N-glycosyltransferase 85:105 arg1 mutant 107:112 an engineered N-glycosyltransferase mutant 71:112 an engineered N-glycosyltransferase mutant 71:112 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 1 3 theme sequon 208:213 arg1 macroheterogeneity 226:243 macroheterogeneity 226:243 macroheterogeneity 226:243 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 1 3 theme sequon 208:213 arg1 occupancy 215:223 N-glycan sequon occupancy 199:223 N-glycan sequon occupancy (macroheterogeneity) 199:244 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 7 4 theme macroheterogeneity 1604:1621 arg1 issue 1595:1599 the issue 1591:1599 the issue of macroheterogeneity in glycoprotein preparation 1591:1649 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 5 from issue 1595:1599 arg1 preparation 1639:1649 glycoprotein preparation 1626:1649 glycoprotein preparation 1626:1649 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 3 6 from glycan 564:569 arg1 chain 586:590 the protein chain 574:590 the protein chain 574:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 5 7 theme residues 1022:1029 arg1 access 1006:1011 the access 1002:1011 the access of bulky residues surrounding the occupied Asn into the active site 1002:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 5 8 theme bulky 1016:1020 arg1 residues 1022:1029 bulky residues 1016:1029 bulky residues surrounding the occupied Asn into the active site 1016:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 7 9 contain carrying 1463:1470 arg2 N-glycans 1492:1500 multiple (up to 10) N-glycans 1472:1500 multiple (up to 10) N-glycans 1472:1500 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 9 contain carrying 1463:1470 arg1 glycoprotein 1450:1461 a homogeneous glycoprotein 1436:1461 a homogeneous glycoprotein carrying multiple (up to 10) N-glycans 1436:1500 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 3 10 theme first 558:562 arg1 glycan 564:569 the first glycan 554:569 the first glycan on the protein chain 554:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 2 11 gly glycoproteins 321:333 arg1 glycoproteins 321:333 well-defined glycoproteins 308:333 well-defined glycoproteins 308:333 However, access to well-defined glycoproteins is always important for both basic research and therapeutic purposes. 28381551 7 12 dep 10 1488:1489 arg1 to 1485:1486 to 1485:1486 to 1485:1486 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 13 gly glycoprotein 1626:1637 arg1 glycoprotein 1626:1637 glycoprotein preparation 1626:1649 glycoprotein preparation 1626:1649 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 3 14 theme protein 578:584 arg1 chain 586:590 the protein chain 574:590 the protein chain 574:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 4 15 theme strict 723:728 arg1 selectivity 730:740 strict selectivity 723:740 strict selectivity toward the residues around the Asn of N-glycosylation sequon 723:801 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 6 16 theme ApNGT 1219:1223 arg1 stringency 1205:1214 the stringency 1201:1214 the stringency of ApNGT 1201:1223 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 4 17 theme synthetic 835:843 arg1 peptides 845:852 synthetic peptides 835:852 synthetic peptides 835:852 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 3 18 theme catalytic 485:493 arg1 properties 495:504 the catalytic properties 481:504 the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain 481:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 7 19 theme glycoprotein 1626:1637 arg1 preparation 1639:1649 glycoprotein preparation 1626:1649 glycoprotein preparation 1626:1649 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 6 20 theme small 1143:1147 arg1 residues 1161:1168 small hydrophobic residues 1143:1168 small hydrophobic residues 1143:1168 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 2 21 theme basic 364:368 arg1 research 370:377 basic research 364:377 basic research 364:377 However, access to well-defined glycoproteins is always important for both basic research and therapeutic purposes. 28381551 0 22 theme glycoprotein 26:37 arg1 Production 0:9 Production 0:9 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 0:113 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 6 23 theme hydrophobic 1149:1159 arg1 residues 1161:1168 small hydrophobic residues 1143:1168 small hydrophobic residues 1143:1168 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 7 24 theme homogeneous 1438:1448 arg1 glycoprotein 1450:1461 a homogeneous glycoprotein 1436:1461 a homogeneous glycoprotein carrying multiple (up to 10) N-glycans 1436:1500 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 1 25 theme occurring 125:133 arg1 N-glycoproteins 135:149 Naturally occurring N-glycoproteins 115:149 Naturally occurring N-glycoproteins 115:149 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 4 26 from pleuropneumoniae 701:716 arg1 ApNGT 621:625 ApNGT 621:625 ApNGT 621:625 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 26 from pleuropneumoniae 701:716 arg1 N-glycosyltransferase 659:679 a newly discovered cytoplasmic N-glycosyltransferase 628:679 a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae 628:716 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 3 27 theme N-glycosyltransferases 509:530 arg1 properties 495:504 the catalytic properties 481:504 the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain 481:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 0 28 theme homogeneous 14:24 arg1 glycoprotein 26:37 homogeneous glycoprotein 14:37 homogeneous glycoprotein with multisite modifications 14:66 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 4 29 theme peptides 845:852 arg1 library 824:830 a small library 816:830 a small library of synthetic peptides 816:852 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 6 30 theme enormous 1252:1259 arg1 improvement 1261:1271 enormous improvement 1252:1271 enormous improvement of glycosylation efficiency against both short peptides and proteins 1252:1340 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 5 31 theme ApNGT 971:975 arg1 Gln-469 959:965 Gln-469 959:965 Gln-469 959:965 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 5 31 theme ApNGT 971:975 arg1 ApNGT 971:975 ApNGT 971:975 ApNGT 971:975 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 5 31 theme ApNGT 971:975 arg1 residue 950:956 a critical residue 939:956 a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site 939:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 1 32 theme glycan 250:255 arg1 microheterogeneity 268:285 microheterogeneity 268:285 microheterogeneity 268:285 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 1 32 theme glycan 250:255 arg1 structure 257:265 glycan structure 250:265 glycan structure (microheterogeneity) 250:286 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 0 33 theme multisite 44:52 arg1 modifications 54:66 multisite modifications 44:66 multisite modifications 44:66 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 4 34 gly N-glycosylation 780:794 arg2 sequon 796:801 N-glycosylation sequon 780:801 N-glycosylation sequon 780:801 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 7 35 theme multiple 1472:1479 arg1 N-glycans 1492:1500 multiple (up to 10) N-glycans 1472:1500 multiple (up to 10) N-glycans 1472:1500 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 36 theme efficient 1369:1377 arg1 Q469A 1387:1391 Q469A 1387:1391 Q469A 1387:1391 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 36 theme efficient 1369:1377 arg1 mutant 1379:1384 the most efficient mutant 1360:1384 the most efficient mutant (Q469A) other than the wild-type ApNGT 1360:1423 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 6 37 theme residues 1161:1168 arg1 introduction 1127:1138 the introduction 1123:1138 the introduction of small hydrophobic residues at the site 1123:1180 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 7 38 gly glycoprotein 1450:1461 arg1 glycoprotein 1450:1461 a homogeneous glycoprotein 1436:1461 a homogeneous glycoprotein carrying multiple (up to 10) N-glycans 1436:1500 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 6 39 theme short 1314:1318 arg1 peptides 1320:1327 both short peptides 1309:1327 both short peptides 1309:1327 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 0 40 with glycoprotein 26:37 arg1 modifications 54:66 multisite modifications 44:66 multisite modifications 44:66 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 3 41 theme substantial 435:445 arg1 effort 447:452 a substantial effort 433:452 a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain 433:590 As a result, there has been a substantial effort to identify and understand the catalytic properties of N-glycosyltransferases, enzymes that install the first glycan on the protein chain. 28381551 1 42 gly N-glycoproteins 135:149 arg1 N-glycoproteins 135:149 Naturally occurring N-glycoproteins 115:149 Naturally occurring N-glycoproteins 115:149 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 0 43 gly glycoprotein 26:37 arg1 glycoprotein 26:37 homogeneous glycoprotein 14:37 homogeneous glycoprotein with multisite modifications 14:66 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 1 44 theme glycoform 159:167 arg1 heterogeneity 169:181 glycoform heterogeneity 159:181 glycoform heterogeneity 159:181 Naturally occurring N-glycoproteins exhibit glycoform heterogeneity with respect to N-glycan sequon occupancy (macroheterogeneity) and glycan structure (microheterogeneity). 28381551 6 45 theme efficiency 1290:1299 arg1 improvement 1261:1271 enormous improvement 1252:1271 enormous improvement of glycosylation efficiency against both short peptides and proteins 1252:1340 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 2 46 theme well-defined 308:319 arg1 glycoproteins 321:333 well-defined glycoproteins 308:333 well-defined glycoproteins 308:333 However, access to well-defined glycoproteins is always important for both basic research and therapeutic purposes. 28381551 6 47 theme Site-saturated 1082:1095 arg1 mutagenesis 1097:1107 Site-saturated mutagenesis 1082:1107 Site-saturated mutagenesis 1082:1107 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 2 48 theme therapeutic 383:393 arg1 purposes 395:402 therapeutic purposes 383:402 therapeutic purposes 383:402 However, access to well-defined glycoproteins is always important for both basic research and therapeutic purposes. 28381551 4 49 theme small 818:822 arg1 library 824:830 a small library 816:830 a small library of synthetic peptides 816:852 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 50 theme discovered 636:645 arg1 ApNGT 621:625 ApNGT 621:625 ApNGT 621:625 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 50 theme discovered 636:645 arg1 N-glycosyltransferase 659:679 a newly discovered cytoplasmic N-glycosyltransferase 628:679 a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae 628:716 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 5 51 theme occupied 1047:1054 arg1 Asn 1056:1058 the occupied Asn 1043:1058 the occupied Asn into the active site 1043:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 4 52 contain has 719:721 arg2 selectivity 730:740 strict selectivity 723:740 strict selectivity toward the residues around the Asn of N-glycosylation sequon 723:801 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 52 contain has 719:721 arg1 ApNGT 621:625 ApNGT 621:625 ApNGT 621:625 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 52 contain has 719:721 arg1 N-glycosyltransferase 659:679 a newly discovered cytoplasmic N-glycosyltransferase 628:679 a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae 628:716 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 6 53 theme glycosylation 1276:1288 arg1 efficiency 1290:1299 glycosylation efficiency 1276:1299 glycosylation efficiency 1276:1299 Site-saturated mutagenesis revealed that the introduction of small hydrophobic residues at the site cannot only weaken the stringency of ApNGT but can also contribute to enormous improvement of glycosylation efficiency against both short peptides and proteins. 28381551 4 54 theme N-glycosylation 780:794 arg1 sequon 796:801 N-glycosylation sequon 780:801 N-glycosylation sequon 780:801 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 55 theme sequon 796:801 arg1 Asn 773:775 the Asn 769:775 the Asn of N-glycosylation sequon 769:801 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 0 56 theme engineered 74:83 arg1 mutant 107:112 an engineered N-glycosyltransferase mutant 71:112 an engineered N-glycosyltransferase mutant 71:112 Production of homogeneous glycoprotein with multisite modifications by an engineered N-glycosyltransferase mutant. 28381551 7 57 theme wild-type 1409:1417 arg1 ApNGT 1419:1423 the wild-type ApNGT 1405:1423 the most efficient mutant (Q469A) other than the wild-type ApNGT 1360:1423 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 4 58 theme Actinobacillus 686:699 arg1 pleuropneumoniae 701:716 Actinobacillus pleuropneumoniae 686:716 Actinobacillus pleuropneumoniae 686:716 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 7 59 dep multiple 1472:1479 arg1 10 1488:1489 10 1488:1489 10 1488:1489 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 5 60 theme critical 941:948 arg1 Gln-469 959:965 Gln-469 959:965 Gln-469 959:965 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 5 60 theme critical 941:948 arg1 ApNGT 971:975 ApNGT 971:975 ApNGT 971:975 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 5 60 theme critical 941:948 arg1 residue 950:956 a critical residue 939:956 a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site 939:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 7 61 dep other 1394:1398 arg1 than 1400:1403 than 1400:1403 than 1400:1403 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 5 62 theme inherent 859:866 arg1 stringency 868:877 The inherent stringency 855:877 The inherent stringency 855:877 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 7 63 theme promising 1542:1550 arg1 construct 1527:1535 this construct 1522:1535 this construct 1522:1535 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 7 63 theme promising 1542:1550 arg1 biocatalyst 1552:1562 a promising biocatalyst 1540:1562 a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation 1540:1649 We then employed the most efficient mutant (Q469A) other than the wild-type ApNGT to produce a homogeneous glycoprotein carrying multiple (up to 10) N-glycans, demonstrating that this construct is a promising biocatalyst for potentially addressing the issue of macroheterogeneity in glycoprotein preparation. 28381551 5 64 theme active 1069:1074 arg1 site 1076:1079 the active site 1065:1079 the active site 1065:1079 The inherent stringency was subsequently demonstrated to be closely associated with a critical residue (Gln-469) of ApNGT which we propose hinders the access of bulky residues surrounding the occupied Asn into the active site. 28381551 4 65 theme cytoplasmic 647:657 arg1 ApNGT 621:625 ApNGT 621:625 ApNGT 621:625 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 28381551 4 65 theme cytoplasmic 647:657 arg1 N-glycosyltransferase 659:679 a newly discovered cytoplasmic N-glycosyltransferase 628:679 a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae 628:716 In this study we found that ApNGT, a newly discovered cytoplasmic N-glycosyltransferase from Actinobacillus pleuropneumoniae, has strict selectivity toward the residues around the Asn of N-glycosylation sequon by screening a small library of synthetic peptides. 25184632 6 0 theme glycoforms 1238:1247 arg1 distribution 1213:1224 Differential tissue distribution 1193:1224 Differential tissue distribution of N-linked glycoforms 1193:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 8 1 theme changes 1591:1597 arg1 analysis 1568:1575 higher throughput analysis 1550:1575 higher throughput analysis of the global changes in N-glycosylation associated with cancers 1550:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 4 2 theme permethylation 976:989 arg1 analysis 991:998 off-tissue permethylation analysis 965:998 off-tissue permethylation analysis combined with multiple database comparisons 965:1042 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 5 3 dep cortex 1173:1178 arg1 the 1169:1171 the 1169:1171 the 1169:1171 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 1 4 from tissues 345:351 arg1 distribution 286:297 distribution 286:297 distribution 286:297 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 4 from tissues 345:351 arg1 profile 261:267 profile the location and distribution 261:297 profile 261:267 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 5 theme multiple 302:309 arg1 species 327:333 multiple N-linked glycan species 302:333 multiple N-linked glycan species in frozen tissues 302:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 4 6 theme structural 924:933 arg1 assignments 935:945 Other structural assignments 918:945 Other structural assignments 918:945 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 7 7 theme MALDI-IMS 1316:1324 arg1 profiling 1333:1341 MALDI-IMS glycan profiling 1316:1341 MALDI-IMS glycan profiling 1316:1341 The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. 25184632 5 8 from distributions 1114:1126 arg1 medulla 1184:1190 medulla 1184:1190 medulla 1184:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 5 8 from distributions 1114:1126 arg1 cortex 1173:1178 cortex 1173:1178 cortex 1173:1178 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 1 9 theme mass 211:214 arg1 spectrometry 216:227 matrix-assisted laser desorption/ionization imaging mass spectrometry 159:227 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 9 theme mass 211:214 arg1 MALDI-IMS 230:238 MALDI-IMS 230:238 MALDI-IMS 230:238 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 10 theme hepatocellular 590:603 arg1 carcinoma 605:613 a human hepatocellular carcinoma 582:613 a human hepatocellular carcinoma tissue microarray 582:631 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 3 11 theme structural 799:808 arg1 composition 810:820 the structural composition 795:820 the structural composition of a subset of glycans 795:843 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 1 12 theme glycans 411:417 arg1 analysis 399:406 the direct analysis 388:406 the direct analysis of glycans 388:417 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 13 with digestion 691:699 arg1 F 728:728 peptide N-glycosidase F 706:728 peptide N-glycosidase F 706:728 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 8 14 contain has 1462:1464 arg2 potential 1470:1478 the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers 1466:1640 the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers 1466:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 8 14 contain has 1462:1464 arg1 workflow 1453:1460 This MALDI-IMS workflow 1438:1460 This MALDI-IMS workflow 1438:1460 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 2 15 from kidney 531:536 arg1 microarray 622:631 a human hepatocellular carcinoma tissue microarray 582:631 a human hepatocellular carcinoma tissue microarray 582:631 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 2 15 from kidney 531:536 arg1 cancers 569:575 human pancreatic and prostate cancers 539:575 cancers 569:575 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 2 15 from kidney 531:536 arg1 tissues 505:511 Formalin-fixed tissues 490:511 Formalin-fixed tissues from normal mouse kidney 490:536 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 3 16 theme glycans 837:843 arg1 subset 827:832 a subset 825:832 a subset of glycans 825:843 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 1 17 from distribution 286:297 arg1 tissues 345:351 frozen tissues 338:351 frozen tissues 338:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 18 link N-linked 1146:1153 arg1 glycoforms 1155:1164 major cellular N-linked glycoforms 1131:1164 major cellular N-linked glycoforms in the cortex and medulla 1131:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 1 19 dep profile 261:267 arg1 location 273:280 the location 269:280 the location 269:280 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 20 theme tissue 615:620 arg1 microarray 622:631 a human hepatocellular carcinoma tissue microarray 582:631 a human hepatocellular carcinoma tissue microarray 582:631 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 3 21 theme subset 827:832 arg1 composition 810:820 the structural composition 795:820 the structural composition of a subset of glycans 795:843 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 1 22 theme formalin-fixed 441:454 arg1 tissues 481:487 formalin-fixed paraffin-embedded (FFPE) tissues 441:487 clinically derived formalin-fixed paraffin-embedded (FFPE) tissues 422:487 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 4 23 theme multiple 1014:1021 arg1 comparisons 1032:1042 multiple database comparisons 1014:1042 multiple database comparisons 1014:1042 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 1 24 from profile 261:267 arg1 tissues 345:351 frozen tissues 338:351 frozen tissues 338:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 0 25 theme formalin-fixed 58:71 arg1 tissue 100:105 formalin-fixed paraffin embedded clinical tissue 58:105 formalin-fixed paraffin embedded clinical tissue 58:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 26 theme matrix-assisted 159:173 arg1 spectrometry 216:227 matrix-assisted laser desorption/ionization imaging mass spectrometry 159:227 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 26 theme matrix-assisted 159:173 arg1 MALDI-IMS 230:238 MALDI-IMS 230:238 MALDI-IMS 230:238 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 27 link derived 433:439 arg1 tissues 481:487 formalin-fixed paraffin-embedded (FFPE) tissues 441:487 clinically derived formalin-fixed paraffin-embedded (FFPE) tissues 422:487 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 0 28 theme embedded 82:89 arg1 tissue 100:105 formalin-fixed paraffin embedded clinical tissue 58:105 formalin-fixed paraffin embedded clinical tissue 58:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 29 theme desorption/ionization 181:201 arg1 spectrometry 216:227 matrix-assisted laser desorption/ionization imaging mass spectrometry 159:227 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 29 theme desorption/ionization 181:201 arg1 MALDI-IMS 230:238 MALDI-IMS 230:238 MALDI-IMS 230:238 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 30 theme N-glycosidase 714:726 arg1 F 728:728 peptide N-glycosidase F 706:728 peptide N-glycosidase F 706:728 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 3 31 theme on-tissue 875:883 arg1 fragmentation 903:915 on-tissue collision-induced fragmentation 875:915 on-tissue collision-induced fragmentation 875:915 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 2 32 theme prostate 560:567 arg1 cancers 569:575 human pancreatic and prostate cancers 539:575 cancers 569:575 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 8 33 from N-glycosylation 1602:1616 arg1 analysis 1568:1575 higher throughput analysis 1550:1575 higher throughput analysis of the global changes in N-glycosylation associated with cancers 1550:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 5 34 theme tissue 1107:1112 arg1 distributions 1114:1126 specific tissue distributions 1098:1126 specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla 1098:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 5 35 gly glycoforms 1155:1164 arg2 cortex 1173:1178 cortex 1173:1178 cortex 1173:1178 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 5 35 gly glycoforms 1155:1164 arg2 medulla 1184:1190 medulla 1184:1190 medulla 1184:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 7 36 theme non-tumor 1369:1377 arg1 tissues 1379:1385 non-tumor tissues 1369:1385 non-tumor tissues 1369:1385 The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. 25184632 8 37 theme tissue 1506:1511 arg1 block 1513:1517 any FFPE tissue block 1497:1517 any FFPE tissue block 1497:1517 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 0 38 theme MALDI 0:4 arg1 spectrometry 19:30 MALDI imaging mass spectrometry profiling 0:40 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue 0:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 39 theme N-linked 311:318 arg1 species 327:333 multiple N-linked glycan species 302:333 multiple N-linked glycan species in frozen tissues 302:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 40 theme mouse 1056:1060 arg1 sections 1076:1083 mouse kidney tissue sections 1056:1083 mouse kidney tissue sections 1056:1083 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 6 41 theme Differential 1193:1204 arg1 distribution 1213:1224 Differential tissue distribution 1193:1224 Differential tissue distribution of N-linked glycoforms 1193:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 0 42 theme mass 14:17 arg1 spectrometry 19:30 MALDI imaging mass spectrometry profiling 0:40 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue 0:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 43 theme species 327:333 arg1 distribution 286:297 distribution 286:297 distribution 286:297 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 43 theme species 327:333 arg1 profile 261:267 profile the location and distribution 261:297 profile 261:267 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 44 theme tissue 1069:1074 arg1 sections 1076:1083 mouse kidney tissue sections 1056:1083 mouse kidney tissue sections 1056:1083 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 2 45 theme mouse 525:529 arg1 kidney 531:536 normal mouse kidney 518:536 normal mouse kidney 518:536 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 1 46 theme imaging 203:209 arg1 spectrometry 216:227 matrix-assisted laser desorption/ionization imaging mass spectrometry 159:227 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 46 theme imaging 203:209 arg1 MALDI-IMS 230:238 MALDI-IMS 230:238 MALDI-IMS 230:238 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 47 theme frozen 338:343 arg1 tissues 345:351 frozen tissues 338:351 frozen tissues 338:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 0 48 theme profiling 32:40 arg1 spectrometry 19:30 MALDI imaging mass spectrometry profiling 0:40 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue 0:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 5 49 theme glycoforms 1155:1164 arg1 distributions 1114:1126 specific tissue distributions 1098:1126 specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla 1098:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 8 50 from changes 1591:1597 arg1 N-glycosylation 1602:1616 N-glycosylation 1602:1616 N-glycosylation associated with cancers 1602:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 0 51 theme N-glycans 45:53 arg1 spectrometry 19:30 MALDI imaging mass spectrometry profiling 0:40 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue 0:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 4 52 theme Other 918:922 arg1 assignments 935:945 Other structural assignments 918:945 Other structural assignments 918:945 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 6 53 link N-linked 1229:1236 arg1 glycoforms 1238:1247 N-linked glycoforms 1229:1247 N-linked glycoforms 1229:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 1 54 dep method 241:246 arg1 spatially 251:259 spatially 251:259 to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 248:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 55 theme cellular 1137:1144 arg1 glycoforms 1155:1164 major cellular N-linked glycoforms 1131:1164 major cellular N-linked glycoforms in the cortex and medulla 1131:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 7 56 theme microarray 1398:1407 arg1 format 1409:1414 a tumor microarray format 1390:1414 a tumor microarray format 1390:1414 The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. 25184632 3 57 theme MALDI-IMS 771:779 arg1 analysis 781:788 MALDI-IMS analysis 771:788 MALDI-IMS analysis 771:788 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 2 58 theme antigen 651:657 arg1 retrieval 659:667 antigen retrieval 651:667 antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F 651:728 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 6 59 theme N-linked 1229:1236 arg1 glycoforms 1238:1247 N-linked glycoforms 1229:1247 N-linked glycoforms 1229:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 4 60 theme off-tissue 965:974 arg1 analysis 991:998 off-tissue permethylation analysis 965:998 off-tissue permethylation analysis combined with multiple database comparisons 965:1042 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 1 61 dep paraffin-embedded 456:472 arg1 FFPE 475:478 FFPE 475:478 FFPE 475:478 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 62 from species 327:333 arg1 tissues 345:351 frozen tissues 338:351 frozen tissues 338:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 63 from cortex 1173:1178 arg1 distributions 1114:1126 specific tissue distributions 1098:1126 specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla 1098:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 2 64 theme on-tissue 681:689 arg1 digestion 691:699 on-tissue digestion 681:699 on-tissue digestion with peptide N-glycosidase F 681:728 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 1 65 theme spectrometry 216:227 arg1 method 241:246 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method 138:246 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 8 66 from analysis 1568:1575 arg1 N-glycosylation 1602:1616 N-glycosylation 1602:1616 N-glycosylation associated with cancers 1602:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 4 67 theme database 1023:1030 arg1 comparisons 1032:1042 multiple database comparisons 1014:1042 multiple database comparisons 1014:1042 Other structural assignments were confirmed by off-tissue permethylation analysis combined with multiple database comparisons. 25184632 8 68 theme global 1584:1589 arg1 changes 1591:1597 the global changes 1580:1597 the global changes in N-glycosylation associated with cancers 1580:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 6 69 theme tissue 1280:1285 arg1 types 1287:1291 the other tissue types 1270:1291 the other tissue types 1270:1291 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 5 70 from glycoforms 1155:1164 arg1 medulla 1184:1190 medulla 1184:1190 medulla 1184:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 5 70 from glycoforms 1155:1164 arg1 cortex 1173:1178 cortex 1173:1178 cortex 1173:1178 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 1 71 theme derived 433:439 arg1 tissues 481:487 formalin-fixed paraffin-embedded (FFPE) tissues 441:487 clinically derived formalin-fixed paraffin-embedded (FFPE) tissues 422:487 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 72 theme paraffin-embedded 456:472 arg1 tissues 481:487 formalin-fixed paraffin-embedded (FFPE) tissues 441:487 clinically derived formalin-fixed paraffin-embedded (FFPE) tissues 422:487 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 73 theme carcinoma 605:613 arg1 microarray 622:631 a human hepatocellular carcinoma tissue microarray 582:631 a human hepatocellular carcinoma tissue microarray 582:631 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 6 74 located observed 1258:1265 arg2 distribution 1213:1224 Differential tissue distribution 1193:1224 Differential tissue distribution of N-linked glycoforms 1193:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 6 74 located observed 1258:1265 arg1 types 1287:1291 the other tissue types 1270:1291 the other tissue types 1270:1291 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 8 75 theme MALDI-IMS 1443:1451 arg1 workflow 1453:1460 This MALDI-IMS workflow 1438:1460 This MALDI-IMS workflow 1438:1460 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 5 76 from medulla 1184:1190 arg1 distributions 1114:1126 specific tissue distributions 1098:1126 specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla 1098:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 1 77 theme developed 149:157 arg1 method 241:246 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method 138:246 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 0 78 from spectrometry 19:30 arg1 tissue 100:105 formalin-fixed paraffin embedded clinical tissue 58:105 formalin-fixed paraffin embedded clinical tissue 58:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 0 79 theme clinical 91:98 arg1 tissue 100:105 formalin-fixed paraffin embedded clinical tissue 58:105 formalin-fixed paraffin embedded clinical tissue 58:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 80 theme laser 175:179 arg1 spectrometry 216:227 matrix-assisted laser desorption/ionization imaging mass spectrometry 159:227 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues 138:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 1 80 theme laser 175:179 arg1 MALDI-IMS 230:238 MALDI-IMS 230:238 MALDI-IMS 230:238 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 81 theme human 584:588 arg1 carcinoma 605:613 a human hepatocellular carcinoma 582:613 a human hepatocellular carcinoma tissue microarray 582:631 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 5 82 theme sections 1076:1083 arg1 Imaging 1045:1051 Imaging 1045:1051 Imaging of mouse kidney tissue sections 1045:1083 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 2 83 theme peptide 706:712 arg1 F 728:728 peptide N-glycosidase F 706:728 peptide N-glycosidase F 706:728 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 3 84 theme collision-induced 885:901 arg1 fragmentation 903:915 on-tissue collision-induced fragmentation 875:915 on-tissue collision-induced fragmentation 875:915 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 0 85 theme tissue 118:123 arg1 microarrays 125:135 and tissue microarrays 114:135 microarrays 125:135 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 1 86 theme direct 392:397 arg1 analysis 399:406 the direct analysis 388:406 the direct analysis of glycans 388:417 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 87 theme specific 1098:1105 arg1 distributions 1114:1126 specific tissue distributions 1098:1126 specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla 1098:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 7 88 theme glycan 1326:1331 arg1 profiling 1333:1341 MALDI-IMS glycan profiling 1316:1341 MALDI-IMS glycan profiling 1316:1341 The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. 25184632 0 89 theme imaging 6:12 arg1 spectrometry 19:30 MALDI imaging mass spectrometry profiling 0:40 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue 0:105 MALDI imaging mass spectrometry profiling of N-glycans in formalin-fixed paraffin embedded clinical tissue blocks and tissue microarrays. 25184632 6 90 theme tissue 1206:1211 arg1 distribution 1213:1224 Differential tissue distribution 1193:1224 Differential tissue distribution of N-linked glycoforms 1193:1247 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 8 91 theme FFPE 1501:1504 arg1 block 1513:1517 any FFPE tissue block 1497:1517 any FFPE tissue block 1497:1517 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 1 92 theme glycan 320:325 arg1 species 327:333 multiple N-linked glycan species 302:333 multiple N-linked glycan species in frozen tissues 302:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 5 93 theme kidney 1062:1067 arg1 sections 1076:1083 mouse kidney tissue sections 1056:1083 mouse kidney tissue sections 1056:1083 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 5 94 theme N-linked 1146:1153 arg1 glycoforms 1155:1164 major cellular N-linked glycoforms 1131:1164 major cellular N-linked glycoforms in the cortex and medulla 1131:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 2 95 theme normal 518:523 arg1 kidney 531:536 normal mouse kidney 518:536 normal mouse kidney 518:536 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 25184632 8 96 theme tissue 1522:1527 arg1 microarray 1529:1538 tissue microarray 1522:1538 tissue microarray 1522:1538 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 6 97 theme other 1274:1278 arg1 types 1287:1291 the other tissue types 1270:1291 the other tissue types 1270:1291 Differential tissue distribution of N-linked glycoforms was also observed in the other tissue types. 25184632 3 98 theme released 735:742 arg1 N-glycans 744:752 The released N-glycans 731:752 The released N-glycans 731:752 The released N-glycans were detected by MALDI-IMS analysis, and the structural composition of a subset of glycans could be verified directly by on-tissue collision-induced fragmentation. 25184632 8 99 theme throughput 1557:1566 arg1 analysis 1568:1575 higher throughput analysis 1550:1575 higher throughput analysis of the global changes in N-glycosylation associated with cancers 1550:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 8 100 theme higher 1550:1555 arg1 analysis 1568:1575 higher throughput analysis 1550:1575 higher throughput analysis of the global changes in N-glycosylation associated with cancers 1550:1640 This MALDI-IMS workflow has the potential to be applied to any FFPE tissue block or tissue microarray to enable higher throughput analysis of the global changes in N-glycosylation associated with cancers. 25184632 5 101 theme major 1131:1135 arg1 glycoforms 1155:1164 major cellular N-linked glycoforms 1131:1164 major cellular N-linked glycoforms in the cortex and medulla 1131:1190 Imaging of mouse kidney tissue sections demonstrates specific tissue distributions of major cellular N-linked glycoforms in the cortex and medulla. 25184632 7 102 theme tumor 1392:1396 arg1 format 1409:1414 a tumor microarray format 1390:1414 a tumor microarray format 1390:1414 The efficacy of using MALDI-IMS glycan profiling to distinguish tumor from non-tumor tissues in a tumor microarray format is also demonstrated. 25184632 1 103 link N-linked 311:318 arg1 species 327:333 multiple N-linked glycan species 302:333 multiple N-linked glycan species in frozen tissues 302:351 A recently developed matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in frozen tissues has been extended and improved for the direct analysis of glycans in clinically derived formalin-fixed paraffin-embedded (FFPE) tissues. 25184632 2 104 theme Formalin-fixed 490:503 arg1 tissues 505:511 Formalin-fixed tissues 490:511 Formalin-fixed tissues from normal mouse kidney 490:536 Formalin-fixed tissues from normal mouse kidney, human pancreatic and prostate cancers, and a human hepatocellular carcinoma tissue microarray were processed by antigen retrieval followed by on-tissue digestion with peptide N-glycosidase F. 26082214 4 0 theme enzymatic 568:576 arg1 treatment 578:586 enzymatic treatment 568:586 enzymatic treatment 568:586 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 3 1 theme homogeneous 405:415 arg1 glycoproteins 417:429 homogeneous glycoproteins 405:429 homogeneous glycoproteins 405:429 Several approaches have been developed to produce homogeneous glycoproteins. 26082214 2 2 theme glycoproteins 328:340 arg1 application 313:323 the application 309:323 the application of glycoproteins in medicine 309:352 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 4 3 link N-linked 487:494 arg1 N-acetylglucosamine 496:514 N-linked N-acetylglucosamine 487:514 N-linked N-acetylglucosamine (GlcNAc) 487:523 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 4 3 link N-linked 487:494 arg1 GlcNAc 517:522 GlcNAc 517:522 GlcNAc 517:522 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 2 4 gly glycoproteins 328:340 arg1 glycoproteins 328:340 glycoproteins 328:340 glycoproteins 328:340 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 5 5 theme N-linked 589:596 arg1 GlcNAc 598:603 N-linked GlcNAc 589:603 N-linked GlcNAc 589:603 N-linked GlcNAc can then be extended by existing methods to produce homogeneous glycoproteins. 26082214 4 6 theme E. 550:551 arg1 cells 558:562 glyco-engineered E. coli cells 533:562 glyco-engineered E. coli cells 533:562 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 0 7 from N-Acetylglucosamine 51:69 arg1 coli 86:89 Escherichia coli 74:89 Escherichia coli 74:89 Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli. 26082214 1 8 gly glycoproteins 148:160 arg1 glycoproteins 148:160 eukaryotic glycoproteins 137:160 eukaryotic glycoproteins 137:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 4 9 contain carrying 478:485 arg2 N-acetylglucosamine 496:514 N-linked N-acetylglucosamine 487:514 N-linked N-acetylglucosamine (GlcNAc) 487:523 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 4 9 contain carrying 478:485 arg2 GlcNAc 517:522 GlcNAc 517:522 GlcNAc 517:522 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 4 9 contain carrying 478:485 arg1 glycoproteins 464:476 glycoproteins 464:476 glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment 464:586 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 4 10 theme N-linked 487:494 arg1 N-acetylglucosamine 496:514 N-linked N-acetylglucosamine 487:514 N-linked N-acetylglucosamine (GlcNAc) 487:523 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 4 10 theme N-linked 487:494 arg1 GlcNAc 517:522 GlcNAc 517:522 GlcNAc 517:522 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 1 11 theme asparagine 110:119 arg1 residues 125:132 asparagine (N) residues 110:132 asparagine (N) residues of eukaryotic glycoproteins 110:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 1 11 theme asparagine 110:119 arg1 glycoproteins 148:160 eukaryotic glycoproteins 137:160 eukaryotic glycoproteins 137:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 5 12 theme homogeneous 657:667 arg1 glycoproteins 669:681 homogeneous glycoproteins 657:681 homogeneous glycoproteins 657:681 N-linked GlcNAc can then be extended by existing methods to produce homogeneous glycoproteins. 26082214 3 13 gly glycoproteins 417:429 arg1 glycoproteins 417:429 homogeneous glycoproteins 405:429 homogeneous glycoproteins 405:429 Several approaches have been developed to produce homogeneous glycoproteins. 26082214 0 14 theme Glycoproteins 14:26 arg1 Production 0:9 Production 0:9 Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli. 0:90 Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli. 26082214 1 15 attach linked 100:105 arg1 residues 125:132 asparagine (N) residues 110:132 asparagine (N) residues of eukaryotic glycoproteins 110:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 1 15 attach linked 100:105 arg1 glycoproteins 148:160 eukaryotic glycoproteins 137:160 eukaryotic glycoproteins 137:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 1 15 attach linked 100:105 arg2 Glycans 92:98 Glycans 92:98 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins 92:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 5 16 link N-linked 589:596 arg1 GlcNAc 598:603 N-linked GlcNAc 589:603 N-linked GlcNAc 589:603 N-linked GlcNAc can then be extended by existing methods to produce homogeneous glycoproteins. 26082214 4 17 dep E. 550:551 arg1 coli 553:556 coli 553:556 coli 553:556 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 2 18 theme particular 268:277 arg1 structures 286:295 particular glycan structures 268:295 particular glycan structures 268:295 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 0 19 theme Asparagine-Linked 33:49 arg1 N-Acetylglucosamine 51:69 Asparagine-Linked N-Acetylglucosamine 33:69 Asparagine-Linked N-Acetylglucosamine in Escherichia coli 33:89 Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli. 26082214 4 20 theme glyco-engineered 533:548 arg1 cells 558:562 glyco-engineered E. coli cells 533:562 glyco-engineered E. coli cells 533:562 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 26082214 3 21 theme Several 355:361 arg1 approaches 363:372 Several approaches 355:372 Several approaches 355:372 Several approaches have been developed to produce homogeneous glycoproteins. 26082214 2 22 theme biological 231:240 arg1 functions 242:250 biological functions 231:250 biological functions associated with particular glycan structures 231:295 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 2 23 theme functions 242:250 arg1 study 222:226 the study 218:226 the study of biological functions associated with particular glycan structures 218:295 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 2 24 from application 313:323 arg1 medicine 345:352 medicine 345:352 medicine 345:352 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 0 25 with Production 0:9 arg1 N-Acetylglucosamine 51:69 Asparagine-Linked N-Acetylglucosamine 33:69 Asparagine-Linked N-Acetylglucosamine in Escherichia coli 33:89 Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli. 26082214 1 26 theme eukaryotic 137:146 arg1 glycoproteins 148:160 eukaryotic glycoproteins 137:160 eukaryotic glycoproteins 137:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 1 27 theme glycoproteins 148:160 arg1 residues 125:132 asparagine (N) residues 110:132 asparagine (N) residues of eukaryotic glycoproteins 110:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 1 27 theme glycoproteins 148:160 arg1 glycoproteins 148:160 eukaryotic glycoproteins 137:160 eukaryotic glycoproteins 137:160 Glycans linked to asparagine (N) residues of eukaryotic glycoproteins are typically heterogeneous. 26082214 5 28 gly glycoproteins 669:681 arg1 glycoproteins 669:681 homogeneous glycoproteins 657:681 homogeneous glycoproteins 657:681 N-linked GlcNAc can then be extended by existing methods to produce homogeneous glycoproteins. 26082214 2 29 theme glycan 279:284 arg1 structures 286:295 particular glycan structures 268:295 particular glycan structures 268:295 This diversity complicates the study of biological functions associated with particular glycan structures and impairs the application of glycoproteins in medicine. 26082214 4 30 gly glycoproteins 464:476 arg1 glycoproteins 464:476 glycoproteins 464:476 glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment 464:586 We describe a method to produce glycoproteins carrying N-linked N-acetylglucosamine (GlcNAc) through glyco-engineered E. coli cells and enzymatic treatment. 27653286 6 0 theme tetramer 1167:1174 arg1 epitopes 1100:1107 The binding epitopes 1088:1107 The binding epitopes 1088:1107 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 0 theme tetramer 1167:1174 arg1 placement 1147:1155 binding site placement 1134:1155 binding site placement 1134:1155 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 0 theme tetramer 1167:1174 arg1 conformation 1116:1127 bound conformation 1110:1127 bound conformation 1110:1127 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 3 1 theme used 449:452 arg1 hosts 464:468 commonly used bacterial hosts 440:468 commonly used bacterial hosts 440:468 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 2 2 theme family 225:230 arg1 interaction 155:165 Its interaction 151:165 Its interaction with heparan sulfate (HS) 151:191 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 2 theme family 225:230 arg1 members 197:203 members 197:203 members of the Slit protein family 197:230 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 8 3 theme site 1602:1605 arg1 examination 1575:1585 examination 1575:1585 examination of the binding site in the context of a previous Robo-Slit structure 1575:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 4 theme glycosylated 754:765 arg1 Robo1-Ig1-2 774:784 Robo1-Ig1-2 774:784 Robo1-Ig1-2 774:784 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 4 theme glycosylated 754:765 arg1 Robo1 767:771 glycosylated Robo1 754:771 glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 754:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 5 theme binding 1134:1140 arg1 placement 1147:1155 binding site placement 1134:1155 binding site placement 1134:1155 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 5 6 from K81 932:934 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 5 6 from K81 932:934 arg1 crosspeak 917:925 the crosspeak 913:925 the crosspeak from K81 on titration with the tetramer 913:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 3 7 theme forms 510:514 arg1 expression 474:483 expression 474:483 expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies 474:580 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 5 8 from crosspeak 917:925 arg1 titration 939:947 titration 939:947 titration with the tetramer 939:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 4 9 theme IdoA-GlcNS6S-IdoA2S-GlcNS6S- 829:856 arg1 tetramer 819:826 a synthetic HS tetramer 804:826 a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 804:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 9 theme IdoA-GlcNS6S-IdoA2S-GlcNS6S- 829:856 arg1 5NH2 862:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 10 theme saturation 1211:1220 arg1 STD 1243:1245 STD 1243:1245 STD 1243:1245 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 10 theme saturation 1211:1220 arg1 difference 1231:1240 saturation transfer difference 1211:1240 saturation transfer difference (STD) 1211:1246 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 11 theme perturbation 1313:1324 arg1 experiments 1326:1336 paramagnetic perturbation experiments 1300:1336 paramagnetic perturbation experiments 1300:1336 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 8 12 theme previous 1627:1634 arg1 structure 1646:1654 a previous Robo-Slit structure 1625:1654 a previous Robo-Slit structure 1625:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 13 gly glycosylated 754:765 arg1 Robo1-Ig1-2 774:784 Robo1-Ig1-2 774:784 Robo1-Ig1-2 774:784 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 13 gly glycosylated 754:765 arg1 Robo1 767:771 glycosylated Robo1 754:771 glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 754:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 8 14 theme structure 1646:1654 arg1 context 1614:1620 the context 1610:1620 the context of a previous Robo-Slit structure 1610:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 1 15 theme cell 91:94 arg1 1 73:73 1 73:73 1 73:73 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 15 theme cell 91:94 arg1 Robo1 79:83 Robo1 79:83 Robo1 79:83 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 15 theme cell 91:94 arg1 molecule 114:121 a cell surface signaling molecule 89:121 a cell surface signaling molecule important in axon guidance 89:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 4 16 theme synthetic 806:814 arg1 tetramer 819:826 a synthetic HS tetramer 804:826 a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 804:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 16 theme synthetic 806:814 arg1 5NH2 862:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 2 17 theme protein 217:223 arg1 family 225:230 the Slit protein family 208:230 the Slit protein family 208:230 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 18 theme Slit 212:215 arg1 family 225:230 the Slit protein family 208:230 the Slit protein family 208:230 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 1 19 theme signaling 104:112 arg1 1 73:73 1 73:73 1 73:73 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 19 theme signaling 104:112 arg1 Robo1 79:83 Robo1 79:83 Robo1 79:83 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 19 theme signaling 104:112 arg1 molecule 114:121 a cell surface signaling molecule 89:121 a cell surface signaling molecule important in axon guidance 89:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 6 20 theme binding 1092:1098 arg1 epitopes 1100:1107 The binding epitopes 1088:1107 The binding epitopes 1088:1107 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 5 21 theme binding 1014:1020 arg1 site 1022:1025 a binding site 1012:1025 a binding site 1012:1025 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 1 22 from guidance 141:148 arg1 important 123:131 important 123:131 important 123:131 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 23 theme important 123:131 arg1 1 73:73 1 73:73 1 73:73 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 23 theme important 123:131 arg1 Robo1 79:83 Robo1 79:83 Robo1 79:83 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 23 theme important 123:131 arg1 molecule 114:121 a cell surface signaling molecule 89:121 a cell surface signaling molecule important in axon guidance 89:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 6 24 theme nuclear 1261:1267 arg1 trNOE 1288:1292 trNOE 1288:1292 trNOE 1288:1292 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 24 theme nuclear 1261:1267 arg1 effect 1280:1285 transferred nuclear Overhauser effect 1249:1285 transferred nuclear Overhauser effect (trNOE) 1249:1293 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 1 25 theme axon 136:139 arg1 guidance 141:148 axon guidance 136:148 axon guidance 136:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 4 26 theme CH2 858:860 arg1 tetramer 819:826 a synthetic HS tetramer 804:826 a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 804:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 26 theme CH2 858:860 arg1 5NH2 862:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 8 27 theme interactions 1705:1716 arg1 modulation 1681:1690 modulation 1681:1690 modulation of Robo-Slit interactions by HS 1681:1722 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 28 theme high 678:681 arg1 content 694:700 high structural content 678:700 high structural content 678:700 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 5 29 theme chemical 881:888 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 8 30 from examination 1575:1585 arg1 context 1614:1620 the context 1610:1620 the context of a previous Robo-Slit structure 1610:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 0 31 theme Structural 0:9 arg1 Aspects 11:17 Structural Aspects 0:17 Structural Aspects of Heparan Sulfate 0:36 Structural Aspects of Heparan Sulfate Binding to Robo1-Ig1-2. 27653286 5 32 theme μM 1060:1061 arg1 constant 1078:1085 a 255 μM disassociation constant 1054:1085 a 255 μM disassociation constant 1054:1085 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 4 33 theme amino 658:662 arg1 type 669:672 a single amino acid type 649:672 a single amino acid type 649:672 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 8 34 from site 1602:1605 arg1 context 1614:1620 the context 1610:1620 the context of a previous Robo-Slit structure 1610:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 6 35 theme paramagnetic 1300:1311 arg1 experiments 1326:1336 paramagnetic perturbation experiments 1300:1336 paramagnetic perturbation experiments 1300:1336 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 5 36 theme constant 1078:1085 arg1 determination 1037:1049 determination 1037:1049 determination of a 255 μM disassociation constant 1037:1085 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 7 37 theme NMR 1424:1426 arg1 experiments 1428:1438 the various NMR experiments 1412:1438 the various NMR experiments 1412:1438 A model of the complex has been generated using constraints derived from the various NMR experiments. 27653286 8 38 theme energetic 1456:1464 arg1 analysis 1466:1473 Postprocessing energetic analysis 1441:1473 Postprocessing energetic analysis of this model 1441:1487 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 3 39 with forms 510:514 arg1 13C 538:540 13C 538:540 13C 538:540 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 3 39 with forms 510:514 arg1 15N 533:535 the uniform 15N 521:535 the uniform 15N 521:535 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 3 39 with forms 510:514 arg1 labeling 550:557 2H labeling 547:557 2H labeling 547:557 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 3 40 gly glycosylated 396:407 arg1 protein 409:415 a glycosylated protein 394:415 a glycosylated protein 394:415 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 3 40 gly glycosylated 396:407 arg1 Robo1 385:389 Robo1 385:389 Robo1 385:389 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 0 41 theme Sulfate 30:36 arg1 Aspects 11:17 Structural Aspects 0:17 Structural Aspects of Heparan Sulfate 0:36 Structural Aspects of Heparan Sulfate Binding to Robo1-Ig1-2. 27653286 2 42 theme interactions 295:306 arg1 characterization 269:284 characterization 269:284 characterization of these interactions by structural methods 269:328 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 8 43 theme binding 1594:1600 arg1 site 1602:1605 the binding site 1590:1605 the binding site in the context of a previous Robo-Slit structure 1590:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 8 44 theme model 1483:1487 arg1 analysis 1466:1473 Postprocessing energetic analysis 1441:1473 Postprocessing energetic analysis of this model 1441:1487 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 3 45 theme bacterial 454:462 arg1 hosts 464:468 commonly used bacterial hosts 440:468 commonly used bacterial hosts 440:468 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 6 46 theme Overhauser 1269:1278 arg1 trNOE 1288:1292 trNOE 1288:1292 trNOE 1288:1292 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 46 theme Overhauser 1269:1278 arg1 effect 1280:1285 transferred nuclear Overhauser effect 1249:1285 transferred nuclear Overhauser effect (trNOE) 1249:1293 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 2 47 theme structural 311:320 arg1 methods 322:328 structural methods 311:328 structural methods 311:328 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 7 48 theme complex 1354:1360 arg1 model 1341:1345 A model 1339:1345 A model of the complex 1339:1360 A model of the complex has been generated using constraints derived from the various NMR experiments. 27653286 4 49 theme Robo1 767:771 arg1 construct 741:749 a two-domain construct 728:749 a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 728:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 50 theme HS 1164:1165 arg1 tetramer 1167:1174 the HS tetramer 1160:1174 the HS tetramer 1160:1174 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 8 51 theme glycan 1528:1533 arg1 residue 1535:1541 each glycan residue 1523:1541 each glycan residue 1523:1541 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 52 theme two-domain 730:739 arg1 construct 741:749 a two-domain construct 728:749 a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 728:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 53 theme site 1142:1145 arg1 placement 1147:1155 binding site placement 1134:1155 binding site placement 1134:1155 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 5 54 from perturbations 896:908 arg1 titration 939:947 titration 939:947 titration with the tetramer 939:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 5 54 from perturbations 896:908 arg1 K81 932:934 K81 932:934 K81 932:934 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 3 55 theme glycosylated 497:508 arg1 forms 510:514 properly glycosylated forms 488:514 properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies 488:580 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 2 56 with members 197:203 arg1 HS 189:190 HS 189:190 HS 189:190 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 56 with members 197:203 arg1 sulfate 180:186 heparan sulfate 172:186 heparan sulfate (HS) 172:191 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 4 57 theme HS 816:817 arg1 tetramer 819:826 a synthetic HS tetramer 804:826 a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2) 804:866 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 57 theme HS 816:817 arg1 5NH2 862:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2 829:865 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 58 theme transfer 1222:1229 arg1 STD 1243:1245 STD 1243:1245 STD 1243:1245 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 58 theme transfer 1222:1229 arg1 difference 1231:1240 saturation transfer difference 1211:1240 saturation transfer difference (STD) 1211:1246 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 5 59 theme initial 975:981 arg1 evidence 983:990 initial evidence 975:990 initial evidence for the location of a binding site 975:1025 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 3 60 theme uniform 525:531 arg1 15N 533:535 the uniform 15N 521:535 the uniform 15N 521:535 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 8 61 dep role 1518:1521 arg1 plays 1543:1547 plays 1543:1547 plays in the binding event 1543:1568 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 6 62 theme bound 1110:1114 arg1 conformation 1116:1127 bound conformation 1110:1127 bound conformation 1110:1127 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 3 63 theme 2H 547:548 arg1 labeling 550:557 2H labeling 547:557 2H labeling 547:557 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 8 64 theme Robo-Slit 1636:1644 arg1 structure 1646:1654 a previous Robo-Slit structure 1625:1654 a previous Robo-Slit structure 1625:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 1 65 theme surface 96:102 arg1 1 73:73 1 73:73 1 73:73 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 65 theme surface 96:102 arg1 Robo1 79:83 Robo1 79:83 Robo1 79:83 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 1 65 theme surface 96:102 arg1 molecule 114:121 a cell surface signaling molecule 89:121 a cell surface signaling molecule important in axon guidance 89:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 3 66 gly glycosylated 497:508 arg1 forms 510:514 properly glycosylated forms 488:514 properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies 488:580 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 5 67 from titration 939:947 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 5 68 theme site 1022:1025 arg1 location 1000:1007 the location 996:1007 the location of a binding site 996:1025 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 4 69 theme alternative 601:611 arg1 methodology 613:623 an alternative methodology 598:623 an alternative methodology 598:623 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 6 70 theme transferred 1249:1259 arg1 trNOE 1288:1292 trNOE 1288:1292 trNOE 1288:1292 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 6 70 theme transferred 1249:1259 arg1 effect 1280:1285 transferred nuclear Overhauser effect 1249:1285 transferred nuclear Overhauser effect (trNOE) 1249:1293 The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. 27653286 8 71 from context 1614:1620 arg1 examination 1575:1585 examination 1575:1585 examination of the binding site in the context of a previous Robo-Slit structure 1575:1654 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 72 dep methodology 613:623 arg1 data 706:709 NMR data 702:709 NMR data 702:709 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 73 theme acid 664:667 arg1 type 669:672 a single amino acid type 649:672 a single amino acid type 649:672 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 1 74 from important 123:131 arg1 guidance 141:148 axon guidance 136:148 axon guidance 136:148 Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. 27653286 5 75 theme Significant 869:879 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 5 76 with titration 939:947 arg1 tetramer 958:965 the tetramer 954:965 the tetramer 954:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 7 77 attach derived 1399:1405 arg2 constraints 1387:1397 constraints 1387:1397 constraints derived from the various NMR experiments 1387:1438 A model of the complex has been generated using constraints derived from the various NMR experiments. 27653286 7 77 attach derived 1399:1405 arg1 experiments 1428:1438 the various NMR experiments 1412:1438 the various NMR experiments 1412:1438 A model of the complex has been generated using constraints derived from the various NMR experiments. 27653286 8 78 theme Postprocessing 1441:1454 arg1 analysis 1466:1473 Postprocessing energetic analysis 1441:1473 Postprocessing energetic analysis of this model 1441:1487 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 0 79 theme Heparan 22:28 arg1 Sulfate 30:36 Heparan Sulfate 22:36 Heparan Sulfate 22:36 Structural Aspects of Heparan Sulfate Binding to Robo1-Ig1-2. 27653286 5 80 theme disassociation 1063:1076 arg1 constant 1078:1085 a 255 μM disassociation constant 1054:1085 a 255 μM disassociation constant 1054:1085 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 8 81 theme Robo-Slit 1695:1703 arg1 interactions 1705:1716 Robo-Slit interactions 1695:1716 Robo-Slit interactions 1695:1716 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 3 82 theme NMR 570:572 arg1 studies 574:580 NMR studies 570:580 NMR studies 570:580 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 4 83 theme single 651:656 arg1 type 669:672 a single amino acid type 649:672 a single amino acid type 649:672 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 3 84 theme glycosylated 396:407 arg1 protein 409:415 a glycosylated protein 394:415 a glycosylated protein 394:415 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 3 84 theme glycosylated 396:407 arg1 Robo1 385:389 Robo1 385:389 Robo1 385:389 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 27653286 5 85 theme crosspeak 917:925 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 7 86 theme various 1416:1422 arg1 experiments 1428:1438 the various NMR experiments 1412:1438 the various NMR experiments 1412:1438 A model of the complex has been generated using constraints derived from the various NMR experiments. 27653286 5 87 theme 255 1056:1058 arg1 constant 1078:1085 a 255 μM disassociation constant 1054:1085 a 255 μM disassociation constant 1054:1085 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 5 88 theme shift 890:894 arg1 perturbations 896:908 Significant chemical shift perturbations 869:908 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer 869:965 Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. 27653286 2 89 with interaction 155:165 arg1 HS 189:190 HS 189:190 HS 189:190 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 89 with interaction 155:165 arg1 sulfate 180:186 heparan sulfate 172:186 heparan sulfate (HS) 172:191 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 90 theme heparan 172:178 arg1 HS 189:190 HS 189:190 HS 189:190 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 2 90 theme heparan 172:178 arg1 sulfate 180:186 heparan sulfate 172:186 heparan sulfate (HS) 172:191 Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. 27653286 4 91 theme structural 683:692 arg1 content 694:700 high structural content 678:700 high structural content 678:700 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 8 92 theme binding 1556:1562 arg1 event 1564:1568 the binding event 1552:1568 the binding event 1552:1568 Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS. 27653286 4 93 with labeling 635:642 arg1 type 669:672 a single amino acid type 649:672 a single amino acid type 649:672 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 4 94 theme NMR 702:704 arg1 data 706:709 NMR data 702:709 NMR data 702:709 Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1-Ig1-2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). 27653286 3 95 theme hosts 464:468 arg1 employment 426:435 employment 426:435 employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies 426:580 However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. 26254006 6 0 theme stress 1083:1088 arg1 resistance 1090:1099 enhanced oxidative stress resistance 1064:1099 enhanced oxidative stress resistance 1064:1099 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 4 1 theme functional 709:718 arg1 Gnt2 720:723 functional Gnt2 709:723 functional Gnt2 709:723 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 2 2 theme fungal 409:414 arg1 pathogens 416:424 several fungal pathogens 401:424 several fungal pathogens including Fusarium oxysporum 401:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 2 2 theme fungal 409:414 arg1 oxysporum 445:453 Fusarium oxysporum 436:453 Fusarium oxysporum 436:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 6 3 theme enhanced 1064:1071 arg1 resistance 1090:1099 enhanced oxidative stress resistance 1064:1099 enhanced oxidative stress resistance 1064:1099 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 3 4 gly hypo-N-glycosylation 594:613 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 0 5 theme Comparative 0:10 arg1 analyses 22:29 Comparative proteomic analyses 0:29 Comparative proteomic analyses 0:29 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 6 6 theme ∆gnt2 1040:1044 arg1 mutant 1046:1051 ∆gnt2 mutant 1040:1051 ∆gnt2 mutant 1040:1051 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 7 7 theme antifungal 1351:1360 arg1 compounds 1362:1370 antifungal compounds 1351:1370 antifungal compounds 1351:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 6 8 from Hypo-N-glycosylation 1016:1035 arg1 mutant 1046:1051 ∆gnt2 mutant 1040:1051 ∆gnt2 mutant 1040:1051 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 7 9 theme trafficking 1241:1251 arg1 vesicles 1253:1260 trafficking vesicles 1241:1260 trafficking vesicles 1241:1260 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 2 10 theme important 348:356 arg1 process 358:364 an important process 345:364 an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum 345:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 2 10 theme important 348:356 arg1 N-glycosylation 306:320 N-glycosylation 306:320 N-glycosylation 306:320 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 5 11 theme wild-type 985:993 arg1 rates 1009:1013 wild-type hyphal fusion rates 985:1013 wild-type hyphal fusion rates 985:1013 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 26254006 6 12 theme proteins 1123:1130 arg1 levels 1113:1118 reduced levels 1105:1118 reduced levels of proteins involved in cell wall organization, biogenesis and remodelling 1105:1193 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 6 12 theme proteins 1123:1130 arg1 resistance 1090:1099 enhanced oxidative stress resistance 1064:1099 enhanced oxidative stress resistance 1064:1099 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 0 13 from glycans 91:97 arg1 oxysporum 131:139 Fusarium oxysporum 122:139 Fusarium oxysporum 122:139 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 4 14 theme wall 775:778 arg1 glycans 780:786 cell wall glycans 770:786 cell wall glycans 770:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 5 15 theme fusion 1002:1007 arg1 rates 1009:1013 wild-type hyphal fusion rates 985:1013 wild-type hyphal fusion rates 985:1013 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 26254006 8 16 theme relative 1658:1665 arg1 abundance 1667:1675 relative abundance 1658:1675 relative abundance 1658:1675 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 8 17 theme Gnt2-mediated 1500:1512 arg1 N-glycosylation 1514:1528 Gnt2-mediated N-glycosylation 1500:1528 Gnt2-mediated N-glycosylation 1500:1528 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 7 18 theme gnt2 1208:1211 arg1 gene 1213:1216 gnt2 gene 1208:1216 gnt2 gene 1208:1216 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 19 theme degrees 855:861 arg1 range 831:835 a broad range 823:835 a broad range of polymerization degrees and differential protein glycosylation patterns 823:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 0 20 theme cell 81:84 arg1 glycans 91:97 cell wall glycans 81:97 cell wall glycans 81:97 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 4 21 theme broad 825:829 arg1 range 831:835 a broad range 823:835 a broad range of polymerization degrees and differential protein glycosylation patterns 823:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 7 22 theme walls 1402:1406 arg1 secretion 1286:1294 secretion 1286:1294 secretion of extracellular proteins related to detoxification of antifungal compounds 1286:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 7 22 theme walls 1402:1406 arg1 degradation 1376:1386 degradation 1376:1386 degradation of plant cell walls 1376:1406 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 1 23 theme post-translational 182:199 arg1 modification 201:212 a ubiquitous post-translational modification 169:212 a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization 169:303 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 1 23 theme post-translational 182:199 arg1 N-glycosylation 150:164 Protein N-glycosylation 142:164 Protein N-glycosylation 142:164 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 2 24 from morphogenesis 370:382 arg1 pathogens 416:424 several fungal pathogens 401:424 several fungal pathogens including Fusarium oxysporum 401:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 2 24 from morphogenesis 370:382 arg1 oxysporum 445:453 Fusarium oxysporum 436:453 Fusarium oxysporum 436:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 7 25 theme cell 1397:1400 arg1 walls 1402:1406 plant cell walls 1391:1406 plant cell walls 1391:1406 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 0 26 theme protein 103:109 arg1 content 111:117 protein content 103:117 protein content in Fusarium oxysporum 103:139 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 6 27 theme wall 1149:1152 arg1 organization 1154:1165 cell wall organization 1144:1165 cell wall organization 1144:1165 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 0 28 theme Gnt2-mediated 43:55 arg1 N-glycosylation 57:71 Gnt2-mediated N-glycosylation 43:71 Gnt2-mediated N-glycosylation 43:71 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 8 29 from abundance 1667:1675 arg1 oxysporum 1683:1691 F. oxysporum 1680:1691 F. oxysporum 1680:1691 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 4 30 theme differential 867:878 arg1 patterns 902:909 differential protein glycosylation patterns 867:909 differential protein glycosylation patterns 867:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 3 31 theme proteome-based 499:512 arg1 analyses 514:521 comparative chemical and proteome-based analyses 474:521 comparative chemical and proteome-based analyses 474:521 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 7 32 theme compounds 1362:1370 arg1 detoxification 1333:1346 detoxification 1333:1346 detoxification of antifungal compounds 1333:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 33 theme glycosylation 888:900 arg1 patterns 902:909 differential protein glycosylation patterns 867:909 differential protein glycosylation patterns 867:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 3 34 theme physiological 548:560 arg1 changes 562:568 the physiological changes 544:568 the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 544:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 3 35 theme chemical 486:493 arg1 analyses 514:521 comparative chemical and proteome-based analyses 474:521 comparative chemical and proteome-based analyses 474:521 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 7 36 from tips 1272:1275 arg1 accumulation 1225:1236 accumulation 1225:1236 accumulation of trafficking vesicles at hyphal tips 1225:1275 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 3 37 theme protein 586:592 arg1 hypo-N-glycosylation 594:613 protein hypo-N-glycosylation 586:613 protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 586:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 5 38 theme normal 945:950 arg1 size 961:964 normal conidium size 945:964 normal conidium size 945:964 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 26254006 8 39 theme protein 1626:1632 arg1 function 1634:1641 protein function 1626:1641 protein function 1626:1641 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 3 40 theme oxysporum 621:629 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 5 41 theme Functional 912:921 arg1 Gnt2 923:926 Functional Gnt2 912:926 Functional Gnt2 912:926 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 26254006 6 42 theme oxidative 1073:1081 arg1 resistance 1090:1099 enhanced oxidative stress resistance 1064:1099 enhanced oxidative stress resistance 1064:1099 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 2 43 from virulence 388:396 arg1 pathogens 416:424 several fungal pathogens 401:424 several fungal pathogens including Fusarium oxysporum 401:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 2 43 from virulence 388:396 arg1 oxysporum 445:453 Fusarium oxysporum 436:453 Fusarium oxysporum 436:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 7 44 theme extracellular 1421:1433 arg1 activity 1453:1460 extracellular polygalacturonase activity 1421:1460 extracellular polygalacturonase activity 1421:1460 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 8 45 theme crucial 1538:1544 arg1 role 1546:1549 a crucial role 1536:1549 a crucial role 1536:1549 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 4 46 theme Gnt2 720:723 arg1 lack 701:704 lack 701:704 lack of functional Gnt2 701:723 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 2 47 theme several 401:407 arg1 pathogens 416:424 several fungal pathogens 401:424 several fungal pathogens including Fusarium oxysporum 401:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 2 47 theme several 401:407 arg1 oxysporum 445:453 Fusarium oxysporum 436:453 Fusarium oxysporum 436:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 7 48 theme hyphal 1265:1270 arg1 tips 1272:1275 hyphal tips 1265:1275 hyphal tips 1265:1275 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 3 49 theme Gnt2-deficient 653:666 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 7 50 theme proteins 1313:1320 arg1 secretion 1286:1294 secretion 1286:1294 secretion of extracellular proteins related to detoxification of antifungal compounds 1286:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 7 50 theme proteins 1313:1320 arg1 degradation 1376:1386 degradation 1376:1386 degradation of plant cell walls 1376:1406 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 51 with polysaccharides 802:816 arg1 range 831:835 a broad range 823:835 a broad range of polymerization degrees and differential protein glycosylation patterns 823:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 7 52 theme vesicles 1253:1260 arg1 accumulation 1225:1236 accumulation 1225:1236 accumulation of trafficking vesicles at hyphal tips 1225:1275 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 53 from glycans 780:786 arg1 size 736:739 the size 732:739 the size of galactofuranose chains in cell wall glycans 732:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 6 54 theme cell 1144:1147 arg1 organization 1154:1165 cell wall organization 1144:1165 cell wall organization 1144:1165 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 3 55 from hypo-N-glycosylation 594:613 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 8 56 from transport 1644:1652 arg1 oxysporum 1683:1691 F. oxysporum 1680:1691 F. oxysporum 1680:1691 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 8 57 from function 1634:1641 arg1 oxysporum 1683:1691 F. oxysporum 1680:1691 F. oxysporum 1680:1691 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 1 58 theme appropriate 234:244 arg1 folding 254:260 appropriate protein folding 234:260 appropriate protein folding 234:260 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 4 59 theme chains 760:765 arg1 size 736:739 the size 732:739 the size of galactofuranose chains in cell wall glycans 732:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 4 60 from chains 760:765 arg1 glycans 780:786 cell wall glycans 770:786 cell wall glycans 770:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 5 61 theme hyphal 995:1000 arg1 rates 1009:1013 wild-type hyphal fusion rates 985:1013 wild-type hyphal fusion rates 985:1013 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 26254006 4 62 theme cell 770:773 arg1 glycans 780:786 cell wall glycans 770:786 cell wall glycans 770:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 7 63 theme gene 1213:1216 arg1 Deletion 1196:1203 Deletion 1196:1203 Deletion of gnt2 gene 1196:1216 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 6 64 theme reduced 1105:1111 arg1 levels 1113:1118 reduced levels 1105:1118 reduced levels of proteins involved in cell wall organization, biogenesis and remodelling 1105:1193 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 8 65 theme F. 1680:1681 arg1 oxysporum 1683:1691 F. oxysporum 1680:1691 F. oxysporum 1680:1691 Altogether, the results confirm that Gnt2-mediated N-glycosylation plays a crucial role in morphogenesis and virulence, and demonstrate that Gnt2 is essential for protein function, transport and relative abundance in F. oxysporum. 26254006 1 66 theme Protein 142:148 arg1 modification 201:212 a ubiquitous post-translational modification 169:212 a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization 169:303 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 1 66 theme Protein 142:148 arg1 N-glycosylation 150:164 Protein N-glycosylation 142:164 Protein N-glycosylation 142:164 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 0 67 from content 111:117 arg1 oxysporum 131:139 Fusarium oxysporum 122:139 Fusarium oxysporum 122:139 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 4 68 theme polymerization 840:853 arg1 degrees 855:861 polymerization degrees 840:861 polymerization degrees 840:861 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 0 69 theme wall 86:89 arg1 glycans 91:97 cell wall glycans 81:97 cell wall glycans 81:97 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 7 70 theme plant 1391:1395 arg1 walls 1402:1406 plant cell walls 1391:1406 plant cell walls 1391:1406 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 71 from size 736:739 arg1 glycans 780:786 cell wall glycans 770:786 cell wall glycans 770:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 1 72 theme ubiquitous 171:180 arg1 modification 201:212 a ubiquitous post-translational modification 169:212 a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization 169:303 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 1 72 theme ubiquitous 171:180 arg1 N-glycosylation 150:164 Protein N-glycosylation 142:164 Protein N-glycosylation 142:164 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 4 73 theme patterns 902:909 arg1 range 831:835 a broad range 823:835 a broad range of polymerization degrees and differential protein glycosylation patterns 823:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 0 74 theme Fusarium 122:129 arg1 oxysporum 131:139 Fusarium oxysporum 122:139 Fusarium oxysporum 122:139 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 1 75 theme protein 246:252 arg1 folding 254:260 appropriate protein folding 234:260 appropriate protein folding 234:260 Protein N-glycosylation is a ubiquitous post-translational modification that contributes to appropriate protein folding, stability, functionality and localization. 26254006 0 76 theme proteomic 12:20 arg1 analyses 22:29 Comparative proteomic analyses 0:29 Comparative proteomic analyses 0:29 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 6 77 gly Hypo-N-glycosylation 1016:1035 arg1 mutant 1046:1051 ∆gnt2 mutant 1040:1051 ∆gnt2 mutant 1040:1051 Hypo-N-glycosylation in ∆gnt2 mutant results in enhanced oxidative stress resistance and reduced levels of proteins involved in cell wall organization, biogenesis and remodelling. 26254006 4 78 theme protein 880:886 arg1 patterns 902:909 differential protein glycosylation patterns 867:909 differential protein glycosylation patterns 867:909 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 2 79 theme Fusarium 436:443 arg1 oxysporum 445:453 Fusarium oxysporum 436:453 Fusarium oxysporum 436:453 N-glycosylation has been identified as an important process for morphogenesis and virulence in several fungal pathogens including Fusarium oxysporum. 26254006 7 80 theme related 1322:1328 arg1 proteins 1313:1320 extracellular proteins 1299:1320 extracellular proteins related to detoxification of antifungal compounds 1299:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 7 81 theme extracellular 1299:1311 arg1 proteins 1313:1320 extracellular proteins 1299:1320 extracellular proteins related to detoxification of antifungal compounds 1299:1370 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 3 82 theme comparative 474:484 arg1 analyses 514:521 comparative chemical and proteome-based analyses 474:521 comparative chemical and proteome-based analyses 474:521 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 3 83 theme F. 618:619 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 3 84 theme N-glycosyltransferase 631:651 arg1 mutant 668:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 F. oxysporum N-glycosyltransferase Gnt2-deficient mutant 618:673 Here we conducted comparative chemical and proteome-based analyses to better understand the physiological changes associated with protein hypo-N-glycosylation in F. oxysporum N-glycosyltransferase Gnt2-deficient mutant. 26254006 0 85 gly N-glycosylation 57:71 arg1 cell 81:84 cell wall glycans 81:97 cell wall glycans 81:97 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 0 85 gly N-glycosylation 57:71 arg1 wall 86:89 cell wall glycans 81:97 cell wall glycans 81:97 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 0 85 gly N-glycosylation 57:71 arg1 protein 103:109 protein content 103:117 protein content in Fusarium oxysporum 103:139 Comparative proteomic analyses reveal that Gnt2-mediated N-glycosylation affects cell wall glycans and protein content in Fusarium oxysporum. 26254006 7 86 theme polygalacturonase 1435:1451 arg1 activity 1453:1460 extracellular polygalacturonase activity 1421:1460 extracellular polygalacturonase activity 1421:1460 Deletion of gnt2 gene led to accumulation of trafficking vesicles at hyphal tips, reduced secretion of extracellular proteins related to detoxification of antifungal compounds and degradation of plant cell walls, and lowered extracellular polygalacturonase activity. 26254006 4 87 theme galactofuranose 744:758 arg1 chains 760:765 galactofuranose chains 744:765 galactofuranose chains in cell wall glycans 744:786 The results suggest that lack of functional Gnt2 alters the size of galactofuranose chains in cell wall glycans, resulting in polysaccharides with a broad range of polymerization degrees and differential protein glycosylation patterns. 26254006 5 88 theme conidium 952:959 arg1 size 961:964 normal conidium size 945:964 normal conidium size 945:964 Functional Gnt2 is necessary for normal conidium size and morphology and wild-type hyphal fusion rates. 27199480 8 0 theme overall 1562:1568 arg1 fold 1570:1573 the overall fold 1558:1573 the overall fold 1558:1573 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 5 1 theme LPS 678:680 arg1 assembly 682:689 LPS assembly 678:689 LPS assembly 678:689 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 8 2 theme evolutionary 1497:1508 arg1 evident 1545:1551 evident 1545:1551 evident 1545:1551 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 8 2 theme evolutionary 1497:1508 arg1 connection 1510:1519 an evolutionary connection 1494:1519 an evolutionary connection that is not immediately evident from the overall fold or sequence similarities 1494:1598 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 3 3 theme core 328:331 arg1 oligosaccharide 333:347 the core oligosaccharide 324:347 the core oligosaccharide 324:347 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 8 4 theme sequence 1578:1585 arg1 similarities 1587:1598 sequence similarities 1578:1598 sequence similarities 1578:1598 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 6 5 theme recognized 873:882 arg1 families 904:911 the 98 currently recognized glycosyltransferase families 856:911 the 98 currently recognized glycosyltransferase families 856:911 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 7 6 from WbbB 1041:1044 arg1 function 1003:1010 function 1003:1010 function 1003:1010 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 6 from WbbB 1041:1044 arg1 structure 989:997 the crystallographic structure 968:997 the crystallographic structure 968:997 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 7 theme enzymes 1189:1195 arg1 typical 1173:1179 typical 1173:1179 typical 1173:1179 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 8 8 from site 1375:1378 arg1 domain 1402:1407 the C-terminal α/β domain 1383:1407 the C-terminal α/β domain 1383:1407 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 7 9 theme β-Kdo 1027:1031 arg1 GT 1033:1034 a prototype β-Kdo GT 1015:1034 a prototype β-Kdo GT 1015:1034 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 8 10 theme C-terminal 1387:1396 arg1 domain 1402:1407 the C-terminal α/β domain 1383:1407 the C-terminal α/β domain 1383:1407 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 5 11 dep structure 567:575 arg1 The 563:565 The 563:565 The 563:565 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 7 12 theme unique 1266:1271 arg1 architecture 1273:1284 a unique architecture 1264:1284 a unique architecture that makes it a prototype for a new GT family (GT99) 1264:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 13 theme GT-B 1184:1187 arg1 enzymes 1189:1195 GT-B enzymes 1184:1195 GT-B enzymes 1184:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 5 14 theme prototypical 597:608 arg1 glycosyltransferase 653:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 7 15 theme GT 1322:1323 arg1 GT99 1333:1336 GT99 1333:1336 GT99 1333:1336 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 15 theme GT 1322:1323 arg1 family 1325:1330 a new GT family 1316:1330 a new GT family (GT99) 1316:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 16 dep Raoultella 1109:1118 arg1 result 1254:1259 result 1254:1259 result in a unique architecture that makes it a prototype for a new GT family (GT99) 1254:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 16 dep Raoultella 1109:1118 arg1 terrigena 1120:1128 Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99) 1109:1337 Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99) 1109:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 16 dep Raoultella 1109:1118 arg1 has 1143:1145 has 1143:1145 has dual Rossmann-fold motifs typical of GT-B enzymes 1143:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 8 17 theme bacterial 1453:1461 arg1 sialyltransferases 1463:1480 bacterial sialyltransferases 1453:1480 bacterial sialyltransferases 1453:1480 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 5 18 theme monophosphate-Kdo-dependent 619:645 arg1 glycosyltransferase 653:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 5 19 theme cytidine 610:617 arg1 glycosyltransferase 653:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 6 20 theme β-Kdo 731:735 arg1 enzymes 793:799 glycosyltransferase enzymes 773:799 glycosyltransferase enzymes 773:799 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 6 20 theme β-Kdo 731:735 arg1 counterparts 737:748 the β-Kdo counterparts 727:748 the β-Kdo counterparts 727:748 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 1 21 theme Gram-negative 174:186 arg1 bacteria 188:195 Gram-negative bacteria 174:195 Gram-negative bacteria 174:195 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. 27199480 3 22 theme bacterial 356:364 arg1 LPSs 366:369 all bacterial LPSs 352:369 all bacterial LPSs 352:369 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 5 23 theme α-Kdo 647:651 arg1 glycosyltransferase 653:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase 593:671 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 8 24 theme α/β 1398:1400 arg1 domain 1402:1407 the C-terminal α/β domain 1383:1407 the C-terminal α/β domain 1383:1407 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 3 25 theme capsular 433:440 arg1 polysaccharides 442:456 group 2" capsular polysaccharides 424:456 "group 2" capsular polysaccharides 423:456 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 0 26 theme β-Kdo 10:14 arg1 glycosyltransferases 16:35 Bacterial β-Kdo glycosyltransferases 0:35 Bacterial β-Kdo glycosyltransferases 0:35 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 5 27 theme glycosyltransferase 653:671 arg1 function 581:588 function 581:588 function 581:588 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 5 27 theme glycosyltransferase 653:671 arg1 structure 567:575 structure 567:575 structure 567:575 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 7 28 theme prototype 1017:1025 arg1 GT 1033:1034 a prototype β-Kdo GT 1015:1034 a prototype β-Kdo GT 1015:1034 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 8 29 from site 1445:1448 arg1 sialyltransferases 1463:1480 bacterial sialyltransferases 1453:1480 bacterial sialyltransferases 1453:1480 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 3 30 theme group 424:428 arg1 polysaccharides 442:456 group 2" capsular polysaccharides 424:456 "group 2" capsular polysaccharides 423:456 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 0 31 theme Bacterial 0:8 arg1 glycosyltransferases 16:35 Bacterial β-Kdo glycosyltransferases 0:35 Bacterial β-Kdo glycosyltransferases 0:35 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 4 32 theme bacterial 536:544 arg1 polysaccharides 546:560 other bacterial polysaccharides 530:560 other bacterial polysaccharides 530:560 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 7 33 theme Rossmann-fold 1152:1164 arg1 motifs 1166:1171 dual Rossmann-fold motifs 1147:1171 dual Rossmann-fold motifs typical of GT-B enzymes 1147:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 0 34 theme glycosyltransferase 53:71 arg1 GT99 81:84 GT99 81:84 GT99 81:84 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 0 34 theme glycosyltransferase 53:71 arg1 family 73:78 a new glycosyltransferase family 47:78 a new glycosyltransferase family (GT99) 47:85 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 6 35 theme Carbohydrate-Active 920:938 arg1 database 948:955 the Carbohydrate-Active Enzymes database 916:955 the Carbohydrate-Active Enzymes database 916:955 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 8 36 from fold 1570:1573 arg1 evident 1545:1551 evident 1545:1551 evident 1545:1551 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 8 36 from fold 1570:1573 arg1 connection 1510:1519 an evolutionary connection 1494:1519 an evolutionary connection that is not immediately evident from the overall fold or sequence similarities 1494:1598 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 4 37 theme other 530:534 arg1 polysaccharides 546:560 other bacterial polysaccharides 530:560 other bacterial polysaccharides 530:560 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 7 38 theme GT 1033:1034 arg1 function 1003:1010 function 1003:1010 function 1003:1010 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 38 theme GT 1033:1034 arg1 structure 989:997 the crystallographic structure 968:997 the crystallographic structure 968:997 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 1 39 theme 3-deoxy-d-manno-oct-2-ulosonic 93:122 arg1 Kdo 88:90 Kdo 88:90 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) 88:128 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. 27199480 1 39 theme 3-deoxy-d-manno-oct-2-ulosonic 93:122 arg1 acid 124:127 3-deoxy-d-manno-oct-2-ulosonic acid 93:127 3-deoxy-d-manno-oct-2-ulosonic acid 93:127 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. 27199480 8 40 from similarities 1587:1598 arg1 evident 1545:1551 evident 1545:1551 evident 1545:1551 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 8 40 from similarities 1587:1598 arg1 connection 1510:1519 an evolutionary connection 1494:1519 an evolutionary connection that is not immediately evident from the overall fold or sequence similarities 1494:1598 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 6 41 theme Enzymes 940:946 arg1 database 948:955 the Carbohydrate-Active Enzymes database 916:955 the Carbohydrate-Active Enzymes database 916:955 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 7 42 contain has 1143:1145 arg1 GT 1140:1141 The β-Kdo GT 1130:1141 The β-Kdo GT 1130:1141 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 42 contain has 1143:1145 arg2 motifs 1166:1171 dual Rossmann-fold motifs 1147:1171 dual Rossmann-fold motifs typical of GT-B enzymes 1147:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 3 43 theme lyso 462:465 arg1 phosphatidylglycerol 467:486 (lyso)phosphatidylglycerol 461:486 (lyso)phosphatidylglycerol 461:486 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 0 44 theme new 49:51 arg1 GT99 81:84 GT99 81:84 GT99 81:84 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 0 44 theme new 49:51 arg1 family 73:78 a new glycosyltransferase family 47:78 a new glycosyltransferase family (GT99) 47:85 Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99). 27199480 4 45 theme polysaccharides 546:560 arg1 number 520:525 a small number 512:525 a small number of other bacterial polysaccharides 512:560 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 3 46 attach links 417:421 arg1 phosphatidylglycerol 467:486 (lyso)phosphatidylglycerol 461:486 (lyso)phosphatidylglycerol 461:486 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 3 46 attach links 417:421 arg2 oligosaccharide 383:397 an oligosaccharide 380:397 an oligosaccharide of β-Kdo residues 380:415 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 3 47 theme " 431:431 arg1 polysaccharides 442:456 group 2" capsular polysaccharides 424:456 "group 2" capsular polysaccharides 423:456 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 4 48 theme small 514:518 arg1 number 520:525 a small number 512:525 a small number of other bacterial polysaccharides 512:560 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 7 49 theme β-Kdo 1134:1138 arg1 GT 1140:1141 The β-Kdo GT 1130:1141 The β-Kdo GT 1130:1141 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 6 50 theme bioinformatics 804:817 arg1 tools 819:823 bioinformatics tools 804:823 bioinformatics tools 804:823 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 3 51 theme β-Kdo 402:406 arg1 residues 408:415 β-Kdo residues 402:415 β-Kdo residues 402:415 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 7 52 theme dual 1147:1150 arg1 motifs 1166:1171 dual Rossmann-fold motifs 1147:1171 dual Rossmann-fold motifs typical of GT-B enzymes 1147:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 2 53 theme surface 232:238 arg1 polysaccharides 240:254 surface polysaccharides 232:254 surface polysaccharides 232:254 It is often involved in attaching surface polysaccharides to their lipid anchors. 27199480 7 54 theme O-antigen 1086:1094 arg1 synthesis 1096:1104 LPS O-antigen synthesis 1082:1104 LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99) 1082:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 3 55 theme residues 408:415 arg1 oligosaccharide 383:397 an oligosaccharide 380:397 an oligosaccharide of β-Kdo residues 380:415 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 7 56 theme typical 1173:1179 arg1 motifs 1166:1171 dual Rossmann-fold motifs 1147:1171 dual Rossmann-fold motifs typical of GT-B enzymes 1147:1195 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 5 57 from assembly 682:689 arg1 function 581:588 function 581:588 function 581:588 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 5 57 from assembly 682:689 arg1 structure 567:575 structure 567:575 structure 567:575 The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. 27199480 6 58 theme glycosyltransferase 884:902 arg1 families 904:911 the 98 currently recognized glycosyltransferase families 856:911 the 98 currently recognized glycosyltransferase families 856:911 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 7 59 theme extensive 1202:1210 arg1 deletions 1212:1220 extensive deletions 1202:1220 extensive deletions 1202:1220 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 6 60 theme glycosyltransferase 773:791 arg1 enzymes 793:799 glycosyltransferase enzymes 773:799 glycosyltransferase enzymes 773:799 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 6 60 theme glycosyltransferase 773:791 arg1 counterparts 737:748 the β-Kdo counterparts 727:748 the β-Kdo counterparts 727:748 In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. 27199480 4 61 located found 503:507 arg1 number 520:525 a small number 512:525 a small number of other bacterial polysaccharides 512:560 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 4 61 located found 503:507 arg2 β-Kdo 489:493 β-Kdo 489:493 β-Kdo 489:493 β-Kdo is also found in a small number of other bacterial polysaccharides. 27199480 8 62 theme corresponding 1431:1443 arg1 site 1445:1448 the corresponding site 1427:1448 the corresponding site in bacterial sialyltransferases 1427:1480 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 7 63 from synthesis 1096:1104 arg1 Raoultella 1109:1118 Raoultella 1109:1118 Raoultella 1109:1118 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 3 64 theme lipid 312:316 arg1 A 318:318 lipid A 312:318 lipid A 312:318 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 3 65 from oligosaccharide 333:347 arg1 LPSs 366:369 all bacterial LPSs 352:369 all bacterial LPSs 352:369 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 7 66 theme LPS 1082:1084 arg1 synthesis 1096:1104 LPS O-antigen synthesis 1082:1104 LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99) 1082:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 67 theme modular 1049:1055 arg1 protein 1057:1063 a modular protein 1047:1063 a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99) 1047:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 67 theme modular 1049:1055 arg1 WbbB 1041:1044 WbbB 1041:1044 WbbB 1041:1044 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 1 68 theme eight-carbon 136:147 arg1 Kdo 88:90 Kdo 88:90 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) 88:128 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. 27199480 1 68 theme eight-carbon 136:147 arg1 sugar 149:153 an eight-carbon sugar 133:153 an eight-carbon sugar mostly confined to Gram-negative bacteria 133:195 Kdo (3-deoxy-d-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. 27199480 7 69 theme new 1318:1320 arg1 GT99 1333:1336 GT99 1333:1336 GT99 1333:1336 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 7 69 theme new 1318:1320 arg1 family 1325:1330 a new GT family 1316:1330 a new GT family (GT99) 1316:1337 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 2 70 theme lipid 265:269 arg1 anchors 271:277 their lipid anchors 259:277 their lipid anchors 259:277 It is often involved in attaching surface polysaccharides to their lipid anchors. 27199480 8 71 theme monophosphate-binding 1353:1373 arg1 site 1375:1378 The cytidine monophosphate-binding site 1340:1378 The cytidine monophosphate-binding site in the C-terminal α/β domain 1340:1407 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 27199480 7 72 theme crystallographic 972:987 arg1 structure 989:997 the crystallographic structure 968:997 the crystallographic structure 968:997 We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). 27199480 3 73 from A 318:318 arg1 LPSs 366:369 all bacterial LPSs 352:369 all bacterial LPSs 352:369 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 3 74 dep provides 286:293 arg1 whereas 372:378 whereas 372:378 whereas 372:378 α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. 27199480 8 75 theme cytidine 1344:1351 arg1 site 1375:1378 The cytidine monophosphate-binding site 1340:1378 The cytidine monophosphate-binding site in the C-terminal α/β domain 1340:1407 The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. 24702330 3 0 theme encoded 371:377 arg1 tag 388:390 a genetically encoded aldehyde tag 357:390 a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 357:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 4 1 theme amino-acid 534:543 arg1 sequence 545:552 a five amino-acid sequence 527:552 a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration 527:712 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 6 2 theme protein 1026:1032 arg1 machineries 1048:1058 natural protein glycosylation machineries 1018:1058 natural protein glycosylation machineries 1018:1058 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 3 3 theme aldehyde 379:386 arg1 tag 388:390 a genetically encoded aldehyde tag 357:390 a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 357:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 5 4 theme glycan 895:900 arg1 oxazolines 902:911 corresponding glycan oxazolines 881:911 corresponding glycan oxazolines 881:911 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 4 5 theme Fc 496:497 arg1 sequon 515:520 the natural Fc N-glycosylation sequon 484:520 the natural Fc N-glycosylation sequon 484:520 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 6 6 theme Fc 985:986 arg1 glycoforms 988:997 specific Fc glycoforms 976:997 specific Fc glycoforms 976:997 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 3 7 theme glycosylation 408:420 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 7 theme glycosylation 408:420 arg1 site 422:425 the internal glycosylation site 395:425 the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 395:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 5 8 from oxazolines 902:911 arg1 transfer 846:853 enzymatic transfer 836:853 enzymatic transfer of complex N-glycans from corresponding glycan oxazolines 836:911 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 1 9 gly glycosylation 100:112 arg2 sites 114:118 chemically defined glycosylation sites 81:118 chemically defined glycosylation sites 81:118 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 5 10 theme modified 812:819 arg1 Fc 821:822 the modified Fc 808:822 the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase 808:945 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 0 11 theme Chemoenzymatic 0:13 arg1 glycosylation 18:30 Chemoenzymatic Fc glycosylation 0:30 Chemoenzymatic Fc glycosylation via engineered aldehyde tags 0:59 Chemoenzymatic Fc glycosylation via engineered aldehyde tags. 24702330 6 12 theme natural 1018:1024 arg1 machineries 1048:1058 natural protein glycosylation machineries 1018:1058 natural protein glycosylation machineries 1018:1058 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 5 13 theme EndoS-derived 919:931 arg1 glycosynthase 933:945 an EndoS-derived glycosynthase 916:945 an EndoS-derived glycosynthase 916:945 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 3 14 theme internal 399:406 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 14 theme internal 399:406 arg1 site 422:425 the internal glycosylation site 395:425 the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 395:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 0 15 theme aldehyde 47:54 arg1 tags 56:59 engineered aldehyde tags 36:59 engineered aldehyde tags 36:59 Chemoenzymatic Fc glycosylation via engineered aldehyde tags. 24702330 6 16 theme specific 976:983 arg1 glycoforms 988:997 specific Fc glycoforms 976:997 specific Fc glycoforms 976:997 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 1 17 theme defined 92:98 arg1 sites 114:118 chemically defined glycosylation sites 81:118 chemically defined glycosylation sites 81:118 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 5 18 link Oxime-linked 715:726 arg1 glycoconjugates 728:742 Oxime-linked glycoconjugates 715:742 Oxime-linked glycoconjugates 715:742 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 0 19 theme Fc 15:16 arg1 glycosylation 18:30 Chemoenzymatic Fc glycosylation 0:30 Chemoenzymatic Fc glycosylation via engineered aldehyde tags 0:59 Chemoenzymatic Fc glycosylation via engineered aldehyde tags. 24702330 5 20 theme corresponding 881:893 arg1 oxazolines 902:911 corresponding glycan oxazolines 881:911 corresponding glycan oxazolines 881:911 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 5 21 theme complex 858:864 arg1 N-glycans 866:874 complex N-glycans 858:874 complex N-glycans 858:874 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 1 22 theme glycosylation 100:112 arg1 sites 114:118 chemically defined glycosylation sites 81:118 chemically defined glycosylation sites 81:118 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 3 23 theme IgG1 466:469 arg1 Fc 459:460 Fc 459:460 Fc 459:460 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 23 theme IgG1 466:469 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 24 theme fragment 449:456 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 24 theme fragment 449:456 arg1 site 422:425 the internal glycosylation site 395:425 the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 395:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 4 25 theme generating 614:623 arg1 enzyme 625:630 recombinant formylglycine generating enzyme 588:630 recombinant formylglycine generating enzyme 588:630 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 5 26 theme Oxime-linked 715:726 arg1 glycoconjugates 728:742 Oxime-linked glycoconjugates 715:742 Oxime-linked glycoconjugates 715:742 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 6 27 gly glycoforms 988:997 arg1 Fc 985:986 specific Fc glycoforms 976:997 specific Fc glycoforms 976:997 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 4 28 theme formylglycine 600:612 arg1 enzyme 625:630 recombinant formylglycine generating enzyme 588:630 recombinant formylglycine generating enzyme 588:630 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 3 29 gly glycosylation 408:420 arg2 site 422:425 the internal glycosylation site 395:425 the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 395:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 29 gly glycosylation 408:420 arg1 Fc 459:460 Fc 459:460 Fc 459:460 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 29 gly glycosylation 408:420 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 29 gly glycosylation 408:420 arg2 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 4 30 theme natural 488:494 arg1 sequon 515:520 the natural Fc N-glycosylation sequon 484:520 the natural Fc N-glycosylation sequon 484:520 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 4 31 theme chemical 693:700 arg1 elaboration 702:712 subsequent chemical elaboration 682:712 subsequent chemical elaboration 682:712 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 2 32 gly glycosylation 278:290 arg1 proteins 311:318 aldehyde-tagged proteins 295:318 aldehyde-tagged proteins 295:318 One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. 24702330 5 33 link EndoS-derived 919:931 arg1 glycosynthase 933:945 an EndoS-derived glycosynthase 916:945 an EndoS-derived glycosynthase 916:945 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 4 34 gly N-glycosylation 499:513 arg2 sequon 515:520 the natural Fc N-glycosylation sequon 484:520 the natural Fc N-glycosylation sequon 484:520 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 0 35 theme engineered 36:45 arg1 tags 56:59 engineered aldehyde tags 36:59 engineered aldehyde tags 36:59 Chemoenzymatic Fc glycosylation via engineered aldehyde tags. 24702330 4 36 theme N-glycosylation 499:513 arg1 sequon 515:520 the natural Fc N-glycosylation sequon 484:520 the natural Fc N-glycosylation sequon 484:520 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 1 37 with Glycoproteins 62:74 arg1 sites 114:118 chemically defined glycosylation sites 81:118 chemically defined glycosylation sites 81:118 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 1 37 with Glycoproteins 62:74 arg1 structures 124:133 structures 124:133 structures 124:133 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 3 38 from site 422:425 arg1 tag 388:390 a genetically encoded aldehyde tag 357:390 a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 357:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 38 from site 422:425 arg1 installation 341:352 the installation 337:352 the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 337:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 2 39 theme chemical 269:276 arg1 glycosylation 278:290 chemical glycosylation 269:290 chemical glycosylation of aldehyde-tagged proteins 269:318 One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. 24702330 6 40 theme glycosylation 1034:1046 arg1 machineries 1048:1058 natural protein glycosylation machineries 1018:1058 natural protein glycosylation machineries 1018:1058 In this manner we generated specific Fc glycoforms without relying on natural protein glycosylation machineries. 24702330 4 41 theme recombinant 588:598 arg1 enzyme 625:630 recombinant formylglycine generating enzyme 588:630 recombinant formylglycine generating enzyme 588:630 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 5 42 theme aminooxy 776:783 arg1 N-acetylglucosamine 785:803 aminooxy N-acetylglucosamine 776:803 aminooxy N-acetylglucosamine 776:803 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 1 43 theme important 139:147 arg1 Glycoproteins 62:74 Glycoproteins 62:74 Glycoproteins with chemically defined glycosylation sites and structures 62:133 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 1 43 theme important 139:147 arg1 targets 167:173 important biopharmaceutical targets 139:173 important biopharmaceutical targets 139:173 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 2 44 theme such 245:248 arg1 molecules 250:258 such molecules 245:258 such molecules 245:258 One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. 24702330 1 45 theme biopharmaceutical 149:165 arg1 Glycoproteins 62:74 Glycoproteins 62:74 Glycoproteins with chemically defined glycosylation sites and structures 62:133 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 1 45 theme biopharmaceutical 149:165 arg1 targets 167:173 important biopharmaceutical targets 139:173 important biopharmaceutical targets 139:173 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 3 46 from installation 341:352 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 46 from installation 341:352 arg1 site 422:425 the internal glycosylation site 395:425 the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 395:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 4 47 theme aldehyde 662:669 arg1 groups 671:676 aldehyde groups 662:676 aldehyde groups 662:676 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 3 48 theme tag 388:390 arg1 installation 341:352 the installation 337:352 the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1 337:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 2 49 theme proteins 311:318 arg1 glycosylation 278:290 chemical glycosylation 269:290 chemical glycosylation of aldehyde-tagged proteins 269:318 One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. 24702330 3 50 theme crystallizable 434:447 arg1 Fc 459:460 Fc 459:460 Fc 459:460 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 3 50 theme crystallizable 434:447 arg1 fragment 449:456 the crystallizable fragment 430:456 the crystallizable fragment (Fc) of IgG1 430:469 Here, we report the installation of a genetically encoded aldehyde tag at the internal glycosylation site of the crystallizable fragment (Fc) of IgG1. 24702330 4 51 theme subsequent 682:691 arg1 elaboration 702:712 subsequent chemical elaboration 682:712 subsequent chemical elaboration 682:712 We replaced the natural Fc N-glycosylation sequon with a five amino-acid sequence that was efficiently converted by recombinant formylglycine generating enzyme in vitro, thereby introducing aldehyde groups for subsequent chemical elaboration. 24702330 5 52 theme N-glycans 866:874 arg1 transfer 846:853 enzymatic transfer 836:853 enzymatic transfer of complex N-glycans from corresponding glycan oxazolines 836:911 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24702330 2 53 theme aldehyde-tagged 295:309 arg1 proteins 311:318 aldehyde-tagged proteins 295:318 aldehyde-tagged proteins 295:318 One approach toward constructing such molecules involves chemical glycosylation of aldehyde-tagged proteins. 24702330 1 54 theme critical 179:186 arg1 tools 188:192 critical tools 179:192 critical tools for glycobiology 179:209 Glycoproteins with chemically defined glycosylation sites and structures are important biopharmaceutical targets and critical tools for glycobiology. 24702330 5 55 theme enzymatic 836:844 arg1 transfer 846:853 enzymatic transfer 836:853 enzymatic transfer of complex N-glycans from corresponding glycan oxazolines 836:911 Oxime-linked glycoconjugates were synthesized by conjugating aminooxy N-acetylglucosamine to the modified Fc followed by enzymatic transfer of complex N-glycans from corresponding glycan oxazolines by an EndoS-derived glycosynthase. 24234447 6 0 theme confirmed 1099:1107 arg1 structures 1116:1125 experimentally confirmed glycan structures 1084:1125 experimentally confirmed glycan structures from the literature 1084:1145 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 2 1 theme processes 390:398 arg1 understanding 294:306 our understanding 290:306 our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes 290:398 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 1 2 dep UniCarbKB 89:97 arg1 http 100:103 http 100:103 http://unicarbkb.org 100:119 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 4 3 theme global 855:860 arg1 information 891:901 global and site-specific attachment information 855:901 information 891:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 8 4 theme previous 1341:1348 arg1 issues 1363:1368 previous NAR Database issues 1341:1368 previous NAR Database issues 1341:1368 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 3 5 theme high-quality 646:657 arg1 resource 671:678 a high-quality and updated resource 644:678 a high-quality and updated resource to support glycomics and glycoproteomics research 644:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 3 6 theme glycan 528:533 arg1 database 545:552 the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research 524:728 the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research 524:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 7 7 gly glycoproteins 1167:1179 arg1 glycoproteins 1167:1179 35 glycoproteins 1164:1179 35 glycoproteins 1164:1179 Among these are 35 glycoproteins, 502 structures and 60 publications previously not included in GlycoSuiteDB. 24234447 1 8 gly glycoproteins 217:229 arg1 glycoproteins 217:229 glycoproteins 217:229 glycoproteins 217:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 4 9 theme glycoprotein 820:831 arg1 information 833:843 published glycoprotein information 810:843 published glycoprotein information including global and site-specific attachment information 810:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 6 10 theme glycan 1109:1114 arg1 structures 1116:1125 experimentally confirmed glycan structures 1084:1125 experimentally confirmed glycan structures from the literature 1084:1145 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 2 11 theme structural 415:424 arg1 information 468:478 structural, experimental and functional glycoscience information 415:478 structural, experimental and functional glycoscience information 415:478 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 1 12 theme UniCarb 66:72 arg1 KnowledgeBase 74:86 The UniCarb KnowledgeBase 62:86 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) 62:120 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 9 13 theme data 1544:1547 arg1 collections 1549:1559 substantial new curated data collections 1520:1559 substantial new curated data collections 1520:1559 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 1 14 theme growing 148:154 arg1 database 165:172 a growing, curated database 146:172 a growing, curated database of information on the glycan structures of glycoproteins 146:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 9 15 theme substantial 1520:1530 arg1 collections 1549:1559 substantial new curated data collections 1520:1559 substantial new curated data collections 1520:1559 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 2 16 theme international 248:260 arg1 UniCarbKB 232:240 UniCarbKB 232:240 UniCarbKB 232:240 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 2 16 theme international 248:260 arg1 effort 262:267 an international effort 245:267 an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information 245:478 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 2 17 theme glyco-mediated 375:388 arg1 processes 390:398 glyco-mediated processes 375:398 glyco-mediated processes 375:398 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 2 18 theme glycoscience 455:466 arg1 information 468:478 structural, experimental and functional glycoscience information 415:478 structural, experimental and functional glycoscience information 415:478 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 6 19 theme glycosylation 1039:1051 arg1 sites 1053:1057 Further, 598 protein glycosylation sites 1018:1057 sites 1053:1057 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 6 20 from literature 1136:1145 arg1 structures 1116:1125 experimentally confirmed glycan structures 1084:1125 experimentally confirmed glycan structures from the literature 1084:1145 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 3 21 theme informatic 586:595 arg1 standards 597:605 the informatic standards 582:605 the informatic standards introduced by EUROCarbDB 582:630 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 5 22 theme glycan 952:957 arg1 entries 969:975 3740 glycan structure entries 947:975 3740 glycan structure entries 947:975 For the first release over 890 references, 3740 glycan structure entries and 400 glycoproteins have been curated. 24234447 2 23 theme functional 444:453 arg1 information 468:478 structural, experimental and functional glycoscience information 415:478 structural, experimental and functional glycoscience information 415:478 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 8 24 theme NAR 1350:1352 arg1 issues 1363:1368 previous NAR Database issues 1341:1368 previous NAR Database issues 1341:1368 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 6 25 gly glycosylation 1039:1051 arg2 sites 1053:1057 Further, 598 protein glycosylation sites 1018:1057 sites 1053:1057 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 6 25 gly glycosylation 1039:1051 arg2 598 1027:1029 598 1027:1029 598 1027:1029 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 8 26 with integration 1424:1434 arg1 GlycoMod 1455:1462 GlycoMod 1455:1462 GlycoMod 1455:1462 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 8 26 with integration 1424:1434 arg1 UniProtKB 1441:1449 UniProtKB 1441:1449 UniProtKB 1441:1449 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 5 27 theme structure 959:967 arg1 entries 969:975 3740 glycan structure entries 947:975 3740 glycan structure entries 947:975 For the first release over 890 references, 3740 glycan structure entries and 400 glycoproteins have been curated. 24234447 9 28 theme new 1532:1534 arg1 collections 1549:1559 substantial new curated data collections 1520:1559 substantial new curated data collections 1520:1559 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 4 29 theme published 810:818 arg1 information 833:843 published glycoprotein information 810:843 published glycoprotein information including global and site-specific attachment information 810:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 9 30 theme intuitive 1565:1573 arg1 user-interfaces 1575:1589 intuitive user-interfaces 1565:1589 intuitive user-interfaces that improve database searching 1565:1621 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 2 31 theme experimental 427:438 arg1 information 468:478 structural, experimental and functional glycoscience information 415:478 structural, experimental and functional glycoscience information 415:478 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 3 32 theme database 545:552 arg1 success 513:519 the success 509:519 the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research 509:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 1 33 theme information 177:187 arg1 database 165:172 a growing, curated database 146:172 a growing, curated database of information on the glycan structures of glycoproteins 146:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 4 34 theme comprehensive 750:762 arg1 information 764:774 comprehensive information 750:774 comprehensive information concerning glycan structures 750:803 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 9 35 theme database 1604:1611 arg1 searching 1613:1621 database searching 1604:1621 database searching 1604:1621 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 8 36 theme Database 1354:1361 arg1 issues 1363:1368 previous NAR Database issues 1341:1368 previous NAR Database issues 1341:1368 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 0 37 theme knowledge 22:30 arg1 platform 32:39 a knowledge platform 20:39 a knowledge platform for glycoproteomics 20:59 UniCarbKB: building a knowledge platform for glycoproteomics. 24234447 6 38 theme protein 1031:1037 arg1 sites 1053:1057 Further, 598 protein glycosylation sites 1018:1057 sites 1053:1057 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 8 39 theme GlycoSuiteDB 1315:1326 arg1 transformation 1297:1310 the transformation 1293:1310 the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod 1293:1462 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 9 40 theme curated 1536:1542 arg1 collections 1549:1559 substantial new curated data collections 1520:1559 substantial new curated data collections 1520:1559 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 5 41 theme first 912:916 arg1 release 918:924 the first release 908:924 the first release over 890 references 908:944 For the first release over 890 references, 3740 glycan structure entries and 400 glycoproteins have been curated. 24234447 3 42 theme updated 663:669 arg1 resource 671:678 a high-quality and updated resource 644:678 a high-quality and updated resource to support glycomics and glycoproteomics research 644:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 1 43 dep growing 148:154 arg1 curated 157:163 curated 157:163 curated 157:163 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 7 44 dep included 1232:1239 arg1 glycoproteins 1167:1179 35 glycoproteins 1164:1179 35 glycoproteins 1164:1179 Among these are 35 glycoproteins, 502 structures and 60 publications previously not included in GlycoSuiteDB. 24234447 7 44 dep included 1232:1239 arg1 structures 1186:1195 502 structures 1182:1195 502 structures 1182:1195 Among these are 35 glycoproteins, 502 structures and 60 publications previously not included in GlycoSuiteDB. 24234447 7 44 dep included 1232:1239 arg1 publications 1204:1215 60 publications 1201:1215 60 publications 1201:1215 Among these are 35 glycoproteins, 502 structures and 60 publications previously not included in GlycoSuiteDB. 24234447 2 45 theme pathways 323:330 arg1 understanding 294:306 our understanding 290:306 our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes 290:398 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 8 46 with UniCarbKB 1406:1414 arg1 GlycoMod 1455:1462 GlycoMod 1455:1462 GlycoMod 1455:1462 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 8 46 with UniCarbKB 1406:1414 arg1 UniProtKB 1441:1449 UniProtKB 1441:1449 UniProtKB 1441:1449 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 8 47 dep transformation 1297:1310 arg1 hosted 1374:1379 hosted 1374:1379 hosted by ExPASy since 2009 1374:1400 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 8 47 dep transformation 1297:1310 arg1 featured 1329:1336 featured 1329:1336 featured in previous NAR Database issues 1329:1368 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 8 48 from update 1283:1288 arg1 transformation 1297:1310 the transformation 1293:1310 the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod 1293:1462 This article provides an update on the transformation of GlycoSuiteDB (featured in previous NAR Database issues and hosted by ExPASy since 2009) to UniCarbKB and its integration with UniProtKB and GlycoMod. 24234447 1 49 dep KnowledgeBase 74:86 arg1 UniCarbKB 89:97 UniCarbKB 89:97 UniCarbKB 89:97 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 6 50 theme Further 1018:1024 arg1 sites 1053:1057 Further, 598 protein glycosylation sites 1018:1057 sites 1053:1057 Further, 598 protein glycosylation sites have been annotated with experimentally confirmed glycan structures from the literature. 24234447 5 51 gly glycoproteins 985:997 arg1 glycoproteins 985:997 400 glycoproteins 981:997 400 glycoproteins 981:997 For the first release over 890 references, 3740 glycan structure entries and 400 glycoproteins have been curated. 24234447 3 52 theme glycoproteomics 705:719 arg1 research 721:728 glycomics and glycoproteomics research 691:728 glycomics and glycoproteomics research 691:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 4 53 theme site-specific 866:878 arg1 information 891:901 global and site-specific attachment information 855:901 information 891:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 1 54 from information 177:187 arg1 structures 203:212 the glycan structures 192:212 the glycan structures of glycoproteins 192:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 2 55 theme structures 311:320 arg1 understanding 294:306 our understanding 290:306 our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes 290:398 UniCarbKB is an international effort that aims to further our understanding of structures, pathways and networks involved in glycosylation and glyco-mediated processes by integrating structural, experimental and functional glycoscience information. 24234447 1 56 theme glycan 196:201 arg1 structures 203:212 the glycan structures 192:212 the glycan structures of glycoproteins 192:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 1 57 from structures 203:212 arg1 database 165:172 a growing, curated database 146:172 a growing, curated database of information on the glycan structures of glycoproteins 146:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 0 58 dep building 11:18 arg1 UniCarbKB 0:8 UniCarbKB 0:8 UniCarbKB 0:8 UniCarbKB: building a knowledge platform for glycoproteomics. 24234447 3 59 theme structure 535:543 arg1 database 545:552 the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research 524:728 the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research 524:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 4 60 theme glycan 787:792 arg1 structures 794:803 glycan structures 787:803 glycan structures 787:803 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 4 61 gly glycoprotein 820:831 arg1 glycoprotein 820:831 published glycoprotein information 810:843 published glycoprotein information including global and site-specific attachment information 810:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 3 62 theme glycomics 691:699 arg1 research 721:728 glycomics and glycoproteomics research 691:728 glycomics and glycoproteomics research 691:728 This initiative builds upon the success of the glycan structure database GlycoSuiteDB, together with the informatic standards introduced by EUROCarbDB, to provide a high-quality and updated resource to support glycomics and glycoproteomics research. 24234447 1 63 from database 165:172 arg1 structures 203:212 the glycan structures 192:212 the glycan structures of glycoproteins 192:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 1 64 theme public 129:134 arg1 access 136:141 public access 129:141 public access to a growing, curated database of information on the glycan structures of glycoproteins 129:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 5 65 theme 3740 947:950 arg1 entries 969:975 3740 glycan structure entries 947:975 3740 glycan structure entries 947:975 For the first release over 890 references, 3740 glycan structure entries and 400 glycoproteins have been curated. 24234447 9 66 theme refactored 1486:1495 arg1 database 1497:1504 a refactored database 1484:1504 a refactored database 1484:1504 Here, we introduce a refactored database, supported by substantial new curated data collections and intuitive user-interfaces that improve database searching. 24234447 4 67 theme attachment 880:889 arg1 information 891:901 global and site-specific attachment information 855:901 information 891:901 UniCarbKB provides comprehensive information concerning glycan structures, and published glycoprotein information including global and site-specific attachment information. 24234447 1 68 dep http 100:103 arg1 //unicarbkb.org 105:119 //unicarbkb.org 105:119 http://unicarbkb.org 100:119 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 24234447 1 69 theme glycoproteins 217:229 arg1 structures 203:212 the glycan structures 192:212 the glycan structures of glycoproteins 192:229 The UniCarb KnowledgeBase (UniCarbKB; http://unicarbkb.org) offers public access to a growing, curated database of information on the glycan structures of glycoproteins. 27313494 7 0 from fractions 1362:1370 arg1 present 1342:1348 present 1342:1348 present 1342:1348 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 6 1 from Jacalin 1036:1042 arg1 KK47 1065:1068 KK47 1065:1068 KK47 1065:1068 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 3 2 from Diversity 310:318 arg1 cells 352:356 animal cells 345:356 animal cells 345:356 Diversity of glycan composition in animal cells remains poorly understood. 27313494 4 3 theme subcellular 448:458 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 4 theme mitochondrial 643:655 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 7 5 from N-glycans 1380:1388 arg1 YTS1 1393:1396 YTS1 1393:1396 YTS1 1393:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 5 from N-glycans 1380:1388 arg1 HCV29 1331:1335 HCV29 1331:1335 HCV29 1331:1335 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 6 6 from cell 1025:1028 arg1 KK47 1065:1068 KK47 1065:1068 KK47 1065:1068 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 0 7 theme Global 0:5 arg1 Identification 7:20 Global Identification 0:20 Global Identification 0:20 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 6 8 theme significant 899:909 arg1 differences 911:921 significant differences 899:921 significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell 899:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 9 9 theme functional 1759:1768 arg1 roles 1770:1774 the functional roles 1755:1774 the functional roles of glycans and corresponding glycosylated proteins in distinct organelles 1755:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 6 10 from STL 1031:1033 arg1 KK47 1065:1068 KK47 1065:1068 KK47 1065:1068 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 1 11 theme cellular 216:223 arg1 function 225:232 cellular function 216:232 cellular function 216:232 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 7 12 from N-glycans 1318:1326 arg1 YTS1 1393:1396 YTS1 1393:1396 YTS1 1393:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 12 from N-glycans 1318:1326 arg1 HCV29 1331:1335 HCV29 1331:1335 HCV29 1331:1335 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 13 from fractions 1162:1170 arg1 total 1121:1125 a total 1119:1125 a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis 1119:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 4 14 dep mitochondrial 643:655 arg1 Mito 658:661 Mito 658:661 Mito 658:661 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 15 theme YTS1 516:519 arg1 cells 521:525 metastatic bladder YTS1 cells 497:525 metastatic bladder YTS1 cells 497:525 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 7 16 theme MS 1199:1200 arg1 analysis 1202:1209 MS analysis 1199:1209 MS analysis 1199:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 5 17 theme lectin 735:740 arg1 microarray 742:751 lectin microarray 735:751 lectin microarray 735:751 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 7 18 from present 1342:1348 arg1 fractions 1362:1370 all four fractions 1353:1370 all four fractions 1353:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 6 19 theme binding 967:973 arg1 terms 951:955 terms 951:955 terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell 951:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 3 20 theme animal 345:350 arg1 cells 352:356 animal cells 345:356 animal cells 345:356 Diversity of glycan composition in animal cells remains poorly understood. 27313494 9 21 from roles 1770:1774 arg1 organelles 1839:1848 distinct organelles 1830:1848 distinct organelles 1830:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 9 22 theme glycosylated 1805:1816 arg1 proteins 1818:1825 corresponding glycosylated proteins 1791:1825 corresponding glycosylated proteins 1791:1825 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 1 23 theme cellular 139:146 arg1 components 148:157 cellular components 139:157 cellular components 139:157 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 7 24 from N-glycans 1273:1281 arg1 YTS1 1286:1289 YTS1 1286:1289 YTS1 1286:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 24 from N-glycans 1273:1281 arg1 N-glycans 1318:1326 KK47 and 7 N-glycans 1307:1326 KK47 and 7 N-glycans in HCV29 were present in all four fractions 1307:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 24 from N-glycans 1273:1281 arg1 HCV29 1440:1444 HCV29 1440:1444 HCV29 1440:1444 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 24 from N-glycans 1273:1281 arg1 N-glycans 1380:1388 10 N-glycans 1377:1388 10 N-glycans in YTS1 1377:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 24 from N-glycans 1273:1281 arg1 KK47 1415:1418 KK47 1415:1418 KK47 1415:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 3 25 theme composition 330:340 arg1 Diversity 310:318 Diversity 310:318 Diversity of glycan composition in animal cells 310:356 Diversity of glycan composition in animal cells remains poorly understood. 27313494 7 26 from total 1121:1125 arg1 fractions 1162:1170 four fractions 1157:1170 four fractions of three cells detected by MS analysis 1157:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 27 theme fucosylated 1229:1239 arg1 structures 1241:1250 fucosylated structures 1229:1250 fucosylated structures 1229:1250 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 4 28 from homogenate 483:492 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 2 29 theme protein 291:297 arg1 functions 299:307 protein functions 291:307 protein functions 291:307 Protein glycosylation provides a basis for diversity of protein functions. 27313494 4 30 theme metastatic 497:506 arg1 cells 521:525 metastatic bladder YTS1 cells 497:525 metastatic bladder YTS1 cells 497:525 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 31 from elucidation 1740:1750 arg1 organelles 1839:1848 distinct organelles 1830:1848 distinct organelles 1830:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 7 32 located present 1342:1348 arg1 fractions 1362:1370 all four fractions 1353:1370 all four fractions 1353:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 32 located present 1342:1348 arg2 N-glycans 1318:1326 KK47 and 7 N-glycans 1307:1326 KK47 and 7 N-glycans in HCV29 were present in all four fractions 1307:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 9 33 theme integrated 1583:1592 arg1 strategy 1594:1601 The integrated strategy 1579:1601 The integrated strategy described here 1579:1616 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 9 33 theme integrated 1583:1592 arg1 useful 1729:1734 useful 1729:1734 useful 1729:1734 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 34 theme cancer 565:570 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 7 35 from N-glycans 1427:1435 arg1 YTS1 1286:1289 YTS1 1286:1289 YTS1 1286:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 35 from N-glycans 1427:1435 arg1 N-glycans 1318:1326 KK47 and 7 N-glycans 1307:1326 KK47 and 7 N-glycans in HCV29 were present in all four fractions 1307:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 35 from N-glycans 1427:1435 arg1 HCV29 1440:1444 HCV29 1440:1444 HCV29 1440:1444 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 35 from N-glycans 1427:1435 arg1 N-glycans 1380:1388 10 N-glycans 1377:1388 10 N-glycans in YTS1 1377:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 35 from N-glycans 1427:1435 arg1 KK47 1415:1418 KK47 1415:1418 KK47 1415:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 1 36 theme processes 191:199 arg1 Compartmentalization 115:134 Compartmentalization 115:134 Compartmentalization of cellular components and their associated biological processes 115:199 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 4 37 theme cells 577:581 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 8 38 from Glycans 1481:1487 arg1 category 1503:1510 the latter category 1492:1510 the latter category 1492:1510 Glycans in the latter category are considered potential markers for the corresponding organelles. 27313494 0 39 theme Bladder 100:106 arg1 Cells 108:112 Bladder Cells 100:112 Bladder Cells 100:112 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 4 40 theme grade 532:536 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 41 from VVA 1086:1088 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 41 from VVA 1086:1088 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 41 from VVA 1086:1088 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 5 42 theme spectrometry 762:773 arg1 analysis 780:787 mass spectrometry (MS) analysis 757:787 mass spectrometry (MS) analysis 757:787 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 7 43 from present 1451:1457 arg1 fraction 1471:1478 only one fraction 1462:1478 only one fraction 1462:1478 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 4 44 theme invasive 548:555 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 45 gly glycosylated 1805:1816 arg1 proteins 1818:1825 corresponding glycosylated proteins 1791:1825 corresponding glycosylated proteins 1791:1825 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 6 46 from ACA 1094:1096 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 46 from ACA 1094:1096 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 46 from ACA 1094:1096 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 5 47 theme protein 818:824 arg1 glycosylation 826:838 protein glycosylation 818:838 protein glycosylation of the four fractions 818:860 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 4 48 used used 388:391 arg2 We 385:386 We 385:386 We 385:386 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 49 theme glycomes 1649:1656 arg1 fraction 1670:1677 glycomes subcellular fraction 1649:1677 glycomes subcellular fraction 1649:1677 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 50 theme cells 618:622 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 51 from ConA 1075:1078 arg1 KK47 1065:1068 KK47 1065:1068 KK47 1065:1068 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 9 52 theme fraction 1670:1677 arg1 examination 1634:1644 detailed examination 1625:1644 detailed examination of glycomes subcellular fraction with high resolution and sensitivity 1625:1714 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 53 theme microsomal 625:634 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 54 from GNA 1081:1083 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 54 from GNA 1081:1083 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 54 from GNA 1081:1083 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 0 55 theme Distribution 39:50 arg1 Analysis 52:59 Differential Distribution Analysis 26:59 Differential Distribution Analysis 26:59 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 7 56 from N-glycans 1402:1410 arg1 YTS1 1286:1289 YTS1 1286:1289 YTS1 1286:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 56 from N-glycans 1402:1410 arg1 N-glycans 1318:1326 KK47 and 7 N-glycans 1307:1326 KK47 and 7 N-glycans in HCV29 were present in all four fractions 1307:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 56 from N-glycans 1402:1410 arg1 HCV29 1440:1444 HCV29 1440:1444 HCV29 1440:1444 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 56 from N-glycans 1402:1410 arg1 N-glycans 1380:1388 10 N-glycans 1377:1388 10 N-glycans in YTS1 1377:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 56 from N-glycans 1402:1410 arg1 KK47 1415:1418 KK47 1415:1418 KK47 1415:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 6 57 theme microarray 870:879 arg1 analysis 881:888 Lectin microarray analysis 863:888 Lectin microarray analysis 863:888 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 58 theme YTS1 1020:1023 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 9 59 theme high 1684:1687 arg1 resolution 1689:1698 high resolution 1684:1698 high resolution 1684:1698 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 60 theme normal 587:592 arg1 cells 618:622 normal bladder epithelia HCV29 cells 587:622 normal bladder epithelia HCV29 cells 587:622 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 61 dep nuclear 665:671 arg1 Nuc 674:676 Nuc 674:676 Nuc 674:676 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 62 theme centrifugation 406:419 arg1 techniques 421:430 differential centrifugation techniques 393:430 differential centrifugation techniques 393:430 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 63 theme epithelia 602:610 arg1 cells 618:622 normal bladder epithelia HCV29 cells 587:622 normal bladder epithelia HCV29 cells 587:622 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 2 64 theme Protein 235:241 arg1 glycosylation 243:255 Protein glycosylation 235:255 Protein glycosylation 235:255 Protein glycosylation provides a basis for diversity of protein functions. 27313494 4 65 dep cytosolic 684:692 arg1 Cyto 695:698 Cyto 695:698 Cyto 695:698 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 1 66 theme associated 169:178 arg1 processes 191:199 their associated biological processes 163:199 their associated biological processes 163:199 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 9 67 from organelles 1839:1848 arg1 elucidation 1740:1750 elucidation 1740:1750 elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles 1740:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 6 68 from terms 951:955 arg1 cell 1107:1110 HCV29 cell 1101:1110 HCV29 cell 1101:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 MPL 1000:1002 MPL 1000:1002 MPL 1000:1002 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 VVA 1086:1088 VVA 1086:1088 VVA 1086:1088 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 ACA 1094:1096 ACA 1094:1096 ACA 1094:1096 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 WGA 1005:1007 WGA 1005:1007 WGA 1005:1007 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 PWM 1013:1015 PWM 1013:1015 PWM 1013:1015 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 69 dep AAL 995:997 arg1 GNA 1081:1083 GNA 1081:1083 GNA 1081:1083 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 4 70 theme protein 460:466 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 71 from differences 911:921 arg1 terms 951:955 terms 951:955 terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell 951:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 8 72 theme latter 1496:1501 arg1 category 1503:1510 the latter category 1492:1510 the latter category 1492:1510 Glycans in the latter category are considered potential markers for the corresponding organelles. 27313494 7 73 from N-glycans 1144:1152 arg1 fractions 1162:1170 four fractions 1157:1170 four fractions of three cells detected by MS analysis 1157:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 8 74 theme potential 1527:1535 arg1 markers 1537:1543 potential markers 1527:1543 potential markers for the corresponding organelles 1527:1576 Glycans in the latter category are considered potential markers for the corresponding organelles. 27313494 7 75 gly fucosylated 1229:1239 arg1 structures 1241:1250 fucosylated structures 1229:1250 fucosylated structures 1229:1250 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 9 76 theme roles 1770:1774 arg1 elucidation 1740:1750 elucidation 1740:1750 elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles 1740:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 77 theme cytosolic 684:692 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 5 78 theme integrated 705:714 arg1 strategy 716:723 An integrated strategy 702:723 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis 702:787 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 7 79 attach present 1451:1457 arg2 N-glycans 1273:1281 10 N-glycans 1270:1281 10 N-glycans in YTS1 1270:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 79 attach present 1451:1457 arg2 N-glycans 1294:1302 5 N-glycans 1292:1302 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1 1292:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 79 attach present 1451:1457 arg2 N-glycans 1402:1410 16 N-glycans 1399:1410 16 N-glycans in KK47 1399:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 79 attach present 1451:1457 arg1 fraction 1471:1478 only one fraction 1462:1478 only one fraction 1462:1478 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 79 attach present 1451:1457 arg2 N-glycans 1427:1435 3 N-glycans 1425:1435 3 N-glycans 1425:1435 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 3 80 theme glycan 323:328 arg1 composition 330:340 glycan composition 323:340 glycan composition 323:340 Diversity of glycan composition in animal cells remains poorly understood. 27313494 4 81 theme cells 521:525 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 82 theme glycans 1779:1785 arg1 roles 1770:1774 the functional roles 1755:1774 the functional roles of glycans and corresponding glycosylated proteins in distinct organelles 1755:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 83 theme nuclear 665:671 arg1 fractions 468:476 four subcellular protein fractions 443:476 four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto) 443:699 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 8 84 theme corresponding 1553:1565 arg1 organelles 1567:1576 the corresponding organelles 1549:1576 the corresponding organelles 1549:1576 Glycans in the latter category are considered potential markers for the corresponding organelles. 27313494 5 85 theme fractions 852:860 arg1 glycosylation 826:838 protein glycosylation 818:838 protein glycosylation of the four fractions 818:860 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 4 86 theme low 528:530 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 7 87 from fraction 1471:1478 arg1 present 1451:1457 present 1451:1457 present 1451:1457 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 9 88 theme corresponding 1791:1803 arg1 proteins 1818:1825 corresponding glycosylated proteins 1791:1825 corresponding glycosylated proteins 1791:1825 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 6 89 theme glycan 960:965 arg1 binding 967:973 glycan binding 960:973 glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell 960:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 90 dep lectins 982:988 arg1 AAL 995:997 AAL 995:997 AAL 995:997 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 9 91 theme proteins 1818:1825 arg1 roles 1770:1774 the functional roles 1755:1774 the functional roles of glycans and corresponding glycosylated proteins in distinct organelles 1755:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 2 92 theme functions 299:307 arg1 diversity 278:286 diversity 278:286 diversity of protein functions 278:307 Protein glycosylation provides a basis for diversity of protein functions. 27313494 1 93 theme components 148:157 arg1 Compartmentalization 115:134 Compartmentalization 115:134 Compartmentalization of cellular components and their associated biological processes 115:199 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 9 94 with examination 1634:1644 arg1 sensitivity 1704:1714 sensitivity 1704:1714 sensitivity 1704:1714 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 9 94 with examination 1634:1644 arg1 resolution 1689:1698 high resolution 1684:1698 high resolution 1684:1698 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 95 theme bladder 508:514 arg1 cells 521:525 metastatic bladder YTS1 cells 497:525 metastatic bladder YTS1 cells 497:525 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 96 theme distinct 1830:1837 arg1 organelles 1839:1848 distinct organelles 1830:1848 distinct organelles 1830:1848 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 0 97 theme Subcellular 75:85 arg1 Fractions 87:95 Subcellular Fractions 75:95 Subcellular Fractions of Bladder Cells 75:112 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 4 98 theme KK47 572:575 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 99 from cell 1107:1110 arg1 terms 951:955 terms 951:955 terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell 951:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 1 100 theme biological 180:189 arg1 processes 191:199 their associated biological processes 163:199 their associated biological processes 163:199 Compartmentalization of cellular components and their associated biological processes is crucial for cellular function. 27313494 7 101 from N-glycans 1294:1302 arg1 YTS1 1286:1289 YTS1 1286:1289 YTS1 1286:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 101 from N-glycans 1294:1302 arg1 N-glycans 1318:1326 KK47 and 7 N-glycans 1307:1326 KK47 and 7 N-glycans in HCV29 were present in all four fractions 1307:1370 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 101 from N-glycans 1294:1302 arg1 HCV29 1440:1444 HCV29 1440:1444 HCV29 1440:1444 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 101 from N-glycans 1294:1302 arg1 N-glycans 1380:1388 10 N-glycans 1377:1388 10 N-glycans in YTS1 1377:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 101 from N-glycans 1294:1302 arg1 KK47 1415:1418 KK47 1415:1418 KK47 1415:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 0 102 from Identification 7:20 arg1 Fractions 87:95 Subcellular Fractions 75:95 Subcellular Fractions of Bladder Cells 75:112 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 6 103 from WGA 1005:1007 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 103 from WGA 1005:1007 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 103 from WGA 1005:1007 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 0 104 theme Cells 108:112 arg1 Fractions 87:95 Subcellular Fractions 75:95 Subcellular Fractions of Bladder Cells 75:112 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 4 105 theme nonmuscle 538:546 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 6 106 from PWM 1013:1015 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 106 from PWM 1013:1015 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 106 from PWM 1013:1015 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 5 107 theme mass 757:760 arg1 MS 776:777 MS 776:777 MS 776:777 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 5 107 theme mass 757:760 arg1 spectrometry 762:773 mass spectrometry 757:773 mass spectrometry (MS) analysis 757:787 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 4 108 dep microsomal 625:634 arg1 Mic 637:639 Mic 637:639 Mic 637:639 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 9 109 theme detailed 1625:1632 arg1 examination 1634:1644 detailed examination 1625:1644 detailed examination of glycomes subcellular fraction with high resolution and sensitivity 1625:1714 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 0 110 from Analysis 52:59 arg1 Fractions 87:95 Subcellular Fractions 75:95 Subcellular Fractions of Bladder Cells 75:112 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 6 111 from MPL 1000:1002 arg1 ConA 1075:1078 ConA 1075:1078 ConA 1075:1078 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 111 from MPL 1000:1002 arg1 STL 1031:1033 STL 1031:1033 STL 1031:1033 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 111 from MPL 1000:1002 arg1 cell 1025:1028 YTS1 cell 1020:1028 YTS1 cell 1020:1028 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 0 112 theme Differential 26:37 arg1 Analysis 52:59 Differential Distribution Analysis 26:59 Differential Distribution Analysis 26:59 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 7 113 located present 1451:1457 arg2 N-glycans 1273:1281 10 N-glycans 1270:1281 10 N-glycans in YTS1 1270:1289 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 113 located present 1451:1457 arg2 N-glycans 1294:1302 5 N-glycans 1292:1302 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1 1292:1396 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 113 located present 1451:1457 arg2 N-glycans 1402:1410 16 N-glycans 1399:1410 16 N-glycans in KK47 1399:1418 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 113 located present 1451:1457 arg1 fraction 1471:1478 only one fraction 1462:1478 only one fraction 1462:1478 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 113 located present 1451:1457 arg2 N-glycans 1427:1435 3 N-glycans 1425:1435 3 N-glycans 1425:1435 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 7 114 theme cells 1181:1185 arg1 fractions 1162:1170 four fractions 1157:1170 four fractions of three cells detected by MS analysis 1157:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 9 115 theme subcellular 1658:1668 arg1 fraction 1670:1677 glycomes subcellular fraction 1649:1677 glycomes subcellular fraction 1649:1677 The integrated strategy described here allows detailed examination of glycomes subcellular fraction with high resolution and sensitivity, and will be useful for elucidation of the functional roles of glycans and corresponding glycosylated proteins in distinct organelles. 27313494 4 116 theme bladder 594:600 arg1 cells 618:622 normal bladder epithelia HCV29 cells 587:622 normal bladder epithelia HCV29 cells 587:622 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 0 117 theme Glycans 64:70 arg1 Analysis 52:59 Differential Distribution Analysis 26:59 Differential Distribution Analysis 26:59 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 0 117 theme Glycans 64:70 arg1 Identification 7:20 Global Identification 0:20 Global Identification 0:20 Global Identification and Differential Distribution Analysis of Glycans in Subcellular Fractions of Bladder Cells. 27313494 6 118 theme Lectin 863:868 arg1 analysis 881:888 Lectin microarray analysis 863:888 Lectin microarray analysis 863:888 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 4 119 theme bladder 557:563 arg1 cells 577:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 low grade nonmuscle invasive bladder cancer KK47 cells 528:581 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 4 120 theme HCV29 612:616 arg1 cells 618:622 normal bladder epithelia HCV29 cells 587:622 normal bladder epithelia HCV29 cells 587:622 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 5 121 gly glycosylation 826:838 arg1 fractions 852:860 the four fractions 843:860 the four fractions 843:860 An integrated strategy combining lectin microarray and mass spectrometry (MS) analysis was then applied to evaluate protein glycosylation of the four fractions. 27313494 4 122 theme differential 393:404 arg1 techniques 421:430 differential centrifugation techniques 393:430 differential centrifugation techniques 393:430 We used differential centrifugation techniques to isolate four subcellular protein fractions from homogenate of metastatic bladder YTS1 cells, low grade nonmuscle invasive bladder cancer KK47 cells and normal bladder epithelia HCV29 cells: microsomal (Mic), mitochondrial (Mito), nuclear (Nuc), and cytosolic (Cyto). 27313494 7 123 theme N-glycans 1144:1152 arg1 total 1121:1125 a total 1119:1125 a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis 1119:1209 Among a total of 40, 32 and 15 N-glycans in four fractions of three cells detected by MS analysis, high-mannose and fucosylated structures were predominant, 10 N-glycans in YTS1, 5 N-glycans in KK47 and 7 N-glycans in HCV29 were present in all four fractions; and 10 N-glycans in YTS1, 16 N-glycans in KK47, and 3 N-glycans in HCV29 were present in only one fraction. 27313494 6 124 from binding 967:973 arg1 cell 1107:1110 HCV29 cell 1101:1110 HCV29 cell 1101:1110 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 27313494 6 125 from WGA 1058:1060 arg1 KK47 1065:1068 KK47 1065:1068 KK47 1065:1068 Lectin microarray analysis revealed significant differences among the four fractions in terms of glycan binding to the lectins LCA, AAL, MPL, WGA and PWM in YTS1 cell, STL, Jacalin, VVA, LCA and WGA in KK47, and ConA, GNA, VVA and ACA in HCV29 cell. 26921321 1 0 theme spore 320:324 arg1 adhesion 326:333 spore adhesion 320:333 spore adhesion 320:333 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 6 1 theme collagen-like 1019:1031 arg1 regions 1033:1039 the collagen-like regions 1015:1039 the collagen-like regions of both B. anthracis and B. cereus 1015:1074 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 1 2 theme paracrystalline 230:244 arg1 layer 269:273 a paracrystalline flexible yet resistant layer 228:273 a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence 228:347 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 5 3 theme BclA 936:939 arg1 glycosylation 941:953 BclA glycosylation 936:953 BclA glycosylation 936:953 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 8 4 theme carbohydrate 1439:1450 arg1 polymers 1452:1459 large carbohydrate polymers 1433:1459 large carbohydrate polymers covering the surface of Bacillus spores 1433:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 1 5 theme flexible 246:253 arg1 layer 269:273 a paracrystalline flexible yet resistant layer 228:273 a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence 228:347 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 8 6 theme large 1433:1437 arg1 polymers 1452:1459 large carbohydrate polymers 1433:1459 large carbohydrate polymers covering the surface of Bacillus spores 1433:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 1 7 dep Bacillus 128:135 arg1 cereus 137:142 cereus 137:142 cereus 137:142 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 4 8 theme collagen-like 637:649 arg1 region 651:656 The collagen-like region 633:656 The collagen-like region of B. anthracis 633:672 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 3 9 gly glycoprotein 605:616 arg1 glycoprotein 605:616 glycoprotein trimerization 605:630 glycoprotein trimerization 605:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 5 10 theme domain 914:919 arg1 specificity 921:931 the species and domain specificity 898:931 specificity 921:931 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 8 11 with interactions 1566:1577 arg1 surfaces 1603:1610 biotic and abiotic surfaces 1584:1610 biotic and abiotic surfaces 1584:1610 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 2 12 attach attached 434:441 arg2 constituent 360:370 The major constituent 350:370 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, 350:429 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 12 attach attached 434:441 arg1 layer 456:460 the basal layer 446:460 the basal layer 446:460 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 8 13 theme abiotic 1595:1601 arg1 surfaces 1603:1610 biotic and abiotic surfaces 1584:1610 biotic and abiotic surfaces 1584:1610 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 1 14 theme resistant 259:267 arg1 layer 269:273 a paracrystalline flexible yet resistant layer 228:273 a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence 228:347 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 0 15 from cereus 49:54 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium 0:88 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 4 16 theme targeted 823:830 arg1 interventions 844:856 thus targeted therapeutic interventions 818:856 thus targeted therapeutic interventions 818:856 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 6 17 theme anthracis 1052:1060 arg1 regions 1033:1039 the collagen-like regions 1015:1039 the collagen-like regions of both B. anthracis and B. cereus 1015:1074 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 18 theme deoxyhexose 1152:1162 arg1 residues 1164:1171 the species-specific deoxyhexose residues 1131:1171 the species-specific deoxyhexose residues anthrose and the newly observed cereose 1131:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 18 theme deoxyhexose 1152:1162 arg1 cereose 1205:1211 the newly observed cereose 1186:1211 the newly observed cereose 1186:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 18 theme deoxyhexose 1152:1162 arg1 anthrose 1173:1180 anthrose 1173:1180 anthrose 1173:1180 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 2 19 theme hairlike 379:386 arg1 BclA 425:428 the trimerized glycoprotein BclA 397:428 the trimerized glycoprotein BclA 397:428 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 19 theme hairlike 379:386 arg1 surface 388:394 its hairlike surface 375:394 its hairlike surface 375:394 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 5 20 theme B. 966:967 arg1 group 976:980 the B. cereus group 962:980 the B. cereus group 962:980 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 4 21 theme unusual 712:718 arg1 O-glycans 720:728 unusual O-glycans 712:728 unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions 712:856 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 4 22 theme therapeutic 832:842 arg1 interventions 844:856 thus targeted therapeutic interventions 818:856 thus targeted therapeutic interventions 818:856 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 7 23 theme O-linked 1356:1363 arg1 glycans 1365:1371 polysaccharide-like O-linked glycans 1336:1371 polysaccharide-like O-linked glycans whose structures are also species-specific 1336:1414 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 5 24 theme present 866:872 arg1 study 874:878 the present study 862:878 the present study 862:878 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 0 25 theme Glycoprotein 22:33 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium 0:88 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 3 26 theme C-terminal 572:581 arg1 head 566:569 a globular head 555:569 a globular head (C-terminal domain) that promotes glycoprotein trimerization 555:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 3 26 theme C-terminal 572:581 arg1 domain 583:588 C-terminal domain 572:588 C-terminal domain 572:588 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 6 27 dep residues 1164:1171 arg1 residues 1164:1171 the species-specific deoxyhexose residues 1131:1171 the species-specific deoxyhexose residues anthrose and the newly observed cereose 1131:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 27 dep residues 1164:1171 arg1 cereose 1205:1211 the newly observed cereose 1186:1211 the newly observed cereose 1186:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 27 dep residues 1164:1171 arg1 anthrose 1173:1180 anthrose 1173:1180 anthrose 1173:1180 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 4 28 theme spores 807:812 arg1 diagnostics 779:789 species-specific diagnostics 762:789 species-specific diagnostics of B. anthracis spores 762:812 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 4 28 theme spores 807:812 arg1 interventions 844:856 thus targeted therapeutic interventions 818:856 thus targeted therapeutic interventions 818:856 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 1 29 dep group 144:148 arg1 anthracis 171:179 Bacillus anthracis 162:179 Bacillus anthracis 162:179 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 1 29 dep group 144:148 arg1 thuringiensis 195:207 Bacillus thuringiensis 186:207 Bacillus thuringiensis 186:207 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 1 29 dep group 144:148 arg1 cereus 154:159 B. cereus 151:159 B. cereus 151:159 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 2 30 theme trimerized 401:410 arg1 BclA 425:428 the trimerized glycoprotein BclA 397:428 the trimerized glycoprotein BclA 397:428 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 30 theme trimerized 401:410 arg1 surface 388:394 its hairlike surface 375:394 its hairlike surface 375:394 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 31 theme glycoprotein 412:423 arg1 BclA 425:428 the trimerized glycoprotein BclA 397:428 the trimerized glycoprotein BclA 397:428 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 31 theme glycoprotein 412:423 arg1 surface 388:394 its hairlike surface 375:394 its hairlike surface 375:394 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 8 32 theme polymers 1452:1459 arg1 presence 1421:1428 The presence 1417:1428 The presence of large carbohydrate polymers covering the surface of Bacillus spores 1417:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 3 33 theme collagen-like 528:540 arg1 neck 542:545 a repetitive collagen-like neck 515:545 a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization 515:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 2 34 theme major 354:358 arg1 constituent 360:370 The major constituent 350:370 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, 350:429 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 2 35 theme basal 450:454 arg1 layer 456:460 the basal layer 446:460 the basal layer 446:460 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 7 36 theme globular 1274:1281 arg1 BclA 1294:1297 BclA 1294:1297 BclA 1294:1297 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 7 36 theme globular 1274:1281 arg1 domains 1283:1289 the C-terminal globular domains 1259:1289 the C-terminal globular domains of BclA from both species 1259:1315 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 0 37 gly Glycosylation 0:12 arg1 Glycoprotein 22:33 BclA Glycoprotein 17:33 BclA Glycoprotein 17:33 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 4 38 theme anthracis 664:672 arg1 region 651:656 The collagen-like region 633:656 The collagen-like region of B. anthracis 633:672 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 8 39 theme diagnostic 1641:1650 arg1 targets 1652:1658 potential new diagnostic targets 1627:1658 potential new diagnostic targets 1627:1658 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 0 40 from anthracis 69:77 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium 0:88 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 0 41 dep cereus 49:54 arg1 Exosporium 79:88 Exosporium 79:88 Exosporium 79:88 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 7 42 from domains 1283:1289 arg1 species 1309:1315 both species 1304:1315 both species 1304:1315 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 2 43 theme surface 388:394 arg1 constituent 360:370 The major constituent 350:370 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, 350:429 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 7 44 link O-linked 1356:1363 arg1 glycans 1365:1371 polysaccharide-like O-linked glycans 1336:1371 polysaccharide-like O-linked glycans whose structures are also species-specific 1336:1414 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 8 45 theme biotic 1584:1589 arg1 surfaces 1603:1610 biotic and abiotic surfaces 1584:1610 biotic and abiotic surfaces 1584:1610 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 3 46 theme glycoprotein 605:616 arg1 trimerization 618:630 glycoprotein trimerization 605:630 glycoprotein trimerization 605:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 6 47 theme short 1105:1109 arg1 O-glycans 1111:1119 short O-glycans 1105:1119 short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively 1105:1225 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 7 48 from species 1309:1315 arg1 BclA 1294:1297 BclA 1294:1297 BclA 1294:1297 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 7 48 from species 1309:1315 arg1 domains 1283:1289 the C-terminal globular domains 1259:1289 the C-terminal globular domains of BclA from both species 1259:1315 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 1 49 theme major 306:310 arg1 role 312:315 a major role 304:315 a major role 304:315 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 8 50 theme spores 1494:1499 arg1 surface 1474:1480 the surface 1470:1480 the surface of Bacillus spores 1470:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 3 51 theme globular 557:564 arg1 head 566:569 a globular head 555:569 a globular head (C-terminal domain) that promotes glycoprotein trimerization 555:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 3 51 theme globular 557:564 arg1 domain 583:588 C-terminal domain 572:588 C-terminal domain 572:588 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 7 52 theme C-terminal 1263:1272 arg1 BclA 1294:1297 BclA 1294:1297 BclA 1294:1297 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 7 52 theme C-terminal 1263:1272 arg1 domains 1283:1289 the C-terminal globular domains 1259:1289 the C-terminal globular domains of BclA from both species 1259:1315 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 5 53 dep B. 966:967 arg1 cereus 969:974 cereus 969:974 cereus 969:974 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 4 54 used used 742:745 arg2 O-glycans 720:728 unusual O-glycans 712:728 unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions 712:856 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 7 55 theme polysaccharide-like 1336:1354 arg1 glycans 1365:1371 polysaccharide-like O-linked glycans 1336:1371 polysaccharide-like O-linked glycans whose structures are also species-specific 1336:1414 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 6 56 theme observed 1196:1203 arg1 residues 1164:1171 the species-specific deoxyhexose residues 1131:1171 the species-specific deoxyhexose residues anthrose and the newly observed cereose 1131:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 56 theme observed 1196:1203 arg1 cereose 1205:1211 the newly observed cereose 1186:1211 the newly observed cereose 1186:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 4 57 theme species-specific 762:777 arg1 diagnostics 779:789 species-specific diagnostics 762:789 species-specific diagnostics of B. anthracis spores 762:812 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 4 58 theme B. 794:795 arg1 spores 807:812 B. anthracis spores 794:812 B. anthracis spores 794:812 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 1 59 theme Bacillus 128:135 arg1 group 144:148 the Bacillus cereus group 124:148 the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) 124:208 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 3 60 theme repetitive 517:526 arg1 neck 542:545 a repetitive collagen-like neck 515:545 a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization 515:630 It is then followed by a repetitive collagen-like neck bearing a globular head (C-terminal domain) that promotes glycoprotein trimerization. 26921321 6 61 theme species-specific 1135:1150 arg1 residues 1164:1171 the species-specific deoxyhexose residues 1131:1171 the species-specific deoxyhexose residues anthrose and the newly observed cereose 1131:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 61 theme species-specific 1135:1150 arg1 cereose 1205:1211 the newly observed cereose 1186:1211 the newly observed cereose 1186:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 61 theme species-specific 1135:1150 arg1 anthrose 1173:1180 anthrose 1173:1180 anthrose 1173:1180 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 5 62 theme glycosylation 941:953 arg1 species 902:908 the species and domain specificity 898:931 species 902:908 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 5 62 theme glycosylation 941:953 arg1 specificity 921:931 the species and domain specificity 898:931 specificity 921:931 In the present study, we have explored the species and domain specificity of BclA glycosylation within the B. cereus group. 26921321 8 63 theme Bacillus 1485:1492 arg1 spores 1494:1499 Bacillus spores 1485:1499 Bacillus spores 1485:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 2 64 theme N-terminal 473:482 arg1 domain 484:489 an N-terminal domain 470:489 an N-terminal domain 470:489 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 4 65 dep B. 794:795 arg1 anthracis 797:805 anthracis 797:805 anthracis 797:805 The collagen-like region of B. anthracis is known to be densely substituted by unusual O-glycans that may be used for developing species-specific diagnostics of B. anthracis spores and thus targeted therapeutic interventions. 26921321 2 66 gly glycoprotein 412:423 arg1 glycoprotein 412:423 the trimerized glycoprotein BclA 397:428 the trimerized glycoprotein BclA 397:428 The major constituent of its hairlike surface, the trimerized glycoprotein BclA, is attached to the basal layer through an N-terminal domain. 26921321 7 67 theme BclA 1294:1297 arg1 BclA 1294:1297 BclA 1294:1297 BclA 1294:1297 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 7 67 theme BclA 1294:1297 arg1 domains 1283:1289 the C-terminal globular domains 1259:1289 the C-terminal globular domains of BclA from both species 1259:1315 Second we have discovered that the C-terminal globular domains of BclA from both species are substituted by polysaccharide-like O-linked glycans whose structures are also species-specific. 26921321 1 68 theme group 144:148 arg1 spores 114:119 The spores 110:119 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) 110:208 The spores of the Bacillus cereus group (B. cereus, Bacillus anthracis, and Bacillus thuringiensis) are surrounded by a paracrystalline flexible yet resistant layer called exosporium that plays a major role in spore adhesion and virulence. 26921321 8 69 contain have 1505:1508 arg1 presence 1421:1428 The presence 1417:1428 The presence of large carbohydrate polymers covering the surface of Bacillus spores 1417:1499 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 8 69 contain have 1505:1508 arg2 impact 1521:1526 a profound impact 1510:1526 a profound impact 1510:1526 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 6 70 contain bear 1126:1129 arg2 anthrose 1173:1180 anthrose 1173:1180 anthrose 1173:1180 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 70 contain bear 1126:1129 arg1 O-glycans 1111:1119 short O-glycans 1105:1119 short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively 1105:1225 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 70 contain bear 1126:1129 arg2 residues 1164:1171 the species-specific deoxyhexose residues 1131:1171 the species-specific deoxyhexose residues anthrose and the newly observed cereose 1131:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 6 70 contain bear 1126:1129 arg2 cereose 1205:1211 the newly observed cereose 1186:1211 the newly observed cereose 1186:1211 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 8 71 theme profound 1512:1519 arg1 impact 1521:1526 a profound impact 1510:1526 a profound impact 1510:1526 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 26921321 0 72 theme BclA 17:20 arg1 Glycoprotein 22:33 BclA Glycoprotein 17:33 BclA Glycoprotein 17:33 Glycosylation of BclA Glycoprotein from Bacillus cereus and Bacillus anthracis Exosporium Is Domain-specific. 26921321 6 73 theme cereus 1069:1074 arg1 regions 1033:1039 the collagen-like regions 1015:1039 the collagen-like regions of both B. anthracis and B. cereus 1015:1074 First, we have established that the collagen-like regions of both B. anthracis and B. cereus are similarly substituted by short O-glycans that bear the species-specific deoxyhexose residues anthrose and the newly observed cereose, respectively. 26921321 8 74 theme potential 1627:1635 arg1 targets 1652:1658 potential new diagnostic targets 1627:1658 potential new diagnostic targets 1627:1658 The presence of large carbohydrate polymers covering the surface of Bacillus spores may have a profound impact on the way that spores regulate their interactions with biotic and abiotic surfaces and represents potential new diagnostic targets. 27639389 4 0 theme spectrometry 1032:1043 arg1 approach 1077:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 1 1 theme antigens 213:220 arg1 expression 166:175 the expression 162:175 the expression of A, B or H antigens and the Lewis antigens 162:220 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 4 2 theme BEC 840:842 arg1 proteins 853:860 BEC membrane proteins 840:860 BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 840:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 0 3 with interaction 58:68 arg1 cells 93:97 buccal epithelial cells 75:97 buccal epithelial cells 75:97 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 7 4 theme similar 1478:1484 arg1 types 1493:1497 similar glycan types 1478:1497 similar glycan types 1478:1497 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 4 5 theme antigens 800:807 arg1 expression 758:767 the expression 754:767 the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 754:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 6 theme chromatography 988:1001 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 6 theme chromatography 988:1001 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 7 theme N-and 812:816 arg1 glycans 827:833 N-and O-linked glycans 812:833 N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 812:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 7 8 theme N-glycan 1421:1428 arg1 profiles 1430:1437 the N-glycan profiles 1417:1437 the N-glycan profiles of BEC from all blood groups 1417:1466 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 9 9 theme Candida 1766:1772 arg1 albicans 1774:1781 the oral pathogen Candida albicans 1748:1781 the oral pathogen Candida albicans 1748:1781 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 9 10 theme adhesion 1732:1739 arg1 assays 1741:1746 adhesion assays 1732:1746 adhesion assays 1732:1746 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 4 11 link O-linked 818:825 arg1 glycans 827:833 N-and O-linked glycans 812:833 N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 812:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 12 theme different 902:910 arg1 type 924:927 different blood group type 902:927 different blood group type 902:927 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 8 13 from individuals 1618:1628 arg1 analysis 1587:1594 multivariate analysis 1574:1594 multivariate analysis of the O-glycans from individuals 1574:1628 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 8 13 from individuals 1618:1628 arg1 O-glycans 1603:1611 the O-glycans 1599:1611 the O-glycans from individuals 1599:1628 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 3 14 theme group 586:590 arg1 antigens 592:599 blood group antigens 580:599 blood group antigens 580:599 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 9 15 theme group 1834:1838 arg1 BECs 1847:1850 blood group O type BECs 1828:1850 blood group O type BECs relative to other blood groups 1828:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 9 16 theme pathogen 1757:1764 arg1 albicans 1774:1781 the oral pathogen Candida albicans 1748:1781 the oral pathogen Candida albicans 1748:1781 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 2 17 from presentation 474:485 arg1 surface 543:549 the cell surface 534:549 the cell surface 534:549 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 6 18 theme blood 1318:1322 arg1 determinants 1330:1341 any A/B blood group determinants 1310:1341 any A/B blood group determinants 1310:1341 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 1 19 theme group 112:116 arg1 polymorphisms 118:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 4 20 theme various 865:871 arg1 individuals 873:883 various individuals 865:883 various individuals 865:883 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 21 from proteins 853:860 arg1 glycans 827:833 N-and O-linked glycans 812:833 N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 812:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 9 22 theme type 1842:1845 arg1 BECs 1847:1850 blood group O type BECs 1828:1850 blood group O type BECs relative to other blood groups 1828:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 9 23 theme higher 1806:1811 arg1 interaction 1813:1823 a significantly higher interaction 1790:1823 a significantly higher interaction to blood group O type BECs relative to other blood groups 1790:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 2 24 from inheritance 288:298 arg1 genes 320:324 genes 320:324 genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 320:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 2 25 theme antigens 522:529 arg1 presentation 474:485 the presentation 470:485 the presentation of different glycosylated terminal antigens on the cell surface 470:549 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 0 26 theme epithelial 82:91 arg1 cells 93:97 buccal epithelial cells 75:97 buccal epithelial cells 75:97 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 2 27 theme glycosylated 500:511 arg1 antigens 522:529 different glycosylated terminal antigens 490:529 different glycosylated terminal antigens 490:529 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 6 28 theme several 1358:1364 arg1 H-antigens 1375:1384 several terminal H-antigens 1358:1384 several terminal H-antigens 1358:1384 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 5 29 theme BEC 1098:1100 arg1 proteins 1111:1118 these BEC membrane proteins 1092:1118 these BEC membrane proteins 1092:1118 From these BEC membrane proteins a total of 77 N-glycan and 96 O-glycan structures were structurally characterized from 19 individuals and relatively quantitated. 27639389 9 30 theme other 1864:1868 arg1 groups 1876:1881 other blood groups 1864:1881 other blood groups 1864:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 2 31 gly glycosylated 500:511 arg1 antigens 522:529 different glycosylated terminal antigens 490:529 different glycosylated terminal antigens 490:529 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 4 32 theme electrospray 1003:1014 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 32 theme electrospray 1003:1014 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 0 33 theme Blood 0:4 arg1 expression 20:29 Blood group antigen expression 0:29 Blood group antigen expression 0:29 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 5 34 theme structures 1159:1168 arg1 total 1122:1126 a total 1120:1126 a total of 77 N-glycan and 96 O-glycan structures 1120:1168 From these BEC membrane proteins a total of 77 N-glycan and 96 O-glycan structures were structurally characterized from 19 individuals and relatively quantitated. 27639389 4 35 theme mass 1027:1030 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 35 theme mass 1027:1030 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 9 36 theme blood 1828:1832 arg1 BECs 1847:1850 blood group O type BECs 1828:1850 blood group O type BECs relative to other blood groups 1828:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 9 37 theme relative 1852:1859 arg1 BECs 1847:1850 blood group O type BECs 1828:1850 blood group O type BECs relative to other blood groups 1828:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 2 38 dep genes 320:324 arg1 regulating 339:348 regulating 339:348 regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 339:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 2 38 dep genes 320:324 arg1 encoding 326:333 encoding 326:333 encoding 326:333 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 2 39 theme bodily 394:399 arg1 secretions 401:410 bodily secretions 394:410 bodily secretions 394:410 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 0 40 theme antigen 12:18 arg1 expression 20:29 Blood group antigen expression 0:29 Blood group antigen expression 0:29 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 1 41 theme Human 100:104 arg1 polymorphisms 118:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 4 42 theme graphitic 964:972 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 42 theme graphitic 964:972 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 8 43 theme multivariate 1574:1585 arg1 analysis 1587:1594 multivariate analysis 1574:1594 multivariate analysis of the O-glycans from individuals 1574:1628 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 3 44 theme epithelial 645:654 arg1 BEC 663:665 BEC 663:665 BEC 663:665 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 3 44 theme epithelial 645:654 arg1 cells 656:660 buccal epithelial cells 638:660 buccal epithelial cells (BEC) that line the oral cavity 638:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 2 45 theme antigens 382:389 arg1 expression 354:363 the expression 350:363 the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 350:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 5 46 theme 77 N-glycan 1131:1141 arg1 total 1122:1126 a total 1120:1126 a total of 77 N-glycan and 96 O-glycan structures 1120:1168 From these BEC membrane proteins a total of 77 N-glycan and 96 O-glycan structures were structurally characterized from 19 individuals and relatively quantitated. 27639389 9 47 theme blood 1870:1874 arg1 groups 1876:1881 other blood groups 1864:1881 other blood groups 1864:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 4 48 theme liquid 981:986 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 48 theme liquid 981:986 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 49 theme glycomics 1067:1075 arg1 approach 1077:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 8 50 theme O-glycans 1603:1611 arg1 analysis 1587:1594 multivariate analysis 1574:1594 multivariate analysis of the O-glycans from individuals 1574:1628 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 4 51 theme group 784:788 arg1 antigens 800:807 histo-blood group (A, B, O) antigens 772:807 antigens 800:807 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 2 52 theme polymorphisms 303:315 arg1 inheritance 288:298 inheritance 288:298 inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 288:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 8 53 theme blood 1691:1695 arg1 type 1703:1706 their blood group type and secretor status 1685:1726 type 1703:1706 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 1 54 theme H 188:188 arg1 antigens 190:197 A, B or H antigens 180:197 A, B or H antigens 180:197 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 9 55 theme O 1840:1840 arg1 BECs 1847:1850 blood group O type BECs 1828:1850 blood group O type BECs relative to other blood groups 1828:1881 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 4 56 theme A 791:791 arg1 antigens 800:807 histo-blood group (A, B, O) antigens 772:807 antigens 800:807 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 8 57 theme segregation 1640:1650 arg1 patterns 1652:1659 segregation patterns 1640:1659 segregation patterns clearly associated with their blood group type and secretor status 1640:1726 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 3 58 theme oral 682:685 arg1 cavity 687:692 the oral cavity 678:692 the oral cavity 678:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 8 59 from analysis 1587:1594 arg1 individuals 1618:1628 individuals 1618:1628 individuals 1618:1628 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 8 60 theme secretor 1712:1719 arg1 status 1721:1726 their blood group type and secretor status 1685:1726 status 1721:1726 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 1 61 theme Lewis 207:211 arg1 antigens 213:220 the Lewis antigens 203:220 the Lewis antigens 203:220 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 7 62 theme glycan 1486:1491 arg1 types 1493:1497 similar glycan types 1478:1497 similar glycan types 1478:1497 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 4 63 theme O-linked 818:825 arg1 glycans 827:833 N-and O-linked glycans 812:833 N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 812:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 2 64 from polymorphisms 303:315 arg1 genes 320:324 genes 320:324 genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 320:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 2 65 from genes 320:324 arg1 inheritance 288:298 inheritance 288:298 inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces 288:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 3 66 gly glycosylation 621:633 arg1 BEC 663:665 BEC 663:665 BEC 663:665 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 3 66 gly glycosylation 621:633 arg1 cells 656:660 buccal epithelial cells 638:660 buccal epithelial cells (BEC) that line the oral cavity 638:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 6 67 theme secretor 1273:1280 arg1 individuals 1282:1292 the secretor individuals 1269:1292 the secretor individuals 1269:1292 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 2 68 theme microbial 242:250 arg1 infections 252:261 microbial infections 242:261 microbial infections 242:261 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 7 69 theme blood 1455:1459 arg1 groups 1461:1466 all blood groups 1451:1466 all blood groups 1451:1466 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 7 70 theme BEC 1442:1444 arg1 profiles 1430:1437 the N-glycan profiles 1417:1437 the N-glycan profiles of BEC from all blood groups 1417:1466 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 7 71 from groups 1461:1466 arg1 profiles 1430:1437 the N-glycan profiles 1417:1437 the N-glycan profiles of BEC from all blood groups 1417:1466 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 1 72 theme A 180:180 arg1 antigens 190:197 A, B or H antigens 180:197 A, B or H antigens 180:197 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 4 73 theme individuals 873:883 arg1 proteins 853:860 BEC membrane proteins 840:860 BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 840:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 3 74 theme blood 580:584 arg1 antigens 592:599 blood group antigens 580:599 blood group antigens 580:599 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 1 75 theme B 183:183 arg1 antigens 190:197 A, B or H antigens 180:197 A, B or H antigens 180:197 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 9 76 theme oral 1752:1755 arg1 albicans 1774:1781 the oral pathogen Candida albicans 1748:1781 the oral pathogen Candida albicans 1748:1781 In adhesion assays the oral pathogen Candida albicans showed a significantly higher interaction to blood group O type BECs relative to other blood groups. 27639389 4 77 theme group 918:922 arg1 type 924:927 different blood group type 902:927 different blood group type 902:927 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 6 78 theme group 1324:1328 arg1 determinants 1330:1341 any A/B blood group determinants 1310:1341 any A/B blood group determinants 1310:1341 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 1 79 theme blood 106:110 arg1 polymorphisms 118:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms 100:130 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 4 80 theme membrane 844:851 arg1 proteins 853:860 BEC membrane proteins 840:860 BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 840:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 6 81 theme A/B 1314:1316 arg1 determinants 1330:1341 any A/B blood group determinants 1310:1341 any A/B blood group determinants 1310:1341 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 3 82 theme antigens 592:599 arg1 role 572:575 the role 568:575 the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity 568:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 6 83 contain contained 1348:1356 arg1 N-glycans 1254:1262 The N-glycans 1250:1262 The N-glycans from the secretor individuals 1250:1292 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 6 83 contain contained 1348:1356 arg2 H-antigens 1375:1384 several terminal H-antigens 1358:1384 several terminal H-antigens 1358:1384 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 4 84 theme secretor 933:940 arg1 status 942:947 secretor status 933:947 secretor status 933:947 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 2 85 theme terminal 513:520 arg1 antigens 522:529 different glycosylated terminal antigens 490:529 different glycosylated terminal antigens 490:529 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 0 86 theme buccal 75:80 arg1 cells 93:97 buccal epithelial cells 75:97 buccal epithelial cells 75:97 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 2 87 theme cell 538:541 arg1 surface 543:549 the cell surface 534:549 the cell surface 534:549 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 4 88 from expression 758:767 arg1 glycans 827:833 N-and O-linked glycans 812:833 N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 812:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 89 theme blood 912:916 arg1 type 924:927 different blood group type 902:927 different blood group type 902:927 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 6 90 theme terminal 1366:1373 arg1 H-antigens 1375:1384 several terminal H-antigens 1358:1384 several terminal H-antigens 1358:1384 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 2 91 theme different 490:498 arg1 antigens 522:529 different glycosylated terminal antigens 490:529 different glycosylated terminal antigens 490:529 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 6 92 from individuals 1282:1292 arg1 N-glycans 1254:1262 The N-glycans 1250:1262 The N-glycans from the secretor individuals 1250:1292 The N-glycans from the secretor individuals did not express any A/B blood group determinants, but contained several terminal H-antigens. 27639389 2 93 theme ABH 368:370 arg1 antigens 382:389 ABH and Lewis antigens 368:389 antigens 382:389 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 0 94 theme group 6:10 arg1 expression 20:29 Blood group antigen expression 0:29 Blood group antigen expression 0:29 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 5 95 theme membrane 1102:1109 arg1 proteins 1111:1118 these BEC membrane proteins 1092:1118 these BEC membrane proteins 1092:1118 From these BEC membrane proteins a total of 77 N-glycan and 96 O-glycan structures were structurally characterized from 19 individuals and relatively quantitated. 27639389 2 96 theme epithelial 416:425 arg1 surfaces 434:441 epithelial tissue surfaces 416:441 epithelial tissue surfaces 416:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 4 97 dep antigens 800:807 arg1 B 794:794 histo-blood group (A, B, O) antigens 772:807 B 794:794 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 97 dep antigens 800:807 arg1 O 797:797 histo-blood group (A, B, O) antigens 772:807 O 797:797 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 98 theme ionization 1016:1025 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 98 theme ionization 1016:1025 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 3 99 theme buccal 638:643 arg1 BEC 663:665 BEC 663:665 BEC 663:665 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 3 99 theme buccal 638:643 arg1 cells 656:660 buccal epithelial cells 638:660 buccal epithelial cells (BEC) that line the oral cavity 638:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 0 100 dep C. 46:47 arg1 albicans 49:56 albicans 49:56 albicans 49:56 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 4 101 theme porous 957:962 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 101 theme porous 957:962 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 2 102 theme Lewis 376:380 arg1 antigens 382:389 ABH and Lewis antigens 368:389 antigens 382:389 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 0 103 theme C. 46:47 arg1 interaction 58:68 C. albicans interaction 46:68 C. albicans interaction with buccal epithelial cells 46:97 Blood group antigen expression is involved in C. albicans interaction with buccal epithelial cells. 27639389 2 104 from expression 354:363 arg1 secretions 401:410 bodily secretions 394:410 bodily secretions 394:410 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 2 104 from expression 354:363 arg1 surfaces 434:441 epithelial tissue surfaces 416:441 epithelial tissue surfaces 416:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 27639389 4 105 theme carbon 974:979 arg1 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 PGC-LC-ESI-MS 1046:1058 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 4 105 theme carbon 974:979 arg1 spectrometry 1032:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry 955:1043 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 8 106 theme group 1697:1701 arg1 type 1703:1706 their blood group type and secretor status 1685:1726 type 1703:1706 However, multivariate analysis of the O-glycans from individuals displayed segregation patterns clearly associated with their blood group type and secretor status. 27639389 5 107 theme O-glycan 1150:1157 arg1 structures 1159:1168 96 O-glycan structures 1147:1168 96 O-glycan structures 1147:1168 From these BEC membrane proteins a total of 77 N-glycan and 96 O-glycan structures were structurally characterized from 19 individuals and relatively quantitated. 27639389 7 108 theme relative 1523:1530 arg1 intensities 1532:1542 their relative intensities 1517:1542 their relative intensities between individuals 1517:1562 Apart from the non-secretors, the N-glycan profiles of BEC from all blood groups displayed similar glycan types, while varying in their relative intensities between individuals. 27639389 4 109 theme based 1061:1065 arg1 approach 1077:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach 955:1084 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 3 110 theme cells 656:660 arg1 glycosylation 621:633 the glycosylation 617:633 the glycosylation of buccal epithelial cells (BEC) that line the oral cavity 617:692 We investigated the role of blood group antigens in diversifying the glycosylation of buccal epithelial cells (BEC) that line the oral cavity. 27639389 4 111 theme histo-blood 772:782 arg1 antigens 800:807 histo-blood group (A, B, O) antigens 772:807 antigens 800:807 Specifically, we characterized and statistically evaluated the expression of histo-blood group (A, B, O) antigens on N-and O-linked glycans from BEC membrane proteins of various individuals that represented different blood group type and secretor status using a porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry (PGC-LC-ESI-MS) based glycomics approach. 27639389 1 112 theme antigens 190:197 arg1 expression 166:175 the expression 162:175 the expression of A, B or H antigens and the Lewis antigens 162:220 Human blood group polymorphisms are known to be determined by the expression of A, B or H antigens and the Lewis antigens. 27639389 2 113 theme tissue 427:432 arg1 surfaces 434:441 epithelial tissue surfaces 416:441 epithelial tissue surfaces 416:441 Protection against microbial infections has been associated with inheritance of polymorphisms in genes encoding and regulating the expression of ABH and Lewis antigens in bodily secretions and epithelial tissue surfaces, subsequently resulting in the presentation of different glycosylated terminal antigens on the cell surface. 24644239 6 0 theme glycoproteins 903:915 arg1 assays 887:892 Chemotaxis assays 876:892 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins 876:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 1 1 theme glycosylated 151:162 arg1 CCL1 131:134 CCL1 131:134 CCL1 131:134 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 1 theme glycosylated 151:162 arg1 protein 174:180 a naturally glycosylated chemokine protein 139:180 a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes 139:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 1 theme glycosylated 151:162 arg1 chemoattractant 234:248 a chemoattractant 232:248 a chemoattractant for monocytes 232:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 8 2 theme homogeneous 1231:1241 arg1 molecules 1266:1274 homogeneous N-glycosylated protein molecules 1231:1274 homogeneous N-glycosylated protein molecules of defined covalent structure 1231:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 2 3 contain having 324:329 arg2 site 351:354 one N-glycosylation site 331:354 one N-glycosylation site 331:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 3 contain having 324:329 arg1 protein 316:322 a 73 amino acid protein 300:322 a 73 amino acid protein having one N-glycosylation site 300:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 3 contain having 324:329 arg1 CCL1 277:280 CCL1 277:280 CCL1 277:280 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 4 4 gly non-glycosylated 631:646 arg1 CCL1 663:666 (Ser-)CCL1 657:666 (Ser-)CCL1 657:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 4 4 gly non-glycosylated 631:646 arg1 forms 648:652 both N-glycosylated and non-glycosylated forms 607:652 both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 607:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 1 5 theme chemokine 164:172 arg1 CCL1 131:134 CCL1 131:134 CCL1 131:134 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 5 theme chemokine 164:172 arg1 protein 174:180 a naturally glycosylated chemokine protein 139:180 a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes 139:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 5 theme chemokine 164:172 arg1 chemoattractant 234:248 a chemoattractant 232:248 a chemoattractant for monocytes 232:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 6 6 theme proteins 956:963 arg1 assays 887:892 Chemotaxis assays 876:892 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins 876:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 7 7 theme protein 1054:1060 arg1 molecules 1062:1070 the (glyco)protein molecules 1043:1070 the (glyco)protein molecules 1043:1070 The results were correlated with the chemical structures of the (glyco)protein molecules. 24644239 2 8 gly non-glycosylated 382:397 arg1 form 399:402 a variant 74 residue non-glycosylated form 361:402 a variant 74 residue non-glycosylated form 361:402 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 8 gly non-glycosylated 382:397 arg1 Ser-CCL1 405:412 Ser-CCL1 405:412 Ser-CCL1 405:412 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 3 9 from effect 479:484 arg1 activities 521:530 the biological activities 506:530 the biological activities of either CCL1 or Ser-CCL1 506:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 3 10 gly glycosylation 489:501 arg1 Ser-CCL1 550:557 Ser-CCL1 550:557 Ser-CCL1 550:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 3 10 gly glycosylation 489:501 arg1 CCL1 542:545 CCL1 542:545 CCL1 542:545 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 3 11 from activities 521:530 arg1 studies 464:470 no systematic studies 450:470 no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1 450:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 6 12 theme non-glycosylated 939:954 arg1 proteins 956:963 the corresponding non-glycosylated proteins 921:963 the corresponding non-glycosylated proteins 921:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 3 13 theme CCL1 542:545 arg1 activities 521:530 the biological activities 506:530 the biological activities of either CCL1 or Ser-CCL1 506:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 7 14 theme molecules 1062:1070 arg1 structures 1029:1038 the chemical structures 1016:1038 the chemical structures of the (glyco)protein molecules 1016:1070 The results were correlated with the chemical structures of the (glyco)protein molecules. 24644239 4 15 theme CCL1 663:666 arg1 forms 648:652 both N-glycosylated and non-glycosylated forms 607:652 both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 607:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 3 16 theme systematic 453:462 arg1 studies 464:470 no systematic studies 450:470 no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1 450:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 8 17 gly N-glycosylated 1243:1256 arg1 molecules 1266:1274 homogeneous N-glycosylated protein molecules 1231:1274 homogeneous N-glycosylated protein molecules of defined covalent structure 1231:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 7 18 theme glyco 1048:1052 arg1 molecules 1062:1070 the (glyco)protein molecules 1043:1070 the (glyco)protein molecules 1043:1070 The results were correlated with the chemical structures of the (glyco)protein molecules. 24644239 5 19 theme phase 810:814 arg1 synthesis 816:824 Fmoc chemistry solid phase synthesis 789:824 Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 789:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 0 20 dep chemokines 101:110 arg1 chemokines 101:110 the chemokines CCL1 and Ser-CCL1 97:128 the chemokines CCL1 and Ser-CCL1 97:128 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 0 20 dep chemokines 101:110 arg1 Ser-CCL1 121:128 Ser-CCL1 121:128 Ser-CCL1 121:128 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 0 20 dep chemokines 101:110 arg1 CCL1 112:115 CCL1 112:115 CCL1 112:115 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 5 21 theme building 860:867 arg1 block 869:873 the glycopeptide-(α) thioester building block 829:873 the glycopeptide-(α) thioester building block 829:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 2 22 theme residue 374:380 arg1 form 399:402 a variant 74 residue non-glycosylated form 361:402 a variant 74 residue non-glycosylated form 361:402 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 22 theme residue 374:380 arg1 Ser-CCL1 405:412 Ser-CCL1 405:412 Ser-CCL1 405:412 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 6 23 theme corresponding 925:937 arg1 proteins 956:963 the corresponding non-glycosylated proteins 921:963 the corresponding non-glycosylated proteins 921:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 8 24 theme N-glycosylated 1243:1256 arg1 molecules 1266:1274 homogeneous N-glycosylated protein molecules 1231:1274 homogeneous N-glycosylated protein molecules of defined covalent structure 1231:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 25 from activity 1188:1195 arg1 investigations 1123:1136 the first investigations 1113:1136 the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1113:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 25 from activity 1188:1195 arg1 these 1103:1107 these 1103:1107 these 1103:1107 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 26 gly glycosylation 1155:1167 arg1 CCL1 1220:1223 the chemokine (Ser-)CCL1 1200:1223 the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1200:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 27 theme covalent 1287:1294 arg1 structure 1296:1304 defined covalent structure 1279:1304 defined covalent structure 1279:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 2 28 theme 74 371:372 arg1 residue 374:380 residue 374:380 residue 374:380 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 0 29 theme chemical 6:13 arg1 synthesis 15:23 Total chemical synthesis 0:23 Total chemical synthesis 0:23 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 2 30 theme amino 305:309 arg1 protein 316:322 a 73 amino acid protein 300:322 a 73 amino acid protein having one N-glycosylation site 300:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 30 theme amino 305:309 arg1 CCL1 277:280 CCL1 277:280 CCL1 277:280 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 31 theme variant 363:369 arg1 form 399:402 a variant 74 residue non-glycosylated form 361:402 a variant 74 residue non-glycosylated form 361:402 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 31 theme variant 363:369 arg1 Ser-CCL1 405:412 Ser-CCL1 405:412 Ser-CCL1 405:412 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 0 32 theme Total 0:4 arg1 synthesis 15:23 Total chemical synthesis 0:23 Total chemical synthesis 0:23 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 4 33 gly N-glycosylated 612:625 arg1 CCL1 663:666 (Ser-)CCL1 657:666 (Ser-)CCL1 657:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 4 33 gly N-glycosylated 612:625 arg1 forms 648:652 both N-glycosylated and non-glycosylated forms 607:652 both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 607:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 3 34 from studies 464:470 arg1 activities 521:530 the biological activities 506:530 the biological activities of either CCL1 or Ser-CCL1 506:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 4 35 theme chemical 690:697 arg1 ligation 699:706 convergent native chemical ligation 672:706 convergent native chemical ligation 672:706 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 4 36 theme forms 648:652 arg1 syntheses 594:602 the total chemical syntheses 575:602 the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 575:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 8 37 theme CCL1 1220:1223 arg1 activity 1188:1195 the chemotactic activity 1172:1195 the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1172:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 5 38 theme block 869:873 arg1 synthesis 816:824 Fmoc chemistry solid phase synthesis 789:824 Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 789:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 8 39 theme protein 1258:1264 arg1 molecules 1266:1274 homogeneous N-glycosylated protein molecules 1231:1274 homogeneous N-glycosylated protein molecules of defined covalent structure 1231:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 5 40 theme Nbz 774:776 arg1 linker 778:783 the Nbz linker 770:783 the Nbz linker 770:783 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 5 41 used used 712:715 arg2 We 709:710 We 709:710 We 709:710 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 8 42 theme structure 1296:1304 arg1 molecules 1266:1274 homogeneous N-glycosylated protein molecules 1231:1274 homogeneous N-glycosylated protein molecules of defined covalent structure 1231:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 3 43 theme effect 479:484 arg1 studies 464:470 no systematic studies 450:470 no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1 450:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 8 44 from investigations 1123:1136 arg1 activity 1188:1195 the chemotactic activity 1172:1195 the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1172:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 1 45 theme activated 202:210 arg1 T-cells 212:218 activated T-cells 202:218 activated T-cells 202:218 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 8 46 theme effect 1145:1150 arg1 investigations 1123:1136 the first investigations 1113:1136 the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1113:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 46 theme effect 1145:1150 arg1 these 1103:1107 these 1103:1107 these 1103:1107 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 47 theme Ser- 1215:1218 arg1 CCL1 1220:1223 the chemokine (Ser-)CCL1 1200:1223 the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1200:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 2 48 gly N-glycosylation 335:349 arg2 site 351:354 one N-glycosylation site 331:354 one N-glycosylation site 331:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 48 gly N-glycosylation 335:349 arg2 one 331:333 one 331:333 one 331:333 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 5 49 theme Fmoc 789:792 arg1 synthesis 816:824 Fmoc chemistry solid phase synthesis 789:824 Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 789:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 8 50 theme first 1117:1121 arg1 investigations 1123:1136 the first investigations 1113:1136 the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1113:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 50 theme first 1117:1121 arg1 these 1103:1107 these 1103:1107 these 1103:1107 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 8 51 theme chemotactic 1176:1186 arg1 activity 1188:1195 the chemotactic activity 1172:1195 the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1172:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 4 52 theme N-glycosylated 612:625 arg1 forms 648:652 both N-glycosylated and non-glycosylated forms 607:652 both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 607:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 0 53 theme biological 29:38 arg1 activities 40:49 biological activities 29:49 biological activities 29:49 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 5 54 theme hen 743:745 arg1 yolk 751:754 hen egg yolk 743:754 hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 743:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 5 55 theme chemistry 794:802 arg1 synthesis 816:824 Fmoc chemistry solid phase synthesis 789:824 Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 789:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 2 56 theme non-glycosylated 382:397 arg1 form 399:402 a variant 74 residue non-glycosylated form 361:402 a variant 74 residue non-glycosylated form 361:402 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 56 theme non-glycosylated 382:397 arg1 Ser-CCL1 405:412 Ser-CCL1 405:412 Ser-CCL1 405:412 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 0 57 theme glycosylated 54:65 arg1 synthesis 15:23 Total chemical synthesis 0:23 Total chemical synthesis 0:23 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 0 57 theme glycosylated 54:65 arg1 activities 40:49 biological activities 29:49 biological activities 29:49 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 8 58 theme defined 1279:1285 arg1 structure 1296:1304 defined covalent structure 1279:1304 defined covalent structure 1279:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 3 59 theme Ser-CCL1 550:557 arg1 activities 521:530 the biological activities 506:530 the biological activities of either CCL1 or Ser-CCL1 506:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 8 60 from effect 1145:1150 arg1 activity 1188:1195 the chemotactic activity 1172:1195 the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1172:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 7 61 theme chemical 1020:1027 arg1 structures 1029:1038 the chemical structures 1016:1038 the chemical structures of the (glyco)protein molecules 1016:1070 The results were correlated with the chemical structures of the (glyco)protein molecules. 24644239 6 62 theme Chemotaxis 876:885 arg1 assays 887:892 Chemotaxis assays 876:892 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins 876:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 2 63 theme N-glycosylation 335:349 arg1 site 351:354 one N-glycosylation site 331:354 one N-glycosylation site 331:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 5 64 attach isolated 729:736 arg2 N-glycan 720:727 an N-glycan 717:727 an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 717:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 5 64 attach isolated 729:736 arg1 yolk 751:754 hen egg yolk 743:754 hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 743:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 5 65 theme thioester 850:858 arg1 block 869:873 the glycopeptide-(α) thioester building block 829:873 the glycopeptide-(α) thioester building block 829:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 6 66 gly glycoproteins 903:915 arg1 glycoproteins 903:915 these glycoproteins 897:915 these glycoproteins 897:915 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 6 67 gly non-glycosylated 939:954 arg1 proteins 956:963 the corresponding non-glycosylated proteins 921:963 the corresponding non-glycosylated proteins 921:963 Chemotaxis assays of these glycoproteins and the corresponding non-glycosylated proteins were carried out. 24644239 4 68 theme native 683:688 arg1 ligation 699:706 convergent native chemical ligation 672:706 convergent native chemical ligation 672:706 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 8 69 theme chemokine 1204:1212 arg1 CCL1 1220:1223 the chemokine (Ser-)CCL1 1200:1223 the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1200:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 1 70 gly glycosylated 151:162 arg1 CCL1 131:134 CCL1 131:134 CCL1 131:134 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 70 gly glycosylated 151:162 arg1 protein 174:180 a naturally glycosylated chemokine protein 139:180 a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes 139:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 1 70 gly glycosylated 151:162 arg1 chemoattractant 234:248 a chemoattractant 232:248 a chemoattractant for monocytes 232:262 CCL1 is a naturally glycosylated chemokine protein that is secreted by activated T-cells and acts as a chemoattractant for monocytes. 24644239 3 71 theme biological 510:519 arg1 activities 521:530 the biological activities 506:530 the biological activities of either CCL1 or Ser-CCL1 506:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 5 72 theme solid 804:808 arg1 synthesis 816:824 Fmoc chemistry solid phase synthesis 789:824 Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 789:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 2 73 theme acid 311:314 arg1 protein 316:322 a 73 amino acid protein 300:322 a 73 amino acid protein having one N-glycosylation site 300:354 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 2 73 theme acid 311:314 arg1 CCL1 277:280 CCL1 277:280 CCL1 277:280 Originally, CCL1 was identified as a 73 amino acid protein having one N-glycosylation site, and a variant 74 residue non-glycosylated form, Ser-CCL1, has also been described. 24644239 8 74 theme glycosylation 1155:1167 arg1 effect 1145:1150 the effect 1141:1150 the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure 1141:1304 To the best of our knowledge, these are the first investigations of the effect of glycosylation on the chemotactic activity of the chemokine (Ser-)CCL1 using homogeneous N-glycosylated protein molecules of defined covalent structure. 24644239 4 75 theme convergent 672:681 arg1 ligation 699:706 convergent native chemical ligation 672:706 convergent native chemical ligation 672:706 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 0 76 theme non-glycosylated 71:86 arg1 synthesis 15:23 Total chemical synthesis 0:23 Total chemical synthesis 0:23 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 0 76 theme non-glycosylated 71:86 arg1 activities 40:49 biological activities 29:49 biological activities 29:49 Total chemical synthesis and biological activities of glycosylated and non-glycosylated forms of the chemokines CCL1 and Ser-CCL1. 24644239 5 77 theme glycopeptide- 833:845 arg1 block 869:873 the glycopeptide-(α) thioester building block 829:873 the glycopeptide-(α) thioester building block 829:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 4 78 theme Ser- 658:661 arg1 CCL1 663:666 (Ser-)CCL1 657:666 (Ser-)CCL1 657:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 4 79 theme non-glycosylated 631:646 arg1 forms 648:652 both N-glycosylated and non-glycosylated forms 607:652 both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 607:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 4 80 theme chemical 585:592 arg1 syntheses 594:602 the total chemical syntheses 575:602 the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 575:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 24644239 5 81 theme egg 747:749 arg1 yolk 751:754 hen egg yolk 743:754 hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block 743:873 We used an N-glycan isolated from hen egg yolk together with the Nbz linker for Fmoc chemistry solid phase synthesis of the glycopeptide-(α) thioester building block. 24644239 3 82 theme glycosylation 489:501 arg1 effect 479:484 the effect 475:484 the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1 475:557 There are no systematic studies of the effect of glycosylation on the biological activities of either CCL1 or Ser-CCL1. 24644239 4 83 theme total 579:583 arg1 syntheses 594:602 the total chemical syntheses 575:602 the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1 575:666 Here we report the total chemical syntheses of both N-glycosylated and non-glycosylated forms of (Ser-)CCL1, by convergent native chemical ligation. 27581986 7 0 theme monoclonal 1260:1269 arg1 antibody 1271:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 14 1 theme viral 2549:2553 arg1 sequencing 2560:2569 viral deep sequencing 2549:2569 viral deep sequencing 2549:2569 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 11 2 dep IMPORTANCE 1928:1937 arg1 site 1965:1968 The conserved CD4 binding site 1939:1968 IMPORTANCE The conserved CD4 binding site on gp120 1928:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 8 3 attach present 1548:1554 arg2 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 3 attach present 1548:1554 arg1 strains 1576:1582 almost all HIV-1 strains 1559:1582 almost all HIV-1 strains 1559:1582 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 3 attach present 1548:1554 arg2 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 1 4 theme neutralizing 270:281 arg1 antibodies 283:292 broadly neutralizing antibodies 262:292 broadly neutralizing antibodies 262:292 All HIV-1-infected individuals develop strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies. 27581986 5 5 from CAP257 1047:1052 arg1 representative 977:990 an N276 glycan-dependent CD4 binding site antibody representative 926:990 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 5 from CAP257 1047:1052 arg1 CAP257-RH1 914:923 CAP257-RH1 914:923 CAP257-RH1 914:923 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 1 6 theme neutralizing 180:191 arg1 antibodies 193:202 strain-specific neutralizing antibodies 164:202 strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies 164:292 All HIV-1-infected individuals develop strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies. 27581986 9 7 theme V5 1746:1747 arg1 glycans 1749:1755 V5 glycans 1746:1755 V5 glycans 1746:1755 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 11 8 from events 2031:2036 arg1 elicitation 2045:2055 elicitation 2045:2055 elicitation 2045:2055 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 11 8 from events 2031:2036 arg1 maturation 2061:2070 maturation 2061:2070 maturation 2061:2070 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 4 9 theme tier 738:741 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 2 10 theme strain-specific 320:334 arg1 antibodies 336:345 strain-specific antibodies 320:345 strain-specific antibodies that overlap conserved sites of vulnerability 320:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 5 11 theme N276 929:932 arg1 site 963:966 N276 glycan-dependent CD4 binding site 929:966 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 11 12 theme HIV-1 2001:2005 arg1 design 2015:2020 HIV-1 vaccine design 2001:2020 HIV-1 vaccine design 2001:2020 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 4 13 theme site 686:689 arg1 response 691:698 The initial CD4 binding site response 662:698 The initial CD4 binding site response 662:698 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 9 14 theme Deep 1633:1636 arg1 sequencing 1638:1647 Deep sequencing 1633:1647 Deep sequencing of autologous CAP257 viruses 1633:1676 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 0 15 theme CD4 107:109 arg1 Site 119:122 the CD4 Binding Site 103:122 the CD4 Binding Site 103:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 15 16 theme glycan 2718:2723 arg1 holes 2725:2729 glycan holes 2718:2729 glycan holes 2718:2729 These data highlight how glycan holes can play a role in the elicitation of B-cell lineages targeting the CD4 binding site. 27581986 3 17 theme glycan-dependent 612:627 arg1 epitope 629:635 an N276 glycan-dependent epitope 604:635 an N276 glycan-dependent epitope in the CD4 binding site 604:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 5 18 theme structural 883:892 arg1 characterization 894:909 structural characterization 883:909 structural characterization 883:909 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 11 19 theme lineages 2094:2101 arg1 elicitation 2045:2055 elicitation 2045:2055 elicitation 2045:2055 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 11 19 theme lineages 2094:2101 arg1 maturation 2061:2070 maturation 2061:2070 maturation 2061:2070 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 11 20 theme CD4 1953:1955 arg1 site 1965:1968 The conserved CD4 binding site 1939:1968 IMPORTANCE The conserved CD4 binding site on gp120 1928:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 12 21 theme strain-specific 2156:2170 arg1 antibodies 2172:2181 strain-specific antibodies 2156:2181 strain-specific antibodies 2156:2181 Studies have shown that strain-specific antibodies can evolve into broadly neutralizing antibodies or in some cases act as helper lineages. 27581986 15 22 theme CD4 2799:2801 arg1 site 2811:2814 the CD4 binding site 2795:2814 the CD4 binding site 2795:2814 These data highlight how glycan holes can play a role in the elicitation of B-cell lineages targeting the CD4 binding site. 27581986 13 23 theme HIV-1 2374:2378 arg1 immunogens 2380:2389 HIV-1 immunogens 2374:2389 HIV-1 immunogens 2374:2389 Therefore, characterizing the epitopes of strain-specific antibodies may help to inform the design of HIV-1 immunogens to elicit broadly neutralizing antibodies. 27581986 7 24 theme donor 1417:1421 arg1 CAP257 1423:1428 donor CAP257 1417:1428 donor CAP257 1417:1428 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 3 25 theme CD4 644:646 arg1 site 656:659 the CD4 binding site 640:659 the CD4 binding site 640:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 11 26 theme key 2027:2029 arg1 events 2031:2036 key events 2027:2036 key events in the elicitation and maturation of different antibody lineages to this site 2027:2114 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 10 27 theme CD4 1895:1897 arg1 site 1907:1910 N276 glycan-dependent CD4 binding site 1873:1910 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 6 28 theme CAP257-RH1 1082:1091 arg1 structure 1069:1077 The cocrystal structure 1055:1077 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 1055:1111 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 8 29 theme V5 1539:1540 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 29 theme V5 1539:1540 arg1 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 11 30 theme major 1984:1988 arg1 IMPORTANCE 1928:1937 IMPORTANCE 1928:1937 IMPORTANCE The conserved CD4 binding site on gp120 1928:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 11 30 theme major 1984:1988 arg1 target 1990:1995 a major target 1982:1995 a major target for HIV-1 vaccine design 1982:2020 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 4 31 theme clade 745:749 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 7 32 theme CAP257-RH1 1249:1258 arg1 antibody 1271:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 10 33 theme N276 1873:1876 arg1 site 1907:1910 N276 glycan-dependent CD4 binding site 1873:1910 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 5 34 theme CD4 951:953 arg1 site 963:966 N276 glycan-dependent CD4 binding site 929:966 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 14 35 theme glycan-dependent 2647:2662 arg1 site 2676:2679 glycan-dependent CD4 binding site 2647:2679 these early glycan-dependent CD4 binding site antibodies 2635:2690 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 8 36 theme present 1548:1554 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 36 theme present 1548:1554 arg1 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 4 37 theme viral 753:757 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 0 38 theme V5 76:77 arg1 Hole 86:89 a Rare V5 Glycan Hole 69:89 a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site 69:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 5 39 theme site 963:966 arg1 representative 977:990 an N276 glycan-dependent CD4 binding site antibody representative 926:990 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 39 theme site 963:966 arg1 CAP257-RH1 914:923 CAP257-RH1 914:923 CAP257-RH1 914:923 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 4 40 theme gp120 808:812 arg1 antigens 814:821 resurfaced gp120 antigens 797:821 resurfaced gp120 antigens for single-B-cell sorting 797:847 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 5 41 theme response 1029:1036 arg1 representative 977:990 an N276 glycan-dependent CD4 binding site antibody representative 926:990 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 41 theme response 1029:1036 arg1 CAP257-RH1 914:923 CAP257-RH1 914:923 CAP257-RH1 914:923 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 8 42 theme HIV-1 1570:1574 arg1 strains 1576:1582 almost all HIV-1 strains 1559:1582 almost all HIV-1 strains 1559:1582 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 5 43 theme donor 1041:1045 arg1 CAP257 1047:1052 donor CAP257 1041:1052 donor CAP257 1041:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 12 44 theme helper 2255:2260 arg1 lineages 2262:2269 helper lineages 2255:2269 helper lineages 2255:2269 Studies have shown that strain-specific antibodies can evolve into broadly neutralizing antibodies or in some cases act as helper lineages. 27581986 11 45 theme binding 1957:1963 arg1 site 1965:1968 The conserved CD4 binding site 1939:1968 IMPORTANCE The conserved CD4 binding site on gp120 1928:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 5 46 theme CD4 1005:1007 arg1 site 1017:1020 the early CD4 binding site 995:1020 the early CD4 binding site plasma response in donor CAP257 995:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 6 47 theme critical 1122:1129 arg1 interactions 1131:1142 critical interactions 1122:1142 critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, 1122:1212 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 3 48 theme clade 509:513 arg1 CAP257 533:538 CAP257 533:538 CAP257 533:538 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 3 48 theme clade 509:513 arg1 donor 526:530 an HIV-1 clade C-infected donor 500:530 an HIV-1 clade C-infected donor 500:530 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 8 49 from present 1548:1554 arg1 strains 1576:1582 almost all HIV-1 strains 1559:1582 almost all HIV-1 strains 1559:1582 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 6 50 with interactions 1131:1142 arg1 acid 1204:1207 aspartic acid 368 1195:1211 aspartic acid 368 1195:1211 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 6 50 with interactions 1131:1142 arg1 glycan 1158:1163 the N276 glycan 1149:1163 the N276 glycan 1149:1163 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 5 51 theme site 1017:1020 arg1 response 1029:1036 the early CD4 binding site plasma response 995:1036 the early CD4 binding site plasma response in donor CAP257 995:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 8 52 theme transmitted/founder 1606:1624 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 53 theme neutralization 1442:1455 arg1 breadth 1457:1463 Its narrow neutralization breadth 1431:1463 Its narrow neutralization breadth 1431:1463 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 2 54 theme neutralizing 445:456 arg1 antibodies 458:467 broadly neutralizing antibodies 437:467 broadly neutralizing antibodies 437:467 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 5 55 theme CAP257-RH1 914:923 arg1 characterization 894:909 structural characterization 883:909 structural characterization 883:909 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 55 theme CAP257-RH1 914:923 arg1 isolation 869:877 isolation 869:877 isolation 869:877 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 0 56 theme HIV-1 31:35 arg1 Antibody 50:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 9 57 theme CAP257 1663:1668 arg1 viruses 1670:1676 autologous CAP257 viruses 1652:1676 autologous CAP257 viruses 1652:1676 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 7 58 theme IGLV3-10 1338:1345 arg1 genes 1347:1351 IGLV3-10 genes 1338:1351 IGLV3-10 genes 1338:1351 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 10 59 theme site 1907:1910 arg1 lineage 1919:1925 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 8 60 with incompatible 1508:1519 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 60 with incompatible 1508:1519 arg1 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 0 61 theme Antibody 50:57 arg1 Structure 0:8 Structure 0:8 Structure of an N276-Dependent HIV-1 Neutralizing Antibody 0:57 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 12 62 theme neutralizing 2207:2218 arg1 antibodies 2220:2229 broadly neutralizing antibodies 2199:2229 broadly neutralizing antibodies 2199:2229 Studies have shown that strain-specific antibodies can evolve into broadly neutralizing antibodies or in some cases act as helper lineages. 27581986 14 63 theme N276 2484:2487 arg1 antibody 2506:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 11 64 theme vaccine 2007:2013 arg1 design 2015:2020 HIV-1 vaccine design 2001:2020 HIV-1 vaccine design 2001:2020 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 12 65 dep evolve 2187:2192 arg1 act 2248:2250 act 2248:2250 act as helper lineages 2248:2269 Studies have shown that strain-specific antibodies can evolve into broadly neutralizing antibodies or in some cases act as helper lineages. 27581986 8 66 gly glycosylated 1526:1537 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 66 gly glycosylated 1526:1537 arg1 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 1 67 theme HIV-1-infected 129:142 arg1 individuals 144:154 All HIV-1-infected individuals 125:154 All HIV-1-infected individuals 125:154 All HIV-1-infected individuals develop strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies. 27581986 8 68 theme binding 1485:1491 arg1 angle 1493:1497 a binding angle 1483:1497 a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus 1483:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 68 theme binding 1485:1491 arg1 incompatible 1508:1519 incompatible 1508:1519 incompatible 1508:1519 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 11 69 theme conserved 1943:1951 arg1 site 1965:1968 The conserved CD4 binding site 1939:1968 IMPORTANCE The conserved CD4 binding site on gp120 1928:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 15 70 theme B-cell 2769:2774 arg1 lineages 2776:2783 B-cell lineages 2769:2783 B-cell lineages targeting the CD4 binding site 2769:2814 These data highlight how glycan holes can play a role in the elicitation of B-cell lineages targeting the CD4 binding site. 27581986 9 71 theme minority 1697:1704 arg1 variants 1706:1713 minority variants 1697:1713 minority variants 1697:1713 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 4 72 theme initial 666:672 arg1 site 686:689 The initial CD4 binding site 662:689 The initial CD4 binding site response 662:698 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 2 73 theme vulnerability 379:391 arg1 vulnerability 379:391 vulnerability 379:391 vulnerability 379:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 2 73 theme vulnerability 379:391 arg1 sites 370:374 conserved sites 360:374 conserved sites of vulnerability 360:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 14 74 theme deep 2555:2558 arg1 sequencing 2560:2569 viral deep sequencing 2549:2569 viral deep sequencing 2549:2569 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 5 75 dep isolation 869:877 arg1 the 865:867 the 865:867 the 865:867 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 6 76 theme N276 1153:1156 arg1 glycan 1158:1163 the N276 glycan 1149:1163 the N276 glycan 1149:1163 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 4 77 used used 782:785 arg2 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 11 78 from site 1965:1968 arg1 gp120 1973:1977 gp120 1973:1977 gp120 1973:1977 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 11 79 theme different 2075:2083 arg1 lineages 2094:2101 different antibody lineages 2075:2101 different antibody lineages 2075:2101 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 14 80 theme binding 2668:2674 arg1 site 2676:2679 glycan-dependent CD4 binding site 2647:2679 these early glycan-dependent CD4 binding site antibodies 2635:2690 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 13 81 theme strain-specific 2314:2328 arg1 antibodies 2330:2339 strain-specific antibodies 2314:2339 strain-specific antibodies 2314:2339 Therefore, characterizing the epitopes of strain-specific antibodies may help to inform the design of HIV-1 immunogens to elicit broadly neutralizing antibodies. 27581986 10 82 theme V5 1776:1777 arg1 loops 1779:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 10 82 theme V5 1776:1777 arg1 holes 1797:1801 unusual holes 1789:1801 unusual holes 1789:1801 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 3 83 theme plasma 576:581 arg1 antibodies 583:592 broadly neutralizing plasma antibodies 555:592 broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site 555:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 1 84 theme strain-specific 164:178 arg1 antibodies 193:202 strain-specific neutralizing antibodies 164:202 strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies 164:292 All HIV-1-infected individuals develop strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies. 27581986 15 85 theme lineages 2776:2783 arg1 elicitation 2754:2764 the elicitation 2750:2764 the elicitation of B-cell lineages targeting the CD4 binding site 2750:2814 These data highlight how glycan holes can play a role in the elicitation of B-cell lineages targeting the CD4 binding site. 27581986 14 86 theme X-ray 2523:2527 arg1 crystallography 2529:2543 X-ray crystallography 2523:2543 X-ray crystallography 2523:2543 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 14 87 theme early 2641:2645 arg1 antibodies 2681:2690 these early glycan-dependent CD4 binding site antibodies 2635:2690 these early glycan-dependent CD4 binding site antibodies 2635:2690 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 6 88 theme aspartic 1195:1202 arg1 acid 1204:1207 aspartic acid 368 1195:1211 aspartic acid 368 1195:1211 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 13 89 theme immunogens 2380:2389 arg1 design 2364:2369 the design 2360:2369 the design of HIV-1 immunogens to elicit broadly neutralizing antibodies 2360:2431 Therefore, characterizing the epitopes of strain-specific antibodies may help to inform the design of HIV-1 immunogens to elicit broadly neutralizing antibodies. 27581986 2 90 theme antibodies 336:345 arg1 epitopes 308:315 the epitopes 304:315 the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability 304:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 10 91 from holes 1797:1801 arg1 necessary 1843:1851 necessary 1843:1851 necessary 1843:1851 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 10 91 from holes 1797:1801 arg1 shield 1817:1822 shield 1817:1822 shield 1817:1822 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 4 92 theme heterologous 725:736 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 0 93 theme Binding 111:117 arg1 Site 119:122 the CD4 Binding Site 103:122 the CD4 Binding Site 103:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 3 94 theme N276 607:610 arg1 epitope 629:635 an N276 glycan-dependent epitope 604:635 an N276 glycan-dependent epitope in the CD4 binding site 604:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 11 95 theme antibody 2085:2092 arg1 lineages 2094:2101 different antibody lineages 2075:2101 different antibody lineages 2075:2101 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 13 96 theme antibodies 2330:2339 arg1 epitopes 2302:2309 the epitopes 2298:2309 the epitopes of strain-specific antibodies 2298:2339 Therefore, characterizing the epitopes of strain-specific antibodies may help to inform the design of HIV-1 immunogens to elicit broadly neutralizing antibodies. 27581986 4 97 theme binding 678:684 arg1 site 686:689 The initial CD4 binding site 662:689 The initial CD4 binding site response 662:698 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 10 98 theme glycan-free 1764:1774 arg1 loops 1779:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 10 98 theme glycan-free 1764:1774 arg1 holes 1797:1801 unusual holes 1789:1801 unusual holes 1789:1801 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 3 99 theme binding 648:654 arg1 site 656:659 the CD4 binding site 640:659 the CD4 binding site 640:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 15 100 theme binding 2803:2809 arg1 site 2811:2814 the CD4 binding site 2795:2814 the CD4 binding site 2795:2814 These data highlight how glycan holes can play a role in the elicitation of B-cell lineages targeting the CD4 binding site. 27581986 5 101 theme antibody 968:975 arg1 representative 977:990 an N276 glycan-dependent CD4 binding site antibody representative 926:990 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 101 theme antibody 968:975 arg1 CAP257-RH1 914:923 CAP257-RH1 914:923 CAP257-RH1 914:923 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 8 102 theme glycosylated 1526:1537 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 8 102 theme glycosylated 1526:1537 arg1 loops 1542:1546 glycosylated V5 loops 1526:1546 glycosylated V5 loops 1526:1546 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 4 103 theme strain 759:764 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 10 104 theme binding 1899:1905 arg1 site 1907:1910 N276 glycan-dependent CD4 binding site 1873:1910 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 11 105 dep elicitation 2045:2055 arg1 the 2041:2043 the 2041:2043 the 2041:2043 IMPORTANCE The conserved CD4 binding site on gp120 is a major target for HIV-1 vaccine design, but key events in the elicitation and maturation of different antibody lineages to this site remain elusive. 27581986 14 106 theme site 2676:2679 arg1 antibodies 2681:2690 these early glycan-dependent CD4 binding site antibodies 2635:2690 these early glycan-dependent CD4 binding site antibodies 2635:2690 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 3 107 from epitope 629:635 arg1 site 656:659 the CD4 binding site 640:659 the CD4 binding site 640:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 0 108 theme Rare 71:74 arg1 Hole 86:89 a Rare V5 Glycan Hole 69:89 a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site 69:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 10 109 theme glycan-dependent 1878:1893 arg1 site 1907:1910 N276 glycan-dependent CD4 binding site 1873:1910 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 5 110 theme glycan-dependent 934:949 arg1 site 963:966 N276 glycan-dependent CD4 binding site 929:966 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 14 111 theme CD4 2664:2666 arg1 site 2676:2679 glycan-dependent CD4 binding site 2647:2679 these early glycan-dependent CD4 binding site antibodies 2635:2690 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 7 112 theme transmitted/founder 1386:1404 arg1 virus 1406:1410 the transmitted/founder virus 1382:1410 RHPA but not the transmitted/founder virus from donor CAP257 1369:1428 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 0 113 theme Glycan 79:84 arg1 Hole 86:89 a Rare V5 Glycan Hole 69:89 a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site 69:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 5 114 theme binding 955:961 arg1 site 963:966 N276 glycan-dependent CD4 binding site 929:966 an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257 926:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 6 115 theme cocrystal 1059:1067 arg1 structure 1069:1077 The cocrystal structure 1055:1077 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 1055:1111 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 8 116 theme all 1566:1568 arg1 strains 1576:1582 almost all HIV-1 strains 1559:1582 almost all HIV-1 strains 1559:1582 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 4 117 theme B 751:751 arg1 RHPA 766:769 the heterologous tier 2 clade B viral strain RHPA 721:769 the heterologous tier 2 clade B viral strain RHPA 721:769 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 0 118 theme Adjacent 91:98 arg1 Hole 86:89 a Rare V5 Glycan Hole 69:89 a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site 69:122 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 5 119 theme plasma 1022:1027 arg1 response 1029:1036 the early CD4 binding site plasma response 995:1036 the early CD4 binding site plasma response in donor CAP257 995:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 4 120 theme resurfaced 797:806 arg1 antigens 814:821 resurfaced gp120 antigens 797:821 resurfaced gp120 antigens for single-B-cell sorting 797:847 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 1 121 theme infecting 213:221 arg1 virus 223:227 their infecting virus 207:227 their infecting virus 207:227 All HIV-1-infected individuals develop strain-specific neutralizing antibodies to their infecting virus, which in some cases mature into broadly neutralizing antibodies. 27581986 5 122 theme early 999:1003 arg1 site 1017:1020 the early CD4 binding site 995:1020 the early CD4 binding site plasma response in donor CAP257 995:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 8 123 theme CAP257 1599:1604 arg1 virus 1626:1630 the CAP257 transmitted/founder virus 1595:1630 the CAP257 transmitted/founder virus 1595:1630 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 2 124 theme mechanistic 407:417 arg1 insights 419:426 mechanistic insights 407:426 mechanistic insights into how broadly neutralizing antibodies arise 407:473 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 0 125 theme N276-Dependent 16:29 arg1 Antibody 50:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 5 126 from representative 977:990 arg1 CAP257 1047:1052 donor CAP257 1041:1052 donor CAP257 1041:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 5 127 theme binding 1009:1015 arg1 site 1017:1020 the early CD4 binding site 995:1020 the early CD4 binding site plasma response in donor CAP257 995:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 7 128 from CAP257 1423:1428 arg1 virus 1406:1410 the transmitted/founder virus 1382:1410 RHPA but not the transmitted/founder virus from donor CAP257 1369:1428 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 3 129 theme C-infected 515:524 arg1 CAP257 533:538 CAP257 533:538 CAP257 533:538 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 3 129 theme C-infected 515:524 arg1 donor 526:530 an HIV-1 clade C-infected donor 500:530 an HIV-1 clade C-infected donor 500:530 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 0 130 theme Neutralizing 37:48 arg1 Antibody 50:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 an N276-Dependent HIV-1 Neutralizing Antibody 13:57 Structure of an N276-Dependent HIV-1 Neutralizing Antibody Targeting a Rare V5 Glycan Hole Adjacent to the CD4 Binding Site. 27581986 5 131 from response 1029:1036 arg1 CAP257 1047:1052 donor CAP257 1041:1052 donor CAP257 1041:1052 Here we report the isolation and structural characterization of CAP257-RH1, an N276 glycan-dependent CD4 binding site antibody representative of the early CD4 binding site plasma response in donor CAP257. 27581986 9 132 theme autologous 1652:1661 arg1 viruses 1670:1676 autologous CAP257 viruses 1652:1676 autologous CAP257 viruses 1652:1676 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 6 133 theme RHPA 1102:1105 arg1 gp120 1107:1111 RHPA gp120 1102:1111 RHPA gp120 1102:1111 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 10 134 theme B-cell 1912:1917 arg1 lineage 1919:1925 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 this N276 glycan-dependent CD4 binding site B-cell lineage 1868:1925 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 8 135 theme narrow 1435:1440 arg1 breadth 1457:1463 Its narrow neutralization breadth 1431:1463 Its narrow neutralization breadth 1431:1463 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 3 136 theme HIV-1 503:507 arg1 CAP257 533:538 CAP257 533:538 CAP257 533:538 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 3 136 theme HIV-1 503:507 arg1 donor 526:530 an HIV-1 clade C-infected donor 500:530 an HIV-1 clade C-infected donor 500:530 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 27581986 8 137 from strains 1576:1582 arg1 present 1548:1554 present 1548:1554 present 1548:1554 Its narrow neutralization breadth was attributed to a binding angle that was incompatible with glycosylated V5 loops present in almost all HIV-1 strains, including the CAP257 transmitted/founder virus. 27581986 9 138 theme viruses 1670:1676 arg1 sequencing 1638:1647 Deep sequencing 1633:1647 Deep sequencing of autologous CAP257 viruses 1633:1676 Deep sequencing of autologous CAP257 viruses, however, revealed minority variants early in infection that lacked V5 glycans. 27581986 7 139 theme immunoglobulin-variable 1301:1323 arg1 IGHV3-33 1325:1332 the immunoglobulin-variable IGHV3-33 1297:1332 the immunoglobulin-variable IGHV3-33 1297:1332 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 14 140 theme glycan-dependent 2489:2504 arg1 antibody 2506:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 2 141 theme conserved 360:368 arg1 vulnerability 379:391 vulnerability 379:391 vulnerability 379:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 2 141 theme conserved 360:368 arg1 sites 370:374 conserved sites 360:374 conserved sites of vulnerability 360:391 Defining the epitopes of strain-specific antibodies that overlap conserved sites of vulnerability might provide mechanistic insights into how broadly neutralizing antibodies arise. 27581986 13 142 theme neutralizing 2409:2420 arg1 antibodies 2422:2431 broadly neutralizing antibodies 2401:2431 broadly neutralizing antibodies 2401:2431 Therefore, characterizing the epitopes of strain-specific antibodies may help to inform the design of HIV-1 immunogens to elicit broadly neutralizing antibodies. 27581986 4 143 theme single-B-cell 827:839 arg1 sorting 841:847 single-B-cell sorting 827:847 single-B-cell sorting 827:847 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 14 144 theme neutralizing 2471:2482 arg1 antibody 2506:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 a narrowly neutralizing N276 glycan-dependent antibody 2460:2513 In this study, we isolate a narrowly neutralizing N276 glycan-dependent antibody and use X-ray crystallography and viral deep sequencing to describe how gp120 lacking glycans in V5 might have elicited these early glycan-dependent CD4 binding site antibodies. 27581986 10 145 theme unusual 1789:1795 arg1 loops 1779:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops 1758:1783 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 10 145 theme unusual 1789:1795 arg1 holes 1797:1801 unusual holes 1789:1801 unusual holes 1789:1801 These glycan-free V5 loops are unusual holes in the glycan shield that may have been necessary for initiating this N276 glycan-dependent CD4 binding site B-cell lineage. 27581986 6 146 theme loop 1166:1169 arg1 D 1171:1171 loop D 1166:1171 loop D 1166:1171 The cocrystal structure of CAP257-RH1 bound to RHPA gp120 revealed critical interactions with the N276 glycan, loop D, and V5, but not with aspartic acid 368, similarly to HJ16 and 179NC75. 27581986 4 147 theme CD4 674:676 arg1 site 686:689 The initial CD4 binding site 662:689 The initial CD4 binding site response 662:698 The initial CD4 binding site response potently neutralized the heterologous tier 2 clade B viral strain RHPA, which was used to design resurfaced gp120 antigens for single-B-cell sorting. 27581986 7 148 attach derived 1284:1290 arg1 genes 1347:1351 IGLV3-10 genes 1338:1351 IGLV3-10 genes 1338:1351 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 7 148 attach derived 1284:1290 arg2 antibody 1271:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody 1245:1278 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 7 148 attach derived 1284:1290 arg1 IGHV3-33 1325:1332 the immunoglobulin-variable IGHV3-33 1297:1332 the immunoglobulin-variable IGHV3-33 1297:1332 The CAP257-RH1 monoclonal antibody was derived from the immunoglobulin-variable IGHV3-33 and IGLV3-10 genes and neutralized RHPA but not the transmitted/founder virus from donor CAP257. 27581986 3 149 theme neutralizing 563:574 arg1 antibodies 583:592 broadly neutralizing plasma antibodies 555:592 broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site 555:659 We previously described an HIV-1 clade C-infected donor, CAP257, who developed broadly neutralizing plasma antibodies targeting an N276 glycan-dependent epitope in the CD4 binding site. 29279989 8 0 theme composite 1146:1154 arg1 design 1156:1161 a central composite design 1136:1161 a central composite design for derivatization of O-linked glycans 1136:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 3 1 theme O-linked 513:520 arg1 glycans 522:528 O-linked glycans 513:528 O-linked glycans 513:528 We have taken the INLIGHT™ strategy and adopted it for use with O-linked glycans. 29279989 1 2 link N-linked 162:169 arg1 derivatization 178:191 N-linked glycan derivatization 162:191 N-linked glycan derivatization 162:191 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 6 3 theme removed 907:913 arg1 glycans 915:921 the removed glycans 903:921 the removed glycans 903:921 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 9 4 theme optimized 1291:1299 arg1 conditions 1301:1310 the optimized conditions 1287:1310 the optimized conditions 1287:1310 Two glycoprotein standards were deglycosylated through hydrazinolysis and tagged at the optimized conditions. 29279989 2 5 theme surface 358:364 arg1 area 366:369 their non-polar surface area 342:369 their non-polar surface area 342:369 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 1 6 theme glycan 259:264 arg1 analysis 266:273 glycan analysis 259:273 glycan analysis 259:273 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 0 7 theme optimization 95:106 arg1 design 73:78 a central composite design 53:78 a central composite design of experiments optimization using the INLIGHT™ reagent 53:133 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 0 7 theme optimization 95:106 arg1 O-glycans 39:47 O-glycans 39:47 O-glycans 39:47 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 6 8 theme immunoglobulin 792:805 arg1 IgA 810:812 IgA 810:812 IgA 810:812 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 8 theme immunoglobulin 792:805 arg1 A 807:807 Human immunoglobulin A 786:807 Human immunoglobulin A (IgA) 786:813 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 4 9 used utilized 562:569 arg2 design 551:556 A central composite design 531:556 A central composite design 531:556 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 7 10 theme tagged 968:973 arg1 glycans 975:981 tagged glycans 968:981 tagged glycans 968:981 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 0 11 theme experiments 83:93 arg1 optimization 95:106 experiments optimization 83:106 experiments optimization using the INLIGHT™ reagent 83:133 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 5 12 from conditions 698:707 arg1 Derivatization 670:683 Derivatization 670:683 Derivatization at optimized conditions 670:707 Derivatization at optimized conditions was much quicker than any hydrazide derivatization strategy used previously. 29279989 4 13 dep find 574:577 arg1 % 611:611 45% 609:611 45% acetic acid 609:623 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 7 14 theme XIC 961:963 arg1 data 991:994 XIC of tagged glycans and MS2 data 961:994 XIC of tagged glycans and MS2 data 961:994 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 2 15 theme hydrazide 280:288 arg1 tag 290:292 The hydrazide tag 276:292 The hydrazide tag 276:292 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 1 16 theme glycan 171:176 arg1 derivatization 178:191 N-linked glycan derivatization 162:191 N-linked glycan derivatization 162:191 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 5 17 theme hydrazide 735:743 arg1 strategy 760:767 any hydrazide derivatization strategy 731:767 any hydrazide derivatization strategy used previously 731:783 Derivatization at optimized conditions was much quicker than any hydrazide derivatization strategy used previously. 29279989 9 18 theme glycoprotein 1207:1218 arg1 standards 1220:1228 Two glycoprotein standards 1203:1228 Two glycoprotein standards 1203:1228 Two glycoprotein standards were deglycosylated through hydrazinolysis and tagged at the optimized conditions. 29279989 10 19 theme MS/MS 1313:1317 arg1 data 1319:1322 MS/MS data 1313:1322 MS/MS data 1313:1322 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 8 20 theme glycans 1194:1200 arg1 derivatization 1167:1180 derivatization 1167:1180 derivatization of O-linked glycans 1167:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 8 21 theme central 1138:1144 arg1 design 1156:1161 a central composite design 1136:1161 a central composite design for derivatization of O-linked glycans 1136:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 3 22 with use 504:506 arg1 glycans 522:528 O-linked glycans 513:528 O-linked glycans 513:528 We have taken the INLIGHT™ strategy and adopted it for use with O-linked glycans. 29279989 5 23 theme derivatization 745:758 arg1 strategy 760:767 any hydrazide derivatization strategy 731:767 any hydrazide derivatization strategy used previously 731:783 Derivatization at optimized conditions was much quicker than any hydrazide derivatization strategy used previously. 29279989 8 24 theme hydrazide 1102:1110 arg1 tag 1112:1114 The INLIGHT™ hydrazide tag 1089:1114 The INLIGHT™ hydrazide tag 1089:1114 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 2 25 theme reversed-phase 385:398 arg1 separations 400:410 reversed-phase separations 385:410 reversed-phase separations 385:410 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 8 26 theme O-linked 1185:1192 arg1 glycans 1194:1200 O-linked glycans 1185:1200 O-linked glycans 1185:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 6 27 theme bovine 819:824 arg1 mucin 839:843 bovine submaxillary mucin 819:843 bovine submaxillary mucin (BSM) 819:849 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 27 theme bovine 819:824 arg1 BSM 846:848 BSM 846:848 BSM 846:848 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 28 theme Human 786:790 arg1 IgA 810:812 IgA 810:812 IgA 810:812 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 28 theme Human 786:790 arg1 A 807:807 Human immunoglobulin A 786:807 Human immunoglobulin A (IgA) 786:813 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 10 29 theme glycans 1357:1363 arg1 derivatization 1339:1352 INLIGHT™ derivatization 1330:1352 INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time 1330:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 0 30 theme tagging 27:33 arg1 Demonstration 0:12 Demonstration 0:12 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 0:134 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 3 31 link O-linked 513:520 arg1 glycans 522:528 O-linked glycans 513:528 O-linked glycans 513:528 We have taken the INLIGHT™ strategy and adopted it for use with O-linked glycans. 29279989 8 32 link O-linked 1185:1192 arg1 glycans 1194:1200 O-linked glycans 1185:1200 O-linked glycans 1185:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 4 33 theme tag 636:638 arg1 concentration 640:652 0.1 μg/μL tag concentration 626:652 0.1 μg/μL tag concentration 626:652 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 4 33 theme tag 636:638 arg1 C 658:658 37 C 655:658 37 C 655:658 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 0 34 theme hydrazide 17:25 arg1 tagging 27:33 hydrazide tagging 17:33 hydrazide tagging 17:33 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 4 35 theme optimized 579:587 arg1 conditions 597:606 optimized tagging conditions 579:606 optimized tagging conditions 579:606 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 0 36 theme INLIGHT™ 118:125 arg1 reagent 127:133 the INLIGHT™ reagent 114:133 the INLIGHT™ reagent 114:133 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 1 37 theme N-linked 162:169 arg1 derivatization 178:191 N-linked glycan derivatization 162:191 N-linked glycan derivatization 162:191 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 4 38 theme 0.1 μg/μL 626:634 arg1 concentration 640:652 0.1 μg/μL tag concentration 626:652 0.1 μg/μL tag concentration 626:652 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 4 38 theme 0.1 μg/μL 626:634 arg1 C 658:658 37 C 655:658 37 C 655:658 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 8 39 theme INLIGHT™ 1093:1100 arg1 tag 1112:1114 The INLIGHT™ hydrazide tag 1089:1114 The INLIGHT™ hydrazide tag 1089:1114 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 3 40 theme INLIGHT™ 467:474 arg1 strategy 476:483 the INLIGHT™ strategy 463:483 the INLIGHT™ strategy 463:483 We have taken the INLIGHT™ strategy and adopted it for use with O-linked glycans. 29279989 8 41 theme Graphical 1070:1078 arg1 abstract 1080:1087 Graphical abstract 1070:1087 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 1070:1201 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 10 42 theme INLIGHT™ 1330:1337 arg1 derivatization 1339:1352 INLIGHT™ derivatization 1330:1352 INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time 1330:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 2 43 theme ionization 426:435 arg1 efficiency 437:446 increased ionization efficiency 416:446 increased ionization efficiency 416:446 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 9 44 gly glycoprotein 1207:1218 arg1 glycoprotein 1207:1218 Two glycoprotein standards 1203:1228 Two glycoprotein standards 1203:1228 Two glycoprotein standards were deglycosylated through hydrazinolysis and tagged at the optimized conditions. 29279989 4 45 theme composite 541:549 arg1 design 551:556 A central composite design 531:556 A central composite design 531:556 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 4 46 theme tagging 589:595 arg1 conditions 597:606 optimized tagging conditions 579:606 optimized tagging conditions 579:606 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 7 47 theme hydrazide 1012:1020 arg1 tagging 1022:1028 successful hydrazide tagging 1001:1028 successful hydrazide tagging of O-linked glycans for the first time 1001:1067 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 2 48 theme increased 416:424 arg1 efficiency 437:446 increased ionization efficiency 416:446 increased ionization efficiency 416:446 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 8 49 dep abstract 1080:1087 arg1 optimized 1120:1128 optimized 1120:1128 was optimized using a central composite design for derivatization of O-linked glycans 1116:1200 Graphical abstract The INLIGHT™ hydrazide tag was optimized using a central composite design for derivatization of O-linked glycans. 29279989 4 50 theme central 533:539 arg1 design 551:556 A central composite design 531:556 A central composite design 531:556 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 7 51 theme glycans 1042:1048 arg1 tagging 1022:1028 successful hydrazide tagging 1001:1028 successful hydrazide tagging of O-linked glycans for the first time 1001:1067 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 10 52 theme hydrazide 1390:1398 arg1 tagging 1400:1406 successful hydrazide tagging 1379:1406 successful hydrazide tagging of O-glycans for the first time 1379:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 2 53 theme non-polar 348:356 arg1 area 366:369 their non-polar surface area 342:369 their non-polar surface area 342:369 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 7 54 theme first 1058:1062 arg1 time 1064:1067 the first time 1054:1067 the first time 1054:1067 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 5 55 theme optimized 688:696 arg1 conditions 698:707 optimized conditions 688:707 optimized conditions 688:707 Derivatization at optimized conditions was much quicker than any hydrazide derivatization strategy used previously. 29279989 10 56 theme successful 1379:1388 arg1 tagging 1400:1406 successful hydrazide tagging 1379:1406 successful hydrazide tagging of O-glycans for the first time 1379:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 7 57 link O-linked 1033:1040 arg1 glycans 1042:1048 O-linked glycans 1033:1048 O-linked glycans 1033:1048 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 10 58 theme first 1429:1433 arg1 time 1435:1438 the first time 1425:1438 the first time 1425:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 7 59 theme successful 1001:1010 arg1 tagging 1022:1028 successful hydrazide tagging 1001:1028 successful hydrazide tagging of O-linked glycans for the first time 1001:1067 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 2 60 dep glycans 322:328 arg1 increasing 331:340 increasing 331:340 increasing their non-polar surface area 331:369 The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. 29279989 10 61 theme O-glycans 1411:1419 arg1 tagging 1400:1406 successful hydrazide tagging 1379:1406 successful hydrazide tagging of O-glycans for the first time 1379:1438 MS/MS data shows INLIGHT™ derivatization of glycans demonstrating successful hydrazide tagging of O-glycans for the first time. 29279989 4 62 theme acetic 613:618 arg1 acid 620:623 acetic acid 613:623 45% acetic acid 609:623 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 7 63 theme O-linked 1033:1040 arg1 glycans 1042:1048 O-linked glycans 1033:1048 O-linked glycans 1033:1048 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 6 64 theme submaxillary 826:837 arg1 mucin 839:843 bovine submaxillary mucin 819:843 bovine submaxillary mucin (BSM) 819:849 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 64 theme submaxillary 826:837 arg1 BSM 846:848 BSM 846:848 BSM 846:848 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 7 65 theme MS2 987:989 arg1 data 991:994 XIC of tagged glycans and MS2 data 961:994 XIC of tagged glycans and MS2 data 961:994 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 6 66 gly deglycosylated 861:874 arg1 mucin 839:843 bovine submaxillary mucin 819:843 bovine submaxillary mucin (BSM) 819:849 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 66 gly deglycosylated 861:874 arg1 IgA 810:812 IgA 810:812 IgA 810:812 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 66 gly deglycosylated 861:874 arg1 A 807:807 Human immunoglobulin A 786:807 Human immunoglobulin A (IgA) 786:813 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 6 66 gly deglycosylated 861:874 arg1 BSM 846:848 BSM 846:848 BSM 846:848 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29279989 0 67 theme composite 63:71 arg1 design 73:78 a central composite design 53:78 a central composite design of experiments optimization using the INLIGHT™ reagent 53:133 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 4 68 dep % 611:611 arg1 concentration 640:652 0.1 μg/μL tag concentration 626:652 0.1 μg/μL tag concentration 626:652 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 4 68 dep % 611:611 arg1 C 658:658 37 C 655:658 37 C 655:658 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 4 68 dep % 611:611 arg1 acid 620:623 acetic acid 613:623 45% acetic acid 609:623 A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). 29279989 9 69 gly deglycosylated 1235:1248 arg1 standards 1220:1228 Two glycoprotein standards 1203:1228 Two glycoprotein standards 1203:1228 Two glycoprotein standards were deglycosylated through hydrazinolysis and tagged at the optimized conditions. 29279989 1 70 theme INLIGHT™ 140:147 arg1 strategy 149:156 The INLIGHT™ strategy 136:156 The INLIGHT™ strategy for N-linked glycan derivatization 136:191 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 1 71 theme challenges 232:241 arg1 challenges 232:241 the challenges 228:241 the challenges associated with glycan analysis 228:273 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 1 71 theme challenges 232:241 arg1 many 220:223 many 220:223 many 220:223 The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. 29279989 0 72 theme central 55:61 arg1 design 73:78 a central composite design 53:78 a central composite design of experiments optimization using the INLIGHT™ reagent 53:133 Demonstration of hydrazide tagging for O-glycans and a central composite design of experiments optimization using the INLIGHT™ reagent. 29279989 7 73 theme glycans 975:981 arg1 MS2 987:989 MS2 987:989 MS2 987:989 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 7 73 theme glycans 975:981 arg1 XIC 961:963 XIC 961:963 XIC of tagged glycans 961:981 XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time. 29279989 6 74 theme optimum 941:947 arg1 conditions 949:958 optimum conditions 941:958 optimum conditions 941:958 Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. 29058712 2 0 theme synthetic 489:497 arg1 analog 503:508 a synthetic LLO analog 487:508 a synthetic LLO analog 487:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 6 1 theme heptasaccharide 872:886 arg1 LLO 888:890 full-size heptasaccharide LLO 862:890 full-size heptasaccharide LLO 862:890 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 2 2 theme single-subunit 371:384 arg1 PglB 391:394 a single-subunit OST, PglB 369:394 PglB 391:394 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 3 3 from role 537:540 arg1 recognition 609:619 substrate recognition 599:619 substrate recognition 599:619 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 7 4 theme functional 1025:1034 arg1 roles 1036:1040 functional roles 1025:1040 functional roles 1025:1040 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 6 5 theme full-size 862:870 arg1 LLO 888:890 full-size heptasaccharide LLO 862:890 full-size heptasaccharide LLO 862:890 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 0 6 link lipid-linked 19:30 arg1 oligosaccharide 32:46 lipid-linked oligosaccharide recognition and processing 19:73 oligosaccharide 32:46 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 6 7 theme EL5 808:810 arg1 mobility 812:819 EL5 mobility 808:819 EL5 mobility 808:819 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 2 8 theme X-ray 350:354 arg1 structure 356:364 the X-ray structure 346:364 the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog 346:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 3 9 attach present 568:574 arg2 EL5 563:565 the external loop EL5 545:565 the external loop EL5 545:565 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 3 9 attach present 568:574 arg1 enzymes 587:593 all OST enzymes 579:593 all OST enzymes 579:593 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 1 10 theme first 291:295 arg1 enzyme 170:175 a membrane-integral enzyme 150:175 a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains 150:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 1 10 theme first 291:295 arg1 step 297:300 the first step 287:300 the first step in protein N-glycosylation 287:327 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 4 11 theme acceptor 707:714 arg1 peptide 716:722 the acceptor peptide 703:722 the acceptor peptide 703:722 Whereas the N-terminal half of EL5 binds LLO, the C-terminal half interacts with the acceptor peptide. 29058712 6 12 theme OST 853:855 arg1 rate 845:848 the catalytic rate 831:848 the catalytic rate of OST 831:855 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 3 13 from enzymes 587:593 arg1 present 568:574 present 568:574 present 568:574 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 6 14 theme LLO 945:947 arg1 analog 949:954 a monosaccharide-containing LLO analog 917:954 a monosaccharide-containing LLO analog 917:954 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 0 15 theme processing 64:73 arg1 basis 10:14 Molecular basis 0:14 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 0:113 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 7 16 theme conserved 1045:1053 arg1 motifs 1059:1064 conserved OST motifs 1045:1064 conserved OST motifs 1045:1064 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 2 17 from structure 356:364 arg1 lari 415:418 lari 415:418 lari 415:418 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 1 18 link lipid-linked 221:232 arg1 LLOs 252:255 LLOs 252:255 LLOs 252:255 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 1 18 link lipid-linked 221:232 arg1 oligosaccharides 234:249 lipid-linked oligosaccharides 221:249 lipid-linked oligosaccharides (LLOs) 221:256 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 7 19 theme PglB 1136:1139 arg1 design 1126:1131 rational design 1117:1131 rational design of PglB 1117:1139 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 1 20 theme glycans 208:214 arg1 transfer 196:203 the transfer 192:203 the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains 192:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 6 21 theme catalytic 835:843 arg1 rate 845:848 the catalytic rate 831:848 the catalytic rate of OST 831:855 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 6 22 theme monosaccharide-containing 919:943 arg1 analog 949:954 a monosaccharide-containing LLO analog 917:954 a monosaccharide-containing LLO analog 917:954 Reducing EL5 mobility decreases the catalytic rate of OST when full-size heptasaccharide LLO is provided, but not for a monosaccharide-containing LLO analog. 29058712 3 23 theme EL5 563:565 arg1 role 537:540 the role 533:540 the role of the external loop EL5, present in all OST enzymes, in substrate recognition 533:619 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 4 24 theme EL5 653:655 arg1 half 645:648 the N-terminal half 630:648 the N-terminal half of EL5 630:655 Whereas the N-terminal half of EL5 binds LLO, the C-terminal half interacts with the acceptor peptide. 29058712 0 25 theme Molecular 0:8 arg1 basis 10:14 Molecular basis 0:14 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 0:113 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 3 26 theme loop 558:561 arg1 EL5 563:565 the external loop EL5 545:565 the external loop EL5 545:565 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 3 27 from present 568:574 arg1 enzymes 587:593 all OST enzymes 579:593 all OST enzymes 579:593 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 2 28 theme PglB 391:394 arg1 structure 356:364 the X-ray structure 346:364 the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog 346:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 0 29 theme lipid-linked 19:30 arg1 oligosaccharide 32:46 lipid-linked oligosaccharide recognition and processing 19:73 oligosaccharide 32:46 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 1 30 theme protein 305:311 arg1 N-glycosylation 313:327 protein N-glycosylation 305:327 protein N-glycosylation 305:327 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 3 31 theme external 549:556 arg1 EL5 563:565 the external loop EL5 545:565 the external loop EL5 545:565 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 7 32 theme OST 1055:1057 arg1 motifs 1059:1064 conserved OST motifs 1045:1064 conserved OST motifs 1045:1064 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 5 33 theme active 786:791 arg1 site 793:796 the active site 782:796 the active site 782:796 The glycan moiety of LLO must thread under EL5 to access the active site. 29058712 2 34 theme intermediate 435:446 arg1 state 448:452 an intermediate state 432:452 an intermediate state bound to an acceptor peptide and a synthetic LLO analog 432:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 1 35 from step 297:300 arg1 N-glycosylation 313:327 protein N-glycosylation 305:327 protein N-glycosylation 305:327 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 2 36 from lari 415:418 arg1 structure 356:364 the X-ray structure 346:364 the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog 346:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 2 36 from lari 415:418 arg1 PglB 391:394 a single-subunit OST, PglB 369:394 PglB 391:394 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 5 37 theme LLO 746:748 arg1 moiety 736:741 The glycan moiety 725:741 The glycan moiety of LLO 725:748 The glycan moiety of LLO must thread under EL5 to access the active site. 29058712 7 38 theme state 1011:1015 arg1 chemistry 980:988 the chemistry 976:988 the chemistry of a ternary complex state 976:1015 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 1 39 theme lipid-linked 221:232 arg1 LLOs 252:255 LLOs 252:255 LLOs 252:255 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 1 39 theme lipid-linked 221:232 arg1 oligosaccharides 234:249 lipid-linked oligosaccharides 221:249 lipid-linked oligosaccharides (LLOs) 221:256 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 0 40 theme oligosaccharide 32:46 arg1 basis 10:14 Molecular basis 0:14 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 0:113 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 3 41 theme OST 583:585 arg1 enzymes 587:593 all OST enzymes 579:593 all OST enzymes 579:593 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 2 42 theme acceptor 466:473 arg1 peptide 475:481 an acceptor peptide 463:481 an acceptor peptide 463:481 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 7 43 theme ternary 995:1001 arg1 complex 1003:1009 a ternary complex 993:1009 a ternary complex state 993:1015 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 5 44 theme glycan 729:734 arg1 moiety 736:741 The glycan moiety 725:741 The glycan moiety of LLO 725:748 The glycan moiety of LLO must thread under EL5 to access the active site. 29058712 3 45 theme present 568:574 arg1 EL5 563:565 the external loop EL5 545:565 the external loop EL5 545:565 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 7 46 theme complex 1003:1009 arg1 state 1011:1015 a ternary complex state 993:1015 a ternary complex state 993:1015 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 0 47 dep oligosaccharide 32:46 arg1 recognition 48:58 recognition 48:58 recognition 48:58 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 1 48 from oligosaccharides 234:249 arg1 transfer 196:203 the transfer 192:203 the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains 192:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 4 49 theme C-terminal 672:681 arg1 half 683:686 the C-terminal half 668:686 the C-terminal half 668:686 Whereas the N-terminal half of EL5 binds LLO, the C-terminal half interacts with the acceptor peptide. 29058712 7 50 theme rational 1117:1124 arg1 design 1126:1131 rational design 1117:1131 rational design of PglB 1117:1139 Our results define the chemistry of a ternary complex state, assign functional roles to conserved OST motifs, and provide opportunities for glycoengineering by rational design of PglB. 29058712 1 51 theme membrane-integral 152:168 arg1 enzyme 170:175 a membrane-integral enzyme 150:175 a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains 150:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 1 51 theme membrane-integral 152:168 arg1 Oligosaccharyltransferase 115:139 Oligosaccharyltransferase 115:139 Oligosaccharyltransferase (OST) 115:145 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 1 51 theme membrane-integral 152:168 arg1 step 297:300 the first step 287:300 the first step in protein N-glycosylation 287:327 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 3 52 theme substrate 599:607 arg1 recognition 609:619 substrate recognition 599:619 substrate recognition 599:619 The structure reveals the role of the external loop EL5, present in all OST enzymes, in substrate recognition. 29058712 1 53 theme asparagine 263:272 arg1 chains 279:284 asparagine side chains 263:284 asparagine side chains 263:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 0 54 theme bacterial 78:86 arg1 oligosaccharyltransferase 88:112 bacterial oligosaccharyltransferase 78:112 bacterial oligosaccharyltransferase 78:112 Molecular basis of lipid-linked oligosaccharide recognition and processing by bacterial oligosaccharyltransferase. 29058712 4 55 theme N-terminal 634:643 arg1 half 645:648 the N-terminal half 630:648 the N-terminal half of EL5 630:655 Whereas the N-terminal half of EL5 binds LLO, the C-terminal half interacts with the acceptor peptide. 29058712 1 56 theme side 274:277 arg1 chains 279:284 asparagine side chains 263:284 asparagine side chains 263:284 Oligosaccharyltransferase (OST) is a membrane-integral enzyme that catalyzes the transfer of glycans from lipid-linked oligosaccharides (LLOs) onto asparagine side chains, the first step in protein N-glycosylation. 29058712 2 57 theme LLO 499:501 arg1 analog 503:508 a synthetic LLO analog 487:508 a synthetic LLO analog 487:508 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 29058712 2 58 theme OST 386:388 arg1 PglB 391:394 a single-subunit OST, PglB 369:394 PglB 391:394 Here, we report the X-ray structure of a single-subunit OST, PglB from Campylobacter lari, trapped in an intermediate state bound to an acceptor peptide and a synthetic LLO analog. 28271226 2 0 theme results 584:590 arg1 quantitation 564:575 quantitation 564:575 quantitation of the results obtained 564:599 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 4 1 theme acids 992:996 arg1 composition 952:962 different overall composition 934:962 different overall composition 934:962 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 4 1 theme acids 992:996 arg1 number 975:980 different number 965:980 different number of sialic acids 965:996 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 4 1 theme acids 992:996 arg1 size 1022:1025 strongly differing size 1003:1025 strongly differing size without compensation for their very different molar responses 1003:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 2 2 theme carbon 324:329 arg1 chromatography 331:344 Porous graphitic carbon chromatography 307:344 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection 307:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 7 3 theme relative 1449:1456 arg1 abundance 1462:1470 desirable (grey bars) and real (green bars) relative ion abundance 1405:1470 desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS 1405:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 1 4 from structures 281:290 arg1 sample 299:304 one sample 295:304 one sample 295:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 1 5 theme absolute 248:255 arg1 structures 281:290 the various structures 269:290 the various structures in one sample 269:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 1 5 theme absolute 248:255 arg1 amounts 258:264 the relative, if not absolute, amounts 227:264 the relative, if not absolute, amounts of the various structures in one sample 227:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 7 6 theme grey 1416:1419 arg1 bars 1421:1424 grey bars 1416:1424 grey bars 1416:1424 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 4 7 theme molar 1073:1077 arg1 responses 1079:1087 their very different molar responses 1052:1087 their very different molar responses 1052:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 7 8 from abundance 1462:1470 arg1 ESI-MS 1521:1526 positive mode ESI-MS 1507:1526 positive mode ESI-MS 1507:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 3 9 theme type 671:674 arg1 N-glycans 676:684 complex type N-glycans 663:684 complex type N-glycans 663:684 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 4 10 theme liquid 788:793 arg1 chromatography-ESI-MS 795:815 liquid chromatography-ESI-MS 788:815 liquid chromatography-ESI-MS in positive and negative modes 788:846 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 7 11 theme desirable 1405:1413 arg1 abundance 1462:1470 desirable (grey bars) and real (green bars) relative ion abundance 1405:1470 desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS 1405:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 3 12 theme sialic 704:709 arg1 acids 711:715 zero to four sialic acids 691:715 zero to four sialic acids 691:715 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 1 13 from amounts 258:264 arg1 sample 299:304 one sample 295:304 one sample 295:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 4 14 theme tune 859:862 arg1 mix 864:866 the glyco tune mix 849:866 the glyco tune mix 849:866 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 7 15 theme Graphical 1366:1374 arg1 Discrepancy 1385:1395 Graphical Abstract Discrepancy 1366:1395 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 1366:1527 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 1 16 theme true 211:214 arg1 picture 216:222 a true picture 209:222 a true picture of the relative, if not absolute, amounts of the various structures in one sample 209:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 6 17 theme standard 1263:1270 arg1 Addition 1219:1226 Addition 1219:1226 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose 1219:1309 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 2 18 theme resolving 482:490 arg1 oligosaccharides 501:516 resolving isomeric oligosaccharides 482:516 resolving isomeric oligosaccharides 482:516 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 1 19 theme N-glycans 144:152 arg1 analysis 132:139 the analysis 128:139 the analysis of N-glycans 128:152 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 0 20 theme ionization 78:87 arg1 spectrometry 94:105 electrospray ionization mass spectrometry 65:105 electrospray ionization mass spectrometry 65:105 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 0 21 from Determination 0:12 arg1 spectrometry 94:105 electrospray ionization mass spectrometry 65:105 electrospray ionization mass spectrometry 65:105 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 3 22 theme tune 759:762 arg1 " 767:767 an equimolar "glyco tune mix" 739:767 an equimolar "glyco tune mix" 739:767 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 2 23 theme oligosaccharides 501:516 arg1 potential 469:477 a particularly high potential 449:477 a particularly high potential of resolving isomeric oligosaccharides 449:516 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 2 24 theme spectrometry 388:399 arg1 detection 410:418 electrospray ionization mass spectrometry (ESI-MS) detection 359:418 electrospray ionization mass spectrometry (ESI-MS) detection 359:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 4 25 theme number 975:980 arg1 N-glycans 921:929 N-glycans 921:929 N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses 921:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 2 26 theme ionization 372:381 arg1 ESI-MS 402:407 ESI-MS 402:407 ESI-MS 402:407 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 2 26 theme ionization 372:381 arg1 spectrometry 388:399 electrospray ionization mass spectrometry 359:399 electrospray ionization mass spectrometry (ESI-MS) detection 359:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 5 27 theme N-glycans 1128:1136 arg1 quantitation 1099:1110 Relative quantitation 1090:1110 Relative quantitation of human plasma N-glycans 1090:1136 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 4 28 theme sialic 985:990 arg1 acids 992:996 sialic acids 985:996 sialic acids 985:996 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 0 29 theme true 17:20 arg1 ratios 22:27 true ratios 17:27 true ratios of different N-glycan structures 17:60 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 7 30 theme positive 1507:1514 arg1 ESI-MS 1521:1526 positive mode ESI-MS 1507:1526 positive mode ESI-MS 1507:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 1 31 theme ideal 111:115 arg1 method 117:122 An ideal method 108:122 An ideal method for the analysis of N-glycans 108:152 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 4 32 theme overall 944:950 arg1 composition 952:962 different overall composition 934:962 different overall composition 934:962 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 1 33 theme various 273:279 arg1 structures 281:290 the various structures 269:290 the various structures in one sample 269:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 7 34 theme glycans 1496:1502 arg1 amounts 1485:1491 equimolar amounts 1475:1491 equimolar amounts of glycans 1475:1502 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 7 34 theme glycans 1496:1502 arg1 glycans 1496:1502 glycans 1496:1502 glycans 1496:1502 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 2 35 theme graphitic 314:322 arg1 chromatography 331:344 Porous graphitic carbon chromatography 307:344 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection 307:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 0 36 theme N-glycan 42:49 arg1 structures 51:60 different N-glycan structures 32:60 different N-glycan structures 32:60 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 5 37 theme human 1115:1119 arg1 N-glycans 1128:1136 human plasma N-glycans 1115:1136 human plasma N-glycans 1115:1136 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 7 38 theme amounts 1485:1491 arg1 abundance 1462:1470 desirable (grey bars) and real (green bars) relative ion abundance 1405:1470 desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS 1405:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 4 39 theme size 1022:1025 arg1 N-glycans 921:929 N-glycans 921:929 N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses 921:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 3 40 theme structures 639:648 arg1 range 630:634 a range 628:634 a range of structures 628:648 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 5 41 theme glyco 1203:1207 arg1 mix 1214:1216 this external glyco tune mix 1189:1216 this external glyco tune mix 1189:1216 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 1 42 from sample 299:304 arg1 structures 281:290 the various structures 269:290 the various structures in one sample 269:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 1 42 from sample 299:304 arg1 amounts 258:264 the relative, if not absolute, amounts 227:264 the relative, if not absolute, amounts of the various structures in one sample 227:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 7 43 theme ion 1458:1460 arg1 abundance 1462:1470 desirable (grey bars) and real (green bars) relative ion abundance 1405:1470 desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS 1405:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 3 44 theme complex 663:669 arg1 N-glycans 676:684 complex type N-glycans 663:684 complex type N-glycans 663:684 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 3 45 with Man5 655:658 arg1 acids 711:715 zero to four sialic acids 691:715 zero to four sialic acids 691:715 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 5 46 theme correction 1157:1166 arg1 factors 1168:1174 correction factors 1157:1174 correction factors deduced from this external glyco tune mix 1157:1216 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 6 47 with Addition 1219:1226 arg1 13C-galactose 1297:1309 enzymatically added 13C-galactose 1277:1309 enzymatically added 13C-galactose 1277:1309 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 4 48 theme positive 820:827 arg1 modes 842:846 positive and negative modes 820:846 positive and negative modes 820:846 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 7 49 theme real 1431:1434 arg1 abundance 1462:1470 desirable (grey bars) and real (green bars) relative ion abundance 1405:1470 desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS 1405:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 4 50 theme N-glycans 921:929 arg1 quantitation 905:916 quantitation 905:916 quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses 905:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 1 51 theme isomeric 178:185 arg1 structure 187:195 the isomeric structure 174:195 the isomeric structure 174:195 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 6 52 theme absolute 1318:1325 arg1 quantification 1327:1340 absolute quantification 1318:1340 absolute quantification in the same experiment 1318:1363 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 4 53 theme glyco 853:857 arg1 mix 864:866 the glyco tune mix 849:866 the glyco tune mix 849:866 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 4 54 theme different 1063:1071 arg1 responses 1079:1087 their very different molar responses 1052:1087 their very different molar responses 1052:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 2 55 theme little 523:528 arg1 attention 530:538 little attention 523:538 little attention 523:538 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 7 56 theme Abstract 1376:1383 arg1 Discrepancy 1385:1395 Graphical Abstract Discrepancy 1366:1395 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 1366:1527 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 6 57 theme added 1291:1295 arg1 13C-galactose 1297:1309 enzymatically added 13C-galactose 1277:1309 enzymatically added 13C-galactose 1277:1309 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 4 58 from chromatography-ESI-MS 795:815 arg1 modes 842:846 positive and negative modes 820:846 positive and negative modes 820:846 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 4 59 theme negative 833:840 arg1 modes 842:846 positive and negative modes 820:846 positive and negative modes 820:846 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 1 60 dep not 244:246 arg1 if 241:242 if 241:242 if 241:242 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 3 61 theme glyco 753:757 arg1 " 767:767 an equimolar "glyco tune mix" 739:767 an equimolar "glyco tune mix" 739:767 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 0 62 theme electrospray 65:76 arg1 spectrometry 94:105 electrospray ionization mass spectrometry 65:105 electrospray ionization mass spectrometry 65:105 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 2 63 with method 437:442 arg1 potential 469:477 a particularly high potential 449:477 a particularly high potential of resolving isomeric oligosaccharides 449:516 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 4 64 theme quantitation 905:916 arg1 futility 893:900 the futility 889:900 the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses 889:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 3 65 theme equimolar 742:750 arg1 " 767:767 an equimolar "glyco tune mix" 739:767 an equimolar "glyco tune mix" 739:767 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 0 66 theme mass 89:92 arg1 spectrometry 94:105 electrospray ionization mass spectrometry 65:105 electrospray ionization mass spectrometry 65:105 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 3 67 dep four 699:702 arg1 to 696:697 to 696:697 to 696:697 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 7 68 dep desirable 1405:1413 arg1 bars 1421:1424 grey bars 1416:1424 grey bars 1416:1424 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 7 68 dep desirable 1405:1413 arg1 bars 1443:1446 green bars 1437:1446 green bars 1437:1446 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 2 69 theme high 464:467 arg1 potential 469:477 a particularly high potential 449:477 a particularly high potential of resolving isomeric oligosaccharides 449:516 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 6 70 theme same 1349:1352 arg1 experiment 1354:1363 the same experiment 1345:1363 the same experiment 1345:1363 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 5 71 theme Relative 1090:1097 arg1 quantitation 1099:1110 Relative quantitation 1090:1110 Relative quantitation of human plasma N-glycans 1090:1136 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 3 72 theme mix 764:766 arg1 " 767:767 an equimolar "glyco tune mix" 739:767 an equimolar "glyco tune mix" 739:767 In this work, we isolated a range of structures from Man5 to complex type N-glycans with zero to four sialic acids and blended them into an equimolar "glyco tune mix". 28271226 6 73 from quantification 1327:1340 arg1 experiment 1354:1363 the same experiment 1345:1363 the same experiment 1345:1363 Addition of just one isotope-coded internal standard with enzymatically added 13C-galactose led to absolute quantification in the same experiment. 28271226 2 74 theme isomeric 492:499 arg1 oligosaccharides 501:516 resolving isomeric oligosaccharides 482:516 resolving isomeric oligosaccharides 482:516 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 2 75 theme mass 383:386 arg1 ESI-MS 402:407 ESI-MS 402:407 ESI-MS 402:407 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 2 75 theme mass 383:386 arg1 spectrometry 388:399 electrospray ionization mass spectrometry 359:399 electrospray ionization mass spectrometry (ESI-MS) detection 359:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 4 76 theme different 965:973 arg1 number 975:980 different number 965:980 different number of sialic acids 965:996 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 2 77 theme electrospray 359:370 arg1 ESI-MS 402:407 ESI-MS 402:407 ESI-MS 402:407 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 2 77 theme electrospray 359:370 arg1 spectrometry 388:399 electrospray ionization mass spectrometry 359:399 electrospray ionization mass spectrometry (ESI-MS) detection 359:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 0 78 theme ratios 22:27 arg1 Determination 0:12 Determination 0:12 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 0:106 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 1 79 theme amounts 258:264 arg1 picture 216:222 a true picture 209:222 a true picture of the relative, if not absolute, amounts of the various structures in one sample 209:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 0 80 theme different 32:40 arg1 structures 51:60 different N-glycan structures 32:60 different N-glycan structures 32:60 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 7 81 theme mode 1516:1519 arg1 ESI-MS 1521:1526 positive mode ESI-MS 1507:1526 positive mode ESI-MS 1507:1526 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 4 82 theme different 934:942 arg1 composition 952:962 different overall composition 934:962 different overall composition 934:962 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 0 83 theme structures 51:60 arg1 ratios 22:27 true ratios 17:27 true ratios of different N-glycan structures 17:60 Determination of true ratios of different N-glycan structures in electrospray ionization mass spectrometry. 28271226 1 84 theme structures 281:290 arg1 structures 281:290 the various structures 269:290 the various structures in one sample 269:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 1 84 theme structures 281:290 arg1 amounts 258:264 the relative, if not absolute, amounts 227:264 the relative, if not absolute, amounts of the various structures in one sample 227:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 4 85 theme composition 952:962 arg1 N-glycans 921:929 N-glycans 921:929 N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses 921:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 2 86 theme Porous 307:312 arg1 chromatography 331:344 Porous graphitic carbon chromatography 307:344 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection 307:418 Porous graphitic carbon chromatography coupled with electrospray ionization mass spectrometry (ESI-MS) detection has emerged as a method with a particularly high potential of resolving isomeric oligosaccharides, but little attention has so far been paid to quantitation of the results obtained. 28271226 5 87 theme plasma 1121:1126 arg1 N-glycans 1128:1136 human plasma N-glycans 1115:1136 human plasma N-glycans 1115:1136 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 4 88 theme differing 1012:1020 arg1 size 1022:1025 strongly differing size 1003:1025 strongly differing size without compensation for their very different molar responses 1003:1087 When subjected to liquid chromatography-ESI-MS in positive and negative modes, the glyco tune mix clearly demonstrated the futility of quantitation of N-glycans of different overall composition, different number of sialic acids, and strongly differing size without compensation for their very different molar responses. 28271226 5 89 theme external 1194:1201 arg1 mix 1214:1216 this external glyco tune mix 1189:1216 this external glyco tune mix 1189:1216 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 28271226 7 90 theme equimolar 1475:1483 arg1 amounts 1485:1491 equimolar amounts 1475:1491 equimolar amounts of glycans 1475:1502 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 7 90 theme equimolar 1475:1483 arg1 glycans 1496:1502 glycans 1496:1502 glycans 1496:1502 Graphical Abstract Discrepancy between desirable (grey bars) and real (green bars) relative ion abundance of equimolar amounts of glycans in positive mode ESI-MS. 28271226 1 91 theme relative 231:238 arg1 structures 281:290 the various structures 269:290 the various structures in one sample 269:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 1 91 theme relative 231:238 arg1 amounts 258:264 the relative, if not absolute, amounts 227:264 the relative, if not absolute, amounts of the various structures in one sample 227:304 An ideal method for the analysis of N-glycans would both identify the isomeric structure and deliver a true picture of the relative, if not absolute, amounts of the various structures in one sample. 28271226 5 92 theme tune 1209:1212 arg1 mix 1214:1216 this external glyco tune mix 1189:1216 this external glyco tune mix 1189:1216 Relative quantitation of human plasma N-glycans was performed with correction factors deduced from this external glyco tune mix. 24424471 7 0 theme α-1-6 1313:1317 arg1 fucose 1324:1329 decreased α-1-6 core fucose 1303:1329 decreased α-1-6 core fucose 1303:1329 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 6 1 from upregulation 1098:1109 arg1 cells 1161:1165 the iC3-1 cells 1151:1165 the iC3-1 cells 1151:1165 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 1 2 from effects 185:191 arg1 carcinogenesis 221:234 carcinogenesis 221:234 carcinogenesis 221:234 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 2 from effects 185:191 arg1 functions 207:215 biological functions 196:215 biological functions 196:215 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 7 3 theme high 1332:1335 arg1 mannose 1337:1343 high mannose 1332:1343 high mannose 1332:1343 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 4 theme core 1319:1322 arg1 fucose 1324:1329 decreased α-1-6 core fucose 1303:1329 decreased α-1-6 core fucose 1303:1329 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 1 5 attach attachment 147:156 arg2 glycans 161:167 glycans 161:167 glycans 161:167 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 5 attach attachment 147:156 arg1 proteins 172:179 proteins 172:179 proteins 172:179 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 7 6 theme lectin 1217:1222 arg1 array 1224:1228 The lectin array 1213:1228 The lectin array 1213:1228 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 3 7 from choriocarcinoma 537:551 arg1 alterations 491:501 alterations 491:501 alterations of the glycan structure itself in choriocarcinoma 491:551 However, alterations of the glycan structure itself in choriocarcinoma have not been characterized. 24424471 8 8 theme useful 1532:1537 arg1 information 1539:1549 useful information 1532:1549 useful information for understanding of the disease 1532:1582 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 6 9 theme several 1114:1120 arg1 glycogenes 1137:1146 several characteristic glycogenes 1114:1146 several characteristic glycogenes 1114:1146 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 7 10 from GALNAc 1361:1366 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 11 from mannose 1337:1343 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 2 12 dep diagnosis 386:394 arg1 the 382:384 the 382:384 the 382:384 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 8 13 theme first 1460:1464 arg1 This 1448:1451 This 1448:1451 This 1448:1451 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 8 13 theme first 1460:1464 arg1 report 1466:1471 the first report 1456:1471 the first report of a lectin array analysis in choriocarcinoma 1456:1517 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 4 14 theme choriocarcinoma 638:652 arg1 line 624:627 a new choriocarcinoma cell line 597:627 a new choriocarcinoma cell line 597:627 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 14 theme choriocarcinoma 638:652 arg1 iC3-1 662:666 iC3-1 662:666 iC3-1 662:666 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 14 theme choriocarcinoma 638:652 arg1 cell-1 654:659 induced choriocarcinoma cell-1 630:659 induced choriocarcinoma cell-1 (iC3-1) 630:667 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 7 15 theme decreased 1303:1311 arg1 fucose 1324:1329 decreased α-1-6 core fucose 1303:1329 decreased α-1-6 core fucose 1303:1329 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 16 from acid 1260:1263 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 5 17 theme introduced 1027:1036 arg1 microarray 1045:1054 the recently introduced lectin microarray 1014:1054 the recently introduced lectin microarray 1014:1054 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 4 18 theme induced 630:636 arg1 line 624:627 a new choriocarcinoma cell line 597:627 a new choriocarcinoma cell line 597:627 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 18 theme induced 630:636 arg1 iC3-1 662:666 iC3-1 662:666 iC3-1 662:666 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 18 theme induced 630:636 arg1 cell-1 654:659 induced choriocarcinoma cell-1 630:659 induced choriocarcinoma cell-1 (iC3-1) 630:667 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 5 19 theme glycan 922:927 arg1 profiling 929:937 comprehensive glycan profiling 908:937 comprehensive glycan profiling 908:937 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 5 20 theme lectin 1038:1043 arg1 microarray 1045:1054 the recently introduced lectin microarray 1014:1054 the recently introduced lectin microarray 1014:1054 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 5 21 theme profiling 929:937 arg1 performance 893:903 performance 893:903 performance of comprehensive glycan profiling 893:937 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 4 22 theme pathohistology 696:709 arg1 in vivo 711:717 the clinical pathohistology in vivo 683:717 the clinical pathohistology in vivo 683:717 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 7 23 from Galβ1-4GlcNAc 1266:1278 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 24 theme increased 1237:1245 arg1 acid 1260:1263 increased α-2-6-sialic acid 1237:1263 increased α-2-6-sialic acid 1237:1263 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 0 25 theme Glycan 0:5 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling of gestational choriocarcinoma 0:46 Glycan profiling of gestational choriocarcinoma using a lectin microarray. 24424471 2 26 theme disease 427:433 arg1 diagnosis 386:394 diagnosis 386:394 diagnosis 386:394 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 2 26 theme disease 427:433 arg1 treatment 400:408 treatment 400:408 treatment 400:408 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 8 27 theme analysis 1491:1498 arg1 This 1448:1451 This 1448:1451 This 1448:1451 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 8 27 theme analysis 1491:1498 arg1 report 1466:1471 the first report 1456:1471 the first report of a lectin array analysis in choriocarcinoma 1456:1517 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 3 28 from structure 517:525 arg1 choriocarcinoma 537:551 choriocarcinoma 537:551 choriocarcinoma 537:551 However, alterations of the glycan structure itself in choriocarcinoma have not been characterized. 24424471 5 29 theme conventional 986:997 arg1 microarray 999:1008 a conventional microarray 984:1008 a conventional microarray 984:1008 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 2 30 theme trophoblastic 413:425 arg1 disease 427:433 trophoblastic disease 413:433 trophoblastic disease 413:433 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 2 31 gly glycoprotein 281:292 arg1 glycoprotein 281:292 a glycoprotein hormone 279:300 a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors 279:360 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 4 32 theme new 599:601 arg1 line 624:627 a new choriocarcinoma cell line 597:627 a new choriocarcinoma cell line 597:627 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 32 theme new 599:601 arg1 cell-1 654:659 induced choriocarcinoma cell-1 630:659 induced choriocarcinoma cell-1 (iC3-1) 630:667 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 5 33 theme comprehensive 908:920 arg1 profiling 929:937 comprehensive glycan profiling 908:937 comprehensive glycan profiling 908:937 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 1 34 theme important 95:103 arg1 Glycosylation 75:87 Glycosylation 75:87 Glycosylation 75:87 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 34 theme important 95:103 arg1 modification 124:135 an important post-translational modification 92:135 an important post-translational modification 92:135 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 35 theme biological 196:205 arg1 functions 207:215 biological functions 196:215 biological functions 196:215 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 0 36 theme gestational 20:30 arg1 choriocarcinoma 32:46 gestational choriocarcinoma 20:46 gestational choriocarcinoma 20:46 Glycan profiling of gestational choriocarcinoma using a lectin microarray. 24424471 7 37 from GalNacβ1-4Gal 1346:1358 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 1 38 theme post-translational 105:122 arg1 Glycosylation 75:87 Glycosylation 75:87 Glycosylation 75:87 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 38 theme post-translational 105:122 arg1 modification 124:135 an important post-translational modification 92:135 an important post-translational modification 92:135 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 2 39 theme trophoblastic 341:353 arg1 tumors 355:360 trophoblastic tumors 341:360 trophoblastic tumors 341:360 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 4 40 theme cell 619:622 arg1 line 624:627 a new choriocarcinoma cell line 597:627 a new choriocarcinoma cell line 597:627 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 40 theme cell 619:622 arg1 cell-1 654:659 induced choriocarcinoma cell-1 630:659 induced choriocarcinoma cell-1 (iC3-1) 630:667 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 7 41 from GlcNAcβ1-3GalNAc 1281:1296 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 42 theme choriocarcinoma 1399:1413 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 7 43 theme α-2-6-sialic 1247:1258 arg1 acid 1260:1263 increased α-2-6-sialic acid 1237:1263 increased α-2-6-sialic acid 1237:1263 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 2 44 theme reduced 449:455 arg1 incidence 457:465 reduced incidence 449:465 reduced incidence 449:465 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 5 45 theme glycan 820:825 arg1 structures 827:836 glycan structures 820:836 glycan structures 820:836 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 4 46 theme choriocarcinoma 603:617 arg1 line 624:627 a new choriocarcinoma cell line 597:627 a new choriocarcinoma cell line 597:627 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 46 theme choriocarcinoma 603:617 arg1 cell-1 654:659 induced choriocarcinoma cell-1 630:659 induced choriocarcinoma cell-1 (iC3-1) 630:667 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 2 47 theme chorionic 255:263 arg1 hormone 294:300 a glycoprotein hormone 279:300 a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors 279:360 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 2 47 theme chorionic 255:263 arg1 gonadotropin 265:276 human chorionic gonadotropin 249:276 human chorionic gonadotropin 249:276 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 6 48 theme iC3-1 1155:1159 arg1 cells 1161:1165 the iC3-1 cells 1151:1165 the iC3-1 cells 1151:1165 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 6 49 theme glycogenes 1137:1146 arg1 upregulation 1098:1109 significant upregulation 1086:1109 significant upregulation of several characteristic glycogenes in the iC3-1 cells 1086:1165 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 0 50 theme choriocarcinoma 32:46 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling of gestational choriocarcinoma 0:46 Glycan profiling of gestational choriocarcinoma using a lectin microarray. 24424471 5 51 theme structures 827:836 arg1 alterations 805:815 the alterations 801:815 the alterations of glycan structures in the development of choriocarcinoma 801:874 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 2 52 theme human 249:253 arg1 hormone 294:300 a glycoprotein hormone 279:300 a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors 279:360 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 2 52 theme human 249:253 arg1 gonadotropin 265:276 human chorionic gonadotropin 249:276 human chorionic gonadotropin 249:276 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 5 53 theme choriocarcinoma 860:874 arg1 development 845:855 the development 841:855 the development of choriocarcinoma 841:874 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 0 54 theme lectin 56:61 arg1 microarray 63:72 a lectin microarray 54:72 a lectin microarray 54:72 Glycan profiling of gestational choriocarcinoma using a lectin microarray. 24424471 6 55 theme characteristic 1122:1135 arg1 glycogenes 1137:1146 several characteristic glycogenes 1114:1146 several characteristic glycogenes 1114:1146 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 1 56 contain has 181:183 arg1 attachment 147:156 attachment 147:156 attachment of glycans to proteins 147:179 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 1 56 contain has 181:183 arg2 effects 185:191 effects 185:191 effects on biological functions and carcinogenesis 185:234 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 8 57 theme array 1485:1489 arg1 analysis 1491:1498 a lectin array analysis 1476:1498 a lectin array analysis 1476:1498 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 3 58 from alterations 491:501 arg1 choriocarcinoma 537:551 choriocarcinoma 537:551 choriocarcinoma 537:551 However, alterations of the glycan structure itself in choriocarcinoma have not been characterized. 24424471 5 59 theme iC3-1 966:970 arg1 cells 972:976 iC3-1 cells 966:976 iC3-1 cells using a conventional microarray and the recently introduced lectin microarray 966:1054 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 5 60 from alterations 805:815 arg1 development 845:855 the development 841:855 the development of choriocarcinoma 841:874 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 8 61 theme lectin 1478:1483 arg1 analysis 1491:1498 a lectin array analysis 1476:1498 a lectin array analysis 1476:1498 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 7 62 from Galβ1-3Gal 1385:1394 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 2 63 theme glycoprotein 281:292 arg1 hormone 294:300 a glycoprotein hormone 279:300 a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors 279:360 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 2 63 theme glycoprotein 281:292 arg1 gonadotropin 265:276 human chorionic gonadotropin 249:276 human chorionic gonadotropin 249:276 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 3 64 theme glycan 510:515 arg1 structure 517:525 the glycan structure 506:525 the glycan structure itself in choriocarcinoma 506:551 However, alterations of the glycan structure itself in choriocarcinoma have not been characterized. 24424471 6 65 theme Microarray 1057:1066 arg1 comparison 1068:1077 Microarray comparison 1057:1077 Microarray comparison 1057:1077 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 7 66 from fucose 1324:1329 arg1 tissue 1415:1420 choriocarcinoma tissue 1399:1420 choriocarcinoma tissue 1399:1420 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 2 67 theme placental 314:322 arg1 trophoblasts 324:335 placental trophoblasts 314:335 placental trophoblasts 314:335 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 3 68 theme structure 517:525 arg1 alterations 491:501 alterations 491:501 alterations of the glycan structure itself in choriocarcinoma 491:551 However, alterations of the glycan structure itself in choriocarcinoma have not been characterized. 24424471 4 69 theme choriocarcinoma 769:783 arg1 pathogenesis 753:764 pathogenesis 753:764 pathogenesis 753:764 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 4 69 theme choriocarcinoma 769:783 arg1 tumorigenesis 735:747 tumorigenesis 735:747 tumorigenesis 735:747 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 6 70 theme parental 1186:1193 arg1 cells 1206:1210 the parental HTR8/SVneo cells 1182:1210 the parental HTR8/SVneo cells 1182:1210 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 7 71 theme normal 1434:1439 arg1 villi 1441:1445 normal villi 1434:1445 normal villi 1434:1445 The lectin array showed increased α-2-6-sialic acid, Galβ1-4GlcNAc, GlcNAcβ1-3GalNAc, and decreased α-1-6 core fucose, high mannose, GalNacβ1-4Gal, GALNAc (Tn antigen) and Galβ1-3Gal in choriocarcinoma tissue compared to normal villi. 24424471 8 72 from report 1466:1471 arg1 choriocarcinoma 1503:1517 choriocarcinoma 1503:1517 choriocarcinoma 1503:1517 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 5 73 theme clinical 942:949 arg1 samples 951:957 clinical samples 942:957 clinical samples 942:957 In this study, the alterations of glycan structures in the development of choriocarcinoma were examined by performance of comprehensive glycan profiling in clinical samples and in iC3-1 cells using a conventional microarray and the recently introduced lectin microarray. 24424471 4 74 theme clinical 687:694 arg1 in vivo 711:717 the clinical pathohistology in vivo 683:717 the clinical pathohistology in vivo 683:717 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 8 75 theme disease 1576:1582 arg1 understanding 1555:1567 understanding 1555:1567 understanding of the disease 1555:1582 This is the first report of a lectin array analysis in choriocarcinoma and provides useful information for understanding of the disease. 24424471 6 76 theme HTR8/SVneo 1195:1204 arg1 cells 1206:1210 the parental HTR8/SVneo cells 1182:1210 the parental HTR8/SVneo cells 1182:1210 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24424471 2 77 theme gonadotropin 265:276 arg1 Analysis 237:244 Analysis 237:244 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, 237:361 Analysis of human chorionic gonadotropin, a glycoprotein hormone produced by placental trophoblasts and trophoblastic tumors, has contributed to the diagnosis and treatment of trophoblastic disease, resulting in reduced incidence and mortality. 24424471 1 78 theme glycans 161:167 arg1 attachment 147:156 attachment 147:156 attachment of glycans to proteins 147:179 Glycosylation is an important post-translational modification, in which attachment of glycans to proteins has effects on biological functions and carcinogenesis. 24424471 4 79 dep tumorigenesis 735:747 arg1 the 731:733 the 731:733 the 731:733 We established a new choriocarcinoma cell line, induced choriocarcinoma cell-1 (iC3-1), which mimics the clinical pathohistology in vivo, to examine the tumorigenesis and pathogenesis of choriocarcinoma. 24424471 6 80 theme significant 1086:1096 arg1 upregulation 1098:1109 significant upregulation 1086:1109 significant upregulation of several characteristic glycogenes in the iC3-1 cells 1086:1165 Microarray comparison showed significant upregulation of several characteristic glycogenes in the iC3-1 cells as compared to the parental HTR8/SVneo cells. 24978019 6 0 theme computational 883:895 arg1 framework 897:905 this computational framework 878:905 this computational framework 878:905 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 3 1 theme cell/tissue 531:541 arg1 systems 543:549 specific cell/tissue systems 522:549 specific cell/tissue systems 522:549 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 12 2 theme implemented 2096:2106 arg1 features 2108:2115 All the implemented features 2088:2115 All the implemented features 2088:2115 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 2 theme implemented 2096:2106 arg1 part 2133:2136 part 2133:2136 part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2133:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 3 3 theme systems 543:549 arg1 glycome 511:517 glycome 511:517 glycome 511:517 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 3 3 theme systems 543:549 arg1 distribution 495:506 the glycan distribution 484:506 the glycan distribution 484:506 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 10 4 link N-linked 1797:1804 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 5 theme N-linked 1797:1804 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 6 6 theme detailed 961:968 arg1 linkage 1023:1029 linkage 1023:1029 linkage 1023:1029 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 6 theme detailed 961:968 arg1 specificity 1045:1055 substrate specificity 1035:1055 substrate specificity 1035:1055 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 6 theme detailed 961:968 arg1 group 1016:1020 enzymatic functional group 995:1020 enzymatic functional group 995:1020 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 6 theme detailed 961:968 arg1 data 982:985 detailed specificity data 961:985 detailed specificity data such as enzymatic functional group, linkage and substrate specificity 961:1055 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 11 7 theme network 1978:1984 arg1 construction 1992:2003 automated glycosylation network model construction 1954:2003 automated glycosylation network model construction 1954:2003 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 8 8 theme subset 1420:1425 arg1 models 1427:1432 subset models 1420:1432 subset models to identify rate-limiting steps regulating glycan biosynthesis 1420:1495 In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. 24978019 10 9 theme automated 1787:1795 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 8 10 theme graph 1292:1296 arg1 theory 1298:1303 graph theory 1292:1303 graph theory 1292:1303 In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. 24978019 10 11 theme O-linked 1698:1705 arg1 biosynthesis 1714:1725 O-linked glycan biosynthesis 1698:1725 O-linked glycan biosynthesis during the construction of functional selectin-ligands 1698:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 12 dep based 1890:1894 arg1 glycomics 1880:1888 glycomics 1880:1888 glycomics based MS data 1880:1902 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 12 dep based 1890:1894 arg1 data 1899:1902 data 1899:1902 data 1899:1902 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 12 dep based 1890:1894 arg1 MS 1896:1897 MS 1896:1897 MS 1896:1897 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 7 13 theme full 1157:1160 arg1 reconstruction 1170:1183 automated full network reconstruction 1147:1183 automated full network reconstruction 1147:1183 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 10 14 theme glycomics 1880:1888 arg1 biosynthesis 1714:1725 O-linked glycan biosynthesis 1698:1725 O-linked glycan biosynthesis during the construction of functional selectin-ligands 1698:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 14 theme glycomics 1880:1888 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 14 theme glycomics 1880:1888 arg1 handling 1855:1862 handling 1855:1862 handling 1855:1862 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 14 theme glycomics 1880:1888 arg1 analysis 1868:1875 analysis 1868:1875 analysis 1868:1875 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 9 15 theme reaction 1558:1565 arg1 networks 1567:1574 biochemical reaction networks 1546:1574 biochemical reaction networks using mass spectrometry (MS) data 1546:1608 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 11 16 theme glycan 2046:2051 arg1 structure 2053:2061 glycan structure 2046:2061 glycan structure 2046:2061 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 1 17 theme common 125:130 arg1 modifications 163:175 the most common and complex post-translational modifications 116:175 the most common and complex post-translational modifications identified to date 116:194 Glycosylation is among the most common and complex post-translational modifications identified to date. 24978019 6 18 theme functional 1005:1014 arg1 group 1016:1020 enzymatic functional group 995:1020 enzymatic functional group 995:1020 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 7 19 theme single-reaction 1117:1131 arg1 inference 1133:1141 single-reaction inference 1117:1141 single-reaction inference 1117:1141 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 9 20 theme spectrometry 1587:1598 arg1 data 1605:1608 mass spectrometry (MS) data 1582:1608 mass spectrometry (MS) data 1582:1608 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 12 21 theme Glycosylation 2145:2157 arg1 toolbox 2243:2249 an open-source, platform-independent, MATLAB based toolbox 2192:2249 an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2192:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 21 theme Glycosylation 2145:2157 arg1 GNAT 2185:2188 GNAT 2185:2188 GNAT 2185:2188 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 21 theme Glycosylation 2145:2157 arg1 Toolbox 2176:2182 the Glycosylation Network Analysis Toolbox 2141:2182 the Glycosylation Network Analysis Toolbox (GNAT) 2141:2189 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 0 22 theme glycosylation 60:72 arg1 networks 83:90 glycosylation reaction networks 60:90 glycosylation reaction networks 60:90 A computational framework for the automated construction of glycosylation reaction networks. 24978019 11 23 theme biochemistry 2074:2085 arg1 knowledge 2033:2041 knowledge 2033:2041 knowledge of glycan structure and enzyme biochemistry 2033:2085 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 3 24 theme enzymes 457:463 arg1 specificity 436:446 specificity 436:446 specificity 436:446 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 3 24 theme enzymes 457:463 arg1 level 426:430 expression level 415:430 expression level 415:430 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 12 25 theme Analysis 2167:2174 arg1 toolbox 2243:2249 an open-source, platform-independent, MATLAB based toolbox 2192:2249 an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2192:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 25 theme Analysis 2167:2174 arg1 GNAT 2185:2188 GNAT 2185:2188 GNAT 2185:2188 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 25 theme Analysis 2167:2174 arg1 Toolbox 2176:2182 the Glycosylation Network Analysis Toolbox 2141:2182 the Glycosylation Network Analysis Toolbox (GNAT) 2141:2189 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 0 26 theme networks 83:90 arg1 construction 44:55 the automated construction 30:55 the automated construction of glycosylation reaction networks 30:90 A computational framework for the automated construction of glycosylation reaction networks. 24978019 1 27 theme complex 136:142 arg1 modifications 163:175 the most common and complex post-translational modifications 116:175 the most common and complex post-translational modifications identified to date 116:194 Glycosylation is among the most common and complex post-translational modifications identified to date. 24978019 7 28 from present 1256:1262 arg1 system 1271:1276 the system 1267:1276 the system 1267:1276 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 5 29 theme additional 792:801 arg1 methodologies 803:815 additional methodologies 792:815 additional methodologies 792:815 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 3 30 theme expression 415:424 arg1 level 426:430 expression level 415:430 expression level 415:430 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 10 31 theme glycan 1707:1712 arg1 biosynthesis 1714:1725 O-linked glycan biosynthesis 1698:1725 O-linked glycan biosynthesis during the construction of functional selectin-ligands 1698:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 32 theme functional 1754:1763 arg1 selectin-ligands 1765:1780 functional selectin-ligands 1754:1780 functional selectin-ligands 1754:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 7 33 theme present 1256:1262 arg1 enzymes 1248:1254 the enzymes 1244:1254 the enzymes present in the system 1244:1276 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 10 34 theme pathway 1820:1826 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 35 link O-linked 1698:1705 arg1 biosynthesis 1714:1725 O-linked glycan biosynthesis 1698:1725 O-linked glycan biosynthesis during the construction of functional selectin-ligands 1698:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 5 36 theme glycosylation 842:854 arg1 networks 865:872 glycosylation reaction networks 842:872 glycosylation reaction networks 842:872 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 12 37 dep open-source 2195:2205 arg1 platform-independent 2208:2227 platform-independent 2208:2227 platform-independent 2208:2227 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 37 dep open-source 2195:2205 arg1 based 2237:2241 based 2237:2241 based 2237:2241 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 8 38 theme glycan 1477:1482 arg1 biosynthesis 1484:1495 glycan biosynthesis 1477:1495 glycan biosynthesis 1477:1495 In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. 24978019 11 39 theme new 1918:1920 arg1 framework 1936:1944 the new computational framework 1914:1944 the new computational framework 1914:1944 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 6 40 theme enzyme 912:917 arg1 class 919:923 the enzyme class 908:923 the enzyme class 908:923 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 2 41 theme catalytic 221:229 arg1 action 231:236 the catalytic action 217:236 the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans 217:408 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 12 42 theme Systems 2266:2272 arg1 studies 2255:2261 studies 2255:2261 studies of Systems Glycobiology 2255:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 4 43 theme cellular 621:628 arg1 networks 639:646 cellular reaction networks 621:646 cellular reaction networks 621:646 Currently, there is no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation. 24978019 11 44 theme automated 1954:1962 arg1 construction 1992:2003 automated glycosylation network model construction 1954:2003 automated glycosylation network model construction 1954:2003 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 5 45 theme reaction 856:863 arg1 networks 865:872 glycosylation reaction networks 842:872 glycosylation reaction networks 842:872 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 6 46 theme enzymatic 995:1003 arg1 group 1016:1020 enzymatic functional group 995:1020 enzymatic functional group 995:1020 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 7 47 theme reactants 1207:1215 arg1 list 1199:1202 a list 1197:1202 a list of reactants and/or products 1197:1231 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 3 48 theme specific 522:529 arg1 systems 543:549 specific cell/tissue systems 522:549 specific cell/tissue systems 522:549 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 6 49 theme specificity 970:980 arg1 linkage 1023:1029 linkage 1023:1029 linkage 1023:1029 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 49 theme specificity 970:980 arg1 specificity 1045:1055 substrate specificity 1035:1055 substrate specificity 1035:1055 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 49 theme specificity 970:980 arg1 group 1016:1020 enzymatic functional group 995:1020 enzymatic functional group 995:1020 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 6 49 theme specificity 970:980 arg1 data 982:985 detailed specificity data 961:985 detailed specificity data such as enzymatic functional group, linkage and substrate specificity 961:1055 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 2 50 theme sugar 379:383 arg1 residues 385:392 sugar residues 379:392 sugar residues 379:392 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 8 51 used used 1308:1311 arg2 theory 1298:1303 graph theory 1292:1303 graph theory 1292:1303 In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. 24978019 10 52 theme case 1668:1671 arg1 studies 1673:1679 three case studies 1662:1679 three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data 1662:1902 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 5 53 theme machine-readable 730:745 arg1 definition 747:756 a streamlined machine-readable definition 716:756 a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies 716:815 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 6 54 theme substrate 1035:1043 arg1 specificity 1045:1055 substrate specificity 1035:1055 substrate specificity 1035:1055 In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. 24978019 7 55 from system 1271:1276 arg1 present 1256:1262 present 1256:1262 present 1256:1262 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 7 56 theme automated 1147:1155 arg1 reconstruction 1170:1183 automated full network reconstruction 1147:1183 automated full network reconstruction 1147:1183 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 4 57 dep enzymes 609:615 arg1 the 605:607 the 605:607 the 605:607 Currently, there is no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation. 24978019 11 58 theme structure 2053:2061 arg1 knowledge 2033:2041 knowledge 2033:2041 knowledge of glycan structure and enzyme biochemistry 2033:2085 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 9 59 theme biochemical 1546:1556 arg1 networks 1567:1574 biochemical reaction networks 1546:1574 biochemical reaction networks using mass spectrometry (MS) data 1546:1608 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 12 60 theme open-source 2195:2205 arg1 toolbox 2243:2249 an open-source, platform-independent, MATLAB based toolbox 2192:2249 an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2192:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 60 theme open-source 2195:2205 arg1 Toolbox 2176:2182 the Glycosylation Network Analysis Toolbox 2141:2182 the Glycosylation Network Analysis Toolbox (GNAT) 2141:2189 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 2 61 theme multiple 241:248 arg1 families 257:264 multiple enzyme families 241:264 multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans 241:408 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 9 62 theme networks 1567:1574 arg1 synthesis 1533:1541 the synthesis 1529:1541 the synthesis of biochemical reaction networks using mass spectrometry (MS) data 1529:1608 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 7 63 theme network 1162:1168 arg1 reconstruction 1170:1183 automated full network reconstruction 1147:1183 automated full network reconstruction 1147:1183 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 12 64 theme Toolbox 2176:2182 arg1 features 2108:2115 All the implemented features 2088:2115 All the implemented features 2088:2115 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 64 theme Toolbox 2176:2182 arg1 part 2133:2136 part 2133:2136 part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2133:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 7 65 theme associated 1084:1093 arg1 functions 1095:1103 their associated functions 1078:1103 their associated functions 1078:1103 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 9 66 theme mass 1582:1585 arg1 MS 1601:1602 MS 1601:1602 MS 1601:1602 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 9 66 theme mass 1582:1585 arg1 spectrometry 1587:1598 mass spectrometry 1582:1598 mass spectrometry (MS) data 1582:1608 Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. 24978019 10 67 dep handling 1855:1862 arg1 the 1851:1853 the 1851:1853 the 1851:1853 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 67 dep handling 1855:1862 arg1 iii 1846:1848 iii 1846:1848 iii 1846:1848 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 0 68 theme reaction 74:81 arg1 networks 83:90 glycosylation reaction networks 60:90 glycosylation reaction networks 60:90 A computational framework for the automated construction of glycosylation reaction networks. 24978019 11 69 theme enzyme 2067:2072 arg1 biochemistry 2074:2085 enzyme biochemistry 2067:2085 enzyme biochemistry 2067:2085 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 1 70 theme post-translational 144:161 arg1 modifications 163:175 the most common and complex post-translational modifications 116:175 the most common and complex post-translational modifications identified to date 116:194 Glycosylation is among the most common and complex post-translational modifications identified to date. 24978019 7 71 theme new 1062:1064 arg1 classes 1066:1072 The new classes 1058:1072 The new classes 1058:1072 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 12 72 theme Network 2159:2165 arg1 toolbox 2243:2249 an open-source, platform-independent, MATLAB based toolbox 2192:2249 an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology 2192:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 72 theme Network 2159:2165 arg1 GNAT 2185:2188 GNAT 2185:2188 GNAT 2185:2188 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 12 72 theme Network 2159:2165 arg1 Toolbox 2176:2182 the Glycosylation Network Analysis Toolbox 2141:2182 the Glycosylation Network Analysis Toolbox (GNAT) 2141:2189 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 5 73 theme glycosylating 766:778 arg1 enzymes 780:786 the glycosylating enzymes 762:786 the glycosylating enzymes 762:786 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 2 74 theme growing 332:338 arg1 glycans 340:346 growing glycans 332:346 growing glycans 332:346 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 3 75 dep level 426:430 arg1 The 411:413 The 411:413 The 411:413 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 10 76 theme selectin-ligands 1765:1780 arg1 construction 1738:1749 the construction 1734:1749 the construction of functional selectin-ligands 1734:1780 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 0 77 theme computational 2:14 arg1 framework 16:24 A computational framework 0:24 A computational framework for the automated construction of glycosylation reaction networks 0:90 A computational framework for the automated construction of glycosylation reaction networks. 24978019 4 78 theme systematic 575:584 arg1 method 586:591 no systematic method 572:591 no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation 572:674 Currently, there is no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation. 24978019 5 79 theme streamlined 718:728 arg1 definition 747:756 a streamlined machine-readable definition 716:756 a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies 716:815 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 7 80 attach present 1256:1262 arg1 system 1271:1276 the system 1267:1276 the system 1267:1276 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 7 80 attach present 1256:1262 arg2 enzymes 1248:1254 the enzymes 1244:1254 the enzymes present in the system 1244:1276 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 5 81 gly glycosylating 766:778 arg0 enzymes 780:786 the glycosylating enzymes 762:786 the glycosylating enzymes 762:786 To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. 24978019 11 82 theme model 1986:1990 arg1 construction 1992:2003 automated glycosylation network model construction 1954:2003 automated glycosylation network model construction 1954:2003 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 2 83 theme families 257:264 arg1 action 231:236 the catalytic action 217:236 the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans 217:408 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 3 84 theme glycan 488:493 arg1 distribution 495:506 the glycan distribution 484:506 the glycan distribution 484:506 The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. 24978019 0 85 theme automated 34:42 arg1 construction 44:55 the automated construction 30:55 the automated construction of glycosylation reaction networks 30:90 A computational framework for the automated construction of glycosylation reaction networks. 24978019 8 86 theme rate-limiting 1446:1458 arg1 steps 1460:1464 rate-limiting steps 1446:1464 rate-limiting steps regulating glycan biosynthesis 1446:1495 In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. 24978019 7 87 theme products 1224:1231 arg1 list 1199:1202 a list 1197:1202 a list of reactants and/or products 1197:1231 The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. 24978019 11 88 theme computational 1922:1934 arg1 framework 1936:1944 the new computational framework 1914:1944 the new computational framework 1914:1944 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 2 89 theme enzyme 250:255 arg1 families 257:264 multiple enzyme families 241:264 multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans 241:408 It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. 24978019 4 90 theme reaction 630:637 arg1 networks 639:646 cellular reaction networks 621:646 cellular reaction networks 621:646 Currently, there is no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation. 24978019 11 91 theme glycosylation 1964:1976 arg1 construction 1992:2003 automated glycosylation network model construction 1954:2003 automated glycosylation network model construction 1954:2003 Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. 24978019 12 92 dep Systems 2266:2272 arg1 Glycobiology 2274:2285 Glycobiology 2274:2285 Glycobiology 2274:2285 All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology. 24978019 10 93 dep biosynthesis 1714:1725 arg1 i 1695:1695 i 1695:1695 i 1695:1695 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 24978019 10 94 theme glycosylation 1806:1818 arg1 construction 1828:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 ii) automated N-linked glycosylation pathway construction 1783:1839 The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. 28177314 6 0 theme two-dimensional 1342:1356 arg1 notation 1365:1372 the familiar two-dimensional glycan notation 1329:1372 the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg) 1329:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 6 1 theme glycan 1358:1363 arg1 notation 1365:1372 the familiar two-dimensional glycan notation 1329:1372 the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg) 1329:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 2 2 theme great 363:367 arg1 importance 369:378 great importance 363:378 great importance for the understanding of these biological processes 363:430 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 4 3 theme unobscured 622:631 arg1 view 643:646 an unobscured molecular view 619:646 an unobscured molecular view of these multipartite scenarios 619:678 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 6 4 theme stacking 1301:1308 arg1 interactions 1310:1321 stacking interactions 1301:1321 stacking interactions 1301:1321 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 2 5 from way 348:350 arg1 able 277:280 able 277:280 able 277:280 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 5 6 theme structural 922:931 arg1 glycoscience 933:944 structural glycoscience 922:944 structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins 922:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 4 7 theme multipartite 657:668 arg1 scenarios 670:678 these multipartite scenarios 651:678 these multipartite scenarios 651:678 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 1 8 gly glycoproteins 202:214 arg1 glycoproteins 202:214 glycoproteins 202:214 glycoproteins 202:214 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 4 9 theme many 795:798 arg1 each 817:820 each 817:820 each 817:820 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 9 theme many 795:798 arg1 sugars 809:814 many branched sugars 795:814 many branched sugars 795:814 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 10 theme potential 889:897 arg1 partners 907:914 more than three potential bonding partners 873:914 more than three potential bonding partners 873:914 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 11 dep need 728:731 arg1 addition 712:719 addition 712:719 addition 712:719 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 3 12 theme additional 577:586 arg1 interactions 597:608 additional stacking interactions 577:608 additional stacking interactions 577:608 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 4 13 theme interacting 745:755 arg1 residues 765:772 the interacting protein residues 741:772 the interacting protein residues 741:772 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 2 14 theme visualise 285:293 arg1 protein-glycan 295:308 visualise protein-glycan 285:308 visualise protein-glycan 285:308 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 4 15 dep each 817:820 arg1 composed 822:829 composed 822:829 composed of more than ten non-H atoms 822:858 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 15 dep each 817:820 arg1 offering 864:871 offering 864:871 offering more than three potential bonding partners 864:914 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 1 16 theme correct 183:189 arg1 folding 191:197 the correct folding 179:197 the correct folding of glycoproteins 179:214 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 4 17 theme molecular 633:641 arg1 view 643:646 an unobscured molecular view 619:646 an unobscured molecular view of these multipartite scenarios 619:678 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 18 theme branched 800:807 arg1 each 817:820 each 817:820 each 817:820 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 18 theme branched 800:807 arg1 sugars 809:814 many branched sugars 795:814 many branched sugars 795:814 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 3 19 theme glycosylation 454:466 arg1 sugars 468:473 glycosylation sugars 454:473 glycosylation sugars 454:473 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 6 20 theme bonding-network 1246:1260 arg1 depiction 1262:1270 a simplified bonding-network depiction 1233:1270 a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) 1233:1322 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 5 21 dep pressing 1132:1139 arg1 ever 1146:1149 ever 1146:1149 ever 1146:1149 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 5 21 dep pressing 1132:1139 arg1 need 1071:1074 the need 1067:1074 the need for a clear way of depicting these interactions 1067:1122 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 5 21 dep pressing 1132:1139 arg1 pressing 1132:1139 pressing 1132:1139 pressing 1132:1139 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 3 22 theme structural 436:445 arg1 terms 447:451 structural terms 436:451 structural terms 436:451 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 1 23 theme close-range 98:108 arg1 essential 165:173 essential 165:173 essential 165:173 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 1 23 theme close-range 98:108 arg1 interactions 110:121 The close-range interactions 94:121 The close-range interactions provided by covalently linked glycans 94:159 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 1 24 theme glycoproteins 202:214 arg1 folding 191:197 the correct folding 179:197 the correct folding of glycoproteins 179:214 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 0 25 theme schematic 15:23 arg1 representation 43:56 a schematic three-dimensional representation 13:56 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 0:92 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 28177314 6 26 theme simplified 1235:1244 arg1 depiction 1262:1270 a simplified bonding-network depiction 1233:1270 a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) 1233:1322 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 3 27 theme stacking 588:595 arg1 interactions 597:608 additional stacking interactions 577:608 additional stacking interactions 577:608 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 5 28 dep glycoscience 933:944 arg1 increasing 986:995 increasing 986:995 steadily increasing the deposition rate of three-dimensional structures of glycoproteins 977:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 5 28 dep glycoscience 933:944 arg1 gaining 954:960 gaining 954:960 finally gaining popularity 946:971 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 2 29 theme glycan-glycan 314:326 arg1 contacts 328:335 glycan-glycan contacts 314:335 glycan-glycan contacts 314:335 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 2 30 from able 277:280 arg1 way 348:350 a clear way 340:350 a clear way 340:350 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 6 31 theme schematic 1159:1167 arg1 representation 1169:1182 a schematic representation 1157:1182 a schematic representation 1157:1182 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 6 32 theme glycobiology 1386:1397 arg1 community 1399:1407 the glycobiology community 1382:1407 the glycobiology community 1382:1407 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 2 33 theme processes 422:430 arg1 understanding 388:400 the understanding 384:400 the understanding of these biological processes 384:430 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 0 34 theme three-dimensional 25:41 arg1 representation 43:56 a schematic three-dimensional representation 13:56 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 0:92 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 28177314 6 35 theme hydrogen 1282:1289 arg1 bonds 1291:1295 hydrogen bonds 1282:1295 hydrogen bonds 1282:1295 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 2 36 theme biological 411:420 arg1 processes 422:430 these biological processes 405:430 these biological processes 405:430 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 1 37 link linked 146:151 arg1 glycans 153:159 covalently linked glycans 135:159 covalently linked glycans 135:159 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 4 38 theme non-H 848:852 arg1 atoms 854:858 more than ten non-H atoms 834:858 more than ten non-H atoms 834:858 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 3 39 from glue 475:478 arg1 terms 447:451 structural terms 436:451 structural terms 436:451 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 3 40 theme hydrogen 505:512 arg1 bonds 514:518 hydrogen bonds 505:518 hydrogen bonds 505:518 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 5 41 gly glycoproteins 1052:1064 arg1 glycoproteins 1052:1064 glycoproteins 1052:1064 glycoproteins 1052:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 4 42 contain contain 787:793 arg1 glycans 775:781 glycans 775:781 glycans 775:781 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 42 contain contain 787:793 arg2 sugars 809:814 many branched sugars 795:814 many branched sugars 795:814 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 4 42 contain contain 787:793 arg2 each 817:820 each 817:820 each 817:820 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 5 43 theme clear 1082:1086 arg1 way 1088:1090 a clear way 1080:1090 a clear way of depicting these interactions 1080:1122 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 4 44 theme bonding 899:905 arg1 partners 907:914 more than three potential bonding partners 873:914 more than three potential bonding partners 873:914 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 6 45 theme familiar 1333:1340 arg1 notation 1365:1372 the familiar two-dimensional glycan notation 1329:1372 the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg) 1329:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 5 46 theme structures 1038:1047 arg1 rate 1012:1015 the deposition rate 997:1015 the deposition rate of three-dimensional structures of glycoproteins 997:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 1 47 theme linked 146:151 arg1 glycans 153:159 covalently linked glycans 135:159 covalently linked glycans 135:159 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 1 48 theme pivotal 232:238 arg1 role 240:243 a pivotal role 230:243 a pivotal role 230:243 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28177314 5 49 theme three-dimensional 1020:1036 arg1 structures 1038:1047 three-dimensional structures 1020:1047 three-dimensional structures of glycoproteins 1020:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 5 50 theme deposition 1001:1010 arg1 rate 1012:1015 the deposition rate 997:1015 the deposition rate of three-dimensional structures of glycoproteins 997:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 2 51 theme clear 342:346 arg1 way 348:350 a clear way 340:350 a clear way 340:350 Being able to visualise protein-glycan and glycan-glycan contacts in a clear way is thus of great importance for the understanding of these biological processes. 28177314 5 52 with pressing 1132:1139 arg1 glycoscience 933:944 structural glycoscience 922:944 structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins 922:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 4 53 theme protein 757:763 arg1 residues 765:772 the interacting protein residues 741:772 the interacting protein residues 741:772 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 6 54 theme molecular 1452:1460 arg1 CCP4mg 1480:1485 CCP4mg 1480:1485 CCP4mg 1480:1485 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 6 54 theme molecular 1452:1460 arg1 project 1471:1477 the CCP4 molecular graphics project 1443:1477 the CCP4 molecular graphics project (CCP4mg) 1443:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 5 55 theme glycoproteins 1052:1064 arg1 structures 1038:1047 three-dimensional structures 1020:1047 three-dimensional structures of glycoproteins 1020:1064 With structural glycoscience finally gaining popularity and steadily increasing the deposition rate of three-dimensional structures of glycoproteins, the need for a clear way of depicting these interactions is more pressing than ever. 28177314 6 56 theme graphics 1462:1469 arg1 CCP4mg 1480:1485 CCP4mg 1480:1485 CCP4mg 1480:1485 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 6 56 theme graphics 1462:1469 arg1 project 1471:1477 the CCP4 molecular graphics project 1443:1477 the CCP4 molecular graphics project (CCP4mg) 1443:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 4 57 theme scenarios 670:678 arg1 view 643:646 an unobscured molecular view 619:646 an unobscured molecular view of these multipartite scenarios 619:678 Finding an unobscured molecular view of these multipartite scenarios is usually far from trivial; in addition to the need to show the interacting protein residues, glycans may contain many branched sugars, each composed of more than ten non-H atoms and offering more than three potential bonding partners. 28177314 6 58 theme CCP4 1447:1450 arg1 CCP4mg 1480:1485 CCP4mg 1480:1485 CCP4mg 1480:1485 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 6 58 theme CCP4 1447:1450 arg1 project 1471:1477 the CCP4 molecular graphics project 1443:1477 the CCP4 molecular graphics project (CCP4mg) 1443:1486 Here a schematic representation, named Glycoblocks, is introduced which combines a simplified bonding-network depiction (covering hydrogen bonds and stacking interactions) with the familiar two-dimensional glycan notation used by the glycobiology community, brought into three dimensions by the CCP4 molecular graphics project (CCP4mg). 28177314 0 59 dep Glycoblocks 0:10 arg1 representation 43:56 a schematic three-dimensional representation 13:56 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 0:92 Glycoblocks: a schematic three-dimensional representation for glycans and their interactions. 28177314 3 60 theme bound 544:548 arg1 glycans 550:556 non-covalently bound glycans 529:556 non-covalently bound glycans 529:556 In structural terms, glycosylation sugars glue the protein together via hydrogen bonds, whereas non-covalently bound glycans frequently harness additional stacking interactions. 28177314 1 61 theme recognition 248:258 arg1 processes 260:268 recognition processes 248:268 recognition processes 248:268 The close-range interactions provided by covalently linked glycans are essential for the correct folding of glycoproteins and also play a pivotal role in recognition processes. 28298443 7 0 theme glycan 1368:1373 arg1 formation 1336:1344 the formation 1332:1344 the formation of cereose-containing glycan 1332:1373 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 7 1 theme Gene 1230:1233 arg1 knockouts 1235:1243 Gene knockouts 1230:1243 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase 1230:1299 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 3 2 with sugars 368:373 arg1 structure 407:415 a 3-C-methyl-6-deoxyhexose structure 380:415 a 3-C-methyl-6-deoxyhexose structure 380:415 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 10 3 theme 3-C-methyl-6-deoxy 1696:1713 arg1 sugars 1715:1720 the uncommon 3-C-methyl-6-deoxy sugars 1683:1720 the uncommon 3-C-methyl-6-deoxy sugars 1683:1720 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 1 4 theme host-pathogen 185:197 arg1 interactions 199:210 host-pathogen interactions 185:210 host-pathogen interactions 185:210 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 2 5 theme bacterial 304:312 arg1 growth 314:319 growth 314:319 growth 314:319 Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. 28298443 3 6 attach linked 422:427 arg2 sugars 368:373 two rare sugars 359:373 two rare sugars with a 3-C-methyl-6-deoxyhexose structure 359:415 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 3 6 attach linked 422:427 arg1 glycans 438:444 spore glycans 432:444 spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876 432:489 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 7 theme mass 646:649 arg1 spectrometry 651:662 mass spectrometry 646:662 mass spectrometry 646:662 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 2 8 theme sugar 241:245 arg1 composition 247:257 the sugar composition 237:257 the sugar composition of bacterial cell surfaces 237:284 Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. 28298443 9 9 theme spore 1587:1591 arg1 resistance 1593:1602 spore resistance 1587:1602 spore resistance to heat 1587:1610 Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. 28298443 4 10 dep activities 618:627 arg1 determined 632:641 determined 632:641 determined by mass spectrometry and one- and two-dimensional NMR methods 632:703 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 10 dep activities 618:627 arg1 CTP 706:708 CTP 706:708 the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 586:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 1 11 from interactions 199:210 arg1 critical 130:137 critical 130:137 critical 130:137 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 2 12 theme cell 272:275 arg1 surfaces 277:284 bacterial cell surfaces 262:284 bacterial cell surfaces 262:284 Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. 28298443 4 13 theme NMR 693:695 arg1 methods 697:703 one- and two-dimensional NMR methods 668:703 one- and two-dimensional NMR methods 668:703 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 7 14 theme cereose-containing 1349:1366 arg1 glycan 1368:1373 cereose-containing glycan 1349:1373 cereose-containing glycan 1349:1373 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 4 15 dep B. 538:539 arg1 cereus 541:546 cereus 541:546 cereus 541:546 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 1 16 theme cell-cell 143:151 arg1 recognition 153:163 recognition 153:163 recognition 153:163 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 0 17 from operon 12:17 arg1 cereus 31:36 Bacillus cereus 22:36 Bacillus cereus 22:36 A four-gene operon in Bacillus cereus produces two rare spore-decorating sugars. 28298443 7 18 dep C-methyltransferase 1261:1279 arg1 the 1248:1250 the 1248:1250 the 1248:1250 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 7 18 dep C-methyltransferase 1261:1279 arg1 Bacillus 1252:1259 the Bacillus C-methyltransferase and the 4-reductase 1248:1299 Bacillus 1252:1259 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 8 19 theme dry 1448:1450 arg1 weight 1452:1457 spore dry weight 1442:1457 0.2-1% spore dry weight 1435:1457 We also found that cereose represented 0.2-1% spore dry weight. 28298443 5 20 theme 4-epimer 983:990 arg1 CDP-cillose 1022:1032 CDP-cillose 1022:1032 CDP-cillose 1022:1032 The last enzyme predominantly yielded CDP-3-C-methyl-6-deoxygulose (CDP-cereose) and likely generated a 4-epimer CDP-3-C-methyl-6-deoxyallose (CDP-cillose). 28298443 5 20 theme 4-epimer 983:990 arg1 CDP-3-C-methyl-6-deoxyallose 992:1019 a 4-epimer CDP-3-C-methyl-6-deoxyallose 981:1019 a 4-epimer CDP-3-C-methyl-6-deoxyallose (CDP-cillose) 981:1033 The last enzyme predominantly yielded CDP-3-C-methyl-6-deoxygulose (CDP-cereose) and likely generated a 4-epimer CDP-3-C-methyl-6-deoxyallose (CDP-cillose). 28298443 6 21 dep produce 1083:1089 arg1 whereas 1180:1186 whereas 1180:1186 whereas 1180:1186 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 6 22 from formation 1129:1137 arg1 spores 1172:1177 spores 1172:1177 spores 1172:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 4 23 theme SAM 792:794 arg1 C-methyltransferase 796:814 NADH-dependent SAM:C-methyltransferase 777:814 NADH-dependent SAM:C-methyltransferase 777:814 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 6 24 theme lato 1072:1075 arg1 group 1077:1081 the B. cereus sensu lato group 1052:1081 the B. cereus sensu lato group 1052:1081 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 4 25 theme CDP-Glc 752:758 arg1 4,6-dehydratase 760:774 CDP-Glc 4,6-dehydratase 752:774 CDP-Glc 4,6-dehydratase 752:774 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 3 26 theme spore 432:436 arg1 glycans 438:444 spore glycans 432:444 spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876 432:489 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 7 27 theme 4-reductase 1289:1299 arg1 knockouts 1235:1243 Gene knockouts 1230:1243 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase 1230:1299 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 1 28 theme glycan 91:96 arg1 structures 98:107 Bacterial glycan structures 81:107 Bacterial glycan structures on cell surfaces 81:124 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 2 29 theme bacterial 262:270 arg1 surfaces 277:284 bacterial cell surfaces 262:284 bacterial cell surfaces 262:284 Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. 28298443 5 30 theme last 883:886 arg1 enzyme 888:893 The last enzyme 879:893 The last enzyme 879:893 The last enzyme predominantly yielded CDP-3-C-methyl-6-deoxygulose (CDP-cereose) and likely generated a 4-epimer CDP-3-C-methyl-6-deoxyallose (CDP-cillose). 28298443 4 31 dep CTP 706:708 arg1 4,6-dehydratase 760:774 CDP-Glc 4,6-dehydratase 752:774 CDP-Glc 4,6-dehydratase 752:774 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 31 dep CTP 706:708 arg1 C-methyltransferase 796:814 NADH-dependent SAM:C-methyltransferase 777:814 NADH-dependent SAM:C-methyltransferase 777:814 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 31 dep CTP 706:708 arg1 4-reductase 866:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 31 dep CTP 706:708 arg1 cytidylyltransferase 730:749 glucose-1-phosphate cytidylyltransferase 710:749 glucose-1-phosphate cytidylyltransferase 710:749 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 6 32 theme glycans 1161:1167 arg1 formation 1129:1137 the formation 1125:1137 the formation of cereose-containing glycans on spores 1125:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 10 33 theme Bacillus 1794:1801 arg1 recognition 1738:1748 cell-surface recognition 1725:1748 cell-surface recognition 1725:1748 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 10 33 theme Bacillus 1794:1801 arg1 interactions 1768:1779 host-pathogen interactions 1754:1779 host-pathogen interactions 1754:1779 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 4 34 theme four-gene 518:526 arg1 operon 528:533 a four-gene operon 516:533 a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 516:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 34 theme four-gene 518:526 arg1 proteins 572:579 proteins 572:579 proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 572:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 0 35 theme rare 51:54 arg1 sugars 73:78 two rare spore-decorating sugars 47:78 two rare spore-decorating sugars 47:78 A four-gene operon in Bacillus cereus produces two rare spore-decorating sugars. 28298443 3 36 dep Bacillus 449:456 arg1 cereus 458:463 cereus 458:463 cereus 458:463 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 37 theme two-dimensional 677:691 arg1 methods 697:703 one- and two-dimensional NMR methods 668:703 one- and two-dimensional NMR methods 668:703 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 38 theme following 590:598 arg1 activities 618:627 the following sequential enzyme activities 586:627 the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 586:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 4 39 theme CDP-3-C-methyl-6-deoxyhexose 837:864 arg1 4-reductase 866:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 3 40 from glycans 438:444 arg1 ATCC 480:483 ATCC 10876 480:489 ATCC 10876 480:489 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 3 40 from glycans 438:444 arg1 14579 470:474 Bacillus cereus ATCC 14579 449:474 Bacillus cereus ATCC 14579 449:474 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 41 theme enzyme 611:616 arg1 activities 618:627 the following sequential enzyme activities 586:627 the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 586:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 6 42 from spores 1172:1177 arg1 formation 1129:1137 the formation 1125:1137 the formation of cereose-containing glycans on spores 1125:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 10 43 theme sugars 1715:1720 arg1 roles 1674:1678 the roles 1670:1678 the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus 1670:1801 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 6 44 theme sensu 1066:1070 arg1 group 1077:1081 the B. cereus sensu lato group 1052:1081 the B. cereus sensu lato group 1052:1081 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 9 45 from resistance 1593:1602 arg1 sporulation 1572:1582 sporulation 1572:1582 sporulation 1572:1582 Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. 28298443 4 46 theme one- 668:671 arg1 methods 697:703 one- and two-dimensional NMR methods 668:703 one- and two-dimensional NMR methods 668:703 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 10 47 theme uncommon 1687:1694 arg1 sugars 1715:1720 the uncommon 3-C-methyl-6-deoxy sugars 1683:1720 the uncommon 3-C-methyl-6-deoxy sugars 1683:1720 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 6 48 from glycans 1161:1167 arg1 spores 1172:1177 spores 1172:1177 spores 1172:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 6 49 theme B. 1056:1057 arg1 group 1077:1081 the B. cereus sensu lato group 1052:1081 the B. cereus sensu lato group 1052:1081 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 7 50 from involvement 1317:1327 arg1 formation 1336:1344 the formation 1332:1344 the formation of cereose-containing glycan 1332:1373 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 7 50 from involvement 1317:1327 arg1 spores 1388:1393 B. cereus spores 1378:1393 B. cereus spores 1378:1393 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 10 51 theme cell-surface 1725:1736 arg1 recognition 1738:1748 cell-surface recognition 1725:1748 cell-surface recognition 1725:1748 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 1 52 theme cell 112:115 arg1 surfaces 117:124 cell surfaces 112:124 cell surfaces 112:124 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 6 53 dep B. 1056:1057 arg1 cereus 1059:1064 cereus 1059:1064 cereus 1059:1064 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 2 54 theme surfaces 277:284 arg1 composition 247:257 the sugar composition 237:257 the sugar composition of bacterial cell surfaces 237:284 Accordingly, unraveling the sugar composition of bacterial cell surfaces can shed light on bacterial growth and pathogenesis. 28298443 4 55 theme glucose-1-phosphate 710:728 arg1 cytidylyltransferase 730:749 glucose-1-phosphate cytidylyltransferase 710:749 glucose-1-phosphate cytidylyltransferase 710:749 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 0 56 theme spore-decorating 56:71 arg1 sugars 73:78 two rare spore-decorating sugars 47:78 two rare spore-decorating sugars 47:78 A four-gene operon in Bacillus cereus produces two rare spore-decorating sugars. 28298443 4 57 with proteins 572:579 arg1 activities 618:627 the following sequential enzyme activities 586:627 the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 586:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 8 58 theme 0.2-1 1435:1439 arg1 % 1440:1440 0.2-1% 1435:1440 0.2-1% spore dry weight 1435:1457 We also found that cereose represented 0.2-1% spore dry weight. 28298443 6 59 theme CDP-3-C-methyl-6-deoxy 1091:1112 arg1 sugars 1114:1119 CDP-3-C-methyl-6-deoxy sugars 1091:1119 CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores 1091:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 8 60 theme spore 1442:1446 arg1 weight 1452:1457 spore dry weight 1442:1457 0.2-1% spore dry weight 1435:1457 We also found that cereose represented 0.2-1% spore dry weight. 28298443 6 61 theme group 1077:1081 arg1 members 1041:1047 Some members 1036:1047 Some members of the B. cereus sensu lato group 1036:1081 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 3 62 theme rare 363:366 arg1 sugars 368:373 two rare sugars 359:373 two rare sugars with a 3-C-methyl-6-deoxyhexose structure 359:415 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 63 theme NADH-dependent 777:790 arg1 C-methyltransferase 796:814 NADH-dependent SAM:C-methyltransferase 777:814 NADH-dependent SAM:C-methyltransferase 777:814 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 3 64 theme Bacillus 449:456 arg1 14579 470:474 Bacillus cereus ATCC 14579 449:474 Bacillus cereus ATCC 14579 449:474 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 0 65 theme four-gene 2:10 arg1 operon 12:17 A four-gene operon 0:17 A four-gene operon in Bacillus cereus 0:36 A four-gene operon in Bacillus cereus produces two rare spore-decorating sugars. 28298443 1 66 from critical 130:137 arg1 interactions 199:210 host-pathogen interactions 185:210 host-pathogen interactions 185:210 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 1 67 theme Bacterial 81:89 arg1 structures 98:107 Bacterial glycan structures 81:107 Bacterial glycan structures on cell surfaces 81:124 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 3 68 theme 3-C-methyl-6-deoxyhexose 382:405 arg1 structure 407:415 a 3-C-methyl-6-deoxyhexose structure 380:415 a 3-C-methyl-6-deoxyhexose structure 380:415 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 69 theme ATCC 548:551 arg1 14579 553:557 B. cereus ATCC 14579 538:557 B. cereus ATCC 14579 538:557 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 10 70 theme host-pathogen 1754:1766 arg1 interactions 1768:1779 host-pathogen interactions 1754:1779 host-pathogen interactions 1754:1779 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 7 71 theme C-methyltransferase 1261:1279 arg1 knockouts 1235:1243 Gene knockouts 1230:1243 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase 1230:1299 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 7 72 dep B. 1378:1379 arg1 cereus 1381:1386 cereus 1381:1386 cereus 1381:1386 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 6 73 theme cereose-containing 1142:1159 arg1 glycans 1161:1167 cereose-containing glycans 1142:1167 cereose-containing glycans on spores 1142:1177 Some members of the B. cereus sensu lato group produce CDP-3-C-methyl-6-deoxy sugars for the formation of cereose-containing glycans on spores, whereas others such as Bacillus anthracis do not. 28298443 3 74 theme ATCC 465:468 arg1 14579 470:474 Bacillus cereus ATCC 14579 449:474 Bacillus cereus ATCC 14579 449:474 Here, we found that two rare sugars with a 3-C-methyl-6-deoxyhexose structure were linked to spore glycans in Bacillus cereus ATCC 14579 and ATCC 10876. 28298443 4 75 from operon 528:533 arg1 14579 553:557 B. cereus ATCC 14579 538:557 B. cereus ATCC 14579 538:557 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 8 76 dep % 1440:1440 arg1 weight 1452:1457 spore dry weight 1442:1457 0.2-1% spore dry weight 1435:1457 We also found that cereose represented 0.2-1% spore dry weight. 28298443 4 77 theme B. 538:539 arg1 14579 553:557 B. cereus ATCC 14579 538:557 B. cereus ATCC 14579 538:557 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 1 78 from structures 98:107 arg1 surfaces 117:124 cell surfaces 112:124 cell surfaces 112:124 Bacterial glycan structures on cell surfaces are critical for cell-cell recognition and adhesion and in host-pathogen interactions. 28298443 4 79 theme sequential 600:609 arg1 activities 618:627 the following sequential enzyme activities 586:627 the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 586:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 10 80 from roles 1674:1678 arg1 recognition 1738:1748 cell-surface recognition 1725:1748 cell-surface recognition 1725:1748 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 10 80 from roles 1674:1678 arg1 interactions 1768:1779 host-pathogen interactions 1754:1779 host-pathogen interactions 1754:1779 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 9 81 from changes 1561:1567 arg1 sporulation 1572:1582 sporulation 1572:1582 sporulation 1572:1582 Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. 28298443 7 82 theme B. 1378:1379 arg1 spores 1388:1393 B. cereus spores 1378:1393 B. cereus spores 1378:1393 Gene knockouts of the Bacillus C-methyltransferase and the 4-reductase confirmed their involvement in the formation of cereose-containing glycan on B. cereus spores. 28298443 10 83 theme new 1652:1654 arg1 insights 1656:1663 new insights 1652:1663 new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus 1652:1801 The findings reported here may provide new insights into the roles of the uncommon 3-C-methyl-6-deoxy sugars in cell-surface recognition and host-pathogen interactions of the genus Bacillus. 28298443 4 84 theme NADPH-dependent 821:835 arg1 4-reductase 866:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase 821:876 Moreover, we identified a four-gene operon in B. cereus ATCC 14579 that encodes proteins with the following sequential enzyme activities as determined by mass spectrometry and one- and two-dimensional NMR methods: CTP:glucose-1-phosphate cytidylyltransferase, CDP-Glc 4,6-dehydratase, NADH-dependent SAM:C-methyltransferase, and NADPH-dependent CDP-3-C-methyl-6-deoxyhexose 4-reductase. 28298443 9 85 theme wild 1521:1524 arg1 type 1526:1529 the wild type 1517:1529 the wild type 1517:1529 Moreover, mutants lacking cereose germinated faster than the wild type, yet the mutants exhibited no changes in sporulation or spore resistance to heat. 25712693 0 0 theme mass 77:80 arg1 spectrometry 82:93 mass spectrometry 77:93 mass spectrometry 77:93 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 3 1 theme N-glycans 473:481 arg1 spectra 462:468 MS/MS spectra 456:468 MS/MS spectra of N-glycans that match diagnostic ions to determine the structures 456:536 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 2 2 from spectra 315:321 arg1 identification 268:281 the identification 264:281 the identification of glycan structures from MS/MS spectra 264:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 2 2 from spectra 315:321 arg1 structures 293:302 glycan structures 286:302 glycan structures from MS/MS spectra 286:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 7 3 theme k-morimt 866:873 arg1 shimadzu.co.jp 875:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 3 4 theme diagnostic 343:352 arg1 ions 354:357 diagnostic ions 343:357 diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) 343:437 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 6 5 theme standard 818:825 arg1 package 827:833 the standard package 814:833 the standard package of Mass++ for Windows 814:855 The GlycanAnalysis plug-in is included in the standard package of Mass++ for Windows. 25712693 7 6 dep shimadzu.co.jp 902:915 arg1 material 983:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 7 7 theme @ 874:874 arg1 shimadzu.co.jp 875:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 7 8 theme INFORMATION 957:967 arg1 material 983:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 7 9 theme nishikaz 893:900 arg1 shimadzu.co.jp 902:915 nishikaz@shimadzu.co.jp 893:915 nishikaz@shimadzu.co.jp 893:915 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 3 10 theme MS/MS 456:460 arg1 spectra 462:468 MS/MS spectra 456:468 MS/MS spectra of N-glycans that match diagnostic ions to determine the structures 456:536 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 5 11 dep AVAILABILITY 648:659 arg1 files 695:699 The executable files 680:699 The executable files 680:699 AVAILABILITY AND IMPLEMENTATION The executable files of Mass++ are available for free at http://www.first-ms3d.jp/english/. 25712693 7 12 theme Supplementary 969:981 arg1 material 983:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 7 13 theme CONTACT 858:864 arg1 shimadzu.co.jp 875:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp 858:888 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 4 14 theme analysis 630:637 arg1 Mass++ 579:584 Mass++ 579:584 Mass++ 579:584 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 4 14 theme analysis 630:637 arg1 program 639:645 analysis program 630:645 analysis program 630:645 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 3 15 theme glycan 396:401 arg1 database 403:410 a glycan database 394:410 a glycan database (GlycomeDB or KEGG GLYCAN) 394:437 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 1 16 dep spectrometry 119:130 arg1 MS 142:143 MS 142:143 MS 142:143 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 16 dep spectrometry 119:130 arg1 MS/MS 133:137 MS/MS 133:137 MS/MS 133:137 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 16 dep spectrometry 119:130 arg1 n 145:145 n 145:145 n 145:145 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 3 17 theme KEGG 426:429 arg1 GLYCAN 431:436 KEGG GLYCAN 426:436 KEGG GLYCAN 426:436 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 3 18 theme structures 369:378 arg1 ions 354:357 diagnostic ions 343:357 diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) 343:437 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 6 19 theme GlycanAnalysis 776:789 arg1 plug-in 791:797 The GlycanAnalysis plug-in 772:797 The GlycanAnalysis plug-in 772:797 The GlycanAnalysis plug-in is included in the standard package of Mass++ for Windows. 25712693 0 20 theme GlycanAnalysis 0:13 arg1 Plug-in 15:21 GlycanAnalysis Plug-in 0:21 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 0:94 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 3 21 dep database 403:410 arg1 GlycomeDB 413:421 GlycomeDB 413:421 GlycomeDB 413:421 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 3 21 dep database 403:410 arg1 GLYCAN 431:436 KEGG GLYCAN 426:436 KEGG GLYCAN 426:436 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 7 22 theme acyshzw 920:926 arg1 shimadzu.co.jp 928:941 acyshzw@shimadzu.co.jp 920:941 acyshzw@shimadzu.co.jp 920:941 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 2 23 theme structures 293:302 arg1 identification 268:281 the identification 264:281 the identification of glycan structures from MS/MS spectra 264:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 7 24 from online 1024:1029 arg1 available 996:1004 available 996:1004 available 996:1004 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 5 25 theme executable 684:693 arg1 files 695:699 The executable files 680:699 The executable files 680:699 AVAILABILITY AND IMPLEMENTATION The executable files of Mass++ are available for free at http://www.first-ms3d.jp/english/. 25712693 7 26 theme @ 927:927 arg1 shimadzu.co.jp 928:941 acyshzw@shimadzu.co.jp 920:941 acyshzw@shimadzu.co.jp 920:941 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 2 27 theme glycan 286:291 arg1 structures 293:302 glycan structures 286:302 glycan structures from MS/MS spectra 286:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 1 28 theme potent 154:159 arg1 technique 161:169 a potent technique 152:169 a potent technique for characterizing N-glycan structures 152:208 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 28 theme potent 154:159 arg1 spectrometry 119:130 UNLABELLED Tandem mass spectrometry 96:130 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) 96:147 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 3 29 theme glycan 362:367 arg1 structures 369:378 glycan structures 362:378 glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) 362:437 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25712693 1 30 theme Tandem 107:112 arg1 technique 161:169 a potent technique 152:169 a potent technique for characterizing N-glycan structures 152:208 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 30 theme Tandem 107:112 arg1 spectrometry 119:130 UNLABELLED Tandem mass spectrometry 96:130 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) 96:147 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 4 31 theme freeware 589:596 arg1 spectrum 603:610 a freeware mass spectrum 587:610 a freeware mass spectrum visualization 587:624 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 0 32 theme search 35:40 arg1 tool 42:45 a database search tool 24:45 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 0:94 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 0 33 theme database 26:33 arg1 tool 42:45 a database search tool 24:45 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 0:94 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 5 34 theme Mass++ 704:709 arg1 IMPLEMENTATION 665:678 IMPLEMENTATION 665:678 IMPLEMENTATION 665:678 AVAILABILITY AND IMPLEMENTATION The executable files of Mass++ are available for free at http://www.first-ms3d.jp/english/. 25712693 5 34 theme Mass++ 704:709 arg1 AVAILABILITY 648:659 AVAILABILITY 648:659 AVAILABILITY 648:659 AVAILABILITY AND IMPLEMENTATION The executable files of Mass++ are available for free at http://www.first-ms3d.jp/english/. 25712693 4 35 theme spectrum 603:610 arg1 Mass++ 579:584 Mass++ 579:584 Mass++ 579:584 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 4 35 theme spectrum 603:610 arg1 visualization 612:624 a freeware mass spectrum visualization 587:624 a freeware mass spectrum visualization 587:624 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 1 36 theme mass 114:117 arg1 technique 161:169 a potent technique 152:169 a potent technique for characterizing N-glycan structures 152:208 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 36 theme mass 114:117 arg1 spectrometry 119:130 UNLABELLED Tandem mass spectrometry 96:130 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) 96:147 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 7 37 theme SUPPLEMENTARY 943:955 arg1 material 983:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 SUPPLEMENTARY INFORMATION Supplementary material 943:990 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 6 38 theme Mass++ 838:843 arg1 package 827:833 the standard package 814:833 the standard package of Mass++ for Windows 814:855 The GlycanAnalysis plug-in is included in the standard package of Mass++ for Windows. 25712693 1 39 theme UNLABELLED 96:105 arg1 technique 161:169 a potent technique 152:169 a potent technique for characterizing N-glycan structures 152:208 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 39 theme UNLABELLED 96:105 arg1 spectrometry 119:130 UNLABELLED Tandem mass spectrometry 96:130 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) 96:147 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 1 40 theme N-glycan 190:197 arg1 structures 199:208 N-glycan structures 190:208 N-glycan structures 190:208 UNLABELLED Tandem mass spectrometry (MS/MS or MS(n)) is a potent technique for characterizing N-glycan structures. 25712693 2 41 theme glycan 237:242 arg1 database 244:251 a glycan database 235:251 a glycan database to support the identification of glycan structures from MS/MS spectra 235:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 7 42 theme Bioinformatics 1009:1022 arg1 online 1024:1029 Bioinformatics online 1009:1029 Bioinformatics online 1009:1029 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 2 43 theme MS/MS 309:313 arg1 spectra 315:321 MS/MS spectra 309:321 MS/MS spectra 309:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 2 44 from identification 268:281 arg1 spectra 315:321 MS/MS spectra 309:321 MS/MS spectra 309:321 GlycanAnalysis searches a glycan database to support the identification of glycan structures from MS/MS spectra. 25712693 5 45 from http 737:740 arg1 available 715:723 available 715:723 available 715:723 AVAILABILITY AND IMPLEMENTATION The executable files of Mass++ are available for free at http://www.first-ms3d.jp/english/. 25712693 0 46 dep Plug-in 15:21 arg1 tool 42:45 a database search tool 24:45 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 0:94 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 4 47 theme mass 598:601 arg1 spectrum 603:610 a freeware mass spectrum 587:610 a freeware mass spectrum visualization 587:624 This program functions as a plug-in for Mass++, a freeware mass spectrum visualization and analysis program. 25712693 7 48 theme @ 901:901 arg1 shimadzu.co.jp 902:915 nishikaz@shimadzu.co.jp 893:915 nishikaz@shimadzu.co.jp 893:915 CONTACT k-morimt@shimadzu.co.jp or nishikaz@shimadzu.co.jp or acyshzw@shimadzu.co.jp SUPPLEMENTARY INFORMATION Supplementary material are available at Bioinformatics online. 25712693 0 49 theme N-glycan 51:58 arg1 structures 60:69 N-glycan structures 51:69 N-glycan structures using mass spectrometry 51:93 GlycanAnalysis Plug-in: a database search tool for N-glycan structures using mass spectrometry. 25712693 3 50 theme diagnostic 494:503 arg1 ions 505:508 diagnostic ions 494:508 diagnostic ions 494:508 It also calculates diagnostic ions of glycan structures registered in a glycan database (GlycomeDB or KEGG GLYCAN) and searches for MS/MS spectra of N-glycans that match diagnostic ions to determine the structures. 25358049 5 0 gly glycopeptides 1112:1124 arg2 glycopeptides 1112:1124 such glycopeptides 1107:1124 such glycopeptides 1107:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 4 1 gly glycopeptides 912:924 arg2 glycopeptides 912:924 glycopeptides 912:924 glycopeptides 912:924 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 4 2 theme Galβ3GlcNAcβ-O- 1019:1033 arg1 structures 1035:1044 Galβ3GlcNAcβ-O- structures 1019:1044 Galβ3GlcNAcβ-O- structures 1019:1044 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 0 3 theme profiles 82:89 arg1 analysis 44:51 analysis 44:51 analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides 44:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 5 4 theme such 1107:1110 arg1 glycopeptides 1112:1124 such glycopeptides 1107:1124 such glycopeptides 1107:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 5 5 theme glycopeptides 1112:1124 arg1 profiles 1095:1102 the oxonium ion fragmentation profiles 1065:1102 the oxonium ion fragmentation profiles of such glycopeptides 1065:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 5 6 from importance 1204:1213 arg1 studies 1248:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 4 7 theme oxonium 865:871 arg1 ions 873:876 oxonium ions 865:876 oxonium ions 865:876 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 8 theme oxonium 620:626 arg1 ions 628:631 saccharide oxonium ions 609:631 saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues 609:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 5 9 theme fragmentation 1081:1093 arg1 profiles 1095:1102 the oxonium ion fragmentation profiles 1065:1102 the oxonium ion fragmentation profiles of such glycopeptides 1065:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 5 10 theme LC-MS/MS-based 1218:1231 arg1 studies 1248:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 5 11 used used 1138:1141 arg2 difference 1051:1060 The difference 1047:1060 The difference in the oxonium ion fragmentation profiles of such glycopeptides 1047:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 4 12 theme O-glycopeptides 804:818 arg1 spectra 730:736 beam-type CID spectra 716:736 beam-type CID spectra 716:736 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 4 12 theme O-glycopeptides 804:818 arg1 spectra 755:761 ion trap CID spectra 742:761 ion trap CID spectra 742:761 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 4 13 theme N- 797:798 arg1 spectra 730:736 beam-type CID spectra 716:736 beam-type CID spectra 716:736 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 4 13 theme N- 797:798 arg1 spectra 755:761 ion trap CID spectra 742:761 ion trap CID spectra 742:761 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 14 theme liquid 507:512 arg1 LC-MS/MS 555:562 LC-MS/MS 555:562 LC-MS/MS 555:562 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 14 theme liquid 507:512 arg1 spectrometry 541:552 liquid chromatography-tandem mass spectrometry 507:552 liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 507:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 15 theme glycoproteomic 478:491 arg1 analysis 493:500 glycoproteomic analysis 478:500 glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 478:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 4 16 theme trap 746:749 arg1 spectra 755:761 ion trap CID spectra 742:761 ion trap CID spectra 742:761 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 0 17 theme glycopeptides 106:118 arg1 LC-MS/MS 94:101 LC-MS/MS 94:101 LC-MS/MS of glycopeptides 94:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 0 18 from analysis 44:51 arg1 LC-MS/MS 94:101 LC-MS/MS 94:101 LC-MS/MS of glycopeptides 94:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 5 19 from difference 1051:1060 arg1 profiles 1095:1102 the oxonium ion fragmentation profiles 1065:1102 the oxonium ion fragmentation profiles of such glycopeptides 1065:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 3 20 theme saccharide 609:618 arg1 ions 628:631 saccharide oxonium ions 609:631 saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues 609:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 2 21 theme biosynthetic 436:447 arg1 pathways 449:456 biosynthetic pathways 436:456 biosynthetic pathways being conserved 436:472 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 4 22 theme ion 742:744 arg1 spectra 755:761 ion trap CID spectra 742:761 ion trap CID spectra 742:761 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 2 23 theme glycan 353:358 arg1 characterization 333:348 characterization 333:348 characterization of glycan and glycopeptide structural isomers 333:394 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 5 24 theme glycoproteomic 1233:1246 arg1 studies 1248:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 LC-MS/MS-based glycoproteomic studies 1218:1254 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 1 25 theme biological 193:202 arg1 processes 204:212 diverse biological processes 185:212 diverse biological processes 185:212 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 4 26 link derived 789:795 arg1 N- 797:798 synthetic and natively derived N- 766:798 synthetic and natively derived N- 766:798 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 27 theme HexNAc 664:669 arg1 residues 672:679 N-acetylhexosamine (HexNAc) residues 644:679 N-acetylhexosamine (HexNAc) residues 644:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 1 28 theme processes 204:212 arg1 regulation 171:180 the regulation 167:180 the regulation of diverse biological processes 167:212 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 3 29 contain containing 633:642 arg1 ions 628:631 saccharide oxonium ions 609:631 saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues 609:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 29 contain containing 633:642 arg2 residues 672:679 N-acetylhexosamine (HexNAc) residues 644:679 N-acetylhexosamine (HexNAc) residues 644:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 0 30 gly glycopeptides 106:118 arg2 glycopeptides 106:118 glycopeptides 106:118 glycopeptides 106:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 2 31 dep glycan 353:358 arg1 isomers 388:394 structural isomers 377:394 structural isomers 377:394 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 4 32 theme CID 726:728 arg1 spectra 730:736 beam-type CID spectra 716:736 beam-type CID spectra 716:736 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 0 33 theme saccharide 14:23 arg1 Assignment 0:9 Assignment 0:9 Assignment of saccharide 0:23 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 5 34 theme oxonium 1069:1075 arg1 profiles 1095:1102 the oxonium ion fragmentation profiles 1065:1102 the oxonium ion fragmentation profiles of such glycopeptides 1065:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 3 35 theme N-acetylhexosamine 644:661 arg1 residues 672:679 N-acetylhexosamine (HexNAc) residues 644:679 N-acetylhexosamine (HexNAc) residues 644:679 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 4 36 theme beam-type 716:724 arg1 spectra 730:736 beam-type CID spectra 716:736 beam-type CID spectra 716:736 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 37 theme collision-induced 571:587 arg1 CID 603:605 CID 603:605 CID 603:605 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 37 theme collision-induced 571:587 arg1 dissociation 589:600 collision-induced dissociation 571:600 collision-induced dissociation (CID) 571:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 0 38 from profiles 82:89 arg1 LC-MS/MS 94:101 LC-MS/MS 94:101 LC-MS/MS of glycopeptides 94:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 4 39 theme spectra 730:736 arg1 analysis 704:711 analysis 704:711 analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides 704:818 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 40 from prominent 685:693 arg1 analysis 493:500 glycoproteomic analysis 478:500 glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 478:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 41 theme chromatography-tandem 514:534 arg1 LC-MS/MS 555:562 LC-MS/MS 555:562 LC-MS/MS 555:562 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 41 theme chromatography-tandem 514:534 arg1 spectrometry 541:552 liquid chromatography-tandem mass spectrometry 507:552 liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 507:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 4 42 gly O-glycopeptides 804:818 arg2 O-glycopeptides 804:818 O-glycopeptides 804:818 O-glycopeptides 804:818 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 3 43 theme mass 536:539 arg1 LC-MS/MS 555:562 LC-MS/MS 555:562 LC-MS/MS 555:562 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 43 theme mass 536:539 arg1 spectrometry 541:552 liquid chromatography-tandem mass spectrometry 507:552 liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 507:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 0 44 theme oxonium 56:62 arg1 profiles 82:89 oxonium ion fragmentation profiles 56:89 oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides 56:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 4 45 theme spectra 755:761 arg1 analysis 704:711 analysis 704:711 analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides 704:818 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 1 46 theme Protein 121:127 arg1 glycosylation 129:141 Protein glycosylation 121:141 Protein glycosylation 121:141 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 3 47 with analysis 493:500 arg1 LC-MS/MS 555:562 LC-MS/MS 555:562 LC-MS/MS 555:562 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 3 47 with analysis 493:500 arg1 spectrometry 541:552 liquid chromatography-tandem mass spectrometry 507:552 liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID) 507:606 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 5 48 theme glycan 1170:1175 arg1 structures 1177:1186 these glycan structures 1164:1186 these glycan structures 1164:1186 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 2 49 theme structural 377:386 arg1 isomers 388:394 structural isomers 377:394 structural isomers 377:394 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 4 50 theme CID 751:753 arg1 spectra 755:761 ion trap CID spectra 742:761 ion trap CID spectra 742:761 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 4 51 theme ions 873:876 arg1 patterns 853:860 the fragmentation patterns 835:860 the fragmentation patterns of oxonium ions 835:876 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 2 52 theme glycopeptide 364:375 arg1 characterization 333:348 characterization 333:348 characterization of glycan and glycopeptide structural isomers 333:394 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 5 53 theme ion 1077:1079 arg1 profiles 1095:1102 the oxonium ion fragmentation profiles 1065:1102 the oxonium ion fragmentation profiles of such glycopeptides 1065:1124 The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies. 25358049 1 54 theme glycan 236:241 arg1 relationships 262:274 glycan structure-function relationships 236:274 glycan structure-function relationships 236:274 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 2 55 gly glycopeptide 364:375 arg2 glycopeptide 364:375 glycopeptide 364:375 glycopeptide 364:375 However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. 25358049 4 56 theme fragmentation 839:851 arg1 patterns 853:860 the fragmentation patterns 835:860 the fragmentation patterns of oxonium ions 835:876 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 1 57 theme critical 149:156 arg1 roles 158:162 critical roles 149:162 critical roles 149:162 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 1 58 theme structure-function 243:260 arg1 relationships 262:274 glycan structure-function relationships 236:274 glycan structure-function relationships 236:274 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 0 59 theme fragmentation 68:80 arg1 profiles 82:89 oxonium ion fragmentation profiles 56:89 oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides 56:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 0 60 from LC-MS/MS 94:101 arg1 analysis 44:51 analysis 44:51 analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides 44:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 3 61 from analysis 493:500 arg1 prominent 685:693 prominent 685:693 prominent 685:693 In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. 25358049 4 62 theme synthetic 766:774 arg1 N- 797:798 synthetic and natively derived N- 766:798 synthetic and natively derived N- 766:798 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 25358049 1 63 theme diverse 185:191 arg1 processes 204:212 diverse biological processes 185:212 diverse biological processes 185:212 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 1 64 theme relationships 262:274 arg1 determination 219:231 determination 219:231 determination of glycan structure-function relationships 219:274 Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. 25358049 0 65 theme ion 64:66 arg1 profiles 82:89 oxonium ion fragmentation profiles 56:89 oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides 56:118 Assignment of saccharide identities through analysis of oxonium ion fragmentation profiles in LC-MS/MS of glycopeptides. 25358049 4 66 theme derived 789:795 arg1 N- 797:798 synthetic and natively derived N- 766:798 synthetic and natively derived N- 766:798 Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAcα1-O-, GlcNAcβ1-O-, Galβ3GalNAcα1-O-, Galβ4GlcNAcβ-O-, and Galβ3GlcNAcβ-O- structures. 29125207 2 0 theme β-mannose 562:570 arg1 moiety 572:577 the β-mannose moiety 558:577 the β-mannose moiety 558:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 5 1 dep results 899:905 arg1 the 880:882 the 880:882 the 880:882 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 5 1 dep results 899:905 arg1 basis 884:888 basis 884:888 basis 884:888 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 1 2 theme oxazoline 305:313 arg1 derivatives 315:325 tetrasaccharide oxazoline derivatives 289:325 tetrasaccharide oxazoline derivatives 289:325 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 2 3 theme glycan 432:437 arg1 structure 385:393 the core structure 376:393 the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 376:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 3 4 dep wildtype 615:622 arg1 WT 625:626 WT 625:626 WT 625:626 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 1 5 theme derivatives 315:325 arg1 series 279:284 a series 277:284 a series of tetrasaccharide oxazoline derivatives 277:325 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 2 6 link asparagine-linked 401:417 arg1 glycan 432:437 an asparagine-linked glycoprotein glycan 398:437 an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 398:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 4 7 theme group 819:823 arg1 configuration 790:802 The essential configuration 776:802 The essential configuration of the hydroxy group for the transglycosylation reaction 776:859 The essential configuration of the hydroxy group for the transglycosylation reaction was determined. 29125207 4 8 theme transglycosylation 833:850 arg1 reaction 852:859 the transglycosylation reaction 829:859 the transglycosylation reaction 829:859 The essential configuration of the hydroxy group for the transglycosylation reaction was determined. 29125207 1 9 theme glycosyl 156:163 arg1 donor 165:169 the glycosyl donor 152:169 the glycosyl donor 152:169 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 5 10 theme transglycosylation 912:929 arg1 reaction 931:938 the transglycosylation reaction 908:938 the transglycosylation reaction 908:938 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 3 11 theme wildtype 615:622 arg1 endo-M 629:634 wildtype (WT) endo-M 615:634 wildtype (WT) endo-M 615:634 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 2 12 theme non-natural-type 466:481 arg1 monosaccharide 483:496 a non-natural-type monosaccharide 464:496 a non-natural-type monosaccharide 464:496 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 1 13 theme structural 126:135 arg1 specificity 137:147 the structural specificity 122:147 the structural specificity of the glycosyl donor for the transglycosylation reaction 122:205 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 1 14 theme Mucor 252:256 arg1 endo-M 268:273 endo-M 268:273 endo-M 268:273 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 1 14 theme Mucor 252:256 arg1 hiemalis 258:265 Mucor hiemalis 252:265 Mucor hiemalis (endo-M) 252:274 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 2 15 theme core 380:383 arg1 structure 385:393 the core structure 376:393 the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 376:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 0 16 theme Structures 21:30 arg1 Specificity 0:10 Specificity 0:10 Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions. 0:105 Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions. 29125207 4 17 theme hydroxy 811:817 arg1 group 819:823 the hydroxy group 807:823 the hydroxy group 807:823 The essential configuration of the hydroxy group for the transglycosylation reaction was determined. 29125207 3 18 theme endo-M 629:634 arg1 activity 603:610 The transglycosylation activity 580:610 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, 580:668 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 0 19 theme Donor 15:19 arg1 Structures 21:30 Donor Structures 15:30 Donor Structures 15:30 Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions. 29125207 5 20 gly transglycosylated 998:1014 arg1 products 1016:1023 transglycosylated products 998:1023 transglycosylated products 998:1023 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 3 21 theme tetrasaccharide 698:712 arg1 donors 714:719 these tetrasaccharide donors 692:719 these tetrasaccharide donors 692:719 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 3 22 theme transglycosylation 584:601 arg1 activity 603:610 The transglycosylation activity 580:610 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, 580:668 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 1 23 theme transglycosylation 179:196 arg1 reaction 198:205 the transglycosylation reaction 175:205 the transglycosylation reaction 175:205 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 0 24 theme endo-β-N-Acetylglucosaminidase-Catalyzed 36:75 arg1 Reactions 96:104 endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions 36:104 endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions 36:104 Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions. 29125207 2 25 theme asparagine-linked 401:417 arg1 glycan 432:437 an asparagine-linked glycoprotein glycan 398:437 an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 398:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 4 26 theme essential 780:788 arg1 configuration 790:802 The essential configuration 776:802 The essential configuration of the hydroxy group for the transglycosylation reaction 776:859 The essential configuration of the hydroxy group for the transglycosylation reaction was determined. 29125207 2 27 theme monosaccharide 483:496 arg1 unit 456:459 a β-mannose unit 444:459 a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 444:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 5 28 theme modified 977:984 arg1 donors 986:991 chemically modified donors 966:991 chemically modified donors 966:991 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 2 29 theme glycoprotein 419:430 arg1 glycan 432:437 an asparagine-linked glycoprotein glycan 398:437 an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 398:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 0 30 theme Transglycosylation 77:94 arg1 Reactions 96:104 endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions 36:104 endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions 36:104 Specificity of Donor Structures for endo-β-N-Acetylglucosaminidase-Catalyzed Transglycosylation Reactions. 29125207 2 31 gly glycoprotein 419:430 arg1 glycoprotein 419:430 an asparagine-linked glycoprotein glycan 398:437 an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 398:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 1 32 theme donor 165:169 arg1 specificity 137:147 the structural specificity 122:147 the structural specificity of the glycosyl donor for the transglycosylation reaction 122:205 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 2 33 with glycan 432:437 arg1 unit 456:459 a β-mannose unit 444:459 a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 444:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 5 34 theme transglycosylated 998:1014 arg1 products 1016:1023 transglycosylated products 998:1023 transglycosylated products 998:1023 On the basis of these results, the transglycosylation reaction was investigated by using chemically modified donors, and transglycosylated products were successfully obtained. 29125207 3 35 theme p-nitrophenyl 726:738 arg1 GlcNAc-pNp 763:772 GlcNAc-pNp 763:772 GlcNAc-pNp 763:772 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 3 35 theme p-nitrophenyl 726:738 arg1 N-acetylglucosaminide 740:760 p-nitrophenyl N-acetylglucosaminide 726:760 p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp) 726:773 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 1 36 from hiemalis 258:265 arg1 endo-β-N-acetylglucosaminidase 216:245 endo-β-N-acetylglucosaminidase 216:245 endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M) 216:274 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 29125207 2 37 theme β-mannose 446:454 arg1 unit 456:459 a β-mannose unit 444:459 a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety 444:577 These derivatives correspond to the core structure of an asparagine-linked glycoprotein glycan with a β-mannose unit of a non-natural-type monosaccharide, including β-glucose, β-galactose, and β-talose in place of the β-mannose moiety. 29125207 3 38 theme mutants 644:650 arg1 activity 603:610 The transglycosylation activity 580:610 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, 580:668 The transglycosylation activity of wildtype (WT) endo-M and two mutants, N175Q and N175A, was examined by using these tetrasaccharide donors with p-nitrophenyl N-acetylglucosaminide (GlcNAc-pNp). 29125207 1 39 theme tetrasaccharide 289:303 arg1 derivatives 315:325 tetrasaccharide oxazoline derivatives 289:325 tetrasaccharide oxazoline derivatives 289:325 To demonstrate the structural specificity of the glycosyl donor for the transglycosylation reaction by using endo-β-N-acetylglucosaminidase from Mucor hiemalis (endo-M), a series of tetrasaccharide oxazoline derivatives was synthesized. 24012618 5 0 theme chromatography 785:798 arg1 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 6 1 theme significant 925:935 arg1 correlation 946:956 A small but significant positive correlation 913:956 A small but significant positive correlation between BMI 913:968 RESULTS A small but significant positive correlation between BMI and the level of neutral glycans without galactoses was detected. 24012618 5 2 from total 878:882 arg1 samples 863:869 plasma samples 856:869 plasma samples from a total of 3515 individuals 856:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 2 from total 878:882 arg1 IgG 849:851 IgG 849:851 IgG in plasma samples from a total of 3515 individuals 849:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 8 3 theme glycans 1316:1322 arg1 level 1299:1303 the level 1295:1303 the level of neutral glycans with two terminal galactoses 1295:1351 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 7 4 from % 1168:1168 arg1 level 1190:1194 the level 1186:1194 the level of neutral glycans without galactoses 1186:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 0 5 theme index 74:78 arg1 galactosylation 24:38 galactosylation 24:38 galactosylation of immunoglobulin G and body mass index 24:78 The association between galactosylation of immunoglobulin G and body mass index. 24012618 9 6 theme IgG 1610:1612 arg1 galactosylation 1614:1628 IgG galactosylation 1610:1628 IgG galactosylation 1610:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 5 7 attach attached 837:844 arg2 N-glycans 827:835 N-glycans 827:835 N-glycans attached to IgG in plasma samples from a total of 3515 individuals 827:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 7 attach attached 837:844 arg1 IgG 849:851 IgG 849:851 IgG in plasma samples from a total of 3515 individuals 849:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 8 from chromatography 749:762 arg1 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 8 9 theme variation 1474:1482 arg1 variation 1474:1482 variation 1474:1482 variation in this glycan feature 1474:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 9 theme variation 1474:1482 arg1 % 1469:1469 up to 3.2% 1460:1469 up to 3.2% of variation in this glycan feature 1460:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 5 10 theme interaction 737:747 arg1 chromatography 749:762 A hydrophilic interaction chromatography 723:762 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column 723:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 6 11 theme small 915:919 arg1 correlation 946:956 A small but significant positive correlation 913:956 A small but significant positive correlation between BMI 913:968 RESULTS A small but significant positive correlation between BMI and the level of neutral glycans without galactoses was detected. 24012618 7 12 theme glycans 1207:1213 arg1 level 1190:1194 the level 1186:1194 the level of neutral glycans without galactoses 1186:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 7 13 from level 1190:1194 arg1 variation 1173:1181 variation 1173:1181 variation in the level of neutral glycans without galactoses 1173:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 7 13 from level 1190:1194 arg1 % 1168:1168 up to 2.0% 1159:1168 up to 2.0% of variation in the level of neutral glycans without galactoses 1159:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 7 14 theme neutral 1199:1205 arg1 glycans 1207:1213 neutral glycans 1199:1213 neutral glycans without galactoses 1199:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 4 15 dep METHOD 577:582 arg1 included 594:601 included 594:601 included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands 594:720 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 10 16 theme increased 1687:1695 arg1 BMI 1697:1699 increased BMI 1687:1699 increased BMI 1687:1699 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 2 17 gly Glycosylation 152:164 arg1 proteins 169:176 proteins 169:176 proteins 169:176 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 6 18 theme glycans 995:1001 arg1 level 978:982 the level 974:982 the level of neutral glycans without galactoses 974:1020 RESULTS A small but significant positive correlation between BMI and the level of neutral glycans without galactoses was detected. 24012618 3 19 theme mass 393:396 arg1 index 398:402 body mass index 388:402 body mass index (BMI) in both Croatian and Chinese populations 388:449 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 3 19 theme mass 393:396 arg1 BMI 405:407 BMI 405:407 BMI 405:407 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 5 20 from IgG 849:851 arg1 samples 863:869 plasma samples 856:869 plasma samples from a total of 3515 individuals 856:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 20 from IgG 849:851 arg1 total 878:882 a total 876:882 a total of 3515 individuals 876:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 21 theme plasma 856:861 arg1 samples 863:869 plasma samples 856:869 plasma samples from a total of 3515 individuals 856:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 2 22 theme cellular 312:319 arg1 processes 321:329 numerous physiological and pathophysiological cellular processes 266:329 numerous physiological and pathophysiological cellular processes 266:329 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 9 23 from changes 1599:1605 arg1 galactosylation 1614:1628 IgG galactosylation 1610:1628 IgG galactosylation 1610:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 10 24 theme observed 1635:1642 arg1 loss 1644:1647 The observed loss 1631:1647 The observed loss of galactose which is associated with increased BMI 1631:1699 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 10 24 theme observed 1635:1642 arg1 related 1710:1716 related 1710:1716 related 1710:1716 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 0 25 theme G 58:58 arg1 galactosylation 24:38 galactosylation 24:38 galactosylation of immunoglobulin G and body mass index 24:78 The association between galactosylation of immunoglobulin G and body mass index. 24012618 7 26 from variation 1173:1181 arg1 level 1190:1194 the level 1186:1194 the level of neutral glycans without galactoses 1186:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 8 27 theme terminal 1333:1340 arg1 galactoses 1342:1351 two terminal galactoses 1329:1351 two terminal galactoses 1329:1351 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 28 theme glycan 1492:1497 arg1 feature 1499:1505 this glycan feature 1487:1505 this glycan feature 1487:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 2 29 theme pathophysiological 293:310 arg1 processes 321:329 numerous physiological and pathophysiological cellular processes 266:329 numerous physiological and pathophysiological cellular processes 266:329 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 7 30 theme variation 1173:1181 arg1 variation 1173:1181 variation 1173:1181 variation in the level of neutral glycans without galactoses 1173:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 7 30 theme variation 1173:1181 arg1 % 1168:1168 up to 2.0% 1159:1168 up to 2.0% of variation in the level of neutral glycans without galactoses 1159:1232 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 5 31 theme BEH 774:776 arg1 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 1 32 theme OBJECTIVE 81:89 arg1 Obesity 91:97 OBJECTIVE Obesity 81:97 OBJECTIVE Obesity 81:97 OBJECTIVE Obesity is becoming a fast-growing health problem worldwide. 24012618 8 33 from BMI 1384:1386 arg1 groups 1407:1412 analyzed sample groups 1391:1412 analyzed sample groups 1391:1412 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 34 from variation 1474:1482 arg1 feature 1499:1505 this glycan feature 1487:1505 this glycan feature 1487:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 6 35 theme neutral 987:993 arg1 glycans 995:1001 neutral glycans 987:1001 neutral glycans without galactoses 987:1020 RESULTS A small but significant positive correlation between BMI and the level of neutral glycans without galactoses was detected. 24012618 7 36 theme correlation 1095:1105 arg1 coefficients 1107:1118 correlation coefficients 1095:1118 correlation coefficients 1095:1118 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 10 37 theme galactose 1652:1660 arg1 loss 1644:1647 The observed loss 1631:1647 The observed loss of galactose which is associated with increased BMI 1631:1699 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 10 37 theme galactose 1652:1660 arg1 related 1710:1716 related 1710:1716 related 1710:1716 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 9 38 theme first 1536:1540 arg1 study 1523:1527 Our study 1519:1527 Our study 1519:1527 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 9 38 theme first 1536:1540 arg1 study 1554:1558 the first large-scale study 1532:1558 the first large-scale study to indicate the association of BMI and changes in IgG galactosylation 1532:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 3 39 from index 398:402 arg1 Croatian 418:425 Croatian 418:425 Croatian 418:425 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 3 39 from index 398:402 arg1 populations 439:449 Chinese populations 431:449 Chinese populations 431:449 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 10 40 theme chronic 1721:1727 arg1 inflammation 1729:1740 chronic inflammation 1721:1740 chronic inflammation that accompanies the development of obesity 1721:1784 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 9 41 theme large-scale 1542:1552 arg1 study 1523:1527 Our study 1519:1527 Our study 1519:1527 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 9 41 theme large-scale 1542:1552 arg1 study 1554:1558 the first large-scale study 1532:1558 the first large-scale study to indicate the association of BMI and changes in IgG galactosylation 1532:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 9 42 from association 1576:1586 arg1 galactosylation 1614:1628 IgG galactosylation 1610:1628 IgG galactosylation 1610:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 3 43 theme immunoglobulin 496:509 arg1 IgG 514:516 IgG 514:516 IgG 514:516 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 3 43 theme immunoglobulin 496:509 arg1 G 511:511 immunoglobulin G 496:511 immunoglobulin G (IgG) glycome 496:525 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 3 44 theme body 388:391 arg1 index 398:402 body mass index 388:402 body mass index (BMI) in both Croatian and Chinese populations 388:449 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 3 44 theme body 388:391 arg1 BMI 405:407 BMI 405:407 BMI 405:407 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 7 45 theme gender 1087:1092 arg1 influence 1066:1074 the influence 1062:1074 the influence of age and gender 1062:1092 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 8 46 from feature 1499:1505 arg1 variation 1474:1482 variation 1474:1482 variation in this glycan feature 1474:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 46 from feature 1499:1505 arg1 % 1469:1469 up to 3.2% 1460:1469 up to 3.2% of variation in this glycan feature 1460:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 47 theme neutral 1308:1314 arg1 glycans 1316:1322 neutral glycans 1308:1322 neutral glycans with two terminal galactoses 1308:1351 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 3 48 theme G 511:511 arg1 glycome 519:525 immunoglobulin G (IgG) glycome 496:525 immunoglobulin G (IgG) glycome 496:525 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 0 49 theme immunoglobulin 43:56 arg1 G 58:58 immunoglobulin G 43:58 immunoglobulin G 43:58 The association between galactosylation of immunoglobulin G and body mass index. 24012618 5 50 used used 811:814 arg2 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 5 50 used used 811:814 arg2 chromatography 749:762 A hydrophilic interaction chromatography 723:762 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column 723:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 3 51 theme biological 547:556 arg1 function 558:565 biological function 547:565 biological function 547:565 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 4 52 from islands 642:648 arg1 individuals 603:613 individuals 603:613 individuals from two Croatian Adriatic islands, Vis and Korčula, 603:666 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 52 from islands 642:648 arg1 individuals 672:682 individuals 672:682 individuals from Northern Scottish Orkney Islands 672:720 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 8 53 theme age 1279:1281 arg1 effects 1268:1274 the effects 1264:1274 the effects of age and gender 1264:1292 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 7 54 dep 2.0 1165:1167 arg1 to 1162:1163 to 1162:1163 to 1162:1163 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 5 55 theme hydrophilic 725:735 arg1 chromatography 749:762 A hydrophilic interaction chromatography 723:762 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column 723:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 10 56 theme obesity 1778:1784 arg1 development 1763:1773 the development 1759:1773 the development of obesity 1759:1784 The observed loss of galactose which is associated with increased BMI might be related to chronic inflammation that accompanies the development of obesity. 24012618 5 57 theme Waters 767:772 arg1 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 8 58 theme gender 1287:1292 arg1 effects 1268:1274 the effects 1264:1274 the effects of age and gender 1264:1292 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 2 59 theme proteins 169:176 arg1 variations 188:197 their variations 182:197 their variations 182:197 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 2 59 theme proteins 169:176 arg1 Glycosylation 152:164 Glycosylation 152:164 Glycosylation of proteins 152:176 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 8 60 theme sample 1400:1405 arg1 groups 1407:1412 analyzed sample groups 1391:1412 analyzed sample groups 1391:1412 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 7 61 theme age 1079:1081 arg1 influence 1066:1074 the influence 1062:1074 the influence of age and gender 1062:1092 After taking into account the influence of age and gender, correlation coefficients indicated that BMI was responsible for up to 2.0% of variation in the level of neutral glycans without galactoses. 24012618 3 62 theme Chinese 431:437 arg1 populations 439:449 Chinese populations 431:449 Chinese populations 431:449 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 8 63 with glycans 1316:1322 arg1 galactoses 1342:1351 two terminal galactoses 1329:1351 two terminal galactoses 1329:1351 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 8 64 theme analyzed 1391:1398 arg1 groups 1407:1412 analyzed sample groups 1391:1412 analyzed sample groups 1391:1412 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 1 65 theme fast-growing 113:124 arg1 worldwide 141:149 a fast-growing health problem worldwide 111:149 a fast-growing health problem worldwide 111:149 OBJECTIVE Obesity is becoming a fast-growing health problem worldwide. 24012618 8 66 dep 3.2 1466:1468 arg1 to 1463:1464 to 1463:1464 to 1463:1464 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 2 67 theme protein 220:226 arg1 structure 228:236 protein structure 220:236 protein structure 220:236 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 5 68 theme Glycan 778:783 arg1 column 800:805 Waters BEH Glycan chromatography column 767:805 Waters BEH Glycan chromatography column 767:805 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 1 69 theme health 126:131 arg1 worldwide 141:149 a fast-growing health problem worldwide 111:149 a fast-growing health problem worldwide 111:149 OBJECTIVE Obesity is becoming a fast-growing health problem worldwide. 24012618 9 70 theme BMI 1591:1593 arg1 association 1576:1586 the association 1572:1586 the association of BMI 1572:1593 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 9 70 theme BMI 1591:1593 arg1 changes 1599:1605 changes 1599:1605 changes in IgG galactosylation 1599:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 3 71 theme plasma 338:343 arg1 glycans 345:351 plasma glycans 338:351 plasma glycans 338:351 Since plasma glycans were significantly associated with body mass index (BMI) in both Croatian and Chinese populations, the study evaluated the association between immunoglobulin G (IgG) glycome, which is closer to biological function, and BMI. 24012618 2 72 theme physiological 275:287 arg1 processes 321:329 numerous physiological and pathophysiological cellular processes 266:329 numerous physiological and pathophysiological cellular processes 266:329 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 1 73 theme problem 133:139 arg1 worldwide 141:149 a fast-growing health problem worldwide 111:149 a fast-growing health problem worldwide 111:149 OBJECTIVE Obesity is becoming a fast-growing health problem worldwide. 24012618 9 74 dep CONCLUSION 1508:1517 arg1 study 1523:1527 Our study 1519:1527 Our study 1519:1527 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 9 74 dep CONCLUSION 1508:1517 arg1 study 1554:1558 the first large-scale study 1532:1558 the first large-scale study to indicate the association of BMI and changes in IgG galactosylation 1532:1628 CONCLUSION Our study is the first large-scale study to indicate the association of BMI and changes in IgG galactosylation. 24012618 0 75 theme mass 69:72 arg1 index 74:78 body mass index 64:78 body mass index 64:78 The association between galactosylation of immunoglobulin G and body mass index. 24012618 2 76 theme numerous 266:273 arg1 processes 321:329 numerous physiological and pathophysiological cellular processes 266:329 numerous physiological and pathophysiological cellular processes 266:329 Glycosylation of proteins and their variations significantly affect protein structure and function, thus altering numerous physiological and pathophysiological cellular processes. 24012618 5 77 theme individuals 892:902 arg1 total 878:882 a total 876:882 a total of 3515 individuals 876:902 A hydrophilic interaction chromatography on Waters BEH Glycan chromatography column was used to analyze N-glycans attached to IgG in plasma samples from a total of 3515 individuals. 24012618 4 78 from Islands 714:720 arg1 individuals 603:613 individuals 603:613 individuals from two Croatian Adriatic islands, Vis and Korčula, 603:666 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 78 from Islands 714:720 arg1 individuals 672:682 individuals 672:682 individuals from Northern Scottish Orkney Islands 672:720 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 79 theme Adriatic 633:640 arg1 Vis 651:653 Vis 651:653 Vis 651:653 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 79 theme Adriatic 633:640 arg1 islands 642:648 two Croatian Adriatic islands 620:648 two Croatian Adriatic islands 620:648 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 79 theme Adriatic 633:640 arg1 Korčula 659:665 Korčula 659:665 Korčula 659:665 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 0 80 theme body 64:67 arg1 index 74:78 body mass index 64:78 body mass index 64:78 The association between galactosylation of immunoglobulin G and body mass index. 24012618 6 81 theme positive 937:944 arg1 correlation 946:956 A small but significant positive correlation 913:956 A small but significant positive correlation between BMI 913:968 RESULTS A small but significant positive correlation between BMI and the level of neutral glycans without galactoses was detected. 24012618 8 82 from % 1469:1469 arg1 feature 1499:1505 this glycan feature 1487:1505 this glycan feature 1487:1505 Furthermore, after adjusting the effects of age and gender, the level of neutral glycans with two terminal galactoses was negatively associated with BMI in analyzed sample groups, suggesting that BMI could be responsible for up to 3.2% of variation in this glycan feature. 24012618 4 83 theme Croatian 624:631 arg1 Vis 651:653 Vis 651:653 Vis 651:653 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 83 theme Croatian 624:631 arg1 islands 642:648 two Croatian Adriatic islands 620:648 two Croatian Adriatic islands 620:648 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 24012618 4 83 theme Croatian 624:631 arg1 Korčula 659:665 Korčula 659:665 Korčula 659:665 METHOD The study included individuals from two Croatian Adriatic islands, Vis and Korčula, and individuals from Northern Scottish Orkney Islands. 26034352 7 0 theme normal 1385:1390 arg1 rats 1392:1395 normal rats 1385:1395 normal rats 1385:1395 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 10 1 theme HCC 1914:1916 arg1 tissues 1924:1930 HCC liver tissues 1914:1930 HCC liver tissues 1914:1930 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 2 theme biosynthetic 1601:1612 arg1 pathway 1614:1620 the glycan biosynthetic pathway 1590:1620 the glycan biosynthetic pathway 1590:1620 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 6 3 dep ± 1189:1189 arg1 < 1214:1214 P < 0.01 1212:1219 P < 0.01 1212:1219 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 2 4 used used 415:418 arg2 METHODS 301:307 METHODS 301:307 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; 301:389 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 6 5 theme liver 1256:1260 arg1 tissues 1262:1268 normal liver tissues 1249:1268 normal liver tissues 1249:1268 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 7 6 theme HCC 1351:1353 arg1 rats 1355:1358 HCC rats 1351:1358 HCC rats 1351:1358 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 5 7 theme N-glycomic 875:884 arg1 analysis 886:893 The N-glycomic analysis 871:893 The N-glycomic analysis 871:893 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 10 8 from common 1904:1909 arg1 tissues 1924:1930 HCC liver tissues 1914:1930 HCC liver tissues 1914:1930 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 10 9 theme HCC-associated 1768:1781 arg1 changes 1783:1789 The reported HCC-associated changes 1755:1789 The reported HCC-associated changes in glycan flow and subcellular localization 1755:1833 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 10 theme enzyme 1657:1662 arg1 activities 1664:1673 enzyme activities 1657:1673 enzyme activities involved in glycan synthesis at different subcellular localizations 1657:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 6 11 theme HCC 1225:1227 arg1 tissues 1229:1235 HCC tissues 1225:1235 HCC tissues 1225:1235 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 1 12 from carcinoma 273:281 arg1 rodents 292:298 rodents 292:298 rodents 292:298 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 3 13 theme protein-associated 571:588 arg1 glycans 590:596 protein-associated glycans 571:596 protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 571:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 4 14 theme Histopathological 764:780 arg1 studies 782:788 Histopathological studies 764:788 Histopathological studies 764:788 Histopathological studies were carried out to ensure the development of HCC in the tested animals. 26034352 1 15 theme qualitative 131:141 arg1 changes 160:166 the qualitative and quantitative changes 127:166 the qualitative and quantitative changes 127:166 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 10 16 dep CONCLUSION 1744:1753 arg1 explain 1835:1841 explain 1835:1841 explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues 1835:1930 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 17 theme glycan 1687:1692 arg1 synthesis 1694:1702 glycan synthesis 1687:1702 glycan synthesis at different subcellular localizations 1687:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 7 18 theme P 1311:1311 arg1 < 1313:1313 P < 0.05 1311:1318 P < 0.05 1311:1318 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 3 19 theme unique 606:611 arg1 technology 638:647 a unique, automated 96-well assay technology 604:647 a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 604:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 1 20 theme quantitative 147:158 arg1 changes 160:166 the qualitative and quantitative changes 127:166 the qualitative and quantitative changes 127:166 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 1 21 from changes 160:166 arg1 glycosylation 180:192 N-linked glycosylation 171:192 N-linked glycosylation 171:192 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 9 22 theme subcellular 1717:1727 arg1 localizations 1729:1741 different subcellular localizations 1707:1741 different subcellular localizations 1707:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 2 23 dep rats 357:360 arg1 1 327:327 1 327:327 1 327:327 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 0 24 theme glycomic 92:99 arg1 analysis 101:108 A glycomic analysis 90:108 A glycomic analysis 90:108 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 5 25 theme significant 1030:1040 arg1 changes 1042:1048 significant changes 1030:1048 significant changes in rat HCC tissues 1030:1067 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 2 26 theme tumor-bearing 371:383 arg1 rats 385:388 (2) tumor-bearing rats 367:388 (2) tumor-bearing rats 367:388 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 10 27 from changes 1783:1789 arg1 flow 1801:1804 glycan flow 1794:1804 glycan flow 1794:1804 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 10 27 from changes 1783:1789 arg1 localization 1822:1833 subcellular localization 1810:1833 subcellular localization 1810:1833 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 1 28 link N-linked 171:178 arg1 glycosylation 180:192 N-linked glycosylation 171:192 N-linked glycosylation 171:192 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 6 29 dep increased 1128:1136 arg1 < 1141:1141 P < 0.05 1139:1146 P < 0.05 1139:1146 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 10 30 theme common 1904:1909 arg1 glycans 1872:1878 high mannose glycans 1859:1878 high mannose glycans 1859:1878 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 0 31 theme glycan 14:19 arg1 biosynthesis 21:32 glycan biosynthesis 14:32 glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats 14:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 3 32 theme assay 632:636 arg1 technology 638:647 a unique, automated 96-well assay technology 604:647 a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 604:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 3 33 theme custom 741:746 arg1 bioinformatics 748:761 custom bioinformatics 741:761 custom bioinformatics 741:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 2 34 theme non-tumor-bearing 338:354 arg1 rats 357:360 normal (non-tumor-bearing) rats 330:360 normal (non-tumor-bearing) rats 330:360 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 0 35 theme hepatocellular 56:69 arg1 carcinoma 71:79 chemically-induced hepatocellular carcinoma 37:79 chemically-induced hepatocellular carcinoma in rats 37:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 0 36 from biosynthesis 21:32 arg1 carcinoma 71:79 chemically-induced hepatocellular carcinoma 37:79 chemically-induced hepatocellular carcinoma in rats 37:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 5 37 theme liver 1095:1099 arg1 tissues 1101:1107 normal liver tissues 1088:1107 normal liver tissues 1088:1107 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 3 38 theme desorption/ionization 682:702 arg1 spectrometry 724:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 4 39 theme HCC 836:838 arg1 development 821:831 the development 817:831 the development of HCC in the tested animals 817:860 Histopathological studies were carried out to ensure the development of HCC in the tested animals. 26034352 2 40 theme normal 330:335 arg1 rats 357:360 normal (non-tumor-bearing) rats 330:360 normal (non-tumor-bearing) rats 330:360 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 1 41 theme diethyl 230:236 arg1 HCC 284:286 HCC 284:286 HCC 284:286 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 1 41 theme diethyl 230:236 arg1 carcinoma 273:281 diethyl nitrosamine-induced hepatocellular carcinoma 230:281 diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents 230:298 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 5 42 theme HCC 1057:1059 arg1 tissues 1061:1067 rat HCC tissues 1053:1067 rat HCC tissues 1053:1067 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 3 43 theme matrix-assisted 660:674 arg1 spectrometry 724:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 1 44 theme hepatocellular 258:271 arg1 HCC 284:286 HCC 284:286 HCC 284:286 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 1 44 theme hepatocellular 258:271 arg1 carcinoma 273:281 diethyl nitrosamine-induced hepatocellular carcinoma 230:281 diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents 230:298 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 10 45 theme siayl 1884:1888 arg1 glycans 1896:1902 siayl Lewis glycans 1884:1902 siayl Lewis glycans 1884:1902 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 2 46 theme rats 357:360 arg1 tissues 315:321 Liver tissues 309:321 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; 301:389 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 5 47 dep RESULTS 863:869 arg1 revealed 895:902 revealed 895:902 revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues 895:1107 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 10 48 theme subcellular 1810:1820 arg1 localization 1822:1833 subcellular localization 1810:1833 subcellular localization 1810:1833 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 3 49 theme mass 719:722 arg1 spectrometry 724:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 2 50 theme Liver 309:313 arg1 tissues 315:321 Liver tissues 309:321 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; 301:389 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 10 51 from increase 1847:1854 arg1 glycans 1872:1878 high mannose glycans 1859:1878 high mannose glycans 1859:1878 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 10 51 from increase 1847:1854 arg1 glycans 1896:1902 siayl Lewis glycans 1884:1902 siayl Lewis glycans 1884:1902 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 52 theme glycan 1594:1599 arg1 pathway 1614:1620 the glycan biosynthetic pathway 1590:1620 the glycan biosynthetic pathway 1590:1620 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 1 53 theme N-linked 171:178 arg1 glycosylation 180:192 N-linked glycosylation 171:192 N-linked glycosylation 171:192 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 6 54 theme normal 1249:1254 arg1 tissues 1262:1268 normal liver tissues 1249:1268 normal liver tissues 1249:1268 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 3 55 theme quantitative 547:558 arg1 readout 560:566 a structural and quantitative readout 530:566 a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 530:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 7 56 from increase 1274:1281 arg1 structures 1337:1346 high-mannose structures 1324:1346 high-mannose structures 1324:1346 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 7 56 from increase 1274:1281 arg1 rats 1355:1358 HCC rats 1351:1358 HCC rats 1351:1358 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 3 57 theme laser 676:680 arg1 spectrometry 724:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 660:735 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 3 58 theme structural 532:541 arg1 readout 560:566 a structural and quantitative readout 530:566 a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 530:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 4 59 from development 821:831 arg1 animals 854:860 the tested animals 843:860 the tested animals 843:860 Histopathological studies were carried out to ensure the development of HCC in the tested animals. 26034352 10 60 theme high 1859:1862 arg1 glycans 1872:1878 high mannose glycans 1859:1878 high mannose glycans 1859:1878 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 10 61 theme liver 1918:1922 arg1 tissues 1924:1930 HCC liver tissues 1914:1930 HCC liver tissues 1914:1930 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 3 62 theme glycans 590:596 arg1 readout 560:566 a structural and quantitative readout 530:566 a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 530:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 8 63 theme HCC 1411:1413 arg1 rats 1415:1418 HCC rats 1411:1418 HCC rats 1411:1418 Importantly, HCC rats showed an increase (P < 0.05) in both tumor-associated carbohydrates and in branched glycans. 26034352 10 64 theme reported 1759:1766 arg1 changes 1783:1789 The reported HCC-associated changes 1755:1789 The reported HCC-associated changes in glycan flow and subcellular localization 1755:1833 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 65 theme activities 1664:1673 arg1 importance 1643:1652 the importance 1639:1652 the importance of enzyme activities involved in glycan synthesis at different subcellular localizations 1639:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 7 66 theme high-mannose 1324:1335 arg1 structures 1337:1346 high-mannose structures 1324:1346 high-mannose structures 1324:1346 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 9 67 from changes 1518:1524 arg1 glycans 1529:1535 glycans 1529:1535 glycans 1529:1535 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 10 68 theme glycan 1794:1799 arg1 flow 1801:1804 glycan flow 1794:1804 glycan flow 1794:1804 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 8 69 theme P 1440:1440 arg1 < 1442:1442 P < 0.05 1440:1447 P < 0.05 1440:1447 Importantly, HCC rats showed an increase (P < 0.05) in both tumor-associated carbohydrates and in branched glycans. 26034352 8 69 theme P 1440:1440 arg1 increase 1430:1437 an increase 1427:1437 an increase (P < 0.05) 1427:1448 Importantly, HCC rats showed an increase (P < 0.05) in both tumor-associated carbohydrates and in branched glycans. 26034352 6 70 theme P 1212:1212 arg1 < 1214:1214 P < 0.01 1212:1219 P < 0.01 1212:1219 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 10 71 from tissues 1924:1930 arg1 common 1904:1909 common 1904:1909 common 1904:1909 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 9 72 theme different 1707:1715 arg1 localizations 1729:1741 different subcellular localizations 1707:1741 different subcellular localizations 1707:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 2 73 dep METHODS 301:307 arg1 tissues 315:321 Liver tissues 309:321 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; 301:389 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 7 74 dep ± 1289:1289 arg1 < 1313:1313 P < 0.05 1311:1318 P < 0.05 1311:1318 An increase (66.5 ± 1.05 vs 62.7 ± 1.1, P < 0.05) in high-mannose structures in HCC rats was observed compared to normal rats. 26034352 0 75 from carcinoma 71:79 arg1 rats 84:87 rats 84:87 rats 84:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 3 76 dep unique 606:611 arg1 automated 614:622 automated 614:622 automated 614:622 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 0 77 dep analysis 101:108 arg1 Insights 0:7 Insights 0:7 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats 0:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 6 78 theme P 1139:1139 arg1 < 1141:1141 P < 0.05 1139:1146 P < 0.05 1139:1146 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 8 79 theme tumor-associated 1458:1473 arg1 carbohydrates 1475:1487 both tumor-associated carbohydrates 1453:1487 both tumor-associated carbohydrates 1453:1487 Importantly, HCC rats showed an increase (P < 0.05) in both tumor-associated carbohydrates and in branched glycans. 26034352 9 80 theme glycan 1558:1563 arg1 changes 1570:1576 glycan flow changes 1558:1576 glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations 1558:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 1 81 with association 213:223 arg1 HCC 284:286 HCC 284:286 HCC 284:286 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 1 81 with association 213:223 arg1 carcinoma 273:281 diethyl nitrosamine-induced hepatocellular carcinoma 230:281 diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents 230:298 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 8 82 theme branched 1496:1503 arg1 glycans 1505:1511 branched glycans 1496:1511 branched glycans 1496:1511 Importantly, HCC rats showed an increase (P < 0.05) in both tumor-associated carbohydrates and in branched glycans. 26034352 0 83 theme chemically-induced 37:54 arg1 carcinoma 71:79 chemically-induced hepatocellular carcinoma 37:79 chemically-induced hepatocellular carcinoma in rats 37:87 Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis. 26034352 2 84 theme GlycanMap 441:449 arg1 analyses 451:458 histological and GlycanMap analyses 424:458 analyses 451:458 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 3 85 theme 96-well 624:630 arg1 technology 638:647 a unique, automated 96-well assay technology 604:647 a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 604:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 4 86 theme tested 847:852 arg1 animals 854:860 the tested animals 843:860 the tested animals 843:860 Histopathological studies were carried out to ensure the development of HCC in the tested animals. 26034352 2 87 theme rats 385:388 arg1 tissues 315:321 Liver tissues 309:321 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; 301:389 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 9 88 from localizations 1729:1741 arg1 synthesis 1694:1702 glycan synthesis 1687:1702 glycan synthesis at different subcellular localizations 1687:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 5 89 theme normal 1088:1093 arg1 tissues 1101:1107 normal liver tissues 1088:1107 normal liver tissues 1088:1107 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 2 90 theme histological 424:435 arg1 analyses 451:458 histological and GlycanMap analyses 424:458 analyses 451:458 METHODS Liver tissues of (1) normal (non-tumor-bearing) rats; and (2) tumor-bearing rats; were collected and were used for histological and GlycanMap analyses. 26034352 10 91 theme mannose 1864:1870 arg1 glycans 1872:1878 high mannose glycans 1859:1878 high mannose glycans 1859:1878 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 5 92 dep glycans 906:912 arg1 Gal₂Man₃GlcNac₄Neu₃OAc₃ 970:992 Gal₂Man₃GlcNac₄Neu₃OAc₃ 970:992 Gal₂Man₃GlcNac₄Neu₃OAc₃ 970:992 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 5 92 dep glycans 906:912 arg1 Man₄GlcNac₂ 957:967 Man₄GlcNac₂ 957:967 Man₄GlcNac₂ 957:967 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 5 92 dep glycans 906:912 arg1 glycans 906:912 5 glycans 904:912 5 glycans 904:912 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 5 92 dep glycans 906:912 arg1 Fuc₁ 1011:1014 Fuc₁ 1011:1014 Fuc₁ 1011:1014 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 5 92 dep glycans 906:912 arg1 Gal₂Man₃GlcNac₄Fuc₁Neu₁ 932:954 Gal₂Man₃GlcNac₄Fuc₁Neu₁ 932:954 Gal₂Man₃GlcNac₄Fuc₁Neu₁ 932:954 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 3 93 theme GlycanMap 470:478 arg1 assay 510:514 a high-throughput assay 492:514 a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 492:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 3 93 theme GlycanMap 470:478 arg1 analysis 480:487 GlycanMap analysis 470:487 GlycanMap analysis 470:487 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 5 94 theme rat 1053:1055 arg1 tissues 1061:1067 rat HCC tissues 1053:1067 rat HCC tissues 1053:1067 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26034352 1 95 theme nitrosamine-induced 238:256 arg1 HCC 284:286 HCC 284:286 HCC 284:286 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 1 95 theme nitrosamine-induced 238:256 arg1 carcinoma 273:281 diethyl nitrosamine-induced hepatocellular carcinoma 230:281 diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents 230:298 AIM To evaluate the qualitative and quantitative changes in N-linked glycosylation, which occurred in association with diethyl nitrosamine-induced hepatocellular carcinoma (HCC) in rodents. 26034352 9 96 theme flow 1565:1568 arg1 changes 1570:1576 glycan flow changes 1558:1576 glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations 1558:1741 The changes in glycans correlated well with glycan flow changes reported in the glycan biosynthetic pathway, which indicates the importance of enzyme activities involved in glycan synthesis at different subcellular localizations. 26034352 10 97 theme Lewis 1890:1894 arg1 glycans 1896:1902 siayl Lewis glycans 1884:1902 siayl Lewis glycans 1884:1902 CONCLUSION The reported HCC-associated changes in glycan flow and subcellular localization explain the increase in high mannose glycans and siayl Lewis glycans common in HCC liver tissues. 26034352 6 98 dep decreased 1173:1181 arg1 ± 1189:1189 5.89 ± 0.45 vs 3.54 ± 0.21 1184:1209 5.89 ± 0.45 vs 3.54 ± 0.21 1184:1209 Four glycans were increased (P < 0.05) and Glc₁Man₉GlcNAc₂ was decreased (5.89 ± 0.45 vs 3.54 ± 0.21, P < 0.01) in HCC tissues compared to normal liver tissues. 26034352 3 99 theme high-throughput 494:508 arg1 assay 510:514 a high-throughput assay 492:514 a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics 492:761 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 3 99 theme high-throughput 494:508 arg1 analysis 480:487 GlycanMap analysis 470:487 GlycanMap analysis 470:487 Briefly, GlycanMap analysis is a high-throughput assay that provides a structural and quantitative readout of protein-associated glycans using a unique, automated 96-well assay technology coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and custom bioinformatics. 26034352 5 100 from changes 1042:1048 arg1 tissues 1061:1067 rat HCC tissues 1053:1067 rat HCC tissues 1053:1067 RESULTS The N-glycomic analysis revealed 5 glycans; Glc₁Man₉GlcNAc₂, Gal₂Man₃GlcNac₄Fuc₁Neu₁, Man₄GlcNac₂, Gal₂Man₃GlcNac₄Neu₃OAc₃, and Man₃GlcNac₅ Fuc₁, which showed significant changes in rat HCC tissues when compared with normal liver tissues. 26098314 0 0 theme innovative 83:92 arg1 strategy 94:101 an innovative strategy 80:101 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 0:126 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 4 1 theme immature 636:643 arg1 O-glycans 655:663 immature truncated O-glycans 636:663 immature truncated O-glycans 636:663 Neoplastic transformation frequently shows the expression of immature truncated O-glycans. 26098314 1 2 theme abundant 161:168 arg1 forms 170:174 the most abundant forms 152:174 the most abundant forms of protein posttranslational modification 152:216 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 5 3 theme oncogenic 729:737 arg1 properties 739:748 oncogenic properties 729:748 oncogenic properties 729:748 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 3 4 theme glycan 428:433 arg1 structure 435:443 glycan structure 428:443 glycan structure 428:443 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 3 5 theme structure 435:443 arg1 understanding 383:395 The understanding 379:395 The understanding of protein attachment site and glycan structure 379:443 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 3 6 theme protein 400:406 arg1 site 419:422 protein attachment site 400:422 protein attachment site 400:422 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 7 7 theme cancer 1246:1251 arg1 field 1253:1257 the cancer field 1242:1257 the cancer field 1242:1257 This article will also discuss possible applications of these biomarkers in the cancer field. 26098314 4 8 theme truncated 645:653 arg1 O-glycans 655:663 immature truncated O-glycans 636:663 immature truncated O-glycans 636:663 Neoplastic transformation frequently shows the expression of immature truncated O-glycans. 26098314 7 9 theme biomarkers 1228:1237 arg1 applications 1206:1217 possible applications 1197:1217 possible applications of these biomarkers in the cancer field 1197:1257 This article will also discuss possible applications of these biomarkers in the cancer field. 26098314 5 10 theme expressed 683:691 arg1 O-glycans 693:701 These aberrantly expressed O-glycans 666:701 These aberrantly expressed O-glycans 666:701 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 3 11 theme utmost 455:460 arg1 importance 462:471 the utmost importance 451:471 the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions 451:572 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 1 12 theme forms 170:174 arg1 forms 170:174 the most abundant forms 152:174 the most abundant forms of protein posttranslational modification 152:216 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 1 12 theme forms 170:174 arg1 one 145:147 one 145:147 one 145:147 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 0 13 theme biomarker 107:115 arg1 discovery 117:125 biomarker discovery 107:125 biomarker discovery 107:125 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 6 14 theme homogenous 965:974 arg1 O-glycans 976:984 simplified homogenous O-glycans 954:984 simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites 954:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 15 theme cell 931:934 arg1 models 936:941 genetically engineered cancer cell models 901:941 genetically engineered cancer cell models 901:941 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 16 theme different 288:296 arg1 types 298:302 types 298:302 types 298:302 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 7 17 from applications 1206:1217 arg1 field 1253:1257 the cancer field 1242:1257 the cancer field 1242:1257 This article will also discuss possible applications of these biomarkers in the cancer field. 26098314 0 18 theme cell 16:19 arg1 models 21:26 Glycoengineered cell models 0:26 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 0:126 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 6 19 theme simplified 954:963 arg1 O-glycans 976:984 simplified homogenous O-glycans 954:984 simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites 954:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 20 theme physiologic 340:350 arg1 conditions 367:376 several physiologic and pathologic conditions 332:376 several physiologic and pathologic conditions 332:376 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 1 21 theme protein 179:185 arg1 modification 205:216 protein posttranslational modification 179:216 protein posttranslational modification 179:216 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 0 22 theme Glycoengineered 0:14 arg1 models 21:26 Glycoengineered cell models 0:26 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 0:126 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 6 23 theme cells 1026:1030 arg1 identification 1099:1112 the identification 1095:1112 the identification of potential cancer-specific O-glycosylation sites 1095:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 23 theme cells 1026:1030 arg1 characterization 999:1014 the characterization 995:1014 the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods 995:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 24 theme several 332:338 arg1 conditions 367:376 several physiologic and pathologic conditions 332:376 several physiologic and pathologic conditions 332:376 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 1 25 theme posttranslational 187:203 arg1 modification 205:216 protein posttranslational modification 179:216 protein posttranslational modification 179:216 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 3 26 theme attachment 408:417 arg1 site 419:422 protein attachment site 400:422 protein attachment site 400:422 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 6 27 theme spectrometry 1070:1081 arg1 methods 1083:1089 advanced mass spectrometry methods 1056:1089 advanced mass spectrometry methods 1056:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 7 28 theme possible 1197:1204 arg1 applications 1206:1217 possible applications 1197:1217 possible applications of these biomarkers in the cancer field 1197:1257 This article will also discuss possible applications of these biomarkers in the cancer field. 26098314 1 29 theme modification 205:216 arg1 forms 170:174 the most abundant forms 152:174 the most abundant forms of protein posttranslational modification 152:216 Glycosylation is one of the most abundant forms of protein posttranslational modification. 26098314 3 30 theme site 419:422 arg1 understanding 383:395 The understanding 379:395 The understanding of protein attachment site and glycan structure 379:443 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 6 31 theme mass 1065:1068 arg1 methods 1083:1089 advanced mass spectrometry methods 1056:1089 advanced mass spectrometry methods 1056:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 5 32 theme premalignant 773:784 arg1 meaning 795:801 meaning 795:801 meaning that they are an important source of biomarkers 795:849 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 5 32 theme premalignant 773:784 arg1 lesions 786:792 premalignant lesions 773:792 premalignant lesions 773:792 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 6 33 theme cancer 924:929 arg1 models 936:941 genetically engineered cancer cell models 901:941 genetically engineered cancer cell models 901:941 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 3 34 theme role 502:505 arg1 clarification 481:493 the clarification 477:493 the clarification of the role glycosylation plays in normal cells and in pathological conditions 477:572 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 5 35 theme biomarkers 840:849 arg1 they 808:811 they 808:811 they 808:811 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 5 35 theme biomarkers 840:849 arg1 source 830:835 an important source 817:835 an important source of biomarkers 817:849 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 6 36 theme advanced 1056:1063 arg1 methods 1083:1089 advanced mass spectrometry methods 1056:1089 advanced mass spectrometry methods 1056:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 37 dep cells 1026:1030 arg1 O-glycoproteomes 1032:1047 O-glycoproteomes 1032:1047 O-glycoproteomes 1032:1047 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 38 theme models 936:941 arg1 application 886:896 the recent application 875:896 the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites 875:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 4 39 theme Neoplastic 575:584 arg1 transformation 586:599 Neoplastic transformation 575:599 Neoplastic transformation 575:599 Neoplastic transformation frequently shows the expression of immature truncated O-glycans. 26098314 0 40 theme cancer 56:61 arg1 O-glycoproteome 63:77 cancer O-glycoproteome 56:77 cancer O-glycoproteome 56:77 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 5 41 theme important 820:828 arg1 they 808:811 they 808:811 they 808:811 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 5 41 theme important 820:828 arg1 source 830:835 an important source 817:835 an important source of biomarkers 817:849 These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. 26098314 2 42 theme major 240:244 arg1 type 246:249 a major type 238:249 a major type 238:249 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 2 42 theme major 240:244 arg1 O-glycosylation 219:233 O-glycosylation 219:233 O-glycosylation 219:233 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 4 43 theme O-glycans 655:663 arg1 expression 622:631 the expression 618:631 the expression of immature truncated O-glycans 618:663 Neoplastic transformation frequently shows the expression of immature truncated O-glycans. 26098314 3 44 theme normal 530:535 arg1 cells 537:541 normal cells 530:541 normal cells 530:541 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 3 45 dep role 502:505 arg1 plays 521:525 plays 521:525 plays in normal cells and in pathological conditions 521:572 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 6 46 theme engineered 913:922 arg1 models 936:941 genetically engineered cancer cell models 901:941 genetically engineered cancer cell models 901:941 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 47 theme recent 879:884 arg1 application 886:896 the recent application 875:896 the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites 875:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 48 theme pathologic 356:365 arg1 conditions 367:376 several physiologic and pathologic conditions 332:376 several physiologic and pathologic conditions 332:376 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 6 49 theme sites 1159:1163 arg1 identification 1099:1112 the identification 1095:1112 the identification of potential cancer-specific O-glycosylation sites 1095:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 49 theme sites 1159:1163 arg1 characterization 999:1014 the characterization 995:1014 the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods 995:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 0 50 dep models 21:26 arg1 strategy 94:101 an innovative strategy 80:101 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 0:126 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 6 51 gly O-glycosylation 1143:1157 arg2 sites 1159:1163 potential cancer-specific O-glycosylation sites 1117:1163 potential cancer-specific O-glycosylation sites 1117:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 6 52 theme O-glycosylation 1143:1157 arg1 sites 1159:1163 potential cancer-specific O-glycosylation sites 1117:1163 potential cancer-specific O-glycosylation sites 1117:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 53 theme glycosylation 262:274 arg1 type 246:249 a major type 238:249 a major type 238:249 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 2 53 theme glycosylation 262:274 arg1 O-glycosylation 219:233 O-glycosylation 219:233 O-glycosylation 219:233 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 6 54 theme cancer-specific 1127:1141 arg1 sites 1159:1163 potential cancer-specific O-glycosylation sites 1117:1163 potential cancer-specific O-glycosylation sites 1117:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 0 55 theme O-glycoproteome 63:77 arg1 characterization 36:51 the characterization 32:51 the characterization of cancer O-glycoproteome 32:77 Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery. 26098314 6 56 theme cancer 1019:1024 arg1 cells 1026:1030 cancer cells 1019:1030 cancer cells O-glycoproteomes, using advanced mass spectrometry methods 1019:1089 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 26098314 2 57 theme protein 254:260 arg1 glycosylation 262:274 protein glycosylation 254:274 protein glycosylation 254:274 O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. 26098314 3 58 theme pathological 550:561 arg1 conditions 563:572 pathological conditions 550:572 pathological conditions 550:572 The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. 26098314 6 59 theme potential 1117:1125 arg1 sites 1159:1163 potential cancer-specific O-glycosylation sites 1117:1163 potential cancer-specific O-glycosylation sites 1117:1163 This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. 24831691 8 0 theme ancient 1353:1359 arg1 glycosylation 1361:1373 ancient glycosylation 1353:1373 ancient glycosylation 1353:1373 This discovery clearly shows that glycans persist for thousands of years, and these samples provide a vital insight into ancient glycosylation, offering us a window into the distant past. 24831691 4 1 from evolution 511:519 arg1 systems 597:603 any other biological systems 576:603 any other biological systems 576:603 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 4 1 from evolution 511:519 arg1 humans 555:560 humans 555:560 humans 555:560 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 5 2 theme Scythian 824:831 arg1 " 840:840 the 2,400 year old "Scythian warrior" 804:840 the 2,400 year old "Scythian warrior" 804:840 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 0 3 theme Iceman 78:83 arg1 Princess 95:102 Tyrolean Iceman, Scythian Princess 69:102 Princess 95:102 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 0 3 theme Iceman 78:83 arg1 Scythian 86:93 Scythian 86:93 Scythian 86:93 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 5 4 gly glycosylation 637:649 arg1 study 614:618 this study 609:618 this study 609:618 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 4 gly glycosylation 637:649 arg1 samples 661:667 tissue samples 654:667 tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria 654:953 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 5 theme warrior 833:839 arg1 " 840:840 the 2,400 year old "Scythian warrior" 804:840 the 2,400 year old "Scythian warrior" 804:840 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 0 6 theme Princess 95:102 arg1 analysis 57:64 N-glycan analysis 48:64 N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior 48:114 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 7 7 from sample 1129:1134 arg1 significant 1164:1174 significant 1164:1174 significant 1164:1174 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 6 8 theme mummies 1061:1067 arg1 age 1022:1024 the age 1018:1024 the age 1018:1024 The number of N-glycans that were identified varied both with the age and the preservation status of the mummies. 24831691 6 8 theme mummies 1061:1067 arg1 status 1047:1052 the preservation status 1030:1052 the preservation status of the mummies 1030:1067 The number of N-glycans that were identified varied both with the age and the preservation status of the mummies. 24831691 8 9 theme years 1299:1303 arg1 thousands 1286:1294 thousands 1286:1294 thousands of years 1286:1303 This discovery clearly shows that glycans persist for thousands of years, and these samples provide a vital insight into ancient glycosylation, offering us a window into the distant past. 24831691 4 10 theme other 580:584 arg1 systems 597:603 any other biological systems 576:603 any other biological systems 576:603 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 6 11 theme preservation 1034:1045 arg1 status 1047:1052 the preservation status 1030:1052 the preservation status of the mummies 1030:1067 The number of N-glycans that were identified varied both with the age and the preservation status of the mummies. 24831691 5 12 located found 776:780 arg2 " 773:773 Oetzi" 768:773 Oetzi" 768:773 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 12 located found 776:780 arg1 Alps 798:801 the Tyrolean Alps 785:801 the Tyrolean Alps 785:801 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 7 13 theme contemporary 1116:1127 arg1 sample 1129:1134 the contemporary sample 1112:1134 the contemporary sample 1112:1134 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 2 14 from analysis 225:232 arg1 samples 270:276 biological samples 259:276 biological samples 259:276 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 5 15 theme Scythian 847:854 arg1 " 864:864 "Scythian Princess" 846:864 "Scythian Princess" 846:864 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 0 16 theme Warrior 108:114 arg1 analysis 57:64 N-glycan analysis 48:64 N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior 48:114 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 5 17 dep Iceman 754:759 arg1 called 761:766 called 761:766 called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy 761:928 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 2 18 theme strict 281:286 arg1 regime 288:293 a strict regime 279:293 a strict regime 279:293 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 1 19 theme new 173:175 arg1 insights 177:184 new insights 173:184 new insights into many biological processes 173:215 An improved understanding of glycosylation will provide new insights into many biological processes. 24831691 5 20 dep samples 661:667 arg1 Iceman 754:759 - the 5,300 year old "Iceman 732:759 Iceman 754:759 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 21 theme old 749:751 arg1 Iceman 754:759 - the 5,300 year old "Iceman 732:759 Iceman 754:759 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 6 22 theme N-glycans 970:978 arg1 number 960:965 The number 956:965 The number of N-glycans that were identified 956:999 The number of N-glycans that were identified varied both with the age and the preservation status of the mummies. 24831691 5 23 located found 867:871 arg2 " 840:840 the 2,400 year old "Scythian warrior" 804:840 the 2,400 year old "Scythian warrior" 804:840 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 23 located found 867:871 arg1 Mountains 886:894 Mountains 886:894 Mountains 886:894 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 24 theme Oetzi 768:772 arg1 " 773:773 Oetzi" 768:773 Oetzi" 768:773 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 25 theme Princess 856:863 arg1 " 864:864 "Scythian Princess" 846:864 "Scythian Princess" 846:864 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 0 26 theme Glycosylated 0:11 arg1 proteins 13:20 Glycosylated proteins 0:20 Glycosylated proteins 0:20 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 1 27 theme many 191:194 arg1 processes 207:215 many biological processes 191:215 many biological processes 191:215 An improved understanding of glycosylation will provide new insights into many biological processes. 24831691 3 28 theme glycans 366:372 arg1 fate 358:361 the fate 354:361 the fate of glycans that have been exposed to environmental conditions over millennia 354:438 However, the fate of glycans that have been exposed to environmental conditions over millennia has not yet been investigated. 24831691 7 29 theme ancient 1208:1214 arg1 samples 1223:1229 the ancient tissue samples 1204:1229 the ancient tissue samples 1204:1229 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 1 30 theme biological 196:205 arg1 processes 207:215 many biological processes 191:215 many biological processes 191:215 An improved understanding of glycosylation will provide new insights into many biological processes. 24831691 5 31 attach derived 669:675 arg1 mummies 687:693 four mummies 682:693 four mummies which have been naturally preserved 682:729 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 31 attach derived 669:675 arg2 samples 661:667 tissue samples 654:667 tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria 654:953 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 3 32 theme environmental 400:412 arg1 conditions 414:423 environmental conditions 400:423 environmental conditions 400:423 However, the fate of glycans that have been exposed to environmental conditions over millennia has not yet been investigated. 24831691 2 33 theme oligosaccharides 237:252 arg1 analysis 225:232 the analysis 221:232 the analysis of oligosaccharides from biological samples 221:276 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 2 34 from samples 270:276 arg1 oligosaccharides 237:252 oligosaccharides 237:252 oligosaccharides from biological samples 237:276 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 2 34 from samples 270:276 arg1 analysis 225:232 the analysis 221:232 the analysis of oligosaccharides from biological samples 221:276 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 5 35 theme Tyrolean 789:796 arg1 Alps 798:801 the Tyrolean Alps 785:801 the Tyrolean Alps 785:801 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 36 theme old 910:912 arg1 mummy 924:928 a 4 year old apartment mummy 901:928 a 4 year old apartment mummy 901:928 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 7 37 theme More 1070:1073 arg1 structures 1082:1091 More glycan structures 1070:1091 More glycan structures 1070:1091 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 7 38 theme glycan 1075:1080 arg1 structures 1082:1091 More glycan structures 1070:1091 More glycan structures 1070:1091 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 5 39 theme tissue 654:659 arg1 samples 661:667 tissue samples 654:667 tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria 654:953 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 5 40 theme apartment 914:922 arg1 mummy 924:928 a 4 year old apartment mummy 901:928 a 4 year old apartment mummy 901:928 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 7 41 from significant 1164:1174 arg1 sample 1129:1134 the contemporary sample 1112:1134 the contemporary sample 1112:1134 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 0 42 theme N-glycan 48:55 arg1 analysis 57:64 N-glycan analysis 48:64 N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior 48:114 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 4 43 theme machinery 542:550 arg1 evolution 511:519 the evolution 507:519 the evolution of the glycosylation machinery in humans as well as in any other biological systems 507:603 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 5 44 theme samples 661:667 arg1 glycosylation 637:649 the glycosylation 633:649 the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria 633:953 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 8 45 theme vital 1334:1338 arg1 insight 1340:1346 a vital insight 1332:1346 a vital insight into ancient glycosylation 1332:1373 This discovery clearly shows that glycans persist for thousands of years, and these samples provide a vital insight into ancient glycosylation, offering us a window into the distant past. 24831691 5 46 dep called 761:766 arg1 found 931:935 found 931:935 found in Vienna/Austria 931:953 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 4 47 theme glycosylation 528:540 arg1 machinery 542:550 the glycosylation machinery 524:550 the glycosylation machinery 524:550 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 1 48 theme improved 120:127 arg1 understanding 129:141 An improved understanding 117:141 An improved understanding of glycosylation 117:158 An improved understanding of glycosylation will provide new insights into many biological processes. 24831691 0 49 dep preserved 22:30 arg1 analysis 57:64 N-glycan analysis 48:64 N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior 48:114 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 2 50 theme sample 327:332 arg1 integrity 334:342 sample integrity 327:342 sample integrity 327:342 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 4 51 theme biological 586:595 arg1 systems 597:603 any other biological systems 576:603 any other biological systems 576:603 This is also true for understanding the evolution of the glycosylation machinery in humans as well as in any other biological systems. 24831691 0 52 theme Tyrolean 69:76 arg1 Princess 95:102 Tyrolean Iceman, Scythian Princess 69:102 Princess 95:102 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 0 52 theme Tyrolean 69:76 arg1 Scythian 86:93 Scythian 86:93 Scythian 86:93 Glycosylated proteins preserved over millennia: N-glycan analysis of Tyrolean Iceman, Scythian Princess and Warrior. 24831691 2 53 theme biological 259:268 arg1 samples 270:276 biological samples 259:276 biological samples 259:276 In the analysis of oligosaccharides from biological samples, a strict regime is typically followed to ensure sample integrity. 24831691 5 54 theme old 819:821 arg1 " 840:840 the 2,400 year old "Scythian warrior" 804:840 the 2,400 year old "Scythian warrior" 804:840 In this study, we examined the glycosylation of tissue samples derived from four mummies which have been naturally preserved: - the 5,300 year old "Iceman called Oetzi", found in the Tyrolean Alps; the 2,400 year old "Scythian warrior" and "Scythian Princess", found in the Altai Mountains; and a 4 year old apartment mummy, found in Vienna/Austria. 24831691 7 55 theme tissue 1216:1221 arg1 samples 1223:1229 the ancient tissue samples 1204:1229 the ancient tissue samples 1204:1229 More glycan structures were discovered in the contemporary sample, as expected, however it is significant that glycan still exists in the ancient tissue samples. 24831691 1 56 theme glycosylation 146:158 arg1 understanding 129:141 An improved understanding 117:141 An improved understanding of glycosylation 117:158 An improved understanding of glycosylation will provide new insights into many biological processes. 24831691 8 57 dep us 1385:1386 arg1 window 1390:1395 a window 1388:1395 us a window into the distant past 1385:1417 This discovery clearly shows that glycans persist for thousands of years, and these samples provide a vital insight into ancient glycosylation, offering us a window into the distant past. 25303961 6 0 theme O-linked 834:841 arg1 glycans 843:849 the 8 O-linked glycans 828:849 the 8 O-linked glycans detected 828:858 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 9 1 theme DR 1473:1474 arg1 development 1445:1455 the development 1441:1455 the development of diabetes and DR 1441:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 1 2 from important 162:170 arg1 role 179:182 the role 175:182 the role of immunity of the ocular surface 175:216 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 1 3 theme glycosylation 132:144 arg1 state 118:122 the state 114:122 the state of tear glycosylation 114:144 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 1 3 theme glycosylation 132:144 arg1 important 162:170 important 162:170 important 162:170 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 0 4 theme diabetic 71:78 arg1 retinopathy 80:90 diabetic retinopathy 71:90 diabetic retinopathy 71:90 Glycomic characterization of basal tears and changes with diabetes and diabetic retinopathy. 25303961 8 5 link N-linked 1144:1151 arg1 glycans 1153:1159 five low abundance N-linked glycans 1125:1159 five low abundance N-linked glycans 1125:1159 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 3 6 with patients 452:459 arg1 diabetes 513:520 diabetes 513:520 diabetes 513:520 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 6 with patients 452:459 arg1 retinopathy 526:536 retinopathy 526:536 retinopathy 526:536 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 6 with patients 452:459 arg1 diabetes 466:473 diabetes 466:473 diabetes without retinopathy 466:493 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 8 7 theme mellitus 1220:1227 arg1 onset 1202:1206 the onset 1198:1206 the onset of diabetes mellitus and diabetic retinopathy (DR) 1198:1257 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 6 8 theme core 888:891 arg1 type 895:898 core 2 type 888:898 core 2 type 888:898 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 5 9 theme glycans 683:689 arg1 complex 707:713 complex 707:713 complex 707:713 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 2 10 theme biomarkers 263:272 arg1 source 240:245 a valuable source 229:245 a valuable source of non-invasive biomarkers for disease 229:284 Tears are a valuable source of non-invasive biomarkers for disease and there are continued efforts to characterize their components thoroughly. 25303961 2 10 theme biomarkers 263:272 arg1 Tears 219:223 Tears 219:223 Tears 219:223 Tears are a valuable source of non-invasive biomarkers for disease and there are continued efforts to characterize their components thoroughly. 25303961 4 11 theme relative 615:622 arg1 abundance 624:632 the relative abundance 611:632 the relative abundance of each structure 611:650 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 7 12 theme glycan 969:974 arg1 structures 976:985 these glycan structures 963:985 these glycan structures 963:985 Additionally, these glycan structures were profiled across the three diabetic disease groups. 25303961 2 13 theme non-invasive 250:261 arg1 biomarkers 263:272 non-invasive biomarkers 250:272 non-invasive biomarkers for disease 250:284 Tears are a valuable source of non-invasive biomarkers for disease and there are continued efforts to characterize their components thoroughly. 25303961 4 14 theme detailed 543:550 arg1 N- 552:553 The detailed N- 539:553 The detailed N- 539:553 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 8 15 theme N-linked 1144:1151 arg1 glycans 1153:1159 five low abundance N-linked glycans 1125:1159 five low abundance N-linked glycans 1125:1159 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 9 16 theme types 1309:1313 arg1 conservation 1286:1297 the conservation 1282:1297 the conservation of glycan types on basal tear proteins between individuals 1282:1356 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 9 17 from proteins 1329:1336 arg1 conservation 1286:1297 the conservation 1282:1297 the conservation of glycan types on basal tear proteins between individuals 1282:1356 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 8 18 theme higher 1054:1059 arg1 structures 1070:1079 the higher abundant structures 1050:1079 the higher abundant structures 1050:1079 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 3 19 theme tears 402:406 arg1 volume 386:391 a small volume 378:391 a small volume of basal tears (5 μL) 378:413 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 9 20 with tears 1430:1434 arg1 development 1445:1455 the development 1441:1455 the development of diabetes and DR 1441:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 8 21 link O-linked 1167:1174 arg1 glycan 1176:1181 1 O-linked glycan 1165:1181 1 O-linked glycan 1165:1181 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 9 22 theme glycan 1393:1398 arg1 expression 1400:1409 glycan expression 1393:1409 glycan expression on the proteins in tears with the development of diabetes and DR 1393:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 5 23 theme bisecting 737:745 arg1 N-acetylglucosamine 747:765 a bisecting N-acetylglucosamine 735:765 a bisecting N-acetylglucosamine 735:765 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 23 theme bisecting 737:745 arg1 % 770:770 65% 768:770 65% containing a core fucose 768:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 8 24 theme diabetes 1211:1218 arg1 mellitus 1220:1227 diabetes mellitus 1211:1227 diabetes mellitus 1211:1227 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 0 25 theme Glycomic 0:7 arg1 characterization 9:24 Glycomic characterization 0:24 Glycomic characterization of basal tears 0:39 Glycomic characterization of basal tears and changes with diabetes and diabetic retinopathy. 25303961 3 26 with controls 442:449 arg1 diabetes 513:520 diabetes 513:520 diabetes 513:520 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 26 with controls 442:449 arg1 retinopathy 526:536 retinopathy 526:536 retinopathy 526:536 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 26 with controls 442:449 arg1 diabetes 466:473 diabetes 466:473 diabetes without retinopathy 466:493 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 5 27 contain containing 772:781 arg1 N-acetylglucosamine 747:765 a bisecting N-acetylglucosamine 735:765 a bisecting N-acetylglucosamine 735:765 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 27 contain containing 772:781 arg1 % 770:770 65% 768:770 65% containing a core fucose 768:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 27 contain containing 772:781 arg2 fucose 790:795 a core fucose 783:795 a core fucose 783:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 6 28 dep sialylated 931:940 arg1 % 945:945 90% 943:945 90% 943:945 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 3 29 theme basal 396:400 arg1 tears 402:406 basal tears 396:406 basal tears (5 μL) 396:413 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 29 theme basal 396:400 arg1 μL 411:412 5 μL 409:412 5 μL 409:412 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 0 30 theme basal 29:33 arg1 tears 35:39 basal tears 29:39 basal tears 29:39 Glycomic characterization of basal tears and changes with diabetes and diabetic retinopathy. 25303961 8 31 theme abundant 1061:1068 arg1 structures 1070:1079 the higher abundant structures 1050:1079 the higher abundant structures 1050:1079 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 9 32 from changes 1382:1388 arg1 expression 1400:1409 glycan expression 1393:1409 glycan expression on the proteins in tears with the development of diabetes and DR 1393:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 9 32 from changes 1382:1388 arg1 proteins 1418:1425 the proteins 1414:1425 the proteins in tears with the development of diabetes and DR 1414:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 5 33 with complex 707:713 arg1 % 722:722 50% 720:722 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose 720:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 9 34 theme glycan 1302:1307 arg1 types 1309:1313 glycan types 1302:1313 glycan types on basal tear proteins 1302:1336 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 7 35 theme disease 1027:1033 arg1 groups 1035:1040 the three diabetic disease groups 1008:1040 the three diabetic disease groups 1008:1040 Additionally, these glycan structures were profiled across the three diabetic disease groups. 25303961 1 36 theme secreted 98:105 arg1 fluid 107:111 a secreted fluid 96:111 a secreted fluid 96:111 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 6 37 theme them 920:923 arg1 majority 908:915 a majority 906:915 a majority of them 906:923 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 3 38 theme small 380:384 arg1 volume 386:391 a small volume 378:391 a small volume of basal tears (5 μL) 378:413 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 8 39 theme low 1130:1132 arg1 glycans 1153:1159 five low abundance N-linked glycans 1125:1159 five low abundance N-linked glycans 1125:1159 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 8 40 theme retinopathy 1242:1252 arg1 onset 1202:1206 the onset 1198:1206 the onset of diabetes mellitus and diabetic retinopathy (DR) 1198:1257 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 6 41 link O-linked 834:841 arg1 glycans 843:849 the 8 O-linked glycans 828:849 the 8 O-linked glycans detected 828:858 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 1 42 theme immunity 187:194 arg1 role 179:182 the role 175:182 the role of immunity of the ocular surface 175:216 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 0 43 theme tears 35:39 arg1 characterization 9:24 Glycomic characterization 0:24 Glycomic characterization of basal tears 0:39 Glycomic characterization of basal tears and changes with diabetes and diabetic retinopathy. 25303961 5 44 theme N-linked 674:681 arg1 glycans 683:689 the 50 N-linked glycans 667:689 the 50 N-linked glycans found 667:695 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 4 45 link O-linked 559:566 arg1 glycome 581:587 O-linked tear protein glycome 559:587 O-linked tear protein glycome 559:587 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 4 46 theme protein 573:579 arg1 glycome 581:587 O-linked tear protein glycome 559:587 O-linked tear protein glycome 559:587 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 5 47 theme core 785:788 arg1 fucose 790:795 a core fucose 783:795 a core fucose 783:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 48 gly sialylated 813:822 arg1 % 806:806 33% 804:806 33% 804:806 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 9 49 theme tear 1324:1327 arg1 proteins 1329:1336 basal tear proteins 1318:1336 basal tear proteins 1318:1336 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 3 50 theme healthy 434:440 arg1 controls 442:449 healthy controls 434:449 healthy controls 434:449 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 51 with patients 499:506 arg1 diabetes 513:520 diabetes 513:520 diabetes 513:520 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 51 with patients 499:506 arg1 retinopathy 526:536 retinopathy 526:536 retinopathy 526:536 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 3 51 with patients 499:506 arg1 diabetes 466:473 diabetes 466:473 diabetes without retinopathy 466:493 In this study, a small volume of basal tears (5 μL) was collected from healthy controls, patients with diabetes without retinopathy and patients with diabetes and retinopathy. 25303961 9 52 theme small 1376:1380 arg1 changes 1382:1388 only small changes 1371:1388 only small changes in glycan expression on the proteins in tears with the development of diabetes and DR 1371:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 1 53 from role 179:182 arg1 state 118:122 the state 114:122 the state of tear glycosylation 114:144 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 1 53 from role 179:182 arg1 important 162:170 important 162:170 important 162:170 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 8 54 theme abundance 1134:1142 arg1 glycans 1153:1159 five low abundance N-linked glycans 1125:1159 five low abundance N-linked glycans 1125:1159 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 7 55 theme diabetic 1018:1025 arg1 groups 1035:1040 the three diabetic disease groups 1008:1040 the three diabetic disease groups 1008:1040 Additionally, these glycan structures were profiled across the three diabetic disease groups. 25303961 6 56 gly sialylated 931:940 arg1 majority 908:915 a majority 906:915 a majority of them 906:923 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 5 57 contain containing 724:733 arg2 % 770:770 65% 768:770 65% containing a core fucose 768:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 57 contain containing 724:733 arg2 N-acetylglucosamine 747:765 a bisecting N-acetylglucosamine 735:765 a bisecting N-acetylglucosamine 735:765 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 5 57 contain containing 724:733 arg1 % 722:722 50% 720:722 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose 720:795 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 2 58 theme valuable 231:238 arg1 source 240:245 a valuable source 229:245 a valuable source of non-invasive biomarkers for disease 229:284 Tears are a valuable source of non-invasive biomarkers for disease and there are continued efforts to characterize their components thoroughly. 25303961 2 58 theme valuable 231:238 arg1 Tears 219:223 Tears 219:223 Tears 219:223 Tears are a valuable source of non-invasive biomarkers for disease and there are continued efforts to characterize their components thoroughly. 25303961 8 59 theme diabetic 1233:1240 arg1 DR 1255:1256 DR 1255:1256 DR 1255:1256 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 8 59 theme diabetic 1233:1240 arg1 retinopathy 1242:1252 diabetic retinopathy 1233:1252 diabetic retinopathy (DR) 1233:1257 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 9 60 from proteins 1418:1425 arg1 changes 1382:1388 only small changes 1371:1388 only small changes in glycan expression on the proteins in tears with the development of diabetes and DR 1371:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 9 60 from proteins 1418:1425 arg1 tears 1430:1434 tears 1430:1434 tears with the development of diabetes and DR 1430:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 4 61 theme O-linked 559:566 arg1 glycome 581:587 O-linked tear protein glycome 559:587 O-linked tear protein glycome 559:587 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 9 62 from types 1309:1313 arg1 proteins 1329:1336 basal tear proteins 1318:1336 basal tear proteins 1318:1336 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 8 63 theme O-linked 1167:1174 arg1 glycan 1176:1181 1 O-linked glycan 1165:1181 1 O-linked glycan 1165:1181 Whilst the higher abundant structures did not alter across the three groups, only five low abundance N-linked glycans and 1 O-linked glycan did alter with the onset of diabetes mellitus and diabetic retinopathy (DR). 25303961 9 64 from conservation 1286:1297 arg1 proteins 1329:1336 basal tear proteins 1318:1336 basal tear proteins 1318:1336 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 5 65 link N-linked 674:681 arg1 glycans 683:689 the 50 N-linked glycans 667:689 the 50 N-linked glycans found 667:695 Of the 50 N-linked glycans found, 89% were complex with 50% containing a bisecting N-acetylglucosamine, 65% containing a core fucose whilst 33% were sialylated. 25303961 9 66 from expression 1400:1409 arg1 proteins 1418:1425 the proteins 1414:1425 the proteins in tears with the development of diabetes and DR 1414:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 1 67 theme ocular 203:208 arg1 surface 210:216 the ocular surface 199:216 the ocular surface 199:216 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 9 68 theme basal 1318:1322 arg1 proteins 1329:1336 basal tear proteins 1318:1336 basal tear proteins 1318:1336 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 9 69 theme diabetes 1460:1467 arg1 development 1445:1455 the development 1441:1455 the development of diabetes and DR 1441:1474 These results suggest the conservation of glycan types on basal tear proteins between individuals and point to only small changes in glycan expression on the proteins in tears with the development of diabetes and DR. 25303961 4 70 theme structure 642:650 arg1 abundance 624:632 the relative abundance 611:632 the relative abundance of each structure 611:650 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 25303961 1 71 theme surface 210:216 arg1 immunity 187:194 immunity 187:194 immunity of the ocular surface 187:216 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 6 72 theme glycans 843:849 arg1 cores 871:875 cores 1 and 5 of core 2 type 871:898 cores 1 and 5 of core 2 type 871:898 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 6 73 theme type 895:898 arg1 cores 871:875 cores 1 and 5 of core 2 type 871:898 cores 1 and 5 of core 2 type 871:898 Of the 8 O-linked glycans detected, 3 were of cores 1 and 5 of core 2 type, with a majority of them being sialylated (90%). 25303961 1 74 theme tear 127:130 arg1 glycosylation 132:144 tear glycosylation 127:144 tear glycosylation 127:144 As a secreted fluid, the state of tear glycosylation is particularly important in the role of immunity of the ocular surface. 25303961 4 75 theme tear 568:571 arg1 glycome 581:587 O-linked tear protein glycome 559:587 O-linked tear protein glycome 559:587 The detailed N- and O-linked tear protein glycome was characterized and the relative abundance of each structure determined. 24855066 11 0 theme complex 2118:2124 arg1 association 2126:2136 their complex association 2112:2136 their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer 2112:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 7 1 theme glycan 1264:1269 arg1 features 1271:1278 characteristic glycan features 1249:1278 characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans 1249:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 11 2 from cancer 2215:2220 arg1 programming 2154:2164 epigenetic programming 2143:2164 epigenetic programming of their associated synthetic enzymes in ovarian cancer 2143:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 7 3 theme N 1450:1450 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 3 theme N 1450:1450 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 1 4 theme mortality 261:269 arg1 rate 271:274 the highest mortality rate 249:274 the highest mortality rate among all gynecological cancers 249:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 6 5 theme molecular 1180:1188 arg1 masses 1190:1195 their molecular masses 1174:1195 their molecular masses 1174:1195 Glycan structures were characterized based on their molecular masses and tandem MS fragmentation patterns. 24855066 9 6 from influence 1702:1710 arg1 expression 1721:1730 MGAT3 expression 1715:1730 MGAT3 expression 1715:1730 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 3 7 theme ovarian 552:558 arg1 cancer 560:565 ovarian cancer 552:565 ovarian cancer 552:565 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 3 8 dep development 608:618 arg1 the 604:606 the 604:606 the 604:606 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 7 9 theme " 1475:1475 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 9 theme " 1475:1475 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 9 10 theme epigenetic 1691:1700 arg1 influence 1702:1710 the potential epigenetic influence 1677:1710 the potential epigenetic influence on MGAT3 expression 1677:1730 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 7 11 theme sialylated 1424:1433 arg1 N-glycans 1435:1443 α 2-6 sialylated N-glycans 1418:1443 α 2-6 sialylated N-glycans 1418:1443 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 11 12 theme drug 2289:2292 arg1 targets 2294:2300 novel anti-glycan drug targets 2271:2300 novel anti-glycan drug targets 2271:2300 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 10 13 theme bisecting 1966:1974 arg1 N-glycans 1989:1997 the unique "bisecting GlcNAc" type N-glycans 1954:1997 the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1954:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 2 14 theme Cell 309:312 arg1 glycans 323:329 Cell membrane glycans 309:329 Cell membrane glycans 309:329 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 1 15 theme fifth 188:192 arg1 cancer 174:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 1 15 theme fifth 188:192 arg1 cause 206:210 the fifth most common cause 184:210 the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers 184:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 8 16 dep synthesis 1600:1608 arg1 ST6GAL1 1618:1624 ST6GAL1 1618:1624 ST6GAL1 1618:1624 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 8 16 dep synthesis 1600:1608 arg1 B4GALNT3 1631:1638 B4GALNT3 1631:1638 B4GALNT3 1631:1638 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 8 16 dep synthesis 1600:1608 arg1 MGAT3 1611:1615 MGAT3 1611:1615 MGAT3 1611:1615 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 5 17 attach released 935:942 arg1 glycoproteins 958:970 membrane glycoproteins 949:970 membrane glycoproteins 949:970 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 17 attach released 935:942 arg2 N-glycans 920:928 N-glycans 920:928 N-glycans 920:928 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 10 18 theme unique 1958:1963 arg1 N-glycans 1989:1997 the unique "bisecting GlcNAc" type N-glycans 1954:1997 the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1954:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 1 19 theme common 199:204 arg1 cancer 174:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 1 19 theme common 199:204 arg1 cause 206:210 the fifth most common cause 184:210 the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers 184:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 4 20 theme ovarian 903:909 arg1 cancers 911:917 epithelial ovarian cancers 892:917 epithelial ovarian cancers 892:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 11 21 theme diagnostic 2315:2324 arg1 tools 2326:2330 clinical diagnostic tools 2306:2330 clinical diagnostic tools 2306:2330 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 10 22 theme N-glycans 1989:1997 arg1 synthesis 1941:1949 the synthesis 1937:1949 the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1937:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 0 23 theme DNA 131:133 arg1 status 147:152 DNA methylation status 131:152 DNA methylation status 131:152 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 5 24 theme electrospray 1076:1087 arg1 ESI-MS 1119:1124 ESI-MS 1119:1124 ESI-MS 1119:1124 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 24 theme electrospray 1076:1087 arg1 spectrometry 1105:1116 negative-ion electrospray ionization mass spectrometry 1063:1116 negative-ion electrospray ionization mass spectrometry (ESI-MS) 1063:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 10 25 theme first 1816:1820 arg1 time 1822:1825 the first time 1812:1825 the first time 1812:1825 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 1 26 from cause 206:210 arg1 worldwide 231:239 women worldwide 225:239 women worldwide bearing the highest mortality rate among all gynecological cancers 225:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 11 27 theme N-glycan 2085:2092 arg1 substructures 2094:2106 specific N-glycan substructures 2076:2106 specific N-glycan substructures 2076:2106 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 11 28 used used 2243:2246 arg2 association 2126:2136 their complex association 2112:2136 their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer 2112:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 9 29 theme cell 1748:1751 arg1 lines 1753:1757 the cell lines 1744:1757 the cell lines 1744:1757 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 10 30 theme " 1982:1982 arg1 N-glycans 1989:1997 the unique "bisecting GlcNAc" type N-glycans 1954:1997 the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1954:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 4 31 gly glycosylation 702:714 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 31 gly glycosylation 702:714 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 31 gly glycosylation 702:714 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 31 gly glycosylation 702:714 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 31 gly glycosylation 702:714 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 31 gly glycosylation 702:714 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 32 theme common 869:874 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 32 theme common 869:874 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 7 33 theme type 1382:1385 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 33 theme type 1382:1385 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 33 theme type 1382:1385 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 33 theme type 1382:1385 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 5 34 theme graphitized 1040:1050 arg1 carbon 1052:1057 porous graphitized carbon 1033:1057 porous graphitized carbon 1033:1057 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 8 35 theme N-glycan 1499:1506 arg1 changes 1508:1514 These N-glycan changes 1493:1514 These N-glycan changes 1493:1514 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 7 36 theme N-acetyl-glucosamine 1360:1379 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 36 theme N-acetyl-glucosamine 1360:1379 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 36 theme N-acetyl-glucosamine 1360:1379 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 36 theme N-acetyl-glucosamine 1360:1379 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 37 theme characteristic 1249:1262 arg1 features 1271:1278 characteristic glycan features 1249:1278 characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans 1249:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 5 38 gly glycoproteins 958:970 arg1 glycoproteins 958:970 membrane glycoproteins 949:970 membrane glycoproteins 949:970 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 4 39 theme non-cancerous 719:731 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 39 theme non-cancerous 719:731 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 39 theme non-cancerous 719:731 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 39 theme non-cancerous 719:731 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 39 theme non-cancerous 719:731 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 39 theme non-cancerous 719:731 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 10 40 theme cells 2042:2046 arg1 proteins 2015:2022 the membrane proteins 2002:2022 the membrane proteins of ovarian cancer cells 2002:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 1 41 theme ovarian 166:172 arg1 cancer 174:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 1 41 theme ovarian 166:172 arg1 cause 206:210 the fifth most common cause 184:210 the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers 184:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 9 42 theme DNA 1781:1783 arg1 inhibitor 1797:1805 a DNA methylation inhibitor 1779:1805 a DNA methylation inhibitor 1779:1805 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 9 42 theme DNA 1781:1783 arg1 5-azacytidine 1764:1776 5-azacytidine 1764:1776 5-azacytidine 1764:1776 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 10 43 theme ovarian 2027:2033 arg1 cells 2042:2046 ovarian cancer cells 2027:2046 ovarian cancer cells 2027:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 0 44 theme glycan 85:90 arg1 structures 92:101 glycan structures 85:101 glycan structures 85:101 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 5 45 theme mass 1100:1103 arg1 ESI-MS 1119:1124 ESI-MS 1119:1124 ESI-MS 1119:1124 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 45 theme mass 1100:1103 arg1 spectrometry 1105:1116 negative-ion electrospray ionization mass spectrometry 1063:1116 negative-ion electrospray ionization mass spectrometry (ESI-MS) 1063:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 8 46 theme gene 1543:1546 arg1 levels 1559:1564 gene transcript levels 1543:1564 gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR 1543:1653 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 4 47 theme serous 789:794 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 47 theme serous 789:794 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 47 theme serous 789:794 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 47 theme serous 789:794 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 47 theme serous 789:794 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 47 theme serous 789:794 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 7 48 theme bisecting 1350:1358 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 48 theme bisecting 1350:1358 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 48 theme bisecting 1350:1358 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 48 theme bisecting 1350:1358 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 11 49 theme synthetic 2186:2194 arg1 enzymes 2196:2202 their associated synthetic enzymes 2169:2202 their associated synthetic enzymes in ovarian cancer 2169:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 0 50 theme Specific 0:7 arg1 glycosylation 9:21 Specific glycosylation 0:21 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 0:153 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 4 51 dep epithelial 749:758 arg1 6.3 766:768 HOSE 6.3 761:768 HOSE 6.3 761:768 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 51 dep epithelial 749:758 arg1 17.1 779:782 HOSE 17.1 774:782 HOSE 17.1 774:782 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 6 52 theme MS 1208:1209 arg1 patterns 1225:1232 tandem MS fragmentation patterns 1201:1232 tandem MS fragmentation patterns 1201:1232 Glycan structures were characterized based on their molecular masses and tandem MS fragmentation patterns. 24855066 4 53 theme HOSE 774:777 arg1 17.1 779:782 HOSE 17.1 774:782 HOSE 17.1 774:782 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 8 54 theme specific 1581:1588 arg1 enzymes 1573:1579 the enzymes 1569:1579 the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR 1569:1653 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 0 55 theme proteins 35:42 arg1 glycosylation 9:21 Specific glycosylation 0:21 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 0:153 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 0 56 gly glycosylation 9:21 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 0 56 gly glycosylation 9:21 arg1 proteins 35:42 membrane proteins 26:42 membrane proteins 26:42 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 7 57 theme membrane 1319:1326 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 57 theme membrane 1319:1326 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 0 58 theme cancer 66:71 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 2 59 theme cellular 347:354 arg1 signaling 379:387 cell signaling 374:387 cell signaling 374:387 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 2 59 theme cellular 347:354 arg1 processes 356:364 various cellular processes 339:364 various cellular processes such as cell signaling 339:387 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 0 60 theme epithelial 47:56 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 6 61 theme Glycan 1128:1133 arg1 structures 1135:1144 Glycan structures 1128:1144 Glycan structures 1128:1144 Glycan structures were characterized based on their molecular masses and tandem MS fragmentation patterns. 24855066 6 62 theme tandem 1201:1206 arg1 patterns 1225:1232 tandem MS fragmentation patterns 1201:1232 tandem MS fragmentation patterns 1201:1232 Glycan structures were characterized based on their molecular masses and tandem MS fragmentation patterns. 24855066 4 63 theme cancer 804:809 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 63 theme cancer 804:809 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 63 theme cancer 804:809 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 63 theme cancer 804:809 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 63 theme cancer 804:809 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 63 theme cancer 804:809 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 3 64 theme aberrant 491:498 arg1 regulation 500:509 aberrant regulation 491:509 aberrant regulation of gene expression 491:528 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 1 65 theme cancer 215:220 arg1 cancer 174:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 1 65 theme cancer 215:220 arg1 cause 206:210 the fifth most common cause 184:210 the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers 184:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 5 66 theme PNGase 975:980 arg1 F 982:982 PNGase F 975:982 PNGase F 975:982 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 4 67 theme lines 816:820 arg1 glycosylation 702:714 the membrane glycosylation 689:714 the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers 689:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 2 68 theme cell 374:377 arg1 signaling 379:387 cell signaling 374:387 cell signaling 374:387 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 1 69 theme women 225:229 arg1 worldwide 231:239 women worldwide 225:239 women worldwide bearing the highest mortality rate among all gynecological cancers 225:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 7 70 gly sialylated 1424:1433 arg1 N-glycans 1435:1443 α 2-6 sialylated N-glycans 1418:1443 α 2-6 sialylated N-glycans 1418:1443 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 71 theme ovarian 1304:1310 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 71 theme ovarian 1304:1310 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 3 72 theme expression 519:528 arg1 regulation 500:509 aberrant regulation 491:509 aberrant regulation of gene expression 491:528 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 11 73 with association 2126:2136 arg1 programming 2154:2164 epigenetic programming 2143:2164 epigenetic programming of their associated synthetic enzymes in ovarian cancer 2143:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 10 74 theme DNA 1905:1907 arg1 hypomethylation 1909:1923 DNA hypomethylation 1905:1923 DNA hypomethylation 1905:1923 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 1 75 theme highest 253:259 arg1 rate 271:274 the highest mortality rate 249:274 the highest mortality rate among all gynecological cancers 249:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 7 76 theme type 1477:1480 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 76 theme type 1477:1480 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 11 77 theme targets 2294:2300 arg1 development 2256:2266 the development 2252:2266 the development of novel anti-glycan drug targets and clinical diagnostic tools 2252:2330 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 4 78 dep lines 816:820 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 78 dep lines 816:820 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 78 dep lines 816:820 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 78 dep lines 816:820 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 78 dep lines 816:820 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 7 79 theme N'-diacetyl-lactosamine 1452:1474 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 79 theme N'-diacetyl-lactosamine 1452:1474 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 9 80 theme potential 1681:1689 arg1 influence 1702:1710 the potential epigenetic influence 1677:1710 the potential epigenetic influence on MGAT3 expression 1677:1730 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 11 81 theme anti-glycan 2277:2287 arg1 targets 2294:2300 novel anti-glycan drug targets 2271:2300 novel anti-glycan drug targets 2271:2300 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 11 82 from enzymes 2196:2202 arg1 cancer 2215:2220 ovarian cancer 2207:2220 ovarian cancer 2207:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 2 83 theme membrane 314:321 arg1 glycans 323:329 Cell membrane glycans 309:329 Cell membrane glycans 309:329 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 11 84 theme epigenetic 2143:2152 arg1 programming 2154:2164 epigenetic programming 2143:2164 epigenetic programming of their associated synthetic enzymes in ovarian cancer 2143:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 7 85 theme N-glycans 1435:1443 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 85 theme N-glycans 1435:1443 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 11 86 theme tools 2326:2330 arg1 development 2256:2266 the development 2252:2266 the development of novel anti-glycan drug targets and clinical diagnostic tools 2252:2330 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 5 87 from chromatography 1015:1028 arg1 carbon 1052:1057 porous graphitized carbon 1033:1057 porous graphitized carbon 1033:1057 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 87 from chromatography 1015:1028 arg1 spectrometry 1105:1116 negative-ion electrospray ionization mass spectrometry 1063:1116 negative-ion electrospray ionization mass spectrometry (ESI-MS) 1063:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 87 from chromatography 1015:1028 arg1 ESI-MS 1119:1124 ESI-MS 1119:1124 ESI-MS 1119:1124 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 9 88 theme MGAT3 1715:1719 arg1 expression 1721:1730 MGAT3 expression 1715:1730 MGAT3 expression 1715:1730 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 11 89 theme clinical 2306:2313 arg1 tools 2326:2330 clinical diagnostic tools 2306:2330 clinical diagnostic tools 2306:2330 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 2 90 theme various 339:345 arg1 signaling 379:387 cell signaling 374:387 cell signaling 374:387 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 2 90 theme various 339:345 arg1 processes 356:364 various cellular processes 339:364 various cellular processes such as cell signaling 339:387 Cell membrane glycans mediate various cellular processes such as cell signaling and become altered during carcinogenesis. 24855066 0 91 theme methylation 135:145 arg1 status 147:152 DNA methylation status 131:152 DNA methylation status 131:152 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 5 92 theme negative-ion 1063:1074 arg1 ESI-MS 1119:1124 ESI-MS 1119:1124 ESI-MS 1119:1124 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 92 theme negative-ion 1063:1074 arg1 spectrometry 1105:1116 negative-ion electrospray ionization mass spectrometry 1063:1116 negative-ion electrospray ionization mass spectrometry (ESI-MS) 1063:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 4 93 theme epithelial 892:901 arg1 cancers 911:917 epithelial ovarian cancers 892:917 epithelial ovarian cancers 892:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 10 94 theme type 1984:1987 arg1 N-glycans 1989:1997 the unique "bisecting GlcNAc" type N-glycans 1954:1997 the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1954:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 5 95 theme ionization 1089:1098 arg1 ESI-MS 1119:1124 ESI-MS 1119:1124 ESI-MS 1119:1124 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 5 95 theme ionization 1089:1098 arg1 spectrometry 1105:1116 negative-ion electrospray ionization mass spectrometry 1063:1116 negative-ion electrospray ionization mass spectrometry (ESI-MS) 1063:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 10 96 from synthesis 1941:1949 arg1 proteins 2015:2022 the membrane proteins 2002:2022 the membrane proteins of ovarian cancer cells 2002:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 11 97 theme substructures 2094:2106 arg1 observation 2061:2071 the observation 2057:2071 the observation of specific N-glycan substructures 2057:2106 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 7 98 theme increased 1398:1406 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 98 theme increased 1398:1406 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 4 99 theme ovarian 733:739 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 99 theme ovarian 733:739 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 99 theme ovarian 733:739 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 99 theme ovarian 733:739 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 99 theme ovarian 733:739 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 99 theme ovarian 733:739 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 10 100 theme GlcNAc 1976:1981 arg1 N-glycans 1989:1997 the unique "bisecting GlcNAc" type N-glycans 1954:1997 the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1954:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 5 101 theme porous 1033:1038 arg1 carbon 1052:1057 porous graphitized carbon 1033:1057 porous graphitized carbon 1033:1057 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 3 102 theme disease 644:650 arg1 progression 624:634 progression 624:634 progression 624:634 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 3 102 theme disease 644:650 arg1 development 608:618 development 608:618 development 608:618 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 11 103 theme specific 2076:2083 arg1 substructures 2094:2106 specific N-glycan substructures 2076:2106 specific N-glycan substructures 2076:2106 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 6 104 theme fragmentation 1211:1223 arg1 patterns 1225:1232 tandem MS fragmentation patterns 1201:1232 tandem MS fragmentation patterns 1201:1232 Glycan structures were characterized based on their molecular masses and tandem MS fragmentation patterns. 24855066 4 105 theme epithelial 749:758 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 105 theme epithelial 749:758 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 105 theme epithelial 749:758 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 105 theme epithelial 749:758 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 105 theme epithelial 749:758 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 105 theme epithelial 749:758 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 10 106 theme membrane 2006:2013 arg1 proteins 2015:2022 the membrane proteins 2002:2022 the membrane proteins of ovarian cancer cells 2002:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 0 107 theme cell 73:76 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 7 108 theme " 1380:1380 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 108 theme " 1380:1380 arg1 N-glycans 1482:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 N'-diacetyl-lactosamine" type N-glycans 1452:1490 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 108 theme " 1380:1380 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 108 theme " 1380:1380 arg1 levels 1408:1413 increased levels 1398:1413 increased levels of α 2-6 sialylated N-glycans and "N 1398:1450 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 4 109 theme membrane 693:700 arg1 glycosylation 702:714 the membrane glycosylation 689:714 the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers 689:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 11 110 theme novel 2271:2275 arg1 targets 2294:2300 novel anti-glycan drug targets 2271:2300 novel anti-glycan drug targets 2271:2300 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 11 111 from programming 2154:2164 arg1 cancer 2215:2220 ovarian cancer 2207:2220 ovarian cancer 2207:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 1 112 theme Epithelial 155:164 arg1 cancer 174:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer 155:179 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 1 112 theme Epithelial 155:164 arg1 cause 206:210 the fifth most common cause 184:210 the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers 184:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 0 113 from glycosylation 9:21 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 10 114 theme cancer 2035:2040 arg1 cells 2042:2046 ovarian cancer cells 2027:2046 ovarian cancer cells 2027:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 9 115 theme methylation 1785:1795 arg1 inhibitor 1797:1805 a DNA methylation inhibitor 1779:1805 a DNA methylation inhibitor 1779:1805 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 9 115 theme methylation 1785:1795 arg1 5-azacytidine 1764:1776 5-azacytidine 1764:1776 5-azacytidine 1764:1776 We further evaluated the potential epigenetic influence on MGAT3 expression by treating the cell lines with 5-azacytidine, a DNA methylation inhibitor. 24855066 8 116 theme transcript 1548:1557 arg1 levels 1559:1564 gene transcript levels 1543:1564 gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR 1543:1653 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 0 117 theme gene 111:114 arg1 expression 116:125 gene expression 111:125 gene expression 111:125 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 5 118 theme membrane 949:956 arg1 glycoproteins 958:970 membrane glycoproteins 949:970 membrane glycoproteins 949:970 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 3 119 theme glycosylation 451:463 arg1 changes 465:471 glycosylation changes 451:471 glycosylation changes 451:471 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 10 120 from N-glycans 1989:1997 arg1 proteins 2015:2022 the membrane proteins 2002:2022 the membrane proteins of ovarian cancer cells 2002:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 11 121 theme enzymes 2196:2202 arg1 programming 2154:2164 epigenetic programming 2143:2164 epigenetic programming of their associated synthetic enzymes in ovarian cancer 2143:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 8 122 theme enzymes 1573:1579 arg1 levels 1559:1564 gene transcript levels 1543:1564 gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR 1543:1653 These N-glycan changes were verified by examining gene transcript levels of the enzymes specific for their synthesis (MGAT3, ST6GAL1, and B4GALNT3) using qRT-PCR. 24855066 4 123 theme HOSE 761:764 arg1 6.3 766:768 HOSE 6.3 761:768 HOSE 6.3 761:768 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 0 124 theme membrane 26:33 arg1 proteins 35:42 membrane proteins 26:42 membrane proteins 26:42 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 11 125 theme associated 2175:2184 arg1 enzymes 2196:2202 their associated synthetic enzymes 2169:2202 their associated synthetic enzymes in ovarian cancer 2169:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 1 126 theme gynecological 286:298 arg1 cancers 300:306 all gynecological cancers 282:306 all gynecological cancers 282:306 Epithelial ovarian cancer is the fifth most common cause of cancer in women worldwide bearing the highest mortality rate among all gynecological cancers. 24855066 10 127 from proteins 2015:2022 arg1 synthesis 1941:1949 the synthesis 1937:1949 the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells 1937:2046 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 7 128 theme cancer 1312:1317 arg1 proteins 1328:1335 the ovarian cancer membrane proteins 1300:1335 the ovarian cancer membrane proteins 1300:1335 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 7 128 theme cancer 1312:1317 arg1 N-glycans 1387:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 the "bisecting N-acetyl-glucosamine" type N-glycans 1345:1395 We identified characteristic glycan features that were unique to the ovarian cancer membrane proteins, namely the "bisecting N-acetyl-glucosamine" type N-glycans, increased levels of α 2-6 sialylated N-glycans and "N,N'-diacetyl-lactosamine" type N-glycans. 24855066 10 129 theme MGAT3 1853:1857 arg1 expression 1859:1868 MGAT3 expression 1853:1868 MGAT3 expression 1853:1868 For the first time, we provide evidence that MGAT3 expression may be epigenetically regulated by DNA hypomethylation, leading to the synthesis of the unique "bisecting GlcNAc" type N-glycans on the membrane proteins of ovarian cancer cells. 24855066 0 130 theme ovarian 58:64 arg1 lines 78:82 epithelial ovarian cancer cell lines 47:82 epithelial ovarian cancer cell lines 47:82 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 5 131 theme nano-liquid 1003:1013 arg1 chromatography 1015:1028 nano-liquid chromatography 1003:1028 nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS) 1003:1125 N-glycans were released from membrane glycoproteins by PNGase F and analyzed using nano-liquid chromatography on porous graphitized carbon and negative-ion electrospray ionization mass spectrometry (ESI-MS). 24855066 11 132 theme ovarian 2207:2213 arg1 cancer 2215:2220 ovarian cancer 2207:2220 ovarian cancer 2207:2220 Linking the observation of specific N-glycan substructures and their complex association with epigenetic programming of their associated synthetic enzymes in ovarian cancer could potentially be used for the development of novel anti-glycan drug targets and clinical diagnostic tools. 24855066 3 133 theme gene 514:517 arg1 expression 519:528 gene expression 514:528 gene expression 514:528 The extent to which glycosylation changes are influenced by aberrant regulation of gene expression is nearly unknown for ovarian cancer and remains crucial in understanding the development and progression of this disease. 24855066 4 134 theme ovarian 796:802 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 134 theme ovarian 796:802 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 134 theme ovarian 796:802 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 134 theme ovarian 796:802 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 134 theme ovarian 796:802 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 134 theme ovarian 796:802 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 0 135 dep glycosylation 9:21 arg1 reflect 103:109 reflect 103:109 reflect gene expression and DNA methylation status 103:152 Specific glycosylation of membrane proteins in epithelial ovarian cancer cell lines: glycan structures reflect gene expression and DNA methylation status. 24855066 4 136 theme cell 811:814 arg1 IGROV1 831:836 IGROV1 831:836 IGROV1 831:836 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 136 theme cell 811:814 arg1 lines 816:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines 719:820 non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3) 719:857 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 136 theme cell 811:814 arg1 A2780 839:843 A2780 839:843 A2780 839:843 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 136 theme cell 811:814 arg1 SKOV 823:826 SKOV 823:826 SKOV 823:826 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 136 theme cell 811:814 arg1 OVCAR 850:854 OVCAR 850:854 OVCAR 850:854 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 24855066 4 136 theme cell 811:814 arg1 histotype 876:884 the most common histotype 860:884 the most common histotype among epithelial ovarian cancers 860:917 To address this effect, we analyzed the membrane glycosylation of non-cancerous ovarian surface epithelial (HOSE 6.3 and HOSE 17.1) and serous ovarian cancer cell lines (SKOV 3, IGROV1, A2780, and OVCAR 3), the most common histotype among epithelial ovarian cancers. 27873218 3 0 theme Bikunin 593:599 arg1 proteoglycan 601:612 the Bikunin proteoglycan 589:612 the Bikunin proteoglycan of human urine samples 589:635 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 1 1 theme site-specific 307:319 arg1 analysis 321:328 site-specific analysis 307:328 site-specific analysis of protein N- and O-glycosylations 307:363 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 3 2 theme urine 623:627 arg1 samples 629:635 human urine samples 617:635 human urine samples 617:635 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 6 3 theme structure 1290:1298 arg1 GalNAc 1261:1266 the GalNAc 1257:1266 the GalNAc of the hexasaccharide structure 1257:1298 Sulfate groups were pinpointed to both Gal residues and to the GalNAc of the hexasaccharide structure. 27873218 9 4 theme variants 1626:1633 arg1 beneficial 1645:1654 beneficial 1645:1654 beneficial 1645:1654 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 9 4 theme variants 1626:1633 arg1 realization 1594:1604 The realization 1590:1604 The realization of these structural variants 1590:1633 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 3 5 theme ΔHexAGalNAcGlcAGalGalXyl-O-Ser 642:671 arg1 structure 688:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 7 6 theme substituents 1472:1483 arg1 analysis 1452:1459 complementary structural analysis 1427:1459 complementary structural analysis of sulfate substituents of CS-glycopeptides 1427:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 10 7 gly glycopeptides 1987:1999 arg2 glycopeptides 1987:1999 sulfated glycopeptides 1978:1999 sulfated glycopeptides 1978:1999 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 10 8 theme sulfated 1978:1985 arg1 glycopeptides 1987:1999 sulfated glycopeptides 1978:1999 sulfated glycopeptides 1978:1999 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 1 9 theme protease 264:271 arg1 digests 273:279 protease digests 264:279 protease digests of glycoproteins 264:296 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 4 10 theme sulfate 819:825 arg1 residues 838:845 sulfate attachment residues 819:845 sulfate attachment residues 819:845 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 5 11 theme modifications 1183:1195 arg1 assignment 1161:1170 the assignment 1157:1170 the assignment of sulfate modifications 1157:1195 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 5 12 theme Na+ 1053:1055 arg1 ions 1057:1060 Na+ ions 1053:1060 Na+ ions 1053:1060 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 3 13 theme 0-3 732:734 arg1 sulfate 736:742 0-3 sulfate 732:742 0-3 sulfate 732:742 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 5 14 theme sulfate 1014:1020 arg1 instability 1022:1032 the well-recognized sulfate instability 994:1032 the well-recognized sulfate instability 994:1032 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 2 15 gly glycopeptides 487:499 arg2 CS-glycopeptides 502:517 CS-glycopeptides 502:517 CS-glycopeptides 502:517 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 2 15 gly glycopeptides 487:499 arg2 glycopeptides 487:499 chondroitin sulfate-containing glycopeptides 456:499 chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS 456:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 0 16 theme Ion-Pairing 97:107 arg1 LC-MS/MS 127:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 4 17 theme protonated 852:861 arg1 precursors 863:872 protonated precursors 852:872 protonated precursors 852:872 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 0 18 theme Positive 113:120 arg1 LC-MS/MS 127:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 9 19 theme linkage 1715:1721 arg1 region 1723:1728 the CS linkage region 1708:1728 the CS linkage region 1708:1728 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 1 20 theme chromatography-tandem 161:181 arg1 spectrometry 188:199 liquid chromatography-tandem mass spectrometry 154:199 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 154:210 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 20 theme chromatography-tandem 161:181 arg1 LC-MS/MS 202:209 LC-MS/MS 202:209 LC-MS/MS 202:209 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 21 dep Purification 137:148 arg1 characterization 212:227 characterization 212:227 characterization 212:227 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 7 22 theme CS-glycopeptides 1488:1503 arg1 substituents 1472:1483 sulfate substituents 1464:1483 sulfate substituents of CS-glycopeptides 1464:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 2 23 theme positive 526:533 arg1 LC-MS/MS 540:547 positive mode LC-MS/MS 526:547 positive mode LC-MS/MS 526:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 8 24 theme considerable 1529:1540 arg1 heterogeneity 1542:1554 a considerable heterogeneity 1527:1554 a considerable heterogeneity of the bikunin CS linkage region 1527:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 1 25 theme protein 333:339 arg1 N- 341:342 protein N- 333:342 protein N- 333:342 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 6 26 theme Sulfate 1198:1204 arg1 groups 1206:1211 Sulfate groups 1198:1211 Sulfate groups 1198:1211 Sulfate groups were pinpointed to both Gal residues and to the GalNAc of the hexasaccharide structure. 27873218 3 27 contain had 638:640 arg2 structure 688:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 3 27 contain had 638:640 arg1 CS-glycopeptides 554:569 The CS-glycopeptides 550:569 The CS-glycopeptides 550:569 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 4 28 theme high-energy 930:940 arg1 HCD 974:976 HCD 974:976 HCD 974:976 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 4 28 theme high-energy 930:940 arg1 dissociation 960:971 high-energy collision induced dissociation 930:971 high-energy collision induced dissociation (HCD) 930:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 0 29 theme Glycan 20:25 arg1 Structures 27:36 Glycan Structures 20:36 Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS 20:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 5 30 theme sodiated 1070:1077 arg1 precursors 1079:1088 sodiated precursors 1070:1088 sodiated precursors 1070:1088 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 1 31 theme mass 183:186 arg1 spectrometry 188:199 liquid chromatography-tandem mass spectrometry 154:199 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 154:210 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 31 theme mass 183:186 arg1 LC-MS/MS 202:209 LC-MS/MS 202:209 LC-MS/MS 202:209 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 10 32 theme combined 1830:1837 arg1 use 1839:1841 the combined use 1826:1841 the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis 1826:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 8 33 theme bikunin 1563:1569 arg1 region 1582:1587 the bikunin CS linkage region 1559:1587 the bikunin CS linkage region 1559:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 2 34 theme sulfate-containing 468:485 arg1 CS-glycopeptides 502:517 CS-glycopeptides 502:517 CS-glycopeptides 502:517 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 2 34 theme sulfate-containing 468:485 arg1 glycopeptides 487:499 chondroitin sulfate-containing glycopeptides 456:499 chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS 456:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 9 35 theme CS 1766:1767 arg1 biosynthesis 1750:1761 the biosynthesis 1746:1761 the biosynthesis of CS 1746:1767 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 9 36 from beneficial 1645:1654 arg1 studies 1659:1665 studies 1659:1665 studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins 1659:1817 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 7 37 theme structural 1441:1450 arg1 analysis 1452:1459 complementary structural analysis 1427:1459 complementary structural analysis of sulfate substituents of CS-glycopeptides 1427:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 3 38 theme phosphate 752:760 arg1 groups 762:767 0-3 sulfate and 0-1 phosphate groups 732:767 0-3 sulfate and 0-1 phosphate groups 732:767 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 8 39 theme linkage 1574:1580 arg1 region 1582:1587 the bikunin CS linkage region 1559:1587 the bikunin CS linkage region 1559:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 10 40 theme precursors 1870:1879 arg1 use 1839:1841 the combined use 1826:1841 the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis 1826:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 0 41 theme Sulfate-Glycopeptides 53:73 arg1 Structures 27:36 Glycan Structures 20:36 Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS 20:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 8 42 gly heterogeneity 1542:1554 arg1 region 1582:1587 the bikunin CS linkage region 1559:1587 the bikunin CS linkage region 1559:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 9 43 theme potential 1773:1781 arg1 interactions 1783:1794 potential interactions 1773:1794 potential interactions to CS binding proteins 1773:1817 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 10 44 theme fragmentation 1903:1915 arg1 analysis 1917:1924 positive mode HCD fragmentation analysis 1885:1924 positive mode HCD fragmentation analysis 1885:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 4 45 theme groups 917:922 arg1 fragmentation 892:904 extensive fragmentation 882:904 extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD) 882:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 2 46 theme chondroitinase 419:432 arg1 ABC 434:436 chondroitinase ABC 419:436 chondroitinase ABC 419:436 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 1 47 theme glycoproteins 284:296 arg1 digests 273:279 protease digests 264:279 protease digests of glycoproteins 264:296 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 9 48 theme binding 1802:1808 arg1 proteins 1810:1817 CS binding proteins 1799:1817 CS binding proteins 1799:1817 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 6 49 theme Gal 1237:1239 arg1 residues 1241:1248 both Gal residues 1232:1248 both Gal residues 1232:1248 Sulfate groups were pinpointed to both Gal residues and to the GalNAc of the hexasaccharide structure. 27873218 10 50 theme positive 1885:1892 arg1 analysis 1917:1924 positive mode HCD fragmentation analysis 1885:1924 positive mode HCD fragmentation analysis 1885:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 7 51 from prominent 1378:1386 arg1 spectra 1403:1409 the HCD-MS2 spectra 1391:1409 the HCD-MS2 spectra 1391:1409 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 1 52 gly glycopeptides 232:244 arg2 glycopeptides 232:244 glycopeptides 232:244 glycopeptides 232:244 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 4 53 theme induced 952:958 arg1 HCD 974:976 HCD 974:976 HCD 974:976 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 4 53 theme induced 952:958 arg1 dissociation 960:971 high-energy collision induced dissociation 930:971 high-energy collision induced dissociation (HCD) 930:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 3 54 theme samples 629:635 arg1 proteoglycan 601:612 the Bikunin proteoglycan 589:612 the Bikunin proteoglycan of human urine samples 589:635 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 10 55 theme additional 1956:1965 arg1 classes 1967:1973 additional classes 1956:1973 additional classes of sulfated glycopeptides 1956:1999 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 3 56 theme human 617:621 arg1 samples 629:635 human urine samples 617:635 human urine samples 617:635 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 7 57 from spectra 1403:1409 arg1 prominent 1378:1386 prominent 1378:1386 prominent 1378:1386 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 7 57 from spectra 1403:1409 arg1 intensities 1305:1315 The intensities 1301:1315 The intensities of protonated and sodiated saccharide oxonium ions 1301:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 7 58 theme ions 1363:1366 arg1 prominent 1378:1386 prominent 1378:1386 prominent 1378:1386 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 7 58 theme ions 1363:1366 arg1 intensities 1305:1315 The intensities 1301:1315 The intensities of protonated and sodiated saccharide oxonium ions 1301:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 3 59 theme hexasaccharide 673:686 arg1 structure 688:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure 642:696 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 9 60 theme structural 1615:1624 arg1 variants 1626:1633 these structural variants 1609:1633 these structural variants 1609:1633 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 9 61 from studies 1659:1665 arg1 beneficial 1645:1654 beneficial 1645:1654 beneficial 1645:1654 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 9 61 from studies 1659:1665 arg1 realization 1594:1604 The realization 1590:1604 The realization of these structural variants 1590:1633 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 6 62 theme hexasaccharide 1275:1288 arg1 structure 1290:1298 the hexasaccharide structure 1271:1298 the hexasaccharide structure 1271:1298 Sulfate groups were pinpointed to both Gal residues and to the GalNAc of the hexasaccharide structure. 27873218 4 63 theme collision 942:950 arg1 HCD 974:976 HCD 974:976 HCD 974:976 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 4 63 theme collision 942:950 arg1 dissociation 960:971 high-energy collision induced dissociation 930:971 high-energy collision induced dissociation (HCD) 930:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 10 64 theme glycopeptides 1987:1999 arg1 classes 1967:1973 additional classes 1956:1973 additional classes of sulfated glycopeptides 1956:1999 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 1 65 theme glycopeptides 232:244 arg1 spectrometry 188:199 liquid chromatography-tandem mass spectrometry 154:199 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 154:210 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 65 theme glycopeptides 232:244 arg1 Purification 137:148 Purification 137:148 Purification 137:148 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 65 theme glycopeptides 232:244 arg1 LC-MS/MS 202:209 LC-MS/MS 202:209 LC-MS/MS 202:209 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 7 66 theme sodiated 1335:1342 arg1 ions 1363:1366 protonated and sodiated saccharide oxonium ions 1320:1366 protonated and sodiated saccharide oxonium ions 1320:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 11 67 theme Graphical 2002:2010 arg1 Abstract 2012:2019 Graphical Abstract 2002:2019 Graphical Abstract 2002:2019 Graphical Abstract ᅟ. 27873218 5 68 theme sulfate 1175:1181 arg1 modifications 1183:1195 sulfate modifications 1175:1195 sulfate modifications 1175:1195 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 3 69 theme sulfate 736:742 arg1 groups 762:767 0-3 sulfate and 0-1 phosphate groups 732:767 0-3 sulfate and 0-1 phosphate groups 732:767 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 1 70 theme liquid 154:159 arg1 spectrometry 188:199 liquid chromatography-tandem mass spectrometry 154:199 liquid chromatography-tandem mass spectrometry (LC-MS/MS) 154:210 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 1 70 theme liquid 154:159 arg1 LC-MS/MS 202:209 LC-MS/MS 202:209 LC-MS/MS 202:209 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 4 71 theme attachment 827:836 arg1 residues 838:845 sulfate attachment residues 819:845 sulfate attachment residues 819:845 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 5 72 theme well-recognized 998:1012 arg1 instability 1022:1032 the well-recognized sulfate instability 994:1032 the well-recognized sulfate instability 994:1032 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 2 73 gly CS-glycopeptides 502:517 arg2 CS-glycopeptides 502:517 CS-glycopeptides 502:517 CS-glycopeptides 502:517 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 2 73 gly CS-glycopeptides 502:517 arg2 glycopeptides 487:499 chondroitin sulfate-containing glycopeptides 456:499 chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS 456:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 7 74 theme saccharide 1344:1353 arg1 ions 1363:1366 protonated and sodiated saccharide oxonium ions 1320:1366 protonated and sodiated saccharide oxonium ions 1320:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 7 75 theme oxonium 1355:1361 arg1 ions 1363:1366 protonated and sodiated saccharide oxonium ions 1320:1366 protonated and sodiated saccharide oxonium ions 1320:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 0 76 theme Mode 122:125 arg1 LC-MS/MS 127:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Sodium Ion-Pairing and Positive Mode LC-MS/MS 90:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 9 77 theme CS 1712:1713 arg1 region 1723:1728 the CS linkage region 1708:1728 the CS linkage region 1708:1728 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 10 78 theme protonated 1846:1855 arg1 precursors 1870:1879 protonated and sodiated precursors 1846:1879 protonated and sodiated precursors 1846:1879 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 7 79 theme protonated 1320:1329 arg1 ions 1363:1366 protonated and sodiated saccharide oxonium ions 1320:1366 protonated and sodiated saccharide oxonium ions 1320:1366 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 7 80 gly CS-glycopeptides 1488:1503 arg2 CS-glycopeptides 1488:1503 CS-glycopeptides 1488:1503 CS-glycopeptides 1488:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 1 81 gly glycoproteins 284:296 arg1 glycoproteins 284:296 glycoproteins 284:296 glycoproteins 284:296 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 9 82 theme region 1723:1728 arg1 importance 1694:1703 the importance 1690:1703 the importance of the CS linkage region 1690:1728 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 7 83 theme sulfate 1464:1470 arg1 substituents 1472:1483 sulfate substituents 1464:1483 sulfate substituents of CS-glycopeptides 1464:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 0 84 theme Structures 27:36 arg1 Characterization 0:15 Characterization 0:15 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 0:135 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 1 85 theme N- 341:342 arg1 analysis 321:328 site-specific analysis 307:328 site-specific analysis of protein N- and O-glycosylations 307:363 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 4 86 theme extensive 882:890 arg1 fragmentation 892:904 extensive fragmentation 882:904 extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD) 882:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 3 87 gly CS-glycopeptides 554:569 arg2 CS-glycopeptides 554:569 The CS-glycopeptides 550:569 The CS-glycopeptides 550:569 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 10 88 theme HCD 1899:1901 arg1 analysis 1917:1924 positive mode HCD fragmentation analysis 1885:1924 positive mode HCD fragmentation analysis 1885:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 0 89 theme Chondroitin 41:51 arg1 Sulfate-Glycopeptides 53:73 Chondroitin Sulfate-Glycopeptides 41:73 Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS 41:134 Characterization of Glycan Structures of Chondroitin Sulfate-Glycopeptides Facilitated by Sodium Ion-Pairing and Positive Mode LC-MS/MS. 27873218 1 90 theme O-glycosylations 348:363 arg1 analysis 321:328 site-specific analysis 307:328 site-specific analysis of protein N- and O-glycosylations 307:363 Purification and liquid chromatography-tandem mass spectrometry (LC-MS/MS) characterization of glycopeptides, originating from protease digests of glycoproteins, enables site-specific analysis of protein N- and O-glycosylations. 27873218 7 91 theme complementary 1427:1439 arg1 analysis 1452:1459 complementary structural analysis 1427:1459 complementary structural analysis of sulfate substituents of CS-glycopeptides 1427:1503 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 3 92 theme 0-1 748:750 arg1 phosphate 752:760 0-1 phosphate 748:760 0-1 phosphate 748:760 The CS-glycopeptides, originating from the Bikunin proteoglycan of human urine samples, had ΔHexAGalNAcGlcAGalGalXyl-O-Ser hexasaccharide structure and were further substituted with 0-3 sulfate and 0-1 phosphate groups. 27873218 2 93 theme chondroitin 456:466 arg1 CS-glycopeptides 502:517 CS-glycopeptides 502:517 CS-glycopeptides 502:517 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 2 93 theme chondroitin 456:466 arg1 glycopeptides 487:499 chondroitin sulfate-containing glycopeptides 456:499 chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS 456:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 2 94 theme mode 535:538 arg1 LC-MS/MS 540:547 positive mode LC-MS/MS 526:547 positive mode LC-MS/MS 526:547 We have described a protocol to enrich, hydrolyze by chondroitinase ABC, and characterize chondroitin sulfate-containing glycopeptides (CS-glycopeptides) using positive mode LC-MS/MS. 27873218 8 95 theme CS 1571:1572 arg1 region 1582:1587 the bikunin CS linkage region 1559:1587 the bikunin CS linkage region 1559:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 10 96 theme sodiated 1861:1868 arg1 precursors 1870:1879 protonated and sodiated precursors 1846:1879 protonated and sodiated precursors 1846:1879 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 27873218 5 97 theme sulfate 1107:1113 arg1 groups 1115:1120 sulfate groups 1107:1120 sulfate groups 1107:1120 To circumvent the well-recognized sulfate instability, we now introduced Na+ ions to form sodiated precursors, which protected sulfate groups from decomposition and facilitated the assignment of sulfate modifications. 27873218 8 98 theme region 1582:1587 arg1 heterogeneity 1542:1554 a considerable heterogeneity 1527:1554 a considerable heterogeneity of the bikunin CS linkage region 1527:1587 We have demonstrated a considerable heterogeneity of the bikunin CS linkage region. 27873218 7 99 theme HCD-MS2 1395:1401 arg1 spectra 1403:1409 the HCD-MS2 spectra 1391:1409 the HCD-MS2 spectra 1391:1409 The intensities of protonated and sodiated saccharide oxonium ions were very prominent in the HCD-MS2 spectra, which provided complementary structural analysis of sulfate substituents of CS-glycopeptides. 27873218 9 100 theme CS 1799:1800 arg1 proteins 1810:1817 CS binding proteins 1799:1817 CS binding proteins 1799:1817 The realization of these structural variants should be beneficial in studies aimed at investigating the importance of the CS linkage region with regards to the biosynthesis of CS and potential interactions to CS binding proteins. 27873218 4 101 theme sulfate 909:915 arg1 groups 917:922 sulfate groups 909:922 sulfate groups using high-energy collision induced dissociation (HCD) 909:977 However, it was not possible to exactly pinpoint sulfate attachment residues, for protonated precursors, due to extensive fragmentation of sulfate groups using high-energy collision induced dissociation (HCD). 27873218 10 102 theme mode 1894:1897 arg1 analysis 1917:1924 positive mode HCD fragmentation analysis 1885:1924 positive mode HCD fragmentation analysis 1885:1924 Also, the combined use of protonated and sodiated precursors for positive mode HCD fragmentation analysis will likely become useful for additional classes of sulfated glycopeptides. 28669180 6 0 from process 1023:1029 arg1 present 995:1001 present 995:1001 present 995:1001 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 1 theme fermentation 1010:1021 arg1 process 1023:1029 the fermentation process 1006:1029 the fermentation process 1006:1029 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 9 2 attach released 1586:1593 arg1 walls 1609:1613 the cell walls 1600:1613 the cell walls of Malvar grapes and the carbohydrates released from each yeast species 1600:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 9 2 attach released 1586:1593 arg2 polysaccharides 1549:1563 polysaccharides 1549:1563 polysaccharides 1549:1563 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 8 3 theme delbrueckii 1421:1431 arg1 CLI 1433:1435 T. delbrueckii CLI 918 1418:1439 T. delbrueckii CLI 918 1418:1439 Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. 28669180 6 4 from comparison 1034:1043 arg1 present 995:1001 present 995:1001 present 995:1001 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 7 5 from rich 1167:1170 arg1 galactose 1189:1197 galactose 1189:1197 galactose 1189:1197 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 7 5 from rich 1167:1170 arg1 arabinose 1175:1183 arabinose 1175:1183 arabinose 1175:1183 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 0 6 theme delbrueckii 90:100 arg1 Yeasts 154:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 7 7 theme pure 1227:1230 arg1 cultures 1232:1239 pure cultures 1227:1239 pure cultures of S. cerevisiae 1227:1256 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 9 8 theme grapes 1625:1630 arg1 walls 1609:1613 the cell walls 1600:1613 the cell walls of Malvar grapes and the carbohydrates released from each yeast species 1600:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 7 9 from arabinose 1175:1183 arg1 rich 1167:1170 rich 1167:1170 rich 1167:1170 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 8 10 theme cerevisiae 1466:1475 arg1 CLI 1477:1479 S. cerevisiae CLI 889 1463:1483 S. cerevisiae CLI 889 1463:1483 Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. 28669180 6 11 attach present 995:1001 arg2 T. 976:977 T. 976:977 T. 976:977 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 11 attach present 995:1001 arg1 comparison 1034:1043 comparison 1034:1043 comparison with 1034:1048 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 11 attach present 995:1001 arg1 process 1023:1029 the fermentation process 1006:1029 the fermentation process 1006:1029 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 12 attach present 1091:1097 arg2 cultures 1060:1067 pure cultures 1055:1067 pure cultures of S. cerevisiae 1055:1084 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 12 attach present 1091:1097 arg1 process 1118:1124 the fermenation process 1102:1124 the fermenation process 1102:1124 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 1 13 theme pure 279:282 arg1 cultures 307:314 pure, mixed, and sequential cultures 279:314 pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 279:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 5 14 theme inoculation 799:809 arg1 species 779:785 Yeast species 773:785 Yeast species 773:785 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 5 14 theme inoculation 799:809 arg1 type 791:794 type 791:794 type of inoculation 791:809 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 1 15 theme cerevisiae 371:380 arg1 yeasts 397:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 6 16 from process 1118:1124 arg1 present 1091:1097 present 1091:1097 present 1091:1097 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 1 17 with cultures 307:314 arg1 CLI 345:347 Torulaspora delbrueckii CLI 918 321:351 Torulaspora delbrueckii CLI 918 321:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 3 18 theme wine 578:581 arg1 linkages 598:605 wine polysaccharide linkages 578:605 wine polysaccharide linkages 578:605 Glycosyl composition and wine polysaccharide linkages were determined by GC-EI-MS chromatography. 28669180 6 19 theme cerevisiae 1075:1084 arg1 cultures 1060:1067 pure cultures 1055:1067 pure cultures of S. cerevisiae 1055:1084 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 0 20 theme cerevisiae 128:137 arg1 Yeasts 154:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 1 21 theme CLI 382:384 arg1 yeasts 397:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 6 22 from mannoproteins 907:919 arg1 residues 893:900 Mannose residues 885:900 Mannose residues from mannoproteins 885:919 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 8 23 theme T. 1418:1419 arg1 CLI 1433:1435 T. delbrueckii CLI 918 1418:1439 T. delbrueckii CLI 918 1418:1439 Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. 28669180 3 24 theme Glycosyl 553:560 arg1 composition 562:572 Glycosyl composition 553:572 Glycosyl composition 553:572 Glycosyl composition and wine polysaccharide linkages were determined by GC-EI-MS chromatography. 28669180 7 25 theme greater 1209:1215 arg1 values 1217:1222 greater values 1209:1222 greater values 1209:1222 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 2 26 theme high-resolution 506:520 arg1 chromatography 537:550 high-resolution size-exclusion chromatography 506:550 high-resolution size-exclusion chromatography 506:550 Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. 28669180 5 27 theme carbohydrate 845:856 arg1 composition 858:868 wine carbohydrate composition 840:868 wine carbohydrate composition 840:868 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 4 28 theme Molar-mass 651:660 arg1 distributions 662:674 Molar-mass distributions 651:674 Molar-mass distributions 651:674 Molar-mass distributions were determined by SEC-MALLS, and intrinsic viscosity was determined by differential viscometer. 28669180 2 29 theme size-exclusion 522:535 arg1 chromatography 537:550 high-resolution size-exclusion chromatography 506:550 high-resolution size-exclusion chromatography 506:550 Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. 28669180 9 30 theme each 1668:1671 arg1 species 1679:1685 each yeast species 1668:1685 each yeast species 1668:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 1 31 theme native 390:395 arg1 yeasts 397:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Saccharomyces cerevisiae CLI 889 native yeasts 357:402 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 9 32 theme important 1517:1525 arg1 information 1527:1537 important information 1517:1537 important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species 1517:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 7 33 from galactose 1189:1197 arg1 rich 1167:1170 rich 1167:1170 rich 1167:1170 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 5 34 contain have 811:814 arg1 species 779:785 Yeast species 773:785 Yeast species 773:785 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 5 34 contain have 811:814 arg2 impact 830:835 a significant impact 816:835 a significant impact 816:835 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 5 34 contain have 811:814 arg1 type 791:794 type 791:794 type of inoculation 791:809 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 1 35 theme mixed 285:289 arg1 cultures 307:314 pure, mixed, and sequential cultures 279:314 pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 279:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 6 36 from present 1091:1097 arg1 process 1118:1124 the fermenation process 1102:1124 the fermenation process 1102:1124 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 0 37 dep delbrueckii 90:100 arg1 CLI 102:104 CLI 918 102:108 Torulaspora delbrueckii CLI 918 78:108 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 8 38 theme structural 1361:1370 arg1 differences 1372:1382 structural differences 1361:1382 structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889 1361:1483 Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. 28669180 5 39 theme Yeast 773:777 arg1 species 779:785 Yeast species 773:785 Yeast species 773:785 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 3 40 theme polysaccharide 583:596 arg1 linkages 598:605 wine polysaccharide linkages 578:605 wine polysaccharide linkages 578:605 Glycosyl composition and wine polysaccharide linkages were determined by GC-EI-MS chromatography. 28669180 9 41 theme cell 1604:1607 arg1 walls 1609:1613 the cell walls 1600:1613 the cell walls of Malvar grapes and the carbohydrates released from each yeast species 1600:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 6 42 from cultures 961:968 arg1 predominant 940:950 predominant 940:950 predominant 940:950 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 0 43 theme CLI 139:141 arg1 Yeasts 154:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 9 44 theme yeast 1673:1677 arg1 species 1679:1685 each yeast species 1668:1685 each yeast species 1668:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 6 45 located present 995:1001 arg2 T. 976:977 T. 976:977 T. 976:977 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 45 located present 995:1001 arg1 comparison 1034:1043 comparison 1034:1043 comparison with 1034:1048 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 45 located present 995:1001 arg1 process 1023:1029 the fermentation process 1006:1029 the fermentation process 1006:1029 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 2 46 theme white 476:480 arg1 wines 482:486 different white wines 466:486 different white wines 466:486 Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. 28669180 1 47 theme sequential 296:305 arg1 cultures 307:314 pure, mixed, and sequential cultures 279:314 pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 279:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 7 48 theme rich 1167:1170 arg1 polysaccharides 1151:1165 polysaccharides 1151:1165 polysaccharides rich in arabinose and galactose 1151:1197 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 2 49 from wines 482:486 arg1 fractions 451:459 Both fractions 446:459 Both fractions from different white wines 446:486 Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. 28669180 6 50 theme pure 1055:1058 arg1 cultures 1060:1067 pure cultures 1055:1067 pure cultures of S. cerevisiae 1055:1084 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 51 dep T. 976:977 arg1 delbrueckii 979:989 T. delbrueckii 976:989 T. delbrueckii 976:989 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 2 52 theme different 466:474 arg1 wines 482:486 different white wines 466:486 different white wines 466:486 Both fractions from different white wines were separated by high-resolution size-exclusion chromatography. 28669180 0 53 theme Native 147:152 arg1 Yeasts 154:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts 78:159 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 6 54 theme fermenation 1106:1116 arg1 process 1118:1124 the fermenation process 1102:1124 the fermenation process 1102:1124 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 9 55 theme Malvar 1618:1623 arg1 grapes 1625:1630 Malvar grapes 1618:1630 Malvar grapes 1618:1630 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 6 56 from predominant 940:950 arg1 cultures 961:968 those cultures 955:968 those cultures 955:968 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 57 theme Mannose 885:891 arg1 residues 893:900 Mannose residues 885:900 Mannose residues from mannoproteins 885:919 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 0 58 dep Vinos 172:176 arg1 de 178:179 Vinos de 172:179 D.O. "Vinos de Madrid" 166:187 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 0 58 dep Vinos 172:176 arg1 Madrid 181:186 Madrid 181:186 Madrid 181:186 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 7 59 theme cerevisiae 1247:1256 arg1 cultures 1232:1239 pure cultures 1227:1239 pure cultures of S. cerevisiae 1227:1256 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 1 60 theme Torulaspora 321:331 arg1 CLI 345:347 Torulaspora delbrueckii CLI 918 321:351 Torulaspora delbrueckii CLI 918 321:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 9 61 attach released 1654:1661 arg1 species 1679:1685 each yeast species 1668:1685 each yeast species 1668:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 9 61 attach released 1654:1661 arg2 carbohydrates 1640:1652 the carbohydrates 1636:1652 the carbohydrates released from each yeast species 1636:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 9 62 dep polysaccharides 1549:1563 arg1 the 1545:1547 the 1545:1547 the 1545:1547 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 3 63 theme GC-EI-MS 626:633 arg1 chromatography 635:648 GC-EI-MS chromatography 626:648 GC-EI-MS chromatography 626:648 Glycosyl composition and wine polysaccharide linkages were determined by GC-EI-MS chromatography. 28669180 6 64 attach predominant 940:950 arg2 residues 893:900 Mannose residues 885:900 Mannose residues from mannoproteins 885:919 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 64 attach predominant 940:950 arg1 cultures 961:968 those cultures 955:968 those cultures 955:968 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 1 65 theme delbrueckii 333:343 arg1 CLI 345:347 Torulaspora delbrueckii CLI 918 321:351 Torulaspora delbrueckii CLI 918 321:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 7 66 theme Galactose 1127:1135 arg1 residues 1137:1144 Galactose residues 1127:1144 Galactose residues from polysaccharides rich in arabinose and galactose 1127:1197 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 6 67 from present 995:1001 arg1 comparison 1034:1043 comparison 1034:1043 comparison with 1034:1048 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 67 from present 995:1001 arg1 process 1023:1029 the fermentation process 1006:1029 the fermentation process 1006:1029 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 0 68 dep D.O. 166:169 arg1 Vinos 172:176 Vinos 172:176 Vinos 172:176 Polysaccharides and Oligosaccharides Produced on Malvar Wines Elaborated with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 Native Yeasts from D.O. "Vinos de Madrid". 28669180 1 69 dep D.O. 409:412 arg1 Vinos 415:419 Vinos 415:419 Vinos 415:419 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 1 70 attach released 227:234 arg1 D.O. 409:412 D.O. 409:412 D.O. 409:412 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 1 70 attach released 227:234 arg2 Polysaccharides 190:204 Polysaccharides 190:204 Polysaccharides 190:204 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 6 71 located present 1091:1097 arg2 cultures 1060:1067 pure cultures 1055:1067 pure cultures of S. cerevisiae 1055:1084 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 6 71 located present 1091:1097 arg1 process 1118:1124 the fermenation process 1102:1124 the fermenation process 1102:1124 Mannose residues from mannoproteins were significantly predominant in those cultures where T. delbrueckii was present in the fermentation process in comparison with when pure cultures of S. cerevisiae were present in the fermenation process. 28669180 5 72 theme wine 840:843 arg1 composition 858:868 wine carbohydrate composition 840:868 wine carbohydrate composition 840:868 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 9 73 theme carbohydrates 1640:1652 arg1 walls 1609:1613 the cell walls 1600:1613 the cell walls of Malvar grapes and the carbohydrates released from each yeast species 1600:1685 These findings help to provide important information about the polysaccharides and oligosaccharides released from the cell walls of Malvar grapes and the carbohydrates released from each yeast species. 28669180 7 74 attach presented 1199:1207 arg1 cultures 1232:1239 pure cultures 1227:1239 pure cultures of S. cerevisiae 1227:1256 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 7 74 attach presented 1199:1207 arg2 residues 1137:1144 Galactose residues 1127:1144 Galactose residues from polysaccharides rich in arabinose and galactose 1127:1197 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 1 75 theme Malvar 241:246 arg1 wines 254:258 Malvar white wines 241:258 Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 241:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 5 76 theme significant 818:828 arg1 impact 830:835 a significant impact 816:835 a significant impact 816:835 Yeast species and type of inoculation have a significant impact on wine carbohydrate composition and structure. 28669180 8 77 theme S. 1463:1464 arg1 CLI 1477:1479 S. cerevisiae CLI 889 1463:1483 S. cerevisiae CLI 889 1463:1483 Moreover, we reported structural differences between mannoproteins released by T. delbrueckii CLI 918 and those released by S. cerevisiae CLI 889. 28669180 4 78 theme differential 748:759 arg1 viscometer 761:770 differential viscometer 748:770 differential viscometer 748:770 Molar-mass distributions were determined by SEC-MALLS, and intrinsic viscosity was determined by differential viscometer. 28669180 4 79 theme intrinsic 710:718 arg1 viscosity 720:728 intrinsic viscosity 710:728 intrinsic viscosity 710:728 Molar-mass distributions were determined by SEC-MALLS, and intrinsic viscosity was determined by differential viscometer. 28669180 1 80 theme white 248:252 arg1 wines 254:258 Malvar white wines 241:258 Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 241:351 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 1 81 dep Vinos 415:419 arg1 de 421:422 Vinos de 415:422 D.O. "Vinos de Madrid" 409:430 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 1 81 dep Vinos 415:419 arg1 Madrid 424:429 Madrid 424:429 Madrid 424:429 Polysaccharides and oligosaccharides released into Malvar white wines elaborated through pure, mixed, and sequential cultures with Torulaspora delbrueckii CLI 918 and Saccharomyces cerevisiae CLI 889 native yeasts from D.O. "Vinos de Madrid" were studied. 28669180 7 82 theme fewer 1298:1302 arg1 mannoproteins 1304:1316 fewer mannoproteins 1298:1316 fewer mannoproteins 1298:1316 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 28669180 7 83 from polysaccharides 1151:1165 arg1 residues 1137:1144 Galactose residues 1127:1144 Galactose residues from polysaccharides rich in arabinose and galactose 1127:1197 Galactose residues from polysaccharides rich in arabinose and galactose presented greater values in pure cultures of S. cerevisiae, indicating that S. cerevisiae released fewer mannoproteins than T. delbrueckii. 27748959 6 0 theme α-1,3- 921:926 arg1 MGAT1 984:988 MGAT1 984:988 MGAT1 984:988 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 0 theme α-1,3- 921:926 arg1 beta-1,2-N-acetylglucosaminyltransferase 942:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase 894:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1) 894:989 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 11 1 theme MGAT1 2085:2089 arg1 level 2076:2080 the level 2072:2080 the level of MGAT1 2072:2089 Importantly, the level of MGAT1 was reduced, because likely MGAT1, contrary to Man-I and Man-II, is giantin sensitive. 27748959 6 2 theme mannosyl 911:918 arg1 MGAT1 984:988 MGAT1 984:988 MGAT1 984:988 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 2 theme mannosyl 911:918 arg1 beta-1,2-N-acetylglucosaminyltransferase 942:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase 894:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1) 894:989 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 3 gly -glycoprotein 928:940 arg1 -glycoprotein 928:940 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase 894:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1) 894:989 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 9 4 theme GTPase 1706:1711 arg1 dysfunction 1658:1668 dysfunction 1658:1668 dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase 1658:1711 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 5 dep results 1647:1653 arg1 followed 1713:1720 followed 1713:1720 followed by a deficiency in COPI vesicles at the Golgi 1713:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 10 6 theme Silencing 1769:1777 arg1 beta-COP 1779:1786 Silencing beta-COP 1769:1786 Silencing beta-COP 1769:1786 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 2 7 theme largest 311:317 arg1 giantin 341:347 giantin 341:347 giantin 341:347 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 7 theme largest 311:317 arg1 protein 332:338 largest Golgi matrix protein 311:338 largest Golgi matrix protein 311:338 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 1 8 theme negative 186:193 arg1 effect 195:200 a negative effect 184:200 a negative effect 184:200 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 6 9 theme α-1,2-mannosidase 992:1008 arg1 Characterization 874:889 Characterization 874:889 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) 874:1047 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 2 10 theme structure 298:306 arg1 alteration 280:289 alteration 280:289 alteration of the structure of largest Golgi matrix protein, giantin, 280:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 5 11 theme pair-fed 826:833 arg1 diets 850:854 pair-fed Lieber-DeCarli diets 826:854 pair-fed Lieber-DeCarli diets for 5 to 8 weeks 826:871 Male Wistar rats were pair-fed Lieber-DeCarli diets for 5 to 8 weeks. 27748959 9 12 theme COPI 1741:1744 arg1 vesicles 1746:1753 COPI vesicles 1741:1753 COPI vesicles at the Golgi 1741:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 5 13 theme Wistar 809:814 arg1 rats 816:819 Male Wistar rats 804:819 Male Wistar rats 804:819 Male Wistar rats were pair-fed Lieber-DeCarli diets for 5 to 8 weeks. 27748959 6 14 theme -glycoprotein 928:940 arg1 MGAT1 984:988 MGAT1 984:988 MGAT1 984:988 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 14 theme -glycoprotein 928:940 arg1 beta-1,2-N-acetylglucosaminyltransferase 942:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase 894:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1) 894:989 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 4 15 theme mouse 736:740 arg1 ADH1 742:745 mouse ADH1 736:745 mouse ADH1 (VA-13 cells) 736:759 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 4 15 theme mouse 736:740 arg1 cells 754:758 VA-13 cells 748:758 VA-13 cells 748:758 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 10 16 theme proper 1994:1999 arg1 localization 2001:2012 the proper localization 1990:2012 the proper localization of Golgi enzymes 1990:2029 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 2 17 theme apparatus 259:267 arg1 fragmentation 232:244 The fragmentation 228:244 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, 228:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 17 theme apparatus 259:267 arg1 consequence 363:373 the major consequence 353:373 the major consequence of damaging effects of EtOH-metabolism on the Golgi 353:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 8 18 theme medial-to-cis 1510:1522 arg1 relocation 1524:1533 medial-to-cis relocation 1510:1533 medial-to-cis relocation of MGAT1 and Man-II 1510:1553 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 3 19 theme EtOH 605:608 arg1 administration 610:623 EtOH administration 605:623 EtOH administration 605:623 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 6 20 theme α-mannosidase 1023:1035 arg1 Characterization 874:889 Characterization 874:889 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) 874:1047 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 2 21 theme abnormal 463:470 arg1 glycosylation 472:484 abnormal glycosylation 463:484 abnormal glycosylation 463:484 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 22 from Golgi 421:425 arg1 fragmentation 232:244 The fragmentation 228:244 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, 228:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 22 from Golgi 421:425 arg1 consequence 363:373 the major consequence 353:373 the major consequence of damaging effects of EtOH-metabolism on the Golgi 353:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 4 23 theme HepG2 707:711 arg1 cells 713:717 METHODS HepG2 cells 699:717 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) 699:759 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 10 24 theme enzymes 2023:2029 arg1 localization 2001:2012 the proper localization 1990:2012 the proper localization of Golgi enzymes 1990:2029 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 0 25 theme Alcohol-Impaired 82:97 arg1 N-Glycosylation 99:113 Alcohol-Impaired N-Glycosylation 82:113 Alcohol-Impaired N-Glycosylation 82:113 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 12 26 dep CONCLUSIONS 2178:2188 arg1 provide 2199:2205 provide 2199:2205 provide the mechanism by which EtOH-induced Golgi remodeling may significantly modify formation of N-glycans 2199:2306 CONCLUSIONS Thus, we provide the mechanism by which EtOH-induced Golgi remodeling may significantly modify formation of N-glycans. 27748959 12 27 theme Golgi 2243:2247 arg1 remodeling 2249:2258 EtOH-induced Golgi remodeling 2230:2258 EtOH-induced Golgi remodeling 2230:2258 CONCLUSIONS Thus, we provide the mechanism by which EtOH-induced Golgi remodeling may significantly modify formation of N-glycans. 27748959 4 28 theme VA-13 748:752 arg1 ADH1 742:745 mouse ADH1 736:745 mouse ADH1 (VA-13 cells) 736:759 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 4 28 theme VA-13 748:752 arg1 cells 754:758 VA-13 cells 748:758 VA-13 cells 748:758 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 2 29 theme effects 387:393 arg1 fragmentation 232:244 The fragmentation 228:244 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, 228:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 29 theme effects 387:393 arg1 consequence 363:373 the major consequence 353:373 the major consequence of damaging effects of EtOH-metabolism on the Golgi 353:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 5 30 theme Male 804:807 arg1 rats 816:819 Male Wistar rats 804:819 Male Wistar rats 804:819 Male Wistar rats were pair-fed Lieber-DeCarli diets for 5 to 8 weeks. 27748959 7 31 from loss 1241:1244 arg1 receptor 1292:1299 asialoglycoprotein receptor 1273:1299 asialoglycoprotein receptor 1273:1299 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 6 32 theme VA-13 1067:1071 arg1 cells 1073:1077 VA-13 cells 1067:1077 VA-13 cells 1067:1077 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 10 33 theme EtOH 1844:1847 arg1 effect 1849:1854 the EtOH effect 1840:1854 the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi 1840:1901 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 0 34 theme Ethanol-Induced 9:23 arg1 Disorganization 31:45 Ethanol-Induced Golgi Disorganization 9:45 Ethanol-Induced Golgi Disorganization 9:45 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 3 35 theme Golgi 644:648 arg1 enzymes 662:668 Golgi residential enzymes 644:668 Golgi residential enzymes involved in N-glycosylation 644:696 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 8 36 theme Golgi 1422:1426 arg1 disorganization 1428:1442 Golgi disorganization 1422:1442 Golgi disorganization 1422:1442 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 2 37 theme EtOH-metabolism 398:412 arg1 effects 387:393 damaging effects 378:393 damaging effects of EtOH-metabolism on the Golgi 378:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 0 38 theme Disorganization 31:45 arg1 Study 0:4 Study 0:4 Study of Ethanol-Induced Golgi Disorganization 0:45 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 12 39 theme EtOH-induced 2230:2241 arg1 remodeling 2249:2258 EtOH-induced Golgi remodeling 2230:2258 EtOH-induced Golgi remodeling 2230:2258 CONCLUSIONS Thus, we provide the mechanism by which EtOH-induced Golgi remodeling may significantly modify formation of N-glycans. 27748959 2 40 from effects 387:393 arg1 Golgi 421:425 the Golgi 417:425 the Golgi 417:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 10 41 theme COPI 2053:2056 arg1 impairment 2039:2048 impairment 2039:2048 impairment of COPI 2039:2056 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 2 42 theme protein 332:338 arg1 structure 298:306 the structure 294:306 the structure of largest Golgi matrix protein, giantin, 294:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 7 43 gly asialoglycoprotein 1273:1290 arg1 asialoglycoprotein 1273:1290 asialoglycoprotein receptor 1273:1299 asialoglycoprotein receptor 1273:1299 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 6 44 theme structured 1129:1138 arg1 microscopy 1153:1162 three-dimensional structured illumination microscopy 1111:1162 three-dimensional structured illumination microscopy (3D SIM) 1111:1171 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 44 theme structured 1129:1138 arg1 SIM 1168:1170 3D SIM 1165:1170 3D SIM 1165:1170 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 3 45 theme enzymes 662:668 arg1 function 632:639 function 632:639 function of Golgi residential enzymes involved in N-glycosylation 632:696 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 2 46 theme major 357:361 arg1 fragmentation 232:244 The fragmentation 228:244 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, 228:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 46 theme major 357:361 arg1 consequence 363:373 the major consequence 353:373 the major consequence of damaging effects of EtOH-metabolism on the Golgi 353:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 9 47 theme factor 1690:1695 arg1 GTPase 1706:1711 ADP-ribosylation factor 1 (Arf1) GTPase 1673:1711 ADP-ribosylation factor 1 (Arf1) GTPase 1673:1711 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 7 48 theme sialylated 1249:1258 arg1 N-glycans 1260:1268 sialylated N-glycans 1249:1268 sialylated N-glycans 1249:1268 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 2 49 theme Golgi 319:323 arg1 giantin 341:347 giantin 341:347 giantin 341:347 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 49 theme Golgi 319:323 arg1 protein 332:338 largest Golgi matrix protein 311:338 largest Golgi matrix protein 311:338 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 6 50 theme Golgi-associated 894:909 arg1 MGAT1 984:988 MGAT1 984:988 MGAT1 984:988 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 50 theme Golgi-associated 894:909 arg1 beta-1,2-N-acetylglucosaminyltransferase 942:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase 894:981 Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1) 894:989 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 8 51 theme Further 1358:1364 arg1 analysis 1366:1373 Further analysis 1358:1373 Further analysis by 3D SIM 1358:1383 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 10 52 theme Arf1 1822:1825 arg1 expression 1791:1800 expression 1791:1800 expression 1791:1800 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 10 52 theme Arf1 1822:1825 arg1 beta-COP 1779:1786 Silencing beta-COP 1769:1786 Silencing beta-COP 1769:1786 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 10 53 theme Arf1 1963:1966 arg1 activation 1949:1958 activation 1949:1958 activation of Arf1 1949:1966 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 3 54 theme Golgi 541:545 arg1 morphology 547:556 Golgi morphology 541:556 Golgi morphology 541:556 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 3 55 dep effect 598:603 arg1 has 625:627 has 625:627 has on function of Golgi residential enzymes involved in N-glycosylation 625:696 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 10 56 theme mutant 1815:1820 arg1 T31N 1827:1830 T31N 1827:1830 T31N 1827:1830 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 10 56 theme mutant 1815:1820 arg1 Arf1 1822:1825 GDP-bound mutant Arf1 1805:1825 GDP-bound mutant Arf1(T31N) 1805:1831 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 12 57 theme N-glycans 2298:2306 arg1 formation 2285:2293 formation 2285:2293 formation of N-glycans 2285:2306 CONCLUSIONS Thus, we provide the mechanism by which EtOH-induced Golgi remodeling may significantly modify formation of N-glycans. 27748959 4 58 theme METHODS 699:705 arg1 cells 713:717 METHODS HepG2 cells 699:717 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) 699:759 METHODS HepG2 cells transfected with mouse ADH1 (VA-13 cells) were treated with 35 mM EtOH for 72 hours. 27748959 5 59 theme Lieber-DeCarli 835:848 arg1 diets 850:854 pair-fed Lieber-DeCarli diets 826:854 pair-fed Lieber-DeCarli diets for 5 to 8 weeks 826:871 Male Wistar rats were pair-fed Lieber-DeCarli diets for 5 to 8 weeks. 27748959 6 60 theme beta-1,2-N-acetylglucosaminyltransferase 942:981 arg1 Characterization 874:889 Characterization 874:889 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) 874:1047 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 10 61 dep alters 1983:1988 arg1 ii 1974:1975 ii 1974:1975 ii 1974:1975 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 10 62 theme GDP-bound 1805:1813 arg1 T31N 1827:1830 T31N 1827:1830 T31N 1827:1830 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 10 62 theme GDP-bound 1805:1813 arg1 Arf1 1822:1825 GDP-bound mutant Arf1 1805:1825 GDP-bound mutant Arf1(T31N) 1805:1831 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 9 63 from Golgi 1762:1766 arg1 deficiency 1727:1736 a deficiency 1725:1736 a deficiency in COPI vesicles at the Golgi 1725:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 63 from Golgi 1762:1766 arg1 vesicles 1746:1753 COPI vesicles 1741:1753 COPI vesicles at the Golgi 1741:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 6 64 theme illumination 1140:1151 arg1 microscopy 1153:1162 three-dimensional structured illumination microscopy 1111:1162 three-dimensional structured illumination microscopy (3D SIM) 1111:1171 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 64 theme illumination 1140:1151 arg1 SIM 1168:1170 3D SIM 1165:1170 3D SIM 1165:1170 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 2 65 theme Golgi 253:257 arg1 apparatus 259:267 the Golgi apparatus 249:267 the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, 249:348 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 8 66 theme MGAT1 1538:1542 arg1 relocation 1524:1533 medial-to-cis relocation 1510:1533 medial-to-cis relocation of MGAT1 and Man-II 1510:1553 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 0 67 theme N-Glycosylation 99:113 arg1 Mechanism 69:77 the Potential Mechanism 55:77 the Potential Mechanism of Alcohol-Impaired N-Glycosylation 55:113 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 9 68 theme different 1562:1570 arg1 microscopy 1603:1612 electron microscopy 1594:1612 electron microscopy 1594:1612 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 68 theme different 1562:1570 arg1 approaches 1572:1581 different approaches 1562:1581 different approaches 1562:1581 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 8 69 theme Man-II 1548:1553 arg1 relocation 1524:1533 medial-to-cis relocation 1510:1533 medial-to-cis relocation of MGAT1 and Man-II 1510:1553 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 1 70 dep BACKGROUND 116:125 arg1 known 133:137 known 133:137 is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation 130:225 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 10 71 theme Golgi 2017:2021 arg1 enzymes 2023:2029 Golgi enzymes 2017:2029 Golgi enzymes 2017:2029 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 7 72 theme EtOH 1206:1209 arg1 administration 1211:1224 EtOH administration 1206:1224 EtOH administration 1206:1224 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 9 73 from deficiency 1727:1736 arg1 vesicles 1746:1753 COPI vesicles 1741:1753 COPI vesicles at the Golgi 1741:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 73 from deficiency 1727:1736 arg1 Golgi 1762:1766 the Golgi 1758:1766 the Golgi 1758:1766 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 74 theme electron 1594:1601 arg1 microscopy 1603:1612 electron microscopy 1594:1612 electron microscopy 1594:1612 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 6 75 theme rat 1083:1085 arg1 hepatocytes 1087:1097 rat hepatocytes 1083:1097 rat hepatocytes 1083:1097 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 76 dep performed 1054:1062 arg1 followed 1099:1106 followed 1099:1106 followed by three-dimensional structured illumination microscopy (3D SIM) 1099:1171 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 2 77 theme damaging 378:385 arg1 effects 387:393 damaging effects 378:393 damaging effects of EtOH-metabolism on the Golgi 378:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 0 78 theme Golgi 25:29 arg1 Disorganization 31:45 Ethanol-Induced Golgi Disorganization 9:45 Ethanol-Induced Golgi Disorganization 9:45 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 7 79 theme high-mannose-type 1315:1331 arg1 N-glycans 1333:1341 the high-mannose-type N-glycans 1311:1341 the high-mannose-type N-glycans 1311:1341 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 10 80 dep blocks 1942:1947 arg1 i 1921:1921 i 1921:1921 i 1921:1921 Silencing beta-COP or expression of GDP-bound mutant Arf1(T31N) mimics the EtOH effect on retaining MGAT1 and Man-II at the cis-Golgi, suggesting that (i) EtOH specifically blocks activation of Arf1, and (ii) EtOH alters the proper localization of Golgi enzymes through impairment of COPI. 27748959 8 81 theme EtOH 1399:1402 arg1 treatment 1404:1412 EtOH treatment 1399:1412 EtOH treatment 1399:1412 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 6 82 theme three-dimensional 1111:1127 arg1 microscopy 1153:1162 three-dimensional structured illumination microscopy 1111:1162 three-dimensional structured illumination microscopy (3D SIM) 1111:1171 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 6 82 theme three-dimensional 1111:1127 arg1 SIM 1168:1170 3D SIM 1165:1170 3D SIM 1165:1170 Characterization of Golgi-associated mannosyl (α-1,3-)-glycoprotein beta-1,2-N-acetylglucosaminyltransferase (MGAT1), α-1,2-mannosidase (Man-I), and α-mannosidase II (Man-II) were performed in VA-13 cells and rat hepatocytes followed by three-dimensional structured illumination microscopy (3D SIM). 27748959 7 83 gly sialylated 1249:1258 arg1 N-glycans 1260:1268 sialylated N-glycans 1249:1268 sialylated N-glycans 1249:1268 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 1 84 theme protein 205:211 arg1 glycosylation 213:225 protein glycosylation 205:225 protein glycosylation 205:225 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 9 85 theme EtOH 1632:1635 arg1 treatment 1637:1645 EtOH treatment 1632:1645 EtOH treatment 1632:1645 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 1 86 contain have 179:182 arg1 ethanol 144:150 ethanol 144:150 ethanol (EtOH) 144:157 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 1 86 contain have 179:182 arg1 metabolites 167:177 its metabolites 163:177 its metabolites 163:177 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 1 86 contain have 179:182 arg2 effect 195:200 a negative effect 184:200 a negative effect 184:200 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 1 86 contain have 179:182 arg1 EtOH 153:156 EtOH 153:156 EtOH 153:156 BACKGROUND It is known that ethanol (EtOH) and its metabolites have a negative effect on protein glycosylation. 27748959 0 87 theme Potential 59:67 arg1 Mechanism 69:77 the Potential Mechanism 55:77 the Potential Mechanism of Alcohol-Impaired N-Glycosylation 55:113 Study of Ethanol-Induced Golgi Disorganization Reveals the Potential Mechanism of Alcohol-Impaired N-Glycosylation. 27748959 2 88 from consequence 363:373 arg1 Golgi 421:425 the Golgi 417:425 the Golgi 417:425 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 7 89 theme asialoglycoprotein 1273:1290 arg1 receptor 1292:1299 asialoglycoprotein receptor 1273:1299 asialoglycoprotein receptor 1273:1299 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 8 90 theme 3D 1378:1379 arg1 SIM 1381:1383 3D SIM 1378:1383 3D SIM 1378:1383 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 27748959 3 91 theme residential 650:660 arg1 enzymes 662:668 Golgi residential enzymes 644:668 Golgi residential enzymes involved in N-glycosylation 644:696 Because previously we have shown that Golgi morphology dictates glycosylation, we examined the effect EtOH administration has on function of Golgi residential enzymes involved in N-glycosylation. 27748959 2 92 theme matrix 325:330 arg1 giantin 341:347 giantin 341:347 giantin 341:347 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 2 92 theme matrix 325:330 arg1 protein 332:338 largest Golgi matrix protein 311:338 largest Golgi matrix protein 311:338 The fragmentation of the Golgi apparatus induced by alteration of the structure of largest Golgi matrix protein, giantin, is the major consequence of damaging effects of EtOH-metabolism on the Golgi; however, the link between this and abnormal glycosylation remains unknown. 27748959 9 93 theme ADP-ribosylation 1673:1688 arg1 Arf1 1700:1703 Arf1 1700:1703 Arf1 1700:1703 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 9 93 theme ADP-ribosylation 1673:1688 arg1 factor 1690:1695 ADP-ribosylation factor 1 1673:1697 ADP-ribosylation factor 1 (Arf1) GTPase 1673:1711 Using different approaches, including electron microscopy, we revealed that EtOH treatment results in dysfunction of ADP-ribosylation factor 1 (Arf1) GTPase followed by a deficiency in COPI vesicles at the Golgi. 27748959 7 94 theme N-glycans 1260:1268 arg1 loss 1241:1244 the loss 1237:1244 the loss of sialylated N-glycans on asialoglycoprotein receptor 1237:1299 RESULTS First, we detected that EtOH administration results in the loss of sialylated N-glycans on asialoglycoprotein receptor; however, the high-mannose-type N-glycans are increased. 27748959 8 95 theme cis-Golgi 1460:1468 arg1 localization 1470:1481 cis-Golgi localization 1460:1481 cis-Golgi localization for Man-I 1460:1491 Further analysis by 3D SIM revealed that EtOH treatment despite Golgi disorganization does not change cis-Golgi localization for Man-I, but does induce medial-to-cis relocation of MGAT1 and Man-II. 24286971 0 0 theme oligosaccharide 80:94 arg1 profile 149:155 the N-glycosylation profile 129:155 the N-glycosylation profile of a monoclonal antibody 129:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 0 theme oligosaccharide 80:94 arg1 distribution 96:107 the intracellular lipid-linked oligosaccharide distribution 49:107 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 0 theme oligosaccharide 80:94 arg1 occupancy 115:123 site occupancy 110:123 site occupancy 110:123 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 2 1 theme non-glycosylated 418:433 arg1 Mab 435:437 non-glycosylated Mab 418:437 non-glycosylated Mab 418:437 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 4 2 theme LLO 786:788 arg1 LLO 786:788 full length LLO 774:788 full length LLO (Glc3Man9GlcNac2) 774:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 2 theme LLO 786:788 arg1 amount 764:769 the amount 760:769 the amount of full length LLO (Glc3Man9GlcNac2) 760:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 2 theme LLO 786:788 arg1 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 3 link lipid-linked 661:672 arg1 oligosaccharides 674:689 intracellular lipid-linked oligosaccharides 647:689 intracellular lipid-linked oligosaccharides (LLOs) 647:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 3 link lipid-linked 661:672 arg1 LLOs 692:695 LLOs 692:695 LLOs 692:695 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 0 4 theme lipid-linked 67:78 arg1 oligosaccharide 80:94 lipid-linked oligosaccharide 67:94 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 4 5 theme length 779:784 arg1 LLO 786:788 full length LLO 774:788 full length LLO (Glc3Man9GlcNac2) 774:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 5 theme length 779:784 arg1 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 8 6 theme chimeric 1346:1353 arg1 EG2 1365:1367 EG2 1365:1367 EG2 1365:1367 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 8 6 theme chimeric 1346:1353 arg1 antibody 1355:1362 chimeric antibody 1346:1362 the chimeric antibody (EG2) 1342:1368 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 4 7 from reduction 747:755 arg1 LLO 786:788 full length LLO 774:788 full length LLO (Glc3Man9GlcNac2) 774:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 7 from reduction 747:755 arg1 amount 764:769 the amount 760:769 the amount of full length LLO (Glc3Man9GlcNac2) 760:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 7 from reduction 747:755 arg1 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 1 8 theme cultures 328:335 arg1 0-25mM 317:322 0-25mM 317:322 0-25mM 317:322 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 8 theme cultures 328:335 arg1 concentration 302:314 the glucose concentration 290:314 the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h 290:387 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 6 9 with supplementation 1094:1108 arg1 cocktail 1117:1124 a cocktail 1115:1124 a cocktail of uridine, manganese and galactose 1115:1160 The GI increased to 0.83 following media supplementation with a cocktail of uridine, manganese and galactose. 24286971 2 10 gly non-glycosylated 418:433 arg1 Mab 435:437 non-glycosylated Mab 418:437 non-glycosylated Mab 418:437 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 4 11 theme smaller 844:850 arg1 Man2-5GlcNAc2 870:882 Man2-5GlcNAc2 870:882 Man2-5GlcNAc2 870:882 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 11 theme smaller 844:850 arg1 mannosyl-glycans 852:867 the smaller mannosyl-glycans 840:867 the smaller mannosyl-glycans (Man2-5GlcNAc2) 840:883 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 8 12 theme molecular 1392:1400 arg1 weight 1402:1407 its low molecular weight 1384:1407 its low molecular weight 1384:1407 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 1 13 gly glycosylation 187:199 arg1 EG2 245:247 EG2 245:247 EG2 245:247 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 13 gly glycosylation 187:199 arg1 antibody 235:242 a chimeric heavy chain antibody 212:242 a chimeric heavy chain antibody (EG2) produced from CHO cells 212:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 6 14 theme media 1088:1092 arg1 supplementation 1094:1108 media supplementation 1088:1108 media supplementation with a cocktail of uridine, manganese and galactose 1088:1160 The GI increased to 0.83 following media supplementation with a cocktail of uridine, manganese and galactose. 24286971 1 15 theme CHO 264:266 arg1 cells 268:272 CHO cells 264:272 CHO cells 264:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 8 16 theme low 1388:1390 arg1 weight 1402:1407 its low molecular weight 1384:1407 its low molecular weight 1384:1407 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 9 17 theme Mab 1535:1537 arg1 production 1539:1548 Mab production 1535:1548 Mab production 1535:1548 These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. 24286971 5 18 theme Glycan 886:891 arg1 microheterogeneity 893:910 Glycan microheterogeneity 886:910 Glycan microheterogeneity 886:910 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 19 dep indices 962:968 arg1 SI 978:979 SI 978:979 SI 978:979 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 19 dep indices 962:968 arg1 GI 971:972 GI 971:972 GI 971:972 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 19 dep indices 962:968 arg1 indices 962:968 galactosylation and sialylation indices 930:968 galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake 930:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 0 20 theme site 110:113 arg1 distribution 96:107 the intracellular lipid-linked oligosaccharide distribution 49:107 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 20 theme site 110:113 arg1 occupancy 115:123 site occupancy 110:123 site occupancy 110:123 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 1 21 theme high 352:355 arg1 density 357:363 high density 352:363 high density (>10(6)ml(-1)) over 24h 352:387 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 3 22 gly non-glycosylated 581:596 arg1 fraction 602:609 a 45% non-glycosylated Mab fraction 575:609 a 45% non-glycosylated Mab fraction 575:609 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 7 23 dep higher 1185:1190 arg1 DP12 1229:1232 DP12 1229:1232 DP12 1229:1232 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 7 23 dep higher 1185:1190 arg1 antibody 1219:1226 a fully humanized antibody 1201:1226 a fully humanized antibody (DP12) produced under the similar conditions 1201:1271 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 7 23 dep higher 1185:1190 arg1 antibodies 1288:1297 similar antibodies 1280:1297 similar antibodies reported in the literature 1280:1324 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 5 24 theme cell 1023:1026 arg1 uptake 1045:1050 the cell specific glucose uptake 1019:1050 the cell specific glucose uptake 1019:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 4 25 theme full 774:777 arg1 LLO 786:788 full length LLO 774:788 full length LLO (Glc3Man9GlcNac2) 774:806 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 25 theme full 774:777 arg1 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Glc3Man9GlcNac2 791:805 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 26 theme lipid-linked 661:672 arg1 oligosaccharides 674:689 intracellular lipid-linked oligosaccharides 647:689 intracellular lipid-linked oligosaccharides (LLOs) 647:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 26 theme lipid-linked 661:672 arg1 LLOs 692:695 LLOs 692:695 LLOs 692:695 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 1 27 theme glycosylation 187:199 arg1 pattern 201:207 The glycosylation pattern 183:207 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells 183:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 6 28 theme galactose 1152:1160 arg1 cocktail 1117:1124 a cocktail 1115:1124 a cocktail of uridine, manganese and galactose 1115:1160 The GI increased to 0.83 following media supplementation with a cocktail of uridine, manganese and galactose. 24286971 4 29 theme intracellular 647:659 arg1 oligosaccharides 674:689 intracellular lipid-linked oligosaccharides 647:689 intracellular lipid-linked oligosaccharides (LLOs) 647:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 29 theme intracellular 647:659 arg1 LLOs 692:695 LLOs 692:695 LLOs 692:695 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 9 30 theme fed-batch 1512:1520 arg1 cultures 1522:1529 fed-batch cultures 1512:1529 fed-batch cultures for Mab production 1512:1548 These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. 24286971 0 31 theme glucose 20:26 arg1 availability 4:15 The availability 0:15 The availability of glucose to CHO cells 0:39 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 8 32 theme unusual 1413:1419 arg1 structure 1421:1429 unusual structure 1413:1429 unusual structure 1413:1429 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 2 33 theme cells 487:491 arg1 time 479:482 the exposure time 466:482 the exposure time of cells to media depleted of glucose 466:520 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 9 34 theme low 1480:1482 arg1 substrate 1484:1492 the low substrate 1476:1492 the low substrate that may occur in fed-batch cultures for Mab production 1476:1548 These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. 24286971 7 35 theme humanized 1209:1217 arg1 DP12 1229:1232 DP12 1229:1232 DP12 1229:1232 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 7 35 theme humanized 1209:1217 arg1 antibody 1219:1226 a fully humanized antibody 1201:1226 a fully humanized antibody (DP12) produced under the similar conditions 1201:1271 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 5 36 theme glucose 1037:1043 arg1 uptake 1045:1050 the cell specific glucose uptake 1019:1050 the cell specific glucose uptake 1019:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 4 37 theme concomitant 816:826 arg1 increase 828:835 a concomitant increase 814:835 a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2) 814:883 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 0 38 gly occupancy 115:123 arg2 antibody 173:180 a monoclonal antibody 160:180 a monoclonal antibody 160:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 38 gly occupancy 115:123 arg2 site 110:113 site occupancy 110:123 site occupancy 110:123 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 38 gly occupancy 115:123 arg1 antibody 173:180 a monoclonal antibody 160:180 a monoclonal antibody 160:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 9 39 from relation 1464:1471 arg1 important 1451:1459 important 1451:1459 important 1451:1459 These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. 24286971 8 40 theme antibody 1355:1362 arg1 due 1377:1379 due 1377:1379 due 1377:1379 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 8 40 theme antibody 1355:1362 arg1 GI 1336:1337 The high GI 1327:1337 The high GI of the chimeric antibody (EG2) 1327:1368 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 0 41 theme CHO 31:33 arg1 cells 35:39 CHO cells 31:39 CHO cells 31:39 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 7 42 theme similar 1280:1286 arg1 antibodies 1288:1297 similar antibodies 1280:1297 similar antibodies reported in the literature 1280:1324 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 1 43 theme chimeric 214:221 arg1 EG2 245:247 EG2 245:247 EG2 245:247 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 43 theme chimeric 214:221 arg1 antibody 235:242 a chimeric heavy chain antibody 212:242 a chimeric heavy chain antibody (EG2) produced from CHO cells 212:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 2 44 theme resulting 394:402 arg1 proportion 404:413 The resulting proportion 390:413 The resulting proportion of non-glycosylated Mab 390:437 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 0 45 theme N-glycosylation 133:147 arg1 profile 149:155 the N-glycosylation profile 129:155 the N-glycosylation profile of a monoclonal antibody 129:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 45 theme N-glycosylation 133:147 arg1 distribution 96:107 the intracellular lipid-linked oligosaccharide distribution 49:107 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 4 46 from increase 828:835 arg1 Man2-5GlcNAc2 870:882 Man2-5GlcNAc2 870:882 Man2-5GlcNAc2 870:882 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 46 from increase 828:835 arg1 mannosyl-glycans 852:867 the smaller mannosyl-glycans 840:867 the smaller mannosyl-glycans (Man2-5GlcNAc2) 840:883 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 5 47 theme sialylation 950:960 arg1 SI 978:979 SI 978:979 SI 978:979 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 47 theme sialylation 950:960 arg1 GI 971:972 GI 971:972 GI 971:972 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 47 theme sialylation 950:960 arg1 indices 962:968 galactosylation and sialylation indices 930:968 galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake 930:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 1 48 theme heavy 223:227 arg1 EG2 245:247 EG2 245:247 EG2 245:247 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 48 theme heavy 223:227 arg1 antibody 235:242 a chimeric heavy chain antibody 212:242 a chimeric heavy chain antibody (EG2) produced from CHO cells 212:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 49 theme glucose 294:300 arg1 0-25mM 317:322 0-25mM 317:322 0-25mM 317:322 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 49 theme glucose 294:300 arg1 concentration 302:314 the glucose concentration 290:314 the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h 290:387 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 7 50 theme similar 1254:1260 arg1 conditions 1262:1271 the similar conditions 1250:1271 the similar conditions 1250:1271 This is significantly higher than for a fully humanized antibody (DP12) produced under the similar conditions or for similar antibodies reported in the literature. 24286971 5 51 theme direct 997:1002 arg1 correlation 1004:1014 a direct correlation 995:1014 a direct correlation to the cell specific glucose uptake 995:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 0 52 link lipid-linked 67:78 arg1 oligosaccharide 80:94 lipid-linked oligosaccharide 67:94 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 4 53 theme state 631:635 arg1 levels 637:642 steady state levels 624:642 steady state levels of intracellular lipid-linked oligosaccharides (LLOs) 624:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 2 54 theme exposure 470:477 arg1 time 479:482 the exposure time 466:482 the exposure time of cells to media depleted of glucose 466:520 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 3 55 theme non-glycosylated 581:596 arg1 fraction 602:609 a 45% non-glycosylated Mab fraction 575:609 a 45% non-glycosylated Mab fraction 575:609 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 6 56 theme uridine 1129:1135 arg1 cocktail 1117:1124 a cocktail 1115:1124 a cocktail of uridine, manganese and galactose 1115:1160 The GI increased to 0.83 following media supplementation with a cocktail of uridine, manganese and galactose. 24286971 4 57 theme steady 624:629 arg1 levels 637:642 steady state levels 624:642 steady state levels of intracellular lipid-linked oligosaccharides (LLOs) 624:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 9 58 from important 1451:1459 arg1 relation 1464:1471 relation 1464:1471 relation to the low substrate that may occur in fed-batch cultures for Mab production 1464:1548 These findings are important in relation to the low substrate that may occur in fed-batch cultures for Mab production. 24286971 8 59 theme high 1331:1334 arg1 due 1377:1379 due 1377:1379 due 1377:1379 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 8 59 theme high 1331:1334 arg1 GI 1336:1337 The high GI 1327:1337 The high GI of the chimeric antibody (EG2) 1327:1368 The high GI of the chimeric antibody (EG2) may be due to its low molecular weight and unusual structure. 24286971 0 60 theme antibody 173:180 arg1 profile 149:155 the N-glycosylation profile 129:155 the N-glycosylation profile of a monoclonal antibody 129:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 60 theme antibody 173:180 arg1 distribution 96:107 the intracellular lipid-linked oligosaccharide distribution 49:107 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 60 theme antibody 173:180 arg1 occupancy 115:123 site occupancy 110:123 site occupancy 110:123 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 3 61 theme Mab 598:600 arg1 fraction 602:609 a 45% non-glycosylated Mab fraction 575:609 a 45% non-glycosylated Mab fraction 575:609 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 4 62 theme glucose 716:722 arg1 limitation 724:733 glucose limitation 716:733 glucose limitation 716:733 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 6 63 theme manganese 1138:1146 arg1 cocktail 1117:1124 a cocktail 1115:1124 a cocktail of uridine, manganese and galactose 1115:1160 The GI increased to 0.83 following media supplementation with a cocktail of uridine, manganese and galactose. 24286971 0 64 gly N-glycosylation 133:147 arg1 antibody 173:180 a monoclonal antibody 160:180 a monoclonal antibody 160:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 1 65 theme chain 229:233 arg1 EG2 245:247 EG2 245:247 EG2 245:247 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 65 theme chain 229:233 arg1 antibody 235:242 a chimeric heavy chain antibody 212:242 a chimeric heavy chain antibody (EG2) produced from CHO cells 212:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 0 66 theme monoclonal 162:171 arg1 antibody 173:180 a monoclonal antibody 160:180 a monoclonal antibody 160:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 3 67 theme 45 577:578 arg1 % 579:579 % 579:579 % 579:579 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 3 68 theme full 554:557 arg1 24h 559:561 the full 24h 550:561 the full 24h 550:561 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 4 69 theme oligosaccharides 674:689 arg1 levels 637:642 steady state levels 624:642 steady state levels of intracellular lipid-linked oligosaccharides (LLOs) 624:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 4 70 theme levels 637:642 arg1 Analysis 612:619 Analysis 612:619 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) 612:696 Analysis of steady state levels of intracellular lipid-linked oligosaccharides (LLOs) showed that under glucose limitation there was a reduction in the amount of full length LLO (Glc3Man9GlcNac2), with a concomitant increase in the smaller mannosyl-glycans (Man2-5GlcNAc2). 24286971 1 71 theme antibody 235:242 arg1 pattern 201:207 The glycosylation pattern 183:207 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells 183:272 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 0 72 theme intracellular 53:65 arg1 profile 149:155 the N-glycosylation profile 129:155 the N-glycosylation profile of a monoclonal antibody 129:180 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 72 theme intracellular 53:65 arg1 distribution 96:107 the intracellular lipid-linked oligosaccharide distribution 49:107 the intracellular lipid-linked oligosaccharide distribution 49:107 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 0 72 theme intracellular 53:65 arg1 occupancy 115:123 site occupancy 110:123 site occupancy 110:123 The availability of glucose to CHO cells affects the intracellular lipid-linked oligosaccharide distribution, site occupancy and the N-glycosylation profile of a monoclonal antibody. 24286971 3 73 theme % 579:579 arg1 fraction 602:609 a 45% non-glycosylated Mab fraction 575:609 a 45% non-glycosylated Mab fraction 575:609 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 3 74 theme glucose 538:544 arg1 Deprivation 523:533 Deprivation 523:533 Deprivation of glucose for the full 24h 523:561 Deprivation of glucose for the full 24h resulted in a 45% non-glycosylated Mab fraction. 24286971 2 75 theme Mab 435:437 arg1 proportion 404:413 The resulting proportion 390:413 The resulting proportion of non-glycosylated Mab 390:437 The resulting proportion of non-glycosylated Mab was directly correlated to the exposure time of cells to media depleted of glucose. 24286971 5 76 theme specific 1028:1035 arg1 uptake 1045:1050 the cell specific glucose uptake 1019:1050 the cell specific glucose uptake 1019:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 77 theme galactosylation 930:944 arg1 SI 978:979 SI 978:979 SI 978:979 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 77 theme galactosylation 930:944 arg1 GI 971:972 GI 971:972 GI 971:972 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 5 77 theme galactosylation 930:944 arg1 indices 962:968 galactosylation and sialylation indices 930:968 galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake 930:1050 Glycan microheterogeneity was quantified by galactosylation and sialylation indices (GI and SI) which showed a direct correlation to the cell specific glucose uptake. 24286971 1 78 dep density 357:363 arg1 ml 372:373 >10(6)ml 366:373 >10(6)ml(-1) 366:377 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 24286971 1 78 dep density 357:363 arg1 -1 375:376 -1 375:376 -1 375:376 The glycosylation pattern of a chimeric heavy chain antibody (EG2) produced from CHO cells was affected by the glucose concentration (0-25mM) of cultures established at high density (>10(6)ml(-1)) over 24h. 25545631 12 0 theme metabolic 1824:1832 arg1 distribution 1839:1850 the metabolic flux distribution 1820:1850 the metabolic flux distribution 1820:1850 Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. 25545631 5 1 theme cell 681:684 arg1 viability 686:694 higher cell viability 674:694 higher cell viability 674:694 Experimental results showed higher cell viability with decreased metabolic rates. 25545631 7 2 theme exometabolite 1094:1106 arg1 data 1108:1111 exometabolite data 1094:1111 exometabolite data from stationary phase of cultures with or without a temperature shift 1094:1181 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 6 3 theme decreased 890:898 arg1 proportion 900:909 a decreased proportion 888:909 a decreased proportion of the more processed glycan structures on the mAb constant region 888:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 8 4 theme carbon 1231:1236 arg1 species 1238:1244 carbon species 1231:1244 carbon species 1231:1244 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 10 5 theme NSD 1554:1556 arg1 availability 1558:1569 NSD availability 1554:1569 NSD availability 1554:1569 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 2 6 theme cellular 275:282 arg1 processes 284:292 dynamic cellular processes 267:292 dynamic cellular processes 267:292 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 1 7 theme protein 192:198 arg1 production 200:209 recombinant protein production 180:209 recombinant protein production 180:209 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 8 8 theme Estimated 1184:1192 arg1 fluxomes 1194:1201 Estimated fluxomes 1184:1201 Estimated fluxomes 1184:1201 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 9 9 link N-linked 1395:1402 arg1 glycan 1404:1409 N-linked glycan branching and elongation 1395:1434 glycan 1404:1409 Expression of the glycosyltransferases that are responsible for N-linked glycan branching and elongation were significantly lower at 32°C. 25545631 4 10 theme stationary 628:637 arg1 phase 639:643 late exponential/early stationary phase 605:643 late exponential/early stationary phase 605:643 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 6 11 theme mAb 958:960 arg1 region 971:976 the mAb constant region 954:976 the mAb constant region 954:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 4 12 theme hamster 472:478 arg1 cells 492:496 Chinese hamster ovary (CHO) cells 464:496 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) 464:535 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 8 13 theme more 1287:1290 arg1 energy 1292:1297 more energy 1287:1297 more energy 1287:1297 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 7 14 theme cell 1004:1007 arg1 metabolism 1009:1018 CHO cell metabolism 1000:1018 CHO cell metabolism 1000:1018 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 6 15 theme glycan 933:938 arg1 structures 940:949 the more processed glycan structures 914:949 the more processed glycan structures on the mAb constant region 914:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 7 16 theme flux 1029:1032 arg1 FBA 1052:1054 FBA 1052:1054 FBA 1052:1054 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 7 16 theme flux 1029:1032 arg1 analysis 1042:1049 flux balance analysis 1029:1049 flux balance analysis (FBA) 1029:1055 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 12 17 theme feeding 1754:1760 arg1 strategies 1762:1771 Better feeding strategies 1747:1771 Better feeding strategies 1747:1771 Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. 25545631 1 18 theme industrial 144:153 arg1 application 72:82 The application 68:82 The application of mild hypothermic conditions to cell culture 68:129 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 1 18 theme industrial 144:153 arg1 practice 155:162 a routine industrial practice 134:162 a routine industrial practice used to improve recombinant protein production 134:209 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 7 19 from phase 1129:1133 arg1 data 1108:1111 exometabolite data 1094:1111 exometabolite data from stationary phase of cultures with or without a temperature shift 1094:1181 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 5 20 theme decreased 701:709 arg1 rates 721:725 decreased metabolic rates 701:725 decreased metabolic rates 701:725 Experimental results showed higher cell viability with decreased metabolic rates. 25545631 5 21 theme higher 674:679 arg1 viability 686:694 higher cell viability 674:694 higher cell viability 674:694 Experimental results showed higher cell viability with decreased metabolic rates. 25545631 4 22 theme monoclonal 511:520 arg1 mAb 532:534 mAb 532:534 mAb 532:534 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 4 22 theme monoclonal 511:520 arg1 antibody 522:529 a monoclonal antibody 509:529 a monoclonal antibody (mAb) 509:535 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 3 23 theme protein 441:447 arg1 glycosylation 449:461 protein glycosylation 441:461 protein glycosylation 441:461 In this study, we investigated the impact of mild hypothermia on protein glycosylation. 25545631 2 24 dep bioprocess 340:349 arg1 design 351:356 design 351:356 design 351:356 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 3 25 theme hypothermia 426:436 arg1 impact 411:416 the impact 407:416 the impact of mild hypothermia on protein glycosylation 407:461 In this study, we investigated the impact of mild hypothermia on protein glycosylation. 25545631 1 26 theme hypothermic 92:102 arg1 conditions 104:113 mild hypothermic conditions 87:113 mild hypothermic conditions 87:113 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 6 27 from reduction 813:821 arg1 proportion 900:909 a decreased proportion 888:909 a decreased proportion of the more processed glycan structures on the mAb constant region 888:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 27 from reduction 813:821 arg1 concentrations 869:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 2 28 theme regulation 253:262 arg1 understanding 232:244 a thorough understanding 221:244 a thorough understanding of the regulation of dynamic cellular processes at lower temperatures 221:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 2 28 theme regulation 253:262 arg1 necessary 319:327 necessary 319:327 necessary 319:327 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 10 29 theme mAb 1503:1505 arg1 result 1475:1480 a result 1473:1480 a result of mild hypothermia 1473:1500 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 10 29 theme mAb 1503:1505 arg1 glycosylation 1507:1519 mAb glycosylation 1503:1519 mAb glycosylation 1503:1519 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 0 30 theme mild 9:12 arg1 hypothermia 14:24 mild hypothermia 9:24 mild hypothermia 9:24 How does mild hypothermia affect monoclonal antibody glycosylation? 25545631 10 31 theme hypothermia 1490:1500 arg1 result 1475:1480 a result 1473:1480 a result of mild hypothermia 1473:1500 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 10 31 theme hypothermia 1490:1500 arg1 glycosylation 1507:1519 mAb glycosylation 1503:1519 mAb glycosylation 1503:1519 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 11 32 theme culture 1726:1732 arg1 temperature 1734:1744 culture temperature 1726:1744 culture temperature 1726:1744 The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. 25545631 7 33 theme cultures 1138:1145 arg1 phase 1129:1133 stationary phase 1118:1133 stationary phase of cultures with or without a temperature shift 1118:1181 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 9 34 theme glycosyltransferases 1349:1368 arg1 Expression 1331:1340 Expression 1331:1340 Expression of the glycosyltransferases that are responsible for N-linked glycan branching and elongation 1331:1434 Expression of the glycosyltransferases that are responsible for N-linked glycan branching and elongation were significantly lower at 32°C. 25545631 6 35 from structures 940:949 arg1 region 971:976 the mAb constant region 954:976 the mAb constant region 954:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 11 36 theme combined 1611:1618 arg1 approach 1637:1644 The combined experimental/FBA approach 1607:1644 The combined experimental/FBA approach 1607:1644 The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. 25545631 6 37 from region 971:976 arg1 proportion 900:909 a decreased proportion 888:909 a decreased proportion of the more processed glycan structures on the mAb constant region 888:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 37 from region 971:976 arg1 concentrations 869:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 38 theme donor 857:861 arg1 concentrations 869:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 39 theme nucleotide 840:849 arg1 NSD 864:866 NSD 864:866 NSD 864:866 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 39 theme nucleotide 840:849 arg1 donor 857:861 nucleotide sugar donor 840:861 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 40 theme processed 923:931 arg1 structures 940:949 the more processed glycan structures 914:949 the more processed glycan structures on the mAb constant region 914:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 12 41 theme flux 1834:1837 arg1 distribution 1839:1850 the metabolic flux distribution 1820:1850 the metabolic flux distribution 1820:1850 Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. 25545631 11 42 theme product 1674:1680 arg1 glycosylation 1682:1694 product glycosylation 1674:1694 product glycosylation 1674:1694 The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. 25545631 5 43 theme Experimental 646:657 arg1 results 659:665 Experimental results 646:665 Experimental results 646:665 Experimental results showed higher cell viability with decreased metabolic rates. 25545631 6 44 theme specific 732:739 arg1 productivity 750:761 The specific antibody productivity 728:761 The specific antibody productivity 728:761 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 10 45 theme glycosyltransferase 1575:1593 arg1 expression 1595:1604 glycosyltransferase expression 1575:1604 glycosyltransferase expression 1575:1604 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 2 46 theme lower 297:301 arg1 temperatures 303:314 lower temperatures 297:314 lower temperatures 297:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 4 47 theme temperature 572:582 arg1 shift 584:588 a temperature shift 570:588 a temperature shift to 32°C 570:596 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 6 48 from proportion 900:909 arg1 region 971:976 the mAb constant region 954:976 the mAb constant region 954:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 2 49 theme processes 284:292 arg1 regulation 253:262 the regulation 249:262 the regulation of dynamic cellular processes at lower temperatures 249:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 0 50 theme antibody 44:51 arg1 glycosylation 53:65 monoclonal antibody glycosylation 33:65 monoclonal antibody glycosylation 33:65 How does mild hypothermia affect monoclonal antibody glycosylation? 25545631 1 51 theme recombinant 180:190 arg1 production 200:209 recombinant protein production 180:209 recombinant protein production 180:209 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 8 52 theme species 1238:1244 arg1 fluxes 1221:1226 reduced fluxes 1213:1226 reduced fluxes of carbon species towards nucleotide and NSD synthesis 1213:1281 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 6 53 from concentrations 869:882 arg1 region 971:976 the mAb constant region 954:976 the mAb constant region 954:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 5 54 theme metabolic 711:719 arg1 rates 721:725 decreased metabolic rates 701:725 decreased metabolic rates 701:725 Experimental results showed higher cell viability with decreased metabolic rates. 25545631 6 55 theme constant 962:969 arg1 region 971:976 the mAb constant region 954:976 the mAb constant region 954:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 9 56 dep glycan 1404:1409 arg1 branching 1411:1419 branching 1411:1419 branching 1411:1419 Expression of the glycosyltransferases that are responsible for N-linked glycan branching and elongation were significantly lower at 32°C. 25545631 4 57 theme exponential/early 610:626 arg1 phase 639:643 late exponential/early stationary phase 605:643 late exponential/early stationary phase 605:643 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 4 58 theme ovary 480:484 arg1 cells 492:496 Chinese hamster ovary (CHO) cells 464:496 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) 464:535 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 8 59 theme NSD 1269:1271 arg1 synthesis 1273:1281 NSD synthesis 1269:1281 NSD synthesis 1269:1281 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 7 60 theme balance 1034:1040 arg1 FBA 1052:1054 FBA 1052:1054 FBA 1052:1054 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 7 60 theme balance 1034:1040 arg1 analysis 1042:1049 flux balance analysis 1029:1049 flux balance analysis (FBA) 1029:1055 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 2 61 theme thorough 223:230 arg1 understanding 232:244 a thorough understanding 221:244 a thorough understanding of the regulation of dynamic cellular processes at lower temperatures 221:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 2 61 theme thorough 223:230 arg1 necessary 319:327 necessary 319:327 necessary 319:327 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 1 62 theme cell 118:121 arg1 culture 123:129 cell culture 118:129 cell culture 118:129 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 8 63 theme reduced 1213:1219 arg1 fluxes 1221:1226 reduced fluxes 1213:1226 reduced fluxes of carbon species towards nucleotide and NSD synthesis 1213:1281 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 6 64 theme structures 940:949 arg1 proportion 900:909 a decreased proportion 888:909 a decreased proportion of the more processed glycan structures on the mAb constant region 888:976 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 64 theme structures 940:949 arg1 concentrations 869:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 2 65 theme dynamic 267:273 arg1 processes 284:292 dynamic cellular processes 267:292 dynamic cellular processes 267:292 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 8 66 used used 1303:1306 arg2 energy 1292:1297 more energy 1287:1297 more energy 1287:1297 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 7 67 theme CHO 1000:1002 arg1 metabolism 1009:1018 CHO cell metabolism 1000:1018 CHO cell metabolism 1000:1018 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 1 68 theme routine 136:142 arg1 application 72:82 The application 68:82 The application of mild hypothermic conditions to cell culture 68:129 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 1 68 theme routine 136:142 arg1 practice 155:162 a routine industrial practice 134:162 a routine industrial practice used to improve recombinant protein production 134:209 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 12 69 theme Better 1747:1752 arg1 strategies 1762:1771 Better feeding strategies 1747:1771 Better feeding strategies 1747:1771 Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. 25545631 3 70 from impact 411:416 arg1 glycosylation 449:461 protein glycosylation 441:461 protein glycosylation 441:461 In this study, we investigated the impact of mild hypothermia on protein glycosylation. 25545631 2 71 from temperatures 303:314 arg1 understanding 232:244 a thorough understanding 221:244 a thorough understanding of the regulation of dynamic cellular processes at lower temperatures 221:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 2 71 from temperatures 303:314 arg1 regulation 253:262 the regulation 249:262 the regulation of dynamic cellular processes at lower temperatures 249:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 2 71 from temperatures 303:314 arg1 necessary 319:327 necessary 319:327 necessary 319:327 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 8 72 theme product 1312:1318 arg1 formation 1320:1328 product formation 1312:1328 product formation 1312:1328 Estimated fluxomes suggested reduced fluxes of carbon species towards nucleotide and NSD synthesis and more energy was used for product formation. 25545631 4 73 theme Chinese 464:470 arg1 cells 492:496 Chinese hamster ovary (CHO) cells 464:496 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) 464:535 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 11 74 from changes 1715:1721 arg1 temperature 1734:1744 culture temperature 1726:1744 culture temperature 1726:1744 The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. 25545631 4 75 theme CHO 487:489 arg1 cells 492:496 Chinese hamster ovary (CHO) cells 464:496 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) 464:535 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 7 76 theme temperature 1165:1175 arg1 shift 1177:1181 a temperature shift 1163:1181 a temperature shift 1163:1181 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 1 77 theme mild 87:90 arg1 conditions 104:113 mild hypothermic conditions 87:113 mild hypothermic conditions 87:113 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 2 78 from understanding 232:244 arg1 temperatures 303:314 lower temperatures 297:314 lower temperatures 297:314 However, a thorough understanding of the regulation of dynamic cellular processes at lower temperatures is necessary to enhance bioprocess design and optimization. 25545631 1 79 theme conditions 104:113 arg1 application 72:82 The application 68:82 The application of mild hypothermic conditions to cell culture 68:129 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 1 79 theme conditions 104:113 arg1 practice 155:162 a routine industrial practice 134:162 a routine industrial practice used to improve recombinant protein production 134:209 The application of mild hypothermic conditions to cell culture is a routine industrial practice used to improve recombinant protein production. 25545631 0 80 theme monoclonal 33:42 arg1 glycosylation 53:65 monoclonal antibody glycosylation 33:65 monoclonal antibody glycosylation 33:65 How does mild hypothermia affect monoclonal antibody glycosylation? 25545631 3 81 theme mild 421:424 arg1 hypothermia 426:436 mild hypothermia 421:436 mild hypothermia 421:436 In this study, we investigated the impact of mild hypothermia on protein glycosylation. 25545631 10 82 theme mild 1485:1488 arg1 hypothermia 1490:1500 mild hypothermia 1485:1500 mild hypothermia 1485:1500 As a result of mild hypothermia, mAb glycosylation was shown to be affected by both NSD availability and glycosyltransferase expression. 25545631 7 83 theme stationary 1118:1127 arg1 phase 1129:1133 stationary phase 1118:1133 stationary phase of cultures with or without a temperature shift 1118:1181 To better understand CHO cell metabolism at 32°C, flux balance analysis (FBA) was carried out and constrained with exometabolite data from stationary phase of cultures with or without a temperature shift. 25545631 12 84 theme distribution 1839:1850 arg1 understanding 1803:1815 the understanding 1799:1815 the understanding of the metabolic flux distribution 1799:1850 Better feeding strategies can be developed based on the understanding of the metabolic flux distribution. 25545631 6 85 theme sugar 851:855 arg1 NSD 864:866 NSD 864:866 NSD 864:866 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 85 theme sugar 851:855 arg1 donor 857:861 nucleotide sugar donor 840:861 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 6 86 theme antibody 741:748 arg1 productivity 750:761 The specific antibody productivity 728:761 The specific antibody productivity 728:761 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 9 87 theme N-linked 1395:1402 arg1 glycan 1404:1409 N-linked glycan branching and elongation 1395:1434 glycan 1404:1409 Expression of the glycosyltransferases that are responsible for N-linked glycan branching and elongation were significantly lower at 32°C. 25545631 6 88 theme intracellular 826:838 arg1 concentrations 869:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 intracellular nucleotide sugar donor (NSD) concentrations 826:882 The specific antibody productivity increased by 25% at 32°C and was accompanied by a reduction in intracellular nucleotide sugar donor (NSD) concentrations and a decreased proportion of the more processed glycan structures on the mAb constant region. 25545631 4 89 theme late 605:608 arg1 phase 639:643 late exponential/early stationary phase 605:643 late exponential/early stationary phase 605:643 Chinese hamster ovary (CHO) cells expressing a monoclonal antibody (mAb) were cultured at 36.5°C and with a temperature shift to 32°C during late exponential/early stationary phase. 25545631 11 90 theme experimental/FBA 1620:1635 arg1 approach 1637:1644 The combined experimental/FBA approach 1607:1644 The combined experimental/FBA approach 1607:1644 The combined experimental/FBA approach generated insight as to how product glycosylation can be impacted by changes in culture temperature. 28264742 4 0 theme structures 667:676 arg1 complexity 640:649 The complexity 636:649 The complexity of carbohydrate structures 636:676 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 6 1 attach predominant 1488:1498 arg2 glycans 1326:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans 1154:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides 1154:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 1 attach predominant 1488:1498 arg1 MPS 1503:1505 MPS 1503:1505 MPS 1503:1505 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 2 theme MPS 1082:1084 arg1 microarrays 1095:1105 Both MPS and NMPS microarrays 1077:1105 microarrays 1095:1105 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 0 3 theme non-mating 80:89 arg1 periods 91:97 non-mating periods 80:97 non-mating periods 80:97 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 6 4 dep O-glycans 1176:1184 arg1 i 1173:1173 i 1173:1173 i 1173:1173 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 7 5 theme buffalo 1759:1765 arg1 sperm 1772:1776 buffalo bull sperm 1759:1776 buffalo bull sperm surface 1759:1784 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 1 6 theme non-mating 165:174 arg1 periods 176:182 non-mating periods 165:182 non-mating periods 165:182 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 7 dep dependent 1301:1309 arg1 dependent 1301:1309 dependent 1301:1309 dependent 1301:1309 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 7 dep dependent 1301:1309 arg1 ii 1313:1314 ii 1313:1314 ii 1313:1314 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 7 dep dependent 1301:1309 arg1 expression 1158:1167 the expression 1154:1167 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1154:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 8 theme l-Fucα1,2 1258:1266 arg1 Galβ1,3/4GlcNAcβ1 1268:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 9 theme GalNAcα1,3 1247:1256 arg1 Galβ1,3/4GlcNAcβ1 1268:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 4 10 theme cell 816:819 arg1 status 835:840 cell glycosylation status 816:840 cell glycosylation status 816:840 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 3 11 theme surface 519:525 arg1 pattern 541:547 the surface glycosylation pattern 515:547 the surface glycosylation pattern of mating period spermatozoa (MPS) 515:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 12 from predominant 1488:1498 arg1 MPS 1503:1505 MPS 1503:1505 MPS 1503:1505 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 1 13 theme autumn 258:263 arg1 beginning 245:253 beginning 245:253 beginning of autumn, respectively 245:277 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 14 theme O-linked 1317:1324 arg1 glycans 1326:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans 1154:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides 1154:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 2 15 theme Sperm 280:284 arg1 glycocalyx 286:295 Sperm glycocalyx 280:295 Sperm glycocalyx 280:295 Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. 28264742 4 16 dep makes 678:682 arg1 is 765:766 is 765:766 is now providing a new tool into the evaluation of cell glycosylation status 765:840 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 4 16 dep makes 678:682 arg1 challenging 699:709 challenging 699:709 challenging 699:709 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 3 17 theme glycosylation 527:539 arg1 pattern 541:547 the surface glycosylation pattern 515:547 the surface glycosylation pattern of mating period spermatozoa (MPS) 515:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 18 with O-glycans 1176:1184 arg1 Galβ1,3GalNAc 1229:1241 Galβ1,3GalNAc 1229:1241 Galβ1,3GalNAc 1229:1241 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 18 with O-glycans 1176:1184 arg1 Galβ1,3/4GlcNAcβ1 1268:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1247:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 18 with O-glycans 1176:1184 arg1 GalNAc 1221:1226 NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc 1191:1226 NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc 1191:1226 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 19 gly fucosylated 1455:1465 arg1 oligosaccharides 1467:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 4 20 theme status 835:840 arg1 evaluation 802:811 the evaluation 798:811 the evaluation of cell glycosylation status 798:840 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 3 21 from differences 500:510 arg1 pattern 541:547 the surface glycosylation pattern 515:547 the surface glycosylation pattern of mating period spermatozoa (MPS) 515:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 22 theme terminal 1607:1614 arg1 galactose 1616:1624 terminal galactose 1607:1624 terminal galactose 1607:1624 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 5 23 theme microarray 908:917 arg1 slides 919:924 microarray slides 908:924 microarray slides 908:924 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 3 24 theme non-mating 428:437 arg1 periods 439:445 non-mating periods 428:445 non-mating periods 428:445 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 25 theme Semen 397:401 arg1 quality 403:409 Semen quality 397:409 Semen quality 397:409 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 4 26 theme glycosylation 821:833 arg1 status 835:840 cell glycosylation status 816:840 cell glycosylation status 816:840 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 6 27 theme sialic 1590:1595 arg1 acids 1597:1601 α2,6 sialic acids 1585:1601 α2,6 sialic acids 1585:1601 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 5 28 theme biotinylated 956:967 arg1 lectins 969:975 12 biotinylated lectins 953:975 12 biotinylated lectins 953:975 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 6 29 theme α2,6 1585:1588 arg1 acids 1597:1601 α2,6 sialic acids 1585:1601 α2,6 sialic acids 1585:1601 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 3 30 theme mating 552:557 arg1 MPS 579:581 MPS 579:581 MPS 579:581 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 30 theme mating 552:557 arg1 spermatozoa 566:576 mating period spermatozoa 552:576 mating period spermatozoa (MPS) 552:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 5 31 theme novel 856:860 arg1 procedure 862:870 a novel procedure 854:870 a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner 854:1074 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 0 32 theme Differential 0:11 arg1 surface 13:19 Differential surface glycoprofile 0:32 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 0:98 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 7 33 theme innovative 1659:1668 arg1 method 1686:1691 this innovative cell microarray method 1654:1691 this innovative cell microarray method 1654:1691 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 7 34 theme specific 1714:1721 arg1 changes 1737:1743 specific glycosylation changes 1714:1743 specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods 1714:1825 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 3 35 theme period 559:564 arg1 MPS 579:581 MPS 579:581 MPS 579:581 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 35 theme period 559:564 arg1 spermatozoa 566:576 mating period spermatozoa 552:576 mating period spermatozoa (MPS) 552:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 1 36 theme late 199:202 arg1 autumn 204:209 late autumn 199:209 late autumn 199:209 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 37 from MPS 1503:1505 arg1 predominant 1488:1498 predominant 1488:1498 predominant 1488:1498 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 7 38 theme sperm 1772:1776 arg1 surface 1778:1784 buffalo bull sperm surface 1759:1784 buffalo bull sperm surface 1759:1784 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 6 39 link O-linked 1317:1324 arg1 glycans 1326:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans 1154:1332 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides 1154:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 4 40 theme microarray 745:754 arg1 approach 756:763 the high-throughput microarray approach 725:763 the high-throughput microarray approach 725:763 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 7 41 theme mating 1797:1802 arg1 periods 1819:1825 the mating and non-mating periods 1793:1825 periods 1819:1825 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 2 42 theme important 306:314 arg1 role 316:319 an important role 303:319 an important role 303:319 Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. 28264742 6 43 theme asialo 1359:1364 arg1 glycans 1375:1381 asialo N-linked glycans 1359:1381 asialo N-linked glycans terminating with Galβ1,4GlcNAc 1359:1412 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 7 44 theme glycosylation 1723:1735 arg1 changes 1737:1743 specific glycosylation changes 1714:1743 specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods 1714:1825 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 4 45 theme high-throughput 729:743 arg1 approach 756:763 the high-throughput microarray approach 725:763 the high-throughput microarray approach 725:763 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 3 46 theme seasonal 491:498 arg1 differences 500:510 seasonal differences 491:510 seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) 491:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 0 47 theme bull 45:48 arg1 spermatozoa 50:60 buffalo bull spermatozoa 37:60 buffalo bull spermatozoa 37:60 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 6 48 theme NeuNAcα2-3Galβ1,3 1191:1207 arg1 GalNAc 1221:1226 NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc 1191:1226 NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc 1191:1226 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 5 49 theme microarray 1057:1066 arg1 scanner 1068:1074 a microarray scanner 1055:1074 a microarray scanner 1055:1074 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 3 50 dep poorer 414:419 arg1 so 448:449 so 448:449 so 448:449 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 0 51 theme buffalo 37:43 arg1 spermatozoa 50:60 buffalo bull spermatozoa 37:60 buffalo bull spermatozoa 37:60 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 1 52 contain has 112:114 arg2 activity 140:147 a seasonal reproduction activity 116:147 a seasonal reproduction activity 116:147 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 1 52 contain has 112:114 arg1 buffalo 104:110 The buffalo 100:110 The buffalo 100:110 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 53 link α1,2-linked 1443:1453 arg1 oligosaccharides 1467:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 7 54 theme non-mating 1808:1817 arg1 periods 1819:1825 the mating and non-mating periods 1793:1825 periods 1819:1825 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 6 55 theme N-linked 1366:1373 arg1 glycans 1375:1381 asialo N-linked glycans 1359:1381 asialo N-linked glycans terminating with Galβ1,4GlcNAc 1359:1412 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 3 56 theme spermatozoa 566:576 arg1 pattern 541:547 the surface glycosylation pattern 515:547 the surface glycosylation pattern of mating period spermatozoa (MPS) 515:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 57 theme NMPS 1090:1093 arg1 microarrays 1095:1105 Both MPS and NMPS microarrays 1077:1105 microarrays 1095:1105 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 0 58 theme spermatozoa 50:60 arg1 surface 13:19 Differential surface glycoprofile 0:32 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 0:98 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 5 59 theme signal 1019:1024 arg1 intensity 1026:1034 then signal intensity 1014:1034 then signal intensity 1014:1034 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 6 60 theme complex 1544:1550 arg1 types 1552:1556 biantennary complex types 1532:1556 biantennary complex types 1532:1556 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 1 61 theme seasonal 118:125 arg1 activity 140:147 a seasonal reproduction activity 116:147 a seasonal reproduction activity 116:147 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 62 theme fucosylated 1455:1465 arg1 oligosaccharides 1467:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 5 63 theme streptavidin 996:1007 arg1 panel 944:948 a panel 942:948 a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin 942:1007 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 4 64 theme new 784:786 arg1 tool 788:791 a new tool 782:791 a new tool into the evaluation of cell glycosylation status 782:840 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 3 65 theme non-mating 598:607 arg1 NMPS 629:632 NMPS 629:632 NMPS 629:632 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 65 theme non-mating 598:607 arg1 spermatozoa 616:626 non-mating period spermatozoa 598:626 non-mating period spermatozoa (NMPS) 598:633 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 66 theme biantennary 1532:1542 arg1 types 1552:1556 biantennary complex types 1532:1556 biantennary complex types 1532:1556 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 1 67 theme reproduction 127:138 arg1 activity 140:147 a seasonal reproduction activity 116:147 a seasonal reproduction activity 116:147 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 6 68 theme α1,2-linked 1443:1453 arg1 oligosaccharides 1467:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 α1,2-linked fucosylated oligosaccharides 1443:1482 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 69 theme O-glycans 1176:1184 arg1 dependent 1301:1309 dependent 1301:1309 dependent 1301:1309 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 69 theme O-glycans 1176:1184 arg1 ii 1313:1314 ii 1313:1314 ii 1313:1314 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 69 theme O-glycans 1176:1184 arg1 expression 1158:1167 the expression 1154:1167 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1154:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 3 70 theme period 609:614 arg1 NMPS 629:632 NMPS 629:632 NMPS 629:632 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 70 theme period 609:614 arg1 spermatozoa 616:626 non-mating period spermatozoa 598:626 non-mating period spermatozoa (NMPS) 598:633 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 71 link N-linked 1366:1373 arg1 glycans 1375:1381 asialo N-linked glycans 1359:1381 asialo N-linked glycans terminating with Galβ1,4GlcNAc 1359:1412 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 4 72 theme carbohydrate 654:665 arg1 structures 667:676 carbohydrate structures 654:676 carbohydrate structures 654:676 The complexity of carbohydrate structures makes their analysis challenging, and recently the high-throughput microarray approach is now providing a new tool into the evaluation of cell glycosylation status. 28264742 1 73 theme late 230:233 arg1 spring 235:240 late spring 230:240 late spring 230:240 The buffalo has a seasonal reproduction activity with mating and non-mating periods occurring from late autumn to winter and from late spring to beginning of autumn, respectively. 28264742 7 74 theme bull 1767:1770 arg1 sperm 1772:1776 buffalo bull sperm 1759:1776 buffalo bull sperm surface 1759:1784 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 7 75 theme cell 1670:1673 arg1 method 1686:1691 this innovative cell microarray method 1654:1691 this innovative cell microarray method 1654:1691 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 28264742 5 76 theme Cy3-conjugated 981:994 arg1 streptavidin 996:1007 Cy3-conjugated streptavidin 981:1007 Cy3-conjugated streptavidin 981:1007 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 3 77 gly glycosylation 527:539 arg1 MPS 579:581 MPS 579:581 MPS 579:581 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 3 77 gly glycosylation 527:539 arg1 spermatozoa 566:576 mating period spermatozoa 552:576 mating period spermatozoa (MPS) 552:582 Semen quality is poorer during non-mating periods, so we aimed to evaluate if there were also seasonal differences in the surface glycosylation pattern of mating period spermatozoa (MPS) compared with non-mating period spermatozoa (NMPS). 28264742 6 78 dep mannose- 1519:1526 arg1 iii 1509:1511 iii 1509:1511 iii 1509:1511 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 78 dep mannose- 1519:1526 arg1 N-glycans 1558:1566 N-glycans 1558:1566 N-glycans terminating with α2,6 sialic acids and terminal galactose 1558:1624 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 5 79 theme lectins 969:975 arg1 panel 944:948 a panel 942:948 a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin 942:1007 We adopted a novel procedure in which spermatozoa was spotted on microarray slides, incubated with a panel of 12 biotinylated lectins and Cy3-conjugated streptavidin, and then signal intensity was detected using a microarray scanner. 28264742 6 80 dep glycans 1326:1332 arg1 dependent 1301:1309 dependent 1301:1309 dependent 1301:1309 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 80 dep glycans 1326:1332 arg1 ii 1313:1314 ii 1313:1314 ii 1313:1314 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 6 80 dep glycans 1326:1332 arg1 expression 1158:1167 the expression 1154:1167 the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 1154:1284 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 0 81 theme glycoprofile 21:32 arg1 surface 13:19 Differential surface glycoprofile 0:32 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 0:98 Differential surface glycoprofile of buffalo bull spermatozoa during mating and non-mating periods. 28264742 6 82 theme high 1514:1517 arg1 mannose- 1519:1526 (iii) high mannose- 1508:1526 (iii) high mannose- 1508:1526 Both MPS and NMPS microarrays reacted with all the lectins and revealed that the expression of (i) O-glycans with NeuNAcα2-3Galβ1,3(±NeuNAcα2-6)GalNAc, Galβ1,3GalNAc and GalNAcα1,3(l-Fucα1,2)Galβ1,3/4GlcNAcβ1 was not season dependent; (ii) O-linked glycans terminating with GalNAc, asialo N-linked glycans terminating with Galβ1,4GlcNAc, GlcNAc, as well as α1,6 and α1,2-linked fucosylated oligosaccharides was predominant in MPS; (iii) high mannose- and biantennary complex types N-glycans terminating with α2,6 sialic acids and terminal galactose were lower in MPS. 28264742 2 83 theme first 350:354 arg1 it 340:341 it 340:341 it 340:341 Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. 28264742 2 83 theme first 350:354 arg1 interface 356:364 the first interface 346:364 the first interface between sperm and environment 346:394 Sperm glycocalyx plays an important role in reproduction as it is the first interface between sperm and environment. 28264742 7 84 theme microarray 1675:1684 arg1 method 1686:1691 this innovative cell microarray method 1654:1691 this innovative cell microarray method 1654:1691 Overall, this innovative cell microarray method was able to identify specific glycosylation changes that occur on buffalo bull sperm surface during the mating and non-mating periods. 26114652 2 0 theme chromatographic 626:640 arg1 run 642:644 one chromatographic run 622:644 one chromatographic run 622:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 12 1 theme high 1999:2002 arg1 degree 2004:2009 a high degree 1997:2009 a high degree of consistency 1997:2024 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 12 2 dep types 2033:2037 arg1 the 2029:2031 the 2029:2031 the 2029:2031 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 7 3 theme Glycosylation 1223:1235 arg1 fingerprints 1237:1248 Glycosylation fingerprints 1223:1248 Glycosylation fingerprints 1223:1248 Glycosylation fingerprints were obtained using a fluorescently labelled antibody directed against human IgG Fc. 26114652 3 4 with HPAEC-PAD 736:744 arg1 standards 766:774 monosaccharide standards 751:774 monosaccharide standards 751:774 The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. 26114652 1 5 theme orthogonal 259:268 arg1 methods 286:292 three orthogonal chromatographic methods 253:292 three orthogonal chromatographic methods 253:292 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 13 6 theme separation 2312:2321 arg1 orthogonality 2289:2301 the orthogonality 2285:2301 the orthogonality of their separation and detection mechanisms 2285:2346 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 4 7 theme direct 903:908 arg1 quantitation 910:921 direct quantitation 903:921 direct quantitation 903:921 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 12 8 theme therapeutic 1968:1978 arg1 mAb 1980:1982 The therapeutic mAb 1964:1982 The therapeutic mAb 1964:1982 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 10 9 theme N-linked 1740:1747 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 9 theme N-linked 1740:1747 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 3 10 theme monosaccharide 751:764 arg1 standards 766:774 monosaccharide standards 751:774 monosaccharide standards 751:774 The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. 26114652 5 11 theme 2-AA- 928:932 arg1 versions 952:959 The 2-AA- and 2-AB-labelled versions 924:959 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel 924:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 1 12 theme therapeutic 199:209 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 12 theme therapeutic 199:209 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 10 13 theme asialo 1782:1787 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 13 theme asialo 1782:1787 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 4 14 with labelling 833:841 arg1 tags 874:877 two different fluorescent tags 848:877 two different fluorescent tags (2-AA and 2-AB) 848:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 14 with labelling 833:841 arg1 2-AB 889:892 2-AB 889:892 2-AB 889:892 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 14 with labelling 833:841 arg1 2-AA 880:883 2-AA 880:883 2-AA 880:883 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 9 15 theme complementary 1650:1662 arg1 information 1711:1721 complementary and corroboratory qualitative and quantitative information 1650:1721 information 1711:1721 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 1 16 theme antibody 222:229 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 16 theme antibody 222:229 arg1 lots 174:177 four lots 169:177 four lots of a marketed human therapeutic monoclonal antibody (mAb) 169:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 16 theme antibody 222:229 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 2 17 gly sialylated 600:609 arg1 glycans 611:617 typical neutral and sialylated glycans 580:617 typical neutral and sialylated glycans in one chromatographic run 580:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 2 18 theme sequential 557:566 arg1 analyses 568:575 sequential analyses 557:575 sequential analyses of typical neutral and sialylated glycans in one chromatographic run 557:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 8 19 theme proprietary 1407:1417 arg1 algorithm 1419:1427 a proprietary algorithm 1405:1427 a proprietary algorithm to obtain semi-quantitative "glycan structural class" information 1405:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 2 20 theme sialylated 600:609 arg1 glycans 611:617 typical neutral and sialylated glycans 580:617 typical neutral and sialylated glycans in one chromatographic run 580:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 10 21 gly core-fucosylated 1765:1780 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 21 gly core-fucosylated 1765:1780 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 13 22 theme results 2234:2240 arg1 agreement 2215:2223 the excellent quantitative agreement 2188:2223 the excellent quantitative agreement of their results 2188:2240 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 5 23 theme standard 980:987 arg1 panel 989:993 the same glycan standard panel 964:993 the same glycan standard panel 964:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 2 24 theme neutral 588:594 arg1 glycans 611:617 typical neutral and sialylated glycans 580:617 typical neutral and sialylated glycans in one chromatographic run 580:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 8 25 theme semi-quantitative 1439:1455 arg1 information 1483:1493 semi-quantitative "glycan structural class" information 1439:1493 semi-quantitative "glycan structural class" information 1439:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 3 26 theme monosaccharide 651:664 arg1 contents 666:673 The monosaccharide contents 647:673 The monosaccharide contents of these glycans following acid hydrolysis 647:716 The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. 26114652 1 27 theme N-linked 143:150 arg1 glycosylation 152:164 The N-linked glycosylation 139:164 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) 139:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 2 28 theme multi-stage 510:520 arg1 profile 545:551 a multi-stage linear gradient eluent profile 508:551 a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 508:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 4 29 dep tags 874:877 arg1 tags 874:877 two different fluorescent tags 848:877 two different fluorescent tags (2-AA and 2-AB) 848:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 29 dep tags 874:877 arg1 2-AB 889:892 2-AB 889:892 2-AB 889:892 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 29 dep tags 874:877 arg1 2-AA 880:883 2-AA 880:883 2-AA 880:883 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 8 30 theme glycan 1458:1463 arg1 information 1483:1493 semi-quantitative "glycan structural class" information 1439:1493 semi-quantitative "glycan structural class" information 1439:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 13 31 theme quantitative 2202:2213 arg1 agreement 2215:2223 the excellent quantitative agreement 2188:2223 the excellent quantitative agreement of their results 2188:2240 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 9 32 theme qualitative 1682:1692 arg1 information 1711:1721 complementary and corroboratory qualitative and quantitative information 1650:1721 information 1711:1721 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 8 33 theme class 1476:1480 arg1 information 1483:1493 semi-quantitative "glycan structural class" information 1439:1493 semi-quantitative "glycan structural class" information 1439:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 5 34 theme same 968:971 arg1 panel 989:993 the same glycan standard panel 964:993 the same glycan standard panel 964:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 2 35 theme profile 545:551 arg1 panel 473:477 a panel 471:477 a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 471:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 7 36 theme IgG 1327:1329 arg1 Fc 1331:1332 human IgG Fc 1321:1332 human IgG Fc 1321:1332 Glycosylation fingerprints were obtained using a fluorescently labelled antibody directed against human IgG Fc. 26114652 5 37 theme orthogonal 1048:1057 arg1 identification 1059:1072 orthogonal identification 1048:1072 orthogonal identification of mAb glycans 1048:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 2 38 theme gradient 529:536 arg1 profile 545:551 a multi-stage linear gradient eluent profile 508:551 a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 508:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 2 39 theme F. 418:419 arg1 glycans 428:434 PNGase F. Native glycans 411:434 PNGase F. Native glycans 411:434 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 0 40 theme monoclonal 118:127 arg1 antibody 129:136 a therapeutic monoclonal antibody 104:136 a therapeutic monoclonal antibody 104:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 6 41 theme intact 1171:1176 arg1 mAbs 1178:1181 the intact mAbs 1167:1181 the intact mAbs 1167:1181 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 9 42 theme Glycosylation 1496:1508 arg1 analyses 1510:1517 Glycosylation analyses 1496:1517 Glycosylation analyses 1496:1517 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 5 43 theme separation 1015:1024 arg1 profiles 1026:1033 distinctive separation profiles 1003:1033 distinctive separation profiles suitable for orthogonal identification of mAb glycans 1003:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 2 44 from glycans 611:617 arg1 run 642:644 one chromatographic run 622:644 one chromatographic run 622:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 0 45 theme orthogonal 14:23 arg1 methods 72:78 orthogonal chromatographic and lectin-affinity microarray methods 14:78 orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody 14:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 1 46 theme commercial 300:309 arg1 microarray 318:327 a commercial lectin microarray 298:327 a commercial lectin microarray 298:327 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 12 47 theme N-linked 2054:2061 arg1 glycans 2063:2069 N-linked glycans 2054:2069 N-linked glycans 2054:2069 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 5 48 theme suitable 1035:1042 arg1 profiles 1026:1033 distinctive separation profiles 1003:1033 distinctive separation profiles suitable for orthogonal identification of mAb glycans 1003:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 6 49 theme partial 1143:1149 arg1 denaturation 1151:1162 partial denaturation 1143:1162 partial denaturation of the intact mAbs 1143:1181 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 2 50 theme N-glycan 485:492 arg1 standards 494:502 21 N-glycan standards 482:502 21 N-glycan standards 482:502 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 3 51 theme glycans 684:690 arg1 contents 666:673 The monosaccharide contents 647:673 The monosaccharide contents of these glycans following acid hydrolysis 647:716 The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. 26114652 12 52 theme consistency 2014:2024 arg1 degree 2004:2009 a high degree 1997:2009 a high degree of consistency 1997:2024 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 2 53 from run 642:644 arg1 analyses 568:575 sequential analyses 557:575 sequential analyses of typical neutral and sialylated glycans in one chromatographic run 557:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 1 54 link N-linked 143:150 arg1 glycosylation 152:164 The N-linked glycosylation 139:164 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) 139:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 13 55 theme described 2150:2158 arg1 methods 2160:2166 The described methods 2146:2166 The described methods 2146:2166 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 0 56 theme microarray 61:70 arg1 methods 72:78 orthogonal chromatographic and lectin-affinity microarray methods 14:78 orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody 14:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 4 57 theme fluorescent 862:872 arg1 tags 874:877 two different fluorescent tags 848:877 two different fluorescent tags (2-AA and 2-AB) 848:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 57 theme fluorescent 862:872 arg1 2-AB 889:892 2-AB 889:892 2-AB 889:892 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 57 theme fluorescent 862:872 arg1 2-AA 880:883 2-AA 880:883 2-AA 880:883 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 9 58 theme mAb 1531:1533 arg1 lots 1535:1538 the four mAb lots 1522:1538 the four mAb lots 1522:1538 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 5 59 theme glycans 1081:1087 arg1 identification 1059:1072 orthogonal identification 1048:1072 orthogonal identification of mAb glycans 1048:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 6 60 theme Fc 1209:1210 arg1 N-glycans 1212:1220 the sequestered Fc N-glycans 1193:1220 the sequestered Fc N-glycans 1193:1220 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 10 61 theme predominant 1728:1738 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 61 theme predominant 1728:1738 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 3 62 theme acid 702:705 arg1 hydrolysis 707:716 acid hydrolysis 702:716 acid hydrolysis 702:716 The monosaccharide contents of these glycans following acid hydrolysis were confirmed by HPAEC-PAD with monosaccharide standards. 26114652 10 63 link N-linked 1740:1747 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 63 link N-linked 1740:1747 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 1 64 theme marketed 184:191 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 64 theme marketed 184:191 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 13 65 theme mechanisms 2337:2346 arg1 orthogonality 2289:2301 the orthogonality 2285:2301 the orthogonality of their separation and detection mechanisms 2285:2346 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 2 66 from analyses 568:575 arg1 run 642:644 one chromatographic run 622:644 one chromatographic run 622:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 10 67 theme core-fucosylated 1765:1780 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 67 theme core-fucosylated 1765:1780 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 1 68 theme chromatographic 270:284 arg1 methods 286:292 three orthogonal chromatographic methods 253:292 three orthogonal chromatographic methods 253:292 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 2 69 theme glycans 611:617 arg1 analyses 568:575 sequential analyses 557:575 sequential analyses of typical neutral and sialylated glycans in one chromatographic run 557:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 11 70 theme bisected 1888:1895 arg1 glycans 1897:1903 afucosyl and bisected glycans 1875:1903 afucosyl and bisected glycans 1875:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 11 71 theme afucosyl 1875:1882 arg1 glycans 1897:1903 afucosyl and bisected glycans 1875:1903 afucosyl and bisected glycans 1875:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 1 72 theme human 193:197 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 72 theme human 193:197 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 12 73 theme analysis 2119:2126 arg1 methods 2128:2134 all four analysis methods 2110:2134 all four analysis methods (<6% CV) 2110:2143 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 12 73 theme analysis 2119:2126 arg1 % 2139:2139 <6% CV 2137:2142 <6% CV 2137:2142 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 10 74 theme diantennary 1789:1799 arg1 structures 1801:1810 core-fucosylated asialo diantennary structures 1765:1810 core-fucosylated asialo diantennary structures 1765:1810 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 10 74 theme diantennary 1789:1799 arg1 structures 1749:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures 1724:1758 The predominant N-linked structures were core-fucosylated asialo diantennary structures with varying galactosylation. 26114652 1 75 theme monoclonal 211:220 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 75 theme monoclonal 211:220 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 9 76 theme quantitative 1698:1709 arg1 information 1711:1721 complementary and corroboratory qualitative and quantitative information 1650:1721 information 1711:1721 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 1 77 gly glycosylation 152:164 arg1 antibody 222:229 therapeutic monoclonal antibody 199:229 a marketed human therapeutic monoclonal antibody (mAb) 182:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 77 gly glycosylation 152:164 arg1 lots 174:177 four lots 169:177 four lots of a marketed human therapeutic monoclonal antibody (mAb) 169:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 1 77 gly glycosylation 152:164 arg1 mAb 232:234 mAb 232:234 mAb 232:234 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 5 78 theme 2-AB-labelled 938:950 arg1 versions 952:959 The 2-AA- and 2-AB-labelled versions 924:959 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel 924:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 9 79 theme different 1619:1627 arg1 principles 1629:1638 different principles 1619:1638 different principles 1619:1638 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 6 80 with profiling 1097:1105 arg1 microarray 1123:1132 the lectin microarray 1112:1132 the lectin microarray 1112:1132 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 7 81 theme human 1321:1325 arg1 Fc 1331:1332 human IgG Fc 1321:1332 human IgG Fc 1321:1332 Glycosylation fingerprints were obtained using a fluorescently labelled antibody directed against human IgG Fc. 26114652 8 82 from fingerprints 1369:1380 arg1 intensities 1348:1358 Fluorescence intensities 1335:1358 Fluorescence intensities from the fingerprints 1335:1380 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 5 83 theme glycan 973:978 arg1 panel 989:993 the same glycan standard panel 964:993 the same glycan standard panel 964:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 8 84 theme Fluorescence 1335:1346 arg1 intensities 1348:1358 Fluorescence intensities 1335:1358 Fluorescence intensities from the fingerprints 1335:1380 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 5 85 theme panel 989:993 arg1 versions 952:959 The 2-AA- and 2-AB-labelled versions 924:959 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel 924:993 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 6 86 theme lectin 1116:1121 arg1 microarray 1123:1132 the lectin microarray 1112:1132 the lectin microarray 1112:1132 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 2 87 theme typical 580:586 arg1 glycans 611:617 typical neutral and sialylated glycans 580:617 typical neutral and sialylated glycans in one chromatographic run 580:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 0 88 theme glycan 84:89 arg1 profiling 91:99 glycan profiling 84:99 glycan profiling of a therapeutic monoclonal antibody 84:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 9 89 theme corroboratory 1668:1680 arg1 information 1711:1721 complementary and corroboratory qualitative and quantitative information 1650:1721 information 1711:1721 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 1 90 theme lots 174:177 arg1 glycosylation 152:164 The N-linked glycosylation 139:164 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) 139:235 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 6 91 theme Glycan 1090:1095 arg1 profiling 1097:1105 Glycan profiling 1090:1105 Glycan profiling with the lectin microarray 1090:1132 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 2 92 theme standards 494:502 arg1 panel 473:477 a panel 471:477 a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 471:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 8 93 theme structural 1465:1474 arg1 information 1483:1493 semi-quantitative "glycan structural class" information 1439:1493 semi-quantitative "glycan structural class" information 1439:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 0 94 theme therapeutic 106:116 arg1 antibody 129:136 a therapeutic monoclonal antibody 104:136 a therapeutic monoclonal antibody 104:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 2 95 theme eluent 538:543 arg1 profile 545:551 a multi-stage linear gradient eluent profile 508:551 a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 508:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 8 96 theme " 1481:1481 arg1 information 1483:1493 semi-quantitative "glycan structural class" information 1439:1493 semi-quantitative "glycan structural class" information 1439:1493 Fluorescence intensities from the fingerprints were deconvoluted with a proprietary algorithm to obtain semi-quantitative "glycan structural class" information. 26114652 4 97 theme stoichiometric 818:831 arg1 labelling 833:841 stoichiometric labelling 818:841 stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) 818:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 0 98 theme antibody 129:136 arg1 profiling 91:99 glycan profiling 84:99 glycan profiling of a therapeutic monoclonal antibody 84:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 6 99 theme mAbs 1178:1181 arg1 denaturation 1151:1162 partial denaturation 1143:1162 partial denaturation of the intact mAbs 1143:1181 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 2 100 theme linear 522:527 arg1 profile 545:551 a multi-stage linear gradient eluent profile 508:551 a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run 508:644 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 13 101 theme detection 2327:2335 arg1 mechanisms 2337:2346 their separation and detection mechanisms 2306:2346 mechanisms 2337:2346 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 2 102 theme PNGase 411:416 arg1 glycans 428:434 PNGase F. Native glycans 411:434 PNGase F. Native glycans 411:434 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 7 103 theme labelled 1286:1293 arg1 antibody 1295:1302 a fluorescently labelled antibody 1270:1302 a fluorescently labelled antibody directed against human IgG Fc 1270:1332 Glycosylation fingerprints were obtained using a fluorescently labelled antibody directed against human IgG Fc. 26114652 13 104 theme excellent 2192:2200 arg1 agreement 2215:2223 the excellent quantitative agreement 2188:2223 the excellent quantitative agreement of their results 2188:2240 The described methods are co-supported by the excellent quantitative agreement of their results, which is particularly notable considering the orthogonality of their separation and detection mechanisms. 26114652 5 105 theme distinctive 1003:1013 arg1 profiles 1026:1033 distinctive separation profiles 1003:1033 distinctive separation profiles suitable for orthogonal identification of mAb glycans 1003:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 0 106 theme chromatographic 25:39 arg1 methods 72:78 orthogonal chromatographic and lectin-affinity microarray methods 14:78 orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody 14:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 12 107 theme glycans 2063:2069 arg1 amounts 2043:2049 amounts 2043:2049 amounts 2043:2049 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 12 107 theme glycans 2063:2069 arg1 glycans 2063:2069 N-linked glycans 2054:2069 N-linked glycans 2054:2069 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 12 107 theme glycans 2063:2069 arg1 types 2033:2037 types 2033:2037 types 2033:2037 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 0 108 theme lectin-affinity 45:59 arg1 methods 72:78 orthogonal chromatographic and lectin-affinity microarray methods 14:78 orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody 14:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 1 109 theme lectin 311:316 arg1 microarray 318:327 a commercial lectin microarray 298:327 a commercial lectin microarray 298:327 The N-linked glycosylation of four lots of a marketed human therapeutic monoclonal antibody (mAb) was assessed by three orthogonal chromatographic methods and a commercial lectin microarray. 26114652 12 110 link N-linked 2054:2061 arg1 glycans 2063:2069 N-linked glycans 2054:2069 N-linked glycans 2054:2069 The therapeutic mAb demonstrated a high degree of consistency in the types and amounts of N-linked glycans in the four lots (<6% CV), and between all four analysis methods (<6% CV). 26114652 11 111 theme glycans 1897:1903 arg1 glycans 1897:1903 afucosyl and bisected glycans 1875:1903 afucosyl and bisected glycans 1875:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 11 111 theme glycans 1897:1903 arg1 amounts 1864:1870 trace amounts 1858:1870 trace amounts of afucosyl and bisected glycans 1858:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 2 112 theme Native 421:426 arg1 glycans 428:434 PNGase F. Native glycans 411:434 PNGase F. Native glycans 411:434 For chromatography, the N-glycans were removed enzymatically from the mAbs using PNGase F. Native glycans were determined by HPAEC-PAD using a panel of 21 N-glycan standards and a multi-stage linear gradient eluent profile for sequential analyses of typical neutral and sialylated glycans in one chromatographic run. 26114652 0 113 theme methods 72:78 arg1 Comparison 0:9 Comparison 0:9 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody 0:136 Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody. 26114652 4 114 theme Glycosylation 777:789 arg1 analysis 791:798 Glycosylation analysis 777:798 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) 777:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 5 115 theme mAb 1077:1079 arg1 glycans 1081:1087 mAb glycans 1077:1087 mAb glycans 1077:1087 The 2-AA- and 2-AB-labelled versions of the same glycan standard panel yielded distinctive separation profiles suitable for orthogonal identification of mAb glycans. 26114652 11 116 theme detectable 1913:1922 arg1 sialylation 1924:1934 no detectable sialylation 1910:1934 no detectable sialylation by any of the four methods 1910:1961 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 4 117 theme different 852:860 arg1 tags 874:877 two different fluorescent tags 848:877 two different fluorescent tags (2-AA and 2-AB) 848:893 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 117 theme different 852:860 arg1 2-AB 889:892 2-AB 889:892 2-AB 889:892 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 4 117 theme different 852:860 arg1 2-AA 880:883 2-AA 880:883 2-AA 880:883 Glycosylation analysis by HILIC-FD after stoichiometric labelling with two different fluorescent tags (2-AA and 2-AB) enabled direct quantitation. 26114652 9 118 theme lots 1535:1538 arg1 analyses 1510:1517 Glycosylation analyses 1496:1517 Glycosylation analyses 1496:1517 Glycosylation analyses of the four mAb lots by these four methods, which separate and detect oligosaccharides according to different principles, provided complementary and corroboratory qualitative and quantitative information. 26114652 6 119 theme sequestered 1197:1207 arg1 N-glycans 1212:1220 the sequestered Fc N-glycans 1193:1220 the sequestered Fc N-glycans 1193:1220 Glycan profiling with the lectin microarray required partial denaturation of the intact mAbs to expose the sequestered Fc N-glycans. 26114652 11 120 theme trace 1858:1862 arg1 glycans 1897:1903 afucosyl and bisected glycans 1875:1903 afucosyl and bisected glycans 1875:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26114652 11 120 theme trace 1858:1862 arg1 amounts 1864:1870 trace amounts 1858:1870 trace amounts of afucosyl and bisected glycans 1858:1903 There were also trace amounts of afucosyl and bisected glycans, but no detectable sialylation by any of the four methods. 26170448 3 0 gly glycoprotein 433:444 arg1 glycoprotein 433:444 the S-layer glycoprotein 421:444 the S-layer glycoprotein 421:444 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 1 1 theme members 160:166 arg1 members 160:166 members 160:166 members of the Halobacteriaceae, which displays particularly low restriction activity 160:244 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 1 1 theme members 160:166 arg1 one 153:155 one 153:155 one 153:155 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 9 2 theme nuclear 951:957 arg1 resonance 968:976 nuclear magnetic resonance 951:976 nuclear magnetic resonance 951:976 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 6 3 gly glycoproteins 635:647 arg1 glycoproteins 635:647 two different glycoproteins 621:647 two different glycoproteins which shared highly similar amino acid sequences 621:696 The S-layer was found containing two different glycoproteins which shared highly similar amino acid sequences. 26170448 11 4 theme new 1547:1549 arg1 model 1551:1555 a new model 1545:1555 a new model 1545:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 11 4 theme new 1547:1549 arg1 hispanica 1532:1540 H. hispanica 1529:1540 H. hispanica 1529:1540 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 8 5 theme S-layer 864:870 arg1 glycoproteins 872:884 these two S-layer glycoproteins 854:884 these two S-layer glycoproteins 854:884 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 0 6 theme archaeon 98:105 arg1 hispanica 118:126 the halophilic archaeon Haloarcula hispanica 83:126 the halophilic archaeon Haloarcula hispanica 83:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 0 7 from Identification 0:13 arg1 hispanica 118:126 the halophilic archaeon Haloarcula hispanica 83:126 the halophilic archaeon Haloarcula hispanica 83:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 11 8 theme diverse 1469:1475 arg1 pathways 1499:1506 the diverse protein glycosylation pathways 1465:1506 the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model 1465:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 0 9 theme halophilic 87:96 arg1 hispanica 118:126 the halophilic archaeon Haloarcula hispanica 83:126 the halophilic archaeon Haloarcula hispanica 83:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 8 10 link O-linked 818:825 arg1 glycans 827:833 O-linked glycans 818:833 O-linked glycans 818:833 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 10 11 theme glycosidase 1362:1372 arg1 treatment 1374:1382 glycosidase treatment 1362:1382 glycosidase treatment 1362:1382 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 3 12 gly glycosylated 407:418 arg1 glycoprotein 433:444 the S-layer glycoprotein 421:444 the S-layer glycoprotein 421:444 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 7 13 gly glycoproteins 738:750 arg1 glycoproteins 738:750 these two S-layer glycoproteins 720:750 these two S-layer glycoproteins 720:750 The genes coding for these two S-layer glycoproteins were found next to each other in the genome. 26170448 4 14 dep proteins 527:534 arg1 study 508:512 this study 503:512 this study 503:512 In this study, the S-layer proteins of Har. 26170448 9 15 theme Da 1052:1053 arg1 residue 1055:1061 a 225 Da residue 1046:1061 a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified 1046:1213 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 0 16 theme Haloarcula 107:116 arg1 hispanica 118:126 the halophilic archaeon Haloarcula hispanica 83:126 the halophilic archaeon Haloarcula hispanica 83:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 9 17 theme magnetic 959:966 arg1 resonance 968:976 nuclear magnetic resonance 951:976 nuclear magnetic resonance 951:976 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 10 18 theme monosaccharide 1322:1335 arg1 analysis 1349:1356 monosaccharide composition analysis 1322:1356 monosaccharide composition analysis 1322:1356 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 1 19 theme Halobacteriaceae 175:190 arg1 members 160:166 members 160:166 members of the Halobacteriaceae, which displays particularly low restriction activity 160:244 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 6 20 theme different 625:633 arg1 glycoproteins 635:647 two different glycoproteins 621:647 two different glycoproteins which shared highly similar amino acid sequences 621:696 The S-layer was found containing two different glycoproteins which shared highly similar amino acid sequences. 26170448 8 21 theme structural 890:899 arg1 determination 901:913 structural determination 890:913 structural determination 890:913 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 11 22 theme O-glycan 1417:1424 arg1 helpful 1444:1450 helpful 1444:1450 helpful 1444:1450 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 11 22 theme O-glycan 1417:1424 arg1 determination 1389:1401 The determination 1385:1401 The determination of the N- and O-glycan structure 1385:1434 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 3 23 theme glycosylated 407:418 arg1 glycoprotein 433:444 the S-layer glycoprotein 421:444 the S-layer glycoprotein 421:444 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 11 24 dep N- 1410:1411 arg1 the 1406:1408 the 1406:1408 the 1406:1408 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 11 24 dep N- 1410:1411 arg1 structure 1426:1434 structure 1426:1434 structure 1426:1434 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 0 25 theme S-layer 22:28 arg1 glycoproteins 30:42 the S-layer glycoproteins 18:42 the S-layer glycoproteins 18:42 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 10 26 theme composition 1337:1347 arg1 analysis 1349:1356 monosaccharide composition analysis 1322:1356 monosaccharide composition analysis 1322:1356 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 3 27 theme S-layer 373:379 arg1 protein 381:387 hispanica S-layer protein 363:387 hispanica S-layer protein 363:387 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 10 28 theme Glcα-1,4-Gal 1261:1272 arg1 O-glycan 1229:1236 the O-glycan 1225:1236 the O-glycan 1225:1236 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 10 28 theme Glcα-1,4-Gal 1261:1272 arg1 disaccharide 1274:1285 a Glcα-1,4-Gal disaccharide 1259:1285 a Glcα-1,4-Gal disaccharide 1259:1285 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 8 29 theme O-linked 818:825 arg1 glycans 827:833 O-linked glycans 818:833 O-linked glycans 818:833 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 1 30 theme tractable 292:300 arg1 haloarchaea 302:312 the most tractable haloarchaea 283:312 the most tractable haloarchaea for archaeal genetic research 283:342 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 9 31 theme first 1135:1139 arg1 2-amino-6-sulfo-2 1081:1097 a 2-amino-6-sulfo-2 1079:1097 a 2-amino-6-sulfo-2 1079:1097 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 9 31 theme first 1135:1139 arg1 time 1141:1144 the first time that a naturally occurring form of sulfoquinovosamine was identified 1131:1213 the first time that a naturally occurring form of sulfoquinovosamine was identified 1131:1213 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 3 32 theme S-layer 425:431 arg1 glycoprotein 433:444 the S-layer glycoprotein 421:444 the S-layer glycoprotein 421:444 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 1 33 theme haloarchaea 302:312 arg1 haloarchaea 302:312 the most tractable haloarchaea 283:312 the most tractable haloarchaea for archaeal genetic research 283:342 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 1 33 theme haloarchaea 302:312 arg1 one 276:278 one 276:278 one 276:278 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 7 34 theme S-layer 730:736 arg1 glycoproteins 738:750 these two S-layer glycoproteins 720:750 these two S-layer glycoproteins 720:750 The genes coding for these two S-layer glycoproteins were found next to each other in the genome. 26170448 11 35 theme H. 1529:1530 arg1 model 1551:1555 a new model 1545:1555 a new model 1545:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 11 35 theme H. 1529:1530 arg1 hispanica 1532:1540 H. hispanica 1529:1540 H. hispanica 1529:1540 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 1 36 theme low 221:223 arg1 activity 237:244 particularly low restriction activity 208:244 particularly low restriction activity 208:244 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 0 37 theme glycoproteins 30:42 arg1 Identification 0:13 Identification 0:13 Identification of the S-layer glycoproteins 0:42 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 0 37 theme glycoproteins 30:42 arg1 glycans 72:78 their covalently linked glycans 48:78 their covalently linked glycans in the halophilic archaeon Haloarcula hispanica 48:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 4 38 theme Har 539:541 arg1 proteins 527:534 the S-layer proteins 515:534 the S-layer proteins of Har 515:541 In this study, the S-layer proteins of Har. 26170448 9 39 theme branched 1012:1019 arg1 N-glycan 983:990 the N-glycan 979:990 the N-glycan 979:990 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 9 39 theme branched 1012:1019 arg1 trisaccharide 1021:1033 a branched trisaccharide 1010:1033 a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified 1010:1213 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 0 40 from glycans 72:78 arg1 hispanica 118:126 the halophilic archaeon Haloarcula hispanica 83:126 the halophilic archaeon Haloarcula hispanica 83:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 7 41 located found 757:761 arg1 genome 789:794 the genome 785:794 the genome 785:794 The genes coding for these two S-layer glycoproteins were found next to each other in the genome. 26170448 7 41 located found 757:761 arg2 genes 703:707 The genes 699:707 The genes coding for these two S-layer glycoproteins 699:750 The genes coding for these two S-layer glycoproteins were found next to each other in the genome. 26170448 1 42 theme restriction 225:235 arg1 activity 237:244 particularly low restriction activity 208:244 particularly low restriction activity 208:244 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 11 43 theme N- 1410:1411 arg1 helpful 1444:1450 helpful 1444:1450 helpful 1444:1450 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 11 43 theme N- 1410:1411 arg1 determination 1389:1401 The determination 1385:1401 The determination of the N- and O-glycan structure 1385:1434 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 1 44 theme archaeal 318:325 arg1 research 335:342 archaeal genetic research 318:342 archaeal genetic research 318:342 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 8 45 attach released 840:847 arg1 glycoproteins 872:884 these two S-layer glycoproteins 854:884 these two S-layer glycoproteins 854:884 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 8 45 attach released 840:847 arg2 N- 811:812 the N- 807:812 the N- 807:812 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 8 45 attach released 840:847 arg2 glycans 827:833 O-linked glycans 818:833 O-linked glycans 818:833 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 4 46 theme S-layer 519:525 arg1 proteins 527:534 the S-layer proteins 515:534 the S-layer proteins of Har 515:541 In this study, the S-layer proteins of Har. 26170448 1 47 theme genetic 327:333 arg1 research 335:342 archaeal genetic research 318:342 archaeal genetic research 318:342 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 3 48 theme hispanica 363:371 arg1 protein 381:387 hispanica S-layer protein 363:387 hispanica S-layer protein 363:387 hispanica S-layer protein has been reported glycosylated, the S-layer glycoprotein and its glycosylation have not been investigated yet. 26170448 1 49 theme Haloarcula 129:138 arg1 hispanica 140:148 Haloarcula hispanica 129:148 Haloarcula hispanica 129:148 Haloarcula hispanica is one of members of the Halobacteriaceae, which displays particularly low restriction activity and is therefore important as one of the most tractable haloarchaea for archaeal genetic research. 26170448 9 50 contain containing 1035:1044 arg2 residue 1055:1061 a 225 Da residue 1046:1061 a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified 1046:1213 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 9 50 contain containing 1035:1044 arg1 N-glycan 983:990 the N-glycan 979:990 the N-glycan 979:990 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 9 50 contain containing 1035:1044 arg1 trisaccharide 1021:1033 a branched trisaccharide 1010:1033 a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified 1010:1213 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 0 51 gly glycoproteins 30:42 arg1 glycoproteins 30:42 the S-layer glycoproteins 18:42 the S-layer glycoproteins 18:42 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 0 52 link linked 65:70 arg1 glycans 72:78 their covalently linked glycans 48:78 their covalently linked glycans in the halophilic archaeon Haloarcula hispanica 48:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 6 53 theme acid 683:686 arg1 sequences 688:696 highly similar amino acid sequences 662:696 highly similar amino acid sequences 662:696 The S-layer was found containing two different glycoproteins which shared highly similar amino acid sequences. 26170448 10 54 theme mass 1290:1293 arg1 spectrometry 1295:1306 mass spectrometry 1290:1306 mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment 1290:1382 Besides, the O-glycan was characterized as a Glcα-1,4-Gal disaccharide by mass spectrometry combined with monosaccharide composition analysis and glycosidase treatment. 26170448 9 55 theme occurring 1163:1171 arg1 form 1173:1176 a naturally occurring form 1151:1176 a naturally occurring form of sulfoquinovosamine 1151:1198 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 8 56 gly glycoproteins 872:884 arg1 glycoproteins 872:884 these two S-layer glycoproteins 854:884 these two S-layer glycoproteins 854:884 Moreover, the N- and O-linked glycans were released from these two S-layer glycoproteins for structural determination. 26170448 11 57 from pathways 1499:1506 arg1 archaea 1511:1517 archaea 1511:1517 archaea utilizing H. hispanica as a new model 1511:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 6 58 theme amino 677:681 arg1 sequences 688:696 highly similar amino acid sequences 662:696 highly similar amino acid sequences 662:696 The S-layer was found containing two different glycoproteins which shared highly similar amino acid sequences. 26170448 11 59 theme glycosylation 1485:1497 arg1 pathways 1499:1506 the diverse protein glycosylation pathways 1465:1506 the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model 1465:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 26170448 9 60 theme mass 929:932 arg1 spectrometry 934:945 the mass spectrometry 925:945 the mass spectrometry 925:945 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 6 61 theme similar 669:675 arg1 sequences 688:696 highly similar amino acid sequences 662:696 highly similar amino acid sequences 662:696 The S-layer was found containing two different glycoproteins which shared highly similar amino acid sequences. 26170448 0 62 theme linked 65:70 arg1 glycans 72:78 their covalently linked glycans 48:78 their covalently linked glycans in the halophilic archaeon Haloarcula hispanica 48:126 Identification of the S-layer glycoproteins and their covalently linked glycans in the halophilic archaeon Haloarcula hispanica. 26170448 9 63 theme sulfoquinovosamine 1181:1198 arg1 form 1173:1176 a naturally occurring form 1151:1176 a naturally occurring form of sulfoquinovosamine 1151:1198 Based on the mass spectrometry and nuclear magnetic resonance, the N-glycan was determined as a branched trisaccharide containing a 225 Da residue corresponded to a 2-amino-6-sulfo-2, 6-dideoxy-quinovose, which was the first time that a naturally occurring form of sulfoquinovosamine was identified. 26170448 11 64 theme protein 1477:1483 arg1 pathways 1499:1506 the diverse protein glycosylation pathways 1465:1506 the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model 1465:1555 The determination of the N- and O-glycan structure will be helpful for studying the diverse protein glycosylation pathways in archaea utilizing H. hispanica as a new model. 27951443 2 0 theme containing 573:582 arg1 products 584:591 all 2-acetamido-2-deoxy-D-galactopyranoside containing products 529:591 all 2-acetamido-2-deoxy-D-galactopyranoside containing products 529:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 1 1 theme fucosylated 362:372 arg1 FCS 396:398 FCS 396:398 FCS 396:398 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 1 theme fucosylated 362:372 arg1 sulfates 386:393 fucosylated chondroitin sulfates 362:393 fucosylated chondroitin sulfates (FCS) from sea cucumbers 362:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 3 2 theme bromide 894:900 arg1 donor 902:906 structurally related fucosyl bromide donor 865:906 structurally related fucosyl bromide donor 865:906 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 1 3 theme Unsubstituted 132:144 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 3 theme Unsubstituted 132:144 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 3 theme Unsubstituted 132:144 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 5 4 theme O-sulfation 1289:1299 arg1 effects 1278:1284 the spectral effects 1265:1284 the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS 1265:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 0 5 theme chondroitin 69:79 arg1 sulfates 81:88 fucosylated chondroitin sulfates 57:88 fucosylated chondroitin sulfates 57:88 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 1 6 theme chondroitin 374:384 arg1 FCS 396:398 FCS 396:398 FCS 396:398 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 6 theme chondroitin 374:384 arg1 sulfates 386:393 fucosylated chondroitin sulfates 362:393 fucosylated chondroitin sulfates (FCS) from sea cucumbers 362:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 7 gly 6-O-α-L-fucosylated 150:168 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 7 gly 6-O-α-L-fucosylated 150:168 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 7 gly 6-O-α-L-fucosylated 150:168 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 8 theme sulfates 386:393 arg1 fragments 349:357 the fragments 345:357 the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 345:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 5 9 theme target 1162:1167 arg1 products 1169:1176 the target products 1158:1176 the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS 1158:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 3 10 theme 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl 687:727 arg1 trichloracetimidate 729:747 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate 687:747 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf 687:773 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 11 theme donor 902:906 arg1 use 858:860 the use 854:860 the use of structurally related fucosyl bromide donor with promotion by Bu4NBr 854:931 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 12 gly 6-O-glycosylation 606:622 arg1 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 634:682 propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 627:682 propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 627:682 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 0 13 gly fucosylated 57:67 arg1 sulfates 81:88 fucosylated chondroitin sulfates 57:88 fucosylated chondroitin sulfates 57:88 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 1 14 gly fucosylated 362:372 arg1 FCS 396:398 FCS 396:398 FCS 396:398 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 14 gly fucosylated 362:372 arg1 sulfates 386:393 fucosylated chondroitin sulfates 362:393 fucosylated chondroitin sulfates (FCS) from sea cucumbers 362:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 0 15 theme Fuc 102:104 arg1 derivatives 119:129 Fuc(1 → 6)GalNAc derivatives 102:129 Fuc(1 → 6)GalNAc derivatives 102:129 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 4 16 theme O-protections 1034:1046 arg1 removal 1012:1018 Selective removal 1002:1018 Selective removal of orthogonal O-protections 1002:1046 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 3 17 theme propyl 627:632 arg1 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 634:682 propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 627:682 propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 627:682 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 1 18 theme 6-O-α-L-fucosylated 150:168 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 18 theme 6-O-α-L-fucosylated 150:168 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 18 theme 6-O-α-L-fucosylated 150:168 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 19 from cucumbers 410:418 arg1 FCS 396:398 FCS 396:398 FCS 396:398 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 19 from cucumbers 410:418 arg1 sulfates 386:393 fucosylated chondroitin sulfates 362:393 fucosylated chondroitin sulfates (FCS) from sea cucumbers 362:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 19 from cucumbers 410:418 arg1 fragments 349:357 the fragments 345:357 the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 345:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 3 20 theme fucosyl 886:892 arg1 donor 902:906 structurally related fucosyl bromide donor 865:906 structurally related fucosyl bromide donor 865:906 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 4 21 theme orthogonal 1023:1032 arg1 O-protections 1034:1046 orthogonal O-protections 1023:1046 orthogonal O-protections 1023:1046 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 3 22 theme 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside 634:682 arg1 6-O-glycosylation 606:622 6-O-glycosylation 606:622 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf 606:773 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 1 23 theme propyl 170:175 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 23 theme propyl 170:175 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 23 theme propyl 170:175 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 2 24 theme 2-acetamido-2-deoxy-D-galactopyranoside 533:571 arg1 products 584:591 all 2-acetamido-2-deoxy-D-galactopyranoside containing products 529:591 all 2-acetamido-2-deoxy-D-galactopyranoside containing products 529:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 4 25 theme subsequent 1058:1067 arg1 O-sulfation 1069:1079 subsequent O-sulfation 1058:1079 subsequent O-sulfation both at the GalNAc and Fuc units 1058:1112 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 4 26 dep units 1108:1112 arg1 both 1081:1084 both 1081:1084 both 1081:1084 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 5 27 theme blocking 1134:1141 arg1 groups 1143:1148 blocking groups 1134:1148 blocking groups 1134:1148 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 5 28 theme groups 1143:1148 arg1 removal 1123:1129 Further removal 1115:1129 Further removal of blocking groups 1115:1148 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 1 29 from fragments 349:357 arg1 cucumbers 410:418 sea cucumbers 406:418 sea cucumbers 406:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 2 30 theme key 492:494 arg1 precursor 496:504 a key precursor 490:504 a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products 490:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 2 30 theme key 492:494 arg1 2-deoxy-2-N-phthalimido-D-glucopyranose 438:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 5 31 theme natural 1393:1399 arg1 FCS 1401:1403 natural FCS 1393:1403 natural FCS 1393:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 0 32 theme 1 → 6 106:110 arg1 derivatives 119:129 Fuc(1 → 6)GalNAc derivatives 102:129 Fuc(1 → 6)GalNAc derivatives 102:129 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 0 33 theme NMR 14:16 arg1 analysis 18:25 NMR analysis 14:25 NMR analysis 14:25 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 5 34 theme FCS 1401:1403 arg1 analysis 1381:1388 computer assisted structural analysis 1352:1388 computer assisted structural analysis of natural FCS 1352:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 1 35 theme sea 406:408 arg1 cucumbers 410:418 sea cucumbers 406:418 sea cucumbers 406:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 0 36 theme model 30:34 arg1 compounds 36:44 model compounds 30:44 model compounds related to fucosylated chondroitin sulfates 30:88 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 1 37 theme model 316:320 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 37 theme model 316:320 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 37 theme model 316:320 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 0 38 theme GalNAc 112:117 arg1 derivatives 119:129 Fuc(1 → 6)GalNAc derivatives 102:129 Fuc(1 → 6)GalNAc derivatives 102:129 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 3 39 theme β-isomeric 822:831 arg1 disaccharides 833:845 the corresponding α- and β-isomeric disaccharides 797:845 the corresponding α- and β-isomeric disaccharides 797:845 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 4 40 theme Selective 1002:1010 arg1 removal 1012:1018 Selective removal 1002:1018 Selective removal of orthogonal O-protections 1002:1046 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 1 41 theme O-sulfated 242:251 arg1 2-acetamido-2-deoxy-β-D-galactopyranosides 177:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides 132:218 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 41 theme O-sulfated 242:251 arg1 compounds 322:330 model compounds 316:330 model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers 316:418 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 41 theme O-sulfated 242:251 arg1 derivatives 284:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 their selectively O-sulfated (both in GalNAc and Fuc units) derivatives 224:294 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 2 42 theme Per-O-acetylated 421:436 arg1 precursor 496:504 a key precursor 490:504 a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products 490:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 2 42 theme Per-O-acetylated 421:436 arg1 2-deoxy-2-N-phthalimido-D-glucopyranose 438:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 5 43 theme computer 1352:1359 arg1 analysis 1381:1388 computer assisted structural analysis 1352:1388 computer assisted structural analysis of natural FCS 1352:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 1 44 dep both 254:257 arg1 GalNAc 262:267 GalNAc 262:267 GalNAc 262:267 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 5 45 theme Further 1115:1121 arg1 removal 1123:1129 Further removal 1115:1129 Further removal of blocking groups 1115:1148 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 0 46 theme related 46:52 arg1 compounds 36:44 model compounds 30:44 model compounds related to fucosylated chondroitin sulfates 30:88 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 5 47 theme assisted 1361:1368 arg1 analysis 1381:1388 computer assisted structural analysis 1352:1388 computer assisted structural analysis of natural FCS 1352:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 3 48 theme α- 815:816 arg1 disaccharides 833:845 the corresponding α- and β-isomeric disaccharides 797:845 the corresponding α- and β-isomeric disaccharides 797:845 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 5 49 theme NMR 1226:1228 arg1 spectroscopy 1230:1241 13C NMR spectroscopy 1222:1241 13C NMR spectroscopy 1222:1241 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 5 50 theme α-L-fucosylation 1305:1320 arg1 effects 1278:1284 the spectral effects 1265:1284 the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS 1265:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 0 51 theme compounds 36:44 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis 0:8 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 0 51 theme compounds 36:44 arg1 analysis 18:25 NMR analysis 14:25 NMR analysis 14:25 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 5 52 theme structural 1370:1379 arg1 analysis 1381:1388 computer assisted structural analysis 1352:1388 computer assisted structural analysis of natural FCS 1352:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 4 53 theme GalNAc 1093:1098 arg1 units 1108:1112 the GalNAc and Fuc units 1089:1112 units 1108:1112 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 4 54 from units 1108:1112 arg1 O-sulfation 1069:1079 subsequent O-sulfation 1058:1079 subsequent O-sulfation both at the GalNAc and Fuc units 1058:1112 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 3 55 theme TMSOTf 768:773 arg1 presence 756:763 the presence 752:763 the presence of TMSOTf 752:773 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 2 56 used used 482:485 arg2 2-deoxy-2-N-phthalimido-D-glucopyranose 438:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose 421:476 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 2 56 used used 482:485 arg2 precursor 496:504 a key precursor 490:504 a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products 490:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 3 57 theme corresponding 801:813 arg1 disaccharides 833:845 the corresponding α- and β-isomeric disaccharides 797:845 the corresponding α- and β-isomeric disaccharides 797:845 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 58 from trichloracetimidate 729:747 arg1 presence 756:763 the presence 752:763 the presence of TMSOTf 752:773 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 5 59 theme analysis 1381:1388 arg1 development 1337:1347 the development 1333:1347 the development of computer assisted structural analysis of natural FCS 1333:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 1 60 dep O-sulfated 242:251 arg1 units 277:281 both in GalNAc and Fuc units 254:281 both in GalNAc and Fuc units 254:281 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 5 61 theme 13C 1222:1224 arg1 spectroscopy 1230:1241 13C NMR spectroscopy 1222:1241 13C NMR spectroscopy 1222:1241 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 3 62 theme disaccharide 976:987 arg1 formation 944:952 the formation 940:952 the formation of desired α-isomeric disaccharide 940:987 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 4 63 theme Fuc 1104:1106 arg1 units 1108:1112 the GalNAc and Fuc units 1089:1112 units 1108:1112 Selective removal of orthogonal O-protections permitted subsequent O-sulfation both at the GalNAc and Fuc units. 27951443 3 64 theme disaccharides 833:845 arg1 mixture 786:792 a 1:1 mixture 780:792 a 1:1 mixture of the corresponding α- and β-isomeric disaccharides 780:845 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 5 65 theme spectral 1269:1276 arg1 effects 1278:1284 the spectral effects 1265:1284 the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS 1265:1403 Further removal of blocking groups yielded the target products which were systematically studied by 1H and 13C NMR spectroscopy in order to determine the spectral effects of O-sulfation and α-L-fucosylation needed for the development of computer assisted structural analysis of natural FCS. 27951443 0 66 theme fucosylated 57:67 arg1 sulfates 81:88 fucosylated chondroitin sulfates 57:88 fucosylated chondroitin sulfates 57:88 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 1 67 dep units 277:281 arg1 both 254:257 both 254:257 both 254:257 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 1 67 dep units 277:281 arg1 Fuc 273:275 Fuc 273:275 Fuc 273:275 Unsubstituted and 6-O-α-L-fucosylated propyl 2-acetamido-2-deoxy-β-D-galactopyranosides and their selectively O-sulfated (both in GalNAc and Fuc units) derivatives were synthesized as model compounds representing the fragments of fucosylated chondroitin sulfates (FCS) from sea cucumbers. 27951443 3 68 theme 1:1 782:784 arg1 mixture 786:792 a 1:1 mixture 780:792 a 1:1 mixture of the corresponding α- and β-isomeric disaccharides 780:845 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 69 theme desired 957:963 arg1 disaccharide 976:987 desired α-isomeric disaccharide 957:987 desired α-isomeric disaccharide 957:987 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 2 70 theme products 584:591 arg1 preparation 514:524 the preparation 510:524 the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products 510:591 Per-O-acetylated 2-deoxy-2-N-phthalimido-D-glucopyranose was used as a key precursor for the preparation of all 2-acetamido-2-deoxy-D-galactopyranoside containing products. 27951443 0 71 dep Synthesis 0:8 arg1 GalNAc 91:96 GalNAc 91:96 GalNAc 91:96 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 0 71 dep Synthesis 0:8 arg1 derivatives 119:129 Fuc(1 → 6)GalNAc derivatives 102:129 Fuc(1 → 6)GalNAc derivatives 102:129 Synthesis and NMR analysis of model compounds related to fucosylated chondroitin sulfates: GalNAc and Fuc(1 → 6)GalNAc derivatives. 27951443 3 72 theme related 878:884 arg1 donor 902:906 structurally related fucosyl bromide donor 865:906 structurally related fucosyl bromide donor 865:906 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 73 with use 858:860 arg1 promotion 913:921 promotion 913:921 promotion by Bu4NBr 913:931 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 27951443 3 74 theme α-isomeric 965:974 arg1 disaccharide 976:987 desired α-isomeric disaccharide 957:987 desired α-isomeric disaccharide 957:987 Attempts at 6-O-glycosylation of propyl 3-O-benzoyl-2-deoxy-2-N-phthalimido-D-galactoside by 2-O-benzyl-3,4-di-O-chloracetyl-L-fucosyl trichloracetimidate in the presence of TMSOTf gave a 1:1 mixture of the corresponding α- and β-isomeric disaccharides, while the use of structurally related fucosyl bromide donor with promotion by Bu4NBr led to the formation of desired α-isomeric disaccharide exclusively. 28193843 0 0 theme New 79:81 arg1 Model 95:99 a New Mechanistic Model 77:99 a New Mechanistic Model 77:99 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 6 1 theme catalytic 881:889 arg1 site 891:894 the catalytic site 877:894 the catalytic site 877:894 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 11 2 theme binding 1700:1706 arg1 model 1708:1712 a substrate binding model 1688:1712 a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE 1688:1796 Based on these structures, we propose a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE. 28193843 8 3 theme A. 1152:1153 arg1 analysis 1180:1187 A. Site-directed mutational analysis 1152:1187 A. Site-directed mutational analysis 1152:1187 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 5 4 theme 1-773 724:728 arg1 5-759 715:719 5-759 715:719 5-759 715:719 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 5 4 theme 1-773 724:728 arg1 1-773 724:728 1-773 724:728 1-773 724:728 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 0 5 from Substrate 6:14 arg1 Structure 23:31 the Structure 19:31 the Structure of Cyanobacterial Branching Enzyme 19:66 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 1 6 from formation 138:146 arg1 glycogen 196:203 glycogen 196:203 glycogen 196:203 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 1 6 from formation 138:146 arg1 amylopectin 180:190 amylopectin 180:190 amylopectin 180:190 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 10 7 theme domain 1558:1563 arg1 A 1565:1565 domain A 1558:1565 domain A 1558:1565 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 5 8 theme ligand-free 679:689 arg1 BE1 701:703 ligand-free wild-type BE1 679:703 ligand-free wild-type BE1 (residues 5-759 of 1-773) 679:729 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 5 8 theme ligand-free 679:689 arg1 residues 706:713 residues 5-759 of 1-773 706:728 residues 5-759 of 1-773 706:728 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 11 9 theme glycosylation/deglycosylation 1742:1770 arg1 reactions 1772:1780 glycosylation/deglycosylation reactions 1742:1780 glycosylation/deglycosylation reactions catalyzed by BE 1742:1796 Based on these structures, we propose a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE. 28193843 6 10 theme domain 901:906 arg1 C 908:908 domain C 901:908 domain C 901:908 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 0 11 theme Mechanistic 83:93 arg1 Model 95:99 a New Mechanistic Model 77:99 a New Mechanistic Model 77:99 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 9 12 theme active 1373:1378 arg1 cleft 1385:1389 the active site cleft 1369:1389 the active site cleft 1369:1389 In this structure, maltoheptaose was bound in the active site cleft, allowing us to assign subsites -7 to -1. 28193843 11 13 theme substrate 1690:1698 arg1 model 1708:1712 a substrate binding model 1688:1712 a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE 1688:1796 Based on these structures, we propose a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE. 28193843 8 14 theme Site-directed 1155:1167 arg1 analysis 1180:1187 A. Site-directed mutational analysis 1152:1187 A. Site-directed mutational analysis 1152:1187 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 5 15 theme BE1 701:703 arg1 structure 666:674 the crystal structure 654:674 the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution 654:750 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 6 16 theme module 831:836 arg1 family 838:843 carbohydrate-binding module family 48 810:846 carbohydrate-binding module family 48 (CBM48) 810:854 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 6 16 theme module 831:836 arg1 CBM48 849:853 CBM48 849:853 CBM48 849:853 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 7 17 theme other 975:979 arg1 domains 981:987 the other domains 971:987 the other domains 971:987 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 1 18 dep enzyme 112:117 arg1 BE 120:121 BE 120:121 BE 120:121 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 8 19 theme hydrophobic 1105:1115 arg1 interactions 1117:1128 hydrophobic interactions 1105:1128 hydrophobic interactions with CBM48 and domain 1105:1150 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 1 20 theme α-1,6-glucosidic 151:166 arg1 linkages 168:175 α-1,6-glucosidic linkages 151:175 α-1,6-glucosidic linkages 151:175 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 5 21 theme Å 739:739 arg1 resolution 741:750 1.85 Å resolution 734:750 1.85 Å resolution 734:750 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 5 22 from resolution 741:750 arg1 structure 666:674 the crystal structure 654:674 the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution 654:750 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 7 23 dep β-sandwich 995:1004 arg1 folds 1006:1010 folds 1006:1010 folds 1006:1010 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 3 24 contain have 372:375 arg1 cyanobacteria 358:370 most cyanobacteria 353:370 most cyanobacteria 353:370 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 3 24 contain have 372:375 arg2 one 382:384 one 382:384 one 382:384 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 2 25 theme organism 259:266 arg1 sources 268:274 the organism sources 255:274 the organism sources 255:274 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 1 26 theme linkages 168:175 arg1 formation 138:146 the formation 134:146 the formation of α-1,6-glucosidic linkages in amylopectin and glycogen 134:203 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 10 27 theme oligosaccharide-binding 1449:1471 arg1 sites 1473:1477 seven oligosaccharide-binding sites 1443:1477 seven oligosaccharide-binding sites 1443:1477 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 8 28 with interactions 1117:1128 arg1 domain 1145:1150 domain 1145:1150 domain 1145:1150 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 8 28 with interactions 1117:1128 arg1 CBM48 1135:1139 CBM48 1135:1139 CBM48 1135:1139 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 10 29 theme glucan 1621:1626 arg1 chains 1628:1633 the donor/acceptor glucan chains 1602:1633 the donor/acceptor glucan chains 1602:1633 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 8 30 dep back 1057:1060 arg1 identified 1189:1198 identified 1189:1198 identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution 1189:1320 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 3 31 theme glycoside 410:418 arg1 family 430:435 glycoside hydrolase family 13 410:438 glycoside hydrolase family 13 410:438 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 3 31 theme glycoside 410:418 arg1 sp 452:453 Cyanothece sp 441:453 Cyanothece sp 441:453 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 0 32 theme Bound 0:4 arg1 Substrate 6:14 Bound Substrate 0:14 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme 0:66 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 10 33 theme donor/acceptor 1606:1619 arg1 chains 1628:1633 the donor/acceptor glucan chains 1602:1633 the donor/acceptor glucan chains 1602:1633 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 3 34 theme hydrolase 420:428 arg1 family 430:435 glycoside hydrolase family 13 410:438 glycoside hydrolase family 13 410:438 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 3 34 theme hydrolase 420:428 arg1 sp 452:453 Cyanothece sp 441:453 Cyanothece sp 441:453 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 8 35 theme new 1037:1039 arg1 location 1041:1048 a new location 1035:1048 a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution 1035:1320 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 10 36 theme protein 1502:1508 arg1 surface 1510:1516 the protein surface 1498:1516 the protein surface 1498:1516 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 2 37 theme mechanistic 285:295 arg1 basis 297:301 the mechanistic basis 281:301 the mechanistic basis for these different outcomes 281:330 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 2 37 theme mechanistic 285:295 arg1 unclear 335:341 unclear 335:341 unclear 335:341 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 4 38 theme enzymatic 520:528 arg1 properties 530:539 distinct enzymatic properties 511:539 distinct enzymatic properties 511:539 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 5 39 theme crystal 658:664 arg1 structure 666:674 the crystal structure 654:674 the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution 654:750 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 10 40 theme these 1549:1553 arg1 these 1549:1553 these 1549:1553 these 1549:1553 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 10 40 theme these 1549:1553 arg1 two 1542:1544 two 1542:1544 two 1542:1544 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 1 41 theme Branching 102:110 arg1 enzyme 112:117 Branching enzyme 102:117 Branching enzyme (BE) 102:122 Branching enzyme (BE) catalyzes the formation of α-1,6-glucosidic linkages in amylopectin and glycogen. 28193843 8 42 from back 1057:1060 arg1 location 1041:1048 a new location 1035:1048 a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution 1035:1320 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 10 43 dep entrance 1581:1588 arg1 the 1577:1579 the 1577:1579 the 1577:1579 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 8 44 with bonds 1095:1099 arg1 domain 1145:1150 domain 1145:1150 domain 1145:1150 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 8 44 with bonds 1095:1099 arg1 CBM48 1135:1139 CBM48 1135:1139 CBM48 1135:1139 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 4 45 theme distinct 511:518 arg1 properties 530:539 distinct enzymatic properties 511:539 distinct enzymatic properties 511:539 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 5 46 dep residues 706:713 arg1 5-759 715:719 5-759 715:719 5-759 715:719 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 5 46 dep residues 706:713 arg1 1-773 724:728 1-773 724:728 1-773 724:728 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 6 47 theme domain 800:805 arg1 N 807:807 domain N 800:807 domain N 800:807 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 10 48 theme chains 1628:1633 arg1 exit 1594:1597 exit 1594:1597 exit 1594:1597 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 10 48 theme chains 1628:1633 arg1 entrance 1581:1588 entrance 1581:1588 entrance 1581:1588 Moreover, seven oligosaccharide-binding sites were identified on the protein surface, and we postulated that two of these in domain A served as the entrance and exit of the donor/acceptor glucan chains, respectively. 28193843 11 49 theme reactions 1772:1780 arg1 mechanism 1729:1737 the mechanism 1725:1737 the mechanism of glycosylation/deglycosylation reactions catalyzed by BE 1725:1796 Based on these structures, we propose a substrate binding model explaining the mechanism of glycosylation/deglycosylation reactions catalyzed by BE. 28193843 7 50 theme 8-barrel 948:955 arg1 fold 957:960 a (β/α)8-barrel fold 941:960 a (β/α)8-barrel fold 941:960 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 5 51 theme wild-type 691:699 arg1 BE1 701:703 ligand-free wild-type BE1 679:703 ligand-free wild-type BE1 (residues 5-759 of 1-773) 679:729 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 5 51 theme wild-type 691:699 arg1 residues 706:713 residues 5-759 of 1-773 706:728 residues 5-759 of 1-773 706:728 Here, we report the crystal structure of ligand-free wild-type BE1 (residues 5-759 of 1-773) at 1.85 Å resolution. 28193843 7 52 theme domain 923:928 arg1 A 930:930 The central domain A 911:930 The central domain A 911:930 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 3 53 theme Cyanothece 441:450 arg1 family 430:435 glycoside hydrolase family 13 410:438 glycoside hydrolase family 13 410:438 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 3 53 theme Cyanothece 441:450 arg1 sp 452:453 Cyanothece sp 441:453 Cyanothece sp 441:453 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 28193843 0 54 theme Branching 51:59 arg1 Enzyme 61:66 Cyanobacterial Branching Enzyme 36:66 Cyanobacterial Branching Enzyme 36:66 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 8 55 theme Domain 1013:1018 arg1 N 1020:1020 Domain N 1013:1020 Domain N 1013:1020 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 8 56 theme structure 1277:1285 arg1 determination 1287:1299 structure determination 1277:1299 structure determination 1277:1299 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 6 57 theme carbohydrate-binding 810:829 arg1 family 838:843 carbohydrate-binding module family 48 810:846 carbohydrate-binding module family 48 (CBM48) 810:854 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 6 57 theme carbohydrate-binding 810:829 arg1 CBM48 849:853 CBM48 849:853 CBM48 849:853 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 4 58 contain has 467:469 arg2 BE2 492:494 BE2 492:494 BE2 492:494 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 58 contain has 467:469 arg1 ATCC 456:459 ATCC 51142 456:465 ATCC 51142 456:465 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 58 contain has 467:469 arg2 BE1 487:489 BE1 487:489 BE1 487:489 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 58 contain has 467:469 arg2 BE3 501:503 BE3 501:503 BE3 501:503 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 58 contain has 467:469 arg2 isoforms 477:484 three isoforms 471:484 three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties 471:539 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 0 59 theme Cyanobacterial 36:49 arg1 Enzyme 61:66 Cyanobacterial Branching Enzyme 36:66 Cyanobacterial Branching Enzyme 36:66 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 6 60 theme domain 857:862 arg1 A 864:864 domain A 857:864 domain A containing the catalytic site 857:894 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 7 61 theme β/α 944:946 arg1 fold 957:960 a (β/α)8-barrel fold 941:960 a (β/α)8-barrel fold 941:960 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 2 62 theme reaction 210:217 arg1 variable 232:239 variable 232:239 variable 232:239 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 2 62 theme reaction 210:217 arg1 products 219:226 The reaction products 206:226 The reaction products 206:226 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 9 63 dep subsites 1414:1421 arg1 to 1426:1427 to 1426:1427 to 1426:1427 In this structure, maltoheptaose was bound in the active site cleft, allowing us to assign subsites -7 to -1. 28193843 4 64 dep isoforms 477:484 arg1 BE2 492:494 BE2 492:494 BE2 492:494 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 64 dep isoforms 477:484 arg1 BE3 501:503 BE3 501:503 BE3 501:503 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 64 dep isoforms 477:484 arg1 BE1 487:489 BE1 487:489 BE1 487:489 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 4 64 dep isoforms 477:484 arg1 isoforms 477:484 three isoforms 471:484 three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties 471:539 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 8 65 with maltoheptaose 1228:1240 arg1 affinity 1258:1265 sufficient affinity 1247:1265 sufficient affinity 1247:1265 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 7 66 theme central 915:921 arg1 A 930:930 The central domain A 911:930 The central domain A 911:930 The central domain A displays a (β/α)8-barrel fold, whereas the other domains adopt β-sandwich folds. 28193843 2 67 theme different 313:321 arg1 outcomes 323:330 these different outcomes 307:330 these different outcomes 307:330 The reaction products are variable, depending on the organism sources, and the mechanistic basis for these different outcomes is unclear. 28193843 8 68 theme sufficient 1247:1256 arg1 affinity 1258:1265 sufficient affinity 1247:1265 sufficient affinity 1247:1265 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 8 69 located found 1026:1030 arg2 N 1020:1020 Domain N 1013:1020 Domain N 1013:1020 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 8 69 located found 1026:1030 arg1 location 1041:1048 a new location 1035:1048 a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution 1035:1320 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 4 70 with isoforms 477:484 arg1 properties 530:539 distinct enzymatic properties 511:539 distinct enzymatic properties 511:539 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 8 71 theme mutational 1169:1178 arg1 analysis 1180:1187 A. Site-directed mutational analysis 1152:1187 A. Site-directed mutational analysis 1152:1187 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 4 72 theme unique 604:609 arg1 insights 611:618 unique insights 604:618 unique insights into this system 604:635 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 8 73 theme Å 1309:1309 arg1 resolution 1311:1320 2.30 Å resolution 1304:1320 2.30 Å resolution 1304:1320 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 9 74 theme site 1380:1383 arg1 cleft 1385:1389 the active site cleft 1369:1389 the active site cleft 1369:1389 In this structure, maltoheptaose was bound in the active site cleft, allowing us to assign subsites -7 to -1. 28193843 8 75 theme hydrogen 1086:1093 arg1 bonds 1095:1099 hydrogen bonds 1086:1099 hydrogen bonds 1086:1099 Domain N was found in a new location at the back of the protein, forming hydrogen bonds and hydrophobic interactions with CBM48 and domain A. Site-directed mutational analysis identified a mutant (W610N) that bound maltoheptaose with sufficient affinity to enable structure determination at 2.30 Å resolution. 28193843 4 76 theme enzymes 582:588 arg1 investigations 558:571 investigations 558:571 investigations of these enzymes 558:588 ATCC 51142 has three isoforms (BE1, BE2, and BE3) with distinct enzymatic properties, suggesting that investigations of these enzymes might provide unique insights into this system. 28193843 0 77 theme Enzyme 61:66 arg1 Structure 23:31 the Structure 19:31 the Structure of Cyanobacterial Branching Enzyme 19:66 Bound Substrate in the Structure of Cyanobacterial Branching Enzyme Supports a New Mechanistic Model. 28193843 6 78 contain containing 866:875 arg1 A 864:864 domain A 857:864 domain A containing the catalytic site 857:894 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 6 78 contain containing 866:875 arg2 site 891:894 the catalytic site 877:894 the catalytic site 877:894 The enzyme consists of four domains, including domain N, carbohydrate-binding module family 48 (CBM48), domain A containing the catalytic site, and domain C. 28193843 3 79 theme most 353:356 arg1 cyanobacteria 358:370 most cyanobacteria 353:370 most cyanobacteria 353:370 Although most cyanobacteria have only one BE isoform belonging to glycoside hydrolase family 13, Cyanothece sp. 24848382 6 0 theme putative 1301:1308 arg1 glycans 1319:1325 putative O-linked glycans 1301:1325 putative O-linked glycans 1301:1325 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 4 1 theme Ni-affinity 690:700 arg1 chromatography 702:715 Ni-affinity chromatography 690:715 Ni-affinity chromatography 690:715 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 6 2 theme O-linked 1310:1317 arg1 glycans 1319:1325 putative O-linked glycans 1301:1325 putative O-linked glycans 1301:1325 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 1 3 with cellulase 185:193 arg1 activity 205:212 high activity 200:212 high activity at low temperature 200:231 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 6 4 contain contains 1292:1299 arg2 glycans 1319:1325 putative O-linked glycans 1301:1325 putative O-linked glycans 1301:1325 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 4 contain contains 1292:1299 arg1 Ec-CaCel 1283:1290 Ec-CaCel 1283:1290 Ec-CaCel 1283:1290 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 5 theme 0°C. 1166:1169 arg1 analysis 1198:1205 0°C. Oligosaccharide structural analysis 1166:1205 0°C. Oligosaccharide structural analysis 1166:1205 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 7 6 from mori 1364:1367 arg1 Thermostability 1328:1342 Thermostability 1328:1342 Thermostability of Bm-CaCel from B. mori at 60°C 1328:1375 Thermostability of Bm-CaCel from B. mori at 60°C was higher than that from E. coli, probably due to glycosylation. 24848382 3 7 theme Recombinant 398:408 arg1 rCaCel 417:422 rCaCel 417:422 rCaCel 417:422 Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10°C, but the expression yield was low. 24848382 3 7 theme Recombinant 398:408 arg1 CaCel 410:414 Recombinant CaCel 398:414 Recombinant CaCel (rCaCel) 398:423 Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10°C, but the expression yield was low. 24848382 0 8 from Characterization 0:15 arg1 Bombyx 70:75 Bombyx 70:75 Bombyx 70:75 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 6 9 theme Oligosaccharide 1171:1185 arg1 analysis 1198:1205 0°C. Oligosaccharide structural analysis 1166:1205 0°C. Oligosaccharide structural analysis 1166:1205 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 10 theme temperatures 1096:1107 arg1 range 1073:1077 a broad range 1065:1077 a broad range of pH values and temperatures 1065:1107 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 11 contain contains 1230:1237 arg1 Bm-CaCel 1221:1228 Bm-CaCel 1221:1228 Bm-CaCel 1221:1228 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 11 contain contains 1230:1237 arg2 glycans 1266:1272 O-linked glycans 1257:1272 O-linked glycans 1257:1272 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 11 contain contains 1230:1237 arg2 N- 1250:1251 elaborated N- 1239:1251 elaborated N- 1239:1251 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 12 theme optimal 941:947 arg1 pH 949:950 optimal pH 941:950 optimal pH 941:950 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 5 13 theme specific 732:739 arg1 activity 741:748 specific activity 732:748 specific activity 732:748 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 5 14 dep mg/l 803:806 arg1 mg/l 916:919 0.46 mg/l 911:919 0.46 mg/l 911:919 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 5 14 dep mg/l 803:806 arg1 U/mg 930:933 35.8 U/mg 925:933 35.8 U/mg 925:933 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 5 15 theme CaCel 876:880 arg1 yield 882:886 the CaCel yield 872:886 the CaCel yield obtained from E. coli 872:908 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 2 16 theme expression 378:387 arg1 systems 389:395 Bombyx mori-baculovirus expression systems 354:395 Bombyx mori-baculovirus expression systems 354:395 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 6 17 link O-linked 1310:1317 arg1 glycans 1319:1325 putative O-linked glycans 1301:1325 putative O-linked glycans 1301:1325 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 18 theme values 1085:1090 arg1 range 1073:1077 a broad range 1065:1077 a broad range of pH values and temperatures 1065:1107 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 19 theme activity 1154:1161 arg1 activity 1154:1161 their maximal activity 1140:1161 their maximal activity 1140:1161 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 19 theme activity 1154:1161 arg1 % 1135:1135 more than 30% 1123:1135 more than 30% of their maximal activity 1123:1161 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 20 theme pH 1082:1083 arg1 values 1085:1090 pH values 1082:1090 pH values 1082:1090 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 1 21 theme high 200:203 arg1 activity 205:212 high activity 200:212 high activity at low temperature 200:231 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 7 22 from 60°C 1372:1375 arg1 Thermostability 1328:1342 Thermostability 1328:1342 Thermostability of Bm-CaCel from B. mori at 60°C 1328:1375 Thermostability of Bm-CaCel from B. mori at 60°C was higher than that from E. coli, probably due to glycosylation. 24848382 4 23 theme secretion 566:574 arg1 Bm-CaCel 584:591 Bm-CaCel 584:591 Bm-CaCel 584:591 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 4 23 theme secretion 566:574 arg1 signal 576:581 a silkworm secretion signal 555:581 a silkworm secretion signal (Bm-CaCel) 555:592 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 6 24 theme maximal 1146:1152 arg1 activity 1154:1161 their maximal activity 1140:1161 their maximal activity 1140:1161 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 0 25 from Bombyx 70:75 arg1 Characterization 0:15 Characterization 0:15 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 0:100 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 0 25 from Bombyx 70:75 arg1 endo-β-1,4-glucanase 44:63 Cryptopygus antarcticus endo-β-1,4-glucanase 20:63 Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems 20:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 4 26 theme 90 676:677 arg1 % 678:678 % 678:678 % 678:678 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 4 27 theme silkworm 557:564 arg1 Bm-CaCel 584:591 Bm-CaCel 584:591 Bm-CaCel 584:591 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 4 27 theme silkworm 557:564 arg1 signal 576:581 a silkworm secretion signal 555:581 a silkworm secretion signal (Bm-CaCel) 555:592 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 4 28 with rCaCel 543:548 arg1 Bm-CaCel 584:591 Bm-CaCel 584:591 Bm-CaCel 584:591 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 4 28 with rCaCel 543:548 arg1 signal 576:581 a silkworm secretion signal 555:581 a silkworm secretion signal (Bm-CaCel) 555:592 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 6 29 theme structural 1187:1196 arg1 analysis 1198:1205 0°C. Oligosaccharide structural analysis 1166:1205 0°C. Oligosaccharide structural analysis 1166:1205 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 0 30 theme antarcticus 32:42 arg1 endo-β-1,4-glucanase 44:63 Cryptopygus antarcticus endo-β-1,4-glucanase 20:63 Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems 20:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 7 31 theme Bm-CaCel 1347:1354 arg1 Thermostability 1328:1342 Thermostability 1328:1342 Thermostability of Bm-CaCel from B. mori at 60°C 1328:1375 Thermostability of Bm-CaCel from B. mori at 60°C was higher than that from E. coli, probably due to glycosylation. 24848382 6 32 theme O-linked 1257:1264 arg1 glycans 1266:1272 O-linked glycans 1257:1272 O-linked glycans 1257:1272 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 1 33 from springtail 146:155 arg1 CaCel 124:128 CaCel 124:128 CaCel 124:128 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 1 33 from springtail 146:155 arg1 Endo-β-1,4-glucanase 102:121 Endo-β-1,4-glucanase 102:121 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, 102:232 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 6 34 theme broad 1067:1071 arg1 range 1073:1077 a broad range 1065:1077 a broad range of pH values and temperatures 1065:1107 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 1 35 theme low 217:219 arg1 temperature 221:231 low temperature 217:231 low temperature 217:231 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 0 36 theme Cryptopygus 20:30 arg1 endo-β-1,4-glucanase 44:63 Cryptopygus antarcticus endo-β-1,4-glucanase 20:63 Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems 20:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 2 37 theme CaCel 326:330 arg1 gene 332:335 CaCel gene 326:335 CaCel gene 326:335 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 3 38 theme expression 511:520 arg1 low 532:534 low 532:534 low 532:534 Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10°C, but the expression yield was low. 24848382 3 38 theme expression 511:520 arg1 yield 522:526 the expression yield 507:526 the expression yield 507:526 Recombinant CaCel (rCaCel) has been expressed in Escherichia coli (Ec-CaCel) at temperatures below 10°C, but the expression yield was low. 24848382 6 39 from range 1073:1077 arg1 active 1055:1060 active 1055:1060 active 1055:1060 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 0 40 theme endo-β-1,4-glucanase 44:63 arg1 Characterization 0:15 Characterization 0:15 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 0:100 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 6 41 theme elaborated 1239:1248 arg1 N- 1250:1251 elaborated N- 1239:1251 elaborated N- 1239:1251 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 0 42 dep Bombyx 70:75 arg1 systems 93:99 expression systems 82:99 Bombyx mori expression systems 70:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 0 42 dep Bombyx 70:75 arg1 mori 77:80 Bombyx mori expression systems 70:99 Bombyx mori expression systems 70:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 5 43 dep yield 722:726 arg1 The 718:720 The 718:720 The 718:720 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 1 44 theme Antarctic 136:144 arg1 springtail 146:155 Antarctic springtail 136:155 Antarctic springtail 136:155 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 1 44 theme Antarctic 136:144 arg1 antarcticus 170:180 Cryptopygus antarcticus 158:180 Cryptopygus antarcticus 158:180 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 1 44 theme Antarctic 136:144 arg1 cellulase 185:193 a cellulase 183:193 a cellulase with high activity at low temperature 183:231 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 6 45 dep pH 949:950 arg1 The 937:939 The 937:939 The 937:939 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 46 dep 50°C. 1031:1035 arg1 retained 1114:1121 retained 1114:1121 retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans 1114:1325 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 6 46 dep 50°C. 1031:1035 arg1 active 1055:1060 active 1055:1060 active 1055:1060 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 2 47 theme mori-baculovirus 361:376 arg1 systems 389:395 Bombyx mori-baculovirus expression systems 354:395 Bombyx mori-baculovirus expression systems 354:395 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 2 48 theme active 287:292 arg1 cellulase 294:302 sufficient active cellulase 276:302 sufficient active cellulase 276:302 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 1 49 from temperature 221:231 arg1 activity 205:212 high activity 200:212 high activity at low temperature 200:231 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 2 50 theme Bombyx 354:359 arg1 systems 389:395 Bombyx mori-baculovirus expression systems 354:395 Bombyx mori-baculovirus expression systems 354:395 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 1 51 theme potential 240:248 arg1 use 261:263 potential industrial use 240:263 potential industrial use 240:263 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 2 52 theme sufficient 276:285 arg1 cellulase 294:302 sufficient active cellulase 276:302 sufficient active cellulase 276:302 To obtain sufficient active cellulase for characterization, CaCel gene was expressed in Bombyx mori-baculovirus expression systems. 24848382 1 53 theme Cryptopygus 158:168 arg1 springtail 146:155 Antarctic springtail 136:155 Antarctic springtail 136:155 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 1 53 theme Cryptopygus 158:168 arg1 antarcticus 170:180 Cryptopygus antarcticus 158:180 Cryptopygus antarcticus 158:180 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 6 54 link O-linked 1257:1264 arg1 glycans 1266:1272 O-linked glycans 1257:1272 O-linked glycans 1257:1272 The optimal pH and temperature for the rCaCels purified from E. coli and B. mori were 3.5 and 50°C. Both rCaCels were active at a broad range of pH values and temperatures, and retained more than 30% of their maximal activity at 0°C. Oligosaccharide structural analysis revealed that Bm-CaCel contains elaborated N- and O-linked glycans, whereas Ec-CaCel contains putative O-linked glycans. 24848382 1 55 theme industrial 250:259 arg1 use 261:263 potential industrial use 240:263 potential industrial use 240:263 Endo-β-1,4-glucanase (CaCel) from Antarctic springtail, Cryptopygus antarcticus, a cellulase with high activity at low temperature, shows potential industrial use. 24848382 5 56 theme rCaCel 753:758 arg1 activity 741:748 specific activity 732:748 specific activity 732:748 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 5 56 theme rCaCel 753:758 arg1 yield 722:726 yield 722:726 yield 722:726 The yield and specific activity of rCaCel purified from B. mori were estimated at 31 mg/l and 43.2 U/mg, respectively, which is significantly higher than the CaCel yield obtained from E. coli (0.46 mg/l and 35.8 U/mg). 24848382 4 57 theme pupal 639:643 arg1 hemolymph 645:653 pupal hemolymph 639:653 pupal hemolymph 639:653 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 24848382 0 58 theme expression 82:91 arg1 systems 93:99 expression systems 82:99 Bombyx mori expression systems 70:99 Characterization of Cryptopygus antarcticus endo-β-1,4-glucanase from Bombyx mori expression systems. 24848382 4 59 theme % 678:678 arg1 purity 680:685 90% purity 676:685 90% purity 676:685 Here, rCaCel with a silkworm secretion signal (Bm-CaCel) was successfully expressed and secreted into pupal hemolymph and purified to near 90% purity by Ni-affinity chromatography. 28693729 2 0 theme mass 660:663 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 0 theme mass 660:663 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 8 1 theme clinical 1966:1973 arg1 discovery 1992:2000 clinical glycan biomarker discovery 1966:2000 clinical glycan biomarker discovery 1966:2000 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 3 2 theme SSAP 719:722 arg1 linkage-specificity 689:707 High linkage-specificity 684:707 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), 684:845 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 2 theme SSAP 719:722 arg1 % 713:713 ∼99% 710:713 ∼99% 710:713 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 6 3 theme glycans 1459:1465 arg1 different 1472:1480 different 1472:1480 different 1472:1480 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 6 3 theme glycans 1459:1465 arg1 abundances 1434:1443 the relative abundances 1421:1443 the relative abundances of individual glycans 1421:1465 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 2 4 theme TOF 655:657 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 4 theme TOF 655:657 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 3 5 theme glycans 1024:1030 arg1 limit 992:996 increased detection limit 972:996 increased detection limit (>10-fold) of derivatized glycans 972:1030 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 5 theme glycans 1024:1030 arg1 >10-fold 999:1006 >10-fold 999:1006 >10-fold 999:1006 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 6 theme common 745:750 arg1 forms 752:756 the two most common forms 732:756 both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), 727:845 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 6 7 theme RA 1540:1541 arg1 ratios 1559:1564 the RA patient/control ratios 1536:1564 meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans 1526:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 6 8 gly α2,6-sialylated 1584:1598 arg1 glycans 1600:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 3 9 theme sensitive 1048:1056 arg1 glycomics 1058:1066 sensitive glycomics 1048:1066 sensitive glycomics 1048:1066 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 10 theme acid 768:771 arg1 forms 752:756 the two most common forms 732:756 both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), 727:845 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 2 11 theme desorption/ionization 610:630 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 11 theme desorption/ionization 610:630 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 4 12 theme reaction 1106:1113 arg1 efficiency 1115:1124 reaction efficiency 1106:1124 reaction efficiency 1106:1124 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 1 13 attach derived 336:342 arg2 structures 325:334 diverse glycan structures 310:334 diverse glycan structures derived from biological samples 310:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 1 13 attach derived 336:342 arg1 samples 360:366 biological samples 349:366 biological samples 349:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 6 14 theme positive 1504:1511 arg1 samples 1513:1519 RA negative and RA positive samples 1485:1519 RA negative and RA positive samples 1485:1519 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 2 15 theme matrix-assisted 588:602 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 15 theme matrix-assisted 588:602 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 8 16 theme therapeutic 1941:1951 arg1 proteins 1953:1960 therapeutic proteins 1941:1960 therapeutic proteins 1941:1960 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 6 17 theme glycans 1600:1606 arg1 meanwhile 1526:1534 meanwhile the RA patient/control ratios 1526:1564 meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans 1526:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 2 18 theme time-of-fight 640:652 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 18 theme time-of-fight 640:652 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 3 19 theme N-acetylneuraminic 774:791 arg1 acid 768:771 sialic acid 761:771 sialic acid 761:771 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 19 theme N-acetylneuraminic 774:791 arg1 acid 793:796 N-acetylneuraminic acid 774:796 N-acetylneuraminic acid (Neu5Ac) 774:805 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 19 theme N-acetylneuraminic 774:791 arg1 Neu5Ac 799:804 Neu5Ac 799:804 Neu5Ac 799:804 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 0 20 theme cellular 156:163 arg1 processes 165:173 cellular processes 156:173 cellular processes 156:173 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 2 21 theme MALDI 633:637 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 21 theme MALDI 633:637 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 3 22 theme N-glycolylneuraminic 811:830 arg1 acid 768:771 sialic acid 761:771 sialic acid 761:771 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 22 theme N-glycolylneuraminic 811:830 arg1 Neu5Gc 838:843 Neu5Gc 838:843 Neu5Gc 838:843 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 22 theme N-glycolylneuraminic 811:830 arg1 acid 832:835 N-glycolylneuraminic acid 811:835 N-glycolylneuraminic acid (Neu5Gc) 811:844 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 8 23 theme glycan 1975:1980 arg1 discovery 1992:2000 clinical glycan biomarker discovery 1966:2000 clinical glycan biomarker discovery 1966:2000 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 6 24 theme sialylation 1659:1669 arg1 increase 1647:1654 the increase 1643:1654 the increase of sialylation 1643:1669 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 0 25 theme protein 70:76 arg1 glycosylation 78:90 protein glycosylation 70:90 protein glycosylation 70:90 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 1 26 theme biological 349:358 arg1 samples 360:366 biological samples 349:366 biological samples 349:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 8 27 theme SSAP 1795:1798 arg1 derivatization 1800:1813 SSAP derivatization 1795:1813 SSAP derivatization combined with MALDI-TOF MS 1795:1840 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 3 28 theme α2,6-linked 892:902 arg1 acids 911:915 α2,3- and α2,6-linked sialic acids 882:915 α2,3- and α2,6-linked sialic acids 882:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 4 29 theme sample 1165:1170 arg1 <2 h 1185:1188 <2 h 1185:1188 <2 h 1185:1188 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 4 29 theme sample 1165:1170 arg1 preparation 1172:1182 rapid sample preparation 1159:1182 rapid sample preparation (<2 h) 1159:1189 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 3 30 theme α2,3- 882:886 arg1 acids 911:915 α2,3- and α2,6-linked sialic acids 882:915 α2,3- and α2,6-linked sialic acids 882:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 2 31 theme glycosylation 571:583 arg1 profiling 558:566 the improved profiling 545:566 the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 545:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 4 32 theme glycans 1222:1228 arg1 analysis 1210:1217 robust analysis 1203:1217 robust analysis of glycans 1203:1228 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 5 33 theme SSAP 1263:1266 arg1 strategy 1268:1275 The SSAP strategy 1259:1275 The SSAP strategy 1259:1275 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 0 34 theme MS. 105:107 arg1 glycosylation 117:129 MALDI-TOF MS. Protein glycosylation 95:129 MALDI-TOF MS. Protein glycosylation 95:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 8 35 theme MALDI-TOF 1829:1837 arg1 MS 1839:1840 MALDI-TOF MS 1829:1840 MALDI-TOF MS 1829:1840 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 7 36 theme glycosylation 1694:1706 arg1 changes 1708:1714 the glycosylation changes 1690:1714 the glycosylation changes occurred in human serum protein 1690:1746 Those findings of the glycosylation changes occurred in human serum protein may contribute to the diagnosis of RA. 28693729 2 37 theme derivatization 408:421 arg1 strategy 423:430 a two-step derivatization strategy 397:430 a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 397:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 3 38 theme acids 911:915 arg1 discrimination 864:877 direct discrimination 857:877 direct discrimination of α2,3- and α2,6-linked sialic acids 857:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 5 39 theme serum 1347:1351 arg1 glycosylation 1324:1336 the protein glycosylation 1312:1336 the protein glycosylation of human serum associated with rheumatoid arthritis (RA) 1312:1393 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 0 40 theme Sialic 0:5 arg1 acid 7:10 Sialic acid 0:10 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation 0:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 1 41 theme in-depth 281:288 arg1 characterization 290:305 rapid, sensitive and in-depth characterization 260:305 rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples 260:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 0 42 from disease 202:208 arg1 mammals 213:219 mammals 213:219 mammals 213:219 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 2 43 theme sialic 456:461 arg1 SSAP 484:487 SSAP 484:487 SSAP 484:487 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 43 theme sialic 456:461 arg1 permethylation 468:481 linkage-specific sialic acid permethylation 439:481 linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 439:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 0 44 theme linkage-specific 12:27 arg1 permethylation 29:42 Sialic acid linkage-specific permethylation 0:42 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation 0:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 6 45 from samples 1513:1519 arg1 different 1472:1480 different 1472:1480 different 1472:1480 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 6 45 from samples 1513:1519 arg1 abundances 1434:1443 the relative abundances 1421:1443 the relative abundances of individual glycans 1421:1465 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 0 46 from development 186:196 arg1 mammals 213:219 mammals 213:219 mammals 213:219 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 5 47 theme protein 1316:1322 arg1 glycosylation 1324:1336 the protein glycosylation 1312:1336 the protein glycosylation of human serum associated with rheumatoid arthritis (RA) 1312:1393 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 1 48 theme glycan 318:323 arg1 structures 325:334 diverse glycan structures 310:334 diverse glycan structures derived from biological samples 310:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 3 49 theme High 684:687 arg1 linkage-specificity 689:707 High linkage-specificity 684:707 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), 684:845 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 49 theme High 684:687 arg1 % 713:713 ∼99% 710:713 ∼99% 710:713 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 4 50 theme good 1083:1086 arg1 compatibility 1088:1100 good compatibility 1083:1100 good compatibility 1083:1100 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 6 51 theme negative 1488:1495 arg1 samples 1513:1519 RA negative and RA positive samples 1485:1519 RA negative and RA positive samples 1485:1519 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 7 52 theme human 1728:1732 arg1 protein 1740:1746 human serum protein 1728:1746 human serum protein 1728:1746 Those findings of the glycosylation changes occurred in human serum protein may contribute to the diagnosis of RA. 28693729 3 53 theme detection 982:990 arg1 limit 992:996 increased detection limit 972:996 increased detection limit (>10-fold) of derivatized glycans 972:1030 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 53 theme detection 982:990 arg1 >10-fold 999:1006 >10-fold 999:1006 >10-fold 999:1006 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 5 54 gly glycosylation 1324:1336 arg1 serum 1347:1351 human serum 1341:1351 human serum associated with rheumatoid arthritis (RA) 1341:1393 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 8 55 theme glycomic 1873:1880 arg1 analysis 1882:1889 glycomic analysis 1873:1889 glycomic analysis 1873:1889 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 1 56 theme rapid 260:264 arg1 characterization 290:305 rapid, sensitive and in-depth characterization 260:305 rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples 260:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 3 57 theme derivatized 1012:1022 arg1 glycans 1024:1030 derivatized glycans 1012:1030 derivatized glycans 1012:1030 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 7 58 theme RA 1783:1784 arg1 diagnosis 1770:1778 the diagnosis 1766:1778 the diagnosis of RA 1766:1784 Those findings of the glycosylation changes occurred in human serum protein may contribute to the diagnosis of RA. 28693729 6 59 theme individual 1448:1457 arg1 glycans 1459:1465 individual glycans 1448:1465 individual glycans 1448:1465 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 1 60 theme sensitive 267:275 arg1 characterization 290:305 rapid, sensitive and in-depth characterization 260:305 rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples 260:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 4 61 theme steps 1137:1141 arg1 compatibility 1088:1100 good compatibility 1083:1100 good compatibility 1083:1100 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 4 61 theme steps 1137:1141 arg1 efficiency 1115:1124 reaction efficiency 1106:1124 reaction efficiency 1106:1124 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 7 62 theme serum 1734:1738 arg1 protein 1740:1746 human serum protein 1728:1746 human serum protein 1728:1746 Those findings of the glycosylation changes occurred in human serum protein may contribute to the diagnosis of RA. 28693729 6 63 dep meanwhile 1526:1534 arg1 ratios 1559:1564 the RA patient/control ratios 1536:1564 meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans 1526:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 4 64 theme SSAP 1146:1149 arg1 steps 1137:1141 the two steps 1129:1141 the two steps of SSAP 1129:1149 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 8 65 theme glycosylation 1914:1926 arg1 profiling 1928:1936 glycosylation profiling 1914:1936 glycosylation profiling of therapeutic proteins 1914:1960 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 0 66 theme wide 142:145 arg1 range 147:151 a wide range 140:151 a wide range of cellular processes 140:173 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 2 67 theme laser 604:608 arg1 MS 679:680 MS 679:680 MS 679:680 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 67 theme laser 604:608 arg1 spectrometry 665:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry 588:676 matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 588:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 3 68 theme sialic 761:766 arg1 acid 768:771 sialic acid 761:771 sialic acid 761:771 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 68 theme sialic 761:766 arg1 acid 793:796 N-acetylneuraminic acid 774:796 N-acetylneuraminic acid (Neu5Ac) 774:805 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 68 theme sialic 761:766 arg1 acid 832:835 N-glycolylneuraminic acid 811:835 N-glycolylneuraminic acid (Neu5Gc) 811:844 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 8 69 theme proteins 1953:1960 arg1 profiling 1928:1936 glycosylation profiling 1914:1936 glycosylation profiling of therapeutic proteins 1914:1960 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 8 69 theme proteins 1953:1960 arg1 discovery 1992:2000 clinical glycan biomarker discovery 1966:2000 clinical glycan biomarker discovery 1966:2000 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 3 70 link α2,6-linked 892:902 arg1 acids 911:915 α2,3- and α2,6-linked sialic acids 882:915 α2,3- and α2,6-linked sialic acids 882:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 0 71 theme processes 165:173 arg1 range 147:151 a wide range 140:151 a wide range of cellular processes 140:173 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 6 72 theme α2,6-sialylated 1584:1598 arg1 glycans 1600:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 8 73 theme biomarker 1982:1990 arg1 discovery 1992:2000 clinical glycan biomarker discovery 1966:2000 clinical glycan biomarker discovery 1966:2000 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 6 74 theme α2,3- 1574:1578 arg1 glycans 1600:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 both α2,3- and α2,6-sialylated glycans 1569:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 4 75 theme rapid 1159:1163 arg1 <2 h 1185:1188 <2 h 1185:1188 <2 h 1185:1188 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 4 75 theme rapid 1159:1163 arg1 preparation 1172:1182 rapid sample preparation 1159:1182 rapid sample preparation (<2 h) 1159:1189 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 0 76 theme glycosylation 78:90 arg1 profiling 57:65 improved profiling 48:65 improved profiling of protein glycosylation 48:90 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 3 77 theme direct 857:862 arg1 discrimination 864:877 direct discrimination 857:877 direct discrimination of α2,3- and α2,6-linked sialic acids 857:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 0 78 theme MALDI-TOF 95:103 arg1 glycosylation 117:129 MALDI-TOF MS. Protein glycosylation 95:129 MALDI-TOF MS. Protein glycosylation 95:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 6 79 theme patient/control 1543:1557 arg1 ratios 1559:1564 the RA patient/control ratios 1536:1564 meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans 1526:1606 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 3 80 theme sialic 904:909 arg1 acids 911:915 α2,3- and α2,6-linked sialic acids 882:915 α2,3- and α2,6-linked sialic acids 882:915 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 0 81 theme Protein 109:115 arg1 glycosylation 117:129 MALDI-TOF MS. Protein glycosylation 95:129 MALDI-TOF MS. Protein glycosylation 95:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 4 82 theme high-throughput 1235:1249 arg1 manner 1251:1256 a high-throughput manner 1233:1256 a high-throughput manner 1233:1256 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 7 83 theme changes 1708:1714 arg1 findings 1678:1685 Those findings 1672:1685 Those findings of the glycosylation changes occurred in human serum protein 1672:1746 Those findings of the glycosylation changes occurred in human serum protein may contribute to the diagnosis of RA. 28693729 2 84 theme improved 549:556 arg1 profiling 558:566 the improved profiling 545:566 the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 545:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 4 85 theme robust 1203:1208 arg1 analysis 1210:1217 robust analysis 1203:1217 robust analysis of glycans 1203:1228 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 5 86 theme human 1341:1345 arg1 serum 1347:1351 human serum 1341:1351 human serum associated with rheumatoid arthritis (RA) 1341:1393 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 2 87 theme two-step 399:406 arg1 strategy 423:430 a two-step derivatization strategy 397:430 a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 397:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 0 88 theme acid 7:10 arg1 permethylation 29:42 Sialic acid linkage-specific permethylation 0:42 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation 0:129 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 4 89 dep compatibility 1088:1100 arg1 the 1079:1081 the 1079:1081 the 1079:1081 Moreover, the good compatibility and reaction efficiency of the two steps of SSAP allowed rapid sample preparation (<2 h), benefiting robust analysis of glycans in a high-throughput manner. 28693729 6 90 dep elevate 1618:1624 arg1 accompanied 1626:1636 accompanied 1626:1636 to elevate accompanied with the increase of sialylation 1615:1669 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 6 91 theme relative 1425:1432 arg1 different 1472:1480 different 1472:1480 different 1472:1480 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 6 91 theme relative 1425:1432 arg1 abundances 1434:1443 the relative abundances 1421:1443 the relative abundances of individual glycans 1421:1465 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 8 92 theme unique 1851:1856 arg1 advantages 1858:1867 unique advantages 1851:1867 unique advantages for glycomic analysis 1851:1889 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 2 93 theme acid 463:466 arg1 SSAP 484:487 SSAP 484:487 SSAP 484:487 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 93 theme acid 463:466 arg1 permethylation 468:481 linkage-specific sialic acid permethylation 439:481 linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 439:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 94 theme linkage-specific 439:454 arg1 SSAP 484:487 SSAP 484:487 SSAP 484:487 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 2 94 theme linkage-specific 439:454 arg1 permethylation 468:481 linkage-specific sialic acid permethylation 439:481 linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS) 439:681 In this study, we described a two-step derivatization strategy termed linkage-specific sialic acid permethylation (SSAP) consisting of dimethylamination and permethylation for the improved profiling of glycosylation by matrix-assisted laser desorption/ionization (MALDI) time-of-fight (TOF) mass spectrometry (MS). 28693729 0 95 theme improved 48:55 arg1 profiling 57:65 improved profiling 48:65 improved profiling of protein glycosylation 48:90 Sialic acid linkage-specific permethylation for improved profiling of protein glycosylation by MALDI-TOF MS. Protein glycosylation mediates a wide range of cellular processes, affecting development and disease in mammals. 28693729 1 96 theme diverse 310:316 arg1 structures 325:334 diverse glycan structures 310:334 diverse glycan structures derived from biological samples 310:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 3 97 theme enhanced 938:945 arg1 >10-fold 958:965 >10-fold 958:965 >10-fold 958:965 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 97 theme enhanced 938:945 arg1 intensity 947:955 The enhanced intensity 934:955 The enhanced intensity (>10-fold) 934:966 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 1 98 theme structures 325:334 arg1 characterization 290:305 rapid, sensitive and in-depth characterization 260:305 rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples 260:366 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 3 99 theme increased 972:980 arg1 limit 992:996 increased detection limit 972:996 increased detection limit (>10-fold) of derivatized glycans 972:1030 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 3 99 theme increased 972:980 arg1 >10-fold 999:1006 >10-fold 999:1006 >10-fold 999:1006 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 8 100 from potential 1901:1909 arg1 profiling 1928:1936 glycosylation profiling 1914:1936 glycosylation profiling of therapeutic proteins 1914:1960 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 8 100 from potential 1901:1909 arg1 discovery 1992:2000 clinical glycan biomarker discovery 1966:2000 clinical glycan biomarker discovery 1966:2000 Herein, SSAP derivatization combined with MALDI-TOF MS exhibits unique advantages for glycomic analysis and shows potential in glycosylation profiling of therapeutic proteins and clinical glycan biomarker discovery. 28693729 3 101 dep forms 752:756 arg1 both 727:730 both 727:730 both 727:730 High linkage-specificity (∼99%) of SSAP to both the two most common forms of sialic acid, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), permitted direct discrimination of α2,3- and α2,6-linked sialic acids in MALDI-TOF MS. The enhanced intensity (>10-fold) and increased detection limit (>10-fold) of derivatized glycans were valued for sensitive glycomics. 28693729 1 102 theme glycocodes 239:248 arg1 " 249:249 the "glycocodes" 234:249 the "glycocodes" 234:249 Deciphering the "glycocodes" requires rapid, sensitive and in-depth characterization of diverse glycan structures derived from biological samples. 28693729 6 103 from different 1472:1480 arg1 samples 1513:1519 RA negative and RA positive samples 1485:1519 RA negative and RA positive samples 1485:1519 It was demonstrated that the relative abundances of individual glycans were different in RA negative and RA positive samples, and meanwhile the RA patient/control ratios of both α2,3- and α2,6-sialylated glycans tended to elevate accompanied with the increase of sialylation. 28693729 5 104 theme rheumatoid 1369:1378 arg1 RA 1391:1392 RA 1391:1392 RA 1391:1392 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 28693729 5 104 theme rheumatoid 1369:1378 arg1 arthritis 1380:1388 rheumatoid arthritis 1369:1388 rheumatoid arthritis (RA) 1369:1393 The SSAP strategy was further applied to investigate the protein glycosylation of human serum associated with rheumatoid arthritis (RA). 25900423 7 0 theme cell 1277:1280 arg1 epitopes 1300:1307 cell wall carbohydrate epitopes 1277:1307 cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1277:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 6 1 theme non-transgenic 919:932 arg1 plants 934:939 non-transgenic plants 919:939 non-transgenic plants 919:939 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 7 2 theme wall 1215:1218 arg1 content 1235:1241 cell wall monosaccharide content 1210:1241 cell wall monosaccharide content 1210:1241 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 2 3 theme enzymes 346:352 arg1 impact 312:317 the overall impact 300:317 the overall impact of non-plant glycosylation enzymes in stable transformed plants 300:381 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 1 4 theme expression 208:217 arg1 plants 198:203 plants 198:203 plants 198:203 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 1 4 theme expression 208:217 arg1 systems 219:225 expression systems 208:225 expression systems for therapeutic proteins 208:250 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 7 5 located detected 1383:1390 arg1 plants 1415:1420 (ST)GalT expressing plants 1395:1420 (ST)GalT expressing plants 1395:1420 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 7 5 located detected 1383:1390 arg2 alterations 1244:1254 alterations 1244:1254 alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1244:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 7 6 theme wall 1282:1285 arg1 epitopes 1300:1307 cell wall carbohydrate epitopes 1277:1307 cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1277:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 4 7 theme developmental 675:687 arg1 retardation 689:699 developmental retardation 675:699 developmental retardation 675:699 While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. 25900423 6 8 theme expressing 955:964 arg1 plants 966:971 all (ST)GalT expressing plants 942:971 all (ST)GalT expressing plants 942:971 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 7 9 from alterations 1244:1254 arg1 reactivity 1263:1272 the reactivity 1259:1272 the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1259:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 10 10 theme important 1719:1727 arg1 issues 1729:1734 Such important issues 1714:1734 Such important issues 1714:1734 Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. 25900423 1 11 theme type 144:147 arg1 structures 149:158 human type structures 138:158 human type structures 138:158 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 1 12 theme pathway 122:128 arg1 strategy 176:183 an important strategy 163:183 an important strategy to implement plants as expression systems for therapeutic proteins 163:250 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 1 12 theme pathway 122:128 arg1 Modification 80:91 Modification 80:91 Modification of the plant N-glycosylation pathway towards human type structures 80:158 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 6 13 theme core 1052:1055 arg1 fucose 1057:1062 core fucose 1052:1062 core fucose 1052:1062 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 5 14 dep increased 753:761 arg1 ST 764:765 ST 764:765 ST 764:765 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 8 15 theme hypersensitive 1544:1557 arg1 reactions 1559:1567 hypersensitive reactions 1544:1567 hypersensitive reactions 1544:1567 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 7 16 theme GalT 1399:1402 arg1 plants 1415:1420 (ST)GalT expressing plants 1395:1420 (ST)GalT expressing plants 1395:1420 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 5 17 theme protein 781:787 arg1 levels 789:794 both increased (ST)GalT mRNA and protein levels 748:794 levels 789:794 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 9 18 theme unfavorable 1596:1606 arg1 modifications 1621:1633 unfavorable phenotypical modifications 1596:1633 unfavorable phenotypical modifications 1596:1633 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 6 19 dep depleted 1040:1047 arg1 largely 1032:1038 largely 1032:1038 largely 1032:1038 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 5 20 theme mRNA 772:775 arg1 levels 789:794 both increased (ST)GalT mRNA and protein levels 748:794 levels 789:794 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 7 21 theme nucleotide 1177:1186 arg1 composition 1194:1204 nucleotide sugar composition 1177:1204 nucleotide sugar composition 1177:1204 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 9 22 theme stable 1650:1655 arg1 expression 1667:1676 stable in planta expression 1650:1676 stable in planta expression of non-native glycosyltransferases 1650:1711 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 6 23 dep 40 1086:1087 arg1 to 1083:1084 to 1083:1084 to 1083:1084 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 5 24 theme N-glycosylation 837:851 arg1 profiles 853:860 N-glycosylation profiles 837:860 N-glycosylation profiles of plants with and without the phenotype 837:901 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 1 25 theme N-glycosylation 106:120 arg1 pathway 122:128 the plant N-glycosylation pathway 96:128 the plant N-glycosylation pathway 96:128 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 3 26 theme transgenic 402:411 arg1 lines 413:417 transgenic lines 402:417 transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT) 402:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 26 theme transgenic 402:411 arg1 benthamiana 430:440 Nicotiana benthamiana 420:440 Nicotiana benthamiana 420:440 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 26 theme transgenic 402:411 arg1 thaliana 458:465 Arabidopsis thaliana 446:465 Arabidopsis thaliana 446:465 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 5 27 gly N-glycosylation 837:851 arg1 plants 865:870 plants 865:870 plants with and without the phenotype 865:901 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 10 28 theme stable 1786:1791 arg1 engineering 1800:1810 stable glycan engineering 1786:1810 stable glycan engineering in plants 1786:1820 Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. 25900423 0 29 theme plants 20:25 arg1 Characterization 0:15 Characterization 0:15 Characterization of plants 0:25 Characterization of plants expressing the human β1,4-galactosyltrasferase gene. 25900423 3 30 theme Arabidopsis 446:456 arg1 lines 413:417 transgenic lines 402:417 transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT) 402:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 30 theme Arabidopsis 446:456 arg1 thaliana 458:465 Arabidopsis thaliana 446:465 Arabidopsis thaliana 446:465 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 7 31 theme pectic 1354:1359 arg1 homogalacturonan 1361:1376 pectic homogalacturonan 1354:1376 pectic homogalacturonan 1354:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 6 32 dep structures 1116:1125 arg1 % 1088:1088 % 1088:1088 % 1088:1088 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 9 33 from planta 1660:1665 arg1 stable 1650:1655 stable 1650:1655 stable 1650:1655 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 7 34 theme sugar 1188:1192 arg1 composition 1194:1204 nucleotide sugar composition 1177:1204 nucleotide sugar composition 1177:1204 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 9 35 theme glycosyltransferases 1692:1711 arg1 expression 1667:1676 stable in planta expression 1650:1676 stable in planta expression of non-native glycosyltransferases 1650:1711 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 8 36 theme increased 1474:1482 arg1 levels 1484:1489 increased levels 1474:1489 increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions 1474:1567 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 0 37 theme β1,4-galactosyltrasferase 48:72 arg1 gene 74:77 the human β1,4-galactosyltrasferase gene 38:77 the human β1,4-galactosyltrasferase gene 38:77 Characterization of plants expressing the human β1,4-galactosyltrasferase gene. 25900423 1 38 theme human 138:142 arg1 structures 149:158 human type structures 138:158 human type structures 138:158 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 6 39 dep processed 1021:1029 arg1 depleted 1040:1047 depleted 1040:1047 depleted of core fucose 1040:1062 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 6 40 with N-glycans 1065:1073 arg1 structures 1116:1125 up to 40% terminally galactosylated structures 1080:1125 up to 40% terminally galactosylated structures 1080:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 2 41 theme transformed 364:374 arg1 plants 376:381 stable transformed plants 357:381 stable transformed plants 357:381 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 1 42 theme important 166:174 arg1 strategy 176:183 an important strategy 163:183 an important strategy to implement plants as expression systems for therapeutic proteins 163:250 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 1 42 theme important 166:174 arg1 Modification 80:91 Modification 80:91 Modification of the plant N-glycosylation pathway towards human type structures 80:158 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 4 43 theme severe 623:628 arg1 phenotype 630:638 a severe phenotype 621:638 a severe phenotype associated with stunted growth and developmental retardation 621:699 While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. 25900423 8 44 theme peroxide 1503:1510 arg1 levels 1484:1489 increased levels 1474:1489 increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions 1474:1567 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 6 45 theme ST 947:948 arg1 plants 966:971 all (ST)GalT expressing plants 942:971 all (ST)GalT expressing plants 942:971 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 7 46 theme epitopes 1300:1307 arg1 reactivity 1263:1272 the reactivity 1259:1272 the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1259:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 10 47 from engineering 1800:1810 arg1 plants 1815:1820 plants 1815:1820 plants 1815:1820 Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. 25900423 6 48 theme galactosylated 1101:1114 arg1 structures 1116:1125 up to 40% terminally galactosylated structures 1080:1125 up to 40% terminally galactosylated structures 1080:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 4 49 theme other 599:603 arg1 lines 605:609 other lines 599:609 other lines 599:609 While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. 25900423 7 50 theme transgenic 1134:1143 arg1 plants 1145:1150 transgenic plants 1134:1150 transgenic plants 1134:1150 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 2 51 theme overall 304:310 arg1 impact 312:317 the overall impact 300:317 the overall impact of non-plant glycosylation enzymes in stable transformed plants 300:381 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 10 52 theme Such 1714:1717 arg1 issues 1729:1734 Such important issues 1714:1734 Such important issues 1714:1734 Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. 25900423 10 53 theme glycan 1793:1798 arg1 engineering 1800:1810 stable glycan engineering 1786:1810 stable glycan engineering in plants 1786:1820 Such important issues need to be taken into consideration in respect to stable glycan engineering in plants. 25900423 2 54 theme glycosylation 332:344 arg1 enzymes 346:352 non-plant glycosylation enzymes 322:352 non-plant glycosylation enzymes 322:352 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 5 55 theme plants 865:870 arg1 profiles 853:860 N-glycosylation profiles 837:860 N-glycosylation profiles of plants with and without the phenotype 837:901 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 6 56 theme significant 985:995 arg1 amounts 997:1003 significant amounts 985:1003 significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures 985:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 6 56 theme significant 985:995 arg1 N-glycans 1065:1073 incompletely processed (largely depleted of core fucose) N-glycans 1008:1073 incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures 1008:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 4 57 theme stunted 656:662 arg1 growth 664:669 stunted growth 656:669 stunted growth 656:669 While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. 25900423 5 58 theme increased 753:761 arg1 mRNA 772:775 increased (ST)GalT mRNA 753:775 increased (ST)GalT mRNA 753:775 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 7 59 dep plants 1415:1420 arg1 ST 1396:1397 ST 1396:1397 ST 1396:1397 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 8 60 dep peroxide 1503:1510 arg1 consequence 1529:1539 a consequence 1527:1539 a consequence of hypersensitive reactions 1527:1567 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 6 61 theme GalT 950:953 arg1 plants 966:971 all (ST)GalT expressing plants 942:971 all (ST)GalT expressing plants 942:971 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 5 62 theme phenotype 722:730 arg1 severity 706:713 The severity 702:713 The severity of the phenotype 702:730 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 8 63 theme reactions 1559:1567 arg1 consequence 1529:1539 a consequence 1527:1539 a consequence of hypersensitive reactions 1527:1567 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 3 64 theme human 510:514 arg1 GalT 548:551 (ST)GalT 544:551 (ST)GalT 544:551 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 64 theme human 510:514 arg1 β1,4-galactosyltransferase 516:541 human β1,4-galactosyltransferase 510:541 human β1,4-galactosyltransferase ((ST)GalT) 510:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 2 65 from impact 312:317 arg1 plants 376:381 stable transformed plants 357:381 stable transformed plants 357:381 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 7 66 from differences 1162:1172 arg1 content 1235:1241 cell wall monosaccharide content 1210:1241 cell wall monosaccharide content 1210:1241 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 7 66 from differences 1162:1172 arg1 composition 1194:1204 nucleotide sugar composition 1177:1204 nucleotide sugar composition 1177:1204 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 7 67 theme expressing 1404:1413 arg1 plants 1415:1420 (ST)GalT expressing plants 1395:1420 (ST)GalT expressing plants 1395:1420 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 9 68 theme phenotypical 1608:1619 arg1 modifications 1621:1633 unfavorable phenotypical modifications 1596:1633 unfavorable phenotypical modifications 1596:1633 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 9 69 from stable 1650:1655 arg1 planta 1660:1665 planta 1660:1665 planta 1660:1665 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 3 70 dep lines 413:417 arg1 lines 413:417 transgenic lines 402:417 transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT) 402:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 70 dep lines 413:417 arg1 benthamiana 430:440 Nicotiana benthamiana 420:440 Nicotiana benthamiana 420:440 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 70 dep lines 413:417 arg1 thaliana 458:465 Arabidopsis thaliana 446:465 Arabidopsis thaliana 446:465 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 8 71 with plants 1432:1437 arg1 alterations 1455:1465 phenotypic alterations 1444:1465 phenotypic alterations 1444:1465 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 2 72 theme non-plant 322:330 arg1 enzymes 346:352 non-plant glycosylation enzymes 322:352 non-plant glycosylation enzymes 322:352 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 6 73 theme processed 1021:1029 arg1 N-glycans 1065:1073 incompletely processed (largely depleted of core fucose) N-glycans 1008:1073 incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures 1008:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 2 74 theme stable 357:362 arg1 plants 376:381 stable transformed plants 357:381 stable transformed plants 357:381 Nevertheless, relatively little is known about the overall impact of non-plant glycosylation enzymes in stable transformed plants. 25900423 7 75 theme monosaccharide 1220:1233 arg1 content 1235:1241 cell wall monosaccharide content 1210:1241 cell wall monosaccharide content 1210:1241 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 5 76 theme GalT 767:770 arg1 mRNA 772:775 increased (ST)GalT mRNA 753:775 increased (ST)GalT mRNA 753:775 The severity of the phenotype correlated with both increased (ST)GalT mRNA and protein levels but no differences were observed between N-glycosylation profiles of plants with and without the phenotype. 25900423 7 77 theme cell 1210:1213 arg1 content 1235:1241 cell wall monosaccharide content 1210:1241 cell wall monosaccharide content 1210:1241 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 1 78 theme plant 100:104 arg1 pathway 122:128 the plant N-glycosylation pathway 96:128 the plant N-glycosylation pathway 96:128 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 3 79 theme Nicotiana 420:428 arg1 lines 413:417 transgenic lines 402:417 transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT) 402:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 79 theme Nicotiana 420:428 arg1 benthamiana 430:440 Nicotiana benthamiana 420:440 Nicotiana benthamiana 420:440 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 3 80 dep GalT 548:551 arg1 ST 545:546 ST 545:546 ST 545:546 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 6 81 dep % 1088:1088 arg1 40 1086:1087 40 1086:1087 40 1086:1087 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 0 82 theme human 42:46 arg1 gene 74:77 the human β1,4-galactosyltrasferase gene 38:77 the human β1,4-galactosyltrasferase gene 38:77 Characterization of plants expressing the human β1,4-galactosyltrasferase gene. 25900423 1 83 theme therapeutic 231:241 arg1 proteins 243:250 therapeutic proteins 231:250 therapeutic proteins 231:250 Modification of the plant N-glycosylation pathway towards human type structures is an important strategy to implement plants as expression systems for therapeutic proteins. 25900423 9 84 theme non-native 1681:1690 arg1 glycosyltransferases 1692:1711 non-native glycosyltransferases 1681:1711 non-native glycosyltransferases 1681:1711 Our data demonstrate that unfavorable phenotypical modifications may occur upon stable in planta expression of non-native glycosyltransferases. 25900423 6 85 theme N-glycans 1065:1073 arg1 amounts 997:1003 significant amounts 985:1003 significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures 985:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 6 85 theme N-glycans 1065:1073 arg1 N-glycans 1065:1073 incompletely processed (largely depleted of core fucose) N-glycans 1008:1073 incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures 1008:1125 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 8 86 theme phenotypic 1444:1453 arg1 alterations 1455:1465 phenotypic alterations 1444:1465 phenotypic alterations 1444:1465 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 25900423 3 87 theme modified 490:497 arg1 version 499:505 a modified version 488:505 a modified version of human β1,4-galactosyltransferase ((ST)GalT) 488:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 4 88 theme transgenic 566:575 arg1 plants 577:582 some transgenic plants 561:582 some transgenic plants 561:582 While some transgenic plants grew normally, other lines exhibited a severe phenotype associated with stunted growth and developmental retardation. 25900423 3 89 theme β1,4-galactosyltransferase 516:541 arg1 version 499:505 a modified version 488:505 a modified version of human β1,4-galactosyltransferase ((ST)GalT) 488:552 Here, we analyzed transgenic lines (Nicotiana benthamiana and Arabidopsis thaliana) that stably express a modified version of human β1,4-galactosyltransferase ((ST)GalT). 25900423 7 90 theme carbohydrate 1287:1298 arg1 epitopes 1300:1307 cell wall carbohydrate epitopes 1277:1307 cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan 1277:1376 While transgenic plants showed no differences in nucleotide sugar composition and cell wall monosaccharide content, alterations in the reactivity of cell wall carbohydrate epitopes associated with arabinogalactan-proteins and pectic homogalacturonan were detected in (ST)GalT expressing plants. 25900423 6 91 dep plants 934:939 arg1 contrast 907:914 contrast 907:914 contrast 907:914 In contrast to non-transgenic plants, all (ST)GalT expressing plants synthesized significant amounts of incompletely processed (largely depleted of core fucose) N-glycans with up to 40% terminally galactosylated structures. 25900423 8 92 theme hydrogen 1494:1501 arg1 peroxide 1503:1510 hydrogen peroxide 1494:1510 hydrogen peroxide 1494:1510 Notably, plants with phenotypic alterations showed increased levels of hydrogen peroxide, most probably a consequence of hypersensitive reactions. 27758867 3 0 theme variant 681:687 arg1 structures 689:698 two variant structures 677:698 two variant structures 677:698 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 6 1 theme α-Fuc3N- 1343:1350 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 1 theme α-Fuc3N- 1343:1350 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 2 2 theme mass 351:354 arg1 spectrometry 356:367 mass spectrometry 351:367 mass spectrometry (nano-LC-MS and MS/MS analysis) 351:399 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 3 3 gly glycopeptide 619:630 arg2 glycopeptide 619:630 a LLDGSSTEIR glycopeptide 606:630 a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, 606:699 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 6 4 theme modification 1314:1325 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 4 theme modification 1314:1325 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 2 5 contain carried 453:459 arg1 glycopeptides 434:446 putative glycopeptides 425:446 putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 425:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 5 contain carried 453:459 arg2 variety 463:469 a variety 461:469 a variety 461:469 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 5 contain carried 453:459 arg2 substitutions 484:496 glycoform substitutions 474:496 glycoform substitutions 474:496 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 6 theme glycans 280:286 arg1 characterization 244:259 the structural characterization 229:259 the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 229:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 1 7 theme bacterial 183:191 arg1 species 193:199 many bacterial species 178:199 many bacterial species 178:199 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 4 8 theme taurine-containing 1152:1169 arg1 structure 1192:1200 a taurine-containing peptidylamido-glycan structure 1150:1200 a taurine-containing peptidylamido-glycan structure 1150:1200 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 2 9 theme strains 319:325 arg1 number 295:300 a number 293:300 a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 293:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 10 theme N-acetylhexosamine 543:560 arg1 residue 562:568 an initial N-acetylhexosamine residue 532:568 an initial N-acetylhexosamine residue to Ser or Thr 532:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 6 11 theme -α-Rha3OMe- 1368:1378 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 11 theme -α-Rha3OMe- 1368:1378 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 4 12 theme ion 936:938 arg1 MS/MS 978:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 6 13 theme 1→2 1364:1366 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 13 theme 1→2 1364:1366 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 4 14 theme mass 1060:1063 arg1 measurement 1065:1075 accurate mass measurement 1051:1075 accurate mass measurement 1051:1075 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 3 15 theme carbohydrate 758:769 arg1 moieties 771:778 carbohydrate moieties 758:778 carbohydrate moieties 758:778 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 6 16 theme -α-Rha- 1356:1362 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 16 theme -α-Rha- 1356:1362 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 1 17 theme flagellins 134:143 arg1 property 166:173 a well recognized property 148:173 a well recognized property of many bacterial species 148:199 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 1 17 theme flagellins 134:143 arg1 Glycosylation 117:129 Glycosylation 117:129 Glycosylation of flagellins 117:143 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 2 18 theme flagellar 270:278 arg1 glycans 280:286 novel flagellar glycans 264:286 novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 264:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 3 19 contain contained 732:740 arg1 glycopeptide 719:730 the glycopeptide 715:730 the glycopeptide 715:730 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 3 19 contain contained 732:740 arg2 entity 800:805 a novel structural entity 781:805 a novel structural entity 781:805 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 6 20 theme 1→3 1352:1354 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 20 theme 1→3 1352:1354 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 3 21 theme glycopeptide 619:630 arg1 analysis 594:601 Detailed analysis 585:601 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, 585:699 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 2 22 from number 295:300 arg1 glycans 280:286 novel flagellar glycans 264:286 novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 264:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 22 from number 295:300 arg1 characterization 244:259 the structural characterization 229:259 the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 229:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 4 23 theme structure 1192:1200 arg1 suggestion 1136:1145 the suggestion 1132:1145 the suggestion of a taurine-containing peptidylamido-glycan structure 1132:1200 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 3 24 theme novel 783:787 arg1 entity 800:805 a novel structural entity 781:805 a novel structural entity 781:805 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 1 25 theme recognized 155:164 arg1 property 166:173 a well recognized property 148:173 a well recognized property of many bacterial species 148:199 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 1 25 theme recognized 155:164 arg1 Glycosylation 117:129 Glycosylation 117:129 Glycosylation of flagellins 117:143 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 7 26 theme moiety 1569:1574 arg1 This 1492:1495 This 1492:1495 This 1492:1495 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 7 26 theme moiety 1569:1574 arg1 report 1510:1515 the first report 1500:1515 the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1500:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 2 27 theme substitutions 484:496 arg1 variety 463:469 a variety 461:469 a variety 461:469 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 27 theme substitutions 484:496 arg1 substitutions 484:496 glycoform substitutions 474:496 glycoform substitutions 474:496 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 6 28 theme 1→ 1395:1396 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 28 theme 1→ 1395:1396 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 7 29 theme sulfonated 1537:1546 arg1 moiety 1569:1574 a novel O-linked sulfonated peptidylamido-glycan moiety 1520:1574 a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1520:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 4 30 theme activated 954:962 arg1 MS/MS 978:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 2 31 theme novel 264:268 arg1 glycans 280:286 novel flagellar glycans 264:286 novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 264:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 32 dep spectrometry 356:367 arg1 analysis 391:398 MS/MS analysis 385:398 MS/MS analysis 385:398 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 32 dep spectrometry 356:367 arg1 nano-LC-MS 370:379 nano-LC-MS 370:379 nano-LC-MS 370:379 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 0 33 theme Sulfonated 83:92 arg1 Peptidylamido-glycans 94:114 Novel Sulfonated Peptidylamido-glycans 77:114 Novel Sulfonated Peptidylamido-glycans 77:114 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 7 34 theme novel 1522:1526 arg1 moiety 1569:1574 a novel O-linked sulfonated peptidylamido-glycan moiety 1520:1574 a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1520:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 6 35 theme 1→3 1380:1382 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 35 theme 1→3 1380:1382 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 0 36 theme Hypervirulent 24:36 arg1 difficile 50:58 Hypervirulent Clostridium difficile 24:58 Hypervirulent Clostridium difficile 24:58 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 5 37 theme flagellin 1228:1236 arg1 glycopeptides 1238:1250 flagellin glycopeptides 1228:1250 flagellin glycopeptides 1228:1250 Finally, NMR analysis of flagellin glycopeptides provided complementary information. 27758867 5 38 gly glycopeptides 1238:1250 arg2 glycopeptides 1238:1250 flagellin glycopeptides 1228:1250 flagellin glycopeptides 1228:1250 Finally, NMR analysis of flagellin glycopeptides provided complementary information. 27758867 2 39 theme MS/MS 385:389 arg1 analysis 391:398 MS/MS analysis 385:398 MS/MS analysis 385:398 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 5 40 theme NMR 1212:1214 arg1 analysis 1216:1223 NMR analysis 1212:1223 NMR analysis of flagellin glycopeptides 1212:1250 Finally, NMR analysis of flagellin glycopeptides provided complementary information. 27758867 2 41 theme putative 425:432 arg1 glycopeptides 434:446 putative glycopeptides 425:446 putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 425:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 3 42 gly glycopeptide 719:730 arg2 glycopeptide 719:730 the glycopeptide 715:730 the glycopeptide 715:730 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 2 43 used used 346:349 arg2 We 343:344 We 343:344 We 343:344 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 0 44 theme B 9:9 arg1 Flagellin 11:19 The Type B Flagellin 0:19 The Type B Flagellin of Hypervirulent Clostridium difficile 0:58 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 6 45 theme novel 1411:1415 arg1 moiety 1425:1430 the novel capping moiety 1407:1430 the novel capping moiety 1407:1430 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 4 46 theme electrospray-MS 821:835 arg1 strategies 837:846 electrospray-MS strategies 821:846 electrospray-MS strategies using Q-TOF technology 821:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 0 47 theme Clostridium 38:48 arg1 difficile 50:58 Hypervirulent Clostridium difficile 24:58 Hypervirulent Clostridium difficile 24:58 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 4 48 theme atomic 1100:1105 arg1 compositions 1107:1118 atomic compositions 1100:1118 atomic compositions 1100:1118 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 2 49 theme difficile 333:341 arg1 strains 319:325 hypervirulent strains 305:325 hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 305:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 3 50 theme structural 789:798 arg1 entity 800:805 a novel structural entity 781:805 a novel structural entity 781:805 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 4 51 theme unique 1000:1005 arg1 patterns 1021:1028 unique fragmentation patterns 1000:1028 unique fragmentation patterns 1000:1028 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 5 52 theme glycopeptides 1238:1250 arg1 analysis 1216:1223 NMR analysis 1212:1223 NMR analysis of flagellin glycopeptides 1212:1250 Finally, NMR analysis of flagellin glycopeptides provided complementary information. 27758867 2 53 theme glycopeptides 434:446 arg1 number 415:420 a number 413:420 a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 413:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 5 54 theme complementary 1261:1273 arg1 information 1275:1285 complementary information 1261:1285 complementary information 1261:1285 Finally, NMR analysis of flagellin glycopeptides provided complementary information. 27758867 1 55 theme many 178:181 arg1 species 193:199 many bacterial species 178:199 many bacterial species 178:199 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 4 56 theme strategies 837:846 arg1 variety 810:816 A variety 808:816 A variety of electrospray-MS strategies using Q-TOF technology 808:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 4 56 theme strategies 837:846 arg1 strategies 837:846 electrospray-MS strategies 821:846 electrospray-MS strategies using Q-TOF technology 821:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 4 57 theme peptidylamido-glycan 1171:1190 arg1 structure 1192:1200 a taurine-containing peptidylamido-glycan structure 1150:1200 a taurine-containing peptidylamido-glycan structure 1150:1200 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 1 58 theme species 193:199 arg1 property 166:173 a well recognized property 148:173 a well recognized property of many bacterial species 148:199 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 1 58 theme species 193:199 arg1 Glycosylation 117:129 Glycosylation 117:129 Glycosylation of flagellins 117:143 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 4 59 theme Q-TOF 854:858 arg1 technology 860:869 Q-TOF technology 854:869 Q-TOF technology 854:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 3 60 contain carried 669:675 arg1 digestion 652:660 tryptic digestion 644:660 tryptic digestion 644:660 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 3 60 contain carried 669:675 arg2 structures 689:698 two variant structures 677:698 two variant structures 677:698 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 2 61 theme hypervirulent 305:317 arg1 strains 319:325 hypervirulent strains 305:325 hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 305:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 4 62 theme positive 914:921 arg1 MS/MS 978:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 2 63 theme structural 233:242 arg1 characterization 244:259 the structural characterization 229:259 the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 229:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 3 64 theme Detailed 585:592 arg1 analysis 594:601 Detailed analysis 585:601 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, 585:699 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 4 65 theme negative 927:934 arg1 MS/MS 978:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 6 66 theme glycan 1292:1297 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 66 theme glycan 1292:1297 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 7 67 theme flagellin 1589:1597 arg1 protein 1599:1605 a flagellin protein 1587:1605 a flagellin protein 1587:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 3 68 theme LLDGSSTEIR 608:617 arg1 glycopeptide 619:630 a LLDGSSTEIR glycopeptide 606:630 a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, 606:699 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 2 69 dep strains 319:325 arg1 used 346:349 used 346:349 used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 346:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 7 70 theme peptidylamido-glycan 1548:1567 arg1 moiety 1569:1574 a novel O-linked sulfonated peptidylamido-glycan moiety 1520:1574 a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1520:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 6 71 theme -β-GlcNAc- 1384:1393 arg1 Ser 1398:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser 1343:1400 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 6 71 theme -β-GlcNAc- 1384:1393 arg1 portion 1299:1305 The glycan portion 1288:1305 The glycan portion of the modification 1288:1325 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 0 72 theme Novel 77:81 arg1 Peptidylamido-glycans 94:114 Novel Sulfonated Peptidylamido-glycans 77:114 Novel Sulfonated Peptidylamido-glycans 77:114 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 7 73 theme O-linked 1528:1535 arg1 moiety 1569:1574 a novel O-linked sulfonated peptidylamido-glycan moiety 1520:1574 a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1520:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 2 74 theme glycoform 474:482 arg1 substitutions 484:496 glycoform substitutions 474:496 glycoform substitutions 474:496 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 7 75 link O-linked 1528:1535 arg1 moiety 1569:1574 a novel O-linked sulfonated peptidylamido-glycan moiety 1520:1574 a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1520:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 1 76 gly Glycosylation 117:129 arg1 flagellins 134:143 flagellins 134:143 flagellins 134:143 Glycosylation of flagellins is a well recognized property of many bacterial species. 27758867 3 77 dep moieties 771:778 arg1 addition 746:753 addition 746:753 addition 746:753 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 2 78 theme initial 535:541 arg1 residue 562:568 an initial N-acetylhexosamine residue 532:568 an initial N-acetylhexosamine residue to Ser or Thr 532:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 4 79 theme decomposition 964:976 arg1 MS/MS 978:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 positive and negative ion collisionally activated decomposition MS/MS 914:982 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 0 80 theme Type 4:7 arg1 Flagellin 11:19 The Type B Flagellin 0:19 The Type B Flagellin of Hypervirulent Clostridium difficile 0:58 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 4 81 theme high 1035:1038 arg1 resolution 1040:1049 high resolution 1035:1049 high resolution accurate mass measurement to allow derivation of atomic compositions 1035:1118 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 2 82 from characterization 244:259 arg1 number 295:300 a number 293:300 a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 293:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 4 83 theme accurate 1051:1058 arg1 measurement 1065:1075 accurate mass measurement 1051:1075 accurate mass measurement 1051:1075 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 3 84 theme tryptic 644:650 arg1 digestion 652:660 tryptic digestion 644:660 tryptic digestion 644:660 Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity. 27758867 6 85 theme capping 1417:1423 arg1 moiety 1425:1430 the novel capping moiety 1407:1430 the novel capping moiety 1407:1430 The glycan portion of the modification was assigned as α-Fuc3N-(1→3)-α-Rha-(1→2)-α-Rha3OMe-(1→3)-β-GlcNAc-(1→)Ser, and the novel capping moiety was shown to be comprised of taurine, alanine, and glycine. 27758867 4 86 used used 876:879 arg2 strategies 837:846 electrospray-MS strategies 821:846 electrospray-MS strategies using Q-TOF technology 821:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 4 86 used used 876:879 arg2 variety 810:816 A variety 808:816 A variety of electrospray-MS strategies using Q-TOF technology 808:869 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 4 87 theme fragmentation 1007:1019 arg1 patterns 1021:1028 unique fragmentation patterns 1000:1028 unique fragmentation patterns 1000:1028 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 27758867 0 88 theme difficile 50:58 arg1 Flagellin 11:19 The Type B Flagellin 0:19 The Type B Flagellin of Hypervirulent Clostridium difficile 0:58 The Type B Flagellin of Hypervirulent Clostridium difficile Is Modified with Novel Sulfonated Peptidylamido-glycans. 27758867 7 89 theme first 1504:1508 arg1 This 1492:1495 This 1492:1495 This 1492:1495 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 7 89 theme first 1504:1508 arg1 report 1510:1515 the first report 1500:1515 the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein 1500:1605 This is the first report of a novel O-linked sulfonated peptidylamido-glycan moiety decorating a flagellin protein. 27758867 2 90 gly glycopeptides 434:446 arg2 glycopeptides 434:446 putative glycopeptides 425:446 putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr 425:582 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 2 91 theme C. 330:331 arg1 difficile 333:341 C. difficile 330:341 C. difficile 330:341 In this study, we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile We used mass spectrometry (nano-LC-MS and MS/MS analysis) to identify a number of putative glycopeptides that carried a variety of glycoform substitutions, each of which was linked through an initial N-acetylhexosamine residue to Ser or Thr. 27758867 4 92 theme compositions 1107:1118 arg1 derivation 1086:1095 derivation 1086:1095 derivation of atomic compositions 1086:1118 A variety of electrospray-MS strategies using Q-TOF technology were used to define this entity, including positive and negative ion collisionally activated decomposition MS/MS, which produced unique fragmentation patterns, and high resolution accurate mass measurement to allow derivation of atomic compositions, leading to the suggestion of a taurine-containing peptidylamido-glycan structure. 26542006 6 0 theme labeled 930:936 arg1 glycans 938:944 labeled glycans 930:944 labeled glycans 930:944 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 7 1 theme screening 1126:1134 arg1 design 1136:1141 Plackett-Burman screening design 1110:1141 Plackett-Burman screening design 1110:1141 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 8 2 theme N-glycome 1443:1451 arg1 variability 1417:1427 biological variability 1406:1427 biological variability of the plasma N-glycome 1406:1451 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 6 3 theme cold 988:991 arg1 acetonitrile 997:1008 cold 96% acetonitrile 988:1008 cold 96% acetonitrile 988:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 5 4 theme silica 765:770 arg1 Bio-Gel 777:783 Bio-Gel 777:783 Bio-Gel 777:783 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 4 theme silica 765:770 arg1 filter 806:811 hydrophilic GHP filter 790:811 hydrophilic GHP filter 790:811 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 4 theme silica 765:770 arg1 phases 827:832 stationary phases 816:832 stationary phases for solid-phase extraction 816:859 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 4 theme silica 765:770 arg1 cellulose 754:762 cellulose 754:762 cellulose 754:762 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 4 theme silica 765:770 arg1 gel 772:774 silica gel 765:774 silica gel 765:774 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 4 5 theme liquid 611:616 arg1 chromatography 618:631 ultra-performance liquid chromatography 593:631 ultra-performance liquid chromatography 593:631 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 7 6 theme between-person 1222:1235 arg1 variation 1237:1245 between-day and between-person variation 1206:1245 between-day and between-person variation 1206:1245 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 4 7 theme sample 513:518 arg1 method 532:537 a high-throughput sample preparation method 495:537 a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 495:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 5 8 theme labeled 675:681 arg1 glycans 683:689 labeled glycans 675:689 labeled glycans 675:689 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 7 9 theme Plackett-Burman 1110:1124 arg1 design 1136:1141 Plackett-Burman screening design 1110:1141 Plackett-Burman screening design 1110:1141 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 4 10 theme ultra-performance 593:609 arg1 chromatography 618:631 ultra-performance liquid chromatography 593:631 ultra-performance liquid chromatography 593:631 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 3 11 theme glycans 339:345 arg1 analysis 320:327 analysis 320:327 analysis of plasma glycans 320:345 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 4 12 theme high-throughput 497:511 arg1 method 532:537 a high-throughput sample preparation method 495:537 a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 495:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 6 13 contain had 1010:1012 arg2 reproducibility 1026:1040 the highest reproducibility 1014:1040 the highest reproducibility 1014:1040 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 6 13 contain had 1010:1012 arg1 plate 962:966 GHP filter plate 951:966 GHP filter plate in combination with cold 96% acetonitrile 951:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 6 14 theme 96 993:994 arg1 % 995:995 % 995:995 % 995:995 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 7 15 theme variation 1192:1200 arg1 analysis 1169:1176 analysis 1169:1176 analysis of major step variation and between-day and between-person variation 1169:1245 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 3 16 theme wide 222:225 arg1 range 227:231 A wide range 220:231 A wide range of analytical methods 220:253 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 3 16 theme wide 222:225 arg1 available 258:266 available 258:266 available 258:266 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 4 17 theme chromatography 618:631 arg1 analysis 633:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis 543:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 543:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 7 18 theme step 1187:1190 arg1 variation 1192:1200 major step variation 1181:1200 major step variation 1181:1200 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 8 19 theme developed 1252:1260 arg1 method 1262:1267 The developed method 1248:1267 The developed method 1248:1267 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 8 19 theme developed 1252:1260 arg1 fast 1272:1275 fast 1272:1275 fast 1272:1275 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 3 20 dep IgG 310:312 arg1 mainly 303:308 mainly 303:308 mainly 303:308 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 5 21 theme stationary 816:825 arg1 Bio-Gel 777:783 Bio-Gel 777:783 Bio-Gel 777:783 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 21 theme stationary 816:825 arg1 filter 806:811 hydrophilic GHP filter 790:811 hydrophilic GHP filter 790:811 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 21 theme stationary 816:825 arg1 phases 827:832 stationary phases 816:832 stationary phases for solid-phase extraction 816:859 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 21 theme stationary 816:825 arg1 cellulose 754:762 cellulose 754:762 cellulose 754:762 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 21 theme stationary 816:825 arg1 gel 772:774 silica gel 765:774 silica gel 765:774 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 2 22 theme Aberrant 140:147 arg1 glycosylation 149:161 Aberrant glycosylation 140:161 Aberrant glycosylation 140:161 Aberrant glycosylation has been shown to be associated with different diseases. 26542006 3 23 theme analytical 236:245 arg1 methods 247:253 analytical methods 236:253 analytical methods 236:253 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 3 24 theme higher 413:418 arg1 complexity 420:429 higher complexity 413:429 higher complexity of the sample 413:443 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 0 25 theme High-throughput 0:14 arg1 glycomics 16:24 High-throughput glycomics 0:24 High-throughput glycomics: optimization of sample preparation. 0:61 High-throughput glycomics: optimization of sample preparation. 26542006 4 26 theme liquid 567:572 arg1 chromatography 574:587 hydrophilic interaction liquid chromatography 543:587 hydrophilic interaction liquid chromatography 543:587 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 3 27 theme methods 247:253 arg1 range 227:231 A wide range 220:231 A wide range of analytical methods 220:253 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 3 27 theme methods 247:253 arg1 available 258:266 available 258:266 available 258:266 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 8 28 theme long 1359:1362 arg1 period 1364:1369 long period 1359:1369 long period of time 1359:1377 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 4 29 theme interaction 555:565 arg1 chromatography 574:587 hydrophilic interaction liquid chromatography 543:587 hydrophilic interaction liquid chromatography 543:587 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 5 30 theme variation 729:737 arg1 source 719:724 the largest source 707:724 the largest source of variation 707:737 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 30 theme variation 729:737 arg1 Clean-up 663:670 Clean-up 663:670 Clean-up of labeled glycans 663:689 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 8 31 theme time 1374:1377 arg1 period 1364:1369 long period 1359:1369 long period of time 1359:1377 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 5 32 theme solid-phase 838:848 arg1 extraction 850:859 solid-phase extraction 838:859 solid-phase extraction 838:859 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 7 33 theme analysis 1169:1176 arg1 terms 1160:1164 terms 1160:1164 terms of analysis of major step variation and between-day and between-person variation 1160:1245 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 6 34 from plate 962:966 arg1 combination 971:981 combination 971:981 combination with cold 96% acetonitrile 971:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 8 35 theme plasma 1436:1441 arg1 N-glycome 1443:1451 the plasma N-glycome 1432:1451 the plasma N-glycome 1432:1451 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 0 36 theme sample 43:48 arg1 preparation 50:60 sample preparation 43:60 sample preparation 43:60 High-throughput glycomics: optimization of sample preparation. 26542006 4 37 theme chromatography 574:587 arg1 analysis 633:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis 543:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 543:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 7 38 theme major 1181:1185 arg1 variation 1192:1200 major step variation 1181:1200 major step variation 1181:1200 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 3 39 theme sample 438:443 arg1 complexity 420:429 higher complexity 413:429 higher complexity of the sample 413:443 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 5 40 theme hydrophilic 790:800 arg1 Bio-Gel 777:783 Bio-Gel 777:783 Bio-Gel 777:783 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 40 theme hydrophilic 790:800 arg1 filter 806:811 hydrophilic GHP filter 790:811 hydrophilic GHP filter 790:811 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 40 theme hydrophilic 790:800 arg1 phases 827:832 stationary phases 816:832 stationary phases for solid-phase extraction 816:859 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 40 theme hydrophilic 790:800 arg1 cellulose 754:762 cellulose 754:762 cellulose 754:762 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 40 theme hydrophilic 790:800 arg1 gel 772:774 silica gel 765:774 silica gel 765:774 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 4 41 theme method 532:537 arg1 optimization 479:490 optimization 479:490 optimization 479:490 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 4 41 theme method 532:537 arg1 development 463:473 development 463:473 development 463:473 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 3 42 theme glycan 272:277 arg1 analysis 279:286 glycan analysis 272:286 glycan analysis of antibodies (mainly IgG) 272:313 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 6 43 theme % 995:995 arg1 acetonitrile 997:1008 cold 96% acetonitrile 988:1008 cold 96% acetonitrile 988:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 3 44 theme plasma 332:337 arg1 glycans 339:345 plasma glycans 332:345 plasma glycans 332:345 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 3 45 theme additional 374:383 arg1 challenges 385:394 additional challenges 374:394 additional challenges encountered with higher complexity of the sample 374:443 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 3 46 dep antibodies 291:300 arg1 IgG 310:312 IgG 310:312 IgG 310:312 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 6 47 theme stationary 866:875 arg1 phases 877:882 All stationary phases 862:882 All stationary phases 862:882 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 6 48 theme filter 955:960 arg1 plate 962:966 GHP filter plate 951:966 GHP filter plate in combination with cold 96% acetonitrile 951:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 0 49 theme preparation 50:60 arg1 optimization 27:38 optimization 27:38 High-throughput glycomics: optimization of sample preparation. 0:61 High-throughput glycomics: optimization of sample preparation. 26542006 4 50 theme hydrophilic 543:553 arg1 chromatography 574:587 hydrophilic interaction liquid chromatography 543:587 hydrophilic interaction liquid chromatography 543:587 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 5 51 theme largest 711:717 arg1 source 719:724 the largest source 707:724 the largest source of variation 707:737 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 51 theme largest 711:717 arg1 Clean-up 663:670 Clean-up 663:670 Clean-up of labeled glycans 663:689 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 7 52 theme between-day 1206:1216 arg1 variation 1237:1245 between-day and between-person variation 1206:1245 between-day and between-person variation 1206:1245 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 1 53 theme numerous 121:128 arg1 proteins 130:137 numerous proteins 121:137 numerous proteins 121:137 Glycosylation affects structure, folding, and function of numerous proteins. 26542006 8 54 theme variability 1417:1427 arg1 detection 1393:1401 the detection 1389:1401 the detection of biological variability of the plasma N-glycome 1389:1451 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 3 55 theme antibodies 291:300 arg1 analysis 279:286 glycan analysis 272:286 glycan analysis of antibodies (mainly IgG) 272:313 A wide range of analytical methods is available for glycan analysis of antibodies (mainly IgG), but analysis of plasma glycans is less established due to additional challenges encountered with higher complexity of the sample. 26542006 8 56 theme biological 1406:1415 arg1 variability 1417:1427 biological variability 1406:1427 biological variability of the plasma N-glycome 1406:1451 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 1 57 theme proteins 130:137 arg1 structure 85:93 structure 85:93 structure 85:93 Glycosylation affects structure, folding, and function of numerous proteins. 26542006 1 57 theme proteins 130:137 arg1 folding 96:102 folding 96:102 folding 96:102 Glycosylation affects structure, folding, and function of numerous proteins. 26542006 1 57 theme proteins 130:137 arg1 function 109:116 function 109:116 function 109:116 Glycosylation affects structure, folding, and function of numerous proteins. 26542006 4 58 theme plasma 645:650 arg1 N-glycans 652:660 plasma N-glycans 645:660 plasma N-glycans 645:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 6 59 with combination 971:981 arg1 acetonitrile 997:1008 cold 96% acetonitrile 988:1008 cold 96% acetonitrile 988:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 2 60 theme different 200:208 arg1 diseases 210:217 different diseases 200:217 different diseases 200:217 Aberrant glycosylation has been shown to be associated with different diseases. 26542006 8 61 theme good 1331:1334 arg1 reproducibility 1336:1350 very good reproducibility 1326:1350 very good reproducibility 1326:1350 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 8 62 contain has 1322:1324 arg1 it 1319:1320 it 1319:1320 it 1319:1320 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 8 62 contain has 1322:1324 arg2 reproducibility 1336:1350 very good reproducibility 1326:1350 very good reproducibility 1326:1350 The developed method is fast, cost-effective, and easy to perform, and it has very good reproducibility during long period of time, enabling the detection of biological variability of the plasma N-glycome. 26542006 6 63 theme highest 1018:1024 arg1 reproducibility 1026:1040 the highest reproducibility 1014:1040 the highest reproducibility 1014:1040 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 7 64 theme variation 1237:1245 arg1 analysis 1169:1176 analysis 1169:1176 analysis of major step variation and between-day and between-person variation 1169:1245 The method was further optimized with Plackett-Burman screening design and validated in terms of analysis of major step variation and between-day and between-person variation. 26542006 0 65 dep glycomics 16:24 arg1 optimization 27:38 optimization 27:38 High-throughput glycomics: optimization of sample preparation. 0:61 High-throughput glycomics: optimization of sample preparation. 26542006 5 66 theme GHP 802:804 arg1 Bio-Gel 777:783 Bio-Gel 777:783 Bio-Gel 777:783 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 66 theme GHP 802:804 arg1 filter 806:811 hydrophilic GHP filter 790:811 hydrophilic GHP filter 790:811 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 66 theme GHP 802:804 arg1 phases 827:832 stationary phases 816:832 stationary phases for solid-phase extraction 816:859 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 66 theme GHP 802:804 arg1 cellulose 754:762 cellulose 754:762 cellulose 754:762 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 66 theme GHP 802:804 arg1 gel 772:774 silica gel 765:774 silica gel 765:774 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 6 67 theme glycans 938:944 arg1 purification 914:925 purification 914:925 purification of labeled glycans 914:944 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 4 68 theme N-glycans 652:660 arg1 analysis 633:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis 543:640 hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 543:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26542006 5 69 theme glycans 683:689 arg1 source 719:724 the largest source 707:724 the largest source of variation 707:737 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 5 69 theme glycans 683:689 arg1 Clean-up 663:670 Clean-up 663:670 Clean-up of labeled glycans 663:689 Clean-up of labeled glycans was found to be the largest source of variation, and we tested cellulose, silica gel, Bio-Gel, and hydrophilic GHP filter as stationary phases for solid-phase extraction. 26542006 6 70 theme GHP 951:953 arg1 plate 962:966 GHP filter plate 951:966 GHP filter plate in combination with cold 96% acetonitrile 951:1008 All stationary phases were shown to be suitable for purification of labeled glycans, but GHP filter plate in combination with cold 96% acetonitrile had the highest reproducibility and was easiest to work with. 26542006 4 71 theme preparation 520:530 arg1 method 532:537 a high-throughput sample preparation method 495:537 a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans 495:660 Here we describe development and optimization of a high-throughput sample preparation method for hydrophilic interaction liquid chromatography and ultra-performance liquid chromatography analysis of plasma N-glycans. 26538210 0 0 theme Structures 100:109 arg1 Multiplicity 66:77 a Multiplicity 64:77 a Multiplicity of Isomeric N-Glycan Structures 64:109 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 8 1 theme complex 1451:1457 arg1 glycomes 1472:1479 complex invertebrate glycomes 1451:1479 complex invertebrate glycomes 1451:1479 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 2 2 theme fucose 387:392 arg1 residues 394:401 Up to four fucose residues 376:401 Up to four fucose residues 376:401 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 2 2 theme fucose 387:392 arg1 fucose 434:439 fucose 434:439 fucose 434:439 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 5 3 from α1,6-fucose 925:935 arg1 galactose 988:996 the bisecting galactose 974:996 the bisecting galactose 974:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 7 4 dep 13 1165:1166 arg1 to 1162:1163 to 1162:1163 to 1162:1163 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 0 5 theme N-Glycan 91:98 arg1 Structures 100:109 Isomeric N-Glycan Structures 82:109 Isomeric N-Glycan Structures 82:109 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 8 6 theme multiple 1385:1392 arg1 approaches 1394:1403 multiple approaches 1385:1403 multiple approaches 1385:1403 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 4 7 theme released 665:672 arg1 N-glycans 674:682 Enzymatically released N-glycans 651:682 Enzymatically released N-glycans 651:682 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 7 8 theme different 1106:1114 arg1 locations 1116:1124 The different locations 1102:1124 The different locations 1102:1124 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 3 9 from complexity 494:503 arg1 type 517:520 the wild type 508:520 the wild type 508:520 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 10 theme mutant 544:549 arg1 strains 551:557 three mutant strains 538:557 three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8) 538:648 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 6 11 theme fut-8 1087:1091 arg1 strains 1093:1099 fut-8 strains 1087:1099 fut-8 strains 1087:1099 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 4 12 from combination 729:739 arg1 subject 689:695 subject 689:695 subject 689:695 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 2 13 dep four 382:385 arg1 to 379:380 to 379:380 to 379:380 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 3 14 theme core 576:579 arg1 fucosyltransferases 581:599 two core fucosyltransferases 572:599 two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8) 572:648 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 4 15 theme various 746:752 arg1 treatments 754:763 various treatments 746:763 various treatments 746:763 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 5 16 theme mutant 829:834 arg1 complex 849:855 the most complex 840:855 the most complex of the three double-mutant strains 840:890 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 16 theme mutant 829:834 arg1 N-glycome 800:808 The N-glycome 796:808 The N-glycome of the fut-1;fut-6 mutant 796:834 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 3 17 dep fucosyltransferases 581:599 arg1 fut-1 607:611 fut-1 607:611 fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8 607:647 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 17 dep fucosyltransferases 581:599 arg1 each 601:604 each 601:604 each 601:604 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 8 18 theme individual 1415:1424 arg1 structures 1433:1442 individual glycan structures 1415:1442 individual glycan structures 1415:1442 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 5 19 theme α1,6-fucose 925:935 arg1 extension 903:911 the extension 899:911 the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 899:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 20 theme fut-6 823:827 arg1 mutant 829:834 the fut-1;fut-6 mutant 813:834 the fut-1;fut-6 mutant 813:834 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 2 21 theme C. 471:472 arg1 elegans 474:480 C. elegans 471:480 C. elegans 471:480 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 1 22 from degree 151:156 arg1 N-glycome 179:187 the N-glycome 175:187 the N-glycome of the model nematode Caenorhabditis elegans 175:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 4 23 dep HPLC 700:703 arg1 MS/MS 719:723 MS/MS 719:723 MS/MS 719:723 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 7 24 theme isomeric 1168:1175 arg1 many 1189:1192 many 1189:1192 many 1189:1192 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 7 24 theme isomeric 1168:1175 arg1 structures 1177:1186 up to 13 isomeric structures 1159:1186 up to 13 isomeric structures 1159:1186 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 2 25 theme different 412:420 arg1 linkages 422:429 five different linkages 407:429 five different linkages of fucose 407:439 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 1 26 theme altered 300:306 arg1 profiles 317:324 radically altered N-glycan profiles 290:324 radically altered N-glycan profiles 290:324 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 1 27 theme Recent 112:117 arg1 studies 119:125 Recent studies 112:125 Recent studies 112:125 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 3 28 from fucosyltransferases 581:599 arg1 defective 559:567 defective 559:567 defective 559:567 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 1 29 theme model 196:200 arg1 elegans 226:232 the model nematode Caenorhabditis elegans 192:232 the model nematode Caenorhabditis elegans 192:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 0 30 theme Fucosyltransferase 30:47 arg1 Mutants 49:55 Caenorhabditis Fucosyltransferase Mutants 15:55 Caenorhabditis Fucosyltransferase Mutants 15:55 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 1 31 theme N-glycan 308:315 arg1 profiles 317:324 radically altered N-glycan profiles 290:324 radically altered N-glycan profiles 290:324 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 8 32 theme simple 1336:1341 arg1 " 1342:1342 a "simple" 1333:1342 a "simple" nematode 1333:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 5 33 from fucose 964:969 arg1 galactose 988:996 the bisecting galactose 974:996 the bisecting galactose 974:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 3 34 theme defective 559:567 arg1 strains 551:557 three mutant strains 538:557 three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8) 538:648 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 8 35 theme high 1286:1289 arg1 variability 1291:1301 the high variability 1282:1301 the high variability of the N-glycomic capacity of a "simple" nematode 1282:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 1 36 theme nematode 202:209 arg1 elegans 226:232 the model nematode Caenorhabditis elegans 192:232 the model nematode Caenorhabditis elegans 192:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 0 37 theme Caenorhabditis 15:28 arg1 Mutants 49:55 Caenorhabditis Fucosyltransferase Mutants 15:55 Caenorhabditis Fucosyltransferase Mutants 15:55 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 4 38 with combination 729:739 arg1 treatments 754:763 various treatments 746:763 various treatments 746:763 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 5 39 theme core 920:923 arg1 α1,6-fucose 925:935 the core α1,6-fucose 916:935 the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 916:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 6 40 theme fut-1 1065:1069 arg1 N-glycans 1048:1056 N-glycans 1048:1056 N-glycans of the fut-1 1048:1069 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 5 41 from presence 952:959 arg1 galactose 988:996 the bisecting galactose 974:996 the bisecting galactose 974:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 1 42 theme Caenorhabditis 211:224 arg1 elegans 226:232 the model nematode Caenorhabditis elegans 192:232 the model nematode Caenorhabditis elegans 192:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 3 43 theme wild 512:515 arg1 type 517:520 the wild type 508:520 the wild type 508:520 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 6 44 located found 1039:1043 arg1 N-glycans 1048:1056 N-glycans 1048:1056 N-glycans of the fut-1 1048:1069 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 6 44 located found 1039:1043 arg2 fut-8 1071:1075 fut-8 1071:1075 fut-8 1071:1075 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 6 44 located found 1039:1043 arg2 fut-6 1081:1085 fut-6 1081:1085 fut-6 1081:1085 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 6 44 located found 1039:1043 arg2 strains 1093:1099 fut-8 strains 1087:1099 fut-8 strains 1087:1099 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 6 44 located found 1039:1043 arg1 contrast 1002:1009 contrast 1002:1009 contrast 1002:1009 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 6 44 located found 1039:1043 arg2 fucoses 1026:1032 maximally two fucoses 1012:1032 maximally two fucoses 1012:1032 In contrast, maximally two fucoses were found on N-glycans of the fut-1;fut-8 and fut-6;fut-8 strains. 26538210 5 45 theme presence 952:959 arg1 extension 903:911 the extension 899:911 the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 899:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 3 46 dep fut-1 607:611 arg1 fut-6 613:617 fut-6 613:617 fut-6 613:617 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 46 dep fut-1 607:611 arg1 fut-8 643:647 fut-8 643:647 fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8 607:647 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 46 dep fut-1 607:611 arg1 fut-8 626:630 fut-8 626:630 fut-8 626:630 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 46 dep fut-1 607:611 arg1 fut-6 637:641 fut-6 637:641 fut-6 637:641 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 3 46 dep fut-1 607:611 arg1 fut-1 620:624 fut-1 620:624 fut-1 620:624 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 1 47 theme elegans 226:232 arg1 N-glycome 179:187 the N-glycome 175:187 the N-glycome of the model nematode Caenorhabditis elegans 175:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 0 48 theme Mutants 49:55 arg1 Comparisons 0:10 Comparisons 0:10 Comparisons of Caenorhabditis Fucosyltransferase Mutants 0:55 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 8 49 theme invertebrate 1459:1470 arg1 glycomes 1472:1479 complex invertebrate glycomes 1451:1479 complex invertebrate glycomes 1451:1479 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 8 50 theme " 1342:1342 arg1 nematode 1344:1351 a "simple" nematode 1333:1351 a "simple" nematode 1333:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 4 51 theme structural 776:785 arg1 details 787:793 structural details 776:793 structural details 776:793 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 2 52 theme fucose 434:439 arg1 linkages 422:429 five different linkages 407:429 five different linkages of fucose 407:439 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 2 52 theme fucose 434:439 arg1 residues 394:401 Up to four fucose residues 376:401 Up to four fucose residues 376:401 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 2 52 theme fucose 434:439 arg1 fucose 434:439 fucose 434:439 fucose 434:439 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 1 53 theme minor 339:343 arg1 effects 367:373 only minor biological phenotypic effects 334:373 only minor biological phenotypic effects 334:373 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 8 54 theme glycan 1426:1431 arg1 structures 1433:1442 individual glycan structures 1415:1442 individual glycan structures 1415:1442 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 5 55 theme fucose 964:969 arg1 presence 952:959 the presence 948:959 the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 916:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 55 theme fucose 964:969 arg1 α1,6-fucose 925:935 the core α1,6-fucose 916:935 the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 916:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 2 56 theme elegans 474:480 arg1 N-glycans 458:466 the N-glycans 454:466 the N-glycans of C. elegans 454:480 Up to four fucose residues and five different linkages of fucose are known on the N-glycans of C. elegans. 26538210 1 57 theme biological 345:354 arg1 effects 367:373 only minor biological phenotypic effects 334:373 only minor biological phenotypic effects 334:373 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 8 58 theme nematode 1344:1351 arg1 capacity 1321:1328 the N-glycomic capacity 1306:1328 the N-glycomic capacity of a "simple" nematode 1306:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 3 59 from defective 559:567 arg1 fucosyltransferases 581:599 two core fucosyltransferases 572:599 two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8) 572:648 Due to the complexity in the wild type, we established three mutant strains defective in two core fucosyltransferases each (fut-1;fut-6, fut-1;fut-8, and fut-6;fut-8). 26538210 1 60 theme phenotypic 356:365 arg1 effects 367:373 only minor biological phenotypic effects 334:373 only minor biological phenotypic effects 334:373 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 5 61 theme bisecting 978:986 arg1 galactose 988:996 the bisecting galactose 974:996 the bisecting galactose 974:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 8 62 theme N-glycomic 1310:1319 arg1 capacity 1321:1328 the N-glycomic capacity 1306:1328 the N-glycomic capacity of a "simple" nematode 1306:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 1 63 theme remarkable 140:149 arg1 degree 151:156 a remarkable degree 138:156 a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans 138:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 5 64 from galactose 988:996 arg1 presence 952:959 the presence 948:959 the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 916:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 64 from galactose 988:996 arg1 α1,6-fucose 925:935 the core α1,6-fucose 916:935 the core α1,6-fucose as well as the presence of fucose on the bisecting galactose 916:996 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 65 theme strains 884:890 arg1 complex 849:855 the most complex 840:855 the most complex of the three double-mutant strains 840:890 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 65 theme strains 884:890 arg1 N-glycome 800:808 The N-glycome 796:808 The N-glycome of the fut-1;fut-6 mutant 796:834 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 4 66 from subject 689:695 arg1 combination 729:739 combination 729:739 combination with various treatments 729:763 Enzymatically released N-glycans were subject to HPLC and MALDI-TOF MS/MS, in combination with various treatments, to verify structural details. 26538210 5 67 theme double-mutant 870:882 arg1 strains 884:890 the three double-mutant strains 860:890 the three double-mutant strains 860:890 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 0 68 theme Isomeric 82:89 arg1 Structures 100:109 Isomeric N-Glycan Structures 82:109 Isomeric N-Glycan Structures 82:109 Comparisons of Caenorhabditis Fucosyltransferase Mutants Reveal a Multiplicity of Isomeric N-Glycan Structures. 26538210 5 69 theme fut-1 817:821 arg1 mutant 829:834 the fut-1;fut-6 mutant 813:834 the fut-1;fut-6 mutant 813:834 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 7 70 theme single 1242:1247 arg1 masses 1249:1254 some single masses 1237:1254 some single masses 1237:1254 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 5 71 theme most 844:847 arg1 complex 849:855 the most complex 840:855 the most complex of the three double-mutant strains 840:890 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 5 71 theme most 844:847 arg1 N-glycome 800:808 The N-glycome 796:808 The N-glycome of the fut-1;fut-6 mutant 796:834 The N-glycome of the fut-1;fut-6 mutant was the most complex of the three double-mutant strains due to the extension of the core α1,6-fucose as well as the presence of fucose on the bisecting galactose. 26538210 1 72 theme glycosylation-relevant 247:268 arg1 genes 270:274 glycosylation-relevant genes 247:274 glycosylation-relevant genes 247:274 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 7 73 theme fucose 1141:1146 arg1 capping 1130:1136 capping 1130:1136 capping of fucose 1130:1146 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 7 73 theme fucose 1141:1146 arg1 locations 1116:1124 The different locations 1102:1124 The different locations 1102:1124 The different locations and capping of fucose meant that up to 13 isomeric structures, many highly galactosylated, were determined for some single masses. 26538210 8 74 theme capacity 1321:1328 arg1 variability 1291:1301 the high variability 1282:1301 the high variability of the N-glycomic capacity of a "simple" nematode 1282:1351 These data not only show the high variability of the N-glycomic capacity of a "simple" nematode but also exemplify the need for multiple approaches to reveal individual glycan structures within complex invertebrate glycomes. 26538210 1 75 theme plasticity 161:170 arg1 degree 151:156 a remarkable degree 138:156 a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans 138:232 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26538210 1 76 theme genes 270:274 arg1 ablation 235:242 ablation 235:242 ablation of glycosylation-relevant genes 235:274 Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. 26537504 6 0 theme variants 1020:1027 arg1 application 967:977 The application 963:977 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs 963:1074 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 7 1 theme vascular 1381:1388 arg1 lesions 1390:1396 the hemorrhagic vascular lesions 1365:1396 the hemorrhagic vascular lesions observed 1365:1405 On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. 26537504 6 2 theme pro-inflammatory 1092:1107 arg1 reactions 1127:1135 the pro-inflammatory and immunological reactions 1088:1135 the pro-inflammatory and immunological reactions 1088:1135 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 2 theme pro-inflammatory 1092:1107 arg1 due 1209:1211 due 1209:1211 due 1209:1211 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 1 3 theme females 245:251 arg1 abdomen 228:234 the abdomen 224:234 the abdomen of adult females of the neo-tropical moth 224:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 3 4 theme N-glycosylation 664:678 arg1 site 680:683 an N-glycosylation site 661:683 an N-glycosylation site 661:683 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 2 5 contain contain 448:454 arg1 setae 442:446 The setae 438:446 The setae 438:446 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 2 5 contain contain 448:454 arg2 protease 488:495 a pro-inflammatory glycosylated protease 456:495 a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects 456:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 0 6 theme Hylesia 125:131 arg1 Cramer 142:147 Cramer [1775 142:153 Cramer [1775 142:153 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 6 theme Hylesia 125:131 arg1 metabus 133:139 the neotropical moth Hylesia metabus 104:139 the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 104:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 6 7 theme groups 1247:1252 arg1 presence 1220:1227 the presence 1216:1227 the presence of sulfate-linked groups and not on disulfide bonds 1216:1279 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 1 8 theme generalized 369:379 arg1 malaise 381:387 generalized malaise 369:387 generalized malaise 369:387 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 0 9 dep Lepidoptera 158:168 arg1 Saturniidae 171:181 Saturniidae 171:181 Saturniidae 171:181 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 6 10 theme disseminated 1143:1154 arg1 fibrin 1156:1161 disseminated fibrin 1143:1161 disseminated fibrin deposition 1143:1172 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 5 11 theme core 957:960 arg1 residue 942:948 the α(1-6)-mannose residue 923:948 the α(1-6)-mannose residue of the core 923:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 5 11 theme core 957:960 arg1 core 957:960 the core 953:960 the core 953:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 1 12 theme ocular 406:411 arg1 lepidopterism 422:434 lepidopterism 422:434 lepidopterism 422:434 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 1 12 theme ocular 406:411 arg1 lesions 413:419 occasionally ocular lesions 393:419 occasionally ocular lesions (lepidopterism) 393:435 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 2 13 theme homologous 497:506 arg1 protease 488:495 a pro-inflammatory glycosylated protease 456:495 a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects 456:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 6 14 theme immunological 1113:1125 arg1 reactions 1127:1135 the pro-inflammatory and immunological reactions 1088:1135 the pro-inflammatory and immunological reactions 1088:1135 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 14 theme immunological 1113:1125 arg1 due 1209:1211 due 1209:1211 due 1209:1211 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 15 theme toxin 1339:1343 arg1 S-alkylation 1319:1330 S-alkylation 1319:1330 S-alkylation 1319:1330 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 15 theme toxin 1339:1343 arg1 reduction 1305:1313 reduction 1305:1313 reduction 1305:1313 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 2 16 theme glycosylated 475:486 arg1 protease 488:495 a pro-inflammatory glycosylated protease 456:495 a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects 456:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 6 17 dep deposition 1163:1172 arg1 e.g. 1138:1141 e.g. 1138:1141 e.g. 1138:1141 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 0 18 theme serine 83:88 arg1 protease 90:97 a serine protease 81:97 a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 81:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 1 19 theme urticating 311:320 arg1 dermatitis 322:331 an urticating dermatitis 308:331 an urticating dermatitis 308:331 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 2 20 theme serine 521:526 arg1 proteases 528:536 other S1A serine proteases 511:536 other S1A serine proteases of insects 511:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 4 21 from GlcNAc 806:811 arg1 -fucosylated 777:788 -fucosylated 777:788 -fucosylated 777:788 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 2 22 theme other 511:515 arg1 proteases 528:536 other S1A serine proteases 511:536 other S1A serine proteases of insects 511:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 0 23 theme neotropical 108:118 arg1 Cramer 142:147 Cramer [1775 142:153 Cramer [1775 142:153 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 23 theme neotropical 108:118 arg1 metabus 133:139 the neotropical moth Hylesia metabus 104:139 the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 104:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 24 from implications 43:54 arg1 protease 90:97 a serine protease 81:97 a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 81:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 4 25 theme paucimannosidic 696:710 arg1 N-glycans 712:720 Five main paucimannosidic N-glycans 686:720 Five main paucimannosidic N-glycans 686:720 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 0 26 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization 0:26 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 6 27 from model 1055:1059 arg1 pigs 1071:1074 guinea pigs 1064:1074 guinea pigs 1064:1074 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 4 28 gly -fucosylated 777:788 arg2 GlcNAc 806:811 the proximal GlcNAc 793:811 the proximal GlcNAc 793:811 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 4 28 gly -fucosylated 777:788 arg1 which 748:752 which 748:752 which 748:752 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 4 28 gly -fucosylated 777:788 arg1 three 739:743 three 739:743 three 739:743 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 1 29 theme neo-tropical 260:271 arg1 moth 273:276 the neo-tropical moth 256:276 the neo-tropical moth 256:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 2 30 gly glycosylated 475:486 arg1 protease 488:495 a pro-inflammatory glycosylated protease 456:495 a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects 456:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 3 31 gly N-glycosylation 664:678 arg2 site 680:683 an N-glycosylation site 661:683 an N-glycosylation site 661:683 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 6 32 from presence 1220:1227 arg1 bonds 1275:1279 disulfide bonds 1265:1279 disulfide bonds 1265:1279 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 1 33 with Contact 185:191 arg1 setae 213:217 the urticating setae 198:217 the urticating setae from the abdomen of adult females of the neo-tropical moth 198:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 7 34 theme proteolytic 1429:1439 arg1 activity 1441:1448 the proteolytic activity 1425:1448 the proteolytic activity of the toxin 1425:1461 On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. 26537504 5 35 theme residue 942:948 arg1 6-position 909:918 the 6-position 905:918 the 6-position of the α(1-6)-mannose residue of the core 905:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 8 36 theme N-glycan 1470:1477 arg1 sulfation 1479:1487 N-glycan sulfation 1470:1487 N-glycan sulfation 1470:1487 Thus, N-glycan sulfation may constitute a defense mechanism against predators. 26537504 3 37 theme % 624:624 arg1 O 633:633 40% H2 (18)O 622:633 40% H2 (18)O 622:633 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 0 38 theme sulfated 59:66 arg1 N-glycans 68:76 sulfated N-glycans 59:76 sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 59:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 3 39 theme buffer 601:606 arg1 presence 587:594 the presence 583:594 the presence of a buffer prepared with 40% H2 (18)O 583:633 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 1 40 theme urticating 202:211 arg1 setae 213:217 the urticating setae 198:217 the urticating setae from the abdomen of adult females of the neo-tropical moth 198:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 6 41 theme toxin 1036:1040 arg1 variants 1020:1027 chemically and enzymatically modified variants 982:1027 chemically and enzymatically modified variants of the toxin 982:1040 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 7 42 theme hemorrhagic 1369:1379 arg1 lesions 1390:1396 the hemorrhagic vascular lesions 1365:1396 the hemorrhagic vascular lesions observed 1365:1405 On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. 26537504 6 43 theme modified 1011:1018 arg1 variants 1020:1027 chemically and enzymatically modified variants 982:1027 chemically and enzymatically modified variants of the toxin 982:1040 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 44 link sulfate-linked 1232:1245 arg1 groups 1247:1252 sulfate-linked groups 1232:1252 sulfate-linked groups 1232:1252 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 1 45 theme adult 239:243 arg1 females 245:251 adult females 239:251 adult females 239:251 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 5 46 theme N-glycans 846:854 arg1 species 868:874 anionic species 860:874 anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core 860:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 5 46 theme N-glycans 846:854 arg1 portion 829:835 A considerable portion 814:835 A considerable portion of these N-glycans 814:854 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 6 47 theme neutrophils 1192:1202 arg1 activation 1178:1187 activation 1178:1187 activation of neutrophils 1178:1202 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 47 theme neutrophils 1192:1202 arg1 deposition 1163:1172 disseminated fibrin deposition 1143:1172 disseminated fibrin deposition 1143:1172 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 7 48 theme other 1353:1357 arg1 hand 1359:1362 the other hand 1349:1362 the other hand 1349:1362 On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. 26537504 3 49 theme site 680:683 arg1 assignment 647:656 the assignment 643:656 the assignment of an N-glycosylation site 643:683 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 3 50 theme PNGase 571:576 arg1 F 578:578 PNGase F 571:578 PNGase F 571:578 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 5 51 theme considerable 816:827 arg1 species 868:874 anionic species 860:874 anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core 860:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 5 51 theme considerable 816:827 arg1 portion 829:835 A considerable portion 814:835 A considerable portion of these N-glycans 814:854 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 4 52 theme proximal 797:804 arg1 GlcNAc 806:811 the proximal GlcNAc 793:811 the proximal GlcNAc 793:811 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 6 53 theme guinea 1064:1069 arg1 pigs 1071:1074 guinea pigs 1064:1074 guinea pigs 1064:1074 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 54 from application 967:977 arg1 model 1055:1059 an animal model 1045:1059 an animal model in guinea pigs 1045:1074 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 2 55 theme insects 541:547 arg1 proteases 528:536 other S1A serine proteases 511:536 other S1A serine proteases of insects 511:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 0 56 from protease 90:97 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization 0:26 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 56 from protease 90:97 arg1 implications 43:54 biological implications 32:54 biological implications 32:54 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 1 57 from abdomen 228:234 arg1 setae 213:217 the urticating setae 198:217 the urticating setae from the abdomen of adult females of the neo-tropical moth 198:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 6 58 theme fibrin 1156:1161 arg1 deposition 1163:1172 disseminated fibrin deposition 1143:1172 disseminated fibrin deposition 1143:1172 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 59 theme sulfate-linked 1232:1245 arg1 groups 1247:1252 sulfate-linked groups 1232:1252 sulfate-linked groups 1232:1252 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 5 60 theme anionic 860:866 arg1 species 868:874 anionic species 860:874 anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core 860:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 5 60 theme anionic 860:866 arg1 portion 829:835 A considerable portion 814:835 A considerable portion of these N-glycans 814:854 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 3 61 from Deglycosylation 550:564 arg1 presence 587:594 the presence 583:594 the presence of a buffer prepared with 40% H2 (18)O 583:633 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 3 62 gly Deglycosylation 550:564 arg1 presence 587:594 the presence 583:594 the presence of a buffer prepared with 40% H2 (18)O 583:633 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 0 63 dep metabus 133:139 arg1 Lepidoptera 158:168 Lepidoptera 158:168 Lepidoptera 158:168 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 2 64 theme pro-inflammatory 458:473 arg1 protease 488:495 a pro-inflammatory glycosylated protease 456:495 a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects 456:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 0 65 from characterization 11:26 arg1 protease 90:97 a serine protease 81:97 a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 81:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 7 66 theme toxin 1457:1461 arg1 activity 1441:1448 the proteolytic activity 1425:1448 the proteolytic activity of the toxin 1425:1461 On the other hand, the hemorrhagic vascular lesions observed are attributed to the proteolytic activity of the toxin. 26537504 2 67 theme S1A 517:519 arg1 proteases 528:536 other S1A serine proteases 511:536 other S1A serine proteases of insects 511:547 The setae contain a pro-inflammatory glycosylated protease homologous to other S1A serine proteases of insects. 26537504 0 68 from N-glycans 68:76 arg1 protease 90:97 a serine protease 81:97 a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 81:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 68 from N-glycans 68:76 arg1 metabus 133:139 the neotropical moth Hylesia metabus 104:139 the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 104:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 68 from N-glycans 68:76 arg1 Cramer 142:147 Cramer [1775 142:153 Cramer [1775 142:153 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 69 theme moth 120:123 arg1 Cramer 142:147 Cramer [1775 142:153 Cramer [1775 142:153 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 69 theme moth 120:123 arg1 metabus 133:139 the neotropical moth Hylesia metabus 104:139 the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 104:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 3 70 with Deglycosylation 550:564 arg1 F 578:578 PNGase F 571:578 PNGase F 571:578 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 1 71 theme intense 351:357 arg1 pruritus 359:366 intense pruritus 351:366 intense pruritus 351:366 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 6 72 dep reduction 1305:1313 arg1 the 1301:1303 the 1301:1303 the 1301:1303 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 4 73 theme main 691:694 arg1 N-glycans 712:720 Five main paucimannosidic N-glycans 686:720 Five main paucimannosidic N-glycans 686:720 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 0 74 theme biological 32:41 arg1 implications 43:54 biological implications 32:54 biological implications 32:54 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 3 75 theme H2 626:627 arg1 O 633:633 40% H2 (18)O 622:633 40% H2 (18)O 622:633 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 1 76 theme moth 273:276 arg1 abdomen 228:234 the abdomen 224:234 the abdomen of adult females of the neo-tropical moth 224:276 Contact with the urticating setae from the abdomen of adult females of the neo-tropical moth Hylesia metabus gives rise to an urticating dermatitis, characterized by intense pruritus, generalized malaise and occasionally ocular lesions (lepidopterism). 26537504 5 77 theme -mannose 933:940 arg1 residue 942:948 the α(1-6)-mannose residue 923:948 the α(1-6)-mannose residue of the core 923:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 5 77 theme -mannose 933:940 arg1 core 957:960 the core 953:960 the core 953:960 A considerable portion of these N-glycans are anionic species sulfated on either the 4- or the 6-position of the α(1-6)-mannose residue of the core. 26537504 3 78 theme 40 622:623 arg1 % 624:624 % 624:624 % 624:624 Deglycosylation with PNGase F in the presence of a buffer prepared with 40% H2 (18)O allowed the assignment of an N-glycosylation site. 26537504 6 79 theme disulfide 1265:1273 arg1 bonds 1275:1279 disulfide bonds 1265:1279 disulfide bonds 1265:1279 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 0 80 theme N-glycans 68:76 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization 0:26 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 80 theme N-glycans 68:76 arg1 implications 43:54 biological implications 32:54 biological implications 32:54 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 6 81 theme animal 1048:1053 arg1 model 1055:1059 an animal model 1045:1059 an animal model in guinea pigs 1045:1074 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 8 82 theme defense 1506:1512 arg1 mechanism 1514:1522 a defense mechanism 1504:1522 a defense mechanism against predators 1504:1540 Thus, N-glycan sulfation may constitute a defense mechanism against predators. 26537504 4 83 theme which 748:752 arg1 which 748:752 which 748:752 which 748:752 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 4 83 theme which 748:752 arg1 three 739:743 three 739:743 three 739:743 Five main paucimannosidic N-glycans were identified, three of which were exclusively α(1-6)-fucosylated at the proximal GlcNAc. 26537504 6 84 attach presence 1220:1227 arg2 groups 1247:1252 sulfate-linked groups 1232:1252 sulfate-linked groups 1232:1252 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 6 84 attach presence 1220:1227 arg1 bonds 1275:1279 disulfide bonds 1265:1279 disulfide bonds 1265:1279 The application of chemically and enzymatically modified variants of the toxin in an animal model in guinea pigs showed that the pro-inflammatory and immunological reactions, e.g. disseminated fibrin deposition and activation of neutrophils, are due to the presence of sulfate-linked groups and not on disulfide bonds, as demonstrated by the reduction and S-alkylation of the toxin. 26537504 0 85 from metabus 133:139 arg1 protease 90:97 a serine protease 81:97 a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 81:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 26537504 0 85 from metabus 133:139 arg1 N-glycans 68:76 sulfated N-glycans 59:76 sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae) 59:182 Structural characterization and biological implications of sulfated N-glycans in a serine protease from the neotropical moth Hylesia metabus (Cramer [1775]) (Lepidoptera: Saturniidae). 28050839 3 0 theme Carbohydrate 301:312 arg1 Electrophoresis 318:332 Carbohydrate gel Electrophoresis 301:332 Carbohydrate gel Electrophoresis 301:332 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 3 1 dep analyze 369:375 arg1 Goubet 416:421 Goubet 416:421 Goubet 416:421 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 3 2 theme simple 342:347 arg1 technique 356:364 a simple, rapid technique 340:364 a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002) 340:459 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 1 3 theme plant 65:69 arg1 wall 76:79 The plant cell wall 61:79 The plant cell wall 61:79 The plant cell wall is composed of many complex polysaccharides. 28050839 1 4 theme cell 71:74 arg1 wall 76:79 The plant cell wall 61:79 The plant cell wall 61:79 The plant cell wall is composed of many complex polysaccharides. 28050839 3 5 theme Anal 431:434 arg1 Biochem 436:442 Anal Biochem 300:53-68 431:452 Anal Biochem 300:53-68 431:452 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 3 6 theme polysaccharide 377:390 arg1 quantity 392:399 polysaccharide quantity 377:399 polysaccharide quantity 377:399 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 0 7 theme Structural 0:9 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of Cell Wall Polysaccharides 0:47 Structural Analysis of Cell Wall Polysaccharides Using PACE. 28050839 2 8 dep composition 130:140 arg1 The 126:128 The 126:128 The 126:128 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 3 9 dep Goubet 416:421 arg1 al. 426:428 Goubet et al., Anal Biochem 300:53-68, 2002 416:458 al. 426:428 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 3 9 dep Goubet 416:421 arg1 Biochem 436:442 Anal Biochem 300:53-68 431:452 Anal Biochem 300:53-68 431:452 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 0 10 theme Cell 23:26 arg1 Polysaccharides 33:47 Cell Wall Polysaccharides 23:47 Cell Wall Polysaccharides 23:47 Structural Analysis of Cell Wall Polysaccharides Using PACE. 28050839 3 11 theme Polysaccharide 271:284 arg1 Analysis 286:293 Polysaccharide Analysis 271:293 Polysaccharide Analysis 271:293 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 0 12 theme Polysaccharides 33:47 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of Cell Wall Polysaccharides 0:47 Structural Analysis of Cell Wall Polysaccharides Using PACE. 28050839 2 13 theme polysaccharides 163:177 arg1 composition 130:140 composition 130:140 composition 130:140 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 13 theme polysaccharides 163:177 arg1 structure 146:154 structure 146:154 structure 146:154 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 14 theme cell 232:235 arg1 function 237:244 cell function 232:244 cell function 232:244 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 0 15 theme Wall 28:31 arg1 Polysaccharides 33:47 Cell Wall Polysaccharides 23:47 Cell Wall Polysaccharides 23:47 Structural Analysis of Cell Wall Polysaccharides Using PACE. 28050839 3 16 dep simple 342:347 arg1 rapid 350:354 rapid 350:354 rapid 350:354 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 1 17 theme many 96:99 arg1 polysaccharides 109:123 many complex polysaccharides 96:123 many complex polysaccharides 96:123 The plant cell wall is composed of many complex polysaccharides. 28050839 3 18 theme gel 314:316 arg1 Electrophoresis 318:332 Carbohydrate gel Electrophoresis 301:332 Carbohydrate gel Electrophoresis 301:332 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 28050839 2 19 theme cell 220:223 arg1 shape 225:229 cell shape 220:229 cell shape 220:229 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 1 20 theme complex 101:107 arg1 polysaccharides 109:123 many complex polysaccharides 96:123 many complex polysaccharides 96:123 The plant cell wall is composed of many complex polysaccharides. 28050839 2 21 theme cell 194:197 arg1 properties 199:208 various cell properties 186:208 various cell properties including cell shape, cell function and cell adhesion 186:262 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 21 theme cell 194:197 arg1 shape 225:229 cell shape 220:229 cell shape 220:229 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 21 theme cell 194:197 arg1 function 237:244 cell function 232:244 cell function 232:244 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 21 theme cell 194:197 arg1 adhesion 255:262 cell adhesion 250:262 cell adhesion 250:262 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 22 theme cell 250:253 arg1 adhesion 255:262 cell adhesion 250:262 cell adhesion 250:262 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 23 theme various 186:192 arg1 properties 199:208 various cell properties 186:208 various cell properties including cell shape, cell function and cell adhesion 186:262 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 23 theme various 186:192 arg1 shape 225:229 cell shape 220:229 cell shape 220:229 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 23 theme various 186:192 arg1 function 237:244 cell function 232:244 cell function 232:244 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 2 23 theme various 186:192 arg1 adhesion 255:262 cell adhesion 250:262 cell adhesion 250:262 The composition and structure of the polysaccharides affect various cell properties including cell shape, cell function and cell adhesion. 28050839 3 24 dep PACE 265:268 arg1 using 295:299 using 295:299 using Carbohydrate gel Electrophoresis 295:332 PACE (Polysaccharide Analysis using Carbohydrate gel Electrophoresis) uses a simple, rapid technique to analyze polysaccharide quantity and structure (Goubet et al., Anal Biochem 300:53-68, 2002). 25498703 2 0 theme 1→3,6 370:374 arg1 -Galp 376:380 (1→3,6)-Galp 369:380 (1→3,6)-Galp 369:380 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 1 1 dep Bio-Gel 240:246 arg1 P2 248:249 P2 248:249 P2 248:249 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 2 2 dep -Galp 358:362 arg1 1→3 354:356 1→3 354:356 1→3 354:356 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 5 3 with O-6 1015:1017 arg1 arabinoses 1033:1042 terminal arabinoses 1024:1042 terminal arabinoses 1024:1042 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 3 4 with ELISA 607:611 arg1 assay 667:671 Yariv reagent assay 653:671 Yariv reagent assay 653:671 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 3 4 with ELISA 607:611 arg1 antibodies 638:647 specific monoclonal antibodies 618:647 specific monoclonal antibodies 618:647 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 1 5 theme size-exclusion 207:220 arg1 chromatography 222:235 size-exclusion chromatography 207:235 size-exclusion chromatography on Bio-Gel P2 and Superdex 75 207:265 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 5 6 theme side 917:920 arg1 chain 922:926 The arabinogalactan side chain 897:926 The arabinogalactan side chain 897:926 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 4 7 theme -L-arabinofuranose 774:791 arg1 residues 793:800 seven α-(1→5)-L-arabinofuranose residues 761:800 seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain 761:894 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 1 8 dep Da 113:114 arg1 4 110:110 4 110:110 4 110:110 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 4 9 theme terminal 846:853 arg1 arabinoses 855:864 terminal arabinoses 846:864 terminal arabinoses 846:864 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 1 10 theme branched 63:70 arg1 galactoarabinan 72:86 A highly branched galactoarabinan 54:86 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) 54:115 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 4 11 contain had 734:736 arg1 unit 721:724 the proposed repeating unit 698:724 the proposed repeating unit of DAG1 698:732 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 4 11 contain had 734:736 arg2 backbone 740:747 a backbone 738:747 a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain 738:894 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 2 12 theme relative 387:394 arg1 proportion 396:405 a relative proportion 385:405 a relative proportion of approximately 6:4:3:3:3 385:432 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 3 13 theme specific 618:625 arg1 antibodies 638:647 specific monoclonal antibodies 618:647 specific monoclonal antibodies 618:647 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 3 14 dep the 502:504 arg1 aid 506:508 aid 506:508 aid 506:508 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 3 15 theme monoclonal 627:636 arg1 antibodies 638:647 specific monoclonal antibodies 618:647 specific monoclonal antibodies 618:647 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 0 16 theme Structural 0:9 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of galactoarabinan 0:37 Structural analysis of galactoarabinan from duckweed. 25498703 5 17 theme -D-galactopyranose 955:972 arg1 residues 974:981 six β-(1→3)-D-galactopyranose residues 944:981 six β-(1→3)-D-galactopyranose residues 944:981 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 2 18 theme Methylation 268:278 arg1 analysis 280:287 Methylation analysis 268:287 Methylation analysis 268:287 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 2 19 theme 1→5 328:330 arg1 -Araf 332:336 (1→5)-Araf 327:336 (1→5)-Araf 327:336 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 0 20 theme galactoarabinan 23:37 arg1 analysis 11:18 Structural analysis 0:18 Structural analysis of galactoarabinan 0:37 Structural analysis of galactoarabinan from duckweed. 25498703 3 21 theme arabinan 513:520 arg1 enzymes 532:538 arabinan degrading enzymes 513:538 arabinan degrading enzymes 513:538 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 1 22 from chromatography 222:235 arg1 Bio-Gel 240:246 Bio-Gel P2 and Superdex 75 240:265 Bio-Gel 240:246 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 1 22 from chromatography 222:235 arg1 Superdex 255:262 Superdex 75 255:265 Superdex 75 255:265 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 1 23 theme Mw∼4.0×10 100:108 arg1 Da 113:114 Mw∼4.0×10(4) Da 100:114 Mw∼4.0×10(4) Da 100:114 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 1 23 theme Mw∼4.0×10 100:108 arg1 DAG1 94:97 DAG1 94:97 DAG1 (Mw∼4.0×10(4) Da) 94:115 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 1 24 theme % 167:167 arg1 extraction 183:192 70% (v/v) ethanol extraction 165:192 70% (v/v) ethanol extraction 165:192 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 3 25 theme Yariv 653:657 arg1 assay 667:671 Yariv reagent assay 653:671 Yariv reagent assay 653:671 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 5 26 theme residues 974:981 arg1 half 984:987 half 984:987 half 984:987 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 3 27 theme reagent 659:665 arg1 assay 667:671 Yariv reagent assay 653:671 Yariv reagent assay 653:671 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 2 28 theme 6:4:3:3:3 424:432 arg1 proportion 396:405 a relative proportion 385:405 a relative proportion of approximately 6:4:3:3:3 385:432 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 3 29 theme DAG1 558:561 arg1 unit 573:576 DAG1 repeating unit 558:576 DAG1 repeating unit 558:576 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 1 30 theme 70 165:166 arg1 % 167:167 % 167:167 % 167:167 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 2 31 theme 1→2,5 340:344 arg1 -Araf 346:350 (1→2,5)-Araf 339:350 (1→2,5)-Araf 339:350 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 4 32 theme DAG1 729:732 arg1 unit 721:724 the proposed repeating unit 698:724 the proposed repeating unit of DAG1 698:732 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 5 33 theme terminal 1024:1031 arg1 arabinoses 1033:1042 terminal arabinoses 1024:1042 terminal arabinoses 1024:1042 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 5 34 theme last 1052:1055 arg1 galactose 1057:1065 the last galactose 1048:1065 the last galactose 1048:1065 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 4 35 theme arabinogalactan 869:883 arg1 chain 890:894 arabinogalactan side chain 869:894 arabinogalactan side chain 869:894 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 2 36 theme arabinogalactan/galactoarabinan 449:479 arg1 polysacchairde 481:494 an arabinogalactan/galactoarabinan polysacchairde 446:494 an arabinogalactan/galactoarabinan polysacchairde 446:494 Methylation analysis showed that DAG1 consisted of t-Araf, (1→5)-Araf, (1→2,5)-Araf, (1→3)-Galp, and (1→3,6)-Galp in a relative proportion of approximately 6:4:3:3:3, suggesting an arabinogalactan/galactoarabinan polysacchairde. 25498703 5 37 theme arabinogalactan 901:915 arg1 chain 922:926 The arabinogalactan side chain 897:926 The arabinogalactan side chain 897:926 The arabinogalactan side chain was composed of six β-(1→3)-D-galactopyranose residues, half of which were ramified at O-6 with terminal arabinoses and the last galactose was terminated with arabinose. 25498703 4 38 theme repeating 711:719 arg1 unit 721:724 the proposed repeating unit 698:724 the proposed repeating unit of DAG1 698:732 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 1 39 theme ethanol 175:181 arg1 extraction 183:192 70% (v/v) ethanol extraction 165:192 70% (v/v) ethanol extraction 165:192 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 3 40 theme repeating 563:571 arg1 unit 573:576 DAG1 repeating unit 558:576 DAG1 repeating unit 558:576 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 4 41 theme proposed 702:709 arg1 unit 721:724 the proposed repeating unit 698:724 the proposed repeating unit of DAG1 698:732 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 3 42 theme unit 573:576 arg1 structure 545:553 the structure 541:553 the structure of DAG1 repeating unit 541:576 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 25498703 1 43 dep % 167:167 arg1 v/v 170:172 v/v 170:172 v/v 170:172 A highly branched galactoarabinan named DAG1 (Mw∼4.0×10(4) Da) was purified from Lemna aequinoctialis 6000 via 70% (v/v) ethanol extraction, followed by size-exclusion chromatography on Bio-Gel P2 and Superdex 75. 25498703 4 44 theme side 885:888 arg1 chain 890:894 arabinogalactan side chain 869:894 arabinogalactan side chain 869:894 Analyses indicated that the proposed repeating unit of DAG1 had a backbone composed of seven α-(1→5)-L-arabinofuranose residues where branching occurred at O-2 with either terminal arabinoses or arabinogalactan side chain. 25498703 3 45 theme degrading 522:530 arg1 enzymes 532:538 arabinan degrading enzymes 513:538 arabinan degrading enzymes 513:538 With the aid of arabinan degrading enzymes, the structure of DAG1 repeating unit was further characterized by ELISA with specific monoclonal antibodies and Yariv reagent assay. 28669498 0 0 theme proximity 82:90 arg1 ligation 92:99 in situ proximity ligation 74:99 in situ proximity ligation 74:99 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 2 1 from function 455:462 arg1 understanding 387:399 an in-depth understanding 375:399 an in-depth understanding of the role of glycosylation on protein structure and function 375:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 1 2 theme disease 279:285 arg1 progression 287:297 disease progression 279:297 disease progression 279:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 4 3 theme glycan 676:681 arg1 tagging 693:699 glycan metabolic tagging 676:699 glycan metabolic tagging 676:699 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 5 4 theme GalNAcylation 813:825 arg1 visualization 766:778 the visualization 762:778 the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins 762:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 7 5 theme epidermal 1086:1094 arg1 receptor 1110:1117 endogenous epidermal growth factor receptor 1075:1117 endogenous epidermal growth factor receptor 1075:1117 We further show that our strategy can be applied to image the dimerization of endogenous epidermal growth factor receptor. 28669498 6 6 theme proteins 987:994 arg1 fraction 971:978 the glycosylated fraction 954:978 the glycosylated fraction of the proteins 954:994 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 6 7 theme spatial 885:891 arg1 resolution 893:902 the high spatial resolution 876:902 the high spatial resolution of this method 876:917 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 2 8 theme glycosylation 312:324 arg1 status 326:331 the glycosylation status 308:331 the glycosylation status on a specific protein of interest 308:365 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 1 9 theme critical 192:199 arg1 transduction 262:273 signal transduction 255:273 signal transduction 255:273 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 9 theme critical 192:199 arg1 progression 287:297 disease progression 279:297 disease progression 279:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 9 theme critical 192:199 arg1 processes 212:220 critical biological processes 192:220 critical biological processes including cell-cell recognition, signal transduction and disease progression 192:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 9 theme critical 192:199 arg1 recognition 242:252 cell-cell recognition 232:252 cell-cell recognition 232:252 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 2 10 theme protein 433:439 arg1 structure 441:449 protein structure 433:449 protein structure 433:449 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 5 11 theme several 830:836 arg1 proteins 857:864 several important membrane proteins 830:864 several important membrane proteins 830:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 6 12 theme high 880:883 arg1 resolution 893:902 the high spatial resolution 876:902 the high spatial resolution of this method 876:917 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 4 13 theme metabolic 683:691 arg1 tagging 693:699 glycan metabolic tagging 676:699 glycan metabolic tagging 676:699 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 2 14 from understanding 387:399 arg1 function 455:462 function 455:462 function 455:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 2 14 from understanding 387:399 arg1 structure 441:449 protein structure 433:449 protein structure 433:449 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 2 15 theme glycosylation 416:428 arg1 role 408:411 the role 404:411 the role of glycosylation on protein structure and function 404:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 8 16 theme unique 1147:1152 arg1 tool 1154:1157 a unique tool 1145:1157 a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context 1145:1233 Thus, our study provides a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context. 28669498 2 17 from role 408:411 arg1 function 455:462 function 455:462 function 455:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 2 17 from role 408:411 arg1 structure 441:449 protein structure 433:449 protein structure 433:449 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 6 18 gly glycosylated 958:969 arg1 fraction 971:978 the glycosylated fraction 954:978 the glycosylated fraction of the proteins 954:994 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 1 19 theme biological 201:210 arg1 transduction 262:273 signal transduction 255:273 signal transduction 255:273 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 19 theme biological 201:210 arg1 progression 287:297 disease progression 279:297 disease progression 279:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 19 theme biological 201:210 arg1 processes 212:220 critical biological processes 192:220 critical biological processes including cell-cell recognition, signal transduction and disease progression 192:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 19 theme biological 201:210 arg1 recognition 242:252 cell-cell recognition 232:252 cell-cell recognition 232:252 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 0 20 theme glycosylation 28:40 arg1 Imaging 0:6 Imaging 0:6 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 0:100 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 2 21 theme specific 338:345 arg1 protein 347:353 a specific protein 336:353 a specific protein of interest 336:365 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 1 22 theme processes 212:220 arg1 transduction 262:273 signal transduction 255:273 signal transduction 255:273 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 22 theme processes 212:220 arg1 progression 287:297 disease progression 279:297 disease progression 279:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 22 theme processes 212:220 arg1 number 182:187 a number 180:187 a number of critical biological processes including cell-cell recognition, signal transduction and disease progression 180:297 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 22 theme processes 212:220 arg1 recognition 242:252 cell-cell recognition 232:252 cell-cell recognition 232:252 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 0 23 theme protein-specific 11:26 arg1 glycosylation 28:40 protein-specific glycosylation 11:40 protein-specific glycosylation 11:40 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 4 24 theme protein-specific 632:647 arg1 glycosylation 649:661 the protein-specific glycosylation 628:661 the protein-specific glycosylation 628:661 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 5 25 theme sialylation 783:793 arg1 visualization 766:778 the visualization 762:778 the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins 762:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 6 26 theme fraction 971:978 arg1 localization 938:949 subcellular localization 926:949 subcellular localization of the glycosylated fraction of the proteins 926:994 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 8 27 gly glycosylation 1191:1203 arg1 context 1227:1233 a dynamic cellular context 1208:1233 a dynamic cellular context 1208:1233 Thus, our study provides a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context. 28669498 0 28 theme glycan 45:50 arg1 tagging 62:68 glycan metabolic tagging 45:68 glycan metabolic tagging 45:68 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 8 29 theme protein-specific 1174:1189 arg1 glycosylation 1191:1203 the protein-specific glycosylation 1170:1203 the protein-specific glycosylation 1170:1203 Thus, our study provides a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context. 28669498 6 30 theme glycosylated 958:969 arg1 fraction 971:978 the glycosylated fraction 954:978 the glycosylated fraction of the proteins 954:994 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 1 31 theme cell-cell 232:240 arg1 recognition 242:252 cell-cell recognition 232:252 cell-cell recognition 232:252 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 5 32 theme important 838:846 arg1 proteins 857:864 several important membrane proteins 830:864 several important membrane proteins 830:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 4 33 theme fluorescence 585:596 arg1 strategy 606:613 a highly sensitive fluorescence imaging strategy 566:613 a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA) 566:744 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 5 34 theme fucosylation 796:807 arg1 visualization 766:778 the visualization 762:778 the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins 762:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 7 35 theme receptor 1110:1117 arg1 dimerization 1059:1070 the dimerization 1055:1070 the dimerization of endogenous epidermal growth factor receptor 1055:1117 We further show that our strategy can be applied to image the dimerization of endogenous epidermal growth factor receptor. 28669498 3 36 theme protein-specific 497:512 arg1 glycosylation 514:526 protein-specific glycosylation 497:526 protein-specific glycosylation 497:526 However, methods for monitoring protein-specific glycosylation are largely lacking. 28669498 4 37 theme sensitive 575:583 arg1 strategy 606:613 a highly sensitive fluorescence imaging strategy 566:613 a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA) 566:744 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 8 38 theme cellular 1218:1225 arg1 context 1227:1233 a dynamic cellular context 1208:1233 a dynamic cellular context 1208:1233 Thus, our study provides a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context. 28669498 5 39 from visualization 766:778 arg1 proteins 857:864 several important membrane proteins 830:864 several important membrane proteins 830:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 2 40 theme in-depth 378:385 arg1 understanding 387:399 an in-depth understanding 375:399 an in-depth understanding of the role of glycosylation on protein structure and function 375:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 0 41 theme metabolic 52:60 arg1 tagging 62:68 glycan metabolic tagging 45:68 glycan metabolic tagging 45:68 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 4 42 dep in 705:706 arg1 situ 708:711 situ 708:711 situ 708:711 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 7 43 theme growth 1096:1101 arg1 receptor 1110:1117 endogenous epidermal growth factor receptor 1075:1117 endogenous epidermal growth factor receptor 1075:1117 We further show that our strategy can be applied to image the dimerization of endogenous epidermal growth factor receptor. 28669498 1 44 theme important 122:130 arg1 modification 150:161 an important posttranslational modification 119:161 an important posttranslational modification 119:161 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 44 theme important 122:130 arg1 Glycosylation 102:114 Glycosylation 102:114 Glycosylation 102:114 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 4 45 theme imaging 598:604 arg1 strategy 606:613 a highly sensitive fluorescence imaging strategy 566:613 a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA) 566:744 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 8 46 theme dynamic 1210:1216 arg1 context 1227:1233 a dynamic cellular context 1208:1233 a dynamic cellular context 1208:1233 Thus, our study provides a unique tool to monitor the protein-specific glycosylation in a dynamic cellular context. 28669498 6 47 theme subcellular 926:936 arg1 localization 938:949 subcellular localization 926:949 subcellular localization of the glycosylated fraction of the proteins 926:994 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 7 48 theme factor 1103:1108 arg1 receptor 1110:1117 endogenous epidermal growth factor receptor 1075:1117 endogenous epidermal growth factor receptor 1075:1117 We further show that our strategy can be applied to image the dimerization of endogenous epidermal growth factor receptor. 28669498 1 49 theme posttranslational 132:148 arg1 modification 150:161 an important posttranslational modification 119:161 an important posttranslational modification 119:161 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 1 49 theme posttranslational 132:148 arg1 Glycosylation 102:114 Glycosylation 102:114 Glycosylation 102:114 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 0 50 dep in 74:75 arg1 situ 77:80 situ 77:80 situ 77:80 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 4 51 dep tagging 693:699 arg1 termed 733:738 termed 733:738 termed GPLA 733:743 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 7 52 theme endogenous 1075:1084 arg1 receptor 1110:1117 endogenous epidermal growth factor receptor 1075:1117 endogenous epidermal growth factor receptor 1075:1117 We further show that our strategy can be applied to image the dimerization of endogenous epidermal growth factor receptor. 28669498 2 53 theme interest 358:365 arg1 protein 347:353 a specific protein 336:353 a specific protein of interest 336:365 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 1 54 theme signal 255:260 arg1 transduction 262:273 signal transduction 255:273 signal transduction 255:273 Glycosylation is an important posttranslational modification, which regulates a number of critical biological processes including cell-cell recognition, signal transduction and disease progression. 28669498 0 55 theme in 74:75 arg1 ligation 92:99 in situ proximity ligation 74:99 in situ proximity ligation 74:99 Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. 28669498 6 56 theme method 912:917 arg1 resolution 893:902 the high spatial resolution 876:902 the high spatial resolution of this method 876:917 Notably, the high spatial resolution of this method allows subcellular localization of the glycosylated fraction of the proteins. 28669498 2 57 from structure 441:449 arg1 understanding 387:399 an in-depth understanding 375:399 an in-depth understanding of the role of glycosylation on protein structure and function 375:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 2 58 theme role 408:411 arg1 understanding 387:399 an in-depth understanding 375:399 an in-depth understanding of the role of glycosylation on protein structure and function 375:462 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 2 59 from status 326:331 arg1 protein 347:353 a specific protein 336:353 a specific protein of interest 336:365 Probing the glycosylation status on a specific protein of interest enables an in-depth understanding of the role of glycosylation on protein structure and function. 28669498 4 60 theme in 705:706 arg1 ligation 723:730 in situ proximity ligation 705:730 in situ proximity ligation 705:730 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28669498 5 61 theme membrane 848:855 arg1 proteins 857:864 several important membrane proteins 830:864 several important membrane proteins 830:864 We demonstrate the visualization of sialylation, fucosylation and GalNAcylation on several important membrane proteins. 28669498 4 62 theme proximity 713:721 arg1 ligation 723:730 in situ proximity ligation 705:730 in situ proximity ligation 705:730 Here we describe a highly sensitive fluorescence imaging strategy to visualize the protein-specific glycosylation by combining glycan metabolic tagging and in situ proximity ligation (termed GPLA). 28575723 5 0 theme structural 961:970 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 0 theme structural 961:970 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 1 gly N-glycosylation 342:356 arg2 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 3 2 attach linked 403:408 arg2 -glycans 394:401 N -glycans 392:401 N -glycans linked to Cha o 3 392:419 However, the structures of N -glycans linked to Cha o 3 remain to be determined. 28575723 3 2 attach linked 403:408 arg1 o 417:417 Cha o 3 413:419 Cha o 3 413:419 However, the structures of N -glycans linked to Cha o 3 remain to be determined. 28575723 0 3 dep cypress 94:100 arg1 Hinoki 103:108 Hinoki 103:108 Hinoki 103:108 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 1 4 theme cypress 195:201 arg1 pollen 204:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 5 5 from pollens 1016:1022 arg1 glycoallergens 983:996 glycoallergens 983:996 glycoallergens from Cupressaceae pollens 983:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 5 from pollens 1016:1022 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 5 from pollens 1016:1022 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 6 theme Lea 774:776 arg1 epitope 778:784 the Lea epitope 770:784 the Lea epitope 770:784 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 7 theme acid 230:233 arg1 sequence 235:242 The deduced amino acid sequence 212:242 The deduced amino acid sequence of Cha o 3 212:253 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 5 8 theme Cry 809:811 arg1 j1 813:814 Cry j1 809:814 Cry j1 809:814 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 9 theme amino 224:228 arg1 sequence 235:242 The deduced amino acid sequence 212:242 The deduced amino acid sequence of Cha o 3 212:253 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 10 contain contains 289:296 arg2 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 10 contain contains 289:296 arg2 domain 310:315 a cellulase domain 298:315 a cellulase domain 298:315 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 10 contain contains 289:296 arg2 number 323:328 a number 321:328 a number of putative N-glycosylation sites 321:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 10 contain contains 289:296 arg1 glycoallergen 275:287 this glycoallergen 270:287 this glycoallergen 270:287 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 1 11 gly glycosylated 137:148 arg1 allergen 150:157 a newly found glycosylated allergen 123:157 a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen 123:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 1 11 gly glycosylated 137:148 arg1 o 116:116 Cha o 3 112:118 Cha o 3 112:118 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 2 12 theme N-glycosylation 342:356 arg1 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 13 theme deduced 216:222 arg1 sequence 235:242 The deduced amino acid sequence 212:242 The deduced amino acid sequence of Cha o 3 212:253 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 5 14 theme Cha 754:756 arg1 o 758:758 Cha o 3 754:760 Cha o 3 754:760 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 15 from feature 972:978 arg1 pollens 1016:1022 Cupressaceae pollens 1003:1022 Cupressaceae pollens 1003:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 4 16 theme major 593:597 arg1 structures 599:608 major structures 593:608 major structures 593:608 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 5 17 theme o 758:758 arg1 glycoform 741:749 The glycoform 737:749 The glycoform of Cha o 3 bearing the Lea epitope 737:784 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 17 theme o 758:758 arg1 similar 789:795 similar 789:795 similar 789:795 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 18 theme o 251:251 arg1 sequence 235:242 The deduced amino acid sequence 212:242 The deduced amino acid sequence of Cha o 3 212:253 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 5 19 theme GlcNAc2Man3Xyl1Fuc1GlcNAc2 922:947 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 19 theme GlcNAc2Man3Xyl1Fuc1GlcNAc2 922:947 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 4 20 theme plant 563:567 arg1 N-glycans 582:590 plant complex-type N-glycans 563:590 plant complex-type N-glycans 563:590 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 5 21 gly glycoform 741:749 arg1 o 758:758 Cha o 3 754:760 Cha o 3 754:760 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 22 theme Cha 247:249 arg1 o 251:251 Cha o 3 247:253 Cha o 3 247:253 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 3 23 theme -glycans 394:401 arg1 structures 378:387 the structures 374:387 the structures of N -glycans linked to Cha o 3 374:419 However, the structures of N -glycans linked to Cha o 3 remain to be determined. 28575723 1 24 from pollen 204:209 arg1 allergen 150:157 a newly found glycosylated allergen 123:157 a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen 123:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 1 24 from pollen 204:209 arg1 o 116:116 Cha o 3 112:118 Cha o 3 112:118 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 4 25 gly glycoform 488:496 arg1 o 505:505 Cha o 3 501:507 Cha o 3 501:507 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 1 26 theme found 131:135 arg1 allergen 150:157 a newly found glycosylated allergen 123:157 a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen 123:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 1 26 theme found 131:135 arg1 o 116:116 Cha o 3 112:118 Cha o 3 112:118 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 3 27 theme N 392:392 arg1 -glycans 394:401 N -glycans 392:401 N -glycans linked to Cha o 3 392:419 However, the structures of N -glycans linked to Cha o 3 remain to be determined. 28575723 0 28 theme Japanese 85:92 arg1 obtusa 77:82 obtusa 77:82 obtusa 77:82 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 0 28 theme Japanese 85:92 arg1 cypress 94:100 Japanese cypress 85:100 Japanese cypress 85:100 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 5 29 theme Jun 817:819 arg1 a 821:821 Jun a 1 817:823 Jun a 1 817:823 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 1 30 theme glycosylated 137:148 arg1 allergen 150:157 a newly found glycosylated allergen 123:157 a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen 123:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 1 30 theme glycosylated 137:148 arg1 o 116:116 Cha o 3 112:118 Cha o 3 112:118 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 0 31 theme pollen 32:37 arg1 allergen 39:46 a newly identified pollen allergen 13:46 a newly identified pollen allergen 13:46 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 0 31 theme pollen 32:37 arg1 o 53:53 Cha o 3 49:55 Cha o 3 49:55 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 5 32 theme major 838:842 arg1 glycoallergens 844:857 major glycoallergens 838:857 major glycoallergens in cedar or cypress pollens 838:885 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 32 theme major 838:842 arg1 a 833:833 Cup a 1 829:835 Cup a 1 829:835 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 33 from glycoallergens 844:857 arg1 pollens 879:885 cypress pollens 871:885 cypress pollens 871:885 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 33 from glycoallergens 844:857 arg1 cedar 862:866 cedar 862:866 cedar 862:866 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 2 34 theme putative 333:340 arg1 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 0 35 theme identified 21:30 arg1 allergen 39:46 a newly identified pollen allergen 13:46 a newly identified pollen allergen 13:46 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 0 35 theme identified 21:30 arg1 o 53:53 Cha o 3 49:55 Cha o 3 49:55 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 5 36 theme glycoallergens 983:996 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 36 theme glycoallergens 983:996 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 37 theme predominant 896:906 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 37 theme predominant 896:906 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 3 38 theme Cha 413:415 arg1 o 417:417 Cha o 3 413:419 Cha o 3 413:419 However, the structures of N -glycans linked to Cha o 3 remain to be determined. 28575723 0 39 theme allergen 39:46 arg1 Glycoform 0:8 Glycoform 0:8 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 0:110 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 5 40 theme Cupressaceae 1003:1014 arg1 pollens 1016:1022 Cupressaceae pollens 1003:1022 Cupressaceae pollens 1003:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 41 theme common 954:959 arg1 feature 972:978 a common structural feature 952:978 a common structural feature of glycoallergens from Cupressaceae pollens 952:1022 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 41 theme common 954:959 arg1 occurrence 908:917 the predominant occurrence 892:917 the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 892:947 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 42 theme cypress 871:877 arg1 pollens 879:885 cypress pollens 871:885 cypress pollens 871:885 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 0 43 from obtusa 77:82 arg1 Glycoform 0:8 Glycoform 0:8 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 0:110 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 5 44 theme Cup 829:831 arg1 glycoallergens 844:857 major glycoallergens 838:857 major glycoallergens in cedar or cypress pollens 838:885 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 5 44 theme Cup 829:831 arg1 a 833:833 Cup a 1 829:835 Cup a 1 829:835 The glycoform of Cha o 3 bearing the Lea epitope is similar to those of Cry j1, Jun a 1, or Cup a 1, major glycoallergens in cedar or cypress pollens, and the predominant occurrence of GlcNAc2Man3Xyl1Fuc1GlcNAc2 is a common structural feature of glycoallergens from Cupressaceae pollens. 28575723 1 45 theme Chamaecyparis 164:176 arg1 pollen 204:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 4 46 theme complex-type 569:580 arg1 N-glycans 582:590 plant complex-type N-glycans 563:590 plant complex-type N-glycans 563:590 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 1 47 theme obtusa 178:183 arg1 pollen 204:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 4 48 contain carries 543:549 arg2 N-glycans 582:590 plant complex-type N-glycans 563:590 plant complex-type N-glycans 563:590 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 4 48 contain carries 543:549 arg1 glycoallergen 529:541 this glycoallergen 524:541 this glycoallergen 524:541 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 2 49 theme sites 358:362 arg1 domain 310:315 a cellulase domain 298:315 a cellulase domain 298:315 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 49 theme sites 358:362 arg1 number 323:328 a number 321:328 a number of putative N-glycosylation sites 321:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 49 theme sites 358:362 arg1 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 50 theme cellulase 300:308 arg1 domain 310:315 a cellulase domain 298:315 a cellulase domain 298:315 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 2 50 theme cellulase 300:308 arg1 sites 358:362 putative N-glycosylation sites 333:362 putative N-glycosylation sites 333:362 The deduced amino acid sequence of Cha o 3 indicates that this glycoallergen contains a cellulase domain and a number of putative N-glycosylation sites. 28575723 0 51 theme Cha 49:51 arg1 allergen 39:46 a newly identified pollen allergen 13:46 a newly identified pollen allergen 13:46 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 0 51 theme Cha 49:51 arg1 o 53:53 Cha o 3 49:55 Cha o 3 49:55 Glycoform of a newly identified pollen allergen, Cha o 3, from Chamaecyparis obtusa (Japanese cypress, Hinoki). 28575723 1 52 theme Cha 112:114 arg1 allergen 150:157 a newly found glycosylated allergen 123:157 a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen 123:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 1 52 theme Cha 112:114 arg1 o 116:116 Cha o 3 112:118 Cha o 3 112:118 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 28575723 4 53 theme o 505:505 arg1 glycoform 488:496 the glycoform 484:496 the glycoform of Cha o 3 484:507 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 4 54 theme Cha 501:503 arg1 o 505:505 Cha o 3 501:507 Cha o 3 501:507 In this study, therefore, we analyzed the glycoform of Cha o 3 and found that this glycoallergen carries exclusively plant complex-type N-glycans; major structures were GlcNAc2Man3Xyl1Fuc1GlcNAc2 (39%), Gal1Fuc1GlcNAc2Man3Xyl1Fuc1GlcNAc2 (14%), and Gal2Fuc2GlcNAc2Man3Xyl1Fuc1GlcNAc2 (25%). 28575723 1 55 theme Japanese 186:193 arg1 pollen 204:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Chamaecyparis obtusa (Japanese cypress) pollen 164:209 Cha o 3 is a newly found glycosylated allergen from Chamaecyparis obtusa (Japanese cypress) pollen. 27765135 4 0 theme Plasma 547:552 arg1 N-glycome 558:566 Plasma IgG N-glycome 547:566 Plasma IgG N-glycome 547:566 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 2 1 from Hospital 397:404 arg1 participants 364:375 669 participants 360:375 669 participants from Beijing Xuanwu Hospital during January-June 2012 360:429 Methods: A cluster sampling was conducted to randomly select 669 participants from Beijing Xuanwu Hospital during January-June 2012. 27765135 8 2 theme participants 1019:1030 arg1 total 998:1002 A total 996:1002 A total of 669 healthy participants 996:1030 Results: A total of 669 healthy participants were included, including 235 men (35.13%) and 434 women (64.87%). 27765135 9 3 theme participants 1121:1132 arg1 age 1110:1112 The average age 1098:1112 The average age of the participants 1098:1132 The average age of the participants was (44.23±9.82) years. 27765135 9 3 theme participants 1121:1132 arg1 years 1151:1155 (44.23±9.82) years 1138:1155 (44.23±9.82) years 1138:1155 The average age of the participants was (44.23±9.82) years. 27765135 4 4 theme mass 683:686 arg1 spectrometry 688:699 mass spectrometry 683:699 mass spectrometry 683:699 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 7 5 theme nonparametric 879:891 arg1 test 893:896 nonparametric test 879:896 nonparametric test 879:896 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 4 6 theme IgG 554:556 arg1 N-glycome 558:566 Plasma IgG N-glycome 547:566 Plasma IgG N-glycome 547:566 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 10 7 from level 1516:1520 arg1 males 1525:1529 males 1525:1529 males 1525:1529 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 7 from level 1516:1520 arg1 females 1539:1545 females 1539:1545 females 1539:1545 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 3 8 theme pressure 516:523 arg1 tube 525:528 vacuum negative pressure tube 500:528 vacuum negative pressure tube containing EDTA 500:544 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 4 9 theme BEH 646:648 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 10 10 dep Z=-2.343 1312:1319 arg1 P=0.019 1322:1328 P=0.019 1322:1328 P=0.019 1322:1328 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 1 11 theme structure 249:257 arg1 distribution 229:240 the distribution 225:240 the distribution of the structure of IgG N-glycans in general population 225:296 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 12 theme saliva 1495:1500 arg1 level 1516:1520 a higher saliva acidification level 1486:1520 a higher saliva acidification level in males than in females 1486:1545 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 3 13 theme vacuum 500:505 arg1 tube 525:528 vacuum negative pressure tube 500:528 vacuum negative pressure tube containing EDTA 500:544 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 10 14 theme higher 1232:1237 arg1 level 1210:1214 the galactosylation level 1190:1214 the galactosylation level 1190:1214 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 14 theme higher 1232:1237 arg1 Z=-2.192 1263:1270 Z=-2.192 1263:1270 Z=-2.192 1263:1270 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 14 theme higher 1232:1237 arg1 [Fn 1259:1261 higher than those of males [Fn 1232:1261 higher than those of males [Fn(Z=-2.192, P=0.028) 1232:1280 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 1 15 theme immunoglobulin 158:171 arg1 IgG 175:177 IgG 175:177 IgG 175:177 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 1 15 theme immunoglobulin 158:171 arg1 G 173:173 immunoglobulin G 158:173 immunoglobulin G(IgG) N-glycans 158:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 16 theme higher 1488:1493 arg1 level 1516:1520 a higher saliva acidification level 1486:1520 a higher saliva acidification level in males than in females 1486:1545 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 11 17 theme N-glycosylation 1564:1578 arg1 levels 1580:1585 IgG N-glycosylation levels 1560:1585 IgG N-glycosylation levels 1560:1585 Conclusion: IgG N-glycosylation levels might be gender dependent. 27765135 3 18 theme negative 507:514 arg1 tube 525:528 vacuum negative pressure tube 500:528 vacuum negative pressure tube containing EDTA 500:544 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 7 19 theme Independent 850:860 arg1 t-test 869:874 Independent sample t-test 850:874 Independent sample t-test 850:874 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 1 20 theme G 173:173 arg1 N-glycans 180:188 immunoglobulin G(IgG) N-glycans 158:188 immunoglobulin G(IgG) N-glycans 158:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 21 theme females 1434:1440 arg1 FS1/FS2 1403:1409 FS1/FS2 1403:1409 FS1/FS2 (Z=-3.336, P=0.001) of females 1403:1440 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 8 22 theme healthy 1011:1017 arg1 participants 1019:1030 669 healthy participants 1007:1030 669 healthy participants 1007:1030 Results: A total of 669 healthy participants were included, including 235 men (35.13%) and 434 women (64.87%). 27765135 4 23 theme Waters 639:644 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 7 24 theme sample 862:867 arg1 t-test 869:874 Independent sample t-test 850:874 Independent sample t-test 850:874 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 4 25 theme chromatography 657:670 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 10 26 dep G2n 1332:1334 arg1 P<0.001 1347:1353 P<0.001 1347:1353 P<0.001 1347:1353 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 26 dep G2n 1332:1334 arg1 Z=-8.414 1337:1344 Z=-8.414 1337:1344 Z=-8.414 1337:1344 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 7 27 theme specific 930:937 arg1 difference 939:948 the gender specific difference 919:948 the gender specific difference 919:948 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 10 28 dep Z=-2.192 1263:1270 arg1 P=0.028 1273:1279 P=0.028 1273:1279 P=0.028 1273:1279 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 4 29 theme Glycan 650:655 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 0 30 theme specific 22:29 arg1 profiles 41:48 gender specific structure profiles 15:48 gender specific structure profiles of immunoglobulin G N-glycans 15:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 3 31 contain containing 530:539 arg2 EDTA 541:544 EDTA 541:544 EDTA 541:544 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 3 31 contain containing 530:539 arg1 tube 525:528 vacuum negative pressure tube 500:528 vacuum negative pressure tube containing EDTA 500:544 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 10 32 dep level 1180:1184 arg1 The 1158:1160 The 1158:1160 The 1158:1160 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 33 dep FtotalS1/FtotalS2 1362:1378 arg1 P<0.001 1390:1396 P<0.001 1390:1396 P<0.001 1390:1396 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 33 dep FtotalS1/FtotalS2 1362:1378 arg1 Z=-5.049 1380:1387 Z=-5.049 1380:1387 Z=-5.049 1380:1387 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 34 theme acidification 1502:1514 arg1 level 1516:1520 a higher saliva acidification level 1486:1520 a higher saliva acidification level in males than in females 1486:1545 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 0 35 theme gender 15:20 arg1 profiles 41:48 gender specific structure profiles 15:48 gender specific structure profiles of immunoglobulin G N-glycans 15:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 1 36 theme N-glycans 266:274 arg1 structure 249:257 the structure 245:257 the structure of IgG N-glycans 245:274 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 37 dep FS1/FS2 1403:1409 arg1 P=0.001 1422:1428 P=0.001 1422:1428 P=0.001 1422:1428 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 37 dep FS1/FS2 1403:1409 arg1 Z=-3.336 1412:1419 Z=-3.336 1412:1419 Z=-3.336 1412:1419 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 1 38 theme N-glycans 180:188 arg1 profiles 146:153 the gender specific structure profiles 116:153 the gender specific structure profiles of immunoglobulin G(IgG) N-glycans 116:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 0 39 theme profiles 41:48 arg1 [Comparison 0:10 [Comparison 0:10 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans 0:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 1 40 from distribution 229:240 arg1 population 287:296 general population 279:296 general population 279:296 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 41 theme galactosylation 1194:1208 arg1 level 1210:1214 the galactosylation level 1190:1214 the galactosylation level 1190:1214 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 41 theme galactosylation 1194:1208 arg1 [Fn 1259:1261 higher than those of males [Fn 1232:1261 higher than those of males [Fn(Z=-2.192, P=0.028) 1232:1280 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 42 theme core 1162:1165 arg1 fucosylation 1167:1178 core fucosylation 1162:1178 core fucosylation level 1162:1184 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 3 43 theme blood 439:443 arg1 sample 445:450 Venous blood sample 432:450 Venous blood sample (5 ml) 432:457 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 3 43 theme blood 439:443 arg1 ml 455:456 5 ml 453:456 5 ml 453:456 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 0 44 theme structure 31:39 arg1 profiles 41:48 gender specific structure profiles 15:48 gender specific structure profiles of immunoglobulin G N-glycans 15:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 1 45 theme general 279:285 arg1 population 287:296 general population 279:296 general population 279:296 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 5 46 theme chromatographic 769:783 arg1 peaks 785:789 24 chromatographic peaks 766:789 24 chromatographic peaks 766:789 Finally, we got 24 chromatographic peaks. 27765135 4 47 from chromatography 614:627 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 0 48 theme immunoglobulin 53:66 arg1 G 68:68 immunoglobulin G 53:68 immunoglobulin G N-glycans 53:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 1 49 theme IgG 262:264 arg1 N-glycans 266:274 IgG N-glycans 262:274 IgG N-glycans 262:274 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 2 50 theme cluster 310:316 arg1 sampling 318:325 A cluster sampling 308:325 A cluster sampling 308:325 Methods: A cluster sampling was conducted to randomly select 669 participants from Beijing Xuanwu Hospital during January-June 2012. 27765135 10 51 dep G0n 1283:1285 arg1 P<0.001 1297:1303 P<0.001 1297:1303 P<0.001 1297:1303 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 51 dep G0n 1283:1285 arg1 Z=-7.898 1287:1294 Z=-7.898 1287:1294 Z=-7.898 1287:1294 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 7 52 theme gender 923:928 arg1 difference 939:948 the gender specific difference 919:948 the gender specific difference 919:948 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 4 53 theme ultra-performance 589:605 arg1 UPLC 630:633 UPLC 630:633 UPLC 630:633 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 4 53 theme ultra-performance 589:605 arg1 chromatography 614:627 ultra-performance liquid chromatography 589:627 ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column 589:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 6 54 theme carbohydrate 826:837 arg1 structure 839:847 a different carbohydrate structure 814:847 a different carbohydrate structure 814:847 Each peak represented a different carbohydrate structure. 27765135 10 55 theme females 1219:1225 arg1 level 1180:1184 core fucosylation level 1162:1184 core fucosylation level 1162:1184 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 55 theme females 1219:1225 arg1 level 1210:1214 the galactosylation level 1190:1214 the galactosylation level 1190:1214 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 10 55 theme females 1219:1225 arg1 [Fn 1259:1261 higher than those of males [Fn 1232:1261 higher than those of males [Fn(Z=-2.192, P=0.028) 1232:1280 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 9 56 theme average 1102:1108 arg1 age 1110:1112 The average age 1098:1112 The average age of the participants 1098:1132 The average age of the participants was (44.23±9.82) years. 27765135 9 56 theme average 1102:1108 arg1 years 1151:1155 (44.23±9.82) years 1138:1155 (44.23±9.82) years 1138:1155 The average age of the participants was (44.23±9.82) years. 27765135 1 57 theme gender 120:125 arg1 profiles 146:153 the gender specific structure profiles 116:153 the gender specific structure profiles of immunoglobulin G(IgG) N-glycans 116:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 10 58 theme fucosylation 1167:1178 arg1 level 1180:1184 core fucosylation level 1162:1184 core fucosylation level 1162:1184 The core fucosylation level and the galactosylation level of females were higher than those of males [Fn(Z=-2.192, P=0.028), G0n(Z=-7.898, P<0.001), G1n (Z=-2.343, P=0.019), G2n (Z=-8.414, P<0.001)], but FtotalS1/FtotalS2(Z=-5.049, P<0.001) and FS1/FS2 (Z=-3.336, P=0.001) of females were higher than those of males, indicating a higher saliva acidification level in males than in females. 27765135 0 59 theme N-glycans 70:78 arg1 profiles 41:48 gender specific structure profiles 15:48 gender specific structure profiles of immunoglobulin G N-glycans 15:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 6 60 theme different 816:824 arg1 structure 839:847 a different carbohydrate structure 814:847 a different carbohydrate structure 814:847 Each peak represented a different carbohydrate structure. 27765135 1 61 theme specific 127:134 arg1 profiles 146:153 the gender specific structure profiles 116:153 the gender specific structure profiles of immunoglobulin G(IgG) N-glycans 116:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 3 62 theme Venous 432:437 arg1 sample 445:450 Venous blood sample 432:450 Venous blood sample (5 ml) 432:457 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 3 62 theme Venous 432:437 arg1 ml 455:456 5 ml 453:456 5 ml 453:456 Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. 27765135 0 63 theme G 68:68 arg1 N-glycans 70:78 immunoglobulin G N-glycans 53:78 immunoglobulin G N-glycans 53:78 [Comparison of gender specific structure profiles of immunoglobulin G N-glycans]. 27765135 11 64 theme IgG 1560:1562 arg1 levels 1580:1585 IgG N-glycosylation levels 1560:1585 IgG N-glycosylation levels 1560:1585 Conclusion: IgG N-glycosylation levels might be gender dependent. 27765135 7 65 used used 903:906 arg2 t-test 869:874 Independent sample t-test 850:874 Independent sample t-test 850:874 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 7 65 used used 903:906 arg2 test 893:896 nonparametric test 879:896 nonparametric test 879:896 Independent sample t-test or nonparametric test were used to compare the gender specific difference, P<0.05 was regarded as significant. 27765135 1 66 theme structure 136:144 arg1 profiles 146:153 the gender specific structure profiles 116:153 the gender specific structure profiles of immunoglobulin G(IgG) N-glycans 116:188 Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. 27765135 4 67 from spectrometry 688:699 arg1 column 672:677 Waters BEH Glycan chromatography column 639:677 Waters BEH Glycan chromatography column 639:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 4 68 theme liquid 607:612 arg1 UPLC 630:633 UPLC 630:633 UPLC 630:633 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 27765135 4 68 theme liquid 607:612 arg1 chromatography 614:627 ultra-performance liquid chromatography 589:627 ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column 589:677 Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling. 25234305 8 0 theme EOC 1605:1607 arg1 patients 1609:1616 EOC patients 1605:1616 EOC patients 1605:1616 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 9 1 theme study 1646:1650 arg1 results 1623:1629 The results 1619:1629 The results of the current study 1619:1650 The results of the current study provide an improved dataset to be used in glycan biomarker discovery. 25234305 4 2 theme most 727:730 arg1 forms 732:736 most forms 727:736 most forms of cancer 727:746 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 8 3 theme tetraantennary 1496:1509 arg1 structures 1511:1520 tetraantennary structures 1496:1520 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 7 4 theme Reduced 1196:1202 arg1 levels 1204:1209 Reduced levels 1196:1209 Reduced levels of diantennary structures and of high-mannose 5 1196:1257 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 3 5 theme diseases 714:721 arg1 course 700:705 the course 696:705 the course of the diseases 696:721 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 3 6 theme total 681:685 arg1 serum 687:691 total serum 681:691 total serum 681:691 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 5 7 theme structures 1059:1068 arg1 assignment 1039:1048 the assignment 1035:1048 the assignment of novel structures 1035:1068 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 0 8 theme laser-induced 95:107 arg1 fluorescence 109:120 laser-induced fluorescence 95:120 laser-induced fluorescence 95:120 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 0 9 from Identification 0:13 arg1 serum 47:51 human serum 41:51 human serum 41:51 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 5 10 theme exoglycosidase 954:967 arg1 digestions 969:978 exoglycosidase digestions 954:978 exoglycosidase digestions 954:978 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 7 11 theme high-mannose 1244:1255 arg1 levels 1204:1209 Reduced levels 1196:1209 Reduced levels of diantennary structures and of high-mannose 5 1196:1257 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 0 12 from serum 47:51 arg1 Identification 0:13 Identification 0:13 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence 0:120 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 8 13 gly core-fucosylated 1531:1546 arg1 N-glycans 1563:1571 core-fucosylated tetraantennary N-glycans 1531:1571 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 7 14 theme structures 1226:1235 arg1 levels 1204:1209 Reduced levels 1196:1209 Reduced levels of diantennary structures and of high-mannose 5 1196:1257 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 6 15 theme ovarian 1165:1171 arg1 EOC 1190:1192 EOC 1190:1192 EOC 1190:1192 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 6 15 theme ovarian 1165:1171 arg1 patients 1180:1187 primary epithelial ovarian cancer patients 1146:1187 primary epithelial ovarian cancer patients (EOC) 1146:1193 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 4 16 theme cancer 741:746 arg1 forms 732:736 most forms 727:736 most forms of cancer 727:746 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 8 17 gly fucosylation 1463:1474 arg1 patients 1609:1616 EOC patients 1605:1616 EOC patients 1605:1616 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 1 18 from Alterations 167:177 arg1 glycosylation 182:194 glycosylation 182:194 glycosylation 182:194 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 3 19 theme structures 575:584 arg1 maximum 554:560 only a maximum 547:560 only a maximum of 12 glycan structures, the most abundant ones, 547:609 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 2 20 theme desialylated 437:448 arg1 N-glycans 450:458 desialylated N-glycans 437:458 desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis 437:535 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 5 21 gly glycoproteins 1000:1012 arg1 glycoproteins 1000:1012 standard glycoproteins 991:1012 standard glycoproteins 991:1012 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 2 22 gly desialylated 437:448 arg1 N-glycans 450:458 desialylated N-glycans 437:458 desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis 437:535 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 7 23 theme EOC 1297:1299 arg1 samples 1301:1307 the EOC samples 1293:1307 the EOC samples 1293:1307 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 6 24 theme primary 1146:1152 arg1 EOC 1190:1192 EOC 1190:1192 EOC 1190:1192 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 6 24 theme primary 1146:1152 arg1 patients 1180:1187 primary epithelial ovarian cancer patients 1146:1187 primary epithelial ovarian cancer patients (EOC) 1146:1193 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 8 25 from relevance 1483:1491 arg1 N-glycans 1563:1571 core-fucosylated tetraantennary N-glycans 1531:1571 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 8 25 from relevance 1483:1491 arg1 structures 1511:1520 tetraantennary structures 1496:1520 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 0 26 from isomers 30:36 arg1 serum 47:51 human serum 41:51 human serum 41:51 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 8 27 theme core-fucosylated 1531:1546 arg1 N-glycans 1563:1571 core-fucosylated tetraantennary N-glycans 1531:1571 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 5 28 theme healthy 928:934 arg1 serum 942:946 healthy human serum 928:946 healthy human serum using exoglycosidase digestions 928:978 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 3 29 from modulations 666:676 arg1 course 700:705 the course 696:705 the course of the diseases 696:721 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 1 30 theme relevant 298:305 arg1 information 318:328 relevant diagnostic information 298:328 relevant diagnostic information 298:328 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 1 30 theme relevant 298:305 arg1 changes 251:257 specific changes 242:257 specific changes in glycosylation 242:274 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 5 31 theme human 936:940 arg1 serum 942:946 healthy human serum 928:946 healthy human serum using exoglycosidase digestions 928:978 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 0 32 theme N-glycan 21:28 arg1 isomers 30:36 34 N-glycan isomers 18:36 34 N-glycan isomers in human serum 18:51 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 1 33 theme diagnostic 307:316 arg1 information 318:328 relevant diagnostic information 298:328 relevant diagnostic information 298:328 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 1 33 theme diagnostic 307:316 arg1 changes 251:257 specific changes 242:257 specific changes in glycosylation 242:274 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 0 34 theme glycan 139:144 arg1 discovery 156:164 glycan biomarker discovery 139:164 glycan biomarker discovery 139:164 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 7 35 theme elevated 1320:1327 arg1 branching 1329:1337 elevated branching 1320:1337 elevated branching as well as increased antennary fucosylation 1320:1381 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 3 36 theme abundant 596:603 arg1 ones 605:608 the most abundant ones 587:608 the most abundant ones 587:608 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 3 36 theme abundant 596:603 arg1 structures 575:584 12 glycan structures 565:584 12 glycan structures 565:584 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 7 37 theme increased 1350:1358 arg1 fucosylation 1370:1381 increased antennary fucosylation 1350:1381 elevated branching as well as increased antennary fucosylation 1320:1381 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 1 38 theme many 218:221 arg1 diseases 229:236 many human diseases 218:236 many human diseases 218:236 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 0 39 theme 34 18:19 arg1 isomers 30:36 34 N-glycan isomers 18:36 34 N-glycan isomers in human serum 18:51 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 6 40 theme cancer 1173:1178 arg1 EOC 1190:1192 EOC 1190:1192 EOC 1190:1192 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 6 40 theme cancer 1173:1178 arg1 patients 1180:1187 primary epithelial ovarian cancer patients 1146:1187 primary epithelial ovarian cancer patients (EOC) 1146:1193 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 6 41 theme epithelial 1154:1163 arg1 EOC 1190:1192 EOC 1190:1192 EOC 1190:1192 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 6 41 theme epithelial 1154:1163 arg1 patients 1180:1187 primary epithelial ovarian cancer patients 1146:1187 primary epithelial ovarian cancer patients (EOC) 1146:1193 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 3 42 theme serum 687:691 arg1 modulations 666:676 glycome modulations 658:676 glycome modulations of total serum in the course of the diseases 658:721 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 2 43 theme Capillary 331:339 arg1 electrophoresis 341:355 Capillary electrophoresis 331:355 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) 331:404 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 2 43 theme Capillary 331:339 arg1 method 418:423 a robust method 409:423 a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis 409:535 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 1 44 theme human 223:227 arg1 diseases 229:236 many human diseases 218:236 many human diseases 218:236 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 6 45 theme glycan 1115:1120 arg1 isomers 1122:1128 34 glycan isomers 1112:1128 34 glycan isomers 1112:1128 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 9 46 theme biomarker 1701:1709 arg1 discovery 1711:1719 glycan biomarker discovery 1694:1719 glycan biomarker discovery 1694:1719 The results of the current study provide an improved dataset to be used in glycan biomarker discovery. 25234305 4 47 theme Lewis 838:842 arg1 epitope 847:853 a Lewis(x) epitope 836:853 a Lewis(x) epitope 836:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 0 48 theme isomers 30:36 arg1 Identification 0:13 Identification 0:13 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence 0:120 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 9 49 theme glycan 1694:1699 arg1 discovery 1711:1719 glycan biomarker discovery 1694:1719 glycan biomarker discovery 1694:1719 The results of the current study provide an improved dataset to be used in glycan biomarker discovery. 25234305 0 50 theme biomarker 146:154 arg1 discovery 156:164 glycan biomarker discovery 139:164 glycan biomarker discovery 139:164 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 2 51 theme 8-aminopyrene-1,3,6-trisulfonic 482:512 arg1 acid 514:517 8-aminopyrene-1,3,6-trisulfonic acid 482:517 8-aminopyrene-1,3,6-trisulfonic acid 482:517 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 7 52 from significant 1278:1288 arg1 samples 1301:1307 the EOC samples 1293:1307 the EOC samples 1293:1307 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 1 53 located observed 206:213 arg2 Alterations 167:177 Alterations 167:177 Alterations in glycosylation 167:194 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 1 53 located observed 206:213 arg1 diseases 229:236 many human diseases 218:236 many human diseases 218:236 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 4 54 theme x 844:844 arg1 epitope 847:853 a Lewis(x) epitope 836:853 a Lewis(x) epitope 836:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 5 55 theme positional 899:908 arg1 isomers 917:923 positional glycan isomers 899:923 positional glycan isomers 899:923 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 0 56 theme human 41:45 arg1 serum 47:51 human serum 41:51 human serum 41:51 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 8 57 theme tetraantennary 1548:1561 arg1 N-glycans 1563:1571 core-fucosylated tetraantennary N-glycans 1531:1571 tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1496:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 8 58 theme first 1406:1410 arg1 time 1412:1415 the first time 1402:1415 the first time 1402:1415 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 9 59 theme improved 1663:1670 arg1 dataset 1672:1678 an improved dataset 1660:1678 an improved dataset to be used in glycan biomarker discovery 1660:1719 The results of the current study provide an improved dataset to be used in glycan biomarker discovery. 25234305 5 60 theme glycan 910:915 arg1 isomers 917:923 positional glycan isomers 899:923 positional glycan isomers 899:923 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 6 61 theme patients 1180:1187 arg1 serum 1137:1141 the serum 1133:1141 the serum of primary epithelial ovarian cancer patients (EOC) 1133:1193 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 4 62 theme triantennary 803:814 arg1 structures 816:825 triantennary structures 803:825 triantennary structures carrying a Lewis(x) epitope 803:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 3 63 theme glycome 658:664 arg1 modulations 666:676 glycome modulations 658:676 glycome modulations of total serum in the course of the diseases 658:721 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 7 64 theme antennary 1360:1368 arg1 fucosylation 1370:1381 increased antennary fucosylation 1350:1381 elevated branching as well as increased antennary fucosylation 1320:1381 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 6 65 theme 34 1112:1113 arg1 isomers 1122:1128 34 glycan isomers 1112:1128 34 glycan isomers 1112:1128 It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). 25234305 1 66 theme specific 242:249 arg1 information 318:328 relevant diagnostic information 298:328 relevant diagnostic information 298:328 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 1 66 theme specific 242:249 arg1 changes 251:257 specific changes 242:257 specific changes in glycosylation 242:274 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 1 67 from changes 251:257 arg1 glycosylation 262:274 glycosylation 262:274 glycosylation 262:274 Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. 25234305 8 68 theme antennary 1453:1461 arg1 fucosylation 1463:1474 not only antennary fucosylation 1444:1474 not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans 1444:1571 For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. 25234305 5 69 theme standard 991:998 arg1 glycoproteins 1000:1012 standard glycoproteins 991:1012 standard glycoproteins 991:1012 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 7 70 from samples 1301:1307 arg1 significant 1278:1288 significant 1278:1288 significant 1278:1288 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 2 71 theme laser-induced 370:382 arg1 CE-LIF 398:403 CE-LIF 398:403 CE-LIF 398:403 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 2 71 theme laser-induced 370:382 arg1 fluorescence 384:395 laser-induced fluorescence 370:395 laser-induced fluorescence (CE-LIF) 370:404 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 4 72 contain carrying 827:834 arg1 structures 816:825 triantennary structures 803:825 triantennary structures carrying a Lewis(x) epitope 803:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 4 72 contain carrying 827:834 arg2 epitope 847:853 a Lewis(x) epitope 836:853 a Lewis(x) epitope 836:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 0 73 theme capillary 56:64 arg1 electrophoresis 66:80 capillary electrophoresis 56:80 capillary electrophoresis coupled with laser-induced fluorescence 56:120 Identification of 34 N-glycan isomers in human serum by capillary electrophoresis coupled with laser-induced fluorescence allows improving glycan biomarker discovery. 25234305 5 74 theme novel 1053:1057 arg1 structures 1059:1068 novel structures 1053:1068 novel structures 1053:1068 In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. 25234305 2 75 theme robust 411:416 arg1 electrophoresis 341:355 Capillary electrophoresis 331:355 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) 331:404 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 2 75 theme robust 411:416 arg1 method 418:423 a robust method 409:423 a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis 409:535 Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. 25234305 4 76 theme structures 816:825 arg1 increase 791:798 the increase 787:798 the increase of triantennary structures carrying a Lewis(x) epitope 787:853 In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewis(x) epitope. 25234305 3 77 theme glycan 568:573 arg1 ones 605:608 the most abundant ones 587:608 the most abundant ones 587:608 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 3 77 theme glycan 568:573 arg1 structures 575:584 12 glycan structures 565:584 12 glycan structures 565:584 To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. 25234305 7 78 theme diantennary 1214:1224 arg1 structures 1226:1235 diantennary structures 1214:1235 diantennary structures 1214:1235 Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. 25234305 9 79 theme current 1638:1644 arg1 study 1646:1650 the current study 1634:1650 the current study 1634:1650 The results of the current study provide an improved dataset to be used in glycan biomarker discovery. 29085933 4 0 theme glycan 1039:1044 arg1 retention 1046:1054 glycan retention 1039:1054 glycan retention on LC columns 1039:1068 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 4 1 theme favorable 724:732 arg1 characteristics 734:748 favorable characteristics 724:748 favorable characteristics 724:748 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 4 2 theme derivatization 915:928 arg1 methods 930:936 derivatization methods 915:936 derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 915:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 3 3 theme mass 536:539 arg1 spectrometry 541:552 liquid chromatography mass spectrometry 514:552 liquid chromatography mass spectrometry (LC-MS) 514:560 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 3 theme mass 536:539 arg1 LC-MS 555:559 LC-MS 555:559 LC-MS 555:559 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 4 with reduction 1297:1305 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 5 5 theme procainamide 1241:1252 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 6 6 theme highest 1425:1431 arg1 enhancement 1443:1453 the highest MS signal enhancement 1421:1453 the highest MS signal enhancement 1421:1453 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 5 7 with labeling 1287:1294 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 1 8 theme protein 181:187 arg1 folding 189:195 protein folding 181:195 protein folding 181:195 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 5 9 theme reduction 1297:1305 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 4 10 theme available 938:946 arg1 methods 930:936 derivatization methods 915:936 derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 915:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 6 11 theme sialylated 1565:1574 arg1 glycans 1576:1582 sialylated glycans 1565:1582 sialylated glycans 1565:1582 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 2 12 theme proteins 305:312 arg1 proteins 305:312 proteins 305:312 proteins 305:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 2 12 theme proteins 305:312 arg1 residues 293:300 asparagine residues 282:300 asparagine residues of proteins 282:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 4 13 theme LC-MS 699:703 arg1 analyses 705:712 LC-MS analyses 699:712 LC-MS analyses 699:712 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 5 14 theme aminoxyTMT 1255:1264 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 5 15 theme derivatizing 1135:1146 arg1 reagents 1148:1155 derivatizing reagents 1135:1155 derivatizing reagents 1135:1155 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 4 16 theme separation 804:813 arg1 efficiency 815:824 LC separation efficiency 801:824 LC separation efficiency 801:824 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 5 17 theme comprehensive 1195:1207 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 3 18 theme glycomic 580:587 arg1 studies 589:595 quantitative glycomic studies 567:595 quantitative glycomic studies 567:595 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 19 theme liquid 514:519 arg1 spectrometry 541:552 liquid chromatography mass spectrometry 514:552 liquid chromatography mass spectrometry (LC-MS) 514:560 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 19 theme liquid 514:519 arg1 LC-MS 555:559 LC-MS 555:559 LC-MS 555:559 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 20 theme structural 445:454 arg1 complexity 456:465 compositional and structural complexity 427:465 complexity 456:465 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 0 21 theme LC-MS/MS 51:58 arg1 analysis 60:67 LC-MS/MS analysis 51:67 LC-MS/MS analysis of N-glycans 51:80 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 29085933 5 22 with reduction 1311:1319 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 5 23 with 2-aminobenzamide 1223:1238 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 1 24 theme Protein 83:89 arg1 modification 136:147 a common post-translational modification 108:147 a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function 108:246 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 1 24 theme Protein 83:89 arg1 glycosylation 91:103 Protein glycosylation 83:103 Protein glycosylation 83:103 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 4 25 theme MS 1074:1075 arg1 analyses 1077:1084 MS analyses 1074:1084 MS analyses 1074:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 5 26 theme LC 1161:1162 arg1 columns 1164:1170 LC columns 1161:1170 LC columns 1161:1170 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 3 27 theme significant 599:609 arg1 challenge 611:619 a significant challenge 597:619 a significant challenge 597:619 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 28 theme reagents 1148:1155 arg1 selection 1122:1130 the proper selection 1111:1130 the proper selection of derivatizing reagents and LC columns 1111:1170 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 3 29 theme compositional 427:439 arg1 complexity 456:465 compositional and structural complexity 427:465 complexity 456:465 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 0 30 theme Direct 0:5 arg1 comparison 7:16 Direct comparison 0:16 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 0:81 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 29085933 4 31 dep improved 759:766 arg1 increased 791:799 increased 791:799 increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 791:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 4 32 dep such 751:754 arg1 as 756:757 as 756:757 as 756:757 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 5 33 theme proper 1115:1120 arg1 selection 1122:1130 the proper selection 1111:1130 the proper selection of derivatizing reagents and LC columns 1111:1170 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 0 34 theme derivatization 21:34 arg1 strategies 36:45 derivatization strategies 21:45 derivatization strategies 21:45 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 29085933 4 35 from analyses 1077:1084 arg1 columns 1062:1068 LC columns 1059:1068 LC columns 1059:1068 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 6 36 theme MS 1433:1434 arg1 enhancement 1443:1453 the highest MS signal enhancement 1421:1453 the highest MS signal enhancement 1421:1453 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 4 37 theme LC 801:802 arg1 efficiency 815:824 LC separation efficiency 801:824 LC separation efficiency 801:824 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 6 38 theme MS 1524:1525 arg1 intensity 1527:1535 MS intensity 1524:1535 MS intensity 1524:1535 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 3 39 theme poor 480:483 arg1 efficiency 496:505 poor ionization efficiency 480:505 poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) 480:560 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 40 theme 2-aminobenzamide 1223:1238 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 4 41 theme glycans 970:976 arg1 analysis 958:965 LC-MS analysis 952:965 LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 952:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 1 42 theme common 110:115 arg1 modification 136:147 a common post-translational modification 108:147 a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function 108:246 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 1 42 theme common 110:115 arg1 glycosylation 91:103 Protein glycosylation 83:103 Protein glycosylation 83:103 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 3 43 theme ionization 485:494 arg1 efficiency 496:505 poor ionization efficiency 480:505 poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) 480:560 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 44 theme release 418:424 arg1 ease 401:404 the ease 397:404 the ease of N-glycan release 397:424 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 45 theme N-glycan 1345:1352 arg1 analysis 1354:1361 N-glycan analysis 1345:1361 N-glycan analysis 1345:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 6 46 theme derivatization 1371:1384 arg1 strategies 1386:1395 the derivatization strategies 1367:1395 the derivatization strategies examined 1367:1404 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 5 47 theme RapiFluor-MS 1267:1278 arg1 labeling 1287:1294 RapiFluor-MS (RFMS) labeling 1267:1294 RapiFluor-MS (RFMS) labeling 1267:1294 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 6 48 theme glycans 1576:1582 arg1 intensity 1527:1535 MS intensity 1524:1535 MS intensity 1524:1535 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 6 48 theme glycans 1576:1582 arg1 stability 1552:1560 structural stability 1541:1560 structural stability 1541:1560 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 0 49 theme strategies 36:45 arg1 comparison 7:16 Direct comparison 0:16 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 0:81 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 29085933 4 50 theme LC 1059:1060 arg1 columns 1062:1068 LC columns 1059:1068 LC columns 1059:1068 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 4 51 theme fragments 865:873 arg1 efficiency 815:824 LC separation efficiency 801:824 LC separation efficiency 801:824 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 4 51 theme fragments 865:873 arg1 production 834:843 the production 830:843 the production of more informative fragments 830:873 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 5 52 with aminoxyTMT 1255:1264 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 4 53 theme different 1001:1009 arg1 properties 1011:1020 different properties 1001:1020 different properties that affect both glycan retention on LC columns and MS analyses 1001:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 6 54 theme neutral 1459:1465 arg1 glycans 1467:1473 neutral glycans 1459:1473 neutral glycans 1459:1473 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 4 55 theme LC-MS 952:956 arg1 analysis 958:965 LC-MS analysis 952:965 LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 952:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 6 56 gly sialylated 1565:1574 arg1 glycans 1576:1582 sialylated glycans 1565:1582 sialylated glycans 1565:1582 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 1 57 theme post-translational 117:134 arg1 modification 136:147 a common post-translational modification 108:147 a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function 108:246 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 1 57 theme post-translational 117:134 arg1 glycosylation 91:103 Protein glycosylation 83:103 Protein glycosylation 83:103 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 3 58 theme N-glycan 409:416 arg1 release 418:424 N-glycan release 409:424 N-glycan release 409:424 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 59 theme columns 1164:1170 arg1 selection 1122:1130 the proper selection 1111:1130 the proper selection of derivatizing reagents and LC columns 1111:1170 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 5 60 with procainamide 1241:1252 arg1 permethylation 1326:1339 permethylation 1326:1339 permethylation 1326:1339 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 3 61 theme chromatography 521:534 arg1 spectrometry 541:552 liquid chromatography mass spectrometry 514:552 liquid chromatography mass spectrometry (LC-MS) 514:560 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 3 61 theme chromatography 521:534 arg1 LC-MS 555:559 LC-MS 555:559 LC-MS 555:559 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 5 62 theme reduction 1311:1319 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 4 63 from retention 1046:1054 arg1 columns 1062:1068 LC columns 1059:1068 LC columns 1059:1068 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 2 64 with compatibility 351:363 arg1 release 380:386 enzymatic release 370:386 enzymatic release 370:386 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 6 65 dep intensity 1527:1535 arg1 the 1520:1522 the 1520:1522 the 1520:1522 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 4 66 theme ionization 768:777 arg1 efficiency 779:788 ionization efficiency 768:788 ionization efficiency 768:788 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 2 67 attach attached 270:277 arg1 proteins 305:312 proteins 305:312 proteins 305:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 2 67 attach attached 270:277 arg1 residues 293:300 asparagine residues 282:300 asparagine residues of proteins 282:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 2 67 attach attached 270:277 arg2 N-Glycans 249:257 N-Glycans 249:257 N-Glycans 249:257 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 2 68 theme asparagine 282:291 arg1 proteins 305:312 proteins 305:312 proteins 305:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 2 68 theme asparagine 282:291 arg1 residues 293:300 asparagine residues 282:300 asparagine residues of proteins 282:312 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 3 69 theme quantitative 567:578 arg1 studies 589:595 quantitative glycomic studies 567:595 quantitative glycomic studies 567:595 Despite the ease of N-glycan release, compositional and structural complexity coupled with poor ionization efficiency during liquid chromatography mass spectrometry (LC-MS) make quantitative glycomic studies a significant challenge. 29085933 4 70 theme methods 930:936 arg1 number 905:910 a number 903:910 a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses 903:1084 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 0 71 theme N-glycans 72:80 arg1 analysis 60:67 LC-MS/MS analysis 51:67 LC-MS/MS analysis of N-glycans 51:80 Direct comparison of derivatization strategies for LC-MS/MS analysis of N-glycans. 29085933 4 72 theme informative 853:863 arg1 fragments 865:873 more informative fragments 848:873 more informative fragments 848:873 To overcome these challenges, glycans are almost always derivatized prior to LC-MS analyses to impart favorable characteristics, such as improved ionization efficiency, increased LC separation efficiency and the production of more informative fragments during tandem MS. There are a number of derivatization methods available for LC-MS analysis of glycans, each of which imparts different properties that affect both glycan retention on LC columns and MS analyses. 29085933 1 73 theme significant 158:168 arg1 impacts 170:176 significant impacts 158:176 significant impacts 158:176 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 6 74 theme structural 1541:1550 arg1 stability 1552:1560 structural stability 1541:1560 structural stability 1541:1560 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 5 75 theme RFMS 1281:1284 arg1 labeling 1287:1294 RapiFluor-MS (RFMS) labeling 1267:1294 RapiFluor-MS (RFMS) labeling 1267:1294 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 29085933 6 76 theme signal 1436:1441 arg1 enhancement 1443:1453 the highest MS signal enhancement 1421:1453 the highest MS signal enhancement 1421:1453 Of the derivatization strategies examined, RFMS provided the highest MS signal enhancement for neutral glycans, while permethylation significantly enhanced the MS intensity and structural stability of sialylated glycans. 29085933 1 77 contain has 154:156 arg2 impacts 170:176 significant impacts 158:176 significant impacts 158:176 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 1 77 contain has 154:156 arg1 modification 136:147 a common post-translational modification 108:147 a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function 108:246 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 1 77 contain has 154:156 arg1 glycosylation 91:103 Protein glycosylation 83:103 Protein glycosylation 83:103 Protein glycosylation is a common post-translational modification that has significant impacts on protein folding, lifespan, conformation, distribution and function. 29085933 2 78 theme enzymatic 370:378 arg1 release 380:386 enzymatic release 370:386 enzymatic release 370:386 N-Glycans, which are attached to asparagine residues of proteins, are studied most often due to their compatibility with enzymatic release. 29085933 5 79 theme labeling 1287:1294 arg1 assessment 1209:1218 a comprehensive assessment 1193:1218 a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis 1193:1361 To provide guidance for the proper selection of derivatizing reagents and LC columns, herein, we describe a comprehensive assessment of 2-aminobenzamide, procainamide, aminoxyTMT, RapiFluor-MS (RFMS) labeling, reduction and reduction with permethylation for N-glycan analysis. 27853858 2 0 theme embryonic 485:493 arg1 kidney 495:500 human embryonic kidney 293 479:504 human embryonic kidney 293 cells 479:510 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 0 1 from reproducibility 19:33 arg1 cells 106:110 HEK293 cells 99:110 HEK293 cells with anti-PD1 antibody 99:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 2 2 theme antibody 604:611 arg1 yield 613:617 antibody yield 604:617 antibody yield 604:617 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 4 3 from batches 835:841 arg1 antibody 811:818 antibody 811:818 antibody from different batches of production 811:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 4 3 from batches 835:841 arg1 forms 802:806 major glycan forms 789:806 major glycan forms of antibody from different batches of production 789:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 5 4 theme Glycan 913:918 arg1 compositions 920:931 Glycan compositions 913:931 Glycan compositions of the antibody harvested at different time periods 913:983 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 2 5 from viability 571:579 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 0 6 theme expression 85:94 arg1 reproducibility 19:33 reproducibility 19:33 reproducibility 19:33 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 6 theme expression 85:94 arg1 consistency 55:65 product quality consistency 39:65 product quality consistency 39:65 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 6 theme expression 85:94 arg1 protein 148:154 the model protein 138:154 the model protein 138:154 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 3 7 theme binding 693:699 arg1 affinity 701:708 binding affinity 693:708 binding affinity 693:708 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 1 8 theme biopharmaceutical 327:343 arg1 development 345:355 biopharmaceutical development 327:355 biopharmaceutical development 327:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 1 9 theme crucial 256:262 arg1 issues 264:269 the most crucial issues 247:269 the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development 247:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 10 from batches 440:446 arg1 cells 506:510 human embryonic kidney 293 cells 479:510 human embryonic kidney 293 cells 479:510 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 4 11 theme glycosylation 886:898 arg1 consistency 900:910 glycosylation consistency 886:910 glycosylation consistency 886:910 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 7 12 theme TGE 1390:1392 arg1 batches 1394:1400 different TGE batches 1380:1400 different TGE batches 1380:1400 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 6 13 theme product 1190:1196 arg1 yield 1198:1202 product yield 1190:1202 product yield 1190:1202 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 1 14 theme issues 264:269 arg1 issues 264:269 the most crucial issues 247:269 the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development 247:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 1 14 theme issues 264:269 arg1 one 240:242 one 240:242 one 240:242 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 0 15 theme HEK293 99:104 arg1 cells 106:110 HEK293 cells 99:110 HEK293 cells with anti-PD1 antibody 99:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 6 16 theme secondary 1274:1282 arg1 structure 1284:1292 secondary structure 1274:1292 secondary structure 1274:1292 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 17 theme different 1111:1119 arg1 batches 1125:1131 different TGE batches 1111:1131 different TGE batches 1111:1131 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 7 18 theme culture 1428:1434 arg1 time 1436:1439 cell culture time 1423:1439 cell culture time 1423:1439 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 0 19 theme anti-PD1 117:124 arg1 antibody 126:133 anti-PD1 antibody 117:133 anti-PD1 antibody 117:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 20 from consistency 55:65 arg1 cells 106:110 HEK293 cells 99:110 HEK293 cells with anti-PD1 antibody 99:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 2 21 theme culture 627:633 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 6 22 theme growth 1182:1187 arg1 affinity 1264:1271 binding affinity 1256:1271 binding affinity 1256:1271 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 22 theme growth 1182:1187 arg1 structure 1284:1292 secondary structure 1274:1292 secondary structure 1274:1292 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 22 theme growth 1182:1187 arg1 stability 1307:1315 thermal stability 1299:1315 thermal stability 1299:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 22 theme growth 1182:1187 arg1 qualities 1217:1225 overall cell growth, product yield, and product qualities 1169:1225 overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability 1169:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 22 theme growth 1182:1187 arg1 point 1249:1253 isoelectric point 1237:1253 isoelectric point 1237:1253 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 1 23 theme reproducibility 174:188 arg1 Demonstration 157:169 Demonstration 157:169 Demonstration of reproducibility and consistency of process and product quality 157:235 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 24 theme cell 622:625 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 6 25 theme cell 1177:1180 arg1 growth 1182:1187 overall cell growth 1169:1187 overall cell growth 1169:1187 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 2 26 from status 592:597 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 2 27 theme human 479:483 arg1 kidney 495:500 human embryonic kidney 293 479:504 human embryonic kidney 293 cells 479:510 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 6 28 theme binding 1256:1262 arg1 affinity 1264:1271 binding affinity 1256:1271 binding affinity 1256:1271 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 29 theme overall 1169:1175 arg1 growth 1182:1187 overall cell growth 1169:1187 overall cell growth 1169:1187 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 3 30 theme thermal 736:742 arg1 stability 744:752 thermal stability 736:752 thermal stability 736:752 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 1 31 theme consistency 194:204 arg1 Demonstration 157:169 Demonstration 157:169 Demonstration of reproducibility and consistency of process and product quality 157:235 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 32 theme viable 550:555 arg1 density 562:568 viable cell density 550:568 viable cell density 550:568 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 4 33 theme production 846:855 arg1 batches 835:841 different batches 825:841 different batches of production 825:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 0 34 with cells 106:110 arg1 antibody 126:133 anti-PD1 antibody 117:133 anti-PD1 antibody 117:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 5 35 theme different 962:970 arg1 periods 977:983 different time periods 962:983 different time periods 962:983 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 7 36 theme different 1380:1388 arg1 batches 1394:1400 different TGE batches 1380:1400 different TGE batches 1380:1400 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 7 37 theme major 1331:1335 arg1 compositions 1346:1357 major N-glycan compositions 1331:1357 major N-glycan compositions 1331:1357 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 1 38 theme transient 280:288 arg1 technology 312:321 transient gene expression (TGE) technology 280:321 transient gene expression (TGE) technology for biopharmaceutical development 280:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 5 39 theme time 972:975 arg1 periods 977:983 different time periods 962:983 different time periods 962:983 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 1 40 theme process 209:215 arg1 consistency 194:204 consistency 194:204 consistency 194:204 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 1 40 theme process 209:215 arg1 reproducibility 174:188 reproducibility 174:188 reproducibility 174:188 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 1 41 theme gene 290:293 arg1 technology 312:321 transient gene expression (TGE) technology 280:321 transient gene expression (TGE) technology for biopharmaceutical development 280:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 42 theme cell 557:560 arg1 density 562:568 viable cell density 550:568 viable cell density 550:568 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 3 43 theme Product 648:654 arg1 quality 656:662 Product quality 648:662 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability 648:752 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 3 43 theme Product 648:654 arg1 well 770:773 well 770:773 well 770:773 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 7 44 theme cell 1423:1426 arg1 time 1436:1439 cell culture time 1423:1439 cell culture time 1423:1439 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 2 45 theme TGE 417:419 arg1 consistency 402:412 the production consistency 387:412 the production consistency of TGE 387:419 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 0 46 theme quality 47:53 arg1 reproducibility 19:33 reproducibility 19:33 reproducibility 19:33 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 46 theme quality 47:53 arg1 consistency 55:65 product quality consistency 39:65 product quality consistency 39:65 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 46 theme quality 47:53 arg1 protein 148:154 the model protein 138:154 the model protein 138:154 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 1 47 theme expression 295:304 arg1 technology 312:321 transient gene expression (TGE) technology 280:321 transient gene expression (TGE) technology for biopharmaceutical development 280:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 0 48 theme model 142:146 arg1 reproducibility 19:33 reproducibility 19:33 reproducibility 19:33 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 48 theme model 142:146 arg1 consistency 55:65 product quality consistency 39:65 product quality consistency 39:65 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 48 theme model 142:146 arg1 protein 148:154 the model protein 138:154 the model protein 138:154 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 49 from expression 85:94 arg1 cells 106:110 HEK293 cells 99:110 HEK293 cells with anti-PD1 antibody 99:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 4 50 theme antibody 811:818 arg1 forms 802:806 major glycan forms 789:806 major glycan forms of antibody from different batches of production 789:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 1 51 theme product 221:227 arg1 quality 229:235 product quality 221:235 product quality 221:235 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 0 52 theme product 39:45 arg1 reproducibility 19:33 reproducibility 19:33 reproducibility 19:33 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 52 theme product 39:45 arg1 consistency 55:65 product quality consistency 39:65 product quality consistency 39:65 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 52 theme product 39:45 arg1 protein 148:154 the model protein 138:154 the model protein 138:154 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 2 53 theme antibody 467:474 arg1 batches 440:446 nine batches 435:446 nine batches of recombinant IgG antibody in human embryonic kidney 293 cells 435:510 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 7 54 theme N-glycan 1337:1344 arg1 compositions 1346:1357 major N-glycan compositions 1331:1357 major N-glycan compositions 1331:1357 Furthermore, major N-glycan compositions are consistent among different TGE batches and conserved during cell culture time. 27853858 5 55 theme culture 1049:1055 arg1 time 1057:1060 the culture time 1045:1060 the culture time 1045:1060 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 6 56 from reproducible 1137:1148 arg1 affinity 1264:1271 binding affinity 1256:1271 binding affinity 1256:1271 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 56 from reproducible 1137:1148 arg1 structure 1284:1292 secondary structure 1274:1292 secondary structure 1274:1292 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 56 from reproducible 1137:1148 arg1 stability 1307:1315 thermal stability 1299:1315 thermal stability 1299:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 56 from reproducible 1137:1148 arg1 qualities 1217:1225 overall cell growth, product yield, and product qualities 1169:1225 overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability 1169:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 56 from reproducible 1137:1148 arg1 point 1249:1253 isoelectric point 1237:1253 isoelectric point 1237:1253 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 1 57 theme quality 229:235 arg1 consistency 194:204 consistency 194:204 consistency 194:204 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 1 57 theme quality 229:235 arg1 reproducibility 174:188 reproducibility 174:188 reproducibility 174:188 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 58 theme IgG 463:465 arg1 antibody 467:474 recombinant IgG antibody 451:474 recombinant IgG antibody 451:474 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 1 59 theme TGE 307:309 arg1 technology 312:321 transient gene expression (TGE) technology 280:321 transient gene expression (TGE) technology for biopharmaceutical development 280:355 Demonstration of reproducibility and consistency of process and product quality is one of the most crucial issues in using transient gene expression (TGE) technology for biopharmaceutical development. 27853858 2 60 theme apoptotic 582:590 arg1 status 592:597 apoptotic status 582:597 apoptotic status 582:597 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 2 61 theme kidney 495:500 arg1 cells 506:510 human embryonic kidney 293 cells 479:510 human embryonic kidney 293 cells 479:510 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 4 62 theme different 825:833 arg1 batches 835:841 different batches 825:841 different batches of production 825:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 2 63 theme recombinant 451:461 arg1 antibody 467:474 recombinant IgG antibody 451:474 recombinant IgG antibody 451:474 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 2 64 theme production 391:400 arg1 consistency 402:412 the production consistency 387:412 the production consistency of TGE 387:419 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 6 65 from lot 1155:1157 arg1 reproducible 1137:1148 reproducible 1137:1148 reproducible 1137:1148 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 66 theme yield 1198:1202 arg1 affinity 1264:1271 binding affinity 1256:1271 binding affinity 1256:1271 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 66 theme yield 1198:1202 arg1 structure 1284:1292 secondary structure 1274:1292 secondary structure 1274:1292 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 66 theme yield 1198:1202 arg1 stability 1307:1315 thermal stability 1299:1315 thermal stability 1299:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 66 theme yield 1198:1202 arg1 qualities 1217:1225 overall cell growth, product yield, and product qualities 1169:1225 overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability 1169:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 66 theme yield 1198:1202 arg1 point 1249:1253 isoelectric point 1237:1253 isoelectric point 1237:1253 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 67 theme product 1209:1215 arg1 affinity 1264:1271 binding affinity 1256:1271 binding affinity 1256:1271 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 67 theme product 1209:1215 arg1 structure 1284:1292 secondary structure 1274:1292 secondary structure 1274:1292 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 67 theme product 1209:1215 arg1 stability 1307:1315 thermal stability 1299:1315 thermal stability 1299:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 67 theme product 1209:1215 arg1 qualities 1217:1225 overall cell growth, product yield, and product qualities 1169:1225 overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability 1169:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 67 theme product 1209:1215 arg1 point 1249:1253 isoelectric point 1237:1253 isoelectric point 1237:1253 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 4 68 theme major 789:793 arg1 forms 802:806 major glycan forms 789:806 major glycan forms of antibody from different batches of production 789:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 6 69 theme isoelectric 1237:1247 arg1 point 1249:1253 isoelectric point 1237:1253 isoelectric point 1237:1253 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 5 70 theme antibody 940:947 arg1 compositions 920:931 Glycan compositions 913:931 Glycan compositions of the antibody harvested at different time periods 913:983 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 6 71 from qualities 1217:1225 arg1 reproducible 1137:1148 reproducible 1137:1148 reproducible 1137:1148 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 6 72 theme thermal 1299:1305 arg1 stability 1307:1315 thermal stability 1299:1315 thermal stability 1299:1315 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 0 73 from cells 106:110 arg1 reproducibility 19:33 reproducibility 19:33 reproducibility 19:33 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 73 from cells 106:110 arg1 consistency 55:65 product quality consistency 39:65 product quality consistency 39:65 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 0 73 from cells 106:110 arg1 protein 148:154 the model protein 138:154 the model protein 138:154 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 5 74 theme N-glycan 1018:1025 arg1 distribution 1027:1038 N-glycan distribution 1018:1038 N-glycan distribution 1018:1038 Glycan compositions of the antibody harvested at different time periods were also measured to illustrate N-glycan distribution over the culture time. 27853858 4 75 from forms 802:806 arg1 batches 835:841 different batches 825:841 different batches of production 825:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 0 76 theme gene 80:83 arg1 expression 85:94 transient gene expression 70:94 transient gene expression in HEK293 cells with anti-PD1 antibody 70:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 3 77 theme isoelectric 674:684 arg1 point 686:690 isoelectric point 674:690 isoelectric point 674:690 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 2 78 from density 562:568 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 27853858 0 79 theme transient 70:78 arg1 expression 85:94 transient gene expression 70:94 transient gene expression in HEK293 cells with anti-PD1 antibody 70:133 Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. 27853858 4 80 theme glycan 795:800 arg1 forms 802:806 major glycan forms 789:806 major glycan forms of antibody from different batches of production 789:855 In addition, major glycan forms of antibody from different batches of production were compared to demonstrate glycosylation consistency. 27853858 3 81 theme secondary 711:719 arg1 structure 721:729 secondary structure 711:729 secondary structure 711:729 Product quality including isoelectric point, binding affinity, secondary structure, and thermal stability was assessed as well. 27853858 6 82 theme TGE 1121:1123 arg1 batches 1125:1131 different TGE batches 1111:1131 different TGE batches 1111:1131 From the results, it has been demonstrated that different TGE batches are reproducible from lot to lot in overall cell growth, product yield, and product qualities including isoelectric point, binding affinity, secondary structure, and thermal stability. 27853858 2 83 from yield 613:617 arg1 supernatant 635:645 cell culture supernatant 622:645 cell culture supernatant 622:645 In this study, we challenged the production consistency of TGE by expressing nine batches of recombinant IgG antibody in human embryonic kidney 293 cells to evaluate reproducibility including viable cell density, viability, apoptotic status, and antibody yield in cell culture supernatant. 26114302 0 0 theme Agrocybe 92:99 arg1 aegerita 101:108 an Agrocybe aegerita 89:108 an Agrocybe aegerita Lectin 89:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 1 1 theme reversible 176:185 arg1 modification 206:217 a reversible post-translational modification 174:217 a reversible post-translational modification that plays essential roles in many cellular pathways 174:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 1 theme reversible 176:185 arg1 N-acetylglucosaminylation 127:151 O-linked N-acetylglucosaminylation 118:151 O-linked N-acetylglucosaminylation (O-GlcNAcylation) 118:169 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 4 2 with structures 708:717 arg1 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 4 2 with structures 708:717 arg1 GlcNAc 750:755 GlcNAc 750:755 GlcNAc 750:755 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 1 3 theme post-translational 187:204 arg1 modification 206:217 a reversible post-translational modification 174:217 a reversible post-translational modification that plays essential roles in many cellular pathways 174:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 3 theme post-translational 187:204 arg1 N-acetylglucosaminylation 127:151 O-linked N-acetylglucosaminylation 118:151 O-linked N-acetylglucosaminylation (O-GlcNAcylation) 118:169 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 5 4 theme O-GlcNAcylated 1013:1026 arg1 proteins 1028:1035 O-GlcNAcylated proteins 1013:1035 O-GlcNAcylated proteins 1013:1035 Study on AAL2 may enable us to design a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently. 26114302 5 5 theme protein 959:965 arg1 probe 967:971 a protein probe 957:971 a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently 957:1052 Study on AAL2 may enable us to design a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently. 26114302 3 6 dep lectin 534:539 arg1 aegerita 525:532 Agrocybe aegerita 516:532 Agrocybe aegerita 516:532 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 0 7 theme aegerita 101:108 arg1 Lectin 110:115 an Agrocybe aegerita Lectin 89:115 an Agrocybe aegerita Lectin 89:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 3 8 theme AAL2 547:550 arg1 identification 446:459 the identification 442:459 the identification 442:459 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 5 9 used used 985:988 arg2 probe 967:971 a protein probe 957:971 a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently 957:1052 Study on AAL2 may enable us to design a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently. 26114302 4 10 theme essential 860:868 arg1 residues 851:858 residues 851:858 residues essential for the binding of terminal N-acetylglucosamine 851:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 3 11 from aegerita 500:507 arg1 AAL2 547:550 AAL2 547:550 AAL2 547:550 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 11 from aegerita 500:507 arg1 identification 446:459 the identification 442:459 the identification 442:459 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 11 from aegerita 500:507 arg1 lectin 466:471 a lectin 464:471 a lectin from the mushroom Agrocybe aegerita 464:507 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 11 from aegerita 500:507 arg1 lectin 534:539 i.e., Agrocybe aegerita lectin 2 510:541 lectin 534:539 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 2 12 theme suitable 333:340 arg1 probes 342:347 suitable probes 333:347 suitable probes 333:347 Research in this field, however, is hampered by the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins. 26114302 2 13 from Research 273:280 arg1 field 290:294 this field 285:294 this field 285:294 Research in this field, however, is hampered by the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins. 26114302 2 14 theme O-GlcNAcylated 389:402 arg1 proteins 404:411 the O-GlcNAcylated proteins 385:411 the O-GlcNAcylated proteins 385:411 Research in this field, however, is hampered by the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins. 26114302 0 15 theme Structural 0:9 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin 0:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 1 16 theme essential 230:238 arg1 roles 240:244 essential roles 230:244 essential roles 230:244 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 0 17 theme Specific 20:27 arg1 Recognition 29:39 Specific Recognition 20:39 Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin 20:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 3 18 theme used 658:661 arg1 probes 663:668 other currently used probes 642:668 other currently used probes 642:668 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 1 19 theme O-linked 118:125 arg1 O-GlcNAcylation 154:168 O-GlcNAcylation 154:168 O-GlcNAcylation 154:168 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 19 theme O-linked 118:125 arg1 N-acetylglucosaminylation 127:151 O-linked N-acetylglucosaminylation 118:151 O-linked N-acetylglucosaminylation (O-GlcNAcylation) 118:169 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 19 theme O-linked 118:125 arg1 modification 206:217 a reversible post-translational modification 174:217 a reversible post-translational modification that plays essential roles in many cellular pathways 174:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 3 20 theme mushroom 482:489 arg1 aegerita 500:507 the mushroom Agrocybe aegerita 478:507 the mushroom Agrocybe aegerita 478:507 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 4 21 theme crystal 700:706 arg1 structures 708:717 the crystal structures 696:717 the crystal structures of AAL2 696:725 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 3 22 theme lectin 466:471 arg1 identification 446:459 the identification 442:459 the identification 442:459 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 4 23 theme GlcNAc 820:825 arg1 recognition 827:837 GlcNAc recognition 820:837 GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine 820:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 1 24 theme many 249:252 arg1 pathways 263:270 many cellular pathways 249:270 many cellular pathways 249:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 0 25 theme Recognition 29:39 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin 0:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 3 26 theme other 642:646 arg1 probes 663:668 other currently used probes 642:668 other currently used probes 642:668 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 27 theme higher 603:608 arg1 affinities 610:619 higher affinities 603:619 higher affinities 603:619 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 1 28 theme cellular 254:261 arg1 pathways 263:270 many cellular pathways 249:270 many cellular pathways 249:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 0 29 theme Terminal 57:64 arg1 N-Acetylglucosamine 66:84 Non-Reducing Terminal N-Acetylglucosamine 44:84 Non-Reducing Terminal N-Acetylglucosamine 44:84 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 4 30 with complexes 735:743 arg1 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 GlcNAcβ1-3Galβ1-4GlcNAc 761:783 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 4 30 with complexes 735:743 arg1 GlcNAc 750:755 GlcNAc 750:755 GlcNAc 750:755 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 4 31 theme N-acetylglucosamine 898:916 arg1 binding 878:884 the binding 874:884 the binding of terminal N-acetylglucosamine 874:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 0 32 theme N-Acetylglucosamine 66:84 arg1 Recognition 29:39 Specific Recognition 20:39 Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin 20:115 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 3 33 theme terminal 569:576 arg1 N-acetylglucosamine 578:596 terminal N-acetylglucosamine 569:596 terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes 569:668 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 0 34 theme Non-Reducing 44:55 arg1 N-Acetylglucosamine 66:84 Non-Reducing Terminal N-Acetylglucosamine 44:84 Non-Reducing Terminal N-Acetylglucosamine 44:84 Structural Basis of Specific Recognition of Non-Reducing Terminal N-Acetylglucosamine by an Agrocybe aegerita Lectin. 26114302 4 35 theme terminal 889:896 arg1 N-acetylglucosamine 898:916 terminal N-acetylglucosamine 889:916 terminal N-acetylglucosamine 889:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 4 36 theme recognition 827:837 arg1 basis 811:815 the structural basis 796:815 the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine 796:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 3 37 theme Agrocybe 516:523 arg1 aegerita 525:532 Agrocybe aegerita 516:532 Agrocybe aegerita 516:532 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 38 theme i.e. 510:513 arg1 lectin 534:539 i.e., Agrocybe aegerita lectin 2 510:541 lectin 534:539 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 5 39 from Study 919:923 arg1 AAL2 928:931 AAL2 928:931 AAL2 928:931 Study on AAL2 may enable us to design a protein probe that can be used to identify and purify O-GlcNAcylated proteins more efficiently. 26114302 3 40 theme Agrocybe 491:498 arg1 aegerita 500:507 the mushroom Agrocybe aegerita 478:507 the mushroom Agrocybe aegerita 478:507 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 2 41 theme probes 342:347 arg1 lack 325:328 the lack 321:328 the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins 321:411 Research in this field, however, is hampered by the lack of suitable probes to identify, accumulate, and purify the O-GlcNAcylated proteins. 26114302 4 42 theme AAL2 722:725 arg1 complexes 735:743 its complexes 731:743 its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc 731:783 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 4 42 theme AAL2 722:725 arg1 structures 708:717 the crystal structures 696:717 the crystal structures of AAL2 696:725 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 3 43 from identification 446:459 arg1 aegerita 500:507 the mushroom Agrocybe aegerita 478:507 the mushroom Agrocybe aegerita 478:507 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 1 44 link O-linked 118:125 arg1 O-GlcNAcylation 154:168 O-GlcNAcylation 154:168 O-GlcNAcylation 154:168 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 44 link O-linked 118:125 arg1 N-acetylglucosaminylation 127:151 O-linked N-acetylglucosaminylation 118:151 O-linked N-acetylglucosaminylation (O-GlcNAcylation) 118:169 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 1 44 link O-linked 118:125 arg1 modification 206:217 a reversible post-translational modification 174:217 a reversible post-translational modification that plays essential roles in many cellular pathways 174:270 O-linked N-acetylglucosaminylation (O-GlcNAcylation) is a reversible post-translational modification that plays essential roles in many cellular pathways. 26114302 4 45 theme structural 800:809 arg1 basis 811:815 the structural basis 796:815 the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine 796:916 In this paper, we report the crystal structures of AAL2 and its complexes with GlcNAc and GlcNAcβ1-3Galβ1-4GlcNAc and reveal the structural basis of GlcNAc recognition by AAL2 and residues essential for the binding of terminal N-acetylglucosamine. 26114302 3 46 theme lectin 534:539 arg1 identification 446:459 the identification 442:459 the identification 442:459 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 47 with N-acetylglucosamine 578:596 arg1 specificity 625:635 specificity 625:635 specificity 625:635 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 26114302 3 47 with N-acetylglucosamine 578:596 arg1 affinities 610:619 higher affinities 603:619 higher affinities 603:619 We have previously reported the identification of a lectin from the mushroom Agrocybe aegerita, i.e., Agrocybe aegerita lectin 2, or AAL2, that could bind terminal N-acetylglucosamine with higher affinities and specificity than other currently used probes. 23942880 5 0 theme liquid 967:972 arg1 spectrometry 1001:1012 liquid chromatography-tandem mass spectrometry 967:1012 liquid chromatography-tandem mass spectrometry 967:1012 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 2 1 theme eukaryotic 406:415 arg1 cells 417:421 eukaryotic cells 406:421 eukaryotic cells 406:421 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 1 2 theme oligosaccharides 298:313 arg1 conversion 267:276 conversion 267:276 conversion of raffinose family oligosaccharides by levansucrase 267:329 α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. 23942880 2 3 theme α-Gal 534:538 arg1 reactions 521:529 transgalactosylation reactions 500:529 transgalactosylation reactions of α-Gal 500:538 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 2 4 from data 492:495 arg1 reactions 521:529 transgalactosylation reactions 500:529 transgalactosylation reactions of α-Gal 500:538 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 5 5 theme mass 996:999 arg1 spectrometry 1001:1012 liquid chromatography-tandem mass spectrometry 967:1012 liquid chromatography-tandem mass spectrometry 967:1012 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 1 6 theme transgalactosylation 203:222 arg1 reactions 224:232 transgalactosylation reactions 203:232 transgalactosylation reactions of α-galactosidase (α-Gal) 203:259 α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. 23942880 3 7 from reuteri 598:604 arg1 sequence 570:577 The α-Gal gene sequence 555:577 The α-Gal gene sequence from Lactobacillus reuteri 555:604 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 8 8 theme acceptor 1482:1489 arg1 sugars 1491:1496 alternative acceptor sugars 1470:1496 alternative acceptor sugars 1470:1496 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 0 9 theme Lactococcus 132:142 arg1 lactis 144:149 Lactococcus lactis 132:149 Lactococcus lactis 132:149 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 8 10 theme α-Gal 1391:1395 arg1 specificity 1376:1386 the structural specificity 1361:1386 the structural specificity of α-Gal 1361:1395 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 8 11 theme adhesion 1559:1566 arg1 prevention 1568:1577 α-GOS pathogen adhesion prevention 1544:1577 α-GOS pathogen adhesion prevention in mammalian hosts 1544:1596 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 4 12 theme galactosyl 835:844 arg1 acceptor 846:853 galactosyl acceptor 835:853 galactosyl acceptor 835:853 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 2 13 from receptors 393:401 arg1 cells 417:421 eukaryotic cells 406:421 eukaryotic cells 406:421 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 4 14 theme galactosyl 772:781 arg1 donor 783:787 galactosyl donor 772:787 galactosyl donor 772:787 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 0 15 from expression 71:80 arg1 reuteri 121:127 Lactobacillus reuteri 107:127 Lactobacillus reuteri 107:127 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 0 15 from expression 71:80 arg1 lactis 144:149 Lactococcus lactis 132:149 Lactococcus lactis 132:149 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 6 16 theme 1 → 6 1141:1145 arg1 -linkages 1147:1155 (1 → 6)-linkages 1140:1155 (1 → 6)-linkages 1140:1155 α-Gal of Lactobacillus reuteri formed (1 → 3)-, (1 → 4)- or (1 → 6)-linked α-GOS but exhibited a preference for formation of (1 → 6)-linkages. 23942880 6 17 link -linked 1082:1088 arg1 α-GOS 1090:1094 (1 → 6)-linked α-GOS 1075:1094 (1 → 6)-linked α-GOS 1075:1094 α-Gal of Lactobacillus reuteri formed (1 → 3)-, (1 → 4)- or (1 → 6)-linked α-GOS but exhibited a preference for formation of (1 → 6)-linkages. 23942880 7 18 link -linked 1313:1319 arg1 hetero-oligosaccharides 1321:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 7 19 theme reuteri 1249:1255 arg1 1 → 4 1295:1299 1 → 4 1295:1299 1 → 4 1295:1299 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 7 19 theme reuteri 1249:1255 arg1 α-Gal 1237:1241 α-Gal 1237:1241 α-Gal 1237:1241 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 2 20 theme molecular 434:442 arg1 decoys 444:449 molecular decoys 434:449 molecular decoys 434:449 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 5 21 theme most 886:889 arg1 types 899:903 most linkage types 886:903 most linkage types 886:903 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 5 22 dep composition 860:870 arg1 The 856:858 The 856:858 The 856:858 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 8 23 theme pathogen 1550:1557 arg1 prevention 1568:1577 α-GOS pathogen adhesion prevention 1544:1577 α-GOS pathogen adhesion prevention in mammalian hosts 1544:1596 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 1 24 theme α-galactosidase 237:251 arg1 reactions 224:232 transgalactosylation reactions 203:232 transgalactosylation reactions of α-galactosidase (α-Gal) 203:259 α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. 23942880 5 25 theme linkage 891:897 arg1 types 899:903 most linkage types 886:903 most linkage types 886:903 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 3 26 theme α-Gal 625:629 arg1 strain 640:645 an α-Gal negative strain 622:645 an α-Gal negative strain of Lactococcus lactis 622:667 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 7 27 theme -linked 1313:1319 arg1 hetero-oligosaccharides 1321:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 7 28 theme galactosyl 1212:1221 arg1 acceptors 1223:1231 suitable galactosyl acceptors 1203:1231 suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1203:1343 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 8 29 theme mammalian 1582:1590 arg1 hosts 1592:1596 mammalian hosts 1582:1596 mammalian hosts 1582:1596 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 5 30 theme α-GOS 908:912 arg1 sequence 873:880 sequence 873:880 sequence 873:880 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 5 30 theme α-GOS 908:912 arg1 composition 860:870 composition 860:870 composition 860:870 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 5 30 theme α-GOS 908:912 arg1 types 899:903 most linkage types 886:903 most linkage types 886:903 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 0 31 theme α-galacto-oligosaccharides 20:45 arg1 Characterization 0:15 Characterization 0:15 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 0:150 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 8 32 theme structural 1365:1374 arg1 specificity 1376:1386 the structural specificity 1361:1386 the structural specificity of α-Gal 1361:1395 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 0 33 from reuteri 121:127 arg1 α-galactosidases 85:100 α-galactosidases 85:100 α-galactosidases from Lactobacillus reuteri 85:127 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 0 33 from reuteri 121:127 arg1 expression 71:80 heterologous expression 58:80 heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis 58:149 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 3 34 theme α-Gal 559:563 arg1 sequence 570:577 The α-Gal gene sequence 555:577 The α-Gal gene sequence from Lactobacillus reuteri 555:604 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 2 35 theme bacterial 462:470 arg1 infection 472:480 bacterial infection 462:480 bacterial infection 462:480 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 0 36 theme heterologous 58:69 arg1 expression 71:80 heterologous expression 58:80 heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis 58:149 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 7 37 theme α-Gal 1237:1241 arg1 hetero-oligosaccharides 1321:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1237:1343 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 4 38 theme crude 721:725 arg1 extracts 732:739 crude cell extracts 721:739 crude cell extracts with melibiose or raffinose 721:767 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 7 39 theme suitable 1203:1210 arg1 acceptors 1223:1231 suitable galactosyl acceptors 1203:1231 suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides 1203:1343 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 8 40 from prevention 1568:1577 arg1 hosts 1592:1596 mammalian hosts 1582:1596 mammalian hosts 1582:1596 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 4 41 with extracts 732:739 arg1 raffinose 759:767 raffinose 759:767 raffinose 759:767 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 4 41 with extracts 732:739 arg1 melibiose 746:754 melibiose 746:754 melibiose 746:754 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 3 42 theme Lactococcus 650:660 arg1 lactis 662:667 Lactococcus lactis 650:667 Lactococcus lactis 650:667 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 5 43 theme acceptors 926:934 arg1 saccharides 936:946 acceptors saccharides 926:946 acceptors saccharides 926:946 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 2 44 theme transgalactosylation 500:519 arg1 reactions 521:529 transgalactosylation reactions 500:529 transgalactosylation reactions of α-Gal 500:538 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 2 45 contain have 358:361 arg2 potential 367:375 the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection 363:480 the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection 363:480 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 2 45 contain have 358:361 arg1 α-GOS 352:356 α-GOS 352:356 α-GOS 352:356 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 3 46 theme negative 631:638 arg1 strain 640:645 an α-Gal negative strain 622:645 an α-Gal negative strain of Lactococcus lactis 622:667 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 6 47 theme -linkages 1147:1155 arg1 formation 1127:1135 formation 1127:1135 formation of (1 → 6)-linkages 1127:1155 α-Gal of Lactobacillus reuteri formed (1 → 3)-, (1 → 4)- or (1 → 6)-linked α-GOS but exhibited a preference for formation of (1 → 6)-linkages. 23942880 7 48 theme 1 → 3 1285:1289 arg1 formation 1271:1279 formation 1271:1279 formation of (1 → 3)- 1271:1291 Fucose, N-acetylglucosamine and lactose were suitable galactosyl acceptors for α-Gal of L. reuteri, resulting in formation of (1 → 3)-, (1 → 4)- or (1 → 6)-linked hetero-oligosaccharides. 23942880 2 49 theme glycan 386:391 arg1 receptors 393:401 glycan receptors 386:401 glycan receptors on eukaryotic cells 386:421 Similarly to β-GOS, α-GOS have the potential to mimic glycan receptors on eukaryotic cells and act as molecular decoys to prevent bacterial infection; however, data on transgalactosylation reactions of α-Gal remain scarce. 23942880 8 50 theme alternative 1470:1480 arg1 sugars 1491:1496 alternative acceptor sugars 1470:1496 alternative acceptor sugars 1470:1496 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 6 51 theme reuteri 1038:1044 arg1 α-Gal 1015:1019 α-Gal 1015:1019 α-Gal of Lactobacillus reuteri 1015:1044 α-Gal of Lactobacillus reuteri formed (1 → 3)-, (1 → 4)- or (1 → 6)-linked α-GOS but exhibited a preference for formation of (1 → 6)-linkages. 23942880 6 52 theme -linked 1082:1088 arg1 α-GOS 1090:1094 (1 → 6)-linked α-GOS 1075:1094 (1 → 6)-linked α-GOS 1075:1094 α-Gal of Lactobacillus reuteri formed (1 → 3)-, (1 → 4)- or (1 → 6)-linked α-GOS but exhibited a preference for formation of (1 → 6)-linkages. 23942880 8 53 theme further 1518:1524 arg1 studies 1526:1532 further studies 1518:1532 further studies 1518:1532 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 8 54 theme oligosaccharides 1429:1444 arg1 variation 1416:1424 the variation 1412:1424 the variation of oligosaccharides produced by introducing alternative acceptor sugars 1412:1496 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 8 55 theme α-GOS 1544:1548 arg1 prevention 1568:1577 α-GOS pathogen adhesion prevention 1544:1577 α-GOS pathogen adhesion prevention in mammalian hosts 1544:1596 By determining the structural specificity of α-Gal and increasing the variation of oligosaccharides produced by introducing alternative acceptor sugars, this work supports further studies to assess α-GOS pathogen adhesion prevention in mammalian hosts. 23942880 3 56 theme lactis 662:667 arg1 strain 640:645 an α-Gal negative strain 622:645 an α-Gal negative strain of Lactococcus lactis 622:667 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 3 57 theme gene 565:568 arg1 sequence 570:577 The α-Gal gene sequence 555:577 The α-Gal gene sequence from Lactobacillus reuteri 555:604 The α-Gal gene sequence from Lactobacillus reuteri was cloned into an α-Gal negative strain of Lactococcus lactis. 23942880 4 58 theme cell 727:730 arg1 extracts 732:739 crude cell extracts 721:739 crude cell extracts with melibiose or raffinose 721:767 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 1 59 theme raffinose 281:289 arg1 oligosaccharides 298:313 raffinose family oligosaccharides 281:313 raffinose family oligosaccharides 281:313 α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. 23942880 0 60 theme α-galactosidases 85:100 arg1 expression 71:80 heterologous expression 58:80 heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis 58:149 Characterization of α-galacto-oligosaccharides formed via heterologous expression of α-galactosidases from Lactobacillus reuteri in Lactococcus lactis. 23942880 4 61 theme Transgalactosylation 670:689 arg1 reactions 691:699 Transgalactosylation reactions 670:699 Transgalactosylation reactions 670:699 Transgalactosylation reactions were achieved using crude cell extracts with melibiose or raffinose as galactosyl donor and fucose, N-acetylglucosamine or lactose as galactosyl acceptor. 23942880 5 62 theme chromatography-tandem 974:994 arg1 spectrometry 1001:1012 liquid chromatography-tandem mass spectrometry 967:1012 liquid chromatography-tandem mass spectrometry 967:1012 The composition, sequence and most linkage types of α-GOS formed with acceptors saccharides were determined by liquid chromatography-tandem mass spectrometry. 23942880 1 63 theme family 291:296 arg1 oligosaccharides 298:313 raffinose family oligosaccharides 281:313 raffinose family oligosaccharides 281:313 α-Galacto-oligosaccharides (α-GOS) are produced by transgalactosylation reactions of α-galactosidase (α-Gal) or by conversion of raffinose family oligosaccharides by levansucrase. 29233759 8 0 theme reduced 1121:1127 arg1 hydrolysis 1129:1138 reduced hydrolysis 1121:1138 reduced hydrolysis 1121:1138 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 6 1 theme increased 939:947 arg1 hydrolysis 949:958 increased hydrolysis 939:958 increased hydrolysis 939:958 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 9 2 theme Trp 1292:1294 arg1 residues 1296:1303 Trp residues 1292:1303 Trp residues 1292:1303 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 4 3 theme hydrolytic 670:679 arg1 activities 688:697 the hydrolytic and TG activities 666:697 activities 688:697 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 2 4 theme DxDxE 573:577 arg1 motif 579:583 the DxDxE motif 569:583 the DxDxE motif of StmChiA 569:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 0 5 theme A. 105:106 arg1 TG 128:129 TG 128:129 TG 128:129 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 0 5 theme A. 105:106 arg1 Transglycosylation 108:125 A. Transglycosylation 105:125 A. Transglycosylation (TG) by family 18 chitinases 105:154 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 6 6 theme TG 892:893 arg1 activity 895:902 Increased in the higher TG activity 868:902 Increased in the higher TG activity of mutant W306A 868:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 2 7 theme StmChiA 588:594 arg1 motif 579:583 the DxDxE motif 569:583 the DxDxE motif of StmChiA 569:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 8 8 theme TG 1194:1195 arg1 products 1197:1204 TG products 1194:1204 TG products 1194:1204 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 6 9 theme higher 885:890 arg1 activity 895:902 Increased in the higher TG activity 868:902 Increased in the higher TG activity of mutant W306A 868:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 0 10 theme biological 195:204 arg1 applications 206:217 the many biological applications 186:217 the many biological applications of long-chain chitooligosaccharides (CHOS) 186:260 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 4 11 from Changes 655:661 arg1 activities 688:697 the hydrolytic and TG activities 666:697 activities 688:697 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 1 12 theme current 270:276 arg1 study 278:282 the current study 266:282 the current study 266:282 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 2 13 theme -1 522:523 arg1 subsites 525:532 the -3 and -1 subsites 511:532 subsites 525:532 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 9 14 theme side 1277:1280 arg1 chains 1282:1287 the side chains 1273:1287 the side chains of Trp residues 1273:1303 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 0 15 theme many 190:193 arg1 applications 206:217 the many biological applications 186:217 the many biological applications of long-chain chitooligosaccharides (CHOS) 186:260 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 6 16 theme W306A 914:918 arg1 activity 895:902 Increased in the higher TG activity 868:902 Increased in the higher TG activity of mutant W306A 868:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 3 17 with latter 638:643 arg1 Asn 650:652 Asn 650:652 Asn 650:652 The former was replaced with Ala and the latter with Asn. 29233759 7 18 theme products 1025:1032 arg1 products 1025:1032 TG products 1022:1032 TG products 1022:1032 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 7 18 theme products 1025:1032 arg1 amounts 1011:1017 substantial amounts 999:1017 substantial amounts of TG products 999:1032 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 2 19 from subsites 525:532 arg1 residues 497:504 Trp residues 493:504 Trp residues from the -3 and -1 subsites 493:532 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 2 19 from subsites 525:532 arg1 Asp 560:562 the central catalytic Asp 538:562 the central catalytic Asp from the DxDxE motif of StmChiA 538:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 6 20 theme in 878:879 arg1 activity 895:902 Increased in the higher TG activity 868:902 Increased in the higher TG activity of mutant W306A 868:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 2 21 theme central 542:548 arg1 Asp 560:562 the central catalytic Asp 538:562 the central catalytic Asp from the DxDxE motif of StmChiA 538:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 10 22 theme long-chain 1546:1555 arg1 CHOS 1557:1560 long-chain CHOS 1546:1560 long-chain CHOS 1546:1560 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 4 23 theme product 746:752 arg1 profile 754:760 the product profile 742:760 the product profile of each mutant 742:775 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 10 24 theme mutant 1598:1603 arg1 D464N 1605:1609 the catalytic mutant D464N 1584:1609 the catalytic mutant D464N 1584:1609 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 2 25 theme -3 515:516 arg1 subsites 525:532 the -3 and -1 subsites 511:532 subsites 525:532 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 6 26 theme Increased 868:876 arg1 activity 895:902 Increased in the higher TG activity 868:902 Increased in the higher TG activity of mutant W306A 868:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 1 27 from activity 292:299 arg1 StmChiA 351:357 StmChiA 351:357 StmChiA 351:357 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 1 27 from activity 292:299 arg1 maltophilia 338:348 Stenotrophomonas maltophilia 321:348 Stenotrophomonas maltophilia (StmChiA) 321:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 2 28 theme independent 449:459 arg1 mutants 461:467 Three independent mutants 443:467 Three independent mutants 443:467 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 8 29 dep mutants 1090:1096 arg1 mutants 1090:1096 mutants 1090:1096 mutants D464N and W679A 1090:1112 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 8 29 dep mutants 1090:1096 arg1 W679A 1108:1112 W679A 1108:1112 W679A 1108:1112 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 8 29 dep mutants 1090:1096 arg1 D464N 1098:1102 D464N 1098:1102 D464N 1098:1102 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 5 30 theme increased 845:853 arg1 activity 858:865 increased TG activity 845:865 increased TG activity 845:865 All three mutants showed increased TG activity. 29233759 2 31 theme Trp 493:495 arg1 residues 497:504 Trp residues 493:504 Trp residues from the -3 and -1 subsites 493:532 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 0 32 theme chitooligosaccharides 233:253 arg1 applications 206:217 the many biological applications 186:217 the many biological applications of long-chain chitooligosaccharides (CHOS) 186:260 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 9 33 with interactions 1322:1333 arg1 residues 1346:1353 sugar residues 1340:1353 sugar residues 1340:1353 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 4 34 theme enzymes 706:712 arg1 activities 688:697 the hydrolytic and TG activities 666:697 activities 688:697 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 5 35 theme TG 855:856 arg1 activity 858:865 increased TG activity 845:865 increased TG activity 845:865 All three mutants showed increased TG activity. 29233759 0 36 theme Active-site 0:10 arg1 mutations 12:20 Active-site mutations 0:20 Active-site mutations 0:20 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 9 37 theme residues 1296:1303 arg1 chains 1282:1287 the side chains 1273:1287 the side chains of Trp residues 1273:1303 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 0 38 theme long-chain 222:231 arg1 CHOS 256:259 CHOS 256:259 CHOS 256:259 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 0 38 theme long-chain 222:231 arg1 chitooligosaccharides 233:253 long-chain chitooligosaccharides 222:253 long-chain chitooligosaccharides (CHOS) 222:260 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 8 39 dep 12h 1212:1214 arg1 up 1206:1207 up 1206:1207 up 1206:1207 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 9 40 theme residues 1419:1426 arg1 importance 1399:1408 the importance 1395:1408 the importance of these residues in the enzymatic activity of StmChiA 1395:1463 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 8 41 theme gradual 1170:1176 arg1 accumulation 1178:1189 the gradual accumulation 1166:1189 the gradual accumulation of TG products up to 12h 1166:1214 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 2 42 from motif 579:583 arg1 residues 497:504 Trp residues 493:504 Trp residues from the -3 and -1 subsites 493:532 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 2 42 from motif 579:583 arg1 Asp 560:562 the central catalytic Asp 538:562 the central catalytic Asp from the DxDxE motif of StmChiA 538:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 2 43 theme catalytic 550:558 arg1 Asp 560:562 the central catalytic Asp 538:562 the central catalytic Asp from the DxDxE motif of StmChiA 538:594 Three independent mutants were created, targeting Trp residues from the -3 and -1 subsites and the central catalytic Asp from the DxDxE motif of StmChiA. 29233759 10 44 dep mutants 1475:1481 arg1 W679A 1521:1525 W679A 1521:1525 W679A 1521:1525 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 10 44 dep mutants 1475:1481 arg1 W306A 1511:1515 W306A 1511:1515 W306A 1511:1515 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 10 44 dep mutants 1475:1481 arg1 mutants 1475:1481 mutants 1475:1481 mutants of the glycon-binding site (W306A and W679A) 1475:1526 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 4 45 theme high-performance 780:795 arg1 chromatography 804:817 high-performance liquid chromatography 780:817 high-performance liquid chromatography 780:817 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 1 46 theme TG 289:290 arg1 activity 292:299 the TG activity 285:299 the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) 285:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 0 47 theme family 135:140 arg1 chitinases 145:154 family 18 chitinases 135:154 family 18 chitinases 135:154 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 1 48 theme structure-guided 381:396 arg1 mutations 398:406 structure-guided mutations 381:406 structure-guided mutations within and around the active site 381:440 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 10 49 theme site 1505:1508 arg1 mutants 1475:1481 mutants 1475:1481 mutants of the glycon-binding site (W306A and W679A) 1475:1526 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 10 49 theme site 1505:1508 arg1 W306A 1511:1515 W306A 1511:1515 W306A 1511:1515 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 10 49 theme site 1505:1508 arg1 W679A 1521:1525 W679A 1521:1525 W679A 1521:1525 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 0 50 dep Stenotrophomonas 66:81 arg1 maltophilia 83:93 maltophilia 83:93 maltophilia 83:93 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 10 51 theme catalytic 1588:1596 arg1 D464N 1605:1609 the catalytic mutant D464N 1584:1609 the catalytic mutant D464N 1584:1609 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 9 52 theme enzymatic 1435:1443 arg1 activity 1445:1452 the enzymatic activity 1431:1452 the enzymatic activity of StmChiA 1431:1463 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 0 53 theme transglycosylation 35:52 arg1 activity 54:61 the transglycosylation activity 31:61 the transglycosylation activity of Stenotrophomonas maltophilia chitinase 31:103 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 7 54 theme first 1045:1049 arg1 15-30min 1051:1058 the first 15-30min 1041:1058 the first 15-30min of the reaction 1041:1074 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 7 55 theme substantial 999:1009 arg1 products 1025:1032 TG products 1022:1032 TG products 1022:1032 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 7 55 theme substantial 999:1009 arg1 amounts 1011:1017 substantial amounts 999:1017 substantial amounts of TG products 999:1032 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 0 56 theme Stenotrophomonas 66:81 arg1 chitinase 95:103 Stenotrophomonas maltophilia chitinase 66:103 Stenotrophomonas maltophilia chitinase 66:103 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 1 57 theme chitinase 304:312 arg1 A 314:314 chitinase A 304:314 chitinase A from Stenotrophomonas maltophilia (StmChiA) 304:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 0 58 theme special 162:168 arg1 interest 170:177 special interest 162:177 special interest 162:177 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 4 59 theme TG 685:686 arg1 activities 688:697 the hydrolytic and TG activities 666:697 activities 688:697 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 1 60 from maltophilia 338:348 arg1 activity 292:299 the TG activity 285:299 the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) 285:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 1 60 from maltophilia 338:348 arg1 A 314:314 chitinase A 304:314 chitinase A from Stenotrophomonas maltophilia (StmChiA) 304:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 1 61 theme A 314:314 arg1 activity 292:299 the TG activity 285:299 the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) 285:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 9 62 theme -3 1364:1365 arg1 subsites 1374:1381 the -3 and -1 subsites 1360:1381 subsites 1374:1381 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 9 63 theme Molecular 1217:1225 arg1 studies 1235:1241 Molecular docking studies 1217:1241 Molecular docking studies with chitohexaose 1217:1259 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 4 64 theme liquid 797:802 arg1 chromatography 804:817 high-performance liquid chromatography 780:817 high-performance liquid chromatography 780:817 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 9 65 theme StmChiA 1457:1463 arg1 activity 1445:1452 the enzymatic activity 1431:1452 the enzymatic activity of StmChiA 1431:1463 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 9 66 theme sugar 1340:1344 arg1 residues 1346:1353 sugar residues 1340:1353 sugar residues 1340:1353 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 6 67 theme mutant 907:912 arg1 W306A 914:918 mutant W306A 907:918 mutant W306A 907:918 Increased in the higher TG activity of mutant W306A was accompanied by increased hydrolysis. 29233759 9 68 theme docking 1227:1233 arg1 studies 1235:1241 Molecular docking studies 1217:1241 Molecular docking studies with chitohexaose 1217:1259 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 9 69 theme stacking 1313:1320 arg1 interactions 1322:1333 stacking interactions 1313:1333 stacking interactions with sugar residues 1313:1353 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 7 70 theme TG 1022:1023 arg1 products 1025:1032 TG products 1022:1032 TG products 1022:1032 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 1 71 theme active 430:435 arg1 site 437:440 the active site 426:440 the active site 426:440 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 7 72 theme reaction 1067:1074 arg1 15-30min 1051:1058 the first 15-30min 1041:1058 the first 15-30min of the reaction 1041:1074 However, this mutant also accumulated substantial amounts of TG products during the first 15-30min of the reaction. 29233759 0 73 theme chitinase 95:103 arg1 activity 54:61 the transglycosylation activity 31:61 the transglycosylation activity of Stenotrophomonas maltophilia chitinase 31:103 Active-site mutations improved the transglycosylation activity of Stenotrophomonas maltophilia chitinase A. Transglycosylation (TG) by family 18 chitinases is of special interest due to the many biological applications of long-chain chitooligosaccharides (CHOS). 29233759 4 74 theme mutant 770:775 arg1 profile 754:760 the product profile 742:760 the product profile of each mutant 742:775 Changes in the hydrolytic and TG activities of the enzymes were assessed by monitoring the product profile of each mutant by high-performance liquid chromatography. 29233759 10 75 theme glycon-binding 1490:1503 arg1 site 1505:1508 the glycon-binding site 1486:1508 the glycon-binding site 1486:1508 Overall, mutants of the glycon-binding site (W306A and W679A) appear to produce long-chain CHOS more efficiently than the catalytic mutant D464N. 29233759 8 76 theme products 1197:1204 arg1 accumulation 1178:1189 the gradual accumulation 1166:1189 the gradual accumulation of TG products up to 12h 1166:1214 In contrast, mutants D464N and W679A showed reduced hydrolysis, which was accompanied by the gradual accumulation of TG products up to 12h. 29233759 1 77 theme Stenotrophomonas 321:336 arg1 StmChiA 351:357 StmChiA 351:357 StmChiA 351:357 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 1 77 theme Stenotrophomonas 321:336 arg1 maltophilia 338:348 Stenotrophomonas maltophilia 321:348 Stenotrophomonas maltophilia (StmChiA) 321:358 In the current study, the TG activity of chitinase A from Stenotrophomonas maltophilia (StmChiA) was improved through structure-guided mutations within and around the active site. 29233759 9 78 from importance 1399:1408 arg1 activity 1445:1452 the enzymatic activity 1431:1452 the enzymatic activity of StmChiA 1431:1463 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 9 79 with studies 1235:1241 arg1 chitohexaose 1248:1259 chitohexaose 1248:1259 chitohexaose 1248:1259 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 29233759 9 80 theme -1 1371:1372 arg1 subsites 1374:1381 the -3 and -1 subsites 1360:1381 subsites 1374:1381 Molecular docking studies with chitohexaose showed that the side chains of Trp residues mediate stacking interactions with sugar residues from the -3 and -1 subsites, indicating the importance of these residues in the enzymatic activity of StmChiA. 26042789 4 0 theme fungus 595:600 arg1 PVL 624:626 PVL 624:626 PVL 624:626 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 4 0 theme fungus 595:600 arg1 velutina 614:621 the fungus Psathyrella velutina 591:621 the fungus Psathyrella velutina (PVL) 591:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 6 1 theme capped 950:955 arg1 branches 957:964 GlcNAcβ1-2Man capped branches 936:964 GlcNAcβ1-2Man capped branches 936:964 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 4 2 theme velutina 614:621 arg1 body 583:586 the fruiting body 570:586 the fruiting body of the fungus Psathyrella velutina (PVL) 570:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 6 3 theme plasmon 821:827 arg1 resonance 829:837 surface plasmon resonance 813:837 surface plasmon resonance 813:837 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 2 4 theme β-N-acetylglucosamine 303:323 arg1 residues 334:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues 281:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues 281:374 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 12 5 from agalacto-glycans 1689:1704 arg1 diseases 1725:1732 other diseases 1719:1732 other diseases 1719:1732 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 12 5 from agalacto-glycans 1689:1704 arg1 cancer 1709:1714 cancer 1709:1714 cancer 1709:1714 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 10 6 theme stomach 1451:1457 arg1 glands 1459:1464 stomach glands 1451:1464 stomach glands that present unique αGlcNAc-presenting mucins 1451:1510 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 2 7 theme terminal 281:288 arg1 residues 334:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues 281:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues 281:374 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 1 8 theme distinct 175:182 arg1 epitopes 200:207 distinct oligosaccharide epitopes 175:207 distinct oligosaccharide epitopes 175:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 6 9 theme biantennary 909:919 arg1 N-glycans 921:929 biantennary N-glycans 909:929 biantennary N-glycans with GlcNAcβ1-2Man capped branches 909:964 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 9 10 theme glycosyltransferase 1250:1268 arg1 expression 1270:1279 glycosyltransferase expression 1250:1279 glycosyltransferase expression 1250:1279 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 1 11 theme epitopes 200:207 arg1 alterations 119:129 alterations 119:129 alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 119:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 5 12 theme terminal 708:715 arg1 residues 724:731 terminal GlcNAc residues 708:731 terminal GlcNAc residues 708:731 A strong specificity for terminal GlcNAc residues was evidenced by glycan array. 26042789 2 13 theme GlcNAc 326:331 arg1 residues 334:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues 281:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues 281:374 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 4 14 theme good 650:653 arg1 yield 655:659 good yield 650:659 good yield in bacterial culture 650:680 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 9 15 theme higher 1295:1300 arg1 amount 1302:1307 the higher amount 1291:1307 the higher amount of GlcNAc present on cancer cells 1291:1341 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 9 15 theme higher 1295:1300 arg1 present 1319:1325 GlcNAc present 1312:1325 GlcNAc present 1312:1325 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 7 16 theme specificity 1061:1071 arg1 basis 1048:1052 the structural basis 1033:1052 the structural basis of the specificity 1033:1071 Crystal structure of PVL complexed with GlcNAcβ1-3Gal established the structural basis of the specificity. 26042789 9 17 theme present 1319:1325 arg1 amount 1302:1307 the higher amount 1291:1307 the higher amount of GlcNAc present on cancer cells 1291:1341 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 9 17 theme present 1319:1325 arg1 present 1319:1325 GlcNAc present 1312:1325 GlcNAc present 1312:1325 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 9 18 theme cancer 1330:1335 arg1 cells 1337:1341 cancer cells 1330:1341 cancer cells 1330:1341 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 7 19 theme structural 1037:1046 arg1 basis 1048:1052 the structural basis 1033:1052 the structural basis of the specificity 1033:1071 Crystal structure of PVL complexed with GlcNAcβ1-3Gal established the structural basis of the specificity. 26042789 12 20 theme other 1719:1723 arg1 diseases 1725:1732 other diseases 1719:1732 other diseases 1719:1732 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 10 21 theme unique 1479:1484 arg1 mucins 1505:1510 unique αGlcNAc-presenting mucins 1479:1510 unique αGlcNAc-presenting mucins 1479:1510 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 0 22 theme cancer 68:73 arg1 cells 75:79 human cancer cells 62:79 human cancer cells 62:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 8 23 theme several 1086:1092 arg1 types 1094:1098 several types 1086:1098 several types of cancer cells 1086:1114 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 11 24 theme clear 1553:1557 arg1 delineation 1559:1569 a clear delineation 1551:1569 a clear delineation 1551:1569 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 1 25 from structures 140:149 arg1 diseases 162:169 chronic diseases 154:169 chronic diseases 154:169 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 5 26 theme glycan 750:755 arg1 array 757:761 glycan array 750:761 glycan array 750:761 A strong specificity for terminal GlcNAc residues was evidenced by glycan array. 26042789 8 27 theme cells 1110:1114 arg1 types 1094:1098 several types 1086:1098 several types of cancer cells 1086:1114 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 7 28 theme PVL 988:990 arg1 structure 975:983 Crystal structure 967:983 Crystal structure of PVL complexed with GlcNAcβ1-3Gal 967:1019 Crystal structure of PVL complexed with GlcNAcβ1-3Gal established the structural basis of the specificity. 26042789 3 29 theme novel 443:447 arg1 markers 449:455 novel markers 443:455 novel markers that bind specifically to such epitopes 443:495 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 4 30 from body 583:586 arg1 lectin 558:563 A GlcNAc-binding lectin 541:563 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) 541:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 1 31 theme Cell 82:85 arg1 glycoconjugates 95:109 Cell surface glycoconjugates 82:109 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 82:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 12 32 theme useful 1664:1669 arg1 tool 1671:1674 a useful tool 1662:1674 a useful tool for labeling agalacto-glycans in cancer or other diseases 1662:1732 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 12 32 theme useful 1664:1669 arg1 PVL 1645:1647 PVL 1645:1647 PVL 1645:1647 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 3 33 theme unconventional 390:403 arg1 algi 430:433 algi 430:433 algi 430:433 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 3 33 theme unconventional 390:403 arg1 sources 405:411 unconventional sources 390:411 unconventional sources such as fungi or algi 390:433 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 3 33 theme unconventional 390:403 arg1 fungi 421:425 fungi 421:425 fungi 421:425 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 3 34 theme such 483:486 arg1 epitopes 488:495 such epitopes 483:495 such epitopes 483:495 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 0 35 theme fungal 14:19 arg1 lectin 21:26 A recombinant fungal lectin 0:26 A recombinant fungal lectin for labeling truncated glycans on human cancer cells 0:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 8 36 theme inhibitors 1127:1136 arg1 use 1120:1122 use 1120:1122 use of inhibitors of glycan metabolism 1120:1157 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 8 36 theme inhibitors 1127:1136 arg1 Labeling 1074:1081 Labeling 1074:1081 Labeling of several types of cancer cells 1074:1114 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 8 37 theme glycan 1141:1146 arg1 metabolism 1148:1157 glycan metabolism 1141:1157 glycan metabolism 1141:1157 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 6 38 theme Affinity 764:771 arg1 values 773:778 Affinity values 764:778 Affinity values obtained by microcalorimetry and surface plasmon resonance 764:837 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 2 39 theme healthy 360:366 arg1 tissues 368:374 healthy tissues 360:374 healthy tissues 360:374 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 0 40 theme truncated 41:49 arg1 glycans 51:57 truncated glycans 41:57 truncated glycans on human cancer cells 41:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 11 41 theme cancer 1597:1602 arg1 regions 1604:1610 cancer regions 1597:1610 cancer regions 1597:1610 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 10 42 theme cancer 1372:1377 arg1 tissue 1379:1384 cancer tissue 1372:1384 cancer tissue 1372:1384 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 7 43 theme Crystal 967:973 arg1 structure 975:983 Crystal structure 967:983 Crystal structure of PVL complexed with GlcNAcβ1-3Gal 967:1019 Crystal structure of PVL complexed with GlcNAcβ1-3Gal established the structural basis of the specificity. 26042789 6 44 theme micromolar 854:863 arg1 affinity 865:872 a micromolar affinity 852:872 a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches 852:964 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 10 45 theme healthy 1426:1432 arg1 ones 1434:1437 healthy ones 1426:1437 healthy ones 1426:1437 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 9 46 from amount 1302:1307 arg1 cells 1337:1341 cancer cells 1330:1341 cancer cells 1330:1341 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 6 47 theme GlcNAcβ1-2Man 936:948 arg1 branches 957:964 GlcNAcβ1-2Man capped branches 936:964 GlcNAcβ1-2Man capped branches 936:964 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 4 48 theme Psathyrella 602:612 arg1 PVL 624:626 PVL 624:626 PVL 624:626 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 4 48 theme Psathyrella 602:612 arg1 velutina 614:621 the fungus Psathyrella velutina 591:621 the fungus Psathyrella velutina (PVL) 591:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 6 49 theme surface 813:819 arg1 resonance 829:837 surface plasmon resonance 813:837 surface plasmon resonance 813:837 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 2 50 theme non-reducing 290:301 arg1 residues 334:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues 281:341 terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues 281:374 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 8 51 theme sialic 1216:1221 arg1 Neu5Ac 1229:1234 Neu5Ac 1229:1234 Neu5Ac 1229:1234 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 8 51 theme sialic 1216:1221 arg1 acid 1223:1226 sialic acid 1216:1226 sialic acid (Neu5Ac) 1216:1235 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 4 52 from yield 655:659 arg1 culture 674:680 bacterial culture 664:680 bacterial culture 664:680 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 1 53 theme oligosaccharide 184:198 arg1 epitopes 200:207 distinct oligosaccharide epitopes 175:207 distinct oligosaccharide epitopes 175:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 4 54 theme bacterial 664:672 arg1 culture 674:680 bacterial culture 664:680 bacterial culture 664:680 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 9 55 theme expression 1270:1279 arg1 Analysis 1238:1245 Analysis 1238:1245 Analysis of glycosyltransferase expression 1238:1279 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 11 56 located observed 1580:1587 arg1 carcinomas 1539:1548 lung, breast and colon carcinomas 1516:1548 lung, breast and colon carcinomas 1516:1548 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 11 56 located observed 1580:1587 arg2 delineation 1559:1569 a clear delineation 1551:1569 a clear delineation 1551:1569 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 5 57 theme GlcNAc 717:722 arg1 residues 724:731 terminal GlcNAc residues 708:731 terminal GlcNAc residues 708:731 A strong specificity for terminal GlcNAc residues was evidenced by glycan array. 26042789 11 58 theme breast 1522:1527 arg1 carcinomas 1539:1548 lung, breast and colon carcinomas 1516:1548 lung, breast and colon carcinomas 1516:1548 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 5 59 theme strong 685:690 arg1 specificity 692:702 A strong specificity 683:702 A strong specificity for terminal GlcNAc residues 683:731 A strong specificity for terminal GlcNAc residues was evidenced by glycan array. 26042789 9 60 theme GlcNAc 1312:1317 arg1 present 1319:1325 GlcNAc present 1312:1325 GlcNAc present 1312:1325 Analysis of glycosyltransferase expression confirmed the higher amount of GlcNAc present on cancer cells. 26042789 11 61 theme surrounding 1616:1626 arg1 tissues 1636:1642 surrounding healthy tissues 1616:1642 surrounding healthy tissues 1616:1642 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 11 62 theme lung 1516:1519 arg1 carcinomas 1539:1548 lung, breast and colon carcinomas 1516:1548 lung, breast and colon carcinomas 1516:1548 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 0 63 theme human 62:66 arg1 cells 75:79 human cancer cells 62:79 human cancer cells 62:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 2 64 theme truncated 255:263 arg1 glycans 265:271 truncated glycans 255:271 truncated glycans 255:271 Among them, truncated glycans present terminal non-reducing β-N-acetylglucosamine (GlcNAc) residues that are rare on healthy tissues. 26042789 1 65 from epitopes 200:207 arg1 diseases 162:169 chronic diseases 154:169 chronic diseases 154:169 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 1 66 theme structures 140:149 arg1 alterations 119:129 alterations 119:129 alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 119:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 1 67 dep glycoconjugates 95:109 arg1 present 111:117 present 111:117 present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 111:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 8 68 theme types 1094:1098 arg1 use 1120:1122 use 1120:1122 use of inhibitors of glycan metabolism 1120:1157 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 8 68 theme types 1094:1098 arg1 Labeling 1074:1081 Labeling 1074:1081 Labeling of several types of cancer cells 1074:1114 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 1 69 theme chronic 154:160 arg1 diseases 162:169 chronic diseases 154:169 chronic diseases 154:169 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 11 70 theme colon 1533:1537 arg1 carcinomas 1539:1548 lung, breast and colon carcinomas 1516:1548 lung, breast and colon carcinomas 1516:1548 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 8 71 theme cancer 1103:1108 arg1 cells 1110:1114 cancer cells 1103:1114 cancer cells 1103:1114 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 1 72 from diseases 162:169 arg1 alterations 119:129 alterations 119:129 alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 119:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 0 73 from glycans 51:57 arg1 cells 75:79 human cancer cells 62:79 human cancer cells 62:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 10 74 theme rPVL 1344:1347 arg1 binding 1349:1355 rPVL binding 1344:1355 rPVL binding 1344:1355 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 3 75 from sources 405:411 arg1 Lectins 377:383 Lectins 377:383 Lectins from unconventional sources such as fungi or algi 377:433 Lectins from unconventional sources such as fungi or algi provide novel markers that bind specifically to such epitopes, but their availability may be challenging. 26042789 12 76 theme labeling 1680:1687 arg1 agalacto-glycans 1689:1704 labeling agalacto-glycans 1680:1704 labeling agalacto-glycans in cancer or other diseases 1680:1732 PVL is therefore a useful tool for labeling agalacto-glycans in cancer or other diseases. 26042789 0 77 theme recombinant 2:12 arg1 lectin 21:26 A recombinant fungal lectin 0:26 A recombinant fungal lectin for labeling truncated glycans on human cancer cells 0:79 A recombinant fungal lectin for labeling truncated glycans on human cancer cells. 26042789 4 78 theme fruiting 574:581 arg1 body 583:586 the fruiting body 570:586 the fruiting body of the fungus Psathyrella velutina (PVL) 570:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 10 79 theme αGlcNAc-presenting 1486:1503 arg1 mucins 1505:1510 unique αGlcNAc-presenting mucins 1479:1510 unique αGlcNAc-presenting mucins 1479:1510 rPVL binding is specific to cancer tissue and weak or no labeling is observed for healthy ones, except for stomach glands that present unique αGlcNAc-presenting mucins. 26042789 1 80 from alterations 119:129 arg1 diseases 162:169 chronic diseases 154:169 chronic diseases 154:169 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 1 81 theme surface 87:93 arg1 glycoconjugates 95:109 Cell surface glycoconjugates 82:109 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes 82:207 Cell surface glycoconjugates present alterations of their structures in chronic diseases and distinct oligosaccharide epitopes have been associated with cancer. 26042789 8 82 theme metabolism 1148:1157 arg1 inhibitors 1127:1136 inhibitors 1127:1136 inhibitors of glycan metabolism 1127:1157 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 4 83 theme GlcNAc-binding 543:556 arg1 lectin 558:563 A GlcNAc-binding lectin 541:563 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) 541:627 A GlcNAc-binding lectin from the fruiting body of the fungus Psathyrella velutina (PVL) has been produced in good yield in bacterial culture. 26042789 11 84 theme healthy 1628:1634 arg1 tissues 1636:1642 surrounding healthy tissues 1616:1642 surrounding healthy tissues 1616:1642 In lung, breast and colon carcinomas, a clear delineation could be observed between cancer regions and surrounding healthy tissues. 26042789 6 85 theme GlcNAcβ1-3Gal 878:890 arg1 epitopes 892:899 GlcNAcβ1-3Gal epitopes 878:899 GlcNAcβ1-3Gal epitopes 878:899 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 26042789 8 86 theme terminal 1188:1195 arg1 GlcNAc 1197:1202 terminal GlcNAc 1188:1202 terminal GlcNAc 1188:1202 Labeling of several types of cancer cells and use of inhibitors of glycan metabolism indicated that rPVL binds to terminal GlcNAc but also to sialic acid (Neu5Ac). 26042789 6 87 with N-glycans 921:929 arg1 branches 957:964 GlcNAcβ1-2Man capped branches 936:964 GlcNAcβ1-2Man capped branches 936:964 Affinity values obtained by microcalorimetry and surface plasmon resonance demonstrated a micromolar affinity for GlcNAcβ1-3Gal epitopes and for biantennary N-glycans with GlcNAcβ1-2Man capped branches. 24212570 7 0 theme Such 1276:1279 arg1 demonstration 1281:1293 Such demonstration 1276:1293 Such demonstration of AglB substrate promiscuity 1276:1323 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 3 1 theme haloarchaeal 732:743 arg1 species 745:751 four haloarchaeal species 727:751 four haloarchaeal species 727:751 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 4 2 from glycans 826:832 arg1 hosts 850:854 their native hosts 837:854 their native hosts 837:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 7 3 theme substrate 1303:1311 arg1 promiscuity 1313:1323 AglB substrate promiscuity 1298:1323 AglB substrate promiscuity 1298:1323 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 2 4 theme lipid 487:491 arg1 carrier 493:499 the lipid carrier 483:499 the lipid carrier 483:499 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 7 5 gly N-glycosylation 1375:1389 arg1 elsewhere 1406:1414 elsewhere 1406:1414 elsewhere 1406:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 7 5 gly N-glycosylation 1375:1389 arg1 Archaea 1394:1400 Archaea 1394:1400 Archaea 1394:1400 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 2 6 theme composition 372:382 arg1 terms 350:354 terms 350:354 terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan 350:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 4 7 from salinarum 905:913 arg1 AglB 857:860 AglB 857:860 AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei 857:941 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 4 8 theme Haloferax 920:928 arg1 mediterranei 930:941 Haloferax mediterranei 920:941 Haloferax mediterranei 920:941 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 7 9 theme N-glycosylation 1375:1389 arg1 understanding 1358:1370 better understanding 1351:1370 better understanding of N-glycosylation in Archaea and elsewhere 1351:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 4 10 theme processing 793:802 arg1 glycans 826:832 processing distinct lipid-linked glycans 793:832 processing distinct lipid-linked glycans in their native hosts 793:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 7 11 from Archaea 1394:1400 arg1 understanding 1358:1370 better understanding 1351:1370 better understanding of N-glycosylation in Archaea and elsewhere 1351:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 6 12 theme enzymes 1267:1273 arg1 specificity 1246:1256 the relaxed substrate specificity 1224:1256 the relaxed substrate specificity of these enzymes 1224:1273 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 6 13 theme substrate 1236:1244 arg1 specificity 1246:1256 the relaxed substrate specificity 1224:1256 the relaxed substrate specificity of these enzymes 1224:1273 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 4 14 from volcanii 998:1005 arg1 counterpart 971:981 their counterpart 965:981 their counterpart from Haloferax volcanii 965:1005 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 2 15 theme composition 510:520 arg1 terms 350:354 terms 350:354 terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan 350:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 4 16 link lipid-linked 813:824 arg1 glycans 826:832 processing distinct lipid-linked glycans 793:832 processing distinct lipid-linked glycans in their native hosts 793:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 2 17 theme glycan 435:440 arg1 carrier 442:448 the lipid glycan carrier 425:448 the lipid glycan carrier 425:448 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 4 18 theme lipid-linked 813:824 arg1 glycans 826:832 processing distinct lipid-linked glycans 793:832 processing distinct lipid-linked glycans in their native hosts 793:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 0 19 theme lipid-linked 94:105 arg1 glycans 107:113 lipid-linked glycans 94:113 lipid-linked glycans 94:113 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 4 20 from marismortui 878:888 arg1 AglB 857:860 AglB 857:860 AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei 857:941 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 4 21 theme Haloarcula 867:876 arg1 marismortui 878:888 Haloarcula marismortui 867:888 Haloarcula marismortui 867:888 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 0 22 theme Substrate 0:8 arg1 promiscuity 10:20 Substrate promiscuity 0:20 Substrate promiscuity 0:20 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 0 22 theme Substrate 0:8 arg1 oligosaccharyltransferase 42:66 the archaeal oligosaccharyltransferase 29:66 the archaeal oligosaccharyltransferase 29:66 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 2 23 theme phosphorylation 406:420 arg1 composition 510:520 composition 510:520 composition 510:520 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 23 theme phosphorylation 406:420 arg1 composition 372:382 the chemical composition 359:382 the chemical composition 359:382 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 23 theme phosphorylation 406:420 arg1 structure 526:534 structure 526:534 structure 526:534 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 23 theme phosphorylation 406:420 arg1 sugar 455:459 the sugar 451:459 the sugar linking the glycan to the lipid carrier 451:499 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 23 theme phosphorylation 406:420 arg1 length 385:390 length 385:390 length 385:390 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 23 theme phosphorylation 406:420 arg1 degree 396:401 degree 396:401 degree 396:401 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 3 24 from species 745:751 arg1 AglB 717:720 the archaeal oligosaccharyltransferase AglB 678:720 the archaeal oligosaccharyltransferase AglB from four haloarchaeal species 678:751 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 4 25 theme Halobacterium 891:903 arg1 salinarum 905:913 Halobacterium salinarum 891:913 Halobacterium salinarum 891:913 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 6 26 theme structural 1126:1135 arg1 homology 1137:1144 significant sequence and apparently structural homology 1090:1144 homology 1137:1144 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 2 27 theme sugar 455:459 arg1 terms 350:354 terms 350:354 terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan 350:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 1 28 link lipid-linked 200:211 arg1 glycans 213:219 lipid-linked glycans 200:219 lipid-linked glycans 200:219 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 3 29 theme substrate 630:638 arg1 diversity 640:648 such substrate diversity 625:648 such substrate diversity 625:648 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 2 30 theme degree 396:401 arg1 terms 350:354 terms 350:354 terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan 350:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 4 31 theme native 843:848 arg1 hosts 850:854 their native hosts 837:854 their native hosts 837:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 2 32 theme glycan 555:560 arg1 composition 510:520 composition 510:520 composition 510:520 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 32 theme glycan 555:560 arg1 composition 372:382 the chemical composition 359:382 the chemical composition 359:382 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 32 theme glycan 555:560 arg1 structure 526:534 structure 526:534 structure 526:534 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 32 theme glycan 555:560 arg1 sugar 455:459 the sugar 451:459 the sugar linking the glycan to the lipid carrier 451:499 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 32 theme glycan 555:560 arg1 length 385:390 length 385:390 length 385:390 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 2 32 theme glycan 555:560 arg1 degree 396:401 degree 396:401 degree 396:401 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 6 33 theme AglB 1201:1204 arg1 proteins 1206:1213 the four AglB proteins 1192:1213 the four AglB proteins 1192:1213 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 1 34 theme selected 224:231 arg1 residues 237:244 selected Asn residues 224:244 selected Asn residues of target proteins 224:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 1 34 theme selected 224:231 arg1 proteins 256:263 target proteins 249:263 target proteins 249:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 6 35 theme proteins 1206:1213 arg1 similarity 1178:1187 the functional similarity 1163:1187 the functional similarity of the four AglB proteins 1163:1213 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 2 36 theme transferred 543:553 arg1 glycan 555:560 the transferred glycan 539:560 the transferred glycan 539:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 3 37 theme present 655:661 arg1 study 663:667 the present study 651:667 the present study 651:667 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 7 38 theme promiscuity 1313:1323 arg1 demonstration 1281:1293 Such demonstration 1276:1293 Such demonstration of AglB substrate promiscuity 1276:1323 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 1 39 theme Asn 233:235 arg1 residues 237:244 selected Asn residues 224:244 selected Asn residues of target proteins 224:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 1 39 theme Asn 233:235 arg1 proteins 256:263 target proteins 249:263 target proteins 249:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 4 40 theme Haloferax 988:996 arg1 volcanii 998:1005 Haloferax volcanii 988:1005 Haloferax volcanii 988:1005 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 6 41 theme significant 1090:1100 arg1 sequence 1102:1109 significant sequence and apparently structural homology 1090:1144 sequence 1102:1109 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 0 42 theme glycans 107:113 arg1 variety 83:89 a variety 81:89 a variety of lipid-linked glycans 81:113 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 0 42 theme glycans 107:113 arg1 glycans 107:113 lipid-linked glycans 94:113 lipid-linked glycans 94:113 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 2 43 theme lipid 429:433 arg1 carrier 442:448 the lipid glycan carrier 425:448 the lipid glycan carrier 425:448 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 7 44 from understanding 1358:1370 arg1 elsewhere 1406:1414 elsewhere 1406:1414 elsewhere 1406:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 7 44 from understanding 1358:1370 arg1 Archaea 1394:1400 Archaea 1394:1400 Archaea 1394:1400 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 8 45 theme glycoengineering 1480:1495 arg1 platform 1497:1504 a glycoengineering platform 1478:1504 a glycoengineering platform 1478:1504 volcanii into a glycoengineering platform. 24212570 2 46 theme chemical 363:370 arg1 composition 372:382 the chemical composition 359:382 the chemical composition 359:382 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 0 47 theme archaeal 33:40 arg1 promiscuity 10:20 Substrate promiscuity 0:20 Substrate promiscuity 0:20 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 0 47 theme archaeal 33:40 arg1 oligosaccharyltransferase 42:66 the archaeal oligosaccharyltransferase 29:66 the archaeal oligosaccharyltransferase 29:66 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 2 48 dep composition 510:520 arg1 the 506:508 the 506:508 the 506:508 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 3 49 theme archaeal 682:689 arg1 AglB 717:720 the archaeal oligosaccharyltransferase AglB 678:720 the archaeal oligosaccharyltransferase AglB from four haloarchaeal species 678:751 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 7 50 theme transforming 1446:1457 arg1 Hfx 1459:1461 transforming Hfx 1446:1461 transforming Hfx 1446:1461 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 7 51 from N-glycosylation 1375:1389 arg1 elsewhere 1406:1414 elsewhere 1406:1414 elsewhere 1406:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 7 51 from N-glycosylation 1375:1389 arg1 Archaea 1394:1400 Archaea 1394:1400 Archaea 1394:1400 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 7 52 theme better 1351:1356 arg1 understanding 1358:1370 better understanding 1351:1370 better understanding of N-glycosylation in Archaea and elsewhere 1351:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 3 53 theme oligosaccharyltransferase 691:715 arg1 AglB 717:720 the archaeal oligosaccharyltransferase AglB 678:720 the archaeal oligosaccharyltransferase AglB from four haloarchaeal species 678:751 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 2 54 theme similar 295:301 arg1 reactions 303:311 similar reactions 295:311 similar reactions 295:311 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 6 55 theme functional 1167:1176 arg1 similarity 1178:1187 the functional similarity 1163:1187 the functional similarity of the four AglB proteins 1163:1213 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 1 56 theme target 249:254 arg1 proteins 256:263 target proteins 249:263 target proteins 249:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 4 57 theme distinct 804:811 arg1 glycans 826:832 processing distinct lipid-linked glycans 793:832 processing distinct lipid-linked glycans in their native hosts 793:854 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 5 58 theme volcanii 1033:1040 arg1 cells 1042:1046 volcanii cells 1033:1046 volcanii cells 1033:1046 volcanii cells deleted of aglB. 24212570 6 59 theme relaxed 1228:1234 arg1 specificity 1246:1256 the relaxed substrate specificity 1224:1256 the relaxed substrate specificity of these enzymes 1224:1273 As the four enzymes show significant sequence and apparently structural homology, it appears that the functional similarity of the four AglB proteins reflects the relaxed substrate specificity of these enzymes. 24212570 2 60 theme carrier 442:448 arg1 phosphorylation 406:420 phosphorylation 406:420 phosphorylation of the lipid glycan carrier 406:448 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 24212570 1 61 theme proteins 256:263 arg1 residues 237:244 selected Asn residues 224:244 selected Asn residues of target proteins 224:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 1 61 theme proteins 256:263 arg1 proteins 256:263 target proteins 249:263 target proteins 249:263 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 4 62 from mediterranei 930:941 arg1 AglB 857:860 AglB 857:860 AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei 857:941 Accordingly, it was shown that despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui, Halobacterium salinarum, and Haloferax mediterranei could readily replace their counterpart from Haloferax volcanii when introduced into Hfx. 24212570 0 63 dep promiscuity 10:20 arg1 AglB 23:26 AglB 23:26 AglB 23:26 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 0 64 link lipid-linked 94:105 arg1 glycans 107:113 lipid-linked glycans 94:113 lipid-linked glycans 94:113 Substrate promiscuity: AglB, the archaeal oligosaccharyltransferase, can process a variety of lipid-linked glycans. 24212570 1 65 theme lipid-linked 200:211 arg1 glycans 213:219 lipid-linked glycans 200:219 lipid-linked glycans 200:219 Across evolution, N-glycosylation involves oligosaccharyltransferases that transfer lipid-linked glycans to selected Asn residues of target proteins. 24212570 7 66 theme AglB 1298:1301 arg1 promiscuity 1313:1323 AglB substrate promiscuity 1298:1323 AglB substrate promiscuity 1298:1323 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 3 67 theme such 625:628 arg1 diversity 640:648 such substrate diversity 625:648 such substrate diversity 625:648 To gain insight into how oligosaccharyltransferases cope with such substrate diversity, the present study analyzed the archaeal oligosaccharyltransferase AglB from four haloarchaeal species. 24212570 7 68 from elsewhere 1406:1414 arg1 understanding 1358:1370 better understanding 1351:1370 better understanding of N-glycosylation in Archaea and elsewhere 1351:1414 Such demonstration of AglB substrate promiscuity is important not only for better understanding of N-glycosylation in Archaea and elsewhere but also for efforts aimed at transforming Hfx. 24212570 2 69 theme length 385:390 arg1 terms 350:354 terms 350:354 terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan 350:560 While these enzymes catalyze similar reactions in each domain, differences exist in terms of the chemical composition, length and degree of phosphorylation of the lipid glycan carrier, the sugar linking the glycan to the lipid carrier, and the composition and structure of the transferred glycan. 27717196 3 0 theme whole 636:640 arg1 lysates 647:653 whole cell lysates 636:653 whole cell lysates (WCL) 636:659 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 3 0 theme whole 636:640 arg1 WCL 656:658 WCL 656:658 WCL 656:658 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 3 1 theme cell 642:645 arg1 lysates 647:653 whole cell lysates 636:653 whole cell lysates (WCL) 636:659 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 3 1 theme cell 642:645 arg1 WCL 656:658 WCL 656:658 WCL 656:658 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 0 2 theme acridone 72:79 arg1 derivatives 81:91 acridone derivatives 72:91 acridone derivatives 72:91 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 2 3 theme action 318:323 arg1 mechanism 325:333 the underlying action mechanism 303:333 the underlying action mechanism of 8a 303:339 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 3 4 theme isomer-sensitive 705:720 arg1 nano-LC/MS 759:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 0 5 from Changes 0:6 arg1 glycosylation 20:32 cellular glycosylation 11:32 cellular glycosylation of leukemia cells upon treatment with acridone derivatives 11:91 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 5 6 from changes 1016:1022 arg1 glycosylation 1027:1039 glycosylation 1027:1039 glycosylation 1027:1039 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 7 from indicator 1003:1011 arg1 glycosylation 1027:1039 glycosylation 1027:1039 glycosylation 1027:1039 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 8 8 used used 1433:1436 arg2 biomarkers 1450:1459 possible biomarkers 1441:1459 possible biomarkers for monitoring toxicity and antitumor activity of 8a 1441:1512 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 8 8 used used 1433:1436 arg2 glycosylation 1364:1376 glycosylation 1364:1376 glycosylation 1364:1376 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 0 9 theme drug 112:115 arg1 action 117:122 drug action 112:122 drug action 112:122 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 2 10 theme CCRF-CEM 507:514 arg1 cell 525:528 CCRF-CEM leukemia cell 507:528 CCRF-CEM leukemia cell 507:528 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 0 11 with treatment 57:65 arg1 derivatives 81:91 acridone derivatives 72:91 acridone derivatives 72:91 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 1 12 contain have 248:251 arg1 derivative 140:149 A new acridone derivative 125:149 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) 125:228 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 12 contain have 248:251 arg2 activity 270:277 potent antitumor activity 253:277 potent antitumor activity 253:277 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 7 13 theme functional 1225:1234 arg1 role 1236:1239 a key functional role 1219:1239 a key functional role 1219:1239 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 8 14 theme possible 1441:1448 arg1 biomarkers 1450:1459 possible biomarkers 1441:1459 possible biomarkers for monitoring toxicity and antitumor activity of 8a 1441:1512 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 8 14 theme possible 1441:1448 arg1 glycosylation 1364:1376 glycosylation 1364:1376 glycosylation 1364:1376 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 5 15 theme sensitive 993:1001 arg1 CM 976:977 CM 976:977 CM 976:977 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 15 theme sensitive 993:1001 arg1 indicator 1003:1011 a more sensitive indicator 986:1011 a more sensitive indicator of changes in glycosylation 986:1039 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 16 theme analyses 874:881 arg1 Comparison 852:861 Comparison 852:861 Comparison of the two analyses 852:881 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 4 17 theme compounds 825:833 arg1 total 773:777 A total 771:777 A total of 223 N-glycan compositions and 398 N-glycan compounds 771:833 A total of 223 N-glycan compositions and 398 N-glycan compounds were identified. 27717196 2 18 theme 10- 437:439 arg1 -acridone 469:477 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 18 theme 10- 437:439 arg1 -acridone 422:430 9(10H)-acridone 416:430 9(10H)-acridone (A) 416:434 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 19 with class 370:374 arg1 structures 381:390 structures 381:390 structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, 381:489 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 3 20 from lysates 647:653 arg1 N-Glycans 621:629 N-Glycans 621:629 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) 621:683 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 2 21 dep understand 292:301 arg1 to 289:290 to 289:290 to 289:290 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 1 22 theme 5-dimethoxy 175:185 arg1 8a 226:227 8a 226:227 8a 226:227 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 22 theme 5-dimethoxy 175:185 arg1 hydrochloride 211:223 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride 151:223 hydrochloride 211:223 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 2 23 theme step-by-step 402:413 arg1 A 433:433 A 433:433 A 433:433 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 23 theme step-by-step 402:413 arg1 -acridone 469:477 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 23 theme step-by-step 402:413 arg1 -acridone 422:430 9(10H)-acridone 416:430 9(10H)-acridone (A) 416:434 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 23 theme step-by-step 402:413 arg1 8a 487:488 8a 487:488 8a 487:488 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 24 theme N-glycosylation 547:561 arg1 changes 563:569 the N-glycosylation changes 543:569 the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform 543:618 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 7 25 theme 8a 1319:1320 arg1 mechanism 1306:1314 the action mechanism 1295:1314 the action mechanism of 8a 1295:1320 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 7 26 theme subunits 1195:1202 arg1 expressions 1154:1164 Different expressions 1144:1164 Different expressions of oligosaccharyltransferase subunits 1144:1202 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 7 27 theme key 1221:1223 arg1 role 1236:1239 a key functional role 1219:1239 a key functional role 1219:1239 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 2 28 theme leukemia 516:523 arg1 cell 525:528 CCRF-CEM leukemia cell 507:528 CCRF-CEM leukemia cell 507:528 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 7 29 theme action 1299:1304 arg1 mechanism 1306:1314 the action mechanism 1295:1314 the action mechanism of 8a 1295:1320 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 6 30 theme high-mannose-type 1104:1120 arg1 glycans 1122:1128 high-mannose-type glycans 1104:1128 high-mannose-type glycans 1104:1128 Upon 8a exposure to CCRF-CEM cells, a significant decrease in high-mannose-type glycans was observed. 27717196 1 31 theme -benzyl-9 187:195 arg1 8a 226:227 8a 226:227 8a 226:227 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 31 theme -benzyl-9 187:195 arg1 hydrochloride 211:223 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride 151:223 hydrochloride 211:223 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 0 32 theme cellular 11:18 arg1 glycosylation 20:32 cellular glycosylation 11:32 cellular glycosylation of leukemia cells upon treatment with acridone derivatives 11:91 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 1 33 theme potent 253:258 arg1 activity 270:277 potent antitumor activity 253:277 potent antitumor activity 253:277 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 2 34 theme 8a 338:339 arg1 mechanism 325:333 the underlying action mechanism 303:333 the underlying action mechanism of 8a 303:339 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 1 35 dep derivative 140:149 arg1 8a 226:227 8a 226:227 8a 226:227 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 35 dep derivative 140:149 arg1 hydrochloride 211:223 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride 151:223 hydrochloride 211:223 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 2 36 theme class 370:374 arg1 compounds 348:356 three compounds 342:356 three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, 342:489 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 0 37 theme leukemia 37:44 arg1 cells 46:50 leukemia cells 37:50 leukemia cells 37:50 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 1 38 theme antitumor 260:268 arg1 activity 270:277 potent antitumor activity 253:277 potent antitumor activity 253:277 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 2 39 theme benzyl-9 456:463 arg1 -acridone 469:477 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) 437:481 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 39 theme benzyl-9 456:463 arg1 -acridone 422:430 9(10H)-acridone 416:430 9(10H)-acridone (A) 416:434 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 40 theme same 365:368 arg1 class 370:374 the same class 361:374 the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, 361:489 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 5 41 from glycosylation 1027:1039 arg1 CM 976:977 CM 976:977 CM 976:977 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 41 from glycosylation 1027:1039 arg1 indicator 1003:1011 a more sensitive indicator 986:1011 a more sensitive indicator of changes in glycosylation 986:1039 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 1 42 theme 10H 197:199 arg1 8a 226:227 8a 226:227 8a 226:227 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 42 theme 10H 197:199 arg1 hydrochloride 211:223 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride 151:223 hydrochloride 211:223 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 6 43 from decrease 1092:1099 arg1 glycans 1122:1128 high-mannose-type glycans 1104:1128 high-mannose-type glycans 1104:1128 Upon 8a exposure to CCRF-CEM cells, a significant decrease in high-mannose-type glycans was observed. 27717196 5 44 theme changes 1016:1022 arg1 CM 976:977 CM 976:977 CM 976:977 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 44 theme changes 1016:1022 arg1 indicator 1003:1011 a more sensitive indicator 986:1011 a more sensitive indicator of changes in glycosylation 986:1039 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 4 45 theme N-glycan 816:823 arg1 compounds 825:833 398 N-glycan compounds 812:833 398 N-glycan compounds 812:833 A total of 223 N-glycan compositions and 398 N-glycan compounds were identified. 27717196 2 46 theme MS 608:609 arg1 platform 611:618 the microfluidic HPLC-chip-TOF MS platform 577:618 the microfluidic HPLC-chip-TOF MS platform 577:618 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 3 47 theme cell 665:668 arg1 CM 681:682 CM 681:682 CM 681:682 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 3 47 theme cell 665:668 arg1 membranes 670:678 cell membranes 665:678 cell membranes (CM) 665:683 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 3 48 from membranes 670:678 arg1 N-Glycans 621:629 N-Glycans 621:629 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) 621:683 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 8 49 theme therapeutic 1402:1412 arg1 potency 1414:1420 therapeutic potency 1402:1420 therapeutic potency 1402:1420 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 2 50 theme HPLC-chip-TOF 594:606 arg1 platform 611:618 the microfluidic HPLC-chip-TOF MS platform 577:618 the microfluidic HPLC-chip-TOF MS platform 577:618 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 2 51 theme underlying 307:316 arg1 mechanism 325:333 the underlying action mechanism 303:333 the underlying action mechanism of 8a 303:339 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 0 52 theme cells 46:50 arg1 glycosylation 20:32 cellular glycosylation 11:32 cellular glycosylation of leukemia cells upon treatment with acridone derivatives 11:91 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 8 53 dep suggest 1351:1357 arg1 Taken 1323:1327 Taken 1323:1327 Taken together 1323:1336 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 0 54 gly glycosylation 20:32 arg1 cells 46:50 leukemia cells 37:50 leukemia cells 37:50 Changes in cellular glycosylation of leukemia cells upon treatment with acridone derivatives yield insight into drug action. 27717196 2 55 theme microfluidic 581:592 arg1 platform 611:618 the microfluidic HPLC-chip-TOF MS platform 577:618 the microfluidic HPLC-chip-TOF MS platform 577:618 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 7 56 theme oligosaccharyltransferase 1169:1193 arg1 subunits 1195:1202 oligosaccharyltransferase subunits 1169:1202 oligosaccharyltransferase subunits 1169:1202 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 3 57 theme graphitized 740:750 arg1 nano-LC/MS 759:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 6 58 theme CCRF-CEM 1062:1069 arg1 cells 1071:1075 CCRF-CEM cells 1062:1075 CCRF-CEM cells 1062:1075 Upon 8a exposure to CCRF-CEM cells, a significant decrease in high-mannose-type glycans was observed. 27717196 4 59 theme compositions 795:806 arg1 total 773:777 A total 771:777 A total of 223 N-glycan compositions and 398 N-glycan compounds 771:833 A total of 223 N-glycan compositions and 398 N-glycan compounds were identified. 27717196 6 60 theme 8a 1047:1048 arg1 exposure 1050:1057 8a exposure 1047:1057 8a exposure to CCRF-CEM cells 1047:1075 Upon 8a exposure to CCRF-CEM cells, a significant decrease in high-mannose-type glycans was observed. 27717196 7 61 theme Different 1144:1152 arg1 expressions 1154:1164 Different expressions 1144:1164 Different expressions of oligosaccharyltransferase subunits 1144:1202 Different expressions of oligosaccharyltransferase subunits appear to play a key functional role in regulating the hypoglycosylation and contribute to the action mechanism of 8a. 27717196 3 62 theme carbon 752:757 arg1 nano-LC/MS 759:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 1 63 theme -acridone 201:209 arg1 8a 226:227 8a 226:227 8a 226:227 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 1 63 theme -acridone 201:209 arg1 hydrochloride 211:223 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride 151:223 hydrochloride 211:223 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 4 64 theme N-glycan 786:793 arg1 compositions 795:806 223 N-glycan compositions 782:806 223 N-glycan compositions 782:806 A total of 223 N-glycan compositions and 398 N-glycan compounds were identified. 27717196 1 65 theme new 127:129 arg1 derivative 140:149 A new acridone derivative 125:149 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) 125:228 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 3 66 theme chip-based 722:731 arg1 nano-LC/MS 759:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 8 67 theme 8a 1511:1512 arg1 toxicity 1476:1483 toxicity 1476:1483 toxicity 1476:1483 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 8 67 theme 8a 1511:1512 arg1 activity 1499:1506 antitumor activity 1489:1506 antitumor activity 1489:1506 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 2 68 dep -acridone 469:477 arg1 10H 465:467 10H 465:467 10H 465:467 In order to understand the underlying action mechanism of 8a, three compounds of the same class with structures optimized step-by-step, 9(10H)-acridone (A), 10-(3,5-dimethoxy) benzyl-9(10H)-acridone (I) and 8a, were exposed to CCRF-CEM leukemia cell to determine the N-glycosylation changes using the microfluidic HPLC-chip-TOF MS platform. 27717196 1 69 theme acridone 131:138 arg1 derivative 140:149 A new acridone derivative 125:149 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) 125:228 A new acridone derivative 2-aminoacetamido-10-(3, 5-dimethoxy)-benzyl-9(10H)-acridone hydrochloride (8a) has been shown to have potent antitumor activity. 27717196 3 70 theme porous 733:738 arg1 nano-LC/MS 759:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 isomer-sensitive chip-based porous graphitized carbon nano-LC/MS 705:768 N-Glycans from whole cell lysates (WCL) and cell membranes (CM) were analyzed using isomer-sensitive chip-based porous graphitized carbon nano-LC/MS. 27717196 8 71 theme antitumor 1489:1497 arg1 activity 1499:1506 antitumor activity 1489:1506 antitumor activity 1489:1506 Taken together our findings suggest that glycosylation is strongly affected by therapeutic potency and can be used as possible biomarkers for monitoring toxicity and antitumor activity of 8a. 27717196 5 72 theme apparent 900:907 arg1 changes 909:915 more apparent changes 895:915 more apparent changes 895:915 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 6 73 theme significant 1080:1090 arg1 decrease 1092:1099 a significant decrease 1078:1099 a significant decrease in high-mannose-type glycans 1078:1128 Upon 8a exposure to CCRF-CEM cells, a significant decrease in high-mannose-type glycans was observed. 27717196 5 74 located observed 922:929 arg1 CM 938:939 the CM 934:939 the CM compared with WCL 934:957 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 27717196 5 74 located observed 922:929 arg2 changes 909:915 more apparent changes 895:915 more apparent changes 895:915 Comparison of the two analyses showed that more apparent changes were observed in the CM compared with WCL, suggesting that CM may be a more sensitive indicator of changes in glycosylation. 28895795 0 0 theme analytical 73:82 arg1 solution 84:91 a total analytical solution: 65:92 a total analytical solution: From sample preparation to data analysis 65:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 3 1 theme entire 534:539 arg1 process 541:547 the entire process 530:547 the entire process from sample preparation to data analysis 530:588 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 2 2 with process 326:332 arg1 protocols 363:371 long sample preparation protocols 339:371 long sample preparation protocols 339:371 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 2 2 with process 326:332 arg1 interpretation 384:397 manual interpretation 377:397 manual interpretation of the data 377:409 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 0 3 theme total 67:71 arg1 solution 84:91 a total analytical solution: 65:92 a total analytical solution: From sample preparation to data analysis 65:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 6 4 theme structural 1067:1076 arg1 assignment 1078:1087 structural assignment 1067:1087 structural assignment 1067:1087 Following HILIC-UPLC/FLR/MS analyses, the data are processed and a library search based on glucose units has been included to expedite the task of structural assignment. 28895795 2 5 theme manual 377:382 arg1 interpretation 384:397 manual interpretation 377:397 manual interpretation of the data 377:409 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 6 6 theme HILIC-UPLC/FLR/MS 930:946 arg1 analyses 948:955 HILIC-UPLC/FLR/MS analyses 930:955 HILIC-UPLC/FLR/MS analyses 930:955 Following HILIC-UPLC/FLR/MS analyses, the data are processed and a library search based on glucose units has been included to expedite the task of structural assignment. 28895795 3 7 theme data 576:579 arg1 analysis 581:588 data analysis 576:588 data analysis 576:588 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 4 8 theme fast 693:696 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 6 9 theme glucose 1011:1017 arg1 units 1019:1023 glucose units 1011:1023 glucose units 1011:1023 Following HILIC-UPLC/FLR/MS analyses, the data are processed and a library search based on glucose units has been included to expedite the task of structural assignment. 28895795 9 10 theme glycans 1379:1385 arg1 neutral 1392:1398 neutral 1392:1398 neutral 1392:1398 The majority of the glycans were neutral and fucosylated; glycans featuring N-glycolylneuraminic acid and those with two galactoses connected via an α1,3-linkage were also identified. 28895795 9 10 theme glycans 1379:1385 arg1 majority 1363:1370 The majority 1359:1370 The majority of the glycans 1359:1385 The majority of the glycans were neutral and fucosylated; glycans featuring N-glycolylneuraminic acid and those with two galactoses connected via an α1,3-linkage were also identified. 28895795 5 11 theme mass 886:889 arg1 sensitivities 905:917 mass spectral (MS) sensitivities 886:917 mass spectral (MS) sensitivities 886:917 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 0 12 theme sample 99:104 arg1 preparation 106:116 sample preparation 99:116 sample preparation 99:116 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 7 13 theme total 1111:1115 arg1 solution 1128:1135 this total analytical solution 1106:1135 this total analytical solution 1106:1135 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 8 14 gly glycoprotein 1221:1232 arg1 glycoprotein 1221:1232 this glycoprotein 1216:1232 this glycoprotein 1216:1232 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 5 15 with labelling 815:823 arg1 tag 843:845 RapiFluor-MS tag 830:845 RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities 830:917 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 4 16 theme glycan 707:712 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 1 17 theme important 156:164 arg1 Glycosylation 136:148 Glycosylation 136:148 Glycosylation 136:148 Glycosylation is an important attribute of biopharmaceutical products to monitor from development through production. 28895795 1 17 theme important 156:164 arg1 attribute 166:174 an important attribute 153:174 an important attribute of biopharmaceutical products to monitor from development through production 153:251 Glycosylation is an important attribute of biopharmaceutical products to monitor from development through production. 28895795 7 18 theme analytical 1117:1126 arg1 solution 1128:1135 this total analytical solution 1106:1135 this total analytical solution 1106:1135 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 7 19 theme 8761 1206:1209 arg1 glycosylation 1157:1169 the glycosylation 1153:1169 the glycosylation of the NIST Reference Material mAb 8761 1153:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 5 20 theme spectral 891:898 arg1 sensitivities 905:917 mass spectral (MS) sensitivities 886:917 mass spectral (MS) sensitivities 886:917 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 7 21 theme mAb 1202:1204 arg1 8761 1206:1209 the NIST Reference Material mAb 8761 1174:1209 the NIST Reference Material mAb 8761 1174:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 2 22 theme long 339:342 arg1 protocols 363:371 long sample preparation protocols 339:371 long sample preparation protocols 339:371 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 3 23 theme glycan 454:459 arg1 analysis 461:468 glycan analysis 454:468 glycan analysis 454:468 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 0 24 theme Glycan 0:5 arg1 characterization 7:22 Glycan characterization 0:22 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 0:134 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 7 25 theme NIST 1178:1181 arg1 8761 1206:1209 the NIST Reference Material mAb 8761 1174:1209 the NIST Reference Material mAb 8761 1174:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 8 26 theme N-linked 1264:1271 arg1 glycans 1273:1279 35 N-linked glycans 1261:1279 35 N-linked glycans 1261:1279 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 2 27 theme data 406:409 arg1 protocols 363:371 long sample preparation protocols 339:371 long sample preparation protocols 339:371 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 2 27 theme data 406:409 arg1 interpretation 384:397 manual interpretation 377:397 manual interpretation of the data 377:409 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 7 28 theme Reference 1183:1191 arg1 8761 1206:1209 the NIST Reference Material mAb 8761 1174:1209 the NIST Reference Material mAb 8761 1174:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 9 29 theme N-glycolylneuraminic 1435:1454 arg1 acid 1456:1459 N-glycolylneuraminic acid 1435:1459 N-glycolylneuraminic acid 1435:1459 The majority of the glycans were neutral and fucosylated; glycans featuring N-glycolylneuraminic acid and those with two galactoses connected via an α1,3-linkage were also identified. 28895795 1 30 theme products 197:204 arg1 Glycosylation 136:148 Glycosylation 136:148 Glycosylation 136:148 Glycosylation is an important attribute of biopharmaceutical products to monitor from development through production. 28895795 1 30 theme products 197:204 arg1 attribute 166:174 an important attribute 153:174 an important attribute of biopharmaceutical products to monitor from development through production 153:251 Glycosylation is an important attribute of biopharmaceutical products to monitor from development through production. 28895795 5 31 theme fluorescence 863:874 arg1 FLR 877:879 fluorescence (FLR) 863:880 fluorescence (FLR) 863:880 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 6 32 theme library 987:993 arg1 search 995:1000 a library search 985:1000 a library search based on glucose units 985:1023 Following HILIC-UPLC/FLR/MS analyses, the data are processed and a library search based on glucose units has been included to expedite the task of structural assignment. 28895795 2 33 theme time-consuming 311:324 arg1 process 326:332 a time-consuming process 309:332 a time-consuming process 309:332 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 2 33 theme time-consuming 311:324 arg1 analysis 277:284 glycosylation analysis 263:284 glycosylation analysis 263:284 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 3 34 theme analytical 498:507 arg1 solution 509:516 a streamlined analytical solution 484:516 a streamlined analytical solution that covers the entire process from sample preparation to data analysis 484:588 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 0 35 theme RM 36:37 arg1 antibody 50:57 the NIST RM monoclonal antibody 27:57 the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis 27:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 1 36 theme biopharmaceutical 179:195 arg1 products 197:204 biopharmaceutical products 179:204 biopharmaceutical products 179:204 Glycosylation is an important attribute of biopharmaceutical products to monitor from development through production. 28895795 4 37 theme simplified 678:687 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 0 38 theme NIST 31:34 arg1 antibody 50:57 the NIST RM monoclonal antibody 27:57 the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis 27:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 0 39 theme data 121:124 arg1 analysis 126:133 data analysis 121:133 data analysis 121:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 8 40 link N-linked 1264:1271 arg1 glycans 1273:1279 35 N-linked glycans 1261:1279 35 N-linked glycans 1261:1279 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 4 41 link N-linked 698:705 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 8 42 theme major 1295:1299 arg1 mannose 1315:1321 high mannose 1310:1321 high mannose 1310:1321 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 8 42 theme major 1295:1299 arg1 hybrid 1337:1342 hybrid 1337:1342 hybrid 1337:1342 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 8 42 theme major 1295:1299 arg1 complex 1324:1330 complex 1324:1330 complex 1324:1330 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 8 42 theme major 1295:1299 arg1 classes 1301:1307 all three major classes 1285:1307 all three major classes 1285:1307 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 0 43 theme antibody 50:57 arg1 characterization 7:22 Glycan characterization 0:22 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 0:134 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 5 44 theme sample 787:792 arg1 preparation 794:804 The sample preparation 783:804 The sample preparation 783:804 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 8 45 theme high 1310:1313 arg1 mannose 1315:1321 high mannose 1310:1321 high mannose 1310:1321 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 8 45 theme high 1310:1313 arg1 classes 1301:1307 all three major classes 1285:1307 all three major classes 1285:1307 For this glycoprotein, we confidently identified 35 N-linked glycans and all three major classes, high mannose, complex, and hybrid, were present. 28895795 0 46 theme monoclonal 39:48 arg1 antibody 50:57 the NIST RM monoclonal antibody 27:57 the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis 27:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28895795 6 47 theme assignment 1078:1087 arg1 task 1059:1062 the task 1055:1062 the task of structural assignment 1055:1087 Following HILIC-UPLC/FLR/MS analyses, the data are processed and a library search based on glucose units has been included to expedite the task of structural assignment. 28895795 7 48 gly glycosylation 1157:1169 arg1 8761 1206:1209 the NIST Reference Material mAb 8761 1174:1209 the NIST Reference Material mAb 8761 1174:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 4 49 theme N-linked 698:705 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 4 50 theme sample 714:719 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 2 51 theme preparation 351:361 arg1 protocols 363:371 long sample preparation protocols 339:371 long sample preparation protocols 339:371 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 5 52 theme MS 901:902 arg1 sensitivities 905:917 mass spectral (MS) sensitivities 886:917 mass spectral (MS) sensitivities 886:917 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 4 53 theme analytical 639:648 arg1 solution 650:657 the complete analytical solution 626:657 the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 626:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 2 54 theme glycosylation 263:275 arg1 process 326:332 a time-consuming process 309:332 a time-consuming process 309:332 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 2 54 theme glycosylation 263:275 arg1 analysis 277:284 glycosylation analysis 263:284 glycosylation analysis 263:284 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 2 55 theme sample 344:349 arg1 protocols 363:371 long sample preparation protocols 339:371 long sample preparation protocols 339:371 However, glycosylation analysis has traditionally been a time-consuming process with long sample preparation protocols and manual interpretation of the data. 28895795 4 56 theme complete 630:637 arg1 solution 650:657 the complete analytical solution 626:657 the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 626:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 3 57 theme streamlined 486:496 arg1 solution 509:516 a streamlined analytical solution 484:516 a streamlined analytical solution that covers the entire process from sample preparation to data analysis 484:588 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 4 58 theme preparation 721:731 arg1 protocol 733:740 a simplified and fast N-linked glycan sample preparation protocol 676:740 a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr 676:780 In this communication, we describe the complete analytical solution that begins with a simplified and fast N-linked glycan sample preparation protocol that can be completed in less than 1 hr. 28895795 5 59 theme RapiFluor-MS 830:841 arg1 tag 843:845 RapiFluor-MS tag 830:845 RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities 830:917 The sample preparation includes labelling with RapiFluor-MS tag to improve both fluorescence (FLR) and mass spectral (MS) sensitivities. 28895795 7 60 theme Material 1193:1200 arg1 8761 1206:1209 the NIST Reference Material mAb 8761 1174:1209 the NIST Reference Material mAb 8761 1174:1209 We then applied this total analytical solution to characterize the glycosylation of the NIST Reference Material mAb 8761. 28895795 3 61 theme sample 554:559 arg1 preparation 561:571 sample preparation 554:571 sample preparation 554:571 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 3 62 from preparation 561:571 arg1 process 541:547 the entire process 530:547 the entire process from sample preparation to data analysis 530:588 To address the challenges associated with glycan analysis, we developed a streamlined analytical solution that covers the entire process from sample preparation to data analysis. 28895795 0 63 from preparation 106:116 arg1 solution 84:91 a total analytical solution: 65:92 a total analytical solution: From sample preparation to data analysis 65:133 Glycan characterization of the NIST RM monoclonal antibody using a total analytical solution: From sample preparation to data analysis. 28231413 1 0 theme cells 380:384 arg1 surface 362:368 the surface 358:368 the surface of living cells 358:384 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 5 1 from biosynthesis 902:913 arg1 cells 938:942 cells 938:942 cells 938:942 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 1 2 theme structural 176:185 arg1 flexibility 187:197 structural flexibility 176:197 structural flexibility 176:197 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 1 3 theme unnatural 290:298 arg1 analogues 315:323 unnatural monosaccharide analogues 290:323 unnatural monosaccharide analogues 290:323 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 6 4 theme different 1078:1086 arg1 sugars 1094:1099 the different donor sugars 1074:1099 the different donor sugars 1074:1099 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 6 5 dep so 1159:1160 arg1 less 1154:1157 less 1154:1157 less 1154:1157 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 5 6 theme N-acetylglucosamine- 986:1005 arg1 MCRs 1039:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 1 7 theme monosaccharide 300:313 arg1 analogues 315:323 unnatural monosaccharide analogues 290:323 unnatural monosaccharide analogues 290:323 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 5 8 theme N-acetylgalactosamine-based 1011:1037 arg1 MCRs 1039:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 5 9 theme MCRs 1039:1042 arg1 relationship 877:888 the relationship 873:888 the relationship between the biosynthesis of MCR donor sugars in cells 873:942 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 5 9 theme MCRs 1039:1042 arg1 levels 961:966 the labeling levels 948:966 the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 948:1042 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 5 10 theme labeling 952:959 arg1 levels 961:966 the labeling levels 948:966 the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 948:1042 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 6 11 theme labeling 1134:1141 arg1 levels 1143:1148 the overall labeling levels 1122:1148 the overall labeling levels 1122:1148 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 0 12 theme Cell-Type-Selective 48:66 arg1 Metabolism 68:77 Cell-Type-Selective Metabolism 48:77 Cell-Type-Selective Metabolism 48:77 Metabolic Chemical Reporters of Glycans Exhibit Cell-Type-Selective Metabolism and Glycoprotein Labeling. 28231413 6 13 theme intracellular 1167:1179 arg1 labeling 1181:1188 intracellular labeling 1167:1188 intracellular labeling of proteins by O-GlcNAcylation 1167:1219 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 2 14 theme visualization 502:514 arg1 installation 486:497 the selective installation 472:497 the selective installation of visualization or enrichment tags 472:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 2 15 theme bioorthogonal 439:451 arg1 groups 453:458 bioorthogonal groups 439:458 bioorthogonal groups that enable the selective installation of visualization or enrichment tags 439:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 0 16 theme Chemical 10:17 arg1 Reporters 19:27 Metabolic Chemical Reporters 0:27 Metabolic Chemical Reporters of Glycans 0:38 Metabolic Chemical Reporters of Glycans Exhibit Cell-Type-Selective Metabolism and Glycoprotein Labeling. 28231413 5 17 theme different 976:984 arg1 MCRs 1039:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs 971:1042 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 1 18 theme carbohydrate 206:217 arg1 biosynthesis 219:230 the carbohydrate biosynthesis and glycosylation pathways 202:257 biosynthesis 219:230 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 0 19 theme Metabolic 0:8 arg1 Reporters 19:27 Metabolic Chemical Reporters 0:27 Metabolic Chemical Reporters of Glycans 0:38 Metabolic Chemical Reporters of Glycans Exhibit Cell-Type-Selective Metabolism and Glycoprotein Labeling. 28231413 2 20 theme monosaccharides 415:429 arg1 subset 389:394 A subset 387:394 A subset of these unnatural monosaccharides 387:429 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 6 21 theme proteins 1193:1200 arg1 labeling 1181:1188 intracellular labeling 1167:1188 intracellular labeling of proteins by O-GlcNAcylation 1167:1219 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 1 22 theme pioneering 116:125 arg1 work 127:130 the pioneering work 112:130 the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways 112:257 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 2 23 theme selective 476:484 arg1 installation 486:497 the selective installation 472:497 the selective installation of visualization or enrichment tags 472:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 1 24 theme glycan 336:341 arg1 engineering 343:353 glycan engineering 336:353 glycan engineering 336:353 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 3 25 theme unbiased 613:620 arg1 identification 622:635 the unbiased identification 609:635 the unbiased identification of glycoproteins 609:652 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 1 26 theme glycosylation 236:248 arg1 pathways 250:257 the carbohydrate biosynthesis and glycosylation pathways 202:257 pathways 250:257 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 4 27 gly glycoprotein 842:853 arg1 glycoprotein 842:853 glycoprotein 842:853 glycoprotein 842:853 For example, they are incorporated substoichiometrically into glycans, and most MCRs are not selective for one class (e.g., O-GlcNAcylation) of glycoprotein. 28231413 2 28 contain contain 431:437 arg2 groups 453:458 bioorthogonal groups 439:458 bioorthogonal groups that enable the selective installation of visualization or enrichment tags 439:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 2 28 contain contain 431:437 arg1 subset 389:394 A subset 387:394 A subset of these unnatural monosaccharides 387:429 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 4 29 dep O-GlcNAcylation 822:836 arg1 e.g. 816:819 e.g. 816:819 e.g. 816:819 For example, they are incorporated substoichiometrically into glycans, and most MCRs are not selective for one class (e.g., O-GlcNAcylation) of glycoprotein. 28231413 6 30 theme sugars 1094:1099 arg1 buildup 1063:1069 the buildup 1059:1069 the buildup of the different donor sugars 1059:1099 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 0 31 theme Glycans 32:38 arg1 Reporters 19:27 Metabolic Chemical Reporters 0:27 Metabolic Chemical Reporters of Glycans 0:38 Metabolic Chemical Reporters of Glycans Exhibit Cell-Type-Selective Metabolism and Glycoprotein Labeling. 28231413 3 32 gly glycoproteins 640:652 arg1 glycoproteins 640:652 glycoproteins 640:652 glycoproteins 640:652 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 2 33 theme tags 530:533 arg1 installation 486:497 the selective installation 472:497 the selective installation of visualization or enrichment tags 472:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 5 34 theme donor 922:926 arg1 sugars 928:933 MCR donor sugars 918:933 MCR donor sugars 918:933 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 1 35 used used 285:288 arg2 labs 275:278 many different labs 260:278 many different labs 260:278 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 3 36 theme glycoproteins 640:652 arg1 identification 622:635 the unbiased identification 609:635 the unbiased identification of glycoproteins 609:652 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 4 37 theme most 773:776 arg1 MCRs 778:781 most MCRs 773:781 most MCRs 773:781 For example, they are incorporated substoichiometrically into glycans, and most MCRs are not selective for one class (e.g., O-GlcNAcylation) of glycoprotein. 28231413 2 38 theme enrichment 519:528 arg1 tags 530:533 enrichment tags 519:533 enrichment tags 519:533 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 3 39 theme metabolic 542:550 arg1 reporters 561:569 These metabolic chemical reporters 536:569 These metabolic chemical reporters (MCRs) 536:576 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 3 39 theme metabolic 542:550 arg1 MCRs 572:575 MCRs 572:575 MCRs 572:575 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 5 40 theme MCR 918:920 arg1 sugars 928:933 MCR donor sugars 918:933 MCR donor sugars 918:933 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 1 41 theme many 260:263 arg1 labs 275:278 many different labs 260:278 many different labs 260:278 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 3 42 theme certain 677:683 arg1 limitations 685:695 certain limitations 677:695 certain limitations 677:695 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 3 43 theme chemical 552:559 arg1 reporters 561:569 These metabolic chemical reporters 536:569 These metabolic chemical reporters (MCRs) 536:576 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 3 43 theme chemical 552:559 arg1 MCRs 572:575 MCRs 572:575 MCRs 572:575 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 1 44 theme different 265:273 arg1 labs 275:278 many different labs 260:278 many different labs 260:278 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 0 45 theme Glycoprotein 83:94 arg1 Labeling 96:103 Glycoprotein Labeling 83:103 Glycoprotein Labeling 83:103 Metabolic Chemical Reporters of Glycans Exhibit Cell-Type-Selective Metabolism and Glycoprotein Labeling. 28231413 6 46 theme donor 1088:1092 arg1 sugars 1094:1099 the different donor sugars 1074:1099 the different donor sugars 1074:1099 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 4 47 theme glycoprotein 842:853 arg1 O-GlcNAcylation 822:836 O-GlcNAcylation 822:836 O-GlcNAcylation 822:836 For example, they are incorporated substoichiometrically into glycans, and most MCRs are not selective for one class (e.g., O-GlcNAcylation) of glycoprotein. 28231413 4 47 theme glycoprotein 842:853 arg1 class 809:813 one class 805:813 one class (e.g., O-GlcNAcylation) of glycoprotein 805:853 For example, they are incorporated substoichiometrically into glycans, and most MCRs are not selective for one class (e.g., O-GlcNAcylation) of glycoprotein. 28231413 2 48 theme unnatural 405:413 arg1 monosaccharides 415:429 these unnatural monosaccharides 399:429 these unnatural monosaccharides 399:429 A subset of these unnatural monosaccharides contain bioorthogonal groups that enable the selective installation of visualization or enrichment tags. 28231413 6 49 theme overall 1126:1132 arg1 levels 1143:1148 the overall labeling levels 1122:1148 the overall labeling levels 1122:1148 We found that the buildup of the different donor sugars correlated well with the overall labeling levels but less so with intracellular labeling of proteins by O-GlcNAcylation. 28231413 1 50 theme living 373:378 arg1 cells 380:384 living cells 373:384 living cells 373:384 Since the pioneering work by Reutter and co-workers that demonstrated structural flexibility in the carbohydrate biosynthesis and glycosylation pathways, many different labs have used unnatural monosaccharide analogues to perform glycan engineering on the surface of living cells. 28231413 5 51 theme sugars 928:933 arg1 biosynthesis 902:913 the biosynthesis 898:913 the biosynthesis of MCR donor sugars in cells 898:942 Here, we explore the relationship between the biosynthesis of MCR donor sugars in cells and the labeling levels of four different N-acetylglucosamine- and N-acetylgalactosamine-based MCRs. 28231413 3 52 contain have 672:675 arg2 limitations 685:695 certain limitations 677:695 certain limitations 677:695 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 28231413 3 52 contain have 672:675 arg1 they 664:667 they 664:667 they 664:667 These metabolic chemical reporters (MCRs) have proven to be powerful for the unbiased identification of glycoproteins; however, they do have certain limitations. 27523326 4 0 theme homologous 680:689 arg1 proteins 691:698 90% homologous proteins 676:698 90% homologous proteins 676:698 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 5 1 theme amino 760:764 arg1 patterns 771:778 amino acid patterns 760:778 amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated") 760:860 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 6 2 theme new 893:895 arg1 data 897:900 new data 893:900 new data 893:900 GlycoSiteAlign will integrate new data as they become available to confirm and expand results. 27523326 1 3 theme glycosylation 158:170 arg1 sites 172:176 glycosylation sites 158:176 glycosylation sites 158:176 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 5 4 dep features 806:813 arg1 " 834:834 "fucosylated" 822:834 "fucosylated" 822:834 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 5 4 dep features 806:813 arg1 " 859:859 "non-fucosylated" 843:859 "non-fucosylated" 843:859 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 2 5 theme characteristic 291:304 arg1 patterns 317:324 characteristic amino acid patterns 291:324 characteristic amino acid patterns of unique glycan-protein interactions 291:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 5 6 gly non-fucosylated 844:858 arg1 " 859:859 "non-fucosylated" 843:859 "non-fucosylated" 843:859 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 3 7 theme Bioinformatics 485:498 arg1 portal 509:514 Bioinformatics resource portal 485:514 Bioinformatics resource portal 485:514 GlycoSiteAlign uses data from the UniCarbKB and UniProtKB databases, and it is hosted on ExPASy, the Swiss Institute of Bioinformatics resource portal. 27523326 5 8 theme downloads 723:731 arg1 alignments 733:742 downloads alignments 723:742 downloads alignments 723:742 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 1 9 theme amino 106:110 arg1 sequences 117:125 amino acid sequences 106:125 amino acid sequences of variable length surrounding glycosylation sites 106:176 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 7 10 theme particular 1102:1111 arg1 glycosite 1113:1121 a particular glycosite 1100:1121 a particular glycosite 1100:1121 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 3 11 theme UniProtKB 413:421 arg1 databases 423:431 the UniCarbKB and UniProtKB databases 395:431 the UniCarbKB and UniProtKB databases 395:431 GlycoSiteAlign uses data from the UniCarbKB and UniProtKB databases, and it is hosted on ExPASy, the Swiss Institute of Bioinformatics resource portal. 27523326 5 12 theme selected 797:804 arg1 features 806:813 selected features 797:813 selected features (e.g., "fucosylated" versus "non-fucosylated") 797:860 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 5 13 theme non-fucosylated 844:858 arg1 " 859:859 "non-fucosylated" 843:859 "non-fucosylated" 843:859 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 0 14 theme GlycoSiteAlign 0:13 arg1 Alignment 26:34 GlycoSiteAlign: Glycosite Alignment 0:34 GlycoSiteAlign: Glycosite Alignment 0:34 GlycoSiteAlign: Glycosite Alignment Based on Glycan Structure. 27523326 4 15 theme glycan 564:569 arg1 features 571:578 either specific or general glycan features 537:578 either specific or general glycan features 537:578 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 5 16 gly fucosylated 823:833 arg1 " 834:834 "fucosylated" 822:834 "fucosylated" 822:834 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 2 17 theme patterns 317:324 arg1 identification 273:286 the identification 269:286 the identification of characteristic amino acid patterns of unique glycan-protein interactions 269:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 4 18 theme general 556:562 arg1 features 571:578 either specific or general glycan features 537:578 either specific or general glycan features 537:578 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 7 19 theme glycan 1080:1085 arg1 structure 1087:1095 a particular glycan structure 1067:1095 a particular glycan structure 1067:1095 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 2 20 theme acid 312:315 arg1 patterns 317:324 characteristic amino acid patterns 291:324 characteristic amino acid patterns of unique glycan-protein interactions 291:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 3 21 theme UniCarbKB 399:407 arg1 databases 423:431 the UniCarbKB and UniProtKB databases 395:431 the UniCarbKB and UniProtKB databases 395:431 GlycoSiteAlign uses data from the UniCarbKB and UniProtKB databases, and it is hosted on ExPASy, the Swiss Institute of Bioinformatics resource portal. 27523326 2 22 theme amino 306:310 arg1 patterns 317:324 characteristic amino acid patterns 291:324 characteristic amino acid patterns of unique glycan-protein interactions 291:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 5 23 theme acid 766:769 arg1 patterns 771:778 amino acid patterns 760:778 amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated") 760:860 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 2 24 theme interactions 351:362 arg1 patterns 317:324 characteristic amino acid patterns 291:324 characteristic amino acid patterns of unique glycan-protein interactions 291:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 5 25 theme fucosylated 823:833 arg1 " 834:834 "fucosylated" 822:834 "fucosylated" 822:834 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 1 26 theme glycan 208:213 arg1 structure 215:223 glycan structure 208:223 glycan structure 208:223 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 0 27 theme Glycosite 16:24 arg1 Alignment 26:34 GlycoSiteAlign: Glycosite Alignment 0:34 GlycoSiteAlign: Glycosite Alignment 0:34 GlycoSiteAlign: Glycosite Alignment Based on Glycan Structure. 27523326 5 28 theme tool 705:708 arg1 previews 710:717 tool previews 705:717 tool previews 705:717 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 7 29 gly glycosite 1113:1121 arg2 glycosite 1113:1121 a particular glycosite 1100:1121 a particular glycosite 1100:1121 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 7 30 theme promising 979:987 arg1 It 958:959 It 958:959 It 958:959 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 7 30 theme promising 979:987 arg1 tool 989:992 a promising tool 977:992 a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools 977:1197 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 1 31 theme structure 215:223 arg1 knowledge 195:203 the knowledge 191:203 the knowledge of glycan structure 191:223 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 5 32 dep previews 710:717 arg1 The 701:703 The 701:703 The 701:703 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 3 33 theme resource 500:507 arg1 portal 509:514 Bioinformatics resource portal 485:514 Bioinformatics resource portal 485:514 GlycoSiteAlign uses data from the UniCarbKB and UniProtKB databases, and it is hosted on ExPASy, the Swiss Institute of Bioinformatics resource portal. 27523326 0 34 theme Glycan 45:50 arg1 Structure 52:60 Glycan Structure 45:60 Glycan Structure 45:60 GlycoSiteAlign: Glycosite Alignment Based on Glycan Structure. 27523326 4 35 with alignment 637:645 arg1 option 656:661 the option 652:661 the option of including 90% homologous proteins 652:698 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 1 36 theme acid 112:115 arg1 sequences 117:125 amino acid sequences 106:125 amino acid sequences of variable length surrounding glycosylation sites 106:176 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 4 37 theme specific 544:551 arg1 features 571:578 either specific or general glycan features 537:578 either specific or general glycan features 537:578 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 1 38 gly glycosylation 158:170 arg2 sites 172:176 glycosylation sites 158:176 glycosylation sites 158:176 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 2 39 theme glycan-protein 336:349 arg1 interactions 351:362 unique glycan-protein interactions 329:362 unique glycan-protein interactions 329:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 7 40 attach link 1062:1065 arg1 glycosite 1113:1121 a particular glycosite 1100:1121 a particular glycosite 1100:1121 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 7 40 attach link 1062:1065 arg2 constraints 1045:1055 the constraints 1041:1055 the constraints that link a particular glycan structure to a particular glycosite 1041:1121 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 4 41 theme protein 603:609 arg1 fragments 611:619 the protein fragments 599:619 the protein fragments 599:619 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 27523326 5 42 dep " 834:834 arg1 e.g. 816:819 e.g. 816:819 e.g. 816:819 The tool previews and downloads alignments that may reveal amino acid patterns corresponding to selected features (e.g., "fucosylated" versus "non-fucosylated"). 27523326 7 43 theme particular 1069:1078 arg1 structure 1087:1095 a particular glycan structure 1067:1095 a particular glycan structure 1067:1095 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 2 44 theme unique 329:334 arg1 interactions 351:362 unique glycan-protein interactions 329:362 unique glycan-protein interactions 329:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 1 45 theme variable 130:137 arg1 length 139:144 variable length 130:144 variable length 130:144 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 7 46 theme prediction 1182:1191 arg1 tools 1193:1197 prediction tools 1182:1197 prediction tools 1182:1197 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 3 47 from databases 423:431 arg1 data 385:388 data 385:388 data from the UniCarbKB and UniProtKB databases 385:431 GlycoSiteAlign uses data from the UniCarbKB and UniProtKB databases, and it is hosted on ExPASy, the Swiss Institute of Bioinformatics resource portal. 27523326 2 48 theme exploratory 235:245 arg1 resource 247:254 an exploratory resource 232:254 an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions 232:362 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 2 48 theme exploratory 235:245 arg1 It 226:227 It 226:227 It 226:227 It is an exploratory resource intended for the identification of characteristic amino acid patterns of unique glycan-protein interactions. 27523326 1 49 theme length 139:144 arg1 sequences 117:125 amino acid sequences 106:125 amino acid sequences of variable length surrounding glycosylation sites 106:176 GlycoSiteAlign is a tool designed to align amino acid sequences of variable length surrounding glycosylation sites depending on the knowledge of glycan structure. 27523326 7 50 theme long 1135:1138 arg1 term 1140:1143 the long term 1131:1143 the long term 1131:1143 It is presented as a promising tool for assessing and refining the knowledge about the constraints that link a particular glycan structure to a particular glycosite, and in the long term, this application could help improve prediction tools. 27523326 4 51 theme fragments 611:619 arg1 length 589:594 the length 585:594 the length of the protein fragments 585:619 The user can select either specific or general glycan features, set the length of the protein fragments, and trigger an alignment with the option of including 90% homologous proteins. 25004930 0 0 theme mass 62:65 arg1 method 107:112 a new mass spectrometric high-throughput profiling method 56:112 a new mass spectrometric high-throughput profiling method 56:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 10 1 theme antigen 1841:1847 arg1 binding 1849:1855 antigen binding 1841:1855 antigen binding 1841:1855 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 8 2 theme glycans 1424:1430 arg1 galactosylation 1401:1415 nearly complete galactosylation 1385:1415 nearly complete galactosylation of Fab glycans 1385:1430 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 4 3 theme reflectron 669:678 arg1 mode 689:692 reflectron positive mode 669:692 reflectron positive mode 669:692 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 10 4 from roles 1727:1731 arg1 immunity 1759:1766 (auto)immunity 1753:1766 (auto)immunity 1753:1766 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 0 5 theme new 58:60 arg1 method 107:112 a new mass spectrometric high-throughput profiling method 56:112 a new mass spectrometric high-throughput profiling method 56:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 6 theme Data 887:890 arg1 analysis 892:899 Data analysis 887:899 Data analysis 887:899 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 10 7 theme glycans 1742:1748 arg1 roles 1727:1731 roles 1727:1731 roles 1727:1731 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 8 theme Fc 1665:1666 arg1 set 1672:1674 set 1672:1674 set up in this paper 1672:1691 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 8 theme Fc 1665:1666 arg1 analysis 1641:1648 the parallel analysis 1628:1648 the parallel analysis of IgG Fab and Fc 1628:1666 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 8 theme Fc 1665:1666 arg1 important 1702:1710 important 1702:1710 important 1702:1710 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 3 9 gly glycosylation 512:524 arg1 IgG 540:542 polyclonal IgG 529:542 polyclonal IgG purified from 5 μl of serum 529:570 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 4 10 from analysis 657:664 arg1 mode 689:692 reflectron positive mode 669:692 reflectron positive mode 669:692 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 10 11 dep important 1702:1710 arg1 important 1702:1710 important 1702:1710 important 1702:1710 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 11 dep important 1702:1710 arg1 set 1672:1674 set 1672:1674 set up in this paper 1672:1691 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 11 dep important 1702:1710 arg1 analysis 1641:1648 the parallel analysis 1628:1648 the parallel analysis of IgG Fab and Fc 1628:1666 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 0 12 theme high-throughput 81:95 arg1 method 107:112 a new mass spectrometric high-throughput profiling method 56:112 a new mass spectrometric high-throughput profiling method 56:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 3 13 theme polyclonal 529:538 arg1 IgG 540:542 polyclonal IgG 529:542 polyclonal IgG purified from 5 μl of serum 529:570 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 10 14 theme IgG 1653:1655 arg1 Fab 1657:1659 IgG Fab 1653:1659 IgG Fab 1653:1659 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 0 15 theme spectrometric 67:79 arg1 method 107:112 a new mass spectrometric high-throughput profiling method 56:112 a new mass spectrometric high-throughput profiling method 56:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 10 16 theme auto 1754:1757 arg1 immunity 1759:1766 (auto)immunity 1753:1766 (auto)immunity 1753:1766 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 4 17 theme sialic 743:748 arg1 acid 750:753 sialic acid 743:753 sialic acid 743:753 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 3 18 theme IgG 540:542 arg1 Fab 501:503 Fab 501:503 Fab 501:503 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 3 18 theme IgG 540:542 arg1 glycosylation 512:524 Fc glycosylation 509:524 Fc glycosylation 509:524 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 1 19 theme pathological 261:272 arg1 events 292:297 pathological and physiological events 261:297 pathological and physiological events 261:297 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 5 20 theme cohort 810:815 arg1 samples 787:793 174 samples 783:793 174 samples of a pregnancy cohort 783:815 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 3 21 theme high-throughput 465:479 arg1 workflow 481:488 a high-throughput workflow 463:488 a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum 463:570 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 5 22 theme Fab 827:829 arg1 features 845:852 Fab glycosylation features 827:852 Fab glycosylation features 827:852 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 0 23 theme profiling 97:105 arg1 method 107:112 a new mass spectrometric high-throughput profiling method 56:112 a new mass spectrometric high-throughput profiling method 56:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 8 24 theme Fab 1278:1280 arg1 sialylation 1291:1301 Fab N-glycan sialylation 1278:1301 Fab N-glycan sialylation 1278:1301 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 5 25 theme 174 783:785 arg1 samples 787:793 174 samples 783:793 174 samples of a pregnancy cohort 783:815 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 2 26 theme IgG 442:444 arg1 IgG 442:444 IgG 442:444 IgG 442:444 In contrast, little is known about Fab-linked N-glycosylation, carried by ∼ 20% of IgG. 25004930 2 26 theme IgG 442:444 arg1 % 437:437 20% 435:437 20% of IgG 435:444 In contrast, little is known about Fab-linked N-glycosylation, carried by ∼ 20% of IgG. 25004930 1 27 theme constant 186:193 arg1 fragment 195:202 the constant fragment 182:202 the constant fragment (Fc) of immunoglobulin G 182:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 1 27 theme constant 186:193 arg1 Fc 205:206 Fc 205:206 Fc 205:206 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 0 28 theme G 15:15 arg1 analysis 41:48 Immunoglobulin G (IgG) Fab glycosylation analysis 0:48 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method 0:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 29 theme Fab 1125:1127 arg1 portion 1129:1135 the Fab portion 1121:1135 the Fab portion 1121:1135 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 4 30 theme linkage-specific 708:723 arg1 derivatization 725:738 a novel linkage-specific derivatization 700:738 a novel linkage-specific derivatization of sialic acid 700:753 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 7 31 theme several 1225:1231 arg1 changes 1233:1239 several changes 1225:1239 several changes 1225:1239 Additionally, we observed several changes during pregnancy and after delivery. 25004930 1 32 theme fragment 195:202 arg1 glycosylation 165:177 The N-linked glycosylation 152:177 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G 152:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 0 33 theme Immunoglobulin 0:13 arg1 IgG 18:20 IgG 18:20 IgG 18:20 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 0 33 theme Immunoglobulin 0:13 arg1 G 15:15 Immunoglobulin G 0:15 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method 0:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 34 theme marked 910:915 arg1 differences 917:927 marked differences 910:927 marked differences between Fab and Fc glycosylation 910:960 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 4 35 theme novel 702:706 arg1 derivatization 725:738 a novel linkage-specific derivatization 700:738 a novel linkage-specific derivatization of sialic acid 700:753 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 6 36 theme GlcNAc 1047:1052 arg1 levels 981:986 the levels 977:986 the levels of galactosylation and sialylation 977:1021 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 36 theme GlcNAc 1047:1052 arg1 incidence 1024:1032 incidence 1024:1032 incidence of bisecting GlcNAc 1024:1052 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 36 theme GlcNAc 1047:1052 arg1 presence 1059:1066 presence 1059:1066 presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation 1059:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 1 37 theme physiological 278:290 arg1 events 292:297 pathological and physiological events 261:297 pathological and physiological events 261:297 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 0 38 theme pregnancy-associated 122:141 arg1 changes 143:149 pregnancy-associated changes 122:149 pregnancy-associated changes 122:149 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 39 theme higher 1168:1173 arg1 levels 1175:1180 higher levels 1168:1180 higher levels of fucosylation 1168:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 8 40 theme time 1368:1371 arg1 points 1373:1378 postpartum time points 1357:1378 postpartum time points 1357:1378 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 5 41 theme pregnancy 800:808 arg1 cohort 810:815 a pregnancy cohort 798:815 a pregnancy cohort 798:815 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 6 42 theme Fc 945:946 arg1 glycosylation 948:960 Fc glycosylation 945:960 Fc glycosylation 945:960 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 8 43 theme Fab 1420:1422 arg1 glycans 1424:1430 Fab glycans 1420:1430 Fab glycans 1420:1430 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 8 44 theme N-glycan 1282:1289 arg1 sialylation 1291:1301 Fab N-glycan sialylation 1278:1301 Fab N-glycan sialylation 1278:1301 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 5 45 theme glycosylation 831:843 arg1 features 845:852 Fab glycosylation features 827:852 Fab glycosylation features 827:852 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 0 46 theme Fab 23:25 arg1 analysis 41:48 Immunoglobulin G (IgG) Fab glycosylation analysis 0:48 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method 0:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 47 theme fucosylation 1185:1196 arg1 levels 1175:1180 higher levels 1168:1180 higher levels of fucosylation 1168:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 10 48 theme parallel 1632:1639 arg1 set 1672:1674 set 1672:1674 set up in this paper 1672:1691 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 48 theme parallel 1632:1639 arg1 analysis 1641:1648 the parallel analysis 1628:1648 the parallel analysis of IgG Fab and Fc 1628:1666 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 48 theme parallel 1632:1639 arg1 important 1702:1710 important 1702:1710 important 1702:1710 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 4 49 theme different 612:620 arg1 N-glycans 622:630 37 different N-glycans 609:630 37 different N-glycans 609:630 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 6 50 theme sialylation 1011:1021 arg1 levels 981:986 the levels 977:986 the levels of galactosylation and sialylation 977:1021 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 50 theme sialylation 1011:1021 arg1 incidence 1024:1032 incidence 1024:1032 incidence of bisecting GlcNAc 1024:1052 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 50 theme sialylation 1011:1021 arg1 presence 1059:1066 presence 1059:1066 presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation 1059:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 1 51 link N-linked 156:163 arg1 glycosylation 165:177 The N-linked glycosylation 152:177 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G 152:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 8 52 theme postpartum 1357:1366 arg1 points 1373:1378 postpartum time points 1357:1378 postpartum time points 1357:1378 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 1 53 theme immunoglobulin 212:225 arg1 G 227:227 immunoglobulin G 212:227 immunoglobulin G 212:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 5 54 with features 845:852 arg1 pregnancy 876:884 pregnancy 876:884 pregnancy 876:884 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 2 55 link Fab-linked 394:403 arg1 N-glycosylation 405:419 Fab-linked N-glycosylation 394:419 Fab-linked N-glycosylation 394:419 In contrast, little is known about Fab-linked N-glycosylation, carried by ∼ 20% of IgG. 25004930 9 56 theme glycosylation 1460:1472 arg1 changes 1474:1480 Fc glycosylation changes 1457:1480 Fc glycosylation changes 1457:1480 Fc glycosylation changes were similar to results described before, with increased galactosylation and sialylation and decreased bisection during pregnancy. 25004930 0 57 theme glycosylation 27:39 arg1 analysis 41:48 Immunoglobulin G (IgG) Fab glycosylation analysis 0:48 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method 0:112 Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes. 25004930 6 58 theme high 1071:1074 arg1 structures 1084:1093 high mannose structures 1071:1093 high mannose structures 1071:1093 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 10 59 theme binding 1849:1855 arg1 modulation 1827:1836 modulation 1827:1836 modulation of antigen binding 1827:1855 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 59 theme binding 1849:1855 arg1 recognition 1795:1805 recognition 1795:1805 recognition by human lectins 1795:1822 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 3 60 theme serum 566:570 arg1 μl 560:561 5 μl 558:561 5 μl of serum 558:570 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 4 61 theme positive 680:687 arg1 mode 689:692 reflectron positive mode 669:692 reflectron positive mode 669:692 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 1 62 theme antibody 325:332 arg1 properties 347:356 antibody inflammatory properties 325:356 antibody inflammatory properties 325:356 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 4 63 theme MALDI-TOF-MS 644:655 arg1 analysis 657:664 MALDI-TOF-MS analysis 644:664 MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid 644:753 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 1 64 theme G 227:227 arg1 fragment 195:202 the constant fragment 182:202 the constant fragment (Fc) of immunoglobulin G 182:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 1 64 theme G 227:227 arg1 Fc 205:206 Fc 205:206 Fc 205:206 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 1 65 gly glycosylation 165:177 arg1 fragment 195:202 the constant fragment 182:202 the constant fragment (Fc) of immunoglobulin G 182:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 1 65 gly glycosylation 165:177 arg1 Fc 205:206 Fc 205:206 Fc 205:206 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 10 66 theme human 1810:1814 arg1 lectins 1816:1822 human lectins 1810:1822 human lectins 1810:1822 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 4 67 theme acid 750:753 arg1 derivatization 725:738 a novel linkage-specific derivatization 700:738 a novel linkage-specific derivatization of sialic acid 700:753 We were able to detect and quantify 37 different N-glycans by means of MALDI-TOF-MS analysis in reflectron positive mode using a novel linkage-specific derivatization of sialic acid. 25004930 10 68 theme Fab 1657:1659 arg1 set 1672:1674 set 1672:1674 set up in this paper 1672:1691 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 68 theme Fab 1657:1659 arg1 analysis 1641:1648 the parallel analysis 1628:1648 the parallel analysis of IgG Fab and Fc 1628:1666 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 10 68 theme Fab 1657:1659 arg1 important 1702:1710 important 1702:1710 important 1702:1710 We expect that the parallel analysis of IgG Fab and Fc, as set up in this paper, will be important for unraveling roles of these glycans in (auto)immunity, which may be mediated via recognition by human lectins or modulation of antigen binding. 25004930 3 69 theme Fc 509:510 arg1 glycosylation 512:524 Fc glycosylation 509:524 Fc glycosylation 509:524 Here we present a high-throughput workflow to analyze Fab and Fc glycosylation of polyclonal IgG purified from 5 μl of serum. 25004930 9 70 theme Fc 1457:1458 arg1 changes 1474:1480 Fc glycosylation changes 1457:1480 Fc glycosylation changes 1457:1480 Fc glycosylation changes were similar to results described before, with increased galactosylation and sialylation and decreased bisection during pregnancy. 25004930 6 71 theme mannose 1076:1082 arg1 structures 1084:1093 high mannose structures 1071:1093 high mannose structures 1071:1093 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 5 72 with change 864:869 arg1 pregnancy 876:884 pregnancy 876:884 pregnancy 876:884 This method was applied to 174 samples of a pregnancy cohort to reveal Fab glycosylation features and their change with pregnancy. 25004930 6 73 theme bisecting 1037:1045 arg1 GlcNAc 1047:1052 bisecting GlcNAc 1037:1052 bisecting GlcNAc 1037:1052 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 74 theme structures 1084:1093 arg1 levels 981:986 the levels 977:986 the levels of galactosylation and sialylation 977:1021 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 74 theme structures 1084:1093 arg1 incidence 1024:1032 incidence 1024:1032 incidence of bisecting GlcNAc 1024:1052 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 74 theme structures 1084:1093 arg1 presence 1059:1066 presence 1059:1066 presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation 1059:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 2 75 theme Fab-linked 394:403 arg1 N-glycosylation 405:419 Fab-linked N-glycosylation 394:419 Fab-linked N-glycosylation 394:419 In contrast, little is known about Fab-linked N-glycosylation, carried by ∼ 20% of IgG. 25004930 6 76 theme galactosylation 991:1005 arg1 levels 981:986 the levels 977:986 the levels of galactosylation and sialylation 977:1021 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 76 theme galactosylation 991:1005 arg1 incidence 1024:1032 incidence 1024:1032 incidence of bisecting GlcNAc 1024:1052 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 6 76 theme galactosylation 991:1005 arg1 presence 1059:1066 presence 1059:1066 presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation 1059:1196 Data analysis revealed marked differences between Fab and Fc glycosylation, especially in the levels of galactosylation and sialylation, incidence of bisecting GlcNAc, and presence of high mannose structures, which were all higher in the Fab portion than the Fc, whereas Fc showed higher levels of fucosylation. 25004930 1 77 theme inflammatory 334:345 arg1 properties 347:356 antibody inflammatory properties 325:356 antibody inflammatory properties 325:356 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25004930 8 78 theme complete 1392:1399 arg1 galactosylation 1401:1415 nearly complete galactosylation 1385:1415 nearly complete galactosylation of Fab glycans 1385:1430 Fab N-glycan sialylation was increased and bisection was decreased relative to postpartum time points, and nearly complete galactosylation of Fab glycans was observed throughout. 25004930 1 79 theme N-linked 156:163 arg1 glycosylation 165:177 The N-linked glycosylation 152:177 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G 152:227 The N-linked glycosylation of the constant fragment (Fc) of immunoglobulin G has been shown to change during pathological and physiological events and to strongly influence antibody inflammatory properties. 25205566 0 0 theme mass 91:94 arg1 spectrometry 96:107 chemical immobilization and mass spectrometry 63:107 spectrometry 96:107 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 3 1 theme Mass 344:347 arg1 spectrometry 349:360 Mass spectrometry 344:360 Mass spectrometry 344:360 Mass spectrometry has emerged as a major method for glycomic analysis. 25205566 1 2 theme modifications 162:174 arg1 modifications 162:174 most common protein modifications 142:174 most common protein modifications 142:174 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 1 2 theme modifications 162:174 arg1 one 135:137 one 135:137 one 135:137 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 6 3 theme solid 941:945 arg1 phase 947:951 solid phase 941:951 solid phase 941:951 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 4 4 theme mass 541:544 arg1 spectrometry 546:557 mass spectrometry 541:557 mass spectrometry 541:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 3 5 theme major 379:383 arg1 method 385:390 a major method 377:390 a major method for glycomic analysis 377:412 Mass spectrometry has emerged as a major method for glycomic analysis. 25205566 6 6 attach linked 849:854 arg3 support 886:892 the solid support 876:892 the solid support 876:892 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 5 7 theme chemoenzymatic 694:707 arg1 approach 709:716 a chemoenzymatic approach 692:716 a chemoenzymatic approach on solid-phase 692:731 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 6 8 theme glycan 957:962 arg1 derivatization 964:977 glycan derivatization 957:977 glycan derivatization 957:977 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 1 9 gly glycosylation 118:130 arg1 activities 211:220 many biological activities 195:220 many biological activities 195:220 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 4 10 theme glycan 424:429 arg1 steps 497:501 two critical steps 484:501 two critical steps in glycomic analysis of glycans using mass spectrometry 484:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 4 10 theme glycan 424:429 arg1 extraction 431:440 glycan extraction 424:440 glycan extraction from proteins and glycan modification 424:478 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 0 11 attach released 29:36 arg1 glycoproteins 43:55 glycoproteins 43:55 glycoproteins 43:55 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 0 11 attach released 29:36 arg2 glycans 21:27 glycans 21:27 glycans released from glycoproteins 21:55 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 6 12 dep glycans 828:834 arg1 linked 849:854 linked 849:854 linked to glycoproteins on the solid support 849:892 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 5 13 gly glycoproteins 672:684 arg1 glycoproteins 672:684 glycoproteins 672:684 glycoproteins 672:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 4 14 theme glycomic 506:513 arg1 analysis 515:522 glycomic analysis 506:522 glycomic analysis of glycans using mass spectrometry 506:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 5 15 theme glycans 659:665 arg1 modification 643:654 modification 643:654 modification 643:654 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 5 15 theme glycans 659:665 arg1 isolation 629:637 isolation 629:637 isolation 629:637 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 5 16 theme novel 592:596 arg1 method 618:623 a novel and high-throughput method 590:623 a novel and high-throughput method for isolation and modification of glycans from glycoproteins 590:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 6 17 gly glycoproteins 859:871 arg1 glycoproteins 859:871 glycoproteins 859:871 glycoproteins 859:871 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 0 18 theme Glycomic 0:7 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of glycans released from glycoproteins 0:55 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 4 19 from steps 497:501 arg1 analysis 515:522 glycomic analysis 506:522 glycomic analysis of glycans using mass spectrometry 506:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 1 20 theme many 195:198 arg1 activities 211:220 many biological activities 195:220 many biological activities 195:220 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 0 21 theme glycans 21:27 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of glycans released from glycoproteins 0:55 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 5 22 theme high-throughput 602:616 arg1 method 618:623 a novel and high-throughput method 590:623 a novel and high-throughput method for isolation and modification of glycans from glycoproteins 590:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 1 23 theme Protein 110:116 arg1 glycosylation 118:130 Protein glycosylation 110:130 Protein glycosylation 110:130 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 1 24 theme biological 200:209 arg1 activities 211:220 many biological activities 195:220 many biological activities 195:220 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 3 25 theme glycomic 396:403 arg1 analysis 405:412 glycomic analysis 396:412 glycomic analysis 396:412 Mass spectrometry has emerged as a major method for glycomic analysis. 25205566 5 26 from modification 643:654 arg1 glycoproteins 672:684 glycoproteins 672:684 glycoproteins 672:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 5 27 from isolation 629:637 arg1 glycoproteins 672:684 glycoproteins 672:684 glycoproteins 672:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 7 28 attach released 997:1004 arg1 support 1021:1027 the solid support 1011:1027 the solid support for analysis by mass spectrometry 1011:1061 Glycans are then released from the solid support for analysis by mass spectrometry. 25205566 7 28 attach released 997:1004 arg2 Glycans 980:986 Glycans 980:986 Glycans 980:986 Glycans are then released from the solid support for analysis by mass spectrometry. 25205566 8 29 theme glycomic 1130:1137 arg1 analysis 1139:1146 high-throughput glycomic analysis 1114:1146 high-throughput glycomic analysis from complex biological or clinical samples 1114:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 8 30 from samples 1184:1190 arg1 analysis 1139:1146 high-throughput glycomic analysis 1114:1146 high-throughput glycomic analysis from complex biological or clinical samples 1114:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 2 31 link N-linked 223:230 arg1 glycosylation 245:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 8 32 theme biological 1161:1170 arg1 samples 1184:1190 complex biological or clinical samples 1153:1190 complex biological or clinical samples 1153:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 4 33 from modification 467:478 arg1 steps 497:501 two critical steps 484:501 two critical steps in glycomic analysis of glycans using mass spectrometry 484:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 4 33 from modification 467:478 arg1 extraction 431:440 glycan extraction 424:440 glycan extraction from proteins and glycan modification 424:478 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 4 34 theme critical 488:495 arg1 steps 497:501 two critical steps 484:501 two critical steps in glycomic analysis of glycans using mass spectrometry 484:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 4 34 theme critical 488:495 arg1 extraction 431:440 glycan extraction 424:440 glycan extraction from proteins and glycan modification 424:478 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 6 35 theme solid 880:884 arg1 support 886:892 the solid support 876:892 the solid support 876:892 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 2 36 theme O-linked 236:243 arg1 glycosylation 245:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 7 37 theme mass 1045:1048 arg1 spectrometry 1050:1061 mass spectrometry 1045:1061 mass spectrometry 1045:1061 Glycans are then released from the solid support for analysis by mass spectrometry. 25205566 8 38 theme high-throughput 1114:1128 arg1 analysis 1139:1146 high-throughput glycomic analysis 1114:1146 high-throughput glycomic analysis from complex biological or clinical samples 1114:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 5 39 from glycoproteins 672:684 arg1 glycans 659:665 glycans 659:665 glycans from glycoproteins 659:684 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 5 39 from glycoproteins 672:684 arg1 modification 643:654 modification 643:654 modification 643:654 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 5 39 from glycoproteins 672:684 arg1 isolation 629:637 isolation 629:637 isolation 629:637 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 2 40 theme N-linked 223:230 arg1 glycosylation 245:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 6 41 theme unconjugated 788:799 arg1 molecules 801:809 unconjugated molecules 788:809 unconjugated molecules 788:809 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 4 42 from proteins 447:454 arg1 steps 497:501 two critical steps 484:501 two critical steps in glycomic analysis of glycans using mass spectrometry 484:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 4 42 from proteins 447:454 arg1 extraction 431:440 glycan extraction 424:440 glycan extraction from proteins and glycan modification 424:478 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 2 43 theme glycan 287:292 arg1 modifications 294:306 abundant glycan modifications 278:306 abundant glycan modifications 278:306 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 2 44 link O-linked 236:243 arg1 glycosylation 245:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation 223:257 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 8 45 theme complex 1153:1159 arg1 samples 1184:1190 complex biological or clinical samples 1153:1190 complex biological or clinical samples 1153:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 4 46 theme glycan 460:465 arg1 modification 467:478 glycan modification 460:478 glycan modification 460:478 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 2 47 theme abundant 278:285 arg1 modifications 294:306 abundant glycan modifications 278:306 abundant glycan modifications 278:306 N-linked and O-linked glycosylation not only represent abundant glycan modifications, but also are structurally diverse. 25205566 7 48 theme solid 1015:1019 arg1 support 1021:1027 the solid support 1011:1027 the solid support for analysis by mass spectrometry 1011:1061 Glycans are then released from the solid support for analysis by mass spectrometry. 25205566 8 49 theme clinical 1175:1182 arg1 samples 1184:1190 complex biological or clinical samples 1153:1190 complex biological or clinical samples 1153:1190 The procedures outlined are robust and useful for high-throughput glycomic analysis from complex biological or clinical samples. 25205566 0 50 gly glycoproteins 43:55 arg1 glycoproteins 43:55 glycoproteins 43:55 glycoproteins 43:55 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 6 51 theme solid 770:774 arg1 support 776:782 a solid support 768:782 a solid support 768:782 Proteins are first immobilized to a solid support and unconjugated molecules are washed away; glycans, while still linked to glycoproteins on the solid support, can be treated enzymatically or chemically on solid phase for glycan derivatization. 25205566 1 52 theme common 147:152 arg1 modifications 162:174 most common protein modifications 142:174 most common protein modifications 142:174 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 25205566 5 53 from approach 709:716 arg1 solid-phase 721:731 solid-phase 721:731 solid-phase 721:731 In this protocol, we describe a novel and high-throughput method for isolation and modification of glycans from glycoproteins using a chemoenzymatic approach on solid-phase. 25205566 0 54 theme chemical 63:70 arg1 immobilization 72:85 chemical immobilization and mass spectrometry 63:107 immobilization 72:85 Glycomic analysis of glycans released from glycoproteins using chemical immobilization and mass spectrometry. 25205566 4 55 theme glycans 527:533 arg1 analysis 515:522 glycomic analysis 506:522 glycomic analysis of glycans using mass spectrometry 506:557 However, glycan extraction from proteins and glycan modification are two critical steps in glycomic analysis of glycans using mass spectrometry. 25205566 1 56 theme protein 154:160 arg1 modifications 162:174 most common protein modifications 142:174 most common protein modifications 142:174 Protein glycosylation is one of most common protein modifications and is involved in many biological activities. 24692304 6 0 theme electron 906:913 arg1 map 923:925 the electron density map 902:925 the electron density map 902:925 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 3 1 theme glycan 505:510 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 4 2 theme quasi-racemic 591:603 arg1 mixture 605:611 a quasi-racemic mixture 589:611 a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein 589:679 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 0 3 theme forms 70:74 arg1 structures 22:31 (Quasi-)racemic X-ray structures 0:31 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 0:138 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 2 4 theme moiety 333:338 arg1 heterogeneity 287:299 the heterogeneity 283:299 the heterogeneity of the oligosaccharide (glycan) moiety 283:338 Glycoproteins can be hard to crystallize because of the heterogeneity of the oligosaccharide (glycan) moiety. 24692304 7 5 dep enantiomers 961:971 arg1 enantiomers 961:971 the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 949:997 the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 949:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 7 5 dep enantiomers 961:971 arg1 D-Ser-CCL1 988:997 D-Ser-CCL1 988:997 D-Ser-CCL1 988:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 7 5 dep enantiomers 961:971 arg1 L-Ser-CCL1 973:982 L-Ser-CCL1 973:982 L-Ser-CCL1 973:982 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 0 6 theme chemokine 83:91 arg1 Ser-CCL1 93:100 the chemokine Ser-CCL1 79:100 the chemokine Ser-CCL1 prepared by total chemical synthesis 79:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 1 7 gly glycosylated 198:209 arg1 Ser-CCL1 221:228 the glycosylated chemokine Ser-CCL1 194:228 the glycosylated chemokine Ser-CCL1 194:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 8 8 theme glycosylated-L-Ser-CCL1 1179:1201 arg1 moieties 1152:1159 the protein moieties 1140:1159 the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 1140:1201 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 0 9 gly non-glycosylated 53:68 arg1 forms 70:74 glycosylated and non-glycosylated forms 36:74 glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis 36:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 0 9 gly non-glycosylated 53:68 arg1 Ser-CCL1 93:100 the chemokine Ser-CCL1 79:100 the chemokine Ser-CCL1 prepared by total chemical synthesis 79:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 3 10 theme asialo 471:476 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 8 11 theme moieties 1152:1159 arg1 Superimposition 1103:1117 Superimposition 1103:1117 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 1103:1201 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 3 12 theme covalent 530:537 arg1 structure 539:547 defined covalent structure 522:547 defined covalent structure 522:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 0 13 theme Ser-CCL1 93:100 arg1 forms 70:74 glycosylated and non-glycosylated forms 36:74 glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis 36:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 6 14 theme density 915:921 arg1 map 923:925 the electron density map 902:925 the electron density map 902:925 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 7 15 theme 2.7-2.15 Å 1091:1100 arg1 resolution 1077:1086 a resolution 1075:1086 a resolution of 2.7-2.15 Å 1075:1100 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 3 16 theme structure 539:547 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 0 17 theme Quasi- 1:6 arg1 structures 22:31 (Quasi-)racemic X-ray structures 0:31 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 0:138 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 2 18 gly heterogeneity 287:299 arg1 moiety 333:338 the oligosaccharide (glycan) moiety 304:338 the oligosaccharide (glycan) moiety 304:338 Glycoproteins can be hard to crystallize because of the heterogeneity of the oligosaccharide (glycan) moiety. 24692304 3 19 used used 344:347 arg2 We 341:342 We 341:342 We 341:342 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 6 20 theme glycan 821:826 arg1 moiety 828:833 the glycan moiety 817:833 the glycan moiety 817:833 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 8 21 theme structures 1126:1135 arg1 Superimposition 1103:1117 Superimposition 1103:1117 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 1103:1201 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 3 22 theme chemical 403:410 arg1 synthesis 412:420 total chemical synthesis 397:420 total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 397:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 3 23 gly glycosylated 349:360 arg1 Ser-CCL1 362:369 glycosylated Ser-CCL1 349:369 glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 349:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 6 24 from well-defined 886:897 arg1 map 923:925 the electron density map 902:925 the electron density map 902:925 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 1 25 theme X-ray 167:171 arg1 structure 181:189 the X-ray crystal structure 163:189 the X-ray crystal structure of the glycosylated chemokine Ser-CCL1 163:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 0 26 theme racemic 8:14 arg1 structures 22:31 (Quasi-)racemic X-ray structures 0:31 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 0:138 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 4 27 theme crystal 557:563 arg1 formation 565:573 Facile crystal formation 550:573 Facile crystal formation 550:573 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 0 28 theme total 114:118 arg1 synthesis 129:137 total chemical synthesis 114:137 total chemical synthesis 114:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 7 29 theme protein 953:959 arg1 enantiomers 961:971 the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 949:997 the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 949:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 7 29 theme protein 953:959 arg1 D-Ser-CCL1 988:997 D-Ser-CCL1 988:997 D-Ser-CCL1 988:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 7 29 theme protein 953:959 arg1 L-Ser-CCL1 973:982 L-Ser-CCL1 973:982 L-Ser-CCL1 973:982 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 1 30 theme crystal 173:179 arg1 structure 181:189 the X-ray crystal structure 163:189 the X-ray crystal structure of the glycosylated chemokine Ser-CCL1 163:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 4 31 theme glycosylated 627:638 arg1 L-protein 640:648 L-protein 640:648 L-protein 640:648 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 6 32 link N-linked 860:867 arg1 well-defined 886:897 well-defined 886:897 well-defined 886:897 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 6 32 link N-linked 860:867 arg1 sugar 876:880 only the N-linked GlcNAc sugar 851:880 only the N-linked GlcNAc sugar 851:880 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 8 33 theme protein 1255:1261 arg1 structure 1263:1271 the protein structure 1251:1271 the protein structure 1251:1271 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 3 34 theme nonasaccharide 490:503 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 3 35 theme homogeneous 427:437 arg1 compound 439:446 a homogeneous compound 425:446 a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 425:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 0 36 theme X-ray 16:20 arg1 structures 22:31 (Quasi-)racemic X-ray structures 0:31 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 0:138 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 0 37 theme chemical 120:127 arg1 synthesis 129:137 total chemical synthesis 114:137 total chemical synthesis 114:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 8 38 theme L-Ser-CCL1 1164:1173 arg1 moieties 1152:1159 the protein moieties 1140:1159 the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 1140:1201 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 3 39 theme glycosylated 349:360 arg1 Ser-CCL1 362:369 glycosylated Ser-CCL1 349:369 glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 349:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 6 40 from map 923:925 arg1 well-defined 886:897 well-defined 886:897 well-defined 886:897 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 6 40 from map 923:925 arg1 sugar 876:880 only the N-linked GlcNAc sugar 851:880 only the N-linked GlcNAc sugar 851:880 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 0 41 theme glycosylated 36:47 arg1 forms 70:74 glycosylated and non-glycosylated forms 36:74 glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis 36:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 3 42 link N-linked 462:469 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 3 43 contain containing 448:457 arg1 compound 439:446 a homogeneous compound 425:446 a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 425:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 3 43 contain containing 448:457 arg2 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 7 44 theme true 1047:1050 arg1 racemate 1052:1059 the true racemate 1043:1059 the true racemate 1043:1059 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 3 45 theme biantennary 478:488 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 8 46 theme structure 1263:1271 arg1 alteration 1237:1246 no significant alteration 1222:1246 no significant alteration of the protein structure by N-glycosylation 1222:1290 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 4 47 theme Facile 550:555 arg1 formation 565:573 Facile crystal formation 550:573 Facile crystal formation 550:573 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 4 48 gly glycosylated 723:734 arg1 L-protein 736:744 the glycosylated L-protein 719:744 the glycosylated L-protein alone 719:750 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 7 49 theme enantiomers 961:971 arg1 mixture 938:944 A racemic mixture 928:944 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 928:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 2 50 theme glycan 325:330 arg1 moiety 333:338 the oligosaccharide (glycan) moiety 304:338 the oligosaccharide (glycan) moiety 304:338 Glycoproteins can be hard to crystallize because of the heterogeneity of the oligosaccharide (glycan) moiety. 24692304 8 51 theme significant 1225:1235 arg1 alteration 1237:1246 no significant alteration 1222:1246 no significant alteration of the protein structure by N-glycosylation 1222:1290 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 7 52 theme racemate 1052:1059 arg1 structure 1030:1038 the structure 1026:1038 the structure of the true racemate 1026:1059 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 6 53 theme GlcNAc 869:874 arg1 well-defined 886:897 well-defined 886:897 well-defined 886:897 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 6 53 theme GlcNAc 869:874 arg1 sugar 876:880 only the N-linked GlcNAc sugar 851:880 only the N-linked GlcNAc sugar 851:880 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 4 54 gly glycosylated 627:638 arg1 L-protein 640:648 L-protein 640:648 L-protein 640:648 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 3 55 theme N-linked 462:469 arg1 moiety 512:517 an N-linked asialo biantennary nonasaccharide glycan moiety 459:517 an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 459:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 1 56 theme glycosylated 198:209 arg1 Ser-CCL1 221:228 the glycosylated chemokine Ser-CCL1 194:228 the glycosylated chemokine Ser-CCL1 194:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 6 57 theme N-linked 860:867 arg1 well-defined 886:897 well-defined 886:897 well-defined 886:897 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 6 57 theme N-linked 860:867 arg1 sugar 876:880 only the N-linked GlcNAc sugar 851:880 only the N-linked GlcNAc sugar 851:880 However, the glycan moiety was disordered: only the N-linked GlcNAc sugar was well-defined in the electron density map. 24692304 8 58 theme protein 1144:1150 arg1 moieties 1152:1159 the protein moieties 1140:1159 the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 1140:1201 Superimposition of the structures of the protein moieties of L-Ser-CCL1 and glycosylated-L-Ser-CCL1 revealed there was no significant alteration of the protein structure by N-glycosylation. 24692304 2 59 theme oligosaccharide 308:322 arg1 moiety 333:338 the oligosaccharide (glycan) moiety 304:338 the oligosaccharide (glycan) moiety 304:338 Glycoproteins can be hard to crystallize because of the heterogeneity of the oligosaccharide (glycan) moiety. 24692304 1 60 theme chemokine 211:219 arg1 Ser-CCL1 221:228 the glycosylated chemokine Ser-CCL1 194:228 the glycosylated chemokine Ser-CCL1 194:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 0 61 theme non-glycosylated 53:68 arg1 forms 70:74 glycosylated and non-glycosylated forms 36:74 glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis 36:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 4 62 theme glycosylated 723:734 arg1 L-protein 736:744 the glycosylated L-protein 719:744 the glycosylated L-protein alone 719:750 Facile crystal formation occurred from a quasi-racemic mixture consisting of glycosylated L-protein and non-glycosylated-D-protein, while no crystals were obtained from the glycosylated L-protein alone. 24692304 7 63 theme racemic 930:936 arg1 mixture 938:944 A racemic mixture 928:944 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 928:997 A racemic mixture of the protein enantiomers L-Ser-CCL1 and D-Ser-CCL1 was also crystallized, and the structure of the true racemate was solved at a resolution of 2.7-2.15 Å. 24692304 1 64 theme Ser-CCL1 221:228 arg1 structure 181:189 the X-ray crystal structure 163:189 the X-ray crystal structure of the glycosylated chemokine Ser-CCL1 163:228 Our goal was to obtain the X-ray crystal structure of the glycosylated chemokine Ser-CCL1. 24692304 5 65 theme 2.6-2.1 Å 797:805 arg1 resolution 783:792 a resolution 781:792 a resolution of 2.6-2.1 Å 781:805 The structure was solved at a resolution of 2.6-2.1 Å. 24692304 3 66 theme total 397:401 arg1 synthesis 412:420 total chemical synthesis 397:420 total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure 397:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 24692304 0 67 gly glycosylated 36:47 arg1 forms 70:74 glycosylated and non-glycosylated forms 36:74 glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis 36:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 0 67 gly glycosylated 36:47 arg1 Ser-CCL1 93:100 the chemokine Ser-CCL1 79:100 the chemokine Ser-CCL1 prepared by total chemical synthesis 79:137 (Quasi-)racemic X-ray structures of glycosylated and non-glycosylated forms of the chemokine Ser-CCL1 prepared by total chemical synthesis. 24692304 3 68 theme defined 522:528 arg1 structure 539:547 defined covalent structure 522:547 defined covalent structure 522:547 We used glycosylated Ser-CCL1 that had been prepared by total chemical synthesis as a homogeneous compound containing an N-linked asialo biantennary nonasaccharide glycan moiety of defined covalent structure. 27956708 0 0 theme Rheumatoid 97:106 arg1 Arthritis 108:116 Rheumatoid Arthritis 97:116 Rheumatoid Arthritis 97:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 7 1 theme control 1179:1185 arg1 IgG 1187:1189 control IgG 1179:1189 control IgG isolated from the same patients 1179:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 1 2 link N-linked 224:231 arg1 glycans 233:239 N-linked glycans 224:239 N-linked glycans 224:239 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 4 3 theme F 723:723 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 1 4 theme Anti-Citrullinated 262:279 arg1 ACPA 301:304 ACPA 301:304 ACPA 301:304 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 1 4 theme Anti-Citrullinated 262:279 arg1 Antibodies 289:298 Anti-Citrullinated Protein Antibodies 262:298 Anti-Citrullinated Protein Antibodies (ACPA) 262:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 2 5 theme biological 499:508 arg1 effects 510:516 their mediated biological effects 484:516 their mediated biological effects 484:516 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 0 6 from Analysis 11:18 arg1 Arthritis 108:116 Rheumatoid Arthritis 97:116 Rheumatoid Arthritis 97:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 4 7 gly glycosylation 692:704 arg1 chains 769:774 heavy and light chains 753:774 total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 709:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 7 gly glycosylation 692:704 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 2 8 theme functions 364:372 arg1 variety 342:348 a variety 340:348 a variety of biological functions 340:372 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 2 8 theme functions 364:372 arg1 functions 364:372 biological functions 353:372 biological functions 353:372 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 7 9 attach present 1111:1117 arg2 Fab-glycans 1078:1088 Fab-glycans 1078:1088 Fab-glycans 1078:1088 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 9 attach present 1111:1117 arg1 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 9 attach present 1111:1117 arg1 % 1129:1129 over 90% 1122:1129 over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients 1122:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 1 10 theme Protein 281:287 arg1 ACPA 301:304 ACPA 301:304 ACPA 301:304 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 1 10 theme Protein 281:287 arg1 Antibodies 289:298 Anti-Citrullinated Protein Antibodies 262:298 Anti-Citrullinated Protein Antibodies (ACPA) 262:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 4 11 theme light 763:767 arg1 chains 769:774 heavy and light chains 753:774 total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 709:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 8 12 theme peripheral 1307:1316 arg1 blood 1318:1322 peripheral blood 1307:1322 peripheral blood 1307:1322 This feature was more prominent on ACPA isolated from synovial fluid compared with peripheral blood. 27956708 1 13 theme Antibodies 289:298 arg1 Fab-domain 248:257 the Fab-domain 244:257 the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA) 244:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 1 13 theme Antibodies 289:298 arg1 Antibodies 289:298 Anti-Citrullinated Protein Antibodies 262:298 Anti-Citrullinated Protein Antibodies (ACPA) 262:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 4 14 theme fragments 731:739 arg1 glycosylation 692:704 The N-linked glycosylation 679:704 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 679:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 9 15 gly hyper-sialylated 1499:1514 arg1 Fab-glycans 1516:1526 hyper-sialylated Fab-glycans 1499:1526 hyper-sialylated Fab-glycans 1499:1526 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 7 16 theme same 1209:1212 arg1 patients 1214:1221 the same patients 1205:1221 the same patients 1205:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 4 17 theme chains 769:774 arg1 glycosylation 692:704 The N-linked glycosylation 679:704 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 679:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 6 18 theme structural 957:966 arg1 analyses 968:975 The structural analyses 953:975 The structural analyses 953:975 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 3 19 theme control 556:562 arg1 IgG 565:567 noncitrulline specific (control) IgG 532:567 noncitrulline specific (control) IgG 532:567 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 9 20 theme novel 1418:1422 arg1 activities 1438:1447 novel immunological activities 1418:1447 novel immunological activities 1418:1447 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 4 21 theme mass 841:844 arg1 spectrometry 846:857 MALDI-TOF mass spectrometry 831:857 MALDI-TOF mass spectrometry 831:857 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 5 22 gly glycopeptide 921:932 arg2 glycopeptide 921:932 the glycopeptide level 917:938 the glycopeptide level 917:938 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 0 23 from Glycosylation 39:51 arg1 Arthritis 108:116 Rheumatoid Arthritis 97:116 Rheumatoid Arthritis 97:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 2 24 theme mediated 490:497 arg1 effects 510:516 their mediated biological effects 484:516 their mediated biological effects 484:516 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 2 25 theme ACPA-IgG 454:461 arg1 Fab-glycans 439:449 the Fab-glycans 435:449 the Fab-glycans of ACPA-IgG 435:461 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 4 26 theme N-linked 683:690 arg1 glycosylation 692:704 The N-linked glycosylation 679:704 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 679:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 6 27 theme sialylated 1025:1034 arg1 glycans 1036:1042 highly sialylated glycans 1018:1042 highly sialylated glycans 1018:1042 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 3 28 theme synovial 588:595 arg1 fluid 597:601 synovial fluid 588:601 synovial fluid 588:601 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 4 29 theme total 709:713 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 0 30 theme Structural 0:9 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 0:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 7 31 theme ACPA-IgG 1134:1141 arg1 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 31 theme ACPA-IgG 1134:1141 arg1 % 1129:1129 over 90% 1122:1129 over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients 1122:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 1 32 theme high 206:209 arg1 abundance 211:219 the unexpectedly high abundance 189:219 the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA) 189:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 0 33 theme Variable 23:30 arg1 Glycosylation 39:51 Variable Domain Glycosylation 23:51 Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 23:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 4 34 theme ACPA-IgG 779:786 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 34 theme ACPA-IgG 779:786 arg1 chains 769:774 heavy and light chains 753:774 total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 709:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 35 theme IgG 800:802 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 35 theme IgG 800:802 arg1 chains 769:774 heavy and light chains 753:774 total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 709:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 36 theme MALDI-TOF 831:839 arg1 spectrometry 846:857 MALDI-TOF mass spectrometry 831:857 MALDI-TOF mass spectrometry 831:857 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 4 37 theme control 792:798 arg1 IgG 800:802 control IgG 792:802 control IgG 792:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 2 38 theme structural 409:418 arg1 composition 420:430 the structural composition 405:430 the structural composition of the Fab-glycans of ACPA-IgG 405:461 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 2 39 theme Fab-glycans 439:449 arg1 composition 420:430 the structural composition 405:430 the structural composition of the Fab-glycans of ACPA-IgG 405:461 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 6 40 contain contain 1010:1016 arg1 molecules 1000:1008 ACPA-IgG molecules 991:1008 ACPA-IgG molecules 991:1008 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 6 40 contain contain 1010:1016 arg2 molecules 1000:1008 ACPA-IgG molecules 991:1008 ACPA-IgG molecules 991:1008 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 6 40 contain contain 1010:1016 arg1 Fab-domain 1053:1062 their Fab-domain 1047:1062 their Fab-domain 1047:1062 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 6 40 contain contain 1010:1016 arg2 glycans 1036:1042 highly sialylated glycans 1018:1042 highly sialylated glycans 1018:1042 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 2 41 theme N-linked 311:318 arg1 glycans 320:326 N-linked glycans 311:326 N-linked glycans 311:326 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 9 42 theme ACPA-IgG 1398:1405 arg1 ability 1387:1393 the ability 1383:1393 the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans 1383:1526 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 4 43 theme Fc 716:717 arg1 fragments 731:739 Fc and F(ab')2 fragments 716:739 Fc and F(ab')2 fragments 716:739 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 0 44 theme Glycosylation 39:51 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 0:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 9 45 theme binding 1470:1476 arg1 lectins 1487:1493 binding specific lectins 1470:1493 binding specific lectins 1470:1493 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 0 46 theme Glycans 160:166 arg1 Presence 130:137 the Presence 126:137 the Presence of Highly Sialylated Glycans 126:166 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 7 47 located present 1111:1117 arg2 Fab-glycans 1078:1088 Fab-glycans 1078:1088 Fab-glycans 1078:1088 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 47 located present 1111:1117 arg1 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 ACPA-IgG 1134:1141 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 47 located present 1111:1117 arg1 % 1129:1129 over 90% 1122:1129 over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients 1122:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 3 48 theme ACPA 611:614 arg1 patients 646:653 nine ACPA positive rheumatoid arthritis patients 606:653 nine ACPA positive rheumatoid arthritis patients 606:653 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 9 49 theme immunological 1424:1436 arg1 activities 1438:1447 novel immunological activities 1418:1447 novel immunological activities 1418:1447 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 1 50 theme N-linked 224:231 arg1 glycans 233:239 N-linked glycans 224:239 N-linked glycans 224:239 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 0 51 theme Domain 32:37 arg1 Glycosylation 39:51 Variable Domain Glycosylation 23:51 Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 23:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 0 52 theme Sialylated 149:158 arg1 Glycans 160:166 Highly Sialylated Glycans 142:166 Highly Sialylated Glycans 142:166 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 3 53 theme positive 616:623 arg1 arthritis 636:644 positive rheumatoid arthritis 616:644 nine ACPA positive rheumatoid arthritis patients 606:653 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 1 54 theme glycans 233:239 arg1 abundance 211:219 the unexpectedly high abundance 189:219 the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA) 189:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 0 55 theme Anti-Citrullinated 56:73 arg1 Antibodies 83:92 Anti-Citrullinated Protein Antibodies 56:92 Anti-Citrullinated Protein Antibodies 56:92 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 2 56 link N-linked 311:318 arg1 glycans 320:326 N-linked glycans 311:326 N-linked glycans 311:326 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 5 57 gly Fc-glycosylation 864:879 arg1 ACPA-IgG 884:891 ACPA-IgG 884:891 ACPA-IgG 884:891 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 5 57 gly Fc-glycosylation 864:879 arg1 level 934:938 the glycopeptide level 917:938 the glycopeptide level 917:938 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 5 57 gly Fc-glycosylation 864:879 arg1 IgG 897:899 IgG 897:899 IgG 897:899 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 2 58 theme biological 353:362 arg1 functions 364:372 biological functions 353:372 biological functions 353:372 As N-linked glycans can mediate a variety of biological functions, we now aimed at investigating the structural composition of the Fab-glycans of ACPA-IgG to better understand their mediated biological effects. 27956708 8 59 attach isolated 1264:1271 arg1 fluid 1287:1291 synovial fluid 1278:1291 synovial fluid 1278:1291 This feature was more prominent on ACPA isolated from synovial fluid compared with peripheral blood. 27956708 8 59 attach isolated 1264:1271 arg2 ACPA 1259:1262 ACPA 1259:1262 ACPA isolated from synovial fluid compared with peripheral blood 1259:1322 This feature was more prominent on ACPA isolated from synovial fluid compared with peripheral blood. 27956708 9 60 theme specific 1478:1485 arg1 lectins 1487:1493 binding specific lectins 1470:1493 binding specific lectins 1470:1493 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 4 61 link N-linked 683:690 arg1 glycosylation 692:704 The N-linked glycosylation 679:704 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 679:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 1 62 from abundance 211:219 arg1 Fab-domain 248:257 the Fab-domain 244:257 the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA) 244:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 1 62 from abundance 211:219 arg1 Antibodies 289:298 Anti-Citrullinated Protein Antibodies 262:298 Anti-Citrullinated Protein Antibodies (ACPA) 262:305 Recently, we showed the unexpectedly high abundance of N-linked glycans on the Fab-domain of Anti-Citrullinated Protein Antibodies (ACPA). 27956708 5 63 theme IgG 897:899 arg1 Fc-glycosylation 864:879 The Fc-glycosylation 860:879 The Fc-glycosylation of ACPA-IgG and IgG 860:899 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 3 64 from plasma 574:579 arg1 IgG 565:567 noncitrulline specific (control) IgG 532:567 noncitrulline specific (control) IgG 532:567 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 3 64 from plasma 574:579 arg1 ACPA-IgG 519:526 ACPA-IgG 519:526 ACPA-IgG 519:526 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 8 65 theme synovial 1278:1285 arg1 fluid 1287:1291 synovial fluid 1278:1291 synovial fluid 1278:1291 This feature was more prominent on ACPA isolated from synovial fluid compared with peripheral blood. 27956708 3 66 theme noncitrulline 532:544 arg1 IgG 565:567 noncitrulline specific (control) IgG 532:567 noncitrulline specific (control) IgG 532:567 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 3 67 theme patients 646:653 arg1 plasma 574:579 plasma 574:579 plasma 574:579 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 5 68 theme ACPA-IgG 884:891 arg1 Fc-glycosylation 864:879 The Fc-glycosylation 860:879 The Fc-glycosylation of ACPA-IgG and IgG 860:899 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 5 69 theme glycopeptide 921:932 arg1 level 934:938 the glycopeptide level 917:938 the glycopeptide level 917:938 The Fc-glycosylation of ACPA-IgG and IgG was analyzed at the glycopeptide level using LC-MS. 27956708 9 70 theme first 1356:1360 arg1 evidence 1362:1369 the first evidence 1352:1369 the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans 1352:1526 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 4 71 theme heavy 753:757 arg1 chains 769:774 heavy and light chains 753:774 total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG 709:802 The N-linked glycosylation of total, Fc and F(ab')2 fragments, as well as heavy and light chains of ACPA-IgG and control IgG were analyzed by UHPLC and MALDI-TOF mass spectrometry. 27956708 0 72 gly Glycosylation 39:51 arg1 Antibodies 83:92 Anti-Citrullinated Protein Antibodies 56:92 Anti-Citrullinated Protein Antibodies 56:92 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 3 73 dep purified 669:676 arg1 affinity 660:667 affinity 660:667 affinity 660:667 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 0 74 theme Antibodies 83:92 arg1 Glycosylation 39:51 Variable Domain Glycosylation 23:51 Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 23:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 0 75 from Arthritis 108:116 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis 0:116 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 3 76 theme rheumatoid 625:634 arg1 arthritis 636:644 positive rheumatoid arthritis 616:644 nine ACPA positive rheumatoid arthritis patients 606:653 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 7 77 attach isolated 1191:1198 arg1 patients 1214:1221 the same patients 1205:1221 the same patients 1205:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 7 77 attach isolated 1191:1198 arg2 IgG 1187:1189 control IgG 1179:1189 control IgG isolated from the same patients 1179:1221 Importantly, Fab-glycans were estimated to be present on over 90% of ACPA-IgG, which is five times higher than in control IgG isolated from the same patients. 27956708 3 78 theme specific 546:553 arg1 IgG 565:567 noncitrulline specific (control) IgG 532:567 noncitrulline specific (control) IgG 532:567 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 3 79 from fluid 597:601 arg1 IgG 565:567 noncitrulline specific (control) IgG 532:567 noncitrulline specific (control) IgG 532:567 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 3 79 from fluid 597:601 arg1 ACPA-IgG 519:526 ACPA-IgG 519:526 ACPA-IgG 519:526 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 0 80 theme Protein 75:81 arg1 Antibodies 83:92 Anti-Citrullinated Protein Antibodies 56:92 Anti-Citrullinated Protein Antibodies 56:92 Structural Analysis of Variable Domain Glycosylation of Anti-Citrullinated Protein Antibodies in Rheumatoid Arthritis Reveals the Presence of Highly Sialylated Glycans. 27956708 3 81 theme arthritis 636:644 arg1 patients 646:653 nine ACPA positive rheumatoid arthritis patients 606:653 nine ACPA positive rheumatoid arthritis patients 606:653 ACPA-IgG and noncitrulline specific (control) IgG from plasma and/or synovial fluid of nine ACPA positive rheumatoid arthritis patients were affinity purified. 27956708 6 82 gly sialylated 1025:1034 arg1 glycans 1036:1042 highly sialylated glycans 1018:1042 highly sialylated glycans 1018:1042 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 27956708 9 83 theme hyper-sialylated 1499:1514 arg1 Fab-glycans 1516:1526 hyper-sialylated Fab-glycans 1499:1526 hyper-sialylated Fab-glycans 1499:1526 These observations provide the first evidence pointing to the ability of ACPA-IgG to mediate novel immunological activities, for example through binding specific lectins via hyper-sialylated Fab-glycans. 27956708 6 84 theme ACPA-IgG 991:998 arg1 molecules 1000:1008 ACPA-IgG molecules 991:1008 ACPA-IgG molecules 991:1008 The structural analyses revealed that ACPA-IgG molecules contain highly sialylated glycans in their Fab-domain. 26098850 4 0 theme chromatography-electrospray 754:780 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 1 1 theme life 155:158 arg1 life 155:158 life 155:158 life 155:158 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 1 1 theme life 155:158 arg1 domains 144:150 all three domains 134:150 all three domains of life 134:158 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 5 2 with saturated 1087:1095 arg1 phosphates 1006:1015 other described archaeal dolichol phosphates 972:1015 other described archaeal dolichol phosphates 972:1015 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 3 theme isoprene 1180:1187 arg1 subunit 1189:1195 only a saturated α-position isoprene subunit 1152:1195 only a saturated α-position isoprene subunit 1152:1195 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 2 4 link N-linked 461:468 arg1 glycans 470:476 archaeal N-linked glycans 452:476 archaeal N-linked glycans characterized to date 452:498 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 4 5 theme liquid 747:752 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 0 6 theme Archaea 102:108 arg1 furiosus 121:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 2 7 attach derived 504:510 arg1 carrier 542:548 a dolichol monophosphate carrier 517:548 a dolichol monophosphate carrier 517:548 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 2 7 attach derived 504:510 arg2 glycans 470:476 archaeal N-linked glycans 452:476 archaeal N-linked glycans characterized to date 452:498 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 4 8 theme charged 856:862 arg1 phosphate 846:854 dolichol phosphate 837:854 dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species 837:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 0 9 theme Hyperthermophilic 84:100 arg1 furiosus 121:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 6 10 theme subunits 1253:1260 arg1 subunits 1253:1260 the 12-14 isoprene subunits 1234:1260 the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate 1234:1302 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 10 theme subunits 1253:1260 arg1 1-4 1227:1229 1-4 1227:1229 1-4 1227:1229 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 10 theme subunits 1253:1260 arg1 saturated 1308:1316 saturated 1308:1316 saturated 1308:1316 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 10 theme subunits 1253:1260 arg1 only 1341:1344 only 1341:1344 only 1341:1344 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 4 11 theme phosphate 846:854 arg1 existence 824:832 the existence 820:832 the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species 820:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 2 12 theme phosphorylated 294:307 arg1 dolichol 309:316 phosphorylated dolichol 294:316 phosphorylated dolichol for this role 294:330 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 6 13 theme P. 1273:1274 arg1 phosphate 1294:1302 P. furiosus dolichol phosphate 1273:1302 P. furiosus dolichol phosphate 1273:1302 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 5 14 dep P. 1065:1066 arg1 furiosus 1068:1075 furiosus 1068:1075 furiosus 1068:1075 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 4 15 theme mass 793:796 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 6 16 theme dolichol 1367:1374 arg1 phosphate 1376:1384 the longest archaeal dolichol phosphate 1346:1384 the longest archaeal dolichol phosphate described to date 1346:1402 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 17 dep P. 1273:1274 arg1 furiosus 1276:1283 furiosus 1276:1283 furiosus 1276:1283 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 2 18 theme dolichol 414:421 arg1 pyrophosphate 423:435 dolichol pyrophosphate 414:435 dolichol pyrophosphate in eukaryotes 414:449 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 0 19 theme Pyrococcus 110:119 arg1 furiosus 121:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 4 20 theme N-glycosylation 931:945 arg1 studies 920:926 in vitro studies 911:926 in vitro studies of N-glycosylation on this species 911:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 6 21 theme isoprene 1244:1251 arg1 subunits 1253:1260 the 12-14 isoprene subunits 1234:1260 the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate 1234:1302 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 5 22 theme dolichol 997:1004 arg1 phosphates 1006:1015 other described archaeal dolichol phosphates 972:1015 other described archaeal dolichol phosphates 972:1015 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 23 dep phosphodolichols 1122:1137 arg1 contrast 1101:1108 contrast 1101:1108 contrast 1101:1108 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 6 24 theme longest 1350:1356 arg1 phosphate 1376:1384 the longest archaeal dolichol phosphate 1346:1384 the longest archaeal dolichol phosphate described to date 1346:1402 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 5 25 theme ω-terminal 1029:1038 arg1 subunits 1049:1056 the α- and ω-terminal isoprene subunits 1018:1056 the α- and ω-terminal isoprene subunits of the P. furiosus lipid 1018:1081 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 25 theme ω-terminal 1029:1038 arg1 saturated 1087:1095 saturated 1087:1095 saturated 1087:1095 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 3 26 from furiosus 666:673 arg1 phosphate 619:627 glycan-charged dolichol phosphate 595:627 glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus 595:673 In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. 26098850 0 27 theme N-Linked 0:7 arg1 Glycans 9:15 N-Linked Glycans 0:15 N-Linked Glycans 0:15 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 5 28 theme isoprene 1040:1047 arg1 subunits 1049:1056 the α- and ω-terminal isoprene subunits 1018:1056 the α- and ω-terminal isoprene subunits of the P. furiosus lipid 1018:1081 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 28 theme isoprene 1040:1047 arg1 saturated 1087:1095 saturated 1087:1095 saturated 1087:1095 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 2 29 theme target 393:398 arg1 proteins 400:407 target proteins 393:407 target proteins 393:407 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 2 30 theme N-linked 461:468 arg1 glycans 470:476 archaeal N-linked glycans 452:476 archaeal N-linked glycans characterized to date 452:498 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 4 31 theme dolichol 837:844 arg1 phosphate 846:854 dolichol phosphate 837:854 dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species 837:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 2 32 theme archaeal 452:459 arg1 glycans 470:476 archaeal N-linked glycans 452:476 archaeal N-linked glycans characterized to date 452:498 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 4 33 with charged 856:862 arg1 heptasaccharide 873:887 heptasaccharide 873:887 heptasaccharide 873:887 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 5 34 theme saturated 1159:1167 arg1 subunit 1189:1195 only a saturated α-position isoprene subunit 1152:1195 only a saturated α-position isoprene subunit 1152:1195 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 35 theme eukaryal 1113:1120 arg1 phosphodolichols 1122:1137 eukaryal phosphodolichols 1113:1137 eukaryal phosphodolichols that present only a saturated α-position isoprene subunit 1113:1195 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 36 theme described 978:986 arg1 phosphates 1006:1015 other described archaeal dolichol phosphates 972:1015 other described archaeal dolichol phosphates 972:1015 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 2 37 theme single 564:569 arg1 example 571:577 a single example 562:577 a single example 562:577 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 5 38 theme α-position 1169:1178 arg1 subunit 1189:1195 only a saturated α-position isoprene subunit 1152:1195 only a saturated α-position isoprene subunit 1152:1195 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 4 39 theme Normal 722:727 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 1 40 theme glycans 205:211 arg1 assembly 193:200 the assembly 189:200 the assembly of glycans on phosphorylated polyisoprenoid carriers 189:253 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 6 41 theme archaeal 1358:1365 arg1 phosphate 1376:1384 the longest archaeal dolichol phosphate 1346:1384 the longest archaeal dolichol phosphate described to date 1346:1402 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 0 42 from Carriers 68:75 arg1 furiosus 121:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 the Hyperthermophilic Archaea Pyrococcus furiosus 80:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 4 43 theme in 911:912 arg1 studies 920:926 in vitro studies 911:926 in vitro studies of N-glycosylation on this species 911:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 4 44 dep in 911:912 arg1 vitro 914:918 vitro 914:918 vitro 914:918 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 0 45 theme Dolichol 49:56 arg1 Carriers 68:75 Highly Reduced Dolichol Phosphate Carriers 34:75 Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus 34:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 0 46 theme Phosphate 58:66 arg1 Carriers 68:75 Highly Reduced Dolichol Phosphate Carriers 34:75 Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus 34:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 3 47 theme glycan-charged 595:608 arg1 phosphate 619:627 glycan-charged dolichol phosphate 595:627 glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus 595:673 In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. 26098850 4 48 from studies 920:926 arg1 species 955:961 this species 950:961 this species 950:961 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 2 49 theme assembled 349:357 arg1 oligosaccharide 359:373 the assembled oligosaccharide 345:373 the assembled oligosaccharide 345:373 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 0 50 theme Reduced 41:47 arg1 Carriers 68:75 Highly Reduced Dolichol Phosphate Carriers 34:75 Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus 34:128 N-Linked Glycans Are Assembled on Highly Reduced Dolichol Phosphate Carriers in the Hyperthermophilic Archaea Pyrococcus furiosus. 26098850 6 51 theme dolichol 1285:1292 arg1 phosphate 1294:1302 P. furiosus dolichol phosphate 1273:1302 P. furiosus dolichol phosphate 1273:1302 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 3 52 theme dolichol 610:617 arg1 phosphate 619:627 glycan-charged dolichol phosphate 595:627 glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus 595:673 In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. 26098850 1 53 from assembly 193:200 arg1 carriers 246:253 phosphorylated polyisoprenoid carriers 216:253 phosphorylated polyisoprenoid carriers 216:253 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 4 54 theme ionization 782:791 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 6 55 dep saturated 1308:1316 arg1 subunits 1253:1260 the 12-14 isoprene subunits 1234:1260 the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate 1234:1302 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 55 dep saturated 1308:1316 arg1 1-4 1227:1229 1-4 1227:1229 1-4 1227:1229 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 55 dep saturated 1308:1316 arg1 saturated 1308:1316 saturated 1308:1316 saturated 1308:1316 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 6 55 dep saturated 1308:1316 arg1 only 1341:1344 only 1341:1344 only 1341:1344 Interestingly, an additional 1-4 of the 12-14 isoprene subunits comprising P. furiosus dolichol phosphate are saturated, making this lipid not only the longest archaeal dolichol phosphate described to date but also the most highly saturated. 26098850 3 56 theme Pyrococcus 655:664 arg1 furiosus 666:673 the hyperthermophile Pyrococcus furiosus 634:673 the hyperthermophile Pyrococcus furiosus 634:673 In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. 26098850 5 57 theme α- 1022:1023 arg1 subunits 1049:1056 the α- and ω-terminal isoprene subunits 1018:1056 the α- and ω-terminal isoprene subunits of the P. furiosus lipid 1018:1081 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 57 theme α- 1022:1023 arg1 saturated 1087:1095 saturated 1087:1095 saturated 1087:1095 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 58 theme other 972:976 arg1 phosphates 1006:1015 other described archaeal dolichol phosphates 972:1015 other described archaeal dolichol phosphates 972:1015 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 1 59 theme phosphorylated 216:229 arg1 carriers 246:253 phosphorylated polyisoprenoid carriers 216:253 phosphorylated polyisoprenoid carriers 216:253 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 5 60 theme lipid 1077:1081 arg1 subunits 1049:1056 the α- and ω-terminal isoprene subunits 1018:1056 the α- and ω-terminal isoprene subunits of the P. furiosus lipid 1018:1081 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 60 theme lipid 1077:1081 arg1 saturated 1087:1095 saturated 1087:1095 saturated 1087:1095 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 5 61 theme archaeal 988:995 arg1 phosphates 1006:1015 other described archaeal dolichol phosphates 972:1015 other described archaeal dolichol phosphates 972:1015 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 1 62 theme polyisoprenoid 231:244 arg1 carriers 246:253 phosphorylated polyisoprenoid carriers 216:253 phosphorylated polyisoprenoid carriers 216:253 In all three domains of life, N-glycosylation begins with the assembly of glycans on phosphorylated polyisoprenoid carriers. 26098850 2 63 theme monophosphate 528:540 arg1 carrier 542:548 a dolichol monophosphate carrier 517:548 a dolichol monophosphate carrier 517:548 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 2 64 from pyrophosphate 423:435 arg1 eukaryotes 440:449 eukaryotes 440:449 eukaryotes 440:449 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 4 65 theme phase 741:745 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 26098850 2 66 theme dolichol 519:526 arg1 carrier 542:548 a dolichol monophosphate carrier 517:548 a dolichol monophosphate carrier 517:548 Like eukaryotes, archaea also utilize phosphorylated dolichol for this role, yet whereas the assembled oligosaccharide is transferred to target proteins from dolichol pyrophosphate in eukaryotes, archaeal N-linked glycans characterized to date are derived from a dolichol monophosphate carrier, apart from a single example. 26098850 5 67 theme P. 1065:1066 arg1 lipid 1077:1081 the P. furiosus lipid 1061:1081 the P. furiosus lipid 1061:1081 As with other described archaeal dolichol phosphates, the α- and ω-terminal isoprene subunits of the P. furiosus lipid are saturated, in contrast to eukaryal phosphodolichols that present only a saturated α-position isoprene subunit. 26098850 3 68 theme hyperthermophile 638:653 arg1 furiosus 666:673 the hyperthermophile Pyrococcus furiosus 634:673 the hyperthermophile Pyrococcus furiosus 634:673 In this study, glycan-charged dolichol phosphate from the hyperthermophile Pyrococcus furiosus was identified and structurally characterized. 26098850 4 69 theme reverse 733:739 arg1 spectrometry 798:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry 722:809 Normal and reverse phase liquid chromatography-electrospray ionization mass spectrometry revealed the existence of dolichol phosphate charged with the heptasaccharide recently described in in vitro studies of N-glycosylation on this species. 24680513 6 0 theme structures 1253:1262 arg1 co-elution 1234:1243 the co-elution 1230:1243 the co-elution of many structures 1230:1262 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 7 1 theme techniques 1436:1445 arg1 complementarity 1400:1414 the complementarity 1396:1414 the complementarity of these separation techniques 1396:1445 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 5 2 theme resolution 873:882 arg1 methods 884:890 these high resolution methods 862:890 these high resolution methods 862:890 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 5 3 theme different 900:908 arg1 profiles 910:917 different profiles 900:917 different profiles due to diverse chemistries 900:944 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 6 4 theme many 1248:1251 arg1 structures 1253:1262 many structures 1248:1262 many structures 1248:1262 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 1 5 theme information 165:175 arg1 content 177:183 information content 165:183 information content 165:183 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 4 6 theme ovine 572:576 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 0 7 theme mammalian 93:101 arg1 species 103:109 healthy mammalian species 85:109 healthy mammalian species 85:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 4 8 theme reversed 722:729 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 8 theme reversed 722:729 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 5 9 theme high 868:871 arg1 methods 884:890 these high resolution methods 862:890 these high resolution methods 862:890 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 0 10 theme healthy 85:91 arg1 species 103:109 healthy mammalian species 85:109 healthy mammalian species 85:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 2 11 theme significant 292:302 arg1 role 304:307 a significant role 290:307 a significant role 290:307 Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. 24680513 4 12 theme equine 579:584 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 0 13 from elucidation 44:54 arg1 species 103:109 healthy mammalian species 85:109 healthy mammalian species 85:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 6 14 dep glycans 1138:1144 arg1 mannosylated 1154:1165 mannosylated 1154:1165 mannosylated 1154:1165 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 4 15 theme capillary 751:759 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 15 theme capillary 751:759 arg1 electrophoresis 761:775 capillary electrophoresis 751:775 capillary electrophoresis 751:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 5 16 theme glycans 851:857 arg1 separation 833:842 The separation 829:842 The separation of the glycans by these high resolution methods 829:890 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 4 17 link N-linked 531:538 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 6 18 theme glycans 1138:1144 arg1 classes 1127:1133 classes 1127:1133 classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures 1127:1262 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 4 19 theme separation 639:648 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 19 theme separation 639:648 arg1 electrophoresis 761:775 capillary electrophoresis 751:775 capillary electrophoresis 751:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 19 theme separation 639:648 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 19 theme separation 639:648 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 20 dep human 557:561 arg1 bovine 564:569 bovine 564:569 bovine 564:569 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 21 theme phase 731:735 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 21 theme phase 731:735 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 7 22 from complexity 1320:1329 arg1 composition 1363:1373 their N-glycan composition 1348:1373 their N-glycan composition 1348:1373 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 4 23 theme hydrophilic 662:672 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 23 theme hydrophilic 662:672 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 6 24 theme % 960:960 arg1 abundance 962:970 the % abundance 956:970 the % abundance of structures obtained by CE-LIF and HILIC-UPLC 956:1018 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 6 24 theme % 960:960 arg1 similar 1025:1031 similar 1025:1031 similar 1025:1031 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 2 25 theme biological 317:326 arg1 processes 328:336 many biological processes 312:336 many biological processes 312:336 Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. 24680513 0 26 theme techniques 25:34 arg1 Comparison 0:9 Comparison 0:9 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 0:110 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 4 27 theme human 557:561 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 2 28 theme many 312:315 arg1 processes 328:336 many biological processes 312:336 many biological processes 312:336 Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. 24680513 7 29 from variation 1335:1343 arg1 composition 1363:1373 their N-glycan composition 1348:1373 their N-glycan composition 1348:1373 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 0 30 theme separation 14:23 arg1 techniques 25:34 separation techniques 14:34 separation techniques 14:34 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 5 31 theme diverse 926:932 arg1 chemistries 934:944 diverse chemistries 926:944 diverse chemistries 926:944 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 4 32 theme chromatography 693:706 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 32 theme chromatography 693:706 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 1 33 theme excellent 200:208 arg1 source 210:215 an excellent source 197:215 an excellent source for biomarker discovery in mammalian health 197:259 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 4 34 theme HILIC 709:713 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 34 theme HILIC 709:713 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 6 35 dep mannosylated 1154:1165 arg1 fucosylated 1191:1201 fucosylated 1191:1201 fucosylated 1191:1201 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 6 35 dep mannosylated 1154:1165 arg1 bisected 1181:1188 bisected 1181:1188 bisected 1181:1188 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 6 35 dep mannosylated 1154:1165 arg1 fucosylated 1168:1178 fucosylated 1168:1178 fucosylated 1168:1178 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 6 35 dep mannosylated 1154:1165 arg1 bisected 1207:1214 bisected 1207:1214 bisected 1207:1214 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 7 36 theme N-glycan 1354:1361 arg1 composition 1363:1373 their N-glycan composition 1348:1373 their N-glycan composition 1348:1373 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 4 37 theme glycan 632:637 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 37 theme glycan 632:637 arg1 electrophoresis 761:775 capillary electrophoresis 751:775 capillary electrophoresis 751:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 37 theme glycan 632:637 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 37 theme glycan 632:637 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 38 theme liquid 686:691 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 38 theme liquid 686:691 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 39 theme N-linked 531:538 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 6 40 theme structures 975:984 arg1 abundance 962:970 the % abundance 956:970 the % abundance of structures obtained by CE-LIF and HILIC-UPLC 956:1018 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 6 40 theme structures 975:984 arg1 similar 1025:1031 similar 1025:1031 similar 1025:1031 However, the % abundance of structures obtained by CE-LIF and HILIC-UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. 24680513 4 41 theme fluorescence 796:807 arg1 detection 809:817 fluorescence detection 796:817 fluorescence detection (CE-LIF) 796:826 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 41 theme fluorescence 796:807 arg1 CE-LIF 820:825 CE-LIF 820:825 CE-LIF 820:825 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 42 theme orthogonal 621:630 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 42 theme orthogonal 621:630 arg1 electrophoresis 761:775 capillary electrophoresis 751:775 capillary electrophoresis 751:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 42 theme orthogonal 621:630 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 42 theme orthogonal 621:630 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 7 43 theme separation 1425:1434 arg1 techniques 1436:1445 these separation techniques 1419:1445 these separation techniques 1419:1445 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 1 44 theme IgG 116:118 arg1 N-glycome 120:128 The IgG N-glycome 112:128 The IgG N-glycome 112:128 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 4 45 theme glycosylation 540:552 arg1 variation 518:526 the variation 514:526 the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 514:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 0 46 from species 103:109 arg1 elucidation 44:54 the elucidation 40:54 the elucidation of IgG N-glycans pooled from healthy mammalian species 40:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 0 46 from species 103:109 arg1 N-glycans 63:71 IgG N-glycans 59:71 IgG N-glycans pooled from healthy mammalian species 59:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 4 47 theme RP 738:739 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 47 theme RP 738:739 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 1 48 theme biomarker 221:229 arg1 discovery 231:239 biomarker discovery 221:239 biomarker discovery in mammalian health 221:259 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 1 49 from discovery 231:239 arg1 health 254:259 mammalian health 244:259 mammalian health 244:259 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 5 50 theme due 919:921 arg1 profiles 910:917 different profiles 900:917 different profiles due to diverse chemistries 900:944 The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. 24680513 7 51 theme various 1279:1285 arg1 species 1287:1293 various species 1279:1293 various species 1279:1293 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 4 52 theme canine 587:592 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 0 53 theme N-glycans 63:71 arg1 elucidation 44:54 the elucidation 40:54 the elucidation of IgG N-glycans pooled from healthy mammalian species 40:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 4 54 dep techniques 650:659 arg1 -UPLC 741:745 reversed phase (RP)-UPLC 722:745 reversed phase (RP)-UPLC 722:745 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 54 dep techniques 650:659 arg1 electrophoresis 761:775 capillary electrophoresis 751:775 capillary electrophoresis 751:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 54 dep techniques 650:659 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 54 dep techniques 650:659 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 7 55 from species 1287:1293 arg1 IgGs 1269:1272 The IgGs 1265:1272 The IgGs from various species 1265:1293 The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques. 24680513 1 56 theme sufficient 139:148 arg1 complexity 150:159 sufficient complexity 139:159 sufficient complexity 139:159 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 24680513 4 57 theme IgG 605:607 arg1 glycosylation 540:552 N-linked glycosylation 531:552 N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG 531:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 0 58 theme IgG 59:61 arg1 N-glycans 63:71 IgG N-glycans 59:71 IgG N-glycans pooled from healthy mammalian species 59:109 Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species. 24680513 4 59 theme feline 598:603 arg1 IgG 605:607 feline IgG 598:607 feline IgG 598:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 60 gly glycosylation 540:552 arg1 equine 579:584 equine 579:584 equine 579:584 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 60 gly glycosylation 540:552 arg1 ovine 572:576 ovine 572:576 ovine 572:576 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 60 gly glycosylation 540:552 arg1 human 557:561 human 557:561 human 557:561 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 60 gly glycosylation 540:552 arg1 canine 587:592 canine 587:592 canine 587:592 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 60 gly glycosylation 540:552 arg1 IgG 605:607 feline IgG 598:607 feline IgG 598:607 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 61 theme interaction 674:684 arg1 techniques 650:659 three orthogonal glycan separation techniques 615:659 three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis 615:775 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 4 61 theme interaction 674:684 arg1 -UPLC 715:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 hydrophilic interaction liquid chromatography (HILIC)-UPLC 662:719 We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)-UPLC, reversed phase (RP)-UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). 24680513 1 62 theme mammalian 244:252 arg1 health 254:259 mammalian health 244:259 mammalian health 244:259 The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. 26648038 0 0 theme bitter 65:70 arg1 gourd 72:76 bitter gourd 65:76 bitter gourd 65:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 5 1 theme binding 651:657 arg1 site 659:662 a sugar binding site 643:662 a sugar binding site 643:662 BGSL has a sugar binding site only on one domain, thus impairing its interaction at the cell surface. 26648038 9 2 theme selection 1132:1140 arg1 case 1109:1112 a case 1107:1112 a case of conformational selection 1107:1140 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 4 3 theme type 601:604 arg1 RIPs 609:612 type II RIPs 601:612 type II RIPs 601:612 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 8 4 from loop 1032:1035 arg1 chain 1051:1055 the lectin chain 1040:1055 the lectin chain 1040:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 0 5 from studies 11:17 arg1 homologue 34:42 a non-toxic homologue 22:42 a non-toxic homologue of type II RIPs from bitter gourd 22:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 8 6 theme conformations 1005:1017 arg1 presence 967:974 the presence 963:974 the presence of two possible well defined conformations of a surface loop in the lectin chain 963:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 4 7 from site 593:596 arg1 RIPs 609:612 type II RIPs 601:612 type II RIPs 601:612 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 6 8 from site 756:759 arg1 chain 778:782 the catalytic chain 764:782 the catalytic chain 764:782 The adenine binding site in the catalytic chain is defective. 26648038 6 9 theme catalytic 768:776 arg1 chain 778:782 the catalytic chain 764:782 the catalytic chain 764:782 The adenine binding site in the catalytic chain is defective. 26648038 0 10 dep basis 89:93 arg1 studies 11:17 Structural studies 0:17 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd 0:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 8 11 theme defined 997:1003 arg1 conformations 1005:1017 two possible well defined conformations 979:1017 two possible well defined conformations of a surface loop in the lectin chain 979:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 10 12 theme known 1369:1373 arg1 structure 1375:1383 known structure 1369:1383 known structure 1369:1383 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 5 13 contain has 639:641 arg1 BGSL 634:637 BGSL 634:637 BGSL 634:637 BGSL has a sugar binding site only on one domain, thus impairing its interaction at the cell surface. 26648038 5 13 contain has 639:641 arg2 site 659:662 a sugar binding site 643:662 a sugar binding site 643:662 BGSL has a sugar binding site only on one domain, thus impairing its interaction at the cell surface. 26648038 9 14 theme arginyl 1237:1243 arg1 residue 1245:1251 an arginyl residue 1234:1251 an arginyl residue in the loop 1234:1263 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 8 15 from chain 1051:1055 arg1 conformations 1005:1017 two possible well defined conformations 979:1017 two possible well defined conformations of a surface loop in the lectin chain 979:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 7 16 theme sugar 815:819 arg1 binding 821:827 sugar binding 815:827 sugar binding as well as adenine binding 815:854 Thus, defects in sugar binding as well as adenine binding appear to contribute to the non-toxicity of the lectin. 26648038 0 17 theme Molecular 79:87 arg1 basis 89:93 Molecular basis 79:93 Molecular basis of non-toxicity, conformational selection and glycan structure 79:156 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 9 18 theme conformational 1117:1130 arg1 selection 1132:1140 conformational selection 1117:1140 conformational selection 1117:1140 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 9 19 theme sugar 1087:1091 arg1 complexes 1093:1101 the sugar complexes 1083:1101 the sugar complexes 1083:1101 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 0 20 theme Structural 0:9 arg1 studies 11:17 Structural studies 0:17 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd 0:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 10 21 theme lectin 1293:1298 arg1 N-glycosylation 1270:1284 The N-glycosylation 1266:1284 The N-glycosylation of the lectin 1266:1298 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 8 22 theme surface 1024:1030 arg1 loop 1032:1035 a surface loop 1022:1035 a surface loop in the lectin chain 1022:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 6 23 theme binding 748:754 arg1 site 756:759 The adenine binding site 736:759 The adenine binding site in the catalytic chain 736:782 The adenine binding site in the catalytic chain is defective. 26648038 6 23 theme binding 748:754 arg1 defective 787:795 defective 787:795 defective 787:795 The adenine binding site in the catalytic chain is defective. 26648038 10 24 theme II 1358:1359 arg1 RIPs 1361:1364 toxic type II RIPs 1347:1364 toxic type II RIPs of known structure 1347:1383 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 2 25 theme disulphide 487:496 arg1 bridge 498:503 a disulphide bridge 485:503 a disulphide bridge 485:503 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 4 26 theme carbohydrate 572:583 arg1 structure 623:631 known structure 617:631 known structure 617:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 26 theme carbohydrate 572:583 arg1 site 593:596 a carbohydrate binding site 570:596 a carbohydrate binding site in type II RIPs of known structure 570:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 1 27 theme sugar 286:290 arg1 complexes 292:300 its sugar complexes 282:300 its sugar complexes 282:300 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 4 28 contain carries 562:568 arg2 structure 623:631 known structure 617:631 known structure 617:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 28 contain carries 562:568 arg2 site 593:596 a carbohydrate binding site 570:596 a carbohydrate binding site in type II RIPs of known structure 570:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 28 contain carries 562:568 arg1 domain 555:560 Each domain 550:560 Each domain 550:560 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 29 theme structure 623:631 arg1 structure 623:631 known structure 617:631 known structure 617:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 29 theme structure 623:631 arg1 site 593:596 a carbohydrate binding site 570:596 a carbohydrate binding site in type II RIPs of known structure 570:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 8 30 theme possible 983:990 arg1 conformations 1005:1017 two possible well defined conformations 979:1017 two possible well defined conformations of a surface loop in the lectin chain 979:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 1 31 theme non-toxic 241:249 arg1 homologue 251:259 a non-toxic homologue 239:259 a non-toxic homologue of type II RIPs 239:275 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 31 theme non-toxic 241:249 arg1 gourd 213:217 bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 206:276 gourd 213:217 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 0 32 theme non-toxicity 98:109 arg1 basis 89:93 Molecular basis 79:93 Molecular basis of non-toxicity, conformational selection and glycan structure 79:156 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 2 33 theme symmetric 349:357 arg1 protein 360:366 protein 360:366 protein 360:366 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 2 33 theme symmetric 349:357 arg1 four-chain 328:337 The four-chain 324:337 The four-chain 324:337 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 8 34 theme loop 1032:1035 arg1 conformations 1005:1017 two possible well defined conformations 979:1017 two possible well defined conformations of a surface loop in the lectin chain 979:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 7 35 theme lectin 904:909 arg1 non-toxicity 884:895 the non-toxicity 880:895 the non-toxicity of the lectin 880:909 Thus, defects in sugar binding as well as adenine binding appear to contribute to the non-toxicity of the lectin. 26648038 6 36 theme adenine 740:746 arg1 site 756:759 The adenine binding site 736:759 The adenine binding site in the catalytic chain 736:782 The adenine binding site in the catalytic chain is defective. 26648038 6 36 theme adenine 740:746 arg1 defective 787:795 defective 787:795 defective 787:795 The adenine binding site in the catalytic chain is defective. 26648038 10 37 theme type 1353:1356 arg1 RIPs 1361:1364 toxic type II RIPs 1347:1364 toxic type II RIPs of known structure 1347:1383 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 1 38 theme independent 182:192 arg1 crystals 194:201 nine independent crystals 177:201 nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 177:276 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 8 39 theme lectin 1044:1049 arg1 chain 1051:1055 the lectin chain 1040:1055 the lectin chain 1040:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 9 40 theme chosen 1150:1155 arg1 conformation 1157:1168 the chosen conformation 1146:1168 the chosen conformation 1146:1168 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 0 41 theme conformational 112:125 arg1 selection 127:135 conformational selection 112:135 conformational selection 112:135 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 3 42 theme lectin 510:515 arg1 chain 517:521 The lectin chain 506:521 The lectin chain 506:521 The lectin chain is made up of two domains. 26648038 1 43 theme crystals 194:201 arg1 structures 163:172 The structures 159:172 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 159:276 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 43 theme crystals 194:201 arg1 complexes 292:300 its sugar complexes 282:300 its sugar complexes 282:300 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 0 44 theme non-toxic 24:32 arg1 homologue 34:42 a non-toxic homologue 22:42 a non-toxic homologue of type II RIPs from bitter gourd 22:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 1 45 theme type 264:267 arg1 RIPs 272:275 type II RIPs 264:275 type II RIPs 264:275 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 2 46 theme lectin 458:463 arg1 chain 465:469 a lectin chain 456:469 a lectin chain 456:469 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 9 47 theme them 1065:1068 arg1 them 1065:1068 them 1065:1068 them 1065:1068 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 9 47 theme them 1065:1068 arg1 One 1058:1060 One 1058:1060 One 1058:1060 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 5 48 theme cell 722:725 arg1 surface 727:733 the cell surface 718:733 the cell surface 718:733 BGSL has a sugar binding site only on one domain, thus impairing its interaction at the cell surface. 26648038 2 49 theme catalytic 436:444 arg1 chain 446:450 a catalytic chain 434:450 a catalytic chain 434:450 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 0 50 theme type 47:50 arg1 RIPs 55:58 type II RIPs 47:58 type II RIPs 47:58 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 1 51 theme II 269:270 arg1 RIPs 272:275 type II RIPs 264:275 type II RIPs 264:275 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 10 52 theme structure 1375:1383 arg1 RIPs 1361:1364 toxic type II RIPs 1347:1364 toxic type II RIPs of known structure 1347:1383 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 1 53 dep gourd 213:217 arg1 BGSL 232:235 BGSL 232:235 BGSL 232:235 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 53 dep gourd 213:217 arg1 lectin 224:229 seed lectin 219:229 seed lectin (BGSL) 219:236 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 2 54 theme identical 386:394 arg1 each 415:418 each 415:418 each 415:418 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 2 54 theme identical 386:394 arg1 modules 406:412 two identical two-chain modules 382:412 two identical two-chain modules 382:412 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 1 55 theme RIPs 272:275 arg1 homologue 251:259 a non-toxic homologue 239:259 a non-toxic homologue of type II RIPs 239:275 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 55 theme RIPs 272:275 arg1 gourd 213:217 bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 206:276 gourd 213:217 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 0 56 theme selection 127:135 arg1 basis 89:93 Molecular basis 79:93 Molecular basis of non-toxicity, conformational selection and glycan structure 79:156 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 10 57 theme toxic 1347:1351 arg1 RIPs 1361:1364 toxic type II RIPs 1347:1364 toxic type II RIPs of known structure 1347:1383 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 10 58 gly N-glycosylation 1270:1284 arg1 lectin 1293:1298 the lectin 1289:1298 the lectin 1289:1298 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26648038 9 59 theme additional 1185:1194 arg1 interaction 1196:1206 an additional interaction 1182:1206 an additional interaction with the sugar 1182:1221 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 0 60 theme structure 148:156 arg1 basis 89:93 Molecular basis 79:93 Molecular basis of non-toxicity, conformational selection and glycan structure 79:156 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 8 61 from conformations 1005:1017 arg1 chain 1051:1055 the lectin chain 1040:1055 the lectin chain 1040:1055 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 5 62 theme sugar 645:649 arg1 site 659:662 a sugar binding site 643:662 a sugar binding site 643:662 BGSL has a sugar binding site only on one domain, thus impairing its interaction at the cell surface. 26648038 4 63 theme known 617:621 arg1 structure 623:631 known structure 617:631 known structure 617:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 9 64 with interaction 1196:1206 arg1 sugar 1217:1221 the sugar 1213:1221 the sugar 1213:1221 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 1 65 theme bitter 206:211 arg1 homologue 251:259 a non-toxic homologue 239:259 a non-toxic homologue of type II RIPs 239:275 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 65 theme bitter 206:211 arg1 gourd 213:217 bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 206:276 gourd 213:217 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 0 66 theme glycan 141:146 arg1 structure 148:156 glycan structure 141:156 glycan structure 141:156 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 7 67 theme adenine 840:846 arg1 binding 848:854 adenine binding 840:854 sugar binding as well as adenine binding 815:854 Thus, defects in sugar binding as well as adenine binding appear to contribute to the non-toxicity of the lectin. 26648038 7 68 from defects 804:810 arg1 binding 848:854 adenine binding 840:854 sugar binding as well as adenine binding 815:854 Thus, defects in sugar binding as well as adenine binding appear to contribute to the non-toxicity of the lectin. 26648038 7 68 from defects 804:810 arg1 binding 821:827 sugar binding 815:827 sugar binding as well as adenine binding 815:854 Thus, defects in sugar binding as well as adenine binding appear to contribute to the non-toxicity of the lectin. 26648038 1 69 theme gourd 213:217 arg1 crystals 194:201 nine independent crystals 177:201 nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, 177:276 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 0 70 theme RIPs 55:58 arg1 homologue 34:42 a non-toxic homologue 22:42 a non-toxic homologue of type II RIPs from bitter gourd 22:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 0 71 from gourd 72:76 arg1 homologue 34:42 a non-toxic homologue 22:42 a non-toxic homologue of type II RIPs from bitter gourd 22:76 Structural studies on a non-toxic homologue of type II RIPs from bitter gourd: Molecular basis of non-toxicity, conformational selection and glycan structure. 26648038 1 72 theme seed 219:222 arg1 BGSL 232:235 BGSL 232:235 BGSL 232:235 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 1 72 theme seed 219:222 arg1 lectin 224:229 seed lectin 219:229 seed lectin (BGSL) 219:236 The structures of nine independent crystals of bitter gourd seed lectin (BGSL), a non-toxic homologue of type II RIPs, and its sugar complexes have been determined. 26648038 2 73 theme two-chain 396:404 arg1 each 415:418 each 415:418 each 415:418 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 2 73 theme two-chain 396:404 arg1 modules 406:412 two identical two-chain modules 382:412 two identical two-chain modules 382:412 The four-chain, two-fold symmetric, protein is made up of two identical two-chain modules, each consisting of a catalytic chain and a lectin chain, connected by a disulphide bridge. 26648038 9 74 from residue 1245:1251 arg1 loop 1260:1263 the loop 1256:1263 the loop 1256:1263 One of them is chosen in the sugar complexes, in a case of conformational selection, as the chosen conformation facilitates an additional interaction with the sugar, involving an arginyl residue in the loop. 26648038 4 75 theme binding 585:591 arg1 structure 623:631 known structure 617:631 known structure 617:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 4 75 theme binding 585:591 arg1 site 593:596 a carbohydrate binding site 570:596 a carbohydrate binding site in type II RIPs of known structure 570:631 Each domain carries a carbohydrate binding site in type II RIPs of known structure. 26648038 8 76 theme molecule 934:941 arg1 plasticity 916:925 The plasticity 912:925 The plasticity of the molecule 912:941 The plasticity of the molecule is mainly caused by the presence of two possible well defined conformations of a surface loop in the lectin chain. 26648038 10 77 theme plant-specific 1311:1324 arg1 glycan 1326:1331 a plant-specific glycan 1309:1331 a plant-specific glycan 1309:1331 The N-glycosylation of the lectin involves a plant-specific glycan while that in toxic type II RIPs of known structure involves a glycan which is animal as well as plant specific. 26067753 2 0 theme expression 442:451 arg1 platform 453:460 a plant-based expression platform 428:460 a plant-based expression platform 428:460 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 1 1 dep glycosylation 261:273 arg1 addition 232:239 addition 232:239 addition 232:239 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 2 theme monoclonal 150:159 arg1 CxMab 181:185 CxMab 181:185 CxMab 181:185 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 2 theme monoclonal 150:159 arg1 reporter 281:288 a reporter 279:288 a reporter 279:288 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 2 theme monoclonal 150:159 arg1 antibody 161:168 the monoclonal antibody 146:168 the monoclonal antibody cetuximab (CxMab) 146:186 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 0 3 theme core 64:67 arg1 fucosylation 69:80 core fucosylation 64:80 core fucosylation 64:80 Processing of complex N-glycans in IgG Fc-region is affected by core fucosylation. 26067753 6 4 theme strong 851:856 arg1 support 858:864 A particularly strong support 836:864 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose 836:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 7 5 theme carbohydrate 1032:1043 arg1 chain 1045:1049 the Fc carbohydrate chain 1025:1049 the Fc carbohydrate chain 1025:1049 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 3 6 theme GnGn 469:472 arg1 variants 504:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 0 7 from Processing 0:9 arg1 Fc-region 39:47 IgG Fc-region 35:47 IgG Fc-region 35:47 Processing of complex N-glycans in IgG Fc-region is affected by core fucosylation. 26067753 1 8 gly glycosite 201:209 arg2 glycosite 201:209 a glycosite 199:209 a glycosite in the Fab domain 199:227 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 5 9 theme fucose 828:833 arg1 presence 811:818 the presence 807:818 the presence of core fucose 807:833 By contrast, Fc glycan processing largely depend on the presence of core fucose. 26067753 3 10 theme GnGnF 489:493 arg1 variants 504:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 3 11 dep diversification 588:602 arg1 bisected 605:612 bisected 605:612 bisected 605:612 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 3 11 dep diversification 588:602 arg1 structures 628:637 branching structures 618:637 branching structures 618:637 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 8 12 theme therapeutic 1293:1303 arg1 antibodies 1305:1314 glycan-engineered therapeutic antibodies 1275:1314 glycan-engineered therapeutic antibodies with improved efficacies 1275:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 6 13 from support 858:864 arg1 presence 894:901 the presence 890:901 the presence of plant-specific core α1,3-fucose 890:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 8 14 theme glycan-engineered 1275:1291 arg1 antibodies 1305:1314 glycan-engineered therapeutic antibodies 1275:1314 glycan-engineered therapeutic antibodies with improved efficacies 1275:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 4 15 theme Mass 641:644 arg1 analyses 672:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 4 16 theme glycan 665:670 arg1 analyses 672:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 8 17 theme IgG 1209:1211 arg1 antibodies 1213:1222 IgG antibodies 1209:1222 IgG antibodies 1209:1222 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 4 18 theme glycans 718:724 arg1 processing 700:709 efficient processing 690:709 efficient processing of Fab glycans toward envisaged structures 690:752 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 8 19 theme glycan 1188:1193 arg1 processing 1195:1204 glycan processing 1188:1204 glycan processing 1188:1204 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 6 20 theme α1,3-fucose 926:936 arg1 presence 894:901 the presence 890:901 the presence of plant-specific core α1,3-fucose 890:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 4 21 theme Fab 714:716 arg1 glycans 718:724 Fab glycans 714:724 Fab glycans 714:724 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 2 22 theme CxMab 348:352 arg1 glycoforms 334:343 bi-antennary glycoforms 321:343 bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) 321:408 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 2 23 theme plant-based 430:440 arg1 platform 453:460 a plant-based expression platform 428:460 a plant-based expression platform 428:460 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 2 24 gly glycoforms 334:343 arg1 CxMab 348:352 CxMab 348:352 CxMab 348:352 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 6 25 theme core 921:924 arg1 α1,3-fucose 926:936 plant-specific core α1,3-fucose 906:936 plant-specific core α1,3-fucose 906:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 7 26 theme chain 1045:1049 arg1 interactions 1009:1020 the interactions 1005:1020 the interactions of the Fc carbohydrate chain 1005:1049 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 1 27 dep antibody 161:168 arg1 cetuximab 170:178 cetuximab 170:178 the monoclonal antibody cetuximab (CxMab) 146:186 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 8 28 theme improved 1321:1328 arg1 efficacies 1330:1339 improved efficacies 1321:1339 improved efficacies 1321:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 7 29 theme core 1080:1083 arg1 fucose 1085:1090 core fucose 1080:1090 core fucose 1080:1090 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 0 30 theme N-glycans 22:30 arg1 Processing 0:9 Processing 0:9 Processing of complex N-glycans in IgG Fc-region 0:47 Processing of complex N-glycans in IgG Fc-region is affected by core fucosylation. 26067753 1 31 theme conserved 248:256 arg1 glycosylation 261:273 the conserved Fc glycosylation 244:273 the conserved Fc glycosylation 244:273 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 3 32 theme CxMab 498:502 arg1 variants 504:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 4 33 theme efficient 690:698 arg1 processing 700:709 efficient processing 690:709 efficient processing of Fab glycans toward envisaged structures 690:752 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 5 34 theme glycan 771:776 arg1 processing 778:787 Fc glycan processing 768:787 Fc glycan processing 768:787 By contrast, Fc glycan processing largely depend on the presence of core fucose. 26067753 0 35 theme complex 14:20 arg1 N-glycans 22:30 complex N-glycans 14:30 complex N-glycans 14:30 Processing of complex N-glycans in IgG Fc-region is affected by core fucosylation. 26067753 1 36 theme Fc 258:259 arg1 glycosylation 261:273 the conserved Fc glycosylation 244:273 the conserved Fc glycosylation 244:273 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 7 37 theme Fc 1029:1030 arg1 chain 1045:1049 the Fc carbohydrate chain 1025:1049 the Fc carbohydrate chain 1025:1049 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 1 38 theme N-glycan 99:106 arg1 processing 108:117 N-glycan processing 99:117 N-glycan processing of immunoglobulin G1 99:138 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 5 39 theme core 823:826 arg1 fucose 828:833 core fucose 823:833 core fucose 823:833 By contrast, Fc glycan processing largely depend on the presence of core fucose. 26067753 0 40 theme IgG 35:37 arg1 Fc-region 39:47 IgG Fc-region 35:47 IgG Fc-region 35:47 Processing of complex N-glycans in IgG Fc-region is affected by core fucosylation. 26067753 8 41 theme processing 1195:1204 arg1 mechanisms 1174:1183 molecular mechanisms 1164:1183 molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies 1164:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 2 42 theme core 367:370 arg1 fucosylation 372:383 core fucosylation 367:383 core fucosylation (α1,3- and α1,6-linkage) 367:408 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 3 43 theme further 539:545 arg1 processing 547:556 further processing 539:556 further processing toward sialylation and GlcNAc diversification (bisected and branching structures) 539:638 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 7 44 theme molecular 966:974 arg1 modeling 976:983 molecular modeling 966:983 molecular modeling 966:983 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 7 45 theme fucose 1085:1090 arg1 presence 1068:1075 the presence 1064:1075 the presence of core fucose 1064:1090 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 6 46 theme plant-specific 906:919 arg1 α1,3-fucose 926:936 plant-specific core α1,3-fucose 906:936 plant-specific core α1,3-fucose 906:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 7 47 from changes 994:1000 arg1 interactions 1009:1020 the interactions 1005:1020 the interactions of the Fc carbohydrate chain 1005:1049 Consistently, molecular modeling suggests changes in the interactions of the Fc carbohydrate chain depending on the presence of core fucose, possibly changing the accessibility. 26067753 8 48 theme molecular 1164:1172 arg1 mechanisms 1174:1183 molecular mechanisms 1164:1183 molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies 1164:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 6 49 theme processing 876:885 arg1 support 858:864 A particularly strong support 836:864 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose 836:936 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 2 50 dep fucosylation 372:383 arg1 α1,6-linkage 396:407 α1,6-linkage 396:407 α1,6-linkage 396:407 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 2 50 dep fucosylation 372:383 arg1 α1,3- 386:390 α1,3- 386:390 α1,3- 386:390 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 6 51 theme glycan 869:874 arg1 processing 876:885 glycan processing 869:885 glycan processing 869:885 A particularly strong support of glycan processing in the presence of plant-specific core α1,3-fucose was observed. 26067753 1 52 from glycosite 201:209 arg1 domain 222:227 the Fab domain 214:227 the Fab domain 214:227 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 53 contain has 195:197 arg1 CxMab 181:185 CxMab 181:185 CxMab 181:185 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 53 contain has 195:197 arg2 glycosite 201:209 a glycosite 199:209 a glycosite in the Fab domain 199:227 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 53 contain has 195:197 arg1 reporter 281:288 a reporter 279:288 a reporter 279:288 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 1 53 contain has 195:197 arg1 antibody 161:168 the monoclonal antibody 146:168 the monoclonal antibody cetuximab (CxMab) 146:186 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 4 54 theme envisaged 733:741 arg1 structures 743:752 envisaged structures 733:752 envisaged structures 733:752 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 1 55 theme immunoglobulin 122:135 arg1 G1 137:138 immunoglobulin G1 122:138 immunoglobulin G1 122:138 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 8 56 contain have 1235:1238 arg1 processing 1195:1204 glycan processing 1188:1204 glycan processing 1188:1204 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 8 56 contain have 1235:1238 arg2 implications 1240:1251 implications 1240:1251 implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies 1240:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 8 57 theme antibodies 1305:1314 arg1 generation 1261:1270 the generation 1257:1270 the generation of glycan-engineered therapeutic antibodies with improved efficacies 1257:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 1 58 theme G1 137:138 arg1 processing 108:117 N-glycan processing 99:117 N-glycan processing of immunoglobulin G1 99:138 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26067753 4 59 theme spectrometry-based 646:663 arg1 analyses 672:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses 641:679 Mass spectrometry-based glycan analyses revealed efficient processing of Fab glycans toward envisaged structures. 26067753 3 60 theme GlcNAc 581:586 arg1 diversification 588:602 GlcNAc diversification 581:602 GlcNAc diversification (bisected and branching structures) 581:638 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 8 61 theme antibodies 1213:1222 arg1 processing 1195:1204 glycan processing 1188:1204 glycan processing 1188:1204 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 2 62 theme bi-antennary 321:332 arg1 glycoforms 334:343 bi-antennary glycoforms 321:343 bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) 321:408 Three GlcNAc (Gn) terminating bi-antennary glycoforms of CxMab differing in core fucosylation (α1,3- and α1,6-linkage) were generated in a plant-based expression platform. 26067753 8 63 with antibodies 1305:1314 arg1 efficacies 1330:1339 improved efficacies 1321:1339 improved efficacies 1321:1339 Here, we provide data that reveal molecular mechanisms of glycan processing of IgG antibodies, which may have implications for the generation of glycan-engineered therapeutic antibodies with improved efficacies. 26067753 3 64 theme branching 618:626 arg1 structures 628:637 branching structures 618:637 branching structures 618:637 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 5 65 theme Fc 768:769 arg1 processing 778:787 Fc glycan processing 768:787 Fc glycan processing 768:787 By contrast, Fc glycan processing largely depend on the presence of core fucose. 26067753 3 66 theme GnGnF 475:479 arg1 variants 504:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants 463:511 These GnGn, GnGnF(3), and GnGnF(6) CxMab variants were subjected in vivo to further processing toward sialylation and GlcNAc diversification (bisected and branching structures). 26067753 1 67 theme Fab 218:220 arg1 domain 222:227 the Fab domain 214:227 the Fab domain 214:227 We investigated N-glycan processing of immunoglobulin G1 using the monoclonal antibody cetuximab (CxMab), which has a glycosite in the Fab domain in addition to the conserved Fc glycosylation, as a reporter. 26616285 5 0 theme Hamster 783:789 arg1 cells 803:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 5 1 theme Ovary 791:795 arg1 cells 803:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 6 2 theme glycan 935:940 arg1 analysis 942:949 glycan analysis 935:949 glycan analysis 935:949 Our results demonstrated QUANTITY is an invaluable technique for glycan analysis and bioengineering. 26616285 2 3 theme chemical 302:309 arg1 properties 311:320 their chemical properties 296:320 their chemical properties 296:320 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 4 4 from samples 691:697 arg1 analysis 657:664 high-throughput glycomic analysis 632:664 high-throughput glycomic analysis from multiple biological samples 632:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 5 dep QUANTITY 460:467 arg1 Tag 507:509 Quaternary Amine Containing Isobaric Tag 470:509 Quaternary Amine Containing Isobaric Tag for Glycan 470:520 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 2 6 from level--however 196:209 arg1 glycomics--analysis 155:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 4 7 theme Containing 487:496 arg1 Tag 507:509 Quaternary Amine Containing Isobaric Tag 470:509 Quaternary Amine Containing Isobaric Tag for Glycan 470:520 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 8 theme glycans 587:593 arg1 detection 574:582 detection 574:582 detection of glycans by mass spectrometry 574:614 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 9 theme Amine 481:485 arg1 Tag 507:509 Quaternary Amine Containing Isobaric Tag 470:509 Quaternary Amine Containing Isobaric Tag for Glycan 470:520 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 5 10 theme cells 803:807 arg1 survey 751:756 glycomic survey 742:756 glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation 742:867 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 3 11 theme glycan 402:407 arg1 analysis 409:416 global glycan analysis 395:416 global glycan analysis 395:416 As a result, technologies that can facilitate global glycan analysis are highly sought after. 26616285 4 12 theme Isobaric 498:505 arg1 Tag 507:509 Quaternary Amine Containing Isobaric Tag 470:509 Quaternary Amine Containing Isobaric Tag for Glycan 470:520 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 6 13 theme invaluable 910:919 arg1 technique 921:929 an invaluable technique 907:929 an invaluable technique for glycan analysis and bioengineering 907:968 Our results demonstrated QUANTITY is an invaluable technique for glycan analysis and bioengineering. 26616285 1 14 theme important 67:75 arg1 class 77:81 an important class 64:81 an important class of macromolecules that play numerous biological functions 64:139 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 1 14 theme important 67:75 arg1 Glycan 54:59 Glycan 54:59 Glycan 54:59 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 2 15 theme Quantitative 142:153 arg1 glycomics--analysis 155:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 4 16 theme biological 680:689 arg1 samples 691:697 multiple biological samples 671:697 multiple biological samples 671:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 5 17 theme knock-in/out 814:825 arg1 enzymes 827:833 knock-in/out enzymes 814:833 knock-in/out enzymes involved in protein glycosylation 814:867 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 2 18 theme structural 326:335 arg1 complexity 337:346 structural complexity 326:346 structural complexity 326:346 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 4 19 theme multiple 671:678 arg1 samples 691:697 multiple biological samples 671:697 multiple biological samples 671:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 0 20 theme Isobaric 13:20 arg1 Tag 22:24 An Isobaric Tag 10:24 QUANTITY: An Isobaric Tag for Quantitative Glycomics. 0:52 QUANTITY: An Isobaric Tag for Quantitative Glycomics. 26616285 3 21 theme global 395:400 arg1 analysis 409:416 global glycan analysis 395:416 global glycan analysis 395:416 As a result, technologies that can facilitate global glycan analysis are highly sought after. 26616285 2 22 theme global 189:194 arg1 level--however 196:209 global level--however 189:209 global level--however 189:209 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 1 23 theme macromolecules 86:99 arg1 class 77:81 an important class 64:81 an important class of macromolecules that play numerous biological functions 64:139 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 1 23 theme macromolecules 86:99 arg1 Glycan 54:59 Glycan 54:59 Glycan 54:59 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 5 24 theme protein 847:853 arg1 glycosylation 855:867 protein glycosylation 847:867 protein glycosylation 847:867 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 5 25 with survey 751:756 arg1 enzymes 827:833 knock-in/out enzymes 814:833 knock-in/out enzymes involved in protein glycosylation 814:867 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 4 26 theme glycomic 648:655 arg1 analysis 657:664 high-throughput glycomic analysis 632:664 high-throughput glycomic analysis from multiple biological samples 632:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 5 27 theme CHO 798:800 arg1 cells 803:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 4 28 theme quantitative 526:537 arg1 approach 539:546 a quantitative approach 524:546 a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples 524:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 28 theme quantitative 526:537 arg1 QUANTITY 460:467 QUANTITY 460:467 QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan) 460:521 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 29 theme Quaternary 470:479 arg1 Tag 507:509 Quaternary Amine Containing Isobaric Tag 470:509 Quaternary Amine Containing Isobaric Tag for Glycan 470:520 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 4 30 theme high-throughput 632:646 arg1 analysis 657:664 high-throughput glycomic analysis 632:664 high-throughput glycomic analysis from multiple biological samples 632:697 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 0 31 theme Quantitative 30:41 arg1 Glycomics 43:51 Quantitative Glycomics 30:51 Quantitative Glycomics 30:51 QUANTITY: An Isobaric Tag for Quantitative Glycomics. 26616285 6 32 dep QUANTITY 895:902 arg1 technique 921:929 an invaluable technique 907:929 an invaluable technique for glycan analysis and bioengineering 907:968 Our results demonstrated QUANTITY is an invaluable technique for glycan analysis and bioengineering. 26616285 2 33 theme glycans 178:184 arg1 glycomics--analysis 155:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis 142:173 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 1 34 theme numerous 111:118 arg1 functions 131:139 numerous biological functions 111:139 numerous biological functions 111:139 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 5 35 theme robust 705:710 arg1 tool 712:715 This robust tool 700:715 This robust tool 700:715 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 2 36 theme technical 259:267 arg1 challenges 269:278 technical challenges 259:278 technical challenges associated with their chemical properties and structural complexity 259:346 Quantitative glycomics--analysis of glycans at global level--however, is far behind genomics and proteomics owing to technical challenges associated with their chemical properties and structural complexity. 26616285 1 37 theme biological 120:129 arg1 functions 131:139 numerous biological functions 111:139 numerous biological functions 111:139 Glycan is an important class of macromolecules that play numerous biological functions. 26616285 4 38 theme mass 598:601 arg1 spectrometry 603:614 mass spectrometry 598:614 mass spectrometry 598:614 Here, we present QUANTITY (Quaternary Amine Containing Isobaric Tag for Glycan), a quantitative approach that can not only enhance detection of glycans by mass spectrometry, but also allow high-throughput glycomic analysis from multiple biological samples. 26616285 5 39 theme bioengineered 761:773 arg1 cells 803:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 5 40 theme Chinese 775:781 arg1 cells 803:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 bioengineered Chinese Hamster Ovary (CHO) cells 761:807 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 26616285 0 41 dep QUANTITY 0:7 arg1 Tag 22:24 An Isobaric Tag 10:24 QUANTITY: An Isobaric Tag for Quantitative Glycomics. 0:52 QUANTITY: An Isobaric Tag for Quantitative Glycomics. 26616285 5 42 theme glycomic 742:749 arg1 survey 751:756 glycomic survey 742:756 glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation 742:867 This robust tool enabled us to accomplish glycomic survey of bioengineered Chinese Hamster Ovary (CHO) cells with knock-in/out enzymes involved in protein glycosylation. 25740940 0 0 theme Haemonchus 70:79 arg1 contortus 81:89 Haemonchus contortus 70:89 Haemonchus contortus 70:89 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 1 1 dep three 241:245 arg1 to 238:239 to 238:239 to 238:239 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 2 theme pole 149:152 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 2 theme pole 149:152 arg1 worm 154:157 barber pole worm 142:157 barber pole worm 142:157 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 6 3 with line 1178:1181 arg1 specificity 1211:1221 the previously defined specificity 1188:1221 the previously defined specificity of nematode glycosyltransferases 1188:1254 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 3 4 from digestions 623:632 arg1 Haemonchus 652:661 Haemonchus 652:661 Haemonchus 652:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 6 5 contain contain 1053:1059 arg2 number 1063:1068 a number 1061:1068 a number of potentially immunogenic glycan epitopes also found in other parasites 1061:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 5 contain contain 1053:1059 arg1 N-glycans 1029:1037 the N-glycans 1025:1037 the N-glycans of Haemonchus 1025:1051 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 6 theme distal 1272:1277 arg1 fucosylation 1279:1290 distal fucosylation 1272:1290 distal fucosylation 1272:1290 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 2 7 with glycans 442:448 arg1 residues 487:494 three hexose residues 474:494 three hexose residues 474:494 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 3 8 theme MALDI-TOF 578:586 arg1 MS/MS 588:592 HPLC and MALDI-TOF MS/MS 569:592 HPLC and MALDI-TOF MS/MS 569:592 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 1 9 theme N'-diacetylchitobiosyl 285:306 arg1 N 283:283 the N 279:283 the N 279:283 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 9 theme N'-diacetylchitobiosyl 285:306 arg1 core 308:311 N'-diacetylchitobiosyl core 285:311 N'-diacetylchitobiosyl core 285:311 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 10 theme distal 380:385 arg1 residue 399:405 the distal core GlcNAc residue 376:405 the distal core GlcNAc residue 376:405 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 5 11 theme antennal 957:964 arg1 fucosylation 966:977 antennal fucosylation 957:977 antennal fucosylation 957:977 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 0 12 theme contortus 81:89 arg1 N-glycans 57:65 N-glycans 57:65 N-glycans of Haemonchus contortus 57:89 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 1 13 theme core 387:390 arg1 residue 399:405 the distal core GlcNAc residue 376:405 the distal core GlcNAc residue 376:405 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 3 14 with combination 597:607 arg1 digestions 623:632 selected digestions 614:632 selected digestions of N-glycans from Haemonchus 614:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 6 15 theme glycan 1097:1102 arg1 epitopes 1104:1111 potentially immunogenic glycan epitopes 1073:1111 potentially immunogenic glycan epitopes also found in other parasites 1073:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 4 16 theme galactosylated 800:813 arg1 α1,3-fucose 822:832 a galactosylated distal α1,3-fucose 798:832 a galactosylated distal α1,3-fucose 798:832 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 17 theme GlcNAc 392:397 arg1 residue 399:405 the distal core GlcNAc residue 376:405 the distal core GlcNAc residue 376:405 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 5 18 theme core 939:942 arg1 α1,6-fucose 944:954 the core α1,6-fucose 935:954 the core α1,6-fucose 935:954 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 6 19 theme nematode 1226:1233 arg1 glycosyltransferases 1235:1254 nematode glycosyltransferases 1226:1254 nematode glycosyltransferases 1226:1254 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 3 20 from Haemonchus 652:661 arg1 N-glycans 637:645 N-glycans 637:645 N-glycans from Haemonchus 637:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 3 20 from Haemonchus 652:661 arg1 digestions 623:632 selected digestions 614:632 selected digestions of N-glycans from Haemonchus 614:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 7 21 theme Haemonchus-type 1448:1462 arg1 N-glycans 1464:1472 Haemonchus-type N-glycans 1448:1472 Haemonchus-type N-glycans 1448:1472 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 6 22 theme epitopes 1104:1111 arg1 number 1063:1068 a number 1061:1068 a number of potentially immunogenic glycan epitopes also found in other parasites 1061:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 5 23 theme α1,6-fucose 944:954 arg1 galactosylation 916:930 galactosylation 916:930 galactosylation of the core α1,6-fucose, antennal fucosylation or modification 916:993 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 6 24 theme glycosyltransferases 1235:1254 arg1 specificity 1211:1221 the previously defined specificity 1188:1221 the previously defined specificity of nematode glycosyltransferases 1188:1254 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 1 25 from contortus 131:139 arg1 first 202:206 first 202:206 first 202:206 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 25 from contortus 131:139 arg1 N-Glycans 92:100 N-Glycans 92:100 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, 92:191 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 5 26 theme Other 890:894 arg1 N-glycans 896:904 Other N-glycans 890:904 Other N-glycans 890:904 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 2 27 contain have 496:499 arg1 glycans 442:448 truncated glycans 432:448 truncated glycans from this organism with three hexose residues 432:494 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 2 27 contain have 496:499 arg2 Man3GlcNAc2Fuc1-3 517:533 the composition Man3GlcNAc2Fuc1-3 501:533 the composition Man3GlcNAc2Fuc1-3 501:533 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 6 28 theme Haemonchus 1042:1051 arg1 N-glycans 1029:1037 the N-glycans 1025:1037 the N-glycans of Haemonchus 1025:1051 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 29 theme α1,6-mannose 1315:1326 arg1 presence 1300:1307 the presence 1296:1307 the presence of an α1,6-mannose 1296:1326 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 29 theme α1,6-mannose 1315:1326 arg1 exclusive 1352:1360 exclusive 1352:1360 exclusive 1352:1360 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 29 theme α1,6-mannose 1315:1326 arg1 fucosylation 1279:1290 distal fucosylation 1272:1290 distal fucosylation 1272:1290 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 4 30 theme distal 815:820 arg1 α1,3-fucose 822:832 a galactosylated distal α1,3-fucose 798:832 a galactosylated distal α1,3-fucose 798:832 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 7 31 theme proteins 1508:1515 arg1 preparation 1481:1491 the preparation 1477:1491 the preparation of recombinant proteins as vaccine candidates 1477:1537 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 4 32 theme Hex3HexNAc2Fuc3 690:704 arg1 glycan 706:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 0 33 theme galactosylated 13:26 arg1 motifs 35:40 galactosylated fucose motifs 13:40 galactosylated fucose motifs 13:40 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 0 34 located present 46:52 arg1 N-glycans 57:65 N-glycans 57:65 N-glycans of Haemonchus contortus 57:89 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 0 34 located present 46:52 arg2 types 4:8 Two types 0:8 Two types of galactosylated fucose motifs 0:40 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 6 35 theme other 1127:1131 arg1 parasites 1133:1141 other parasites 1127:1141 other parasites 1127:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 5 36 theme modification 982:993 arg1 galactosylation 916:930 galactosylation 916:930 galactosylation of the core α1,6-fucose, antennal fucosylation or modification 916:993 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 7 37 theme cell 1416:1419 arg1 lines 1421:1425 cell lines 1416:1425 cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates 1416:1537 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 4 38 theme residues 867:874 arg1 two 849:851 two 849:851 two 849:851 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 4 38 theme residues 867:874 arg1 residues 867:874 the hexose residues 856:874 the hexose residues 856:874 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 39 theme sheep 175:179 arg1 sheep 175:179 sheep 175:179 sheep 175:179 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 39 theme sheep 175:179 arg1 cattle 185:190 cattle 185:190 cattle 185:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 39 theme sheep 175:179 arg1 parasite 163:170 a parasite 161:170 a parasite of sheep and cattle 161:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 39 theme sheep 175:179 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 0 40 theme motifs 35:40 arg1 types 4:8 Two types 0:8 Two types of galactosylated fucose motifs 0:40 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 2 41 theme hexose 480:485 arg1 residues 487:494 three hexose residues 474:494 three hexose residues 474:494 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 4 42 theme core 777:780 arg1 α1,3-fucose 782:792 a proximal core α1,3-fucose 766:792 a proximal core α1,3-fucose 766:792 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 0 43 theme fucose 28:33 arg1 motifs 35:40 galactosylated fucose motifs 13:40 galactosylated fucose motifs 13:40 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 1 44 theme fucose 247:252 arg1 residues 254:261 up to three fucose residues 235:261 up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue 235:405 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 4 45 gly trifucosylated 675:688 arg1 glycan 706:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 46 theme reducing-terminal 331:347 arg1 GlcNAc 358:363 the reducing-terminal proximal GlcNAc 327:363 the reducing-terminal proximal GlcNAc 327:363 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 4 47 theme proximal 768:775 arg1 α1,3-fucose 782:792 a proximal core α1,3-fucose 766:792 a proximal core α1,3-fucose 766:792 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 48 theme cattle 185:190 arg1 sheep 175:179 sheep 175:179 sheep 175:179 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 48 theme cattle 185:190 arg1 cattle 185:190 cattle 185:190 cattle 185:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 48 theme cattle 185:190 arg1 parasite 163:170 a parasite 161:170 a parasite of sheep and cattle 161:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 48 theme cattle 185:190 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 7 49 theme vaccine 1520:1526 arg1 candidates 1528:1537 vaccine candidates 1520:1537 vaccine candidates 1520:1537 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 1 50 theme proximal 349:356 arg1 GlcNAc 358:363 the reducing-terminal proximal GlcNAc 327:363 the reducing-terminal proximal GlcNAc 327:363 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 6 51 theme defined 1203:1209 arg1 specificity 1211:1221 the previously defined specificity 1188:1221 the previously defined specificity of nematode glycosyltransferases 1188:1254 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 4 52 theme hexose 860:865 arg1 residues 867:874 the hexose residues 856:874 the hexose residues 856:874 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 3 53 theme N-glycans 637:645 arg1 digestions 623:632 selected digestions 614:632 selected digestions of N-glycans from Haemonchus 614:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 3 54 theme selected 614:621 arg1 digestions 623:632 selected digestions 614:632 selected digestions of N-glycans from Haemonchus 614:661 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 1 55 theme nematode 111:118 arg1 sheep 175:179 sheep 175:179 sheep 175:179 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 55 theme nematode 111:118 arg1 cattle 185:190 cattle 185:190 cattle 185:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 55 theme nematode 111:118 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 55 theme nematode 111:118 arg1 parasite 163:170 a parasite 161:170 a parasite of sheep and cattle 161:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 55 theme nematode 111:118 arg1 worm 154:157 barber pole worm 142:157 barber pole worm 142:157 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 0 56 attach present 46:52 arg1 N-glycans 57:65 N-glycans 57:65 N-glycans of Haemonchus contortus 57:89 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 0 56 attach present 46:52 arg2 types 4:8 Two types 0:8 Two types of galactosylated fucose motifs 0:40 Two types of galactosylated fucose motifs are present on N-glycans of Haemonchus contortus. 25740940 2 57 theme composition 505:515 arg1 Man3GlcNAc2Fuc1-3 517:533 the composition Man3GlcNAc2Fuc1-3 501:533 the composition Man3GlcNAc2Fuc1-3 501:533 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 6 58 located found 1118:1122 arg1 parasites 1133:1141 other parasites 1127:1141 other parasites 1127:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 58 located found 1118:1122 arg2 epitopes 1104:1111 potentially immunogenic glycan epitopes 1073:1111 potentially immunogenic glycan epitopes also found in other parasites 1073:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 4 59 mod modified 717:724 arg3 α1,6-fucose 740:750 α1,6-fucose 740:750 α1,6-fucose 740:750 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 4 59 mod modified 717:724 arg1 glycan 706:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 4 59 mod modified 717:724 arg3 α1,3-fucose 782:792 a proximal core α1,3-fucose 766:792 a proximal core α1,3-fucose 766:792 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 4 59 mod modified 717:724 arg3 α1,3-fucose 822:832 a galactosylated distal α1,3-fucose 798:832 a galactosylated distal α1,3-fucose 798:832 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 60 theme Haemonchus 120:129 arg1 sheep 175:179 sheep 175:179 sheep 175:179 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 60 theme Haemonchus 120:129 arg1 cattle 185:190 cattle 185:190 cattle 185:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 60 theme Haemonchus 120:129 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 60 theme Haemonchus 120:129 arg1 parasite 163:170 a parasite 161:170 a parasite of sheep and cattle 161:190 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 60 theme Haemonchus 120:129 arg1 worm 154:157 barber pole worm 142:157 barber pole worm 142:157 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 7 61 theme capable 1427:1433 arg1 lines 1421:1425 cell lines 1416:1425 cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates 1416:1537 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 3 62 theme HPLC 569:572 arg1 MS/MS 588:592 HPLC and MALDI-TOF MS/MS 569:592 HPLC and MALDI-TOF MS/MS 569:592 In this study, we have performed HPLC and MALDI-TOF MS/MS in combination with selected digestions of N-glycans from Haemonchus. 25740940 4 63 theme dominant 666:673 arg1 glycan 706:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 4 64 theme trifucosylated 675:688 arg1 glycan 706:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan 664:711 A dominant trifucosylated Hex3HexNAc2Fuc3 glycan was modified not only with α1,6-fucose but also with a proximal core α1,3-fucose and a galactosylated distal α1,3-fucose; thereby, only two of the hexose residues were mannose. 25740940 1 65 from one 369:371 arg1 residue 399:405 the distal core GlcNAc residue 376:405 the distal core GlcNAc residue 376:405 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 2 66 from organism 460:467 arg1 glycans 442:448 truncated glycans 432:448 truncated glycans from this organism with three hexose residues 432:494 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 5 67 theme fucosylation 966:977 arg1 galactosylation 916:930 galactosylation 916:930 galactosylation of the core α1,6-fucose, antennal fucosylation or modification 916:993 Other N-glycans displayed galactosylation of the core α1,6-fucose, antennal fucosylation or modification with phosphorylcholine. 25740940 6 68 theme immunogenic 1085:1095 arg1 epitopes 1104:1111 potentially immunogenic glycan epitopes 1073:1111 potentially immunogenic glycan epitopes also found in other parasites 1073:1141 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 6 69 theme proposed 1151:1158 arg1 structures 1160:1169 our proposed structures 1147:1169 our proposed structures 1147:1169 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 25740940 2 70 theme truncated 432:440 arg1 glycans 442:448 truncated glycans 432:448 truncated glycans from this organism with three hexose residues 432:494 The assumption was that truncated glycans from this organism with three hexose residues have the composition Man3GlcNAc2Fuc1-3. 25740940 7 71 theme recombinant 1496:1506 arg1 proteins 1508:1515 recombinant proteins 1496:1515 recombinant proteins as vaccine candidates 1496:1537 These data are thereby of importance for engineering cell lines capable of mimicking Haemonchus-type N-glycans in the preparation of recombinant proteins as vaccine candidates. 25740940 1 72 theme barber 142:147 arg1 contortus 131:139 the nematode Haemonchus contortus 107:139 the nematode Haemonchus contortus (barber pole worm) 107:158 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 1 72 theme barber 142:147 arg1 worm 154:157 barber pole worm 142:157 barber pole worm 142:157 N-Glycans from the nematode Haemonchus contortus (barber pole worm), a parasite of sheep and cattle, were the first to be described to possess up to three fucose residues associated with the N,N'-diacetylchitobiosyl core, two being on the reducing-terminal proximal GlcNAc and one on the distal core GlcNAc residue. 25740940 6 73 gly fucosylation 1279:1290 arg1 α1,6-mannose 1315:1326 an α1,6-mannose 1312:1326 an α1,6-mannose 1312:1326 Thus, the N-glycans of Haemonchus contain a number of potentially immunogenic glycan epitopes also found in other parasites and our proposed structures are in line with the previously defined specificity of nematode glycosyltransferases as we show that distal fucosylation and the presence of an α1,6-mannose are apparently mutually exclusive. 28853065 1 0 theme targeted 145:152 arg1 delivery 154:161 targeted delivery 145:161 targeted delivery of antitumor drugs 145:180 We studied the possibility of using viscumin lectin (MLI) for targeted delivery of antitumor drugs. 28853065 3 1 from structure 516:524 arg1 general 539:545 general 539:545 general 539:545 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 2 2 gly glycosylation 267:279 arg1 surface 298:304 cell surface 293:304 cell surface 293:304 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 2 3 theme glycosylation 267:279 arg1 profiles 281:288 the glycosylation profiles 263:288 the glycosylation profiles of cell surface in various mouse tissues 263:329 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 3 4 from general 539:545 arg1 structure 516:524 the structure 512:524 the structure of glycan in general 512:545 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 0 5 theme Cell 7:10 arg1 Glycosylation 12:24 Target Cell Glycosylation 0:24 Target Cell Glycosylation 0:24 Target Cell Glycosylation Determines the Biodistribution of Plant Lectin Viscumin. 28853065 3 6 from lectin 476:481 arg1 cells 493:497 tissue cells 486:497 tissue cells 486:497 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 1 7 theme antitumor 166:174 arg1 drugs 176:180 antitumor drugs 166:180 antitumor drugs 166:180 We studied the possibility of using viscumin lectin (MLI) for targeted delivery of antitumor drugs. 28853065 0 8 theme Target 0:5 arg1 Glycosylation 12:24 Target Cell Glycosylation 0:24 Target Cell Glycosylation 0:24 Target Cell Glycosylation Determines the Biodistribution of Plant Lectin Viscumin. 28853065 2 9 theme surface 298:304 arg1 profiles 281:288 the glycosylation profiles 263:288 the glycosylation profiles of cell surface in various mouse tissues 263:329 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 1 10 theme drugs 176:180 arg1 delivery 154:161 targeted delivery 145:161 targeted delivery of antitumor drugs 145:180 We studied the possibility of using viscumin lectin (MLI) for targeted delivery of antitumor drugs. 28853065 3 11 theme glycan 529:534 arg1 structure 516:524 the structure 512:524 the structure of glycan in general 512:545 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 3 12 theme tissue 486:491 arg1 cells 493:497 tissue cells 486:497 tissue cells 486:497 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 3 13 theme only 409:412 arg1 expression 414:423 not only expression 405:423 not only expression of oligosaccharides specifically recognized by the lectin in tissue cells 405:497 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 3 14 theme oligosaccharides 428:443 arg1 expression 414:423 not only expression 405:423 not only expression of oligosaccharides specifically recognized by the lectin in tissue cells 405:497 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 3 15 theme MLI 383:385 arg1 biodistribution 364:378 biodistribution 364:378 biodistribution of MLI 364:385 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 0 16 theme Lectin 66:71 arg1 Viscumin 73:80 Plant Lectin Viscumin 60:80 Plant Lectin Viscumin 60:80 Target Cell Glycosylation Determines the Biodistribution of Plant Lectin Viscumin. 28853065 2 17 theme MLI 195:197 arg1 Affinity 183:190 Affinity 183:190 Affinity of MLI for more than 600 oligosaccharide structures 183:242 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 1 18 theme viscumin 119:126 arg1 lectin 128:133 viscumin lectin 119:133 viscumin lectin (MLI) for targeted delivery of antitumor drugs 119:180 We studied the possibility of using viscumin lectin (MLI) for targeted delivery of antitumor drugs. 28853065 1 18 theme viscumin 119:126 arg1 MLI 136:138 MLI 136:138 MLI 136:138 We studied the possibility of using viscumin lectin (MLI) for targeted delivery of antitumor drugs. 28853065 3 19 from glycan 529:534 arg1 general 539:545 general 539:545 general 539:545 It was found that biodistribution of MLI was determined by not only expression of oligosaccharides specifically recognized by the lectin in tissue cells, but also by the structure of glycan in general. 28853065 2 20 theme mouse 317:321 arg1 tissues 323:329 various mouse tissues 309:329 various mouse tissues 309:329 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 2 21 theme cell 293:296 arg1 surface 298:304 cell surface 293:304 cell surface 293:304 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 2 22 theme various 309:315 arg1 tissues 323:329 various mouse tissues 309:329 various mouse tissues 309:329 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 2 23 from profiles 281:288 arg1 tissues 323:329 various mouse tissues 309:329 various mouse tissues 309:329 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 28853065 0 24 theme Viscumin 73:80 arg1 Biodistribution 41:55 the Biodistribution 37:55 the Biodistribution of Plant Lectin Viscumin 37:80 Target Cell Glycosylation Determines the Biodistribution of Plant Lectin Viscumin. 28853065 0 25 theme Plant 60:64 arg1 Viscumin 73:80 Plant Lectin Viscumin 60:80 Plant Lectin Viscumin 60:80 Target Cell Glycosylation Determines the Biodistribution of Plant Lectin Viscumin. 28853065 2 26 theme oligosaccharide 217:231 arg1 structures 233:242 more than 600 oligosaccharide structures 203:242 more than 600 oligosaccharide structures 203:242 Affinity of MLI for more than 600 oligosaccharide structures was determined and the glycosylation profiles of cell surface in various mouse tissues were analyzed. 24664809 6 0 theme glycans 1169:1175 arg1 RPLC 1132:1135 RPLC 1132:1135 RPLC 1132:1135 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 6 0 theme glycans 1169:1175 arg1 HPAEC 1125:1129 HPAEC 1125:1129 HPAEC 1125:1129 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 6 0 theme glycans 1169:1175 arg1 analyses 1148:1155 HILIC analyses 1142:1155 HILIC analyses 1142:1155 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 3 1 theme peptide 569:575 arg1 bonus 654:658 a bonus 652:658 a bonus 652:658 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 1 theme peptide 569:575 arg1 mapping 577:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 2 theme protein 608:614 arg1 characterization 627:642 protein structural characterization 608:642 protein structural characterization 608:642 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 3 theme data 1378:1381 arg1 analysis 1383:1390 fully automated data analysis 1362:1390 fully automated data analysis 1362:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 3 4 used used 599:602 arg2 mapping 577:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 4 used used 599:602 arg2 bonus 654:658 a bonus 652:658 a bonus 652:658 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 5 with method 1350:1355 arg1 analysis 1383:1390 fully automated data analysis 1362:1390 fully automated data analysis 1362:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 7 6 theme LC-MS/MS 1252:1259 arg1 method 1277:1282 the LC-MS/MS peptide mapping method 1248:1282 the LC-MS/MS peptide mapping method 1248:1282 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 1 7 theme exchange 181:188 arg1 chromatography 190:203 high pH anion exchange chromatography 167:203 high pH anion exchange chromatography (HPAEC) 167:211 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 7 theme exchange 181:188 arg1 HPAEC 206:210 HPAEC 206:210 HPAEC 206:210 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 4 8 theme antibodies 928:937 arg1 digestion 904:912 proteolytic digestion 892:912 proteolytic digestion of monoclonal antibodies (mAbs) 892:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 1 9 attach released 362:369 arg1 glycoprotein 380:391 the glycoprotein 376:391 the glycoprotein 376:391 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 9 attach released 362:369 arg2 glycans 340:346 fluorescence-labeled glycans 319:346 fluorescence-labeled glycans enzymatically released from the glycoprotein 319:391 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 8 10 theme peptide 1334:1340 arg1 method 1494:1499 an alternative method 1479:1499 an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1479:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 10 theme peptide 1334:1340 arg1 method 1350:1355 The LC-MS/MS peptide mapping method 1321:1355 The LC-MS/MS peptide mapping method with fully automated data analysis 1321:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 11 from profiling 1513:1521 arg1 glycoproteins 1572:1584 other glycoproteins 1566:1584 immunoglobulins as well as other glycoproteins with simple N-glycans 1539:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 11 from profiling 1513:1521 arg1 immunoglobulins 1539:1553 immunoglobulins 1539:1553 immunoglobulins as well as other glycoproteins with simple N-glycans 1539:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 1 12 theme reversed 214:221 arg1 chromatography 236:249 reversed phase liquid chromatography 214:249 reversed phase liquid chromatography (RPLC) 214:256 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 12 theme reversed 214:221 arg1 RPLC 252:255 RPLC 252:255 RPLC 252:255 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 4 13 theme developed 764:772 arg1 model 801:805 a recently developed glycopeptide fragmentation model 753:805 a recently developed glycopeptide fragmentation model 753:805 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 3 14 dep used 599:602 arg1 provide 677:683 provide 677:683 can potentially provide glycan profile on each individual glycosylation site 661:736 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 0 15 theme LC-MS/MS 0:7 arg1 mapping 17:23 LC-MS/MS peptide mapping 0:23 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 0:109 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 4 16 theme fragmentation 787:799 arg1 model 801:805 a recently developed glycopeptide fragmentation model 753:805 a recently developed glycopeptide fragmentation model 753:805 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 3 17 theme glycan 685:690 arg1 profile 692:698 glycan profile 685:698 glycan profile 685:698 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 18 theme sample 1406:1411 arg1 preparation 1413:1423 less sample preparation 1401:1423 less sample preparation 1401:1423 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 0 19 theme routine 60:66 arg1 profiling 68:76 routine profiling 60:76 routine profiling of N-glycans in immunoglobulins 60:108 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 1 20 theme fluorescence-labeled 319:338 arg1 glycans 340:346 fluorescence-labeled glycans 319:346 fluorescence-labeled glycans enzymatically released from the glycoprotein 319:391 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 8 21 theme simple 1591:1596 arg1 N-glycans 1598:1606 simple N-glycans 1591:1606 simple N-glycans 1591:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 1 22 theme pH 172:173 arg1 chromatography 190:203 high pH anion exchange chromatography 167:203 high pH anion exchange chromatography (HPAEC) 167:211 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 22 theme pH 172:173 arg1 HPAEC 206:210 HPAEC 206:210 HPAEC 206:210 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 8 23 theme site-specific 1435:1447 arg1 information 1449:1459 site-specific information 1435:1459 site-specific information 1435:1459 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 4 24 from identification 830:843 arg1 digestion 904:912 proteolytic digestion 892:912 proteolytic digestion of monoclonal antibodies (mAbs) 892:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 6 25 theme peptide 1070:1076 arg1 mapping 1078:1084 LC-MS/MS peptide mapping 1061:1084 LC-MS/MS peptide mapping 1061:1084 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 2 26 theme time-consuming 416:429 arg1 preparations 438:449 time-consuming sample preparations 416:449 time-consuming sample preparations 416:449 These methods require time-consuming sample preparations and do not provide site-specific glycosylation information. 24664809 7 27 theme mapping 1269:1275 arg1 method 1277:1282 the LC-MS/MS peptide mapping method 1248:1282 the LC-MS/MS peptide mapping method 1248:1282 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 2 28 theme glycosylation 484:496 arg1 information 498:508 site-specific glycosylation information 470:508 site-specific glycosylation information 470:508 These methods require time-consuming sample preparations and do not provide site-specific glycosylation information. 24664809 4 29 gly N-glycopeptides 871:885 arg2 N-glycopeptides 871:885 N-glycopeptides 871:885 N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs) 871:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 4 29 gly N-glycopeptides 871:885 arg1 digestion 904:912 proteolytic digestion 892:912 proteolytic digestion of monoclonal antibodies (mAbs) 892:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 1 30 theme hydrophilic 262:272 arg1 HILIC 309:313 HILIC 309:313 HILIC 309:313 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 30 theme hydrophilic 262:272 arg1 chromatography 293:306 hydrophilic interaction liquid chromatography 262:306 hydrophilic interaction liquid chromatography (HILIC) 262:314 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 5 31 theme glycoforms 1019:1028 arg1 profiling 1006:1014 accurate profiling 997:1014 accurate profiling of glycoforms 997:1028 Experimental conditions were optimized to achieve accurate profiling of glycoforms. 24664809 4 32 theme N-glycopeptides 871:885 arg1 identification 830:843 automated identification 820:843 automated identification 820:843 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 3 33 theme Liquid 511:516 arg1 LC-MS/MS 559:566 LC-MS/MS 559:566 LC-MS/MS 559:566 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 33 theme Liquid 511:516 arg1 spectrometry 545:556 Liquid chromatography-tandem mass spectrometry 511:556 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 1 34 theme liquid 286:291 arg1 HILIC 309:313 HILIC 309:313 HILIC 309:313 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 34 theme liquid 286:291 arg1 chromatography 293:306 hydrophilic interaction liquid chromatography 262:306 hydrophilic interaction liquid chromatography (HILIC) 262:314 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 35 theme Protein 111:117 arg1 analysis 128:135 Protein N-Glycan analysis 111:135 Protein N-Glycan analysis 111:135 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 3 36 theme individual 708:717 arg1 site 733:736 each individual glycosylation site 703:736 each individual glycosylation site 703:736 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 37 theme alternative 1482:1492 arg1 method 1494:1499 an alternative method 1479:1499 an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1479:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 37 theme alternative 1482:1492 arg1 method 1350:1355 The LC-MS/MS peptide mapping method 1321:1355 The LC-MS/MS peptide mapping method with fully automated data analysis 1321:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 0 38 theme automated 30:38 arg1 processing 45:54 automated data processing 30:54 automated data processing 30:54 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 4 39 theme proteolytic 892:902 arg1 digestion 904:912 proteolytic digestion 892:912 proteolytic digestion of monoclonal antibodies (mAbs) 892:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 0 40 with mapping 17:23 arg1 processing 45:54 automated data processing 30:54 automated data processing 30:54 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 8 41 with immunoglobulins 1539:1553 arg1 N-glycans 1598:1606 simple N-glycans 1591:1606 simple N-glycans 1591:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 1 42 theme phase 223:227 arg1 chromatography 236:249 reversed phase liquid chromatography 214:249 reversed phase liquid chromatography (RPLC) 214:256 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 42 theme phase 223:227 arg1 RPLC 252:255 RPLC 252:255 RPLC 252:255 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 6 43 theme mAb 1189:1191 arg1 molecules 1193:1201 several mAb molecules 1181:1201 several mAb molecules 1181:1201 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 8 44 theme N-glycans 1526:1534 arg1 profiling 1513:1521 routine profiling 1505:1521 routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1505:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 3 45 gly glycosylation 719:731 arg2 site 733:736 each individual glycosylation site 703:736 each individual glycosylation site 703:736 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 7 46 theme glycan 1288:1293 arg1 profiling 1295:1303 glycan profiling 1288:1303 glycan profiling 1288:1303 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 6 47 theme released 1160:1167 arg1 glycans 1169:1175 released glycans 1160:1175 released glycans 1160:1175 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 1 48 theme anion 175:179 arg1 chromatography 190:203 high pH anion exchange chromatography 167:203 high pH anion exchange chromatography (HPAEC) 167:211 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 48 theme anion 175:179 arg1 HPAEC 206:210 HPAEC 206:210 HPAEC 206:210 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 6 49 theme LC-MS/MS 1061:1068 arg1 mapping 1078:1084 LC-MS/MS peptide mapping 1061:1084 LC-MS/MS peptide mapping 1061:1084 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 4 50 theme monoclonal 917:926 arg1 mAbs 940:943 mAbs 940:943 mAbs 940:943 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 4 50 theme monoclonal 917:926 arg1 antibodies 928:937 monoclonal antibodies 917:937 monoclonal antibodies (mAbs) 917:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 8 51 theme LC-MS/MS 1325:1332 arg1 method 1494:1499 an alternative method 1479:1499 an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1479:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 51 theme LC-MS/MS 1325:1332 arg1 method 1350:1355 The LC-MS/MS peptide mapping method 1321:1355 The LC-MS/MS peptide mapping method with fully automated data analysis 1321:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 3 52 theme structural 616:625 arg1 characterization 627:642 protein structural characterization 608:642 protein structural characterization 608:642 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 53 theme mapping 1342:1348 arg1 method 1494:1499 an alternative method 1479:1499 an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1479:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 53 theme mapping 1342:1348 arg1 method 1350:1355 The LC-MS/MS peptide mapping method 1321:1355 The LC-MS/MS peptide mapping method with fully automated data analysis 1321:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 4 54 gly glycopeptide 774:785 arg2 glycopeptide 774:785 a recently developed glycopeptide fragmentation model 753:805 a recently developed glycopeptide fragmentation model 753:805 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 1 55 gly glycoprotein 380:391 arg1 glycoprotein 380:391 the glycoprotein 376:391 the glycoprotein 376:391 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 8 56 gly glycoproteins 1572:1584 arg1 glycoproteins 1572:1584 other glycoproteins 1566:1584 immunoglobulins as well as other glycoproteins with simple N-glycans 1539:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 6 57 theme Glycan 1031:1036 arg1 profiles 1038:1045 Glycan profiles 1031:1045 Glycan profiles obtained from LC-MS/MS peptide mapping 1031:1084 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 0 58 from profiling 68:76 arg1 immunoglobulins 94:108 immunoglobulins 94:108 immunoglobulins 94:108 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 1 59 theme N-Glycan 119:126 arg1 analysis 128:135 Protein N-Glycan analysis 111:135 Protein N-Glycan analysis 111:135 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 8 60 theme other 1566:1570 arg1 glycoproteins 1572:1584 other glycoproteins 1566:1584 immunoglobulins as well as other glycoproteins with simple N-glycans 1539:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 8 61 theme automated 1368:1376 arg1 analysis 1383:1390 fully automated data analysis 1362:1390 fully automated data analysis 1362:1390 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 5 62 theme accurate 997:1004 arg1 profiling 1006:1014 accurate profiling 997:1014 accurate profiling of glycoforms 997:1028 Experimental conditions were optimized to achieve accurate profiling of glycoforms. 24664809 4 63 theme glycopeptide 774:785 arg1 model 801:805 a recently developed glycopeptide fragmentation model 753:805 a recently developed glycopeptide fragmentation model 753:805 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 3 64 theme mass 540:543 arg1 LC-MS/MS 559:566 LC-MS/MS 559:566 LC-MS/MS 559:566 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 64 theme mass 540:543 arg1 spectrometry 545:556 Liquid chromatography-tandem mass spectrometry 511:556 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 5 65 theme Experimental 947:958 arg1 conditions 960:969 Experimental conditions 947:969 Experimental conditions 947:969 Experimental conditions were optimized to achieve accurate profiling of glycoforms. 24664809 0 66 theme N-glycans 81:89 arg1 profiling 68:76 routine profiling 60:76 routine profiling of N-glycans in immunoglobulins 60:108 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 1 67 theme high 167:170 arg1 chromatography 190:203 high pH anion exchange chromatography 167:203 high pH anion exchange chromatography (HPAEC) 167:211 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 67 theme high 167:170 arg1 HPAEC 206:210 HPAEC 206:210 HPAEC 206:210 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 7 68 theme method 1277:1282 arg1 reproducibility 1214:1228 reproducibility 1214:1228 reproducibility 1214:1228 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 7 68 theme method 1277:1282 arg1 linearity 1235:1243 linearity 1235:1243 linearity 1235:1243 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 7 68 theme method 1277:1282 arg1 Accuracy 1204:1211 Accuracy 1204:1211 Accuracy 1204:1211 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 4 69 theme automated 820:828 arg1 identification 830:843 automated identification 820:843 automated identification 820:843 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 7 70 theme peptide 1261:1267 arg1 method 1277:1282 the LC-MS/MS peptide mapping method 1248:1282 the LC-MS/MS peptide mapping method 1248:1282 Accuracy, reproducibility, and linearity of the LC-MS/MS peptide mapping method for glycan profiling were evaluated. 24664809 0 71 theme peptide 9:15 arg1 mapping 17:23 LC-MS/MS peptide mapping 0:23 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 0:109 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 4 72 from digestion 904:912 arg1 identification 830:843 automated identification 820:843 automated identification 820:843 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 4 72 from digestion 904:912 arg1 N-glycopeptides 871:885 N-glycopeptides 871:885 N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs) 871:944 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 24664809 6 73 theme HILIC 1142:1146 arg1 analyses 1148:1155 HILIC analyses 1142:1155 HILIC analyses 1142:1155 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 2 74 theme site-specific 470:482 arg1 information 498:508 site-specific glycosylation information 470:508 site-specific glycosylation information 470:508 These methods require time-consuming sample preparations and do not provide site-specific glycosylation information. 24664809 1 75 theme interaction 274:284 arg1 HILIC 309:313 HILIC 309:313 HILIC 309:313 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 75 theme interaction 274:284 arg1 chromatography 293:306 hydrophilic interaction liquid chromatography 262:306 hydrophilic interaction liquid chromatography (HILIC) 262:314 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 3 76 theme chromatography-tandem 518:538 arg1 LC-MS/MS 559:566 LC-MS/MS 559:566 LC-MS/MS 559:566 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 76 theme chromatography-tandem 518:538 arg1 spectrometry 545:556 Liquid chromatography-tandem mass spectrometry 511:556 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 0 77 theme data 40:43 arg1 processing 45:54 automated data processing 30:54 automated data processing 30:54 LC-MS/MS peptide mapping with automated data processing for routine profiling of N-glycans in immunoglobulins. 24664809 2 78 theme sample 431:436 arg1 preparations 438:449 time-consuming sample preparations 416:449 time-consuming sample preparations 416:449 These methods require time-consuming sample preparations and do not provide site-specific glycosylation information. 24664809 3 79 theme glycosylation 719:731 arg1 site 733:736 each individual glycosylation site 703:736 each individual glycosylation site 703:736 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 8 80 theme routine 1505:1511 arg1 profiling 1513:1521 routine profiling 1505:1521 routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans 1505:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 3 81 theme spectrometry 545:556 arg1 bonus 654:658 a bonus 652:658 a bonus 652:658 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 3 81 theme spectrometry 545:556 arg1 mapping 577:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping 511:583 Liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide mapping is frequently used for protein structural characterization and, as a bonus, can potentially provide glycan profile on each individual glycosylation site. 24664809 1 82 theme liquid 229:234 arg1 chromatography 236:249 reversed phase liquid chromatography 214:249 reversed phase liquid chromatography (RPLC) 214:256 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 1 82 theme liquid 229:234 arg1 RPLC 252:255 RPLC 252:255 RPLC 252:255 Protein N-Glycan analysis is traditionally performed by high pH anion exchange chromatography (HPAEC), reversed phase liquid chromatography (RPLC), or hydrophilic interaction liquid chromatography (HILIC) on fluorescence-labeled glycans enzymatically released from the glycoprotein. 24664809 6 83 theme several 1181:1187 arg1 molecules 1193:1201 several mAb molecules 1181:1201 several mAb molecules 1181:1201 Glycan profiles obtained from LC-MS/MS peptide mapping were compared with those obtained from HPAEC, RPLC, and HILIC analyses of released glycans for several mAb molecules. 24664809 8 84 with glycoproteins 1572:1584 arg1 N-glycans 1598:1606 simple N-glycans 1591:1606 simple N-glycans 1591:1606 The LC-MS/MS peptide mapping method with fully automated data analysis requires less sample preparation, provides site-specific information, and may serve as an alternative method for routine profiling of N-glycans on immunoglobulins as well as other glycoproteins with simple N-glycans. 24664809 4 85 used used 811:814 arg2 model 801:805 a recently developed glycopeptide fragmentation model 753:805 a recently developed glycopeptide fragmentation model 753:805 In this work, a recently developed glycopeptide fragmentation model was used for automated identification, based on their MS/MS, of N-glycopeptides from proteolytic digestion of monoclonal antibodies (mAbs). 25458834 4 0 theme independent 511:521 arg1 interactions 523:534 the glycan-dependent and independent interactions 486:534 the glycan-dependent and independent interactions 486:534 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 5 1 theme noncanonical 683:694 arg1 face 703:706 the noncanonical "side" face 679:706 the noncanonical "side" face of CLEC-2 679:716 Both podoplanin and rhodocytin bind to the noncanonical "side" face of CLEC-2. 25458834 6 2 theme interaction 737:747 arg1 mode 749:752 a common interaction mode 728:752 a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2 728:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 8 3 theme binding 1036:1042 arg1 site 1044:1047 this "second" binding site 1022:1047 this "second" binding site on CLEC-2 1022:1057 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 1 4 theme C-type 178:183 arg1 receptor 197:204 C-type lectin-like receptor 2 178:206 C-type lectin-like receptor 2 (CLEC-2) 178:215 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 1 4 theme C-type 178:183 arg1 CLEC-2 209:214 CLEC-2 209:214 CLEC-2 209:214 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 9 5 theme unique 1064:1069 arg1 modes 1093:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 3 6 dep CLEC-2 385:390 arg1 platelets 407:415 platelets 407:415 platelets 407:415 Rhodocytin, a snake venom, also binds to CLEC-2 and aggregates platelets in a glycan-independent manner. 25458834 8 7 theme subunit 990:996 arg1 terminus 961:968 the C terminus 955:968 the C terminus of the rhodocytin α subunit 955:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 8 8 theme rhodocytin 977:986 arg1 subunit 990:996 the rhodocytin α subunit 973:996 the rhodocytin α subunit 973:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 0 9 theme nonglycosylated 91:105 arg1 rhodocytin 107:116 nonglycosylated rhodocytin 91:116 nonglycosylated rhodocytin 91:116 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 5 10 theme " 701:701 arg1 face 703:706 the noncanonical "side" face 679:706 the noncanonical "side" face of CLEC-2 679:716 Both podoplanin and rhodocytin bind to the noncanonical "side" face of CLEC-2. 25458834 0 11 gly O-glycosylated 61:74 arg1 podoplanin 76:85 O-glycosylated podoplanin 61:85 O-glycosylated podoplanin 61:85 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 6 12 theme common 730:735 arg1 mode 749:752 a common interaction mode 728:752 a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2 728:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 3 13 theme snake 358:362 arg1 venom 364:368 a snake venom 356:368 a snake venom 356:368 Rhodocytin, a snake venom, also binds to CLEC-2 and aggregates platelets in a glycan-independent manner. 25458834 3 13 theme snake 358:362 arg1 Rhodocytin 344:353 Rhodocytin 344:353 Rhodocytin 344:353 Rhodocytin, a snake venom, also binds to CLEC-2 and aggregates platelets in a glycan-independent manner. 25458834 9 14 theme versatile 1075:1083 arg1 modes 1093:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 6 15 theme arginine 818:825 arg1 residues 827:834 the same arginine residues 809:834 the same arginine residues on CLEC-2 809:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 2 16 theme biological 292:301 arg1 processes 303:311 various biological processes 284:311 various biological processes 284:311 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 2 16 theme biological 292:301 arg1 aggregation 331:341 platelet aggregation 322:341 platelet aggregation 322:341 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 9 17 theme binding 1085:1091 arg1 modes 1093:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes 1060:1097 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 1 18 theme lectin-like 185:195 arg1 receptor 197:204 C-type lectin-like receptor 2 178:206 C-type lectin-like receptor 2 (CLEC-2) 178:215 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 1 18 theme lectin-like 185:195 arg1 CLEC-2 209:214 CLEC-2 209:214 CLEC-2 209:214 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 6 19 theme same 813:816 arg1 residues 827:834 the same arginine residues 809:834 the same arginine residues on CLEC-2 809:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 2 20 theme various 284:290 arg1 processes 303:311 various biological processes 284:311 various biological processes 284:311 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 2 20 theme various 284:290 arg1 aggregation 331:341 platelet aggregation 322:341 platelet aggregation 322:341 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 8 21 theme acid 919:922 arg1 residue 924:930 the sialic acid residue 908:930 the sialic acid residue on podoplanin 908:944 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 4 22 theme glycan-dependent 490:505 arg1 interactions 523:534 the glycan-dependent and independent interactions 486:534 the glycan-dependent and independent interactions 486:534 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 0 23 gly nonglycosylated 91:105 arg1 rhodocytin 107:116 nonglycosylated rhodocytin 91:116 nonglycosylated rhodocytin 91:116 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 8 24 theme sialic 912:917 arg1 residue 924:930 the sialic acid residue 908:930 the sialic acid residue on podoplanin 908:944 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 3 25 theme glycan-independent 422:439 arg1 manner 441:446 a glycan-independent manner 420:446 a glycan-independent manner 420:446 Rhodocytin, a snake venom, also binds to CLEC-2 and aggregates platelets in a glycan-independent manner. 25458834 4 26 theme podoplanin 590:599 arg1 studies 579:585 comparative crystallographic studies 550:585 comparative crystallographic studies of podoplanin and rhodocytin 550:614 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 8 27 theme " 1034:1034 arg1 site 1044:1047 this "second" binding site 1022:1047 this "second" binding site on CLEC-2 1022:1057 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 2 28 theme platelet 322:329 arg1 aggregation 331:341 platelet aggregation 322:341 platelet aggregation 322:341 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 0 29 theme C-type 14:19 arg1 CLEC-2 42:47 C-type lectin-like receptor CLEC-2 14:47 C-type lectin-like receptor CLEC-2 14:47 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 4 30 with complex 619:625 arg1 CLEC-2 632:637 CLEC-2 632:637 CLEC-2 632:637 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 8 31 theme second 1028:1033 arg1 site 1044:1047 this "second" binding site 1022:1047 this "second" binding site on CLEC-2 1022:1057 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 8 32 from site 1044:1047 arg1 CLEC-2 1052:1057 CLEC-2 1052:1057 CLEC-2 1052:1057 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 4 33 theme rhodocytin 605:614 arg1 studies 579:585 comparative crystallographic studies 550:585 comparative crystallographic studies of podoplanin and rhodocytin 550:614 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 0 34 theme receptor 33:40 arg1 CLEC-2 42:47 C-type lectin-like receptor CLEC-2 14:47 C-type lectin-like receptor CLEC-2 14:47 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 1 35 gly O-glycoprotein 149:162 arg1 O-glycoprotein 149:162 a transmembrane O-glycoprotein 133:162 a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2) 133:215 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 1 35 gly O-glycoprotein 149:162 arg1 Podoplanin 119:128 Podoplanin 119:128 Podoplanin 119:128 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 8 36 from terminus 961:968 arg1 groups 896:901 Carboxyl groups 887:901 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit 887:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 7 37 theme Other 847:851 arg1 interactions 853:864 Other interactions 847:864 Other interactions 847:864 Other interactions are ligand-specific. 25458834 0 38 theme lectin-like 21:31 arg1 CLEC-2 42:47 C-type lectin-like receptor CLEC-2 14:47 C-type lectin-like receptor CLEC-2 14:47 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 4 39 theme comparative 550:560 arg1 studies 579:585 comparative crystallographic studies 550:585 comparative crystallographic studies of podoplanin and rhodocytin 550:614 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 4 40 theme structural 466:475 arg1 basis 477:481 the structural basis 462:481 the structural basis of the glycan-dependent and independent interactions 462:534 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 9 41 theme functional 1128:1137 arg1 consequences 1139:1150 the functional consequences 1124:1150 the functional consequences of CLEC-2-ligand interactions 1124:1180 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 6 42 from residues 781:788 arg1 ligands 797:803 the ligands 793:803 the ligands 793:803 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 6 42 from residues 781:788 arg1 CLEC-2 839:844 CLEC-2 839:844 CLEC-2 839:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 0 43 theme CLEC-2 42:47 arg1 platform 2:9 A platform 0:9 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 0:117 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 9 44 theme CLEC-2-ligand 1155:1167 arg1 interactions 1169:1180 CLEC-2-ligand interactions 1155:1180 CLEC-2-ligand interactions 1155:1180 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 2 45 theme O-glycan-dependent 222:239 arg1 interaction 241:251 The O-glycan-dependent interaction 218:251 The O-glycan-dependent interaction 218:251 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 5 46 theme side 697:700 arg1 face 703:706 the noncanonical "side" face 679:706 the noncanonical "side" face of CLEC-2 679:716 Both podoplanin and rhodocytin bind to the noncanonical "side" face of CLEC-2. 25458834 4 47 theme crystallographic 562:577 arg1 studies 579:585 comparative crystallographic studies 550:585 comparative crystallographic studies of podoplanin and rhodocytin 550:614 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 5 48 theme CLEC-2 711:716 arg1 face 703:706 the noncanonical "side" face 679:706 the noncanonical "side" face of CLEC-2 679:716 Both podoplanin and rhodocytin bind to the noncanonical "side" face of CLEC-2. 25458834 6 49 theme acidic 774:779 arg1 residues 781:788 consecutive acidic residues 762:788 consecutive acidic residues on the ligands 762:803 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 1 50 theme transmembrane 135:147 arg1 O-glycoprotein 149:162 a transmembrane O-glycoprotein 133:162 a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2) 133:215 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 1 50 theme transmembrane 135:147 arg1 Podoplanin 119:128 Podoplanin 119:128 Podoplanin 119:128 Podoplanin is a transmembrane O-glycoprotein that binds to C-type lectin-like receptor 2 (CLEC-2). 25458834 6 51 from residues 827:834 arg1 ligands 797:803 the ligands 793:803 the ligands 793:803 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 6 51 from residues 827:834 arg1 CLEC-2 839:844 CLEC-2 839:844 CLEC-2 839:844 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 8 52 theme C 959:959 arg1 terminus 961:968 the C terminus 955:968 the C terminus of the rhodocytin α subunit 955:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 6 53 theme consecutive 762:772 arg1 residues 781:788 consecutive acidic residues 762:788 consecutive acidic residues on the ligands 762:803 There is a common interaction mode between consecutive acidic residues on the ligands and the same arginine residues on CLEC-2. 25458834 0 54 theme O-glycosylated 61:74 arg1 podoplanin 76:85 O-glycosylated podoplanin 61:85 O-glycosylated podoplanin 61:85 A platform of C-type lectin-like receptor CLEC-2 for binding O-glycosylated podoplanin and nonglycosylated rhodocytin. 25458834 8 55 theme Carboxyl 887:894 arg1 groups 896:901 Carboxyl groups 887:901 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit 887:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 2 56 theme crucial 267:273 arg1 roles 275:279 crucial roles 267:279 crucial roles 267:279 The O-glycan-dependent interaction seems to play crucial roles in various biological processes, such as platelet aggregation. 25458834 9 57 theme interactions 1169:1180 arg1 consequences 1139:1150 the functional consequences 1124:1150 the functional consequences of CLEC-2-ligand interactions 1124:1180 The unique and versatile binding modes open a way to understand the functional consequences of CLEC-2-ligand interactions. 25458834 4 58 theme interactions 523:534 arg1 basis 477:481 the structural basis 462:481 the structural basis of the glycan-dependent and independent interactions 462:534 To elucidate the structural basis of the glycan-dependent and independent interactions, we performed comparative crystallographic studies of podoplanin and rhodocytin in complex with CLEC-2. 25458834 8 59 from residue 924:930 arg1 podoplanin 935:944 podoplanin 935:944 podoplanin 935:944 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 8 59 from residue 924:930 arg1 groups 896:901 Carboxyl groups 887:901 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit 887:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 25458834 8 60 theme α 988:988 arg1 subunit 990:996 the rhodocytin α subunit 973:996 the rhodocytin α subunit 973:996 Carboxyl groups from the sialic acid residue on podoplanin and from the C terminus of the rhodocytin α subunit interact differently at this "second" binding site on CLEC-2. 27826629 4 0 theme position 507:514 arg1 isomers 488:494 The structural isomers 473:494 The structural isomers of linkage position 473:514 The structural isomers of linkage position are most frequently considered, while stereo isomers are usually assumed from biosynthetic data. 27826629 1 1 theme multiple 133:140 arg1 structures 151:160 multiple isomeric structures 133:160 multiple isomeric structures exposed by sequential mass spectrometry, MS n 133:206 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 4 2 theme biosynthetic 594:605 arg1 data 607:610 biosynthetic data 594:610 biosynthetic data 594:610 The structural isomers of linkage position are most frequently considered, while stereo isomers are usually assumed from biosynthetic data. 27826629 1 3 theme isomeric 142:149 arg1 structures 151:160 multiple isomeric structures 133:160 multiple isomeric structures exposed by sequential mass spectrometry, MS n 133:206 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 6 4 dep LCMS 779:782 arg1 e.g. 773:776 e.g. 773:776 e.g. 773:776 While isomers are usually considered a chromatographic problem (e.g., LCMS, IMMS) and most frequently considered a separations problem, such results will always be challenged by identification and documentation. 27826629 5 5 theme careful 672:678 arg1 attention 680:688 careful attention 672:688 careful attention 672:688 Evaluation of any new sample should be cautious and merits careful attention to empirical data. 27826629 7 6 theme spectral 978:985 arg1 data 987:990 spectral data 978:990 spectral data for characterization (mass and abundance) supported by a universal library match feature 978:1079 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 1 7 theme cancer 82:87 arg1 MDA-MB-231 111:120 MDA-MB-231 111:120 MDA-MB-231 111:120 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 7 theme cancer 82:87 arg1 MCF-7 101:105 MCF-7 101:105 MCF-7 101:105 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 7 theme cancer 82:87 arg1 lines 94:98 two breast cancer cell lines 71:98 two breast cancer cell lines 71:98 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 0 8 theme Isomeric 0:7 arg1 complexity 9:18 Isomeric complexity 0:18 Isomeric complexity of glycosylation 0:35 Isomeric complexity of glycosylation documented by MSn. 27826629 4 9 theme structural 477:486 arg1 isomers 488:494 The structural isomers 473:494 The structural isomers of linkage position 473:514 The structural isomers of linkage position are most frequently considered, while stereo isomers are usually assumed from biosynthetic data. 27826629 5 10 theme sample 635:640 arg1 Evaluation 613:622 Evaluation 613:622 Evaluation of any new sample 613:640 Evaluation of any new sample should be cautious and merits careful attention to empirical data. 27826629 5 11 theme new 631:633 arg1 sample 635:640 any new sample 627:640 any new sample 627:640 Evaluation of any new sample should be cautious and merits careful attention to empirical data. 27826629 3 12 theme varying 444:450 arg1 types 452:456 the varying types 440:456 the varying types 440:456 Probable isomer numbers, when considering both stereo and structural entities, are significant and the varying types are mentioned. 27826629 4 13 theme linkage 499:505 arg1 position 507:514 linkage position 499:514 linkage position 499:514 The structural isomers of linkage position are most frequently considered, while stereo isomers are usually assumed from biosynthetic data. 27826629 7 14 theme direct 940:945 arg1 solution 955:962 a direct spatial solution 938:962 a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature 938:1079 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 1 15 theme breast 75:80 arg1 MDA-MB-231 111:120 MDA-MB-231 111:120 MDA-MB-231 111:120 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 15 theme breast 75:80 arg1 MCF-7 101:105 MCF-7 101:105 MCF-7 101:105 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 15 theme breast 75:80 arg1 lines 94:98 two breast cancer cell lines 71:98 two breast cancer cell lines 71:98 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 7 16 theme spatial 947:953 arg1 solution 955:962 a direct spatial solution 938:962 a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature 938:1079 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 2 17 theme fucosylation 316:327 arg1 structures 329:338 polylactosamine and fucosylation structures 296:338 polylactosamine and fucosylation structures 296:338 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 27826629 7 18 theme universal 1049:1057 arg1 library 1059:1065 a universal library 1047:1065 a universal library match feature 1047:1079 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 3 19 theme structural 399:408 arg1 entities 410:417 structural entities 399:417 structural entities 399:417 Probable isomer numbers, when considering both stereo and structural entities, are significant and the varying types are mentioned. 27826629 1 20 theme cell 89:92 arg1 MDA-MB-231 111:120 MDA-MB-231 111:120 MDA-MB-231 111:120 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 20 theme cell 89:92 arg1 MCF-7 101:105 MCF-7 101:105 MCF-7 101:105 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 20 theme cell 89:92 arg1 lines 94:98 two breast cancer cell lines 71:98 two breast cancer cell lines 71:98 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 21 theme sequential 173:182 arg1 spectrometry 189:200 sequential mass spectrometry 173:200 sequential mass spectrometry 173:200 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 21 theme sequential 173:182 arg1 n 206:206 MS n 203:206 MS n 203:206 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 0 22 theme glycosylation 23:35 arg1 complexity 9:18 Isomeric complexity 0:18 Isomeric complexity of glycosylation 0:35 Isomeric complexity of glycosylation documented by MSn. 27826629 7 23 dep library 1059:1065 arg1 match 1067:1071 match 1067:1071 match feature 1067:1079 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 2 24 theme polylactosamine 296:310 arg1 structures 329:338 polylactosamine and fucosylation structures 296:338 polylactosamine and fucosylation structures 296:338 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 27826629 1 25 theme mass 184:187 arg1 spectrometry 189:200 sequential mass spectrometry 173:200 sequential mass spectrometry 173:200 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 25 theme mass 184:187 arg1 n 206:206 MS n 203:206 MS n 203:206 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 7 26 dep characterization 996:1011 arg1 abundance 1023:1031 abundance 1023:1031 abundance 1023:1031 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 7 26 dep characterization 996:1011 arg1 mass 1014:1017 mass 1014:1017 mass 1014:1017 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 2 27 theme released 218:225 arg1 compositions 234:245 Several released glycan compositions 210:245 Several released glycan compositions 210:245 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 27826629 2 28 theme Several 210:216 arg1 compositions 234:245 Several released glycan compositions 210:245 Several released glycan compositions 210:245 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 27826629 1 29 theme MS 203:204 arg1 spectrometry 189:200 sequential mass spectrometry 173:200 sequential mass spectrometry 173:200 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 1 29 theme MS 203:204 arg1 n 206:206 MS n 203:206 MS n 203:206 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 4 30 theme stereo 554:559 arg1 isomers 561:567 stereo isomers 554:567 stereo isomers 554:567 The structural isomers of linkage position are most frequently considered, while stereo isomers are usually assumed from biosynthetic data. 27826629 5 31 theme empirical 693:701 arg1 data 703:706 empirical data 693:706 empirical data 693:706 Evaluation of any new sample should be cautious and merits careful attention to empirical data. 27826629 6 32 theme such 845:848 arg1 results 850:856 such results 845:856 such results 845:856 While isomers are usually considered a chromatographic problem (e.g., LCMS, IMMS) and most frequently considered a separations problem, such results will always be challenged by identification and documentation. 27826629 7 33 theme MSn 921:923 arg1 data 925:928 MSn data 921:928 MSn data 921:928 MSn data provide a direct spatial solution that includes spectral data for characterization (mass and abundance) supported by a universal library match feature. 27826629 1 34 theme lines 94:98 arg1 Re-analysis 56:66 Re-analysis 56:66 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, 56:121 Re-analysis of two breast cancer cell lines, MCF-7 and MDA-MB-231, has shown multiple isomeric structures exposed by sequential mass spectrometry, MS n . 27826629 2 35 from variations 282:291 arg1 structures 329:338 polylactosamine and fucosylation structures 296:338 polylactosamine and fucosylation structures 296:338 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 27826629 3 36 theme Probable 341:348 arg1 numbers 357:363 Probable isomer numbers 341:363 Probable isomer numbers 341:363 Probable isomer numbers, when considering both stereo and structural entities, are significant and the varying types are mentioned. 27826629 6 37 theme separations 824:834 arg1 problem 836:842 a separations problem 822:842 a separations problem 822:842 While isomers are usually considered a chromatographic problem (e.g., LCMS, IMMS) and most frequently considered a separations problem, such results will always be challenged by identification and documentation. 27826629 6 38 theme chromatographic 748:762 arg1 problem 764:770 a chromatographic problem 746:770 a chromatographic problem (e.g., LCMS, IMMS) 746:789 While isomers are usually considered a chromatographic problem (e.g., LCMS, IMMS) and most frequently considered a separations problem, such results will always be challenged by identification and documentation. 27826629 6 38 theme chromatographic 748:762 arg1 IMMS 785:788 IMMS 785:788 IMMS 785:788 While isomers are usually considered a chromatographic problem (e.g., LCMS, IMMS) and most frequently considered a separations problem, such results will always be challenged by identification and documentation. 27826629 3 39 theme isomer 350:355 arg1 numbers 357:363 Probable isomer numbers 341:363 Probable isomer numbers 341:363 Probable isomer numbers, when considering both stereo and structural entities, are significant and the varying types are mentioned. 27826629 2 40 theme glycan 227:232 arg1 compositions 234:245 Several released glycan compositions 210:245 Several released glycan compositions 210:245 Several released glycan compositions were re-evaluated, which indicated variations in polylactosamine and fucosylation structures. 26859619 3 0 theme Calcium 337:343 arg1 salts 345:349 Calcium salts 337:349 Calcium salts 337:349 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 4 1 theme array 734:738 arg1 structure 702:710 the structure 698:710 the structure of the hydroxyl group array in sucrose 698:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 5 2 theme various 781:787 arg1 analogs 806:812 various monodeoxysucrose analogs 781:812 various monodeoxysucrose analogs 781:812 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 4 3 theme group 728:732 arg1 array 734:738 the hydroxyl group array 715:738 the hydroxyl group array in sucrose 715:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 0 4 theme Ion 101:103 arg1 Presence 81:88 the Presence 77:88 the Presence of Calcium Ion and Trimethylamine 77:122 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 3 5 theme high 403:406 arg1 site-selectivity 408:423 high site-selectivity 403:423 high site-selectivity 403:423 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 0 6 theme Calcium 93:99 arg1 Ion 101:103 Calcium Ion 93:103 Calcium Ion 93:103 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 4 7 theme nonenzymatic 505:516 arg1 syntheses 542:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 5 8 theme bond-forming 910:921 arg1 reaction 923:930 this aqueous glycosyl bond-forming reaction 888:930 this aqueous glycosyl bond-forming reaction 888:930 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 3 9 theme tertiary 357:364 arg1 base 372:375 a tertiary amine base 355:375 a tertiary amine base 355:375 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 4 10 from structure 702:710 arg1 sucrose 743:749 sucrose 743:749 sucrose 743:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 1 11 theme sucrosyl 178:185 arg1 acceptors 187:195 sucrosyl acceptors 178:195 sucrosyl acceptors 178:195 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 3 12 theme amine 366:370 arg1 base 372:375 a tertiary amine base 355:375 a tertiary amine base 355:375 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 4 13 theme oligosaccharide 526:540 arg1 syntheses 542:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 2 14 theme room 289:292 arg1 temperature 294:304 room temperature 289:304 room temperature 289:304 This reaction proceeds at room temperature in an aqueous solvent mixture. 26859619 0 15 theme Trimethylamine 109:122 arg1 Presence 81:88 the Presence 77:88 the Presence of Calcium Ion and Trimethylamine 77:122 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 5 16 theme hydroxyl 854:861 arg1 groups 863:868 the hydroxyl groups 850:868 the hydroxyl groups 850:868 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 4 17 theme aqueous 518:524 arg1 syntheses 542:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 nonenzymatic aqueous oligosaccharide syntheses 505:550 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 0 18 theme Aqueous 0:6 arg1 Glycosylation 8:20 Aqueous Glycosylation 0:20 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl 0:62 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 5 19 from time 969:972 arg1 reaction 923:930 this aqueous glycosyl bond-forming reaction 888:930 this aqueous glycosyl bond-forming reaction 888:930 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 5 19 from time 969:972 arg1 site-selectivity 940:955 the site-selectivity 936:955 the site-selectivity at the same time 936:972 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 3 20 with reaction 389:396 arg1 site-selectivity 408:423 high site-selectivity 403:423 high site-selectivity 403:423 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 5 21 theme solution 756:763 arg1 conformation 765:776 The solution conformation 752:776 The solution conformation of various monodeoxysucrose analogs 752:812 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 1 22 theme glycosyl 201:208 arg1 donors 219:224 glycosyl fluoride donors 201:224 glycosyl fluoride donors 201:224 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 0 23 theme Unprotected 25:35 arg1 Sucrose 37:43 Unprotected Sucrose 25:43 Unprotected Sucrose Employing Glycosyl 25:62 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 5 24 theme groups 863:868 arg1 nature 840:845 the co-operative nature 823:845 the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time 823:972 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 1 25 link derived 239:245 arg1 trisaccharides 247:260 the derived trisaccharides 235:260 the derived trisaccharides 235:260 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 2 26 theme solvent 320:326 arg1 mixture 328:334 an aqueous solvent mixture 309:334 an aqueous solvent mixture 309:334 This reaction proceeds at room temperature in an aqueous solvent mixture. 26859619 1 27 theme fluoride 210:217 arg1 donors 219:224 glycosyl fluoride donors 201:224 glycosyl fluoride donors 201:224 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 2 28 theme aqueous 312:318 arg1 mixture 328:334 an aqueous solvent mixture 309:334 an aqueous solvent mixture 309:334 This reaction proceeds at room temperature in an aqueous solvent mixture. 26859619 4 29 theme hydroxyl 719:726 arg1 group 728:732 the hydroxyl group 715:732 the hydroxyl group array in sucrose 715:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 5 30 theme co-operative 827:838 arg1 nature 840:845 the co-operative nature 823:845 the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time 823:972 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 3 31 dep 3'-position 440:450 arg1 the 436:438 the 436:438 the 436:438 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 5 32 theme aqueous 893:899 arg1 reaction 923:930 this aqueous glycosyl bond-forming reaction 888:930 this aqueous glycosyl bond-forming reaction 888:930 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 0 33 theme Sucrose 37:43 arg1 Glycosylation 8:20 Aqueous Glycosylation 0:20 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl 0:62 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 0 34 gly Glycosylation 8:20 arg1 Sucrose 37:43 Unprotected Sucrose 25:43 Unprotected Sucrose Employing Glycosyl 25:62 Aqueous Glycosylation of Unprotected Sucrose Employing Glycosyl Fluorides in the Presence of Calcium Ion and Trimethylamine. 26859619 3 35 theme fructofuranoside 474:489 arg1 unit 491:494 the fructofuranoside unit 470:494 the fructofuranoside unit 470:494 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 4 36 theme mechanistic 572:582 arg1 studies 584:590 mechanistic studies 572:590 mechanistic studies 572:590 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 4 37 dep hypothesized 670:681 arg1 related 687:693 related 687:693 related 687:693 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 5 38 theme same 964:967 arg1 time 969:972 the same time 960:972 the same time 960:972 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 4 39 theme selectivity 648:658 arg1 origin 634:639 the origin 630:639 the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose 630:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 4 40 from sucrose 743:749 arg1 structure 702:710 the structure 698:710 the structure of the hydroxyl group array in sucrose 698:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 1 41 theme derived 239:245 arg1 trisaccharides 247:260 the derived trisaccharides 235:260 the derived trisaccharides 235:260 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 5 42 theme monodeoxysucrose 789:804 arg1 analogs 806:812 various monodeoxysucrose analogs 781:812 various monodeoxysucrose analogs 781:812 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 5 43 theme glycosyl 901:908 arg1 reaction 923:930 this aqueous glycosyl bond-forming reaction 888:930 this aqueous glycosyl bond-forming reaction 888:930 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 3 44 theme unit 491:494 arg1 1'-position 455:465 1'-position 455:465 1'-position 455:465 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 3 44 theme unit 491:494 arg1 3'-position 440:450 3'-position 440:450 3'-position 440:450 Calcium salts and a tertiary amine base promote the reaction with high site-selectivity for either the 3'-position or 1'-position of the fructofuranoside unit. 26859619 1 45 theme synthetic 137:145 arg1 reaction 161:168 a synthetic glycosylation reaction 135:168 a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides 135:260 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 5 46 theme analogs 806:812 arg1 conformation 765:776 The solution conformation 752:776 The solution conformation of various monodeoxysucrose analogs 752:812 The solution conformation of various monodeoxysucrose analogs revealed the co-operative nature of the hydroxyl groups in mediating both this aqueous glycosyl bond-forming reaction and the site-selectivity at the same time. 26859619 4 47 dep carried 597:603 arg1 out 605:607 out 605:607 out 605:607 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 26859619 1 48 theme glycosylation 147:159 arg1 reaction 161:168 a synthetic glycosylation reaction 135:168 a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides 135:260 We report a synthetic glycosylation reaction between sucrosyl acceptors and glycosyl fluoride donors to yield the derived trisaccharides. 26859619 4 49 from array 734:738 arg1 sucrose 743:749 sucrose 743:749 sucrose 743:749 Because nonenzymatic aqueous oligosaccharide syntheses are underdeveloped, mechanistic studies were carried out in order to identify the origin of the selectivity, which we hypothesized was related to the structure of the hydroxyl group array in sucrose. 27060996 8 0 theme N-acetyl 1371:1378 arg1 donors 1402:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 1 1 theme α-sialosides 144:155 arg1 reactions 186:194 the most difficult reactions 167:194 the most difficult reactions in carbohydrate chemistry 167:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 1 theme α-sialosides 144:155 arg1 one 160:162 one 160:162 one 160:162 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 1 theme α-sialosides 144:155 arg1 process 298:304 both a thermodynamically and kinetically disfavored process 246:304 process 298:304 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 1 theme α-sialosides 144:155 arg1 synthesis 131:139 The synthesis 127:139 The synthesis of α-sialosides 127:155 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 8 2 contain containing 1296:1305 arg1 disialosides 1283:1294 α(2,9) disialosides 1276:1294 α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups 1276:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 8 2 contain containing 1296:1305 arg2 groups 1334:1339 N-acetyl and/or N-glycolyl groups 1307:1339 N-acetyl and/or N-glycolyl groups 1307:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 0 3 from α-Glycosidation 4:18 arg1 Absence 90:96 the Absence 86:96 the Absence of a Nitrile Solvent Effect 86:124 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 6 4 theme O-acetyl 865:872 arg1 protection 874:883 the O-acetyl protection 861:883 the O-acetyl protection of the sialyl donor 861:903 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 6 4 theme O-acetyl 865:872 arg1 factor 917:922 a major factor 909:922 a major factor 909:922 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 7 5 theme protecting 990:999 arg1 groups 1023:1028 the hydroxy groups 1011:1028 the hydroxy groups at the 4,7,8 position on the sialyl donor 1011:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 7 5 theme protecting 990:999 arg1 groups 1001:1006 the protecting groups 986:1006 the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor 986:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 3 6 theme unprotected 491:501 arg1 donors 527:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 7 7 gly sialylation 1151:1161 arg1 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside 1091:1139 the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside 1087:1139 the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside 1087:1139 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 7 8 theme groups 1001:1006 arg1 tuning 976:981 tuning 976:981 tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor 976:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 3 9 theme N-acetyl 503:510 arg1 donors 527:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 8 10 theme sialyl 1395:1400 arg1 donors 1402:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 4 11 theme CH2 642:644 arg1 Cl2 646:648 CH2 Cl2 642:648 CH2 Cl2 with a good α-selectivity 642:674 The 9-O-benzyl-N-acetylthiosialoside underwent glycosidation in CH2 Cl2 with a good α-selectivity. 27060996 7 12 theme excellent 1168:1176 arg1 α-selectivity 1178:1190 excellent α-selectivity 1168:1190 excellent α-selectivity in CH2 Cl2 1168:1201 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 8 13 theme N-glycolyl 1384:1393 arg1 donors 1402:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 the two N-acetyl and N-glycolyl sialyl donors 1363:1407 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 0 14 theme Nitrile 103:109 arg1 Effect 119:124 a Nitrile Solvent Effect 101:124 a Nitrile Solvent Effect 101:124 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 2 15 theme N-acetyl 406:413 arg1 acids 422:426 N-acetyl sialic acids 406:426 N-acetyl sialic acids 406:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 3 16 theme N-glycolyl 516:525 arg1 donors 527:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 partially unprotected N-acetyl and N-glycolyl donors 481:532 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 2 17 theme α-selective 377:387 arg1 glycosidation 389:401 the α-selective glycosidation 373:401 the α-selective glycosidation of N-acetyl sialic acids 373:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 1 18 dep process 298:304 arg1 both 246:249 both 246:249 both 246:249 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 5 19 theme same 809:812 arg1 conditions 823:832 the same reaction conditions 805:832 the same reaction conditions 805:832 On the other hand, the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside was converted to β-sialoside as a major product under the same reaction conditions. 27060996 3 20 theme donors 527:532 arg1 α-glycosidation 462:476 the α-glycosidation 458:476 the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect 458:575 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 4 21 theme good 657:660 arg1 α-selectivity 662:674 a good α-selectivity 655:674 a good α-selectivity 655:674 The 9-O-benzyl-N-acetylthiosialoside underwent glycosidation in CH2 Cl2 with a good α-selectivity. 27060996 8 22 theme method 1239:1244 arg1 utility 1224:1230 the utility 1220:1230 the utility of the method 1220:1244 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 5 23 theme major 785:789 arg1 product 791:797 a major product 783:797 a major product under the same reaction conditions 783:832 On the other hand, the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside was converted to β-sialoside as a major product under the same reaction conditions. 27060996 5 23 theme major 785:789 arg1 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside 700:749 the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside 696:749 the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside 696:749 On the other hand, the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside was converted to β-sialoside as a major product under the same reaction conditions. 27060996 3 24 from α-glycosidation 462:476 arg1 absence 541:547 the absence 537:547 the absence of a nitrile solvent effect 537:575 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 3 25 theme nitrile 554:560 arg1 effect 570:575 a nitrile solvent effect 552:575 a nitrile solvent effect 552:575 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 8 26 theme N-acetyl 1307:1314 arg1 groups 1334:1339 N-acetyl and/or N-glycolyl groups 1307:1339 N-acetyl and/or N-glycolyl groups 1307:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 8 27 theme disialosides 1283:1294 arg1 synthesis 1263:1271 straightforward synthesis 1247:1271 straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups 1247:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 8 28 theme straightforward 1247:1261 arg1 synthesis 1263:1271 straightforward synthesis 1247:1271 straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups 1247:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 0 29 theme Effect 119:124 arg1 Absence 90:96 the Absence 86:96 the Absence of a Nitrile Solvent Effect 86:124 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 7 30 theme groups 1023:1028 arg1 groups 1023:1028 the hydroxy groups 1011:1028 the hydroxy groups at the 4,7,8 position on the sialyl donor 1011:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 7 30 theme groups 1023:1028 arg1 groups 1001:1006 the protecting groups 986:1006 the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor 986:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 2 31 theme sialic 415:420 arg1 acids 422:426 N-acetyl sialic acids 406:426 N-acetyl sialic acids 406:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 2 32 theme acids 422:426 arg1 glycosidation 389:401 the α-selective glycosidation 373:401 the α-selective glycosidation of N-acetyl sialic acids 373:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 0 33 theme Solvent 111:117 arg1 Effect 119:124 a Nitrile Solvent Effect 101:124 a Nitrile Solvent Effect 101:124 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 7 34 from groups 1001:1006 arg1 position 1043:1050 the 4,7,8 position 1033:1050 the 4,7,8 position on the sialyl donor 1033:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 1 35 theme reactions 186:194 arg1 reactions 186:194 the most difficult reactions 167:194 the most difficult reactions in carbohydrate chemistry 167:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 35 theme reactions 186:194 arg1 one 160:162 one 160:162 one 160:162 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 35 theme reactions 186:194 arg1 process 298:304 both a thermodynamically and kinetically disfavored process 246:304 process 298:304 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 35 theme reactions 186:194 arg1 synthesis 131:139 The synthesis 127:139 The synthesis of α-sialosides 127:155 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 0 36 theme N-Acetyl 45:52 arg1 Donors 76:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 1 37 theme disfavored 287:296 arg1 reactions 186:194 the most difficult reactions 167:194 the most difficult reactions in carbohydrate chemistry 167:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 37 theme disfavored 287:296 arg1 one 160:162 one 160:162 one 160:162 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 37 theme disfavored 287:296 arg1 synthesis 131:139 The synthesis 127:139 The synthesis of α-sialosides 127:155 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 1 37 theme disfavored 287:296 arg1 process 298:304 both a thermodynamically and kinetically disfavored process 246:304 process 298:304 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 6 38 theme donor 899:903 arg1 protection 874:883 the O-acetyl protection 861:883 the O-acetyl protection of the sialyl donor 861:903 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 6 38 theme donor 899:903 arg1 factor 917:922 a major factor 909:922 a major factor 909:922 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 7 39 theme CH2 1195:1197 arg1 Cl2 1199:1201 CH2 Cl2 1195:1201 CH2 Cl2 1195:1201 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 0 40 theme Unprotected 33:43 arg1 Donors 76:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 5 41 theme reaction 814:821 arg1 conditions 823:832 the same reaction conditions 805:832 the same reaction conditions 805:832 On the other hand, the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside was converted to β-sialoside as a major product under the same reaction conditions. 27060996 6 42 theme sialyl 892:897 arg1 donor 899:903 the sialyl donor 888:903 the sialyl donor 888:903 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 7 43 theme 4,7,8 1037:1041 arg1 position 1043:1050 the 4,7,8 position 1033:1050 the 4,7,8 position on the sialyl donor 1033:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 1 44 theme carbohydrate 199:210 arg1 chemistry 212:220 carbohydrate chemistry 199:220 carbohydrate chemistry 199:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 0 45 theme N-Glycolyl 58:67 arg1 Donors 76:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 7 46 theme sialyl 1059:1064 arg1 donor 1066:1070 the sialyl donor 1055:1070 the sialyl donor 1055:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 3 47 theme solvent 562:568 arg1 effect 570:575 a nitrile solvent effect 552:575 a nitrile solvent effect 552:575 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 6 48 theme major 911:915 arg1 factor 917:922 a major factor 909:922 a major factor 909:922 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 6 48 theme major 911:915 arg1 protection 874:883 the O-acetyl protection 861:883 the O-acetyl protection of the sialyl donor 861:903 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 8 49 theme α 1276:1276 arg1 disialosides 1283:1294 α(2,9) disialosides 1276:1294 α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups 1276:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 7 50 from α-selectivity 1178:1190 arg1 Cl2 1199:1201 CH2 Cl2 1195:1201 CH2 Cl2 1195:1201 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 5 51 theme other 684:688 arg1 hand 690:693 the other hand 680:693 the other hand 680:693 On the other hand, the 4,7,8-O-triacetyl-9-O-benzyl-N-acetylthiosialoside was converted to β-sialoside as a major product under the same reaction conditions. 27060996 3 52 theme effect 570:575 arg1 absence 541:547 the absence 537:547 the absence of a nitrile solvent effect 537:575 In this report, we report on the α-glycosidation of partially unprotected N-acetyl and N-glycolyl donors in the absence of a nitrile solvent effect. 27060996 6 53 theme sialylation 957:967 arg1 α-selectivity 940:952 the α-selectivity 936:952 the α-selectivity of sialylation 936:967 The results indicate that the O-acetyl protection of the sialyl donor was a major factor in reducing the α-selectivity of sialylation. 27060996 1 54 from reactions 186:194 arg1 chemistry 212:220 carbohydrate chemistry 199:220 carbohydrate chemistry 199:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 2 55 theme acetonitrile 318:329 arg1 solution 360:367 an effective solution 347:367 an effective solution for the α-selective glycosidation of N-acetyl sialic acids 347:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 2 55 theme acetonitrile 318:329 arg1 use 311:313 The use 307:313 The use of acetonitrile as a solvent 307:342 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 8 56 theme N-glycolyl 1323:1332 arg1 groups 1334:1339 N-acetyl and/or N-glycolyl groups 1307:1339 N-acetyl and/or N-glycolyl groups 1307:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 4 57 with Cl2 646:648 arg1 α-selectivity 662:674 a good α-selectivity 655:674 a good α-selectivity 655:674 The 9-O-benzyl-N-acetylthiosialoside underwent glycosidation in CH2 Cl2 with a good α-selectivity. 27060996 7 58 theme hydroxy 1015:1021 arg1 groups 1023:1028 the hydroxy groups 1011:1028 the hydroxy groups at the 4,7,8 position on the sialyl donor 1011:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 0 59 theme Donors 76:81 arg1 α-Glycosidation 4:18 The α-Glycosidation 0:18 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 0:125 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 27060996 7 60 from position 1043:1050 arg1 groups 1023:1028 the hydroxy groups 1011:1028 the hydroxy groups at the 4,7,8 position on the sialyl donor 1011:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 7 60 from position 1043:1050 arg1 donor 1066:1070 the sialyl donor 1055:1070 the sialyl donor 1055:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 7 60 from position 1043:1050 arg1 groups 1001:1006 the protecting groups 986:1006 the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor 986:1070 After tuning of the protecting groups of the hydroxy groups at the 4,7,8 position on the sialyl donor, we found that the 9-O-benzyl-4-O-chloroacetyl-N-acetylthiosialoside underwent sialylation with excellent α-selectivity in CH2 Cl2 . 27060996 1 61 theme difficult 176:184 arg1 reactions 186:194 the most difficult reactions 167:194 the most difficult reactions in carbohydrate chemistry 167:220 The synthesis of α-sialosides is one of the most difficult reactions in carbohydrate chemistry and is considered to be both a thermodynamically and kinetically disfavored process. 27060996 2 62 theme effective 350:358 arg1 solution 360:367 an effective solution 347:367 an effective solution for the α-selective glycosidation of N-acetyl sialic acids 347:426 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 2 62 theme effective 350:358 arg1 use 311:313 The use 307:313 The use of acetonitrile as a solvent 307:342 The use of acetonitrile as a solvent is an effective solution for the α-selective glycosidation of N-acetyl sialic acids. 27060996 8 63 theme 2,9 1278:1280 arg1 disialosides 1283:1294 α(2,9) disialosides 1276:1294 α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups 1276:1339 To demonstrate the utility of the method, straightforward synthesis of α(2,9) disialosides containing N-acetyl and/or N-glycolyl groups was achieved by using the two N-acetyl and N-glycolyl sialyl donors. 27060996 0 64 theme Sialyl 69:74 arg1 Donors 76:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors 23:81 The α-Glycosidation of Partially Unprotected N-Acetyl and N-Glycolyl Sialyl Donors in the Absence of a Nitrile Solvent Effect. 29079775 4 0 theme unprotected 741:751 arg1 groups 762:767 one to three unprotected hydroxyl groups 728:767 one to three unprotected hydroxyl groups 728:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 1 1 from study 92:96 arg1 systems 141:147 biological systems 130:147 biological systems 130:147 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 2 2 theme glycosylation 388:400 arg1 methods 402:408 stereoselective glycosylation methods 372:408 stereoselective glycosylation methods 372:408 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 3 3 theme protection/deprotection 565:587 arg1 steps 589:593 protection/deprotection steps 565:593 protection/deprotection steps required for further elongation 565:625 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 3 4 theme further 608:614 arg1 elongation 616:625 further elongation 608:625 further elongation 608:625 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 4 5 theme desired 823:829 arg1 products 831:838 the desired products 819:838 the desired products 819:838 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 2 6 theme significant 307:317 arg1 breakthroughs 319:331 significant breakthroughs 307:331 significant breakthroughs in recent years 307:347 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 4 7 theme various 781:787 arg1 acceptors 798:806 various glycosyl acceptors 781:806 various glycosyl acceptors 781:806 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 1 8 theme biological 130:139 arg1 systems 141:147 biological systems 130:147 biological systems 130:147 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 4 9 theme wide 689:692 arg1 array 694:698 a wide array 687:698 a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups 687:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 1 10 theme Mechanistic 80:90 arg1 study 92:96 Mechanistic study 80:96 Mechanistic study of carbohydrate interactions in biological systems 80:147 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 2 11 from stereo-configuration 233:252 arg1 branching 296:304 branching 296:304 branching 296:304 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 12 theme chains 439:444 arg1 methods 402:408 stereoselective glycosylation methods 372:408 stereoselective glycosylation methods 372:408 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 12 theme chains 439:444 arg1 assembly 420:427 facile assembly 413:427 facile assembly of glycan chains 413:444 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 4 13 theme in 662:663 arg1 reagent 678:684 an in situ masking reagent 659:684 an in situ masking reagent 659:684 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 13 theme in 662:663 arg1 triflate 647:654 dialkylboryl triflate 634:654 dialkylboryl triflate 634:654 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 5 14 theme complex 1010:1016 arg1 synthesis 1033:1041 complex glycoconjugate synthesis 1010:1041 complex glycoconjugate synthesis 1010:1041 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 4 15 theme masking 670:676 arg1 reagent 678:684 an in situ masking reagent 659:684 an in situ masking reagent 659:684 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 15 theme masking 670:676 arg1 triflate 647:654 dialkylboryl triflate 634:654 dialkylboryl triflate 634:654 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 2 16 theme glycan 432:437 arg1 chains 439:444 glycan chains 432:444 glycan chains 432:444 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 0 17 theme minimalist 2:11 arg1 approach 13:20 A minimalist approach 0:20 A minimalist approach to stereoselective glycosylation with unprotected donors 0:77 A minimalist approach to stereoselective glycosylation with unprotected donors. 29079775 3 18 theme steps 589:593 arg1 formation 540:548 stereoselective glycosidic bond formation 508:548 stereoselective glycosidic bond formation 508:548 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 3 18 theme steps 589:593 arg1 removal 554:560 removal 554:560 removal of protection/deprotection steps required for further elongation 554:625 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 4 19 theme donors 712:717 arg1 array 694:698 a wide array 687:698 a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups 687:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 1 20 theme structures 199:208 arg1 synthesis 172:180 the chemical synthesis 159:180 the chemical synthesis of these complex structures 159:208 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 5 21 theme straightforward 949:963 arg1 access 965:970 straightforward access 949:970 straightforward access 949:970 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 1 22 theme carbohydrate 101:112 arg1 interactions 114:125 carbohydrate interactions 101:125 carbohydrate interactions 101:125 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 5 23 theme important 975:983 arg1 scaffolds 996:1004 important structural scaffolds 975:1004 important structural scaffolds for complex glycoconjugate synthesis 975:1041 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 2 24 theme recent 336:341 arg1 years 343:347 recent years 336:347 recent years 336:347 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 25 from linkage 271:277 arg1 stereo-configuration 233:252 the specific stereo-configuration 220:252 the specific stereo-configuration at each anomeric linkage 220:277 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 25 from linkage 271:277 arg1 diversity 283:291 diversity 283:291 diversity in branching 283:304 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 4 26 theme good 845:848 arg1 control 850:856 good control 845:856 good control over regioselectivity and stereoselectivity 845:900 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 27 theme hydroxyl 753:760 arg1 groups 762:767 one to three unprotected hydroxyl groups 728:767 one to three unprotected hydroxyl groups 728:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 3 28 theme stereoselective 508:522 arg1 formation 540:548 stereoselective glycosidic bond formation 508:548 stereoselective glycosidic bond formation 508:548 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 4 29 theme dialkylboryl 634:645 arg1 reagent 678:684 an in situ masking reagent 659:684 an in situ masking reagent 659:684 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 29 theme dialkylboryl 634:645 arg1 triflate 647:654 dialkylboryl triflate 634:654 dialkylboryl triflate 634:654 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 30 dep in 662:663 arg1 situ 665:668 situ 665:668 situ 665:668 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 2 31 theme anomeric 262:269 arg1 linkage 271:277 each anomeric linkage 257:277 each anomeric linkage 257:277 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 32 from diversity 283:291 arg1 branching 296:304 branching 296:304 branching 296:304 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 4 33 theme glycosyl 703:710 arg1 donors 712:717 glycosyl donors 703:717 glycosyl donors carrying one to three unprotected hydroxyl groups 703:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 0 34 theme stereoselective 25:39 arg1 glycosylation 41:53 stereoselective glycosylation 25:53 stereoselective glycosylation with unprotected donors 25:77 A minimalist approach to stereoselective glycosylation with unprotected donors. 29079775 5 35 theme access 965:970 arg1 feasibility 934:944 the feasibility 930:944 the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis 930:1041 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 0 36 theme unprotected 60:70 arg1 donors 72:77 unprotected donors 60:77 unprotected donors 60:77 A minimalist approach to stereoselective glycosylation with unprotected donors. 29079775 5 37 theme structural 985:994 arg1 scaffolds 996:1004 important structural scaffolds 975:1004 important structural scaffolds for complex glycoconjugate synthesis 975:1041 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 3 38 theme unification 470:480 arg1 approach 482:489 the unification approach 466:489 the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation 466:625 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 2 39 theme stereoselective 372:386 arg1 methods 402:408 stereoselective glycosylation methods 372:408 stereoselective glycosylation methods 372:408 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 2 40 theme specific 224:231 arg1 stereo-configuration 233:252 the specific stereo-configuration 220:252 the specific stereo-configuration at each anomeric linkage 220:277 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 3 41 theme glycosidic 524:533 arg1 formation 540:548 stereoselective glycosidic bond formation 508:548 stereoselective glycosidic bond formation 508:548 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 1 42 theme interactions 114:125 arg1 study 92:96 Mechanistic study 80:96 Mechanistic study of carbohydrate interactions in biological systems 80:147 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 4 43 dep three 735:739 arg1 to 732:733 to 732:733 to 732:733 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 5 44 theme glycoconjugate 1018:1031 arg1 synthesis 1033:1041 complex glycoconjugate synthesis 1010:1041 complex glycoconjugate synthesis 1010:1041 This approach demonstrates the feasibility of straightforward access to important structural scaffolds for complex glycoconjugate synthesis. 29079775 3 45 theme bond 535:538 arg1 formation 540:548 stereoselective glycosidic bond formation 508:548 stereoselective glycosidic bond formation 508:548 Here, we introduce the unification approach that offers both stereoselective glycosidic bond formation and removal of protection/deprotection steps required for further elongation. 29079775 2 46 from breakthroughs 319:331 arg1 years 343:347 recent years 336:347 recent years 336:347 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 4 47 theme glycosyl 789:796 arg1 acceptors 798:806 various glycosyl acceptors 781:806 various glycosyl acceptors 781:806 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 48 contain carrying 719:726 arg2 groups 762:767 one to three unprotected hydroxyl groups 728:767 one to three unprotected hydroxyl groups 728:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 4 48 contain carrying 719:726 arg1 donors 712:717 glycosyl donors 703:717 glycosyl donors carrying one to three unprotected hydroxyl groups 703:767 Using dialkylboryl triflate as an in situ masking reagent, a wide array of glycosyl donors carrying one to three unprotected hydroxyl groups reacts with various glycosyl acceptors to furnish the desired products with good control over regioselectivity and stereoselectivity. 29079775 2 49 theme facile 413:418 arg1 assembly 420:427 facile assembly 413:427 facile assembly of glycan chains 413:444 Owing to the specific stereo-configuration at each anomeric linkage and diversity in branching, significant breakthroughs in recent years have focused on either stereoselective glycosylation methods or facile assembly of glycan chains. 29079775 1 50 theme complex 191:197 arg1 structures 199:208 these complex structures 185:208 these complex structures 185:208 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 29079775 0 51 with glycosylation 41:53 arg1 donors 72:77 unprotected donors 60:77 unprotected donors 60:77 A minimalist approach to stereoselective glycosylation with unprotected donors. 29079775 1 52 theme chemical 163:170 arg1 synthesis 172:180 the chemical synthesis 159:180 the chemical synthesis of these complex structures 159:208 Mechanistic study of carbohydrate interactions in biological systems calls for the chemical synthesis of these complex structures. 26457763 7 0 theme microarray 1207:1216 arg1 results 1218:1224 these microarray results 1201:1224 these microarray results 1201:1224 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 4 1 theme viral 653:657 arg1 control 659:665 viral control 653:665 viral control 653:665 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 1 2 theme PEMV 138:141 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 6 3 theme fluorous 998:1005 arg1 tags 1007:1010 fluorous tags 998:1010 fluorous tags 998:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 4 4 theme control 659:665 arg1 possibility 638:648 the possibility 634:648 the possibility of viral control 634:665 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 6 5 theme terminal 970:977 arg1 trimannosides 979:991 The resulting insect N-glycan terminal trimannosides 940:991 The resulting insect N-glycan terminal trimannosides with fluorous tags 940:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 6 6 with binding 1058:1064 arg1 PEMV 1106:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 0 7 theme Enation 90:96 arg1 Virus 105:109 Pea Enation Mosaic Virus 86:109 Pea Enation Mosaic Virus 86:109 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 8 8 theme glycan-PEMV 1428:1438 arg1 interaction 1440:1450 this particular glycan-PEMV interaction 1412:1450 this particular glycan-PEMV interaction 1412:1450 The ITC studies confirmed the microarray results and suggested that this particular glycan-PEMV interaction is not involved in virus uptake and transport through the aphid. 26457763 8 9 theme ITC 1348:1350 arg1 studies 1352:1358 The ITC studies 1344:1358 The ITC studies 1344:1358 The ITC studies confirmed the microarray results and suggested that this particular glycan-PEMV interaction is not involved in virus uptake and transport through the aphid. 26457763 4 10 theme terminal 737:744 arg1 trimannosides 746:758 insect N-glycan terminal trimannosides 721:758 insect N-glycan terminal trimannosides 721:758 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 3 11 from release 522:528 arg1 saliva 533:538 saliva 533:538 saliva 533:538 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 0 12 theme Pea 86:88 arg1 Virus 105:109 Pea Enation Mosaic Virus 86:109 Pea Enation Mosaic Virus 86:109 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 3 13 gly glycosylated 380:391 arg1 N 417:417 a heavily glycosylated receptor aminopeptidase N 370:417 a heavily glycosylated receptor aminopeptidase N 370:417 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 5 14 with features 809:816 arg1 ester 852:856 C-5 ester 848:856 C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension 848:937 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 4 15 theme N-glycan 728:735 arg1 trimannosides 746:758 insect N-glycan terminal trimannosides 721:758 insect N-glycan terminal trimannosides 721:758 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 4 16 theme glycan 686:691 arg1 interaction 693:703 a glycan interaction 684:703 a glycan interaction 684:703 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 0 17 theme Virus 105:109 arg1 Affinity 74:81 the Binding Affinity 62:81 the Binding Affinity of Pea Enation Mosaic Virus 62:109 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 5 18 theme building 828:835 arg1 block 837:841 a mannose building block 818:841 a mannose building block 818:841 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 4 19 theme glycans 587:593 arg1 role 579:582 the role 575:582 the role of glycans in PEMV-aphid interactions 575:620 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 2 20 theme aphid-virus 250:260 arg1 interactions 262:273 the aphid-virus interactions 246:273 the aphid-virus interactions required for disease transmission 246:307 However, the aphid-virus interactions required for disease transmission are poorly understood. 26457763 1 21 theme --a 143:145 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 0 22 theme Mosaic 98:103 arg1 Virus 105:109 Pea Enation Mosaic Virus 86:109 Pea Enation Mosaic Virus 86:109 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 3 23 theme aminopeptidase 402:415 arg1 N 417:417 a heavily glycosylated receptor aminopeptidase N 370:417 a heavily glycosylated receptor aminopeptidase N 370:417 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 1 24 dep Pea 112:114 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 1 25 theme plant 147:151 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 0 26 theme Solution-Phase 10:23 arg1 Synthesis 25:33 Automated Solution-Phase Synthesis 0:33 Automated Solution-Phase Synthesis of Insect Glycans 0:51 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 6 27 used used 1017:1020 arg2 trimannosides 979:991 The resulting insect N-glycan terminal trimannosides 940:991 The resulting insect N-glycan terminal trimannosides with fluorous tags 940:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 5 28 theme subsequent 912:921 arg1 extension 929:937 subsequent chain extension 912:937 subsequent chain extension 912:937 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 1 29 theme RNA 153:155 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 0 30 theme Automated 0:8 arg1 Synthesis 25:33 Automated Solution-Phase Synthesis 0:33 Automated Solution-Phase Synthesis of Insect Glycans 0:51 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 6 31 theme N-glycan 961:968 arg1 trimannosides 979:991 The resulting insect N-glycan terminal trimannosides 940:991 The resulting insect N-glycan terminal trimannosides with fluorous tags 940:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 5 32 theme C-5 848:850 arg1 ester 852:856 C-5 ester 848:856 C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension 848:937 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 5 33 theme chain 923:927 arg1 extension 929:937 subsequent chain extension 912:937 subsequent chain extension 912:937 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 7 34 theme isothermal 1282:1291 arg1 calorimetry 1303:1313 isothermal titration calorimetry 1282:1313 isothermal titration calorimetry studies with unlabeled PEMV 1282:1341 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 7 35 theme unlabeled 1328:1336 arg1 PEMV 1338:1341 unlabeled PEMV 1328:1341 unlabeled PEMV 1328:1341 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 6 36 theme insect 954:959 arg1 trimannosides 979:991 The resulting insect N-glycan terminal trimannosides 940:991 The resulting insect N-glycan terminal trimannosides with fluorous tags 940:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 8 37 theme virus 1471:1475 arg1 uptake 1477:1482 virus uptake 1471:1482 virus uptake 1471:1482 The ITC studies confirmed the microarray results and suggested that this particular glycan-PEMV interaction is not involved in virus uptake and transport through the aphid. 26457763 3 38 dep aphid 547:551 arg1 feeds 553:557 feeds 553:557 feeds 553:557 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 2 39 theme disease 288:294 arg1 transmission 296:307 disease transmission 288:307 disease transmission 288:307 However, the aphid-virus interactions required for disease transmission are poorly understood. 26457763 5 40 theme mannose 820:826 arg1 block 837:841 a mannose building block 818:841 a mannose building block 818:841 The route features a mannose building block with C-5 ester enforcing a β-linkage, which also provides a site for subsequent chain extension. 26457763 6 41 theme insect 1153:1158 arg1 glycan 1160:1165 the insect glycan 1149:1165 the insect glycan 1149:1165 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 6 42 theme resulting 944:952 arg1 trimannosides 979:991 The resulting insect N-glycan terminal trimannosides 940:991 The resulting insect N-glycan terminal trimannosides with fluorous tags 940:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 3 43 theme virus 336:340 arg1 transmission 342:353 virus transmission 336:353 virus transmission 336:353 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 6 44 theme isothiocyanate-labeled 1083:1104 arg1 PEMV 1106:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 3 45 theme aphid 495:499 arg1 cavity 506:511 the aphid body cavity 491:511 the aphid body cavity prior to release in saliva 491:538 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 0 46 theme Glycans 45:51 arg1 Synthesis 25:33 Automated Solution-Phase Synthesis 0:33 Automated Solution-Phase Synthesis of Insect Glycans 0:51 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 7 47 theme titration 1293:1301 arg1 calorimetry 1303:1313 isothermal titration calorimetry 1282:1313 isothermal titration calorimetry studies with unlabeled PEMV 1282:1341 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 7 48 theme fluorous 1242:1249 arg1 tag 1251:1253 the fluorous tag 1238:1253 the fluorous tag 1238:1253 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 6 49 theme fluorous 1027:1034 arg1 microarray 1036:1045 a fluorous microarray 1025:1045 a fluorous microarray 1025:1045 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 0 50 theme Insect 38:43 arg1 Glycans 45:51 Insect Glycans 38:51 Insect Glycans 38:51 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 7 51 with studies 1315:1321 arg1 PEMV 1338:1341 unlabeled PEMV 1328:1341 unlabeled PEMV 1328:1341 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 6 52 theme fluorescein 1071:1081 arg1 PEMV 1106:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 fluorescein isothiocyanate-labeled PEMV 1071:1109 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 1 53 theme aphids--causes 190:203 arg1 disease 205:211 aphids--causes disease 190:211 aphids--causes disease in multiple food crops 190:234 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 3 54 theme glycosylated 380:391 arg1 N 417:417 a heavily glycosylated receptor aminopeptidase N 370:417 a heavily glycosylated receptor aminopeptidase N 370:417 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 8 55 theme microarray 1374:1383 arg1 results 1385:1391 the microarray results 1370:1391 the microarray results 1370:1391 The ITC studies confirmed the microarray results and suggested that this particular glycan-PEMV interaction is not involved in virus uptake and transport through the aphid. 26457763 3 56 theme pea 426:428 arg1 gut 436:438 the pea aphid gut 422:438 the pea aphid gut 422:438 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 8 57 theme particular 1417:1426 arg1 interaction 1440:1450 this particular glycan-PEMV interaction 1412:1450 this particular glycan-PEMV interaction 1412:1450 The ITC studies confirmed the microarray results and suggested that this particular glycan-PEMV interaction is not involved in virus uptake and transport through the aphid. 26457763 3 58 theme gut 471:473 arg1 epithelium 475:484 the gut epithelium 467:484 the gut epithelium into the aphid body cavity prior to release in saliva 467:538 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 3 59 theme receptor 393:400 arg1 N 417:417 a heavily glycosylated receptor aminopeptidase N 370:417 a heavily glycosylated receptor aminopeptidase N 370:417 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 6 60 theme specific 1124:1131 arg1 binding 1133:1139 no specific binding 1121:1139 no specific binding between the insect glycan and PEMV 1121:1174 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 1 61 from disease 205:211 arg1 crops 230:234 multiple food crops 216:234 multiple food crops 216:234 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 6 62 with trimannosides 979:991 arg1 tags 1007:1010 fluorous tags 998:1010 fluorous tags 998:1010 The resulting insect N-glycan terminal trimannosides with fluorous tags were used in a fluorous microarray to analyze binding with fluorescein isothiocyanate-labeled PEMV; however, no specific binding between the insect glycan and PEMV was detected. 26457763 4 63 theme solution-phase 773:786 arg1 synthesis 788:796 automated solution-phase synthesis 763:796 automated solution-phase synthesis 763:796 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 1 64 theme enation 116:122 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 4 65 theme insect 721:726 arg1 trimannosides 746:758 insect N-glycan terminal trimannosides 721:758 insect N-glycan terminal trimannosides 721:758 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 3 66 theme prior 513:517 arg1 cavity 506:511 the aphid body cavity 491:511 the aphid body cavity prior to release in saliva 491:538 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 4 67 theme automated 763:771 arg1 synthesis 788:796 automated solution-phase synthesis 763:796 automated solution-phase synthesis 763:796 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 1 68 theme mosaic 124:129 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 0 69 theme Binding 66:72 arg1 Affinity 74:81 the Binding Affinity 62:81 the Binding Affinity of Pea Enation Mosaic Virus 62:109 Automated Solution-Phase Synthesis of Insect Glycans to Probe the Binding Affinity of Pea Enation Mosaic Virus. 26457763 4 70 theme PEMV-aphid 598:607 arg1 interactions 609:620 PEMV-aphid interactions 598:620 PEMV-aphid interactions 598:620 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 7 71 theme calorimetry 1303:1313 arg1 studies 1315:1321 isothermal titration calorimetry studies 1282:1321 isothermal titration calorimetry studies with unlabeled PEMV 1282:1341 To confirm these microarray results, we removed the fluorous tag from the trimannosides for isothermal titration calorimetry studies with unlabeled PEMV. 26457763 1 72 theme virus 131:135 arg1 virus 157:161 enation mosaic virus (PEMV)--a plant RNA virus 116:161 Pea enation mosaic virus (PEMV)--a plant RNA virus 112:161 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 3 73 theme aphid 430:434 arg1 gut 436:438 the pea aphid gut 422:438 the pea aphid gut 422:438 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 1 74 theme multiple 216:223 arg1 crops 230:234 multiple food crops 216:234 multiple food crops 216:234 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 26457763 4 75 from role 579:582 arg1 interactions 609:620 PEMV-aphid interactions 598:620 PEMV-aphid interactions 598:620 To investigate the role of glycans in PEMV-aphid interactions and explore the possibility of viral control through blocking a glycan interaction, we synthesized insect N-glycan terminal trimannosides by automated solution-phase synthesis. 26457763 3 76 theme body 501:504 arg1 cavity 506:511 the aphid body cavity 491:511 the aphid body cavity prior to release in saliva 491:538 For virus transmission, PEMV binds to a heavily glycosylated receptor aminopeptidase N in the pea aphid gut and is transcytosed across the gut epithelium into the aphid body cavity prior to release in saliva as the aphid feeds. 26457763 1 77 theme food 225:228 arg1 crops 230:234 multiple food crops 216:234 multiple food crops 216:234 Pea enation mosaic virus (PEMV)--a plant RNA virus transmitted exclusively by aphids--causes disease in multiple food crops. 24725217 12 0 theme NMR 1822:1824 arg1 importance 1770:1779 the importance 1766:1779 the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control 1766:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 7 1 from separation 1100:1109 arg1 overlapping 1067:1077 overlapping 1067:1077 overlapping 1067:1077 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 2 2 theme antibodies 358:367 arg1 glycosylation 329:341 nonhuman glycosylation 320:341 nonhuman glycosylation of recombinant antibodies 320:367 Recently, it was found that nonhuman glycosylation of recombinant antibodies can cause tremendous problems for some patients. 24725217 6 3 theme three-dimensional 948:964 arg1 3DCC 985:988 3DCC 985:988 3DCC 985:988 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 6 3 theme three-dimensional 948:964 arg1 correlation 972:982 three-dimensional cross correlation 948:982 three-dimensional cross correlation (3DCC) 948:989 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 4 4 theme N-glycan 618:625 arg1 analysis 627:634 detailed N-glycan analysis 609:634 detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS 609:684 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 7 5 theme chromatographic 1170:1184 arg1 fractions 1186:1194 chromatographic fractions 1170:1194 chromatographic fractions 1170:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 8 6 theme branching 1251:1259 arg1 position 1261:1268 the branching position 1247:1268 the branching position of one monosaccharide unit 1247:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 7 7 theme NMR 1144:1146 arg1 spectra 1148:1154 the NMR spectra 1140:1154 the NMR spectra obtained from chromatographic fractions 1140:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 4 8 theme cetuximab 664:672 arg1 analysis 627:634 detailed N-glycan analysis 609:634 detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS 609:684 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 8 9 theme monosaccharide 1277:1290 arg1 unit 1292:1295 one monosaccharide unit 1273:1295 one monosaccharide unit 1273:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 12 10 from development 1886:1896 arg1 analysis 1842:1849 the glycome analysis 1830:1849 the glycome analysis of biopharmaceuticals in research, development, and quality control 1830:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 11 from research 1876:1883 arg1 analysis 1842:1849 the glycome analysis 1830:1849 the glycome analysis of biopharmaceuticals in research, development, and quality control 1830:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 7 12 theme chromatographic 1084:1098 arg1 separation 1100:1109 a chromatographic separation 1082:1109 a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1082:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 7 13 theme N-glycans 1039:1047 arg1 spectra 1023:1029 extract NMR spectra 1011:1029 extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1011:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 1 14 theme Monoclonal 127:136 arg1 antibodies 138:147 Monoclonal antibodies 127:147 Monoclonal antibodies 127:147 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 12 15 from biopharmaceuticals 1854:1871 arg1 research 1876:1883 research 1876:1883 research 1876:1883 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 15 from biopharmaceuticals 1854:1871 arg1 control 1911:1917 quality control 1903:1917 quality control 1903:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 15 from biopharmaceuticals 1854:1871 arg1 development 1886:1896 development 1886:1896 development 1886:1896 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 4 16 theme unambiguous 738:748 arg1 structures 750:759 unambiguous structures 738:759 unambiguous structures 738:759 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 12 17 theme use 1795:1797 arg1 importance 1770:1779 the importance 1766:1779 the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control 1766:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 7 18 theme NMR 1019:1021 arg1 spectra 1023:1029 extract NMR spectra 1011:1029 extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1011:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 5 19 theme Thirty-seven 762:773 arg1 compositions 784:795 Thirty-seven N-glycan compositions 762:795 Thirty-seven N-glycan compositions 762:795 Thirty-seven N-glycan compositions were identified by LC-MS(/MS). 24725217 4 20 theme H 722:722 arg1 NMR 724:726 (1)H NMR 719:726 (1)H NMR 719:726 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 4 21 from analysis 627:634 arg1 results 584:590 results 584:590 results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures 584:759 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 9 22 theme method 1380:1385 arg1 improvement 1356:1366 an improvement 1353:1366 an improvement of the 3DCC method 1353:1385 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 12 23 from control 1911:1917 arg1 analysis 1842:1849 the glycome analysis 1830:1849 the glycome analysis of biopharmaceuticals in research, development, and quality control 1830:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 24 dep NMR 1822:1824 arg1 1 1818:1818 1 1818:1818 1 1818:1818 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 5 25 theme N-glycan 775:782 arg1 compositions 784:795 Thirty-seven N-glycan compositions 762:795 Thirty-seven N-glycan compositions 762:795 Thirty-seven N-glycan compositions were identified by LC-MS(/MS). 24725217 0 26 theme Unambiguous 0:10 arg1 characterization 12:27 Unambiguous characterization 0:27 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 0:125 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 9 27 theme value 1411:1415 arg1 SVD 1432:1434 SVD 1432:1434 SVD 1432:1434 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 9 27 theme value 1411:1415 arg1 decomposition 1417:1429 singular value decomposition 1402:1429 singular value decomposition (SVD) 1402:1435 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 3 28 theme therapeutic 484:494 arg1 antibodies 496:505 therapeutic antibodies 484:505 therapeutic antibodies 484:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 4 29 dep NMR 724:726 arg1 1 720:720 1 720:720 1 720:720 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 3 30 theme pattern 473:479 arg1 assignment 441:450 unambiguous assignment 429:450 unambiguous assignment of the glycosylation pattern of therapeutic antibodies 429:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 12 31 theme glycome 1834:1840 arg1 analysis 1842:1849 the glycome analysis 1830:1849 the glycome analysis of biopharmaceuticals in research, development, and quality control 1830:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 3 32 theme high 513:516 arg1 importance 518:527 high importance 513:527 high importance 513:527 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 0 33 theme antibody 56:63 arg1 cetuximab 65:73 monoclonal antibody cetuximab 45:73 monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy 45:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 12 34 theme biopharmaceuticals 1854:1871 arg1 analysis 1842:1849 the glycome analysis 1830:1849 the glycome analysis of biopharmaceuticals in research, development, and quality control 1830:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 35 from analysis 1842:1849 arg1 research 1876:1883 research 1876:1883 research 1876:1883 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 35 from analysis 1842:1849 arg1 control 1911:1917 quality control 1903:1917 quality control 1903:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 35 from analysis 1842:1849 arg1 development 1886:1896 development 1886:1896 development 1886:1896 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 3 36 gly glycosylation 459:471 arg1 antibodies 496:505 therapeutic antibodies 484:505 therapeutic antibodies 484:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 12 37 theme H 1820:1820 arg1 NMR 1822:1824 (1)H NMR 1817:1824 (1)H NMR 1817:1824 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 2 38 theme tremendous 379:388 arg1 problems 390:397 tremendous problems 379:397 tremendous problems for some patients 379:415 Recently, it was found that nonhuman glycosylation of recombinant antibodies can cause tremendous problems for some patients. 24725217 1 39 theme other 239:243 arg1 inflammation 278:289 inflammation 278:289 inflammation 278:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 1 39 theme other 239:243 arg1 diseases 245:252 various other diseases 231:252 various other diseases such as autoimmunity or inflammation 231:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 1 39 theme other 239:243 arg1 autoimmunity 262:273 autoimmunity 262:273 autoimmunity 262:273 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 8 40 theme Even 1197:1200 arg1 structures 1216:1225 Even mass isobaric structures 1197:1225 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit 1197:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 11 41 theme glycans 1656:1662 arg1 glycans 1656:1662 all detected glycans 1643:1662 all detected glycans 1643:1662 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 11 41 theme glycans 1656:1662 arg1 % 1638:1638 24% 1636:1638 24% of all detected glycans 1636:1662 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 8 42 theme isobaric 1207:1214 arg1 structures 1216:1225 Even mass isobaric structures 1197:1225 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit 1197:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 6 43 theme cross 966:970 arg1 3DCC 985:988 3DCC 985:988 3DCC 985:988 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 6 43 theme cross 966:970 arg1 correlation 972:982 three-dimensional cross correlation 948:982 three-dimensional cross correlation (3DCC) 948:989 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 11 44 theme immunogenic 1677:1687 arg1 epitope 1699:1705 the immunogenic α-1,3-Gal epitope 1673:1705 the immunogenic α-1,3-Gal epitope 1673:1705 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 4 45 theme detailed 609:616 arg1 analysis 627:634 detailed N-glycan analysis 609:634 detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS 609:684 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 2 46 theme recombinant 346:356 arg1 antibodies 358:367 recombinant antibodies 346:367 recombinant antibodies 346:367 Recently, it was found that nonhuman glycosylation of recombinant antibodies can cause tremendous problems for some patients. 24725217 11 47 theme identified 1588:1597 arg1 glycans 1599:1605 the ten unambiguously identified glycans 1566:1605 the ten unambiguously identified glycans 1566:1605 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 3 48 theme compatibility 553:565 arg1 assessment 533:542 assessment 533:542 assessment of human compatibility 533:565 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 1 49 theme therapeutic 178:188 arg1 drugs 190:194 therapeutic drugs 178:194 therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation 178:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 6 50 theme introduced 923:932 arg1 method 934:939 the recently introduced method 910:939 the recently introduced method called three-dimensional cross correlation (3DCC) 910:989 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 12 51 theme LC-MS 1802:1806 arg1 NMR 1822:1824 (1)H NMR 1817:1824 (1)H NMR 1817:1824 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 12 51 theme LC-MS 1802:1806 arg1 use 1795:1797 integrated use 1784:1797 integrated use of LC-MS(/MS) 1784:1811 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 11 52 theme N-glycolylneuraminic 1714:1733 arg1 acid 1735:1738 N-glycolylneuraminic acid 1714:1738 N-glycolylneuraminic acid 1714:1738 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 10 53 theme smallest 1469:1476 arg1 amount 1478:1483 The smallest amount 1465:1483 The smallest amount of the N-glycan characterized by 3DCC 1465:1521 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 10 53 theme smallest 1469:1476 arg1 N-glycan 1492:1499 the N-glycan 1488:1499 the N-glycan characterized by 3DCC 1488:1521 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 10 53 theme smallest 1469:1476 arg1 pmol 1545:1548 approximately 400 pmol 1527:1548 approximately 400 pmol (836 ng) 1527:1557 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 11 54 contain possess 1665:1671 arg1 N-glycans 1612:1620 six N-glycans 1608:1620 six N-glycans 1608:1620 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 11 54 contain possess 1665:1671 arg2 acid 1735:1738 N-glycolylneuraminic acid 1714:1738 N-glycolylneuraminic acid 1714:1738 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 11 54 contain possess 1665:1671 arg2 epitope 1699:1705 the immunogenic α-1,3-Gal epitope 1673:1705 the immunogenic α-1,3-Gal epitope 1673:1705 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 4 55 theme antibody 655:662 arg1 cetuximab 664:672 the therapeutic antibody cetuximab 639:672 the therapeutic antibody cetuximab 639:672 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 1 56 theme cancer 217:222 arg1 treatment 204:212 the treatment 200:212 the treatment of cancer 200:222 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 4 57 dep broad 599:603 arg1 analyses 686:693 analyses 686:693 analyses 686:693 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 8 58 theme unit 1292:1295 arg1 position 1261:1268 the branching position 1247:1268 the branching position of one monosaccharide unit 1247:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 7 59 theme pure 1034:1037 arg1 N-glycans 1039:1047 pure N-glycans 1034:1047 pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1034:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 12 60 theme integrated 1784:1793 arg1 use 1795:1797 integrated use 1784:1797 integrated use of LC-MS(/MS) 1784:1811 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 0 61 theme LC-MS/MS 93:100 arg1 integration 78:88 integration 78:88 integration of LC-MS/MS and ¹H NMR spectroscopy 78:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 4 62 from broad 599:603 arg1 results 584:590 results 584:590 results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures 584:759 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 0 63 theme ¹H 106:107 arg1 spectroscopy 113:124 ¹H NMR spectroscopy 106:124 ¹H NMR spectroscopy 106:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 9 64 theme 3DCC 1375:1378 arg1 method 1380:1385 the 3DCC method 1371:1385 the 3DCC method 1371:1385 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 7 65 theme extract 1011:1017 arg1 spectra 1023:1029 extract NMR spectra 1011:1029 extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1011:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 10 66 theme N-glycan 1492:1499 arg1 amount 1478:1483 The smallest amount 1465:1483 The smallest amount of the N-glycan characterized by 3DCC 1465:1521 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 10 66 theme N-glycan 1492:1499 arg1 N-glycan 1492:1499 the N-glycan 1488:1499 the N-glycan characterized by 3DCC 1488:1521 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 10 66 theme N-glycan 1492:1499 arg1 pmol 1545:1548 approximately 400 pmol 1527:1548 approximately 400 pmol (836 ng) 1527:1557 The smallest amount of the N-glycan characterized by 3DCC was approximately 400 pmol (836 ng). 24725217 0 67 theme spectroscopy 113:124 arg1 integration 78:88 integration 78:88 integration of LC-MS/MS and ¹H NMR spectroscopy 78:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 1 68 theme various 231:237 arg1 inflammation 278:289 inflammation 278:289 inflammation 278:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 1 68 theme various 231:237 arg1 diseases 245:252 various other diseases 231:252 various other diseases such as autoimmunity or inflammation 231:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 1 68 theme various 231:237 arg1 autoimmunity 262:273 autoimmunity 262:273 autoimmunity 262:273 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 24725217 3 69 theme glycosylation 459:471 arg1 pattern 473:479 the glycosylation pattern 455:479 the glycosylation pattern of therapeutic antibodies 455:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 9 70 theme singular 1402:1409 arg1 SVD 1432:1434 SVD 1432:1434 SVD 1432:1434 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 9 70 theme singular 1402:1409 arg1 decomposition 1417:1429 singular value decomposition 1402:1429 singular value decomposition (SVD) 1402:1435 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 0 71 theme N-glycans 32:40 arg1 characterization 12:27 Unambiguous characterization 0:27 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 0:125 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 3 72 theme antibodies 496:505 arg1 pattern 473:479 the glycosylation pattern 455:479 the glycosylation pattern of therapeutic antibodies 455:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 0 73 theme monoclonal 45:54 arg1 cetuximab 65:73 monoclonal antibody cetuximab 45:73 monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy 45:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 12 74 theme quality 1903:1909 arg1 control 1911:1917 quality control 1903:1917 quality control 1903:1917 These results illustrate the importance of integrated use of LC-MS(/MS) and (1)H NMR for the glycome analysis of biopharmaceuticals in research, development, and quality control. 24725217 3 75 theme unambiguous 429:439 arg1 assignment 441:450 unambiguous assignment 429:450 unambiguous assignment of the glycosylation pattern of therapeutic antibodies 429:505 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 0 76 theme NMR 109:111 arg1 spectroscopy 113:124 ¹H NMR spectroscopy 106:124 ¹H NMR spectroscopy 106:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 0 77 theme cetuximab 65:73 arg1 N-glycans 32:40 N-glycans 32:40 N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy 32:124 Unambiguous characterization of N-glycans of monoclonal antibody cetuximab by integration of LC-MS/MS and ¹H NMR spectroscopy. 24725217 7 78 from overlapping 1067:1077 arg1 separation 1100:1109 a chromatographic separation 1082:1109 a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions 1082:1194 It was possible to extract NMR spectra of pure N-glycans that were heavily overlapping in a chromatographic separation by mathematically dissecting the NMR spectra obtained from chromatographic fractions. 24725217 2 79 gly glycosylation 329:341 arg1 antibodies 358:367 recombinant antibodies 346:367 recombinant antibodies 346:367 Recently, it was found that nonhuman glycosylation of recombinant antibodies can cause tremendous problems for some patients. 24725217 2 80 theme nonhuman 320:327 arg1 glycosylation 329:341 nonhuman glycosylation 320:341 nonhuman glycosylation of recombinant antibodies 320:367 Recently, it was found that nonhuman glycosylation of recombinant antibodies can cause tremendous problems for some patients. 24725217 9 81 theme data 1459:1462 arg1 processing 1441:1450 processing 1441:1450 processing of the data 1441:1462 We also developed an improvement of the 3DCC method by introducing singular value decomposition (SVD) for processing of the data. 24725217 8 82 theme mass 1202:1205 arg1 structures 1216:1225 Even mass isobaric structures 1197:1225 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit 1197:1295 Even mass isobaric structures that differ only in the branching position of one monosaccharide unit were distinguished and characterized. 24725217 6 83 theme abundant 846:853 arg1 structures 855:864 ten abundant structures 842:864 ten abundant structures 842:864 Subsequently, ten abundant structures were structurally characterized by applying the recently introduced method called three-dimensional cross correlation (3DCC). 24725217 4 84 theme therapeutic 643:653 arg1 cetuximab 664:672 the therapeutic antibody cetuximab 639:672 the therapeutic antibody cetuximab 639:672 Here we present results from a broad and detailed N-glycan analysis of the therapeutic antibody cetuximab by LC-MS/MS analyses tightly integrated with (1)H NMR to obtain unambiguous structures. 24725217 11 85 theme α-1,3-Gal 1689:1697 arg1 epitope 1699:1705 the immunogenic α-1,3-Gal epitope 1673:1705 the immunogenic α-1,3-Gal epitope 1673:1705 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 11 86 theme detected 1647:1654 arg1 glycans 1656:1662 all detected glycans 1643:1662 all detected glycans 1643:1662 Among the ten unambiguously identified glycans, six N-glycans, representing 24% of all detected glycans, possess the immunogenic α-1,3-Gal epitope and/or N-glycolylneuraminic acid. 24725217 3 87 theme human 547:551 arg1 compatibility 553:565 human compatibility 547:565 human compatibility 547:565 Therefore, unambiguous assignment of the glycosylation pattern of therapeutic antibodies is of high importance for assessment of human compatibility. 24725217 1 88 theme diseases 245:252 arg1 drugs 190:194 therapeutic drugs 178:194 therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation 178:289 Monoclonal antibodies are most rapidly emerging as therapeutic drugs for the treatment of cancer and of various other diseases such as autoimmunity or inflammation. 26082231 5 0 theme distinct 750:757 arg1 one 828:830 one 828:830 one 828:830 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 0 theme distinct 750:757 arg1 other 893:897 other 893:897 other 893:897 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 0 theme distinct 750:757 arg1 ways 759:762 detail two distinct ways 739:762 detail two distinct ways in which to usefully interfere with oligosaccharide processing 739:825 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 4 1 gly glycoproteins 603:615 arg1 glycoproteins 603:615 glycoproteins 603:615 glycoproteins 603:615 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 6 2 gly glycoproteins 1228:1240 arg1 glycoproteins 1228:1240 glycoproteins 1228:1240 glycoproteins 1228:1240 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 6 3 dep controlled 1052:1061 arg1 radical 1064:1070 radical 1064:1070 radical 1064:1070 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 5 4 theme gene 938:941 arg1 loci 943:946 gene loci 938:946 gene loci that control oligosaccharide processing 938:986 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 5 theme cell 916:919 arg1 lines 921:925 cell lines 916:925 cell lines mutated at gene loci that control oligosaccharide processing 916:986 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 4 6 theme present 592:598 arg1 N-glycans 582:590 the N-glycans 578:590 the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal 578:715 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 6 7 contain have 1030:1033 arg1 approaches 1019:1028 Both approaches 1014:1028 Both approaches 1014:1028 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 6 7 contain have 1030:1033 arg2 capacity 1039:1046 the capacity 1035:1046 the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression 1035:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 6 8 theme controlled 1052:1061 arg1 alteration 1072:1081 controlled, radical alteration 1052:1081 controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression 1052:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 3 9 theme other 367:371 arg1 hand 373:376 the other hand 363:376 the other hand 363:376 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 3 10 theme large 395:399 arg1 carbohydrate 412:423 carbohydrate 412:423 carbohydrate 412:423 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 3 10 theme large 395:399 arg1 amounts 401:407 large amounts 395:407 large amounts of carbohydrate 395:423 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 3 11 theme structural 446:455 arg1 analysis 472:479 the structural and functional analysis 442:479 the structural and functional analysis of glycoproteins 442:496 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 2 12 theme glycoforms 335:344 arg1 biosynthesis 308:319 the biosynthesis 304:319 the biosynthesis of particular glycoforms 304:344 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 5 13 theme specific 853:860 arg1 inhibitors 873:882 specific processing inhibitors 853:882 specific processing inhibitors 853:882 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 0 14 theme Glycan 0:5 arg1 Remodeling 7:16 Glycan Remodeling 0:16 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells 0:81 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 2 15 theme particular 324:333 arg1 glycoforms 335:344 particular glycoforms 324:344 particular glycoforms 324:344 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 5 16 theme processing 862:871 arg1 inhibitors 873:882 specific processing inhibitors 853:882 specific processing inhibitors 853:882 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 0 17 theme Processing 23:32 arg1 Inhibitors 34:43 Processing Inhibitors 23:43 Processing Inhibitors 23:43 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 1 18 from molecules 127:135 arg1 biology 149:155 metazoan biology 140:155 metazoan biology 140:155 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 1 19 theme important 101:109 arg1 molecules 127:135 the most important and interesting molecules 92:135 the most important and interesting molecules in metazoan biology 92:155 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 5 20 theme lines 921:925 arg1 selection 903:911 the selection 899:911 the selection of cell lines mutated at gene loci that control oligosaccharide processing 899:986 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 6 21 theme great 1184:1188 arg1 utility 1190:1196 great utility 1184:1196 great utility 1184:1196 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 3 22 gly glycoproteins 484:496 arg1 glycoproteins 484:496 glycoproteins 484:496 glycoproteins 484:496 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 6 23 theme protein 1155:1161 arg1 expression 1163:1172 heterologous protein expression 1142:1172 heterologous protein expression 1142:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 3 24 theme carbohydrate 412:423 arg1 carbohydrate 412:423 carbohydrate 412:423 carbohydrate 412:423 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 3 24 theme carbohydrate 412:423 arg1 amounts 401:407 large amounts 395:407 large amounts of carbohydrate 395:423 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 6 25 theme processing 1102:1111 arg1 alteration 1072:1081 controlled, radical alteration 1052:1081 controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression 1052:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 5 26 theme cytotoxic 995:1003 arg1 lectins 1005:1011 cytotoxic lectins 995:1011 cytotoxic lectins 995:1011 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 27 theme oligosaccharide 800:814 arg1 processing 816:825 oligosaccharide processing 800:825 oligosaccharide processing 800:825 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 3 28 theme functional 461:470 arg1 analysis 472:479 the structural and functional analysis 442:479 the structural and functional analysis of glycoproteins 442:496 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 0 29 theme Eukaryotic 66:75 arg1 Cells 77:81 Lectin-Resistant Eukaryotic Cells 49:81 Lectin-Resistant Eukaryotic Cells 49:81 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 6 30 theme oligosaccharide 1086:1100 arg1 processing 1102:1111 oligosaccharide processing 1086:1111 oligosaccharide processing 1086:1111 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 1 31 theme interesting 115:125 arg1 molecules 127:135 the most important and interesting molecules 92:135 the most important and interesting molecules in metazoan biology 92:155 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 0 32 theme Lectin-Resistant 49:64 arg1 Cells 77:81 Lectin-Resistant Eukaryotic Cells 49:81 Lectin-Resistant Eukaryotic Cells 49:81 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 2 33 theme structures 230:239 arg1 component 211:219 the carbohydrate component 194:219 the carbohydrate component of these structures 194:239 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 0 34 with Remodeling 7:16 arg1 Cells 77:81 Lectin-Resistant Eukaryotic Cells 49:81 Lectin-Resistant Eukaryotic Cells 49:81 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 0 34 with Remodeling 7:16 arg1 Inhibitors 34:43 Processing Inhibitors 23:43 Processing Inhibitors 23:43 Glycan Remodeling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells. 26082231 4 35 theme subsequent 698:707 arg1 removal 709:715 their subsequent removal 692:715 their subsequent removal 692:715 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 6 36 theme structural 1205:1214 arg1 analysis 1216:1223 the structural analysis 1201:1223 the structural analysis of glycoproteins 1201:1240 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 5 37 theme oligosaccharide 961:975 arg1 processing 977:986 oligosaccharide processing 961:986 oligosaccharide processing 961:986 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 6 38 theme glycoproteins 1228:1240 arg1 analysis 1216:1223 the structural analysis 1201:1223 the structural analysis of glycoproteins 1201:1240 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 3 39 theme glycoproteins 484:496 arg1 analysis 472:479 the structural and functional analysis 442:479 the structural and functional analysis of glycoproteins 442:496 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 6 40 theme heterologous 1142:1153 arg1 expression 1163:1172 heterologous protein expression 1142:1172 heterologous protein expression 1142:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 1 41 gly glycoproteins 161:173 arg1 Some 84:87 Some 84:87 Some 84:87 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 1 41 gly glycoproteins 161:173 arg1 glycoproteins 161:173 glycoproteins 161:173 glycoproteins 161:173 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 1 41 gly glycoproteins 161:173 arg1 molecules 127:135 the most important and interesting molecules 92:135 the most important and interesting molecules in metazoan biology 92:155 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 2 42 theme component 211:219 arg1 importance 180:189 The importance 176:189 The importance of the carbohydrate component of these structures 176:239 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 2 43 theme disease 266:272 arg1 phenotypes 274:283 the disease phenotypes 262:283 the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted 262:357 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 1 44 theme metazoan 140:147 arg1 biology 149:155 metazoan biology 140:155 metazoan biology 140:155 Some of the most important and interesting molecules in metazoan biology are glycoproteins. 26082231 6 45 theme eukaryotic 1116:1125 arg1 cells 1127:1131 eukaryotic cells 1116:1131 eukaryotic cells used for heterologous protein expression 1116:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 5 46 theme detail 739:744 arg1 one 828:830 one 828:830 one 828:830 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 46 theme detail 739:744 arg1 other 893:897 other 893:897 other 893:897 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 5 46 theme detail 739:744 arg1 ways 759:762 detail two distinct ways 739:762 detail two distinct ways in which to usefully interfere with oligosaccharide processing 739:825 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 2 47 theme carbohydrate 198:209 arg1 component 211:219 the carbohydrate component 194:219 the carbohydrate component of these structures 194:239 The importance of the carbohydrate component of these structures is often revealed by the disease phenotypes that manifest when the biosynthesis of particular glycoforms is disrupted. 26082231 4 48 attach present 592:598 arg1 glycoproteins 603:615 glycoproteins 603:615 glycoproteins 603:615 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 4 48 attach present 592:598 arg2 N-glycans 582:590 the N-glycans 578:590 the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal 578:715 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 6 49 from alteration 1072:1081 arg1 cells 1127:1131 eukaryotic cells 1116:1131 eukaryotic cells used for heterologous protein expression 1116:1172 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 5 50 theme inhibitors 873:882 arg1 use 846:848 the use 842:848 the use of specific processing inhibitors 842:882 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 6 51 contain have 1179:1182 arg2 utility 1190:1196 great utility 1184:1196 great utility 1184:1196 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 6 51 contain have 1179:1182 arg1 approaches 1019:1028 Both approaches 1014:1028 Both approaches 1014:1028 Both approaches have the capacity for controlled, radical alteration of oligosaccharide processing in eukaryotic cells used for heterologous protein expression, and have great utility in the structural analysis of glycoproteins. 26082231 3 52 theme amounts 401:407 arg1 presence 383:390 the presence 379:390 the presence of large amounts of carbohydrate 379:423 On the other hand, the presence of large amounts of carbohydrate can often hinder the structural and functional analysis of glycoproteins. 26082231 4 53 theme good 515:518 arg1 reasons 520:526 good reasons 515:526 good reasons 515:526 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 26082231 5 54 dep one 828:830 arg1 selection 903:911 the selection 899:911 the selection of cell lines mutated at gene loci that control oligosaccharide processing 899:986 Here, we describe in detail two distinct ways in which to usefully interfere with oligosaccharide processing, one involving the use of specific processing inhibitors, and the other the selection of cell lines mutated at gene loci that control oligosaccharide processing, using cytotoxic lectins. 26082231 4 55 theme glycans 670:676 arg1 functions 653:661 the functions 649:661 the functions of the glycans 649:676 There are often good reasons, therefore, for wanting to engineer and predefine the N-glycans present on glycoproteins, e.g., in order to characterize the functions of the glycans or facilitate their subsequent removal. 24903886 7 0 theme many 1233:1236 arg1 strategies 1238:1247 many strategies 1233:1247 many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell 1233:1325 There are now many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell. 24903886 10 1 theme key 1549:1551 arg1 lectins 1600:1606 plant lectins 1594:1606 plant lectins that bind mammalian glycans with varying avidities 1594:1657 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 10 1 theme key 1549:1551 arg1 reagents 1553:1560 The key reagents 1545:1560 The key reagents for the protocols described 1545:1588 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 13 2 theme glycosylation 2139:2151 arg1 mutants 2153:2159 glycosylation mutants 2139:2159 glycosylation mutants 2139:2159 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 1 3 theme Glycosylation 108:120 arg1 engineering 122:132 Glycosylation engineering 108:132 Glycosylation engineering 108:132 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 8 4 from mutants 1399:1405 arg1 pathways 1424:1431 glycosylation pathways 1410:1431 glycosylation pathways 1410:1431 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 6 5 theme general 1152:1158 arg1 ways 1160:1163 more general ways 1147:1163 more general ways that usually affect several types of glycoconjugates 1147:1216 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 2 6 theme cell 292:295 arg1 mutant 311:316 a mammalian cell glycosylation mutant 280:316 a mammalian cell glycosylation mutant lacking a specific glycosyltransferase 280:355 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 11 7 with Cells 1714:1718 arg1 glycosylation 1733:1745 altered glycosylation 1725:1745 altered glycosylation 1725:1745 Cells with altered glycosylation generally become resistant or hypersensitive to lectin toxicity, and have reduced or increased lectin or antibody binding. 24903886 10 8 theme varying 1641:1647 arg1 avidities 1649:1657 varying avidities 1641:1657 varying avidities 1641:1657 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 11 9 theme antibody 1852:1859 arg1 binding 1861:1867 antibody binding 1852:1867 antibody binding 1852:1867 Cells with altered glycosylation generally become resistant or hypersensitive to lectin toxicity, and have reduced or increased lectin or antibody binding. 24903886 1 10 theme defined 211:217 arg1 complement 219:228 a more defined complement 204:228 a more defined complement of glycans on their glycoconjugates 204:264 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 5 11 dep glycans 979:985 arg1 elaborate 960:968 elaborate 960:968 elaborate 960:968 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 13 12 theme glycan 2291:2296 arg1 synthesis 2298:2306 glycan synthesis 2291:2306 glycan synthesis 2291:2306 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 6 13 theme glycoconjugates 1202:1216 arg1 types 1193:1197 several types 1185:1197 several types of glycoconjugates 1185:1216 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 11 14 gly glycosylation 1733:1745 arg1 Cells 1714:1718 Cells 1714:1718 Cells with altered glycosylation 1714:1745 Cells with altered glycosylation generally become resistant or hypersensitive to lectin toxicity, and have reduced or increased lectin or antibody binding. 24903886 1 15 gly glycoproteins 154:166 arg1 glycoproteins 154:166 glycoproteins 154:166 glycoproteins 154:166 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 9 16 theme altered 1513:1519 arg1 activity 1535:1542 an altered glycosylation activity 1510:1542 an altered glycosylation activity 1510:1542 This article will focus on how to rapidly characterize mammalian cells with an altered glycosylation activity. 24903886 6 17 theme other 1020:1024 arg1 transporters 1097:1108 transporters 1097:1108 transporters 1097:1108 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 6 17 theme other 1020:1024 arg1 synthases 1084:1092 nucleotide sugar synthases 1067:1092 nucleotide sugar synthases 1067:1092 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 6 17 theme other 1020:1024 arg1 genes 1026:1030 other genes 1020:1030 other genes 1020:1030 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 13 18 theme specific 2265:2272 arg1 gene 2274:2277 a specific gene 2263:2277 a specific gene involved in glycan synthesis 2263:2306 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 12 19 theme antibodies 1998:2007 arg1 assays 1893:1898 rapid assays 1887:1898 rapid assays to compare the cytotoxicity of lectins in a lectin resistance test 1887:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 12 19 theme antibodies 1998:2007 arg1 binding 1976:1982 the binding 1972:1982 the binding of lectins or antibodies by flow cytometry 1972:2025 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 5 20 contain have 908:911 arg1 glycoconjugates 887:901 glycoconjugates 887:901 glycoconjugates 887:901 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 5 20 contain have 908:911 arg2 glycans 913:919 glycans 913:919 glycans 913:919 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 0 21 theme altered 66:72 arg1 glycosylation 74:86 altered glycosylation 66:86 altered glycosylation in mammalian cells 66:105 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 13 22 theme glycosylation 2076:2088 arg1 changes 2090:2096 glycosylation changes 2076:2096 glycosylation changes expressed by a cell 2076:2116 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 5 23 theme additional 926:935 arg1 s 943:943 additional sugar(s) 926:944 additional sugar(s) that are more elaborate than the glycans of wild type cells 926:1004 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 4 24 theme gain-of-function 783:798 arg1 mutations 800:808 gain-of-function mutations 783:808 gain-of-function mutations that, for example, activate a silent glycosyltransferase gene 783:870 Another type of glycosylation mutant arises from gain-of-function mutations that, for example, activate a silent glycosyltransferase gene. 24903886 11 25 theme lectin 1795:1800 arg1 toxicity 1802:1809 lectin toxicity 1795:1809 lectin toxicity 1795:1809 Cells with altered glycosylation generally become resistant or hypersensitive to lectin toxicity, and have reduced or increased lectin or antibody binding. 24903886 12 26 theme lectin 1944:1949 arg1 test 1962:1965 a lectin resistance test 1942:1965 a lectin resistance test 1942:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 8 27 theme mammalian 1353:1361 arg1 cells 1363:1367 mammalian cells 1353:1367 mammalian cells 1353:1367 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 0 28 gly glycosylation 74:86 arg1 cells 101:105 mammalian cells 91:105 mammalian cells 91:105 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 6 29 theme nucleotide 1067:1076 arg1 synthases 1084:1092 nucleotide sugar synthases 1067:1092 nucleotide sugar synthases 1067:1092 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 9 30 with cells 1499:1503 arg1 activity 1535:1542 an altered glycosylation activity 1510:1542 an altered glycosylation activity 1510:1542 This article will focus on how to rapidly characterize mammalian cells with an altered glycosylation activity. 24903886 4 31 theme glycosylation 750:762 arg1 mutant 764:769 glycosylation mutant 750:769 glycosylation mutant 750:769 Another type of glycosylation mutant arises from gain-of-function mutations that, for example, activate a silent glycosyltransferase gene. 24903886 0 32 theme Rapid 0:4 arg1 assays 6:11 Rapid assays 0:11 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 0:106 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 13 33 theme or 2231:2232 arg1 mutation 2251:2258 a loss-of-function or gain-of-function mutation 2212:2258 a loss-of-function or gain-of-function mutation 2212:2258 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 7 34 theme mammalian 1312:1320 arg1 cell 1322:1325 a mammalian cell 1310:1325 a mammalian cell 1310:1325 There are now many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell. 24903886 6 35 from complement 1133:1142 arg1 ways 1160:1163 more general ways 1147:1163 more general ways that usually affect several types of glycoconjugates 1147:1216 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 12 36 theme rapid 1887:1891 arg1 assays 1893:1898 rapid assays 1887:1898 rapid assays to compare the cytotoxicity of lectins in a lectin resistance test 1887:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 2 37 gly glycoproteins 367:379 arg1 glycoproteins 367:379 glycoproteins 367:379 glycoproteins 367:379 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 7 38 theme glycosylation 1288:1300 arg1 gene 1302:1305 a glycosylation gene 1286:1305 a glycosylation gene 1286:1305 There are now many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell. 24903886 8 39 theme glycosylation 1410:1422 arg1 pathways 1424:1431 glycosylation pathways 1410:1431 glycosylation pathways 1410:1431 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 0 40 theme binding 37:43 arg1 changes 45:51 binding changes 37:51 binding changes 37:51 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 3 41 theme glycan 700:705 arg1 subset 707:712 a new or different glycan subset 681:712 a new or different glycan subset 681:712 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 2 42 theme specific 328:335 arg1 glycosyltransferase 337:355 a specific glycosyltransferase 326:355 a specific glycosyltransferase 326:355 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 5 43 theme type 995:998 arg1 cells 1000:1004 wild type cells 990:1004 wild type cells 990:1004 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 7 44 theme precise 1266:1272 arg1 mutation 1274:1281 a precise mutation 1264:1281 a precise mutation in a glycosylation gene 1264:1305 There are now many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell. 24903886 2 45 theme truncated 428:436 arg1 glycans 438:444 truncated glycans 428:444 truncated glycans missing the sugar transferred by that glycosyltransferase 428:502 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 1 46 theme glycans 233:239 arg1 complement 219:228 a more defined complement 204:228 a more defined complement of glycans on their glycoconjugates 204:264 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 3 47 theme mutant 721:726 arg1 cell 728:731 the mutant cell 717:731 the mutant cell 717:731 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 12 48 theme glycan-binding 2032:2045 arg1 assay 2047:2051 a glycan-binding assay 2030:2051 a glycan-binding assay 2030:2051 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 10 49 theme plant 1594:1598 arg1 lectins 1600:1606 plant lectins 1594:1606 plant lectins that bind mammalian glycans with varying avidities 1594:1657 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 10 49 theme plant 1594:1598 arg1 reagents 1553:1560 The key reagents 1545:1560 The key reagents for the protocols described 1545:1588 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 4 50 theme glycosyltransferase 847:865 arg1 gene 867:870 a silent glycosyltransferase gene 838:870 a silent glycosyltransferase gene 838:870 Another type of glycosylation mutant arises from gain-of-function mutations that, for example, activate a silent glycosyltransferase gene. 24903886 2 51 theme mammalian 282:290 arg1 mutant 311:316 a mammalian cell glycosylation mutant 280:316 a mammalian cell glycosylation mutant lacking a specific glycosyltransferase 280:355 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 1 52 used used 137:140 arg2 engineering 122:132 Glycosylation engineering 108:132 Glycosylation engineering 108:132 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 12 53 dep assays 1893:1898 arg1 compare 1903:1909 compare 1903:1909 to compare the cytotoxicity of lectins in a lectin resistance test 1900:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 12 54 theme flow 2012:2015 arg1 cytometry 2017:2025 flow cytometry 2012:2025 flow cytometry 2012:2025 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 10 55 theme mammalian 1618:1626 arg1 glycans 1628:1634 mammalian glycans 1618:1634 mammalian glycans with varying avidities 1618:1657 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 3 56 theme new 683:685 arg1 subset 707:712 a new or different glycan subset 681:712 a new or different glycan subset 681:712 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 3 57 theme alternative 581:591 arg1 glycosyltransferase 593:611 an alternative glycosyltransferase 578:611 an alternative glycosyltransferase 578:611 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 7 58 from mutation 1274:1281 arg1 gene 1302:1305 a glycosylation gene 1286:1305 a glycosylation gene 1286:1305 There are now many strategies for generating a precise mutation in a glycosylation gene in a mammalian cell. 24903886 2 59 theme glycosylation 297:309 arg1 mutant 311:316 a mammalian cell glycosylation mutant 280:316 a mammalian cell glycosylation mutant lacking a specific glycosyltransferase 280:355 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 12 60 theme lectins 1987:1993 arg1 assays 1893:1898 rapid assays 1887:1898 rapid assays to compare the cytotoxicity of lectins in a lectin resistance test 1887:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 12 60 theme lectins 1987:1993 arg1 binding 1976:1982 the binding 1972:1982 the binding of lectins or antibodies by flow cytometry 1972:2025 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 0 61 from glycosylation 74:86 arg1 cells 101:105 mammalian cells 91:105 mammalian cells 91:105 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 5 62 theme sugar 937:941 arg1 s 943:943 additional sugar(s) 926:944 additional sugar(s) that are more elaborate than the glycans of wild type cells 926:1004 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 1 63 from complement 219:228 arg1 glycoconjugates 250:264 their glycoconjugates 244:264 their glycoconjugates 244:264 Glycosylation engineering is used to generate glycoproteins, glycolipids, or proteoglycans with a more defined complement of glycans on their glycoconjugates. 24903886 3 64 theme truncated 630:638 arg1 glycans 640:646 the truncated glycans 626:646 the truncated glycans 626:646 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 3 64 theme truncated 630:638 arg1 acceptors 651:659 acceptors 651:659 acceptors 651:659 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 9 65 theme glycosylation 1521:1533 arg1 activity 1535:1542 an altered glycosylation activity 1510:1542 an altered glycosylation activity 1510:1542 This article will focus on how to rapidly characterize mammalian cells with an altered glycosylation activity. 24903886 6 66 theme several 1185:1191 arg1 types 1193:1197 several types 1185:1197 several types of glycoconjugates 1185:1216 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 6 67 from Mutations 1007:1015 arg1 transporters 1097:1108 transporters 1097:1108 transporters 1097:1108 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 6 67 from Mutations 1007:1015 arg1 synthases 1084:1092 nucleotide sugar synthases 1067:1092 nucleotide sugar synthases 1067:1092 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 6 67 from Mutations 1007:1015 arg1 genes 1026:1030 other genes 1020:1030 other genes 1020:1030 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 0 68 theme mammalian 91:99 arg1 cells 101:105 mammalian cells 91:105 mammalian cells 91:105 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 12 69 theme resistance 1951:1960 arg1 test 1962:1965 a lectin resistance test 1942:1965 a lectin resistance test 1942:1965 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 8 70 theme Large-volume 1328:1339 arg1 cultures 1341:1348 Large-volume cultures 1328:1348 Large-volume cultures of mammalian cells 1328:1367 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 6 71 theme sugar 1078:1082 arg1 synthases 1084:1092 nucleotide sugar synthases 1067:1092 nucleotide sugar synthases 1067:1092 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 12 72 theme lectins 1931:1937 arg1 cytotoxicity 1915:1926 the cytotoxicity 1911:1926 the cytotoxicity of lectins 1911:1937 Here we describe rapid assays to compare the cytotoxicity of lectins in a lectin resistance test, and the binding of lectins or antibodies by flow cytometry in a glycan-binding assay. 24903886 8 73 theme cells 1363:1367 arg1 cultures 1341:1348 Large-volume cultures 1328:1348 Large-volume cultures of mammalian cells 1328:1367 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 0 74 theme lectin 17:22 arg1 toxicity 24:31 lectin toxicity 17:31 lectin toxicity 17:31 Rapid assays for lectin toxicity and binding changes that reflect altered glycosylation in mammalian cells. 24903886 13 75 theme gain-of-function 2234:2249 arg1 mutation 2251:2258 a loss-of-function or gain-of-function mutation 2212:2258 a loss-of-function or gain-of-function mutation 2212:2258 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 4 76 theme mutant 764:769 arg1 type 742:745 Another type 734:745 Another type of glycosylation mutant 734:769 Another type of glycosylation mutant arises from gain-of-function mutations that, for example, activate a silent glycosyltransferase gene. 24903886 8 77 theme spontaneous 1387:1397 arg1 mutants 1399:1405 spontaneous mutants 1387:1405 spontaneous mutants in glycosylation pathways 1387:1431 Large-volume cultures of mammalian cells may also generate spontaneous mutants in glycosylation pathways. 24903886 10 78 theme specific 1677:1684 arg1 structure 1686:1694 the specific structure 1673:1694 the specific structure of those glycans 1673:1711 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 6 79 theme glycan 1126:1131 arg1 complement 1133:1142 the glycan complement 1122:1142 the glycan complement in more general ways that usually affect several types of glycoconjugates 1122:1216 Mutations in other genes that affect glycosylation, such as nucleotide sugar synthases or transporters, will alter the glycan complement in more general ways that usually affect several types of glycoconjugates. 24903886 3 80 theme different 690:698 arg1 subset 707:712 a new or different glycan subset 681:712 a new or different glycan subset 681:712 In some cases, an alternative glycosyltransferase may then use the truncated glycans as acceptors, thereby generating a new or different glycan subset in the mutant cell. 24903886 4 81 theme silent 840:845 arg1 gene 867:870 a silent glycosyltransferase gene 838:870 a silent glycosyltransferase gene 838:870 Another type of glycosylation mutant arises from gain-of-function mutations that, for example, activate a silent glycosyltransferase gene. 24903886 13 82 theme loss-of-function 2214:2229 arg1 mutation 2251:2258 a loss-of-function or gain-of-function mutation 2212:2258 a loss-of-function or gain-of-function mutation 2212:2258 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 2 83 with glycoproteins 367:379 arg1 glycans 438:444 truncated glycans 428:444 truncated glycans missing the sugar transferred by that glycosyltransferase 428:502 For example, a mammalian cell glycosylation mutant lacking a specific glycosyltransferase generates glycoproteins, and/or glycolipids, and/or proteoglycans with truncated glycans missing the sugar transferred by that glycosyltransferase, as well as those sugars that would be added subsequently. 24903886 5 84 theme wild 990:993 arg1 cells 1000:1004 wild type cells 990:1004 wild type cells 990:1004 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 11 85 theme altered 1725:1731 arg1 glycosylation 1733:1745 altered glycosylation 1725:1745 altered glycosylation 1725:1745 Cells with altered glycosylation generally become resistant or hypersensitive to lectin toxicity, and have reduced or increased lectin or antibody binding. 24903886 10 86 theme glycans 1705:1711 arg1 structure 1686:1694 the specific structure 1673:1694 the specific structure of those glycans 1673:1711 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 24903886 13 87 theme phenotypic 2177:2186 arg1 groups 2188:2193 phenotypic groups 2177:2193 phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis 2177:2306 Based on these tests, glycosylation changes expressed by a cell can be revealed, and glycosylation mutants classified into phenotypic groups that may reflect a loss-of-function or gain-of-function mutation in a specific gene involved in glycan synthesis. 24903886 9 88 theme mammalian 1489:1497 arg1 cells 1499:1503 mammalian cells 1489:1503 mammalian cells with an altered glycosylation activity 1489:1542 This article will focus on how to rapidly characterize mammalian cells with an altered glycosylation activity. 24903886 5 89 theme cells 1000:1004 arg1 glycans 979:985 the glycans 975:985 the glycans of wild type cells 975:1004 In this case, glycoconjugates will have glycans with additional sugar(s) that are more elaborate than the glycans of wild type cells. 24903886 10 90 with glycans 1628:1634 arg1 avidities 1649:1657 varying avidities 1641:1657 varying avidities 1641:1657 The key reagents for the protocols described are plant lectins that bind mammalian glycans with varying avidities, depending on the specific structure of those glycans. 25793407 6 0 theme patients 1036:1043 arg1 sera 1023:1026 the sera 1019:1026 the sera of T2DM patients 1019:1043 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 2 1 theme Many 343:346 arg1 roles 369:373 Many important biological roles 343:373 Many important biological roles of glycoproteins 343:390 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 4 2 theme healthy 846:852 arg1 CTRs 872:875 CTRs 872:875 CTRs 872:875 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 2 theme healthy 846:852 arg1 subjects 862:869 599 healthy control subjects 842:869 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years) 842:904 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 3 3 theme diabetes 575:582 arg1 T2DM 594:597 T2DM 594:597 T2DM 594:597 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 3 theme diabetes 575:582 arg1 mellitus 584:591 type 2 diabetes mellitus 568:591 type 2 diabetes mellitus (T2DM) 568:598 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 11 4 dep T2DM 1679:1682 arg1 compared 1687:1694 compared 1687:1694 compared to healthy controls 1687:1714 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 5 theme " 1801:1801 arg1 MS 1767:1768 MS 1767:1768 MS 1767:1768 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 5 theme " 1801:1801 arg1 complications 1749:1761 complications 1749:1761 complications 1749:1761 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 5 theme " 1801:1801 arg1 phenotype 1803:1811 the extreme "unhealthy" phenotype 1779:1811 the extreme "unhealthy" phenotype (T2DM+ with MS) 1779:1827 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 4 6 theme age 818:820 arg1 years 831:835 mean age 65.6±8.2 years 813:835 mean age 65.6±8.2 years 813:835 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 6 theme age 818:820 arg1 patients 766:773 the study 562 patients 752:773 the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) 752:836 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 2 7 theme glycoproteins 378:390 arg1 roles 369:373 Many important biological roles 343:373 Many important biological roles of glycoproteins 343:390 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 12 8 theme levels 1917:1922 arg1 Imbalance 1842:1850 CONCLUSIONS Imbalance 1830:1850 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels 1830:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 3 9 theme serum 712:716 arg1 N-glycome 718:726 serum N-glycome 712:726 serum N-glycome 712:726 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 7 10 theme diantennary 1116:1126 arg1 N-glycans 1128:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans 1061:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) 1061:1148 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 7 10 theme diantennary 1116:1126 arg1 A2F 1145:1147 NG1(6)A2F 1139:1147 NG1(6)A2F 1139:1147 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 10 11 theme most 1603:1606 arg1 parameters 1611:1620 most MS parameters 1603:1620 most MS parameters 1603:1620 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 1 12 theme oligosaccharides 190:205 arg1 addition 178:185 the enzymatic addition 164:185 the enzymatic addition 164:185 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 10 13 theme NG1 1451:1453 arg1 A2F 1457:1459 NG1(3)A2F 1451:1459 NG1(3)A2F 1451:1459 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 0 14 theme syndrome 113:120 arg1 parameters 122:131 metabolic syndrome parameters 103:131 metabolic syndrome parameters 103:131 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 12 15 theme nucleotide 1900:1909 arg1 levels 1917:1922 sugar nucleotide donor levels 1894:1922 sugar nucleotide donor levels 1894:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 11 16 with patients 1735:1742 arg1 MS 1767:1768 MS 1767:1768 MS 1767:1768 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 16 with patients 1735:1742 arg1 complications 1749:1761 complications 1749:1761 complications 1749:1761 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 16 with patients 1735:1742 arg1 phenotype 1803:1811 the extreme "unhealthy" phenotype 1779:1811 the extreme "unhealthy" phenotype (T2DM+ with MS) 1779:1827 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 0 17 theme serum 22:26 arg1 proteins 28:35 serum proteins 22:35 serum proteins 22:35 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 9 18 theme NG1 1413:1415 arg1 A2F 1419:1421 NG1(6)A2F 1413:1421 NG1(6)A2F 1413:1421 Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. 25793407 1 19 theme BACKGROUND 134:143 arg1 Glycosylation 145:157 BACKGROUND Glycosylation 134:157 BACKGROUND Glycosylation 134:157 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 0 20 theme diabetes 47:54 arg1 mellitus 56:63 type 2 diabetes mellitus 40:63 type 2 diabetes mellitus 40:63 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 1 21 theme modifications 316:328 arg1 one 271:273 one 271:273 one 271:273 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 1 21 theme modifications 316:328 arg1 modifications 316:328 the most common co-/posttranslational modifications 278:328 the most common co-/posttranslational modifications of proteins 278:340 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 9 22 theme A2F 1419:1421 arg1 levels 1403:1408 decreased levels 1393:1408 decreased levels of NG1(6)A2F 1393:1421 Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. 25793407 7 23 dep monogalactosylated 1079:1096 arg1 core-fucosylated 1099:1114 core-fucosylated 1099:1114 core-fucosylated 1099:1114 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 12 24 theme CONCLUSIONS 1830:1840 arg1 Imbalance 1842:1850 CONCLUSIONS Imbalance 1830:1850 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels 1830:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 1 25 theme proteins 333:340 arg1 modifications 316:328 the most common co-/posttranslational modifications 278:328 the most common co-/posttranslational modifications of proteins 278:340 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 4 26 theme 562 762:764 arg1 years 831:835 mean age 65.6±8.2 years 813:835 mean age 65.6±8.2 years 813:835 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 26 theme 562 762:764 arg1 patients 766:773 the study 562 patients 752:773 the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) 752:836 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 13 27 theme Serum 2070:2074 arg1 levels 2085:2090 MS. Serum N-glycan levels 2066:2090 MS. Serum N-glycan levels 2066:2090 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 6 28 dep RESULTS 955:961 arg1 found 966:970 found 966:970 found significant changes in N-glycan composition in the sera of T2DM patients 966:1043 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 10 29 dep -antennary 1541:1550 arg1 α 1523:1523 α 1523:1523 α 1523:1523 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 10 29 dep -antennary 1541:1550 arg1 α 1535:1535 α 1535:1535 α 1535:1535 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 10 29 dep -antennary 1541:1550 arg1 1,6 1525:1527 1,6 1525:1527 1,6 1525:1527 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 11 30 theme healthy 1699:1705 arg1 controls 1707:1714 healthy controls 1699:1714 healthy controls 1699:1714 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 0 31 theme N-glycomic 0:9 arg1 changes 11:17 N-glycomic changes 0:17 N-glycomic changes in serum proteins in type 2 diabetes mellitus 0:63 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 3 32 gly glycosylation 489:501 arg1 proteins 506:513 proteins 506:513 proteins 506:513 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 11 33 theme extreme 1783:1789 arg1 MS 1767:1768 MS 1767:1768 MS 1767:1768 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 33 theme extreme 1783:1789 arg1 complications 1749:1761 complications 1749:1761 complications 1749:1761 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 33 theme extreme 1783:1789 arg1 phenotype 1803:1811 the extreme "unhealthy" phenotype 1779:1811 the extreme "unhealthy" phenotype (T2DM+ with MS) 1779:1827 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 4 34 with subjects 862:869 arg1 T2DM 806:809 T2DM 806:809 T2DM 806:809 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 34 with subjects 862:869 arg1 Mellitus 796:803 Type 2 Diabetes Mellitus 780:803 Type 2 Diabetes Mellitus (T2DM) 780:810 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 6 35 theme N-glycan 995:1002 arg1 composition 1004:1014 N-glycan composition 995:1014 N-glycan composition 995:1014 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 9 36 with related 1380:1386 arg1 levels 1403:1408 decreased levels 1393:1408 decreased levels of NG1(6)A2F 1393:1421 Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. 25793407 3 37 theme glucose 650:656 arg1 levels 658:663 altered glucose levels 642:663 altered glucose levels 642:663 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 2 38 theme N-linked 409:416 arg1 oligosaccharides 418:433 N-linked oligosaccharides 409:433 N-linked oligosaccharides 409:433 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 0 39 theme type 40:43 arg1 mellitus 56:63 type 2 diabetes mellitus 40:63 type 2 diabetes mellitus 40:63 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 1 40 theme common 287:292 arg1 modifications 316:328 the most common co-/posttranslational modifications 278:328 the most common co-/posttranslational modifications of proteins 278:340 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 7 41 theme CTR 1199:1201 arg1 subjects 1203:1210 CTR subjects 1199:1210 CTR subjects 1199:1210 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 11 42 theme glycans 1657:1663 arg1 lower 1670:1674 lower 1670:1674 lower 1670:1674 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 42 theme glycans 1657:1663 arg1 levels 1637:1642 The plasmatic levels 1623:1642 The plasmatic levels of these two glycans 1623:1663 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 5 43 theme serum 934:938 arg1 glycoproteins 940:952 serum glycoproteins 934:952 serum glycoproteins 934:952 N-glycome was evaluated in serum glycoproteins. 25793407 12 44 theme structural 1945:1954 arg1 changes 1956:1962 the structural changes 1941:1962 the structural changes evidenced by our findings 1941:1988 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 10 45 theme -antennary 1541:1550 arg1 galactosylation 1552:1566 α(1,6)- and α(1,3)-antennary galactosylation 1523:1566 α(1,6)- and α(1,3)-antennary galactosylation 1523:1566 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 2 46 theme important 348:356 arg1 roles 369:373 Many important biological roles 343:373 Many important biological roles of glycoproteins 343:390 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 6 47 theme T2DM 1031:1034 arg1 patients 1036:1043 T2DM patients 1031:1043 T2DM patients 1031:1043 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 3 48 theme specific 686:693 arg1 modifications 695:707 specific modifications 686:707 specific modifications in serum N-glycome 686:726 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 11 49 theme unhealthy 1792:1800 arg1 MS 1767:1768 MS 1767:1768 MS 1767:1768 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 49 theme unhealthy 1792:1800 arg1 complications 1749:1761 complications 1749:1761 complications 1749:1761 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 49 theme unhealthy 1792:1800 arg1 phenotype 1803:1811 the extreme "unhealthy" phenotype 1779:1811 the extreme "unhealthy" phenotype (T2DM+ with MS) 1779:1827 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 9 50 theme Macro 1334:1338 arg1 vascular-complications 1340:1361 Macro vascular-complications 1334:1361 Macro vascular-complications 1334:1361 Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. 25793407 3 51 theme type 568:571 arg1 T2DM 594:597 T2DM 594:597 T2DM 594:597 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 51 theme type 568:571 arg1 mellitus 584:591 type 2 diabetes mellitus 568:591 type 2 diabetes mellitus (T2DM) 568:598 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 52 theme metabolic 540:548 arg1 MS 560:561 MS 560:561 MS 560:561 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 52 theme metabolic 540:548 arg1 syndrome 550:557 metabolic syndrome 540:557 metabolic syndrome (MS) 540:562 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 52 theme metabolic 540:548 arg1 conditions 614:623 pathological conditions 601:623 pathological conditions characterized by altered glucose levels 601:663 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 1 53 theme enzymatic 168:176 arg1 addition 178:185 the enzymatic addition 164:185 the enzymatic addition 164:185 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 13 54 theme Serum 1991:1995 arg1 profiles 2006:2013 Serum N-glycan profiles 1991:2013 Serum N-glycan profiles 1991:2013 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 4 55 theme mean 813:816 arg1 years 831:835 mean age 65.6±8.2 years 813:835 mean age 65.6±8.2 years 813:835 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 55 theme mean 813:816 arg1 patients 766:773 the study 562 patients 752:773 the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) 752:836 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 12 56 theme donor 1911:1915 arg1 levels 1917:1922 sugar nucleotide donor levels 1894:1922 sugar nucleotide donor levels 1894:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 6 57 from changes 984:990 arg1 composition 1004:1014 N-glycan composition 995:1014 N-glycan composition 995:1014 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 10 58 theme MS 1608:1609 arg1 parameters 1611:1620 most MS parameters 1603:1620 most MS parameters 1603:1620 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 13 59 theme non-invasive 2118:2129 arg1 marker 2143:2148 a non-invasive alternative marker 2116:2148 a non-invasive alternative marker for T2DM and MS 2116:2164 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 4 60 with patients 766:773 arg1 T2DM 806:809 T2DM 806:809 T2DM 806:809 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 60 with patients 766:773 arg1 Mellitus 796:803 Type 2 Diabetes Mellitus 780:803 Type 2 Diabetes Mellitus (T2DM) 780:810 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 13 61 theme N-glycan 2076:2083 arg1 levels 2085:2090 MS. Serum N-glycan levels 2066:2090 MS. Serum N-glycan levels 2066:2090 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 12 62 theme glycosidases 1877:1888 arg1 Imbalance 1842:1850 CONCLUSIONS Imbalance 1830:1850 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels 1830:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 13 63 theme alternative 2131:2141 arg1 marker 2143:2148 a non-invasive alternative marker 2116:2148 a non-invasive alternative marker for T2DM and MS 2116:2164 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 9 64 theme decreased 1393:1401 arg1 levels 1403:1408 decreased levels 1393:1408 decreased levels of NG1(6)A2F 1393:1421 Macro vascular-complications were found to be related with decreased levels of NG1(6)A2F. 25793407 4 65 theme mean 879:882 arg1 years 899:903 58.5±12.4 years 889:903 58.5±12.4 years 889:903 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 65 theme mean 879:882 arg1 age 884:886 mean age 879:886 mean age 879:886 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 12 66 theme sugar 1894:1898 arg1 levels 1917:1922 sugar nucleotide donor levels 1894:1922 sugar nucleotide donor levels 1894:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 3 67 from modifications 695:707 arg1 N-glycome 718:726 serum N-glycome 712:726 serum N-glycome 712:726 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 11 68 theme plasmatic 1627:1635 arg1 lower 1670:1674 lower 1670:1674 lower 1670:1674 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 11 68 theme plasmatic 1627:1635 arg1 levels 1637:1642 The plasmatic levels 1623:1642 The plasmatic levels of these two glycans 1623:1663 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 7 69 link -linked 1067:1073 arg1 N-glycans 1128:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans 1061:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) 1061:1148 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 7 69 link -linked 1067:1073 arg1 A2F 1145:1147 NG1(6)A2F 1139:1147 NG1(6)A2F 1139:1147 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 13 70 theme diabetes 2053:2060 arg1 presence 2041:2048 the presence 2037:2048 the presence of diabetes 2037:2060 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 4 71 theme control 854:860 arg1 CTRs 872:875 CTRs 872:875 CTRs 872:875 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 71 theme control 854:860 arg1 subjects 862:869 599 healthy control subjects 842:869 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years) 842:904 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 7 72 theme monogalactosylated 1079:1096 arg1 N-glycans 1128:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans 1061:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) 1061:1148 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 7 72 theme monogalactosylated 1079:1096 arg1 A2F 1145:1147 NG1(6)A2F 1139:1147 NG1(6)A2F 1139:1147 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 3 73 theme pathological 601:612 arg1 syndrome 550:557 metabolic syndrome 540:557 metabolic syndrome (MS) 540:562 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 3 73 theme pathological 601:612 arg1 conditions 614:623 pathological conditions 601:623 pathological conditions characterized by altered glucose levels 601:663 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 13 74 theme MS. 2066:2068 arg1 levels 2085:2090 MS. Serum N-glycan levels 2066:2090 MS. Serum N-glycan levels 2066:2090 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 6 75 theme significant 972:982 arg1 changes 984:990 significant changes 972:990 significant changes in N-glycan composition 972:1014 RESULTS We found significant changes in N-glycan composition in the sera of T2DM patients. 25793407 8 76 theme diabetic 1255:1262 arg1 patients 1264:1271 diabetic patients 1255:1271 diabetic patients with and without complications (P<0.001) 1255:1312 Importantly, they were equally reduced in diabetic patients with and without complications (P<0.001) compared with CTRs. 25793407 12 77 theme glycosyltransferases 1855:1874 arg1 Imbalance 1842:1850 CONCLUSIONS Imbalance 1830:1850 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels 1830:1922 CONCLUSIONS Imbalance of glycosyltransferases, glycosidases and sugar nucleotide donor levels is able to cause the structural changes evidenced by our findings. 25793407 11 78 dep phenotype 1803:1811 arg1 T2DM+ 1814:1818 T2DM+ 1814:1818 T2DM+ 1814:1818 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 4 79 dep subjects 862:869 arg1 years 899:903 58.5±12.4 years 889:903 58.5±12.4 years 889:903 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 79 dep subjects 862:869 arg1 age 884:886 mean age 879:886 mean age 879:886 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 0 80 from changes 11:17 arg1 mellitus 56:63 type 2 diabetes mellitus 40:63 type 2 diabetes mellitus 40:63 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 0 80 from changes 11:17 arg1 proteins 28:35 serum proteins 22:35 serum proteins 22:35 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 10 81 dep resulted 1569:1576 arg1 correlated 1587:1596 correlated 1587:1596 resulted strongly correlated with most MS parameters 1569:1620 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 7 82 theme NG1 1139:1141 arg1 N-glycans 1128:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans 1061:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) 1061:1148 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 7 82 theme NG1 1139:1141 arg1 A2F 1145:1147 NG1(6)A2F 1139:1147 NG1(6)A2F 1139:1147 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 4 83 theme study 756:760 arg1 years 831:835 mean age 65.6±8.2 years 813:835 mean age 65.6±8.2 years 813:835 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 83 theme study 756:760 arg1 patients 766:773 the study 562 patients 752:773 the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) 752:836 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 0 84 theme metabolic 103:111 arg1 parameters 122:131 metabolic syndrome parameters 103:131 metabolic syndrome parameters 103:131 N-glycomic changes in serum proteins in type 2 diabetes mellitus correlate with complications and with metabolic syndrome parameters. 25793407 10 85 theme monogalactosylated 1489:1506 arg1 N-glycans 1508:1516 monogalactosylated N-glycans 1489:1516 monogalactosylated N-glycans 1489:1516 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 4 86 dep METHODS 729:735 arg1 enrolled 740:747 enrolled 740:747 enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years) 740:904 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 2 87 gly glycoproteins 378:390 arg1 glycoproteins 378:390 glycoproteins 378:390 glycoproteins 378:390 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 3 88 theme glucose 439:445 arg1 levels 447:452 glucose levels 439:452 glucose levels 439:452 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 11 89 with T2DM+ 1814:1818 arg1 MS 1825:1826 MS 1825:1826 MS 1825:1826 The plasmatic levels of these two glycans were lower in T2DM as compared to healthy controls, and even lower in patients with complications and MS, that is the extreme "unhealthy" phenotype (T2DM+ with MS). 25793407 13 90 theme N-glycan 1997:2004 arg1 profiles 2006:2013 Serum N-glycan profiles 1991:2013 Serum N-glycan profiles 1991:2013 Serum N-glycan profiles are thus sensitive to the presence of diabetes and MS. Serum N-glycan levels could therefore provide a non-invasive alternative marker for T2DM and MS. 25793407 3 91 theme altered 642:648 arg1 levels 658:663 altered glucose levels 642:663 altered glucose levels 642:663 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 7 92 theme -linked 1067:1073 arg1 N-glycans 1128:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans 1061:1136 α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) 1061:1148 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 7 92 theme -linked 1067:1073 arg1 A2F 1145:1147 NG1(6)A2F 1139:1147 NG1(6)A2F 1139:1147 In particular, α(1,6)-linked arm monogalactosylated, core-fucosylated diantennary N-glycans (NG1(6)A2F) were significantly reduced in T2DM compared with CTR subjects. 25793407 1 93 dep oligosaccharides 190:205 arg1 glycans 211:217 glycans 211:217 glycans 211:217 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 5 94 gly glycoproteins 940:952 arg1 glycoproteins 940:952 serum glycoproteins 934:952 serum glycoproteins 934:952 N-glycome was evaluated in serum glycoproteins. 25793407 2 95 link N-linked 409:416 arg1 oligosaccharides 418:433 N-linked oligosaccharides 409:433 N-linked oligosaccharides 409:433 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25793407 3 96 theme proteins 506:513 arg1 glycosylation 489:501 glycosylation 489:501 glycosylation of proteins 489:513 As glucose levels can affect the pathways leading to glycosylation of proteins, we investigated whether metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM), pathological conditions characterized by altered glucose levels, are associated with specific modifications in serum N-glycome. 25793407 1 97 theme co-/posttranslational 294:314 arg1 modifications 316:328 the most common co-/posttranslational modifications 278:328 the most common co-/posttranslational modifications of proteins 278:340 BACKGROUND Glycosylation, i.e the enzymatic addition of oligosaccharides (or glycans) to proteins and lipids, known as glycosylation, is one of the most common co-/posttranslational modifications of proteins. 25793407 4 98 theme Type 780:783 arg1 T2DM 806:809 T2DM 806:809 T2DM 806:809 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 98 theme Type 780:783 arg1 Mellitus 796:803 Type 2 Diabetes Mellitus 780:803 Type 2 Diabetes Mellitus (T2DM) 780:810 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 10 99 theme NG1 1437:1439 arg1 A2F 1443:1445 NG1(6)A2F 1437:1445 NG1(6)A2F 1437:1445 In addition, NG1(6)A2F and NG1(3)A2F, identifying, respectively, monogalactosylated N-glycans with α(1,6)- and α(1,3)-antennary galactosylation, resulted strongly correlated with most MS parameters. 25793407 4 100 theme Diabetes 787:794 arg1 T2DM 806:809 T2DM 806:809 T2DM 806:809 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 4 100 theme Diabetes 787:794 arg1 Mellitus 796:803 Type 2 Diabetes Mellitus 780:803 Type 2 Diabetes Mellitus (T2DM) 780:810 METHODS We enrolled in the study 562 patients with Type 2 Diabetes Mellitus (T2DM) (mean age 65.6±8.2 years) and 599 healthy control subjects (CTRs) (mean age, 58.5±12.4 years). 25793407 2 101 theme biological 358:367 arg1 roles 369:373 Many important biological roles 343:373 Many important biological roles of glycoproteins 343:390 Many important biological roles of glycoproteins are modulated by N-linked oligosaccharides. 25479762 6 0 theme hGGT1 1101:1105 arg1 content 1090:1096 N-glycan content 1081:1096 N-glycan content of hGGT1 in complex biological samples 1081:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 1 1 theme γ-glutamyl-containing 275:295 arg1 substrates 297:306 other γ-glutamyl-containing substrates 269:306 other γ-glutamyl-containing substrates 269:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 12 2 theme ALSA 2058:2061 arg1 implementation 2040:2053 the implementation 2036:2053 the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2036:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 6 3 from content 1090:1096 arg1 samples 1129:1135 complex biological samples 1110:1135 complex biological samples 1110:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 12 4 theme conducting 2066:2075 arg1 studies 2093:2099 conducting high-throughput studies 2066:2099 conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2066:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 12 5 theme enhancing 2204:2212 arg1 diagnoses 2223:2231 enhancing clinical diagnoses 2204:2231 enhancing clinical diagnoses 2204:2231 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 8 6 with hGGT1 1271:1275 arg1 lectins 1282:1288 lectins 1282:1288 lectins 1282:1288 Probing hGGT1 with lectins enables characterization of the N-glycans. 25479762 1 7 gly N-glycosylated 190:203 arg1 transpeptidase 160:173 BACKGROUND γ-Glutamyl transpeptidase 1 138:175 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) 138:182 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 7 gly N-glycosylated 190:203 arg1 protein 214:220 an N-glycosylated membrane protein 187:220 an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates 187:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 3 8 from N-glycans 468:476 arg1 hGGT1 493:497 hGGT1 493:497 hGGT1 493:497 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 8 from N-glycans 468:476 arg1 GGT1 487:490 human GGT1 481:490 human GGT1 (hGGT1) 481:498 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 9 9 theme yeast 1436:1440 arg1 pastoris 1449:1456 the yeast Pichia pastoris 1432:1456 the yeast Pichia pastoris 1432:1456 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 10 10 from sources 1649:1655 arg1 characterizations 1620:1636 previous large-scale hGGT1 N-glycan characterizations 1584:1636 previous large-scale hGGT1 N-glycan characterizations from these sources 1584:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 12 11 theme diagnoses 2223:2231 arg1 goal 2196:2199 the goal 2192:2199 the goal of enhancing clinical diagnoses and targeted treatment regimens 2192:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 10 12 theme N-glycan 1611:1618 arg1 characterizations 1620:1636 previous large-scale hGGT1 N-glycan characterizations 1584:1636 previous large-scale hGGT1 N-glycan characterizations from these sources 1584:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 11 13 theme high-mannose-type 1835:1851 arg1 N-glycans 1853:1861 high-mannose-type N-glycans 1835:1861 high-mannose-type N-glycans 1835:1861 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 4 14 from N-glycans 622:630 arg1 hGGT1 635:639 hGGT1 635:639 hGGT1 635:639 Tumor-specific changes in the glycans have also been observed, suggesting that the N-glycans on hGGT1 would be an important biomarker for detecting tumors and monitoring their progression during treatment. 25479762 6 15 theme ALSA 970:973 arg1 platform 976:983 a new antibody-lectin sandwich array (ALSA) platform 932:983 a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples 932:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 10 16 theme large-scale 1593:1603 arg1 characterizations 1620:1636 previous large-scale hGGT1 N-glycan characterizations 1584:1636 previous large-scale hGGT1 N-glycan characterizations from these sources 1584:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 2 17 theme disease 325:331 arg1 states 333:338 disease states 325:338 disease states 325:338 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 1 18 theme membrane 205:212 arg1 transpeptidase 160:173 BACKGROUND γ-Glutamyl transpeptidase 1 138:175 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) 138:182 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 18 theme membrane 205:212 arg1 protein 214:220 an N-glycosylated membrane protein 187:220 an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates 187:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 6 19 theme array 963:967 arg1 platform 976:983 a new antibody-lectin sandwich array (ALSA) platform 932:983 a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples 932:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 6 20 theme antibody-lectin 938:952 arg1 platform 976:983 a new antibody-lectin sandwich array (ALSA) platform 932:983 a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples 932:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 12 21 theme glycosylation 2155:2167 arg1 patterns 2169:2176 the glycosylation patterns 2151:2176 the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2151:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 11 22 theme refined 1773:1779 arg1 evidence 1781:1788 refined evidence 1773:1788 refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors 1773:1953 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 1 23 theme BACKGROUND 138:147 arg1 transpeptidase 160:173 BACKGROUND γ-Glutamyl transpeptidase 1 138:175 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) 138:182 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 23 theme BACKGROUND 138:147 arg1 GGT1 178:181 GGT1 178:181 GGT1 178:181 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 23 theme BACKGROUND 138:147 arg1 protein 214:220 an N-glycosylated membrane protein 187:220 an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates 187:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 4 24 theme important 653:661 arg1 biomarker 663:671 an important biomarker 650:671 an important biomarker for detecting tumors and monitoring their progression during treatment 650:742 Tumor-specific changes in the glycans have also been observed, suggesting that the N-glycans on hGGT1 would be an important biomarker for detecting tumors and monitoring their progression during treatment. 25479762 4 24 theme important 653:661 arg1 N-glycans 622:630 the N-glycans 618:630 the N-glycans on hGGT1 618:639 Tumor-specific changes in the glycans have also been observed, suggesting that the N-glycans on hGGT1 would be an important biomarker for detecting tumors and monitoring their progression during treatment. 25479762 5 25 theme biomarker 885:893 arg1 development 895:905 biomarker development 885:905 biomarker development 885:905 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 10 26 with consistent 1523:1532 arg1 composition 1558:1568 the hGGT1 N-glycan composition 1539:1568 the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources 1539:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 0 27 from Detection 0:8 arg1 transpeptidase 65:78 human γ-glutamyl transpeptidase 1 48:80 human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology 48:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 0 28 theme sandwich 104:111 arg1 technology 126:135 antibody-lectin sandwich array (ALSA) technology 88:135 antibody-lectin sandwich array (ALSA) technology 88:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 10 29 theme binding 1470:1476 arg1 patterns 1478:1485 The lectin binding patterns 1459:1485 The lectin binding patterns obtained with the ALSA platform 1459:1517 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 10 29 theme binding 1470:1476 arg1 consistent 1523:1532 consistent 1523:1532 consistent 1523:1532 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 6 30 theme N-glycan 1081:1088 arg1 content 1090:1096 N-glycan content 1081:1096 N-glycan content of hGGT1 in complex biological samples 1081:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 11 31 from implementation 1679:1692 arg1 platform 1752:1759 this platform 1747:1759 this platform 1747:1759 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 0 32 theme distinct 13:20 arg1 patterns 36:43 distinct glycosylation patterns 13:43 distinct glycosylation patterns 13:43 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 9 33 theme normal 1354:1359 arg1 tissue 1373:1378 normal human liver tissue 1354:1378 normal human liver tissue 1354:1378 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 12 34 from implementation 2040:2053 arg1 studies 2093:2099 conducting high-throughput studies 2066:2099 conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2066:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 12 35 theme disease-related 2124:2138 arg1 changes 2140:2146 disease-related changes 2124:2146 disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2124:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 4 36 theme Tumor-specific 539:552 arg1 changes 554:560 Tumor-specific changes 539:560 Tumor-specific changes in the glycans 539:575 Tumor-specific changes in the glycans have also been observed, suggesting that the N-glycans on hGGT1 would be an important biomarker for detecting tumors and monitoring their progression during treatment. 25479762 0 37 theme patterns 36:43 arg1 Detection 0:8 Detection 0:8 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 0:136 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 9 38 theme liver 1367:1371 arg1 tissue 1373:1378 normal human liver tissue 1354:1378 normal human liver tissue 1354:1378 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 11 39 theme lectin 1724:1729 arg1 implementation 1679:1692 the implementation 1675:1692 the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform 1675:1759 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 3 40 from structures 450:459 arg1 hGGT1 493:497 hGGT1 493:497 hGGT1 493:497 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 40 from structures 450:459 arg1 GGT1 487:490 human GGT1 481:490 human GGT1 (hGGT1) 481:498 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 41 theme human 481:485 arg1 hGGT1 493:497 hGGT1 493:497 hGGT1 493:497 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 41 theme human 481:485 arg1 GGT1 487:490 human GGT1 481:490 human GGT1 (hGGT1) 481:498 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 5 42 theme purified 778:785 arg1 protein 787:793 purified protein 778:793 purified protein 778:793 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 12 43 theme targeted 2237:2244 arg1 regimens 2256:2263 targeted treatment regimens 2237:2263 targeted treatment regimens 2237:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 11 44 theme carbohydrate 1875:1886 arg1 modification 1888:1899 carbohydrate modification 1875:1899 carbohydrate modification 1875:1899 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 9 45 theme human 1388:1392 arg1 tissue 1401:1406 normal human kidney tissue 1381:1406 normal human kidney tissue 1381:1406 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 6 46 theme biological 1118:1127 arg1 samples 1129:1135 complex biological samples 1110:1135 complex biological samples 1110:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 10 47 theme N-glycan 1549:1556 arg1 composition 1558:1568 the hGGT1 N-glycan composition 1539:1568 the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources 1539:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 12 48 theme regimens 2256:2263 arg1 goal 2196:2199 the goal 2192:2199 the goal of enhancing clinical diagnoses and targeted treatment regimens 2192:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 11 49 theme hGGT1 1924:1928 arg1 distinctive 1909:1919 distinctive 1909:1919 distinctive 1909:1919 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 12 50 theme high-throughput 2077:2091 arg1 studies 2093:2099 conducting high-throughput studies 2066:2099 conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2066:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 12 51 with hGGT1 2181:2185 arg1 goal 2196:2199 the goal 2192:2199 the goal of enhancing clinical diagnoses and targeted treatment regimens 2192:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 1 52 theme other 269:273 arg1 substrates 297:306 other γ-glutamyl-containing substrates 269:306 other γ-glutamyl-containing substrates 269:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 7 53 theme ALSA 1248:1251 arg1 platform 1253:1260 the ALSA platform 1244:1260 the ALSA platform 1244:1260 RESULTS Our data show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform. 25479762 10 54 theme previous 1584:1591 arg1 characterizations 1620:1636 previous large-scale hGGT1 N-glycan characterizations 1584:1636 previous large-scale hGGT1 N-glycan characterizations from these sources 1584:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 11 55 theme Microcystis 1701:1711 arg1 microvirin 1732:1741 microvirin 1732:1741 microvirin 1732:1741 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 11 55 theme Microcystis 1701:1711 arg1 lectin 1724:1729 the Microcystis aeruginosa lectin 1697:1729 the Microcystis aeruginosa lectin 1697:1729 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 5 56 theme significant 859:869 arg1 challenge 871:879 a significant challenge 857:879 a significant challenge for biomarker development 857:905 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 0 57 theme human 48:52 arg1 transpeptidase 65:78 human γ-glutamyl transpeptidase 1 48:80 human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology 48:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 9 58 theme Pichia 1442:1447 arg1 pastoris 1449:1456 the yeast Pichia pastoris 1432:1456 the yeast Pichia pastoris 1432:1456 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 2 59 located detected 427:434 arg1 serum 439:443 serum 439:443 serum 439:443 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 2 59 located detected 427:434 arg2 enzyme 372:377 the enzyme 368:377 the enzyme 368:377 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 12 60 theme clinical 2214:2221 arg1 diagnoses 2223:2231 enhancing clinical diagnoses 2204:2231 enhancing clinical diagnoses 2204:2231 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 10 61 theme hGGT1 1605:1609 arg1 characterizations 1620:1636 previous large-scale hGGT1 N-glycan characterizations 1584:1636 previous large-scale hGGT1 N-glycan characterizations from these sources 1584:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 3 62 from GGT1 487:490 arg1 structures 450:459 The structures 446:459 The structures of the N-glycans on human GGT1 (hGGT1) 446:498 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 62 from GGT1 487:490 arg1 tissue-specific 522:536 tissue-specific 522:536 tissue-specific 522:536 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 2 63 theme states 333:338 arg1 variety 314:320 a variety 312:320 a variety 312:320 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 2 63 theme states 333:338 arg1 states 333:338 disease states 325:338 disease states 325:338 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 2 63 theme states 333:338 arg1 formation 357:365 tumor formation 351:365 tumor formation 351:365 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 1 64 theme N-glycosylated 190:203 arg1 transpeptidase 160:173 BACKGROUND γ-Glutamyl transpeptidase 1 138:175 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) 138:182 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 64 theme N-glycosylated 190:203 arg1 protein 214:220 an N-glycosylated membrane protein 187:220 an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates 187:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 8 65 theme Probing 1263:1269 arg1 hGGT1 1271:1275 Probing hGGT1 1263:1275 Probing hGGT1 with lectins 1263:1288 Probing hGGT1 with lectins enables characterization of the N-glycans. 25479762 5 66 theme carbohydrate 830:841 arg1 content 843:849 the carbohydrate content 826:849 the carbohydrate content poses a significant challenge for biomarker development 826:905 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 6 67 theme sandwich 954:961 arg1 platform 976:983 a new antibody-lectin sandwich array (ALSA) platform 932:983 a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples 932:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 11 68 dep Microcystis 1701:1711 arg1 aeruginosa 1713:1722 aeruginosa 1713:1722 aeruginosa 1713:1722 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 12 69 from patterns 2169:2176 arg1 hGGT1 2181:2185 hGGT1 2181:2185 hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2181:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 6 70 theme new 934:936 arg1 platform 976:983 a new antibody-lectin sandwich array (ALSA) platform 932:983 a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples 932:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 3 71 theme N-glycans 468:476 arg1 structures 450:459 The structures 446:459 The structures of the N-glycans on human GGT1 (hGGT1) 446:498 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 3 71 theme N-glycans 468:476 arg1 tissue-specific 522:536 tissue-specific 522:536 tissue-specific 522:536 The structures of the N-glycans on human GGT1 (hGGT1) have been shown to be tissue-specific. 25479762 12 72 from changes 2140:2146 arg1 hGGT1 2181:2185 hGGT1 2181:2185 hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2181:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 12 72 from changes 2140:2146 arg1 patterns 2169:2176 the glycosylation patterns 2151:2176 the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2151:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 1 73 theme γ-Glutamyl 149:158 arg1 transpeptidase 160:173 BACKGROUND γ-Glutamyl transpeptidase 1 138:175 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) 138:182 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 73 theme γ-Glutamyl 149:158 arg1 GGT1 178:181 GGT1 178:181 GGT1 178:181 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 1 73 theme γ-Glutamyl 149:158 arg1 protein 214:220 an N-glycosylated membrane protein 187:220 an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates 187:306 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 6 74 theme microanalytical 1011:1025 arg1 technique 1027:1035 this microanalytical technique 1006:1035 this microanalytical technique 1006:1035 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 7 75 dep RESULTS 1138:1144 arg1 show 1155:1158 show 1155:1158 show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform 1155:1260 RESULTS Our data show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform. 25479762 8 76 theme N-glycans 1322:1330 arg1 characterization 1298:1313 characterization 1298:1313 characterization of the N-glycans 1298:1330 Probing hGGT1 with lectins enables characterization of the N-glycans. 25479762 7 77 theme membrane 1212:1219 arg1 proteins 1221:1228 membrane proteins 1212:1228 membrane proteins 1212:1228 RESULTS Our data show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform. 25479762 0 78 theme antibody-lectin 88:102 arg1 technology 126:135 antibody-lectin sandwich array (ALSA) technology 88:135 antibody-lectin sandwich array (ALSA) technology 88:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 0 79 theme array 113:117 arg1 technology 126:135 antibody-lectin sandwich array (ALSA) technology 88:135 antibody-lectin sandwich array (ALSA) technology 88:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 12 80 theme microanalytical 1978:1992 arg1 approach 1994:2001 this microanalytical approach 1973:2001 this microanalytical approach 1973:2001 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 2 81 theme tumor 351:355 arg1 formation 357:365 tumor formation 351:365 tumor formation 351:365 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 0 82 theme ALSA 120:123 arg1 technology 126:135 antibody-lectin sandwich array (ALSA) technology 88:135 antibody-lectin sandwich array (ALSA) technology 88:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 6 83 theme content 1090:1096 arg1 characterization 1061:1076 the characterization 1057:1076 the characterization of N-glycan content of hGGT1 in complex biological samples 1057:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 10 84 theme lectin 1463:1468 arg1 patterns 1478:1485 The lectin binding patterns 1459:1485 The lectin binding patterns obtained with the ALSA platform 1459:1517 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 10 84 theme lectin 1463:1468 arg1 consistent 1523:1532 consistent 1523:1532 consistent 1523:1532 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 7 85 attach isolated 1178:1185 arg2 hGGT1 1165:1169 hGGT1 1165:1169 hGGT1 1165:1169 RESULTS Our data show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform. 25479762 7 85 attach isolated 1178:1185 arg1 detergent 1192:1200 detergent 1192:1200 detergent extracted membrane proteins by binding to the ALSA platform 1192:1260 RESULTS Our data show that hGGT1 can be isolated from detergent extracted membrane proteins by binding to the ALSA platform. 25479762 1 86 theme extracellular 239:251 arg1 glutathione 253:263 extracellular glutathione 239:263 extracellular glutathione 239:263 BACKGROUND γ-Glutamyl transpeptidase 1 (GGT1) is an N-glycosylated membrane protein that catabolizes extracellular glutathione and other γ-glutamyl-containing substrates. 25479762 5 87 theme protein 787:793 arg1 quantities 764:773 the large quantities 754:773 the large quantities of purified protein 754:793 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 11 88 theme many 1943:1946 arg1 tumors 1948:1953 many tumors 1943:1953 many tumors 1943:1953 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 0 89 theme glycosylation 22:34 arg1 patterns 36:43 distinct glycosylation patterns 13:43 distinct glycosylation patterns 13:43 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 12 90 from hGGT1 2181:2185 arg1 changes 2140:2146 disease-related changes 2124:2146 disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens 2124:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 5 91 theme large 758:762 arg1 quantities 764:773 the large quantities 754:773 the large quantities of purified protein 754:793 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 9 92 theme human 1361:1365 arg1 tissue 1373:1378 normal human liver tissue 1354:1378 normal human liver tissue 1354:1378 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 6 93 from samples 1129:1135 arg1 content 1090:1096 N-glycan content 1081:1096 N-glycan content of hGGT1 in complex biological samples 1081:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 2 94 theme cell 411:414 arg1 surface 396:402 the surface 392:402 the surface of the cell 392:414 In a variety of disease states, including tumor formation, the enzyme is shed from the surface of the cell and can be detected in serum. 25479762 0 95 theme γ-glutamyl 54:63 arg1 transpeptidase 65:78 human γ-glutamyl transpeptidase 1 48:80 human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology 48:135 Detection of distinct glycosylation patterns on human γ-glutamyl transpeptidase 1 using antibody-lectin sandwich array (ALSA) technology. 25479762 9 96 theme normal 1381:1386 arg1 tissue 1401:1406 normal human kidney tissue 1381:1406 normal human kidney tissue 1381:1406 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 10 97 theme ALSA 1505:1508 arg1 platform 1510:1517 the ALSA platform 1501:1517 the ALSA platform 1501:1517 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 5 98 dep content 843:849 arg1 poses 851:855 poses 851:855 poses a significant challenge for biomarker development 851:905 However, the large quantities of purified protein required to fully characterize the carbohydrate content poses a significant challenge for biomarker development. 25479762 6 99 from hGGT1 1101:1105 arg1 samples 1129:1135 complex biological samples 1110:1135 complex biological samples 1110:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 11 100 theme modification 1888:1899 arg1 efficacy 1798:1805 its efficacy 1794:1805 its efficacy in specifically recognizing high-mannose-type N-glycans 1794:1861 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 11 100 theme modification 1888:1899 arg1 class 1866:1870 a class 1864:1870 a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors 1864:1953 We also validate the implementation of the Microcystis aeruginosa lectin, microvirin, in this platform and provide refined evidence for its efficacy in specifically recognizing high-mannose-type N-glycans, a class of carbohydrate modification that is distinctive of hGGT1 expressed by many tumors. 25479762 9 101 theme kidney 1394:1399 arg1 tissue 1401:1406 normal human kidney tissue 1381:1406 normal human kidney tissue 1381:1406 We probed hGGT1 from normal human liver tissue, normal human kidney tissue, and hGGT1 expressed in the yeast Pichia pastoris. 25479762 6 102 theme complex 1110:1116 arg1 samples 1129:1135 complex biological samples 1110:1135 complex biological samples 1110:1135 Herein, we investigated a new antibody-lectin sandwich array (ALSA) platform to determine whether this microanalytical technique could be applied to the characterization of N-glycan content of hGGT1 in complex biological samples. 25479762 10 103 theme hGGT1 1543:1547 arg1 composition 1558:1568 the hGGT1 N-glycan composition 1539:1568 the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources 1539:1655 The lectin binding patterns obtained with the ALSA platform are consistent with the hGGT1 N-glycan composition obtained from previous large-scale hGGT1 N-glycan characterizations from these sources. 25479762 12 104 theme treatment 2246:2254 arg1 regimens 2256:2263 targeted treatment regimens 2237:2263 targeted treatment regimens 2237:2263 CONCLUSION Using this microanalytical approach, we provide proof-of-concept for the implementation of ALSA in conducting high-throughput studies aimed at investigating disease-related changes in the glycosylation patterns on hGGT1 with the goal of enhancing clinical diagnoses and targeted treatment regimens. 25479762 4 105 from changes 554:560 arg1 glycans 569:575 the glycans 565:575 the glycans 565:575 Tumor-specific changes in the glycans have also been observed, suggesting that the N-glycans on hGGT1 would be an important biomarker for detecting tumors and monitoring their progression during treatment. 29145098 6 0 theme SGP 1447:1449 arg1 % 1441:1441 % 1441:1441 % 1441:1441 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 0 theme SGP 1447:1449 arg1 % 1488:1488 97.34% 1483:1488 97.34% 1483:1488 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 0 theme SGP 1447:1449 arg1 recoveries 1429:1438 the mean recoveries 1420:1438 the mean recoveries (%) of SGP spiked in the human plasma 1420:1476 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 1 1 theme simple 271:276 arg1 method 278:283 a highly sensitive and simple method 248:283 a highly sensitive and simple method for the quantitative analysis of glycan 248:323 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 7 2 from B 1650:1650 arg1 glycans 1624:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans 1595:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1595:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 2 from B 1650:1650 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 0 3 theme mutant 208:213 arg1 Endo-M-N175Q 222:233 mutant enzyme Endo-M-N175Q 208:233 mutant enzyme Endo-M-N175Q 208:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 4 theme disease 456:462 arg1 biomarkers 471:480 disease glycan biomarkers 456:480 disease glycan biomarkers 456:480 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 0 5 theme Endo-M-N175Q 222:233 arg1 reaction 196:203 transglycosylation reaction 177:203 transglycosylation reaction of mutant enzyme Endo-M-N175Q 177:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 6 theme biomarkers 471:480 arg1 screening 443:451 screening 443:451 screening of disease glycan biomarkers 443:480 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 6 theme biomarkers 471:480 arg1 exploration 341:351 the exploration 337:351 the exploration of protein glycosylation patterns (glycomics) 337:397 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 6 theme biomarkers 471:480 arg1 evaluation 400:409 evaluation 400:409 evaluation of antibody drug stability 400:436 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 5 7 dep d0/d8 1303:1307 arg1 d8/d0 1321:1325 d8/d0 1321:1325 d8/d0 1321:1325 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 5 8 from ratios 1225:1230 arg1 method 1271:1276 the QMERSIL method 1259:1276 the QMERSIL method 1259:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 3 9 theme enzyme 589:594 arg1 QMERSIL 630:636 QMERSIL 630:636 QMERSIL 630:636 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 9 theme enzyme 589:594 arg1 labeling 620:627 mutant enzyme reaction stable isotope labeling 582:627 mutant enzyme reaction stable isotope labeling (QMERSIL) 582:637 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 7 10 theme d0 1587:1588 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 5 11 dep linearity 1292:1300 arg1 R2=0.9999 1310:1318 R2=0.9999 1310:1318 R2=0.9999 1310:1318 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 5 11 dep linearity 1292:1300 arg1 d0/d8 1303:1307 d0/d8 1303:1307 d0/d8 1303:1307 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 5 11 dep linearity 1292:1300 arg1 R2=0.9978 1328:1336 R2=0.9978 1328:1336 R2=0.9978 1328:1336 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 7 12 from ovalbumin 1672:1680 arg1 glycans 1624:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans 1595:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1595:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 12 from ovalbumin 1672:1680 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 8 13 dep labeled 1766:1772 arg1 MPDPZ-Boc-Asn-GlcNAc 1745:1764 MPDPZ-Boc-Asn-GlcNAc 1745:1764 MPDPZ-Boc-Asn-GlcNAc 1745:1764 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 2 14 theme glycomics 546:554 arg1 strategy 556:563 a novel and simplified quantitative glycomics strategy 510:563 a novel and simplified quantitative glycomics strategy 510:563 In this study, we describe a novel and simplified quantitative glycomics strategy. 29145098 5 15 theme peaks 1248:1252 arg1 ratios 1225:1230 The area ratios 1216:1230 The area ratios of the N-glycan peaks from the QMERSIL method 1216:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 7 16 dep ratios 1561:1566 arg1 1:5 1574:1576 1:5 1574:1576 1:5 1574:1576 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 16 dep ratios 1561:1566 arg1 1:1 1569:1571 1:1 1569:1571 1:1 1569:1571 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 16 dep ratios 1561:1566 arg1 5:1 1579:1581 5:1 1579:1581 5:1 1579:1581 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 4 17 dep d0/d8=0.02:20 1175:1187 arg1 to 1172:1173 to 1172:1173 to 1172:1173 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 0 18 theme isotopic 156:163 arg1 labeling 165:172 the stable isotopic labeling 145:172 the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q 145:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 19 theme drug 423:426 arg1 stability 428:436 antibody drug stability 414:436 antibody drug stability 414:436 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 3 20 theme stable 605:610 arg1 QMERSIL 630:636 QMERSIL 630:636 QMERSIL 630:636 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 20 theme stable 605:610 arg1 labeling 620:627 mutant enzyme reaction stable isotope labeling 582:627 mutant enzyme reaction stable isotope labeling (QMERSIL) 582:637 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 8 21 theme theoretical 1817:1827 arg1 values 1829:1834 the theoretical values 1813:1834 the theoretical values (1:1, 1:5, 5:1) 1813:1850 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 5 22 from method 1271:1276 arg1 ratios 1225:1230 The area ratios 1216:1230 The area ratios of the N-glycan peaks from the QMERSIL method 1216:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 5 22 from method 1271:1276 arg1 peaks 1248:1252 the N-glycan peaks 1235:1252 the N-glycan peaks from the QMERSIL method 1235:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 8 23 theme intensity 1710:1718 arg1 equal 1796:1800 equal 1796:1800 equal 1796:1800 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 8 23 theme intensity 1710:1718 arg1 ratios 1687:1692 The ratios 1683:1692 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans 1683:1782 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 3 24 theme structure 879:887 arg1 N-glycans 652:660 the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure 648:887 the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure 648:887 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 9 25 theme relative 1891:1898 arg1 comparison 1913:1922 the relative quantitative comparison 1887:1922 the relative quantitative comparison of the N-Linked oligosaccharides in human plasma 1887:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 6 26 theme 6.12 1409:1412 arg1 % 1413:1413 6.12% 1409:1413 6.12% 1409:1413 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 7 27 theme bovine 1653:1658 arg1 fetuin 1660:1665 bovine fetuin 1653:1665 bovine fetuin 1653:1665 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 28 theme molar 1555:1559 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 4 29 theme complex 1052:1058 arg1 type 1060:1063 the complex type 1048:1063 the complex type 1048:1063 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 3 30 theme positive 863:870 arg1 structure 879:887 a positive charge structure 861:887 a positive charge structure 861:887 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 0 31 theme Quantitative 66:77 arg1 glycomics 79:87 Quantitative glycomics 66:87 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 0:234 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 32 theme protein 356:362 arg1 glycomics 388:396 glycomics 388:396 glycomics 388:396 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 32 theme protein 356:362 arg1 patterns 378:385 protein glycosylation patterns 356:385 protein glycosylation patterns (glycomics) 356:397 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 2 33 theme simplified 522:531 arg1 strategy 556:563 a novel and simplified quantitative glycomics strategy 510:563 a novel and simplified quantitative glycomics strategy 510:563 In this study, we describe a novel and simplified quantitative glycomics strategy. 29145098 1 34 theme patterns 378:385 arg1 screening 443:451 screening 443:451 screening of disease glycan biomarkers 443:480 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 34 theme patterns 378:385 arg1 exploration 341:351 the exploration 337:351 the exploration of protein glycosylation patterns (glycomics) 337:397 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 34 theme patterns 378:385 arg1 evaluation 400:409 evaluation 400:409 evaluation of antibody drug stability 400:436 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 3 35 theme mass 963:966 arg1 spectrometry 968:979 mass spectrometry 963:979 mass spectrometry 963:979 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 2 36 theme novel 512:516 arg1 strategy 556:563 a novel and simplified quantitative glycomics strategy 510:563 a novel and simplified quantitative glycomics strategy 510:563 In this study, we describe a novel and simplified quantitative glycomics strategy. 29145098 9 37 theme oligosaccharides 1940:1955 arg1 comparison 1913:1922 the relative quantitative comparison 1887:1922 the relative quantitative comparison of the N-Linked oligosaccharides in human plasma 1887:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 0 38 theme ionization 108:117 arg1 spectrometry 124:135 electrospray ionization mass spectrometry 95:135 electrospray ionization mass spectrometry 95:135 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 4 39 dep range 1162:1166 arg1 d0/d8=0.02:20 1175:1187 d0/d8=0.02:20 1175:1187 d0/d8=0.02:20 1175:1187 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 7 40 theme MPDPZ-Boc-Asn-GlcNAc-labeled 1595:1622 arg1 glycans 1624:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans 1595:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1595:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 9 41 from comparison 1913:1922 arg1 plasma 1966:1971 human plasma 1960:1971 human plasma 1960:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 9 42 theme human 1960:1964 arg1 plasma 1966:1971 human plasma 1960:1971 human plasma 1960:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 5 43 theme area 1220:1223 arg1 ratios 1225:1230 The area ratios 1216:1230 The area ratios of the N-glycan peaks from the QMERSIL method 1216:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 4 44 gly sialylglycopeptide 1020:1037 arg1 type 1060:1063 the complex type 1048:1063 the complex type 1048:1063 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 4 44 gly sialylglycopeptide 1020:1037 arg2 SGP 1040:1042 SGP 1040:1042 SGP 1040:1042 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 4 44 gly sialylglycopeptide 1020:1037 arg2 sialylglycopeptide 1020:1037 The sialylglycopeptide 1016:1037 The sialylglycopeptide (SGP) of the complex type 1016:1063 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 3 45 theme with 662:665 arg1 N-glycans 652:660 the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure 648:887 the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure 648:887 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 1 46 theme glycan 318:323 arg1 analysis 306:313 the quantitative analysis 289:313 the quantitative analysis of glycan 289:323 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 0 47 theme N-glycan 56:63 arg1 strategy 20:27 A novel, simplified strategy 0:27 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 0:234 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 0 48 theme relative 32:39 arg1 quantification 41:54 relative quantification 32:54 relative quantification N-glycan 32:63 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 0 49 dep strategy 20:27 arg1 glycomics 79:87 Quantitative glycomics 66:87 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 0:234 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 4 50 theme linear 1147:1152 arg1 range 1162:1166 a linear dynamic range 1145:1166 a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude 1145:1213 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 5 51 theme QMERSIL 1263:1269 arg1 method 1271:1276 the QMERSIL method 1259:1276 the QMERSIL method 1259:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 3 52 dep N-glycans 652:660 arg1 d0- 691:693 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 d0- 691:693 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 52 dep N-glycans 652:660 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 9 53 from oligosaccharides 1940:1955 arg1 plasma 1966:1971 human plasma 1960:1971 human plasma 1960:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 4 54 theme relative 1118:1125 arg1 quantitation 1127:1138 the relative quantitation 1114:1138 the relative quantitation 1114:1138 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 3 55 theme deuterated 699:708 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 5 56 theme N-glycan 1239:1246 arg1 peaks 1248:1252 the N-glycan peaks 1235:1252 the N-glycan peaks from the QMERSIL method 1235:1276 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 4 57 used used 1069:1072 arg2 SGP 1040:1042 SGP 1040:1042 SGP 1040:1042 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 4 57 used used 1069:1072 arg2 sialylglycopeptide 1020:1037 The sialylglycopeptide 1016:1037 The sialylglycopeptide (SGP) of the complex type 1016:1063 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 0 58 theme enzyme 215:220 arg1 Endo-M-N175Q 222:233 mutant enzyme Endo-M-N175Q 208:233 mutant enzyme Endo-M-N175Q 208:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 59 theme method 278:283 arg1 lack 240:243 The lack 236:243 The lack of a highly sensitive and simple method for the quantitative analysis of glycan 236:323 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 60 theme glycan 464:469 arg1 biomarkers 471:480 disease glycan biomarkers 456:480 disease glycan biomarkers 456:480 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 7 61 from fetuin 1660:1665 arg1 glycans 1624:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans 1595:1630 MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1595:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 61 from fetuin 1660:1665 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 4 62 theme orders 1195:1200 arg1 range 1162:1166 a linear dynamic range 1145:1166 a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude 1145:1213 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 3 63 theme d8- 711:713 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 64 theme mutant 582:587 arg1 QMERSIL 630:636 QMERSIL 630:636 QMERSIL 630:636 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 64 theme mutant 582:587 arg1 labeling 620:627 mutant enzyme reaction stable isotope labeling 582:627 mutant enzyme reaction stable isotope labeling (QMERSIL) 582:637 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 65 theme -1,1-dimethyl-piperazin-1-ium 755:783 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 6 66 theme accuracy 1364:1371 arg1 precisions 1379:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 66 theme accuracy 1364:1371 arg1 less 1399:1402 less 1399:1402 less 1399:1402 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 3 67 theme 2,4-Dinitro-5-piperazin-1-yl-phenyl 719:753 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 0 68 theme stable 149:154 arg1 labeling 165:172 the stable isotopic labeling 145:172 the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q 145:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 3 69 theme isotope 612:618 arg1 QMERSIL 630:636 QMERSIL 630:636 QMERSIL 630:636 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 69 theme isotope 612:618 arg1 labeling 620:627 mutant enzyme reaction stable isotope labeling 582:627 mutant enzyme reaction stable isotope labeling (QMERSIL) 582:637 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 2 70 theme quantitative 533:544 arg1 strategy 556:563 a novel and simplified quantitative glycomics strategy 510:563 a novel and simplified quantitative glycomics strategy 510:563 In this study, we describe a novel and simplified quantitative glycomics strategy. 29145098 3 71 theme -Boc-asparaginyl-N-acetyl-d-glucosamine 792:830 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 1 72 theme antibody 414:421 arg1 stability 428:436 antibody drug stability 414:436 antibody drug stability 414:436 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 3 73 theme reaction 596:603 arg1 QMERSIL 630:636 QMERSIL 630:636 QMERSIL 630:636 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 73 theme reaction 596:603 arg1 labeling 620:627 mutant enzyme reaction stable isotope labeling 582:627 mutant enzyme reaction stable isotope labeling (QMERSIL) 582:637 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 74 theme MPDPZ 786:790 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 0 75 theme transglycosylation 177:194 arg1 reaction 196:203 transglycosylation reaction 177:203 transglycosylation reaction of mutant enzyme Endo-M-N175Q 177:233 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 76 theme stability 428:436 arg1 screening 443:451 screening 443:451 screening of disease glycan biomarkers 443:480 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 76 theme stability 428:436 arg1 exploration 341:351 the exploration 337:351 the exploration of protein glycosylation patterns (glycomics) 337:397 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 76 theme stability 428:436 arg1 evaluation 400:409 evaluation 400:409 evaluation of antibody drug stability 400:436 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 3 77 theme glycosynthase 901:913 arg1 reaction 949:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 9 78 used used 1878:1881 arg2 method 1867:1872 this method 1862:1872 this method 1862:1872 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 4 79 theme magnitude 1205:1213 arg1 orders 1195:1200 3 orders 1193:1200 3 orders of magnitude 1193:1213 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 8 80 theme relative 1701:1708 arg1 intensity 1710:1718 the relative intensity 1697:1718 the relative intensity 1697:1718 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 3 81 dep Quantitation 566:577 arg1 label 642:646 label 642:646 to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry 639:979 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 7 82 theme various 1547:1553 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 9 83 theme quantitative 1900:1911 arg1 comparison 1913:1922 the relative quantitative comparison 1887:1922 the relative quantitative comparison of the N-Linked oligosaccharides in human plasma 1887:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 5 84 theme good 1287:1290 arg1 linearity 1292:1300 a good linearity 1285:1300 a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978) 1285:1337 The area ratios of the N-glycan peaks from the QMERSIL method showed a good linearity (d0/d8, R2=0.9999; d8/d0, R2=0.9978). 29145098 3 85 theme Boc-Asn-GlcNAc 833:846 arg1 acceptor 849:856 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 acceptor 849:856 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 3 86 theme transglycosylation 930:947 arg1 reaction 949:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 8 87 dep values 1829:1834 arg1 5:1 1847:1849 5:1 1847:1849 5:1 1847:1849 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 8 87 dep values 1829:1834 arg1 1:5 1842:1844 1:5 1842:1844 1:5 1842:1844 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 8 87 dep values 1829:1834 arg1 1:1 1837:1839 1:1 1837:1839 1:1 1837:1839 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 3 88 theme charge 872:877 arg1 structure 879:887 a positive charge structure 861:887 a positive charge structure 861:887 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 0 89 theme electrospray 95:106 arg1 spectrometry 124:135 electrospray ionization mass spectrometry 95:135 electrospray ionization mass spectrometry 95:135 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 1 90 theme glycosylation 364:376 arg1 glycomics 388:396 glycomics 388:396 glycomics 388:396 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 1 90 theme glycosylation 364:376 arg1 patterns 378:385 protein glycosylation patterns 356:385 protein glycosylation patterns (glycomics) 356:397 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 3 91 theme Endo-M-N175Q 916:927 arg1 reaction 949:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 the glycosynthase (Endo-M-N175Q) transglycosylation reaction 897:956 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 9 92 theme N-Linked 1931:1938 arg1 oligosaccharides 1940:1955 the N-Linked oligosaccharides 1927:1955 the N-Linked oligosaccharides in human plasma 1927:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 0 93 theme mass 119:122 arg1 spectrometry 124:135 electrospray ionization mass spectrometry 95:135 electrospray ionization mass spectrometry 95:135 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 8 94 theme labeled 1766:1772 arg1 N-glycans 1774:1782 the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans 1728:1782 the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans 1728:1782 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 7 95 theme ribonuclease 1637:1648 arg1 B 1650:1650 ribonuclease B 1637:1650 ribonuclease B 1637:1650 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 8 96 dep close 1804:1808 arg1 values 1829:1834 the theoretical values 1813:1834 the theoretical values (1:1, 1:5, 5:1) 1813:1850 The ratios of the relative intensity between the isotopically MPDPZ-Boc-Asn-GlcNAc labeled N-glycans were almost equal a close to the theoretical values (1:1, 1:5, 5:1). 29145098 0 97 theme novel 2:6 arg1 strategy 20:27 A novel, simplified strategy 0:27 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 0:234 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 7 98 theme glycans 1624:1630 arg1 ratios 1561:1566 various molar ratios 1547:1566 various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin 1547:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 6 99 theme reproducibility 1344:1358 arg1 precisions 1379:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 99 theme reproducibility 1344:1358 arg1 less 1399:1402 less 1399:1402 less 1399:1402 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 1 100 theme quantitative 293:304 arg1 analysis 306:313 the quantitative analysis 289:313 the quantitative analysis of glycan 289:323 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 29145098 6 101 theme mean 1424:1427 arg1 % 1441:1441 % 1441:1441 % 1441:1441 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 101 theme mean 1424:1427 arg1 % 1488:1488 97.34% 1483:1488 97.34% 1483:1488 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 101 theme mean 1424:1427 arg1 recoveries 1429:1438 the mean recoveries 1420:1438 the mean recoveries (%) of SGP spiked in the human plasma 1420:1476 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 3 102 theme comparative 993:1003 arg1 glycomics 1005:1013 comparative glycomics 993:1013 comparative glycomics 993:1013 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 4 103 theme type 1060:1063 arg1 SGP 1040:1042 SGP 1040:1042 SGP 1040:1042 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 4 103 theme type 1060:1063 arg1 sialylglycopeptide 1020:1037 The sialylglycopeptide 1016:1037 The sialylglycopeptide (SGP) of the complex type 1016:1063 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 6 104 theme assay 1373:1377 arg1 precisions 1379:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions 1340:1388 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 6 104 theme assay 1373:1377 arg1 less 1399:1402 less 1399:1402 less 1399:1402 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 0 105 theme quantification 41:54 arg1 N-glycan 56:63 relative quantification N-glycan 32:63 relative quantification N-glycan 32:63 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 9 106 from plasma 1966:1971 arg1 comparison 1913:1922 the relative quantitative comparison 1887:1922 the relative quantitative comparison of the N-Linked oligosaccharides in human plasma 1887:1971 Finally, this method was used for the relative quantitative comparison of the N-Linked oligosaccharides in human plasma. 29145098 3 107 theme nondeuterated 676:688 arg1 d0- 691:693 either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor 667:856 d0- 691:693 Quantitation by mutant enzyme reaction stable isotope labeling (QMERSIL) to label the N-glycans with either a nondeuterated (d0-) or deuterated (d8-) 4-(2,4-Dinitro-5-piperazin-1-yl-phenyl)-1,1-dimethyl-piperazin-1-ium (MPDPZ)-Boc-asparaginyl-N-acetyl-d-glucosamine (Boc-Asn-GlcNAc) acceptor of a positive charge structure through the glycosynthase (Endo-M-N175Q) transglycosylation reaction with mass spectrometry facilitates comparative glycomics. 29145098 7 108 from ratios 1561:1566 arg1 B 1650:1650 ribonuclease B 1637:1650 ribonuclease B 1637:1650 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 108 from ratios 1561:1566 arg1 fetuin 1660:1665 bovine fetuin 1653:1665 bovine fetuin 1653:1665 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 7 108 from ratios 1561:1566 arg1 ovalbumin 1672:1680 ovalbumin 1672:1680 ovalbumin 1672:1680 Moreover, the QMERSIL using LC-MS/MS was evaluated with various molar ratios (1:1, 1:5, 5:1) of d0(d8)- MPDPZ-Boc-Asn-GlcNAc-labeled glycans from ribonuclease B, bovine fetuin, and ovalbumin. 29145098 6 109 theme human 1465:1469 arg1 plasma 1471:1476 the human plasma 1461:1476 the human plasma 1461:1476 The reproducibility and accuracy assay precisions were all less than 6.12%, and the mean recoveries (%) of SGP spiked in the human plasma were 97.34%. 29145098 0 110 dep novel 2:6 arg1 simplified 9:18 simplified 9:18 simplified 9:18 A novel, simplified strategy of relative quantification N-glycan: Quantitative glycomics using electrospray ionization mass spectrometry through the stable isotopic labeling by transglycosylation reaction of mutant enzyme Endo-M-N175Q. 29145098 4 111 theme dynamic 1154:1160 arg1 range 1162:1166 a linear dynamic range 1145:1166 a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude 1145:1213 The sialylglycopeptide (SGP) of the complex type was used to demonstrate that QMERSIL facilitates the relative quantitation over a linear dynamic range (up to d0/d8=0.02:20) of 3 orders of magnitude. 29145098 1 112 theme sensitive 257:265 arg1 method 278:283 a highly sensitive and simple method 248:283 a highly sensitive and simple method for the quantitative analysis of glycan 248:323 The lack of a highly sensitive and simple method for the quantitative analysis of glycan has impeded the exploration of protein glycosylation patterns (glycomics), evaluation of antibody drug stability, and screening of disease glycan biomarkers. 27387015 0 0 theme Boletus 93:99 arg1 snicus 101:106 Boletus snicus 93:106 Boletus snicus 93:106 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 8 1 from effects 1013:1019 arg1 glycation 1024:1032 glycation 1024:1032 glycation 1024:1032 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 7 2 theme antiglycation 891:903 arg1 activities 905:914 The antiglycation activities 887:914 The antiglycation activities of BSP-1b and BSP-2b 887:935 The antiglycation activities of BSP-1b and BSP-2b were investigated. 27387015 5 3 theme BSP-2b 585:590 arg1 compositions 569:580 The monosaccharide compositions 550:580 The monosaccharide compositions of BSP-2b 550:590 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 6 4 contain had 854:856 arg1 BSP-2b 847:852 BSP-2b 847:852 BSP-2b 847:852 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 6 4 contain had 854:856 arg2 content 863:869 high content 858:869 high content of uronic acid 858:884 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 5 5 theme 10.70:6.95:12.05:12.57:1.83:1.00 707:738 arg1 ratio 698:702 the molar ratio 688:702 the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00 688:738 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 4 6 theme small 484:488 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 6 theme small 484:488 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 6 theme small 484:488 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 6 theme small 484:488 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 6 theme small 484:488 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 6 7 theme pyranose 819:826 arg1 structure 833:841 pyranose ring structure 819:841 pyranose ring structure 819:841 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 2 8 theme BS 269:270 arg1 body 261:264 the body 257:264 the body of BS 257:270 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 9 9 theme stronger 1088:1095 arg1 activity 1111:1118 stronger antiglycation activity 1088:1118 stronger antiglycation activity 1088:1118 BSP-2b showed stronger antiglycation activity than BSP-1b. 27387015 3 10 theme molecular 347:355 arg1 weight 357:362 molecular weight 347:362 molecular weight of BSP-1b and BSP-2b 347:383 The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. 27387015 6 11 theme NMR 762:764 arg1 analysis 766:773 NMR analysis 762:773 NMR analysis 762:773 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 8 12 theme high 997:1000 arg1 effects 1013:1019 high inhibitory effects 997:1019 high inhibitory effects on glycation 997:1032 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 9 13 theme antiglycation 1097:1109 arg1 activity 1111:1118 stronger antiglycation activity 1088:1118 stronger antiglycation activity 1088:1118 BSP-2b showed stronger antiglycation activity than BSP-1b. 27387015 8 14 theme inhibitory 1002:1011 arg1 effects 1013:1019 high inhibitory effects 997:1019 high inhibitory effects on glycation 997:1032 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 4 15 theme large 454:458 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 5 16 theme glucuronic 609:618 arg1 mannose 600:606 mannose 600:606 mannose 600:606 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 16 theme glucuronic 609:618 arg1 acid 620:623 glucuronic acid 609:623 glucuronic acid 609:623 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 4 17 with heteropolysaccharide 426:445 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 17 with heteropolysaccharide 426:445 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 17 with heteropolysaccharide 426:445 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 17 with heteropolysaccharide 426:445 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 17 with heteropolysaccharide 426:445 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 17 with heteropolysaccharide 426:445 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 1 18 theme important 156:164 arg1 species 175:181 the commercially important mushroom species 139:181 the commercially important mushroom species 139:181 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 0 19 from activities 48:57 arg1 snicus 101:106 Boletus snicus 93:106 Boletus snicus 93:106 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 6 20 theme high 858:861 arg1 content 863:869 high content 858:869 high content of uronic acid 858:884 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 1 21 theme mushroom 166:173 arg1 species 175:181 the commercially important mushroom species 139:181 the commercially important mushroom species 139:181 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 3 22 theme BSP-1b 367:372 arg1 weight 357:362 molecular weight 347:362 molecular weight of BSP-1b and BSP-2b 347:383 The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. 27387015 6 23 theme acid 881:884 arg1 content 863:869 high content 858:869 high content of uronic acid 858:884 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 8 24 theme dose-dependent 1048:1061 arg1 responses 1063:1071 dose-dependent responses 1048:1071 dose-dependent responses 1048:1071 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 5 25 theme monosaccharide 554:567 arg1 compositions 569:580 The monosaccharide compositions 550:580 The monosaccharide compositions of BSP-2b 550:590 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 2 26 theme Sephadex 294:301 arg1 chromatography 316:329 DEAE-cellulose and Sephadex G-100 column chromatography 275:329 chromatography 316:329 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 1 27 theme species 175:181 arg1 species 175:181 the commercially important mushroom species 139:181 the commercially important mushroom species 139:181 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 1 27 theme species 175:181 arg1 one 132:134 one 132:134 one 132:134 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 0 28 theme Preparation 0:10 arg1 activities 48:57 Preparation, characterization and antiglycation activities 0:57 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 0:107 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 5 29 theme glucosamine 626:636 arg1 mannose 600:606 mannose 600:606 mannose 600:606 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 29 theme glucosamine 626:636 arg1 hydrochloride 638:650 glucosamine hydrochloride 626:650 glucosamine hydrochloride 626:650 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 7 30 theme BSP-2b 930:935 arg1 activities 905:914 The antiglycation activities 887:914 The antiglycation activities of BSP-1b and BSP-2b 887:935 The antiglycation activities of BSP-1b and BSP-2b were investigated. 27387015 3 31 theme weight 357:362 arg1 average 336:342 The average 332:342 The average of molecular weight of BSP-1b and BSP-2b 332:383 The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. 27387015 3 31 theme weight 357:362 arg1 59.21kDa 390:397 59.21kDa 390:397 59.21kDa 390:397 The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. 27387015 4 32 theme hydrochloride 521:533 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 32 theme hydrochloride 521:533 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 32 theme hydrochloride 521:533 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 32 theme hydrochloride 521:533 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 32 theme hydrochloride 521:533 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 32 theme hydrochloride 521:533 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 2 33 theme DEAE-cellulose 275:288 arg1 chromatography 316:329 DEAE-cellulose and Sephadex G-100 column chromatography 275:329 chromatography 316:329 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 6 34 theme ring 828:831 arg1 structure 833:841 pyranose ring structure 819:841 pyranose ring structure 819:841 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 0 35 theme characterization 13:28 arg1 activities 48:57 Preparation, characterization and antiglycation activities 0:57 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 0:107 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 5 36 theme molar 692:696 arg1 ratio 698:702 the molar ratio 688:702 the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00 688:738 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 6 37 theme uronic 874:879 arg1 acid 881:884 uronic acid 874:884 uronic acid 874:884 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 4 38 theme arabinose 539:547 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 38 theme arabinose 539:547 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 38 theme arabinose 539:547 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 38 theme arabinose 539:547 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 38 theme arabinose 539:547 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 38 theme arabinose 539:547 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 0 39 theme antiglycation 34:46 arg1 activities 48:57 Preparation, characterization and antiglycation activities 0:57 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 0:107 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 4 40 theme mannose 500:506 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 40 theme mannose 500:506 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 40 theme mannose 500:506 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 40 theme mannose 500:506 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 40 theme mannose 500:506 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 40 theme mannose 500:506 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 3 41 theme BSP-2b 378:383 arg1 weight 357:362 molecular weight 347:362 molecular weight of BSP-1b and BSP-2b 347:383 The average of molecular weight of BSP-1b and BSP-2b were 59.21kDa and 128.74kDa. 27387015 5 42 with mannose 600:606 arg1 ratio 698:702 the molar ratio 688:702 the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00 688:738 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg1 compositions 569:580 The monosaccharide compositions 550:580 The monosaccharide compositions of BSP-2b 550:590 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 galactose 662:670 galactose 662:670 galactose 662:670 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 mannose 600:606 mannose 600:606 mannose 600:606 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 hydrochloride 638:650 glucosamine hydrochloride 626:650 glucosamine hydrochloride 626:650 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 arabinose 673:681 arabinose 673:681 arabinose 673:681 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 glucose 653:659 glucose 653:659 glucose 653:659 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 5 43 contain contain 592:598 arg2 acid 620:623 glucuronic acid 609:623 glucuronic acid 609:623 The monosaccharide compositions of BSP-2b contain mannose, glucuronic acid, glucosamine hydrochloride, glucose, galactose, arabinose with the molar ratio of 10.70:6.95:12.05:12.57:1.83:1.00. 27387015 4 44 theme glucose 470:476 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 44 theme glucose 470:476 arg1 amount 490:495 a small amount 482:495 a small amount of mannose, glucosamine hydrochloride and arabinose 482:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 44 theme glucose 470:476 arg1 arabinose 539:547 arabinose 539:547 arabinose 539:547 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 44 theme glucose 470:476 arg1 number 460:465 a large number 452:465 a large number of glucose 452:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 44 theme glucose 470:476 arg1 glucose 470:476 glucose 470:476 glucose 470:476 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 44 theme glucose 470:476 arg1 mannose 500:506 mannose 500:506 mannose 500:506 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 4 45 theme glucosamine 509:519 arg1 hydrochloride 521:533 glucosamine hydrochloride 509:533 glucosamine hydrochloride 509:533 BSP-1b is a heteropolysaccharide with a large number of glucose and a small amount of mannose, glucosamine hydrochloride and arabinose. 27387015 7 46 theme BSP-1b 919:924 arg1 activities 905:914 The antiglycation activities 887:914 The antiglycation activities of BSP-1b and BSP-2b 887:935 The antiglycation activities of BSP-1b and BSP-2b were investigated. 27387015 2 47 theme column 309:314 arg1 chromatography 316:329 DEAE-cellulose and Sephadex G-100 column chromatography 275:329 chromatography 316:329 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 1 48 theme Boletus 109:115 arg1 BS 125:126 BS 125:126 BS 125:126 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 1 48 theme Boletus 109:115 arg1 snicus 117:122 Boletus snicus 109:122 Boletus snicus (BS) 109:127 Boletus snicus (BS) is one of the commercially important mushroom species. 27387015 2 49 dep polysaccharides 188:202 arg1 polysaccharides 188:202 Two polysaccharides 184:202 Two polysaccharides (BSP-1b and BSP-2b) 184:222 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 2 49 dep polysaccharides 188:202 arg1 BSP-2b 216:221 BSP-2b 216:221 BSP-2b 216:221 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 2 49 dep polysaccharides 188:202 arg1 BSP-1b 205:210 BSP-1b 205:210 BSP-1b 205:210 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 6 50 theme FTIR 745:748 arg1 spectra 750:756 The FTIR spectra 741:756 The FTIR spectra 741:756 The FTIR spectra and NMR analysis demonstrated that BSP-1b and BSP-2b existed pyranose ring structure and BSP-2b had high content of uronic acid. 27387015 2 51 theme G-100 303:307 arg1 chromatography 316:329 DEAE-cellulose and Sephadex G-100 column chromatography 275:329 chromatography 316:329 Two polysaccharides (BSP-1b and BSP-2b) were extracted and purified from the body of BS by DEAE-cellulose and Sephadex G-100 column chromatography. 27387015 10 52 theme glycation 1222:1230 arg1 end-products 1232:1243 advanced glycation end-products 1213:1243 advanced glycation end-products 1213:1243 This study indicated that the BSP-2b could effectively inhibit the formation of advanced glycation end-products. 27387015 0 53 theme polysaccharides 72:86 arg1 activities 48:57 Preparation, characterization and antiglycation activities 0:57 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 0:107 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 0 54 from snicus 101:106 arg1 activities 48:57 Preparation, characterization and antiglycation activities 0:57 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 0:107 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 0 54 from snicus 101:106 arg1 polysaccharides 72:86 the novel polysaccharides 62:86 the novel polysaccharides from Boletus snicus 62:106 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 8 55 dep showed 968:973 arg1 had 993:995 had 993:995 had high inhibitory effects on glycation 993:1032 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 8 55 dep showed 968:973 arg1 exhibited 1038:1046 exhibited 1038:1046 exhibited dose-dependent responses 1038:1071 The results showed BSP-1b and BSP-2b had high inhibitory effects on glycation and exhibited dose-dependent responses. 27387015 10 56 theme advanced 1213:1220 arg1 end-products 1232:1243 advanced glycation end-products 1213:1243 advanced glycation end-products 1213:1243 This study indicated that the BSP-2b could effectively inhibit the formation of advanced glycation end-products. 27387015 0 57 theme novel 66:70 arg1 polysaccharides 72:86 the novel polysaccharides 62:86 the novel polysaccharides from Boletus snicus 62:106 Preparation, characterization and antiglycation activities of the novel polysaccharides from Boletus snicus. 27387015 10 58 theme end-products 1232:1243 arg1 formation 1200:1208 the formation 1196:1208 the formation of advanced glycation end-products 1196:1243 This study indicated that the BSP-2b could effectively inhibit the formation of advanced glycation end-products. 25527317 4 0 dep Bos 817:819 arg1 taurus 821:826 Bos taurus 817:826 Bos taurus 817:826 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 5 1 theme blood 1105:1109 arg1 vessels 1111:1117 placental blood vessels 1095:1117 placental blood vessels 1095:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 10 2 theme biosynthetic 1602:1613 arg1 pathways 1615:1622 different glycan biosynthetic pathways 1585:1622 different glycan biosynthetic pathways following their split 1585:1644 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 4 3 from embedded 724:731 arg1 resin 736:740 resin 736:740 resin 736:740 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 10 4 contain have 1795:1798 arg1 inter 1759:1763 inter 1759:1763 inter 1759:1763 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 10 4 contain have 1795:1798 arg2 functions 1810:1818 different functions 1800:1818 different functions 1800:1818 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 10 4 contain have 1795:1798 arg1 BNC 1781:1783 placentomal BNC 1769:1783 placentomal BNC 1769:1783 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 1 5 theme glycan 240:245 arg1 structure 247:255 a highly conserved glycan structure 221:255 a highly conserved glycan structure which evolved early in Ruminant phylogenesis 221:300 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 2 6 contain contain 336:342 arg1 Giraffe 303:309 Giraffe 303:309 Giraffe 303:309 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 2 6 contain contain 336:342 arg2 cells 350:354 these cells 344:354 these cells 344:354 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 2 6 contain contain 336:342 arg1 Okapi 315:319 Okapi 315:319 Okapi 315:319 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 3 7 theme cells 491:495 arg1 glycosylation 468:480 the glycosylation 464:480 the glycosylation of these cells 464:495 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 3 8 contain have 553:556 arg2 composition 584:594 a typical ruminant glycan composition 558:594 a typical ruminant glycan composition 558:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 3 8 contain have 553:556 arg1 BNC 543:545 bovine BNC 536:545 bovine BNC which have a typical ruminant glycan composition 536:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 0 9 theme other 96:100 arg1 ruminants 102:110 other ruminants 96:110 other ruminants 96:110 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 5 10 theme villosa 1052:1058 arg1 agglutinins 1060:1070 Dolichos biflorus and Vicia villosa agglutinins 1024:1070 Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 1024:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 4 11 theme bovine 809:814 arg1 Bos 817:819 Bos 817:819 Bos 817:819 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 11 theme bovine 809:814 arg1 placenta 829:836 near-term bovine (Bos taurus) placenta 799:836 near-term bovine (Bos taurus) placenta 799:836 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 3 12 theme bovine 536:541 arg1 BNC 543:545 bovine BNC 536:545 bovine BNC which have a typical ruminant glycan composition 536:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 9 13 from areas 1415:1419 arg1 Bovine 1436:1441 Bovine 1436:1441 Bovine 1436:1441 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 9 13 from areas 1415:1419 arg1 Giraffe 1424:1430 Giraffe 1424:1430 Giraffe 1424:1430 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 5 14 theme αN-acetylgalactosamine 992:1013 arg1 expression 969:978 expression 969:978 expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 969:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 4 15 theme near-term 799:807 arg1 Bos 817:819 Bos 817:819 Bos 817:819 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 15 theme near-term 799:807 arg1 placenta 829:836 near-term bovine (Bos taurus) placenta 799:836 near-term bovine (Bos taurus) placenta 799:836 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 9 16 from cells 1483:1487 arg1 differences 1450:1460 differences 1450:1460 differences from the placentomal cells 1450:1487 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 8 17 dep residues 1388:1395 arg1 were 1375:1378 were 1375:1378 were 1375:1378 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 8 17 dep residues 1388:1395 arg1 as 1372:1373 as 1372:1373 as 1372:1373 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 4 18 theme Giraffa 645:651 arg1 Giraffe 636:642 Giraffe 636:642 Giraffe (Giraffa camelopardalis) 636:667 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 18 theme Giraffa 645:651 arg1 camelopardalis 653:666 Giraffa camelopardalis 645:666 Giraffa camelopardalis 645:666 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 6 19 theme Phytolacca 1194:1203 arg1 agglutinins 1215:1225 Phytolacca americana agglutinins 1194:1225 Phytolacca americana agglutinins 1194:1225 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. 25527317 6 20 theme americana 1205:1213 arg1 agglutinins 1215:1225 Phytolacca americana agglutinins 1194:1225 Phytolacca americana agglutinins 1194:1225 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. 25527317 5 21 theme Giraffe 892:898 arg1 glycans 881:887 the glycans 877:887 the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 877:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 10 22 with Bovidae 1655:1661 arg1 differences 1676:1686 further differences 1668:1686 further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development 1668:1837 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 1 23 theme ruminants 159:167 arg1 placenta 147:154 The placenta 143:154 The placenta of ruminants 143:167 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 10 24 theme different 1585:1593 arg1 pathways 1615:1622 different glycan biosynthetic pathways 1585:1622 different glycan biosynthetic pathways following their split 1585:1644 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 5 25 dep little 956:961 arg1 expression 969:978 expression 969:978 expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 969:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 8 26 theme Most 1304:1307 arg1 glycans 1318:1324 Most N-linked glycans 1304:1324 Most N-linked glycans 1304:1324 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 4 27 theme near 620:623 arg1 term 625:628 near term 620:628 near term 620:628 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 27 theme near 620:623 arg1 placentae 601:609 Two placentae 597:609 Two placentae 597:609 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 0 28 theme binucleate 4:13 arg1 cell 15:18 The binucleate cell 0:18 The binucleate cell of okapi and giraffe placenta 0:48 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 5 29 theme Dolichos 1024:1031 arg1 agglutinins 1060:1070 Dolichos biflorus and Vicia villosa agglutinins 1024:1070 Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 1024:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 8 30 link N-linked 1309:1316 arg1 glycans 1318:1324 Most N-linked glycans 1304:1324 Most N-linked glycans 1304:1324 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 10 31 theme placentomal 1769:1779 arg1 BNC 1781:1783 placentomal BNC 1769:1783 placentomal BNC 1769:1783 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 1 32 theme characteristic 178:191 arg1 BNC 211:213 BNC 211:213 BNC 211:213 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 1 32 theme characteristic 178:191 arg1 cells 204:208 characteristic binucleate cells 178:208 characteristic binucleate cells (BNC) 178:214 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 5 33 theme Vicia 1046:1050 arg1 villosa 1052:1058 Vicia villosa 1046:1058 Vicia villosa 1046:1058 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 6 34 theme Higher 1120:1125 arg1 levels 1127:1132 Higher levels 1120:1132 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins 1120:1225 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. 25527317 4 35 theme lectins 773:779 arg1 panel 761:765 a panel 759:765 a panel of 23 lectins 759:779 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 3 36 gly glycosylation 468:480 arg1 cells 491:495 these cells 485:495 these cells 485:495 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 3 36 gly glycosylation 468:480 arg1 species 506:512 these species 500:512 these species 500:512 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 1 37 theme binucleate 193:202 arg1 BNC 211:213 BNC 211:213 BNC 211:213 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 1 37 theme binucleate 193:202 arg1 cells 204:208 characteristic binucleate cells 178:208 characteristic binucleate cells (BNC) 178:214 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 10 38 theme Giraffidae 1560:1569 arg1 BNC 1571:1573 Giraffidae BNC 1560:1573 Giraffidae BNC 1560:1573 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 5 39 theme placental 1095:1103 arg1 vessels 1111:1117 placental blood vessels 1095:1117 placental blood vessels 1095:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 0 40 theme lectin 115:120 arg1 study 136:140 a lectin histochemical study 113:140 a lectin histochemical study 113:140 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 4 41 from placenta 682:689 arg1 mid 612:614 mid 612:614 mid 612:614 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 41 from placenta 682:689 arg1 term 625:628 near term 620:628 near term 620:628 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 41 from placenta 682:689 arg1 placentae 601:609 Two placentae 597:609 Two placentae 597:609 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 6 42 theme Lycopersicon 1166:1177 arg1 esculentum 1179:1188 Lycopersicon esculentum 1166:1188 Lycopersicon esculentum 1166:1188 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. 25527317 2 43 theme glycan 404:409 arg1 array 411:415 a similar glycan array 394:415 a similar glycan array 394:415 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 3 44 used used 426:429 arg2 We 418:419 We 418:419 We 418:419 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 1 45 theme Ruminant 280:287 arg1 phylogenesis 289:300 Ruminant phylogenesis 280:300 Ruminant phylogenesis 280:300 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 8 46 theme N-linked 1309:1316 arg1 glycans 1318:1324 Most N-linked glycans 1304:1324 Most N-linked glycans 1304:1324 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 0 47 dep shows 50:54 arg1 study 136:140 a lectin histochemical study 113:140 a lectin histochemical study 113:140 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 9 48 theme intercotyledonary 1499:1515 arg1 BNC 1517:1519 no intercotyledonary BNC 1496:1519 no intercotyledonary BNC 1496:1519 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 2 49 theme similar 396:402 arg1 array 411:415 a similar glycan array 394:415 a similar glycan array 394:415 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 10 50 theme further 1668:1674 arg1 differences 1676:1686 further differences 1668:1686 further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development 1668:1837 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 3 51 theme typical 560:566 arg1 composition 584:594 a typical ruminant glycan composition 558:594 a typical ruminant glycan composition 558:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 0 52 theme okapi 23:27 arg1 cell 15:18 The binucleate cell 0:18 The binucleate cell of okapi and giraffe placenta 0:48 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 5 53 located found 868:872 arg2 differences 851:861 Significant differences 839:861 Significant differences 839:861 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 5 53 located found 868:872 arg1 glycans 881:887 the glycans 877:887 the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 877:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 0 54 theme histochemical 122:134 arg1 study 136:140 a lectin histochemical study 113:140 a lectin histochemical study 113:140 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 1 55 contain contains 169:176 arg2 BNC 211:213 BNC 211:213 BNC 211:213 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 1 55 contain contains 169:176 arg1 placenta 147:154 The placenta 143:154 The placenta of ruminants 143:167 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 1 55 contain contains 169:176 arg2 cells 204:208 characteristic binucleate cells 178:208 characteristic binucleate cells (BNC) 178:214 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25527317 3 56 theme ruminant 568:575 arg1 composition 584:594 a typical ruminant glycan composition 558:594 a typical ruminant glycan composition 558:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 3 57 theme lectin 431:436 arg1 histochemistry 438:451 lectin histochemistry 431:451 lectin histochemistry 431:451 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 10 58 theme glycan 1595:1600 arg1 pathways 1615:1622 different glycan biosynthetic pathways 1585:1622 different glycan biosynthetic pathways following their split 1585:1644 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 5 59 dep Dolichos 1024:1031 arg1 biflorus 1033:1040 biflorus 1033:1040 biflorus 1033:1040 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 5 60 theme terminal 983:990 arg1 αN-acetylgalactosamine 992:1013 terminal αN-acetylgalactosamine 983:1013 terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 983:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 2 61 dep Giraffe 303:309 arg1 placentae 321:329 placentae 321:329 placentae 321:329 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 0 62 theme giraffe 33:39 arg1 cell 15:18 The binucleate cell 0:18 The binucleate cell of okapi and giraffe placenta 0:48 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 2 63 contain have 389:392 arg2 array 411:415 a similar glycan array 394:415 a similar glycan array 394:415 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 2 63 contain have 389:392 arg1 they 384:387 they 384:387 they 384:387 Giraffe and Okapi placentae also contain these cells and it is not known whether they have a similar glycan array. 25527317 0 64 dep okapi 23:27 arg1 placenta 41:48 placenta 41:48 placenta 41:48 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 6 65 theme N-acetylglucosamine 1137:1155 arg1 levels 1127:1132 Higher levels 1120:1132 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins 1120:1225 Higher levels of N-acetylglucosamine bound by Lycopersicon esculentum and Phytolacca americana agglutinins were also apparent. 25527317 10 66 theme different 1800:1808 arg1 functions 1810:1818 different functions 1800:1818 different functions 1800:1818 In conclusion, Giraffidae BNC developed different glycan biosynthetic pathways following their split from the Bovidae with further differences evolving as Okapi and Giraffe diverged from each other, affecting both inter and placentomal BNC which may have different functions during development. 25527317 5 67 theme Okapi 904:908 arg1 BNC 910:912 Okapi BNC 904:912 Okapi BNC 904:912 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 4 68 theme term 677:680 arg1 placenta 682:689 two term placenta 673:689 two term placenta 673:689 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 5 69 theme BNC 910:912 arg1 glycans 881:887 the glycans 877:887 the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels 877:1117 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 4 70 theme Okapi 694:698 arg1 mid 612:614 mid 612:614 mid 612:614 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 70 theme Okapi 694:698 arg1 term 625:628 near term 620:628 near term 620:628 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 70 theme Okapi 694:698 arg1 placentae 601:609 Two placentae 597:609 Two placentae 597:609 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 9 71 theme placentomal 1471:1481 arg1 cells 1483:1487 the placentomal cells 1467:1487 the placentomal cells 1467:1487 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 9 72 from Okapi 1538:1542 arg1 apparent 1526:1533 apparent 1526:1533 apparent 1526:1533 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 5 73 theme Significant 839:849 arg1 differences 851:861 Significant differences 839:861 Significant differences 839:861 Significant differences were found in the glycans of Giraffe and Okapi BNC compared with those from the bovine, with little or no expression of terminal αN-acetylgalactosamine bound by Dolichos biflorus and Vicia villosa agglutinins which instead bound to placental blood vessels. 25527317 0 74 theme distinctive 56:66 arg1 glycosylation 68:80 distinctive glycosylation 56:80 distinctive glycosylation 56:80 The binucleate cell of okapi and giraffe placenta shows distinctive glycosylation compared with other ruminants: a lectin histochemical study. 25527317 4 75 from Giraffe 636:642 arg1 mid 612:614 mid 612:614 mid 612:614 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 75 from Giraffe 636:642 arg1 term 625:628 near term 620:628 near term 620:628 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 4 75 from Giraffe 636:642 arg1 placentae 601:609 Two placentae 597:609 Two placentae 597:609 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 3 76 theme glycan 577:582 arg1 composition 584:594 a typical ruminant glycan composition 558:594 a typical ruminant glycan composition 558:594 We have used lectin histochemistry to examine the glycosylation of these cells in these species and compare them with bovine BNC which have a typical ruminant glycan composition. 25527317 4 77 from resin 736:740 arg1 embedded 724:731 embedded 724:731 embedded 724:731 Two placentae, mid and near term, from Giraffe (Giraffa camelopardalis) and two term placenta of Okapi (Okapia johnstoni) were embedded in resin and stained with a panel of 23 lectins and compared with near-term bovine (Bos taurus) placenta. 25527317 9 78 theme Interplacentomal 1398:1413 arg1 areas 1415:1419 Interplacentomal areas 1398:1419 Interplacentomal areas in Giraffe and Bovine 1398:1441 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 8 79 theme fucosyl 1380:1386 arg1 residues 1388:1395 fucosyl residues 1380:1395 fucosyl residues 1380:1395 Most N-linked glycans were similarly expressed in all three species as were fucosyl residues. 25527317 9 80 from apparent 1526:1533 arg1 Okapi 1538:1542 Okapi 1538:1542 Okapi 1538:1542 Interplacentomal areas in Giraffe and Bovine showed differences from the placentomal cells though no intercotyledonary BNC were apparent in Okapi. 25527317 1 81 theme conserved 230:238 arg1 structure 247:255 a highly conserved glycan structure 221:255 a highly conserved glycan structure which evolved early in Ruminant phylogenesis 221:300 The placenta of ruminants contains characteristic binucleate cells (BNC) with a highly conserved glycan structure which evolved early in Ruminant phylogenesis. 25763881 1 0 theme immunological 153:165 arg1 cross-reaction 167:180 immunological cross-reaction 153:180 immunological cross-reaction 153:180 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 0 1 theme genus 73:77 arg1 species 54:60 different potato species 37:60 different potato species of Solanum genus 37:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 0 2 from profiling 10:18 arg1 species 54:60 different potato species 37:60 different potato species of Solanum genus 37:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 7 3 theme therapeutic 1159:1169 arg1 proteins 1171:1178 therapeutic proteins 1159:1178 therapeutic proteins 1159:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 6 4 theme tubers 855:860 arg1 patatins 836:843 patatins 836:843 patatins of potato tubers 836:860 This study found the presence of glycan structures not described previously in patatins of potato tubers, and their glycan profiles significantly differed. 25763881 1 5 theme cross-reaction 167:180 arg1 oligosaccharides 104:119 oligosaccharides 104:119 oligosaccharides 104:119 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 1 5 theme cross-reaction 167:180 arg1 source 143:148 another potential source 125:148 another potential source of immunological cross-reaction between different plant allergens 125:214 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 5 6 theme modified 678:685 arg1 version 687:693 a modified version 676:693 a modified version of on-target derivatization with phenylhydrazine 676:742 For glycan labeling, a modified version of on-target derivatization with phenylhydrazine was applied. 25763881 6 7 theme potato 848:853 arg1 tubers 855:860 potato tubers 848:860 potato tubers 848:860 This study found the presence of glycan structures not described previously in patatins of potato tubers, and their glycan profiles significantly differed. 25763881 4 8 theme laser 608:612 arg1 spectrometry 641:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 0 9 from species 54:60 arg1 patatins 23:30 patatins 23:30 patatins from different potato species of Solanum genus 23:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 0 9 from species 54:60 arg1 profiling 10:18 N-glycome profiling 0:18 N-glycome profiling of patatins from different potato species of Solanum genus 0:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 3 10 gly N-glycosylation 371:385 arg1 proteins 407:414 patatin proteins 399:414 patatin proteins isolated from tubers of different potato species 399:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 7 11 theme correct 990:996 arg1 choice 998:1003 correct choice 990:1003 correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 990:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 4 12 theme matrix-assisted 592:606 arg1 spectrometry 641:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 7 13 theme glycan 1060:1065 arg1 epitopes 1067:1074 specific glycan epitopes 1051:1074 specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 1051:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 7 14 theme potatoes 1127:1134 arg1 utilization 1112:1122 utilization 1112:1122 utilization of potatoes for the manufacture of therapeutic proteins 1112:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 7 14 theme potatoes 1127:1134 arg1 allergy 1089:1095 food allergy 1084:1095 food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 1084:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 3 15 theme proteins 407:414 arg1 profiles 387:394 N-glycosylation profiles 371:394 N-glycosylation profiles of patatin proteins isolated from tubers of different potato species 371:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 5 16 with derivatization 708:721 arg1 phenylhydrazine 728:742 phenylhydrazine 728:742 phenylhydrazine 728:742 For glycan labeling, a modified version of on-target derivatization with phenylhydrazine was applied. 25763881 0 17 theme different 37:45 arg1 species 54:60 different potato species 37:60 different potato species of Solanum genus 37:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 1 18 theme different 190:198 arg1 allergens 206:214 different plant allergens 190:214 different plant allergens 190:214 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 5 19 theme glycan 659:664 arg1 labeling 666:673 glycan labeling 659:673 glycan labeling 659:673 For glycan labeling, a modified version of on-target derivatization with phenylhydrazine was applied. 25763881 2 20 theme Man3 334:337 arg1 Fuc 351:353 Fuc 351:353 Fuc 351:353 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 20 theme Man3 334:337 arg1 GlcNAc2 343:349 composition Man3(Xyl)GlcNAc2 322:349 composition Man3(Xyl)GlcNAc2(Fuc) 322:354 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 21 from glycoprotein 246:257 arg1 potato 262:267 potato 262:267 potato 262:267 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 1 22 theme plant 200:204 arg1 allergens 206:214 different plant allergens 190:214 different plant allergens 190:214 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 0 23 theme N-glycome 0:8 arg1 profiling 10:18 N-glycome profiling 0:18 N-glycome profiling of patatins from different potato species of Solanum genus 0:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 3 24 attach isolated 416:423 arg2 proteins 407:414 patatin proteins 399:414 patatin proteins isolated from tubers of different potato species 399:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 3 24 attach isolated 416:423 arg1 tubers 430:435 tubers 430:435 tubers of different potato species 430:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 2 25 theme composition 322:332 arg1 Fuc 351:353 Fuc 351:353 Fuc 351:353 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 25 theme composition 322:332 arg1 GlcNAc2 343:349 composition Man3(Xyl)GlcNAc2 322:349 composition Man3(Xyl)GlcNAc2(Fuc) 322:354 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 0 26 theme patatins 23:30 arg1 profiling 10:18 N-glycome profiling 0:18 N-glycome profiling of patatins from different potato species of Solanum genus 0:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 4 27 theme PNAGase 556:562 arg1 A 564:564 PNAGase A 556:564 PNAGase A 556:564 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 3 28 theme potato 450:455 arg1 species 457:463 different potato species 440:463 different potato species 440:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 4 29 with digestion 541:549 arg1 A 564:564 PNAGase A 556:564 PNAGase A 556:564 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 2 30 contain have 295:298 arg1 glycoprotein 246:257 the most abundant glycoprotein 228:257 the most abundant glycoprotein in potato 228:267 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 30 contain have 295:298 arg1 Patatin 217:223 Patatin 217:223 Patatin 217:223 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 30 contain have 295:298 arg2 oligosaccharide 303:317 an oligosaccharide 300:317 an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc) 300:354 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 7 31 theme epitopes 1067:1074 arg1 presence 1039:1046 the presence 1035:1046 the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 1035:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 3 32 theme species 457:463 arg1 tubers 430:435 tubers 430:435 tubers of different potato species 430:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 2 33 gly glycoprotein 246:257 arg1 glycoprotein 246:257 the most abundant glycoprotein 228:257 the most abundant glycoprotein in potato 228:267 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 33 gly glycoprotein 246:257 arg1 Patatin 217:223 Patatin 217:223 Patatin 217:223 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 3 34 theme patatin 399:405 arg1 proteins 407:414 patatin proteins 399:414 patatin proteins isolated from tubers of different potato species 399:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 4 35 theme enzymatic 531:539 arg1 digestion 541:549 enzymatic digestion 531:549 enzymatic digestion with PNAGase A 531:564 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 6 36 theme structures 797:806 arg1 presence 778:785 the presence 774:785 the presence of glycan structures not described previously in patatins of potato tubers 774:860 This study found the presence of glycan structures not described previously in patatins of potato tubers, and their glycan profiles significantly differed. 25763881 3 37 theme different 440:448 arg1 species 457:463 different potato species 440:463 different potato species 440:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 3 38 theme N-glycosylation 371:385 arg1 profiles 387:394 N-glycosylation profiles 371:394 N-glycosylation profiles of patatin proteins isolated from tubers of different potato species 371:463 In this work, N-glycosylation profiles of patatin proteins isolated from tubers of different potato species were investigated and compared. 25763881 7 39 theme food 1084:1087 arg1 allergy 1089:1095 food allergy 1084:1095 food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 1084:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 2 40 theme abundant 237:244 arg1 glycoprotein 246:257 the most abundant glycoprotein 228:257 the most abundant glycoprotein in potato 228:267 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 40 theme abundant 237:244 arg1 Patatin 217:223 Patatin 217:223 Patatin 217:223 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 6 41 theme glycan 790:795 arg1 structures 797:806 glycan structures 790:806 glycan structures not described previously in patatins of potato tubers 790:860 This study found the presence of glycan structures not described previously in patatins of potato tubers, and their glycan profiles significantly differed. 25763881 2 42 theme GlcNAc2 343:349 arg1 oligosaccharide 303:317 an oligosaccharide 300:317 an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc) 300:354 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 0 43 theme potato 47:52 arg1 species 54:60 different potato species 37:60 different potato species of Solanum genus 37:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 5 44 theme on-target 698:706 arg1 derivatization 708:721 on-target derivatization 698:721 on-target derivatization with phenylhydrazine 698:742 For glycan labeling, a modified version of on-target derivatization with phenylhydrazine was applied. 25763881 6 45 theme glycan 873:878 arg1 profiles 880:887 their glycan profiles 867:887 their glycan profiles 867:887 This study found the presence of glycan structures not described previously in patatins of potato tubers, and their glycan profiles significantly differed. 25763881 7 46 theme potato 955:960 arg1 patatins 962:969 potato patatins 955:969 potato patatins 955:969 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 5 47 theme derivatization 708:721 arg1 version 687:693 a modified version 676:693 a modified version of on-target derivatization with phenylhydrazine 676:742 For glycan labeling, a modified version of on-target derivatization with phenylhydrazine was applied. 25763881 7 48 theme potato 1008:1013 arg1 species 1015:1021 potato species 1008:1021 potato species 1008:1021 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 2 49 theme Xyl 339:341 arg1 Fuc 351:353 Fuc 351:353 Fuc 351:353 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 2 49 theme Xyl 339:341 arg1 GlcNAc2 343:349 composition Man3(Xyl)GlcNAc2 322:349 composition Man3(Xyl)GlcNAc2(Fuc) 322:354 Patatin is the most abundant glycoprotein in potato and has been described to have an oligosaccharide of composition Man3(Xyl)GlcNAc2(Fuc). 25763881 7 50 theme patatins 962:969 arg1 glycosylation 938:950 the glycosylation 934:950 the glycosylation of potato patatins 934:969 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 4 51 theme ionization 625:634 arg1 spectrometry 641:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 7 52 theme specific 1051:1058 arg1 epitopes 1067:1074 specific glycan epitopes 1051:1074 specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 1051:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 1 53 theme potential 133:141 arg1 oligosaccharides 104:119 oligosaccharides 104:119 oligosaccharides 104:119 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 1 53 theme potential 133:141 arg1 source 143:148 another potential source 125:148 another potential source of immunological cross-reaction between different plant allergens 125:214 It is hypothesized that oligosaccharides are another potential source of immunological cross-reaction between different plant allergens. 25763881 4 54 theme desorption 614:623 arg1 spectrometry 641:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25763881 0 55 theme Solanum 65:71 arg1 genus 73:77 Solanum genus 65:77 Solanum genus 65:77 N-glycome profiling of patatins from different potato species of Solanum genus. 25763881 7 56 gly glycosylation 938:950 arg1 patatins 962:969 potato patatins 955:969 potato patatins 955:969 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 7 57 theme proteins 1171:1178 arg1 manufacture 1144:1154 the manufacture 1140:1154 the manufacture of therapeutic proteins 1140:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 7 58 theme species 1015:1021 arg1 choice 998:1003 correct choice 990:1003 correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins 990:1178 This knowledge about the glycosylation of potato patatins may be helpful for correct choice of potato species to decrease the presence of specific glycan epitopes causing food allergy as well as for utilization of potatoes for the manufacture of therapeutic proteins. 25763881 4 59 theme mass 636:639 arg1 spectrometry 641:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 matrix-assisted laser desorption ionization mass spectrometry 592:652 Oligosaccharides were released by enzymatic digestion with PNAGase A and analyzed primarily by matrix-assisted laser desorption ionization mass spectrometry. 25164811 8 0 theme family 1163:1168 arg1 β-glucosidases 1172:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 10 1 theme P. 1416:1417 arg1 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 1 theme P. 1416:1417 arg1 enzyme 1445:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 2 2 theme adjacent 376:383 arg1 molecules 385:393 two adjacent molecules 372:393 two adjacent molecules in dimers and short oligomers of glucose 372:434 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 0 3 theme β-glucosidase 73:85 arg1 Cel3A 88:92 Cel3A 88:92 Cel3A from Hypocrea jecorina 88:115 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 0 3 theme β-glucosidase 73:85 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization 0:27 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 0 3 theme β-glucosidase 73:85 arg1 structures 41:50 crystal structures 33:50 crystal structures 33:50 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 11 4 theme intermolecular 1537:1550 arg1 contacts 1552:1559 intermolecular contacts 1537:1559 intermolecular contacts in the structures derived from either host 1537:1602 The glycosylations are involved in intermolecular contacts in the structures derived from either host. 25164811 7 5 theme active 1049:1054 arg1 site 1056:1059 the active site 1045:1059 the active site 1045:1059 Here, we present x-ray structures of HjCel3A with and without glucose bound in the active site. 25164811 10 6 theme jecorina-produced 1307:1323 arg1 HjCel3A 1325:1331 H. jecorina-produced HjCel3A 1304:1331 H. jecorina-produced HjCel3A 1304:1331 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 12 7 theme crystal 1692:1698 arg1 forms 1700:1704 different crystal forms 1682:1704 different crystal forms for the two protein forms 1682:1730 Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms. 25164811 4 8 theme HjCel3A 601:607 arg1 characterization 581:596 Biochemical characterization 569:596 Biochemical characterization of HjCel3A 569:607 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 10 9 theme H. 1304:1305 arg1 HjCel3A 1325:1331 H. jecorina-produced HjCel3A 1304:1331 H. jecorina-produced HjCel3A 1304:1331 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 6 10 link N-linked 942:949 arg1 glycosylation 951:963 N-linked glycosylation 942:963 N-linked glycosylation 942:963 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 9 11 theme N-linked 1251:1258 arg1 glycosylations 1260:1273 N-linked glycosylations 1251:1273 N-linked glycosylations 1251:1273 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 4 12 theme -β-D-linked 697:707 arg1 disaccharides 709:721 (1,6)-β-D-linked disaccharides 692:721 (1,6)-β-D-linked disaccharides 692:721 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 12 13 theme different 1682:1690 arg1 forms 1700:1704 different crystal forms 1682:1704 different crystal forms for the two protein forms 1682:1730 Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms. 25164811 2 14 theme Hypocrea 303:310 arg1 jecorina 312:319 the mesophilic fungus Hypocrea jecorina 281:319 the mesophilic fungus Hypocrea jecorina 281:319 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 9 15 contain have 1246:1249 arg2 glycosylations 1260:1273 N-linked glycosylations 1251:1273 N-linked glycosylations 1251:1273 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 9 15 contain have 1246:1249 arg1 structures 1230:1239 HjCel3A structures 1222:1239 HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310) 1222:1298 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 4 16 link -β-D-linked 697:707 arg1 disaccharides 709:721 (1,6)-β-D-linked disaccharides 692:721 (1,6)-β-D-linked disaccharides 692:721 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 1 17 from jecorina 151:158 arg1 mixtures 128:135 Cellulase mixtures 118:135 Cellulase mixtures from Hypocrea jecorina 118:158 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 2 18 theme fungus 296:301 arg1 jecorina 312:319 the mesophilic fungus Hypocrea jecorina 281:319 the mesophilic fungus Hypocrea jecorina 281:319 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 11 19 attach derived 1579:1585 arg1 host 1599:1602 either host 1592:1602 either host 1592:1602 The glycosylations are involved in intermolecular contacts in the structures derived from either host. 25164811 11 19 attach derived 1579:1585 arg2 structures 1568:1577 the structures 1564:1577 the structures derived from either host 1564:1602 The glycosylations are involved in intermolecular contacts in the structures derived from either host. 25164811 0 20 theme Hypocrea 99:106 arg1 jecorina 108:115 Hypocrea jecorina 99:115 Hypocrea jecorina 99:115 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 8 21 theme glycoside 1143:1151 arg1 β-glucosidases 1172:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 10 22 theme single 1336:1341 arg1 present 1366:1372 present 1366:1372 present 1366:1372 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 22 theme single 1336:1341 arg1 N-acetylglucosamine 1343:1361 a single N-acetylglucosamine 1334:1361 a single N-acetylglucosamine 1334:1361 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 2 23 theme mesophilic 285:294 arg1 jecorina 312:319 the mesophilic fungus Hypocrea jecorina 281:319 the mesophilic fungus Hypocrea jecorina 281:319 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 9 24 link N-linked 1251:1258 arg1 glycosylations 1260:1273 N-linked glycosylations 1251:1273 N-linked glycosylations 1251:1273 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 2 25 theme short 409:413 arg1 oligomers 415:423 short oligomers 409:423 short oligomers of glucose 409:434 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 10 26 from present 1366:1372 arg1 HjCel3A 1325:1331 H. jecorina-produced HjCel3A 1304:1331 H. jecorina-produced HjCel3A 1304:1331 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 6 27 contain has 919:921 arg2 degree 932:937 a higher degree 923:937 a higher degree of N-linked glycosylation 923:963 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 6 27 contain has 919:921 arg1 Pp-HjCel3A 908:917 Pp-HjCel3A 908:917 Pp-HjCel3A 908:917 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 8 28 contain have 1077:1080 arg2 architecture 1097:1108 a three-domain architecture 1082:1108 a three-domain architecture 1082:1108 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 8 28 contain have 1077:1080 arg1 structures 1066:1075 The structures 1062:1075 The structures 1062:1075 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 5 29 theme Pichia 808:813 arg1 Pp-HjCel3A 825:834 Pp-HjCel3A 825:834 Pp-HjCel3A 825:834 For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). 25164811 5 29 theme Pichia 808:813 arg1 pastoris 815:822 Pichia pastoris 808:822 Pichia pastoris (Pp-HjCel3A) 808:835 For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). 25164811 0 30 theme Biochemical 0:10 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization 0:27 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 0 30 theme Biochemical 0:10 arg1 Cel3A 88:92 Cel3A 88:92 Cel3A from Hypocrea jecorina 88:115 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 4 31 theme Biochemical 569:579 arg1 characterization 581:596 Biochemical characterization 569:596 Biochemical characterization of HjCel3A 569:607 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 6 32 theme HjCel3A 871:877 arg1 same 902:905 same 902:905 same 902:905 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 6 32 theme HjCel3A 871:877 arg1 thermostabilities 850:866 the thermostabilities 846:866 the thermostabilities of HjCel3A and Pp-HjCel3A 846:892 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 9 33 theme production 1193:1202 arg1 hosts 1204:1208 Both production hosts 1188:1208 Both production hosts 1188:1208 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 2 34 from β-glucosidase 264:276 arg1 jecorina 312:319 the mesophilic fungus Hypocrea jecorina 281:319 the mesophilic fungus Hypocrea jecorina 281:319 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 0 35 theme crystal 33:39 arg1 structures 41:50 crystal structures 33:50 crystal structures 33:50 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 8 36 theme three-domain 1084:1095 arg1 architecture 1097:1108 a three-domain architecture 1082:1108 a three-domain architecture 1082:1108 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 2 37 from molecules 385:393 arg1 dimers 398:403 dimers 398:403 dimers 398:403 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 2 37 from molecules 385:393 arg1 oligomers 415:423 short oligomers 409:423 short oligomers of glucose 409:434 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 5 38 theme H. 782:783 arg1 HjCel3A 795:801 HjCel3A 795:801 HjCel3A 795:801 For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). 25164811 5 38 theme H. 782:783 arg1 jecorina 785:792 H. jecorina 782:792 H. jecorina (HjCel3A) 782:802 For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). 25164811 11 39 gly glycosylations 1506:1519 arg1 contacts 1552:1559 intermolecular contacts 1537:1559 intermolecular contacts in the structures derived from either host 1537:1602 The glycosylations are involved in intermolecular contacts in the structures derived from either host. 25164811 7 40 theme HjCel3A 1003:1009 arg1 structures 989:998 x-ray structures 983:998 x-ray structures of HjCel3A with and without glucose bound in the active site 983:1059 Here, we present x-ray structures of HjCel3A with and without glucose bound in the active site. 25164811 4 41 theme 1,6 693:695 arg1 disaccharides 709:721 (1,6)-β-D-linked disaccharides 692:721 (1,6)-β-D-linked disaccharides 692:721 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 1 42 theme cellulose 206:214 arg1 saccharification 186:201 the saccharification 182:201 the saccharification of cellulose in biotechnical applications 182:243 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 5 43 theme crystallization 728:742 arg1 studies 744:750 crystallization studies 728:750 crystallization studies 728:750 For crystallization studies, HjCel3A was produced in both H. jecorina (HjCel3A) and Pichia pastoris (Pp-HjCel3A). 25164811 3 44 theme cellulase 502:510 arg1 mixtures 512:519 H. jecorina cellulase mixtures 490:519 H. jecorina cellulase mixtures 490:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 7 45 theme x-ray 983:987 arg1 structures 989:998 x-ray structures 983:998 x-ray structures of HjCel3A with and without glucose bound in the active site 983:1059 Here, we present x-ray structures of HjCel3A with and without glucose bound in the active site. 25164811 3 46 theme enhanced 460:467 arg1 levels 469:474 enhanced levels 460:474 enhanced levels of HjCel3A in H. jecorina cellulase mixtures 460:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 11 47 from contacts 1552:1559 arg1 structures 1568:1577 the structures 1564:1577 the structures derived from either host 1564:1602 The glycosylations are involved in intermolecular contacts in the structures derived from either host. 25164811 2 48 theme glucose 428:434 arg1 dimers 398:403 dimers 398:403 dimers 398:403 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 2 48 theme glucose 428:434 arg1 oligomers 415:423 short oligomers 409:423 short oligomers of glucose 409:434 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 8 49 theme other 1137:1141 arg1 β-glucosidases 1172:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 10 50 from HjCel3A 1325:1331 arg1 present 1366:1372 present 1366:1372 present 1366:1372 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 50 from HjCel3A 1325:1331 arg1 N-acetylglucosamine 1343:1361 a single N-acetylglucosamine 1334:1361 a single N-acetylglucosamine 1334:1361 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 6 51 theme higher 925:930 arg1 degree 932:937 a higher degree 923:937 a higher degree of N-linked glycosylation 923:963 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 12 52 theme protein 1718:1724 arg1 forms 1726:1730 the two protein forms 1710:1730 the two protein forms 1710:1730 Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms. 25164811 1 53 theme biotechnical 219:230 arg1 applications 232:243 biotechnical applications 219:243 biotechnical applications 219:243 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 0 54 theme fungal 57:62 arg1 β-glucosidase 73:85 a fungal family 3 β-glucosidase 55:85 a fungal family 3 β-glucosidase 55:85 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 6 55 theme Pp-HjCel3A 883:892 arg1 same 902:905 same 902:905 same 902:905 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 6 55 theme Pp-HjCel3A 883:892 arg1 thermostabilities 850:866 the thermostabilities 846:866 the thermostabilities of HjCel3A and Pp-HjCel3A 846:892 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 10 56 theme glycan 1457:1462 arg1 chains 1464:1469 the glycan chains 1453:1469 the glycan chains 1453:1469 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 3 57 from levels 469:474 arg1 mixtures 512:519 H. jecorina cellulase mixtures 490:519 H. jecorina cellulase mixtures 490:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 3 58 theme jecorina 493:500 arg1 mixtures 512:519 H. jecorina cellulase mixtures 490:519 H. jecorina cellulase mixtures 490:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 8 59 theme hydrolase 1153:1161 arg1 β-glucosidases 1172:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 other glycoside hydrolase family 3 β-glucosidases 1137:1185 The structures have a three-domain architecture as observed previously for other glycoside hydrolase family 3 β-glucosidases. 25164811 10 60 located present 1366:1372 arg2 N-acetylglucosamine 1343:1361 a single N-acetylglucosamine 1334:1361 a single N-acetylglucosamine 1334:1361 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 60 located present 1366:1372 arg2 present 1366:1372 present 1366:1372 present 1366:1372 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 60 located present 1366:1372 arg1 HjCel3A 1325:1331 H. jecorina-produced HjCel3A 1304:1331 H. jecorina-produced HjCel3A 1304:1331 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 60 located present 1366:1372 arg1 sites 1382:1386 both sites 1377:1386 both sites 1377:1386 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 61 theme HjCel3A 1437:1443 arg1 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 61 theme HjCel3A 1437:1443 arg1 enzyme 1445:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 0 62 from jecorina 108:115 arg1 characterization 12:27 Biochemical characterization 0:27 Biochemical characterization 0:27 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 0 62 from jecorina 108:115 arg1 Cel3A 88:92 Cel3A 88:92 Cel3A from Hypocrea jecorina 88:115 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 1 63 theme Cellulase 118:126 arg1 mixtures 128:135 Cellulase mixtures 118:135 Cellulase mixtures from Hypocrea jecorina 118:158 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 12 64 theme different 1616:1624 arg1 sizes 1626:1630 the different sizes 1612:1630 the different sizes of the glycosylations 1612:1652 Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms. 25164811 1 65 used used 173:176 arg2 mixtures 128:135 Cellulase mixtures 118:135 Cellulase mixtures from Hypocrea jecorina 118:158 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 10 66 theme pastoris-produced 1419:1435 arg1 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 Pp-HjCel3A 1400:1409 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 66 theme pastoris-produced 1419:1435 arg1 enzyme 1445:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 the P. pastoris-produced HjCel3A enzyme 1412:1450 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 4 67 dep hydrolyzes 643:652 arg1 1,4 655:657 1,4 655:657 1,4 655:657 Biochemical characterization of HjCel3A shows that the enzyme efficiently hydrolyzes (1,4)- as well as (1,2)-, (1,3)-, and (1,6)-β-D-linked disaccharides. 25164811 3 68 theme cellulose 547:555 arg1 conversion 533:542 the conversion 529:542 the conversion of cellulose to glucose 529:566 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 12 69 theme glycosylations 1639:1652 arg1 sizes 1626:1630 the different sizes 1612:1630 the different sizes of the glycosylations 1612:1652 Due to the different sizes of the glycosylations, the interactions result in different crystal forms for the two protein forms. 25164811 3 70 theme H. 490:491 arg1 mixtures 512:519 H. jecorina cellulase mixtures 490:519 H. jecorina cellulase mixtures 490:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 3 71 theme HjCel3A 479:485 arg1 levels 469:474 enhanced levels 460:474 enhanced levels of HjCel3A in H. jecorina cellulase mixtures 460:519 It has been shown that enhanced levels of HjCel3A in H. jecorina cellulase mixtures benefit the conversion of cellulose to glucose. 25164811 6 72 theme glycosylation 951:963 arg1 degree 932:937 a higher degree 923:937 a higher degree of N-linked glycosylation 923:963 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 10 73 from sites 1382:1386 arg1 present 1366:1372 present 1366:1372 present 1366:1372 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 73 from sites 1382:1386 arg1 N-acetylglucosamine 1343:1361 a single N-acetylglucosamine 1334:1361 a single N-acetylglucosamine 1334:1361 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 1 74 from saccharification 186:201 arg1 applications 232:243 biotechnical applications 219:243 biotechnical applications 219:243 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 1 75 theme Hypocrea 142:149 arg1 jecorina 151:158 Hypocrea jecorina 142:158 Hypocrea jecorina 142:158 Cellulase mixtures from Hypocrea jecorina are commonly used for the saccharification of cellulose in biotechnical applications. 25164811 6 76 theme N-linked 942:949 arg1 glycosylation 951:963 N-linked glycosylation 942:963 N-linked glycosylation 942:963 Whereas the thermostabilities of HjCel3A and Pp-HjCel3A are the same, Pp-HjCel3A has a higher degree of N-linked glycosylation. 25164811 0 77 theme family 64:69 arg1 β-glucosidase 73:85 a fungal family 3 β-glucosidase 55:85 a fungal family 3 β-glucosidase 55:85 Biochemical characterization and crystal structures of a fungal family 3 β-glucosidase, Cel3A from Hypocrea jecorina. 25164811 9 78 theme HjCel3A 1222:1228 arg1 structures 1230:1239 HjCel3A structures 1222:1239 HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310) 1222:1298 Both production hosts resulted in HjCel3A structures that have N-linked glycosylations at Asn(208) and Asn(310). 25164811 2 79 theme abundant 255:262 arg1 β-glucosidase 264:276 The most abundant β-glucosidase 246:276 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina 246:319 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 2 79 theme abundant 255:262 arg1 HjCel3A 324:330 HjCel3A 324:330 HjCel3A 324:330 The most abundant β-glucosidase in the mesophilic fungus Hypocrea jecorina is HjCel3A, which hydrolyzes the β-linkage between two adjacent molecules in dimers and short oligomers of glucose. 25164811 10 80 attach present 1366:1372 arg2 N-acetylglucosamine 1343:1361 a single N-acetylglucosamine 1334:1361 a single N-acetylglucosamine 1334:1361 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 80 attach present 1366:1372 arg2 present 1366:1372 present 1366:1372 present 1366:1372 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 80 attach present 1366:1372 arg1 HjCel3A 1325:1331 H. jecorina-produced HjCel3A 1304:1331 H. jecorina-produced HjCel3A 1304:1331 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 25164811 10 80 attach present 1366:1372 arg1 sites 1382:1386 both sites 1377:1386 both sites 1377:1386 In H. jecorina-produced HjCel3A, a single N-acetylglucosamine is present at both sites, whereas in Pp-HjCel3A, the P. pastoris-produced HjCel3A enzyme, the glycan chains consist of 8 or 4 saccharides. 27428197 3 0 theme community-based 695:709 arg1 Chinese 715:721 701 community-based Han Chinese 691:721 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 691:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 7 1 theme further 1560:1566 arg1 study 1568:1572 further study 1560:1572 further study 1560:1572 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 4 2 theme variance 1006:1013 arg1 variance 1006:1013 the variance 1002:1013 the variance in chronological age in this population 1002:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 2 theme variance 1006:1013 arg1 % 997:997 23.3% to 45.4% 984:997 23.3% to 45.4% of the variance in chronological age in this population 984:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 5 3 theme features 1105:1112 arg1 combinations 1082:1093 these combinations 1076:1093 these combinations of glycan features 1076:1112 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 1 4 theme immunoglobulin 256:269 arg1 G 271:271 immunoglobulin G 256:271 immunoglobulin G (IgG) molecules 256:287 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 4 theme immunoglobulin 256:269 arg1 IgG 274:276 IgG 274:276 IgG 274:276 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 0 5 theme ages 79:82 arg1 biomarker 37:45 potential biomarker 27:45 potential biomarker of chronological and biological ages 27:82 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 1 6 theme inflammatory 327:338 arg1 cascade 340:346 the inflammatory cascade 323:346 the inflammatory cascade 323:346 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 7 from steps 314:318 arg1 essential 296:304 essential 296:304 essential 296:304 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 8 theme important 144:152 arg1 process 181:187 an important post-translation modifying process 141:187 an important post-translation modifying process 141:187 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 8 theme important 144:152 arg1 function 244:251 function 244:251 function 244:251 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 8 theme important 144:152 arg1 structure 230:238 structure 230:238 structure 230:238 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 4 9 theme specific 925:932 arg1 combinations 934:945 specific combinations 925:945 specific combinations of these glycan features 925:970 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 1 10 theme G 271:271 arg1 molecules 279:287 immunoglobulin G (IgG) molecules 256:287 immunoglobulin G (IgG) molecules 256:287 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 0 11 theme community-based 87:101 arg1 study 103:107 A community-based study 85:107 A community-based study in a Han Chinese population 85:135 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 1 12 theme cascade 340:346 arg1 steps 314:318 many steps 309:318 many steps of the inflammatory cascade 309:346 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 13 from essential 296:304 arg1 steps 314:318 many steps 309:318 many steps of the inflammatory cascade 309:346 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 5 14 theme more 1122:1125 arg1 information 1138:1148 more predictive information 1122:1148 more predictive information than other single markers of biological age such as telomere length 1122:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 7 15 theme IgG 1421:1423 arg1 glycosylation 1425:1437 IgG glycosylation 1421:1437 IgG glycosylation 1421:1437 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 3 16 from glycosylation 623:635 arg1 patterns 596:603 various patterns 588:603 various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 588:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 3 17 from patterns 596:603 arg1 glycosylation 623:635 IgG glycosylation 619:635 IgG glycosylation 619:635 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 4 18 from % 997:997 arg1 age 1032:1034 chronological age 1018:1034 chronological age in this population 1018:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 5 19 theme age 1190:1192 arg1 length 1211:1216 telomere length 1202:1216 telomere length 1202:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 5 19 theme age 1190:1192 arg1 markers 1168:1174 other single markers 1155:1174 other single markers of biological age such as telomere length 1155:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 6 20 theme biological 1330:1339 arg1 age 1341:1343 biological age 1330:1343 biological age 1330:1343 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 2 21 dep demonstrated 362:373 arg1 whereas 523:529 whereas 523:529 whereas 523:529 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 4 22 theme % 988:988 arg1 variance 1006:1013 the variance 1002:1013 the variance in chronological age in this population 1002:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 22 theme % 988:988 arg1 % 997:997 23.3% to 45.4% 984:997 23.3% to 45.4% of the variance in chronological age in this population 984:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 1 23 theme post-translation 154:169 arg1 process 181:187 an important post-translation modifying process 141:187 an important post-translation modifying process 141:187 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 23 theme post-translation 154:169 arg1 function 244:251 function 244:251 function 244:251 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 23 theme post-translation 154:169 arg1 structure 230:238 structure 230:238 structure 230:238 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 7 24 theme biological 1488:1497 arg1 ages 1499:1502 both chronological and biological ages 1465:1502 both chronological and biological ages 1465:1502 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 3 25 theme Han 711:713 arg1 Chinese 715:721 701 community-based Han Chinese 691:721 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 691:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 1 26 gly glycosylation 190:202 arg1 molecules 279:287 immunoglobulin G (IgG) molecules 256:287 immunoglobulin G (IgG) molecules 256:287 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 3 27 from changes 608:614 arg1 glycosylation 623:635 IgG glycosylation 619:635 IgG glycosylation 619:635 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 1 28 theme modifying 171:179 arg1 process 181:187 an important post-translation modifying process 141:187 an important post-translation modifying process 141:187 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 28 theme modifying 171:179 arg1 function 244:251 function 244:251 function 244:251 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 28 theme modifying 171:179 arg1 structure 230:238 structure 230:238 structure 230:238 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 0 29 theme IgG 10:12 arg1 N-glycans 14:22 IgG N-glycans 10:22 IgG N-glycans 10:22 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 2 30 theme European 501:508 arg1 populations 510:520 several European populations 493:520 several European populations 493:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 4 31 dep change 892:897 arg1 BG1 834:836 BG1 834:836 BG1 834:836 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 A2G1 802:805 A2G1 802:805 A2G1 802:805 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 FA2[3 839:843 FA2[3 839:843 FA2[3 839:843 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 FA2[6 808:812 FA2[6 808:812 FA2[6 808:812 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 G1 824:825 G1 824:825 G1 824:825 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 glycans 777:783 Eleven IgG glycans 766:783 Eleven IgG glycans 766:783 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 FA2[6 828:832 FA2[6 828:832 FA2[6 828:832 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 BG1 845:847 BG1 845:847 BG1 845:847 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 G1 814:815 G1 814:815 G1 814:815 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 31 dep change 892:897 arg1 FA2[3 818:822 FA2[3 818:822 FA2[3 818:822 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 5 32 theme telomere 1202:1209 arg1 length 1211:1216 telomere length 1202:1216 telomere length 1202:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 6 33 theme aspartate 1287:1295 arg1 aminotransferase 1297:1312 aspartate aminotransferase 1287:1312 aspartate aminotransferase 1287:1312 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 5 34 theme single 1161:1166 arg1 length 1211:1216 telomere length 1202:1216 telomere length 1202:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 5 34 theme single 1161:1166 arg1 markers 1168:1174 other single markers 1155:1174 other single markers of biological age such as telomere length 1155:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 6 35 theme plasma 1268:1273 arg1 glucose 1275:1281 fasting plasma glucose 1260:1281 fasting plasma glucose 1260:1281 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 1 36 theme molecules 279:287 arg1 function 244:251 function 244:251 function 244:251 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 36 theme molecules 279:287 arg1 process 181:187 an important post-translation modifying process 141:187 an important post-translation modifying process 141:187 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 1 36 theme molecules 279:287 arg1 structure 230:238 structure 230:238 structure 230:238 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 0 37 theme Han 114:116 arg1 population 126:135 a Han Chinese population 112:135 a Han Chinese population 112:135 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 4 38 theme features 963:970 arg1 combinations 934:945 specific combinations 925:945 specific combinations of these glycan features 925:970 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 2 39 theme IgG 424:426 arg1 component 433:441 a component 431:441 a component of predictive biomarkers for chronological age in several European populations 431:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 2 39 theme IgG 424:426 arg1 features 412:419 glycosylation features 398:419 glycosylation features of IgG 398:426 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 6 40 theme fasting 1260:1266 arg1 glucose 1275:1281 fasting plasma glucose 1260:1281 fasting plasma glucose 1260:1281 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 6 41 theme combined 1378:1385 arg1 features 1394:1401 the combined glycan features 1374:1401 the combined glycan features 1374:1401 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 2 42 theme chronological 472:484 arg1 age 486:488 chronological age 472:488 chronological age in several European populations 472:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 7 43 theme possible 1518:1525 arg1 role 1527:1530 its possible role 1514:1530 its possible role in the aging process 1514:1551 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 2 44 from age 486:488 arg1 populations 510:520 several European populations 493:520 several European populations 493:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 7 45 from role 1527:1530 arg1 process 1545:1551 the aging process 1535:1551 the aging process 1535:1551 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 5 46 theme biological 1179:1188 arg1 age 1190:1192 biological age 1179:1192 biological age 1179:1192 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 0 47 theme potential 27:35 arg1 biomarker 37:45 potential biomarker 27:45 potential biomarker of chronological and biological ages 27:82 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 6 48 theme clinical 1236:1243 arg1 aminotransferase 1297:1312 aspartate aminotransferase 1287:1312 aspartate aminotransferase 1287:1312 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 6 48 theme clinical 1236:1243 arg1 glucose 1275:1281 fasting plasma glucose 1260:1281 fasting plasma glucose 1260:1281 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 6 48 theme clinical 1236:1243 arg1 traits 1245:1250 the clinical traits 1232:1250 the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age 1232:1343 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 0 49 theme Chinese 118:124 arg1 population 126:135 a Han Chinese population 112:135 a Han Chinese population 112:135 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 1 50 dep structure 230:238 arg1 the 226:228 the 226:228 the 226:228 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 3 51 theme various 588:594 arg1 patterns 596:603 various patterns 588:603 various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 588:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 2 52 theme glycosylation 398:410 arg1 component 433:441 a component 431:441 a component of predictive biomarkers for chronological age in several European populations 431:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 2 52 theme glycosylation 398:410 arg1 features 412:419 glycosylation features 398:419 glycosylation features of IgG 398:426 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 0 53 theme chronological 50:62 arg1 ages 79:82 chronological and biological ages 50:82 chronological and biological ages 50:82 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 3 54 theme old 760:762 arg1 years 754:758 23-68 years 748:758 23-68 years old 748:762 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 2 55 theme several 493:499 arg1 populations 510:520 several European populations 493:520 several European populations 493:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 4 56 from age 1032:1034 arg1 variance 1006:1013 the variance 1002:1013 the variance in chronological age in this population 1002:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 56 from age 1032:1034 arg1 population 1044:1053 this population 1039:1053 this population 1039:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 56 from age 1032:1034 arg1 % 997:997 23.3% to 45.4% 984:997 23.3% to 45.4% of the variance in chronological age in this population 984:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 57 theme chronological 1018:1030 arg1 age 1032:1034 chronological age 1018:1034 chronological age in this population 1018:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 2 58 theme predictive 446:455 arg1 biomarkers 457:466 predictive biomarkers 446:466 predictive biomarkers for chronological age in several European populations 446:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 1 59 theme many 309:312 arg1 steps 314:318 many steps 309:318 many steps of the inflammatory cascade 309:346 As an important post-translation modifying process, glycosylation significantly affects the structure and function of immunoglobulin G (IgG) molecules and is essential in many steps of the inflammatory cascade. 27428197 3 60 theme IgG 670:672 arg1 glycosylation 674:686 IgG glycosylation 670:686 IgG glycosylation 670:686 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 3 61 theme IgG 619:621 arg1 glycosylation 623:635 IgG glycosylation 619:635 IgG glycosylation 619:635 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 5 62 theme predictive 1127:1136 arg1 information 1138:1148 more predictive information 1122:1148 more predictive information than other single markers of biological age such as telomere length 1122:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 3 63 gly glycosylation 674:686 arg1 Chinese 715:721 701 community-based Han Chinese 691:721 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 691:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 4 64 dep A2BG2 856:860 arg1 FA2G2S1 870:876 FA2G2S1 870:876 FA2G2S1 870:876 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 64 dep A2BG2 856:860 arg1 FA2G2S2 883:889 FA2G2S2 883:889 FA2G2S2 883:889 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 64 dep A2BG2 856:860 arg1 FA2G2 863:867 FA2G2 863:867 FA2G2 863:867 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 0 65 from study 103:107 arg1 population 126:135 a Han Chinese population 112:135 a Han Chinese population 112:135 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 3 66 theme changes 608:614 arg1 patterns 596:603 various patterns 588:603 various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old) 588:763 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 7 67 theme aging 1539:1543 arg1 process 1545:1551 the aging process 1535:1551 the aging process 1535:1551 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 7 68 dep appears 1439:1445 arg1 correlate 1450:1458 correlate 1450:1458 appears to correlate with both chronological and biological ages 1439:1502 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 4 69 from variance 1006:1013 arg1 age 1032:1034 chronological age 1018:1034 chronological age in this population 1018:1053 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 4 70 dep % 997:997 arg1 to 990:991 to 990:991 to 990:991 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 6 71 theme glycan 1387:1392 arg1 features 1394:1401 the combined glycan features 1374:1401 the combined glycan features 1374:1401 In addition, the clinical traits such as fasting plasma glucose and aspartate aminotransferase associated with biological age are strongly correlated with the combined glycan features. 27428197 4 72 theme glycan 956:961 arg1 features 963:970 these glycan features 950:970 these glycan features 950:970 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 7 73 theme chronological 1470:1482 arg1 ages 1499:1502 both chronological and biological ages 1465:1502 both chronological and biological ages 1465:1502 We conclude that IgG glycosylation appears to correlate with both chronological and biological ages, and thus its possible role in the aging process merits further study. 27428197 0 74 theme biological 68:77 arg1 ages 79:82 chronological and biological ages 50:82 chronological and biological ages 50:82 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 4 75 theme IgG 773:775 arg1 glycans 777:783 Eleven IgG glycans 766:783 Eleven IgG glycans 766:783 Eleven IgG glycans, including FA2B, A2G1, FA2[6]G1, FA2[3]G1, FA2[6]BG1, FA2[3]BG1, A2G2, A2BG2, FA2G2, FA2G2S1, and FA2G2S2, change considerably with age and specific combinations of these glycan features can explain 23.3% to 45.4% of the variance in chronological age in this population. 27428197 5 76 theme other 1155:1159 arg1 length 1211:1216 telomere length 1202:1216 telomere length 1202:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 5 76 theme other 1155:1159 arg1 markers 1168:1174 other single markers 1155:1174 other single markers of biological age such as telomere length 1155:1216 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 2 77 theme biomarkers 457:466 arg1 component 433:441 a component 431:441 a component of predictive biomarkers for chronological age in several European populations 431:520 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 2 77 theme biomarkers 457:466 arg1 features 412:419 glycosylation features 398:419 glycosylation features of IgG 398:426 Studies have demonstrated the potential of using glycosylation features of IgG as a component of predictive biomarkers for chronological age in several European populations, whereas no study has been reported in Chinese. 27428197 3 78 dep males 728:732 arg1 years 754:758 23-68 years 748:758 23-68 years old 748:762 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 0 79 dep Profiling 0:8 arg1 study 103:107 A community-based study 85:107 A community-based study in a Han Chinese population 85:135 Profiling IgG N-glycans as potential biomarker of chronological and biological ages: A community-based study in a Han Chinese population. 27428197 5 80 theme glycan 1098:1103 arg1 features 1105:1112 glycan features 1098:1112 glycan features 1098:1112 This indicates that these combinations of glycan features provide more predictive information than other single markers of biological age such as telomere length. 27428197 3 81 dep Chinese 715:721 arg1 males 728:732 244 males 724:732 244 males 724:732 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 27428197 3 81 dep Chinese 715:721 arg1 females 739:745 457 females 735:745 457 females 735:745 Herein, we report various patterns of changes in IgG glycosylation associated with age by analyzing IgG glycosylation in 701 community-based Han Chinese (244 males, 457 females; 23-68 years old). 24975648 0 0 theme hydrolase 81:89 arg1 family 91:96 glycoside hydrolase family 2 71:98 glycoside hydrolase family 2 71:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 7 1 theme other 1131:1135 arg1 members 1137:1143 other members 1131:1143 other members of GH family 2 1131:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 2 theme GH 703:704 arg1 mannosidase 715:725 a GH family 2 mannosidase 701:725 a GH family 2 mannosidase 701:725 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 1 3 theme biomass 243:249 arg1 Hemicellulose 185:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 1 3 theme biomass 243:249 arg1 part 215:218 an important part 202:218 an important part of the plant cell wall biomass 202:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 0 4 theme glycoside 71:79 arg1 family 91:96 glycoside hydrolase family 2 71:98 glycoside hydrolase family 2 71:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 2 5 theme nonreducing 351:361 arg1 residues 363:370 nonreducing residues 351:370 nonreducing residues of β-d-mannose 351:385 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 5 theme nonreducing 351:361 arg1 β-d-mannose 375:385 β-d-mannose 375:385 β-d-mannose 375:385 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 7 6 theme β-mannoside 1195:1205 arg1 binding 1207:1213 β-mannoside binding 1195:1213 β-mannoside binding 1195:1213 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 7 theme thetaiotaomicron 764:779 arg1 enzyme 781:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 9 8 theme T. 1421:1422 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 7 9 theme binding 1207:1213 arg1 mechanism 1182:1190 the molecular mechanism 1168:1190 the molecular mechanism of β-mannoside binding and recognition 1168:1229 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 10 theme Bacteroides 752:762 arg1 enzyme 781:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 4 11 theme mannosidase 715:725 arg1 structure 688:696 The only published X-ray structure 663:696 The only published X-ray structure of a GH family 2 mannosidase 663:725 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 4 11 theme mannosidase 715:725 arg1 that 730:733 that 730:733 that 730:733 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 9 12 theme BankIt1712036 1500:1512 arg1 KJ624918 1527:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 8 13 theme DATABASE 1291:1298 arg1 Coordinates 1300:1310 DATABASE Coordinates 1291:1310 DATABASE Coordinates 1291:1310 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 9 14 theme harzianum 1424:1432 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 9 15 theme nucleotide 1451:1460 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 2 16 theme hemicellulose 411:423 arg1 polysaccharides 444:458 hemicellulose mannose-containing polysaccharides 411:458 hemicellulose mannose-containing polysaccharides 411:458 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 16 theme hemicellulose 411:423 arg1 galactomannans 481:494 galactomannans 481:494 galactomannans 481:494 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 16 theme hemicellulose 411:423 arg1 mannans 469:475 mannans 469:475 mannans 469:475 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 7 17 theme GH 1148:1149 arg1 family 1151:1156 GH family 2 1148:1158 GH family 2 1148:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 0 18 from function 32:39 arg1 family 91:96 glycoside hydrolase family 2 71:98 glycoside hydrolase family 2 71:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 7 19 theme structural 1102:1111 arg1 architecture 1113:1124 its structural architecture 1098:1124 its structural architecture with other members of GH family 2 1098:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 9 20 theme GeneMark.hmm 1514:1525 arg1 KJ624918 1527:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 1 21 theme UNLABELLED 174:183 arg1 Hemicellulose 185:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 1 21 theme UNLABELLED 174:183 arg1 part 215:218 an important part 202:218 an important part of the plant cell wall biomass 202:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 5 22 theme mannosidases 817:828 arg1 structures 792:801 No structures 789:801 No structures of eukaryotic mannosidases of this family 789:843 No structures of eukaryotic mannosidases of this family are currently available. 24975648 6 23 theme family 950:955 arg1 β-mannosidase 959:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 7 24 theme recognition 1219:1229 arg1 mechanism 1182:1190 the molecular mechanism 1168:1190 the molecular mechanism of β-mannoside binding and recognition 1168:1229 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 8 25 theme Data 1390:1393 arg1 Bank 1395:1398 the Protein Data Bank 1378:1398 the Protein Data Bank (4CVU and 4UOJ) 1378:1414 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 7 26 theme galactomannan-binding 1263:1283 arg1 site 1285:1288 its putative galactomannan-binding site 1250:1288 its putative galactomannan-binding site 1250:1288 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 27 theme X-ray 682:686 arg1 structure 688:696 The only published X-ray structure 663:696 The only published X-ray structure of a GH family 2 mannosidase 663:725 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 4 27 theme X-ray 682:686 arg1 that 730:733 that 730:733 that 730:733 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 7 28 theme molecular 1172:1180 arg1 mechanism 1182:1190 the molecular mechanism 1168:1190 the molecular mechanism of β-mannoside binding and recognition 1168:1229 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 7 29 from similarities 1082:1093 arg1 architecture 1113:1124 its structural architecture 1098:1124 its structural architecture with other members of GH family 2 1098:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 9 30 contain has 1471:1473 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 9 30 contain has 1471:1473 arg2 KJ624918 1527:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 8 31 theme Protein 1382:1388 arg1 Bank 1395:1398 the Protein Data Bank 1378:1398 the Protein Data Bank (4CVU and 4UOJ) 1378:1414 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 7 32 theme β-mannosidase 1058:1070 arg1 comparisons 1022:1032 Structural comparisons 1011:1032 Structural comparisons of the T. harzianum GH2 β-mannosidase 1011:1070 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 33 theme published 672:680 arg1 structure 688:696 The only published X-ray structure 663:696 The only published X-ray structure of a GH family 2 mannosidase 663:725 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 4 33 theme published 672:680 arg1 that 730:733 that 730:733 that 730:733 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 6 34 theme β-mannosidase 959:971 arg1 structure 912:920 the structure 908:920 the structure of Trichoderma harzianum GH family 2 β-mannosidase 908:971 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 0 35 theme crystal 118:124 arg1 structures 126:135 multiple crystal structures 109:135 multiple crystal structures of the Trichoderma harzianum enzyme 109:171 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 4 36 theme bacterial 742:750 arg1 enzyme 781:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 the bacterial Bacteroides thetaiotaomicron enzyme 738:786 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 9 37 theme GenBank 1475:1481 arg1 number 1493:1498 GenBank accession number 1475:1498 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 5 38 theme family 838:843 arg1 mannosidases 817:828 eukaryotic mannosidases 806:828 eukaryotic mannosidases of this family 806:843 No structures of eukaryotic mannosidases of this family are currently available. 24975648 1 39 theme cellulosic 271:280 arg1 technologies 290:301 cellulosic ethanol technologies 271:301 cellulosic ethanol technologies 271:301 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 0 40 theme multiple 109:116 arg1 structures 126:135 multiple crystal structures 109:135 multiple crystal structures of the Trichoderma harzianum enzyme 109:171 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 1 41 theme important 205:213 arg1 Hemicellulose 185:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose 174:197 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 1 41 theme important 205:213 arg1 part 215:218 an important part 202:218 an important part of the plant cell wall biomass 202:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 7 42 theme family 1151:1156 arg1 members 1137:1143 other members 1131:1143 other members of GH family 2 1131:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 1 43 theme ethanol 282:288 arg1 technologies 290:301 cellulosic ethanol technologies 271:301 cellulosic ethanol technologies 271:301 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 7 44 theme harzianum 1044:1052 arg1 β-mannosidase 1058:1070 the T. harzianum GH2 β-mannosidase 1037:1070 the T. harzianum GH2 β-mannosidase 1037:1070 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 8 45 theme factor 1335:1340 arg1 amplitudes 1342:1351 observed structure factor amplitudes 1316:1351 observed structure factor amplitudes 1316:1351 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 8 46 theme observed 1316:1323 arg1 amplitudes 1342:1351 observed structure factor amplitudes 1316:1351 observed structure factor amplitudes 1316:1351 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 9 47 theme accession 1483:1491 arg1 number 1493:1498 GenBank accession number 1475:1498 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 9 48 theme β-mannosidase 1434:1446 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 6 49 theme GH 947:948 arg1 β-mannosidase 959:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 7 50 theme GH2 1054:1056 arg1 β-mannosidase 1058:1070 the T. harzianum GH2 β-mannosidase 1037:1070 the T. harzianum GH2 β-mannosidase 1037:1070 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 2 51 theme mannose-containing 425:442 arg1 polysaccharides 444:458 hemicellulose mannose-containing polysaccharides 411:458 hemicellulose mannose-containing polysaccharides 411:458 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 51 theme mannose-containing 425:442 arg1 galactomannans 481:494 galactomannans 481:494 galactomannans 481:494 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 51 theme mannose-containing 425:442 arg1 mannans 469:475 mannans 469:475 mannans 469:475 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 0 52 theme fungal 44:49 arg1 β-mannosidases 51:64 fungal β-mannosidases 44:64 fungal β-mannosidases from glycoside hydrolase family 2 44:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 6 53 theme harzianum 937:945 arg1 β-mannosidase 959:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 0 54 theme Trichoderma 144:154 arg1 enzyme 166:171 the Trichoderma harzianum enzyme 140:171 the Trichoderma harzianum enzyme 140:171 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 7 55 theme Structural 1011:1020 arg1 comparisons 1022:1032 Structural comparisons 1011:1032 Structural comparisons of the T. harzianum GH2 β-mannosidase 1011:1070 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 3 56 theme hydrolase 546:554 arg1 families 561:568 glycoside hydrolase (GH) families 1, 2, and 5 536:580 glycoside hydrolase (GH) families 1, 2, and 5 536:580 β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. 24975648 5 57 theme eukaryotic 806:815 arg1 mannosidases 817:828 eukaryotic mannosidases 806:828 eukaryotic mannosidases of this family 806:843 No structures of eukaryotic mannosidases of this family are currently available. 24975648 6 58 theme Trichoderma 925:935 arg1 β-mannosidase 959:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 Trichoderma harzianum GH family 2 β-mannosidase 925:971 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 7 59 theme T. 1041:1042 arg1 β-mannosidase 1058:1070 the T. harzianum GH2 β-mannosidase 1037:1070 the T. harzianum GH2 β-mannosidase 1037:1070 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 4 60 theme family 706:711 arg1 mannosidase 715:725 a GH family 2 mannosidase 701:725 a GH family 2 mannosidase 701:725 The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. 24975648 0 61 theme enzyme 166:171 arg1 structures 126:135 multiple crystal structures 109:135 multiple crystal structures of the Trichoderma harzianum enzyme 109:171 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 0 62 dep structure 18:26 arg1 the 14:16 the 14:16 the 14:16 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 2 63 theme capable 331:337 arg1 enzymes 323:329 enzymes 323:329 enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans 323:494 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 63 theme capable 331:337 arg1 β-Mannosidases 304:317 β-Mannosidases 304:317 β-Mannosidases 304:317 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 9 64 theme 2A 1448:1449 arg1 sequence 1462:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence 1417:1469 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 0 65 theme harzianum 156:164 arg1 enzyme 166:171 the Trichoderma harzianum enzyme 140:171 the Trichoderma harzianum enzyme 140:171 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 7 66 with architecture 1113:1124 arg1 members 1137:1143 other members 1131:1143 other members of GH family 2 1131:1158 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 3 67 theme glycoside 536:544 arg1 hydrolase 546:554 glycoside hydrolase 536:554 glycoside hydrolase (GH) families 1, 2, and 5 536:580 β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. 24975648 3 67 theme glycoside 536:544 arg1 GH 557:558 GH 557:558 GH 557:558 β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. 24975648 1 68 theme plant 227:231 arg1 biomass 243:249 the plant cell wall biomass 223:249 the plant cell wall biomass 223:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 9 69 theme number 1493:1498 arg1 KJ624918 1527:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 GenBank accession number BankIt1712036 GeneMark.hmm KJ624918 1475:1534 The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. 24975648 0 70 from family 91:96 arg1 function 32:39 function 32:39 function 32:39 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 0 70 from family 91:96 arg1 β-mannosidases 51:64 fungal β-mannosidases 44:64 fungal β-mannosidases from glycoside hydrolase family 2 44:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 0 70 from family 91:96 arg1 structure 18:26 structure 18:26 structure 18:26 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 3 71 theme enzymes 609:615 arg1 handful 594:600 only a handful 587:600 only a handful of the enzymes 587:615 β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. 24975648 2 72 theme β-d-mannose 375:385 arg1 residues 363:370 nonreducing residues 351:370 nonreducing residues of β-d-mannose 351:385 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 2 72 theme β-d-mannose 375:385 arg1 β-d-mannose 375:385 β-d-mannose 375:385 β-d-mannose 375:385 β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. 24975648 8 73 dep Bank 1395:1398 arg1 4UOJ 1410:1413 4UOJ 1410:1413 4UOJ 1410:1413 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 8 73 dep Bank 1395:1398 arg1 4CVU 1401:1404 4CVU 1401:1404 4CVU 1401:1404 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 1 74 theme cell 233:236 arg1 biomass 243:249 the plant cell wall biomass 223:249 the plant cell wall biomass 223:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 24975648 0 75 theme β-mannosidases 51:64 arg1 function 32:39 function 32:39 function 32:39 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 0 75 theme β-mannosidases 51:64 arg1 structure 18:26 structure 18:26 structure 18:26 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 0 76 from structure 18:26 arg1 family 91:96 glycoside hydrolase family 2 71:98 glycoside hydrolase family 2 71:98 Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme. 24975648 6 77 theme 1.9-Å 993:997 arg1 resolution 999:1008 1.9-Å resolution 993:1008 1.9-Å resolution 993:1008 To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. 24975648 7 78 theme putative 1254:1261 arg1 site 1285:1288 its putative galactomannan-binding site 1250:1288 its putative galactomannan-binding site 1250:1288 Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. 24975648 8 79 theme structure 1325:1333 arg1 amplitudes 1342:1351 observed structure factor amplitudes 1316:1351 observed structure factor amplitudes 1316:1351 DATABASE Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). 24975648 1 80 theme wall 238:241 arg1 biomass 243:249 the plant cell wall biomass 223:249 the plant cell wall biomass 223:249 UNLABELLED Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. 28543513 9 0 theme retention 1458:1466 arg1 times 1468:1472 retention times 1458:1472 retention times 1458:1472 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 0 1 theme LC-MS/MS 0:7 arg1 profiling 18:26 LC-MS/MS isomeric profiling 0:26 LC-MS/MS isomeric profiling of permethylated N-glycans 0:53 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 9 2 theme mass 1485:1488 arg1 spectra 1490:1496 tandem mass spectra 1478:1496 tandem mass spectra 1478:1496 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 6 3 theme C18-LC-MS/MS 973:984 arg1 analysis 986:993 C18-LC-MS/MS analysis 973:993 C18-LC-MS/MS analysis 973:993 In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. 28543513 2 4 from structures 355:364 arg1 blood 381:385 blood 381:385 blood 381:385 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 4 from structures 355:364 arg1 serum 390:394 serum 390:394 serum 390:394 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 11 5 theme glycan 1837:1842 arg1 changes 1844:1850 disease prompted aberrant glycan changes 1811:1850 disease prompted aberrant glycan changes 1811:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 3 6 theme glycosylation 548:560 arg1 patterns 562:569 the glycosylation patterns 544:569 the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 544:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 5 7 theme glycan 865:870 arg1 forms 872:876 non-isomeric and isomeric permethylated glycan forms 825:876 non-isomeric and isomeric permethylated glycan forms 825:876 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 3 8 gly glycoprotein 586:597 arg1 glycoprotein 586:597 haptoglobin glycoprotein 574:597 haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 574:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 0 9 theme patients 134:141 arg1 haptoglobin 74:84 serum haptoglobin 68:84 serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients 68:141 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 1 10 theme biomarkers 225:234 arg1 identification 198:211 the identification 194:211 the identification of reliable biomarkers 194:234 Early stage detection and cancer treatment demand the identification of reliable biomarkers. 28543513 3 11 theme glycoprotein 586:597 arg1 patterns 562:569 the glycosylation patterns 544:569 the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 544:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 3 12 gly glycosylation 548:560 arg1 glycoprotein 586:597 haptoglobin glycoprotein 574:597 haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 574:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 8 13 theme early 1412:1416 arg1 HCC 1424:1426 early stage HCC 1412:1426 early stage HCC 1412:1426 The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. 28543513 11 14 theme separation 1787:1796 arg1 importance 1764:1773 the potential importance 1750:1773 the potential importance of isomeric separation for defining disease prompted aberrant glycan changes 1750:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 9 15 theme isomeric 1538:1545 arg1 structures 1554:1563 the specific isomeric glycan structures 1525:1563 the specific isomeric glycan structures 1525:1563 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 4 16 theme haptoglobin 691:701 arg1 samples 703:709 The haptoglobin samples 687:709 The haptoglobin samples 687:709 The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. 28543513 0 17 theme serum 68:72 arg1 haptoglobin 74:84 serum haptoglobin 68:84 serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients 68:141 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 5 18 theme graphitic 912:920 arg1 columns 935:941 porous graphitic carbon (PGC) columns 905:941 porous graphitic carbon (PGC) columns 905:941 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 2 19 from variation 300:308 arg1 blood 381:385 blood 381:385 blood 381:385 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 19 from variation 300:308 arg1 serum 390:394 serum 390:394 serum 390:394 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 11 20 theme prompted 1819:1826 arg1 changes 1844:1850 disease prompted aberrant glycan changes 1811:1850 disease prompted aberrant glycan changes 1811:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 6 21 theme glycan 999:1004 arg1 structures 1006:1015 25 glycan structures 996:1015 25 glycan structures 996:1015 In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. 28543513 0 22 theme carcinoma 104:112 arg1 haptoglobin 74:84 serum haptoglobin 68:84 serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients 68:141 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 10 23 gly fucosylated 1650:1660 arg1 significant 1618:1628 significant 1618:1628 significant 1618:1628 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 10 23 gly fucosylated 1650:1660 arg1 isomers 1584:1590 the glycan isomers 1573:1590 the glycan isomers 1573:1590 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 10 23 gly fucosylated 1650:1660 arg1 All 1566:1568 All 1566:1568 All 1566:1568 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 8 24 theme glycan 1374:1379 arg1 patterns 1389:1396 The glycan isoform patterns 1370:1396 The glycan isoform patterns 1370:1396 The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. 28543513 10 25 theme sialic 1690:1695 arg1 moieties 1702:1709 α-2,6 linked sialic acid moieties 1677:1709 α-2,6 linked sialic acid moieties 1677:1709 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 4 26 theme antibody-immobilized 746:765 arg1 column 767:772 an antibody-immobilized column 743:772 an antibody-immobilized column prior to the release of N-glycans 743:806 The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. 28543513 10 27 theme α-2,6 1677:1681 arg1 moieties 1702:1709 α-2,6 linked sialic acid moieties 1677:1709 α-2,6 linked sialic acid moieties 1677:1709 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 5 28 theme non-isomeric 825:836 arg1 forms 872:876 non-isomeric and isomeric permethylated glycan forms 825:876 non-isomeric and isomeric permethylated glycan forms 825:876 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 11 29 theme study 1731:1735 arg1 result 1716:1721 The result 1712:1721 The result of this study 1712:1735 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 4 30 theme prior 774:778 arg1 column 767:772 an antibody-immobilized column 743:772 an antibody-immobilized column prior to the release of N-glycans 743:806 The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. 28543513 5 31 theme isomeric 842:849 arg1 forms 872:876 non-isomeric and isomeric permethylated glycan forms 825:876 non-isomeric and isomeric permethylated glycan forms 825:876 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 12 32 theme stage 1923:1927 arg1 HCC 1929:1931 early stage HCC 1917:1931 early stage HCC 1917:1931 The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis. 28543513 5 33 theme porous 905:910 arg1 columns 935:941 porous graphitic carbon (PGC) columns 905:941 porous graphitic carbon (PGC) columns 905:941 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 7 34 theme composition 1304:1314 arg1 distributions 1267:1279 isomeric distributions 1258:1279 isomeric distributions of a particular glycan composition 1258:1314 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 0 35 theme permethylated 31:43 arg1 N-glycans 45:53 permethylated N-glycans 31:53 permethylated N-glycans 31:53 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 2 36 theme several 451:457 arg1 cancers 459:465 several cancers 451:465 several cancers 451:465 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 3 37 theme hepatocellular 646:659 arg1 carcinoma 661:669 hepatocellular carcinoma 646:669 hepatocellular carcinoma (HCC) patients 646:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 3 37 theme hepatocellular 646:659 arg1 HCC 672:674 HCC 672:674 HCC 672:674 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 8 38 theme cirrhotic 1433:1441 arg1 patients 1443:1450 cirrhotic patients 1433:1450 cirrhotic patients 1433:1450 The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. 28543513 12 39 theme isoforms 1882:1889 arg1 levels 1857:1862 The levels 1853:1862 The levels of several glycan isoforms 1853:1889 The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis. 28543513 7 40 from different 1321:1329 arg1 abundances 1334:1343 abundances 1334:1343 abundances between the two cohorts 1334:1367 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 7 41 theme glycan 1297:1302 arg1 composition 1304:1314 a particular glycan composition 1284:1314 a particular glycan composition 1284:1314 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 7 42 from abundances 1334:1343 arg1 different 1321:1329 different 1321:1329 different 1321:1329 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 1 43 theme stage 150:154 arg1 detection 156:164 Early stage detection 144:164 Early stage detection 144:164 Early stage detection and cancer treatment demand the identification of reliable biomarkers. 28543513 3 44 theme LC-MS/MS-based 496:509 arg1 analysis 511:518 an LC-MS/MS-based analysis 493:518 an LC-MS/MS-based analysis 493:518 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 2 45 theme glycan 348:353 arg1 structures 355:364 the glycan structures 344:364 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 10 46 link linked 1683:1688 arg1 moieties 1702:1709 α-2,6 linked sialic acid moieties 1677:1709 α-2,6 linked sialic acid moieties 1677:1709 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 1 47 theme cancer 170:175 arg1 treatment 177:185 cancer treatment 170:185 cancer treatment 170:185 Early stage detection and cancer treatment demand the identification of reliable biomarkers. 28543513 10 48 dep either 1636:1641 arg1 fucosylated 1650:1660 fucosylated 1650:1660 fucosylated 1650:1660 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 2 49 theme glycosylation 313:325 arg1 level 327:331 glycosylation level 313:331 glycosylation level 313:331 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 9 50 theme tandem 1478:1483 arg1 spectra 1490:1496 tandem mass spectra 1478:1496 tandem mass spectra 1478:1496 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 6 51 theme analysis 986:993 arg1 case 965:968 the case 961:968 the case of C18-LC-MS/MS analysis 961:993 In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. 28543513 0 52 theme cirrhotic 124:132 arg1 patients 134:141 cirrhotic patients 124:141 cirrhotic patients 124:141 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 5 53 theme permethylated 851:863 arg1 forms 872:876 non-isomeric and isomeric permethylated glycan forms 825:876 non-isomeric and isomeric permethylated glycan forms 825:876 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 12 54 theme glycan 1875:1880 arg1 isoforms 1882:1889 several glycan isoforms 1867:1889 several glycan isoforms 1867:1889 The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis. 28543513 1 55 theme Early 144:148 arg1 detection 156:164 Early stage detection 144:164 Early stage detection 144:164 Early stage detection and cancer treatment demand the identification of reliable biomarkers. 28543513 2 56 theme past 246:249 arg1 decades 251:257 the past decades 242:257 the past decades 242:257 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 5 57 theme forms 872:876 arg1 comparison 811:820 A comparison 809:820 A comparison of non-isomeric and isomeric permethylated glycan forms 809:876 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 3 58 theme haptoglobin 574:584 arg1 glycoprotein 586:597 haptoglobin glycoprotein 574:597 haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 574:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 5 59 theme PGC 930:932 arg1 columns 935:941 porous graphitic carbon (PGC) columns 905:941 porous graphitic carbon (PGC) columns 905:941 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 9 60 theme specific 1529:1536 arg1 structures 1554:1563 the specific isomeric glycan structures 1525:1563 the specific isomeric glycan structures 1525:1563 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 12 61 theme several 1867:1873 arg1 isoforms 1882:1889 several glycan isoforms 1867:1889 several glycan isoforms 1867:1889 The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis. 28543513 9 62 theme glycan 1547:1552 arg1 structures 1554:1563 the specific isomeric glycan structures 1525:1563 the specific isomeric glycan structures 1525:1563 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 11 63 theme isomeric 1778:1785 arg1 separation 1787:1796 isomeric separation 1778:1796 isomeric separation 1778:1796 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 0 64 theme hepatocellular 89:102 arg1 HCC 115:117 HCC 115:117 HCC 115:117 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 0 64 theme hepatocellular 89:102 arg1 carcinoma 104:112 hepatocellular carcinoma 89:112 hepatocellular carcinoma (HCC) 89:118 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 5 65 theme carbon 922:927 arg1 columns 935:941 porous graphitic carbon (PGC) columns 905:941 porous graphitic carbon (PGC) columns 905:941 A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. 28543513 3 66 attach derived 599:605 arg1 sera 612:615 sera 612:615 sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 612:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 3 66 attach derived 599:605 arg2 glycoprotein 586:597 haptoglobin glycoprotein 574:597 haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients 574:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 11 67 theme aberrant 1828:1835 arg1 changes 1844:1850 disease prompted aberrant glycan changes 1811:1850 disease prompted aberrant glycan changes 1811:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 11 68 theme disease 1811:1817 arg1 changes 1844:1850 disease prompted aberrant glycan changes 1811:1850 disease prompted aberrant glycan changes 1811:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 2 69 dep development 417:427 arg1 the 413:415 the 413:415 the 413:415 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 9 70 used utilized 1503:1510 arg2 times 1468:1472 retention times 1458:1472 retention times 1458:1472 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 9 70 used utilized 1503:1510 arg2 spectra 1490:1496 tandem mass spectra 1478:1496 tandem mass spectra 1478:1496 Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. 28543513 10 71 theme linked 1683:1688 arg1 moieties 1702:1709 α-2,6 linked sialic acid moieties 1677:1709 α-2,6 linked sialic acid moieties 1677:1709 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 0 72 theme isomeric 9:16 arg1 profiling 18:26 LC-MS/MS isomeric profiling 0:26 LC-MS/MS isomeric profiling of permethylated N-glycans 0:53 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 8 73 theme isoform 1381:1387 arg1 patterns 1389:1396 The glycan isoform patterns 1370:1396 The glycan isoform patterns 1370:1396 The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. 28543513 2 74 theme proteins 369:376 arg1 variation 300:308 the variation 296:308 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 74 theme proteins 369:376 arg1 structures 355:364 the glycan structures 344:364 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 7 75 theme glycan 1153:1158 arg1 structures 1160:1169 8 out of 34 glycan structures 1141:1169 8 out of 34 glycan structures identified by PGC-LC-MS/MS 1141:1196 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 3 76 theme carcinoma 661:669 arg1 patients 677:684 hepatocellular carcinoma (HCC) patients 646:684 hepatocellular carcinoma (HCC) patients 646:684 Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. 28543513 0 77 theme N-glycans 45:53 arg1 profiling 18:26 LC-MS/MS isomeric profiling 0:26 LC-MS/MS isomeric profiling of permethylated N-glycans 0:53 LC-MS/MS isomeric profiling of permethylated N-glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients. 28543513 1 78 theme reliable 216:223 arg1 biomarkers 225:234 reliable biomarkers 216:234 reliable biomarkers 216:234 Early stage detection and cancer treatment demand the identification of reliable biomarkers. 28543513 6 79 theme sialylated 1045:1054 arg1 structures 1056:1065 10 sialylated structures 1042:1065 10 sialylated structures 1042:1065 In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. 28543513 2 80 theme cancers 459:465 arg1 development 417:427 development 417:427 development 417:427 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 6 81 gly sialylated 1045:1054 arg1 structures 1056:1065 10 sialylated structures 1042:1065 10 sialylated structures 1042:1065 In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. 28543513 8 82 theme stage 1418:1422 arg1 HCC 1424:1426 early stage HCC 1412:1426 early stage HCC 1412:1426 The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. 28543513 10 83 theme acid 1697:1700 arg1 moieties 1702:1709 α-2,6 linked sialic acid moieties 1677:1709 α-2,6 linked sialic acid moieties 1677:1709 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 2 84 from proteins 369:376 arg1 blood 381:385 blood 381:385 blood 381:385 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 84 from proteins 369:376 arg1 serum 390:394 serum 390:394 serum 390:394 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 7 85 theme particular 1286:1295 arg1 composition 1304:1314 a particular glycan composition 1284:1314 a particular glycan composition 1284:1314 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 11 86 theme potential 1754:1762 arg1 importance 1764:1773 the potential importance 1750:1773 the potential importance of isomeric separation for defining disease prompted aberrant glycan changes 1750:1850 The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. 28543513 12 87 theme early 1917:1921 arg1 HCC 1929:1931 early stage HCC 1917:1931 early stage HCC 1917:1931 The levels of several glycan isoforms effectively distinguished early stage HCC from cirrhosis. 28543513 2 88 from blood 381:385 arg1 variation 300:308 the variation 296:308 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 88 from blood 381:385 arg1 structures 355:364 the glycan structures 344:364 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 4 89 theme N-glycans 798:806 arg1 release 787:793 the release 783:793 the release of N-glycans 783:806 The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. 28543513 2 90 theme level 327:331 arg1 variation 300:308 the variation 296:308 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 90 theme level 327:331 arg1 structures 355:364 the glycan structures 344:364 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 91 from serum 390:394 arg1 variation 300:308 the variation 296:308 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 2 91 from serum 390:394 arg1 structures 355:364 the glycan structures 344:364 the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver 296:482 Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. 28543513 7 92 theme isomeric 1258:1265 arg1 distributions 1267:1279 isomeric distributions 1258:1279 isomeric distributions of a particular glycan composition 1258:1314 Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. 28543513 10 93 theme glycan 1577:1582 arg1 significant 1618:1628 significant 1618:1628 significant 1618:1628 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 28543513 10 93 theme glycan 1577:1582 arg1 isomers 1584:1590 the glycan isomers 1573:1590 the glycan isomers 1573:1590 All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. 24108499 8 0 with glucoamylase 1287:1298 arg1 polysaccharides 1305:1319 polysaccharides 1305:1319 polysaccharides 1305:1319 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 2 1 gly O-glycosylated 423:436 arg1 linker 438:443 an O-glycosylated linker 420:443 an O-glycosylated linker 420:443 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 0 2 from structures 8:17 arg1 complex 81:87 complex 81:87 complex with isomaltooligosaccharide 81:116 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 2 from structures 8:17 arg1 glucoamylase 65:76 Rhizopus oryzae glucoamylase 49:76 Rhizopus oryzae glucoamylase 49:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 3 with complex 81:87 arg1 isomaltooligosaccharide 94:116 isomaltooligosaccharide 94:116 isomaltooligosaccharide 94:116 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 5 4 theme site 903:906 arg1 binding 910:916 site I binding 903:916 site I binding 903:916 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 3 5 theme conserved 542:550 arg1 Tyr83 578:582 Tyr83 578:582 Tyr83 578:582 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 5 theme conserved 542:550 arg1 Tyr94 589:593 Tyr94 589:593 Tyr94 589:593 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 5 theme conserved 542:550 arg1 Trp47 571:575 Trp47 571:575 Trp47 571:575 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 5 theme conserved 542:550 arg1 residues 561:568 three highly conserved aromatic residues 529:568 three highly conserved aromatic residues 529:568 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 8 6 from sites 1233:1237 arg1 RoSBD 1242:1246 RoSBD 1242:1246 RoSBD 1242:1246 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 3 7 theme aromatic 552:559 arg1 Tyr83 578:582 Tyr83 578:582 Tyr83 578:582 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 7 theme aromatic 552:559 arg1 Tyr94 589:593 Tyr94 589:593 Tyr94 589:593 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 7 theme aromatic 552:559 arg1 Trp47 571:575 Trp47 571:575 Trp47 571:575 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 3 7 theme aromatic 552:559 arg1 residues 561:568 three highly conserved aromatic residues 529:568 three highly conserved aromatic residues 529:568 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 5 8 theme site 850:853 arg1 binding 858:864 site II binding 850:864 site II binding 850:864 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 8 9 theme binding 1225:1231 arg1 sites 1233:1237 two carbohydrate binding sites 1208:1237 two carbohydrate binding sites in RoSBD 1208:1246 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 3 10 theme carbohydrate 450:461 arg1 sites 471:475 Two carbohydrate binding sites 446:475 Two carbohydrate binding sites in RoSBD 446:484 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 8 11 theme carbohydrate 1212:1223 arg1 sites 1233:1237 two carbohydrate binding sites 1208:1237 two carbohydrate binding sites in RoSBD 1208:1246 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 3 12 theme binding 463:469 arg1 sites 471:475 Two carbohydrate binding sites 446:475 Two carbohydrate binding sites in RoSBD 446:484 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 5 13 theme I 908:908 arg1 binding 910:916 site I binding 903:916 site I binding 903:916 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 2 14 theme N-terminal 334:343 arg1 domain 360:365 an N-terminal starch binding domain 331:365 an N-terminal starch binding domain (SBD) 331:371 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 2 14 theme N-terminal 334:343 arg1 SBD 368:370 SBD 368:370 SBD 368:370 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 7 15 theme glucosyl 1137:1144 arg1 units 1146:1150 glucosyl units 1137:1150 glucosyl units of oligosaccharides 1137:1170 Site I participates in sugar binding only when the number of glucosyl units of oligosaccharides is more than three. 24108499 2 16 theme catalytic 390:398 arg1 domain 400:405 a C-terminal catalytic domain 377:405 a C-terminal catalytic domain connected by an O-glycosylated linker 377:443 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 0 17 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 0:181 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 2 18 theme C-terminal 379:388 arg1 domain 400:405 a C-terminal catalytic domain 377:405 a C-terminal catalytic domain connected by an O-glycosylated linker 377:443 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 0 19 theme starch 22:27 arg1 domain 37:42 starch binding domain 22:42 starch binding domain from Rhizopus oryzae glucoamylase 22:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 2 20 theme Rhizopus 280:287 arg1 oryzae 289:294 Rhizopus oryzae glucoamylase (RoGA) 280:314 Rhizopus oryzae glucoamylase (RoGA) 280:314 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 0 21 theme polysaccharide 133:146 arg1 mechanism 156:164 polysaccharide binding mechanism 133:164 polysaccharide binding mechanism of CBM21 family 133:180 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 6 22 theme binding 999:1005 arg1 site 1007:1010 the recognition binding site 983:1010 the recognition binding site for carbohydrate 983:1027 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 6 22 theme binding 999:1005 arg1 site 967:970 site II 967:973 site II 967:973 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 6 22 theme binding 999:1005 arg1 site 1053:1056 site II 1053:1059 site II 1053:1059 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 2 23 theme O-glycosylated 423:436 arg1 linker 438:443 an O-glycosylated linker 420:443 an O-glycosylated linker 420:443 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 0 24 theme domain 37:42 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 0:181 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 7 25 theme oligosaccharides 1155:1170 arg1 units 1146:1150 glucosyl units 1137:1150 glucosyl units of oligosaccharides 1137:1170 Site I participates in sugar binding only when the number of glucosyl units of oligosaccharides is more than three. 24108499 2 26 dep oryzae 289:294 arg1 RoGA 310:313 RoGA 310:313 RoGA 310:313 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 2 26 dep oryzae 289:294 arg1 glucoamylase 296:307 glucoamylase 296:307 Rhizopus oryzae glucoamylase (RoGA) 280:314 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 1 27 theme starch 231:236 arg1 hydrolysis 217:226 hydrolysis 217:226 hydrolysis of starch and polysaccharides to yield β-D-glucose 217:277 Glucoamylases are responsible for hydrolysis of starch and polysaccharides to yield β-D-glucose. 24108499 0 28 theme binding 29:35 arg1 domain 37:42 starch binding domain 22:42 starch binding domain from Rhizopus oryzae glucoamylase 22:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 29 theme binding 148:154 arg1 mechanism 156:164 polysaccharide binding mechanism 133:164 polysaccharide binding mechanism of CBM21 family 133:180 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 30 from glucoamylase 65:76 arg1 domain 37:42 starch binding domain 22:42 starch binding domain from Rhizopus oryzae glucoamylase 22:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 30 from glucoamylase 65:76 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 0:181 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 3 31 from sites 471:475 arg1 RoSBD 480:484 RoSBD 480:484 RoSBD 480:484 Two carbohydrate binding sites in RoSBD have been identified, site I is created by three highly conserved aromatic residues, Trp47, Tyr83, and Tyr94, and site II is built up by Tyr32 and Phe58. 24108499 4 32 link -linked 710:716 arg1 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 32 link -linked 710:716 arg1 1,6 706:708 1,6 706:708 1,6 706:708 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 32 link -linked 710:716 arg1 isomaltotriose 718:731 only α-(1,6)-linked isomaltotriose 698:731 only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) 698:745 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 0 33 theme Rhizopus 49:56 arg1 oryzae 58:63 Rhizopus oryzae 49:63 Rhizopus oryzae glucoamylase 49:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 0 34 theme CBM21 169:173 arg1 family 175:180 CBM21 family 169:180 CBM21 family 169:180 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 7 35 theme units 1146:1150 arg1 more 1175:1178 more 1175:1178 more 1175:1178 Site I participates in sugar binding only when the number of glucosyl units of oligosaccharides is more than three. 24108499 7 35 theme units 1146:1150 arg1 number 1127:1132 the number 1123:1132 the number of glucosyl units of oligosaccharides 1123:1170 Site I participates in sugar binding only when the number of glucosyl units of oligosaccharides is more than three. 24108499 4 36 with complex 685:691 arg1 1,6 706:708 1,6 706:708 1,6 706:708 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 36 with complex 685:691 arg1 RoSBD-isoG4 769:779 RoSBD-isoG4 769:779 RoSBD-isoG4 769:779 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 36 with complex 685:691 arg1 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 36 with complex 685:691 arg1 isomaltotetraose 751:766 isomaltotetraose 751:766 isomaltotetraose (RoSBD-isoG4) 751:780 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 36 with complex 685:691 arg1 isomaltotriose 718:731 only α-(1,6)-linked isomaltotriose 698:731 only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) 698:745 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 8 37 theme binding 1271:1277 arg1 mode 1279:1282 binding mode 1271:1282 binding mode of glucoamylase with polysaccharides as well as the starch 1271:1341 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 4 38 theme crystal 654:660 arg1 structures 662:671 the two crystal structures 646:671 the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) 646:780 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 5 39 theme RoSBD-isoG4 939:949 arg1 complex 951:957 the RoSBD-isoG4 complex 935:957 the RoSBD-isoG4 complex 935:957 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 8 40 theme glucoamylase 1287:1298 arg1 mode 1279:1282 binding mode 1271:1282 binding mode of glucoamylase with polysaccharides as well as the starch 1271:1341 Taken together, two carbohydrate binding sites in RoSBD cooperate to reinforce binding mode of glucoamylase with polysaccharides as well as the starch. 24108499 2 41 theme binding 352:358 arg1 domain 360:365 an N-terminal starch binding domain 331:365 an N-terminal starch binding domain (SBD) 331:371 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 2 41 theme binding 352:358 arg1 SBD 368:370 SBD 368:370 SBD 368:370 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 1 42 theme polysaccharides 242:256 arg1 hydrolysis 217:226 hydrolysis 217:226 hydrolysis of starch and polysaccharides to yield β-D-glucose 217:277 Glucoamylases are responsible for hydrolysis of starch and polysaccharides to yield β-D-glucose. 24108499 0 43 theme family 175:180 arg1 mechanism 156:164 polysaccharide binding mechanism 133:164 polysaccharide binding mechanism of CBM21 family 133:180 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 4 44 theme -linked 710:716 arg1 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 RoSBD-isoG3 734:744 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 44 theme -linked 710:716 arg1 1,6 706:708 1,6 706:708 1,6 706:708 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 44 theme -linked 710:716 arg1 isomaltotriose 718:731 only α-(1,6)-linked isomaltotriose 698:731 only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) 698:745 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 2 45 theme starch 345:350 arg1 domain 360:365 an N-terminal starch binding domain 331:365 an N-terminal starch binding domain (SBD) 331:371 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 2 45 theme starch 345:350 arg1 SBD 368:370 SBD 368:370 SBD 368:370 Rhizopus oryzae glucoamylase (RoGA) is composed of an N-terminal starch binding domain (SBD) and a C-terminal catalytic domain connected by an O-glycosylated linker. 24108499 4 46 from structures 662:671 arg1 complex 685:691 complex 685:691 complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) 685:780 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 4 47 theme RoSBD 676:680 arg1 structures 662:671 the two crystal structures 646:671 the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) 646:780 Here, the two crystal structures of RoSBD in complex with only α-(1,6)-linked isomaltotriose (RoSBD-isoG3) and isomaltotetraose (RoSBD-isoG4) have been determined at 1.2 and 1.3 Å, respectively. 24108499 7 48 theme sugar 1099:1103 arg1 binding 1105:1111 sugar binding 1099:1111 sugar binding 1099:1111 Site I participates in sugar binding only when the number of glucosyl units of oligosaccharides is more than three. 24108499 0 49 dep structures 8:17 arg1 insights 119:126 insights 119:126 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 0:181 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 6 50 theme recognition 987:997 arg1 site 1007:1010 the recognition binding site 983:1010 the recognition binding site for carbohydrate 983:1027 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 6 50 theme recognition 987:997 arg1 site 967:970 site II 967:973 site II 967:973 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 6 50 theme recognition 987:997 arg1 site 1053:1056 site II 1053:1059 site II 1053:1059 Hence, site II acts as the recognition binding site for carbohydrate and site I accommodates site II to bind isoG4. 24108499 0 51 theme oryzae 58:63 arg1 glucoamylase 65:76 Rhizopus oryzae glucoamylase 49:76 Rhizopus oryzae glucoamylase 49:76 Crystal structures of starch binding domain from Rhizopus oryzae glucoamylase in complex with isomaltooligosaccharide: insights into polysaccharide binding mechanism of CBM21 family. 24108499 5 52 located observed 869:876 arg1 complexes 886:894 both complexes 881:894 both complexes 881:894 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 5 52 located observed 869:876 arg2 binding 858:864 site II binding 850:864 site II binding 850:864 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 5 53 located found 926:930 arg1 complex 951:957 the RoSBD-isoG4 complex 935:957 the RoSBD-isoG4 complex 935:957 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24108499 5 53 located found 926:930 arg2 binding 910:916 site I binding 903:916 site I binding 903:916 Interestingly, site II binding is observed in both complexes, while site I binding is only found in the RoSBD-isoG4 complex. 24970053 0 0 theme upstream 66:73 arg1 role 26:29 an important role 13:29 an important role 13:29 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 0 0 theme upstream 66:73 arg1 modulator 75:83 an upstream modulator 63:83 an upstream modulator of MMP production in tumor cells 63:116 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 5 1 theme cancer 773:778 arg1 MCF-7 792:796 MCF-7 792:796 MCF-7 792:796 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 U87 812:814 U87 812:814 U87 812:814 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 SGC-7901 817:824 SGC-7901 817:824 SGC-7901 817:824 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 M231 799:802 M231 799:802 M231 799:802 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 LN229 805:809 LN229 805:809 LN229 805:809 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 U251 830:833 U251 830:833 U251 830:833 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 1 theme cancer 773:778 arg1 lines 785:789 6 cancer cell lines 771:789 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 771:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 8 2 used used 1161:1164 arg2 tunicamycin 1139:1149 tunicamycin 1139:1149 tunicamycin 1139:1149 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 8 2 used used 1161:1164 arg2 inhibitor 1172:1180 an inhibitor 1169:1180 an inhibitor of N-linked glycosylation 1169:1206 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 6 3 theme other 940:944 arg1 lines 951:955 the other cell lines 936:955 the other cell lines 936:955 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 5 4 theme cell 780:783 arg1 MCF-7 792:796 MCF-7 792:796 MCF-7 792:796 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 U87 812:814 U87 812:814 U87 812:814 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 SGC-7901 817:824 SGC-7901 817:824 SGC-7901 817:824 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 M231 799:802 M231 799:802 M231 799:802 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 LN229 805:809 LN229 805:809 LN229 805:809 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 U251 830:833 U251 830:833 U251 830:833 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 4 theme cell 780:783 arg1 lines 785:789 6 cancer cell lines 771:789 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 771:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 4 5 theme major 547:551 arg1 carrier 553:559 a major carrier 545:559 a major carrier of β1-6-branched polylactosamine sugars on tumor cells 545:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 5 theme major 547:551 arg1 CD147 536:540 CD147 536:540 CD147 536:540 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 9 6 from influence 1305:1313 arg1 N-glycosylation 1324:1338 CD147 N-glycosylation 1318:1338 CD147 N-glycosylation 1318:1338 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 9 6 from influence 1305:1313 arg1 expression 1350:1359 MMP-2 expression 1344:1359 MMP-2 expression 1344:1359 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 4 7 gly glycoform 630:638 arg1 HG-CD147 650:657 HG-CD147 650:657 HG-CD147 650:657 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 7 gly glycoform 630:638 arg1 CD147 643:647 CD147 643:647 CD147 (HG-CD147) 643:658 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 1 8 theme related 146:152 arg1 glycosyl-transferases 154:174 related glycosyl-transferases 146:174 related glycosyl-transferases 146:174 Aberrant carbohydration by related glycosyl-transferases plays an important role in the progression of cancer. 24970053 10 9 theme important 1481:1489 arg1 effect 1491:1496 an important effect 1478:1496 an important effect 1478:1496 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 5 10 from expression 757:766 arg1 MCF-7 792:796 MCF-7 792:796 MCF-7 792:796 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 U87 812:814 U87 812:814 U87 812:814 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 SGC-7901 817:824 SGC-7901 817:824 SGC-7901 817:824 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 M231 799:802 M231 799:802 M231 799:802 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 LN229 805:809 LN229 805:809 LN229 805:809 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 U251 830:833 U251 830:833 U251 830:833 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 10 from expression 757:766 arg1 lines 785:789 6 cancer cell lines 771:789 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 771:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 9 11 theme tunicamycin 1242:1252 arg1 concentrations 1224:1237 various concentrations 1216:1237 various concentrations of tunicamycin 1216:1252 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 7 12 theme cell 1048:1051 arg1 lines 1053:1057 the LN229 and SGC-7901 cell lines 1025:1057 lines 1053:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 10 13 theme MMP-2 1501:1505 arg1 expression 1507:1516 MMP-2 expression 1501:1516 MMP-2 expression 1501:1516 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 4 14 theme high 625:628 arg1 glycoform 630:638 the high glycoform 621:638 the high glycoform of CD147 (HG-CD147) 621:658 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 5 15 theme β3GnT8 745:750 arg1 expression 757:766 β3GnT8 mRNA expression 745:766 β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 745:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 2 16 from regulation 353:362 arg1 cells 415:419 cancer cells 408:419 cancer cells 408:419 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 2 17 theme β-1,3-N-acetyl-glucosaminyltransferase-8 266:305 arg1 ablity 256:261 the ablity 252:261 the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) 252:314 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 3 18 theme tetraantennary 510:523 arg1 N-glycans 525:533 β1-6-branched tetraantennary N-glycans 496:533 β1-6-branched tetraantennary N-glycans 496:533 β3GnT8 catalyzes and then extends a polylactosamine chain specifically on β1-6-branched tetraantennary N-glycans. 24970053 0 19 theme production 92:101 arg1 role 26:29 an important role 13:29 an important role 13:29 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 0 19 theme production 92:101 arg1 modulator 75:83 an upstream modulator 63:83 an upstream modulator of MMP production in tumor cells 63:116 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 6 20 theme β3GnT8 851:856 arg1 expression 858:867 β3GnT8 expression 851:867 β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines 851:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 7 21 theme CD147 1103:1107 arg1 level 1074:1078 the level 1070:1078 the level of polylactosamine and CD147 N-glycosylation 1070:1123 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 8 22 theme glycosylation 1194:1206 arg1 tunicamycin 1139:1149 tunicamycin 1139:1149 tunicamycin 1139:1149 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 8 22 theme glycosylation 1194:1206 arg1 inhibitor 1172:1180 an inhibitor 1169:1180 an inhibitor of N-linked glycosylation 1169:1206 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 10 23 contain has 1474:1476 arg2 effect 1491:1496 an important effect 1478:1496 an important effect 1478:1496 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 10 23 contain has 1474:1476 arg1 N-glycosylation 1449:1463 N-glycosylation 1449:1463 N-glycosylation of CD147 1449:1472 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 8 24 theme N-linked 1185:1192 arg1 glycosylation 1194:1206 N-linked glycosylation 1185:1206 N-linked glycosylation 1185:1206 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 2 25 theme cancer 408:413 arg1 cells 415:419 cancer cells 408:419 cancer cells 408:419 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 0 26 theme MMP 88:90 arg1 production 92:101 MMP production 88:101 MMP production 88:101 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 7 27 theme SGC-7901 1039:1046 arg1 lines 1053:1057 the LN229 and SGC-7901 cell lines 1025:1057 lines 1053:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 9 28 theme CD147 1318:1322 arg1 N-glycosylation 1324:1338 CD147 N-glycosylation 1318:1338 CD147 N-glycosylation 1318:1338 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 4 29 theme metalloproteinase 675:691 arg1 production 699:708 matrix metalloproteinase (MMP) production 668:708 matrix metalloproteinase (MMP) production 668:708 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 30 from cells 610:614 arg1 carrier 553:559 a major carrier 545:559 a major carrier of β1-6-branched polylactosamine sugars on tumor cells 545:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 30 from cells 610:614 arg1 CD147 536:540 CD147 536:540 CD147 536:540 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 11 31 theme MMP-2 1595:1599 arg1 pathway 1584:1590 the signal transduction pathway 1560:1590 the signal transduction pathway of MMP-2 1560:1599 Our findings suggest that β3GnT8 affects the signal transduction pathway of MMP-2 by altering the N-glycan structure of CD147. 24970053 5 32 dep lines 785:789 arg1 MCF-7 792:796 MCF-7 792:796 MCF-7 792:796 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 U87 812:814 U87 812:814 U87 812:814 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 SGC-7901 817:824 SGC-7901 817:824 SGC-7901 817:824 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 M231 799:802 M231 799:802 M231 799:802 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 LN229 805:809 LN229 805:809 LN229 805:809 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 U251 830:833 U251 830:833 U251 830:833 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 5 32 dep lines 785:789 arg1 lines 785:789 6 cancer cell lines 771:789 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 771:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 7 33 theme β3GnT8-knockdown 984:999 arg1 lines 1006:1010 β3GnT8-knockdown cell lines 984:1010 β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines 984:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 2 34 theme pathway 397:403 arg1 regulation 353:362 regulation 353:362 regulation of the CD147 signal transduction pathway in cancer cells 353:419 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 7 35 dep polylactosamine 1083:1097 arg1 N-glycosylation 1109:1123 N-glycosylation 1109:1123 N-glycosylation 1109:1123 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 10 36 gly N-glycosylation 1449:1463 arg1 CD147 1468:1472 CD147 1468:1472 CD147 1468:1472 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 2 37 theme transduction 384:395 arg1 pathway 397:403 the CD147 signal transduction pathway 367:403 the CD147 signal transduction pathway 367:403 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 0 38 theme important 16:24 arg1 modulator 75:83 an upstream modulator 63:83 an upstream modulator of MMP production in tumor cells 63:116 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 0 38 theme important 16:24 arg1 role 26:29 an important role 13:29 an important role 13:29 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 4 39 theme sugars 594:599 arg1 carrier 553:559 a major carrier 545:559 a major carrier of β1-6-branched polylactosamine sugars on tumor cells 545:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 39 theme sugars 594:599 arg1 CD147 536:540 CD147 536:540 CD147 536:540 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 0 40 theme tumor 106:110 arg1 cells 112:116 tumor cells 106:116 tumor cells 106:116 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 7 41 attach derived 1012:1018 arg1 lines 1053:1057 the LN229 and SGC-7901 cell lines 1025:1057 lines 1053:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 7 41 attach derived 1012:1018 arg2 lines 1006:1010 β3GnT8-knockdown cell lines 984:1010 β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines 984:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 7 41 attach derived 1012:1018 arg1 LN229 1029:1033 the LN229 and SGC-7901 cell lines 1025:1057 LN229 1029:1033 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 4 42 from sugars 594:599 arg1 cells 610:614 tumor cells 604:614 tumor cells 604:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 10 43 theme N-glycans 1421:1429 arg1 level 1412:1416 the level 1408:1416 the level of N-glycans on CD147 1408:1438 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 8 44 link N-linked 1185:1192 arg1 glycosylation 1194:1206 N-linked glycosylation 1185:1206 N-linked glycosylation 1185:1206 In addition, tunicamycin is widely used as an inhibitor of N-linked glycosylation. 24970053 6 45 theme LN229 876:880 arg1 lines 906:910 the LN229, SGC-7901 and U251 cell lines 872:910 lines 906:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 1 46 theme Aberrant 119:126 arg1 carbohydration 128:141 Aberrant carbohydration 119:141 Aberrant carbohydration by related glycosyl-transferases 119:174 Aberrant carbohydration by related glycosyl-transferases plays an important role in the progression of cancer. 24970053 4 47 theme tumor 604:608 arg1 cells 610:614 tumor cells 604:614 tumor cells 604:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 7 48 theme cell 1001:1004 arg1 lines 1006:1010 β3GnT8-knockdown cell lines 984:1010 β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines 984:1057 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 4 49 theme matrix 668:673 arg1 MMP 694:696 MMP 694:696 MMP 694:696 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 4 49 theme matrix 668:673 arg1 metalloproteinase 675:691 matrix metalloproteinase 668:691 matrix metalloproteinase (MMP) production 668:708 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 1 50 theme important 185:193 arg1 role 195:198 an important role 182:198 an important role 182:198 Aberrant carbohydration by related glycosyl-transferases plays an important role in the progression of cancer. 24970053 3 51 theme polylactosamine 458:472 arg1 chain 474:478 a polylactosamine chain 456:478 a polylactosamine chain 456:478 β3GnT8 catalyzes and then extends a polylactosamine chain specifically on β1-6-branched tetraantennary N-glycans. 24970053 7 52 theme polylactosamine 1083:1097 arg1 level 1074:1078 the level 1070:1078 the level of polylactosamine and CD147 N-glycosylation 1070:1123 Therefore, we established β3GnT8-knockdown cell lines derived from the LN229 and SGC-7901 cell lines to examine the level of polylactosamine and CD147 N-glycosylation. 24970053 6 53 theme cell 901:904 arg1 lines 906:910 the LN229, SGC-7901 and U251 cell lines 872:910 lines 906:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 11 54 theme signal 1564:1569 arg1 pathway 1584:1590 the signal transduction pathway 1560:1590 the signal transduction pathway of MMP-2 1560:1599 Our findings suggest that β3GnT8 affects the signal transduction pathway of MMP-2 by altering the N-glycan structure of CD147. 24970053 9 55 theme MMP-2 1344:1348 arg1 expression 1350:1359 MMP-2 expression 1344:1359 MMP-2 expression 1344:1359 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 6 56 theme SGC-7901 883:890 arg1 lines 906:910 the LN229, SGC-7901 and U251 cell lines 872:910 lines 906:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 0 57 theme signal 40:45 arg1 transduction 47:58 CD147 signal transduction 34:58 CD147 signal transduction 34:58 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 6 58 theme U251 896:899 arg1 lines 906:910 the LN229, SGC-7901 and U251 cell lines 872:910 lines 906:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 4 59 from carrier 553:559 arg1 cells 610:614 tumor cells 604:614 tumor cells 604:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 11 60 theme N-glycan 1617:1624 arg1 structure 1626:1634 the N-glycan structure 1613:1634 the N-glycan structure of CD147 1613:1643 Our findings suggest that β3GnT8 affects the signal transduction pathway of MMP-2 by altering the N-glycan structure of CD147. 24970053 0 61 theme CD147 34:38 arg1 transduction 47:58 CD147 signal transduction 34:58 CD147 signal transduction 34:58 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 4 62 theme polylactosamine 578:592 arg1 sugars 594:599 β1-6-branched polylactosamine sugars 564:599 β1-6-branched polylactosamine sugars on tumor cells 564:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 5 63 theme present 718:724 arg1 study 726:730 the present study 714:730 the present study 714:730 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 10 64 theme CD147 1468:1472 arg1 N-glycosylation 1449:1463 N-glycosylation 1449:1463 N-glycosylation of CD147 1449:1472 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 9 65 used used 1259:1262 arg2 concentrations 1224:1237 various concentrations 1216:1237 various concentrations of tunicamycin 1216:1252 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 4 66 theme β1-6-branched 564:576 arg1 sugars 594:599 β1-6-branched polylactosamine sugars 564:599 β1-6-branched polylactosamine sugars on tumor cells 564:614 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 24970053 6 67 from expression 858:867 arg1 lines 906:910 the LN229, SGC-7901 and U251 cell lines 872:910 lines 906:910 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 3 68 theme β1-6-branched 496:508 arg1 N-glycans 525:533 β1-6-branched tetraantennary N-glycans 496:533 β1-6-branched tetraantennary N-glycans 496:533 β3GnT8 catalyzes and then extends a polylactosamine chain specifically on β1-6-branched tetraantennary N-glycans. 24970053 5 69 theme mRNA 752:755 arg1 expression 757:766 β3GnT8 mRNA expression 745:766 β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251) 745:834 In the present study, we analyzed β3GnT8 mRNA expression in 6 cancer cell lines (MCF-7, M231, LN229, U87, SGC-7901 and U251). 24970053 11 70 theme CD147 1639:1643 arg1 structure 1626:1634 the N-glycan structure 1613:1634 the N-glycan structure of CD147 1613:1643 Our findings suggest that β3GnT8 affects the signal transduction pathway of MMP-2 by altering the N-glycan structure of CD147. 24970053 2 71 theme MMP-2 328:332 arg1 expression 334:343 MMP-2 expression 328:343 MMP-2 expression 328:343 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 1 72 theme cancer 222:227 arg1 progression 207:217 the progression 203:217 the progression of cancer 203:227 Aberrant carbohydration by related glycosyl-transferases plays an important role in the progression of cancer. 24970053 0 73 from modulator 75:83 arg1 cells 112:116 tumor cells 106:116 tumor cells 106:116 β3GnT8 plays an important role in CD147 signal transduction as an upstream modulator of MMP production in tumor cells. 24970053 2 74 theme signal 377:382 arg1 pathway 397:403 the CD147 signal transduction pathway 367:403 the CD147 signal transduction pathway 367:403 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 10 75 from level 1412:1416 arg1 CD147 1434:1438 CD147 1434:1438 CD147 1434:1438 In conclusion, we found that β3GnT8 regulated the level of N-glycans on CD147 and that N-glycosylation of CD147 has an important effect on MMP-2 expression. 24970053 11 76 theme transduction 1571:1582 arg1 pathway 1584:1590 the signal transduction pathway 1560:1590 the signal transduction pathway of MMP-2 1560:1599 Our findings suggest that β3GnT8 affects the signal transduction pathway of MMP-2 by altering the N-glycan structure of CD147. 24970053 9 77 theme various 1216:1222 arg1 concentrations 1224:1237 various concentrations 1216:1237 various concentrations of tunicamycin 1216:1252 Hence, various concentrations of tunicamycin were used to treat the cells in order to study its influence on CD147 N-glycosylation and MMP-2 expression. 24970053 6 78 theme cell 946:949 arg1 lines 951:955 the other cell lines 936:955 the other cell lines 936:955 We found that β3GnT8 expression in the LN229, SGC-7901 and U251 cell lines was higher than that in the other cell lines. 24970053 2 79 theme CD147 371:375 arg1 pathway 397:403 the CD147 signal transduction pathway 367:403 the CD147 signal transduction pathway 367:403 This study focused on the ablity of β-1,3-N-acetyl-glucosaminyltransferase-8 (β3GnT8) to regulate MMP-2 expression through regulation of the CD147 signal transduction pathway in cancer cells. 24970053 4 80 theme CD147 643:647 arg1 glycoform 630:638 the high glycoform 621:638 the high glycoform of CD147 (HG-CD147) 621:658 CD147 is a major carrier of β1-6-branched polylactosamine sugars on tumor cells, and the high glycoform of CD147 (HG-CD147) induces matrix metalloproteinase (MMP) production. 26544759 4 0 theme branched 1150:1157 arg1 sugars 1159:1164 the linear and branched sugars 1135:1164 the linear and branched sugars 1135:1164 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 1 theme molecular 391:399 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 1 theme molecular 391:399 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 1 theme molecular 391:399 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 1 theme molecular 391:399 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 2 theme different 955:963 arg1 molecules 971:979 the different sugar molecules 951:979 the different sugar molecules 951:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 1 3 theme fucosylated 278:288 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 0 4 theme Capillary 75:83 arg1 Electrophoresis 85:99 Capillary Electrophoresis 75:99 Capillary Electrophoresis 75:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 2 5 theme high 386:389 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 5 theme high 386:389 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 5 theme high 386:389 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 5 theme high 386:389 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 1 6 theme biantennary 290:300 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 4 7 theme energy 993:998 arg1 values 1000:1005 Activation energy values 982:1005 Activation energy values 982:1005 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 0 8 from Effect 0:5 arg1 Migration 62:70 Structure Specific Glycan Migration 36:70 Structure Specific Glycan Migration in Capillary Electrophoresis 36:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 2 9 theme Background 330:339 arg1 electrolytes 341:352 Background electrolytes 330:352 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 330:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 4 10 theme linear 1139:1144 arg1 sugars 1159:1164 the linear and branched sugars 1135:1164 the linear and branched sugars 1135:1164 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 1 11 theme IgG 302:304 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 4 12 theme various 1185:1191 arg1 additives 1216:1224 the various background electrolyte additives 1181:1224 the various background electrolyte additives 1181:1224 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 4 13 theme electrolyte 1204:1214 arg1 additives 1216:1224 the various background electrolyte additives 1181:1224 the various background electrolyte additives 1181:1224 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 3 14 theme molecules 971:979 arg1 migration 938:946 the structure specific electrophoretic migration 899:946 the structure specific electrophoretic migration of the different sugar molecules 899:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 2 15 theme IgG 644:646 arg1 glycans 648:654 the all IgG glycans 636:654 the all IgG glycans 636:654 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 16 with electrolytes 341:352 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 16 with electrolytes 341:352 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 16 with electrolytes 341:352 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 16 with electrolytes 341:352 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 17 theme glycosylation 790:802 arg1 analysis 804:811 glycosylation analysis 790:811 glycosylation analysis by capillary electrophoresis 790:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 18 theme temperature 528:538 arg1 range 540:544 the temperature range 524:544 the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 524:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 4 19 theme mobility 1074:1081 arg1 plots 1053:1057 the Arrhenius plots 1039:1057 the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature 1039:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 3 20 theme sugar 965:969 arg1 molecules 971:979 the different sugar molecules 951:979 the different sugar molecules 951:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 4 21 theme temperature 1106:1116 arg1 plots 1053:1057 the Arrhenius plots 1039:1057 the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature 1039:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 22 dep value 577:581 arg1 shifts 583:588 shifts 583:588 shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 583:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 1 23 gly fucosylated 278:288 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 2 24 theme control 775:781 arg1 importance 743:752 the importance 739:752 the importance of tight temperature control 739:781 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 25 theme ethylene 419:426 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 25 theme ethylene 419:426 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 4 26 theme absolute 1097:1104 arg1 temperature 1106:1116 reciprocal absolute temperature 1086:1116 reciprocal absolute temperature 1086:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 0 27 theme Temperature 21:31 arg1 Effect 0:5 Effect 0:5 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 0:100 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 1 28 theme dependent 114:122 arg1 shifts 147:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 0 29 theme Separation 10:19 arg1 Temperature 21:31 Separation Temperature 10:31 Separation Temperature 10:31 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 2 30 theme unit 567:570 arg1 value 577:581 20-50 °C. Glucose unit (GU) value 549:581 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 549:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 1 31 theme differential 124:135 arg1 shifts 147:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 2 32 theme capillary 816:824 arg1 electrophoresis 826:840 capillary electrophoresis 816:840 capillary electrophoresis 816:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 0 33 theme Structure 36:44 arg1 Migration 62:70 Structure Specific Glycan Migration 36:70 Structure Specific Glycan Migration in Capillary Electrophoresis 36:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 4 34 theme background 1193:1202 arg1 additives 1216:1224 the various background electrolyte additives 1181:1224 the various background electrolyte additives 1181:1224 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 35 theme Glucose 559:565 arg1 value 577:581 20-50 °C. Glucose unit (GU) value 549:581 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 549:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 36 dep additives 408:416 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 36 dep additives 408:416 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 36 dep additives 408:416 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 36 dep additives 408:416 arg1 oxide 476:480 ethylene oxide 467:480 ethylene oxide 467:480 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 36 dep additives 408:416 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 4 37 theme logarithmic 1062:1072 arg1 mobility 1074:1081 logarithmic mobility 1062:1081 logarithmic mobility 1062:1081 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 4 38 theme plots 1053:1057 arg1 slopes 1029:1034 the slopes 1025:1034 the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature 1025:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 39 theme °C. 555:557 arg1 value 577:581 20-50 °C. Glucose unit (GU) value 549:581 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 549:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 40 theme ethylene 467:474 arg1 oxide 476:480 ethylene oxide 467:480 ethylene oxide 467:480 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 40 theme ethylene 467:474 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 41 theme linear 436:441 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 41 theme linear 436:441 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 0 42 theme Glycan 55:60 arg1 Migration 62:70 Structure Specific Glycan Migration 36:70 Structure Specific Glycan Migration in Capillary Electrophoresis 36:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 0 43 from Migration 62:70 arg1 Electrophoresis 85:99 Capillary Electrophoresis 75:99 Capillary Electrophoresis 75:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 1 44 dep linear 204:209 arg1 carbohydrates 315:327 carbohydrates 315:327 carbohydrates 315:327 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 1 45 theme migration 137:145 arg1 shifts 147:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts 102:152 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 0 46 theme Specific 46:53 arg1 Migration 62:70 Structure Specific Glycan Migration 36:70 Structure Specific Glycan Migration in Capillary Electrophoresis 36:99 Effect of Separation Temperature on Structure Specific Glycan Migration in Capillary Electrophoresis. 26544759 2 47 theme value 577:581 arg1 range 540:544 the temperature range 524:544 the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 524:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 48 theme activation 847:856 arg1 concept 865:871 The activation energy concept 843:871 The activation energy concept 843:871 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 2 49 theme low 378:380 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 49 theme low 378:380 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 49 theme low 378:380 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 49 theme low 378:380 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 50 theme specific 913:920 arg1 migration 938:946 the structure specific electrophoretic migration 899:946 the structure specific electrophoretic migration of the different sugar molecules 899:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 4 51 theme Activation 982:991 arg1 values 1000:1005 Activation energy values 982:1005 Activation energy values 982:1005 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 4 52 attach derived 1012:1018 arg1 slopes 1029:1034 the slopes 1025:1034 the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature 1025:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 4 52 attach derived 1012:1018 arg2 values 1000:1005 Activation energy values 982:1005 Activation energy values 982:1005 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 53 dep additive-free 664:676 arg1 electrolytes 713:724 electrolytes 713:724 electrolytes 713:724 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 54 theme energy 858:863 arg1 concept 865:871 The activation energy concept 843:871 The activation energy concept 843:871 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 2 55 theme GU 573:574 arg1 value 577:581 20-50 °C. Glucose unit (GU) value 549:581 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis 549:840 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 56 theme temperature 763:773 arg1 control 775:781 tight temperature control 757:781 tight temperature control 757:781 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 57 theme weight 401:406 arg1 glycol 428:433 ethylene glycol 419:433 ethylene glycol 419:433 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 57 theme weight 401:406 arg1 polyacrylamide 443:456 linear polyacrylamide 436:456 linear polyacrylamide 436:456 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 57 theme weight 401:406 arg1 poly 462:465 poly 462:465 poly(ethylene oxide) 462:481 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 2 57 theme weight 401:406 arg1 additives 408:416 low and high molecular weight additives 378:416 low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) 378:482 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 58 theme electrophoretic 922:936 arg1 migration 938:946 the structure specific electrophoretic migration 899:946 the structure specific electrophoretic migration of the different sugar molecules 899:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 2 59 theme additive-containing 682:700 arg1 background 702:711 additive-containing background 682:711 additive-containing background 682:711 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 4 60 theme Arrhenius 1043:1051 arg1 plots 1053:1057 the Arrhenius plots 1039:1057 the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature 1039:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 2 61 theme tight 757:761 arg1 control 775:781 tight temperature control 757:781 tight temperature control 757:781 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 3 62 theme structure 903:911 arg1 migration 938:946 the structure specific electrophoretic migration 899:946 the structure specific electrophoretic migration of the different sugar molecules 899:979 The activation energy concept was applied to understand the structure specific electrophoretic migration of the different sugar molecules. 26544759 4 63 theme reciprocal 1086:1095 arg1 temperature 1106:1116 reciprocal absolute temperature 1086:1116 reciprocal absolute temperature 1086:1116 Activation energy values were derived from the slopes of the Arrhenius plots of logarithmic mobility vs reciprocal absolute temperature and compared for the linear and branched sugars as well as for the various background electrolyte additives. 26544759 1 64 dep branched 239:246 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 1 64 dep branched 239:246 arg1 neutral 261:267 neutral 261:267 neutral 261:267 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 1 64 dep branched 239:246 arg1 sialylated 249:258 sialylated 249:258 sialylated 249:258 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 1 65 theme capillary 170:178 arg1 electrophoresis 180:194 capillary electrophoresis 170:194 capillary electrophoresis 170:194 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 26544759 2 66 dep glycans 648:654 arg1 both 656:659 both 656:659 both 656:659 Background electrolytes without as well as with low and high molecular weight additives (ethylene glycol, linear polyacrylamide and poly(ethylene oxide)) were investigated for this phenomena in the temperature range of 20-50 °C. Glucose unit (GU) value shifts were observed with increasing temperature for the all IgG glycans both in additive-free and additive-containing background electrolytes, emphasizing the importance of tight temperature control during glycosylation analysis by capillary electrophoresis. 26544759 1 67 theme core 273:276 arg1 glycans 306:312 core fucosylated biantennary IgG glycans 273:312 core fucosylated biantennary IgG glycans 273:312 Temperature dependent differential migration shifts were studied in capillary electrophoresis between linear (maltooligosaccharides) and branched (sialylated, neutral and core fucosylated biantennary IgG glycans) carbohydrates. 27612524 3 0 theme glycosyl 505:512 arg1 analysis 522:529 glycosyl linkage analysis 505:529 glycosyl linkage analysis 505:529 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 1 theme syrup 593:597 arg1 polysaccharides 599:613 the maple syrup polysaccharides 583:613 the maple syrup polysaccharides 583:613 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 2 theme linkage 514:520 arg1 analysis 522:529 glycosyl linkage analysis 505:529 glycosyl linkage analysis 505:529 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 3 theme maple 587:591 arg1 polysaccharides 599:613 the maple syrup polysaccharides 583:613 the maple syrup polysaccharides 583:613 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 4 theme maple 344:348 arg1 syrup 350:354 maple syrup 344:354 maple syrup 344:354 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 0 5 from Detection 0:8 arg1 Syrup 58:62 Syrup 58:62 Syrup 58:62 Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup. 27612524 3 6 theme magnetic 544:551 arg1 resonance 553:561 nuclear magnetic resonance 536:561 nuclear magnetic resonance spectroscopy 536:574 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 7 theme broad 678:682 arg1 distribution 701:712 a broad molecular weight distribution 676:712 a broad molecular weight distribution 676:712 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 2 8 theme maple 301:305 arg1 syrup 307:311 maple syrup 301:311 maple syrup 301:311 During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. 27612524 3 9 theme resonance 553:561 arg1 spectroscopy 563:574 nuclear magnetic resonance spectroscopy 536:574 nuclear magnetic resonance spectroscopy 536:574 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 10 from sap 792:794 arg1 isolation 738:746 the first isolation 728:746 the first isolation of this prebiotic carbohydrate from a xylem sap 728:794 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 2 11 theme phytochemical 256:268 arg1 components 270:279 natural phytochemical components 248:279 natural phytochemical components 248:279 During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. 27612524 1 12 theme natural 112:118 arg1 syrup 71:75 Maple syrup 65:75 Maple syrup 65:75 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 12 theme natural 112:118 arg1 sweetener 120:128 a widely consumed plant-derived natural sweetener 80:128 a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species 80:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 4 13 theme carbohydrate 766:777 arg1 isolation 738:746 the first isolation 728:746 the first isolation of this prebiotic carbohydrate from a xylem sap 728:794 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 2 14 theme natural 248:254 arg1 components 270:279 natural phytochemical components 248:279 natural phytochemical components 248:279 During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. 27612524 5 15 theme rhamnogalacturonan 913:930 arg1 type 932:935 rhamnogalacturonan type I pectic polysaccharides 913:960 rhamnogalacturonan type I pectic polysaccharides 913:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 1 16 theme xylem 156:160 arg1 sap 162:164 xylem sap 156:164 xylem sap collected from certain maple (Acer) species 156:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 3 17 theme nuclear 536:542 arg1 resonance 553:561 nuclear magnetic resonance 536:561 nuclear magnetic resonance spectroscopy 536:574 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 1 18 theme Maple 65:69 arg1 syrup 71:75 Maple syrup 65:75 Maple syrup 65:75 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 18 theme Maple 65:69 arg1 sweetener 120:128 a widely consumed plant-derived natural sweetener 80:128 a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species 80:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 4 19 theme weight 694:699 arg1 distribution 701:712 a broad molecular weight distribution 676:712 a broad molecular weight distribution 676:712 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 20 theme ethanol 373:379 arg1 precipitation 381:393 ethanol precipitation 373:393 ethanol precipitation 373:393 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 21 theme prebiotic 756:764 arg1 carbohydrate 766:777 this prebiotic carbohydrate 751:777 this prebiotic carbohydrate 751:777 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 22 theme anion 410:414 arg1 chromatography 425:438 anion exchange chromatography 410:438 anion exchange chromatography 410:438 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 5 23 theme structural 842:851 arg1 similarity 853:862 structural similarity 842:862 structural similarity 842:862 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 5 24 with polysaccharides 821:835 arg1 similarity 853:862 structural similarity 842:862 structural similarity 842:862 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 1 25 theme certain 181:187 arg1 species 202:208 certain maple (Acer) species 181:208 certain maple (Acer) species 181:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 0 26 theme Inulin 13:18 arg1 Detection 0:8 Detection 0:8 Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup. 0:63 Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup. 27612524 1 27 theme maple 189:193 arg1 species 202:208 certain maple (Acer) species 181:208 certain maple (Acer) species 181:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 0 28 theme Prebiotic 23:31 arg1 Inulin 13:18 Inulin 13:18 Inulin 13:18 Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup. 27612524 0 28 theme Prebiotic 23:31 arg1 Polysaccharide 33:46 a Prebiotic Polysaccharide 21:46 a Prebiotic Polysaccharide 21:46 Detection of Inulin, a Prebiotic Polysaccharide, in Maple Syrup. 27612524 4 29 theme first 732:736 arg1 isolation 738:746 the first isolation 728:746 the first isolation of this prebiotic carbohydrate from a xylem sap 728:794 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 5 30 attach derived 900:906 arg2 arabinogalactans 883:898 arabinogalactans 883:898 arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides 883:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 5 30 attach derived 900:906 arg2 polysaccharides 821:835 two acidic polysaccharides 810:835 two acidic polysaccharides with structural similarity 810:862 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 5 30 attach derived 900:906 arg1 type 932:935 rhamnogalacturonan type I pectic polysaccharides 913:960 rhamnogalacturonan type I pectic polysaccharides 913:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 4 31 theme molecular 684:692 arg1 distribution 701:712 a broad molecular weight distribution 676:712 a broad molecular weight distribution 676:712 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 32 theme exchange 416:423 arg1 chromatography 425:438 anion exchange chromatography 410:438 anion exchange chromatography 410:438 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 4 33 theme xylem 786:790 arg1 sap 792:794 a xylem sap 784:794 a xylem sap 784:794 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 5 34 dep type 932:935 arg1 polysaccharides 946:960 pectic polysaccharides 939:960 rhamnogalacturonan type I pectic polysaccharides 913:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 1 35 theme Acer 196:199 arg1 species 202:208 certain maple (Acer) species 181:208 certain maple (Acer) species 181:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 36 link plant-derived 98:110 arg1 syrup 71:75 Maple syrup 65:75 Maple syrup 65:75 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 36 link plant-derived 98:110 arg1 sweetener 120:128 a widely consumed plant-derived natural sweetener 80:128 a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species 80:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 3 37 from syrup 350:354 arg1 components 328:337 The polymeric components 314:337 The polymeric components from maple syrup 314:354 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 5 38 theme acidic 814:819 arg1 arabinogalactans 883:898 arabinogalactans 883:898 arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides 883:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 5 38 theme acidic 814:819 arg1 polysaccharides 821:835 two acidic polysaccharides 810:835 two acidic polysaccharides with structural similarity 810:862 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 27612524 1 39 theme consumed 89:96 arg1 syrup 71:75 Maple syrup 65:75 Maple syrup 65:75 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 39 theme consumed 89:96 arg1 sweetener 120:128 a widely consumed plant-derived natural sweetener 80:128 a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species 80:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 4 40 with inulin 664:669 arg1 distribution 701:712 a broad molecular weight distribution 676:712 a broad molecular weight distribution 676:712 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 2 41 theme water 241:245 arg1 evaporation 226:236 thermal evaporation 218:236 thermal evaporation of water 218:245 During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. 27612524 1 42 theme plant-derived 98:110 arg1 syrup 71:75 Maple syrup 65:75 Maple syrup 65:75 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 1 42 theme plant-derived 98:110 arg1 sweetener 120:128 a widely consumed plant-derived natural sweetener 80:128 a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species 80:208 Maple syrup is a widely consumed plant-derived natural sweetener produced by concentrating xylem sap collected from certain maple (Acer) species. 27612524 4 43 theme neutral 620:626 arg1 inulin 664:669 inulin 664:669 inulin with a broad molecular weight distribution 664:712 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 4 43 theme neutral 620:626 arg1 polysaccharide 628:641 one neutral polysaccharide 616:641 one neutral polysaccharide 616:641 Among the maple syrup polysaccharides, one neutral polysaccharide was characterized as inulin with a broad molecular weight distribution, representing the first isolation of this prebiotic carbohydrate from a xylem sap. 27612524 3 44 theme composition 483:493 arg1 analysis 495:502 glycosyl composition analysis 474:502 glycosyl composition analysis 474:502 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 3 45 theme glycosyl 474:481 arg1 analysis 495:502 glycosyl composition analysis 474:502 glycosyl composition analysis 474:502 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 3 46 theme polymeric 318:326 arg1 components 328:337 The polymeric components 314:337 The polymeric components from maple syrup 314:354 The polymeric components from maple syrup were isolated by ethanol precipitation, dialysis, and anion exchange chromatography and structurally characterized by glycosyl composition analysis, glycosyl linkage analysis, and nuclear magnetic resonance spectroscopy. 27612524 2 47 theme thermal 218:224 arg1 evaporation 226:236 thermal evaporation 218:236 thermal evaporation of water 218:245 During thermal evaporation of water, natural phytochemical components are concentrated in maple syrup. 27612524 5 48 theme pectic 939:944 arg1 polysaccharides 946:960 pectic polysaccharides 939:960 rhamnogalacturonan type I pectic polysaccharides 913:960 In addition, two acidic polysaccharides with structural similarity were identified as arabinogalactans derived from rhamnogalacturonan type I pectic polysaccharides. 26086522 5 0 theme oligosaccharides 634:649 arg1 elucidation 619:629 Structural elucidation 608:629 Structural elucidation of oligosaccharides 608:649 Structural elucidation of oligosaccharides remains difficult and time-consuming. 26086522 1 1 theme proteins 167:174 arg1 modification 151:162 an important post-translational modification 119:162 an important post-translational modification of proteins present in the vast majority of human proteins 119:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 1 1 theme proteins 167:174 arg1 Glycosylation 102:114 Glycosylation 102:114 Glycosylation 102:114 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 4 2 theme abundant 565:572 arg1 N-glycans 574:582 many abundant N-glycans 560:582 many abundant N-glycans 560:582 Even now, the exact structures of many abundant N-glycans in serum are not known. 26086522 3 3 from genome 395:400 arg1 complexity 418:427 the general complexity 406:427 the general complexity of the structures 406:445 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 3 3 from genome 395:400 arg1 absence 350:356 the absence 346:356 the absence of a biosynthetic template as in the genome 346:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 7 4 theme glycoproteins 963:975 arg1 handful 946:952 a handful 944:952 a handful of serum glycoproteins 944:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 1 5 theme present 176:182 arg1 proteins 167:174 proteins 167:174 proteins present in the vast majority of human proteins 167:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 4 6 theme many 560:563 arg1 N-glycans 574:582 many abundant N-glycans 560:582 many abundant N-glycans 560:582 Even now, the exact structures of many abundant N-glycans in serum are not known. 26086522 10 7 used used 1281:1284 arg2 approach 1269:1276 This approach 1264:1276 This approach 1264:1276 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 1 8 from present 176:182 arg1 majority 196:203 the vast majority 187:203 the vast majority of human proteins 187:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 8 9 theme mass 1143:1146 arg1 spectrometry 1148:1159 accurate mass spectrometry 1134:1159 accurate mass spectrometry 1134:1159 The method employs primarily nanoflow liquid chromatography and accurate mass spectrometry. 26086522 0 10 theme Biological 79:88 arg1 Variations 90:99 Biological Variations 79:99 Biological Variations 79:99 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 10 11 theme several 1360:1366 arg1 individuals 1368:1378 several individuals 1360:1378 several individuals providing detailed variations in the abundances of the important N-glycans in blood 1360:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 2 12 theme biomarkers 277:286 arg1 sources 266:272 potentially new sources 250:272 potentially new sources of biomarkers and active targets of therapeutics and vaccines 250:334 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 2 12 theme biomarkers 277:286 arg1 they 241:244 they 241:244 they 241:244 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 4 13 theme N-glycans 574:582 arg1 structures 546:555 the exact structures 536:555 the exact structures of many abundant N-glycans in serum 536:591 Even now, the exact structures of many abundant N-glycans in serum are not known. 26086522 9 14 from profile 1194:1200 arg1 fluids 1226:1231 biological fluids 1215:1231 biological fluids with deep structural analysis 1215:1261 The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. 26086522 10 15 from blood 1458:1462 arg1 abundances 1417:1426 the abundances 1413:1426 the abundances of the important N-glycans in blood 1413:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 7 16 theme serum 957:961 arg1 glycoproteins 963:975 serum glycoproteins 957:975 serum glycoproteins 957:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 10 17 theme relative 1303:1310 arg1 abundances 1312:1321 the relative abundances 1299:1321 the relative abundances 1299:1321 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 1 18 attach present 176:182 arg2 proteins 167:174 proteins 167:174 proteins present in the vast majority of human proteins 167:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 1 18 attach present 176:182 arg1 majority 196:203 the vast majority 187:203 the vast majority of human proteins 187:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 5 19 theme Structural 608:617 arg1 elucidation 619:629 Structural elucidation 608:629 Structural elucidation of oligosaccharides 608:649 Structural elucidation of oligosaccharides remains difficult and time-consuming. 26086522 6 20 theme retention 800:808 arg1 times 810:814 retention times 800:814 retention times based on a glycan library 800:840 Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. 26086522 7 21 theme serum 848:852 arg1 library 861:867 This serum glycan library 843:867 This serum glycan library 843:867 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 3 22 theme template 376:383 arg1 complexity 418:427 the general complexity 406:427 the general complexity of the structures 406:445 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 3 22 theme template 376:383 arg1 absence 350:356 the absence 346:356 the absence of a biosynthetic template as in the genome 346:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 10 23 from N-glycans 1445:1453 arg1 blood 1458:1462 blood 1458:1462 blood 1458:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 2 24 theme therapeutics 310:321 arg1 biomarkers 277:286 biomarkers 277:286 biomarkers 277:286 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 2 24 theme therapeutics 310:321 arg1 targets 299:305 active targets 292:305 active targets 292:305 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 7 25 theme glycan 854:859 arg1 library 861:867 This serum glycan library 843:867 This serum glycan library 843:867 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 1 26 theme vast 191:194 arg1 majority 196:203 the vast majority 187:203 the vast majority of human proteins 187:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 4 27 from structures 546:555 arg1 serum 587:591 serum 587:591 serum 587:591 Even now, the exact structures of many abundant N-glycans in serum are not known. 26086522 4 28 theme exact 540:544 arg1 structures 546:555 the exact structures 536:555 the exact structures of many abundant N-glycans in serum 536:591 Even now, the exact structures of many abundant N-glycans in serum are not known. 26086522 10 29 from individuals 1368:1378 arg1 variations 1327:1336 variations 1327:1336 variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood 1327:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 10 29 from individuals 1368:1378 arg1 N-glycans 1345:1353 the N-glycans 1341:1353 the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood 1341:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 10 29 from individuals 1368:1378 arg1 abundances 1312:1321 the relative abundances 1299:1321 the relative abundances 1299:1321 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 10 30 theme N-glycans 1445:1453 arg1 abundances 1417:1426 the abundances 1413:1426 the abundances of the important N-glycans in blood 1413:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 3 31 theme general 410:416 arg1 complexity 418:427 the general complexity 406:427 the general complexity of the structures 406:445 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 0 32 theme In-Depth 13:20 arg1 Annotation 33:42 In-Depth Structural Annotation 13:42 In-Depth Structural Annotation of Human Serum Glycans 13:65 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 6 33 theme accurate 780:787 arg1 masses 789:794 their accurate masses 774:794 their accurate masses 774:794 Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. 26086522 2 34 theme targets 299:305 arg1 sources 266:272 potentially new sources 250:272 potentially new sources of biomarkers and active targets of therapeutics and vaccines 250:334 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 2 34 theme targets 299:305 arg1 they 241:244 they 241:244 they 241:244 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 10 35 theme important 1435:1443 arg1 N-glycans 1445:1453 the important N-glycans 1431:1453 the important N-glycans in blood 1431:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 3 36 theme biosynthetic 363:374 arg1 template 376:383 a biosynthetic template 361:383 a biosynthetic template as in the genome 361:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 7 37 theme complete 996:1003 arg1 structures 1017:1026 more than 170 complete and partial structures 982:1026 more than 170 complete and partial structures 982:1026 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 9 38 theme structural 1243:1252 arg1 analysis 1254:1261 deep structural analysis 1238:1261 deep structural analysis 1238:1261 The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. 26086522 2 39 theme active 292:297 arg1 targets 299:305 active targets 292:305 active targets 292:305 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 1 40 theme human 208:212 arg1 proteins 214:221 human proteins 208:221 human proteins 208:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 9 41 with fluids 1226:1231 arg1 analysis 1254:1261 deep structural analysis 1238:1261 deep structural analysis 1238:1261 The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. 26086522 7 42 theme whole 1057:1061 arg1 serum 1063:1067 whole serum 1057:1067 whole serum 1057:1067 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 1 43 theme proteins 214:221 arg1 majority 196:203 the vast majority 187:203 the vast majority of human proteins 187:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 0 44 theme Structural 22:31 arg1 Annotation 33:42 In-Depth Structural Annotation 13:42 In-Depth Structural Annotation of Human Serum Glycans 13:65 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 7 45 gly glycoproteins 963:975 arg1 glycoproteins 963:975 serum glycoproteins 957:975 serum glycoproteins 957:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 3 46 from absence 350:356 arg1 genome 395:400 the genome 391:400 the genome 391:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 7 47 theme partial 1009:1015 arg1 structures 1017:1026 more than 170 complete and partial structures 982:1026 more than 170 complete and partial structures 982:1026 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 3 48 theme structures 436:445 arg1 complexity 418:427 the general complexity 406:427 the general complexity of the structures 406:445 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 3 48 theme structures 436:445 arg1 absence 350:356 the absence 346:356 the absence of a biosynthetic template as in the genome 346:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 0 49 theme Human 47:51 arg1 Glycans 59:65 Human Serum Glycans 47:65 Human Serum Glycans 47:65 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 10 50 from variations 1327:1336 arg1 N-glycans 1345:1353 the N-glycans 1341:1353 the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood 1341:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 10 50 from variations 1327:1336 arg1 individuals 1368:1378 several individuals 1360:1378 several individuals providing detailed variations in the abundances of the important N-glycans in blood 1360:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 7 51 contain has 978:980 arg2 structures 1017:1026 more than 170 complete and partial structures 982:1026 more than 170 complete and partial structures 982:1026 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 7 51 contain has 978:980 arg1 library 861:867 This serum glycan library 843:867 This serum glycan library 843:867 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 10 52 from abundances 1312:1321 arg1 N-glycans 1345:1353 the N-glycans 1341:1353 the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood 1341:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 10 52 from abundances 1312:1321 arg1 individuals 1368:1378 several individuals 1360:1378 several individuals providing detailed variations in the abundances of the important N-glycans in blood 1360:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 3 53 from template 376:383 arg1 genome 395:400 the genome 391:400 the genome 391:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 10 54 from abundances 1417:1426 arg1 blood 1458:1462 blood 1458:1462 blood 1458:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 8 55 theme liquid 1108:1113 arg1 chromatography 1115:1128 nanoflow liquid chromatography 1099:1128 nanoflow liquid chromatography 1099:1128 The method employs primarily nanoflow liquid chromatography and accurate mass spectrometry. 26086522 3 56 theme comprehensive 491:503 arg1 analysis 516:523 comprehensive structural analysis 491:523 comprehensive structural analysis 491:523 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 3 57 theme methods 479:485 arg1 development 464:474 the development 460:474 the development of methods for comprehensive structural analysis 460:523 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 10 58 from variations 1399:1408 arg1 abundances 1417:1426 the abundances 1413:1426 the abundances of the important N-glycans in blood 1413:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 9 59 theme deep 1238:1241 arg1 analysis 1254:1261 deep structural analysis 1238:1261 deep structural analysis 1238:1261 The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. 26086522 8 60 theme nanoflow 1099:1106 arg1 chromatography 1115:1128 nanoflow liquid chromatography 1099:1128 nanoflow liquid chromatography 1099:1128 The method employs primarily nanoflow liquid chromatography and accurate mass spectrometry. 26086522 9 61 theme biological 1215:1224 arg1 fluids 1226:1231 biological fluids 1215:1231 biological fluids with deep structural analysis 1215:1261 The method allows us to readily profile N-glycans in biological fluids with deep structural analysis. 26086522 2 62 theme new 262:264 arg1 sources 266:272 potentially new sources 250:272 potentially new sources of biomarkers and active targets of therapeutics and vaccines 250:334 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 2 62 theme new 262:264 arg1 they 241:244 they 241:244 they 241:244 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 6 63 theme N-glycan 748:755 arg1 structures 757:766 released N-glycan structures 739:766 released N-glycan structures using their accurate masses 739:794 Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. 26086522 1 64 theme important 122:130 arg1 modification 151:162 an important post-translational modification 119:162 an important post-translational modification of proteins present in the vast majority of human proteins 119:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 1 64 theme important 122:130 arg1 Glycosylation 102:114 Glycosylation 102:114 Glycosylation 102:114 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 10 65 theme detailed 1390:1397 arg1 variations 1399:1408 detailed variations 1390:1408 detailed variations in the abundances of the important N-glycans in blood 1390:1462 This approach is used to determine the relative abundances and variations in the N-glycans from several individuals providing detailed variations in the abundances of the important N-glycans in blood. 26086522 6 66 theme released 739:746 arg1 structures 757:766 released N-glycan structures 739:766 released N-glycan structures using their accurate masses 739:794 Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. 26086522 1 67 theme post-translational 132:149 arg1 modification 151:162 an important post-translational modification 119:162 an important post-translational modification of proteins present in the vast majority of human proteins 119:221 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 1 67 theme post-translational 132:149 arg1 Glycosylation 102:114 Glycosylation 102:114 Glycosylation 102:114 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 7 68 theme previous 900:907 arg1 one 909:911 a previous one 898:911 a previous one covering glycans released from a handful of serum glycoproteins 898:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 2 69 theme vaccines 327:334 arg1 biomarkers 277:286 biomarkers 277:286 biomarkers 277:286 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 2 69 theme vaccines 327:334 arg1 targets 299:305 active targets 292:305 active targets 292:305 For this reason, they are potentially new sources of biomarkers and active targets of therapeutics and vaccines. 26086522 0 70 theme Glycans 59:65 arg1 Annotation 33:42 In-Depth Structural Annotation 13:42 In-Depth Structural Annotation of Human Serum Glycans 13:65 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 6 71 theme glycan 827:832 arg1 library 834:840 a glycan library 825:840 a glycan library 825:840 Here, we introduce a means of rapidly identifying released N-glycan structures using their accurate masses and retention times based on a glycan library. 26086522 8 72 theme accurate 1134:1141 arg1 spectrometry 1148:1159 accurate mass spectrometry 1134:1159 accurate mass spectrometry 1134:1159 The method employs primarily nanoflow liquid chromatography and accurate mass spectrometry. 26086522 1 73 from majority 196:203 arg1 present 176:182 present 176:182 present 176:182 Glycosylation is an important post-translational modification of proteins present in the vast majority of human proteins. 26086522 3 74 theme structural 505:514 arg1 analysis 516:523 comprehensive structural analysis 491:523 comprehensive structural analysis 491:523 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 26086522 0 75 theme Serum 53:57 arg1 Glycans 59:65 Human Serum Glycans 47:65 Human Serum Glycans 47:65 A Method for In-Depth Structural Annotation of Human Serum Glycans That Yields Biological Variations. 26086522 7 76 attach released 930:937 arg2 glycans 922:928 glycans 922:928 glycans released from a handful of serum glycoproteins 922:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 7 76 attach released 930:937 arg1 handful 946:952 a handful 944:952 a handful of serum glycoproteins 944:975 This serum glycan library, significantly expanded from a previous one covering glycans released from a handful of serum glycoproteins, has more than 170 complete and partial structures and constructed instead from whole serum. 26086522 3 77 from complexity 418:427 arg1 genome 395:400 the genome 391:400 the genome 391:400 However, the absence of a biosynthetic template as in the genome and the general complexity of the structures have limited the development of methods for comprehensive structural analysis. 24299865 6 0 from C-2 852:854 arg1 DHPD1 888:892 DHPD1 888:892 DHPD1 888:892 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 4 1 theme substitution 536:547 arg1 degree 526:531 a high degree 519:531 a high degree of substitution (DS) 519:552 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 24299865 5 2 theme reaction 682:689 arg1 fixed 705:709 fixed 705:709 fixed 705:709 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 2 theme reaction 682:689 arg1 condition 691:699 the reaction condition 678:699 the reaction condition 678:699 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 6 3 theme NMR 799:801 arg1 spectra 803:809 NMR spectra 799:809 NMR spectra 799:809 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 2 4 theme Da 291:292 arg1 weight 270:275 molecular weight 260:275 molecular weight of 3.2 × 10(3)Da 260:292 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 5 5 from ratio 774:778 arg1 fixed 705:709 fixed 705:709 fixed 705:709 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 5 from ratio 774:778 arg1 condition 691:699 the reaction condition 678:699 the reaction condition 678:699 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 1 6 with plant 162:166 arg1 values 190:195 high nutritional values 173:195 high nutritional values 173:195 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 1 6 with plant 162:166 arg1 functions 208:216 health functions 201:216 health functions 201:216 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 5 7 theme Pyr 783:785 arg1 temperature 723:733 reaction temperature 60°C 714:738 reaction temperature 60°C 714:738 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 7 theme Pyr 783:785 arg1 min 759:761 reaction time 160 min 741:761 reaction time 160 min 741:761 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 7 theme Pyr 783:785 arg1 ratio 774:778 volume ratio 767:778 volume ratio of Pyr to CSA 2:1 767:796 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 1 8 theme health 201:206 arg1 functions 208:216 health functions 201:216 health functions 201:216 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 0 9 from abilities 48:56 arg1 huoshanense 93:103 Dendrobium huoshanense 82:103 Dendrobium huoshanense 82:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 4 10 theme surface 615:621 arg1 methodology 623:633 response surface methodology 606:633 response surface methodology 606:633 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 24299865 7 11 theme % 992:992 arg1 ability 977:983 the inhibitory ability 962:983 the inhibitory ability 962:983 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 1 12 theme Dendrobium 106:115 arg1 plant 162:166 an important edible-medicinal plant 132:166 an important edible-medicinal plant with high nutritional values and health functions 132:216 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 1 12 theme Dendrobium 106:115 arg1 huoshanense 117:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 2 13 dep 10 286:287 arg1 3 289:289 3 289:289 3 289:289 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 0 14 theme Sulfated 0:7 arg1 modification 9:20 Sulfated modification 0:20 Sulfated modification 0:20 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 4 15 theme high 521:524 arg1 degree 526:531 a high degree 519:531 a high degree of substitution (DS) 519:552 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 24299865 3 16 theme Chlorosulfonic 409:422 arg1 acid-pyridine 424:436 Chlorosulfonic acid-pyridine 409:436 Chlorosulfonic acid-pyridine (CSA-Pyr) method 409:453 Chlorosulfonic acid-pyridine (CSA-Pyr) method was performed to modify the structure of DHPD1. 24299865 3 16 theme Chlorosulfonic 409:422 arg1 CSA-Pyr 439:445 CSA-Pyr 439:445 CSA-Pyr 439:445 Chlorosulfonic acid-pyridine (CSA-Pyr) method was performed to modify the structure of DHPD1. 24299865 5 17 theme volume 767:772 arg1 ratio 774:778 volume ratio 767:778 volume ratio of Pyr to CSA 2:1 767:796 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 18 theme time 750:753 arg1 min 759:761 reaction time 160 min 741:761 reaction time 160 min 741:761 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 3 19 theme DHPD1 496:500 arg1 structure 483:491 the structure 479:491 the structure of DHPD1 479:500 Chlorosulfonic acid-pyridine (CSA-Pyr) method was performed to modify the structure of DHPD1. 24299865 7 20 theme same 1075:1078 arg1 dosage 1080:1085 the same dosage 1071:1085 the same dosage 1071:1085 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 7 21 theme inhibitory 966:975 arg1 ability 977:983 the inhibitory ability 962:983 the inhibitory ability 962:983 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 0 22 from huoshanense 93:103 arg1 abilities 48:56 antiglycation abilities 34:56 antiglycation abilities of polysaccharides from Dendrobium huoshanense 34:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 0 22 from huoshanense 93:103 arg1 polysaccharides 61:75 polysaccharides 61:75 polysaccharides from Dendrobium huoshanense 61:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 4 23 theme response 606:613 arg1 methodology 623:633 response surface methodology 606:633 response surface methodology 606:633 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 24299865 0 24 theme antiglycation 34:46 arg1 abilities 48:56 antiglycation abilities 34:56 antiglycation abilities of polysaccharides from Dendrobium huoshanense 34:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 6 25 theme residues 876:883 arg1 C-6 860:862 C-6 860:862 C-6 860:862 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 6 25 theme residues 876:883 arg1 C-2 852:854 C-2 852:854 C-2 852:854 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 5 26 theme reaction 741:748 arg1 min 759:761 reaction time 160 min 741:761 reaction time 160 min 741:761 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 2 27 theme molecular 260:268 arg1 weight 270:275 molecular weight 260:275 molecular weight of 3.2 × 10(3)Da 260:292 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 7 28 theme sulfated 928:935 arg1 DHPD1 937:941 the sulfated DHPD1 924:941 the sulfated DHPD1 at 1.0mg/mL 924:953 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 5 29 theme CSA 790:792 arg1 2:1 794:796 CSA 2:1 790:796 CSA 2:1 790:796 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 0 30 theme polysaccharides 61:75 arg1 abilities 48:56 antiglycation abilities 34:56 antiglycation abilities of polysaccharides from Dendrobium huoshanense 34:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 6 31 theme glycosyl 867:874 arg1 residues 876:883 glycosyl residues 867:883 glycosyl residues 867:883 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 5 32 theme maximum 640:646 arg1 DS 648:649 The maximum DS 636:649 The maximum DS of 1.473 636:658 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 3 33 theme acid-pyridine 424:436 arg1 method 448:453 Chlorosulfonic acid-pyridine (CSA-Pyr) method 409:453 Chlorosulfonic acid-pyridine (CSA-Pyr) method 409:453 Chlorosulfonic acid-pyridine (CSA-Pyr) method was performed to modify the structure of DHPD1. 24299865 5 34 from min 759:761 arg1 fixed 705:709 fixed 705:709 fixed 705:709 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 34 from min 759:761 arg1 condition 691:699 the reaction condition 678:699 the reaction condition 678:699 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 1 35 theme important 135:143 arg1 plant 162:166 an important edible-medicinal plant 132:166 an important edible-medicinal plant with high nutritional values and health functions 132:216 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 1 35 theme important 135:143 arg1 huoshanense 117:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 6 36 from C-6 860:862 arg1 DHPD1 888:892 DHPD1 888:892 DHPD1 888:892 NMR spectra revealed that this sulfation occurred to C-2 and C-6 of glycosyl residues in DHPD1. 24299865 5 37 from temperature 723:733 arg1 fixed 705:709 fixed 705:709 fixed 705:709 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 5 37 from temperature 723:733 arg1 condition 691:699 the reaction condition 678:699 the reaction condition 678:699 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 1 38 theme edible-medicinal 145:160 arg1 plant 162:166 an important edible-medicinal plant 132:166 an important edible-medicinal plant with high nutritional values and health functions 132:216 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 1 38 theme edible-medicinal 145:160 arg1 huoshanense 117:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense 106:127 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 1 39 theme high 173:176 arg1 values 190:195 high nutritional values 173:195 high nutritional values 173:195 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 2 40 dep D. 313:314 arg1 composed 346:353 composed 346:353 was mainly composed of glucose, arabinose, galactose, mannose and xylose 335:406 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 2 40 dep D. 313:314 arg1 huoshanense 316:326 D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose 313:406 huoshanense 316:326 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 4 41 theme sulfated 555:562 arg1 conditions 577:586 sulfated modification conditions 555:586 sulfated modification conditions 555:586 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 24299865 5 42 theme 1.473 654:658 arg1 DS 648:649 The maximum DS 636:649 The maximum DS of 1.473 636:658 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 7 43 from 1.0mg/mL 946:953 arg1 DHPD1 937:941 the sulfated DHPD1 924:941 the sulfated DHPD1 at 1.0mg/mL 924:953 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 0 44 theme Dendrobium 82:91 arg1 huoshanense 93:103 Dendrobium huoshanense 82:103 Dendrobium huoshanense 82:103 Sulfated modification can enhance antiglycation abilities of polysaccharides from Dendrobium huoshanense. 24299865 2 45 theme homogenous 221:230 arg1 polysaccharide 232:245 A homogenous polysaccharide 219:245 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da 219:292 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 2 45 theme homogenous 221:230 arg1 DHPD1 248:252 DHPD1 248:252 DHPD1 248:252 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 7 46 theme incubation 912:921 arg1 days 904:907 28 days 901:907 28 days of incubation 901:921 After 28 days of incubation, the sulfated DHPD1 at 1.0mg/mL showed the inhibitory ability of 58.5%, which increased by 16.2% and 52.5% than that of aminoguanidine and DHPD1 at the same dosage. 24299865 2 47 with polysaccharide 232:245 arg1 weight 270:275 molecular weight 260:275 molecular weight of 3.2 × 10(3)Da 260:292 A homogenous polysaccharide (DHPD1) with molecular weight of 3.2 × 10(3)Da was extracted from D. huoshanense, which was mainly composed of glucose, arabinose, galactose, mannose and xylose. 24299865 5 48 theme reaction 714:721 arg1 temperature 723:733 reaction temperature 60°C 714:738 reaction temperature 60°C 714:738 The maximum DS of 1.473 was obtained when the reaction condition was fixed at reaction temperature 60°C, reaction time 160 min and volume ratio of Pyr to CSA 2:1. 24299865 1 49 theme nutritional 178:188 arg1 values 190:195 high nutritional values 173:195 high nutritional values 173:195 Dendrobium huoshanense is an important edible-medicinal plant with high nutritional values and health functions. 24299865 4 50 theme modification 564:575 arg1 conditions 577:586 sulfated modification conditions 555:586 sulfated modification conditions 555:586 In order to get a high degree of substitution (DS), sulfated modification conditions were optimized by response surface methodology. 25974358 1 0 theme disease-related 151:165 arg1 mechanisms 167:176 disease-related mechanisms 151:176 disease-related mechanisms 151:176 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 5 1 theme therapeutics 990:1001 arg1 development 928:938 the development 924:938 the development of new and more effective, synthetic glycan-based therapeutics and vaccines 924:1014 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 3 2 theme carbohydrate 542:553 arg1 structures 555:564 the complex carbohydrate structures 530:564 the complex carbohydrate structures 530:564 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 4 3 from interest 770:777 arg1 carbohydrates 819:831 carbohydrates 819:831 carbohydrates 819:831 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 5 4 theme glycan-based 977:988 arg1 therapeutics 990:1001 synthetic glycan-based therapeutics 967:1001 synthetic glycan-based therapeutics 967:1001 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 0 5 from Glycans 0:6 arg1 Chemistry 21:29 Medicinal Chemistry 11:29 Medicinal Chemistry 11:29 Glycans in Medicinal Chemistry: An Underexploited Resource. 25974358 3 6 theme functions 587:595 arg1 understanding 513:525 A more detailed understanding 497:525 A more detailed understanding of the complex carbohydrate structures and their associated functions 497:595 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 3 7 theme complex 534:540 arg1 structures 555:564 the complex carbohydrate structures 530:564 the complex carbohydrate structures 530:564 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 4 8 theme community 806:814 arg1 chemistry 796:804 the medicinal chemistry community 782:814 the medicinal chemistry community 782:814 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 5 9 theme vaccines 1007:1014 arg1 development 928:938 the development 924:938 the development of new and more effective, synthetic glycan-based therapeutics and vaccines 924:1014 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 1 10 theme biological 64:73 arg1 relevance 75:83 The biological relevance 60:83 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, 60:177 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 2 11 theme biological 275:284 arg1 roles 286:290 their important biological roles 259:290 their important biological roles 259:290 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 4 12 theme chemical 732:739 arg1 glycobiology 741:752 chemical glycobiology 732:752 chemical glycobiology 732:752 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 2 13 theme non-drug-like 471:483 arg1 properties 485:494 their non-drug-like properties 465:494 their non-drug-like properties 465:494 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 5 14 theme effective 956:964 arg1 therapeutics 990:1001 synthetic glycan-based therapeutics 967:1001 synthetic glycan-based therapeutics 967:1001 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 1 15 theme glycans 88:94 arg1 relevance 75:83 The biological relevance 60:83 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, 60:177 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 2 16 theme recognition 317:327 arg1 elements 329:336 molecular recognition elements 307:336 molecular recognition elements 307:336 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 1 17 theme attractive 190:199 arg1 targets 201:207 attractive targets 190:207 attractive targets for a wide range of medical applications 190:248 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 5 18 theme discovery 902:910 arg1 efforts 912:918 drug discovery efforts 897:918 drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines 897:1014 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 2 19 theme molecular 307:315 arg1 elements 329:336 molecular recognition elements 307:336 molecular recognition elements 307:336 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 3 20 theme glycan-based 641:652 arg1 pharmaceuticals 654:668 new glycan-based pharmaceuticals 637:668 new glycan-based pharmaceuticals 637:668 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 2 21 theme structure-activity 429:446 arg1 correlations 448:459 structure-activity correlations 429:459 structure-activity correlations 429:459 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 0 22 theme Medicinal 11:19 arg1 Chemistry 21:29 Medicinal Chemistry 11:29 Medicinal Chemistry 11:29 Glycans in Medicinal Chemistry: An Underexploited Resource. 25974358 5 23 theme synthetic 967:975 arg1 therapeutics 990:1001 synthetic glycan-based therapeutics 967:1001 synthetic glycan-based therapeutics 967:1001 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 4 24 theme medicinal 786:794 arg1 chemistry 796:804 the medicinal chemistry community 782:814 the medicinal chemistry community 782:814 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 0 25 dep Resource 50:57 arg1 Glycans 0:6 Glycans 0:6 Glycans in Medicinal Chemistry 0:29 Glycans in Medicinal Chemistry: An Underexploited Resource. 25974358 3 26 theme pharmaceuticals 654:668 arg1 development 622:632 the development 618:632 the development of new glycan-based pharmaceuticals 618:668 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 1 27 theme wide 215:218 arg1 range 220:224 a wide range 213:224 a wide range of medical applications 213:248 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 4 28 theme significant 678:688 arg1 progress 690:697 Recent significant progress 671:697 Recent significant progress in oligosaccharide synthesis and chemical glycobiology 671:752 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 4 29 theme chemistry 796:804 arg1 interest 770:777 the interest 766:777 the interest of the medicinal chemistry community in carbohydrates 766:831 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 0 30 theme Underexploited 35:48 arg1 Resource 50:57 An Underexploited Resource 32:57 An Underexploited Resource 32:57 Glycans in Medicinal Chemistry: An Underexploited Resource. 25974358 3 31 theme new 637:639 arg1 pharmaceuticals 654:668 new glycan-based pharmaceuticals 637:668 new glycan-based pharmaceuticals 637:668 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 1 32 theme key 112:114 arg1 processes 130:138 key physiological processes 112:138 key physiological processes 112:138 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 1 32 theme key 112:114 arg1 mechanisms 167:176 disease-related mechanisms 151:176 disease-related mechanisms 151:176 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 2 33 theme important 265:273 arg1 roles 286:290 their important biological roles 259:290 their important biological roles 259:290 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 1 34 theme physiological 116:128 arg1 processes 130:138 key physiological processes 112:138 key physiological processes 112:138 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 1 34 theme physiological 116:128 arg1 mechanisms 167:176 disease-related mechanisms 151:176 disease-related mechanisms 151:176 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 4 35 theme Recent 671:676 arg1 progress 690:697 Recent significant progress 671:697 Recent significant progress in oligosaccharide synthesis and chemical glycobiology 671:752 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 3 36 theme associated 576:585 arg1 functions 587:595 their associated functions 570:595 their associated functions 570:595 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 1 37 theme processes 130:138 arg1 mediators 99:107 mediators 99:107 mediators of key physiological processes, including disease-related mechanisms, 99:177 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 3 38 theme detailed 504:511 arg1 understanding 513:525 A more detailed understanding 497:525 A more detailed understanding of the complex carbohydrate structures and their associated functions 497:595 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 4 39 from progress 690:697 arg1 synthesis 718:726 oligosaccharide synthesis 702:726 oligosaccharide synthesis 702:726 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 4 39 from progress 690:697 arg1 glycobiology 741:752 chemical glycobiology 732:752 chemical glycobiology 732:752 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 5 40 theme new 943:945 arg1 therapeutics 990:1001 synthetic glycan-based therapeutics 967:1001 synthetic glycan-based therapeutics 967:1001 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 4 41 theme oligosaccharide 702:716 arg1 synthesis 718:726 oligosaccharide synthesis 702:726 oligosaccharide synthesis 702:726 Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. 25974358 3 42 theme structures 555:564 arg1 understanding 513:525 A more detailed understanding 497:525 A more detailed understanding of the complex carbohydrate structures and their associated functions 497:595 A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan-based pharmaceuticals. 25974358 1 43 theme medical 229:235 arg1 applications 237:248 medical applications 229:248 medical applications 229:248 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 25974358 2 44 theme correlations 448:459 arg1 scarcity 417:424 the scarcity 413:424 the scarcity of structure-activity correlations 413:459 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 2 44 theme correlations 448:459 arg1 properties 485:494 their non-drug-like properties 465:494 their non-drug-like properties 465:494 Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure-activity correlations and their non-drug-like properties. 25974358 5 45 theme drug 897:900 arg1 efforts 912:918 drug discovery efforts 897:918 drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines 897:1014 This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan-based therapeutics and vaccines. 25974358 1 46 theme applications 237:248 arg1 range 220:224 a wide range 213:224 a wide range of medical applications 213:248 The biological relevance of glycans as mediators of key physiological processes, including disease-related mechanisms, makes them attractive targets for a wide range of medical applications. 28837324 1 0 theme critical 293:300 arg1 threat 311:316 the critical hospital threat 289:316 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 4 1 theme donors 860:865 arg1 glycosylation 826:838 the glycosylation 822:838 the glycosylation of the Pse glycosyl donors 822:865 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 2 2 theme 11 530:531 arg1 % 532:532 % 532:532 % 532:532 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 4 3 dep aeruginosa 1009:1018 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 4 4 theme 2→4 1063:1065 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 5 theme cell 211:214 arg1 glycans 224:230 important cell surface glycans 201:230 important cell surface glycans 201:230 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 6 theme functionalized 434:447 arg1 derivatives 449:459 its functionalized derivatives 430:459 its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 430:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 1 7 theme hospital 302:309 arg1 threat 311:316 the critical hospital threat 289:316 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 3 8 theme glycine 616:622 arg1 reaction 662:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction 593:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton 593:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 1 9 theme nonulosonic 131:141 arg1 acid 143:146 a nonulosonic acid 129:146 a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa 129:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 9 theme nonulosonic 131:141 arg1 acid 115:118 Pseudaminic acid 103:118 Pseudaminic acid (Pse) 103:124 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 10 theme surface 216:222 arg1 glycans 224:230 important cell surface glycans 201:230 important cell surface glycans 201:230 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 0 11 theme Pseudaminic 85:95 arg1 Acid 97:100 Pseudaminic Acid 85:100 Pseudaminic Acid 85:100 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 4 12 theme -β-Xyl- 1067:1073 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 13 theme glycans 224:230 arg1 component 188:196 a component 186:196 a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa 186:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 14 theme methyl 504:509 arg1 ester 511:515 easily available Cbz-l-allo-threonine methyl ester 466:515 easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 466:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 4 15 theme glycosyl 851:858 arg1 donors 860:865 the Pse glycosyl donors 843:865 the Pse glycosyl donors 843:865 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 4 16 theme aeruginosa 1009:1018 arg1 synthesis 993:1001 the total synthesis 983:1001 the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 983:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 2 17 theme Cbz-l-allo-threonine 483:502 arg1 ester 511:515 easily available Cbz-l-allo-threonine methyl ester 466:515 easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 466:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 2 18 dep de 401:402 arg1 novo 404:407 novo 404:407 novo 404:407 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 0 19 theme Acid 97:100 arg1 Synthesis 72:80 de Novo Synthesis 64:80 de Novo Synthesis of Pseudaminic Acid 64:100 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 3 20 theme 1,3-anti-diamino 685:700 arg1 skeleton 702:709 the 1,3-anti-diamino skeleton 681:709 the 1,3-anti-diamino skeleton 681:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 4 21 theme pilin 1025:1029 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 22 theme unique 148:153 arg1 acid 143:146 a nonulosonic acid 129:146 a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa 129:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 22 theme unique 148:153 arg1 acid 115:118 Pseudaminic acid 103:118 Pseudaminic acid (Pse) 103:124 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 23 theme synthesis 409:417 arg1 development 364:374 the development 360:374 the development of a facile and scalable de novo synthesis of Pse 360:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 2 23 theme synthesis 409:417 arg1 derivatives 449:459 its functionalized derivatives 430:459 its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 430:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 4 24 theme total 987:991 arg1 synthesis 993:1001 the total synthesis 983:1001 the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 983:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 4 25 theme Pse 847:849 arg1 donors 860:865 the Pse glycosyl donors 843:865 the Pse glycosyl donors 843:865 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 3 26 theme Fukuyama 728:735 arg1 reduction 737:745 the Fukuyama reduction 724:745 the Fukuyama reduction 724:745 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 0 27 theme Total 0:4 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 0:101 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 1 28 theme glycoproteins 236:248 arg1 component 188:196 a component 186:196 a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa 186:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 29 theme bacterial 158:166 arg1 species 168:174 bacterial species 158:174 bacterial species 158:174 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 30 theme de 401:402 arg1 synthesis 409:417 a facile and scalable de novo synthesis 379:417 a facile and scalable de novo synthesis of Pse 379:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 1 31 dep threat 311:316 arg1 aeruginosa 330:339 Pseudomonas aeruginosa 318:339 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 4 32 theme glycosylation 918:930 arg1 diastereoselectivity 932:951 its glycosylation diastereoselectivity 914:951 its glycosylation diastereoselectivity 914:951 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 0 33 dep de 64:65 arg1 Novo 67:70 Novo 67:70 Novo 67:70 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 4 34 gly glycosylation 826:838 arg1 donors 860:865 the Pse glycosyl donors 843:865 the Pse glycosyl donors 843:865 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 2 35 theme scalable 392:399 arg1 synthesis 409:417 a facile and scalable de novo synthesis 379:417 a facile and scalable de novo synthesis of Pse 379:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 1 36 theme various 253:259 arg1 threat 311:316 the critical hospital threat 289:316 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 36 theme various 253:259 arg1 species 272:278 various pathogenic species 253:278 various pathogenic species 253:278 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 37 theme facile 381:386 arg1 synthesis 409:417 a facile and scalable de novo synthesis 379:417 a facile and scalable de novo synthesis of Pse 379:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 1 38 theme pathogenic 261:270 arg1 threat 311:316 the critical hospital threat 289:316 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 38 theme pathogenic 261:270 arg1 species 272:278 various pathogenic species 253:278 various pathogenic species 253:278 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 39 dep ester 511:515 arg1 steps 521:525 16 steps 518:525 16 steps in 11% yield 518:538 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 3 40 theme diastereoselective 597:614 arg1 reaction 662:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction 593:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton 593:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 3 41 theme key 546:548 arg1 reactions 550:558 The key reactions 542:558 The key reactions in our de novo synthesis 542:583 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 3 42 from reactions 550:558 arg1 synthesis 575:583 our de novo synthesis 563:583 our de novo synthesis 563:583 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 3 43 dep de 567:568 arg1 novo 570:573 novo 570:573 novo 570:573 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 0 44 theme aeruginosa 31:40 arg1 Glycan 53:58 Pseudomonas aeruginosa 1244 Pilin Glycan 19:58 Pseudomonas aeruginosa 1244 Pilin Glycan 19:58 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 3 45 theme indium-mediated 755:769 arg1 allylation 784:793 the indium-mediated Barbier-type allylation 751:793 the indium-mediated Barbier-type allylation 751:793 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 2 46 theme available 473:481 arg1 ester 511:515 easily available Cbz-l-allo-threonine methyl ester 466:515 easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 466:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 0 47 theme Glycan 53:58 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 0:101 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 3 48 theme Barbier-type 771:782 arg1 allylation 784:793 the indium-mediated Barbier-type allylation 751:793 the indium-mediated Barbier-type allylation 751:793 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 4 49 theme structural 886:895 arg1 determinants 897:908 the structural determinants 882:908 the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 882:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 50 gly glycoproteins 236:248 arg1 glycoproteins 236:248 glycoproteins 236:248 glycoproteins 236:248 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 51 theme Pseudaminic 103:113 arg1 acid 143:146 a nonulosonic acid 129:146 a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa 129:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 51 theme Pseudaminic 103:113 arg1 Pse 121:123 Pse 121:123 Pse 121:123 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 51 theme Pseudaminic 103:113 arg1 acid 115:118 Pseudaminic acid 103:118 Pseudaminic acid (Pse) 103:124 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 0 52 theme Pilin 47:51 arg1 Glycan 53:58 Pseudomonas aeruginosa 1244 Pilin Glycan 19:58 Pseudomonas aeruginosa 1244 Pilin Glycan 19:58 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 3 53 theme isonitrile-based 634:649 arg1 reaction 662:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction 593:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton 593:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 2 54 theme % 532:532 arg1 yield 534:538 11% yield 530:538 11% yield 530:538 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 3 55 theme aldol-type 651:660 arg1 reaction 662:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction 593:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton 593:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 3 56 theme de 567:568 arg1 synthesis 575:583 our de novo synthesis 563:583 our de novo synthesis 563:583 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 4 57 theme 1→3 1075:1077 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 58 from component 188:196 arg1 threat 311:316 the critical hospital threat 289:316 the critical hospital threat Pseudomonas aeruginosa 289:339 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 1 58 from component 188:196 arg1 species 272:278 various pathogenic species 253:278 various pathogenic species 253:278 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28837324 2 59 from ester 511:515 arg1 development 364:374 the development 360:374 the development of a facile and scalable de novo synthesis of Pse 360:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 2 59 from ester 511:515 arg1 derivatives 449:459 its functionalized derivatives 430:459 its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield) 430:539 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 4 60 theme α-5NβOHC47NFmPse- 1045:1061 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 0 61 theme de 64:65 arg1 Synthesis 72:80 de Novo Synthesis 64:80 de Novo Synthesis of Pseudaminic Acid 64:100 Total Synthesis of Pseudomonas aeruginosa 1244 Pilin Glycan via de Novo Synthesis of Pseudaminic Acid. 28837324 3 62 theme thioester 624:632 arg1 reaction 662:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction 593:669 the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton 593:709 The key reactions in our de novo synthesis involve the diastereoselective glycine thioester isonitrile-based aldol-type reaction to create the 1,3-anti-diamino skeleton, followed by the Fukuyama reduction and the indium-mediated Barbier-type allylation. 28837324 2 63 theme Pse 422:424 arg1 synthesis 409:417 a facile and scalable de novo synthesis 379:417 a facile and scalable de novo synthesis of Pse 379:424 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 2 64 from steps 521:525 arg1 yield 534:538 11% yield 530:538 11% yield 530:538 Herein we present the development of a facile and scalable de novo synthesis of Pse and its functionalized derivatives from easily available Cbz-l-allo-threonine methyl ester (16 steps in 11% yield). 28837324 4 65 theme trisaccharide 1031:1043 arg1 -FucNAc 1079:1085 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1020:1085 P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc 1006:1085 Moreover, we have studied the glycosylation of the Pse glycosyl donors and identified the structural determinants for its glycosylation diastereoselectivity, which enabled us to complete the total synthesis of P. aeruginosa 1244 pilin trisaccharide α-5NβOHC47NFmPse-(2→4)-β-Xyl-(1→3)-FucNAc. 28837324 1 66 theme important 201:209 arg1 glycans 224:230 important cell surface glycans 201:230 important cell surface glycans 201:230 Pseudaminic acid (Pse) is a nonulosonic acid unique to bacterial species, found as a component of important cell surface glycans and glycoproteins in various pathogenic species, such as the critical hospital threat Pseudomonas aeruginosa. 28821021 4 0 theme ion 609:611 arg1 chromatography 613:626 ion chromatography 609:626 ion chromatography 609:626 The monosaccharide composition was analysed by GC-MS and ion chromatography, respectively. 28821021 7 1 theme glycosyl 1038:1045 arg1 residues 1047:1054 glycosyl residues 1038:1054 glycosyl residues of GASP3-3-I 1038:1067 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 1 theme glycosyl 1038:1045 arg1 GASP3-3-I 1059:1067 GASP3-3-I 1059:1067 GASP3-3-I 1059:1067 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 8 2 theme activity 1237:1244 arg1 assay 1246:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 5 3 theme galacturonic 769:780 arg1 rhamnose 691:698 rhamnose 691:698 rhamnose 691:698 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 5 3 theme galacturonic 769:780 arg1 acid 782:785 galacturonic acid 769:785 galacturonic acid 769:785 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 5 4 theme molar 794:798 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 0 5 from characterization 11:26 arg1 activity 60:67 α-glucosidase activity 46:67 α-glucosidase activity of acidic polysaccharide from Annona squamosa 46:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 6 6 theme oxidation 871:879 arg1 reaction 881:888 periodate oxidation reaction 861:888 periodate oxidation reaction 861:888 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 5 7 theme glucuronic 749:758 arg1 acid 760:763 glucuronic acid 749:763 glucuronic acid 749:763 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 5 7 theme glucuronic 749:758 arg1 rhamnose 691:698 rhamnose 691:698 rhamnose 691:698 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 6 8 theme periodate 861:869 arg1 reaction 881:888 periodate oxidation reaction 861:888 periodate oxidation reaction 861:888 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 7 9 dep l-rhamnose 1196:1205 arg1 1→2 1191:1193 1→2 1191:1193 1→2 1191:1193 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 1 10 theme grading-alcoholic 255:271 arg1 precipitation 273:285 grading-alcoholic precipitation 255:285 grading-alcoholic precipitation 255:285 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 8 11 theme certain 1280:1286 arg1 inhibition 1288:1297 a certain inhibition 1278:1297 a certain inhibition 1278:1297 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 2 12 theme polysaccharide 333:346 arg1 structure 311:319 The structure 307:319 The structure of purified polysaccharide (GASP3-3-I) 307:358 The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. 28821021 0 13 from activity 60:67 arg1 squamosa 106:113 squamosa 106:113 squamosa 106:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 8 14 theme α-glucosidase 1302:1314 arg1 activity 1316:1323 α-glucosidase activity 1302:1323 α-glucosidase activity 1302:1323 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 1 15 theme fruit 172:176 arg1 pulp 178:181 the fruit pulp 168:181 the fruit pulp of Annona squamosa 168:200 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 1 16 theme precipitation 273:285 arg1 combination 240:250 the combination 236:250 the combination of grading-alcoholic precipitation and Sephadex G-200 236:304 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 7 17 dep d-galactose 1136:1146 arg1 1→3 1119:1121 1→3 1119:1121 1→3 1119:1121 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 17 dep d-galactose 1136:1146 arg1 1,3→6 1129:1133 1,3→6 1129:1133 1,3→6 1129:1133 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 6 18 used used 949:952 arg2 NMR 940:942 NMR 940:942 NMR 940:942 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 6 18 used used 949:952 arg2 reaction 881:888 periodate oxidation reaction 861:888 periodate oxidation reaction 861:888 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 6 18 used used 949:952 arg2 methylation 917:927 methylation 917:927 methylation 917:927 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 6 18 used used 949:952 arg2 reaction 907:914 Smith degrading reaction 891:914 Smith degrading reaction 891:914 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 6 18 used used 949:952 arg2 FT-IR 930:934 FT-IR 930:934 FT-IR 930:934 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 2 19 theme purified 324:331 arg1 GASP3-3-I 349:357 GASP3-3-I 349:357 GASP3-3-I 349:357 The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. 28821021 2 19 theme purified 324:331 arg1 polysaccharide 333:346 purified polysaccharide 324:346 purified polysaccharide (GASP3-3-I) 324:358 The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. 28821021 2 20 theme physicochemical 388:402 arg1 analyses 421:428 the physicochemical and instrumental analyses 384:428 the physicochemical and instrumental analyses 384:428 The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. 28821021 0 21 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization 0:26 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 1 22 theme Annona 186:191 arg1 squamosa 193:200 Annona squamosa 186:200 Annona squamosa 186:200 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 7 23 theme 1→ 1150:1151 arg1 d-xylose 1154:1161 (1→) d-xylose 1149:1161 (1→) d-xylose 1149:1161 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 23 theme 1→ 1150:1151 arg1 d-galactose 1136:1146 (1,3→6) d-galactose 1128:1146 (1,3→6) d-galactose 1128:1146 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 1 24 theme Sephadex 291:298 arg1 G-200 300:304 Sephadex G-200 291:304 Sephadex G-200 291:304 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 8 25 contain had 1274:1276 arg1 GASP3-3-I 1264:1272 GASP3-3-I 1264:1272 GASP3-3-I 1264:1272 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 8 25 contain had 1274:1276 arg2 inhibition 1288:1297 a certain inhibition 1278:1297 a certain inhibition 1278:1297 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 5 26 with acid 760:763 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 1 27 theme squamosa 193:200 arg1 pulp 178:181 the fruit pulp 168:181 the fruit pulp of Annona squamosa 168:200 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 1 28 theme G-200 300:304 arg1 combination 240:250 the combination 236:250 the combination of grading-alcoholic precipitation and Sephadex G-200 236:304 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 1 29 theme crude 120:124 arg1 TASP3 142:146 TASP3 142:146 TASP3 142:146 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 1 29 theme crude 120:124 arg1 polysaccharide 126:139 The crude polysaccharide 116:139 The crude polysaccharide (TASP3) 116:147 The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. 28821021 0 30 theme α-glucosidase 46:58 arg1 activity 60:67 α-glucosidase activity 46:67 α-glucosidase activity of acidic polysaccharide from Annona squamosa 46:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 8 31 theme α-glucosidase 1212:1224 arg1 assay 1246:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 5 32 with glucose 729:735 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 2 33 theme instrumental 408:419 arg1 analyses 421:428 the physicochemical and instrumental analyses 384:428 the physicochemical and instrumental analyses 384:428 The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. 28821021 6 34 theme GASP3-3-I 1000:1008 arg1 characterization 980:995 the structural characterization 965:995 the structural characterization of GASP3-3-I 965:1008 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 7 35 dep d-glucose 1179:1187 arg1 3→6 1174:1176 3→6 1174:1176 3→6 1174:1176 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 3 36 theme acidic 475:480 arg1 GASP3-3-I 458:466 GASP3-3-I 458:466 GASP3-3-I 458:466 The results indicated that GASP3-3-I was an acidic heteropolysaccharide and its average molecular weight was 2.28×106Da. 28821021 3 36 theme acidic 475:480 arg1 heteropolysaccharide 482:501 an acidic heteropolysaccharide 472:501 an acidic heteropolysaccharide 472:501 The results indicated that GASP3-3-I was an acidic heteropolysaccharide and its average molecular weight was 2.28×106Da. 28821021 5 37 theme 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 809:848 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 4 38 theme monosaccharide 556:569 arg1 composition 571:581 The monosaccharide composition 552:581 The monosaccharide composition 552:581 The monosaccharide composition was analysed by GC-MS and ion chromatography, respectively. 28821021 7 39 theme 1→ 1094:1095 arg1 d-galactose 1136:1146 (1,3→6) d-galactose 1128:1146 (1,3→6) d-galactose 1128:1146 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 39 theme 1→ 1094:1095 arg1 d-glucose 1179:1187 (3→6) d-glucose 1173:1187 (3→6) d-glucose 1173:1187 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 39 theme 1→ 1094:1095 arg1 l-rhamnose 1196:1205 (1→2) l-rhamnose 1190:1205 (1→2) l-rhamnose 1190:1205 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 39 theme 1→ 1094:1095 arg1 l-arabinose 1098:1108 (1→) l-arabinose 1093:1108 (1→) l-arabinose, (1→6) 1093:1115 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 39 theme 1→ 1094:1095 arg1 1→6 1112:1114 1→6 1112:1114 1→6 1112:1114 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 3 40 theme molecular 519:527 arg1 weight 529:534 its average molecular weight 507:534 its average molecular weight 507:534 The results indicated that GASP3-3-I was an acidic heteropolysaccharide and its average molecular weight was 2.28×106Da. 28821021 6 41 theme degrading 897:905 arg1 reaction 907:914 Smith degrading reaction 891:914 Smith degrading reaction 891:914 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 5 42 with mannose 720:726 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 7 43 theme GASP3-3-I 1059:1067 arg1 residues 1047:1054 glycosyl residues 1038:1054 glycosyl residues of GASP3-3-I 1038:1067 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 7 43 theme GASP3-3-I 1059:1067 arg1 GASP3-3-I 1059:1067 GASP3-3-I 1059:1067 GASP3-3-I 1059:1067 The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. 28821021 8 44 theme inhibitory 1226:1235 arg1 assay 1246:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay 1208:1250 The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity. 28821021 6 45 theme structural 969:978 arg1 characterization 980:995 the structural characterization 965:995 the structural characterization of GASP3-3-I 965:1008 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 6 46 theme Smith 891:895 arg1 reaction 907:914 Smith degrading reaction 891:914 Smith degrading reaction 891:914 Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. 28821021 3 47 theme average 511:517 arg1 weight 529:534 its average molecular weight 507:534 its average molecular weight 507:534 The results indicated that GASP3-3-I was an acidic heteropolysaccharide and its average molecular weight was 2.28×106Da. 28821021 0 48 theme polysaccharide 79:92 arg1 activity 60:67 α-glucosidase activity 46:67 α-glucosidase activity of acidic polysaccharide from Annona squamosa 46:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 0 49 from squamosa 106:113 arg1 activity 60:67 α-glucosidase activity 46:67 α-glucosidase activity of acidic polysaccharide from Annona squamosa 46:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 0 49 from squamosa 106:113 arg1 polysaccharide 79:92 acidic polysaccharide 72:92 acidic polysaccharide from Annona squamosa 72:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 5 50 with acid 782:785 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 5 51 with galactose 738:746 arg1 ratio 800:804 a molar ratio 792:804 a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19 792:848 It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. 28821021 0 52 theme acidic 72:77 arg1 polysaccharide 79:92 acidic polysaccharide 72:92 acidic polysaccharide from Annona squamosa 72:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 28821021 0 53 from inhibition 32:41 arg1 activity 60:67 α-glucosidase activity 46:67 α-glucosidase activity of acidic polysaccharide from Annona squamosa 46:113 Structural characterization and inhibition on α-glucosidase activity of acidic polysaccharide from Annona squamosa. 26182462 0 0 theme N-glycan 91:98 arg1 enrichment 100:109 N-glycan enrichment 91:109 N-glycan enrichment 91:109 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 6 1 theme surface-initiated 967:983 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 10 2 theme high 1548:1551 arg1 efficiency 1564:1573 high enrichment efficiency 1548:1573 high enrichment efficiency 1548:1573 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 7 3 theme strong 1047:1052 arg1 hydrophilicity 1054:1067 The strong hydrophilicity 1043:1067 The strong hydrophilicity of the material 1043:1083 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 1 4 theme important 156:164 arg1 modifications 185:197 the most important post-translational modifications 147:197 the most important post-translational modifications closely correlated with many important biological and pathological processes 147:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 6 5 theme silica 939:944 arg1 microparticles 946:959 silica microparticles 939:959 silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique 939:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 8 6 theme chicken 1213:1219 arg1 albumin 1225:1231 chicken egg albumin 1213:1231 chicken egg albumin 1213:1231 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 6 7 theme in 887:888 arg1 growth 895:900 in situ growth 887:900 in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique 887:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 1 8 theme post-translational 166:183 arg1 modifications 185:197 the most important post-translational modifications 147:197 the most important post-translational modifications closely correlated with many important biological and pathological processes 147:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 4 9 theme N-glycans 619:627 arg1 heterogeneity 598:610 the heterogeneity 594:610 the heterogeneity of the N-glycans 594:627 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 4 9 theme N-glycans 619:627 arg1 abundance 557:565 the low abundance 549:565 the low abundance of the N-glycoproteins 549:588 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 2 10 gly glycoproteins 327:339 arg1 glycoproteins 327:339 glycoproteins 327:339 glycoproteins 327:339 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 0 11 from [Synthesis 0:9 arg1 enrichment 100:109 N-glycan enrichment 91:109 N-glycan enrichment 91:109 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 2 12 link N-linked 307:314 arg1 glycans 316:322 N-linked glycans 307:322 N-linked glycans 307:322 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 4 13 theme protein 664:670 arg1 glycosylation 672:684 the protein glycosylation 660:684 the protein glycosylation 660:684 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 9 14 from proteins 1443:1450 arg1 enrichment 1394:1403 the enrichment 1390:1403 the enrichment of N-linked glycans from human plasma proteins 1390:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 9 14 from proteins 1443:1450 arg1 glycans 1417:1423 N-linked glycans 1408:1423 N-linked glycans from human plasma proteins 1408:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 8 15 used used 1238:1241 arg2 N-glycans 1198:1206 the N-glycans 1194:1206 the N-glycans from chicken egg albumin 1194:1231 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 15 used used 1238:1241 arg2 maltoheptaose 1176:1188 maltoheptaose 1176:1188 maltoheptaose 1176:1188 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 15 used used 1238:1241 arg2 samples 1255:1261 standard samples 1246:1261 standard samples 1246:1261 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 3 16 theme high-throughput 504:518 arg1 profiling 529:537 high-throughput N-glycan profiling 504:537 high-throughput N-glycan profiling 504:537 Therefore, it is very important to establish sensitive methods for high-throughput N-glycan profiling. 26182462 1 17 theme modifications 185:197 arg1 one 140:142 one 140:142 one 140:142 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 1 17 theme modifications 185:197 arg1 modifications 185:197 the most important post-translational modifications 147:197 the most important post-translational modifications closely correlated with many important biological and pathological processes 147:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 5 18 theme protein 832:838 arg1 N-glycans 840:848 protein N-glycans 832:848 protein N-glycans 832:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 10 19 theme application 1591:1601 arg1 value 1603:1607 significant application value 1579:1607 significant application value 1579:1607 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 2 20 theme glycans 316:322 arg1 alterations 292:302 The structural alterations 277:302 The structural alterations of N-linked glycans in glycoproteins 277:339 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 6 21 theme radical 999:1005 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 9 22 from enrichment 1394:1403 arg1 proteins 1443:1450 human plasma proteins 1430:1450 human plasma proteins 1430:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 9 23 theme human 1430:1434 arg1 proteins 1443:1450 human plasma proteins 1430:1450 human plasma proteins 1430:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 7 24 theme complex 1149:1155 arg1 samples 1157:1163 complex samples 1149:1163 complex samples 1149:1163 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 9 25 theme N-linked 1408:1415 arg1 glycans 1417:1423 N-linked glycans 1408:1423 N-linked glycans from human plasma proteins 1408:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 2 26 theme many 368:371 arg1 tumors 429:434 malignant tumors 419:434 malignant tumors 419:434 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 2 26 theme many 368:371 arg1 diseases 373:380 many diseases 368:380 many diseases 368:380 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 2 26 theme many 368:371 arg1 diabetes 391:398 diabetes 391:398 diabetes 391:398 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 2 26 theme many 368:371 arg1 failure 407:413 heart failure 401:413 heart failure 401:413 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 10 27 dep efficiency 1564:1573 arg1 the 1544:1546 the 1544:1546 the 1544:1546 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 2 28 theme malignant 419:427 arg1 tumors 429:434 malignant tumors 419:434 malignant tumors 419:434 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 8 29 theme standard 1246:1253 arg1 maltoheptaose 1176:1188 maltoheptaose 1176:1188 maltoheptaose 1176:1188 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 29 theme standard 1246:1253 arg1 N-glycans 1198:1206 the N-glycans 1194:1206 the N-glycans from chicken egg albumin 1194:1231 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 29 theme standard 1246:1253 arg1 samples 1255:1261 standard samples 1246:1261 standard samples 1246:1261 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 6 30 dep in 887:888 arg1 situ 890:893 situ 890:893 situ 890:893 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 10 31 theme enrichment 1553:1562 arg1 efficiency 1564:1573 high enrichment efficiency 1548:1573 high enrichment efficiency 1548:1573 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 8 32 theme egg 1221:1223 arg1 albumin 1225:1231 chicken egg albumin 1213:1231 chicken egg albumin 1213:1231 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 10 33 theme significant 1579:1589 arg1 value 1603:1607 significant application value 1579:1607 significant application value 1579:1607 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 2 34 from alterations 292:302 arg1 glycoproteins 327:339 glycoproteins 327:339 glycoproteins 327:339 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 6 35 theme polymer 913:919 arg1 growth 895:900 in situ growth 887:900 in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique 887:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 0 36 theme hydrophilic 25:35 arg1 material 59:66 core-shell hydrophilic polymer-silica hybrid material 14:66 core-shell hydrophilic polymer-silica hybrid material 14:66 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 9 37 theme glycans 1417:1423 arg1 enrichment 1394:1403 the enrichment 1390:1403 the enrichment of N-linked glycans from human plasma proteins 1390:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 5 38 theme hydrophilic 743:753 arg1 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 38 theme hydrophilic 743:753 arg1 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 8 39 theme enrichment 1279:1288 arg1 conditions 1290:1299 the enrichment conditions 1275:1299 the enrichment conditions 1275:1299 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 7 40 attach released 1135:1142 arg2 N-glycans 1125:1133 N-glycans 1125:1133 N-glycans released from complex samples 1125:1163 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 7 40 attach released 1135:1142 arg1 samples 1157:1163 complex samples 1149:1163 complex samples 1149:1163 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 1 41 theme many 223:226 arg1 processes 266:274 many important biological and pathological processes 223:274 many important biological and pathological processes 223:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 0 42 theme core-shell 14:23 arg1 material 59:66 core-shell hydrophilic polymer-silica hybrid material 14:66 core-shell hydrophilic polymer-silica hybrid material 14:66 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 5 43 theme polymer-silica 755:768 arg1 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 43 theme polymer-silica 755:768 arg1 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 1 44 theme Protein 113:119 arg1 N-glycosylation 121:135 Protein N-glycosylation 113:135 Protein N-glycosylation 113:135 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 1 45 theme important 228:236 arg1 processes 266:274 many important biological and pathological processes 223:274 many important biological and pathological processes 223:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 0 46 theme hybrid 52:57 arg1 material 59:66 core-shell hydrophilic polymer-silica hybrid material 14:66 core-shell hydrophilic polymer-silica hybrid material 14:66 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 6 47 theme SI-ATRP 1023:1029 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 5 48 theme hybrid 770:775 arg1 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 48 theme hybrid 770:775 arg1 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 3 49 theme sensitive 482:490 arg1 methods 492:498 sensitive methods 482:498 sensitive methods for high-throughput N-glycan profiling 482:537 Therefore, it is very important to establish sensitive methods for high-throughput N-glycan profiling. 26182462 8 50 theme enrichment 1318:1327 arg1 efficiency 1329:1338 the enrichment efficiency 1314:1338 the enrichment efficiency of pGMAG-SiO2 1314:1352 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 1 51 theme biological 238:247 arg1 processes 266:274 many important biological and pathological processes 223:274 many important biological and pathological processes 223:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 0 52 theme polymer-silica 37:50 arg1 material 59:66 core-shell hydrophilic polymer-silica hybrid material 14:66 core-shell hydrophilic polymer-silica hybrid material 14:66 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 9 53 theme plasma 1436:1441 arg1 proteins 1443:1450 human plasma proteins 1430:1450 human plasma proteins 1430:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 5 54 theme N-glycans 840:848 arg1 enrichment 818:827 the efficient enrichment 804:827 the efficient enrichment of protein N-glycans 804:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 0 55 from application 76:86 arg1 enrichment 100:109 N-glycan enrichment 91:109 N-glycan enrichment 91:109 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 6 56 theme transfer 990:997 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 6 57 from growth 895:900 arg1 surface 928:934 the surface 924:934 the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique 924:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 6 58 theme polymerization 1007:1020 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 4 59 gly heterogeneity 598:610 arg1 N-glycoproteins 574:588 the N-glycoproteins 570:588 the N-glycoproteins 570:588 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 4 59 gly heterogeneity 598:610 arg1 N-glycans 619:627 the N-glycans 615:627 the N-glycans 615:627 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 7 60 theme N-glycans 1125:1133 arg1 enrichment 1111:1120 the enrichment 1107:1120 the enrichment of N-glycans released from complex samples 1107:1163 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 0 61 theme material 59:66 arg1 [Synthesis 0:9 [Synthesis 0:9 [Synthesis of core-shell hydrophilic polymer-silica hybrid material 0:66 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 0 61 theme material 59:66 arg1 application 76:86 its application 72:86 its application in N-glycan enrichment 72:109 [Synthesis of core-shell hydrophilic polymer-silica hybrid material and its application in N-glycan enrichment]. 26182462 5 62 contain had 716:718 arg2 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 62 contain had 716:718 arg1 we 713:714 we 713:714 we 713:714 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 62 contain had 716:718 arg2 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 10 63 theme pGMAG-SiO2 1612:1621 arg1 efficiency 1564:1573 high enrichment efficiency 1548:1573 high enrichment efficiency 1548:1573 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 10 63 theme pGMAG-SiO2 1612:1621 arg1 value 1603:1607 significant application value 1579:1607 significant application value 1579:1607 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 6 64 theme glucose 905:911 arg1 polymer 913:919 glucose polymer 905:919 glucose polymer 905:919 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 4 65 theme N-glycoproteins 574:588 arg1 heterogeneity 598:610 the heterogeneity 594:610 the heterogeneity of the N-glycans 594:627 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 4 65 theme N-glycoproteins 574:588 arg1 abundance 557:565 the low abundance 549:565 the low abundance of the N-glycoproteins 549:588 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 5 66 theme synthesized 720:730 arg1 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 66 theme synthesized 720:730 arg1 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 2 67 theme N-linked 307:314 arg1 glycans 316:322 N-linked glycans 307:322 N-linked glycans 307:322 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 7 68 theme material 1076:1083 arg1 hydrophilicity 1054:1067 The strong hydrophilicity 1043:1067 The strong hydrophilicity of the material 1043:1083 The strong hydrophilicity of the material makes it suitable for the enrichment of N-glycans released from complex samples. 26182462 3 69 theme N-glycan 520:527 arg1 profiling 529:537 high-throughput N-glycan profiling 504:537 high-throughput N-glycan profiling 504:537 Therefore, it is very important to establish sensitive methods for high-throughput N-glycan profiling. 26182462 9 70 link N-linked 1408:1415 arg1 glycans 1417:1423 N-linked glycans 1408:1423 N-linked glycans from human plasma proteins 1408:1450 Finally, pGMAG-SiO2 was applied to the enrichment of N-linked glycans from human plasma proteins and 47 glycoforms were successfully identified after enrichment. 26182462 1 71 theme pathological 253:264 arg1 processes 266:274 many important biological and pathological processes 223:274 many important biological and pathological processes 223:274 Protein N-glycosylation is one of the most important post-translational modifications closely correlated with many important biological and pathological processes. 26182462 5 72 theme core-shell 732:741 arg1 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 pGMAG-SiO2 788:797 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 5 72 theme core-shell 732:741 arg1 materials 777:785 synthesized core-shell hydrophilic polymer-silica hybrid materials 720:785 synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans 720:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 4 73 theme low 553:555 arg1 abundance 557:565 the low abundance 549:565 the low abundance of the N-glycoproteins 549:588 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 8 74 from albumin 1225:1231 arg1 maltoheptaose 1176:1188 maltoheptaose 1176:1188 maltoheptaose 1176:1188 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 74 from albumin 1225:1231 arg1 N-glycans 1198:1206 the N-glycans 1194:1206 the N-glycans from chicken egg albumin 1194:1231 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 74 from albumin 1225:1231 arg1 samples 1255:1261 standard samples 1246:1261 standard samples 1246:1261 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 8 75 theme pGMAG-SiO2 1343:1352 arg1 efficiency 1329:1338 the enrichment efficiency 1314:1338 the enrichment efficiency of pGMAG-SiO2 1314:1352 Secondly, maltoheptaose and the N-glycans from chicken egg albumin were used as standard samples to optimize the enrichment conditions and evaluate the enrichment efficiency of pGMAG-SiO2. 26182462 10 76 theme N-glycans 1642:1650 arg1 analysis 1630:1637 the analysis 1626:1637 the analysis of N-glycans 1626:1650 These results demonstrated the high enrichment efficiency and significant application value of pGMAG-SiO2 in the analysis of N-glycans. 26182462 6 77 theme microparticles 946:959 arg1 surface 928:934 the surface 924:934 the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique 924:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 5 78 theme efficient 808:816 arg1 enrichment 818:827 the efficient enrichment 804:827 the efficient enrichment of protein N-glycans 804:848 In this work, we had synthesized core-shell hydrophilic polymer-silica hybrid materials (pGMAG-SiO2) for the efficient enrichment of protein N-glycans. 26182462 6 79 theme atom 985:988 arg1 technique 1032:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 surface-initiated atom transfer radical polymerization (SI-ATRP) technique 967:1040 Firstly, pGMAG-SiO2 was prepared by in situ growth of glucose polymer on the surface of silica microparticles using surface-initiated atom transfer radical polymerization (SI-ATRP) technique. 26182462 2 80 theme structural 281:290 arg1 alterations 292:302 The structural alterations 277:302 The structural alterations of N-linked glycans in glycoproteins 277:339 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26182462 4 81 gly N-glycoproteins 574:588 arg1 N-glycoproteins 574:588 the N-glycoproteins 570:588 the N-glycoproteins 570:588 However, the low abundance of the N-glycoproteins and the heterogeneity of the N-glycans make it a challenge to analyse the protein glycosylation sensitively. 26182462 2 82 theme heart 401:405 arg1 failure 407:413 heart failure 401:413 heart failure 401:413 The structural alterations of N-linked glycans in glycoproteins are always associated with many diseases, such as diabetes, heart failure and malignant tumors. 26139606 8 0 theme maltodextrin 1541:1552 arg1 products 1554:1561 maltodextrin products 1541:1561 maltodextrin products 1541:1561 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 1 1 from metabolism 144:153 arg1 coli 199:202 Escherichia coli 187:202 Escherichia coli 187:202 Amylomaltase MalQ is essential for the metabolism of maltose and maltodextrins in Escherichia coli. 26139606 3 2 dep analog 586:591 arg1 acarviosine-glucose-acarbose 593:620 acarviosine-glucose-acarbose 593:620 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 9 3 theme structural 1691:1700 arg1 data 1716:1719 both structural and enzymatic data 1686:1719 both structural and enzymatic data 1686:1719 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 8 4 theme HPLC-based 1452:1461 arg1 assay 1475:1479 an HPLC-based MalQ enzyme assay 1449:1479 an HPLC-based MalQ enzyme assay 1449:1479 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 3 5 dep apo-form 499:506 arg1 i.e. 490:493 i.e. 490:493 i.e. 490:493 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 2 6 theme linear 325:330 arg1 maltodextrins 332:344 linear maltodextrins 325:344 linear maltodextrins of various lengths 325:363 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 4 7 with contrast 810:817 arg1 enzymes 858:864 previously described thermophilic enzymes 824:864 previously described thermophilic enzymes 824:864 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 3 8 theme state 580:584 arg1 analog 586:591 the transition state analog 565:591 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 5 9 theme catalytic 1015:1023 arg1 core 1025:1028 the catalytic core 1011:1028 the catalytic core 1011:1028 This moiety seems unique to amylomaltases from Enterobacteriaceae and folds into two distinct subdomains that associate with different parts of the catalytic core. 26139606 4 10 theme amylomaltase 713:724 arg1 example 679:685 the first example 669:685 the first example of a mesophilic bacterial amylomaltase with known structure 669:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 4 11 with amylomaltase 713:724 arg1 structure 737:745 known structure 731:745 known structure 731:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 9 12 theme enzymatic 1706:1714 arg1 data 1716:1719 both structural and enzymatic data 1686:1719 both structural and enzymatic data 1686:1719 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 4 13 theme N-terminal 763:772 arg1 extension 774:782 an N-terminal extension 760:782 an N-terminal extension 760:782 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 5 14 theme core 1025:1028 arg1 parts 1002:1006 different parts 992:1006 different parts of the catalytic core 992:1028 This moiety seems unique to amylomaltases from Enterobacteriaceae and folds into two distinct subdomains that associate with different parts of the catalytic core. 26139606 1 15 theme maltodextrins 170:182 arg1 metabolism 144:153 the metabolism 140:153 the metabolism of maltose and maltodextrins in Escherichia coli 140:202 Amylomaltase MalQ is essential for the metabolism of maltose and maltodextrins in Escherichia coli. 26139606 1 16 theme maltose 158:164 arg1 metabolism 144:153 the metabolism 140:153 the metabolism of maltose and maltodextrins in Escherichia coli 140:202 Amylomaltase MalQ is essential for the metabolism of maltose and maltodextrins in Escherichia coli. 26139606 0 17 theme coli 99:102 arg1 Amylomaltase 71:82 the Amylomaltase 67:82 the Amylomaltase of Escherichia coli 67:102 Structural Basis for the Interconversion of Maltodextrins by MalQ, the Amylomaltase of Escherichia coli. 26139606 0 17 theme coli 99:102 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis for the Interconversion of Maltodextrins by MalQ 0:64 Structural Basis for the Interconversion of Maltodextrins by MalQ, the Amylomaltase of Escherichia coli. 26139606 7 18 theme acceptor 1344:1351 arg1 subsite 1353:1359 the acceptor subsite +1 1340:1362 the acceptor subsite +1 1340:1362 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 3 19 theme inhibitor 542:550 arg1 complex 552:558 an inhibitor complex 539:558 an inhibitor complex with the transition state analog acarviosine-glucose-acarbose 539:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 4 20 theme first 673:677 arg1 example 679:685 the first example 669:685 the first example of a mesophilic bacterial amylomaltase with known structure 669:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 7 21 theme inhibitor 1228:1236 arg1 complex 1238:1244 the inhibitor complex 1224:1244 the inhibitor complex 1224:1244 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 9 22 from metabolism 1848:1857 arg1 coli 1882:1885 E. coli 1879:1885 E. coli 1879:1885 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 4 23 from extension 774:782 arg1 contrast 810:817 contrast 810:817 contrast with previously described thermophilic enzymes 810:864 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 2 24 theme various 349:355 arg1 lengths 357:363 various lengths 349:363 various lengths 349:363 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 9 25 from maltodextrins 1862:1874 arg1 coli 1882:1885 E. coli 1879:1885 E. coli 1879:1885 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 3 26 theme crystal 455:461 arg1 structures 463:472 three crystal structures 449:472 three crystal structures 449:472 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 0 27 theme Structural 0:9 arg1 Amylomaltase 71:82 the Amylomaltase 67:82 the Amylomaltase of Escherichia coli 67:102 Structural Basis for the Interconversion of Maltodextrins by MalQ, the Amylomaltase of Escherichia coli. 26139606 0 27 theme Structural 0:9 arg1 Basis 11:15 Structural Basis 0:15 Structural Basis for the Interconversion of Maltodextrins by MalQ 0:64 Structural Basis for the Interconversion of Maltodextrins by MalQ, the Amylomaltase of Escherichia coli. 26139606 8 28 theme substrate 1600:1608 arg1 length 1578:1583 the length 1574:1583 the length of the initial substrate 1574:1608 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 3 29 with complex 513:519 arg1 maltose 526:532 maltose 526:532 maltose 526:532 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 29 with complex 513:519 arg1 analog 586:591 the transition state analog 565:591 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 5 30 theme different 992:1000 arg1 parts 1002:1006 different parts 992:1006 different parts of the catalytic core 992:1028 This moiety seems unique to amylomaltases from Enterobacteriaceae and folds into two distinct subdomains that associate with different parts of the catalytic core. 26139606 3 31 theme transglycosylation 402:419 arg1 basis 393:397 the molecular basis 379:397 the molecular basis of transglycosylation 379:419 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 32 theme down 638:641 arg1 resolutions 626:636 resolutions 626:636 resolutions down to 2.1 Å 626:650 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 7 33 theme catalyst 1425:1432 arg1 orientation 1396:1406 the catalytically competent orientation 1368:1406 the catalytically competent orientation of the acid/base catalyst Glu-496 1368:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 6 34 theme distinct 1103:1110 arg1 states 1112:1117 distinct states 1103:1117 distinct states of this enzyme 1103:1132 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 9 35 theme maltodextrins 1862:1874 arg1 metabolism 1848:1857 the metabolism 1844:1857 the metabolism of maltodextrins in E. coli 1844:1885 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 3 36 theme transition 569:578 arg1 analog 586:591 the transition state analog 565:591 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 6 37 theme conformational 1158:1171 arg1 changes 1173:1179 considerable conformational changes 1145:1179 considerable conformational changes 1145:1179 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 9 38 with consistent 1725:1734 arg1 rates 1770:1774 the extremely low hydrolysis rates 1741:1774 the extremely low hydrolysis rates observed for amylomaltases 1741:1801 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 7 39 theme 4-OH 1300:1303 arg1 group 1305:1309 a 4-OH group 1298:1309 a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 1298:1362 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 39 theme 4-OH 1300:1303 arg1 bond 1331:1334 α1-4-glycosidic bond 1315:1334 α1-4-glycosidic bond 1315:1334 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 4 40 theme thermophilic 845:856 arg1 enzymes 858:864 previously described thermophilic enzymes 824:864 previously described thermophilic enzymes 824:864 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 3 41 theme enzyme 482:487 arg1 complex 552:558 an inhibitor complex 539:558 an inhibitor complex with the transition state analog acarviosine-glucose-acarbose 539:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 41 theme enzyme 482:487 arg1 complex 513:519 its complex 509:519 its complex with maltose 509:532 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 41 theme enzyme 482:487 arg1 structures 463:472 three crystal structures 449:472 three crystal structures 449:472 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 6 42 theme enzyme 1127:1132 arg1 states 1112:1117 distinct states 1103:1117 distinct states of this enzyme 1103:1132 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 9 43 theme low 1755:1757 arg1 rates 1770:1774 the extremely low hydrolysis rates 1741:1774 the extremely low hydrolysis rates observed for amylomaltases 1741:1801 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 7 44 from requirement 1261:1271 arg1 subsite 1353:1359 the acceptor subsite +1 1340:1362 the acceptor subsite +1 1340:1362 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 4 45 theme described 835:843 arg1 enzymes 858:864 previously described thermophilic enzymes 824:864 previously described thermophilic enzymes 824:864 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 9 46 theme MalQ 1835:1838 arg1 importance 1821:1830 the importance 1817:1830 the importance of MalQ for the metabolism of maltodextrins in E. coli 1817:1885 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 3 47 theme molecular 383:391 arg1 basis 393:397 the molecular basis 379:397 the molecular basis of transglycosylation 379:419 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 48 with complex 552:558 arg1 maltose 526:532 maltose 526:532 maltose 526:532 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 48 with complex 552:558 arg1 analog 586:591 the transition state analog 565:591 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 8 49 theme increasing 1616:1625 arg1 numbers 1627:1633 increasing numbers 1616:1633 increasing numbers of glycosidic bonds 1616:1653 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 9 50 theme hydrolysis 1759:1768 arg1 rates 1770:1774 the extremely low hydrolysis rates 1741:1774 the extremely low hydrolysis rates observed for amylomaltases 1741:1801 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 8 51 theme initial 1592:1598 arg1 substrate 1600:1608 the initial substrate 1588:1608 the initial substrate 1588:1608 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 2 52 theme transglycosylation/disproportionation 218:254 arg1 reactions 256:264 transglycosylation/disproportionation reactions 218:264 transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths 218:363 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 1 53 theme Amylomaltase 105:116 arg1 MalQ 118:121 Amylomaltase MalQ 105:121 Amylomaltase MalQ 105:121 Amylomaltase MalQ is essential for the metabolism of maltose and maltodextrins in Escherichia coli. 26139606 0 54 theme Maltodextrins 44:56 arg1 Interconversion 25:39 the Interconversion 21:39 the Interconversion of Maltodextrins by MalQ 21:64 Structural Basis for the Interconversion of Maltodextrins by MalQ, the Amylomaltase of Escherichia coli. 26139606 8 55 theme glycosidic 1638:1647 arg1 bonds 1649:1653 glycosidic bonds 1638:1653 glycosidic bonds 1638:1653 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 3 56 with structures 463:472 arg1 maltose 526:532 maltose 526:532 maltose 526:532 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 3 56 with structures 463:472 arg1 analog 586:591 the transition state analog 565:591 the transition state analog acarviosine-glucose-acarbose 565:620 To elucidate the molecular basis of transglycosylation by MalQ, we have determined three crystal structures of this enzyme, i.e. the apo-form, its complex with maltose, and an inhibitor complex with the transition state analog acarviosine-glucose-acarbose, at resolutions down to 2.1 Å. 26139606 8 57 theme equilibrium 1512:1522 arg1 concentration 1524:1536 the equilibrium concentration 1508:1536 the equilibrium concentration of maltodextrin products 1508:1561 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 7 58 theme acid/base 1415:1423 arg1 catalyst 1425:1432 the acid/base catalyst Glu-496 1411:1440 the acid/base catalyst Glu-496 1411:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 59 theme α1-4-glycosidic 1315:1329 arg1 group 1305:1309 a 4-OH group 1298:1309 a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 1298:1362 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 59 theme α1-4-glycosidic 1315:1329 arg1 bond 1331:1334 α1-4-glycosidic bond 1315:1334 α1-4-glycosidic bond 1315:1334 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 4 60 theme known 731:735 arg1 structure 737:745 known structure 731:745 known structure 731:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 7 61 theme group 1305:1309 arg1 requirement 1261:1271 the requirement 1257:1271 the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496 1257:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 8 62 theme enzyme 1468:1473 arg1 assay 1475:1479 an HPLC-based MalQ enzyme assay 1449:1479 an HPLC-based MalQ enzyme assay 1449:1479 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 2 63 theme dextrinyl 287:295 arg1 units 297:301 glycosyl or dextrinyl units 275:301 units 297:301 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 9 64 from coli 1882:1885 arg1 metabolism 1848:1857 the metabolism 1844:1857 the metabolism of maltodextrins in E. coli 1844:1885 Thus, both structural and enzymatic data are consistent with the extremely low hydrolysis rates observed for amylomaltases and underline the importance of MalQ for the metabolism of maltodextrins in E. coli. 26139606 4 65 theme residues 797:804 arg1 extension 774:782 an N-terminal extension 760:782 an N-terminal extension 760:782 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 5 66 theme distinct 952:959 arg1 subdomains 961:970 two distinct subdomains 948:970 two distinct subdomains that associate with different parts of the catalytic core 948:1028 This moiety seems unique to amylomaltases from Enterobacteriaceae and folds into two distinct subdomains that associate with different parts of the catalytic core. 26139606 8 67 theme MalQ 1463:1466 arg1 assay 1475:1479 an HPLC-based MalQ enzyme assay 1449:1479 an HPLC-based MalQ enzyme assay 1449:1479 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 7 68 from subsite 1353:1359 arg1 requirement 1261:1271 the requirement 1257:1271 the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496 1257:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 68 from subsite 1353:1359 arg1 group 1305:1309 a 4-OH group 1298:1309 a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 1298:1362 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 68 from subsite 1353:1359 arg1 bond 1331:1334 α1-4-glycosidic bond 1315:1334 α1-4-glycosidic bond 1315:1334 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 7 68 from subsite 1353:1359 arg1 group 1288:1292 a 3-OH group 1281:1292 a 3-OH group 1281:1292 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 2 69 theme lengths 357:363 arg1 maltodextrins 332:344 linear maltodextrins 325:344 linear maltodextrins of various lengths 325:363 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 6 70 theme catalytic 1192:1200 arg1 cycle 1202:1206 the catalytic cycle 1188:1206 the catalytic cycle 1188:1206 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 8 71 theme bonds 1649:1653 arg1 numbers 1627:1633 increasing numbers 1616:1633 increasing numbers of glycosidic bonds 1616:1653 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 7 72 theme 3-OH 1283:1286 arg1 group 1288:1292 a 3-OH group 1281:1292 a 3-OH group 1281:1292 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 6 73 theme considerable 1145:1156 arg1 changes 1173:1179 considerable conformational changes 1145:1179 considerable conformational changes 1145:1179 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 2 74 theme glycosyl 275:282 arg1 units 297:301 glycosyl or dextrinyl units 275:301 units 297:301 It catalyzes transglycosylation/disproportionation reactions in which glycosyl or dextrinyl units are transferred among linear maltodextrins of various lengths. 26139606 5 75 from Enterobacteriaceae 914:931 arg1 amylomaltases 895:907 amylomaltases 895:907 amylomaltases from Enterobacteriaceae 895:931 This moiety seems unique to amylomaltases from Enterobacteriaceae and folds into two distinct subdomains that associate with different parts of the catalytic core. 26139606 4 76 theme bacterial 703:711 arg1 amylomaltase 713:724 a mesophilic bacterial amylomaltase 690:724 a mesophilic bacterial amylomaltase with known structure 690:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 6 77 theme crystal 1060:1066 arg1 structures 1068:1077 the three MalQ crystal structures 1045:1077 the three MalQ crystal structures 1045:1077 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 26139606 7 78 theme group 1288:1292 arg1 requirement 1261:1271 the requirement 1257:1271 the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496 1257:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 8 79 theme products 1554:1561 arg1 concentration 1524:1536 the equilibrium concentration 1508:1536 the equilibrium concentration of maltodextrin products 1508:1561 Using an HPLC-based MalQ enzyme assay, we could demonstrate that the equilibrium concentration of maltodextrin products depends on the length of the initial substrate; with increasing numbers of glycosidic bonds, less glucose is formed. 26139606 7 80 theme competent 1386:1394 arg1 orientation 1396:1406 the catalytically competent orientation 1368:1406 the catalytically competent orientation of the acid/base catalyst Glu-496 1368:1440 In particular, the inhibitor complex highlights the requirement of both a 3-OH group and a 4-OH group (or α1-4-glycosidic bond) at the acceptor subsite +1 for the catalytically competent orientation of the acid/base catalyst Glu-496. 26139606 4 81 theme mesophilic 692:701 arg1 amylomaltase 713:724 a mesophilic bacterial amylomaltase 690:724 a mesophilic bacterial amylomaltase with known structure 690:745 MalQ represents the first example of a mesophilic bacterial amylomaltase with known structure and exhibits an N-terminal extension of about 140 residues, in contrast with previously described thermophilic enzymes. 26139606 6 82 theme MalQ 1055:1058 arg1 structures 1068:1077 the three MalQ crystal structures 1045:1077 the three MalQ crystal structures 1045:1077 Intriguingly, the three MalQ crystal structures appear to correspond to distinct states of this enzyme, revealing considerable conformational changes during the catalytic cycle. 28301166 12 0 theme interactions 2146:2157 arg1 investigations 2110:2123 investigations 2110:2123 investigations of B. hyodysenteriae interactions with the host via mucin glycans 2110:2189 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 10 1 theme mucin 1796:1800 arg1 glycosylation 1802:1814 colonic mucin glycosylation 1788:1814 colonic mucin glycosylation in pigs 1788:1822 Therefore, we conclude that B. hyodysenteriae plays a major role in regulating colonic mucin glycosylation in pigs during SD. 28301166 6 2 theme healthy 961:967 arg1 pigs 969:972 healthy pigs 961:972 healthy pigs having higher interindividual variation 961:1012 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 3 attach present 1064:1070 arg2 present 1064:1070 present 1064:1070 present 1064:1070 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 3 attach present 1064:1070 arg2 array 1033:1037 a larger array 1024:1037 a larger array of glycan structures 1024:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 3 attach present 1064:1070 arg1 pigs 1084:1087 infected pigs 1075:1087 infected pigs 1075:1087 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 11 4 theme epitopes 1974:1981 arg1 exposure 1962:1969 increased exposure 1952:1969 increased exposure of epitopes important for host-pathogen interactions 1952:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 7 5 theme specific 1164:1171 arg1 glycans 1191:1197 specific infection-related glycans 1164:1197 specific infection-related glycans 1164:1197 This implied that infection induced loss of individual variation and that specific infection-related glycans were induced. 28301166 0 6 theme Brachyspira 0:10 arg1 Infection 27:35 Brachyspira hyodysenteriae Infection 0:35 Brachyspira hyodysenteriae Infection 0:35 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 8 7 theme noninfected 1277:1287 arg1 pigs 1289:1292 noninfected pigs 1277:1292 noninfected pigs 1277:1292 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 6 8 contain having 974:979 arg1 pigs 969:972 healthy pigs 961:972 healthy pigs having higher interindividual variation 961:1012 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 8 contain having 974:979 arg2 variation 1004:1012 higher interindividual variation 981:1012 higher interindividual variation 981:1012 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 2 9 theme B. 315:316 arg1 sites 341:345 B. hyodysenteriae binding sites 315:345 B. hyodysenteriae binding sites on mucins 315:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 11 10 from changes 1839:1845 arg1 O-glycosylation 1856:1870 mucin O-glycosylation 1850:1870 mucin O-glycosylation 1850:1870 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 8 11 theme increased 1366:1374 arg1 levels 1376:1381 increased levels 1366:1381 increased levels of the C2GnT glycosyl transferase 1366:1415 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 7 12 theme individual 1134:1143 arg1 variation 1145:1153 individual variation 1134:1153 individual variation 1134:1153 This implied that infection induced loss of individual variation and that specific infection-related glycans were induced. 28301166 12 13 with interactions 2146:2157 arg1 host 2168:2171 the host 2164:2171 the host via mucin glycans 2164:2189 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 11 14 theme increased 1952:1960 arg1 exposure 1962:1969 increased exposure 1952:1969 increased exposure of epitopes important for host-pathogen interactions 1952:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 1 15 theme global 215:220 arg1 losses 232:237 global financial losses 215:237 global financial losses 215:237 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 0 16 from Expression 99:108 arg1 Colon 141:145 Porcine Colon 133:145 Porcine Colon 133:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 11 17 theme important 1983:1991 arg1 epitopes 1974:1981 epitopes 1974:1981 epitopes important for host-pathogen interactions 1974:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 8 18 theme glycosyl 1396:1403 arg1 transferase 1405:1415 the C2GnT glycosyl transferase 1386:1415 the C2GnT glycosyl transferase 1386:1415 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 8 19 theme infected 1326:1333 arg1 animals 1335:1341 infected animals 1326:1341 infected animals 1326:1341 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 8 20 theme C2GnT 1390:1394 arg1 transferase 1405:1415 the C2GnT glycosyl transferase 1386:1415 the C2GnT glycosyl transferase 1386:1415 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 11 21 theme glycan 1891:1896 arg1 fingerprint 1898:1908 a glycan fingerprint 1889:1908 a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions 1889:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 6 22 theme interindividual 988:1002 arg1 variation 1004:1012 higher interindividual variation 981:1012 higher interindividual variation 981:1012 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 8 23 theme transferase 1405:1415 arg1 levels 1376:1381 increased levels 1366:1381 increased levels of the C2GnT glycosyl transferase 1366:1415 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 5 24 theme colon 809:813 arg1 tissues 815:821 five healthy colon tissues 796:821 five healthy colon tissues 796:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 1 25 theme swine 182:186 arg1 SD 199:200 SD 199:200 SD 199:200 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 1 25 theme swine 182:186 arg1 dysentery 188:196 swine dysentery 182:196 swine dysentery (SD) 182:201 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 11 26 theme mucin 1850:1854 arg1 O-glycosylation 1856:1870 mucin O-glycosylation 1850:1870 mucin O-glycosylation 1850:1870 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 9 27 theme structures 1621:1630 arg1 abundance 1608:1616 a lower abundance 1600:1616 a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1600:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 0 28 theme Core-2 113:118 arg1 O-Glycans 120:128 Core-2 O-Glycans 113:128 Core-2 O-Glycans 113:128 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 6 29 theme glycan 1042:1047 arg1 structures 1049:1058 glycan structures 1042:1058 glycan structures 1042:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 9 30 from pigs 1455:1458 arg1 chains 1434:1439 glycan chains 1427:1439 glycan chains from infected pigs 1427:1458 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 4 31 theme mucin 574:578 arg1 O-glycome 580:588 the swine colonic mucin O-glycome 556:588 the swine colonic mucin O-glycome 556:588 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 6 32 from present 1064:1070 arg1 pigs 1084:1087 infected pigs 1075:1087 infected pigs 1075:1087 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 0 33 theme Increased 89:97 arg1 Expression 99:108 an Increased Expression 86:108 an Increased Expression of Core-2 O-Glycans in Porcine Colon 86:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 5 34 attach isolated 764:771 arg2 mucins 757:762 soluble and insoluble mucins 735:762 soluble and insoluble mucins isolated from five infected and five healthy colon tissues 735:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 5 34 attach isolated 764:771 arg1 tissues 815:821 five healthy colon tissues 796:821 five healthy colon tissues 796:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 5 34 attach isolated 764:771 arg1 infected 783:790 infected 783:790 infected 783:790 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 2 35 theme mucin 387:391 arg1 secretion 393:401 increased colonic mucin secretion 369:401 increased colonic mucin secretion 369:401 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 5 36 theme liquid 868:873 arg1 chromatography 875:888 porous graphitized carbon liquid chromatography 842:888 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 4 37 theme swine 560:564 arg1 O-glycome 580:588 the swine colonic mucin O-glycome 556:588 the swine colonic mucin O-glycome 556:588 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 5 38 attach released 721:728 arg1 mucins 757:762 soluble and insoluble mucins 735:762 soluble and insoluble mucins isolated from five infected and five healthy colon tissues 735:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 5 38 attach released 721:728 arg2 O-Glycans 695:703 O-Glycans 695:703 O-Glycans 695:703 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 10 39 theme major 1763:1767 arg1 role 1769:1772 a major role 1761:1772 a major role 1761:1772 Therefore, we conclude that B. hyodysenteriae plays a major role in regulating colonic mucin glycosylation in pigs during SD. 28301166 5 40 theme tandem 890:895 arg1 spectrometry 902:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 9 41 theme infected 1446:1453 arg1 pigs 1455:1458 infected pigs 1446:1458 infected pigs 1446:1458 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 9 42 gly fucosylated 1642:1652 arg1 structures 1621:1630 structures 1621:1630 structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1621:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 0 43 theme Mucin 47:51 arg1 Synthesis 67:75 Mucin Glycosylation Synthesis 47:75 Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon 47:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 3 44 theme layer 505:509 arg1 pattern 465:471 the glycosylation pattern 447:471 the glycosylation pattern of the porcine intestinal mucus layer 447:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 5 45 theme graphitized 849:859 arg1 chromatography 875:888 porous graphitized carbon liquid chromatography 842:888 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 8 46 from O-glycans 1264:1272 arg1 pigs 1289:1292 noninfected pigs 1277:1292 noninfected pigs 1277:1292 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 4 47 dep hyodysenteriae-infected 649:671 arg1 B. 646:647 B. hyodysenteriae-infected and noninfected pigs 646:692 B. 646:647 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 12 48 dep B. 2128:2129 arg1 hyodysenteriae 2131:2144 hyodysenteriae 2131:2144 hyodysenteriae 2131:2144 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 3 49 gly glycosylation 451:463 arg1 layer 505:509 the porcine intestinal mucus layer 476:509 the porcine intestinal mucus layer 476:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 3 50 theme intestinal 488:497 arg1 layer 505:509 the porcine intestinal mucus layer 476:509 the porcine intestinal mucus layer 476:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 3 51 theme host 527:530 arg1 niche 532:536 its host niche 523:536 its host niche 523:536 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 1 52 theme pig 246:248 arg1 industry 250:257 the pig industry 242:257 the pig industry 242:257 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 6 53 theme higher 981:986 arg1 variation 1004:1012 higher interindividual variation 981:1012 higher interindividual variation 981:1012 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 9 54 theme structures 1503:1512 arg1 abundance 1490:1498 a higher abundance 1481:1498 a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1481:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 8 55 theme dominating 1217:1226 arg1 structures 1228:1237 The dominating structures 1213:1237 The dominating structures 1213:1237 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 11 56 from fingerprint 1898:1908 arg1 mucus 1929:1933 porcine colonic mucus 1913:1933 porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions 1913:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 2 57 from sites 341:345 arg1 mucins 350:355 mucins 350:355 mucins 350:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 7 58 theme infection-related 1173:1189 arg1 glycans 1191:1197 specific infection-related glycans 1164:1197 specific infection-related glycans 1164:1197 This implied that infection induced loss of individual variation and that specific infection-related glycans were induced. 28301166 2 59 from mucins 350:355 arg1 increase 303:310 an increase 300:310 an increase in B. hyodysenteriae binding sites on mucins 300:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 10 60 theme colonic 1788:1794 arg1 glycosylation 1802:1814 colonic mucin glycosylation 1788:1814 colonic mucin glycosylation in pigs 1788:1822 Therefore, we conclude that B. hyodysenteriae plays a major role in regulating colonic mucin glycosylation in pigs during SD. 28301166 8 61 theme core-4-type 1252:1262 arg1 O-glycans 1264:1272 core-4-type O-glycans 1252:1272 core-4-type O-glycans in noninfected pigs 1252:1292 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 8 62 from O-glycans 1313:1321 arg1 animals 1335:1341 infected animals 1326:1341 infected animals 1326:1341 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 10 63 from glycosylation 1802:1814 arg1 pigs 1819:1822 pigs 1819:1822 pigs 1819:1822 Therefore, we conclude that B. hyodysenteriae plays a major role in regulating colonic mucin glycosylation in pigs during SD. 28301166 7 64 theme variation 1145:1153 arg1 loss 1126:1129 loss 1126:1129 loss of individual variation 1126:1153 This implied that infection induced loss of individual variation and that specific infection-related glycans were induced. 28301166 12 65 theme potential 2061:2069 arg1 targets 2083:2089 potential therapeutic targets 2061:2089 potential therapeutic targets 2061:2089 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 5 66 theme insoluble 747:755 arg1 mucins 757:762 soluble and insoluble mucins 735:762 soluble and insoluble mucins isolated from five infected and five healthy colon tissues 735:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 9 67 theme noninfected 1691:1701 arg1 pigs 1703:1706 noninfected pigs 1691:1706 noninfected pigs 1691:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 5 68 theme mass 897:900 arg1 spectrometry 902:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 6 69 theme larger 1026:1031 arg1 present 1064:1070 present 1064:1070 present 1064:1070 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 69 theme larger 1026:1031 arg1 array 1033:1037 a larger array 1024:1037 a larger array of glycan structures 1024:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 2 70 theme increased 369:377 arg1 secretion 393:401 increased colonic mucin secretion 369:401 increased colonic mucin secretion 369:401 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 1 71 theme financial 222:230 arg1 losses 232:237 global financial losses 215:237 global financial losses 215:237 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 8 72 theme core-2-type 1301:1311 arg1 O-glycans 1313:1321 core-2-type O-glycans 1301:1321 core-2-type O-glycans 1301:1321 The dominating structures shifted from core-4-type O-glycans in noninfected pigs toward core-2-type O-glycans in infected animals, which correlated with increased levels of the C2GnT glycosyl transferase. 28301166 2 73 with Infection 260:268 arg1 pathogen 280:287 this pathogen 275:287 this pathogen 275:287 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 4 74 theme noninfected 677:687 arg1 pigs 689:692 B. hyodysenteriae-infected and noninfected pigs 646:692 pigs 689:692 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 4 75 from differences 609:619 arg1 glycosylation 624:636 glycosylation 624:636 glycosylation 624:636 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 12 76 theme mucin 2177:2181 arg1 glycans 2183:2189 mucin glycans 2177:2189 mucin glycans 2177:2189 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 5 77 theme soluble 735:741 arg1 mucins 757:762 soluble and insoluble mucins 735:762 soluble and insoluble mucins isolated from five infected and five healthy colon tissues 735:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 1 78 theme Brachyspira 148:158 arg1 hyodysenteriae 160:173 Brachyspira hyodysenteriae 148:173 Brachyspira hyodysenteriae 148:173 Brachyspira hyodysenteriae causes swine dysentery (SD), leading to global financial losses to the pig industry. 28301166 2 79 dep B. 315:316 arg1 hyodysenteriae 318:331 hyodysenteriae 318:331 hyodysenteriae 318:331 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 11 80 theme colonic 1921:1927 arg1 mucus 1929:1933 porcine colonic mucus 1913:1933 porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions 1913:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 5 81 theme healthy 801:807 arg1 tissues 815:821 five healthy colon tissues 796:821 five healthy colon tissues 796:821 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 9 82 theme lower 1602:1606 arg1 abundance 1608:1616 a lower abundance 1600:1616 a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1600:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 6 83 theme infected 1075:1082 arg1 pigs 1084:1087 infected pigs 1075:1087 infected pigs 1075:1087 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 2 84 from increase 303:310 arg1 mucins 350:355 mucins 350:355 mucins 350:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 2 84 from increase 303:310 arg1 sites 341:345 B. hyodysenteriae binding sites 315:345 B. hyodysenteriae binding sites on mucins 315:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 0 85 theme O-Glycans 120:128 arg1 Expression 99:108 an Increased Expression 86:108 an Increased Expression of Core-2 O-Glycans in Porcine Colon 86:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 10 86 gly glycosylation 1802:1814 arg1 pigs 1819:1822 pigs 1819:1822 pigs 1819:1822 Therefore, we conclude that B. hyodysenteriae plays a major role in regulating colonic mucin glycosylation in pigs during SD. 28301166 6 87 theme structures 1049:1058 arg1 present 1064:1070 present 1064:1070 present 1064:1070 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 87 theme structures 1049:1058 arg1 array 1033:1037 a larger array 1024:1037 a larger array of glycan structures 1024:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 88 from pigs 1084:1087 arg1 present 1064:1070 present 1064:1070 present 1064:1070 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 88 from pigs 1084:1087 arg1 array 1033:1037 a larger array 1024:1037 a larger array of glycan structures 1024:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 89 located present 1064:1070 arg2 present 1064:1070 present 1064:1070 present 1064:1070 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 89 located present 1064:1070 arg2 array 1033:1037 a larger array 1024:1037 a larger array of glycan structures 1024:1058 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 6 89 located present 1064:1070 arg1 pigs 1084:1087 infected pigs 1075:1087 infected pigs 1075:1087 In total, 94 O-glycans were identified, with healthy pigs having higher interindividual variation, although a larger array of glycan structures was present in infected pigs. 28301166 3 90 theme mucus 499:503 arg1 layer 505:509 the porcine intestinal mucus layer 476:509 the porcine intestinal mucus layer 476:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 0 91 theme Porcine 133:139 arg1 Colon 141:145 Porcine Colon 133:145 Porcine Colon 133:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 3 92 theme glycosylation 451:463 arg1 pattern 465:471 the glycosylation pattern 447:471 the glycosylation pattern of the porcine intestinal mucus layer 447:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 5 93 theme carbon 861:866 arg1 chromatography 875:888 porous graphitized carbon liquid chromatography 842:888 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 9 94 theme glycan 1427:1432 arg1 chains 1434:1439 glycan chains 1427:1439 glycan chains from infected pigs 1427:1458 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 4 95 theme colonic 566:572 arg1 O-glycome 580:588 the swine colonic mucin O-glycome 556:588 the swine colonic mucin O-glycome 556:588 We characterized the swine colonic mucin O-glycome and identified the differences in glycosylation between B. hyodysenteriae-infected and noninfected pigs. 28301166 12 96 dep potential 2061:2069 arg1 therapeutic 2071:2081 therapeutic 2071:2081 therapeutic 2071:2081 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 2 97 theme colonic 379:385 arg1 secretion 393:401 increased colonic mucin secretion 369:401 increased colonic mucin secretion 369:401 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 5 98 theme chromatography 875:888 arg1 spectrometry 902:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 9 99 contain had 1596:1598 arg2 abundance 1608:1616 a lower abundance 1600:1616 a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1600:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 9 99 contain had 1596:1598 arg1 they 1591:1594 they 1591:1594 they 1591:1594 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 3 100 theme porcine 480:486 arg1 layer 505:509 the porcine intestinal mucus layer 476:509 the porcine intestinal mucus layer 476:509 We predict that B. hyodysenteriae modifies the glycosylation pattern of the porcine intestinal mucus layer to optimize its host niche. 28301166 0 101 theme Glycosylation 53:65 arg1 Synthesis 67:75 Mucin Glycosylation Synthesis 47:75 Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon 47:145 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 5 102 theme porous 842:847 arg1 chromatography 875:888 porous graphitized carbon liquid chromatography 842:888 porous graphitized carbon liquid chromatography tandem mass spectrometry 842:913 O-Glycans were chemically released from soluble and insoluble mucins isolated from five infected and five healthy colon tissues and analyzed using porous graphitized carbon liquid chromatography tandem mass spectrometry. 28301166 12 103 from study 2047:2051 arg1 results 2029:2035 The results 2025:2035 The results from this study 2025:2051 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 9 104 contain had 1477:1479 arg1 chains 1434:1439 glycan chains 1427:1439 glycan chains from infected pigs 1427:1458 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 9 104 contain had 1477:1479 arg2 abundance 1490:1498 a higher abundance 1481:1498 a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1481:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 2 105 theme binding 333:339 arg1 sites 341:345 B. hyodysenteriae binding sites 315:345 B. hyodysenteriae binding sites on mucins 315:355 Infection with this pathogen results in an increase in B. hyodysenteriae binding sites on mucins, along with increased colonic mucin secretion. 28301166 9 106 theme higher 1483:1488 arg1 abundance 1490:1498 a higher abundance 1481:1498 a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs 1481:1706 Overall, glycan chains from infected pigs were shorter and had a higher abundance of structures that were neutral or predominantly contained NeuGc instead of NeuAc, whereas they had a lower abundance of structures that were fucosylated, acidic, or sulfated than those from noninfected pigs. 28301166 12 107 theme B. 2128:2129 arg1 interactions 2146:2157 B. hyodysenteriae interactions 2128:2157 B. hyodysenteriae interactions with the host via mucin glycans 2128:2189 The results from this study provide potential therapeutic targets and a platform for investigations of B. hyodysenteriae interactions with the host via mucin glycans. 28301166 11 108 theme host-pathogen 1997:2009 arg1 interactions 2011:2022 host-pathogen interactions 1997:2022 host-pathogen interactions 1997:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 28301166 0 109 theme hyodysenteriae 12:25 arg1 Infection 27:35 Brachyspira hyodysenteriae Infection 0:35 Brachyspira hyodysenteriae Infection 0:35 Brachyspira hyodysenteriae Infection Regulates Mucin Glycosylation Synthesis Inducing an Increased Expression of Core-2 O-Glycans in Porcine Colon. 28301166 11 110 theme porcine 1913:1919 arg1 mucus 1929:1933 porcine colonic mucus 1913:1933 porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions 1913:2022 The changes in mucin O-glycosylation thus resulted in a glycan fingerprint in porcine colonic mucus that may provide increased exposure of epitopes important for host-pathogen interactions. 26132161 6 0 theme myrAKT 1133:1138 arg1 introduction 1088:1099 The sequential introduction 1073:1099 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT 1073:1138 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 Ras 1540:1542 Ras 1540:1542 Ras 1540:1542 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 hTERT 1643:1647 hTERT 1643:1647 hTERT 1643:1647 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 N-glycans 1208:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 SV40ER 1650:1655 SV40ER 1650:1655 SV40ER 1650:1655 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 GD3 1222:1224 GD3 1222:1224 GD3 1222:1224 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 N-glycans 1632:1640 fucosylated N-glycans 1620:1640 fucosylated N-glycans 1620:1640 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 Ras 1658:1660 Ras 1658:1660 Ras 1658:1660 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 antigen-4 1451:1459 stage-specific embryonic antigen-4 1426:1459 stage-specific embryonic antigen-4 1426:1459 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 hTERT 1227:1231 hTERT 1227:1231 hTERT 1227:1231 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 expression 1363:1372 (iii) temporal expression 1348:1372 (iii) temporal expression of bisecting GlcNAc residues 1348:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 AKT 1666:1668 AKT 1666:1668 AKT 1666:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 α2,6-sialylation 1404:1419 α2,6-sialylation 1404:1419 α2,6-sialylation 1404:1419 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 1 from ganglio- 1255:1262 arg1 switching 1240:1248 (ii) switching 1235:1248 (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER) 1235:1345 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 7 2 theme tumor 1768:1772 arg1 prognosis 1774:1782 tumor prognosis 1768:1782 tumor prognosis 1768:1782 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 3 3 theme glycomic 573:580 arg1 analysis 582:589 comprehensive glycomic analysis 559:589 comprehensive glycomic analysis 559:589 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 6 4 theme globo-series 1267:1278 arg1 glycosphingolipids 1280:1297 globo-series glycosphingolipids 1267:1297 globo-series glycosphingolipids 1267:1297 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 5 theme antigen-4 1451:1459 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 6 theme Ras 1540:1542 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 7 7 theme tumor 1742:1746 arg1 diagnosis 1754:1762 tumor grade diagnosis 1742:1762 tumor grade diagnosis 1742:1762 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 6 8 theme type 1203:1206 arg1 hTERT 1227:1231 hTERT 1227:1231 hTERT 1227:1231 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 8 theme type 1203:1206 arg1 N-glycans 1208:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 4 9 theme hTERT 720:724 arg1 introduction 704:715 the serial introduction 693:715 the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT 693:754 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 10 theme SV40ER 1529:1534 arg1 suppression 1477:1487 suppression 1477:1487 suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER 1477:1534 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 11 theme AKT 1666:1668 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 12 theme surface 869:875 arg1 O-glycans 901:909 O-glycans 901:909 O-glycans 901:909 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 12 theme surface 869:875 arg1 N- 894:895 N- 894:895 N- 894:895 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 12 theme surface 869:875 arg1 glycoconjugates 877:891 cell surface glycoconjugates 864:891 cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 864:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 6 13 theme Ras 1658:1660 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 3 14 theme multistep 617:625 arg1 model 641:645 a previously described multistep tumorigenesis model 594:645 a previously described multistep tumorigenesis model 594:645 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 1 15 theme Cancer 126:131 arg1 cells 133:137 Cancer cells 126:137 Cancer cells 126:137 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 6 16 theme SV40ER 1650:1655 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 17 theme Neu5Gc 1321:1326 arg1 glycosphingolipids 1280:1297 globo-series glycosphingolipids 1267:1297 globo-series glycosphingolipids 1267:1297 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 17 theme Neu5Gc 1321:1326 arg1 appearance 1307:1316 the appearance 1303:1316 the appearance of Neu5Gc (hTERT and SV40ER) 1303:1345 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 18 from N- 894:895 arg1 glycoproteins 914:926 glycoproteins 914:926 glycoproteins 914:926 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 18 from N- 894:895 arg1 glycosphingolipids 933:950 glycosphingolipids 933:950 glycosphingolipids 933:950 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 7 19 theme transient 1693:1701 arg1 alterations 1712:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 6 20 theme hTERT 1643:1647 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 4 21 theme SV40ER 727:732 arg1 introduction 704:715 the serial introduction 693:715 the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT 693:754 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 22 theme GlcNAc 1387:1392 arg1 residues 1394:1401 bisecting GlcNAc residues 1377:1401 bisecting GlcNAc residues 1377:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 1 23 theme beneficial 352:361 arg1 approach 363:370 a beneficial approach 350:370 a beneficial approach to cancer therapy 350:388 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 6 24 theme stage-specific 1426:1439 arg1 antigen-4 1451:1459 stage-specific embryonic antigen-4 1426:1459 stage-specific embryonic antigen-4 1426:1459 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 7 25 theme sequential 1678:1687 arg1 alterations 1712:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 4 26 theme H-RasV12 735:742 arg1 introduction 704:715 the serial introduction 693:715 the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT 693:754 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 0 27 theme Specific 71:78 arg1 Patterns 94:101 Specific Glycosylation Patterns 71:101 Specific Glycosylation Patterns Related to Malignancy 71:123 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 1 28 theme different 169:177 arg1 levels 179:184 different levels 169:184 different levels 169:184 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 6 29 theme temporal 1354:1361 arg1 expression 1363:1372 (iii) temporal expression 1348:1372 (iii) temporal expression of bisecting GlcNAc residues 1348:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 30 from O-glycans 901:909 arg1 glycoproteins 914:926 glycoproteins 914:926 glycoproteins 914:926 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 30 from O-glycans 901:909 arg1 glycosphingolipids 933:950 glycosphingolipids 933:950 glycosphingolipids 933:950 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 31 theme glycosylation 996:1008 arg1 patterns 1010:1017 specific glycosylation patterns 987:1017 specific glycosylation patterns related to malignancy 987:1039 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 2 32 theme altered 425:431 arg1 glycosylation 433:445 altered glycosylation 425:445 altered glycosylation 425:445 However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. 26132161 6 33 theme glycosphingolipids 1597:1614 arg1 expression 1565:1574 expression 1565:1574 expression of (neo)lacto-series glycosphingolipids 1565:1614 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 34 theme related 1019:1025 arg1 patterns 1010:1017 specific glycosylation patterns 987:1017 specific glycosylation patterns related to malignancy 987:1039 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 6 35 theme H-RasV12 1119:1126 arg1 introduction 1088:1099 The sequential introduction 1073:1099 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT 1073:1138 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 0 36 theme Comprehensive 0:12 arg1 Glycomics 14:22 Comprehensive Glycomics 0:22 Comprehensive Glycomics of a Multistep Human Brain Tumor Model 0:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 6 37 theme SV40ER 1111:1116 arg1 introduction 1088:1099 The sequential introduction 1073:1099 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT 1073:1138 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 4 38 theme brain 781:785 arg1 grades 793:798 human brain tumor grades I-IV 775:803 human brain tumor grades I-IV 775:803 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 39 from expression 1160:1169 arg1 ganglio- 1255:1262 ganglio- 1255:1262 ganglio- 1255:1262 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 2 40 theme genetic 458:464 arg1 s 477:477 s 477:477 s 477:477 However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. 26132161 2 40 theme genetic 458:464 arg1 alteration 466:475 causal genetic alteration 451:475 causal genetic alteration(s) 451:478 However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. 26132161 5 41 theme major 847:851 arg1 classes 853:859 three major classes 841:859 three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 841:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 6 42 theme hTERT 1104:1108 arg1 introduction 1088:1099 The sequential introduction 1073:1099 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT 1073:1138 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 0 43 theme Multistep 29:37 arg1 Model 57:61 a Multistep Human Brain Tumor Model 27:61 a Multistep Human Brain Tumor Model 27:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 1 44 theme glycosylations 323:336 arg1 manipulation 301:312 manipulation 301:312 manipulation 301:312 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 1 44 theme glycosylations 323:336 arg1 elucidation 285:295 elucidation 285:295 elucidation 285:295 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 0 45 theme Model 57:61 arg1 Glycomics 14:22 Comprehensive Glycomics 0:22 Comprehensive Glycomics of a Multistep Human Brain Tumor Model 0:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 5 46 dep glycoconjugates 877:891 arg1 O-glycans 901:909 O-glycans 901:909 O-glycans 901:909 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 46 dep glycoconjugates 877:891 arg1 N- 894:895 N- 894:895 N- 894:895 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 46 dep glycoconjugates 877:891 arg1 glycoconjugates 877:891 cell surface glycoconjugates 864:891 cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 864:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 0 47 theme Brain 45:49 arg1 Model 57:61 a Multistep Human Brain Tumor Model 27:61 a Multistep Human Brain Tumor Model 27:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 7 48 theme response 1826:1833 arg1 prediction 1802:1811 the prediction 1798:1811 the prediction of treatment response 1798:1833 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 3 49 theme unique 530:535 arg1 approach 537:544 a unique approach 528:544 a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model 528:645 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 6 50 dep expression 1160:1169 arg1 i 1148:1148 i 1148:1148 i 1148:1148 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 1 51 theme normal 257:262 arg1 cells 264:268 normal cells 257:268 normal cells 257:268 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 0 52 theme Tumor 51:55 arg1 Model 57:61 a Multistep Human Brain Tumor Model 27:61 a Multistep Human Brain Tumor Model 27:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 4 53 theme human 655:659 arg1 astrocytes 661:670 Normal human astrocytes 648:670 Normal human astrocytes 648:670 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 54 theme bisecting 1377:1385 arg1 residues 1394:1401 bisecting GlcNAc residues 1377:1401 bisecting GlcNAc residues 1377:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 55 attach derived 828:834 arg1 classes 853:859 three major classes 841:859 three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 841:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 55 attach derived 828:834 arg2 glycans 820:826 More than 160 glycans 806:826 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 806:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 3 56 theme comprehensive 559:571 arg1 analysis 582:589 comprehensive glycomic analysis 559:589 comprehensive glycomic analysis 559:589 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 7 57 theme grade 1748:1752 arg1 diagnosis 1754:1762 tumor grade diagnosis 1742:1762 tumor grade diagnosis 1742:1762 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 6 58 theme GD3 1222:1224 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 59 theme glycoconjugates 877:891 arg1 classes 853:859 three major classes 841:859 three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 841:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 6 60 theme N-glycans 1208:1216 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 4 61 theme serial 697:702 arg1 introduction 704:715 the serial introduction 693:715 the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT 693:754 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 62 dep Neu5Gc 1321:1326 arg1 hTERT 1329:1333 hTERT 1329:1333 hTERT 1329:1333 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 62 dep Neu5Gc 1321:1326 arg1 SV40ER 1339:1344 SV40ER 1339:1344 SV40ER 1339:1344 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 63 theme pauci-mannose/mono-antennary 1174:1201 arg1 hTERT 1227:1231 hTERT 1227:1231 hTERT 1227:1231 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 63 theme pauci-mannose/mono-antennary 1174:1201 arg1 N-glycans 1208:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 1 64 theme different 212:220 arg1 structures 222:231 fundamentally different structures 198:231 fundamentally different structures from those expressed by normal cells 198:268 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 3 65 theme tumorigenesis 627:639 arg1 model 641:645 a previously described multistep tumorigenesis model 594:645 a previously described multistep tumorigenesis model 594:645 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 5 66 theme cell 864:867 arg1 O-glycans 901:909 O-glycans 901:909 O-glycans 901:909 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 66 theme cell 864:867 arg1 N- 894:895 N- 894:895 N- 894:895 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 5 66 theme cell 864:867 arg1 glycoconjugates 877:891 cell surface glycoconjugates 864:891 cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) 864:951 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 6 67 theme switching 1240:1248 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 68 theme expression 1363:1372 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 3 69 theme described 607:615 arg1 model 641:645 a previously described multistep tumorigenesis model 594:645 a previously described multistep tumorigenesis model 594:645 Here, we employed a unique approach that applies comprehensive glycomic analysis to a previously described multistep tumorigenesis model. 26132161 1 70 from those 238:242 arg1 structures 222:231 fundamentally different structures 198:231 fundamentally different structures from those expressed by normal cells 198:268 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 6 71 theme increased 1555:1563 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 72 theme sequential 1077:1086 arg1 introduction 1088:1099 The sequential introduction 1073:1099 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT 1073:1138 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 73 gly glycoproteins 914:926 arg1 glycoproteins 914:926 glycoproteins 914:926 glycoproteins 914:926 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 4 74 theme myrAKT 749:754 arg1 introduction 704:715 the serial introduction 693:715 the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT 693:754 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 75 theme residues 1394:1401 arg1 Ras 1540:1542 Ras 1540:1542 Ras 1540:1542 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 hTERT 1643:1647 hTERT 1643:1647 hTERT 1643:1647 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 N-glycans 1208:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 pauci-mannose/mono-antennary type N-glycans 1174:1216 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 SV40ER 1650:1655 SV40ER 1650:1655 SV40ER 1650:1655 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 GD3 1222:1224 GD3 1222:1224 GD3 1222:1224 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 N-glycans 1632:1640 fucosylated N-glycans 1620:1640 fucosylated N-glycans 1620:1640 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 Ras 1658:1660 Ras 1658:1660 Ras 1658:1660 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 antigen-4 1451:1459 stage-specific embryonic antigen-4 1426:1459 stage-specific embryonic antigen-4 1426:1459 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 hTERT 1227:1231 hTERT 1227:1231 hTERT 1227:1231 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 expression 1363:1372 (iii) temporal expression 1348:1372 (iii) temporal expression of bisecting GlcNAc residues 1348:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 AKT 1666:1668 AKT 1666:1668 AKT 1666:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 α2,6-sialylation 1404:1419 α2,6-sialylation 1404:1419 α2,6-sialylation 1404:1419 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 75 theme residues 1394:1401 arg1 switching 1240:1248 (ii) switching 1235:1248 (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER) 1235:1345 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 7 76 theme glycomic 1703:1710 arg1 alterations 1712:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations 1672:1722 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 0 77 theme Glycosylation 80:92 arg1 Patterns 94:101 Specific Glycosylation Patterns 71:101 Specific Glycosylation Patterns Related to Malignancy 71:123 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 4 78 dep grades 793:798 arg1 I-IV 800:803 I-IV 800:803 human brain tumor grades I-IV 775:803 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 79 dep increased 1555:1563 arg1 iv 1551:1552 iv 1551:1552 iv 1551:1552 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 80 dep switching 1240:1248 arg1 ii 1236:1237 ii 1236:1237 ii 1236:1237 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 0 81 theme Related 103:109 arg1 Patterns 94:101 Specific Glycosylation Patterns 71:101 Specific Glycosylation Patterns Related to Malignancy 71:123 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 1 82 theme cancer 375:380 arg1 therapy 382:388 cancer therapy 375:388 cancer therapy 375:388 Cancer cells frequently express glycans at different levels and/or with fundamentally different structures from those expressed by normal cells, and therefore elucidation and manipulation of these glycosylations may provide a beneficial approach to cancer therapy. 26132161 6 83 dep expression 1363:1372 arg1 iii 1349:1351 iii 1349:1351 iii 1349:1351 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 84 theme fucosylated 1620:1630 arg1 N-glycans 1632:1640 fucosylated N-glycans 1620:1640 fucosylated N-glycans 1620:1640 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 5 85 theme specific 987:994 arg1 patterns 1010:1017 specific glycosylation patterns 987:1017 specific glycosylation patterns related to malignancy 987:1039 More than 160 glycans derived from three major classes of cell surface glycoconjugates (N- and O-glycans on glycoproteins, and glycosphingolipids) were quantitatively explored, and specific glycosylation patterns related to malignancy were systematically identified. 26132161 4 86 theme tumor 787:791 arg1 grades 793:798 human brain tumor grades I-IV 775:803 human brain tumor grades I-IV 775:803 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 87 theme N-glycans 1632:1640 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 88 theme lacto-series 1584:1595 arg1 neo 1580:1582 neo 1580:1582 neo 1580:1582 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 88 theme lacto-series 1584:1595 arg1 glycosphingolipids 1597:1614 (neo)lacto-series glycosphingolipids 1579:1614 (neo)lacto-series glycosphingolipids 1579:1614 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 89 theme embryonic 1441:1449 arg1 antigen-4 1451:1459 stage-specific embryonic antigen-4 1426:1459 stage-specific embryonic antigen-4 1426:1459 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 4 90 theme human 775:779 arg1 grades 793:798 human brain tumor grades I-IV 775:803 human brain tumor grades I-IV 775:803 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 2 91 theme causal 451:456 arg1 s 477:477 s 477:477 s 477:477 However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. 26132161 2 91 theme causal 451:456 arg1 alteration 466:475 causal genetic alteration 451:475 causal genetic alteration(s) 451:478 However, the relationship between altered glycosylation and causal genetic alteration(s) is only partially understood. 26132161 0 92 theme Human 39:43 arg1 Model 57:61 a Multistep Human Brain Tumor Model 27:61 a Multistep Human Brain Tumor Model 27:61 Comprehensive Glycomics of a Multistep Human Brain Tumor Model Reveals Specific Glycosylation Patterns Related to Malignancy. 26132161 6 93 theme α2,6-sialylation 1404:1419 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 94 theme temporal 1151:1158 arg1 expression 1160:1169 (i) temporal expression 1147:1169 (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT 1147:1668 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 6 95 gly fucosylated 1620:1630 arg1 N-glycans 1632:1640 fucosylated N-glycans 1620:1640 fucosylated N-glycans 1620:1640 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 26132161 7 96 theme treatment 1816:1824 arg1 response 1826:1833 treatment response 1816:1833 treatment response 1816:1833 These sequential and transient glycomic alterations may be useful for tumor grade diagnosis and tumor prognosis, and also for the prediction of treatment response. 26132161 4 97 theme Normal 648:653 arg1 astrocytes 661:670 Normal human astrocytes 648:670 Normal human astrocytes 648:670 Normal human astrocytes were transformed via the serial introduction of hTERT, SV40ER, H-RasV12, and myrAKT, thereby mimicking human brain tumor grades I-IV. 26132161 6 98 gly α2,6-sialylation 1404:1419 arg1 residues 1394:1401 bisecting GlcNAc residues 1377:1401 bisecting GlcNAc residues 1377:1401 The sequential introduction of hTERT, SV40ER, H-RasV12, and myrAKT led to (i) temporal expression of pauci-mannose/mono-antennary type N-glycans and GD3 (hTERT); (ii) switching from ganglio- to globo-series glycosphingolipids and the appearance of Neu5Gc (hTERT and SV40ER); (iii) temporal expression of bisecting GlcNAc residues, α2,6-sialylation, and stage-specific embryonic antigen-4, accompanied by suppression of core 2 O-glycan biosynthesis (hTERT, SV40ER and Ras); and (iv) increased expression of (neo)lacto-series glycosphingolipids and fucosylated N-glycans (hTERT, SV40ER, Ras and AKT). 27267257 4 0 theme linker 695:700 arg1 stability 702:710 the linker stability 691:710 the relationships of glycosylating agents and nucleophiles as well as the linker stability 621:710 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 4 1 theme target 561:566 arg1 structures 568:577 the target structures 557:577 the target structures 557:577 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 4 2 theme essential 580:588 arg1 insights 590:597 essential insights 580:597 essential insights 580:597 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 3 3 theme α-fucoses 373:381 arg1 incorporation 356:368 The incorporation 352:368 The incorporation of α-fucoses 352:381 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 4 4 dep preparing 547:555 arg1 addition 535:542 addition 535:542 addition 535:542 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 3 5 dep I 410:410 arg1 required 491:498 required 491:498 required incorporation of a 3-O-sulfate 491:529 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 3 5 dep I 410:410 arg1 prepared 451:458 prepared 451:458 were prepared 446:458 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 3 6 theme 3-O-sulfate 519:529 arg1 incorporation 500:512 incorporation 500:512 incorporation of a 3-O-sulfate 500:529 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 4 7 theme agents 656:661 arg1 relationships 625:637 the relationships 621:637 the relationships of glycosylating agents and nucleophiles as well as the linker stability 621:710 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 4 7 theme agents 656:661 arg1 stability 702:710 the linker stability 691:710 the relationships of glycosylating agents and nucleophiles as well as the linker stability 621:710 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 0 8 theme Glycan 10:15 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Complex 0:35 Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants. 27267257 0 9 theme Automated 0:8 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Complex 0:35 Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants. 27267257 4 10 gly glycosylating 642:654 arg0 agents 656:661 glycosylating agents 642:661 glycosylating agents 642:661 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 4 11 theme nucleophiles 667:678 arg1 relationships 625:637 the relationships 621:637 the relationships of glycosylating agents and nucleophiles as well as the linker stability 621:710 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 4 11 theme nucleophiles 667:678 arg1 stability 702:710 the linker stability 691:710 the relationships of glycosylating agents and nucleophiles as well as the linker stability 621:710 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 3 12 theme pentasaccharide 469:483 arg1 HNK-1 485:489 the pentasaccharide HNK-1 465:489 the pentasaccharide HNK-1 465:489 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 4 13 theme glycosylating 642:654 arg1 agents 656:661 glycosylating agents 642:661 glycosylating agents 642:661 In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability. 27267257 1 14 theme many 175:178 arg1 glycans 180:186 many glycans 175:186 many glycans 175:186 Lactotetraosyl (Lc4) and neo-lactotetraosyl (nLc4) are backbones that are common to many glycans. 27267257 3 15 theme -galactose 426:435 arg1 epitopes 437:444 α(1,3)-galactose epitopes 420:444 α(1,3)-galactose epitopes 420:444 The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. 27267257 0 16 theme Complex 29:35 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Complex 0:35 Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants. 27267257 2 17 theme automated 195:203 arg1 assembly 212:219 automated glycan assembly 195:219 automated glycan assembly 195:219 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 27267257 2 18 theme glycan 205:210 arg1 assembly 212:219 automated glycan assembly 195:219 automated glycan assembly 195:219 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 27267257 2 19 theme relevance 341:349 arg1 glycans 319:325 glycans 319:325 glycans of biological relevance 319:349 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 27267257 2 20 theme biological 330:339 arg1 relevance 341:349 biological relevance 330:349 biological relevance 330:349 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 27267257 0 21 theme Group 71:75 arg1 Determinants 77:88 Blood Group Determinants 65:88 Blood Group Determinants 65:88 Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants. 27267257 2 22 theme core 235:238 arg1 structures 240:249 these common core structures 222:249 these common core structures 222:249 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 27267257 0 23 theme Blood 65:69 arg1 Determinants 77:88 Blood Group Determinants 65:88 Blood Group Determinants 65:88 Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants. 27267257 2 24 theme common 228:233 arg1 structures 240:249 these common core structures 222:249 these common core structures 222:249 Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. 28619057 4 0 theme first 597:601 arg1 investigation 612:624 the first detailed investigation 593:624 the first detailed investigation of the cell wall composition of collenchyma from any plant 593:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 4 0 theme first 597:601 arg1 This 585:588 This 585:588 This 585:588 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 7 1 theme HEPES 1097:1101 arg1 fraction 1118:1125 a HEPES buffer soluble fraction 1095:1125 a HEPES buffer soluble fraction 1095:1125 Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. 28619057 13 2 from residue 2157:2163 arg1 predominant 2132:2142 predominant 2132:2142 predominant 2132:2142 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 3 3 theme peripheral 514:523 arg1 collenchyma 525:535 peripheral collenchyma 514:535 peripheral collenchyma 514:535 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 15 4 theme rigid 2451:2455 arg1 polysaccharides 2472:2486 the rigid and semi-rigid polysaccharides 2447:2486 the rigid and semi-rigid polysaccharides 2447:2486 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 13 5 theme 4 M 2055:2057 arg1 rich 2076:2079 rich 2076:2079 rich 2076:2079 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 5 theme 4 M 2055:2057 arg1 fraction 2063:2070 the 4 M KOH fraction 2051:2070 the 4 M KOH fraction 2051:2070 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 20 6 contain contained 3179:3187 arg1 fraction 3170:3177 the HEPES-soluble fraction 3152:3177 the HEPES-soluble fraction 3152:3177 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 20 6 contain contained 3179:3187 arg2 heteroxylans 3207:3218 heteroxylans 3207:3218 heteroxylans 3207:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 20 6 contain contained 3179:3187 arg2 amount 3197:3202 a large amount 3189:3202 a large amount of heteroxylans 3189:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 5 7 theme Celery 686:691 arg1 petioles 693:700 Celery petioles 686:700 Celery petioles 686:700 Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. 28619057 11 8 theme esterification 1771:1784 arg1 degree 1754:1759 a higher degree 1745:1759 a higher degree of methyl esterification 1745:1784 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 13 9 contain had 2024:2026 arg1 fraction 2015:2022 the 1 M KOH fraction 2003:2022 the 1 M KOH fraction 2003:2022 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 9 contain had 2024:2026 arg2 heteroxylans 2037:2048 abundant heteroxylans 2028:2048 abundant heteroxylans 2028:2048 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 10 10 from segments 1555:1562 arg1 CWs 1536:1538 CWs 1536:1538 CWs from different segments 1536:1562 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 16 11 theme cell 2727:2730 arg1 walls 2732:2736 the cell walls 2723:2736 the cell walls 2723:2736 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 4 12 from plant 679:683 arg1 composition 643:653 the cell wall composition 629:653 the cell wall composition of collenchyma from any plant 629:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 4 12 from plant 679:683 arg1 investigation 612:624 the first detailed investigation 593:624 the first detailed investigation of the cell wall composition of collenchyma from any plant 593:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 4 12 from plant 679:683 arg1 This 585:588 This 585:588 This 585:588 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 1 13 theme herbaceous 168:177 arg1 plants 179:184 many herbaceous plants 163:184 many herbaceous plants 163:184 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 18 14 theme celery 2942:2947 arg1 CWs 2960:2962 celery parenchyma CWs 2942:2962 celery parenchyma CWs 2942:2962 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 14 15 theme linkage 2254:2260 arg1 analysis 2262:2269 the linkage analysis 2250:2269 the linkage analysis 2250:2269 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 2 16 theme polysaccharide 303:316 arg1 compositions 318:329 similar polysaccharide compositions 295:329 similar polysaccharide compositions 295:329 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 15 17 theme segments 2512:2519 arg1 CWs 2495:2497 the CWs 2491:2497 the CWs of the three segments 2491:2519 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 12 18 theme large 1916:1920 arg1 heteroxylans 1933:1944 heteroxylans 1933:1944 heteroxylans 1933:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 12 18 theme large 1916:1920 arg1 amounts 1922:1928 large amounts 1916:1928 large amounts of heteroxylans 1916:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 6 19 theme lower 965:969 arg1 segments 937:944 three equal segments 925:944 three equal segments 925:944 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 17 20 theme similar 2803:2809 arg1 compositions 2826:2837 similar polysaccharide compositions 2803:2837 similar polysaccharide compositions 2803:2837 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 6 21 theme middle 954:959 arg1 segments 937:944 three equal segments 925:944 three equal segments 925:944 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 11 22 from pectin 1702:1707 arg1 CWs 1716:1718 the CWs 1712:1718 the CWs of the upper segment 1712:1739 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 14 23 from CWs 2338:2340 arg1 present 2297:2303 present 2297:2303 present 2297:2303 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 17 24 theme CONCLUSIONS 2763:2773 arg1 CWs 2794:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 10 25 theme similar 1568:1574 arg1 compositions 1576:1587 similar compositions 1568:1587 similar compositions 1568:1587 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 5 26 theme upper 834:838 arg1 region 840:845 the upper region 830:845 the upper region 830:845 Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. 28619057 12 27 theme HEPES-soluble 1869:1881 arg1 fractions 1883:1891 the HEPES-soluble fractions 1865:1891 the HEPES-soluble fractions 1865:1891 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 6 28 theme upper 947:951 arg1 segments 937:944 three equal segments 925:944 three equal segments 925:944 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 11 29 theme upper 1727:1731 arg1 segment 1733:1739 the upper segment 1723:1739 the upper segment 1723:1739 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 9 30 theme Monosaccharide 1204:1217 arg1 compositions 1219:1230 Monosaccharide compositions 1204:1230 Monosaccharide compositions of the CWs 1204:1241 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 2 31 theme solid-state 210:220 arg1 NMR 222:224 solid-state NMR 210:224 solid-state NMR 210:224 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 19 32 theme arabinan 3037:3044 arg1 chains 3051:3056 the arabinan side chains 3033:3056 the arabinan side chains of RG-I 3033:3064 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 33 from variation 3076:3084 arg1 segments 3128:3135 the different segments 3114:3135 the different segments 3114:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 33 from variation 3076:3084 arg1 CWs 3105:3107 the collenchyma CWs 3089:3107 the collenchyma CWs from the different segments 3089:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 13 34 theme KOH 2011:2013 arg1 fraction 2015:2022 the 1 M KOH fraction 2003:2022 the 1 M KOH fraction 2003:2022 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 2 35 theme cell 269:272 arg1 CWs 281:283 CWs 281:283 CWs 281:283 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 2 35 theme cell 269:272 arg1 walls 274:278 collenchyma cell walls 257:278 collenchyma cell walls (CWs) 257:284 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 15 36 theme magic 2367:2371 arg1 angle 2373:2377 magic angle 2367:2377 magic angle spinning (CP/MAS) NMR spectroscopy 2367:2412 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 9 37 theme other 1437:1441 arg1 polysaccharides 1443:1457 other polysaccharides 1437:1457 other polysaccharides 1437:1457 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 38 theme CWs 1239:1241 arg1 compositions 1219:1230 Monosaccharide compositions 1204:1230 Monosaccharide compositions of the CWs 1204:1241 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 1 39 theme support 144:150 arg1 Collenchyma 109:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 1 39 theme support 144:150 arg1 tissue 152:157 a mechanical support tissue 131:157 a mechanical support tissue for many herbaceous plants 131:184 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 3 40 from petioles 549:556 arg1 compositions 454:465 compositions 454:465 compositions 454:465 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 3 40 from petioles 549:556 arg1 structures 471:480 structures 471:480 structures 471:480 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 5 41 theme matured 782:788 arg1 elongation 790:799 matured elongation 782:799 matured elongation 782:799 Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. 28619057 5 41 theme matured 782:788 arg1 they 777:780 they 777:780 they 777:780 Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. 28619057 6 42 theme collenchyma 987:997 arg1 strands 999:1005 peripheral collenchyma strands 976:1005 peripheral collenchyma strands 976:1005 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 16 43 theme detailed 2682:2689 arg1 structure 2691:2699 the detailed structure 2678:2699 the detailed structure 2678:2699 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 1 44 theme BACKGROUND 98:107 arg1 Collenchyma 109:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 1 44 theme BACKGROUND 98:107 arg1 tissue 152:157 a mechanical support tissue 131:157 a mechanical support tissue for many herbaceous plants 131:184 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 5 45 theme early 756:760 arg1 growth 762:767 early growth 756:767 early growth 756:767 Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. 28619057 12 46 theme anticipated 1830:1840 arg1 pectins 1856:1862 the anticipated water-soluble pectins 1826:1862 the anticipated water-soluble pectins 1826:1862 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 15 47 theme apparent 2424:2431 arg1 difference 2433:2442 no apparent difference 2421:2442 no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments 2421:2519 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 2 48 located found 349:353 arg2 those 334:338 those 334:338 those 334:338 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 2 48 located found 349:353 arg1 walls 383:387 eudicotyledon parenchyma walls 358:387 eudicotyledon parenchyma walls 358:387 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 13 49 from predominant 2132:2142 arg1 residue 2157:2163 the final residue 2147:2163 the final residue 2147:2163 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 17 50 theme parenchyma 2861:2870 arg1 CWs 2872:2874 most eudicotyledon parenchyma CWs 2842:2874 most eudicotyledon parenchyma CWs 2842:2874 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 2 51 theme detailed 397:404 arg1 analysis 415:422 no detailed chemical analysis 394:422 no detailed chemical analysis 394:422 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 17 52 theme Celery 2775:2780 arg1 CWs 2794:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 19 53 theme different 3118:3126 arg1 segments 3128:3135 the different segments 3114:3135 the different segments 3114:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 0 54 theme Polysaccharide 0:13 arg1 compositions 15:26 Polysaccharide compositions 0:26 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles 0:95 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 9 55 theme polysaccharide 1284:1297 arg1 homogalacturonan 1305:1320 the most abundant polysaccharide [with homogalacturonan 1266:1320 the most abundant polysaccharide [with homogalacturonan (HG) 1266:1325 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 55 theme polysaccharide 1284:1297 arg1 pectin 1255:1260 pectin 1255:1260 pectin 1255:1260 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 55 theme polysaccharide 1284:1297 arg1 HG 1323:1324 HG 1323:1324 HG 1323:1324 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 15 56 theme NMR 2397:2399 arg1 CP/MAS 2389:2394 CP/MAS 2389:2394 CP/MAS 2389:2394 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 15 56 theme NMR 2397:2399 arg1 spectroscopy 2401:2412 (CP/MAS) NMR spectroscopy 2388:2412 (CP/MAS) NMR spectroscopy 2388:2412 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 9 57 theme heteroxylans 1504:1515 arg1 heteroxylans 1504:1515 heteroxylans 1504:1515 heteroxylans 1504:1515 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 57 theme heteroxylans 1504:1515 arg1 amounts 1493:1499 smaller amounts 1485:1499 smaller amounts of heteroxylans and heteromannans 1485:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 57 theme heteroxylans 1504:1515 arg1 heteromannans 1521:1533 heteromannans 1521:1533 heteromannans 1521:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 16 58 theme mobile 2621:2626 arg1 polysaccharides 2628:2642 highly mobile polysaccharides 2614:2642 highly mobile polysaccharides 2614:2642 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 16 59 with excitation 2535:2544 arg1 spectroscopy 2586:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 17 60 theme eudicotyledon 2847:2859 arg1 CWs 2872:2874 most eudicotyledon parenchyma CWs 2842:2874 most eudicotyledon parenchyma CWs 2842:2874 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 9 61 theme heteromannans 1521:1533 arg1 heteroxylans 1504:1515 heteroxylans 1504:1515 heteroxylans 1504:1515 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 61 theme heteromannans 1521:1533 arg1 amounts 1493:1499 smaller amounts 1485:1499 smaller amounts of heteroxylans and heteromannans 1485:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 61 theme heteromannans 1521:1533 arg1 heteromannans 1521:1533 heteromannans 1521:1533 heteromannans 1521:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 10 62 theme upper 1609:1613 arg1 segments 1615:1622 the upper segments 1605:1622 the upper segments 1605:1622 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 0 63 theme cell 43:46 arg1 walls 48:52 collenchyma cell walls 31:52 collenchyma cell walls 31:52 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 6 64 theme Mature 873:878 arg1 petioles 898:905 Mature, fully elongated, petioles 873:905 Mature, fully elongated, petioles 873:905 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 13 65 theme CDTA 1951:1954 arg1 rich 1982:1985 rich 1982:1985 rich 1982:1985 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 65 theme CDTA 1951:1954 arg1 fractions 1967:1975 The CDTA and Na2CO3 fractions 1947:1975 fractions 1967:1975 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 3 66 theme wall 490:493 arg1 polysaccharides 495:509 cell wall polysaccharides 485:509 cell wall polysaccharides of peripheral collenchyma 485:535 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 7 67 theme soluble 1110:1116 arg1 fraction 1118:1125 a HEPES buffer soluble fraction 1095:1125 a HEPES buffer soluble fraction 1095:1125 Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. 28619057 16 68 theme spinning 2563:2570 arg1 spectroscopy 2586:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 13 69 attach predominant 2132:2142 arg1 residue 2157:2163 the final residue 2147:2163 the final residue 2147:2163 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 69 attach predominant 2132:2142 arg2 cellulose 2118:2126 cellulose 2118:2126 cellulose 2118:2126 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 10 70 theme more 1637:1640 arg1 pectin 1642:1647 slightly more pectin 1628:1647 slightly more pectin 1628:1647 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 18 71 theme collenchyma 2893:2903 arg1 CWs 2905:2907 celery collenchyma CWs 2886:2907 celery collenchyma CWs 2886:2907 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 10 72 contain had 1564:1566 arg2 compositions 1576:1587 similar compositions 1568:1587 similar compositions 1568:1587 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 10 72 contain had 1564:1566 arg1 CWs 1536:1538 CWs 1536:1538 CWs from different segments 1536:1562 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 13 73 theme final 2151:2155 arg1 residue 2157:2163 the final residue 2147:2163 the final residue 2147:2163 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 2 74 theme Previous 187:194 arg1 work 196:199 Previous work 187:199 Previous work based on solid-state NMR and immunomicroscopy 187:245 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 14 75 theme most 2308:2311 arg1 CWs 2338:2340 most eudicotyledon parenchyma CWs 2308:2340 most eudicotyledon parenchyma CWs 2308:2340 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 10 76 theme lower 1669:1673 arg1 segments 1679:1686 the lower two segments 1665:1686 the lower two segments 1665:1686 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 20 77 theme heteroxylans 3207:3218 arg1 heteroxylans 3207:3218 heteroxylans 3207:3218 heteroxylans 3207:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 20 77 theme heteroxylans 3207:3218 arg1 amount 3197:3202 a large amount 3189:3202 a large amount of heteroxylans 3189:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 7 78 theme buffer 1103:1108 arg1 fraction 1118:1125 a HEPES buffer soluble fraction 1095:1125 a HEPES buffer soluble fraction 1095:1125 Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. 28619057 6 79 theme equal 931:935 arg1 segments 937:944 three equal segments 925:944 three equal segments 925:944 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 20 80 theme large 3191:3195 arg1 heteroxylans 3207:3218 heteroxylans 3207:3218 heteroxylans 3207:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 20 80 theme large 3191:3195 arg1 amount 3197:3202 a large amount 3189:3202 a large amount of heteroxylans 3189:3218 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 13 81 from rich 1982:1985 arg1 RG-I 1997:2000 RG-I 1997:2000 RG-I 1997:2000 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 81 from rich 1982:1985 arg1 HG 1990:1991 HG 1990:1991 HG 1990:1991 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 4 82 theme cell 633:636 arg1 composition 643:653 the cell wall composition 629:653 the cell wall composition of collenchyma from any plant 629:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 19 83 theme pectin 3004:3009 arg1 esterification 2986:2999 methyl esterification 2979:2999 methyl esterification of pectin 2979:3009 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 2 84 theme eudicotyledon 358:370 arg1 walls 383:387 eudicotyledon parenchyma walls 358:387 eudicotyledon parenchyma walls 358:387 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 18 85 theme higher 2919:2924 arg1 content 2929:2935 much higher XG content 2914:2935 much higher XG content 2914:2935 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 20 86 theme HEPES-soluble 3156:3168 arg1 fraction 3170:3177 the HEPES-soluble fraction 3152:3177 the HEPES-soluble fraction 3152:3177 Unexpectedly, the HEPES-soluble fraction contained a large amount of heteroxylans. 28619057 9 87 with abundant 1332:1339 arg1 heteroxylans 1504:1515 heteroxylans 1504:1515 heteroxylans 1504:1515 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 87 with abundant 1332:1339 arg1 amounts 1493:1499 smaller amounts 1485:1499 smaller amounts of heteroxylans and heteromannans 1485:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 87 with abundant 1332:1339 arg1 heteromannans 1521:1533 heteromannans 1521:1533 heteromannans 1521:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 13 88 theme KOH 2059:2061 arg1 rich 2076:2079 rich 2076:2079 rich 2076:2079 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 88 theme KOH 2059:2061 arg1 fraction 2063:2070 the 4 M KOH fraction 2051:2070 the 4 M KOH fraction 2051:2070 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 19 89 theme esterification 2986:2999 arg1 degree 2969:2974 The degree 2965:2974 The degree of methyl esterification of pectin 2965:3009 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 89 theme esterification 2986:2999 arg1 structures 3019:3028 the structures 3015:3028 the structures of the arabinan side chains of RG-I 3015:3064 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 4 90 theme detailed 603:610 arg1 investigation 612:624 the first detailed investigation 593:624 the first detailed investigation of the cell wall composition of collenchyma from any plant 593:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 4 90 theme detailed 603:610 arg1 This 585:588 This 585:588 This 585:588 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 17 91 contain have 2798:2801 arg1 CWs 2794:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 17 91 contain have 2798:2801 arg2 compositions 2826:2837 similar polysaccharide compositions 2803:2837 similar polysaccharide compositions 2803:2837 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 3 92 theme celery 542:547 arg1 petioles 549:556 celery petioles 542:556 celery petioles 542:556 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 1 93 theme many 163:166 arg1 plants 179:184 many herbaceous plants 163:184 many herbaceous plants 163:184 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 12 94 theme heteroxylans 1933:1944 arg1 heteroxylans 1933:1944 heteroxylans 1933:1944 heteroxylans 1933:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 12 94 theme heteroxylans 1933:1944 arg1 amounts 1922:1928 large amounts 1916:1928 large amounts of heteroxylans 1916:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 2 95 dep suggested 247:255 arg1 have 290:293 have 290:293 suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls 247:387 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 10 96 theme different 1545:1553 arg1 segments 1555:1562 different segments 1545:1562 different segments 1545:1562 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 19 97 theme RG-I 3061:3064 arg1 chains 3051:3056 the arabinan side chains 3033:3056 the arabinan side chains of RG-I 3033:3064 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 10 98 contain had 1624:1626 arg1 those 1594:1598 those 1594:1598 those 1594:1598 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 10 98 contain had 1624:1626 arg2 pectin 1642:1647 slightly more pectin 1628:1647 slightly more pectin 1628:1647 CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. 28619057 19 99 theme side 3046:3049 arg1 chains 3051:3056 the arabinan side chains 3033:3056 the arabinan side chains of RG-I 3033:3064 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 100 theme chains 3051:3056 arg1 degree 2969:2974 The degree 2965:2974 The degree of methyl esterification of pectin 2965:3009 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 100 theme chains 3051:3056 arg1 structures 3019:3028 the structures 3015:3028 the structures of the arabinan side chains of RG-I 3015:3064 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 6 101 attach isolated 1007:1014 arg2 segments 937:944 three equal segments 925:944 three equal segments 925:944 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 6 101 attach isolated 1007:1014 arg1 each 1021:1024 each 1021:1024 each 1021:1024 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 17 102 theme collenchyma 2782:2792 arg1 CWs 2794:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs 2763:2796 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 4 103 theme composition 643:653 arg1 investigation 612:624 the first detailed investigation 593:624 the first detailed investigation of the cell wall composition of collenchyma from any plant 593:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 4 103 theme composition 643:653 arg1 This 585:588 This 585:588 This 585:588 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 13 104 from RG-I 1997:2000 arg1 rich 1982:1985 rich 1982:1985 rich 1982:1985 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 104 from RG-I 1997:2000 arg1 fractions 1967:1975 The CDTA and Na2CO3 fractions 1947:1975 fractions 1967:1975 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 11 105 theme methyl 1764:1769 arg1 esterification 1771:1784 methyl esterification 1764:1784 methyl esterification 1764:1784 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 13 106 theme abundant 2028:2035 arg1 heteroxylans 2037:2048 abundant heteroxylans 2028:2048 abundant heteroxylans 2028:2048 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 14 107 theme parenchyma 2327:2336 arg1 CWs 2338:2340 most eudicotyledon parenchyma CWs 2308:2340 most eudicotyledon parenchyma CWs 2308:2340 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 4 108 theme collenchyma 658:668 arg1 composition 643:653 the cell wall composition 629:653 the cell wall composition of collenchyma from any plant 629:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 13 109 from HG 1990:1991 arg1 rich 1982:1985 rich 1982:1985 rich 1982:1985 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 109 from HG 1990:1991 arg1 fractions 1967:1975 The CDTA and Na2CO3 fractions 1947:1975 fractions 1967:1975 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 16 110 theme magic-angle 2551:2561 arg1 spectroscopy 2586:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 11 111 theme segment 1733:1739 arg1 CWs 1716:1718 the CWs 1712:1718 the CWs of the upper segment 1712:1739 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 7 112 theme Cell 1027:1030 arg1 CWs 1039:1041 CWs 1039:1041 CWs 1039:1041 Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. 28619057 7 112 theme Cell 1027:1030 arg1 walls 1032:1036 Cell walls 1027:1036 Cell walls (CWs) 1027:1042 Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. 28619057 13 113 theme 1 M 2007:2009 arg1 fraction 2015:2022 the 1 M KOH fraction 2003:2022 the 1 M KOH fraction 2003:2022 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 12 114 contain contained 1906:1914 arg2 amounts 1922:1928 large amounts 1916:1928 large amounts of heteroxylans 1916:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 12 114 contain contained 1906:1914 arg1 fractions 1883:1891 the HEPES-soluble fractions 1865:1891 the HEPES-soluble fractions 1865:1891 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 12 114 contain contained 1906:1914 arg2 heteroxylans 1933:1944 heteroxylans 1933:1944 heteroxylans 1933:1944 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 2 115 theme similar 295:301 arg1 compositions 318:329 similar polysaccharide compositions 295:329 similar polysaccharide compositions 295:329 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 15 116 with polarization 2349:2360 arg1 angle 2373:2377 magic angle 2367:2377 magic angle spinning (CP/MAS) NMR spectroscopy 2367:2412 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 13 117 theme Na2CO3 1960:1965 arg1 rich 1982:1985 rich 1982:1985 rich 1982:1985 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 117 theme Na2CO3 1960:1965 arg1 fractions 1967:1975 The CDTA and Na2CO3 fractions 1947:1975 fractions 1967:1975 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 0 118 from petioles 88:95 arg1 compositions 15:26 Polysaccharide compositions 0:26 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles 0:95 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 14 119 from present 2297:2303 arg1 CWs 2338:2340 most eudicotyledon parenchyma CWs 2308:2340 most eudicotyledon parenchyma CWs 2308:2340 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 11 120 theme higher 1747:1752 arg1 degree 1754:1759 a higher degree 1745:1759 a higher degree of methyl esterification 1745:1784 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 14 121 theme xyloglucans 2188:2198 arg1 heteroxylans 2201:2212 heteroxylans 2201:2212 heteroxylans 2201:2212 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 14 121 theme xyloglucans 2188:2198 arg1 structures 2170:2179 The structures 2166:2179 The structures of the xyloglucans 2166:2198 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 14 121 theme xyloglucans 2188:2198 arg1 heteromannans 2218:2230 heteromannans 2218:2230 heteromannans 2218:2230 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 14 121 theme xyloglucans 2188:2198 arg1 similar 2280:2286 similar 2280:2286 similar 2280:2286 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 2 122 theme collenchyma 257:267 arg1 CWs 281:283 CWs 281:283 CWs 281:283 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 2 122 theme collenchyma 257:267 arg1 walls 274:278 collenchyma cell walls 257:278 collenchyma cell walls (CWs) 257:284 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 1 123 theme mechanical 133:142 arg1 Collenchyma 109:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma 98:119 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 1 123 theme mechanical 133:142 arg1 tissue 152:157 a mechanical support tissue 131:157 a mechanical support tissue for many herbaceous plants 131:184 BACKGROUND Collenchyma serves as a mechanical support tissue for many herbaceous plants. 28619057 15 124 from difference 2433:2442 arg1 polysaccharides 2472:2486 the rigid and semi-rigid polysaccharides 2447:2486 the rigid and semi-rigid polysaccharides 2447:2486 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 15 124 from difference 2433:2442 arg1 CWs 2495:2497 the CWs 2491:2497 the CWs of the three segments 2491:2519 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 15 125 theme Cross 2343:2347 arg1 polarization 2349:2360 Cross polarization 2343:2360 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy 2343:2412 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 16 126 theme NMR 2582:2584 arg1 spectroscopy 2586:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 13 127 from heteromannans 2099:2111 arg1 rich 2076:2079 rich 2076:2079 rich 2076:2079 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 127 from heteromannans 2099:2111 arg1 fraction 2063:2070 the 4 M KOH fraction 2051:2070 the 4 M KOH fraction 2051:2070 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 4 128 from investigation 612:624 arg1 plant 679:683 any plant 675:683 any plant 675:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 18 129 contain have 2909:2912 arg2 content 2929:2935 much higher XG content 2914:2935 much higher XG content 2914:2935 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 18 129 contain have 2909:2912 arg1 CWs 2905:2907 celery collenchyma CWs 2886:2907 celery collenchyma CWs 2886:2907 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 12 130 theme water-soluble 1842:1854 arg1 pectins 1856:1862 the anticipated water-soluble pectins 1826:1862 the anticipated water-soluble pectins 1826:1862 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 6 131 theme peripheral 976:985 arg1 strands 999:1005 peripheral collenchyma strands 976:1005 peripheral collenchyma strands 976:1005 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 12 132 dep pectins 1856:1862 arg1 addition 1814:1821 addition 1814:1821 addition 1814:1821 In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. 28619057 16 133 theme SPE/MAS 2573:2579 arg1 spectroscopy 2586:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 magic-angle spinning (SPE/MAS) NMR spectroscopy 2551:2597 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 9 134 theme smaller 1485:1491 arg1 heteroxylans 1504:1515 heteroxylans 1504:1515 heteroxylans 1504:1515 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 134 theme smaller 1485:1491 arg1 amounts 1493:1499 smaller amounts 1485:1499 smaller amounts of heteroxylans and heteromannans 1485:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 134 theme smaller 1485:1491 arg1 heteromannans 1521:1533 heteromannans 1521:1533 heteromannans 1521:1533 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 19 135 theme collenchyma 3093:3103 arg1 CWs 3105:3107 the collenchyma CWs 3089:3107 the collenchyma CWs from the different segments 3089:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 9 136 theme abundant 1275:1282 arg1 homogalacturonan 1305:1320 the most abundant polysaccharide [with homogalacturonan 1266:1320 the most abundant polysaccharide [with homogalacturonan (HG) 1266:1325 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 136 theme abundant 1275:1282 arg1 pectin 1255:1260 pectin 1255:1260 pectin 1255:1260 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 136 theme abundant 1275:1282 arg1 HG 1323:1324 HG 1323:1324 HG 1323:1324 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 137 dep abundant 1332:1339 arg1 homogalacturonan 1305:1320 the most abundant polysaccharide [with homogalacturonan 1266:1320 the most abundant polysaccharide [with homogalacturonan (HG) 1266:1325 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 137 dep abundant 1332:1339 arg1 pectin 1255:1260 pectin 1255:1260 pectin 1255:1260 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 137 dep abundant 1332:1339 arg1 HG 1323:1324 HG 1323:1324 HG 1323:1324 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 17 138 theme most 2842:2845 arg1 CWs 2872:2874 most eudicotyledon parenchyma CWs 2842:2874 most eudicotyledon parenchyma CWs 2842:2874 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 0 139 theme collenchyma 31:41 arg1 walls 48:52 collenchyma cell walls 31:52 collenchyma cell walls 31:52 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 9 140 theme [with 1299:1303 arg1 homogalacturonan 1305:1320 the most abundant polysaccharide [with homogalacturonan 1266:1320 the most abundant polysaccharide [with homogalacturonan (HG) 1266:1325 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 140 theme [with 1299:1303 arg1 pectin 1255:1260 pectin 1255:1260 pectin 1255:1260 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 9 140 theme [with 1299:1303 arg1 HG 1323:1324 HG 1323:1324 HG 1323:1324 Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. 28619057 0 141 dep Apium 67:71 arg1 L. 84:85 Apium graveolens L. 67:85 Apium graveolens L. 67:85 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 16 142 theme arabinan 2668:2675 arg1 presence 2656:2663 the presence 2652:2663 the presence 2652:2663 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 3 143 theme collenchyma 525:535 arg1 polysaccharides 495:509 cell wall polysaccharides 485:509 cell wall polysaccharides of peripheral collenchyma 485:535 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 0 144 theme walls 48:52 arg1 compositions 15:26 Polysaccharide compositions 0:26 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles 0:95 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 18 145 theme XG 2926:2927 arg1 content 2929:2935 much higher XG content 2914:2935 much higher XG content 2914:2935 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 14 146 attach present 2297:2303 arg2 those 2291:2295 those 2291:2295 those 2291:2295 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 14 146 attach present 2297:2303 arg1 CWs 2338:2340 most eudicotyledon parenchyma CWs 2308:2340 most eudicotyledon parenchyma CWs 2308:2340 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 16 147 theme presence 2656:2663 arg1 structure 2691:2699 the detailed structure 2678:2699 the detailed structure 2678:2699 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 28619057 17 148 theme polysaccharide 2811:2824 arg1 compositions 2826:2837 similar polysaccharide compositions 2803:2837 similar polysaccharide compositions 2803:2837 CONCLUSIONS Celery collenchyma CWs have similar polysaccharide compositions to most eudicotyledon parenchyma CWs. 28619057 6 149 dep Mature 873:878 arg1 elongated 887:895 elongated 887:895 elongated 887:895 Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. 28619057 19 150 theme methyl 2979:2984 arg1 esterification 2986:2999 methyl esterification 2979:2999 methyl esterification of pectin 2979:3009 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 2 151 theme chemical 406:413 arg1 analysis 415:422 no detailed chemical analysis 394:422 no detailed chemical analysis 394:422 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 0 152 theme celery 59:64 arg1 petioles 88:95 celery (Apium graveolens L.) petioles 59:95 celery (Apium graveolens L.) petioles 59:95 Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. 28619057 2 153 theme parenchyma 372:381 arg1 walls 383:387 eudicotyledon parenchyma walls 358:387 eudicotyledon parenchyma walls 358:387 Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. 28619057 13 154 from xyloglucan 2084:2093 arg1 rich 2076:2079 rich 2076:2079 rich 2076:2079 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 154 from xyloglucan 2084:2093 arg1 fraction 2063:2070 the 4 M KOH fraction 2051:2070 the 4 M KOH fraction 2051:2070 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 3 155 theme polysaccharides 495:509 arg1 compositions 454:465 compositions 454:465 compositions 454:465 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 3 155 theme polysaccharides 495:509 arg1 structures 471:480 structures 471:480 structures 471:480 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 18 156 theme parenchyma 2949:2958 arg1 CWs 2960:2962 celery parenchyma CWs 2942:2962 celery parenchyma CWs 2942:2962 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 11 157 contain had 1741:1743 arg1 pectin 1702:1707 the pectin 1698:1707 the pectin in the CWs of the upper segment 1698:1739 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 11 157 contain had 1741:1743 arg2 degree 1754:1759 a higher degree 1745:1759 a higher degree of methyl esterification 1745:1784 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 13 158 from rich 2076:2079 arg1 heteromannans 2099:2111 heteromannans 2099:2111 heteromannans 2099:2111 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 13 158 from rich 2076:2079 arg1 xyloglucan 2084:2093 xyloglucan 2084:2093 xyloglucan 2084:2093 The CDTA and Na2CO3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and heteromannans, and cellulose was predominant in the final residue. 28619057 14 159 theme eudicotyledon 2313:2325 arg1 CWs 2338:2340 most eudicotyledon parenchyma CWs 2308:2340 most eudicotyledon parenchyma CWs 2308:2340 The structures of the xyloglucans, heteroxylans and heteromannans were deduced from the linkage analysis and were similar to those present in most eudicotyledon parenchyma CWs. 28619057 3 160 theme cell 485:488 arg1 polysaccharides 495:509 cell wall polysaccharides 485:509 cell wall polysaccharides of peripheral collenchyma 485:535 In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. 28619057 18 161 theme celery 2886:2891 arg1 CWs 2905:2907 celery collenchyma CWs 2886:2907 celery collenchyma CWs 2886:2907 However, celery collenchyma CWs have much higher XG content than celery parenchyma CWs. 28619057 19 162 from segments 3128:3135 arg1 variation 3076:3084 some variation 3071:3084 some variation in the collenchyma CWs from the different segments 3071:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 19 162 from segments 3128:3135 arg1 CWs 3105:3107 the collenchyma CWs 3089:3107 the collenchyma CWs from the different segments 3089:3135 The degree of methyl esterification of pectin and the structures of the arabinan side chains of RG-I show some variation in the collenchyma CWs from the different segments. 28619057 4 163 theme wall 638:641 arg1 composition 643:653 the cell wall composition 629:653 the cell wall composition of collenchyma from any plant 629:683 RESULTS This is the first detailed investigation of the cell wall composition of collenchyma from any plant. 28619057 11 164 theme other 1795:1799 arg1 segments 1801:1808 the other segments 1791:1808 the other segments 1791:1808 Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. 28619057 15 165 theme semi-rigid 2461:2470 arg1 polysaccharides 2472:2486 the rigid and semi-rigid polysaccharides 2447:2486 the rigid and semi-rigid polysaccharides 2447:2486 Cross polarization with magic angle spinning (CP/MAS) NMR spectroscopy showed no apparent difference in the rigid and semi-rigid polysaccharides in the CWs of the three segments. 28619057 16 166 theme Single-pulse 2522:2533 arg1 excitation 2535:2544 Single-pulse excitation 2522:2544 Single-pulse excitation 2522:2544 Single-pulse excitation with magic-angle spinning (SPE/MAS) NMR spectroscopy, which detects highly mobile polysaccharides, showed the presence of arabinan, the detailed structure of which varied among the cell walls from the three segments. 26963691 6 0 theme whole 894:898 arg1 cycle 910:914 the whole catalytic cycle 890:914 the whole catalytic cycle 890:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 1 1 theme metabolic 147:155 arg1 essential 168:176 essential 168:176 essential 168:176 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 1 1 theme metabolic 147:155 arg1 process 157:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 6 2 theme cycle 910:914 arg1 glycosylation 830:842 the glycosylation 826:842 the glycosylation of the substrate 826:859 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 2 theme cycle 910:914 arg1 step 882:885 the rate limiting step 864:885 the rate limiting step of the whole catalytic cycle 864:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 4 3 theme glycoside 586:594 arg1 hydrolases 596:605 glycoside hydrolases 586:605 glycoside phosphorylases rather than glycoside hydrolases 549:605 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 9 4 theme key 1621:1623 arg1 role 1625:1628 a key role 1619:1628 a key role 1619:1628 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 8 5 theme NagZ 1368:1371 arg1 enzymes 1373:1379 NagZ enzymes 1368:1379 NagZ enzymes 1368:1379 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 9 6 theme glycosylation 1394:1406 arg1 steps 1428:1432 both the glycosylation and deglycosylation steps 1385:1432 steps 1428:1432 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 10 7 theme enzymes 1742:1748 arg1 activity 1725:1732 the phosphorylase activity 1707:1732 the phosphorylase activity of NagZ enzymes 1707:1748 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 4 8 theme experimental 499:510 arg1 study 512:516 a recent experimental study 490:516 a recent experimental study 490:516 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 5 9 theme atomistic 668:676 arg1 level 678:682 the atomistic level 664:682 the atomistic level 664:682 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 6 10 theme calculation 796:806 arg1 results 808:814 Our calculation results 792:814 Our calculation results 792:814 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 0 11 dep study 117:121 arg1 Insights 0:7 Insights 0:7 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis 0:106 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 9 12 theme substrate 1508:1516 arg1 distortion 1518:1527 the substrate distortion 1504:1527 the substrate distortion in the active site 1504:1546 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 3 13 theme secondary 393:401 arg1 structures 403:412 secondary structures 393:412 secondary structures 393:412 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 10 14 theme first 1662:1666 arg1 report 1682:1687 the first computational report 1658:1687 the first computational report for understanding the phosphorylase activity of NagZ enzymes 1658:1748 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 10 14 theme first 1662:1666 arg1 This 1650:1653 This 1650:1653 This 1650:1653 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 6 15 with step 882:885 arg1 barrier 931:937 an energy barrier 921:937 an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 921:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 16 theme mol 952:954 arg1 barrier 931:937 an energy barrier 921:937 an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 921:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 17 theme limiting 873:880 arg1 glycosylation 830:842 the glycosylation 826:842 the glycosylation of the substrate 826:859 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 17 theme limiting 873:880 arg1 step 882:885 the rate limiting step 864:885 the rate limiting step of the whole catalytic cycle 864:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 5 18 theme catalytic 634:642 arg1 process 644:650 the catalytic process 630:650 the catalytic process of NagZs 630:659 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 6 19 theme free 983:986 arg1 mol 952:954 19.3 kcal mol 942:954 19.3 kcal mol(-1) 942:958 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 19 theme free 983:986 arg1 barrier 995:1001 the free energy barrier 979:1001 the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 979:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 8 20 theme enzymes 1373:1379 arg1 activity 1356:1363 the previously suggested phosphorylase activity 1317:1363 the previously suggested phosphorylase activity of NagZ enzymes 1317:1379 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 6 21 gly glycosylation 830:842 arg1 substrate 851:859 the substrate 847:859 the substrate 847:859 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 22 theme kcal 1009:1012 arg1 -1 1018:1019 -1 1018:1019 -1 1018:1019 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 22 theme kcal 1009:1012 arg1 mol 1014:1016 16.4 kcal mol 1004:1016 16.4 kcal mol(-1) 1004:1020 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 0 23 theme QM/MM 111:115 arg1 study 117:121 a QM/MM study 109:121 a QM/MM study 109:121 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 6 24 theme rate 868:871 arg1 glycosylation 830:842 the glycosylation 826:842 the glycosylation of the substrate 826:859 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 24 theme rate 868:871 arg1 step 882:885 the rate limiting step 864:885 the rate limiting step of the whole catalytic cycle 864:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 9 25 theme deglycosylation 1412:1426 arg1 steps 1428:1432 both the glycosylation and deglycosylation steps 1385:1432 steps 1428:1432 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 9 26 theme oxocarbenium-ion-like 1439:1459 arg1 states 1472:1477 the oxocarbenium-ion-like transition states 1435:1477 the oxocarbenium-ion-like transition states 1435:1477 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 6 27 theme kcal 947:950 arg1 -1 956:957 -1 956:957 -1 956:957 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 27 theme kcal 947:950 arg1 mol 952:954 19.3 kcal mol 942:954 19.3 kcal mol(-1) 942:958 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 27 theme kcal 947:950 arg1 barrier 995:1001 the free energy barrier 979:1001 the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 979:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 2 28 theme terminal 306:313 arg1 N-acetylglucosamine 315:333 terminal N-acetylglucosamine 306:333 terminal N-acetylglucosamine 306:333 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 7 29 dep hydrolysis 1100:1109 arg1 the 1096:1098 the 1096:1098 the 1096:1098 For deglycosylation, both the hydrolysis and phosphorylation of the glycosyl-enzyme intermediate were explored. 26963691 4 30 theme glycoside 549:557 arg1 enzymes 537:543 NagZ enzymes 532:543 NagZ enzymes 532:543 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 4 30 theme glycoside 549:557 arg1 phosphorylases 559:572 glycoside phosphorylases 549:572 glycoside phosphorylases rather than glycoside hydrolases 549:605 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 5 31 theme NagZs 655:659 arg1 process 644:650 the catalytic process 630:650 the catalytic process of NagZs 630:659 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 6 32 dep barrier 995:1001 arg1 -1 1018:1019 -1 1018:1019 -1 1018:1019 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 32 dep barrier 995:1001 arg1 mol 1014:1016 16.4 kcal mol 1004:1016 16.4 kcal mol(-1) 1004:1020 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 1 33 theme bacterial 182:190 arg1 growth 192:197 bacterial growth 182:197 bacterial growth 182:197 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 0 34 theme catalytic 18:26 arg1 mechanism 28:36 the catalytic mechanism 14:36 the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase 14:83 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 10 35 theme computational 1668:1680 arg1 report 1682:1687 the first computational report 1658:1687 the first computational report for understanding the phosphorylase activity of NagZ enzymes 1658:1748 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 10 35 theme computational 1668:1680 arg1 This 1650:1653 This 1650:1653 This 1650:1653 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 9 36 theme active 1536:1541 arg1 site 1543:1546 the active site 1532:1546 the active site 1532:1546 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 5 37 theme QM/MM 776:780 arg1 approach 782:789 a QM/MM approach 774:789 a QM/MM approach 774:789 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 10 38 theme NagZ 1737:1740 arg1 enzymes 1742:1748 NagZ enzymes 1737:1748 NagZ enzymes 1737:1748 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 3 39 theme hydrolase 459:467 arg1 GH3 476:478 the glycoside hydrolase family GH3 445:478 the glycoside hydrolase family GH3 445:478 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 3 40 theme acid 375:378 arg1 sequence 380:387 the amino acid sequence 365:387 the amino acid sequence 365:387 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 5 41 theme NagZ 711:714 arg1 mechanism 698:706 the reaction mechanism 685:706 the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) 685:746 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 0 42 from subtilis 99:106 arg1 Insights 0:7 Insights 0:7 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis 0:106 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 6 43 theme experimental 1042:1053 arg1 constant 1060:1067 the experimental rate constant 1038:1067 the experimental rate constant 1038:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 9 44 theme transition 1461:1470 arg1 states 1472:1477 the oxocarbenium-ion-like transition states 1435:1477 the oxocarbenium-ion-like transition states 1435:1477 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 3 45 theme family 469:474 arg1 GH3 476:478 the glycoside hydrolase family GH3 445:478 the glycoside hydrolase family GH3 445:478 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 2 46 attach removal 295:301 arg2 N-acetylglucosamine 315:333 terminal N-acetylglucosamine 306:333 terminal N-acetylglucosamine 306:333 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 2 46 attach removal 295:301 arg3 fragments 341:349 PG fragments 338:349 PG fragments 338:349 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 10 47 theme phosphorylase 1711:1723 arg1 activity 1725:1732 the phosphorylase activity 1707:1732 the phosphorylase activity of NagZ enzymes 1707:1748 This is the first computational report for understanding the phosphorylase activity of NagZ enzymes. 26963691 6 48 theme substrate 851:859 arg1 step 882:885 the rate limiting step 864:885 the rate limiting step of the whole catalytic cycle 864:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 48 theme substrate 851:859 arg1 glycosylation 830:842 the glycosylation 826:842 the glycosylation of the substrate 826:859 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 8 49 theme theoretical 1294:1304 arg1 support 1306:1312 theoretical support 1294:1312 theoretical support 1294:1312 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 3 50 theme NagZ 415:418 arg1 enzymes 420:426 NagZ enzymes 415:426 NagZ enzymes 415:426 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 6 51 theme energy 924:929 arg1 barrier 931:937 an energy barrier 921:937 an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 921:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 5 52 from subtilis 730:737 arg1 NagZ 711:714 NagZ 711:714 NagZ from Bacillus subtilis (BsNagZ) 711:746 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 5 52 from subtilis 730:737 arg1 mechanism 698:706 the reaction mechanism 685:706 the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) 685:746 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 3 53 theme amino 369:373 arg1 sequence 380:387 the amino acid sequence 365:387 the amino acid sequence 365:387 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 3 54 theme glycoside 449:457 arg1 GH3 476:478 the glycoside hydrolase family GH3 445:478 the glycoside hydrolase family GH3 445:478 According to the amino acid sequence and secondary structures, NagZ enzymes should belong to the glycoside hydrolase family GH3. 26963691 0 55 theme glycoside 65:73 arg1 hydrolase 75:83 N-acetylglucosaminidase glycoside hydrolase 41:83 N-acetylglucosaminidase glycoside hydrolase 41:83 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 7 56 theme intermediate 1154:1165 arg1 phosphorylation 1115:1129 phosphorylation 1115:1129 phosphorylation 1115:1129 For deglycosylation, both the hydrolysis and phosphorylation of the glycosyl-enzyme intermediate were explored. 26963691 7 56 theme intermediate 1154:1165 arg1 hydrolysis 1100:1109 hydrolysis 1100:1109 hydrolysis 1100:1109 For deglycosylation, both the hydrolysis and phosphorylation of the glycosyl-enzyme intermediate were explored. 26963691 8 57 theme energy 1227:1232 arg1 barrier 1234:1240 the lower energy barrier 1217:1240 the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)) 1217:1284 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 0 58 theme N-acetylglucosaminidase 41:63 arg1 hydrolase 75:83 N-acetylglucosaminidase glycoside hydrolase 41:83 N-acetylglucosaminidase glycoside hydrolase 41:83 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 6 59 theme catalytic 900:908 arg1 cycle 910:914 the whole catalytic cycle 890:914 the whole catalytic cycle 890:914 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 2 60 theme NagZ 228:231 arg1 β-N-Acetylglucosaminidases 200:225 β-N-Acetylglucosaminidases 200:225 β-N-Acetylglucosaminidases (NagZ enzymes) 200:240 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 2 60 theme NagZ 228:231 arg1 enzymes 233:239 NagZ enzymes 228:239 NagZ enzymes 228:239 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 2 61 theme PG 338:339 arg1 fragments 341:349 PG fragments 338:349 PG fragments 338:349 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 4 62 theme NagZ 532:535 arg1 enzymes 537:543 NagZ enzymes 532:543 NagZ enzymes 532:543 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 4 62 theme NagZ 532:535 arg1 phosphorylases 559:572 glycoside phosphorylases 549:572 glycoside phosphorylases rather than glycoside hydrolases 549:605 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 5 63 theme reaction 689:696 arg1 mechanism 698:706 the reaction mechanism 685:706 the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) 685:746 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 2 64 theme PG 262:263 arg1 process 265:271 the PG process 258:271 the PG process 258:271 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 1 65 theme peptidoglycan 128:140 arg1 essential 168:176 essential 168:176 essential 168:176 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 1 65 theme peptidoglycan 128:140 arg1 process 157:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 6 66 theme rate 1055:1058 arg1 constant 1060:1067 the experimental rate constant 1038:1067 the experimental rate constant 1038:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 8 67 theme kcal 1272:1275 arg1 mol 1277:1279 1.8 vs. 17.7 kcal mol(-1) 1259:1283 1.8 vs. 17.7 kcal mol(-1) 1259:1283 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 8 67 theme kcal 1272:1275 arg1 hydrolysis 1247:1256 hydrolysis 1247:1256 hydrolysis (1.8 vs. 17.7 kcal mol(-1)) 1247:1284 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 2 68 theme N-acetylglucosamine 315:333 arg1 removal 295:301 the removal 291:301 the removal of terminal N-acetylglucosamine in PG fragments 291:349 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 26963691 5 69 from mechanism 698:706 arg1 BsNagZ 740:745 BsNagZ 740:745 BsNagZ 740:745 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 5 69 from mechanism 698:706 arg1 subtilis 730:737 Bacillus subtilis 721:737 Bacillus subtilis (BsNagZ) 721:746 To further understand the catalytic process of NagZs at the atomistic level, the reaction mechanism of NagZ from Bacillus subtilis (BsNagZ) has been studied by using a QM/MM approach. 26963691 7 70 theme glycosyl-enzyme 1138:1152 arg1 intermediate 1154:1165 the glycosyl-enzyme intermediate 1134:1165 the glycosyl-enzyme intermediate 1134:1165 For deglycosylation, both the hydrolysis and phosphorylation of the glycosyl-enzyme intermediate were explored. 26963691 8 71 theme lower 1221:1225 arg1 barrier 1234:1240 the lower energy barrier 1217:1240 the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)) 1217:1284 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 8 72 theme phosphorylase 1342:1354 arg1 activity 1356:1363 the previously suggested phosphorylase activity 1317:1363 the previously suggested phosphorylase activity of NagZ enzymes 1317:1379 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 6 73 theme energy 988:993 arg1 mol 952:954 19.3 kcal mol 942:954 19.3 kcal mol(-1) 942:958 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 6 73 theme energy 988:993 arg1 barrier 995:1001 the free energy barrier 979:1001 the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant 979:1067 Our calculation results show that the glycosylation of the substrate is the rate limiting step of the whole catalytic cycle with an energy barrier of 19.3 kcal mol(-1), which is close to the free energy barrier (16.4 kcal mol(-1)) estimated from the experimental rate constant. 26963691 1 74 theme PG 143:144 arg1 essential 168:176 essential 168:176 essential 168:176 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 1 74 theme PG 143:144 arg1 process 157:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process 124:163 The peptidoglycan (PG) metabolic process is essential for bacterial growth. 26963691 0 75 theme hydrolase 75:83 arg1 mechanism 28:36 the catalytic mechanism 14:36 the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase 14:83 Insights into the catalytic mechanism of N-acetylglucosaminidase glycoside hydrolase from Bacillus subtilis: a QM/MM study. 26963691 4 76 theme recent 492:497 arg1 study 512:516 a recent experimental study 490:516 a recent experimental study 490:516 However, a recent experimental study revealed that NagZ enzymes are glycoside phosphorylases rather than glycoside hydrolases. 26963691 8 77 theme suggested 1332:1340 arg1 activity 1356:1363 the previously suggested phosphorylase activity 1317:1363 the previously suggested phosphorylase activity of NagZ enzymes 1317:1379 The phosphorylation corresponds to the lower energy barrier than hydrolysis (1.8 vs. 17.7 kcal mol(-1)), giving theoretical support to the previously suggested phosphorylase activity of NagZ enzymes. 26963691 9 78 from distortion 1518:1527 arg1 site 1543:1546 the active site 1532:1546 the active site 1532:1546 In both the glycosylation and deglycosylation steps, the oxocarbenium-ion-like transition states are always involved, and the substrate distortion in the active site can significantly facilitate the reaction, in which residue D123 plays a key role in this distortion. 26963691 2 79 from removal 295:301 arg1 fragments 341:349 PG fragments 338:349 PG fragments 338:349 β-N-Acetylglucosaminidases (NagZ enzymes) are involved in the PG process and they catalyze the removal of terminal N-acetylglucosamine in PG fragments. 27077820 1 0 theme oligosaccharide 255:269 arg1 synthesis 271:279 oligosaccharide synthesis 255:279 oligosaccharide synthesis 255:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 2 1 theme per-O-trimethylsilylated 298:321 arg1 donor 363:367 the glycosyl donor 350:367 the glycosyl donor 350:367 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 2 1 theme per-O-trimethylsilylated 298:321 arg1 iodides 332:338 the per-O-trimethylsilylated glycosyl iodides 294:338 the per-O-trimethylsilylated glycosyl iodides 294:338 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 3 2 theme effective 576:584 arg1 trialkylamine 528:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 3 2 theme effective 576:584 arg1 promoter 586:593 an effective promoter 573:593 an effective promoter for the glycosylation 573:615 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 1 3 theme Trimethylsilyl 93:106 arg1 TMS 109:111 TMS 109:111 TMS 109:111 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 1 3 theme Trimethylsilyl 93:106 arg1 group 125:129 Trimethylsilyl (TMS) protecting group 93:129 Trimethylsilyl (TMS) protecting group 93:129 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 4 4 dep yields 698:703 arg1 % 696:696 % 696:696 % 696:696 Importantly, the reaction was α-stereospecific and gave the products in 58%-78% yields. 27077820 1 5 theme simultaneous 172:183 arg1 protection 185:194 the simultaneous protection 168:194 the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis 168:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 2 6 theme hydroxyl 412:419 arg1 groups 421:426 selectively exposed primary hydroxyl groups 384:426 selectively exposed primary hydroxyl groups 384:426 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 2 7 theme primary 404:410 arg1 groups 421:426 selectively exposed primary hydroxyl groups 384:426 selectively exposed primary hydroxyl groups 384:426 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 1 8 theme protecting 114:123 arg1 TMS 109:111 TMS 109:111 TMS 109:111 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 1 8 theme protecting 114:123 arg1 group 125:129 Trimethylsilyl (TMS) protecting group 93:129 Trimethylsilyl (TMS) protecting group 93:129 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 4 9 from products 678:685 arg1 yields 698:703 58%-78% yields 690:703 58%-78% yields 690:703 Importantly, the reaction was α-stereospecific and gave the products in 58%-78% yields. 27077820 2 10 theme glycosyl 455:462 arg1 acceptor 464:471 the glycosyl acceptor 451:471 the glycosyl acceptor for the reaction 451:488 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 2 11 theme glycosyl 323:330 arg1 donor 363:367 the glycosyl donor 350:367 the glycosyl donor 350:367 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 2 11 theme glycosyl 323:330 arg1 iodides 332:338 the per-O-trimethylsilylated glycosyl iodides 294:338 the per-O-trimethylsilylated glycosyl iodides 294:338 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 3 12 theme cheap 495:499 arg1 triethylamine 543:555 triethylamine 543:555 triethylamine 543:555 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 3 12 theme cheap 495:499 arg1 trialkylamine 528:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 3 12 theme cheap 495:499 arg1 promoter 586:593 an effective promoter 573:593 an effective promoter for the glycosylation 573:615 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 4 13 theme -78 693:695 arg1 % 692:692 % 692:692 % 692:692 Importantly, the reaction was α-stereospecific and gave the products in 58%-78% yields. 27077820 1 14 theme glycosyl 208:215 arg1 donor- 217:222 the glycosyl donor- 204:222 the glycosyl donor- 204:222 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 1 15 from synthesis 271:279 arg1 protection 185:194 the simultaneous protection 168:194 the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis 168:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 1 16 theme donor- 217:222 arg1 protection 185:194 the simultaneous protection 168:194 the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis 168:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 0 17 theme glycosyl 77:84 arg1 donor 86:90 glycosyl donor 77:90 glycosyl donor 77:90 Synthesis of oligosaccharides using per-O-trimethylsilyl-glycosyl iodides as glycosyl donor. 27077820 1 18 from acceptor-substrates 232:250 arg1 synthesis 271:279 oligosaccharide synthesis 255:279 oligosaccharide synthesis 255:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 1 19 from donor- 217:222 arg1 synthesis 271:279 oligosaccharide synthesis 255:279 oligosaccharide synthesis 255:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 3 20 theme available 518:526 arg1 triethylamine 543:555 triethylamine 543:555 triethylamine 543:555 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 3 20 theme available 518:526 arg1 trialkylamine 528:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine 491:540 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 3 20 theme available 518:526 arg1 promoter 586:593 an effective promoter 573:593 an effective promoter for the glycosylation 573:615 The cheap and commercially available trialkylamine, triethylamine was found to be an effective promoter for the glycosylation. 27077820 4 21 dep % 696:696 arg1 % 692:692 % 692:692 % 692:692 Importantly, the reaction was α-stereospecific and gave the products in 58%-78% yields. 27077820 2 22 theme glycosyl 354:361 arg1 donor 363:367 the glycosyl donor 350:367 the glycosyl donor 350:367 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 2 22 theme glycosyl 354:361 arg1 iodides 332:338 the per-O-trimethylsilylated glycosyl iodides 294:338 the per-O-trimethylsilylated glycosyl iodides 294:338 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 1 23 from protection 185:194 arg1 synthesis 271:279 oligosaccharide synthesis 255:279 oligosaccharide synthesis 255:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 2 24 theme exposed 396:402 arg1 groups 421:426 selectively exposed primary hydroxyl groups 384:426 selectively exposed primary hydroxyl groups 384:426 Thus, while the per-O-trimethylsilylated glycosyl iodides served as the glycosyl donor, those bearing selectively exposed primary hydroxyl groups were found suitable as the glycosyl acceptor for the reaction. 27077820 4 25 theme 58 690:691 arg1 % 692:692 % 692:692 % 692:692 Importantly, the reaction was α-stereospecific and gave the products in 58%-78% yields. 27077820 1 26 theme acceptor-substrates 232:250 arg1 protection 185:194 the simultaneous protection 168:194 the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis 168:279 Trimethylsilyl (TMS) protecting group has been found to be very useful for the simultaneous protection of both the glycosyl donor- and the acceptor-substrates in oligosaccharide synthesis. 27077820 0 27 theme oligosaccharides 13:28 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of oligosaccharides using per-O-trimethylsilyl-glycosyl 0:64 Synthesis of oligosaccharides using per-O-trimethylsilyl-glycosyl iodides as glycosyl donor. 27714557 0 0 theme sugar 120:124 arg1 oxazoline 126:134 Coprinopsis cinerea endoglycosidase and sugar oxazoline 80:134 oxazoline 126:134 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 4 1 theme GlcNAc-RNase 805:816 arg1 B 818:818 80 μg GlcNAc-RNase B 799:818 80 μg GlcNAc-RNase B 799:818 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 5 2 theme transglycosylation 935:952 arg1 method 954:959 This transglycosylation method 930:959 This transglycosylation method using SG-oxazoline and Endo-CCN180H 930:995 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 1 3 theme modification 199:210 arg1 method 174:179 an efficient method 161:179 an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities 161:340 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 4 4 theme reaction 654:661 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 4 theme reaction 654:661 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 5 5 theme pharmaceutical 1025:1038 arg1 glycoproteins 1040:1052 pharmaceutical glycoproteins 1025:1052 pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides 1025:1119 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 4 6 theme 80 μg 799:803 arg1 B 818:818 80 μg GlcNAc-RNase B 799:818 80 μg GlcNAc-RNase B 799:818 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 7 theme efficient 644:652 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 7 theme efficient 644:652 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 2 8 theme GlcNAc-RNase 375:386 arg1 B 388:388 GlcNAc-RNase B 375:388 GlcNAc-RNase B; acceptor 375:398 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 4 9 theme complex 768:774 arg1 oligosaccharide 776:790 sialo-biantennary-type complex oligosaccharide 745:790 sialo-biantennary-type complex oligosaccharide 745:790 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 10 theme 20 μl 866:870 arg1 pH 887:888 20 μl 20 mM Tris/HCl pH 7.5 866:892 20 μl 20 mM Tris/HCl pH 7.5 866:892 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 3 11 theme transglycosylation 606:623 arg1 reaction 625:632 Endo-CC's transglycosylation reaction 596:632 Endo-CC's transglycosylation reaction 596:632 pH 7.5 was ideal for both SG-oxazoline's stability and Endo-CC's transglycosylation reaction. 27714557 5 12 theme biantennary-complex-type 1079:1102 arg1 oligosaccharides 1104:1119 homogenous biantennary-complex-type oligosaccharides 1068:1119 homogenous biantennary-complex-type oligosaccharides 1068:1119 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 1 13 gly glycoproteins 258:270 arg1 glycoproteins 258:270 glycoproteins 258:270 glycoproteins structurally homogeneous 258:295 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 2 14 theme Coprinopsis 460:470 arg1 endo-β-N-acetylglucosaminidase 480:509 Coprinopsis cinerea endo-β-N-acetylglucosaminidase 460:509 Coprinopsis cinerea endo-β-N-acetylglucosaminidase 460:509 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 15 dep RESULTS 343:349 arg1 B 372:372 Deglycosylated-RNase B 351:372 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 16 theme N180H 444:448 arg1 Endo-CCN180H 512:523 Endo-CCN180H 512:523 Endo-CCN180H 512:523 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 16 theme N180H 444:448 arg1 mutant 450:455 an N180H mutant 441:455 an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) 441:524 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 16 theme N180H 444:448 arg1 RESULTS 343:349 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 17 theme Deglycosylated-RNase 351:370 arg1 B 372:372 Deglycosylated-RNase B 351:372 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 1 18 theme N-linked 223:230 arg1 chains 248:253 N-linked oligosaccharide chains 223:253 N-linked oligosaccharide chains 223:253 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 0 19 theme efficient 7:15 arg1 transglycosylation 17:34 Highly efficient transglycosylation 0:34 Highly efficient transglycosylation of sialo-complex-type oligosaccharide 0:72 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 4 20 from Endo-CCN180H 850:861 arg1 pH 887:888 20 μl 20 mM Tris/HCl pH 7.5 866:892 20 μl 20 mM Tris/HCl pH 7.5 866:892 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 21 theme 3 μg 845:848 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 21 theme 3 μg 845:848 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 1 22 theme oligosaccharide 232:246 arg1 chains 248:253 N-linked oligosaccharide chains 223:253 N-linked oligosaccharide chains 223:253 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 1 23 link N-linked 223:230 arg1 chains 248:253 N-linked oligosaccharide chains 223:253 N-linked oligosaccharide chains 223:253 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 4 24 theme 20 mM 872:876 arg1 pH 887:888 20 μl 20 mM Tris/HCl pH 7.5 866:892 20 μl 20 mM Tris/HCl pH 7.5 866:892 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 2 25 theme cinerea 472:478 arg1 endo-β-N-acetylglucosaminidase 480:509 Coprinopsis cinerea endo-β-N-acetylglucosaminidase 460:509 Coprinopsis cinerea endo-β-N-acetylglucosaminidase 460:509 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 26 theme endo-β-N-acetylglucosaminidase 480:509 arg1 Endo-CCN180H 512:523 Endo-CCN180H 512:523 Endo-CCN180H 512:523 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 26 theme endo-β-N-acetylglucosaminidase 480:509 arg1 -oxazoline 418:427 sialylglyco (SG)-oxazoline 402:427 sialylglyco (SG)-oxazoline (donor) 402:435 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 26 theme endo-β-N-acetylglucosaminidase 480:509 arg1 RESULTS 343:349 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 26 theme endo-β-N-acetylglucosaminidase 480:509 arg1 mutant 450:455 an N180H mutant 441:455 an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) 441:524 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 26 theme endo-β-N-acetylglucosaminidase 480:509 arg1 donor 430:434 donor 430:434 donor 430:434 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 5 27 theme homogenous 1068:1077 arg1 oligosaccharides 1104:1119 homogenous biantennary-complex-type oligosaccharides 1068:1119 homogenous biantennary-complex-type oligosaccharides 1068:1119 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 5 28 dep CONCLUSIONS 918:928 arg1 beneficial 1000:1009 beneficial 1000:1009 beneficial 1000:1009 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 1 29 theme glycoproteins 258:270 arg1 chains 248:253 N-linked oligosaccharide chains 223:253 N-linked oligosaccharide chains 223:253 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 0 30 theme oligosaccharide 58:72 arg1 transglycosylation 17:34 Highly efficient transglycosylation 0:34 Highly efficient transglycosylation of sialo-complex-type oligosaccharide 0:72 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 4 31 theme B 818:818 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 31 theme B 818:818 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 0 32 theme sialo-complex-type 39:56 arg1 oligosaccharide 58:72 sialo-complex-type oligosaccharide 39:72 sialo-complex-type oligosaccharide 39:72 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 1 33 theme homogeneous 285:295 arg1 glycoproteins 258:270 glycoproteins 258:270 glycoproteins structurally homogeneous 258:295 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 0 34 theme Coprinopsis 80:90 arg1 endoglycosidase 100:114 Coprinopsis cinerea endoglycosidase and sugar oxazoline 80:134 endoglycosidase 100:114 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 0 35 gly transglycosylation 17:34 arg1 oligosaccharide 58:72 sialo-complex-type oligosaccharide 39:72 sialo-complex-type oligosaccharide 39:72 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 2 36 theme SG 415:416 arg1 -oxazoline 418:427 sialylglyco (SG)-oxazoline 402:427 sialylglyco (SG)-oxazoline (donor) 402:435 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 36 theme SG 415:416 arg1 RESULTS 343:349 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 36 theme SG 415:416 arg1 donor 430:434 donor 430:434 donor 430:434 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 37 dep B 388:388 arg1 acceptor 391:398 acceptor 391:398 GlcNAc-RNase B; acceptor 375:398 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 5 38 gly glycoproteins 1040:1052 arg1 glycoproteins 1040:1052 pharmaceutical glycoproteins 1025:1052 pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides 1025:1119 CONCLUSIONS This transglycosylation method using SG-oxazoline and Endo-CCN180H is beneficial for producing pharmaceutical glycoproteins modified with homogenous biantennary-complex-type oligosaccharides. 27714557 1 39 theme efficient 164:172 arg1 method 174:179 an efficient method 161:179 an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities 161:340 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 27714557 4 40 theme glycosylated-RNase 688:705 arg1 B 707:707 glycosylated-RNase B 688:707 glycosylated-RNase B 688:707 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 41 theme Tris/HCl 878:885 arg1 pH 887:888 20 μl 20 mM Tris/HCl pH 7.5 866:892 20 μl 20 mM Tris/HCl pH 7.5 866:892 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 42 theme SG-oxazoline 828:839 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 42 theme SG-oxazoline 828:839 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 43 theme 200 μg 821:826 arg1 SG-oxazoline 828:839 200 μg SG-oxazoline 821:839 200 μg SG-oxazoline 821:839 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 2 44 theme sialylglyco 402:412 arg1 -oxazoline 418:427 sialylglyco (SG)-oxazoline 402:427 sialylglyco (SG)-oxazoline (donor) 402:435 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 44 theme sialylglyco 402:412 arg1 RESULTS 343:349 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor) 343:399 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 2 44 theme sialylglyco 402:412 arg1 donor 430:434 donor 430:434 donor 430:434 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 4 45 from 30 °C 897:901 arg1 Endo-CCN180H 850:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 : 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H 797:861 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 4 45 from 30 °C 897:901 arg1 conditions 663:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions 635:672 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 2 46 dep B 372:372 arg1 B 388:388 GlcNAc-RNase B 375:388 GlcNAc-RNase B; acceptor 375:398 RESULTS Deglycosylated-RNase B (GlcNAc-RNase B; acceptor), sialylglyco (SG)-oxazoline (donor) and an N180H mutant of Coprinopsis cinerea endo-β-N-acetylglucosaminidase (Endo-CCN180H) were employed. 27714557 4 47 theme sialo-biantennary-type 745:766 arg1 oligosaccharide 776:790 sialo-biantennary-type complex oligosaccharide 745:790 sialo-biantennary-type complex oligosaccharide 745:790 The most efficient reaction conditions for producing glycosylated-RNase B, virtually modified completely with sialo-biantennary-type complex oligosaccharide, were: 80 μg GlcNAc-RNase B, 200 μg SG-oxazoline and 3 μg Endo-CCN180H in 20 μl 20 mM Tris/HCl pH 7.5 at 30 °C for 30-60 min. 27714557 0 48 theme cinerea 92:98 arg1 endoglycosidase 100:114 Coprinopsis cinerea endoglycosidase and sugar oxazoline 80:134 endoglycosidase 100:114 Highly efficient transglycosylation of sialo-complex-type oligosaccharide using Coprinopsis cinerea endoglycosidase and sugar oxazoline. 27714557 1 49 theme chemoenzymatic 184:197 arg1 modification 199:210 chemoenzymatic modification 184:210 chemoenzymatic modification 184:210 OBJECTIVES To establish an efficient method of chemoenzymatic modification for making N-linked oligosaccharide chains of glycoproteins structurally homogeneous, which crucially affects their bioactivities. 28853271 2 0 theme glycan 450:455 arg1 profiling 457:465 released glycan profiling 441:465 released glycan profiling 441:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 3 1 contain had 491:493 arg1 Omalizumab 480:489 Omalizumab 480:489 Omalizumab 480:489 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 3 1 contain had 491:493 arg1 CMAB007 468:474 CMAB007 468:474 CMAB007 468:474 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 3 1 contain had 491:493 arg2 structure 512:520 the same primary structure 495:520 the same primary structure 495:520 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 2 2 theme biosimilar 272:281 arg1 characterization 240:255 An in depth characterization 228:255 An in depth characterization of a candidate biosimilar 228:281 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 3 dep approach 318:325 arg1 provides 341:348 provides 341:348 provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling 341:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 4 theme candidate 262:270 arg1 biosimilar 272:281 a candidate biosimilar 260:281 a candidate biosimilar 260:281 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 5 5 from drug 865:868 arg1 China 873:877 China 873:877 China 873:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 1 6 dep Omalizumab 124:133 arg1 Xolair 136:141 Xolair 136:141 Xolair 136:141 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 4 7 theme high 692:695 arg1 types 705:709 high mannose types 692:709 high mannose types 692:709 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 5 8 theme E 843:843 arg1 antibody 856:863 the first humanized anti-immunoglobulin E monoclonal antibody 803:863 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 3 9 theme same 499:502 arg1 structure 512:520 the same primary structure 495:520 the same primary structure 495:520 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 5 10 theme monoclonal 845:854 arg1 antibody 856:863 the first humanized anti-immunoglobulin E monoclonal antibody 803:863 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 1 11 theme humanized 146:154 arg1 antibody 189:196 a humanized anti-immunoglobulin E monoclonal antibody 144:196 Xolair; a humanized anti-immunoglobulin E monoclonal antibody 136:196 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 2 12 theme released 441:448 arg1 profiling 457:465 released glycan profiling 441:465 released glycan profiling 441:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 3 13 theme primary 504:510 arg1 structure 512:520 the same primary structure 495:520 the same primary structure 495:520 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 2 14 theme routine 359:365 arg1 tools 367:371 routine tools 359:371 routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling 359:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 15 theme peptide 420:426 arg1 mapping 428:434 peptide mapping 420:434 peptide mapping 420:434 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 1 16 theme E 176:176 arg1 antibody 189:196 a humanized anti-immunoglobulin E monoclonal antibody 144:196 Xolair; a humanized anti-immunoglobulin E monoclonal antibody 136:196 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 5 17 theme anti-immunoglobulin 823:841 arg1 antibody 856:863 the first humanized anti-immunoglobulin E monoclonal antibody 803:863 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 3 18 theme same 547:550 arg1 content 552:558 almost the same content 536:558 almost the same content of C-terminal lysine variants 536:588 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 1 19 theme monoclonal 178:187 arg1 antibody 189:196 a humanized anti-immunoglobulin E monoclonal antibody 144:196 Xolair; a humanized anti-immunoglobulin E monoclonal antibody 136:196 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 3 20 theme variants 581:588 arg1 content 552:558 almost the same content 536:558 almost the same content of C-terminal lysine variants 536:588 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 2 21 theme intact 395:400 arg1 measurement 407:417 accurate intact mass measurement 386:417 accurate intact mass measurement 386:417 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 22 theme mass 402:405 arg1 measurement 407:417 accurate intact mass measurement 386:417 accurate intact mass measurement 386:417 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 1 23 dep Xolair 136:141 arg1 antibody 189:196 a humanized anti-immunoglobulin E monoclonal antibody 144:196 Xolair; a humanized anti-immunoglobulin E monoclonal antibody 136:196 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 2 24 theme accurate 386:393 arg1 measurement 407:417 accurate intact mass measurement 386:417 accurate intact mass measurement 386:417 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 0 25 theme Omalizumab 45:54 arg1 analysis 33:40 analysis 33:40 analysis 33:40 [Structural characterization and analysis of Omalizumab and its biosimilar CMAB007 by LC-MS]. 28853271 0 25 theme Omalizumab 45:54 arg1 characterization 12:27 characterization 12:27 characterization 12:27 [Structural characterization and analysis of Omalizumab and its biosimilar CMAB007 by LC-MS]. 28853271 0 25 theme Omalizumab 45:54 arg1 CMAB007 75:81 its biosimilar CMAB007 60:81 its biosimilar CMAB007 by LC-MS 60:90 [Structural characterization and analysis of Omalizumab and its biosimilar CMAB007 by LC-MS]. 28853271 5 26 theme first 807:811 arg1 antibody 856:863 the first humanized anti-immunoglobulin E monoclonal antibody 803:863 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 3 27 theme C-terminal 563:572 arg1 variants 581:588 C-terminal lysine variants 563:588 C-terminal lysine variants 563:588 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 5 28 theme antibody 856:863 arg1 drug 865:868 the first humanized anti-immunoglobulin E monoclonal antibody drug 803:868 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 4 29 theme oligosaccharides 618:633 arg1 similar 645:651 similar 645:651 similar 645:651 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 4 29 theme oligosaccharides 618:633 arg1 types 595:599 The types 591:599 The types of detected free oligosaccharides 591:633 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 2 30 theme tools 367:371 arg1 set 352:354 a set 350:354 a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling 350:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 30 theme tools 367:371 arg1 tools 367:371 routine tools 359:371 routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling 359:465 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 5 31 theme similar 752:758 arg1 molecular 760:768 a highly similar molecular 743:768 a highly similar molecular to Omalizumab 743:782 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 5 31 theme similar 752:758 arg1 CMAB007 712:718 CMAB007 712:718 CMAB007 712:718 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 3 32 theme lysine 574:579 arg1 variants 581:588 C-terminal lysine variants 563:588 C-terminal lysine variants 563:588 CMAB007 and Omalizumab had the same primary structure and exhibited almost the same content of C-terminal lysine variants. 28853271 4 33 theme free 613:616 arg1 oligosaccharides 618:633 detected free oligosaccharides 604:633 detected free oligosaccharides 604:633 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 2 34 theme systematic 307:316 arg1 approach 318:325 a systematic approach 305:325 a systematic approach 305:325 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 2 35 theme depth 234:238 arg1 characterization 240:255 An in depth characterization 228:255 An in depth characterization of a candidate biosimilar 228:281 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 1 36 theme Omalizumab 124:133 arg1 similarity 110:119 the similarity 106:119 the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007 106:225 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 4 37 theme mannose 697:703 arg1 types 705:709 high mannose types 692:709 high mannose types 692:709 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 5 38 theme humanized 813:821 arg1 antibody 856:863 the first humanized anti-immunoglobulin E monoclonal antibody 803:863 the first humanized anti-immunoglobulin E monoclonal antibody drug in China 803:877 CMAB007 could be considered as a highly similar molecular to Omalizumab and expected to be the first humanized anti-immunoglobulin E monoclonal antibody drug in China. 28853271 2 39 theme in 231:232 arg1 characterization 240:255 An in depth characterization 228:255 An in depth characterization of a candidate biosimilar 228:281 An in depth characterization of a candidate biosimilar was carried out using a systematic approach, the approach provides a set of routine tools that combine accurate intact mass measurement, peptide mapping, and released glycan profiling. 28853271 4 40 theme detected 604:611 arg1 oligosaccharides 618:633 detected free oligosaccharides 604:633 detected free oligosaccharides 604:633 The types of detected free oligosaccharides were very similar, such as sialylation, fucosylation and high mannose types. 28853271 1 41 theme anti-immunoglobulin 156:174 arg1 antibody 189:196 a humanized anti-immunoglobulin E monoclonal antibody 144:196 Xolair; a humanized anti-immunoglobulin E monoclonal antibody 136:196 We compared the similarity of Omalizumab (Xolair; a humanized anti-immunoglobulin E monoclonal antibody) and it's biosimilar CMAB007. 28853271 0 42 theme biosimilar 64:73 arg1 CMAB007 75:81 its biosimilar CMAB007 60:81 its biosimilar CMAB007 by LC-MS 60:90 [Structural characterization and analysis of Omalizumab and its biosimilar CMAB007 by LC-MS]. 24549353 0 0 theme oligosaccharides 72:87 arg1 synthesis 43:51 the modular synthesis 31:51 the modular synthesis of heparan sulfate oligosaccharides 31:87 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 2 1 theme libraries 488:496 arg1 assembly 476:483 the assembly 472:483 the assembly of libraries of HS oligosaccharides 472:519 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 1 2 theme proteins 140:147 arg1 hundreds 99:106 hundreds 99:106 hundreds of heparan sulfate (HS) binding proteins 99:147 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 3 3 theme temporary 811:819 arg1 Lev 833:835 Lev 833:835 Lev 833:835 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 3 theme temporary 811:819 arg1 levulinoyl 821:830 temporary levulinoyl 811:830 temporary levulinoyl (Lev) 811:836 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 4 theme little 232:237 arg1 information 239:249 very little information 227:249 very little information 227:249 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 0 5 theme sulfate 64:70 arg1 oligosaccharides 72:87 heparan sulfate oligosaccharides 56:87 heparan sulfate oligosaccharides 56:87 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 5 6 theme favorable 1124:1132 arg1 properties 1152:1161 more favorable glycosyl donating properties 1119:1161 more favorable glycosyl donating properties 1119:1161 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 5 7 theme C-4 1240:1242 arg1 hydroxyl 1244:1251 the C-4 hydroxyl 1236:1251 the C-4 hydroxyl 1236:1251 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 2 8 theme relationship 544:555 arg1 studies 557:563 structure-activity relationship studies 525:563 structure-activity relationship studies 525:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 3 9 theme coupling 696:703 arg1 products 705:712 coupling products 696:712 coupling products 696:712 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 4 10 theme protecting 1040:1049 arg1 group 1051:1055 an ideal protecting group 1031:1055 it an ideal protecting group for HS oligosaccharide assembly 1028:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 2 11 theme structure-activity 525:542 arg1 studies 557:563 structure-activity relationship studies 525:563 structure-activity relationship studies 525:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 5 12 from hydroxyl 1244:1251 arg1 function 1224:1231 an Fmoc function 1216:1231 an Fmoc function at the C-4 hydroxyl 1216:1251 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 5 13 theme Iduronic 1090:1097 arg1 donors 1104:1109 Iduronic acid donors 1090:1109 Iduronic acid donors 1090:1109 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 4 14 dep it 1028:1029 arg1 group 1051:1055 an ideal protecting group 1031:1055 it an ideal protecting group for HS oligosaccharide assembly 1028:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 3 15 theme levulinoyl 821:830 arg1 ester 838:842 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester 754:842 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester 754:842 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 16 theme Nap 921:923 arg1 ether 926:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 6 17 theme facile 1365:1370 arg1 preparation 1372:1382 the facile preparation 1361:1382 the facile preparation of modular disaccharide building blocks 1361:1422 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 2 18 theme oligosaccharides 504:519 arg1 libraries 488:496 libraries 488:496 libraries of HS oligosaccharides 488:519 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 5 19 theme acid 1099:1102 arg1 donors 1104:1109 Iduronic acid donors 1090:1109 Iduronic acid donors 1090:1109 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 5 20 theme Lev 1195:1197 arg1 ester 1199:1203 a Lev ester 1193:1203 a Lev ester at C-2 1193:1210 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 0 21 theme New 0:2 arg1 acid 15:18 New glucuronic acid 0:18 New glucuronic acid 0:18 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 3 22 theme removable 893:901 arg1 ether 926:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 23 theme 4-acetoxy-2,2-dimethyl 766:787 arg1 butanoyl- 789:797 a permanent 4-acetoxy-2,2-dimethyl butanoyl- 754:797 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) 754:806 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 23 theme 4-acetoxy-2,2-dimethyl 766:787 arg1 PivOAc 800:805 PivOAc 800:805 PivOAc 800:805 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 0 24 theme glucuronic 4:13 arg1 acid 15:18 New glucuronic acid 0:18 New glucuronic acid 0:18 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 3 25 theme C-2 723:725 arg1 hydroxyl 727:734 the C-2 hydroxyl 719:734 the C-2 hydroxyl 719:734 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 0 26 theme heparan 56:62 arg1 oligosaccharides 72:87 heparan sulfate oligosaccharides 56:87 heparan sulfate oligosaccharides 56:87 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 3 27 theme 2-methylnaphthyl 903:918 arg1 ether 926:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 a selectively removable 2-methylnaphthyl (Nap) ether 879:930 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 2 28 theme streamlined 370:380 arg1 approach 382:389 a streamlined approach 368:389 a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies 368:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 2 29 theme blocks 444:449 arg1 preparation 399:409 the preparation 395:409 the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies 395:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 4 30 theme sulfate 1006:1012 arg1 esters 1014:1019 sulfate esters 1006:1019 sulfate esters making it an ideal protecting group for HS oligosaccharide assembly 1006:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 5 31 theme high 1279:1282 arg1 yield 1284:1288 high yield 1279:1288 high yield 1279:1288 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 2 32 theme building 435:442 arg1 blocks 444:449 modular disaccharide building blocks 414:449 modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies 414:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 1 33 theme ligand 266:271 arg1 requirements 273:284 ligand requirements 266:284 ligand requirements for binding and mediating biological activities by these proteins 266:350 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 6 34 theme post-glycosylation 1312:1329 arg1 oxidation 1331:1339 post-glycosylation oxidation 1312:1339 post-glycosylation oxidation 1312:1339 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 2 35 theme modular 414:420 arg1 blocks 444:449 modular disaccharide building blocks 414:449 modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies 414:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 2 36 theme disaccharide 422:433 arg1 blocks 444:449 modular disaccharide building blocks 414:449 modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies 414:563 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 5 37 from C-2 1208:1210 arg1 ester 1199:1203 a Lev ester 1193:1203 a Lev ester at C-2 1193:1210 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 4 38 theme PivOAc 960:965 arg1 ester 967:971 the PivOAc ester 956:971 the PivOAc ester 956:971 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 6 39 theme blocks 1417:1422 arg1 preparation 1372:1382 the facile preparation 1361:1382 the facile preparation of modular disaccharide building blocks 1361:1422 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 3 40 theme acid 611:614 arg1 donors 616:621 glucuronic acid donors 600:621 glucuronic acid donors 600:621 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 41 theme physiological 185:197 arg1 processes 216:224 physiological and pathological processes 185:224 physiological and pathological processes 185:224 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 4 42 theme HS 1061:1062 arg1 assembly 1080:1087 HS oligosaccharide assembly 1061:1087 HS oligosaccharide assembly 1061:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 5 43 theme donating 1143:1150 arg1 properties 1152:1161 more favorable glycosyl donating properties 1119:1161 more favorable glycosyl donating properties 1119:1161 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 6 44 theme building 1408:1415 arg1 blocks 1417:1422 modular disaccharide building blocks 1387:1422 modular disaccharide building blocks 1387:1422 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 3 45 theme butanoyl- 789:797 arg1 ester 838:842 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester 754:842 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester 754:842 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 6 46 theme disaccharide 1395:1406 arg1 blocks 1417:1422 modular disaccharide building blocks 1387:1422 modular disaccharide building blocks 1387:1422 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 3 47 theme permanent 756:764 arg1 butanoyl- 789:797 a permanent 4-acetoxy-2,2-dimethyl butanoyl- 754:797 a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) 754:806 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 47 theme permanent 756:764 arg1 PivOAc 800:805 PivOAc 800:805 PivOAc 800:805 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 48 theme heparan 111:117 arg1 HS 128:129 HS 128:129 HS 128:129 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 1 48 theme heparan 111:117 arg1 sulfate 119:125 heparan sulfate 111:125 heparan sulfate (HS) binding proteins 111:147 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 0 49 theme modular 35:41 arg1 synthesis 43:51 the modular synthesis 31:51 the modular synthesis of heparan sulfate oligosaccharides 31:87 New glucuronic acid donors for the modular synthesis of heparan sulfate oligosaccharides. 24549353 1 50 theme mediating 302:310 arg1 activities 323:332 mediating biological activities 302:332 mediating biological activities 302:332 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 3 51 theme high 681:684 arg1 yields 686:691 high yields 681:691 high yields of coupling products 681:712 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 3 52 theme glucuronic 600:609 arg1 acid 611:614 glucuronic acid 600:614 glucuronic acid donors 600:621 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 53 theme sulfate 119:125 arg1 proteins 140:147 heparan sulfate (HS) binding proteins 111:147 heparan sulfate (HS) binding proteins 111:147 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 3 54 theme C-4 852:854 arg1 hydroxyl 856:863 the C-4 hydroxyl 848:863 the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether 848:930 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 55 theme biological 312:321 arg1 activities 323:332 mediating biological activities 302:332 mediating biological activities 302:332 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 6 56 theme new 1295:1297 arg1 donors 1299:1304 The new donors 1291:1304 The new donors 1291:1304 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 5 57 theme coupling 1258:1265 arg1 products 1267:1274 coupling products 1258:1274 coupling products 1258:1274 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 1 58 theme pathological 203:214 arg1 processes 216:224 physiological and pathological processes 185:224 physiological and pathological processes 185:224 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 1 59 theme processes 216:224 arg1 myriad 175:180 a myriad 173:180 a myriad of physiological and pathological processes 173:224 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 2 60 theme HS 501:502 arg1 oligosaccharides 504:519 HS oligosaccharides 501:519 HS oligosaccharides 501:519 We report here a streamlined approach for the preparation of modular disaccharide building blocks that will facilitate the assembly of libraries of HS oligosaccharides for structure-activity relationship studies. 24549353 5 61 theme glycosyl 1134:1141 arg1 properties 1152:1161 more favorable glycosyl donating properties 1119:1161 more favorable glycosyl donating properties 1119:1161 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 6 62 theme modular 1387:1393 arg1 blocks 1417:1422 modular disaccharide building blocks 1387:1422 modular disaccharide building blocks 1387:1422 The new donors avoid post-glycosylation oxidation and therefore allow the facile preparation of modular disaccharide building blocks. 24549353 4 63 theme oligosaccharide 1064:1078 arg1 assembly 1080:1087 HS oligosaccharide assembly 1061:1087 HS oligosaccharide assembly 1061:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 24549353 5 64 theme Fmoc 1219:1222 arg1 function 1224:1231 an Fmoc function 1216:1231 an Fmoc function at the C-4 hydroxyl 1216:1251 Iduronic acid donors exhibit more favorable glycosyl donating properties and a compound protected with a Lev ester at C-2 and an Fmoc function at the C-4 hydroxyl gave coupling products in high yield. 24549353 3 65 theme products 705:712 arg1 yields 686:691 high yields 681:691 high yields of coupling products 681:712 In particular, we have found that glucuronic acid donors, which usually perform poorly in glycosylations, can give high yields of coupling products when the C-2 hydroxyl is protected with a permanent 4-acetoxy-2,2-dimethyl butanoyl- (PivOAc) or temporary levulinoyl (Lev) ester and the C-4 hydroxyl modified with a selectively removable 2-methylnaphthyl (Nap) ether. 24549353 1 66 theme binding 132:138 arg1 proteins 140:147 heparan sulfate (HS) binding proteins 111:147 heparan sulfate (HS) binding proteins 111:147 Although hundreds of heparan sulfate (HS) binding proteins have been implicated in a myriad of physiological and pathological processes, very little information is known about ligand requirements for binding and mediating biological activities by these proteins. 24549353 4 67 theme ideal 1034:1038 arg1 group 1051:1055 an ideal protecting group 1031:1055 it an ideal protecting group for HS oligosaccharide assembly 1028:1087 It has been shown that the PivOAc ester can be removed without affecting sulfate esters making it an ideal protecting group for HS oligosaccharide assembly. 26515403 9 0 theme sequence 1689:1696 arg1 comparisons 1698:1708 direct amino acid sequence comparisons 1671:1708 direct amino acid sequence comparisons 1671:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 7 1 theme lipid 1226:1230 arg1 modification 1234:1245 lipid A modification 1226:1245 lipid A modification 1226:1245 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 4 2 theme MX 615:616 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 2 3 theme critical 293:300 arg1 This 283:286 This 283:286 This 283:286 This is a critical modification enabling bacteria to resist killing by antimicrobial peptides. 26515403 2 3 theme critical 293:300 arg1 modification 302:313 a critical modification 291:313 a critical modification enabling bacteria to resist killing by antimicrobial peptides 291:375 This is a critical modification enabling bacteria to resist killing by antimicrobial peptides. 26515403 7 4 theme B. 1082:1083 arg1 ArnT 1097:1100 B. cenocepacia ArnT 1082:1100 B. cenocepacia ArnT 1082:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 9 5 theme related 1475:1481 arg1 oligosaccharyltransferases 1483:1508 distantly related oligosaccharyltransferases 1465:1508 distantly related oligosaccharyltransferases 1465:1508 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 6 6 theme Typhimurium 962:972 arg1 domain 990:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 9 7 from families 1529:1536 arg1 oligosaccharyltransferases 1483:1508 distantly related oligosaccharyltransferases 1465:1508 distantly related oligosaccharyltransferases 1465:1508 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 7 8 theme domain 1072:1077 arg1 Deletion 1045:1052 Deletion 1045:1052 Deletion of the C-terminal domain in B. cenocepacia ArnT 1045:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 7 9 with protein 1116:1122 arg1 binding 1160:1166 significantly reduced in vitro binding 1129:1166 significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N 1129:1258 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 8 10 theme in 1264:1265 arg1 model 1295:1299 An in silico predicted structural model 1261:1299 An in silico predicted structural model of ArnT 1261:1307 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 1 11 theme A 247:247 arg1 moiety 249:254 the lipid A moiety 237:254 the lipid A moiety of the lipopolysaccharide 237:280 ArnT is a glycosyltransferase that catalyzes the addition of 4-amino-4-deoxy-l-arabinose (l-Ara4N) to the lipid A moiety of the lipopolysaccharide. 26515403 8 12 theme predicted 1274:1282 arg1 model 1295:1299 An in silico predicted structural model 1261:1299 An in silico predicted structural model of ArnT 1261:1307 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 6 13 theme B 1023:1023 arg1 resistance 1025:1034 polymyxin B resistance 1013:1034 polymyxin B resistance 1013:1034 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 8 14 theme ArnT 1304:1307 arg1 model 1295:1299 An in silico predicted structural model 1261:1299 An in silico predicted structural model of ArnT 1261:1307 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 5 15 from contribution 775:786 arg1 function 810:817 ArnT function 805:817 ArnT function 805:817 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 3 16 theme integral 389:396 arg1 ArnT 378:381 ArnT 378:381 ArnT 378:381 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 3 16 theme integral 389:396 arg1 protein 413:419 an integral inner membrane protein 386:419 an integral inner membrane protein consisting of 13 predicted transmembrane helices 386:468 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 4 17 theme functional 552:561 arg1 motif 563:567 a functional motif 550:567 a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 550:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 5 18 contain have 851:854 arg1 proteins 842:849 these proteins 836:849 these proteins 836:849 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 5 18 contain have 851:854 arg2 mechanism 865:873 a common mechanism 856:873 a common mechanism 856:873 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 8 19 theme tertiary 1332:1339 arg1 structure 1341:1349 the tertiary structure 1328:1349 the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation 1328:1451 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 4 20 theme ArnT 708:711 arg1 proteins 713:720 all ArnT proteins 704:720 all ArnT proteins examined 704:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 0 21 theme bacterial 95:103 arg1 N-oligosaccharyltransferases 105:132 bacterial N-oligosaccharyltransferases 95:132 bacterial N-oligosaccharyltransferases 95:132 ArnT proteins that catalyze the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases. 26515403 6 22 theme C-terminal 979:988 arg1 domain 990:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 3 23 theme large 476:480 arg1 domain 505:510 a large periplasmic C-terminal domain 474:510 a large periplasmic C-terminal domain 474:510 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 0 24 theme ArnT 0:3 arg1 proteins 5:12 ArnT proteins 0:12 ArnT proteins that catalyze the glycosylation of lipopolysaccharide 0:66 ArnT proteins that catalyze the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases. 26515403 9 25 theme structural 1616:1625 arg1 similarity 1627:1636 unexpected structural similarity 1605:1636 unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons 1605:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 3 26 theme membrane 404:411 arg1 ArnT 378:381 ArnT 378:381 ArnT 378:381 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 3 26 theme membrane 404:411 arg1 protein 413:419 an integral inner membrane protein 386:419 an integral inner membrane protein consisting of 13 predicted transmembrane helices 386:468 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 4 27 theme sequence 596:603 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 10 28 theme A 1733:1733 arg1 enzymes 1761:1767 lipid A and protein glycosylation enzymes 1727:1767 lipid A and protein glycosylation enzymes 1727:1767 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 5 29 theme ArnT 805:808 arg1 function 810:817 ArnT function 805:817 ArnT function 805:817 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 9 30 theme lipid 1551:1555 arg1 substrates 1573:1582 lipid and polypeptide substrates 1551:1582 substrates 1573:1582 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 3 31 theme transmembrane 448:460 arg1 helices 462:468 13 predicted transmembrane helices 435:468 13 predicted transmembrane helices 435:468 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 7 32 from ArnT 1097:1100 arg1 Deletion 1045:1052 Deletion 1045:1052 Deletion of the C-terminal domain in B. cenocepacia ArnT 1045:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 8 33 theme bacterial 1381:1389 arg1 oligosaccharyltransferase 1391:1415 a bacterial oligosaccharyltransferase 1379:1415 a bacterial oligosaccharyltransferase involved in protein N-glycosylation 1379:1451 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 8 33 theme bacterial 1381:1389 arg1 PglB 1373:1376 PglB 1373:1376 PglB 1373:1376 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 3 34 theme C-terminal 494:503 arg1 domain 505:510 a large periplasmic C-terminal domain 474:510 a large periplasmic C-terminal domain 474:510 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 7 35 theme in 1151:1152 arg1 binding 1160:1166 significantly reduced in vitro binding 1129:1166 significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N 1129:1258 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 7 36 dep lipid 1173:1177 arg1 unable 1207:1212 unable 1207:1212 unable 1207:1212 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 7 36 dep lipid 1173:1177 arg1 substrate 1193:1201 A fluorescent substrate 1179:1201 A fluorescent substrate 1179:1201 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 4 37 with motif 563:567 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 4 38 theme canonical 576:584 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 8 39 theme protein 1429:1435 arg1 N-glycosylation 1437:1451 protein N-glycosylation 1429:1451 protein N-glycosylation 1429:1451 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 10 40 theme protein 1739:1745 arg1 glycosylation 1747:1759 protein glycosylation 1739:1759 protein glycosylation 1739:1759 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 9 41 theme direct 1671:1676 arg1 comparisons 1698:1708 direct amino acid sequence comparisons 1671:1708 direct amino acid sequence comparisons 1671:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 10 42 theme catalytic 1787:1795 arg1 mechanism 1797:1805 a conserved catalytic mechanism 1775:1805 a conserved catalytic mechanism 1775:1805 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 9 43 theme acid 1684:1687 arg1 comparisons 1698:1708 direct amino acid sequence comparisons 1671:1708 direct amino acid sequence comparisons 1671:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 7 44 from Deletion 1045:1052 arg1 ArnT 1097:1100 B. cenocepacia ArnT 1082:1100 B. cenocepacia ArnT 1082:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 7 45 theme A 1232:1232 arg1 modification 1234:1245 lipid A modification 1226:1245 lipid A modification 1226:1245 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 10 46 theme evolutionary 1821:1832 arg1 divergence 1834:1843 their evolutionary divergence 1815:1843 their evolutionary divergence 1815:1843 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 7 47 from domain 1072:1077 arg1 ArnT 1097:1100 B. cenocepacia ArnT 1082:1100 B. cenocepacia ArnT 1082:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 7 48 theme cenocepacia 1085:1095 arg1 ArnT 1097:1100 B. cenocepacia ArnT 1082:1100 B. cenocepacia ArnT 1082:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 1 49 theme 4-amino-4-deoxy-l-arabinose 196:222 arg1 addition 184:191 the addition 180:191 the addition of 4-amino-4-deoxy-l-arabinose (l-Ara4N) to the lipid A moiety of the lipopolysaccharide 180:280 ArnT is a glycosyltransferase that catalyzes the addition of 4-amino-4-deoxy-l-arabinose (l-Ara4N) to the lipid A moiety of the lipopolysaccharide. 26515403 6 50 theme ArnT 974:977 arg1 domain 990:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 7 51 theme C-terminal 1061:1070 arg1 domain 1072:1077 the C-terminal domain 1057:1077 the C-terminal domain in B. cenocepacia ArnT 1057:1100 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 9 52 theme ArnT 1515:1518 arg1 families 1529:1536 ArnT and PglB families 1515:1536 ArnT and PglB families operating on lipid and polypeptide substrates, respectively 1515:1596 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 9 53 theme PglB 1524:1527 arg1 families 1529:1536 ArnT and PglB families 1515:1536 ArnT and PglB families operating on lipid and polypeptide substrates, respectively 1515:1596 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 6 54 theme cenocepacia 918:928 arg1 domain 990:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain 901:995 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 8 55 theme structural 1284:1293 arg1 model 1295:1299 An in silico predicted structural model 1261:1299 An in silico predicted structural model of ArnT 1261:1307 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 0 56 theme common 74:79 arg1 features 81:88 common features 74:88 common features 74:88 ArnT proteins that catalyze the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases. 26515403 5 57 theme common 858:863 arg1 mechanism 865:873 a common mechanism 856:873 a common mechanism 856:873 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 6 58 theme polymyxin 1013:1021 arg1 resistance 1025:1034 polymyxin B resistance 1013:1034 polymyxin B resistance 1013:1034 We also demonstrate that the Burkholderia cenocepacia and Salmonella enterica serovar Typhimurium ArnT C-terminal domain is required for polymyxin B resistance in vivo. 26515403 4 59 theme first 651:655 arg1 loop 669:672 the first periplasmic loop 647:672 the first periplasmic loop 647:672 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 5 60 theme Site-directed 732:744 arg1 mutagenesis 746:756 Site-directed mutagenesis 732:756 Site-directed mutagenesis 732:756 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 9 61 theme polypeptide 1561:1571 arg1 substrates 1573:1582 lipid and polypeptide substrates 1551:1582 substrates 1573:1582 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 4 62 theme motif 563:567 arg1 identification 532:545 the identification 528:545 the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 528:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 7 63 theme fluorescent 1181:1191 arg1 substrate 1193:1201 A fluorescent substrate 1179:1201 A fluorescent substrate 1179:1201 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 0 64 gly glycosylation 32:44 arg1 lipopolysaccharide 49:66 lipopolysaccharide 49:66 lipopolysaccharide 49:66 ArnT proteins that catalyze the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases. 26515403 8 65 dep in 1264:1265 arg1 silico 1267:1272 silico 1267:1272 silico 1267:1272 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 3 66 theme inner 398:402 arg1 ArnT 378:381 ArnT 378:381 ArnT 378:381 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 3 66 theme inner 398:402 arg1 protein 413:419 an integral inner membrane protein 386:419 an integral inner membrane protein consisting of 13 predicted transmembrane helices 386:468 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 3 67 theme periplasmic 482:492 arg1 domain 505:510 a large periplasmic C-terminal domain 474:510 a large periplasmic C-terminal domain 474:510 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 9 68 theme unexpected 1605:1614 arg1 similarity 1627:1636 unexpected structural similarity 1605:1636 unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons 1605:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 4 69 theme DEXRYAX 605:611 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 7 70 dep in 1151:1152 arg1 vitro 1154:1158 vitro 1154:1158 vitro 1154:1158 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 5 71 theme motif 796:800 arg1 contribution 775:786 the contribution 771:786 the contribution of this motif in ArnT function 771:817 Site-directed mutagenesis demonstrated the contribution of this motif in ArnT function, suggesting that these proteins have a common mechanism. 26515403 8 72 theme PglB 1373:1376 arg1 structure 1341:1349 the tertiary structure 1328:1349 the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation 1328:1451 An in silico predicted structural model of ArnT strongly resembled the tertiary structure of Campylobacter lari PglB, a bacterial oligosaccharyltransferase involved in protein N-glycosylation. 26515403 10 73 theme lipid 1727:1731 arg1 A 1733:1733 lipid A 1727:1733 lipid A 1727:1733 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 7 74 theme reduced 1143:1149 arg1 binding 1160:1166 significantly reduced in vitro binding 1129:1166 significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N 1129:1258 Deletion of the C-terminal domain in B. cenocepacia ArnT resulted in a protein with significantly reduced in vitro binding to a lipid A fluorescent substrate and unable to catalyze lipid A modification with l-Ara4N. 26515403 1 75 theme lipid 241:245 arg1 moiety 249:254 the lipid A moiety 237:254 the lipid A moiety of the lipopolysaccharide 237:280 ArnT is a glycosyltransferase that catalyzes the addition of 4-amino-4-deoxy-l-arabinose (l-Ara4N) to the lipid A moiety of the lipopolysaccharide. 26515403 4 76 theme periplasmic 657:667 arg1 loop 669:672 the first periplasmic loop 647:672 the first periplasmic loop 647:672 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 0 77 theme lipopolysaccharide 49:66 arg1 glycosylation 32:44 the glycosylation 28:44 the glycosylation of lipopolysaccharide 28:66 ArnT proteins that catalyze the glycosylation of lipopolysaccharide share common features with bacterial N-oligosaccharyltransferases. 26515403 4 78 dep YFEKPX 627:632 arg1 W 636:636 W 636:636 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 4 78 dep YFEKPX 627:632 arg1 4 634:634 4 634:634 4 634:634 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 3 79 theme predicted 438:446 arg1 helices 462:468 13 predicted transmembrane helices 435:468 13 predicted transmembrane helices 435:468 ArnT is an integral inner membrane protein consisting of 13 predicted transmembrane helices and a large periplasmic C-terminal domain. 26515403 4 80 theme consensus 586:594 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 10 81 theme glycosylation 1747:1759 arg1 enzymes 1761:1767 lipid A and protein glycosylation enzymes 1727:1767 lipid A and protein glycosylation enzymes 1727:1767 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 4 82 theme GXWX 620:623 arg1 YFEKPX 627:632 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined 574:729 We report here the identification of a functional motif with a canonical consensus sequence DEXRYAX(5)MX(3)GXWX(9)YFEKPX(4)W spanning the first periplasmic loop, which is highly conserved in all ArnT proteins examined. 26515403 2 83 theme antimicrobial 354:366 arg1 peptides 368:375 antimicrobial peptides 354:375 antimicrobial peptides 354:375 This is a critical modification enabling bacteria to resist killing by antimicrobial peptides. 26515403 9 84 theme amino 1678:1682 arg1 comparisons 1698:1708 direct amino acid sequence comparisons 1671:1708 direct amino acid sequence comparisons 1671:1708 Therefore, distantly related oligosaccharyltransferases from ArnT and PglB families operating on lipid and polypeptide substrates, respectively, share unexpected structural similarity that could not be predicted from direct amino acid sequence comparisons. 26515403 10 85 theme conserved 1777:1785 arg1 mechanism 1797:1805 a conserved catalytic mechanism 1775:1805 a conserved catalytic mechanism 1775:1805 We propose that lipid A and protein glycosylation enzymes share a conserved catalytic mechanism despite their evolutionary divergence. 26515403 1 86 theme lipopolysaccharide 263:280 arg1 moiety 249:254 the lipid A moiety 237:254 the lipid A moiety of the lipopolysaccharide 237:280 ArnT is a glycosyltransferase that catalyzes the addition of 4-amino-4-deoxy-l-arabinose (l-Ara4N) to the lipid A moiety of the lipopolysaccharide. 28729420 7 0 theme lower 1535:1539 arg1 efficiency 1541:1550 much lower efficiency 1530:1550 much lower efficiency than the Man5GlcNAc2 peptide 1530:1579 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 6 1 theme Man5GlcNAc2 1120:1130 arg1 glycan 1132:1137 Man5GlcNAc2 glycan 1120:1137 Man5GlcNAc2 glycan 1120:1137 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 8 2 theme GlcNAc 1700:1705 arg1 moiety 1707:1712 the α1,3-arm GlcNAc moiety 1687:1712 the α1,3-arm GlcNAc moiety 1687:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 0 3 gly fucosylation 105:116 arg1 N-glycans 121:129 N-glycans 121:129 N-glycans lacking α1,3-arm GlcNAc 121:153 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 3 4 from expression 531:540 arg1 line 606:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 5 gly core-fucosylated 646:661 arg1 Man5GlcNAc2 704:714 the oligomannose substrate Man5GlcNAc2 677:714 the oligomannose substrate Man5GlcNAc2 677:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 5 gly core-fucosylated 646:661 arg1 glycoforms 663:672 fully core-fucosylated glycoforms 640:672 fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2 640:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 6 theme core 751:754 arg1 fucosylation 756:767 core fucosylation 751:767 core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells 751:816 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 1 7 gly fucosylation 220:231 arg1 N-glycans 236:244 N-glycans 236:244 N-glycans 236:244 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 1 7 gly fucosylation 220:231 arg1 biosynthesis 253:264 the biosynthesis 249:264 the biosynthesis of glycoproteins 249:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 2 8 theme in 306:307 arg1 studies 315:321 intensive in vitro studies 296:321 intensive in vitro studies with crude extract or purified enzyme 296:359 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 4 9 theme in 889:890 arg1 activity 898:905 its in vitro activity 885:905 its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins 885:977 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 2 10 from attachment 380:389 arg1 N-glycan 430:437 N-glycan 430:437 N-glycan 430:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 10 from attachment 380:389 arg1 arm 423:425 the α1,3 mannose arm 406:425 the α1,3 mannose arm of N-glycan 406:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 6 11 theme appropriate 1191:1201 arg1 context 1219:1225 an appropriate protein/peptide context 1188:1225 an appropriate protein/peptide context 1188:1225 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 6 12 attach derived 1286:1292 arg2 polypeptide 1274:1284 a V3 polypeptide 1269:1284 a V3 polypeptide derived from HIV-1 gp120 1269:1309 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 6 12 attach derived 1286:1292 arg1 gp120 1305:1309 HIV-1 gp120 1299:1309 HIV-1 gp120 1299:1309 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 3 13 theme α1,3-arm 793:800 arg1 GlcNAc 802:807 an α1,3-arm GlcNAc 790:807 an α1,3-arm GlcNAc in cells 790:816 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 14 theme cell 601:604 arg1 line 606:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 1 15 theme core 215:218 arg1 fucosylation 220:231 the core fucosylation 211:231 the core fucosylation of N-glycans in the biosynthesis of glycoproteins 211:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 4 16 theme glycopeptides 946:958 arg1 array 917:921 an array 914:921 an array of selected N-glycans, glycopeptides, and glycoproteins 914:977 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 0 17 theme α1,3-arm 139:146 arg1 GlcNAc 148:153 α1,3-arm GlcNAc 139:153 α1,3-arm GlcNAc 139:153 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 5 18 theme FUT8 1094:1097 arg1 substrates 1099:1108 FUT8 substrates 1094:1108 FUT8 substrates 1094:1108 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 5 18 theme FUT8 1094:1097 arg1 N-glycans 1033:1041 free N-glycans 1028:1041 free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety 1028:1084 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 2 19 theme crude 328:332 arg1 extract 334:340 crude extract 328:340 crude extract 328:340 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 6 20 theme V3 1271:1272 arg1 polypeptide 1274:1284 a V3 polypeptide 1269:1284 a V3 polypeptide derived from HIV-1 gp120 1269:1309 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 8 21 dep in 1624:1625 arg1 vitro 1627:1631 vitro 1627:1631 vitro 1627:1631 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 9 22 theme adequate 1784:1791 arg1 elements 1804:1811 other adequate structural elements 1778:1811 other adequate structural elements 1778:1811 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 7 23 theme V3 1412:1413 arg1 context 1427:1433 the V3 polypeptide context 1408:1433 the V3 polypeptide context 1408:1433 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 6 24 theme chloroformate-protected 1342:1364 arg1 moiety 1370:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 5 25 theme GlcNAc 1072:1077 arg1 moiety 1079:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 1 26 from fucosylation 220:231 arg1 biosynthesis 253:264 the biosynthesis 249:264 the biosynthesis of glycoproteins 249:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 4 27 gly glycopeptides 946:958 arg2 glycopeptides 946:958 glycopeptides 946:958 glycopeptides 946:958 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 4 28 theme N-glycans 935:943 arg1 array 917:921 an array 914:921 an array of selected N-glycans, glycopeptides, and glycoproteins 914:977 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 6 29 theme erythropoietin 1245:1258 arg1 protein 1260:1266 the erythropoietin protein 1241:1266 the erythropoietin protein 1241:1266 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 6 30 theme simple 1317:1322 arg1 moiety 1370:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 3 31 theme core-fucosylated 646:661 arg1 glycoforms 663:672 fully core-fucosylated glycoforms 640:672 fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2 640:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 2 32 with studies 315:321 arg1 extract 334:340 crude extract 328:340 crude extract 328:340 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 32 with studies 315:321 arg1 enzyme 354:359 purified enzyme 345:359 purified enzyme 345:359 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 1 33 gly glycoproteins 269:281 arg1 glycoproteins 269:281 glycoproteins 269:281 glycoproteins 269:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 9 34 from context 1767:1773 arg1 substrate 1829:1837 the acceptor substrate 1816:1837 the acceptor substrate 1816:1837 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 9 35 theme core 1860:1863 arg1 fucosylation 1865:1876 the core fucosylation 1856:1876 the core fucosylation 1856:1876 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 6 36 gly core-fucosylated 1160:1175 arg1 glycan 1132:1137 Man5GlcNAc2 glycan 1120:1137 Man5GlcNAc2 glycan 1120:1137 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 2 37 theme mannose 415:421 arg1 N-glycan 430:437 N-glycan 430:437 N-glycan 430:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 37 theme mannose 415:421 arg1 arm 423:425 the α1,3 mannose arm 406:425 the α1,3 mannose arm of N-glycan 406:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 0 38 theme N-glycans 121:129 arg1 fucosylation 105:116 core fucosylation 100:116 core fucosylation of N-glycans lacking α1,3-arm GlcNAc 100:153 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 2 39 theme core 471:474 arg1 fucosylation 476:487 FUT8-catalyzed core fucosylation 456:487 FUT8-catalyzed core fucosylation 456:487 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 0 40 theme substrate 15:23 arg1 specificity 25:35 the substrate specificity 11:35 the substrate specificity of mammalian α1,6-fucosyltransferase 11:72 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 3 41 dep knock-out 570:578 arg1 FUT8-overexpressing 581:599 FUT8-overexpressing 581:599 FUT8-overexpressing 581:599 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 8 42 theme first 1608:1612 arg1 report 1614:1619 the first report 1604:1619 the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1604:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 3 43 theme oligomannose 681:692 arg1 Man5GlcNAc2 704:714 the oligomannose substrate Man5GlcNAc2 677:714 the oligomannose substrate Man5GlcNAc2 677:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 8 44 gly fucosylation 1653:1664 arg1 N-glycans 1669:1677 N-glycans 1669:1677 N-glycans lacking the α1,3-arm GlcNAc moiety 1669:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 4 45 theme FUT8 867:870 arg1 specificity 852:862 the substrate specificity 838:862 the substrate specificity of FUT8 838:870 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 0 46 theme α1,6-fucosyltransferase 50:72 arg1 specificity 25:35 the substrate specificity 11:35 the substrate specificity of mammalian α1,6-fucosyltransferase 11:72 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 5 47 theme α1,3-arm 1063:1070 arg1 moiety 1079:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 9 48 theme structural 1793:1802 arg1 elements 1804:1811 other adequate structural elements 1778:1811 other adequate structural elements 1778:1811 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 6 49 from core-fucosylated 1160:1175 arg1 context 1219:1225 an appropriate protein/peptide context 1188:1225 an appropriate protein/peptide context 1188:1225 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 8 50 theme core 1648:1651 arg1 fucosylation 1653:1664 in vitro FUT8-catalyzed core fucosylation 1624:1664 in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1624:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 3 51 theme Man5GlcNAc2 704:714 arg1 glycoforms 663:672 fully core-fucosylated glycoforms 640:672 fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2 640:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 2 52 theme GlcNAc 396:401 arg1 essential 442:450 essential 442:450 essential 442:450 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 52 theme GlcNAc 396:401 arg1 attachment 380:389 the attachment 376:389 the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan 376:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 53 attach attachment 380:389 arg2 GlcNAc 396:401 a GlcNAc 394:401 a GlcNAc 394:401 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 53 attach attachment 380:389 arg3 arm 423:425 the α1,3 mannose arm 406:425 the α1,3 mannose arm of N-glycan 406:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 53 attach attachment 380:389 arg3 N-glycan 430:437 N-glycan 430:437 N-glycan 430:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 7 54 theme bi-antennary 1445:1456 arg1 core-fucosylated 1494:1509 core-fucosylated 1494:1509 core-fucosylated 1494:1509 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 54 theme bi-antennary 1445:1456 arg1 N-glycan 1471:1478 a mature bi-antennary complex-type N-glycan 1436:1478 a mature bi-antennary complex-type N-glycan 1436:1478 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 8 55 theme α1,3-arm 1691:1698 arg1 moiety 1707:1712 the α1,3-arm GlcNAc moiety 1687:1712 the α1,3-arm GlcNAc moiety 1687:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 3 56 gly glycoforms 663:672 arg1 Man5GlcNAc2 704:714 the oligomannose substrate Man5GlcNAc2 677:714 the oligomannose substrate Man5GlcNAc2 677:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 57 theme N-glycans 772:780 arg1 fucosylation 756:767 core fucosylation 751:767 core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells 751:816 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 6 58 theme HIV-1 1299:1303 arg1 gp120 1305:1309 HIV-1 gp120 1299:1309 HIV-1 gp120 1299:1309 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 3 59 theme erythropoietin 545:558 arg1 expression 531:540 expression 531:540 expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line 531:609 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 6 60 theme protein/peptide 1203:1217 arg1 context 1219:1225 an appropriate protein/peptide context 1188:1225 an appropriate protein/peptide context 1188:1225 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 5 61 with Consistent 980:989 arg1 studies 1005:1011 previous studies 996:1011 previous studies 996:1011 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 2 62 theme purified 345:352 arg1 enzyme 354:359 purified enzyme 345:359 purified enzyme 345:359 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 9 63 theme appropriate 1743:1753 arg1 context 1767:1773 an appropriate polypeptide context 1740:1773 an appropriate polypeptide context 1740:1773 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 7 64 theme complex-type 1458:1469 arg1 core-fucosylated 1494:1509 core-fucosylated 1494:1509 core-fucosylated 1494:1509 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 64 theme complex-type 1458:1469 arg1 N-glycan 1471:1478 a mature bi-antennary complex-type N-glycan 1436:1478 a mature bi-antennary complex-type N-glycan 1436:1478 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 3 65 theme knock-out 570:578 arg1 line 606:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 a GnTI knock-out, FUT8-overexpressing cell line 563:609 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 7 66 theme mature 1438:1443 arg1 core-fucosylated 1494:1509 core-fucosylated 1494:1509 core-fucosylated 1494:1509 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 66 theme mature 1438:1443 arg1 N-glycan 1471:1478 a mature bi-antennary complex-type N-glycan 1436:1478 a mature bi-antennary complex-type N-glycan 1436:1478 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 1 67 theme N-glycans 236:244 arg1 fucosylation 220:231 the core fucosylation 211:231 the core fucosylation of N-glycans in the biosynthesis of glycoproteins 211:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 9 68 theme other 1778:1782 arg1 elements 1804:1811 other adequate structural elements 1778:1811 other adequate structural elements 1778:1811 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 6 69 theme Asn 1366:1368 arg1 moiety 1370:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 9 70 theme polypeptide 1755:1765 arg1 context 1767:1773 an appropriate polypeptide context 1740:1773 an appropriate polypeptide context 1740:1773 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 7 71 theme polypeptide 1415:1425 arg1 context 1427:1433 the V3 polypeptide context 1408:1433 the V3 polypeptide context 1408:1433 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 6 72 theme 9-fluorenylmethyl 1324:1340 arg1 moiety 1370:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 a simple 9-fluorenylmethyl chloroformate-protected Asn moiety 1315:1375 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 2 73 theme intensive 296:304 arg1 studies 315:321 intensive in vitro studies 296:321 intensive in vitro studies with crude extract or purified enzyme 296:359 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 9 74 from elements 1804:1811 arg1 substrate 1829:1837 the acceptor substrate 1816:1837 the acceptor substrate 1816:1837 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 4 75 theme selected 926:933 arg1 N-glycans 935:943 selected N-glycans 926:943 selected N-glycans 926:943 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 1 76 theme mammalian 160:168 arg1 α1,6-fucosyltransferase 170:192 The mammalian α1,6-fucosyltransferase 156:192 The mammalian α1,6-fucosyltransferase (FUT8) 156:199 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 1 76 theme mammalian 160:168 arg1 FUT8 195:198 FUT8 195:198 FUT8 195:198 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 28729420 9 77 theme acceptor 1820:1827 arg1 substrate 1829:1837 the acceptor substrate 1816:1837 the acceptor substrate 1816:1837 Our results suggest that an appropriate polypeptide context or other adequate structural elements in the acceptor substrate could facilitate the core fucosylation by FUT8. 28729420 2 78 dep in 306:307 arg1 vitro 309:313 vitro 309:313 vitro 309:313 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 0 79 theme core 100:103 arg1 fucosylation 105:116 core fucosylation 100:116 core fucosylation of N-glycans lacking α1,3-arm GlcNAc 100:153 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 5 80 theme previous 996:1003 arg1 studies 1005:1011 previous studies 996:1011 previous studies 996:1011 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 4 81 theme glycoproteins 965:977 arg1 array 917:921 an array 914:921 an array of selected N-glycans, glycopeptides, and glycoproteins 914:977 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 6 82 from context 1219:1225 arg1 core-fucosylated 1160:1175 core-fucosylated 1160:1175 core-fucosylated 1160:1175 However, Man5GlcNAc2 glycan could be efficiently core-fucosylated by FUT8 in an appropriate protein/peptide context, such as with the erythropoietin protein, a V3 polypeptide derived from HIV-1 gp120, or a simple 9-fluorenylmethyl chloroformate-protected Asn moiety. 28729420 2 83 theme α1,3 410:413 arg1 N-glycan 430:437 N-glycan 430:437 N-glycan 430:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 83 theme α1,3 410:413 arg1 arm 423:425 the α1,3 mannose arm 406:425 the α1,3 mannose arm of N-glycan 406:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 4 84 dep in 889:890 arg1 vitro 892:896 vitro 892:896 vitro 892:896 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 4 85 gly glycoproteins 965:977 arg1 glycoproteins 965:977 glycoproteins 965:977 glycoproteins 965:977 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 7 86 gly core-fucosylated 1494:1509 arg2 efficiency 1541:1550 much lower efficiency 1530:1550 much lower efficiency than the Man5GlcNAc2 peptide 1530:1579 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 86 gly core-fucosylated 1494:1509 arg1 core-fucosylated 1494:1509 core-fucosylated 1494:1509 core-fucosylated 1494:1509 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 86 gly core-fucosylated 1494:1509 arg1 N-glycan 1471:1478 a mature bi-antennary complex-type N-glycan 1436:1478 a mature bi-antennary complex-type N-glycan 1436:1478 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 2 87 theme FUT8-catalyzed 456:469 arg1 fucosylation 476:487 FUT8-catalyzed core fucosylation 456:487 FUT8-catalyzed core fucosylation 456:487 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 3 88 from GlcNAc 802:807 arg1 cells 812:816 cells 812:816 cells 812:816 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 3 89 gly fucosylation 756:767 arg1 N-glycans 772:780 N-glycans 772:780 N-glycans lacking an α1,3-arm GlcNAc in cells 772:816 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 2 90 theme N-glycan 430:437 arg1 N-glycan 430:437 N-glycan 430:437 N-glycan 430:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 2 90 theme N-glycan 430:437 arg1 arm 423:425 the α1,3 mannose arm 406:425 the α1,3 mannose arm of N-glycan 406:437 Previously, intensive in vitro studies with crude extract or purified enzyme concluded that the attachment of a GlcNAc on the α1,3 mannose arm of N-glycan is essential for FUT8-catalyzed core fucosylation. 28729420 0 91 theme mammalian 40:48 arg1 α1,6-fucosyltransferase 50:72 mammalian α1,6-fucosyltransferase 40:72 mammalian α1,6-fucosyltransferase 40:72 Revisiting the substrate specificity of mammalian α1,6-fucosyltransferase reveals that it catalyzes core fucosylation of N-glycans lacking α1,3-arm GlcNAc. 28729420 8 92 theme in 1624:1625 arg1 fucosylation 1653:1664 in vitro FUT8-catalyzed core fucosylation 1624:1664 in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1624:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 3 93 theme glycoforms 663:672 arg1 production 626:635 the production 622:635 the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2 622:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 4 94 theme substrate 842:850 arg1 specificity 852:862 the substrate specificity 838:862 the substrate specificity of FUT8 838:870 Here, we revisited the substrate specificity of FUT8 by examining its in vitro activity toward an array of selected N-glycans, glycopeptides, and glycoproteins. 28729420 5 95 theme unmasked 1054:1061 arg1 moiety 1079:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 an unmasked α1,3-arm GlcNAc moiety 1051:1084 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 8 96 theme FUT8-catalyzed 1633:1646 arg1 fucosylation 1653:1664 in vitro FUT8-catalyzed core fucosylation 1624:1664 in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1624:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 7 97 theme Man5GlcNAc2 1561:1571 arg1 peptide 1573:1579 the Man5GlcNAc2 peptide 1557:1579 the Man5GlcNAc2 peptide 1557:1579 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 8 98 theme fucosylation 1653:1664 arg1 report 1614:1619 the first report 1604:1619 the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1604:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 3 99 theme substrate 694:702 arg1 Man5GlcNAc2 704:714 the oligomannose substrate Man5GlcNAc2 677:714 the oligomannose substrate Man5GlcNAc2 677:714 In contrast, we have recently shown that expression of erythropoietin in a GnTI knock-out, FUT8-overexpressing cell line results in the production of fully core-fucosylated glycoforms of the oligomannose substrate Man5GlcNAc2, suggesting that FUT8 can catalyze core fucosylation of N-glycans lacking an α1,3-arm GlcNAc in cells. 28729420 7 100 from efficiency 1541:1550 arg1 core-fucosylated 1494:1509 core-fucosylated 1494:1509 core-fucosylated 1494:1509 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 7 100 from efficiency 1541:1550 arg1 N-glycan 1471:1478 a mature bi-antennary complex-type N-glycan 1436:1478 a mature bi-antennary complex-type N-glycan 1436:1478 Interestingly, when placed in the V3 polypeptide context, a mature bi-antennary complex-type N-glycan also could be core-fucosylated by FUT8, albeit at much lower efficiency than the Man5GlcNAc2 peptide. 28729420 5 101 theme free 1028:1031 arg1 substrates 1099:1108 FUT8 substrates 1094:1108 FUT8 substrates 1094:1108 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 5 101 theme free 1028:1031 arg1 N-glycans 1033:1041 free N-glycans 1028:1041 free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety 1028:1084 Consistent with previous studies, we found that free N-glycans lacking an unmasked α1,3-arm GlcNAc moiety are not FUT8 substrates. 28729420 8 102 theme N-glycans 1669:1677 arg1 fucosylation 1653:1664 in vitro FUT8-catalyzed core fucosylation 1624:1664 in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety 1624:1712 This study represents the first report of in vitro FUT8-catalyzed core fucosylation of N-glycans lacking the α1,3-arm GlcNAc moiety. 28729420 1 103 theme glycoproteins 269:281 arg1 biosynthesis 253:264 the biosynthesis 249:264 the biosynthesis of glycoproteins 249:281 The mammalian α1,6-fucosyltransferase (FUT8) catalyzes the core fucosylation of N-glycans in the biosynthesis of glycoproteins. 25187161 3 0 theme -/- 672:674 arg1 TCRδ 667:670 TCRδ(-/-) and WT mice 667:687 TCRδ 667:670 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 6 1 from antennae 1025:1032 arg1 colon 1047:1051 TCRδ(-/-) colon 1037:1051 TCRδ(-/-) colon 1037:1051 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 6 1 from antennae 1025:1032 arg1 intestine 1063:1071 small intestine 1057:1071 small intestine 1057:1071 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 2 2 theme in-depth 382:389 arg1 comparison 410:419 an in-depth mass spectrometric comparison 379:419 an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 379:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 6 3 theme WT 1123:1124 arg1 mice 1126:1129 WT mice 1123:1129 WT mice 1123:1129 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 3 4 theme mice 684:687 arg1 intestine 644:652 small intestine 638:652 small intestine 638:652 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 3 4 theme mice 684:687 arg1 colon 658:662 colon 658:662 colon 658:662 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 7 5 dep intestine 1298:1306 arg1 -/- 1288:1290 -/- 1288:1290 -/- 1288:1290 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 6 6 theme structural 1088:1097 arg1 diversity 1099:1107 altered structural diversity 1080:1107 altered structural diversity 1080:1107 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 3 7 theme nonsulfated 554:564 arg1 types 596:600 69 nonsulfated and 59 sulfated compositional types 551:600 69 nonsulfated and 59 sulfated compositional types of O-glycans 551:613 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 5 8 theme TCRδ 814:817 arg1 colons 824:829 TCRδ(-/-) colons 814:829 TCRδ(-/-) colons 814:829 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 7 9 theme small 1292:1296 arg1 intestine 1298:1306 the TCRδ(-/-)small intestine 1279:1306 the TCRδ(-/-)small intestine 1279:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 7 10 from differences 1155:1165 arg1 species 1185:1191 the sialylated species 1170:1191 the sialylated species 1170:1191 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 5 11 theme core-3 995:1000 arg1 structures 1002:1011 core-3 structures 995:1011 core-3 structures 995:1011 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 3 12 theme compositional 582:594 arg1 types 596:600 69 nonsulfated and 59 sulfated compositional types 551:600 69 nonsulfated and 59 sulfated compositional types of O-glycans 551:613 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 8 13 theme novel 1389:1393 arg1 insights 1405:1412 novel molecular insights 1389:1412 novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis 1389:1476 These findings provide novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis. 25187161 7 14 theme TCRδ 1283:1286 arg1 intestine 1298:1306 the TCRδ(-/-)small intestine 1279:1306 the TCRδ(-/-)small intestine 1279:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 7 15 dep TCRδ 1205:1208 arg1 -/- 1210:1212 -/- 1210:1212 -/- 1210:1212 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 5 16 theme increased 888:896 arg1 proportion 898:907 an increased proportion 885:907 an increased proportion of core-1 structures 885:928 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 2 17 theme wild-type 512:520 arg1 littermates 527:537 their wild-type (WT) littermates 506:537 their wild-type (WT) littermates 506:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 4 18 gly glycosylation 752:764 arg1 mice 779:782 TCRδ(-/-) mice 769:782 TCRδ(-/-) mice 769:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 1 19 theme receptor-bearing 165:180 arg1 IELs 214:217 γδ IELs 211:217 γδ IELs 211:217 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 1 19 theme receptor-bearing 165:180 arg1 lymphocytes 198:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes 144:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) 144:218 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 0 20 theme intestinal 79:88 arg1 profile 99:105 an altered intestinal O-glycan profile 68:105 an altered intestinal O-glycan profile 68:105 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 5 21 theme core-2 863:868 arg1 structures 870:879 core-2 structures 863:879 core-2 structures 863:879 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 2 22 theme littermates 527:537 arg1 profile 455:461 the intestinal O-glycosylation profile 424:461 the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 424:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 5 23 contain contained 831:839 arg1 colons 824:829 TCRδ(-/-) colons 814:829 TCRδ(-/-) colons 814:829 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 5 23 contain contained 831:839 arg2 proportion 849:858 a lower proportion 841:858 a lower proportion of core-2 structures 841:879 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 5 23 contain contained 831:839 arg2 proportion 898:907 an increased proportion 885:907 an increased proportion of core-1 structures 885:928 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 4 24 from glycosylation 752:764 arg1 mice 779:782 TCRδ(-/-) mice 769:782 TCRδ(-/-) mice 769:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 5 25 theme TCRδ 938:941 arg1 intestines 954:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 1 26 theme γδ 155:156 arg1 IELs 214:217 γδ IELs 211:217 γδ IELs 211:217 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 1 26 theme γδ 155:156 arg1 lymphocytes 198:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes 144:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) 144:218 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 0 27 from lymphocytes 48:58 arg1 deficient 5:13 deficient 5:13 deficient 5:13 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 4 28 theme TCRδ 769:772 arg1 mice 779:782 TCRδ(-/-) mice 769:782 TCRδ(-/-) mice 769:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 2 29 theme O-glycosylation 439:453 arg1 profile 455:461 the intestinal O-glycosylation profile 424:461 the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 424:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 5 30 theme -/- 943:945 arg1 intestines 954:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 0 31 theme altered 71:77 arg1 profile 99:105 an altered intestinal O-glycan profile 68:105 an altered intestinal O-glycan profile 68:105 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 4 32 theme structural 715:724 arg1 differences 726:736 structural differences 715:736 structural differences in intestinal glycosylation in TCRδ(-/-) mice 715:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 6 33 theme glycan 1018:1023 arg1 antennae 1025:1032 The glycan antennae 1014:1032 The glycan antennae in TCRδ(-/-) colon and small intestine 1014:1071 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 6 34 dep TCRδ 1037:1040 arg1 -/- 1042:1044 -/- 1042:1044 -/- 1042:1044 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 5 35 theme structures 919:928 arg1 proportion 898:907 an increased proportion 885:907 an increased proportion of core-1 structures 885:928 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 5 35 theme structures 919:928 arg1 proportion 849:858 a lower proportion 841:858 a lower proportion of core-2 structures 841:879 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 7 36 theme sialylated 1236:1245 arg1 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 0 37 theme γδ 29:30 arg1 lymphocytes 48:58 intestinal γδ intraepithelial lymphocytes 18:58 intestinal γδ intraepithelial lymphocytes 18:58 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 5 38 theme decreased 971:979 arg1 percentage 981:990 a decreased percentage 969:990 a decreased percentage of core-3 structures 969:1011 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 3 39 theme small 638:642 arg1 intestine 644:652 small intestine 638:652 small intestine 638:652 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 7 40 theme WT 1219:1220 arg1 mice 1222:1225 the TCRδ(-/-) and WT mice 1201:1225 mice 1222:1225 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 4 41 theme -/- 774:776 arg1 mice 779:782 TCRδ(-/-) mice 769:782 TCRδ(-/-) mice 769:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 5 42 theme small 948:952 arg1 intestines 954:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 3 43 dep intestine 644:652 arg1 the 634:636 the 634:636 the 634:636 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 5 44 contain had 965:967 arg1 intestines 954:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) small intestines 938:963 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 5 44 contain had 965:967 arg2 percentage 981:990 a decreased percentage 969:990 a decreased percentage of core-3 structures 969:1011 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 3 45 theme WT 681:682 arg1 mice 684:687 TCRδ(-/-) and WT mice 667:687 mice 684:687 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 2 46 theme mass 391:394 arg1 comparison 410:419 an in-depth mass spectrometric comparison 379:419 an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 379:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 7 47 with TCRδ 1205:1208 arg1 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 8 48 theme γδ 1439:1440 arg1 IELs 1442:1445 γδ IELs 1439:1445 γδ IELs 1439:1445 These findings provide novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis. 25187161 7 49 theme sulfation 1321:1329 arg1 pattern 1331:1337 the sulfation pattern 1317:1337 the sulfation pattern 1317:1337 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 7 50 with mice 1222:1225 arg1 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 6 51 theme altered 1080:1086 arg1 diversity 1099:1107 altered structural diversity 1080:1107 altered structural diversity 1080:1107 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 1 52 theme T-cell 158:163 arg1 IELs 214:217 γδ IELs 211:217 γδ IELs 211:217 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 1 52 theme T-cell 158:163 arg1 lymphocytes 198:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes 144:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) 144:218 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 3 53 theme types 596:600 arg1 total 542:546 A total 540:546 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans 540:613 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 1 54 theme multifaceted 227:238 arg1 role 240:243 a multifaceted role 225:243 a multifaceted role 225:243 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 5 55 theme structures 1002:1011 arg1 percentage 981:990 a decreased percentage 969:990 a decreased percentage of core-3 structures 969:1011 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 3 56 theme sulfated 573:580 arg1 types 596:600 69 nonsulfated and 59 sulfated compositional types 551:600 69 nonsulfated and 59 sulfated compositional types of O-glycans 551:613 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 5 57 theme structures 870:879 arg1 proportion 898:907 an increased proportion 885:907 an increased proportion of core-1 structures 885:928 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 5 57 theme structures 870:879 arg1 proportion 849:858 a lower proportion 841:858 a lower proportion of core-2 structures 841:879 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 8 58 theme molecular 1395:1403 arg1 insights 1405:1412 novel molecular insights 1389:1412 novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis 1389:1476 These findings provide novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis. 25187161 7 59 located found 1258:1262 arg2 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 7 59 located found 1258:1262 arg1 intestine 1298:1306 the TCRδ(-/-)small intestine 1279:1306 the TCRδ(-/-)small intestine 1279:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 1 60 theme Intestinal 144:153 arg1 IELs 214:217 γδ IELs 211:217 γδ IELs 211:217 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 1 60 theme Intestinal 144:153 arg1 lymphocytes 198:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes 144:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) 144:218 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 7 61 theme sialylated 1174:1183 arg1 species 1185:1191 the sialylated species 1170:1191 the sialylated species 1170:1191 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 3 62 theme O-glycans 605:613 arg1 types 596:600 69 nonsulfated and 59 sulfated compositional types 551:600 69 nonsulfated and 59 sulfated compositional types of O-glycans 551:613 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 5 63 theme lower 843:847 arg1 proportion 849:858 a lower proportion 841:858 a lower proportion of core-2 structures 841:879 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 0 64 theme O-glycan 90:97 arg1 profile 99:105 an altered intestinal O-glycan profile 68:105 an altered intestinal O-glycan profile 68:105 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 2 65 contain carried 367:373 arg2 comparison 410:419 an in-depth mass spectrometric comparison 379:419 an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 379:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 2 65 contain carried 367:373 arg1 we 364:365 we 364:365 we 364:365 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 7 66 gly sialylated 1236:1245 arg1 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 1 67 theme intraepithelial 182:196 arg1 IELs 214:217 γδ IELs 211:217 γδ IELs 211:217 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 1 67 theme intraepithelial 182:196 arg1 lymphocytes 198:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes 144:208 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) 144:218 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 0 68 from deficient 5:13 arg1 lymphocytes 48:58 intestinal γδ intraepithelial lymphocytes 18:58 intestinal γδ intraepithelial lymphocytes 18:58 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 7 69 theme significant 1143:1153 arg1 differences 1155:1165 significant differences 1143:1165 significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1143:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 4 70 theme intestinal 741:750 arg1 glycosylation 752:764 intestinal glycosylation 741:764 intestinal glycosylation in TCRδ(-/-) mice 741:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 0 71 theme wild-type 121:129 arg1 littermates 131:141 wild-type littermates 121:141 wild-type littermates 121:141 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 6 72 theme TCRδ 1037:1040 arg1 colon 1047:1051 TCRδ(-/-) colon 1037:1051 TCRδ(-/-) colon 1037:1051 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 8 73 theme gut 1462:1464 arg1 homeostasis 1466:1476 gut homeostasis 1462:1476 gut homeostasis 1462:1476 These findings provide novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis. 25187161 2 74 gly O-glycosylation 439:453 arg1 mice 466:469 mice 466:469 mice lacking γδ IELs (TCRδ(-/-)) 466:497 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 2 74 gly O-glycosylation 439:453 arg1 littermates 527:537 their wild-type (WT) littermates 506:537 their wild-type (WT) littermates 506:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 2 75 theme intestinal 428:437 arg1 profile 455:461 the intestinal O-glycosylation profile 424:461 the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 424:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 0 76 theme deficient 5:13 arg1 Mice 0:3 Mice 0:3 Mice deficient in intestinal γδ intraepithelial lymphocytes 0:58 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 2 77 dep TCRδ 488:491 arg1 -/- 493:495 -/- 493:495 -/- 493:495 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 4 78 from differences 726:736 arg1 glycosylation 752:764 intestinal glycosylation 741:764 intestinal glycosylation in TCRδ(-/-) mice 741:782 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 8 79 theme IELs 1442:1445 arg1 role 1431:1434 the role 1427:1434 the role of γδ IELs in maintaining gut homeostasis 1427:1476 These findings provide novel molecular insights underpinning the role of γδ IELs in maintaining gut homeostasis. 25187161 7 80 theme Tn 1247:1248 arg1 antigen 1250:1256 the sialylated Tn antigen 1232:1256 the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine 1232:1306 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 7 81 gly sialylated 1174:1183 arg1 species 1185:1191 the sialylated species 1170:1191 the sialylated species 1170:1191 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 0 82 theme intestinal 18:27 arg1 lymphocytes 48:58 intestinal γδ intraepithelial lymphocytes 18:58 intestinal γδ intraepithelial lymphocytes 18:58 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 5 83 theme core-1 912:917 arg1 structures 919:928 core-1 structures 912:928 core-1 structures 912:928 TCRδ(-/-) colons contained a lower proportion of core-2 structures and an increased proportion of core-1 structures whereas TCRδ(-/-) small intestines had a decreased percentage of core-3 structures. 25187161 2 84 theme γδ 479:480 arg1 TCRδ 488:491 TCRδ 488:491 TCRδ(-/-) 488:496 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 2 84 theme γδ 479:480 arg1 IELs 482:485 γδ IELs 479:485 γδ IELs (TCRδ(-/-)) 479:497 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 2 85 theme mice 466:469 arg1 profile 455:461 the intestinal O-glycosylation profile 424:461 the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 424:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 0 86 theme intraepithelial 32:46 arg1 lymphocytes 48:58 intestinal γδ intraepithelial lymphocytes 18:58 intestinal γδ intraepithelial lymphocytes 18:58 Mice deficient in intestinal γδ intraepithelial lymphocytes display an altered intestinal O-glycan profile compared with wild-type littermates. 25187161 1 87 theme mucosal 260:266 arg1 homeostasis 268:278 mucosal homeostasis 260:278 mucosal homeostasis 260:278 Intestinal γδ T-cell receptor-bearing intraepithelial lymphocytes (γδ IELs) play a multifaceted role in maintaining mucosal homeostasis. 25187161 2 88 theme profile 455:461 arg1 comparison 410:419 an in-depth mass spectrometric comparison 379:419 an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 379:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 7 89 dep were 1138:1141 arg1 whereas 1309:1315 whereas 1309:1315 whereas 1309:1315 There were significant differences in the sialylated species between the TCRδ(-/-) and WT mice with the sialylated Tn antigen found exclusively in the TCRδ(-/-)small intestine, whereas the sulfation pattern remained mostly unchanged. 25187161 3 90 theme TCRδ 667:670 arg1 intestine 644:652 small intestine 638:652 small intestine 638:652 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 3 90 theme TCRδ 667:670 arg1 colon 658:662 colon 658:662 colon 658:662 A total of 69 nonsulfated and 59 sulfated compositional types of O-glycans were identified in the small intestine and colon of TCRδ(-/-) and WT mice. 25187161 4 91 theme WT 798:799 arg1 littermates 801:811 WT littermates 798:811 WT littermates 798:811 Our results demonstrated structural differences in intestinal glycosylation in TCRδ(-/-) mice compared with WT littermates. 25187161 2 92 dep wild-type 512:520 arg1 WT 523:524 WT 523:524 WT 523:524 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 25187161 6 93 theme small 1057:1061 arg1 intestine 1063:1071 small intestine 1057:1071 small intestine 1057:1071 The glycan antennae in TCRδ(-/-) colon and small intestine showed altered structural diversity compared with WT mice. 25187161 2 94 theme spectrometric 396:408 arg1 comparison 410:419 an in-depth mass spectrometric comparison 379:419 an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates 379:537 In order to investigate the relationship between O-glycosylation and inflammation, we carried out an in-depth mass spectrometric comparison of the intestinal O-glycosylation profile of mice lacking γδ IELs (TCRδ(-/-)) and of their wild-type (WT) littermates. 26100261 2 0 theme reported 512:519 arg1 structures 529:538 three previously reported core M1 structures 495:538 three previously reported core M1 structures 495:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 2 1 theme M1 444:445 arg1 glycans 457:463 three more complex core M1 O-mannose glycans 420:463 three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures 420:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 2 theme prepared 272:279 arg1 intermediate 300:311 a chemically prepared disaccharyl serine intermediate 259:311 a chemically prepared disaccharyl serine intermediate 259:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 3 theme disaccharyl 281:291 arg1 intermediate 300:311 a chemically prepared disaccharyl serine intermediate 259:311 a chemically prepared disaccharyl serine intermediate 259:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 2 4 dep flexibility 323:333 arg1 The 314:316 The 314:316 The 314:316 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 5 gly glycosylation 242:254 arg1 intermediate 300:311 a chemically prepared disaccharyl serine intermediate 259:311 a chemically prepared disaccharyl serine intermediate 259:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 6 theme diversity-oriented 74:91 arg1 synthesis 108:116 The diversity-oriented chemoenzymatic synthesis 70:116 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans 70:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 7 theme chemoenzymatic 93:106 arg1 synthesis 108:116 The diversity-oriented chemoenzymatic synthesis 70:116 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans 70:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 2 8 theme first 473:477 arg1 time 479:482 the first time 469:482 the first time along with three previously reported core M1 structures 469:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 0 9 theme Chemoenzymatic 0:13 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans 0:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 1 10 theme serine 293:298 arg1 intermediate 300:311 a chemically prepared disaccharyl serine intermediate 259:311 a chemically prepared disaccharyl serine intermediate 259:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 11 theme three-step 193:202 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 0 12 theme α-dystroglycan 28:41 arg1 glycans 61:67 α-dystroglycan core M1 O-mannose glycans 28:67 α-dystroglycan core M1 O-mannose glycans 28:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 1 13 theme intermediate 300:311 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 14 theme α-dystroglycan 121:134 arg1 glycans 161:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 15 dep α-dystroglycan 121:134 arg1 α-DG 137:140 α-DG 137:140 α-DG 137:140 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 16 theme sequential 204:213 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 2 17 theme glycans 457:463 arg1 synthesis 407:415 the synthesis 403:415 the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures 403:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 2 18 theme high 318:321 arg1 flexibility 323:333 flexibility 323:333 flexibility 323:333 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 19 theme one-pot 215:221 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 0 20 theme M1 48:49 arg1 glycans 61:67 α-dystroglycan core M1 O-mannose glycans 28:67 α-dystroglycan core M1 O-mannose glycans 28:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 2 21 theme core 439:442 arg1 glycans 457:463 three more complex core M1 O-mannose glycans 420:463 three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures 420:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 22 theme multienzyme 223:233 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 0 23 theme core 43:46 arg1 glycans 61:67 α-dystroglycan core M1 O-mannose glycans 28:67 α-dystroglycan core M1 O-mannose glycans 28:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 2 24 theme complex 431:437 arg1 glycans 457:463 three more complex core M1 O-mannose glycans 420:463 three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures 420:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 0 25 theme glycans 61:67 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans 0:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 2 26 theme strategy 373:380 arg1 flexibility 323:333 flexibility 323:333 flexibility 323:333 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 2 26 theme strategy 373:380 arg1 efficiency 339:348 efficiency 339:348 efficiency 339:348 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 0 27 theme O-mannose 51:59 arg1 glycans 61:67 α-dystroglycan core M1 O-mannose glycans 28:67 α-dystroglycan core M1 O-mannose glycans 28:67 Chemoenzymatic synthesis of α-dystroglycan core M1 O-mannose glycans. 26100261 2 28 theme chemoenzymatic 358:371 arg1 strategy 373:380 this chemoenzymatic strategy 353:380 this chemoenzymatic strategy 353:380 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 29 theme core 143:146 arg1 glycans 161:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 30 theme OPME 236:239 arg1 glycosylation 242:254 a three-step sequential one-pot multienzyme (OPME) glycosylation 191:254 a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate 191:311 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 1 31 theme M1 148:149 arg1 glycans 161:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 2 32 theme M1 526:527 arg1 structures 529:538 three previously reported core M1 structures 495:538 three previously reported core M1 structures 495:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 2 33 theme O-mannose 447:455 arg1 glycans 457:463 three more complex core M1 O-mannose glycans 420:463 three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures 420:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 34 theme O-mannose 151:159 arg1 glycans 161:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 α-dystroglycan (α-DG) core M1 O-mannose glycans 121:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 26100261 2 35 theme core 521:524 arg1 structures 529:538 three previously reported core M1 structures 495:538 three previously reported core M1 structures 495:538 The high flexibility and efficiency of this chemoenzymatic strategy was demonstrated for the synthesis of three more complex core M1 O-mannose glycans for the first time along with three previously reported core M1 structures. 26100261 1 36 theme glycans 161:167 arg1 synthesis 108:116 The diversity-oriented chemoenzymatic synthesis 70:116 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans 70:167 The diversity-oriented chemoenzymatic synthesis of α-dystroglycan (α-DG) core M1 O-mannose glycans has been achieved via a three-step sequential one-pot multienzyme (OPME) glycosylation of a chemically prepared disaccharyl serine intermediate. 25652217 4 0 theme side 705:708 arg1 chain 710:714 the Trp168 side chain 694:714 the Trp168 side chain 694:714 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 7 1 theme TG 1183:1184 arg1 activity 1186:1193 the efficient TG activity 1169:1193 the efficient TG activity 1169:1193 These structural features of CrChiA appear to be responsible for the efficient TG activity. 25652217 10 2 theme V 1825:1825 arg1 chitinases 1827:1836 plant class V chitinases 1813:1836 plant class V chitinases 1813:1836 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 4 3 dep site 662:665 arg1 +3 679:680 +3 679:680 +3 679:680 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 3 dep site 662:665 arg1 +2 672:673 +2 672:673 +2 672:673 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 3 dep site 662:665 arg1 +1 668:669 +1 668:669 +1 668:669 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 9 4 from those 1526:1530 arg1 different 1511:1519 different 1511:1519 different 1511:1519 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 4 5 theme CrChiA 686:691 arg1 site 662:665 the acceptor binding site 641:665 the acceptor binding site (+1, +2 and +3) of CrChiA 641:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 5 theme CrChiA 686:691 arg1 CrChiA 686:691 CrChiA 686:691 CrChiA 686:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 6 6 theme middle 934:939 arg1 Asp117 946:951 Asp117 946:951 Asp117 946:951 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 6 theme middle 934:939 arg1 Asp 941:943 the middle Asp 930:943 the middle Asp (Asp117) 930:952 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 10 7 theme plant 1813:1817 arg1 chitinases 1827:1836 plant class V chitinases 1813:1836 plant class V chitinases 1813:1836 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 1 8 theme hydrolase 171:179 arg1 family 181:186 glycoside hydrolase family 18 161:189 glycoside hydrolase family 18 161:189 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 8 theme hydrolase 171:179 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 0 9 theme unique 107:112 arg1 features 140:147 unique structural and functional features 107:147 unique structural and functional features 107:147 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 2 10 theme Arabidopsis 468:478 arg1 thaliana 480:487 Arabidopsis thaliana 468:487 Arabidopsis thaliana (AtChiC) 468:496 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 2 10 theme Arabidopsis 468:478 arg1 AtChiC 490:495 AtChiC 490:495 AtChiC 490:495 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 1 11 dep Cycas 217:221 arg1 CrChiA 233:238 CrChiA 233:238 CrChiA 233:238 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 11 dep Cycas 217:221 arg1 revoluta 223:230 revoluta 223:230 the cycad Cycas revoluta (CrChiA) 207:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 7 12 theme CrChiA 1133:1138 arg1 features 1121:1128 These structural features 1104:1128 These structural features of CrChiA 1104:1138 These structural features of CrChiA appear to be responsible for the efficient TG activity. 25652217 3 13 contain had 571:573 arg1 chitinases 560:569 All three chitinases 550:569 All three chitinases 550:569 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 3 13 contain had 571:573 arg2 fold 599:602 a similar (α/β)8 barrel fold 575:602 a similar (α/β)8 barrel fold with an (α + β) insertion domain 575:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 6 14 theme catalytic 985:993 arg1 Glu119 1000:1005 the catalytic acid Glu119 981:1005 the catalytic acid Glu119 in CrChiA 981:1015 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 9 15 theme TG 1657:1658 arg1 activity 1660:1667 significant TG activity 1645:1667 significant TG activity 1645:1667 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 1 16 from Cycas 217:221 arg1 chitinase 252:260 a plant chitinase 244:260 a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity 244:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 16 from Cycas 217:221 arg1 family 181:186 glycoside hydrolase family 18 161:189 glycoside hydrolase family 18 161:189 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 16 from Cycas 217:221 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 9 17 theme solvation 1442:1450 arg1 changes 1479:1485 solvation and conformational entropy changes 1442:1485 solvation and conformational entropy changes in CrChiA 1442:1495 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 2 18 from tabacum 447:453 arg1 chitinases 421:430 class V chitinases 413:430 class V chitinases 413:430 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 2 19 theme Nicotiana 437:445 arg1 NtChiV 456:461 NtChiV 456:461 NtChiV 456:461 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 2 19 theme Nicotiana 437:445 arg1 tabacum 447:453 Nicotiana tabacum 437:453 Nicotiana tabacum (NtChiV) 437:462 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 8 20 theme allosamidin 1226:1236 arg1 binding 1201:1207 binding 1201:1207 binding of the inhibitor allosamidin 1201:1236 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 9 21 from marcescens 1604:1613 arg1 A 1588:1588 chitinase A 1578:1588 chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity 1578:1667 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 8 22 theme kcal 1418:1421 arg1 -1 1427:1428 -1 1427:1428 -1 1427:1428 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 8 22 theme kcal 1418:1421 arg1 mol 1423:1425 -9.8 kcal mol 1413:1425 -9.8 kcal mol(-1) 1413:1429 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 0 23 theme class 61:65 arg1 chitinases 69:78 plant class V chitinases 55:78 plant class V chitinases 55:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 8 24 dep other 1384:1388 arg1 i.e. 1391:1394 i.e. 1391:1394 i.e. 1391:1394 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 0 25 theme chitinases 69:78 arg1 properties 41:50 inhibitor binding properties 23:50 inhibitor binding properties of plant class V chitinases 23:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 0 25 theme chitinases 69:78 arg1 structures 8:17 Crystal structures 0:17 Crystal structures 0:17 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 1 26 theme efficient 296:304 arg1 activity 330:337 efficient transglycosylation (TG) activity 296:337 efficient transglycosylation (TG) activity 296:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 27 theme class 152:156 arg1 chitinase 252:260 a plant chitinase 244:260 a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity 244:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 27 theme class 152:156 arg1 family 181:186 glycoside hydrolase family 18 161:189 glycoside hydrolase family 18 161:189 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 27 theme class 152:156 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 8 28 theme isothermal 1258:1267 arg1 calorimetry 1279:1289 isothermal titration calorimetry 1258:1289 isothermal titration calorimetry 1258:1289 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 3 29 dep domain 630:635 arg1 β 617:617 β 617:617 β 617:617 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 3 29 dep domain 630:635 arg1 α 613:613 α 613:613 α 613:613 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 4 30 with face-to-face 735:746 arg1 sugar 760:764 the +3 sugar 753:764 the +3 sugar 753:764 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 9 31 theme conformational 1456:1469 arg1 entropy 1471:1477 conformational entropy 1456:1477 conformational entropy 1456:1477 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 0 32 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures 0:17 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 2 33 theme CrChiA 375:380 arg1 structure 362:370 the crystal structure 350:370 the crystal structure of CrChiA 350:380 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 1 34 theme plant 246:250 arg1 chitinase 252:260 a plant chitinase 244:260 a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity 244:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 34 theme plant 246:250 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 2 35 theme class 413:417 arg1 chitinases 421:430 class V chitinases 413:430 class V chitinases 413:430 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 8 36 theme free 1315:1318 arg1 energy 1320:1325 binding free energy 1307:1325 binding free energy of the three chitinases 1307:1349 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 0 37 theme binding 33:39 arg1 properties 41:50 inhibitor binding properties 23:50 inhibitor binding properties of plant class V chitinases 23:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 9 38 from changes 1479:1485 arg1 CrChiA 1490:1495 CrChiA 1490:1495 CrChiA 1490:1495 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 5 39 theme NtChiV 835:840 arg1 regions 824:830 the identical regions 810:830 the identical regions of NtChiV and AtChiC 810:851 However, this interaction was not found in the identical regions of NtChiV and AtChiC. 25652217 3 40 theme α/β 586:588 arg1 fold 599:602 a similar (α/β)8 barrel fold 575:602 a similar (α/β)8 barrel fold with an (α + β) insertion domain 575:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 4 41 theme binding 654:660 arg1 site 662:665 the acceptor binding site 641:665 the acceptor binding site (+1, +2 and +3) of CrChiA 641:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 41 theme binding 654:660 arg1 CrChiA 686:691 CrChiA 686:691 CrChiA 686:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 1 42 contain possess 288:294 arg2 activity 330:337 efficient transglycosylation (TG) activity 296:337 efficient transglycosylation (TG) activity 296:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 42 contain possess 288:294 arg1 chitinase 252:260 a plant chitinase 244:260 a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity 244:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 42 contain possess 288:294 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 7 43 theme efficient 1173:1181 arg1 activity 1186:1193 the efficient TG activity 1169:1193 the efficient TG activity 1169:1193 These structural features of CrChiA appear to be responsible for the efficient TG activity. 25652217 3 44 theme similar 577:583 arg1 fold 599:602 a similar (α/β)8 barrel fold 575:602 a similar (α/β)8 barrel fold with an (α + β) insertion domain 575:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 1 45 theme cycad 211:215 arg1 Cycas 217:221 the cycad Cycas revoluta (CrChiA) 207:239 the cycad Cycas revoluta (CrChiA) 207:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 9 46 theme chitinase 1578:1586 arg1 A 1588:1588 chitinase A 1578:1588 chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity 1578:1667 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 2 47 from thaliana 480:487 arg1 chitinases 421:430 class V chitinases 413:430 class V chitinases 413:430 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 5 48 located found 801:805 arg1 regions 824:830 the identical regions 810:830 the identical regions of NtChiV and AtChiC 810:851 However, this interaction was not found in the identical regions of NtChiV and AtChiC. 25652217 5 48 located found 801:805 arg2 interaction 781:791 this interaction 776:791 this interaction 776:791 However, this interaction was not found in the identical regions of NtChiV and AtChiC. 25652217 2 49 theme TG 537:538 arg1 reaction 540:547 the TG reaction 533:547 the TG reaction 533:547 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 10 50 theme bacterial 1789:1797 arg1 chitinases 1799:1808 bacterial chitinases 1789:1808 bacterial chitinases 1789:1808 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 6 51 theme corresponding 1030:1042 arg1 Asp 1044:1046 the corresponding Asp 1026:1046 the corresponding Asp in NtChiV and AtChiC 1026:1067 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 3 52 with fold 599:602 arg1 domain 630:635 an (α + β) insertion domain 609:635 an (α + β) insertion domain 609:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 4 53 theme Trp168 698:703 arg1 chain 710:714 the Trp168 side chain 694:714 the Trp168 side chain 694:714 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 1 54 theme V 158:158 arg1 chitinase 252:260 a plant chitinase 244:260 a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity 244:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 54 theme V 158:158 arg1 family 181:186 glycoside hydrolase family 18 161:189 glycoside hydrolase family 18 161:189 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 54 theme V 158:158 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 6 55 theme Asp 941:943 arg1 group 921:925 the carboxyl group 908:925 the carboxyl group of the middle Asp (Asp117) 908:952 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 55 theme Asp 941:943 arg1 Asp117 946:951 Asp117 946:951 Asp117 946:951 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 55 theme Asp 941:943 arg1 Asp 941:943 the middle Asp 930:943 the middle Asp (Asp117) 930:952 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 10 56 theme class 1819:1823 arg1 chitinases 1827:1836 plant class V chitinases 1813:1836 plant class V chitinases 1813:1836 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 1 57 theme glycoside 161:169 arg1 family 181:186 glycoside hydrolase family 18 161:189 glycoside hydrolase family 18 161:189 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 57 theme glycoside 161:169 arg1 chitinase 192:200 A class V (glycoside hydrolase family 18) chitinase 150:200 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) 150:239 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 3 58 theme insertion 620:628 arg1 domain 630:635 an (α + β) insertion domain 609:635 an (α + β) insertion domain 609:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 6 59 theme first 1093:1097 arg1 Asp 1099:1101 the first Asp 1089:1101 the first Asp 1089:1101 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 4 60 located found 720:724 arg1 site 662:665 the acceptor binding site 641:665 the acceptor binding site (+1, +2 and +3) of CrChiA 641:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 60 located found 720:724 arg2 chain 710:714 the Trp168 side chain 694:714 the Trp168 side chain 694:714 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 60 located found 720:724 arg1 CrChiA 686:691 CrChiA 686:691 CrChiA 686:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 0 61 theme functional 129:138 arg1 features 140:147 unique structural and functional features 107:147 unique structural and functional features 107:147 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 6 62 theme acid 995:998 arg1 Glu119 1000:1005 the catalytic acid Glu119 981:1005 the catalytic acid Glu119 in CrChiA 981:1015 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 9 63 theme significant 1645:1655 arg1 activity 1660:1667 significant TG activity 1645:1667 significant TG activity 1645:1667 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 0 64 theme structural 114:123 arg1 features 140:147 unique structural and functional features 107:147 unique structural and functional features 107:147 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 8 65 theme inhibitor 1216:1224 arg1 allosamidin 1226:1236 the inhibitor allosamidin 1212:1236 the inhibitor allosamidin 1212:1236 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 5 66 theme AtChiC 846:851 arg1 regions 824:830 the identical regions 810:830 the identical regions of NtChiV and AtChiC 810:851 However, this interaction was not found in the identical regions of NtChiV and AtChiC. 25652217 7 67 theme structural 1110:1119 arg1 features 1121:1128 These structural features 1104:1128 These structural features of CrChiA 1104:1138 These structural features of CrChiA appear to be responsible for the efficient TG activity. 25652217 6 68 from Glu119 1000:1005 arg1 CrChiA 1010:1015 CrChiA 1010:1015 CrChiA 1010:1015 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 10 69 theme molecular 1709:1717 arg1 mechanism 1719:1727 the molecular mechanism 1705:1727 the molecular mechanism underlying the TG reaction 1705:1754 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 0 70 theme V 67:67 arg1 chitinases 69:78 plant class V chitinases 55:78 plant class V chitinases 55:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 9 71 theme entropy 1471:1477 arg1 changes 1479:1485 solvation and conformational entropy changes 1442:1485 solvation and conformational entropy changes in CrChiA 1442:1495 However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. 25652217 2 72 theme V 419:419 arg1 chitinases 421:430 class V chitinases 413:430 class V chitinases 413:430 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 8 73 theme titration 1269:1277 arg1 calorimetry 1279:1289 isothermal titration calorimetry 1258:1289 isothermal titration calorimetry 1258:1289 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 10 74 theme TG 1744:1745 arg1 reaction 1747:1754 the TG reaction 1740:1754 the TG reaction 1740:1754 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 0 75 theme cycad 85:89 arg1 enzyme 91:96 the cycad enzyme 81:96 the cycad enzyme 81:96 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 1 76 theme transglycosylation 306:323 arg1 activity 330:337 efficient transglycosylation (TG) activity 296:337 efficient transglycosylation (TG) activity 296:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 1 77 theme TG 326:327 arg1 activity 330:337 efficient transglycosylation (TG) activity 296:337 efficient transglycosylation (TG) activity 296:337 A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. 25652217 6 78 theme DxDxE 861:865 arg1 essential 883:891 essential 883:891 essential 883:891 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 78 theme DxDxE 861:865 arg1 motif 867:871 the DxDxE motif 857:871 the DxDxE motif 857:871 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 8 79 theme binding 1307:1313 arg1 energy 1320:1325 binding free energy 1307:1325 binding free energy of the three chitinases 1307:1349 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 0 80 theme inhibitor 23:31 arg1 properties 41:50 inhibitor binding properties 23:50 inhibitor binding properties of plant class V chitinases 23:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 10 81 from chitinases 1799:1808 arg1 evolution 1774:1782 the molecular evolution 1760:1782 the molecular evolution from bacterial chitinases to plant class V chitinases 1760:1836 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 10 81 from chitinases 1799:1808 arg1 insight 1692:1698 insight 1692:1698 insight into the molecular mechanism underlying the TG reaction 1692:1754 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 8 82 from changes 1296:1302 arg1 energy 1320:1325 binding free energy 1307:1325 binding free energy of the three chitinases 1307:1349 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 5 83 theme identical 814:822 arg1 regions 824:830 the identical regions 810:830 the identical regions of NtChiV and AtChiC 810:851 However, this interaction was not found in the identical regions of NtChiV and AtChiC. 25652217 2 84 theme crystal 354:360 arg1 structure 362:370 the crystal structure 350:370 the crystal structure of CrChiA 350:380 We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. 25652217 0 85 theme plant 55:59 arg1 chitinases 69:78 plant class V chitinases 55:78 plant class V chitinases 55:78 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 6 86 theme carboxyl 912:919 arg1 group 921:925 the carboxyl group 908:925 the carboxyl group of the middle Asp (Asp117) 908:952 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 86 theme carboxyl 912:919 arg1 Asp117 946:951 Asp117 946:951 Asp117 946:951 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 86 theme carboxyl 912:919 arg1 Asp 941:943 the middle Asp 930:943 the middle Asp (Asp117) 930:952 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 8 87 theme chitinases 1340:1349 arg1 energy 1320:1325 binding free energy 1307:1325 binding free energy of the three chitinases 1307:1349 When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between -9.5 and -9.8 kcal mol(-1) . 25652217 10 88 theme molecular 1764:1772 arg1 evolution 1774:1782 the molecular evolution 1760:1782 the molecular evolution from bacterial chitinases to plant class V chitinases 1760:1836 These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases. 25652217 4 89 theme acceptor 645:652 arg1 site 662:665 the acceptor binding site 641:665 the acceptor binding site (+1, +2 and +3) of CrChiA 641:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 4 89 theme acceptor 645:652 arg1 CrChiA 686:691 CrChiA 686:691 CrChiA 686:691 In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face-to-face with the +3 sugar. 25652217 0 90 dep structures 8:17 arg1 exhibits 98:105 exhibits 98:105 exhibits unique structural and functional features 98:147 Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features. 25652217 3 91 theme barrel 592:597 arg1 fold 599:602 a similar (α/β)8 barrel fold 575:602 a similar (α/β)8 barrel fold with an (α + β) insertion domain 575:635 All three chitinases had a similar (α/β)8 barrel fold with an (α + β) insertion domain. 25652217 6 92 from Asp 1044:1046 arg1 AtChiC 1062:1067 AtChiC 1062:1067 AtChiC 1062:1067 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 25652217 6 92 from Asp 1044:1046 arg1 NtChiV 1051:1056 NtChiV 1051:1056 NtChiV 1051:1056 In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. 26527586 5 0 theme aromatic 772:779 arg1 likely 797:802 likely 797:802 likely 797:802 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 5 0 theme aromatic 772:779 arg1 surface 781:787 a hydrophobic aromatic surface 758:787 a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors 758:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 3 1 dep 97  563:565 arg1 to 560:561 to 560:561 to 560:561 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 0 2 theme Clash-Induced 63:75 arg1 Cascade 77:83 a Clash-Induced Cascade 61:83 a Clash-Induced Cascade of Loop Shifts 61:98 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 1 3 theme enzymatic 149:157 arg1 production 159:168 the enzymatic production 145:168 the enzymatic production of glycosylated compounds for decades 145:206 Sucrose phosphorylases have been applied in the enzymatic production of glycosylated compounds for decades. 26527586 3 4 from yields 547:552 arg1 -epicatechin 531:542 -epicatechin 531:542 -epicatechin 531:542 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 0 5 theme Loop 88:91 arg1 Shifts 93:98 Loop Shifts 88:98 Loop Shifts 88:98 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 5 6 theme access 738:743 arg1 channel 745:751 a widened access channel 728:751 a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors 728:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 1 7 gly glycosylated 173:184 arg1 compounds 186:194 glycosylated compounds 173:194 glycosylated compounds for decades 173:206 Sucrose phosphorylases have been applied in the enzymatic production of glycosylated compounds for decades. 26527586 6 8 theme Q345F 980:984 arg1 exchange 986:993 the Q345F exchange 976:993 the Q345F exchange 976:993 The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange. 26527586 0 9 theme Shifts 93:98 arg1 Cascade 77:83 a Clash-Induced Cascade 61:83 a Clash-Induced Cascade of Loop Shifts 61:98 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 6 10 theme channel 894:900 arg1 generation 875:884 The generation 871:884 The generation of this channel 871:900 The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange. 26527586 7 11 theme other 1064:1068 arg1 enzymes 1082:1088 other tailor-made enzymes 1064:1088 other tailor-made enzymes 1064:1088 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 3 12 theme efficient 471:479 arg1 glucosylation 481:493 efficient glucosylation 471:493 efficient glucosylation 471:493 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 3 13 theme Q345F 390:394 arg1 exchange 396:403 The Q345F exchange 386:403 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis 386:462 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 3 14 from exchange 396:403 arg1 phosphorylase 416:428 sucrose phosphorylase 408:428 sucrose phosphorylase 408:428 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 7 15 theme tailor-made 1070:1080 arg1 enzymes 1082:1088 other tailor-made enzymes 1064:1088 other tailor-made enzymes 1064:1088 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 3 16 theme sucrose 408:414 arg1 phosphorylase 416:428 sucrose phosphorylase 408:428 sucrose phosphorylase 408:428 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 1 17 theme glycosylated 173:184 arg1 compounds 186:194 glycosylated compounds 173:194 glycosylated compounds for decades 173:206 Sucrose phosphorylases have been applied in the enzymatic production of glycosylated compounds for decades. 26527586 7 18 theme enzymes 1082:1088 arg1 design 1054:1059 the design 1050:1059 the design of other tailor-made enzymes 1050:1088 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 2 19 theme antimicrobial 303:315 arg1 properties 350:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 1 20 theme compounds 186:194 arg1 production 159:168 the enzymatic production 145:168 the enzymatic production of glycosylated compounds for decades 145:206 Sucrose phosphorylases have been applied in the enzymatic production of glycosylated compounds for decades. 26527586 0 21 theme Active 16:21 arg1 Site 23:26 the Active Site 12:26 the Active Site of Sucrose Phosphorylase 12:51 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 0 21 theme Active 16:21 arg1 Phosphorylase 39:51 Sucrose Phosphorylase 31:51 Sucrose Phosphorylase 31:51 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 2 22 theme diverse 295:301 arg1 properties 350:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 5 23 with channel 745:751 arg1 likely 797:802 likely 797:802 likely 797:802 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 5 23 with channel 745:751 arg1 surface 781:787 a hydrophobic aromatic surface 758:787 a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors 758:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 3 24 theme resveratrol 498:508 arg1 glucosylation 481:493 efficient glucosylation 471:493 efficient glucosylation 471:493 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 3 25 theme % 566:566 arg1 yields 547:552 yields 547:552 yields of up to 97 % 547:566 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 5 26 theme variant 712:718 arg1 structure 695:703 The crystal structure 683:703 The crystal structure of the variant 683:718 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 6 27 theme changes 955:961 arg1 cascade 933:939 a cascade 931:939 a cascade of structural changes arising from the Q345F exchange 931:993 The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange. 26527586 3 28 theme wild-type 580:588 arg1 enzyme 590:595 the wild-type enzyme 576:595 the wild-type enzyme 576:595 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 5 29 theme aromatic 851:858 arg1 acceptors 860:868 aromatic acceptors 851:868 aromatic acceptors 851:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 0 30 theme Site 23:26 arg1 Redesign 0:7 Redesign 0:7 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 0:99 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 6 31 theme structural 944:953 arg1 changes 955:961 structural changes 944:961 structural changes arising from the Q345F exchange 944:993 The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange. 26527586 2 32 theme desirable 226:234 arg1 acceptors 236:244 several desirable acceptors 218:244 several desirable acceptors 218:244 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 2 32 theme desirable 226:234 arg1 stilbenoids 269:279 stilbenoids 269:279 stilbenoids 269:279 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 2 32 theme desirable 226:234 arg1 flavonoids 255:264 flavonoids 255:264 flavonoids 255:264 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 4 33 theme undescribed 642:652 arg1 products 654:661 Three previously undescribed products 625:661 Three previously undescribed products 625:661 Three previously undescribed products are made available. 26527586 0 34 theme Phosphorylase 39:51 arg1 Site 23:26 the Active Site 12:26 the Active Site of Sucrose Phosphorylase 12:51 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 0 34 theme Phosphorylase 39:51 arg1 Phosphorylase 39:51 Sucrose Phosphorylase 31:51 Sucrose Phosphorylase 31:51 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 5 35 theme widened 730:736 arg1 channel 745:751 a widened access channel 728:751 a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors 728:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 2 36 theme several 218:224 arg1 acceptors 236:244 several desirable acceptors 218:244 several desirable acceptors 218:244 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 2 36 theme several 218:224 arg1 stilbenoids 269:279 stilbenoids 269:279 stilbenoids 269:279 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 2 36 theme several 218:224 arg1 flavonoids 255:264 flavonoids 255:264 flavonoids 255:264 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 1 37 theme Sucrose 101:107 arg1 phosphorylases 109:122 Sucrose phosphorylases 101:122 Sucrose phosphorylases 101:122 Sucrose phosphorylases have been applied in the enzymatic production of glycosylated compounds for decades. 26527586 0 38 theme Sucrose 31:37 arg1 Phosphorylase 39:51 Sucrose Phosphorylase 31:51 Sucrose Phosphorylase 31:51 Redesign of the Active Site of Sucrose Phosphorylase through a Clash-Induced Cascade of Loop Shifts. 26527586 6 39 theme cascade 933:939 arg1 terms 922:926 terms 922:926 terms of a cascade of structural changes arising from the Q345F exchange 922:993 The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange. 26527586 5 40 theme crystal 687:693 arg1 structure 695:703 The crystal structure 683:703 The crystal structure of the variant 683:718 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 7 41 theme observed 1000:1007 arg1 relevant 1037:1044 relevant 1037:1044 relevant 1037:1044 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 7 41 theme observed 1000:1007 arg1 mechanisms 1009:1018 The observed mechanisms 996:1018 The observed mechanisms 996:1018 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 7 41 theme observed 1000:1007 arg1 likely 1024:1029 likely 1024:1029 likely 1024:1029 The observed mechanisms are likely to be relevant for the design of other tailor-made enzymes. 26527586 2 42 theme antioxidant 338:348 arg1 properties 350:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 5 43 theme improved 825:832 arg1 activity 834:841 the improved activity 821:841 the improved activity towards aromatic acceptors 821:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 2 44 theme anticarcinogenic 318:333 arg1 properties 350:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 diverse antimicrobial, anticarcinogenic or antioxidant properties 295:359 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 3 45 from adolescentis 451:462 arg1 exchange 396:403 The Q345F exchange 386:403 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis 386:462 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 5 46 theme hydrophobic 760:770 arg1 likely 797:802 likely 797:802 likely 797:802 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 5 46 theme hydrophobic 760:770 arg1 surface 781:787 a hydrophobic aromatic surface 758:787 a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors 758:868 The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. 26527586 3 47 theme sucrose 605:611 arg1 hydrolysis 613:622 sucrose hydrolysis 605:622 sucrose hydrolysis 605:622 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 26527586 2 48 theme poor 369:372 arg1 substrates 374:383 poor substrates 369:383 poor substrates 369:383 However, several desirable acceptors, such as flavonoids or stilbenoids, that exhibit diverse antimicrobial, anticarcinogenic or antioxidant properties, remain poor substrates. 26527586 3 49 from -epicatechin 531:542 arg1 yields 547:552 yields 547:552 yields of up to 97 % 547:566 The Q345F exchange in sucrose phosphorylase from Bifidobacterium adolescentis allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97 % whereas the wild-type enzyme favours sucrose hydrolysis. 29197583 4 0 theme glycoconjugates 798:812 arg1 separation 775:784 the separation 771:784 the separation of isomeric glycoconjugates 771:812 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 8 1 theme acid 1549:1552 arg1 linkage 1554:1560 sialic acid linkage 1542:1560 sialic acid linkage 1542:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 2 2 theme multiple 560:567 arg1 isomers 569:575 multiple isomers 560:575 multiple isomers 560:575 Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. 29197583 6 3 theme Straightforward 1092:1106 arg1 separation 1108:1117 Straightforward separation 1092:1117 Straightforward separation of isomers 1092:1128 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 0 4 theme glycan 164:169 arg1 level 171:175 glycan level 164:175 glycan level 164:175 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 8 5 dep SIGNIFICANCE 1402:1413 arg1 spectrometry 1433:1444 Ion mobility mass spectrometry 1415:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 7 6 theme real 1369:1372 arg1 samples 1382:1388 real complex samples 1369:1388 real complex samples 1369:1388 Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. 29197583 6 7 theme glycans 1173:1179 arg1 analysis 1152:1159 the analysis 1148:1159 the analysis of released glycans 1148:1179 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 9 8 theme modification 1590:1601 arg1 reason 1664:1669 reason 1664:1669 reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides 1664:1765 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 8 theme modification 1590:1601 arg1 evident 1635:1641 evident 1635:1641 evident 1635:1641 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 8 theme modification 1590:1601 arg1 impact 1567:1572 The impact 1563:1572 The impact of such a small modification in the glycan structure 1563:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 10 9 theme glycoconjugates 1870:1884 arg1 separation 1839:1848 the separation 1835:1848 the separation of highly decorated glycoconjugates 1835:1884 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 4 10 theme isomeric 789:796 arg1 glycoconjugates 798:812 isomeric glycoconjugates 789:812 isomeric glycoconjugates 789:812 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 7 11 theme serum 1301:1305 arg1 samples 1307:1313 serum samples 1301:1313 serum samples of mice 1301:1321 Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. 29197583 3 12 theme compounds 692:700 arg1 separation 669:678 the separation 665:678 the separation of isomeric compounds 665:700 In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. 29197583 0 13 theme acid 105:108 arg1 linkage 110:116 sialic acid linkage 98:116 sialic acid linkage 98:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 8 14 theme sialic 1542:1547 arg1 linkage 1554:1560 sialic acid linkage 1542:1560 sialic acid linkage 1542:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 1 15 theme type 357:360 arg1 task 264:267 an intricate but very important task 232:267 an intricate but very important task 232:267 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 15 theme type 357:360 arg1 glycomics 277:285 making glycomics one 270:289 making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science 270:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 16 dep challenging 303:313 arg1 under-researched 339:354 under-researched 339:354 under-researched 339:354 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 16 dep challenging 303:313 arg1 albeit 332:337 albeit 332:337 albeit 332:337 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 11 17 dep topic 1908:1912 arg1 nowadays 1914:1921 nowadays 1914:1921 nowadays 1914:1921 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 9 18 from impact 1567:1572 arg1 structure 1617:1625 the glycan structure 1606:1625 the glycan structure 1606:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 11 19 theme isomers 1971:1977 arg1 abundance 1946:1954 the abundance 1942:1954 the abundance of some glycan isomers 1942:1977 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 1 20 theme " 371:371 arg1 type 357:360 the most challenging and interesting, albeit under-researched, type 294:360 the most challenging and interesting, albeit under-researched, type of "omics" science 294:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 2 21 theme branched 468:475 arg1 structures 477:486 severely branched structures 459:486 severely branched structures 459:486 Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. 29197583 5 22 gly glycoprotein 1047:1058 arg1 glycoprotein 1047:1058 the intact glycoprotein 1036:1058 the intact glycoprotein 1036:1058 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 0 23 theme glycoconjugate 49:62 arg1 isomers 64:70 glycoconjugate isomers 49:70 glycoconjugate isomers 49:70 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 24 from evident 1635:1641 arg1 analytes 1654:1661 smaller analytes 1646:1661 smaller analytes 1646:1661 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 5 25 gly glycopeptide 1061:1072 arg2 glycopeptide 1061:1072 glycopeptide 1061:1072 glycopeptide 1061:1072 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 5 26 theme isomeric 968:975 arg1 glycoconjugates 977:991 isomeric glycoconjugates 968:991 isomeric glycoconjugates 968:991 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 27 gly glycopeptides 1753:1765 arg2 glycopeptides 1753:1765 the glycopeptides 1749:1765 the glycopeptides 1749:1765 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 2 28 theme different 498:506 arg1 constituents 508:519 many different constituents 493:519 many different constituents 493:519 Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. 29197583 8 29 theme mobility 1419:1426 arg1 spectrometry 1433:1444 Ion mobility mass spectrometry 1415:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 0 30 theme different 79:87 arg1 types 89:93 different types 79:93 different types of sialic acid linkage 79:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 6 31 theme isomers 1122:1128 arg1 separation 1108:1117 Straightforward separation 1092:1117 Straightforward separation of isomers 1092:1128 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 1 32 theme intricate 235:243 arg1 study 182:186 The study 178:186 The study of protein glycosylation 178:211 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 32 theme intricate 235:243 arg1 task 264:267 an intricate but very important task 232:267 an intricate but very important task 232:267 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 32 theme intricate 235:243 arg1 glycomics 277:285 making glycomics one 270:289 making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science 270:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 5 33 theme acid 1019:1022 arg1 linkage 1024:1030 sialic acid linkage 1012:1030 sialic acid linkage 1012:1030 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 34 theme glycans 1696:1702 arg1 analysis 1679:1686 the analysis 1675:1686 the analysis of free glycans 1675:1702 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 4 35 theme mouse 835:839 arg1 mTf 854:856 mTf 854:856 mTf 854:856 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 4 35 theme mouse 835:839 arg1 transferrin 841:851 mouse transferrin 835:851 mouse transferrin (mTf) 835:857 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 5 36 theme glycan 1078:1083 arg1 level 1085:1089 glycan level 1078:1089 glycan level 1078:1089 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 4 37 gly glycoprotein 868:879 arg1 glycoprotein 868:879 model glycoprotein 862:879 model glycoprotein 862:879 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 8 38 theme analytical 1461:1470 arg1 SIGNIFICANCE 1402:1413 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 8 38 theme analytical 1461:1470 arg1 technique 1472:1480 a promising analytical technique 1449:1480 a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage 1449:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 0 39 theme ion 14:16 arg1 mobility 18:25 ion mobility 14:25 ion mobility 14:25 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 1 40 theme important 254:262 arg1 study 182:186 The study 178:186 The study of protein glycosylation 178:211 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 40 theme important 254:262 arg1 task 264:267 an intricate but very important task 232:267 an intricate but very important task 232:267 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 40 theme important 254:262 arg1 glycomics 277:285 making glycomics one 270:289 making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science 270:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 0 41 from glycopeptide 147:158 arg1 types 89:93 different types 79:93 different types of sialic acid linkage 79:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 3 42 theme IM 608:609 arg1 spectrometry 612:623 ion mobility (IM) spectrometry 594:623 ion mobility (IM) spectrometry 594:623 In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. 29197583 5 43 theme platform 937:944 arg1 performance 917:927 the performance 913:927 the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage 913:1030 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 44 theme intact 1731:1736 arg1 protein 1738:1744 the intact protein 1727:1744 the intact protein 1727:1744 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 3 45 theme mobility 598:605 arg1 spectrometry 612:623 ion mobility (IM) spectrometry 594:623 ion mobility (IM) spectrometry 594:623 In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. 29197583 8 46 theme glycoconjugate 1504:1517 arg1 isomers 1519:1525 glycoconjugate isomers 1504:1525 glycoconjugate isomers 1504:1525 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 1 47 theme protein 191:197 arg1 glycosylation 199:211 protein glycosylation 191:211 protein glycosylation 191:211 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 9 48 from analytes 1654:1661 arg1 reason 1664:1669 reason 1664:1669 reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides 1664:1765 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 48 from analytes 1654:1661 arg1 evident 1635:1641 evident 1635:1641 evident 1635:1641 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 48 from analytes 1654:1661 arg1 impact 1567:1572 The impact 1563:1572 The impact of such a small modification in the glycan structure 1563:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 4 49 theme wave 751:754 arg1 TWIMS 760:764 traveling wave IM (TWIMS) 741:765 traveling wave IM (TWIMS) 741:765 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 7 50 theme complex 1374:1380 arg1 samples 1382:1388 real complex samples 1369:1388 real complex samples 1369:1388 Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. 29197583 2 51 theme isomers 569:575 arg1 formation 547:555 the formation 543:555 the formation of multiple isomers 543:575 Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. 29197583 0 52 from glycoprotein 133:144 arg1 types 89:93 different types 79:93 different types of sialic acid linkage 79:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 8 53 theme linkage 1554:1560 arg1 type 1534:1537 type 1534:1537 type of sialic acid linkage 1534:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 3 54 theme isomeric 683:690 arg1 compounds 692:700 isomeric compounds 683:700 isomeric compounds 683:700 In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. 29197583 11 55 theme important 1898:1906 arg1 This 1887:1890 This 1887:1890 This 1887:1890 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 11 55 theme important 1898:1906 arg1 topic 1908:1912 an important topic nowadays 1895:1921 an important topic nowadays 1895:1921 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 0 56 gly glycopeptide 147:158 arg2 glycopeptide 147:158 glycopeptide 147:158 glycopeptide 147:158 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 57 theme small 1584:1588 arg1 modification 1590:1601 such a small modification 1577:1601 such a small modification in the glycan structure 1577:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 6 58 theme released 1164:1171 arg1 glycans 1173:1179 released glycans 1164:1179 released glycans 1164:1179 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 6 59 gly glycopeptides 1200:1212 arg2 glycopeptides 1200:1212 the glycopeptides 1196:1212 the glycopeptides which showed a more complex pattern 1196:1248 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 9 60 from modification 1590:1601 arg1 structure 1617:1625 the glycan structure 1606:1625 the glycan structure 1606:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 7 61 theme mice 1318:1321 arg1 samples 1307:1313 serum samples 1301:1313 serum samples of mice 1301:1321 Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. 29197583 10 62 theme decorated 1860:1868 arg1 glycoconjugates 1870:1884 highly decorated glycoconjugates 1853:1884 highly decorated glycoconjugates 1853:1884 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 0 63 theme linkage 110:116 arg1 types 89:93 different types 79:93 different types of sialic acid linkage 79:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 11 64 theme early 2004:2008 arg1 diagnosis 2010:2018 the early diagnosis 2000:2018 the early diagnosis 2000:2018 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 9 65 theme glycan 1610:1615 arg1 structure 1617:1625 the glycan structure 1606:1625 the glycan structure 1606:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 6 66 theme complex 1234:1240 arg1 pattern 1242:1248 a more complex pattern 1227:1248 a more complex pattern 1227:1248 Straightforward separation of isomers was achieved with the analysis of released glycans, as opposed to the glycopeptides which showed a more complex pattern. 29197583 10 67 from point 1826:1830 arg1 separation 1839:1848 the separation 1835:1848 the separation of highly decorated glycoconjugates 1835:1884 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 0 68 theme intact 126:131 arg1 glycoprotein 133:144 the intact glycoprotein 122:144 the intact glycoprotein 122:144 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 5 69 theme intact 1040:1045 arg1 glycoprotein 1047:1058 the intact glycoprotein 1036:1058 the intact glycoprotein 1036:1058 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 1 70 theme omics 366:370 arg1 " 371:371 "omics" 365:371 "omics" science 365:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 70 theme omics 366:370 arg1 science 373:379 science 373:379 science 373:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 9 71 theme smaller 1646:1652 arg1 analytes 1654:1661 smaller analytes 1646:1661 smaller analytes 1646:1661 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 8 72 theme BIOLOGICAL 1391:1400 arg1 SIGNIFICANCE 1402:1413 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 8 72 theme BIOLOGICAL 1391:1400 arg1 technique 1472:1480 a promising analytical technique 1449:1480 a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage 1449:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 7 73 theme developed 1264:1272 arg1 methodology 1274:1284 the developed methodology 1260:1284 the developed methodology 1260:1284 Finally, the developed methodology was applied to serum samples of mice, to investigate its robustness when analyzing real complex samples. 29197583 0 74 theme isomers 64:70 arg1 separation 35:44 the separation 31:44 the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level 31:175 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 5 75 theme glycoconjugates 977:991 arg1 separation 954:963 the separation 950:963 the separation of isomeric glycoconjugates due to the type of sialic acid linkage 950:1030 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 8 76 theme Ion 1415:1417 arg1 spectrometry 1433:1444 Ion mobility mass spectrometry 1415:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 2 77 theme many 493:496 arg1 constituents 508:519 many different constituents 493:519 many different constituents 493:519 Complexity escalates remarkably when considering that carbohydrates can form severely branched structures with many different constituents, which often leads to the formation of multiple isomers. 29197583 0 78 gly glycoprotein 133:144 arg1 glycoprotein 133:144 the intact glycoprotein 122:144 the intact glycoprotein 122:144 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 9 79 from structure 1617:1625 arg1 reason 1664:1669 reason 1664:1669 reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides 1664:1765 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 79 from structure 1617:1625 arg1 evident 1635:1641 evident 1635:1641 evident 1635:1641 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 79 from structure 1617:1625 arg1 impact 1567:1572 The impact 1563:1572 The impact of such a small modification in the glycan structure 1563:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 11 80 theme glycan 1964:1969 arg1 isomers 1971:1977 some glycan isomers 1959:1977 some glycan isomers 1959:1977 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 8 81 theme mass 1428:1431 arg1 spectrometry 1433:1444 Ion mobility mass spectrometry 1415:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 1 82 theme challenging 303:313 arg1 type 357:360 the most challenging and interesting, albeit under-researched, type 294:360 the most challenging and interesting, albeit under-researched, type of "omics" science 294:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 4 83 theme model 862:866 arg1 glycoprotein 868:879 model glycoprotein 862:879 model glycoprotein 862:879 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 4 84 theme present 710:716 arg1 work 718:721 the present work 706:721 the present work 706:721 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 0 85 theme sialic 98:103 arg1 linkage 110:116 sialic acid linkage 98:116 sialic acid linkage 98:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 1 86 theme interesting 319:329 arg1 type 357:360 the most challenging and interesting, albeit under-researched, type 294:360 the most challenging and interesting, albeit under-researched, type of "omics" science 294:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 9 87 theme free 1691:1694 arg1 glycans 1696:1702 free glycans 1691:1702 free glycans 1691:1702 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 8 88 theme promising 1451:1459 arg1 SIGNIFICANCE 1402:1413 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry 1391:1444 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 8 88 theme promising 1451:1459 arg1 technique 1472:1480 a promising analytical technique 1449:1480 a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage 1449:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 5 89 theme linkage 1024:1030 arg1 type 1004:1007 the type 1000:1007 the type of sialic acid linkage 1000:1030 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 5 90 theme sialic 1012:1017 arg1 linkage 1024:1030 sialic acid linkage 1012:1030 sialic acid linkage 1012:1030 Particularly, we aim to assess the performance of this platform for the separation of isomeric glycoconjugates due to the type of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 11 91 theme diseases 2059:2066 arg1 differentiation 2032:2046 differentiation 2032:2046 differentiation 2032:2046 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 11 91 theme diseases 2059:2066 arg1 control 2021:2027 control 2021:2027 control 2021:2027 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 11 91 theme diseases 2059:2066 arg1 diagnosis 2010:2018 the early diagnosis 2000:2018 the early diagnosis 2000:2018 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 0 92 from level 171:175 arg1 types 89:93 different types 79:93 different types of sialic acid linkage 79:116 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 0 93 theme mobility 18:25 arg1 Evaluation 0:9 Evaluation 0:9 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 0:176 Evaluation of ion mobility for the separation of glycoconjugate isomers due to different types of sialic acid linkage, at the intact glycoprotein, glycopeptide and glycan level. 29197583 3 94 theme ion 594:596 arg1 spectrometry 612:623 ion mobility (IM) spectrometry 594:623 ion mobility (IM) spectrometry 594:623 In this regard, ion mobility (IM) spectrometry has recently demonstrated its power for the separation of isomeric compounds. 29197583 4 95 theme IM 756:757 arg1 TWIMS 760:764 traveling wave IM (TWIMS) 741:765 traveling wave IM (TWIMS) 741:765 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 9 96 dep evident 1635:1641 arg1 reason 1664:1669 reason 1664:1669 reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides 1664:1765 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 96 dep evident 1635:1641 arg1 evident 1635:1641 evident 1635:1641 evident 1635:1641 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 9 96 dep evident 1635:1641 arg1 impact 1567:1572 The impact 1563:1572 The impact of such a small modification in the glycan structure 1563:1625 The impact of such a small modification in the glycan structure is more evident in smaller analytes, reason why the analysis of free glycans was easier compared to the intact protein or the glycopeptides. 29197583 1 97 theme making 270:275 arg1 task 264:267 an intricate but very important task 232:267 an intricate but very important task 232:267 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 97 theme making 270:275 arg1 glycomics 277:285 making glycomics one 270:289 making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science 270:379 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 8 98 theme isomers 1519:1525 arg1 separation 1490:1499 the separation 1486:1499 the separation of glycoconjugate isomers due to type of sialic acid linkage 1486:1560 BIOLOGICAL SIGNIFICANCE Ion mobility mass spectrometry is a promising analytical technique for the separation of glycoconjugate isomers due to type of sialic acid linkage. 29197583 4 99 theme TWIMS 760:764 arg1 potential 728:736 the potential 724:736 the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates 724:812 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 11 100 from differences 1927:1937 arg1 abundance 1946:1954 the abundance 1942:1954 the abundance of some glycan isomers 1942:1977 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 10 101 theme established 1772:1782 arg1 methodology 1784:1794 The established methodology 1768:1794 The established methodology 1768:1794 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 10 101 theme established 1772:1782 arg1 point 1826:1830 starting point 1817:1830 starting point in the separation of highly decorated glycoconjugates 1817:1884 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 1 102 theme glycosylation 199:211 arg1 task 264:267 an intricate but very important task 232:267 an intricate but very important task 232:267 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 1 102 theme glycosylation 199:211 arg1 study 182:186 The study 178:186 The study of protein glycosylation 178:211 The study of protein glycosylation can be regarded as an intricate but very important task, making glycomics one of the most challenging and interesting, albeit under-researched, type of "omics" science. 29197583 10 103 theme starting 1817:1824 arg1 methodology 1784:1794 The established methodology 1768:1794 The established methodology 1768:1794 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 10 103 theme starting 1817:1824 arg1 point 1826:1830 starting point 1817:1830 starting point in the separation of highly decorated glycoconjugates 1817:1884 The established methodology could be regarded as starting point in the separation of highly decorated glycoconjugates. 29197583 4 104 theme traveling 741:749 arg1 TWIMS 760:764 traveling wave IM (TWIMS) 741:765 traveling wave IM (TWIMS) 741:765 In the present work, the potential of traveling wave IM (TWIMS) for the separation of isomeric glycoconjugates was evaluated, using mouse transferrin (mTf) as model glycoprotein. 29197583 11 105 theme certain 2051:2057 arg1 diseases 2059:2066 certain diseases 2051:2066 certain diseases 2051:2066 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 11 105 theme certain 2051:2057 arg1 cancer 2093:2098 cancer 2093:2098 cancer 2093:2098 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 29197583 11 105 theme certain 2051:2057 arg1 inflammation 2077:2088 inflammation 2077:2088 inflammation 2077:2088 This is an important topic nowadays, as differences in the abundance of some glycan isomers could be the key for the early diagnosis, control or differentiation of certain diseases, such as inflammation or cancer. 26663535 6 0 contain had 874:876 arg1 ions 841:844 the HexNAc-derived m/z 126 and 144 oxonium ions 798:844 ions 841:844 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 6 0 contain had 874:876 arg1 m/z 817:819 the HexNAc-derived m/z 126 and 144 oxonium ions 798:844 m/z 817:819 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 6 0 contain had 874:876 arg2 structures 899:908 completely different structures 878:908 completely different structures 878:908 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 4 1 used used 628:631 arg2 patterns 610:617 Such patterns 605:617 Such patterns 605:617 Such patterns could be used to distinguish between glycopeptide structural isomers. 26663535 7 2 theme ions 1084:1087 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 2 theme ions 1084:1087 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 2 theme ions 1084:1087 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 1 3 theme cellular 185:192 arg1 processes 194:202 cellular processes 185:202 cellular processes 185:202 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 5 4 theme deuterium-labelled 723:740 arg1 glycopeptides 742:754 deuterium-labelled glycopeptides 723:754 deuterium-labelled glycopeptides 723:754 To acquire a mechanistic insight, deuterium-labelled glycopeptides were prepared and analysed. 26663535 0 5 theme Structure 111:119 arg1 Evidence 88:95 Evidence 88:95 Evidence of the Glycan Structure 88:119 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 6 6 link HexNAc-derived 802:815 arg1 m/z 817:819 the HexNAc-derived m/z 126 and 144 oxonium ions 798:844 m/z 817:819 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 4 7 theme structural 669:678 arg1 isomers 680:686 glycopeptide structural isomers 656:686 glycopeptide structural isomers 656:686 Such patterns could be used to distinguish between glycopeptide structural isomers. 26663535 0 8 theme Glycan 104:109 arg1 Structure 111:119 the Glycan Structure 100:119 the Glycan Structure 100:119 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 7 9 theme future 1211:1216 arg1 studies 1233:1239 future glycoproteomic studies 1211:1239 future glycoproteomic studies 1211:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 6 10 theme HexNAc-derived 802:815 arg1 m/z 817:819 the HexNAc-derived m/z 126 and 144 oxonium ions 798:844 m/z 817:819 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 3 11 theme ion 545:547 arg1 patterns 563:570 characteristically different HexNAc oxonium ion fragmentation patterns 501:570 characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure 501:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 7 12 theme important 1185:1193 arg1 information 1195:1205 important information 1185:1205 important information for future glycoproteomic studies 1185:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 6 13 link Hex-derived 967:977 arg1 ions 987:990 abundant Hex-derived oxonium ions 958:990 abundant Hex-derived oxonium ions 958:990 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 3 14 theme recent 478:483 arg1 study 485:489 a recent study 476:489 a recent study 476:489 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 5 15 gly glycopeptides 742:754 arg2 glycopeptides 742:754 deuterium-labelled glycopeptides 723:754 deuterium-labelled glycopeptides 723:754 To acquire a mechanistic insight, deuterium-labelled glycopeptides were prepared and analysed. 26663535 4 16 theme glycopeptide 656:667 arg1 isomers 680:686 glycopeptide structural isomers 656:686 glycopeptide structural isomers 656:686 Such patterns could be used to distinguish between glycopeptide structural isomers. 26663535 6 17 theme oxonium 979:985 arg1 ions 987:990 abundant Hex-derived oxonium ions 958:990 abundant Hex-derived oxonium ions 958:990 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 5 18 theme mechanistic 702:712 arg1 insight 714:720 a mechanistic insight 700:720 a mechanistic insight 700:720 To acquire a mechanistic insight, deuterium-labelled glycopeptides were prepared and analysed. 26663535 6 19 theme H2 868:869 arg1 O 871:871 H2 O 868:871 H2 O 868:871 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 2 20 theme pathways 454:461 arg1 assumption 417:426 the assumption 413:426 the assumption of conserved biosynthetic pathways 413:461 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 0 21 theme MS/MS 12:16 arg1 Pathways 32:39 Distinctive MS/MS Fragmentation Pathways 0:39 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions 0:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 6 22 theme Hex-derived 967:977 arg1 ions 987:990 abundant Hex-derived oxonium ions 958:990 abundant Hex-derived oxonium ions 958:990 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 2 23 theme biosynthetic 441:452 arg1 pathways 454:461 conserved biosynthetic pathways 431:461 conserved biosynthetic pathways 431:461 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 0 24 theme Distinctive 0:10 arg1 Pathways 32:39 Distinctive MS/MS Fragmentation Pathways 0:39 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions 0:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 4 25 gly glycopeptide 656:667 arg2 glycopeptide 656:667 glycopeptide 656:667 glycopeptide 656:667 Such patterns could be used to distinguish between glycopeptide structural isomers. 26663535 2 26 theme isomers 363:369 arg1 identification 338:351 identification 338:351 identification of glycan isomers 338:369 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 2 27 theme conserved 431:439 arg1 pathways 454:461 conserved biosynthetic pathways 431:461 conserved biosynthetic pathways 431:461 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 7 28 theme ion 1017:1019 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 28 theme ion 1017:1019 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 28 theme ion 1017:1019 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 29 theme structures 1159:1168 arg1 number 1129:1134 a number 1127:1134 a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1127:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 30 theme decomposition 1021:1033 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 30 theme decomposition 1021:1033 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 30 theme decomposition 1021:1033 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 2 31 theme glycan 356:361 arg1 isomers 363:369 glycan isomers 356:369 glycan isomers 356:369 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 3 32 theme different 520:528 arg1 patterns 563:570 characteristically different HexNAc oxonium ion fragmentation patterns 501:570 characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure 501:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 0 33 theme Fragmentation 18:30 arg1 Pathways 32:39 Distinctive MS/MS Fragmentation Pathways 0:39 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions 0:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 1 34 gly glycosylation 141:153 arg1 proteins 158:165 proteins 158:165 proteins 158:165 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 6 35 theme abundant 958:965 arg1 ions 987:990 abundant Hex-derived oxonium ions 958:990 abundant Hex-derived oxonium ions 958:990 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 1 36 theme Post-translational 122:139 arg1 glycosylation 141:153 Post-translational glycosylation 122:153 Post-translational glycosylation of proteins 122:165 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 7 37 theme relative 1054:1061 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 37 theme relative 1054:1061 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 37 theme relative 1054:1061 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 3 38 theme HexNAc 530:535 arg1 patterns 563:570 characteristically different HexNAc oxonium ion fragmentation patterns 501:570 characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure 501:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 0 39 theme Glycopeptide-Generated 44:65 arg1 Ions 75:78 Glycopeptide-Generated Oxonium Ions 44:78 Glycopeptide-Generated Oxonium Ions 44:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 7 40 theme well-defined 1139:1150 arg1 structures 1159:1168 well-defined glycan structures 1139:1168 well-defined glycan structures 1139:1168 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 1 41 theme site-specific 212:224 arg1 critical 267:274 critical 267:274 critical 267:274 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 1 41 theme site-specific 212:224 arg1 characterisation 226:241 the site-specific characterisation 208:241 the site-specific characterisation of glycan structures 208:262 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 1 42 theme glycan 246:251 arg1 structures 253:262 glycan structures 246:262 glycan structures 246:262 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 6 43 gly N-glycopeptides 932:946 arg2 N-glycopeptides 932:946 high-mannose N-glycopeptides 919:946 high-mannose N-glycopeptides 919:946 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 7 44 theme glycan 1152:1157 arg1 structures 1159:1168 well-defined glycan structures 1139:1168 well-defined glycan structures 1139:1168 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 45 theme collision 1106:1114 arg1 energy 1116:1121 collision energy 1106:1121 collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1106:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 1 46 theme structures 253:262 arg1 critical 267:274 critical 267:274 critical 267:274 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 1 46 theme structures 253:262 arg1 characterisation 226:241 the site-specific characterisation 208:241 the site-specific characterisation of glycan structures 208:262 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 0 47 theme Ions 75:78 arg1 Pathways 32:39 Distinctive MS/MS Fragmentation Pathways 0:39 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions 0:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 6 48 theme high-mannose 919:930 arg1 N-glycopeptides 932:946 high-mannose N-glycopeptides 919:946 high-mannose N-glycopeptides 919:946 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 7 49 theme energy 1116:1121 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 49 theme energy 1116:1121 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 49 theme energy 1116:1121 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 3 50 theme fragmentation 549:561 arg1 patterns 563:570 characteristically different HexNAc oxonium ion fragmentation patterns 501:570 characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure 501:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 0 51 theme Oxonium 67:73 arg1 Ions 75:78 Glycopeptide-Generated Oxonium Ions 44:78 Glycopeptide-Generated Oxonium Ions 44:78 Distinctive MS/MS Fragmentation Pathways of Glycopeptide-Generated Oxonium Ions Provide Evidence of the Glycan Structure. 26663535 4 52 theme Such 605:608 arg1 patterns 610:617 Such patterns 605:617 Such patterns 605:617 Such patterns could be used to distinguish between glycopeptide structural isomers. 26663535 6 53 theme oxonium 833:839 arg1 ions 841:844 the HexNAc-derived m/z 126 and 144 oxonium ions 798:844 ions 841:844 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 1 54 theme proteins 158:165 arg1 glycosylation 141:153 Post-translational glycosylation 122:153 Post-translational glycosylation of proteins 122:165 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 7 55 theme oxonium 1009:1015 arg1 mechanisms 1035:1044 the oxonium ion decomposition mechanisms 1005:1044 the oxonium ion decomposition mechanisms 1005:1044 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 55 theme oxonium 1009:1015 arg1 abundance 1063:1071 the relative abundance 1050:1071 the relative abundance of oxonium ions 1050:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 55 theme oxonium 1009:1015 arg1 function 1094:1101 a function 1092:1101 a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies 1092:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 7 56 theme glycoproteomic 1218:1231 arg1 studies 1233:1239 future glycoproteomic studies 1211:1239 future glycoproteomic studies 1211:1239 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 2 57 theme glycoproteomic 372:385 arg1 studies 387:393 glycoproteomic studies 372:393 glycoproteomic studies 372:393 Given the challenges regarding identification of glycan isomers, glycoproteomic studies generally rely on the assumption of conserved biosynthetic pathways. 26663535 3 58 theme oxonium 537:543 arg1 patterns 563:570 characteristically different HexNAc oxonium ion fragmentation patterns 501:570 characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure 501:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26663535 6 59 theme different 889:897 arg1 structures 899:908 completely different structures 878:908 completely different structures 878:908 We found that the HexNAc-derived m/z 126 and 144 oxonium ions, differing in mass by H2 O, had completely different structures and that high-mannose N-glycopeptides generated abundant Hex-derived oxonium ions. 26663535 1 60 theme key 172:174 arg1 roles 176:180 key roles 172:180 key roles 172:180 Post-translational glycosylation of proteins play key roles in cellular processes and the site-specific characterisation of glycan structures is critical to understanding these events. 26663535 7 61 theme oxonium 1076:1082 arg1 ions 1084:1087 oxonium ions 1076:1087 oxonium ions 1076:1087 We describe the oxonium ion decomposition mechanisms and the relative abundance of oxonium ions as a function of collision energy for a number of well-defined glycan structures, which provide important information for future glycoproteomic studies. 26663535 3 62 theme glycan 587:592 arg1 structure 594:602 glycan structure 587:602 glycan structure 587:602 However, in a recent study, we found characteristically different HexNAc oxonium ion fragmentation patterns that depend on glycan structure. 26295743 3 0 theme plant 385:389 arg1 biologists 391:400 plant biologists 385:400 plant biologists 385:400 Synthetic chemistry is key to the procurement of molecular probes for plant biologists. 26295743 4 1 theme oligosaccharides 452:467 arg1 assembly 437:444 the automated glycan assembly 416:444 the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks 416:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 1 theme oligosaccharides 452:467 arg1 Described 403:411 Described 403:411 Described 403:411 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 2 theme monosaccharide 479:492 arg1 blocks 503:508 four monosaccharide building blocks 474:508 four monosaccharide building blocks 474:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 3 3 theme Synthetic 315:323 arg1 chemistry 325:333 Synthetic chemistry 315:333 Synthetic chemistry 315:333 Synthetic chemistry is key to the procurement of molecular probes for plant biologists. 26295743 1 4 theme Arabinogalactan 83:97 arg1 proteins 99:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 1 4 theme Arabinogalactan 83:97 arg1 proteoglycans 133:145 heavily glycosylated proteoglycans 112:145 heavily glycosylated proteoglycans in plants 112:155 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 5 5 theme natural 582:588 arg1 arabinogalactans 598:613 natural type-II arabinogalactans 582:613 natural type-II arabinogalactans 582:613 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 1 6 gly glycosylated 120:131 arg1 proteins 99:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 1 6 gly glycosylated 120:131 arg1 proteoglycans 133:145 heavily glycosylated proteoglycans 112:145 heavily glycosylated proteoglycans in plants 112:155 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 5 7 theme type-II 590:596 arg1 arabinogalactans 598:613 natural type-II arabinogalactans 582:613 natural type-II arabinogalactans 582:613 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 0 8 theme Glycan 10:15 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Oligosaccharides 0:44 Automated Glycan Assembly of Oligosaccharides Related to Arabinogalactan Proteins. 26295743 2 9 theme glycan 164:169 arg1 portion 171:177 Their glycan portion 158:177 Their glycan portion 158:177 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26295743 4 10 theme glycan 430:435 arg1 assembly 437:444 the automated glycan assembly 416:444 the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks 416:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 10 theme glycan 430:435 arg1 Described 403:411 Described 403:411 Described 403:411 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 5 11 theme arabinogalactans 598:613 arg1 features 570:577 key structural features 555:577 key structural features of natural type-II arabinogalactans 555:613 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 0 12 theme Automated 0:8 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Oligosaccharides 0:44 Automated Glycan Assembly of Oligosaccharides Related to Arabinogalactan Proteins. 26295743 1 13 theme glycosylated 120:131 arg1 proteins 99:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins 83:106 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 1 13 theme glycosylated 120:131 arg1 proteoglycans 133:145 heavily glycosylated proteoglycans 112:145 heavily glycosylated proteoglycans in plants 112:155 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 5 14 theme key 555:557 arg1 features 570:577 key structural features 555:577 key structural features of natural type-II arabinogalactans 555:613 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 4 15 from assembly 437:444 arg1 blocks 503:508 four monosaccharide building blocks 474:508 four monosaccharide building blocks 474:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 5 16 theme arabinogalactan 643:657 arg1 biology 659:665 arabinogalactan biology 643:665 arabinogalactan biology 643:665 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 5 17 theme structural 559:568 arg1 features 570:577 key structural features 555:577 key structural features of natural type-II arabinogalactans 555:613 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 2 18 theme protein 297:303 arg1 function 305:312 the arabinogalactan protein function 277:312 the arabinogalactan protein function 277:312 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26295743 0 19 theme Oligosaccharides 29:44 arg1 Assembly 17:24 Automated Glycan Assembly 0:24 Automated Glycan Assembly of Oligosaccharides 0:44 Automated Glycan Assembly of Oligosaccharides Related to Arabinogalactan Proteins. 26295743 2 20 theme arabinogalactan 199:213 arg1 polysaccharides 215:229 type-II arabinogalactan polysaccharides 191:229 type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function 191:312 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26295743 2 21 theme arabinogalactan 281:295 arg1 function 305:312 the arabinogalactan protein function 277:312 the arabinogalactan protein function 277:312 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26295743 0 22 theme Arabinogalactan 57:71 arg1 Proteins 73:80 Arabinogalactan Proteins 57:80 Arabinogalactan Proteins 57:80 Automated Glycan Assembly of Oligosaccharides Related to Arabinogalactan Proteins. 26295743 3 23 theme molecular 364:372 arg1 probes 374:379 molecular probes 364:379 molecular probes for plant biologists 364:400 Synthetic chemistry is key to the procurement of molecular probes for plant biologists. 26295743 2 24 theme type-II 191:197 arg1 polysaccharides 215:229 type-II arabinogalactan polysaccharides 191:229 type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function 191:312 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26295743 3 25 theme probes 374:379 arg1 procurement 349:359 the procurement 345:359 the procurement of molecular probes for plant biologists 345:400 Synthetic chemistry is key to the procurement of molecular probes for plant biologists. 26295743 4 26 theme automated 420:428 arg1 assembly 437:444 the automated glycan assembly 416:444 the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks 416:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 26 theme automated 420:428 arg1 Described 403:411 Described 403:411 Described 403:411 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 5 27 theme linear 517:522 arg1 tools 633:637 tools 633:637 tools for arabinogalactan biology 633:665 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 5 27 theme linear 517:522 arg1 glycans 537:543 These linear and branched glycans 511:543 These linear and branched glycans 511:543 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 4 28 from blocks 503:508 arg1 Described 403:411 Described 403:411 Described 403:411 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 28 from blocks 503:508 arg1 assembly 437:444 the automated glycan assembly 416:444 the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks 416:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 28 from blocks 503:508 arg1 oligosaccharides 452:467 14 oligosaccharides 449:467 14 oligosaccharides from four monosaccharide building blocks 449:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 4 29 theme building 494:501 arg1 blocks 503:508 four monosaccharide building blocks 474:508 four monosaccharide building blocks 474:508 Described is the automated glycan assembly of 14 oligosaccharides from four monosaccharide building blocks. 26295743 1 30 from proteoglycans 133:145 arg1 plants 150:155 plants 150:155 plants 150:155 Arabinogalactan proteins are heavily glycosylated proteoglycans in plants. 26295743 5 31 theme branched 528:535 arg1 tools 633:637 tools 633:637 tools for arabinogalactan biology 633:665 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 5 31 theme branched 528:535 arg1 glycans 537:543 These linear and branched glycans 511:543 These linear and branched glycans 511:543 These linear and branched glycans represent key structural features of natural type-II arabinogalactans and will serve as tools for arabinogalactan biology. 26295743 2 32 theme function 305:312 arg1 assignment 263:272 the assignment 259:272 the assignment of the arabinogalactan protein function 259:312 Their glycan portion consists of type-II arabinogalactan polysaccharides whose heterogeneity hampers the assignment of the arabinogalactan protein function. 26912703 0 0 theme A 82:82 arg1 glycosylation 84:96 lipid A glycosylation 76:96 lipid A glycosylation 76:96 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 0 1 theme aminoarabinose 14:27 arg1 ArnT 41:44 aminoarabinose transferase ArnT 14:44 aminoarabinose transferase ArnT 14:44 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 0 2 theme lipid 76:80 arg1 glycosylation 84:96 lipid A glycosylation 76:96 lipid A glycosylation 76:96 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 2 3 theme lipid 388:392 arg1 A 394:394 lipid A 388:394 lipid A 388:394 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 3 theme lipid 388:392 arg1 reaction 399:406 a reaction 397:406 a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 397:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 6 4 theme mutagenesis 960:970 arg1 experiments 972:982 Functional mutagenesis experiments 949:982 Functional mutagenesis experiments based on these structures 949:1008 Functional mutagenesis experiments based on these structures suggest a mechanistic model for ArnT family enzymes. 26912703 3 5 theme lipid 631:635 arg1 phosphate 658:666 the lipid carrier undecaprenyl phosphate 627:666 the lipid carrier undecaprenyl phosphate 627:666 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 6 6 theme Functional 949:958 arg1 experiments 972:982 Functional mutagenesis experiments 949:982 Functional mutagenesis experiments based on these structures 949:1008 Functional mutagenesis experiments based on these structures suggest a mechanistic model for ArnT family enzymes. 26912703 2 7 theme membrane 306:313 arg1 modification 270:281 charge modification 263:281 charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 263:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 6 8 theme family 1047:1052 arg1 enzymes 1054:1060 ArnT family enzymes 1042:1060 ArnT family enzymes 1042:1060 Functional mutagenesis experiments based on these structures suggest a mechanistic model for ArnT family enzymes. 26912703 3 9 from resolution 693:702 arg1 phosphate 658:666 the lipid carrier undecaprenyl phosphate 627:666 the lipid carrier undecaprenyl phosphate 627:666 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 2 10 theme lipid-to-lipid 443:456 arg1 glycosyltransferase 458:476 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 11 theme outer 300:304 arg1 membrane 306:313 the bacterial outer membrane 286:313 the bacterial outer membrane 286:313 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 12 theme bacterial 290:298 arg1 membrane 306:313 the bacterial outer membrane 286:313 the bacterial outer membrane 286:313 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 5 13 theme phosphate 862:870 arg1 binding 872:878 undecaprenyl phosphate binding 849:878 undecaprenyl phosphate binding 849:878 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 5 14 theme structural 814:823 arg1 rearrangement 825:837 A structural rearrangement 812:837 A structural rearrangement 812:837 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 1 15 theme multidrug-resistant 169:187 arg1 infections 189:198 multidrug-resistant infections 169:198 multidrug-resistant infections 169:198 Polymyxins are antibiotics used in the last line of defense to combat multidrug-resistant infections by Gram-negative bacteria. 26912703 3 16 with complex 614:620 arg1 phosphate 658:666 the lipid carrier undecaprenyl phosphate 627:666 the lipid carrier undecaprenyl phosphate 627:666 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 6 17 theme ArnT 1042:1045 arg1 enzymes 1054:1060 ArnT family enzymes 1042:1060 ArnT family enzymes 1042:1060 Functional mutagenesis experiments based on these structures suggest a mechanistic model for ArnT family enzymes. 26912703 6 18 theme mechanistic 1020:1030 arg1 model 1032:1036 a mechanistic model 1018:1036 a mechanistic model for ArnT family enzymes 1018:1060 Functional mutagenesis experiments based on these structures suggest a mechanistic model for ArnT family enzymes. 26912703 5 19 theme lipid 932:936 arg1 binding 940:946 lipid A binding 932:946 lipid A binding 932:946 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 0 20 theme transferase 29:39 arg1 ArnT 41:44 aminoarabinose transferase ArnT 14:44 aminoarabinose transferase ArnT 14:44 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 3 21 theme carrier 637:643 arg1 phosphate 658:666 the lipid carrier undecaprenyl phosphate 627:666 the lipid carrier undecaprenyl phosphate 627:666 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 4 22 theme active 799:804 arg1 site 806:809 the active site 795:809 the active site 795:809 The structures show cavities for both lipidic substrates, which converge at the active site. 26912703 2 23 theme 4-amino-4-deoxy-L-arabinose 478:504 arg1 ArnT 519:522 ArnT 519:522 ArnT 519:522 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 23 theme 4-amino-4-deoxy-L-arabinose 478:504 arg1 transferase 506:516 4-amino-4-deoxy-L-arabinose transferase 478:516 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 1 24 theme defense 151:157 arg1 line 143:146 the last line 134:146 the last line of defense 134:157 Polymyxins are antibiotics used in the last line of defense to combat multidrug-resistant infections by Gram-negative bacteria. 26912703 1 25 theme Gram-negative 203:215 arg1 bacteria 217:224 Gram-negative bacteria 203:224 Gram-negative bacteria 203:224 Polymyxins are antibiotics used in the last line of defense to combat multidrug-resistant infections by Gram-negative bacteria. 26912703 2 26 attach attachment 324:333 arg1 A 394:394 lipid A 388:394 lipid A 388:394 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 26 attach attachment 324:333 arg1 reaction 399:406 a reaction 397:406 a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 397:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 26 attach attachment 324:333 arg2 4-amino-4-deoxy-l-arabinose 357:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 5 27 theme active 902:907 arg1 site 909:912 the active site 898:912 the active site 898:912 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 3 28 theme undecaprenyl 645:656 arg1 phosphate 658:666 the lipid carrier undecaprenyl phosphate 627:666 the lipid carrier undecaprenyl phosphate 627:666 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 2 29 theme membrane 434:441 arg1 glycosyltransferase 458:476 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 0 30 theme ArnT 41:44 arg1 Structures 0:9 Structures 0:9 Structures of aminoarabinose transferase ArnT 0:44 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 2 31 with modification 270:281 arg1 attachment 324:333 the attachment 320:333 the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 320:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 32 theme integral 425:432 arg1 glycosyltransferase 458:476 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 0 33 theme molecular 56:64 arg1 basis 66:70 a molecular basis 54:70 a molecular basis for lipid A glycosylation 54:96 Structures of aminoarabinose transferase ArnT suggest a molecular basis for lipid A glycosylation. 26912703 5 34 theme A 938:938 arg1 binding 940:946 lipid A binding 932:946 lipid A binding 932:946 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 3 35 theme angstrom 684:691 arg1 resolution 693:702 2.8 and 3.2 angstrom resolution 672:702 2.8 and 3.2 angstrom resolution 672:702 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 2 36 dep glycosyltransferase 458:476 arg1 ArnT 519:522 ArnT 519:522 ArnT 519:522 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 36 dep glycosyltransferase 458:476 arg1 transferase 506:516 4-amino-4-deoxy-L-arabinose transferase 478:516 the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 421:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 3 37 theme crystal 542:548 arg1 structures 550:559 crystal structures 542:559 crystal structures of ArnT from Cupriavidus metallidurans 542:598 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 5 38 theme undecaprenyl 849:860 arg1 phosphate 862:870 undecaprenyl phosphate 849:870 undecaprenyl phosphate binding 849:878 A structural rearrangement occurs on undecaprenyl phosphate binding, which stabilizes the active site and likely allows lipid A binding. 26912703 2 39 theme Polymyxin 227:235 arg1 resistance 237:246 Polymyxin resistance 227:246 Polymyxin resistance 227:246 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 3 40 theme ArnT 564:567 arg1 structures 550:559 crystal structures 542:559 crystal structures of ArnT from Cupriavidus metallidurans 542:598 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 2 41 theme 4-amino-4-deoxy-l-arabinose 357:383 arg1 attachment 324:333 the attachment 320:333 the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 320:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 42 theme sugar 351:355 arg1 4-amino-4-deoxy-l-arabinose 357:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 43 theme cationic 342:349 arg1 4-amino-4-deoxy-l-arabinose 357:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 the cationic sugar 4-amino-4-deoxy-l-arabinose 338:383 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 44 mod modification 270:281 arg1 membrane 306:313 the bacterial outer membrane 286:313 the bacterial outer membrane 286:313 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 2 44 mod modification 270:281 arg3 charge 263:268 charge modification 263:281 charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 263:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 1 45 theme last 138:141 arg1 line 143:146 the last line 134:146 the last line of defense 134:157 Polymyxins are antibiotics used in the last line of defense to combat multidrug-resistant infections by Gram-negative bacteria. 26912703 2 46 theme charge 263:268 arg1 modification 270:281 charge modification 263:281 charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT) 263:523 Polymyxin resistance arises through charge modification of the bacterial outer membrane with the attachment of the cationic sugar 4-amino-4-deoxy-l-arabinose to lipid A, a reaction catalyzed by the integral membrane lipid-to-lipid glycosyltransferase 4-amino-4-deoxy-L-arabinose transferase (ArnT). 26912703 3 47 theme Cupriavidus 574:584 arg1 metallidurans 586:598 Cupriavidus metallidurans 574:598 Cupriavidus metallidurans 574:598 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 3 48 from metallidurans 586:598 arg1 structures 550:559 crystal structures 542:559 crystal structures of ArnT from Cupriavidus metallidurans 542:598 Here, we report crystal structures of ArnT from Cupriavidus metallidurans, alone and in complex with the lipid carrier undecaprenyl phosphate, at 2.8 and 3.2 angstrom resolution, respectively. 26912703 4 49 theme lipidic 757:763 arg1 substrates 765:774 both lipidic substrates 752:774 both lipidic substrates 752:774 The structures show cavities for both lipidic substrates, which converge at the active site. 25268221 0 0 theme ion 89:91 arg1 spectrometry 107:118 traveling wave ion mobility-mass spectrometry 74:118 traveling wave ion mobility-mass spectrometry 74:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 9 1 used used 1617:1620 arg2 CCSs 1574:1577 reference CCSs 1564:1577 reference CCSs acquired in a different drift gas 1564:1611 Our data also indicate that a considerably increased error has to be taken into account when reference CCSs acquired in a different drift gas are used for calibration. 25268221 8 2 theme high 1381:1384 arg1 chromatography 1405:1418 high performance liquid chromatography 1381:1418 high performance liquid chromatography 1381:1418 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 1 3 theme post-translational 161:178 arg1 modifications 180:192 the most common post-translational modifications 145:192 the most common post-translational modifications occurring in proteins 145:214 Glycosylation is one of the most common post-translational modifications occurring in proteins. 25268221 8 4 theme performance 1386:1396 arg1 chromatography 1405:1418 high performance liquid chromatography 1381:1418 high performance liquid chromatography 1381:1418 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 0 5 theme wave 84:87 arg1 spectrometry 107:118 traveling wave ion mobility-mass spectrometry 74:118 traveling wave ion mobility-mass spectrometry 74:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 5 6 theme wave 1044:1047 arg1 field 1049:1053 a nonuniform traveling wave field 1021:1053 a nonuniform traveling wave field 1021:1053 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 1 7 theme modifications 180:192 arg1 one 138:140 one 138:140 one 138:140 Glycosylation is one of the most common post-translational modifications occurring in proteins. 25268221 1 7 theme modifications 180:192 arg1 modifications 180:192 the most common post-translational modifications 145:192 the most common post-translational modifications occurring in proteins 145:214 Glycosylation is one of the most common post-translational modifications occurring in proteins. 25268221 4 8 used used 882:885 arg2 criteria 908:915 additional search criteria 890:915 additional search criteria for structural identification 890:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 8 used used 882:885 arg2 CCSs 820:823 CCSs 820:823 CCSs 820:823 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 8 used used 882:885 arg2 sections 810:817 collision cross sections 794:817 collision cross sections (CCSs) 794:824 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 9 theme structural 921:930 arg1 identification 932:945 structural identification 921:945 structural identification 921:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 8 10 theme CCS 1454:1456 arg1 estimations 1458:1468 CCS estimations 1454:1468 CCS estimations 1454:1468 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 9 11 theme different 1593:1601 arg1 gas 1609:1611 a different drift gas 1591:1611 a different drift gas 1591:1611 Our data also indicate that a considerably increased error has to be taken into account when reference CCSs acquired in a different drift gas are used for calibration. 25268221 3 12 theme carbohydrates 684:696 arg1 technique 620:628 a promising technique 608:628 a promising technique for the separation 608:647 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 3 12 theme carbohydrates 684:696 arg1 analysis 664:671 structural analysis 653:671 structural analysis of complex carbohydrates 653:696 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 0 13 theme mobility-mass 93:105 arg1 spectrometry 107:118 traveling wave ion mobility-mass spectrometry 74:118 traveling wave ion mobility-mass spectrometry 74:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 9 14 theme drift 1603:1607 arg1 gas 1609:1611 a different drift gas 1591:1611 a different drift gas 1591:1611 Our data also indicate that a considerably increased error has to be taken into account when reference CCSs acquired in a different drift gas are used for calibration. 25268221 7 15 theme data 1210:1213 arg1 set 1215:1217 a calibration data set 1196:1217 a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments 1196:1308 Here, we present a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments. 25268221 4 16 theme drift 760:764 arg1 times 766:770 the measured drift times 747:770 the measured drift times 747:770 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 17 theme growing 704:710 arg1 interest 712:719 This growing interest 699:719 This growing interest 699:719 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 3 18 theme mobility-mass 473:485 arg1 IM-MS 501:505 IM-MS 501:505 IM-MS 501:505 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 3 18 theme mobility-mass 473:485 arg1 spectrometry 487:498 ion mobility-mass spectrometry 469:498 ion mobility-mass spectrometry (IM-MS) 469:506 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 3 18 theme mobility-mass 473:485 arg1 technique 511:519 a technique 509:519 a technique in which ions are separated according to their mass, charge, and shape 509:590 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 5 19 theme IM 1086:1087 arg1 cell 1089:1092 the IM cell 1082:1092 the IM cell 1082:1092 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 6 20 theme CCS 1108:1110 arg1 measurements 1112:1123 CCS measurements 1108:1123 CCS measurements 1108:1123 As a result, CCS measurements cannot be performed directly and require calibration. 25268221 6 20 theme CCS 1108:1110 arg1 calibration 1166:1176 calibration 1166:1176 calibration 1166:1176 As a result, CCS measurements cannot be performed directly and require calibration. 25268221 6 20 theme CCS 1108:1110 arg1 result 1100:1105 a result 1098:1105 a result 1098:1105 As a result, CCS measurements cannot be performed directly and require calibration. 25268221 2 21 theme monosaccharide 421:434 arg1 composition 436:446 an identical monosaccharide composition 408:446 an identical monosaccharide composition 408:446 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 0 22 theme collision 11:19 arg1 sections 27:34 collision cross sections 11:34 collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry 11:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 2 23 theme modern 339:344 arg1 glycoproteomics 346:360 modern glycoproteomics 339:360 modern glycoproteomics 339:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 24 theme identical 411:419 arg1 composition 436:446 an identical monosaccharide composition 408:446 an identical monosaccharide composition 408:446 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 7 25 theme calibration 1198:1208 arg1 set 1215:1217 a calibration data set 1196:1217 a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments 1196:1308 Here, we present a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments. 25268221 5 26 theme nonuniform 1023:1032 arg1 field 1049:1053 a nonuniform traveling wave field 1021:1053 a nonuniform traveling wave field 1021:1053 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 3 27 theme ion 469:471 arg1 IM-MS 501:505 IM-MS 501:505 IM-MS 501:505 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 3 27 theme ion 469:471 arg1 spectrometry 487:498 ion mobility-mass spectrometry 469:498 ion mobility-mass spectrometry (IM-MS) 469:506 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 3 27 theme ion 469:471 arg1 technique 511:519 a technique 509:519 a technique in which ions are separated according to their mass, charge, and shape 509:590 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 8 28 theme suitable 1431:1438 arg1 calibrant 1440:1448 a suitable calibrant 1429:1448 a suitable calibrant for CCS estimations 1429:1468 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 8 28 theme suitable 1431:1438 arg1 dextran 1334:1340 dextran 1334:1340 dextran 1334:1340 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 5 29 theme used 979:982 arg1 instruments 1001:1011 the currently used commercial IM-MS instruments 965:1011 the currently used commercial IM-MS instruments 965:1011 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 4 30 theme additional 890:899 arg1 sections 810:817 collision cross sections 794:817 collision cross sections (CCSs) 794:824 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 30 theme additional 890:899 arg1 criteria 908:915 additional search criteria 890:915 additional search criteria for structural identification 890:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 2 31 theme challenges 325:334 arg1 challenges 325:334 the greatest challenges 312:334 the greatest challenges in modern glycoproteomics 312:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 31 theme challenges 325:334 arg1 one 305:307 one 305:307 one 305:307 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 31 theme challenges 325:334 arg1 characterization 239:254 A detailed structural characterization 217:254 A detailed structural characterization of the involved carbohydrates 217:284 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 5 32 theme IM-MS 995:999 arg1 instruments 1001:1011 the currently used commercial IM-MS instruments 965:1011 the currently used commercial IM-MS instruments 965:1011 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 0 33 theme cross 21:25 arg1 sections 27:34 collision cross sections 11:34 collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry 11:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 8 34 theme liquid 1398:1403 arg1 chromatography 1405:1418 high performance liquid chromatography 1381:1418 high performance liquid chromatography 1381:1418 Moreover, we show that dextran, already widely used as a calibrant in high performance liquid chromatography, is also a suitable calibrant for CCS estimations. 25268221 2 35 theme greatest 316:323 arg1 challenges 325:334 the greatest challenges 312:334 the greatest challenges in modern glycoproteomics 312:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 7 36 theme charged 1272:1278 arg1 N-glycans 1280:1288 negatively charged N-glycans 1261:1288 negatively charged N-glycans 1261:1288 Here, we present a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments. 25268221 5 37 theme commercial 984:993 arg1 instruments 1001:1011 the currently used commercial IM-MS instruments 965:1011 the currently used commercial IM-MS instruments 965:1011 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 3 38 theme structural 653:662 arg1 analysis 664:671 structural analysis 653:671 structural analysis of complex carbohydrates 653:696 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 2 39 theme carbohydrates 272:284 arg1 challenges 325:334 the greatest challenges 312:334 the greatest challenges in modern glycoproteomics 312:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 39 theme carbohydrates 272:284 arg1 one 305:307 one 305:307 one 305:307 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 39 theme carbohydrates 272:284 arg1 characterization 239:254 A detailed structural characterization 217:254 A detailed structural characterization of the involved carbohydrates 217:284 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 4 40 theme search 901:906 arg1 sections 810:817 collision cross sections 794:817 collision cross sections (CCSs) 794:824 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 40 theme search 901:906 arg1 criteria 908:915 additional search criteria 890:915 additional search criteria for structural identification 890:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 2 41 theme involved 263:270 arg1 carbohydrates 272:284 the involved carbohydrates 259:284 the involved carbohydrates 259:284 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 0 42 theme N-glycans 58:66 arg1 sections 27:34 collision cross sections 11:34 collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry 11:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 9 43 theme increased 1514:1522 arg1 error 1524:1528 a considerably increased error 1499:1528 a considerably increased error 1499:1528 Our data also indicate that a considerably increased error has to be taken into account when reference CCSs acquired in a different drift gas are used for calibration. 25268221 5 44 theme traveling 1034:1042 arg1 field 1049:1053 a nonuniform traveling wave field 1021:1053 a nonuniform traveling wave field 1021:1053 However, most of the currently used commercial IM-MS instruments utilize a nonuniform traveling wave field to propel the ions through the IM cell. 25268221 2 45 theme multiple 369:376 arg1 regio- 378:383 multiple regio- 369:383 multiple regio- 369:383 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 0 46 theme charged 50:56 arg1 N-glycans 58:66 negatively charged N-glycans 39:66 negatively charged N-glycans using traveling wave ion mobility-mass spectrometry 39:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 3 47 theme complex 676:682 arg1 carbohydrates 684:696 complex carbohydrates 676:696 complex carbohydrates 676:696 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 2 48 with regio- 378:383 arg1 composition 436:446 an identical monosaccharide composition 408:446 an identical monosaccharide composition 408:446 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 7 49 theme reference 1242:1250 arg1 CCSs 1252:1255 over 500 reference CCSs 1233:1255 over 500 reference CCSs for negatively charged N-glycans and their fragments 1233:1308 Here, we present a calibration data set consisting of over 500 reference CCSs for negatively charged N-glycans and their fragments. 25268221 4 50 theme collision 794:802 arg1 CCSs 820:823 CCSs 820:823 CCSs 820:823 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 50 theme collision 794:802 arg1 sections 810:817 collision cross sections 794:817 collision cross sections (CCSs) 794:824 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 50 theme collision 794:802 arg1 criteria 908:915 additional search criteria 890:915 additional search criteria for structural identification 890:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 2 51 theme structural 228:237 arg1 challenges 325:334 the greatest challenges 312:334 the greatest challenges in modern glycoproteomics 312:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 51 theme structural 228:237 arg1 one 305:307 one 305:307 one 305:307 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 51 theme structural 228:237 arg1 characterization 239:254 A detailed structural characterization 217:254 A detailed structural characterization of the involved carbohydrates 217:284 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 4 52 theme measured 751:758 arg1 times 766:770 the measured drift times 747:770 the measured drift times 747:770 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 2 53 theme detailed 219:226 arg1 challenges 325:334 the greatest challenges 312:334 the greatest challenges in modern glycoproteomics 312:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 53 theme detailed 219:226 arg1 one 305:307 one 305:307 one 305:307 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 2 53 theme detailed 219:226 arg1 characterization 239:254 A detailed structural characterization 217:254 A detailed structural characterization of the involved carbohydrates 217:284 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 9 54 theme reference 1564:1572 arg1 CCSs 1574:1577 reference CCSs 1564:1577 reference CCSs acquired in a different drift gas 1564:1611 Our data also indicate that a considerably increased error has to be taken into account when reference CCSs acquired in a different drift gas are used for calibration. 25268221 0 55 theme traveling 74:82 arg1 spectrometry 107:118 traveling wave ion mobility-mass spectrometry 74:118 traveling wave ion mobility-mass spectrometry 74:118 Estimating collision cross sections of negatively charged N-glycans using traveling wave ion mobility-mass spectrometry. 25268221 2 56 with stereoisomers 389:401 arg1 composition 436:446 an identical monosaccharide composition 408:446 an identical monosaccharide composition 408:446 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 25268221 4 57 theme cross 804:808 arg1 CCSs 820:823 CCSs 820:823 CCSs 820:823 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 57 theme cross 804:808 arg1 sections 810:817 collision cross sections 794:817 collision cross sections (CCSs) 794:824 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 4 57 theme cross 804:808 arg1 criteria 908:915 additional search criteria 890:915 additional search criteria for structural identification 890:945 This growing interest is based on the fact that the measured drift times can be converted into collision cross sections (CCSs), which can be compared, implemented into databases, and used as additional search criteria for structural identification. 25268221 3 58 theme promising 610:618 arg1 technique 620:628 a promising technique 608:628 a promising technique for the separation 608:647 Recently, ion mobility-mass spectrometry (IM-MS), a technique in which ions are separated according to their mass, charge, and shape, has evolved as a promising technique for the separation and structural analysis of complex carbohydrates. 25268221 1 59 theme common 154:159 arg1 modifications 180:192 the most common post-translational modifications 145:192 the most common post-translational modifications occurring in proteins 145:214 Glycosylation is one of the most common post-translational modifications occurring in proteins. 25268221 2 60 from challenges 325:334 arg1 glycoproteomics 346:360 modern glycoproteomics 339:360 modern glycoproteomics 339:360 A detailed structural characterization of the involved carbohydrates, however, is still one of the greatest challenges in modern glycoproteomics, since multiple regio- and stereoisomers with an identical monosaccharide composition may exist. 27295180 5 0 theme validated 664:672 arg1 biomarker 674:682 no validated biomarker 661:682 no validated biomarker for predicting treatment outcomes in therapeutic settings 661:740 In particular, there has been no validated biomarker for predicting treatment outcomes in therapeutic settings. 27295180 7 1 theme various 1043:1049 arg1 cancers 1051:1057 various cancers 1043:1057 various cancers 1043:1057 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 7 2 theme glycan 920:925 arg1 structures 927:936 The primary glycan structures 908:936 The primary glycan structures of AGP 908:943 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 4 3 theme major 602:606 arg1 challenge 620:628 a major therapeutic challenge 600:628 a major therapeutic challenge 600:628 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 0 4 from Glycans 12:18 arg1 Glycoprotein 31:42 α1-Acid Glycoprotein 23:42 α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients 23:109 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 12 5 theme treatment 1899:1907 arg1 intervention 1909:1920 treatment intervention 1899:1920 treatment intervention 1899:1920 Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention. 27295180 6 6 theme crossed 829:835 arg1 affinoimmunoelectrophoresis 837:863 a crossed affinoimmunoelectrophoresis 827:863 a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody 827:905 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 1 7 theme surgical 171:178 arg1 resection 180:188 surgical resection 171:188 surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy 171:265 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 10 8 theme various 1423:1429 arg1 patients 1438:1445 various cancer patients 1423:1445 various cancer patients 1423:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 0 9 theme Patients 102:109 arg1 Prognosis 82:90 Prognosis 82:90 Prognosis of Cancer Patients 82:109 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 0 9 theme Patients 102:109 arg1 Outcomes 69:76 Monitoring Treatment Outcomes 48:76 Monitoring Treatment Outcomes 48:76 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 10 10 theme FUCAGP 1394:1399 arg1 levels 1384:1389 levels 1384:1389 levels of FUCAGP in serum samples from various cancer patients 1384:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 12 11 theme patients 1826:1833 arg1 following-up 1806:1817 following-up 1806:1817 following-up of the patients after operation 1806:1849 Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention. 27295180 8 12 theme healthy 1214:1220 arg1 controls 1222:1229 the healthy controls 1210:1229 the healthy controls 1210:1229 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 6 13 gly glycoforms 771:780 arg1 AGP 813:815 AGP 813:815 AGP 813:815 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 13 gly glycoforms 771:780 arg1 glycoprotein 799:810 serum α1-acid glycoprotein 785:810 serum α1-acid glycoprotein (AGP) 785:816 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 0 14 theme Cancer 95:100 arg1 Patients 102:109 Cancer Patients 95:109 Cancer Patients 95:109 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 7 15 theme patients 1029:1036 arg1 number 1019:1024 a large number 1011:1024 a large number of patients with various cancers 1011:1057 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 7 16 theme mass 969:972 arg1 spectrometer 974:985 a mass spectrometer 967:985 a mass spectrometer 967:985 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 9 17 located found 1283:1287 arg2 levels 1261:1266 strikingly elevated levels 1241:1266 strikingly elevated levels of FUCAGP 1241:1276 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 9 17 located found 1283:1287 arg1 patients 1292:1299 patients 1292:1299 patients with poor prognosis 1292:1319 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 4 18 theme therapeutic 608:618 arg1 challenge 620:628 a major therapeutic challenge 600:628 a major therapeutic challenge 600:628 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 9 19 theme poor 1306:1309 arg1 prognosis 1311:1319 poor prognosis 1306:1319 poor prognosis 1306:1319 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 12 20 theme intervention 1909:1920 arg1 biomarker 1886:1894 clinically relevant biomarker 1866:1894 clinically relevant biomarker of treatment intervention 1866:1920 Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention. 27295180 9 21 with patients 1292:1299 arg1 prognosis 1311:1319 poor prognosis 1306:1319 poor prognosis 1306:1319 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 3 22 theme tumor 454:458 arg1 diagnosis 467:475 tumor marker diagnosis 454:475 tumor marker diagnosis 454:475 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 7 23 with patients 1029:1036 arg1 cancers 1051:1057 various cancers 1043:1057 various cancers 1043:1057 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 11 24 theme various 1741:1747 arg1 chemotherapies 1749:1762 various chemotherapies 1741:1762 various chemotherapies 1741:1762 FUCAGP level determined diligently by using a mass spectrometer was found to change along with disease prognosis as well as with responses to treatments, in particular, to various chemotherapies. 27295180 9 25 with patients 1332:1339 arg1 prognosis 1351:1359 good prognosis 1346:1359 good prognosis 1346:1359 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 8 26 theme cancer 1177:1182 arg1 patients 1184:1191 cancer patients 1177:1191 cancer patients 1177:1191 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 8 27 gly α1,3fucosylated 1099:1113 arg1 glycans 1115:1121 α1,3fucosylated glycans 1099:1121 α1,3fucosylated glycans 1099:1121 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 3 28 theme marker 460:465 arg1 diagnosis 467:475 tumor marker diagnosis 454:475 tumor marker diagnosis 454:475 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 3 29 theme tomography 417:426 arg1 surveillance 433:444 computed tomography (CT) surveillance 408:444 computed tomography (CT) surveillance 408:444 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 4 30 theme early 498:502 arg1 evidence 504:511 early evidence 498:511 early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker 498:590 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 8 31 theme relative 1077:1084 arg1 abundance 1086:1094 the relative abundance 1073:1094 the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) 1073:1137 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 8 31 theme relative 1077:1084 arg1 high 1169:1172 high 1169:1172 high 1169:1172 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 10 32 theme serum 1404:1408 arg1 samples 1410:1416 serum samples 1404:1416 serum samples from various cancer patients 1404:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 1 33 theme malignant 193:201 arg1 tumors 203:208 malignant tumors 193:208 malignant tumors following by adjuvant chemotherapy and chemoradiotherapy 193:265 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 0 34 theme Fucosylated 0:10 arg1 Glycans 12:18 Fucosylated Glycans 0:18 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 0:110 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 4 35 theme relevant 573:580 arg1 biomarker 582:590 a clinically relevant biomarker 560:590 a clinically relevant biomarker 560:590 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 10 36 theme cancer 1431:1436 arg1 patients 1438:1445 various cancer patients 1423:1445 various cancer patients 1423:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 9 37 theme elevated 1252:1259 arg1 levels 1261:1266 strikingly elevated levels 1241:1266 strikingly elevated levels of FUCAGP 1241:1276 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 11 38 theme mass 1615:1618 arg1 spectrometer 1620:1631 a mass spectrometer 1613:1631 a mass spectrometer 1613:1631 FUCAGP level determined diligently by using a mass spectrometer was found to change along with disease prognosis as well as with responses to treatments, in particular, to various chemotherapies. 27295180 1 39 theme tumors 203:208 arg1 resection 180:188 surgical resection 171:188 surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy 171:265 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 0 40 theme α1-Acid 23:29 arg1 Glycoprotein 31:42 α1-Acid Glycoprotein 23:42 α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients 23:109 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 6 41 theme glycoprotein 799:810 arg1 glycoforms 771:780 glycoforms 771:780 glycoforms of serum α1-acid glycoprotein (AGP) 771:816 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 4 42 with tumors 548:553 arg1 biomarker 582:590 a clinically relevant biomarker 560:590 a clinically relevant biomarker 560:590 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 3 43 theme computed 408:415 arg1 tomography 417:426 computed tomography 408:426 computed tomography (CT) surveillance 408:444 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 3 43 theme computed 408:415 arg1 CT 429:430 CT 429:430 CT 429:430 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 6 44 gly glycoprotein 799:810 arg1 AGP 813:815 AGP 813:815 AGP 813:815 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 44 gly glycoprotein 799:810 arg1 glycoprotein 799:810 serum α1-acid glycoprotein 785:810 serum α1-acid glycoprotein (AGP) 785:816 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 45 with affinoimmunoelectrophoresis 837:863 arg1 lectins 874:880 two lectins 870:880 two lectins 870:880 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 45 with affinoimmunoelectrophoresis 837:863 arg1 antibody 898:905 an anti-AGP antibody 886:905 an anti-AGP antibody 886:905 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 8 46 theme glycans 1115:1121 arg1 abundance 1086:1094 the relative abundance 1073:1094 the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) 1073:1137 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 8 46 theme glycans 1115:1121 arg1 high 1169:1172 high 1169:1172 high 1169:1172 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 6 47 theme anti-AGP 889:896 arg1 antibody 898:905 an anti-AGP antibody 886:905 an anti-AGP antibody 886:905 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 1 48 theme adjuvant 223:230 arg1 chemotherapy 232:243 adjuvant chemotherapy 223:243 adjuvant chemotherapy 223:243 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 4 49 theme tumors 548:553 arg1 recurrence 516:525 recurrence 516:525 recurrence 516:525 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 8 50 theme α1,3fucosylated 1099:1113 arg1 glycans 1115:1121 α1,3fucosylated glycans 1099:1121 α1,3fucosylated glycans 1099:1121 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 11 51 theme disease 1664:1670 arg1 prognosis 1672:1680 disease prognosis 1664:1680 disease prognosis 1664:1680 FUCAGP level determined diligently by using a mass spectrometer was found to change along with disease prognosis as well as with responses to treatments, in particular, to various chemotherapies. 27295180 4 52 theme metastasis 534:543 arg1 evidence 504:511 early evidence 498:511 early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker 498:590 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 10 53 theme several 1539:1545 arg1 years 1547:1551 several years 1539:1551 several years post operation 1539:1566 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 0 54 theme Treatment 59:67 arg1 Outcomes 69:76 Monitoring Treatment Outcomes 48:76 Monitoring Treatment Outcomes 48:76 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 9 55 theme good 1346:1349 arg1 prognosis 1351:1359 good prognosis 1346:1359 good prognosis 1346:1359 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 5 56 theme treatment 699:707 arg1 outcomes 709:716 treatment outcomes 699:716 treatment outcomes 699:716 In particular, there has been no validated biomarker for predicting treatment outcomes in therapeutic settings. 27295180 6 57 theme serum 785:789 arg1 AGP 813:815 AGP 813:815 AGP 813:815 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 57 theme serum 785:789 arg1 glycoprotein 799:810 serum α1-acid glycoprotein 785:810 serum α1-acid glycoprotein (AGP) 785:816 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 1 58 theme standard 116:123 arg1 option 135:140 One standard treatment option 112:140 One standard treatment option for advanced-stage cancer 112:166 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 0 59 theme Monitoring 48:57 arg1 Outcomes 69:76 Monitoring Treatment Outcomes 48:76 Monitoring Treatment Outcomes 48:76 Fucosylated Glycans in α1-Acid Glycoprotein for Monitoring Treatment Outcomes and Prognosis of Cancer Patients. 27295180 3 60 theme time 346:349 arg1 course 351:356 the time course 342:356 the time course of treatments 342:370 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 8 61 from patients 1184:1191 arg1 abundance 1086:1094 the relative abundance 1073:1094 the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) 1073:1137 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 8 61 from patients 1184:1191 arg1 high 1169:1172 high 1169:1172 high 1169:1172 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 7 62 theme large 1013:1017 arg1 number 1019:1024 a large number 1011:1024 a large number of patients with various cancers 1011:1057 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 1 63 theme treatment 125:133 arg1 option 135:140 One standard treatment option 112:140 One standard treatment option for advanced-stage cancer 112:166 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 7 64 theme AGP 941:943 arg1 structures 927:936 The primary glycan structures 908:936 The primary glycan structures of AGP 908:943 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 5 65 from biomarker 674:682 arg1 particular 634:643 particular 634:643 particular 634:643 In particular, there has been no validated biomarker for predicting treatment outcomes in therapeutic settings. 27295180 8 66 from high 1169:1172 arg1 patients 1184:1191 cancer patients 1177:1191 cancer patients 1177:1191 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 11 67 theme FUCAGP 1569:1574 arg1 level 1576:1580 FUCAGP level 1569:1580 FUCAGP level determined diligently by using a mass spectrometer 1569:1631 FUCAGP level determined diligently by using a mass spectrometer was found to change along with disease prognosis as well as with responses to treatments, in particular, to various chemotherapies. 27295180 10 68 theme current 1369:1375 arg1 study 1377:1381 the current study 1365:1381 the current study 1365:1381 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 12 69 theme FUCAGP 1776:1781 arg1 levels 1783:1788 FUCAGP levels 1776:1788 FUCAGP levels measured during following-up of the patients after operation 1776:1849 Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention. 27295180 10 70 from patients 1438:1445 arg1 samples 1410:1416 serum samples 1404:1416 serum samples from various cancer patients 1404:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 10 70 from patients 1438:1445 arg1 levels 1384:1389 levels 1384:1389 levels of FUCAGP in serum samples from various cancer patients 1384:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 5 71 theme therapeutic 721:731 arg1 settings 733:740 therapeutic settings 721:740 therapeutic settings 721:740 In particular, there has been no validated biomarker for predicting treatment outcomes in therapeutic settings. 27295180 6 72 theme α1-acid 791:797 arg1 AGP 813:815 AGP 813:815 AGP 813:815 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 6 72 theme α1-acid 791:797 arg1 glycoprotein 799:810 serum α1-acid glycoprotein 785:810 serum α1-acid glycoprotein (AGP) 785:816 Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. 27295180 3 73 theme treatments 361:370 arg1 course 351:356 the time course 342:356 the time course of treatments 342:370 During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. 27295180 9 74 theme FUCAGP 1271:1276 arg1 levels 1261:1266 strikingly elevated levels 1241:1266 strikingly elevated levels of FUCAGP 1241:1276 Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. 27295180 2 75 theme neoadjuvant 282:292 arg1 chemotherapy 294:305 neoadjuvant chemotherapy 282:305 neoadjuvant chemotherapy 282:305 Additionally, neoadjuvant chemotherapy may be applied if required. 27295180 1 76 theme advanced-stage 146:159 arg1 cancer 161:166 advanced-stage cancer 146:166 advanced-stage cancer 146:166 One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. 27295180 12 77 theme relevant 1877:1884 arg1 biomarker 1886:1894 clinically relevant biomarker 1866:1894 clinically relevant biomarker of treatment intervention 1866:1920 Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention. 27295180 4 78 theme recurrence 516:525 arg1 evidence 504:511 early evidence 498:511 early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker 498:590 However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. 27295180 7 79 theme novel 993:997 arg1 software 999:1006 a novel software 991:1006 a novel software 991:1006 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 10 80 from levels 1384:1389 arg1 patients 1438:1445 various cancer patients 1423:1445 various cancer patients 1423:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 10 80 from levels 1384:1389 arg1 samples 1410:1416 serum samples 1404:1416 serum samples from various cancer patients 1404:1445 In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. 27295180 7 81 theme primary 912:918 arg1 structures 927:936 The primary glycan structures 908:936 The primary glycan structures of AGP 908:943 The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. 27295180 8 82 from abundance 1086:1094 arg1 AGP 1126:1128 AGP 1126:1128 AGP (FUCAGP) 1126:1137 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 27295180 8 82 from abundance 1086:1094 arg1 FUCAGP 1131:1136 FUCAGP 1131:1136 FUCAGP 1131:1136 Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. 25986781 2 0 theme protecting 535:544 arg1 group 546:550 a temporary protecting group 523:550 a temporary protecting group 523:550 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 0 theme protecting 535:544 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 0 theme protecting 535:544 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 1 dep high-yielding 1144:1156 arg1 % 1162:1162 ~90% 1159:1162 ~90% 1159:1162 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 2 2 theme -α-D-mannopyranosyl- 699:718 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 2 theme -α-D-mannopyranosyl- 699:718 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 3 theme 1→2 866:868 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 3 theme 1→2 866:868 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 0 4 dep structures 162:171 arg1 strategy 12:19 A synthetic strategy 0:19 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans 0:136 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 3 5 theme building 1100:1107 arg1 donors 1134:1139 donors 1134:1139 donors 1134:1139 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 3 5 theme building 1100:1107 arg1 blocks 1109:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 1 6 theme tri- 294:297 arg1 preparation 279:289 the preparation 275:289 the preparation 275:289 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 1 6 theme tri- 294:297 arg1 part 177:180 part 177:180 part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans 177:273 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 2 7 theme target 616:621 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 7 theme target 616:621 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 7 theme target 616:621 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 1 8 theme defined 393:399 arg1 structures 406:415 structurally defined part structures 380:415 structurally defined part structures of C. neoformans GXM capsular polysaccharide 380:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 3 9 theme part 1277:1280 arg1 structures 1282:1291 GXM polysaccharide part structures 1258:1291 GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates 1258:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 3 10 theme vaccine 1331:1337 arg1 candidates 1339:1348 vaccine candidates 1331:1348 vaccine candidates 1331:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 1 11 theme ongoing 188:194 arg1 project 196:202 an ongoing project 185:202 an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans 185:273 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 3 12 theme polysaccharide 1262:1275 arg1 structures 1282:1291 GXM polysaccharide part structures 1258:1291 GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates 1258:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 0 13 theme Cryptococcus 114:125 arg1 neoformans 127:136 Cryptococcus neoformans 114:136 Cryptococcus neoformans 114:136 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 1 14 theme part 401:404 arg1 structures 406:415 structurally defined part structures 380:415 structurally defined part structures of C. neoformans GXM capsular polysaccharide 380:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 2 15 theme 2-naphthalenylmethyl 678:697 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 15 theme 2-naphthalenylmethyl 678:697 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 16 used used 1126:1129 arg2 blocks 1109:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 3 16 used used 1126:1129 arg2 donors 1134:1139 donors 1134:1139 donors 1134:1139 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 4 17 theme suitable 1453:1460 arg1 acceptors 1443:1451 tri- and tetrasaccharide acceptors 1418:1451 tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks 1418:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 3 18 theme synthesized 1073:1083 arg1 donors 1134:1139 donors 1134:1139 donors 1134:1139 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 3 18 theme synthesized 1073:1083 arg1 blocks 1109:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 1 19 theme tetrasaccharide 303:317 arg1 preparation 279:289 the preparation 275:289 the preparation 275:289 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 1 19 theme tetrasaccharide 303:317 arg1 part 177:180 part 177:180 part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans 177:273 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 1 20 theme structures 406:415 arg1 construction 364:375 construction 364:375 construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide 364:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 3 21 theme GXM 1258:1260 arg1 structures 1282:1291 GXM polysaccharide part structures 1258:1291 GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates 1258:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 2 22 theme 1→2 764:766 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 22 theme 1→2 764:766 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 1 23 theme thioglycoside 319:331 arg1 blocks 342:347 thioglycoside building blocks 319:347 thioglycoside building blocks 319:347 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 0 24 theme capsular 138:145 arg1 structures 162:171 capsular polysaccharide structures 138:171 capsular polysaccharide structures 138:171 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 2 25 theme -[2,3,4-tri-O-benzyl-β-D-xylopyranosyl- 724:762 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 25 theme -[2,3,4-tri-O-benzyl-β-D-xylopyranosyl- 724:762 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 26 theme spacer-containing 1200:1216 arg1 acceptor 1218:1225 a spacer-containing acceptor 1198:1225 a spacer-containing acceptor to 1198:1228 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 2 27 theme -α-D-mannopyranosyl- 913:932 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 27 theme -α-D-mannopyranosyl- 913:932 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 1 28 theme project 196:202 arg1 preparation 279:289 the preparation 275:289 the preparation 275:289 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 1 28 theme project 196:202 arg1 part 177:180 part 177:180 part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans 177:273 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 2 29 theme trichloroacetimidate 556:575 arg1 donors 577:582 trichloroacetimidate donors 556:582 trichloroacetimidate donors 556:582 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 29 theme trichloroacetimidate 556:575 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 29 theme trichloroacetimidate 556:575 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 1 30 theme C. 420:421 arg1 polysaccharide 447:460 C. neoformans GXM capsular polysaccharide 420:460 C. neoformans GXM capsular polysaccharide 420:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 3 31 theme protein 1303:1309 arg1 conjugation 1311:1321 protein conjugation 1303:1321 protein conjugation to give vaccine candidates 1303:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 0 32 theme synthetic 2:10 arg1 strategy 12:19 A synthetic strategy 0:19 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans 0:136 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 2 33 theme temporary 525:533 arg1 group 546:550 a temporary protecting group 523:550 a temporary protecting group 523:550 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 33 theme temporary 525:533 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 33 theme temporary 525:533 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 34 theme 1→3 720:722 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 34 theme 1→3 720:722 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 35 theme 2-naphthalenylmethyl 892:911 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 35 theme 2-naphthalenylmethyl 892:911 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 4 36 theme thiosaccharide 1500:1513 arg1 blocks 1524:1529 larger thiosaccharide building blocks 1493:1529 larger thiosaccharide building blocks 1493:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 1 37 theme building 333:340 arg1 blocks 342:347 thioglycoside building blocks 319:347 thioglycoside building blocks 319:347 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 3 38 theme ready 1293:1297 arg1 structures 1282:1291 GXM polysaccharide part structures 1258:1291 GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates 1258:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 4 39 theme tri- 1418:1421 arg1 acceptors 1443:1451 tri- and tetrasaccharide acceptors 1418:1451 tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks 1418:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 4 40 theme further 1466:1472 arg1 elongation 1474:1483 further elongation 1466:1483 further elongation towards larger thiosaccharide building blocks 1466:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 4 41 theme larger 1493:1498 arg1 blocks 1524:1529 larger thiosaccharide building blocks 1493:1529 larger thiosaccharide building blocks 1493:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 2 42 theme naphthalenylmethyl 489:506 arg1 group 546:550 a temporary protecting group 523:550 a temporary protecting group 523:550 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 42 theme naphthalenylmethyl 489:506 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 42 theme naphthalenylmethyl 489:506 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 42 theme naphthalenylmethyl 489:506 arg1 donors 577:582 trichloroacetimidate donors 556:582 trichloroacetimidate donors 556:582 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 43 theme building 623:630 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 43 theme building 623:630 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 43 theme building 623:630 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 44 theme high-yielding 1144:1156 arg1 DMTST 1165:1169 high-yielding (~90%) DMTST 1144:1169 high-yielding (~90%) DMTST 1144:1169 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 1 45 theme vaccine 224:230 arg1 candidates 232:241 vaccine candidates 224:241 vaccine candidates 224:241 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 0 46 theme polysaccharide 147:160 arg1 structures 162:171 capsular polysaccharide structures 138:171 capsular polysaccharide structures 138:171 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 1 47 theme GXM 434:436 arg1 polysaccharide 447:460 C. neoformans GXM capsular polysaccharide 420:460 C. neoformans GXM capsular polysaccharide 420:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 0 48 theme thioglycoside 42:54 arg1 tri- 56:59 xylose-containing thioglycoside tri- 24:59 xylose-containing thioglycoside tri- 24:59 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 1 49 theme capsular 438:445 arg1 polysaccharide 447:460 C. neoformans GXM capsular polysaccharide 420:460 C. neoformans GXM capsular polysaccharide 420:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 2 50 theme 1→3 934:936 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 50 theme 1→3 934:936 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 0 51 theme xylose-containing 24:40 arg1 tri- 56:59 xylose-containing thioglycoside tri- 24:59 xylose-containing thioglycoside tri- 24:59 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 1 52 dep C. 420:421 arg1 neoformans 423:432 neoformans 423:432 neoformans 423:432 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 1 53 theme polysaccharide 447:460 arg1 structures 406:415 structurally defined part structures 380:415 structurally defined part structures of C. neoformans GXM capsular polysaccharide 380:460 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 4 54 theme building 1379:1386 arg1 blocks 1388:1393 the building blocks 1375:1393 the building blocks 1375:1393 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 1 55 theme Cryptococcus 251:262 arg1 neoformans 264:273 Cryptococcus neoformans 251:273 Cryptococcus neoformans 251:273 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 2 56 theme optimized 587:595 arg1 glycosylations 597:610 optimized glycosylations 587:610 optimized glycosylations 587:610 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 57 theme 6-O-acetyl-2,4-di-O-benzyl-3-O- 646:676 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 57 theme 6-O-acetyl-2,4-di-O-benzyl-3-O- 646:676 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 58 dep blocks 632:637 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 58 dep blocks 632:637 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 58 dep blocks 632:637 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 59 theme 2,3,4-tri-O-benzyl-β-D-xylopyranosyl- 828:864 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 59 theme 2,3,4-tri-O-benzyl-β-D-xylopyranosyl- 828:864 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 60 theme ethyl 640:644 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 60 theme ethyl 640:644 arg1 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -4,6-di-O-benzyl-1-thio-α-D-mannopyranoside 769:811 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 4 61 theme tetrasaccharide 1427:1441 arg1 acceptors 1443:1451 tri- and tetrasaccharide acceptors 1418:1451 tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks 1418:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 2 62 theme ethyl 822:826 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 62 theme ethyl 822:826 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 63 theme ~90 1159:1161 arg1 % 1162:1162 ~90% 1159:1162 ~90% 1159:1162 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 2 64 theme blocks 632:637 arg1 group 546:550 a temporary protecting group 523:550 a temporary protecting group 523:550 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 64 theme blocks 632:637 arg1 donors 577:582 trichloroacetimidate donors 556:582 trichloroacetimidate donors 556:582 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 64 theme blocks 632:637 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 64 theme blocks 632:637 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 1 65 dep tri- 294:297 arg1 blocks 342:347 thioglycoside building blocks 319:347 thioglycoside building blocks 319:347 As part of an ongoing project aimed at developing vaccine candidates against Cryptococcus neoformans the preparation of tri- and tetrasaccharide thioglycoside building blocks, to be used in construction of structurally defined part structures of C. neoformans GXM capsular polysaccharide, was investigated. 25986781 4 66 from groups 1365:1370 arg1 blocks 1388:1393 the building blocks 1375:1393 the building blocks 1375:1393 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 4 67 theme NAP 1361:1363 arg1 groups 1365:1370 the NAP groups 1357:1370 the NAP groups in the building blocks 1357:1393 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 3 68 dep promoted 1171:1178 arg1 afford 1251:1256 afford 1251:1256 afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates 1251:1348 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 0 69 theme building 81:88 arg1 blocks 90:95 tetrasaccharide building blocks 65:95 tetrasaccharide building blocks 65:95 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 2 70 theme -4,6-di-O-benzyl-3-O- 870:890 arg1 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2) 640:982 -[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl- 938:977 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 70 theme -4,6-di-O-benzyl-3-O- 870:890 arg1 blocks 632:637 the target building blocks 612:637 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 0 71 theme tetrasaccharide 65:79 arg1 blocks 90:95 tetrasaccharide building blocks 65:95 tetrasaccharide building blocks 65:95 A synthetic strategy to xylose-containing thioglycoside tri- and tetrasaccharide building blocks corresponding to Cryptococcus neoformans capsular polysaccharide structures. 25986781 4 72 theme building 1515:1522 arg1 blocks 1524:1529 larger thiosaccharide building blocks 1493:1529 larger thiosaccharide building blocks 1493:1529 Also, the NAP groups in the building blocks were removed to obtain tri- and tetrasaccharide acceptors suitable for further elongation towards larger thiosaccharide building blocks. 25986781 2 73 theme NAP 509:511 arg1 group 546:550 a temporary protecting group 523:550 a temporary protecting group 523:550 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 73 theme NAP 509:511 arg1 ether 514:518 a naphthalenylmethyl (NAP) ether 487:518 a naphthalenylmethyl (NAP) ether 487:518 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 73 theme NAP 509:511 arg1 -6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside 984:1032 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21) 612:1037 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 2 73 theme NAP 509:511 arg1 donors 577:582 trichloroacetimidate donors 556:582 trichloroacetimidate donors 556:582 Using a naphthalenylmethyl (NAP) ether as a temporary protecting group and trichloroacetimidate donors in optimized glycosylations the target building blocks, ethyl 6-O-acetyl-2,4-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)]-4,6-di-O-benzyl-1-thio-α-D-mannopyranoside (16) and ethyl 2,3,4-tri-O-benzyl-β-D-xylopyranosyl-(1→2)-4,6-di-O-benzyl-3-O-(2-naphthalenylmethyl)-α-D-mannopyranosyl-(1→3)-[2,3,4-tri-O-benzyl-β-D-xylopyra-nosyl-(1→2)]-6-O-acetyl-4-O-benzyl-1-thio-α-D-mannopyranoside (21), were efficiently prepared. 25986781 3 74 theme thiosaccharide 1085:1098 arg1 donors 1134:1139 donors 1134:1139 donors 1134:1139 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 25986781 3 74 theme thiosaccharide 1085:1098 arg1 blocks 1109:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks 1067:1114 These synthesized thiosaccharide building blocks were then used as donors in high-yielding (~90%) DMTST promoted glycosylations to a spacer-containing acceptor to, after deprotection, afford GXM polysaccharide part structures ready for protein conjugation to give vaccine candidates. 26055112 10 0 dep complete 1829:1836 arg1 provide 1898:1904 provide 1898:1904 provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F 1898:2042 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 9 1 theme types 1747:1751 arg1 members 1656:1662 the members 1652:1662 the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types 1652:1751 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 10 2 theme determined 1768:1777 arg1 structures 1779:1788 The presently determined structures 1754:1788 The presently determined structures of S. pneumoniae serotypes 35F and 35C 1754:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 6 3 from CPS35F 1123:1128 arg1 distinct 1102:1109 distinct 1102:1109 distinct 1102:1109 Moreover, CPS35C is distinct from either CPS35F or CPS35B but closely related to CPS35A and identical to de-O-acetylated CPS42. 26055112 7 4 theme cross-reactive 1353:1366 arg1 serotypes 1368:1376 other cross-reactive serotypes 1347:1376 other cross-reactive serotypes 1347:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 11 5 theme immune 2201:2206 arg1 selection 2208:2216 immune selection 2201:2216 immune selection 2201:2216 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 6 6 from CPS35B 1133:1138 arg1 distinct 1102:1109 distinct 1102:1109 distinct 1102:1109 Moreover, CPS35C is distinct from either CPS35F or CPS35B but closely related to CPS35A and identical to de-O-acetylated CPS42. 26055112 4 7 theme magnetic 749:756 arg1 resonance 758:766 heteronuclear magnetic resonance 735:766 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 4 7 theme magnetic 749:756 arg1 NMR 769:771 NMR 769:771 NMR 769:771 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 10 8 theme serogroup 1975:1983 arg1 members 1959:1965 different members 1949:1965 different members of this serogroup 1949:1983 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 9 theme pneumoniae 1796:1805 arg1 serotypes 1807:1815 S. pneumoniae serotypes 35F and 35C 1793:1827 S. pneumoniae serotypes 35F and 35C 1793:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 9 theme pneumoniae 1796:1805 arg1 35C 1825:1827 35C 1825:1827 35C 1825:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 9 theme pneumoniae 1796:1805 arg1 35F 1817:1819 35F 1817:1819 35F 1817:1819 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 7 10 theme serogroup 1330:1338 arg1 serotypes 1368:1376 other cross-reactive serotypes 1347:1376 other cross-reactive serotypes 1347:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 7 10 theme serogroup 1330:1338 arg1 members 1315:1321 the members 1311:1321 the members of CPS serogroup 35 1311:1341 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 0 11 theme Other 151:155 arg1 Serotypes 172:180 Other Cross-Reactive Serotypes 151:180 Other Cross-Reactive Serotypes 151:180 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 7 12 theme genetic 1274:1280 arg1 relations 1282:1290 the structural and genetic relations 1255:1290 the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes 1255:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 11 13 theme polysaccharide 2149:2162 arg1 subgroups 2164:2172 three distinct capsular polysaccharide subgroups 2125:2172 three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host 2125:2234 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 4 14 theme repeat 845:850 arg1 structures 857:866 the following repeat unit structures 831:866 the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated 831:920 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 7 15 theme structural 1259:1268 arg1 relations 1282:1290 the structural and genetic relations 1255:1290 the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes 1255:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 11 16 theme distinct 2131:2138 arg1 subgroups 2164:2172 three distinct capsular polysaccharide subgroups 2125:2172 three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host 2125:2234 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 10 17 theme serogroup 1873:1881 arg1 characterization 1853:1868 the structural characterization 1838:1868 the structural characterization of serogroup 35 1838:1884 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 1 18 theme polysaccharides 246:260 arg1 structures 198:207 The structures 194:207 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) 183:267 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 1 18 theme polysaccharides 246:260 arg1 essential 273:281 essential 273:281 essential 273:281 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 9 19 theme serotypes 1628:1636 arg1 group 1608:1612 a group 1606:1612 a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types 1606:1751 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 19 theme serotypes 1628:1636 arg1 serotypes 1628:1636 nonvaccine serotypes 1617:1636 nonvaccine serotypes 1617:1636 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 35A 1698:1700 35A 1698:1700 35A 1698:1700 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 i.e. 1681:1684 i.e. 1681:1684 i.e. 1681:1684 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 35B 1703:1705 35B 1703:1705 35B 1703:1705 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 types 1687:1691 types 35F, 35A, 35B, and 35C 1687:1714 types 35F, 35A, 35B, and 35C 1687:1714 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 35C 1712:1714 35C 1712:1714 35C 1712:1714 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 9 20 dep types 1687:1691 arg1 35F 1693:1695 35F 1693:1695 35F 1693:1695 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 7 21 theme comprehensive 1233:1245 arg1 view 1247:1250 a comprehensive view 1231:1250 a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes 1231:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 5 22 theme immunizing 948:957 arg1 serotype 959:966 the immunizing serotype 944:966 the immunizing serotype for the production of group 35 serum 944:1003 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 5 22 theme immunizing 948:957 arg1 CPS35F 936:941 CPS35F 936:941 CPS35F 936:941 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 3 23 theme serotype 560:567 arg1 CPS35A 574:579 CPS35A 574:579 CPS35A 574:579 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 3 23 theme serotype 560:567 arg1 35A 569:571 CPS serotype 35A 556:571 CPS serotype 35A (CPS35A) 556:580 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 9 24 theme serogroup 1667:1675 arg1 members 1656:1662 the members 1652:1662 the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types 1652:1751 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 8 25 theme IMPORTANCE 1379:1388 arg1 Cross-reactions 1390:1404 IMPORTANCE Cross-reactions 1379:1404 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes 1379:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 0 26 theme pneumoniae 77:86 arg1 Structures 0:9 Structures 0:9 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae 0:86 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 10 27 theme first 1910:1914 arg1 description 1930:1940 the first comprehensive description 1906:1940 the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F 1906:2042 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 5 28 theme group 990:994 arg1 serum 999:1003 group 35 serum 990:1003 group 35 serum 990:1003 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 2 29 theme related 495:501 arg1 42 520:521 42 520:521 42 520:521 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 29 theme related 495:501 arg1 47F 527:529 47F 527:529 47F 527:529 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 29 theme related 495:501 arg1 34 516:517 34 516:517 34 516:517 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 29 theme related 495:501 arg1 type 503:506 genetically related type 20 483:509 genetically related type 20 483:509 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 29 theme related 495:501 arg1 29 512:513 29 512:513 29 512:513 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 8 30 theme rabbit 1420:1425 arg1 antisera 1427:1434 diagnostic rabbit antisera 1409:1434 diagnostic rabbit antisera 1409:1434 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 11 31 theme human 2225:2229 arg1 host 2231:2234 the human host 2221:2234 the human host 2221:2234 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 0 32 theme Magnetic 110:117 arg1 Resonance 119:127 Nuclear Magnetic Resonance 102:127 Nuclear Magnetic Resonance 102:127 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 5 33 theme serum 999:1003 arg1 production 976:985 the production 972:985 the production of group 35 serum 972:1003 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 8 34 theme polysaccharide 1475:1488 arg1 serotypes 1490:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 8 35 with Cross-reactions 1390:1404 arg1 serotypes 1490:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 1 36 theme antigenic 300:308 arg1 relationships 329:341 the antigenic as well as genetic relationships 296:341 the antigenic as well as genetic relationships between CPS serotypes 296:363 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 4 37 theme present 647:653 arg1 study 655:659 the present study 643:659 the present study 643:659 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 0 38 theme Capsular 14:21 arg1 Polysaccharide 23:36 Capsular Polysaccharide Serotypes 35F and 35C 14:58 Capsular Polysaccharide Serotypes 35F and 35C 14:58 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 5 39 theme serogroup 1068:1076 arg1 members 1057:1063 other members 1051:1063 other members of serogroup 35 1051:1079 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 10 40 dep serotypes 1807:1815 arg1 serotypes 1807:1815 S. pneumoniae serotypes 35F and 35C 1793:1827 S. pneumoniae serotypes 35F and 35C 1793:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 40 dep serotypes 1807:1815 arg1 35C 1825:1827 35C 1825:1827 35C 1825:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 40 dep serotypes 1807:1815 arg1 35F 1817:1819 35F 1817:1819 35F 1817:1819 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 2 41 dep types 423:427 arg1 35A 434:436 35A 434:436 35A 434:436 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 41 dep types 423:427 arg1 i.e. 417:420 i.e. 417:420 i.e. 417:420 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 41 dep types 423:427 arg1 35B 439:441 35B 439:441 35B 439:441 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 41 dep types 423:427 arg1 types 423:427 types 35F, 35A, 35B, and 35C 423:450 types 35F, 35A, 35B, and 35C 423:450 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 41 dep types 423:427 arg1 35F 429:431 35F 429:431 35F 429:431 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 41 dep types 423:427 arg1 35C 448:450 35C 448:450 35C 448:450 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 4 42 dep spectroscopy 774:785 arg1 analyses 812:819 analyses 812:819 analyses 812:819 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 1 43 theme genetic 321:327 arg1 relationships 329:341 the antigenic as well as genetic relationships 296:341 the antigenic as well as genetic relationships between CPS serotypes 296:363 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 4 44 theme high-resolution 719:733 arg1 spectroscopy 774:785 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 0 45 dep Serotypes 38:46 arg1 35F 48:50 35F 48:50 35F 48:50 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 0 45 dep Serotypes 38:46 arg1 Serotypes 38:46 Serotypes 35F and 35C 38:58 Capsular Polysaccharide Serotypes 35F and 35C 14:58 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 0 45 dep Serotypes 38:46 arg1 35C 56:58 35C 56:58 35C 56:58 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 4 46 theme CPS35F 679:684 arg1 serotypes 666:674 the serotypes 662:674 the serotypes of CPS35F and CPS35C 662:695 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 8 47 dep Streptococcus 1441:1453 arg1 pneumoniae 1455:1464 pneumoniae 1455:1464 pneumoniae 1455:1464 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 9 48 theme other 1721:1725 arg1 types 1747:1751 other genetically related types 1721:1751 other genetically related types 1721:1751 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 3 49 theme CPS35B 586:591 arg1 structures 542:551 the structures 538:551 the structures of CPS serotype 35A (CPS35A) and CPS35B 538:591 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 4 50 theme CPS35C 690:695 arg1 serotypes 666:674 the serotypes 662:674 the serotypes of CPS35F and CPS35C 662:695 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 2 51 theme CPS 399:401 arg1 types 423:427 types 35F, 35A, 35B, and 35C 423:450 types 35F, 35A, 35B, and 35C 423:450 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 2 51 theme CPS 399:401 arg1 serogroup 403:411 CPS serogroup 35 399:414 CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) 399:451 The four serotypes that comprise CPS serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) are known to cross-react with genetically related type 20, 29, 34, 42, or 47F. 26055112 9 52 theme related 1739:1745 arg1 types 1747:1751 other genetically related types 1721:1751 other genetically related types 1721:1751 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 7 53 theme other 1347:1351 arg1 serotypes 1368:1376 other cross-reactive serotypes 1347:1376 other cross-reactive serotypes 1347:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 8 54 theme serogroup 1545:1553 arg1 members 1525:1531 members 1525:1531 members of the same serogroup 1525:1553 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 10 55 theme serotypes 1807:1815 arg1 structures 1779:1788 The presently determined structures 1754:1788 The presently determined structures of S. pneumoniae serotypes 35F and 35C 1754:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 1 56 theme UNLABELLED 183:192 arg1 structures 198:207 The structures 194:207 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) 183:267 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 1 56 theme UNLABELLED 183:192 arg1 essential 273:281 essential 273:281 essential 273:281 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 7 57 theme CPS 1326:1328 arg1 serogroup 1330:1338 CPS serogroup 35 1326:1341 CPS serogroup 35 1326:1341 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 4 58 theme heteronuclear 735:747 arg1 resonance 758:766 heteronuclear magnetic resonance 735:766 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 4 58 theme heteronuclear 735:747 arg1 NMR 769:771 NMR 769:771 NMR 769:771 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 10 59 theme S. 1793:1794 arg1 serotypes 1807:1815 S. pneumoniae serotypes 35F and 35C 1793:1827 S. pneumoniae serotypes 35F and 35C 1793:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 59 theme S. 1793:1794 arg1 35C 1825:1827 35C 1825:1827 35C 1825:1827 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 59 theme S. 1793:1794 arg1 35F 1817:1819 35F 1817:1819 35F 1817:1819 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 4 60 theme resonance 758:766 arg1 spectroscopy 774:785 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 high-resolution heteronuclear magnetic resonance (NMR) spectroscopy 719:785 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 11 61 theme capsular 2140:2147 arg1 polysaccharide 2149:2162 capsular polysaccharide 2140:2162 three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host 2125:2234 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 4 62 dep [Formula 869:876 arg1 see 879:881 see 879:881 see text 879:886 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 1 63 theme Streptococcus 212:224 arg1 CPSs 263:266 CPSs 263:266 CPSs 263:266 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 1 63 theme Streptococcus 212:224 arg1 polysaccharides 246:260 Streptococcus pneumoniae capsular polysaccharides 212:260 Streptococcus pneumoniae capsular polysaccharides (CPSs) 212:267 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 7 64 theme relations 1282:1290 arg1 view 1247:1250 a comprehensive view 1231:1250 a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes 1231:1376 The findings provide a comprehensive view of the structural and genetic relations that exist between the members of CPS serogroup 35 and other cross-reactive serotypes. 26055112 4 65 dep structures 857:866 arg1 [Formula 869:876 [Formula 869:876 the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated 831:920 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 1 66 theme capsular 237:244 arg1 CPSs 263:266 CPSs 263:266 CPSs 263:266 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 1 66 theme capsular 237:244 arg1 polysaccharides 246:260 Streptococcus pneumoniae capsular polysaccharides 212:260 Streptococcus pneumoniae capsular polysaccharides (CPSs) 212:267 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 9 67 theme nonvaccine 1617:1626 arg1 serotypes 1628:1636 nonvaccine serotypes 1617:1636 nonvaccine serotypes 1617:1636 Exceptions do, however, occur, most notably among a group of nonvaccine serotypes that includes the members of serogroup 35 (i.e., types 35F, 35A, 35B, and 35C) and other genetically related types. 26055112 1 68 theme CPS 351:353 arg1 serotypes 355:363 CPS serotypes 351:363 CPS serotypes 351:363 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 6 69 theme de-O-acetylated 1187:1201 arg1 CPS42 1203:1207 de-O-acetylated CPS42 1187:1207 de-O-acetylated CPS42 1187:1207 Moreover, CPS35C is distinct from either CPS35F or CPS35B but closely related to CPS35A and identical to de-O-acetylated CPS42. 26055112 4 70 theme glycosyl 791:798 arg1 composition 800:810 glycosyl composition 791:810 glycosyl composition 791:810 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 10 71 theme structural 1842:1851 arg1 characterization 1853:1868 the structural characterization 1838:1868 the structural characterization of serogroup 35 1838:1884 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 3 72 theme CPS 556:558 arg1 CPS35A 574:579 CPS35A 574:579 CPS35A 574:579 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 3 72 theme CPS 556:558 arg1 35A 569:571 CPS serotype 35A 556:571 CPS serotype 35A (CPS35A) 556:580 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 11 73 theme genetic 2064:2070 arg1 features 2072:2079 The structural and genetic features 2045:2079 The structural and genetic features of these serotypes 2045:2098 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 11 74 theme serotypes 2090:2098 arg1 features 2072:2079 The structural and genetic features 2045:2079 The structural and genetic features of these serotypes 2045:2098 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 11 75 theme structural 2049:2058 arg1 features 2072:2079 The structural and genetic features 2045:2079 The structural and genetic features of these serotypes 2045:2098 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 8 76 theme diagnostic 1409:1418 arg1 antisera 1427:1434 diagnostic rabbit antisera 1409:1434 diagnostic rabbit antisera 1409:1434 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 10 77 theme comprehensive 1916:1928 arg1 description 1930:1940 the first comprehensive description 1906:1940 the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F 1906:2042 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 0 78 theme Nuclear 102:108 arg1 Resonance 119:127 Nuclear Magnetic Resonance 102:127 Nuclear Magnetic Resonance 102:127 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 4 79 theme unit 852:855 arg1 structures 857:866 the following repeat unit structures 831:866 the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated 831:920 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 8 80 theme antisera 1427:1434 arg1 Cross-reactions 1390:1404 IMPORTANCE Cross-reactions 1379:1404 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes 1379:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 11 81 theme subgroups 2164:2172 arg1 existence 2112:2120 the existence 2108:2120 the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host 2108:2234 The structural and genetic features of these serotypes suggest the existence of three distinct capsular polysaccharide subgroups that presumably emerged by immune selection in the human host. 26055112 8 82 theme Streptococcus 1441:1453 arg1 serotypes 1490:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 8 83 theme capsular 1466:1473 arg1 serotypes 1490:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 Streptococcus pneumoniae capsular polysaccharide serotypes 1441:1498 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 26055112 0 84 theme Polysaccharide 23:36 arg1 Structures 0:9 Structures 0:9 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae 0:86 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 1 85 dep Streptococcus 212:224 arg1 pneumoniae 226:235 pneumoniae 226:235 pneumoniae 226:235 UNLABELLED The structures of Streptococcus pneumoniae capsular polysaccharides (CPSs) are essential for defining the antigenic as well as genetic relationships between CPS serotypes. 26055112 0 86 theme Cross-Reactive 157:170 arg1 Serotypes 172:180 Other Cross-Reactive Serotypes 151:180 Other Cross-Reactive Serotypes 151:180 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 3 87 theme 35A 569:571 arg1 structures 542:551 the structures 538:551 the structures of CPS serotype 35A (CPS35A) and CPS35B 538:591 While the structures of CPS serotype 35A (CPS35A) and CPS35B are known, those of CPS35F and CPS35C are not. 26055112 4 88 theme following 835:843 arg1 structures 857:866 the following repeat unit structures 831:866 the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated 831:920 In the present study, the serotypes of CPS35F and CPS35C were characterized by high-resolution heteronuclear magnetic resonance (NMR) spectroscopy and glycosyl composition analyses to reveal the following repeat unit structures: [Formula: see text] where OAc indicates O-acetylated. 26055112 5 89 theme other 1051:1055 arg1 members 1057:1063 other members 1051:1063 other members of serogroup 35 1051:1079 Importantly, CPS35F, the immunizing serotype for the production of group 35 serum, more closely resembles CPS34 and CPS47F than other members of serogroup 35. 26055112 0 90 dep Polysaccharide 23:36 arg1 35F 48:50 35F 48:50 35F 48:50 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 0 90 dep Polysaccharide 23:36 arg1 Serotypes 38:46 Serotypes 35F and 35C 38:58 Capsular Polysaccharide Serotypes 35F and 35C 14:58 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 0 90 dep Polysaccharide 23:36 arg1 35C 56:58 35C 56:58 35C 56:58 Structures of Capsular Polysaccharide Serotypes 35F and 35C of Streptococcus pneumoniae Determined by Nuclear Magnetic Resonance and Their Relation to Other Cross-Reactive Serotypes. 26055112 10 91 dep types 2018:2022 arg1 42 2032:2033 42 2032:2033 42 2032:2033 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 91 dep types 2018:2022 arg1 47F 2040:2042 47F 2040:2042 47F 2040:2042 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 91 dep types 2018:2022 arg1 34 2028:2029 34 2028:2029 34 2028:2029 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 91 dep types 2018:2022 arg1 29 2024:2025 29 2024:2025 29 2024:2025 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 10 92 theme different 1949:1957 arg1 members 1959:1965 different members 1949:1965 different members of this serogroup 1949:1983 The presently determined structures of S. pneumoniae serotypes 35F and 35C complete the structural characterization of serogroup 35 and thereby provide the first comprehensive description of how different members of this serogroup are related to each other and to types 29, 34, 42, and 47F. 26055112 8 93 theme same 1540:1543 arg1 serogroup 1545:1553 the same serogroup 1536:1553 the same serogroup 1536:1553 IMPORTANCE Cross-reactions of diagnostic rabbit antisera with Streptococcus pneumoniae capsular polysaccharide serotypes are generally limited to members of the same serogroup. 28905280 0 0 theme disaccharide 116:127 arg1 structures 129:138 novel disaccharide structures 110:138 novel disaccharide structures 110:138 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 4 1 theme produced 568:575 arg1 disaccharides 594:606 the produced galactose-fucose disaccharides 564:606 the produced galactose-fucose disaccharides 564:606 The reaction product was analyzed and the produced galactose-fucose disaccharides were purified. 28905280 7 2 theme oligosaccharides 1064:1079 arg1 disaccharides 1032:1044 40% disaccharides 1028:1044 40% disaccharides 1028:1044 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 2 theme oligosaccharides 1064:1079 arg1 % 1052:1052 18% 1050:1052 18% of larger oligosaccharides 1050:1079 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 2 theme oligosaccharides 1064:1079 arg1 oligosaccharides 1064:1079 larger oligosaccharides 1057:1079 larger oligosaccharides 1057:1079 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 2 theme oligosaccharides 1064:1079 arg1 monosaccharides 1011:1025 42% total monosaccharides 1001:1025 42% total monosaccharides 1001:1025 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 9 3 from oligosaccharides 1220:1235 arg1 % 1257:1257 44% 1255:1257 44% 1255:1257 The fraction of fGOS from total oligosaccharides was determined as 44%. 28905280 9 3 from oligosaccharides 1220:1235 arg1 fraction 1192:1199 The fraction 1188:1199 The fraction of fGOS from total oligosaccharides 1188:1235 The fraction of fGOS from total oligosaccharides was determined as 44%. 28905280 0 4 theme novel 110:114 arg1 structures 129:138 novel disaccharide structures 110:138 novel disaccharide structures 110:138 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 9 5 theme total 1214:1218 arg1 oligosaccharides 1220:1235 total oligosaccharides 1214:1235 total oligosaccharides 1214:1235 The fraction of fGOS from total oligosaccharides was determined as 44%. 28905280 5 6 theme structure 739:747 arg1 Galβ1-3Fuc 749:758 the structure Galβ1-3Fuc 735:758 the structure Galβ1-3Fuc 735:758 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 2 7 from fucose 401:406 arg1 β-galactosidase 297:311 a commercial β-galactosidase 284:311 a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 284:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 2 7 from fucose 401:406 arg1 production 320:329 the production 316:329 the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 316:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 1 8 theme healthy 210:216 arg1 system 225:230 a healthy immune system 208:230 a healthy immune system 208:230 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 2 9 from lactose 389:395 arg1 β-galactosidase 297:311 a commercial β-galactosidase 284:311 a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 284:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 2 9 from lactose 389:395 arg1 production 320:329 the production 316:329 the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 316:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 1 10 contain have 170:173 arg2 role 188:191 an important role 175:191 an important role 175:191 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 1 10 contain have 170:173 arg1 oligosaccharides 153:168 Fucosylated oligosaccharides 141:168 Fucosylated oligosaccharides 141:168 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 7 11 contain contained 991:999 arg1 product 983:989 the final product 973:989 the final product 973:989 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 11 contain contained 991:999 arg2 monosaccharides 1011:1025 42% total monosaccharides 1001:1025 42% total monosaccharides 1001:1025 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 11 contain contained 991:999 arg2 % 1052:1052 18% 1050:1052 18% of larger oligosaccharides 1050:1079 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 11 contain contained 991:999 arg2 disaccharides 1032:1044 40% disaccharides 1028:1044 40% disaccharides 1028:1044 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 11 contain contained 991:999 arg2 oligosaccharides 1064:1079 larger oligosaccharides 1057:1079 larger oligosaccharides 1057:1079 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 7 12 theme final 977:981 arg1 product 983:989 the final product 973:989 the final product 973:989 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 0 13 theme structures 129:138 arg1 identification 92:105 identification 92:105 identification 92:105 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 0 13 theme structures 129:138 arg1 β-galactosidase 72:86 β-galactosidase 72:86 β-galactosidase 72:86 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 8 14 theme 290 1082:1084 arg1 μmol 1086:1089 290 μmol 1082:1089 290 μmol of fGOS 1082:1097 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 2 15 theme galacto-oligosaccharides 352:375 arg1 production 320:329 the production 316:329 the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 316:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 4 16 theme reaction 530:537 arg1 product 539:545 The reaction product 526:545 The reaction product 526:545 The reaction product was analyzed and the produced galactose-fucose disaccharides were purified. 28905280 10 17 theme putative 1301:1308 arg1 route 1310:1314 a new putative route 1295:1314 a new putative route to deliver fucose to the intestine 1295:1349 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 10 17 theme putative 1301:1308 arg1 product 1270:1276 This fGOS product 1260:1276 This fGOS product 1260:1276 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 8 18 theme mixture 1133:1139 arg1 gram 1116:1119 gram 1116:1119 gram of reaction mixture 1116:1139 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 6 19 theme more 895:898 arg1 detail 900:905 more detail 895:905 more detail 895:905 Additionally, the product composition was defined in more detail using several different analytical methods. 28905280 2 20 theme fucose-containing 334:350 arg1 galacto-oligosaccharides 352:375 fucose-containing galacto-oligosaccharides 334:375 fucose-containing galacto-oligosaccharides (fGOS) 334:382 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 2 20 theme fucose-containing 334:350 arg1 fGOS 378:381 fGOS 378:381 fGOS 378:381 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 1 21 theme immune 218:223 arg1 system 225:230 a healthy immune system 208:230 a healthy immune system 208:230 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 5 22 theme major 716:720 arg1 product 722:728 one major product 712:728 one major product with the structure Galβ1-3Fuc 712:758 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 7 23 theme % 1030:1030 arg1 disaccharides 1032:1044 40% disaccharides 1028:1044 40% disaccharides 1028:1044 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 3 24 theme optimal 466:472 arg1 conditions 474:483 optimal conditions 466:483 optimal conditions 466:483 The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. 28905280 5 25 dep structures 792:801 arg1 structures 792:801 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 25 dep structures 792:801 arg1 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 25 dep structures 792:801 arg1 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 8 26 theme fucose 1162:1167 arg1 % 1147:1147 37% 1145:1147 37% of the added fucose 1145:1167 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 8 26 theme fucose 1162:1167 arg1 fucose 1162:1167 the added fucose 1152:1167 the added fucose 1152:1167 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 0 27 theme Enzymatic 0:8 arg1 synthesis 10:18 Enzymatic synthesis 0:18 Enzymatic synthesis of fucose-containing galacto-oligosaccharides 0:64 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 1 28 theme homeostatic 236:246 arg1 microflora 252:261 homeostatic gut microflora 236:261 homeostatic gut microflora 236:261 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 3 29 theme experiment 444:453 arg1 design 455:460 experiment design 444:460 experiment design 444:460 The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. 28905280 7 30 theme 40 1028:1029 arg1 % 1030:1030 % 1030:1030 % 1030:1030 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 1 31 theme Fucosylated 141:151 arg1 oligosaccharides 153:168 Fucosylated oligosaccharides 141:168 Fucosylated oligosaccharides 141:168 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 2 32 from β-galactosidase 297:311 arg1 production 320:329 the production 316:329 the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 316:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 2 32 from β-galactosidase 297:311 arg1 fucose 401:406 fucose 401:406 fucose 401:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 2 32 from β-galactosidase 297:311 arg1 lactose 389:395 lactose 389:395 lactose 389:395 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 1 33 theme gut 248:250 arg1 microflora 252:261 homeostatic gut microflora 236:261 homeostatic gut microflora 236:261 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 8 34 theme added 1156:1160 arg1 fucose 1162:1167 the added fucose 1152:1167 the added fucose 1152:1167 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 2 35 theme commercial 286:295 arg1 β-galactosidase 297:311 a commercial β-galactosidase 284:311 a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose 284:406 This study employed a commercial β-galactosidase in the production of fucose-containing galacto-oligosaccharides (fGOS) from lactose and fucose. 28905280 10 36 theme fGOS 1265:1268 arg1 product 1270:1276 This fGOS product 1260:1276 This fGOS product 1260:1276 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 10 36 theme fGOS 1265:1268 arg1 route 1310:1314 a new putative route 1295:1314 a new putative route to deliver fucose to the intestine 1295:1349 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 0 37 theme galacto-oligosaccharides 41:64 arg1 synthesis 10:18 Enzymatic synthesis 0:18 Enzymatic synthesis of fucose-containing galacto-oligosaccharides 0:64 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 3 38 theme batch 491:495 arg1 production 497:506 a batch production 489:506 a batch production in 3-liter scale 489:523 The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. 28905280 0 39 theme fucose-containing 23:39 arg1 galacto-oligosaccharides 41:64 fucose-containing galacto-oligosaccharides 23:64 fucose-containing galacto-oligosaccharides 23:64 Enzymatic synthesis of fucose-containing galacto-oligosaccharides using β-galactosidase and identification of novel disaccharide structures. 28905280 6 40 theme product 860:866 arg1 composition 868:878 the product composition 856:878 the product composition 856:878 Additionally, the product composition was defined in more detail using several different analytical methods. 28905280 3 41 theme 3-liter 511:517 arg1 scale 519:523 3-liter scale 511:523 3-liter scale 511:523 The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. 28905280 5 42 with product 722:728 arg1 structures 792:801 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 42 with product 722:728 arg1 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 42 with product 722:728 arg1 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 42 with product 722:728 arg1 Galβ1-3Fuc 749:758 the structure Galβ1-3Fuc 735:758 the structure Galβ1-3Fuc 735:758 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 43 theme minor 768:772 arg1 products 774:781 two minor products 764:781 two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc 764:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 44 with products 774:781 arg1 structures 792:801 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 the structures Galβ1-4Fuc and Galβ1-2Fuc 788:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 44 with products 774:781 arg1 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 Galβ1-2Fuc 818:827 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 44 with products 774:781 arg1 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 Galβ1-4Fuc 803:812 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 44 with products 774:781 arg1 Galβ1-3Fuc 749:758 the structure Galβ1-3Fuc 735:758 the structure Galβ1-3Fuc 735:758 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 5 45 theme disaccharides 647:659 arg1 structures 627:636 The structures 623:636 The structures of these disaccharides 623:659 The structures of these disaccharides were determined using NMR and it was verified that one major product with the structure Galβ1-3Fuc and two minor products with the structures Galβ1-4Fuc and Galβ1-2Fuc were formed. 28905280 8 46 theme reaction 1124:1131 arg1 mixture 1133:1139 reaction mixture 1124:1139 reaction mixture 1124:1139 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 7 47 theme total 1005:1009 arg1 monosaccharides 1011:1025 42% total monosaccharides 1001:1025 42% total monosaccharides 1001:1025 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 8 48 theme fGOS 1094:1097 arg1 μmol 1086:1089 290 μmol 1082:1089 290 μmol of fGOS 1082:1097 290 μmol of fGOS was produced per gram of reaction mixture and 37% of the added fucose was bound to fGOS. 28905280 6 49 theme several 913:919 arg1 methods 942:948 several different analytical methods 913:948 several different analytical methods 913:948 Additionally, the product composition was defined in more detail using several different analytical methods. 28905280 1 50 theme important 178:186 arg1 role 188:191 an important role 175:191 an important role 175:191 Fucosylated oligosaccharides have an important role in maintaining a healthy immune system and homeostatic gut microflora. 28905280 6 51 theme analytical 931:940 arg1 methods 942:948 several different analytical methods 913:948 several different analytical methods 913:948 Additionally, the product composition was defined in more detail using several different analytical methods. 28905280 10 52 used used 1287:1290 arg2 route 1310:1314 a new putative route 1295:1314 a new putative route to deliver fucose to the intestine 1295:1349 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 10 52 used used 1287:1290 arg2 product 1270:1276 This fGOS product 1260:1276 This fGOS product 1260:1276 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 3 53 from production 497:506 arg1 scale 519:523 3-liter scale 511:523 3-liter scale 511:523 The production was optimized using experiment design and optimal conditions for a batch production in 3-liter scale. 28905280 7 54 theme 42 1001:1002 arg1 % 1003:1003 % 1003:1003 % 1003:1003 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 6 55 theme different 921:929 arg1 methods 942:948 several different analytical methods 913:948 several different analytical methods 913:948 Additionally, the product composition was defined in more detail using several different analytical methods. 28905280 9 56 theme fGOS 1204:1207 arg1 % 1257:1257 44% 1255:1257 44% 1255:1257 The fraction of fGOS from total oligosaccharides was determined as 44%. 28905280 9 56 theme fGOS 1204:1207 arg1 fraction 1192:1199 The fraction 1188:1199 The fraction of fGOS from total oligosaccharides 1188:1235 The fraction of fGOS from total oligosaccharides was determined as 44%. 28905280 7 57 theme % 1003:1003 arg1 monosaccharides 1011:1025 42% total monosaccharides 1001:1025 42% total monosaccharides 1001:1025 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 4 58 theme galactose-fucose 577:592 arg1 disaccharides 594:606 the produced galactose-fucose disaccharides 564:606 the produced galactose-fucose disaccharides 564:606 The reaction product was analyzed and the produced galactose-fucose disaccharides were purified. 28905280 7 59 theme larger 1057:1062 arg1 oligosaccharides 1064:1079 larger oligosaccharides 1057:1079 larger oligosaccharides 1057:1079 It was concluded that the final product contained 42% total monosaccharides, 40% disaccharides and 18% of larger oligosaccharides. 28905280 10 60 theme new 1297:1299 arg1 route 1310:1314 a new putative route 1295:1314 a new putative route to deliver fucose to the intestine 1295:1349 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 28905280 10 60 theme new 1297:1299 arg1 product 1270:1276 This fGOS product 1260:1276 This fGOS product 1260:1276 This fGOS product could be used as a new putative route to deliver fucose to the intestine. 26169737 0 0 theme Mass 79:82 arg1 Spectrometry 84:95 Negative Ion Mass Spectrometry 66:95 Negative Ion Mass Spectrometry 66:95 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 0 1 theme Ion 101:103 arg1 Mobility 105:112 Ion Mobility 101:112 Ion Mobility 101:112 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 0 2 from Determination 0:12 arg1 Glycoproteins 49:61 Viral Glycoproteins 43:61 Viral Glycoproteins 43:61 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 0 3 link N-linked 17:24 arg1 Glycosylation 26:38 N-linked Glycosylation 17:38 N-linked Glycosylation 17:38 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 3 4 used used 424:427 arg2 fragmentation 392:404 negative ion fragmentation 379:404 negative ion fragmentation of glycans 379:415 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 2 5 theme mass 289:292 arg1 spectrometry 294:305 ion mobility mass spectrometry 276:305 ion mobility mass spectrometry 276:305 Here, we describe how ion mobility mass spectrometry can be used to obtain spectra from virion samples. 26169737 1 6 theme Glycan 115:120 arg1 analysis 122:129 Glycan analysis 115:129 Glycan analysis of virion-derived glycoproteins 115:161 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 2 7 theme mobility 280:287 arg1 spectrometry 294:305 ion mobility mass spectrometry 276:305 ion mobility mass spectrometry 276:305 Here, we describe how ion mobility mass spectrometry can be used to obtain spectra from virion samples. 26169737 1 8 gly glycoprotein 205:216 arg1 glycoprotein 205:216 glycoprotein isolation 205:226 glycoprotein isolation 205:226 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 2 9 theme ion 276:278 arg1 spectrometry 294:305 ion mobility mass spectrometry 276:305 ion mobility mass spectrometry 276:305 Here, we describe how ion mobility mass spectrometry can be used to obtain spectra from virion samples. 26169737 3 10 theme negative 379:386 arg1 fragmentation 392:404 negative ion fragmentation 379:404 negative ion fragmentation of glycans 379:415 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 1 11 link virion-derived 134:147 arg1 glycoproteins 149:161 virion-derived glycoproteins 134:161 virion-derived glycoproteins 134:161 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 3 12 theme ion 388:390 arg1 fragmentation 392:404 negative ion fragmentation 379:404 negative ion fragmentation of glycans 379:415 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 0 13 theme Glycosylation 26:38 arg1 Determination 0:12 Determination 0:12 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility 0:112 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 1 14 theme glycoprotein 205:216 arg1 isolation 218:226 glycoprotein isolation 205:226 glycoprotein isolation 205:226 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 0 15 theme N-linked 17:24 arg1 Glycosylation 26:38 N-linked Glycosylation 17:38 N-linked Glycosylation 17:38 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 3 16 theme virion 461:466 arg1 glycans 468:474 virion glycans 461:474 virion glycans 461:474 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 0 17 theme Viral 43:47 arg1 Glycoproteins 49:61 Viral Glycoproteins 43:61 Viral Glycoproteins 43:61 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 3 18 theme structural 438:447 arg1 features 449:456 structural features 438:456 structural features of virion glycans 438:474 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 1 19 theme low 232:234 arg1 abundance 243:251 low sample abundance 232:251 low sample abundance 232:251 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 2 20 theme virion 342:347 arg1 samples 349:355 virion samples 342:355 virion samples 342:355 Here, we describe how ion mobility mass spectrometry can be used to obtain spectra from virion samples. 26169737 1 21 gly glycoproteins 149:161 arg1 glycoproteins 149:161 virion-derived glycoproteins 134:161 virion-derived glycoproteins 134:161 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 3 22 theme glycans 409:415 arg1 fragmentation 392:404 negative ion fragmentation 379:404 negative ion fragmentation of glycans 379:415 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 3 23 theme glycans 468:474 arg1 features 449:456 structural features 438:456 structural features of virion glycans 438:474 We also describe how negative ion fragmentation of glycans can be used to probe structural features of virion glycans. 26169737 1 24 theme sample 236:241 arg1 abundance 243:251 low sample abundance 232:251 low sample abundance 232:251 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 1 25 from difficulties 189:200 arg1 isolation 218:226 glycoprotein isolation 205:226 glycoprotein isolation 205:226 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 1 25 from difficulties 189:200 arg1 abundance 243:251 low sample abundance 232:251 low sample abundance 232:251 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 1 26 theme virion-derived 134:147 arg1 glycoproteins 149:161 virion-derived glycoproteins 134:161 virion-derived glycoproteins 134:161 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 2 27 used used 314:317 arg2 spectrometry 294:305 ion mobility mass spectrometry 276:305 ion mobility mass spectrometry 276:305 Here, we describe how ion mobility mass spectrometry can be used to obtain spectra from virion samples. 26169737 0 28 theme Ion 75:77 arg1 Spectrometry 84:95 Negative Ion Mass Spectrometry 66:95 Negative Ion Mass Spectrometry 66:95 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 26169737 1 29 theme glycoproteins 149:161 arg1 analysis 122:129 Glycan analysis 115:129 Glycan analysis of virion-derived glycoproteins 115:161 Glycan analysis of virion-derived glycoproteins is challenging due to the difficulties in glycoprotein isolation and low sample abundance. 26169737 0 30 theme Negative 66:73 arg1 Spectrometry 84:95 Negative Ion Mass Spectrometry 66:95 Negative Ion Mass Spectrometry 66:95 Determination of N-linked Glycosylation in Viral Glycoproteins by Negative Ion Mass Spectrometry and Ion Mobility. 24499412 5 0 theme zebrafish 1018:1026 arg1 embryogenesis 1028:1040 early zebrafish embryogenesis 1012:1040 early zebrafish embryogenesis 1012:1040 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 1 1 theme minute 154:159 arg1 scale 166:170 the minute time scale 150:170 the minute time scale 150:170 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 8 2 theme cleavage 1412:1419 arg1 period 1421:1426 the cleavage period 1408:1426 the cleavage period (0.75-2 hpf) 1408:1439 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 2 theme cleavage 1412:1419 arg1 hpf 1436:1438 0.75-2 hpf 1429:1438 0.75-2 hpf 1429:1438 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 4 3 theme cell-surface 816:827 arg1 glycans 829:835 newly synthesized cell-surface glycans 798:835 newly synthesized cell-surface glycans 798:835 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 3 4 theme living 636:641 arg1 systems 643:649 living systems 636:649 living systems 636:649 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 8 5 theme synthesized 1371:1381 arg1 glycans 1383:1389 newly synthesized glycans 1365:1389 newly synthesized glycans 1365:1389 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 1 6 theme time 161:164 arg1 scale 166:170 the minute time scale 150:170 the minute time scale 150:170 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 2 7 with azide 270:274 arg1 motif 309:313 an internal Cu(I)-chelating motif 281:313 an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis 281:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 5 8 from biosynthesis 973:984 arg1 embryogenesis 1028:1040 early zebrafish embryogenesis 1012:1040 early zebrafish embryogenesis 1012:1040 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 5 8 from biosynthesis 973:984 arg1 cells 999:1003 mammalian cells 989:1003 mammalian cells 989:1003 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 2 9 theme -chelating 298:307 arg1 motif 309:313 an internal Cu(I)-chelating motif 281:313 an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis 281:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 10 theme azide 270:274 arg1 discovery 254:262 our discovery 250:262 our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis 250:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 3 11 from efficiency 579:588 arg1 systems 643:649 living systems 636:649 living systems 636:649 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 5 12 theme mammalian 989:997 arg1 cells 999:1003 mammalian cells 989:1003 mammalian cells 989:1003 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 4 13 theme azide 705:709 arg1 combination 685:695 a combination 683:695 a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I) 683:772 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 7 14 located detected 1277:1284 arg1 embryos 1239:1245 zebrafish embryos 1229:1245 zebrafish embryos 1229:1245 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 7 14 located detected 1277:1284 arg2 glycans 1260:1266 the labeled glycans 1248:1266 the labeled glycans 1248:1266 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 7 14 located detected 1277:1284 arg2 stage 1311:1315 the two-cell stage 1298:1315 the two-cell stage 1298:1315 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 7 15 theme zebrafish 1229:1237 arg1 embryos 1239:1245 zebrafish embryos 1229:1245 zebrafish embryos 1229:1245 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 0 16 theme supersensitive 47:60 arg1 chemistry 68:76 supersensitive click chemistry 47:76 supersensitive click chemistry 47:76 Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. 24499412 3 17 theme apparent 659:666 arg1 toxicity 668:675 no apparent toxicity 656:675 no apparent toxicity 656:675 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 8 18 theme bioorthogonal 1466:1478 arg1 chemistry 1480:1488 bioorthogonal chemistry 1466:1488 bioorthogonal chemistry 1466:1488 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 4 19 theme BTTPS 715:719 arg1 combination 685:695 a combination 683:695 a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I) 683:772 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 8 20 located detected 1396:1403 arg2 glycans 1383:1389 newly synthesized glycans 1365:1389 newly synthesized glycans 1365:1389 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 20 located detected 1396:1403 arg1 period 1421:1426 the cleavage period 1408:1426 the cleavage period (0.75-2 hpf) 1408:1439 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 20 located detected 1396:1403 arg1 model 1454:1458 an animal model 1444:1458 an animal model using bioorthogonal chemistry 1444:1488 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 20 located detected 1396:1403 arg1 hpf 1436:1438 0.75-2 hpf 1429:1438 0.75-2 hpf 1429:1438 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 2 21 theme -catalyzed 377:386 arg1 cycloaddition 401:413 Cu(I)-catalyzed azide-alkyne cycloaddition 372:413 Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date 372:429 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 21 theme -catalyzed 377:386 arg1 CuAAC 416:420 CuAAC 416:420 CuAAC 416:420 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 22 theme internal 284:291 arg1 motif 309:313 an internal Cu(I)-chelating motif 281:313 an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis 281:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 1 23 theme simple 173:178 arg1 methods 200:206 simple and fast analytical methods 173:206 simple and fast analytical methods 173:206 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 6 24 theme building 1141:1148 arg1 block 1150:1154 a monosaccharide building block 1124:1154 a monosaccharide building block 1124:1154 In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. 24499412 6 25 theme mammalian 1051:1059 arg1 cells 1061:1065 live mammalian cells 1046:1065 live mammalian cells 1046:1065 In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. 24499412 7 26 theme labeled 1252:1258 arg1 stage 1311:1315 the two-cell stage 1298:1315 the two-cell stage 1298:1315 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 7 26 theme labeled 1252:1258 arg1 glycans 1260:1266 the labeled glycans 1248:1266 the labeled glycans 1248:1266 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 6 27 theme live 1046:1049 arg1 cells 1061:1065 live mammalian cells 1046:1065 live mammalian cells 1046:1065 In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. 24499412 6 28 theme monosaccharide 1126:1139 arg1 block 1150:1154 a monosaccharide building block 1124:1154 a monosaccharide building block 1124:1154 In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. 24499412 1 29 theme fast 184:187 arg1 methods 200:206 simple and fast analytical methods 173:206 simple and fast analytical methods 173:206 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 2 30 theme dynamic 473:479 arg1 process 481:487 the dynamic process 469:487 the dynamic process of glycan biosynthesis 469:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 5 31 theme dynamic 958:964 arg1 biosynthesis 973:984 the dynamic glycan biosynthesis 954:984 the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis 954:1040 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 1 32 theme biological 106:115 arg1 processes 117:125 biological processes 106:125 biological processes that take place within the minute time scale 106:170 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 1 33 theme analytical 189:198 arg1 methods 200:206 simple and fast analytical methods 173:206 simple and fast analytical methods 173:206 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 4 34 theme metabolic 881:889 arg1 precursor 891:899 a metabolic precursor 879:899 a metabolic precursor 879:899 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 4 35 theme flow 840:843 arg1 cytometry 845:853 flow cytometry 840:853 flow cytometry using as low as 1 nM of a metabolic precursor 840:899 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 5 36 theme glycan 966:971 arg1 biosynthesis 973:984 the dynamic glycan biosynthesis 954:984 the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis 954:1040 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 4 37 theme amine-based 745:755 arg1 ligand 757:762 a tris(triazolylmethyl)amine-based ligand 722:762 a tris(triazolylmethyl)amine-based ligand for Cu(I) 722:772 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 4 37 theme amine-based 745:755 arg1 azide 705:709 azide 705:709 azide 705:709 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 2 38 theme protocol 359:366 arg1 application 440:450 its application 436:450 its application toward following the dynamic process of glycan biosynthesis 436:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 38 theme protocol 359:366 arg1 development 332:342 the development 328:342 the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date 328:429 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 4 39 theme precursor 891:899 arg1 nM 873:874 1 nM 871:874 1 nM of a metabolic precursor 871:899 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 8 40 theme first 1349:1353 arg1 this 1336:1339 this 1336:1339 this 1336:1339 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 40 theme first 1349:1353 arg1 time 1355:1358 the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry 1345:1488 the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry 1345:1488 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 5 41 theme supersensitive 907:920 arg1 chemistry 922:930 This supersensitive chemistry 902:930 This supersensitive chemistry 902:930 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 2 42 theme fastest 351:357 arg1 protocol 359:366 the fastest protocol 347:366 the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date 347:429 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 0 43 theme click 62:66 arg1 chemistry 68:76 supersensitive click chemistry 47:76 supersensitive click chemistry 47:76 Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. 24499412 8 44 theme animal 1447:1452 arg1 model 1454:1458 an animal model 1444:1458 an animal model using bioorthogonal chemistry 1444:1488 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 45 theme 0.75-2 1429:1434 arg1 period 1421:1426 the cleavage period 1408:1426 the cleavage period (0.75-2 hpf) 1408:1439 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 8 45 theme 0.75-2 1429:1434 arg1 hpf 1436:1438 0.75-2 hpf 1429:1438 0.75-2 hpf 1429:1438 To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry. 24499412 4 46 theme synthesized 804:814 arg1 glycans 829:835 newly synthesized cell-surface glycans 798:835 newly synthesized cell-surface glycans 798:835 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 1 47 theme processes 117:125 arg1 kinetics 94:101 the kinetics 90:101 the kinetics of biological processes that take place within the minute time scale 90:170 To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. 24499412 2 48 theme azide-alkyne 388:399 arg1 cycloaddition 401:413 Cu(I)-catalyzed azide-alkyne cycloaddition 372:413 Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date 372:429 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 48 theme azide-alkyne 388:399 arg1 CuAAC 416:420 CuAAC 416:420 CuAAC 416:420 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 49 theme biosynthesis 499:510 arg1 process 481:487 the dynamic process 469:487 the dynamic process of glycan biosynthesis 469:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 3 50 theme 20-38-fold 622:631 arg1 efficiency 579:588 the efficiency 575:588 the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems 575:649 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 4 51 with able 783:786 arg1 combination 685:695 a combination 683:695 a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I) 683:772 With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. 24499412 2 52 theme glycan 492:497 arg1 biosynthesis 499:510 glycan biosynthesis 492:510 glycan biosynthesis 492:510 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 7 53 theme two-cell 1302:1309 arg1 stage 1311:1315 the two-cell stage 1298:1315 the two-cell stage 1298:1315 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 7 53 theme two-cell 1302:1309 arg1 glycans 1260:1266 the labeled glycans 1248:1266 the labeled glycans 1248:1266 In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. 24499412 6 54 theme cell-surface 1196:1207 arg1 glycoconjugates 1209:1223 cell-surface glycoconjugates 1196:1223 cell-surface glycoconjugates 1196:1223 In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. 24499412 3 55 theme ligand-accelerated 597:614 arg1 20-38-fold 622:631 the ligand-accelerated CuAAC 20-38-fold 593:631 the ligand-accelerated CuAAC 20-38-fold 593:631 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 2 56 attach present 242:248 arg2 we 239:240 we 239:240 we 239:240 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 2 56 attach present 242:248 arg1 article 230:236 this article 225:236 this article 225:236 In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. 24499412 3 57 theme CuAAC 616:620 arg1 20-38-fold 622:631 the ligand-accelerated CuAAC 20-38-fold 593:631 the ligand-accelerated CuAAC 20-38-fold 593:631 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 3 58 theme electron-donating 535:551 arg1 azide 561:565 an electron-donating picolyl azide 532:565 an electron-donating picolyl azide 532:565 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 5 59 theme early 1012:1016 arg1 embryogenesis 1028:1040 early zebrafish embryogenesis 1012:1040 early zebrafish embryogenesis 1012:1040 This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. 24499412 3 60 theme picolyl 553:559 arg1 azide 561:565 an electron-donating picolyl azide 532:565 an electron-donating picolyl azide 532:565 We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. 24499412 0 61 theme dynamic 11:17 arg1 glycosylation 19:31 dynamic glycosylation 11:31 dynamic glycosylation 11:31 Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. 24591635 6 0 theme lectin 760:765 arg1 microarrays 767:777 lectin microarrays 760:777 lectin microarrays 760:777 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 1 1 theme biological 158:167 arg1 milieu 169:174 the biological milieu 154:174 the biological milieu 154:174 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 8 2 theme regulatory 1145:1154 arg1 networks 1156:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 9 3 theme miRNA-mediated 1448:1461 arg1 control 1463:1469 miRNA-mediated control 1448:1469 miRNA-mediated control of cell phenotype and the glycome 1448:1503 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 7 4 from genes 995:999 arg1 glycogenes 1034:1043 glycogenes 1034:1043 glycogenes 1034:1043 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 7 4 from genes 995:999 arg1 pathways 1024:1031 glycan biosynthetic pathways 1004:1031 glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures 1004:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 2 5 theme proteins 356:363 arg1 glycosidases 401:412 glycosidases 401:412 glycosidases 401:412 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 2 5 theme proteins 356:363 arg1 proteins 356:363 proteins 356:363 proteins 356:363 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 2 5 theme proteins 356:363 arg1 glycosyltransferases 376:395 glycosyltransferases 376:395 glycosyltransferases 376:395 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 2 5 theme proteins 356:363 arg1 repertoire 342:351 a repertoire 340:351 a repertoire of proteins, including glycosyltransferases and glycosidases 340:412 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 8 6 dep three 1104:1108 arg1 fucose 1180:1185 fucose 1180:1185 fucose 1180:1185 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 8 6 dep three 1104:1108 arg1 mannose 1171:1177 high mannose 1166:1177 high mannose 1166:1177 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 8 6 dep three 1104:1108 arg1 β-GalNAc 1201:1208 terminal β-GalNAc 1192:1208 terminal β-GalNAc 1192:1208 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 5 7 theme unique 634:639 arg1 approach 655:662 a unique systems-based approach 632:662 a unique systems-based approach to identify connections between miRNA and the glycome 632:716 Herein we take a unique systems-based approach to identify connections between miRNA and the glycome. 24591635 2 8 theme coordinated 316:326 arg1 activity 328:335 the coordinated activity 312:335 the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases 312:412 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 7 9 theme glycan 1004:1009 arg1 glycogenes 1034:1043 glycogenes 1034:1043 glycogenes 1034:1043 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 7 9 theme glycan 1004:1009 arg1 pathways 1024:1031 glycan biosynthetic pathways 1004:1031 glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures 1004:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 9 10 theme critical 1338:1345 arg1 nodes 1347:1351 critical nodes 1338:1351 critical nodes 1338:1351 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 9 11 theme global 1360:1365 arg1 network 1381:1387 the global glycosylation network 1356:1387 the global glycosylation network accessible to miRNA regulation 1356:1418 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 5 12 theme systems-based 641:653 arg1 approach 655:662 a unique systems-based approach 632:662 a unique systems-based approach to identify connections between miRNA and the glycome 632:716 Herein we take a unique systems-based approach to identify connections between miRNA and the glycome. 24591635 9 13 theme cell 1474:1477 arg1 phenotype 1479:1487 cell phenotype 1474:1487 cell phenotype 1474:1487 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 8 14 theme traditional 1277:1287 arg1 methods 1303:1309 traditional bioinformatic methods 1277:1309 traditional bioinformatic methods 1277:1309 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 3 15 theme biosynthetic 486:497 arg1 process 499:505 this complex biosynthetic process 473:505 this complex biosynthetic process 473:505 Little is known about the regulatory networks controlling this complex biosynthetic process. 24591635 0 16 theme glycome 52:58 arg1 regulation 28:37 Mapping posttranscriptional regulation 0:37 Mapping posttranscriptional regulation of the human glycome 0:58 Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode. 24591635 4 17 from role 532:535 arg1 regulation 565:574 the regulation 561:574 the regulation of specific glycan biosynthetic enzymes 561:614 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 2 18 theme structures 292:301 arg1 Assembly 258:265 Assembly 258:265 Assembly of discrete carbohydrate structures 258:301 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 6 19 theme analysis 741:748 arg1 platform 750:757 our glycomic analysis platform 728:757 our glycomic analysis platform 728:757 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 0 20 theme posttranscriptional 8:26 arg1 regulation 28:37 Mapping posttranscriptional regulation 0:37 Mapping posttranscriptional regulation of the human glycome 0:58 Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode. 24591635 6 21 theme flow 910:913 arg1 output 880:885 a direct output 871:885 a direct output of genomic information flow 871:913 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 6 22 theme cell 831:834 arg1 panel 836:840 the NCI-60 cell panel 820:840 the NCI-60 cell panel 820:840 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 2 23 theme repertoire 342:351 arg1 activity 328:335 the coordinated activity 312:335 the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases 312:412 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 6 24 theme glycomic 732:739 arg1 platform 750:757 our glycomic analysis platform 728:757 our glycomic analysis platform 728:757 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 1 25 theme diverse 187:193 arg1 processes 195:203 diverse processes 187:203 diverse processes 187:203 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 0 26 theme Mapping 0:6 arg1 regulation 28:37 Mapping posttranscriptional regulation 0:37 Mapping posttranscriptional regulation of the human glycome 0:58 Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode. 24591635 4 27 theme glycan 588:593 arg1 enzymes 608:614 specific glycan biosynthetic enzymes 579:614 specific glycan biosynthetic enzymes 579:614 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 6 28 theme NCI-60 824:829 arg1 panel 836:840 the NCI-60 cell panel 820:840 the NCI-60 cell panel 820:840 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 1 29 theme Cell 102:105 arg1 glycans 115:121 Cell surface glycans 102:121 Cell surface glycans 102:121 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 3 30 theme complex 478:484 arg1 process 499:505 this complex biosynthetic process 473:505 this complex biosynthetic process 473:505 Little is known about the regulatory networks controlling this complex biosynthetic process. 24591635 4 31 theme specific 579:586 arg1 enzymes 608:614 specific glycan biosynthetic enzymes 579:614 specific glycan biosynthetic enzymes 579:614 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 1 32 theme surface 107:113 arg1 glycans 115:121 Cell surface glycans 102:121 Cell surface glycans 102:121 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 8 33 theme miRNA 1223:1227 arg1 regulation 1229:1238 miRNA regulation 1223:1238 miRNA regulation that would not have been observed by traditional bioinformatic methods 1223:1309 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 4 34 theme enzymes 608:614 arg1 regulation 565:574 the regulation 561:574 the regulation of specific glycan biosynthetic enzymes 561:614 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 4 35 theme Recent 508:513 arg1 work 515:518 Recent work 508:518 Recent work 508:518 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 8 36 theme bioinformatic 1289:1301 arg1 methods 1303:1309 traditional bioinformatic methods 1277:1309 traditional bioinformatic methods 1277:1309 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 0 37 theme human 46:50 arg1 glycome 52:58 the human glycome 42:58 the human glycome 42:58 Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode. 24591635 4 38 theme biosynthetic 595:606 arg1 enzymes 608:614 specific glycan biosynthetic enzymes 579:614 specific glycan biosynthetic enzymes 579:614 Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. 24591635 8 39 theme predicted 1119:1127 arg1 networks 1156:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 6 40 theme genomic 890:896 arg1 flow 910:913 genomic information flow 890:913 genomic information flow 890:913 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 9 41 theme glycome 1497:1503 arg1 control 1463:1469 miRNA-mediated control 1448:1469 miRNA-mediated control of cell phenotype and the glycome 1448:1503 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 1 42 theme inflammatory 214:225 arg1 cascade 227:233 the inflammatory cascade 210:233 the inflammatory cascade to cellular migration 210:255 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 9 43 theme phenotype 1479:1487 arg1 control 1463:1469 miRNA-mediated control 1448:1469 miRNA-mediated control of cell phenotype and the glycome 1448:1503 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 8 44 theme terminal 1192:1199 arg1 β-GalNAc 1201:1208 terminal β-GalNAc 1192:1208 terminal β-GalNAc 1192:1208 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 9 45 theme miRNA 1403:1407 arg1 regulation 1409:1418 miRNA regulation 1403:1418 miRNA regulation 1403:1418 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 7 46 theme observed 1064:1071 arg1 structures 1080:1089 the observed glycan structures 1060:1089 the observed glycan structures 1060:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 9 47 theme glycosylation 1367:1379 arg1 network 1381:1387 the global glycosylation network 1356:1387 the global glycosylation network accessible to miRNA regulation 1356:1418 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 6 48 theme information 898:908 arg1 flow 910:913 genomic information flow 890:913 genomic information flow 890:913 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 8 49 theme miRNA/glycogene 1129:1143 arg1 networks 1156:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 7 50 theme miRNA 973:977 arg1 regulators 979:988 miRNA regulators 973:988 miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures 973:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 3 51 theme regulatory 441:450 arg1 networks 452:459 the regulatory networks 437:459 the regulatory networks controlling this complex biosynthetic process 437:505 Little is known about the regulatory networks controlling this complex biosynthetic process. 24591635 7 52 theme glycan 1073:1078 arg1 structures 1080:1089 the observed glycan structures 1060:1089 the observed glycan structures 1060:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 6 53 theme direct 873:878 arg1 output 880:885 a direct output 871:885 a direct output of genomic information flow 871:913 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 6 54 theme glycosylation 792:804 arg1 signatures 806:815 glycosylation signatures 792:815 glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow 792:913 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 2 55 theme carbohydrate 279:290 arg1 structures 292:301 discrete carbohydrate structures 270:301 discrete carbohydrate structures 270:301 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 1 56 theme critical 130:137 arg1 interface 139:147 a critical interface 128:147 a critical interface 128:147 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 7 57 theme miRNA 954:958 arg1 data 960:963 miRNA data 954:963 miRNA data 954:963 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 8 58 theme networks 1156:1163 arg1 three 1104:1108 three 1104:1108 three 1104:1108 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 8 58 theme networks 1156:1163 arg1 networks 1156:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 these predicted miRNA/glycogene regulatory networks 1113:1163 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 24591635 2 59 theme discrete 270:277 arg1 structures 292:301 discrete carbohydrate structures 270:301 discrete carbohydrate structures 270:301 Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. 24591635 7 60 theme biosynthetic 1011:1022 arg1 glycogenes 1034:1043 glycogenes 1034:1043 glycogenes 1034:1043 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 7 60 theme biosynthetic 1011:1022 arg1 pathways 1024:1031 glycan biosynthetic pathways 1004:1031 glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures 1004:1089 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 6 61 from signatures 806:815 arg1 panel 836:840 the NCI-60 cell panel 820:840 the NCI-60 cell panel 820:840 By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. 24591635 7 62 theme glycomic 932:939 arg1 dataset 941:947 our glycomic dataset 928:947 our glycomic dataset 928:947 Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. 24591635 1 63 theme cellular 238:245 arg1 migration 247:255 cellular migration 238:255 cellular migration 238:255 Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. 24591635 9 64 theme accessible 1389:1398 arg1 network 1381:1387 the global glycosylation network 1356:1387 the global glycosylation network accessible to miRNA regulation 1356:1418 Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome. 24591635 8 65 theme high 1166:1169 arg1 mannose 1171:1177 high mannose 1166:1177 high mannose 1166:1177 We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. 25924827 5 0 theme studied 684:690 arg1 protein 692:698 the studied protein 680:698 the studied protein 680:698 No isotope labeling is required, nor does the molecular weight of the studied protein limit the application. 25924827 7 1 attach derived 886:892 arg2 proteins 877:884 proteins 877:884 proteins derived from native organisms 877:914 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 7 1 attach derived 886:892 arg1 organisms 906:914 native organisms 899:914 native organisms 899:914 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 5 2 theme molecular 660:668 arg1 weight 670:675 the molecular weight 656:675 the molecular weight of the studied protein 656:698 No isotope labeling is required, nor does the molecular weight of the studied protein limit the application. 25924827 7 3 theme therapeutic 975:985 arg1 proteins 987:994 biotechnologically produced therapeutic proteins 947:994 biotechnologically produced therapeutic proteins 947:994 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 4 4 theme PTMs 596:599 arg1 identification 578:591 identification 578:591 identification 578:591 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 4 4 theme PTMs 596:599 arg1 detection 564:572 detection 564:572 detection 564:572 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 3 5 theme sample 356:361 arg1 preparations 363:374 complicated sample preparations 344:374 complicated sample preparations 344:374 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 4 6 dep detection 564:572 arg1 the 560:562 the 560:562 the 560:562 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 5 7 theme isotope 617:623 arg1 labeling 625:632 No isotope labeling 614:632 No isotope labeling 614:632 No isotope labeling is required, nor does the molecular weight of the studied protein limit the application. 25924827 7 8 theme proteins 877:884 arg1 diagnostics 862:872 diagnostics 862:872 diagnostics of proteins derived from native organisms 862:914 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 4 9 from identification 578:591 arg1 proteins 604:611 proteins 604:611 proteins 604:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 4 10 theme direct 505:510 arg1 method 549:554 a direct, robust, and simple NMR spectroscopy method 503:554 a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins 503:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 6 11 theme sophisticated 795:807 arg1 preparation 816:826 sophisticated sample preparation 795:826 sophisticated sample preparation 795:826 The method can directly detect modifications on intact proteins without sophisticated sample preparation. 25924827 4 12 from detection 564:572 arg1 proteins 604:611 proteins 604:611 proteins 604:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 3 13 dep methods 312:318 arg1 analyze 323:329 analyze 323:329 to analyze PTMs 320:334 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 3 14 theme certain 400:406 arg1 structure 481:489 thus chemical structure 467:489 thus chemical structure 467:489 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 3 14 theme certain 400:406 arg1 modifications 408:420 certain modifications 400:420 certain modifications 400:420 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 3 14 theme certain 400:406 arg1 inability 427:435 the inability to identify linkage types 423:461 the inability to identify linkage types 423:461 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 7 15 theme produced 966:973 arg1 proteins 987:994 biotechnologically produced therapeutic proteins 947:994 biotechnologically produced therapeutic proteins 947:994 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 0 16 theme Posttranslational 0:16 arg1 modifications 18:30 Posttranslational modifications 0:30 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 0:109 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 25924827 3 17 theme linkage 449:455 arg1 types 457:461 linkage types 449:461 linkage types 449:461 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 0 18 theme intact 35:40 arg1 proteins 42:49 intact proteins 35:49 intact proteins detected by NMR spectroscopy 35:78 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 25924827 5 19 theme protein 692:698 arg1 weight 670:675 the molecular weight 656:675 the molecular weight of the studied protein 656:698 No isotope labeling is required, nor does the molecular weight of the studied protein limit the application. 25924827 6 20 theme sample 809:814 arg1 preparation 816:826 sophisticated sample preparation 795:826 sophisticated sample preparation 795:826 The method can directly detect modifications on intact proteins without sophisticated sample preparation. 25924827 1 21 theme Posttranslational 111:127 arg1 part 166:169 an integral part 154:169 an integral part of the majority of proteins 154:197 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 1 21 theme Posttranslational 111:127 arg1 PTMs 144:147 PTMs 144:147 PTMs 144:147 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 1 21 theme Posttranslational 111:127 arg1 modifications 129:141 Posttranslational modifications 111:141 Posttranslational modifications (PTMs) 111:148 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 7 22 theme proteins 987:994 arg1 control 936:942 the quality control 924:942 the quality control of biotechnologically produced therapeutic proteins 924:994 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 1 23 theme proteins 190:197 arg1 majority 178:185 the majority 174:185 the majority of proteins 174:197 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 0 24 theme proteins 42:49 arg1 modifications 18:30 Posttranslational modifications 0:30 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 0:109 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 25924827 4 25 theme NMR 532:534 arg1 spectroscopy 536:547 NMR spectroscopy 532:547 a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins 503:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 2 26 theme function 238:245 arg1 challenging 267:277 challenging 267:277 challenging 267:277 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 2 26 theme function 238:245 arg1 characterization 204:219 The characterization 200:219 The characterization of structure and function of PTMs 200:253 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 3 27 theme chemical 472:479 arg1 modifications 408:420 certain modifications 400:420 certain modifications 400:420 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 3 27 theme chemical 472:479 arg1 structure 481:489 thus chemical structure 467:489 thus chemical structure 467:489 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 0 28 theme NMR 63:65 arg1 spectroscopy 67:78 NMR spectroscopy 63:78 NMR spectroscopy 63:78 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 25924827 4 29 theme simple 525:530 arg1 method 549:554 a direct, robust, and simple NMR spectroscopy method 503:554 a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins 503:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 6 30 theme intact 771:776 arg1 proteins 778:785 intact proteins 771:785 intact proteins without sophisticated sample preparation 771:826 The method can directly detect modifications on intact proteins without sophisticated sample preparation. 25924827 2 31 theme structure 224:232 arg1 challenging 267:277 challenging 267:277 challenging 267:277 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 2 31 theme structure 224:232 arg1 characterization 204:219 The characterization 200:219 The characterization of structure and function of PTMs 200:253 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 4 32 theme spectroscopy 536:547 arg1 method 549:554 a direct, robust, and simple NMR spectroscopy method 503:554 a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins 503:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 7 33 theme quality 928:934 arg1 control 936:942 the quality control 924:942 the quality control of biotechnologically produced therapeutic proteins 924:994 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 25924827 3 34 theme Existing 303:310 arg1 methods 312:318 Existing methods 303:318 Existing methods to analyze PTMs 303:334 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 6 35 from modifications 754:766 arg1 proteins 778:785 intact proteins 771:785 intact proteins without sophisticated sample preparation 771:826 The method can directly detect modifications on intact proteins without sophisticated sample preparation. 25924827 0 36 dep modifications 18:30 arg1 application 81:91 application 81:91 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 0:109 Posttranslational modifications of intact proteins detected by NMR spectroscopy: application to glycosylation. 25924827 1 37 theme majority 178:185 arg1 part 166:169 an integral part 154:169 an integral part of the majority of proteins 154:197 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 1 37 theme majority 178:185 arg1 modifications 129:141 Posttranslational modifications 111:141 Posttranslational modifications (PTMs) 111:148 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 3 38 theme complicated 344:354 arg1 preparations 363:374 complicated sample preparations 344:374 complicated sample preparations 344:374 Existing methods to analyze PTMs require complicated sample preparations and suffer from missing certain modifications, the inability to identify linkage types and thus chemical structure. 25924827 4 39 theme robust 513:518 arg1 method 549:554 a direct, robust, and simple NMR spectroscopy method 503:554 a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins 503:611 We present a direct, robust, and simple NMR spectroscopy method for the detection and identification of PTMs in proteins. 25924827 2 40 theme PTMs 250:253 arg1 function 238:245 function 238:245 function 238:245 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 2 40 theme PTMs 250:253 arg1 structure 224:232 structure 224:232 structure 224:232 The characterization of structure and function of PTMs can be very challenging especially for glycans. 25924827 1 41 theme integral 157:164 arg1 part 166:169 an integral part 154:169 an integral part of the majority of proteins 154:197 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 1 41 theme integral 157:164 arg1 modifications 129:141 Posttranslational modifications 111:141 Posttranslational modifications (PTMs) 111:148 Posttranslational modifications (PTMs) are an integral part of the majority of proteins. 25924827 7 42 theme native 899:904 arg1 organisms 906:914 native organisms 899:914 native organisms 899:914 This approach is well suited for diagnostics of proteins derived from native organisms and for the quality control of biotechnologically produced therapeutic proteins. 27810711 0 0 theme glycopeptides 69:81 arg1 synthesis 37:45 chemical synthesis 28:45 chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains 28:113 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 27810711 3 1 gly glycopeptide 401:412 arg2 glycopeptide 401:412 syndecan-3 glycopeptide 390:412 syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 390:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 1 gly glycopeptide 401:412 arg2 53-62 415:419 53-62 415:419 53-62 415:419 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 6 2 theme groups 1049:1054 arg1 coupling 972:979 coupling 972:979 coupling of the glycan-bearing fragments 972:1011 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 2 theme groups 1049:1054 arg1 cleavage 1017:1024 cleavage 1017:1024 cleavage of the acyl protecting groups 1017:1054 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 4 3 theme amino 577:581 arg1 cassettes 588:596 two different octasaccharide amino acid cassettes 548:596 two different octasaccharide amino acid cassettes 548:596 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 5 4 theme peptide 733:739 arg1 elongation 741:750 peptide elongation 733:750 peptide elongation 733:750 The glycopeptides presented many obstacles for post-glycosylation manipulation, peptide elongation, and deprotection. 27810711 1 5 theme many 153:156 arg1 events 169:174 many biological events 153:174 many biological events 153:174 Proteoglycans play critical roles in many biological events. 27810711 4 6 theme octasaccharide 562:575 arg1 cassettes 588:596 two different octasaccharide amino acid cassettes 548:596 two different octasaccharide amino acid cassettes 548:596 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 2 7 theme glycan 296:301 arg1 chains 303:308 well-defined glycan chains 283:308 well-defined glycan chains 283:308 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 5 8 theme many 681:684 arg1 obstacles 686:694 many obstacles 681:694 many obstacles for post-glycosylation manipulation, peptide elongation, and deprotection 681:768 The glycopeptides presented many obstacles for post-glycosylation manipulation, peptide elongation, and deprotection. 27810711 2 9 theme well-defined 283:294 arg1 chains 303:308 well-defined glycan chains 283:308 well-defined glycan chains 283:308 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 4 10 theme acid 583:586 arg1 cassettes 588:596 two different octasaccharide amino acid cassettes 548:596 two different octasaccharide amino acid cassettes 548:596 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 2 11 gly glycopeptides 261:273 arg2 glycopeptides 261:273 glycopeptides 261:273 glycopeptides bearing well-defined glycan chains 261:308 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 1 12 theme biological 158:167 arg1 events 169:174 many biological events 153:174 many biological events 153:174 Proteoglycans play critical roles in many biological events. 27810711 0 13 theme heparan 92:98 arg1 chains 108:113 two heparan sulfate chains 88:113 two heparan sulfate chains 88:113 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 27810711 6 14 theme successful 826:835 arg1 strategy 837:844 a successful strategy 824:844 a successful strategy 824:844 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 0 15 gly glycopeptides 69:81 arg2 glycopeptides 69:81 syndecan-3 (53-62) glycopeptides 50:81 syndecan-3 (53-62) glycopeptides 50:81 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 27810711 0 16 theme sulfate 100:106 arg1 chains 108:113 two heparan sulfate chains 88:113 two heparan sulfate chains 88:113 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 27810711 4 17 gly glycopeptides 638:650 arg2 glycopeptides 638:650 syndecan-3 glycopeptides 627:650 syndecan-3 glycopeptides 627:650 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 6 18 theme protecting 1038:1047 arg1 groups 1049:1054 the acyl protecting groups 1029:1054 the acyl protecting groups 1029:1054 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 3 19 theme full 357:360 arg1 account 362:368 the full account 353:368 the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 353:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 6 20 theme acyl 1033:1036 arg1 groups 1049:1054 the acyl protecting groups 1029:1054 the acyl protecting groups 1029:1054 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 21 theme sequences 813:821 arg1 screening 781:789 screening 781:789 screening of multiple synthetic sequences 781:821 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 3 22 theme heparan 447:453 arg1 chains 463:468 two different heparan sulfate chains 433:468 two different heparan sulfate chains 433:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 4 23 theme different 552:560 arg1 cassettes 588:596 two different octasaccharide amino acid cassettes 548:596 two different octasaccharide amino acid cassettes 548:596 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 4 24 used utilized 610:617 arg2 cassettes 588:596 two different octasaccharide amino acid cassettes 548:596 two different octasaccharide amino acid cassettes 548:596 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 6 25 theme containing 928:937 arg1 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 3 26 theme sulfate 455:461 arg1 chains 463:468 two different heparan sulfate chains 433:468 two different heparan sulfate chains 433:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 27 theme syndecan-3 390:399 arg1 53-62 415:419 53-62 415:419 53-62 415:419 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 27 theme syndecan-3 390:399 arg1 glycopeptide 401:412 syndecan-3 glycopeptide 390:412 syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 390:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 6 28 theme chain 922:926 arg1 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 0 29 theme chemical 28:35 arg1 synthesis 37:45 chemical synthesis 28:45 chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains 28:113 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 27810711 4 30 theme 3+2+3 509:513 arg1 approach 515:522 a convergent 3+2+3 approach 496:522 a convergent 3+2+3 approach 496:522 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 3 31 theme glycopeptide 401:412 arg1 synthesis 377:385 the synthesis 373:385 the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 373:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 6 32 theme glycan 915:920 arg1 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 33 theme synthetic 803:811 arg1 sequences 813:821 multiple synthetic sequences 794:821 multiple synthetic sequences 794:821 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 3 34 theme different 437:445 arg1 chains 463:468 two different heparan sulfate chains 433:468 two different heparan sulfate chains 433:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 35 theme synthesis 377:385 arg1 account 362:368 the full account 353:368 the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 353:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 4 36 theme convergent 498:507 arg1 approach 515:522 a convergent 3+2+3 approach 496:522 a convergent 3+2+3 approach 496:522 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 6 37 theme single 908:913 arg1 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 5 38 gly glycopeptides 657:669 arg2 glycopeptides 657:669 The glycopeptides 653:669 The glycopeptides 653:669 The glycopeptides presented many obstacles for post-glycosylation manipulation, peptide elongation, and deprotection. 27810711 6 39 theme fragments 1003:1011 arg1 coupling 972:979 coupling 972:979 coupling of the glycan-bearing fragments 972:1011 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 39 theme fragments 1003:1011 arg1 cleavage 1017:1024 cleavage 1017:1024 cleavage of the acyl protecting groups 1017:1054 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 40 theme deprotected 896:906 arg1 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 6 41 theme glycan-bearing 988:1001 arg1 fragments 1003:1011 the glycan-bearing fragments 984:1011 the glycan-bearing fragments 984:1011 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 5 42 theme post-glycosylation 700:717 arg1 manipulation 719:730 post-glycosylation manipulation 700:730 post-glycosylation manipulation 700:730 The glycopeptides presented many obstacles for post-glycosylation manipulation, peptide elongation, and deprotection. 27810711 2 43 theme structural 190:199 arg1 complexities 201:212 their structural complexities 184:212 their structural complexities 184:212 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 6 44 theme multiple 794:801 arg1 sequences 813:821 multiple synthetic sequences 794:821 multiple synthetic sequences 794:821 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 2 45 theme glycopeptides 261:273 arg1 class 252:256 this class 247:256 this class of glycopeptides bearing well-defined glycan chains 247:308 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 3 46 contain containing 422:431 arg2 chains 463:468 two different heparan sulfate chains 433:468 two different heparan sulfate chains 433:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 46 contain containing 422:431 arg1 53-62 415:419 53-62 415:419 53-62 415:419 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 3 46 contain containing 422:431 arg1 glycopeptide 401:412 syndecan-3 glycopeptide 390:412 syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains 390:468 In this work, we give the full account of the synthesis of syndecan-3 glycopeptide (53-62) containing two different heparan sulfate chains. 27810711 2 47 theme class 252:256 arg1 synthesis 234:242 synthesis 234:242 synthesis of this class of glycopeptides bearing well-defined glycan chains 234:308 Due to their structural complexities, strategies towards synthesis of this class of glycopeptides bearing well-defined glycan chains are urgently needed. 27810711 6 48 gly glycopeptides 939:951 arg2 glycopeptides 939:951 partially deprotected single glycan chain containing glycopeptides 886:951 partially deprotected single glycan chain containing glycopeptides 886:951 Following screening of multiple synthetic sequences, a successful strategy was finally established by constructing partially deprotected single glycan chain containing glycopeptides first, followed by coupling of the glycan-bearing fragments and cleavage of the acyl protecting groups. 27810711 4 49 theme syndecan-3 627:636 arg1 glycopeptides 638:650 syndecan-3 glycopeptides 627:650 syndecan-3 glycopeptides 627:650 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 1 50 theme critical 135:142 arg1 roles 144:148 critical roles 135:148 critical roles 135:148 Proteoglycans play critical roles in many biological events. 27810711 4 51 theme glycans 487:493 arg1 assembly 475:482 assembly 475:482 assembly of glycans 475:493 For assembly of glycans, a convergent 3+2+3 approach was developed producing two different octasaccharide amino acid cassettes, which were utilized towards syndecan-3 glycopeptides. 27810711 0 52 with synthesis 37:45 arg1 chains 108:113 two heparan sulfate chains 88:113 two heparan sulfate chains 88:113 Obstacles and solutions for chemical synthesis of syndecan-3 (53-62) glycopeptides with two heparan sulfate chains. 26650028 5 0 theme reaction 808:815 arg1 conditions 817:826 rigorous reaction conditions 799:826 rigorous reaction conditions 799:826 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 6 1 theme synthetic 956:964 arg1 efficiency 966:975 synthetic efficiency 956:975 synthetic efficiency 956:975 In addition, common intermediates and a series of one-pot reactions were employed to enhance synthetic efficiency, improving the yield considerably. 26650028 2 2 from blocks 418:423 arg1 yield 363:367 3.5% overall yield 350:367 3.5% overall yield from well-functionalized monosaccharide building blocks 350:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 4 3 theme functional-group 722:737 arg1 modifications 739:751 oligosaccharide stage functional-group modifications 700:751 oligosaccharide stage functional-group modifications 700:751 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 3 4 from step 530:533 arg1 blocks 585:590 the monosaccharide building blocks 557:590 the monosaccharide building blocks 557:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 2 5 theme well-functionalized 374:392 arg1 blocks 418:423 well-functionalized monosaccharide building blocks 374:423 well-functionalized monosaccharide building blocks 374:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 7 6 theme efficient 1158:1166 arg1 glycosylations 1168:1181 highly efficient glycosylations 1151:1181 highly efficient glycosylations 1151:1181 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26650028 1 7 theme optimized 165:173 arg1 strategy 193:200 an optimized modular synthetic strategy 162:200 an optimized modular synthetic strategy 162:200 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 7 8 used used 1132:1135 arg2 activators 1081:1090 cheaper activators 1073:1090 cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) 1073:1125 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26650028 7 9 theme assembly 1053:1060 arg1 reactions 1062:1070 the monosaccharide-to-oligosaccharide assembly reactions 1015:1070 the monosaccharide-to-oligosaccharide assembly reactions 1015:1070 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26650028 3 10 theme monosaccharide 561:574 arg1 blocks 585:590 the monosaccharide building blocks 557:590 the monosaccharide building blocks 557:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 3 11 theme glycosylation 538:550 arg1 step 530:533 every step 524:533 every step of glycosylation from the monosaccharide building blocks 524:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 3 12 from stereoselectivity 503:519 arg1 step 530:533 every step 524:533 every step of glycosylation from the monosaccharide building blocks 524:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 1 13 theme practical 64:72 arg1 synthesis 81:89 The practical formal synthesis 60:89 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 60:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 4 14 theme Efficient 593:601 arg1 routes 603:608 Efficient routes 593:608 Efficient routes to the syntheses of these fully functionalized building blocks 593:671 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 4 15 theme stage 716:720 arg1 modifications 739:751 oligosaccharide stage functional-group modifications 700:751 oligosaccharide stage functional-group modifications 700:751 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 1 16 theme fondaparinux 117:128 arg1 sodium 130:135 the anticoagulant drug fondaparinux sodium 1 94:137 the anticoagulant drug fondaparinux sodium 1 94:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 8 17 theme oligosaccharide 1243:1257 arg1 intermediates 1259:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 1 18 theme formal 74:79 arg1 synthesis 81:89 The practical formal synthesis 60:89 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 60:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 1 19 theme modular 175:181 arg1 strategy 193:200 an optimized modular synthetic strategy 162:200 an optimized modular synthetic strategy 162:200 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 5 20 theme blocks 784:789 arg1 syntheses 758:766 The syntheses 754:766 The syntheses of all building blocks 754:789 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 6 21 theme reactions 921:929 arg1 intermediates 883:895 common intermediates 876:895 common intermediates 876:895 In addition, common intermediates and a series of one-pot reactions were employed to enhance synthetic efficiency, improving the yield considerably. 26650028 6 21 theme reactions 921:929 arg1 series 903:908 a series 901:908 a series of one-pot reactions 901:929 In addition, common intermediates and a series of one-pot reactions were employed to enhance synthetic efficiency, improving the yield considerably. 26650028 2 22 theme sodium 282:287 arg1 synthesis 256:264 the synthesis 252:264 the synthesis of fondaparinux sodium 252:287 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 1 23 theme synthetic 183:191 arg1 strategy 193:200 an optimized modular synthetic strategy 162:200 an optimized modular synthetic strategy 162:200 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 5 24 theme expensive 843:851 arg1 reagents 853:860 expensive reagents 843:860 expensive reagents 843:860 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 6 25 theme one-pot 913:919 arg1 reactions 921:929 one-pot reactions 913:929 one-pot reactions 913:929 In addition, common intermediates and a series of one-pot reactions were employed to enhance synthetic efficiency, improving the yield considerably. 26650028 2 26 theme g 314:314 arg1 scale 316:320 a 10 g scale 309:320 a 10 g scale 309:320 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 4 27 theme building 657:664 arg1 blocks 666:671 these fully functionalized building blocks 630:671 these fully functionalized building blocks 630:671 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 0 28 theme Formal 0:5 arg1 Synthesis 7:15 Formal Synthesis 0:15 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium 0:57 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium. 26650028 3 29 theme building 576:583 arg1 blocks 585:590 the monosaccharide building blocks 557:590 the monosaccharide building blocks 557:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 3 30 from blocks 585:590 arg1 glycosylation 538:550 glycosylation 538:550 glycosylation from the monosaccharide building blocks 538:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 3 30 from blocks 585:590 arg1 step 530:533 every step 524:533 every step of glycosylation from the monosaccharide building blocks 524:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 2 31 theme overall 355:361 arg1 yield 363:367 3.5% overall yield 350:367 3.5% overall yield from well-functionalized monosaccharide building blocks 350:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 0 32 theme Anticoagulant 20:32 arg1 Sodium 52:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium. 26650028 5 33 theme reagents 853:860 arg1 use 836:838 the use 832:838 the use of expensive reagents 832:860 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 5 33 theme reagents 853:860 arg1 conditions 817:826 rigorous reaction conditions 799:826 rigorous reaction conditions 799:826 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 1 34 theme anticoagulant 98:110 arg1 sodium 130:135 the anticoagulant drug fondaparinux sodium 1 94:137 the anticoagulant drug fondaparinux sodium 1 94:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 8 35 theme scalable 1364:1371 arg1 synthesis 1373:1381 the scalable synthesis 1360:1381 the scalable synthesis of fondaparinux sodium 1360:1404 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 8 36 theme several 1216:1222 arg1 intermediates 1259:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 3 37 dep approach 478:485 arg1 2 466:466 2 466:466 2 466:466 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 1 38 theme drug 112:115 arg1 sodium 130:135 the anticoagulant drug fondaparinux sodium 1 94:137 the anticoagulant drug fondaparinux sodium 1 94:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 4 39 theme oligosaccharide 700:714 arg1 modifications 739:751 oligosaccharide stage functional-group modifications 700:751 oligosaccharide stage functional-group modifications 700:751 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 0 40 theme Fondaparinux 39:50 arg1 Sodium 52:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium. 26650028 8 41 theme fondaparinux 1386:1397 arg1 sodium 1399:1404 fondaparinux sodium 1386:1404 fondaparinux sodium 1386:1404 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 3 42 theme convergent 450:459 arg1 [3 461:462 a convergent [3 448:462 a convergent [3 448:462 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 2 43 theme building 409:416 arg1 blocks 418:423 well-functionalized monosaccharide building blocks 374:423 well-functionalized monosaccharide building blocks 374:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 0 44 theme Drug 34:37 arg1 Sodium 52:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Anticoagulant Drug Fondaparinux Sodium 20:57 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium. 26650028 8 45 theme purification 1298:1309 arg1 procedures 1311:1320 purification procedures 1298:1320 purification procedures 1298:1320 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 2 46 theme monosaccharide 394:407 arg1 blocks 418:423 well-functionalized monosaccharide building blocks 374:423 well-functionalized monosaccharide building blocks 374:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 8 47 theme sodium 1399:1404 arg1 synthesis 1373:1381 the scalable synthesis 1360:1381 the scalable synthesis of fondaparinux sodium 1360:1404 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 8 48 theme monosaccharide 1224:1237 arg1 intermediates 1259:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 several monosaccharide and oligosaccharide intermediates 1216:1271 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 7 49 theme cheaper 1073:1079 arg1 activators 1081:1090 cheaper activators 1073:1090 cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) 1073:1125 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26650028 2 50 theme collective 328:337 arg1 steps 339:343 14 collective steps 325:343 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks 325:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 0 51 theme Sodium 52:57 arg1 Synthesis 7:15 Formal Synthesis 0:15 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium 0:57 Formal Synthesis of Anticoagulant Drug Fondaparinux Sodium. 26650028 6 52 theme common 876:881 arg1 intermediates 883:895 common intermediates 876:895 common intermediates 876:895 In addition, common intermediates and a series of one-pot reactions were employed to enhance synthetic efficiency, improving the yield considerably. 26650028 3 53 theme excellent 493:501 arg1 stereoselectivity 503:519 excellent stereoselectivity 493:519 excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks 493:590 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 1 54 theme sodium 130:135 arg1 synthesis 81:89 The practical formal synthesis 60:89 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 60:137 The practical formal synthesis of the anticoagulant drug fondaparinux sodium 1 was accomplished using an optimized modular synthetic strategy. 26650028 2 55 with steps 339:343 arg1 yield 363:367 3.5% overall yield 350:367 3.5% overall yield from well-functionalized monosaccharide building blocks 350:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 2 56 theme fondaparinux 269:280 arg1 sodium 282:287 fondaparinux sodium 269:287 fondaparinux sodium 269:287 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 5 57 theme building 775:782 arg1 blocks 784:789 all building blocks 771:789 all building blocks 771:789 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 2 58 theme important 207:215 arg1 precursor 238:246 a precursor 236:246 a precursor for the synthesis of fondaparinux sodium 236:287 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 2 58 theme important 207:215 arg1 pentasaccharide 217:231 The important pentasaccharide 2 203:233 The important pentasaccharide 2 203:233 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 8 59 theme intermediates 1259:1271 arg1 crystallization 1197:1211 crystallization 1197:1211 crystallization of several monosaccharide and oligosaccharide intermediates 1197:1271 Furthermore, crystallization of several monosaccharide and oligosaccharide intermediates significantly simplified purification procedures, which would be greatly beneficial to the scalable synthesis of fondaparinux sodium. 26650028 3 60 theme coupling 469:476 arg1 approach 478:485 2] coupling approach 466:485 2] coupling approach 466:485 The strategy involved a convergent [3 + 2] coupling approach, with excellent stereoselectivity in every step of glycosylation from the monosaccharide building blocks. 26650028 4 61 theme functionalized 642:655 arg1 blocks 666:671 these fully functionalized building blocks 630:671 these fully functionalized building blocks 630:671 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 2 62 theme % 353:353 arg1 yield 363:367 3.5% overall yield 350:367 3.5% overall yield from well-functionalized monosaccharide building blocks 350:423 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 4 63 theme blocks 666:671 arg1 syntheses 617:625 the syntheses 613:625 the syntheses of these fully functionalized building blocks 613:671 Efficient routes to the syntheses of these fully functionalized building blocks were developed, minimizing oligosaccharide stage functional-group modifications. 26650028 5 64 theme rigorous 799:806 arg1 conditions 817:826 rigorous reaction conditions 799:826 rigorous reaction conditions 799:826 The syntheses of all building blocks avoided rigorous reaction conditions and the use of expensive reagents. 26650028 7 65 theme monosaccharide-to-oligosaccharide 1019:1051 arg1 reactions 1062:1070 the monosaccharide-to-oligosaccharide assembly reactions 1015:1070 the monosaccharide-to-oligosaccharide assembly reactions 1015:1070 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26650028 2 66 theme 3.5 350:352 arg1 % 353:353 % 353:353 % 353:353 The important pentasaccharide 2, a precursor for the synthesis of fondaparinux sodium, was synthesized on a 10 g scale in 14 collective steps with 3.5% overall yield from well-functionalized monosaccharide building blocks. 26650028 7 67 dep NIS/TfOH 1099:1106 arg1 e.g. 1093:1096 e.g. 1093:1096 e.g. 1093:1096 In the monosaccharide-to-oligosaccharide assembly reactions, cheaper activators (e.g., NIS/TfOH, TESOTf, and TfOH) were used to facilitate highly efficient glycosylations. 26286102 6 0 theme childhood 1067:1075 arg1 CDD 1102:1104 CDD 1102:1104 CDD 1102:1104 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 6 0 theme childhood 1067:1075 arg1 disorder 1092:1099 childhood disintegrative disorder 1067:1099 childhood disintegrative disorder (CDD) 1067:1105 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 10 1 theme onset 1536:1540 arg1 disturbance 1554:1564 the early onset psychiatric disturbance 1526:1564 the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS 1526:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 2 2 from deficiency 581:590 arg1 CNS 599:601 the CNS 595:601 the CNS 595:601 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 7 3 theme underlying 1194:1203 arg1 disorder 1223:1230 an underlying neurodegenerative disorder 1191:1230 an underlying neurodegenerative disorder 1191:1230 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 4 4 theme psychiatric 836:846 arg1 disturbances 848:859 psychiatric disturbances 836:859 psychiatric disturbances 836:859 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 3 5 theme HEXA 692:695 arg1 gene 697:700 the HEXA gene 688:700 the HEXA gene resulting in GM2-ganglioside storage in the CNS 688:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 10 6 theme protein 1609:1615 arg1 sialylation 1617:1627 defective protein sialylation 1599:1627 defective protein sialylation in the CNS 1599:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 11 7 theme neuropsychiatric 1829:1844 arg1 disorders 1846:1854 neuropsychiatric disorders 1829:1854 neuropsychiatric disorders 1829:1854 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 9 8 theme plasma 1440:1445 arg1 N-glycans 1447:1455 total plasma N-glycans 1434:1455 total plasma N-glycans 1434:1455 No changes of total plasma N-glycans were found. 26286102 8 9 theme triantennary 1333:1344 arg1 structures 1346:1355 sialylated biantennary and triantennary structures 1306:1355 sialylated biantennary and triantennary structures 1306:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 3 10 theme GM2-ganglioside 715:729 arg1 storage 731:737 GM2-ganglioside storage 715:737 GM2-ganglioside storage in the CNS 715:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 1 11 theme moiety 226:231 arg1 processing 192:201 processing 192:201 processing 192:201 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 11 theme moiety 226:231 arg1 synthesis 178:186 synthesis 178:186 synthesis 178:186 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 3 12 from mutations 675:683 arg1 gene 697:700 the HEXA gene 688:700 the HEXA gene resulting in GM2-ganglioside storage in the CNS 688:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 11 13 theme protein 1869:1875 arg1 glycosylation 1877:1889 unproper protein glycosylation 1860:1889 unproper protein glycosylation in the CNS 1860:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 2 14 theme deficiency 581:590 arg1 presence 557:564 the presence 553:564 the presence of sialylation deficiency in the CNS 553:601 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 8 15 theme CSF 1262:1264 arg1 N-glycans 1266:1274 CSF N-glycans 1262:1274 CSF N-glycans 1262:1274 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 3 16 theme Tay 615:617 arg1 disorder 656:663 a lysosomal disorder 644:663 a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS 644:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 16 theme Tay 615:617 arg1 LOTSD 634:638 LOTSD 634:638 LOTSD 634:638 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 16 theme Tay 615:617 arg1 disease 625:631 Late-onset Tay Sachs disease 604:631 Late-onset Tay Sachs disease (LOTSD) 604:639 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 0 17 theme GM2 70:72 arg1 gangliosidosis 74:87 GM2 gangliosidosis 70:87 GM2 gangliosidosis 70:87 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 5 18 theme cerebrospinal 902:914 arg1 CSF 923:925 CSF 923:925 CSF 923:925 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 5 18 theme cerebrospinal 902:914 arg1 fluid 916:920 cerebrospinal fluid 902:920 the cerebrospinal fluid (CSF) 898:926 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 8 19 theme sialylated 1306:1315 arg1 structures 1346:1355 sialylated biantennary and triantennary structures 1306:1355 sialylated biantennary and triantennary structures 1306:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 11 20 theme GM2-gangliosidosis 1769:1786 arg1 pathomechanisms 1750:1764 possible pathomechanisms 1741:1764 possible pathomechanisms of GM2-gangliosidosis 1741:1786 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 2 21 theme spectrum 525:532 arg1 ASD 545:547 ASD 545:547 ASD 545:547 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 2 21 theme spectrum 525:532 arg1 disorders 534:542 autism spectrum disorders 518:542 autism spectrum disorders (ASD) 518:548 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 0 22 theme childhood 103:111 arg1 disorder 128:135 childhood disintegrative disorder 103:135 childhood disintegrative disorder 103:135 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 2 23 theme sialylation 458:468 arg1 occurrence 504:513 the occurrence 500:513 the occurrence of autism spectrum disorders (ASD) 500:548 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 2 23 theme sialylation 458:468 arg1 importance 424:433 the importance 420:433 the importance of proper glycoprotein sialylation for the synaptic function 420:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 10 24 gly sialylation 1617:1627 arg1 CNS 1636:1638 the CNS 1632:1638 the CNS 1632:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 10 24 gly sialylation 1617:1627 arg1 patient 1587:1593 the patient 1583:1593 the patient 1583:1593 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 8 25 theme structures 1346:1355 arg1 increase 1364:1371 an increase 1361:1371 an increase of asialo-core fucosylated bisected N-glycans 1361:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 8 25 theme structures 1346:1355 arg1 decrease 1294:1301 a dramatic decrease 1283:1301 a dramatic decrease of sialylated biantennary and triantennary structures 1283:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 0 26 theme CSF 0:2 arg1 profile 13:19 CSF N-glycan profile 0:19 CSF N-glycan profile 0:19 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 1 27 theme several 278:284 arg1 functions 286:294 several functions 278:294 several functions for the proper central nervous system (CNS) development and function 278:363 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 6 28 theme regressive 1025:1034 arg1 autism 1036:1041 regressive autism 1025:1041 regressive autism fulfilling criteria for childhood disintegrative disorder (CDD) 1025:1105 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 2 29 theme proper 438:443 arg1 sialylation 458:468 proper glycoprotein sialylation 438:468 proper glycoprotein sialylation for the synaptic function 438:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 10 30 theme possible 1495:1502 arg1 relationships 1504:1516 possible relationships 1495:1516 possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS 1495:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 5 31 theme year-old 936:943 arg1 patient 945:951 a 14 year-old patient 931:951 a 14 year-old patient with GM2-gangliosidosis (LOTSD) 931:983 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 2 32 from CNS 599:601 arg1 presence 557:564 the presence 553:564 the presence of sialylation deficiency in the CNS 553:601 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 11 33 theme CSF 1710:1712 arg1 proteins 1714:1721 CSF proteins 1710:1721 CSF proteins 1710:1721 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 11 34 gly glycosylation 1877:1889 arg1 CNS 1898:1900 the CNS 1894:1900 the CNS 1894:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 0 35 theme sialylation 29:39 arg1 deficiency 41:50 sialylation deficiency 29:50 sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder 29:135 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 1 36 theme proper 304:309 arg1 development 340:350 the proper central nervous system (CNS) development 300:350 the proper central nervous system (CNS) development 300:350 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 10 37 theme early 1530:1534 arg1 disturbance 1554:1564 the early onset psychiatric disturbance 1526:1564 the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS 1526:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 8 38 theme fucosylated 1388:1398 arg1 N-glycans 1409:1417 asialo-core fucosylated bisected N-glycans 1376:1417 asialo-core fucosylated bisected N-glycans 1376:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 7 39 theme CSF 1110:1112 arg1 sample 1114:1119 A CSF sample 1108:1119 A CSF sample 1108:1119 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 1 40 theme nervous 319:325 arg1 CNS 335:337 CNS 335:337 CNS 335:337 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 40 theme nervous 319:325 arg1 system 327:332 central nervous system 311:332 the proper central nervous system (CNS) development 300:350 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 8 41 theme N-glycans 1409:1417 arg1 increase 1364:1371 an increase 1361:1371 an increase of asialo-core fucosylated bisected N-glycans 1361:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 8 41 theme N-glycans 1409:1417 arg1 decrease 1294:1301 a dramatic decrease 1283:1301 a dramatic decrease of sialylated biantennary and triantennary structures 1283:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 3 42 theme lysosomal 646:654 arg1 disorder 656:663 a lysosomal disorder 644:663 a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS 644:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 42 theme lysosomal 646:654 arg1 disease 625:631 Late-onset Tay Sachs disease 604:631 Late-onset Tay Sachs disease (LOTSD) 604:639 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 4 43 theme neurological 786:797 arg1 impairment 799:808 progressive neurological impairment 774:808 progressive neurological impairment 774:808 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 2 44 theme Previous 366:373 arg1 studies 401:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 6 45 theme disintegrative 1077:1090 arg1 CDD 1102:1104 CDD 1102:1104 CDD 1102:1104 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 6 45 theme disintegrative 1077:1090 arg1 disorder 1092:1099 childhood disintegrative disorder 1067:1099 childhood disintegrative disorder (CDD) 1067:1105 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 10 46 theme psychiatric 1542:1552 arg1 disturbance 1554:1564 the early onset psychiatric disturbance 1526:1564 the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS 1526:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 6 47 attach presented 1015:1023 arg1 age 993:995 the age 989:995 the age of 4 989:1000 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 6 47 attach presented 1015:1023 arg2 patient 1007:1013 the patient 1003:1013 the patient 1003:1013 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 10 48 from sialylation 1617:1627 arg1 CNS 1636:1638 the CNS 1632:1638 the CNS 1632:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 10 48 from sialylation 1617:1627 arg1 patient 1587:1593 the patient 1583:1593 the patient 1583:1593 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 11 49 from disorders 1846:1854 arg1 CNS 1898:1900 the CNS 1894:1900 the CNS 1894:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 7 50 theme neurodegenerative 1205:1221 arg1 disorder 1223:1230 an underlying neurodegenerative disorder 1191:1230 an underlying neurodegenerative disorder 1191:1230 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 2 51 from presence 557:564 arg1 CNS 599:601 the CNS 595:601 the CNS 595:601 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 11 52 from glycosylation 1877:1889 arg1 CNS 1898:1900 the CNS 1894:1900 the CNS 1894:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 11 53 theme aberrant 1678:1685 arg1 structures 1696:1705 aberrant N-glycan structures 1678:1705 aberrant N-glycan structures of CSF proteins in LOTSD 1678:1730 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 0 54 with patient 57:63 arg1 gangliosidosis 74:87 GM2 gangliosidosis 70:87 GM2 gangliosidosis 70:87 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 9 55 theme total 1434:1438 arg1 N-glycans 1447:1455 total plasma N-glycans 1434:1455 total plasma N-glycans 1434:1455 No changes of total plasma N-glycans were found. 26286102 4 56 theme disturbances 848:859 arg1 impairment 799:808 progressive neurological impairment 774:808 progressive neurological impairment 774:808 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 4 56 theme disturbances 848:859 arg1 co-occurrence 819:831 high co-occurrence 814:831 high co-occurrence 814:831 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 10 57 theme defective 1599:1607 arg1 sialylation 1617:1627 defective protein sialylation 1599:1627 defective protein sialylation in the CNS 1599:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 4 58 theme high 814:817 arg1 co-occurrence 819:831 high co-occurrence 814:831 high co-occurrence 814:831 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 1 59 theme oligosaccharide 210:224 arg1 moiety 226:231 the oligosaccharide moiety 206:231 the oligosaccharide moiety (N-glycan) linked to a protein 206:262 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 59 theme oligosaccharide 210:224 arg1 N-glycan 234:241 N-glycan 234:241 N-glycan 234:241 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 60 attach linked 244:249 arg2 N-glycan 234:241 N-glycan 234:241 N-glycan 234:241 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 60 attach linked 244:249 arg2 moiety 226:231 the oligosaccharide moiety 206:231 the oligosaccharide moiety (N-glycan) linked to a protein 206:262 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 60 attach linked 244:249 arg1 protein 256:262 a protein 254:262 a protein 254:262 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 5 61 theme N-glycome 877:885 arg1 profile 887:893 the N-glycome profile 873:893 the N-glycome profile of the cerebrospinal fluid (CSF) 873:926 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 8 62 theme definite 1242:1249 arg1 changes 1251:1257 definite changes 1242:1257 definite changes of CSF N-glycans 1242:1274 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 5 63 with patient 945:951 arg1 LOTSD 978:982 LOTSD 978:982 LOTSD 978:982 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 5 63 with patient 945:951 arg1 GM2-gangliosidosis 958:975 GM2-gangliosidosis 958:975 GM2-gangliosidosis (LOTSD) 958:983 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 5 64 theme fluid 916:920 arg1 profile 887:893 the N-glycome profile 873:893 the N-glycome profile of the cerebrospinal fluid (CSF) 873:926 We studied the N-glycome profile of the cerebrospinal fluid (CSF) in a 14 year-old patient with GM2-gangliosidosis (LOTSD). 26286102 2 65 theme sialylation 569:579 arg1 deficiency 581:590 sialylation deficiency 569:590 sialylation deficiency in the CNS 569:601 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 11 66 theme unproper 1860:1867 arg1 glycosylation 1877:1889 unproper protein glycosylation 1860:1889 unproper protein glycosylation in the CNS 1860:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 3 67 theme Sachs 619:623 arg1 disorder 656:663 a lysosomal disorder 644:663 a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS 644:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 67 theme Sachs 619:623 arg1 LOTSD 634:638 LOTSD 634:638 LOTSD 634:638 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 67 theme Sachs 619:623 arg1 disease 625:631 Late-onset Tay Sachs disease 604:631 Late-onset Tay Sachs disease (LOTSD) 604:639 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 1 68 theme Protein 138:144 arg1 N-glycosylation 146:160 Protein N-glycosylation 138:160 Protein N-glycosylation 138:160 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 6 69 theme 4 1000:1000 arg1 age 993:995 the age 989:995 the age of 4 989:1000 At the age of 4, the patient presented regressive autism fulfilling criteria for childhood disintegrative disorder (CDD). 26286102 8 70 theme N-glycans 1266:1274 arg1 changes 1251:1257 definite changes 1242:1257 definite changes of CSF N-glycans 1242:1274 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 3 71 theme Late-onset 604:613 arg1 disorder 656:663 a lysosomal disorder 644:663 a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS 644:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 71 theme Late-onset 604:613 arg1 LOTSD 634:638 LOTSD 634:638 LOTSD 634:638 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 3 71 theme Late-onset 604:613 arg1 disease 625:631 Late-onset Tay Sachs disease 604:631 Late-onset Tay Sachs disease (LOTSD) 604:639 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 9 72 theme N-glycans 1447:1455 arg1 changes 1423:1429 No changes 1420:1429 No changes of total plasma N-glycans 1420:1455 No changes of total plasma N-glycans were found. 26286102 11 73 from structures 1696:1705 arg1 LOTSD 1726:1730 LOTSD 1726:1730 LOTSD 1726:1730 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 11 74 theme possible 1741:1748 arg1 pathomechanisms 1750:1764 possible pathomechanisms 1741:1764 possible pathomechanisms of GM2-gangliosidosis 1741:1786 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 8 75 theme dramatic 1285:1292 arg1 decrease 1294:1301 a dramatic decrease 1283:1301 a dramatic decrease of sialylated biantennary and triantennary structures 1283:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 11 76 theme existing 1798:1805 arg1 relationships 1807:1819 existing relationships 1798:1819 existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS 1798:1900 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 7 77 theme work-up 1162:1168 arg1 course 1141:1146 the course 1137:1146 the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder 1137:1230 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 2 78 theme disorders 534:542 arg1 occurrence 504:513 the occurrence 500:513 the occurrence of autism spectrum disorders (ASD) 500:548 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 2 78 theme disorders 534:542 arg1 importance 424:433 the importance 420:433 the importance of proper glycoprotein sialylation for the synaptic function 420:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 0 79 theme disintegrative 113:126 arg1 disorder 128:135 childhood disintegrative disorder 103:135 childhood disintegrative disorder 103:135 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 2 80 theme autism 518:523 arg1 ASD 545:547 ASD 545:547 ASD 545:547 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 2 80 theme autism 518:523 arg1 disorders 534:542 autism spectrum disorders 518:542 autism spectrum disorders (ASD) 518:548 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 0 81 from deficiency 41:50 arg1 patient 57:63 a patient 55:63 a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder 55:135 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 3 82 from storage 731:737 arg1 CNS 746:748 the CNS 742:748 the CNS 742:748 Late-onset Tay Sachs disease (LOTSD) is a lysosomal disorder caused by mutations in the HEXA gene resulting in GM2-ganglioside storage in the CNS. 26286102 8 83 theme biantennary 1317:1327 arg1 structures 1346:1355 sialylated biantennary and triantennary structures 1306:1355 sialylated biantennary and triantennary structures 1306:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 2 84 theme clinical 392:399 arg1 studies 401:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 0 85 theme N-glycan 4:11 arg1 profile 13:19 CSF N-glycan profile 0:19 CSF N-glycan profile 0:19 CSF N-glycan profile reveals sialylation deficiency in a patient with GM2 gangliosidosis presenting as childhood disintegrative disorder. 26286102 2 86 theme glycoprotein 445:456 arg1 sialylation 458:468 proper glycoprotein sialylation 438:468 proper glycoprotein sialylation for the synaptic function 438:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 1 87 dep synthesis 178:186 arg1 the 174:176 the 174:176 the 174:176 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 2 88 theme experimental 375:386 arg1 studies 401:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies 366:407 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 8 89 gly sialylated 1306:1315 arg1 structures 1346:1355 sialylated biantennary and triantennary structures 1306:1355 sialylated biantennary and triantennary structures 1306:1355 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 2 90 theme synaptic 478:485 arg1 function 487:494 the synaptic function 474:494 the synaptic function 474:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 11 91 theme N-glycan 1687:1694 arg1 structures 1696:1705 aberrant N-glycan structures 1678:1705 aberrant N-glycan structures of CSF proteins in LOTSD 1678:1730 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 7 92 theme diagnostic 1151:1160 arg1 work-up 1162:1168 diagnostic work-up 1151:1168 diagnostic work-up for the suspicion of an underlying neurodegenerative disorder 1151:1230 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 1 93 theme central 311:317 arg1 CNS 335:337 CNS 335:337 CNS 335:337 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 1 93 theme central 311:317 arg1 system 327:332 central nervous system 311:332 the proper central nervous system (CNS) development 300:350 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 8 94 theme asialo-core 1376:1386 arg1 N-glycans 1409:1417 asialo-core fucosylated bisected N-glycans 1376:1417 asialo-core fucosylated bisected N-glycans 1376:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 1 95 theme system 327:332 arg1 development 340:350 the proper central nervous system (CNS) development 300:350 the proper central nervous system (CNS) development 300:350 Protein N-glycosylation consists in the synthesis and processing of the oligosaccharide moiety (N-glycan) linked to a protein and it serves several functions for the proper central nervous system (CNS) development and function. 26286102 10 96 from CDD 1576:1578 arg1 CNS 1636:1638 the CNS 1632:1638 the CNS 1632:1638 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 10 96 from CDD 1576:1578 arg1 patient 1587:1593 the patient 1583:1593 the patient 1583:1593 Herein findings highlight possible relationships between the early onset psychiatric disturbance featuring CDD in the patient and defective protein sialylation in the CNS. 26286102 2 97 gly glycoprotein 445:456 arg1 glycoprotein 445:456 proper glycoprotein sialylation 438:468 proper glycoprotein sialylation for the synaptic function 438:494 Previous experimental and clinical studies have shown the importance of proper glycoprotein sialylation for the synaptic function and the occurrence of autism spectrum disorders (ASD) in the presence of sialylation deficiency in the CNS. 26286102 11 98 theme proteins 1714:1721 arg1 structures 1696:1705 aberrant N-glycan structures 1678:1705 aberrant N-glycan structures of CSF proteins in LOTSD 1678:1730 In conclusion, the study first shows aberrant N-glycan structures of CSF proteins in LOTSD; unveils possible pathomechanisms of GM2-gangliosidosis; supports existing relationships between neuropsychiatric disorders and unproper protein glycosylation in the CNS. 26286102 8 99 theme bisected 1400:1407 arg1 N-glycans 1409:1417 asialo-core fucosylated bisected N-glycans 1376:1417 asialo-core fucosylated bisected N-glycans 1376:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26286102 7 100 theme disorder 1223:1230 arg1 suspicion 1178:1186 the suspicion 1174:1186 the suspicion of an underlying neurodegenerative disorder 1174:1230 A CSF sample was obtained in the course of diagnostic work-up for the suspicion of an underlying neurodegenerative disorder. 26286102 4 101 theme progressive 774:784 arg1 impairment 799:808 progressive neurological impairment 774:808 progressive neurological impairment 774:808 It is characterized by progressive neurological impairment and high co-occurrence of psychiatric disturbances. 26286102 8 102 gly fucosylated 1388:1398 arg1 N-glycans 1409:1417 asialo-core fucosylated bisected N-glycans 1376:1417 asialo-core fucosylated bisected N-glycans 1376:1417 We found definite changes of CSF N-glycans due to a dramatic decrease of sialylated biantennary and triantennary structures and an increase of asialo-core fucosylated bisected N-glycans. 26082233 5 0 theme cell 963:966 arg1 surface 968:974 the cell surface 959:974 the cell surface 959:974 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 4 1 theme new 636:638 arg1 analogues 655:663 therefore new monosaccharide analogues 626:663 therefore new monosaccharide analogues 626:663 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 0 2 from Glyco-Engineering 10:26 arg1 Cells 42:46 Eukaryotic Cells 31:46 Eukaryotic Cells 31:46 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications. 26082233 0 2 from Glyco-Engineering 10:26 arg1 Applications 61:72 Selected Applications 52:72 Selected Applications 52:72 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications. 26082233 8 3 theme employed 1401:1408 arg1 introduction 1460:1471 the introduction 1456:1471 the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells 1456:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 3 theme employed 1401:1408 arg1 application 1410:1420 The most useful and most frequently employed application 1365:1420 The most useful and most frequently employed application of metabolic glyco-engineering 1365:1451 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 7 4 theme structures 1282:1291 arg1 synthesis 1262:1270 the synthesis 1258:1270 the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling 1258:1362 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 8 5 theme cells 1602:1606 arg1 surface 1586:1592 the surface 1582:1592 the surface of live cells 1582:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 5 6 from surface 968:974 arg1 glycans 948:954 some glycans 943:954 some glycans at the cell surface 943:974 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 8 7 theme metabolic 1425:1433 arg1 glyco-engineering 1435:1451 metabolic glyco-engineering 1425:1451 metabolic glyco-engineering 1425:1451 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 4 8 theme monosaccharide 640:653 arg1 analogues 655:663 therefore new monosaccharide analogues 626:663 therefore new monosaccharide analogues 626:663 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 7 9 theme cell 1332:1335 arg1 surface 1337:1343 the cell surface 1328:1343 the cell surface 1328:1343 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 5 10 theme monosaccharide 901:914 arg1 analogues 916:924 the monosaccharide analogues 897:924 the monosaccharide analogues 897:924 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 1 11 theme metabolic 78:86 arg1 glycoconjugates 115:129 metabolic glyco-engineering cellular glycoconjugates 78:129 metabolic glyco-engineering cellular glycoconjugates 78:129 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 6 12 theme specific 1140:1147 arg1 sugars 1149:1154 specific sugars 1140:1154 specific sugars 1140:1154 The presence of those glycans can be easily and quantitatively detected by lectin binding or by chemical methods identifying specific sugars. 26082233 6 13 theme chemical 1111:1118 arg1 methods 1120:1126 chemical methods 1111:1126 chemical methods identifying specific sugars 1111:1154 The presence of those glycans can be easily and quantitatively detected by lectin binding or by chemical methods identifying specific sugars. 26082233 7 14 theme sugar 1221:1225 arg1 incorporation 1227:1239 sugar incorporation 1221:1239 sugar incorporation 1221:1239 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 1 15 theme glyco-engineering 88:104 arg1 glycoconjugates 115:129 metabolic glyco-engineering cellular glycoconjugates 78:129 metabolic glyco-engineering cellular glycoconjugates 78:129 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 4 16 from suitability 711:721 arg1 glyco-engineering 736:752 metabolic glyco-engineering 726:752 metabolic glyco-engineering 726:752 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 7 17 theme Monosaccharide 1157:1170 arg1 analogues 1172:1180 Monosaccharide analogues 1157:1180 Monosaccharide analogues 1157:1180 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 1 18 theme cellular 106:113 arg1 glycoconjugates 115:129 metabolic glyco-engineering cellular glycoconjugates 78:129 metabolic glyco-engineering cellular glycoconjugates 78:129 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 4 19 theme metabolic 726:734 arg1 glyco-engineering 736:752 metabolic glyco-engineering 726:752 metabolic glyco-engineering 726:752 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 0 20 theme Metabolic 0:8 arg1 Glyco-Engineering 10:26 Metabolic Glyco-Engineering 0:26 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications 0:72 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications. 26082233 2 21 theme activated 411:419 arg1 sugars 421:426 activated sugars 411:426 activated sugars 411:426 In order to get incorporated, the monosaccharides need to enter the cytoplasm and to be substrates for the enzymes necessary for their transformation into activated sugars, most often nucleotide sugars. 26082233 0 22 theme Eukaryotic 31:40 arg1 Cells 42:46 Eukaryotic Cells 31:46 Eukaryotic Cells 31:46 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications. 26082233 6 23 theme glycans 1037:1043 arg1 presence 1019:1026 The presence 1015:1026 The presence of those glycans 1015:1043 The presence of those glycans can be easily and quantitatively detected by lectin binding or by chemical methods identifying specific sugars. 26082233 8 24 theme useful 1374:1379 arg1 introduction 1460:1471 the introduction 1456:1471 the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells 1456:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 24 theme useful 1374:1379 arg1 application 1410:1420 The most useful and most frequently employed application 1365:1420 The most useful and most frequently employed application of metabolic glyco-engineering 1365:1451 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 7 25 from surface 1337:1343 arg1 detectable 1314:1323 detectable 1314:1323 detectable 1314:1323 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 8 26 theme reactive 1476:1483 arg1 groups 1485:1490 reactive groups 1476:1490 reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells 1476:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 4 27 theme tissue 686:691 arg1 culture 693:699 tissue culture 686:699 tissue culture 686:699 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 5 28 theme glycosylation 765:777 arg1 appropriate 800:810 appropriate 800:810 appropriate 800:810 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 5 28 theme glycosylation 765:777 arg1 mutants 779:785 glycosylation mutants 765:785 glycosylation mutants 765:785 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 8 29 theme efficient 1549:1557 arg1 labeling 1559:1566 the efficient labeling 1545:1566 the efficient labeling of glycans 1545:1577 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 1 30 theme sugars 248:253 arg1 monosaccharides 183:197 synthetic monosaccharides 173:197 synthetic monosaccharides which are usually analogues of naturally present sugars 173:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 1 30 theme sugars 248:253 arg1 analogues 217:225 analogues 217:225 analogues of naturally present sugars 217:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 5 31 theme analogues 916:924 arg1 introduction 881:892 the introduction 877:892 the introduction of the monosaccharide analogues 877:924 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 2 32 theme nucleotide 440:449 arg1 transformation 391:404 their transformation 385:404 their transformation into activated sugars 385:426 In order to get incorporated, the monosaccharides need to enter the cytoplasm and to be substrates for the enzymes necessary for their transformation into activated sugars, most often nucleotide sugars. 26082233 2 32 theme nucleotide 440:449 arg1 sugars 451:456 nucleotide sugars 440:456 most often nucleotide sugars 429:456 In order to get incorporated, the monosaccharides need to enter the cytoplasm and to be substrates for the enzymes necessary for their transformation into activated sugars, most often nucleotide sugars. 26082233 5 33 theme specific 848:855 arg1 glycans 857:863 specific glycans 848:863 specific glycans 848:863 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 8 34 theme groups 1485:1490 arg1 introduction 1460:1471 the introduction 1456:1471 the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells 1456:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 34 theme groups 1485:1490 arg1 application 1410:1420 The most useful and most frequently employed application 1365:1420 The most useful and most frequently employed application of metabolic glyco-engineering 1365:1451 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 4 35 theme Such 569:572 arg1 pathways 574:581 Such pathways 569:581 Such pathways 569:581 Such pathways are difficult to reconstitute in vitro and therefore new monosaccharide analogues have to be tested in tissue culture for their suitability in metabolic glyco-engineering. 26082233 7 36 theme glycan 1275:1280 arg1 structures 1282:1291 glycan structures 1275:1291 glycan structures which is also easily detectable at the cell surface by lectin labeling 1275:1362 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 2 37 theme necessary 371:379 arg1 enzymes 363:369 the enzymes 359:369 the enzymes necessary for their transformation into activated sugars, most often nucleotide sugars 359:456 In order to get incorporated, the monosaccharides need to enter the cytoplasm and to be substrates for the enzymes necessary for their transformation into activated sugars, most often nucleotide sugars. 26082233 0 38 theme Selected 52:59 arg1 Applications 61:72 Selected Applications 52:72 Selected Applications 52:72 Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications. 26082233 8 39 theme glycans 1571:1577 arg1 labeling 1559:1566 the efficient labeling 1545:1566 the efficient labeling of glycans 1545:1577 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 40 theme click 1525:1529 arg1 reactions 1531:1539 bio-orthogonal click reactions 1510:1539 bio-orthogonal click reactions for the efficient labeling of glycans 1510:1577 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 5 41 theme unnatural 985:993 arg1 sugar 995:999 the unnatural sugar 981:999 the unnatural sugar incorporated 981:1012 For this, glycosylation mutants are the most appropriate since they are unable to synthesize specific glycans but through the introduction of the monosaccharide analogues they may express some glycans at the cell surface with the unnatural sugar incorporated. 26082233 8 42 theme glyco-engineering 1435:1451 arg1 introduction 1460:1471 the introduction 1456:1471 the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells 1456:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 42 theme glyco-engineering 1435:1451 arg1 application 1410:1420 The most useful and most frequently employed application 1365:1420 The most useful and most frequently employed application of metabolic glyco-engineering 1365:1451 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 8 43 theme bio-orthogonal 1510:1523 arg1 reactions 1531:1539 bio-orthogonal click reactions 1510:1539 bio-orthogonal click reactions for the efficient labeling of glycans 1510:1577 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 7 44 theme lectin 1348:1353 arg1 labeling 1355:1362 lectin labeling 1348:1362 lectin labeling 1348:1362 Monosaccharide analogues can also block the pathways leading to sugar incorporation, thus inhibiting the synthesis of glycan structures which is also easily detectable at the cell surface by lectin labeling. 26082233 6 45 theme lectin 1090:1095 arg1 binding 1097:1103 lectin binding 1090:1103 lectin binding 1090:1103 The presence of those glycans can be easily and quantitatively detected by lectin binding or by chemical methods identifying specific sugars. 26082233 1 46 theme present 240:246 arg1 sugars 248:253 naturally present sugars 230:253 naturally present sugars 230:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 1 47 theme synthetic 173:181 arg1 monosaccharides 183:197 synthetic monosaccharides 173:197 synthetic monosaccharides which are usually analogues of naturally present sugars 173:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 1 47 theme synthetic 173:181 arg1 analogues 217:225 analogues 217:225 analogues of naturally present sugars 217:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 26082233 8 48 theme live 1597:1600 arg1 cells 1602:1606 live cells 1597:1606 live cells 1597:1606 The most useful and most frequently employed application of metabolic glyco-engineering is the introduction of reactive groups which can undergo bio-orthogonal click reactions for the efficient labeling of glycans at the surface of live cells. 26082233 1 49 theme monosaccharides 183:197 arg1 incorporation 156:168 the incorporation 152:168 the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars 152:253 By metabolic glyco-engineering cellular glycoconjugates are modified through the incorporation of synthetic monosaccharides which are usually analogues of naturally present sugars. 25552259 6 0 theme essential 777:785 arg1 role 787:790 an essential role 774:790 an essential role 774:790 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 2 1 theme glycoprotein 337:348 arg1 O- 285:286 O- 285:286 O- 285:286 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 1 theme glycoprotein 337:348 arg1 chains 305:310 N-glycosidic chains 292:310 N-glycosidic chains 292:310 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 1 2 theme blood 259:263 arg1 antigens 271:278 the Gerbich blood group antigens 247:278 the Gerbich blood group antigens 247:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 9 3 gly heterogeneity 1241:1253 arg1 N-glycans 1266:1274 the GPC N-glycans 1258:1274 the GPC N-glycans 1258:1274 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 11 4 theme EBA-140 1671:1677 arg1 ligand 1679:1684 the EBA-140 ligand 1667:1684 the EBA-140 ligand 1667:1684 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 10 5 theme data 1425:1428 arg1 interpretation 1403:1416 new interpretation 1399:1416 new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC 1399:1490 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 3 6 theme Amino-acid 488:497 arg1 sequence 499:506 Amino-acid sequence 488:506 Amino-acid sequence 488:506 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 6 theme Amino-acid 488:497 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 6 7 theme merozoite 846:854 arg1 ligand 856:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 9 8 theme glycophorin 1335:1345 arg1 N-glycans 1349:1357 glycophorin A N-glycans 1335:1357 glycophorin A N-glycans 1335:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 11 9 theme GPC 1805:1807 arg1 structure 1818:1826 the GPC N-glycan structure 1801:1826 the GPC N-glycan structure 1801:1826 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 2 10 theme N-glycosidic 292:303 arg1 chains 305:310 N-glycosidic chains 292:310 N-glycosidic chains 292:310 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 1 11 theme glycosylated 163:174 arg1 sialoglycoproteins 185:202 highly glycosylated integral sialoglycoproteins 156:202 highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens 156:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 11 theme glycosylated 163:174 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 10 12 theme new 1399:1401 arg1 interpretation 1403:1416 new interpretation 1399:1416 new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC 1399:1490 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 5 13 theme N-glycans 666:674 arg1 structure 649:657 the structure 645:657 the structure of GPC N-glycans 645:674 are well characterized but the structure of GPC N-glycans has remained unknown. 25552259 9 14 theme structural 1230:1239 arg1 heterogeneity 1241:1253 structural heterogeneity 1230:1253 structural heterogeneity of the GPC N-glycans 1230:1274 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 3 15 theme attachment 528:537 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 15 theme attachment 528:537 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 15 theme attachment 528:537 arg1 sites 539:543 oligosaccharide attachment sites 512:543 oligosaccharide attachment sites 512:543 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 9 16 located found 1326:1330 arg1 N-glycans 1349:1357 glycophorin A N-glycans 1335:1357 glycophorin A N-glycans 1335:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 9 16 located found 1326:1330 arg2 elements 1313:1320 structural elements 1302:1320 structural elements not found in glycophorin A N-glycans 1302:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 1 17 theme human 207:211 arg1 membranes 228:236 human red blood cell membranes 207:236 human red blood cell membranes carrying the Gerbich blood group antigens 207:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 9 18 theme GPC 1262:1264 arg1 N-glycans 1266:1274 the GPC N-glycans 1258:1274 the GPC N-glycans 1258:1274 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 3 19 theme erythrocyte 554:564 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 7 20 theme EBA-140 974:980 arg1 ligand 982:987 the EBA-140 ligand 970:987 the EBA-140 ligand 970:987 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 11 21 contain containing 1569:1578 arg2 units 1602:1606 repeating lactosamine units 1580:1606 repeating lactosamine units of the GPC Gerbich variant 1580:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 21 contain containing 1569:1578 arg1 chains 1562:1567 N-glycosidic chains 1549:1567 N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1549:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 0 22 theme putative 45:52 arg1 N-glycans 32:40 glycophorin C N-glycans 18:40 glycophorin C N-glycans 18:40 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 0 22 theme putative 45:52 arg1 component 54:62 a putative component 43:62 a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand 43:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 1 23 contain carrying 238:245 arg1 membranes 228:236 human red blood cell membranes 207:236 human red blood cell membranes carrying the Gerbich blood group antigens 207:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 23 contain carrying 238:245 arg2 antigens 271:278 the Gerbich blood group antigens 247:278 the Gerbich blood group antigens 247:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 7 24 theme binding 953:959 arg1 site 961:964 the GPC binding site 945:964 the GPC binding site for the EBA-140 ligand 945:987 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 11 25 theme terminal 1524:1531 arg1 fucosylation 1533:1544 preferable terminal fucosylation 1513:1544 preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1513:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 25 theme terminal 1524:1531 arg1 explanation 1647:1657 an explanation 1644:1657 an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure 1644:1826 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 6 26 theme GPC 755:757 arg1 N-glycans 759:767 GPC N-glycans 755:767 GPC N-glycans 755:767 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 0 27 theme GPC 71:73 arg1 site 84:87 the GPC receptor site 67:87 the GPC receptor site for Plasmodium falciparum EBA-140 ligand 67:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 9 28 theme elements 1313:1320 arg1 existence 1289:1297 the existence 1285:1297 the existence of structural elements not found in glycophorin A N-glycans 1285:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 8 29 theme detailed 1007:1014 arg1 analysis 1027:1034 detailed structural analysis 1007:1034 detailed structural analysis using sequential mass spectrometry 1007:1069 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 10 30 theme ligand 1478:1483 arg1 binding 1445:1451 the binding 1441:1451 the binding of P. falciparum EBA-140 ligand to GPC 1441:1490 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 0 31 theme site 84:87 arg1 N-glycans 32:40 glycophorin C N-glycans 18:40 glycophorin C N-glycans 18:40 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 0 31 theme site 84:87 arg1 component 54:62 a putative component 43:62 a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand 43:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 10 32 dep P. 1456:1457 arg1 falciparum 1459:1468 falciparum 1459:1468 falciparum 1459:1468 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 1 33 theme group 265:269 arg1 antigens 271:278 the Gerbich blood group antigens 247:278 the Gerbich blood group antigens 247:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 34 theme sequential 1042:1051 arg1 spectrometry 1058:1069 sequential mass spectrometry 1042:1069 sequential mass spectrometry 1042:1069 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 2 35 gly glycoprotein 337:348 arg1 properties 379:388 Antigenic properties 369:388 Antigenic properties of human glycophorins 369:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 35 gly glycoprotein 337:348 arg1 glycoprotein 337:348 the major erythrocyte glycoprotein 315:348 the major erythrocyte glycoprotein (Lisowska E. 2001 315:366 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 36 theme human 393:397 arg1 glycophorins 399:410 human glycophorins 393:410 human glycophorins 393:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 0 37 theme C 30:30 arg1 N-glycans 32:40 glycophorin C N-glycans 18:40 glycophorin C N-glycans 18:40 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 0 37 theme C 30:30 arg1 component 54:62 a putative component 43:62 a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand 43:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 6 38 theme Plasmodium 816:825 arg1 falciparum 827:836 Plasmodium falciparum EBA-140 816:844 Plasmodium falciparum EBA-140 merozoite ligand 816:861 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 11 39 theme Gerbich 1619:1625 arg1 variant 1627:1633 the GPC Gerbich variant 1611:1633 the GPC Gerbich variant 1611:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 2 40 theme erythrocyte 325:335 arg1 properties 379:388 Antigenic properties 369:388 Antigenic properties of human glycophorins 369:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 40 theme erythrocyte 325:335 arg1 glycoprotein 337:348 the major erythrocyte glycoprotein 315:348 the major erythrocyte glycoprotein (Lisowska E. 2001 315:366 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 8 41 theme many 1084:1087 arg1 N-glycans 1093:1101 many GPC N-glycans 1084:1101 many GPC N-glycans 1084:1101 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 11 42 theme lactosamine 1590:1600 arg1 units 1602:1606 repeating lactosamine units 1580:1606 repeating lactosamine units of the GPC Gerbich variant 1580:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 7 43 theme structures 889:898 arg1 elucidation 868:878 The elucidation 864:878 The elucidation of these structures 864:898 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 1 44 theme red 213:215 arg1 membranes 228:236 human red blood cell membranes 207:236 human red blood cell membranes carrying the Gerbich blood group antigens 207:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 45 contain contain 1145:1151 arg2 structures 1169:1178 polylactosamine structures 1153:1178 polylactosamine structures capped with fucose 1153:1197 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 8 45 contain contain 1145:1151 arg1 several 1137:1143 several 1137:1143 several 1137:1143 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 11 46 with interaction 1753:1763 arg1 GPC 1770:1772 GPC 1770:1772 GPC 1770:1772 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 1 47 theme cell 223:226 arg1 membranes 228:236 human red blood cell membranes 207:236 human red blood cell membranes carrying the Gerbich blood group antigens 207:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 48 theme H2 1111:1112 arg1 structures 1122:1131 H2 antigen structures 1111:1131 H2 antigen structures 1111:1131 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 1 49 gly glycosylated 163:174 arg1 sialoglycoproteins 185:202 highly glycosylated integral sialoglycoproteins 156:202 highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens 156:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 49 gly glycosylated 163:174 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 11 50 theme chains 1562:1567 arg1 fucosylation 1533:1544 preferable terminal fucosylation 1513:1544 preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1513:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 50 theme chains 1562:1567 arg1 explanation 1647:1657 an explanation 1644:1657 an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure 1644:1826 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 1 51 theme Gerbich 251:257 arg1 antigens 271:278 the Gerbich blood group antigens 247:278 the Gerbich blood group antigens 247:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 52 gly sialoglycoproteins 185:202 arg1 sialoglycoproteins 185:202 highly glycosylated integral sialoglycoproteins 156:202 highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens 156:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 52 gly sialoglycoproteins 185:202 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 53 theme polylactosamine 1153:1167 arg1 structures 1169:1178 polylactosamine structures 1153:1178 polylactosamine structures capped with fucose 1153:1197 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 11 54 theme N-glycan 1809:1816 arg1 structure 1818:1826 the GPC N-glycan structure 1801:1826 the GPC N-glycan structure 1801:1826 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 0 55 theme falciparum 104:113 arg1 ligand 123:128 Plasmodium falciparum EBA-140 ligand 93:128 Plasmodium falciparum EBA-140 ligand 93:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 10 56 theme interpretation 1403:1416 arg1 possibility 1384:1394 a possibility 1382:1394 a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC 1382:1490 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 3 57 theme oligosaccharide 512:526 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 57 theme oligosaccharide 512:526 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 57 theme oligosaccharide 512:526 arg1 sites 539:543 oligosaccharide attachment sites 512:543 oligosaccharide attachment sites 512:543 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 1 58 theme integral 176:183 arg1 sialoglycoproteins 185:202 highly glycosylated integral sialoglycoproteins 156:202 highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens 156:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 58 theme integral 176:183 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 59 contain contain 1103:1109 arg1 N-glycans 1093:1101 many GPC N-glycans 1084:1101 many GPC N-glycans 1084:1101 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 8 59 contain contain 1103:1109 arg2 structures 1122:1131 H2 antigen structures 1111:1131 H2 antigen structures 1111:1131 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 3 60 theme glycophorin 566:576 arg1 sequence 499:506 Amino-acid sequence 488:506 Amino-acid sequence 488:506 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 60 theme glycophorin 566:576 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 60 theme glycophorin 566:576 arg1 sites 539:543 oligosaccharide attachment sites 512:543 oligosaccharide attachment sites 512:543 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 60 theme glycophorin 566:576 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 61 theme human 548:552 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 5 62 theme GPC 662:664 arg1 N-glycans 666:674 GPC N-glycans 662:674 GPC N-glycans 662:674 are well characterized but the structure of GPC N-glycans has remained unknown. 25552259 2 63 dep O- 285:286 arg1 The 281:283 The 281:283 The 281:283 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 63 dep O- 285:286 arg1 update 417:422 an update 414:422 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 281:423 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 11 64 theme repeating 1580:1588 arg1 units 1602:1606 repeating lactosamine units 1580:1606 repeating lactosamine units of the GPC Gerbich variant 1580:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 9 65 theme N-glycans 1266:1274 arg1 heterogeneity 1241:1253 structural heterogeneity 1230:1253 structural heterogeneity of the GPC N-glycans 1230:1274 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 11 66 theme preferable 1513:1522 arg1 fucosylation 1533:1544 preferable terminal fucosylation 1513:1544 preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1513:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 66 theme preferable 1513:1522 arg1 explanation 1647:1657 an explanation 1644:1657 an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure 1644:1826 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 67 theme EBA-140 1745:1751 arg1 interaction 1753:1763 the EBA-140 interaction 1741:1763 the EBA-140 interaction with GPC 1741:1772 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 67 theme EBA-140 1745:1751 arg1 dependent 1788:1796 dependent 1788:1796 dependent 1788:1796 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 7 68 theme site 961:964 arg1 characterization 925:940 full characterization 920:940 full characterization of the GPC binding site for the EBA-140 ligand 920:987 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 11 69 dep indication 1725:1734 arg1 interaction 1753:1763 the EBA-140 interaction 1741:1763 the EBA-140 interaction with GPC 1741:1772 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 69 dep indication 1725:1734 arg1 dependent 1788:1796 dependent 1788:1796 dependent 1788:1796 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 0 70 theme Plasmodium 93:102 arg1 falciparum 104:113 Plasmodium falciparum EBA-140 93:121 Plasmodium falciparum EBA-140 ligand 93:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 0 71 theme receptor 75:82 arg1 site 84:87 the GPC receptor site 67:87 the GPC receptor site for Plasmodium falciparum EBA-140 ligand 67:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 9 72 theme structural 1302:1311 arg1 elements 1313:1320 structural elements 1302:1320 structural elements not found in glycophorin A N-glycans 1302:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 8 73 theme structural 1016:1025 arg1 analysis 1027:1034 detailed structural analysis 1007:1034 detailed structural analysis using sequential mass spectrometry 1007:1069 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 7 74 theme GPC 949:951 arg1 site 961:964 the GPC binding site 945:964 the GPC binding site for the EBA-140 ligand 945:987 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 2 75 dep glycoprotein 337:348 arg1 E. 360:361 E. 360:361 E. 360:361 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 3 76 dep Tomita 456:461 arg1 sequence 499:506 Amino-acid sequence 488:506 Amino-acid sequence 488:506 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 76 dep Tomita 456:461 arg1 sites 539:543 oligosaccharide attachment sites 512:543 oligosaccharide attachment sites 512:543 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 76 dep Tomita 456:461 arg1 M 463:463 M 463:463 M 463:463 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 76 dep Tomita 456:461 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 76 dep Tomita 456:461 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 10 77 theme EBA-140 1470:1476 arg1 ligand 1478:1483 P. falciparum EBA-140 ligand 1456:1483 P. falciparum EBA-140 ligand 1456:1483 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 2 78 theme Antigenic 369:377 arg1 properties 379:388 Antigenic properties 369:388 Antigenic properties of human glycophorins 369:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 78 theme Antigenic 369:377 arg1 glycoprotein 337:348 the major erythrocyte glycoprotein 315:348 the major erythrocyte glycoprotein (Lisowska E. 2001 315:366 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 8 79 theme mass 1053:1056 arg1 spectrometry 1058:1069 sequential mass spectrometry 1042:1069 sequential mass spectrometry 1042:1069 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 7 80 theme full 920:923 arg1 characterization 925:940 full characterization 920:940 full characterization of the GPC binding site for the EBA-140 ligand 920:987 The elucidation of these structures seems essential for full characterization of the GPC binding site for the EBA-140 ligand. 25552259 11 81 theme N-glycosidic 1549:1560 arg1 chains 1562:1567 N-glycosidic chains 1549:1567 N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1549:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 10 82 theme P. 1456:1457 arg1 ligand 1478:1483 P. falciparum EBA-140 ligand 1456:1483 P. falciparum EBA-140 ligand 1456:1483 Our results also open a possibility of new interpretation of the data concerning the binding of P. falciparum EBA-140 ligand to GPC. 25552259 0 83 theme glycophorin 18:28 arg1 N-glycans 32:40 glycophorin C N-glycans 18:40 glycophorin C N-glycans 18:40 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 0 83 theme glycophorin 18:28 arg1 component 54:62 a putative component 43:62 a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand 43:128 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 9 84 theme A 1347:1347 arg1 N-glycans 1349:1357 glycophorin A N-glycans 1335:1357 glycophorin A N-glycans 1335:1357 The results obtained indicate structural heterogeneity of the GPC N-glycans and show the existence of structural elements not found in glycophorin A N-glycans. 25552259 11 85 theme GPC 1706:1708 arg1 Gerbich 1710:1716 GPC Gerbich 1706:1716 GPC Gerbich 1706:1716 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 1 86 dep Glycophorins 131:142 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 86 dep Glycophorins 131:142 arg1 D 150:150 D 150:150 D 150:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 86 dep Glycophorins 131:142 arg1 C 144:144 C 144:144 C 144:144 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 2 87 theme glycophorins 399:410 arg1 properties 379:388 Antigenic properties 369:388 Antigenic properties of human glycophorins 369:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 87 theme glycophorins 399:410 arg1 glycoprotein 337:348 the major erythrocyte glycoprotein 315:348 the major erythrocyte glycoprotein (Lisowska E. 2001 315:366 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 0 88 theme N-glycans 32:40 arg1 structures 4:13 The structures 0:13 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 0:129 The structures of glycophorin C N-glycans, a putative component of the GPC receptor site for Plasmodium falciparum EBA-140 ligand. 25552259 6 89 theme falciparum 827:836 arg1 ligand 856:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 6 90 with interaction 799:809 arg1 ligand 856:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 Plasmodium falciparum EBA-140 merozoite ligand 816:861 This problem became important since it was reported that GPC N-glycans play an essential role in the interaction with Plasmodium falciparum EBA-140 merozoite ligand. 25552259 11 91 theme GPC 1615:1617 arg1 variant 1627:1633 the GPC Gerbich variant 1611:1633 the GPC Gerbich variant 1611:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 11 92 gly fucosylation 1533:1544 arg1 chains 1562:1567 N-glycosidic chains 1549:1567 N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant 1549:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 8 93 theme GPC 1089:1091 arg1 N-glycans 1093:1101 many GPC N-glycans 1084:1101 many GPC N-glycans 1084:1101 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 3 94 theme Marchesi 469:476 arg1 sequence 499:506 Amino-acid sequence 488:506 Amino-acid sequence 488:506 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 94 theme Marchesi 469:476 arg1 sites 539:543 oligosaccharide attachment sites 512:543 oligosaccharide attachment sites 512:543 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 94 theme Marchesi 469:476 arg1 VT. 478:480 Marchesi VT. 1975 469:485 Marchesi VT. 1975 469:485 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 3 94 theme Marchesi 469:476 arg1 glycophorin 566:576 human erythrocyte glycophorin 548:576 human erythrocyte glycophorin 548:576 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 2 95 theme major 319:323 arg1 properties 379:388 Antigenic properties 369:388 Antigenic properties of human glycophorins 369:410 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 2 95 theme major 319:323 arg1 glycoprotein 337:348 the major erythrocyte glycoprotein 315:348 the major erythrocyte glycoprotein (Lisowska E. 2001 315:366 The O- and N-glycosidic chains of the major erythrocyte glycoprotein (Lisowska E. 2001, Antigenic properties of human glycophorins - an update. 25552259 1 96 theme blood 217:221 arg1 membranes 228:236 human red blood cell membranes 207:236 human red blood cell membranes carrying the Gerbich blood group antigens 207:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 8 97 theme antigen 1114:1120 arg1 structures 1122:1131 H2 antigen structures 1111:1131 H2 antigen structures 1111:1131 We have employed detailed structural analysis using sequential mass spectrometry to show that many GPC N-glycans contain H2 antigen structures and several contain polylactosamine structures capped with fucose. 25552259 3 98 dep Biol 437:440 arg1 Tomita 456:461 Tomita 456:461 Tomita 456:461 Adv Exp Med Biol, 491:155-169; Tomita M and Marchesi VT. 1975, Amino-acid sequence and oligosaccharide attachment sites of human erythrocyte glycophorin. 25552259 11 99 theme variant 1627:1633 arg1 units 1602:1606 repeating lactosamine units 1580:1606 repeating lactosamine units of the GPC Gerbich variant 1580:1633 We hypothesize that preferable terminal fucosylation of N-glycosidic chains containing repeating lactosamine units of the GPC Gerbich variant could be an explanation for why the EBA-140 ligand does not react with GPC Gerbich and an indication that the EBA-140 interaction with GPC is distinctly dependent on the GPC N-glycan structure. 25552259 1 100 theme membranes 228:236 arg1 sialoglycoproteins 185:202 highly glycosylated integral sialoglycoproteins 156:202 highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens 156:278 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 25552259 1 100 theme membranes 228:236 arg1 Glycophorins 131:142 Glycophorins C and D 131:150 Glycophorins C and D 131:150 Glycophorins C and D are highly glycosylated integral sialoglycoproteins of human red blood cell membranes carrying the Gerbich blood group antigens. 27288408 0 0 theme Relaxed 99:105 arg1 Specificity 117:127 Remarkably Relaxed Substrate Specificity 88:127 Remarkably Relaxed Substrate Specificity 88:127 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 3 1 from mutagenesis 629:639 arg1 pyogenes 682:689 Streptococcus pyogenes 668:689 Streptococcus pyogenes of serotype M49 (Endo-S2) 668:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 2 theme relaxed 1322:1328 arg1 specificity 1340:1350 remarkably relaxed substrate specificity 1311:1350 remarkably relaxed substrate specificity 1311:1350 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 3 contain have 1306:1309 arg1 glycosynthases 1234:1247 the Endo-S2 glycosynthases 1222:1247 the Endo-S2 glycosynthases described here 1222:1262 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 3 contain have 1306:1309 arg1 capable 1361:1367 capable 1361:1367 capable 1361:1367 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 3 contain have 1306:1309 arg2 specificity 1340:1350 remarkably relaxed substrate specificity 1311:1350 remarkably relaxed substrate specificity 1311:1350 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 1 4 theme profound 194:201 arg1 impact 203:208 a profound impact 192:208 a profound impact 192:208 Glycosylation can exert a profound impact on the structures and biological functions of antibodies. 27288408 5 5 theme substrate 1330:1338 arg1 specificity 1340:1350 remarkably relaxed substrate specificity 1311:1350 remarkably relaxed substrate specificity 1311:1350 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 3 6 from evaluation 725:734 arg1 pyogenes 682:689 Streptococcus pyogenes 668:689 Streptococcus pyogenes of serotype M49 (Endo-S2) 668:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 7 theme transglycosylation 786:803 arg1 activities 805:814 their hydrolysis and transglycosylation activities 765:814 activities 805:814 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 8 theme major 1391:1395 arg1 types 1397:1401 three major types 1385:1401 three major types (complex, high-mannose, and hybrid type) of N-glycans 1385:1455 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 8 theme major 1391:1395 arg1 type 1438:1441 hybrid type 1431:1441 hybrid type 1431:1441 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 8 theme major 1391:1395 arg1 high-mannose 1413:1424 high-mannose 1413:1424 high-mannose 1413:1424 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 8 theme major 1391:1395 arg1 complex 1404:1410 complex 1404:1410 complex 1404:1410 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 7 9 theme mutants 1682:1688 arg1 usefulness 1640:1649 The usefulness 1636:1649 The usefulness of these Endo-S2 glycosynthase mutants 1636:1688 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 0 10 theme Substrate 107:115 arg1 Specificity 117:127 Remarkably Relaxed Substrate Specificity 88:127 Remarkably Relaxed Substrate Specificity 88:127 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 4 11 contain possessed 936:944 arg2 activity 972:979 potent transglycosylation activity 946:979 potent transglycosylation activity 946:979 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 4 11 contain possessed 936:944 arg1 some 931:934 some 931:934 some 931:934 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 0 12 theme Antibody 133:140 arg1 Remodeling 156:165 Antibody Glycosylation Remodeling 133:165 Antibody Glycosylation Remodeling 133:165 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 5 13 theme Endo-S2 1226:1232 arg1 glycosynthases 1234:1247 the Endo-S2 glycosynthases 1222:1247 the Endo-S2 glycosynthases described here 1222:1262 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 13 theme Endo-S2 1226:1232 arg1 capable 1361:1367 capable 1361:1367 capable 1361:1367 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 2 14 theme method 496:501 arg1 application 476:486 the broad application 466:486 the broad application of this method 466:501 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 2 15 theme transglycosylation 349:366 arg1 approach 368:375 the endoglycosidase-catalyzed deglycosylation and transglycosylation approach 299:375 approach 368:375 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 5 16 theme N-glycans 1447:1455 arg1 types 1397:1401 three major types 1385:1401 three major types (complex, high-mannose, and hybrid type) of N-glycans 1385:1455 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 16 theme N-glycans 1447:1455 arg1 type 1438:1441 hybrid type 1431:1441 hybrid type 1431:1441 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 16 theme N-glycans 1447:1455 arg1 high-mannose 1413:1424 high-mannose 1413:1424 high-mannose 1413:1424 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 16 theme N-glycans 1447:1455 arg1 complex 1404:1410 complex 1404:1410 complex 1404:1410 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 6 17 theme glycosynthase 1521:1533 arg1 active 1570:1575 active 1570:1575 active 1570:1575 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 6 17 theme glycosynthase 1521:1533 arg1 mutants 1535:1541 the Endo-S2 glycosynthase mutants 1509:1541 the Endo-S2 glycosynthase mutants 1509:1541 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 7 18 theme Endo-S2 1660:1666 arg1 mutants 1682:1688 these Endo-S2 glycosynthase mutants 1654:1688 these Endo-S2 glycosynthase mutants 1654:1688 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 4 19 from residue 856:862 arg1 mutations 831:839 mutations 831:839 mutations at the Asp-184 residue 831:862 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 6 20 from general 1580:1586 arg1 active 1570:1575 active 1570:1575 active 1570:1575 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 6 20 from general 1580:1586 arg1 mutants 1535:1541 the Endo-S2 glycosynthase mutants 1509:1541 the Endo-S2 glycosynthase mutants 1509:1541 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 2 21 theme broad 470:474 arg1 application 476:486 the broad application 466:486 the broad application of this method 466:501 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 4 22 theme hydrolysis 997:1006 arg1 activity 1008:1015 diminished hydrolysis activity 986:1015 diminished hydrolysis activity 986:1015 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 5 23 theme antibody 1461:1468 arg1 remodeling 1484:1493 antibody glycosylation remodeling 1461:1493 antibody glycosylation remodeling 1461:1493 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 2 24 theme promising 394:402 arg1 platform 404:411 a promising platform 392:411 a promising platform to produce homogeneous glycoforms of antibodies 392:459 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 3 25 theme site-directed 615:627 arg1 mutagenesis 629:639 a systematic site-directed mutagenesis 602:639 a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 602:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 0 26 theme Glycosynthase 0:12 arg1 Mutants 14:20 Glycosynthase Mutants 0:20 Glycosynthase Mutants of Endoglycosidase S2 0:42 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 4 27 theme diminished 986:995 arg1 activity 1008:1015 diminished hydrolysis activity 986:1015 diminished hydrolysis activity 986:1015 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 7 28 theme glycosynthase 1668:1680 arg1 mutants 1682:1688 these Endo-S2 glycosynthase mutants 1654:1688 these Endo-S2 glycosynthase mutants 1654:1688 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 5 29 theme type 1206:1209 arg1 N-glycans 1211:1219 complex type N-glycans 1198:1219 complex type N-glycans 1198:1219 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 0 30 theme Endoglycosidase 25:39 arg1 S2 41:42 Endoglycosidase S2 25:42 Endoglycosidase S2 25:42 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 4 31 theme potent 946:951 arg1 activity 972:979 potent transglycosylation activity 946:979 potent transglycosylation activity 946:979 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 3 32 theme endoglycosidase 647:661 arg1 evaluation 725:734 the evaluation 721:734 the evaluation of the resulting mutants 721:759 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 32 theme endoglycosidase 647:661 arg1 mutagenesis 629:639 a systematic site-directed mutagenesis 602:639 a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 602:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 33 theme Endo-S 1153:1158 arg1 mutants 1160:1166 the previously reported Endo-S mutants 1129:1166 the previously reported Endo-S mutants that are limited to action on complex type N-glycans 1129:1219 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 33 theme Endo-S 1153:1158 arg1 limited 1177:1183 limited 1177:1183 limited 1177:1183 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 6 34 theme Endo-S2 1513:1519 arg1 active 1570:1575 active 1570:1575 active 1570:1575 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 6 34 theme Endo-S2 1513:1519 arg1 mutants 1535:1541 the Endo-S2 glycosynthase mutants 1509:1541 the Endo-S2 glycosynthase mutants 1509:1541 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 7 35 theme antibodies 1777:1786 arg1 remodeling 1736:1745 an efficient glycosylation remodeling 1709:1745 an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin) 1709:1825 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 0 36 theme Glycosylation 142:154 arg1 Remodeling 156:165 Antibody Glycosylation Remodeling 133:165 Antibody Glycosylation Remodeling 133:165 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 5 37 theme complex 1198:1204 arg1 N-glycans 1211:1219 complex type N-glycans 1198:1219 complex type N-glycans 1198:1219 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 4 38 theme Asp-184 848:854 arg1 residue 856:862 the Asp-184 residue 844:862 the Asp-184 residue 844:862 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 7 39 theme glycosylation 1722:1734 arg1 remodeling 1736:1745 an efficient glycosylation remodeling 1709:1745 an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin) 1709:1825 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 1 40 theme biological 232:241 arg1 functions 243:251 biological functions 232:251 biological functions 232:251 Glycosylation can exert a profound impact on the structures and biological functions of antibodies. 27288408 2 41 theme mutants 567:573 arg1 availability 520:531 the availability 516:531 the availability of highly efficient glycosynthase mutants 516:573 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 2 42 gly glycoforms 436:445 arg1 antibodies 450:459 antibodies 450:459 antibodies 450:459 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 3 43 theme resulting 743:751 arg1 mutants 753:759 the resulting mutants 739:759 the resulting mutants 739:759 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 44 theme M49 703:705 arg1 pyogenes 682:689 Streptococcus pyogenes 668:689 Streptococcus pyogenes of serotype M49 (Endo-S2) 668:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 45 dep mutants 1160:1166 arg1 contrast 1117:1124 contrast 1117:1124 contrast 1117:1124 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 0 46 theme S2 41:42 arg1 Mutants 14:20 Glycosynthase Mutants 0:20 Glycosynthase Mutants of Endoglycosidase S2 0:42 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 7 47 theme therapeutic 1754:1764 arg1 trastuzumab 1803:1813 trastuzumab 1803:1813 trastuzumab (Herceptin) 1803:1825 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 7 47 theme therapeutic 1754:1764 arg1 rituximab 1789:1797 rituximab 1789:1797 rituximab 1789:1797 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 7 47 theme therapeutic 1754:1764 arg1 antibodies 1777:1786 two therapeutic monoclonal antibodies 1750:1786 two therapeutic monoclonal antibodies 1750:1786 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 2 48 theme glycosynthase 553:565 arg1 mutants 567:573 highly efficient glycosynthase mutants 536:573 highly efficient glycosynthase mutants 536:573 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 1 49 dep structures 217:226 arg1 the 213:215 the 213:215 the 213:215 Glycosylation can exert a profound impact on the structures and biological functions of antibodies. 27288408 0 50 theme Transglycosylation 56:73 arg1 Activity 75:82 Potent Transglycosylation Activity 49:82 Potent Transglycosylation Activity 49:82 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 7 51 theme monoclonal 1766:1775 arg1 trastuzumab 1803:1813 trastuzumab 1803:1813 trastuzumab (Herceptin) 1803:1825 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 7 51 theme monoclonal 1766:1775 arg1 rituximab 1789:1797 rituximab 1789:1797 rituximab 1789:1797 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 7 51 theme monoclonal 1766:1775 arg1 antibodies 1777:1786 two therapeutic monoclonal antibodies 1750:1786 two therapeutic monoclonal antibodies 1750:1786 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 27288408 4 52 theme transglycosylation 953:970 arg1 activity 972:979 potent transglycosylation activity 946:979 potent transglycosylation activity 946:979 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 6 53 located found 1548:1552 arg2 active 1570:1575 active 1570:1575 active 1570:1575 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 6 53 located found 1548:1552 arg2 mutants 1535:1541 the Endo-S2 glycosynthase mutants 1509:1541 the Endo-S2 glycosynthase mutants 1509:1541 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 6 53 located found 1548:1552 arg1 addition 1499:1506 addition 1499:1506 addition 1499:1506 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 2 54 theme homogeneous 424:434 arg1 glycoforms 436:445 homogeneous glycoforms 424:445 homogeneous glycoforms of antibodies 424:459 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 0 55 theme Potent 49:54 arg1 Activity 75:82 Potent Transglycosylation Activity 49:82 Potent Transglycosylation Activity 49:82 Glycosynthase Mutants of Endoglycosidase S2 Show Potent Transglycosylation Activity and Remarkably Relaxed Substrate Specificity for Antibody Glycosylation Remodeling. 27288408 3 56 theme serotype 694:701 arg1 M49 703:705 serotype M49 694:705 serotype M49 (Endo-S2) 694:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 56 theme serotype 694:701 arg1 Endo-S2 708:714 Endo-S2 708:714 Endo-S2 708:714 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 57 theme hybrid 1431:1436 arg1 type 1438:1441 hybrid type 1431:1441 hybrid type 1431:1441 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 57 theme hybrid 1431:1436 arg1 types 1397:1401 three major types 1385:1401 three major types (complex, high-mannose, and hybrid type) of N-glycans 1385:1455 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 4 58 theme comparative 1095:1105 arg1 study 1107:1111 the comparative study 1091:1111 the comparative study 1091:1111 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 3 59 theme Streptococcus 668:680 arg1 pyogenes 682:689 Streptococcus pyogenes 668:689 Streptococcus pyogenes of serotype M49 (Endo-S2) 668:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 5 60 dep types 1397:1401 arg1 type 1438:1441 hybrid type 1431:1441 hybrid type 1431:1441 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 60 dep types 1397:1401 arg1 types 1397:1401 three major types 1385:1401 three major types (complex, high-mannose, and hybrid type) of N-glycans 1385:1455 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 60 dep types 1397:1401 arg1 high-mannose 1413:1424 high-mannose 1413:1424 high-mannose 1413:1424 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 60 dep types 1397:1401 arg1 complex 1404:1410 complex 1404:1410 complex 1404:1410 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 4 61 theme different 909:917 arg1 properties 919:928 significantly different properties 895:928 significantly different properties 895:928 We found that mutations at the Asp-184 residue gave mutants that demonstrated significantly different properties, some possessed potent transglycosylation activity with diminished hydrolysis activity but others did not, which would be otherwise difficult to predict without the comparative study. 27288408 2 62 theme Glycosylation 268:280 arg1 remodeling 282:291 Glycosylation remodeling 268:291 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach 268:375 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 1 63 theme antibodies 256:265 arg1 structures 217:226 structures 217:226 structures 217:226 Glycosylation can exert a profound impact on the structures and biological functions of antibodies. 27288408 1 63 theme antibodies 256:265 arg1 functions 243:251 biological functions 232:251 biological functions 232:251 Glycosylation can exert a profound impact on the structures and biological functions of antibodies. 27288408 3 64 theme mutants 753:759 arg1 evaluation 725:734 the evaluation 721:734 the evaluation of the resulting mutants 721:759 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 64 theme mutants 753:759 arg1 mutagenesis 629:639 a systematic site-directed mutagenesis 602:639 a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 602:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 2 65 theme endoglycosidase-catalyzed 303:327 arg1 deglycosylation 329:343 the endoglycosidase-catalyzed deglycosylation and transglycosylation approach 299:375 deglycosylation 329:343 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 5 66 theme glycosylation 1470:1482 arg1 remodeling 1484:1493 antibody glycosylation remodeling 1461:1493 antibody glycosylation remodeling 1461:1493 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 3 67 theme systematic 604:613 arg1 mutagenesis 629:639 a systematic site-directed mutagenesis 602:639 a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 602:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 6 68 theme Endo-S 1597:1602 arg1 mutants 1604:1610 the Endo-S mutants 1593:1610 the Endo-S mutants for transglycosylation 1593:1633 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 5 69 from action 1188:1193 arg1 N-glycans 1211:1219 complex type N-glycans 1198:1219 complex type N-glycans 1198:1219 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 6 70 from active 1570:1575 arg1 general 1580:1586 general 1580:1586 general 1580:1586 In addition, the Endo-S2 glycosynthase mutants were found to be much more active in general than the Endo-S mutants for transglycosylation. 27288408 3 71 from pyogenes 682:689 arg1 evaluation 725:734 the evaluation 721:734 the evaluation of the resulting mutants 721:759 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 71 from pyogenes 682:689 arg1 mutagenesis 629:639 a systematic site-directed mutagenesis 602:639 a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 602:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 3 71 from pyogenes 682:689 arg1 endoglycosidase 647:661 an endoglycosidase 644:661 an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) 644:715 We describe in this paper a systematic site-directed mutagenesis of an endoglycosidase from Streptococcus pyogenes of serotype M49 (Endo-S2) and the evaluation of the resulting mutants for their hydrolysis and transglycosylation activities. 27288408 2 72 theme efficient 543:551 arg1 mutants 567:573 highly efficient glycosynthase mutants 536:573 highly efficient glycosynthase mutants 536:573 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 5 73 theme reported 1144:1151 arg1 mutants 1160:1166 the previously reported Endo-S mutants 1129:1166 the previously reported Endo-S mutants that are limited to action on complex type N-glycans 1129:1219 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 5 73 theme reported 1144:1151 arg1 limited 1177:1183 limited 1177:1183 limited 1177:1183 In contrast to the previously reported Endo-S mutants that are limited to action on complex type N-glycans, the Endo-S2 glycosynthases described here, including D184M and D184Q, were found to have remarkably relaxed substrate specificity and were capable of transferring three major types (complex, high-mannose, and hybrid type) of N-glycans for antibody glycosylation remodeling. 27288408 2 74 theme antibodies 450:459 arg1 glycoforms 436:445 homogeneous glycoforms 424:445 homogeneous glycoforms of antibodies 424:459 Glycosylation remodeling using the endoglycosidase-catalyzed deglycosylation and transglycosylation approach is emerging as a promising platform to produce homogeneous glycoforms of antibodies, but the broad application of this method will require the availability of highly efficient glycosynthase mutants. 27288408 7 75 theme efficient 1712:1720 arg1 remodeling 1736:1745 an efficient glycosylation remodeling 1709:1745 an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin) 1709:1825 The usefulness of these Endo-S2 glycosynthase mutants was exemplified by an efficient glycosylation remodeling of two therapeutic monoclonal antibodies, rituximab and trastuzumab (Herceptin). 24623192 3 0 theme protein 362:368 arg1 glycosylation 370:382 protein glycosylation 362:382 protein glycosylation 362:382 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 4 1 theme cell 533:536 arg1 surfaces 538:545 cell surfaces 533:545 cell surfaces 533:545 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 6 2 theme bioorthogonal 776:788 arg1 azides 807:812 other classic bioorthogonal motifs-including azides 762:812 other classic bioorthogonal motifs-including azides 762:812 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 3 3 theme glycosylation 370:382 arg1 scaffolds 323:331 existing cyclopropene scaffolds 301:331 existing cyclopropene scaffolds 301:331 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 3 3 theme glycosylation 370:382 arg1 reporters 349:357 inefficient reporters 337:357 inefficient reporters of protein glycosylation 337:382 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 4 4 theme isolated 551:558 arg1 proteins 560:567 isolated proteins 551:567 isolated proteins 551:567 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 5 5 theme biosynthetic 648:659 arg1 pathways 661:668 cellular biosynthetic pathways 639:668 cellular biosynthetic pathways 639:668 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 3 6 theme branched 400:407 arg1 structures 409:418 their branched structures 394:418 their branched structures 394:418 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 6 7 theme multiple 837:844 arg1 biomolecules 846:857 multiple biomolecules 837:857 multiple biomolecules 837:857 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 6 8 theme classic 768:774 arg1 azides 807:812 other classic bioorthogonal motifs-including azides 762:812 other classic bioorthogonal motifs-including azides 762:812 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 5 9 theme rapid 682:686 arg1 rates 702:706 rapid cycloaddition rates 682:706 rapid cycloaddition rates 682:706 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 6 10 used used 742:745 arg2 probes 728:733 these probes 722:733 these probes 722:733 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 6 11 theme other 762:766 arg1 azides 807:812 other classic bioorthogonal motifs-including azides 762:812 other classic bioorthogonal motifs-including azides 762:812 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 2 12 theme strained 152:159 arg1 rings 161:165 These strained rings 146:165 These strained rings 146:165 These strained rings can be metabolically introduced into target biomolecules and covalently modified via mild cycloaddition chemistries. 24623192 5 13 theme cycloaddition 688:700 arg1 rates 702:706 rapid cycloaddition rates 682:706 rapid cycloaddition rates 682:706 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 0 14 theme cyclopropene 9:20 arg1 reporters 22:30 cyclopropene reporters 9:30 cyclopropene reporters 9:30 Improved cyclopropene reporters for probing protein glycosylation. 24623192 4 15 theme glycans 522:528 arg1 detection 509:517 the robust detection 498:517 the robust detection of glycans on cell surfaces and isolated proteins 498:567 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 4 16 from detection 509:517 arg1 surfaces 538:545 cell surfaces 533:545 cell surfaces 533:545 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 4 16 from detection 509:517 arg1 proteins 560:567 isolated proteins 551:567 isolated proteins 551:567 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 1 17 theme new 99:101 arg1 class 103:107 a new class 97:107 a new class of bioorthogonal chemical reporters 97:143 Cyclopropenes have emerged as a new class of bioorthogonal chemical reporters. 24623192 3 18 theme cyclopropene 310:321 arg1 scaffolds 323:331 existing cyclopropene scaffolds 301:331 existing cyclopropene scaffolds 301:331 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 3 18 theme cyclopropene 310:321 arg1 reporters 349:357 inefficient reporters 337:357 inefficient reporters of protein glycosylation 337:382 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 5 19 theme cellular 639:646 arg1 pathways 661:668 cellular biosynthetic pathways 639:668 cellular biosynthetic pathways 639:668 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 3 20 theme sluggish 424:431 arg1 rates 433:437 sluggish rates 424:437 sluggish rates of reactivity 424:451 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 6 21 with tandem 750:755 arg1 azides 807:812 other classic bioorthogonal motifs-including azides 762:812 other classic bioorthogonal motifs-including azides 762:812 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 24623192 0 22 theme protein 44:50 arg1 glycosylation 52:64 protein glycosylation 44:64 protein glycosylation 44:64 Improved cyclopropene reporters for probing protein glycosylation. 24623192 5 23 with compatible 623:632 arg1 pathways 661:668 cellular biosynthetic pathways 639:668 cellular biosynthetic pathways 639:668 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 4 24 theme cyclopropenes 480:492 arg1 set 473:475 a set 471:475 a set of cyclopropenes 471:492 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 4 24 theme cyclopropenes 480:492 arg1 cyclopropenes 480:492 cyclopropenes 480:492 cyclopropenes 480:492 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 2 25 theme cycloaddition 257:269 arg1 chemistries 271:281 mild cycloaddition chemistries 252:281 mild cycloaddition chemistries 252:281 These strained rings can be metabolically introduced into target biomolecules and covalently modified via mild cycloaddition chemistries. 24623192 3 26 theme existing 301:308 arg1 scaffolds 323:331 existing cyclopropene scaffolds 301:331 existing cyclopropene scaffolds 301:331 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 3 26 theme existing 301:308 arg1 reporters 349:357 inefficient reporters 337:357 inefficient reporters of protein glycosylation 337:382 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 4 27 theme robust 502:507 arg1 detection 509:517 the robust detection 498:517 the robust detection of glycans on cell surfaces and isolated proteins 498:567 Here we describe a set of cyclopropenes for the robust detection of glycans on cell surfaces and isolated proteins. 24623192 2 28 theme mild 252:255 arg1 chemistries 271:281 mild cycloaddition chemistries 252:281 mild cycloaddition chemistries 252:281 These strained rings can be metabolically introduced into target biomolecules and covalently modified via mild cycloaddition chemistries. 24623192 3 29 theme reactivity 442:451 arg1 rates 433:437 sluggish rates 424:437 sluggish rates of reactivity 424:451 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 3 29 theme reactivity 442:451 arg1 structures 409:418 their branched structures 394:418 their branched structures 394:418 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 5 30 theme carbamate 595:603 arg1 linkages 605:612 carbamate linkages 595:612 carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates 595:706 These scaffolds comprise carbamate linkages that are compatible with cellular biosynthetic pathways and exhibit rapid cycloaddition rates. 24623192 1 31 theme bioorthogonal 112:124 arg1 reporters 135:143 bioorthogonal chemical reporters 112:143 bioorthogonal chemical reporters 112:143 Cyclopropenes have emerged as a new class of bioorthogonal chemical reporters. 24623192 1 32 theme chemical 126:133 arg1 reporters 135:143 bioorthogonal chemical reporters 112:143 bioorthogonal chemical reporters 112:143 Cyclopropenes have emerged as a new class of bioorthogonal chemical reporters. 24623192 1 33 theme reporters 135:143 arg1 class 103:107 a new class 97:107 a new class of bioorthogonal chemical reporters 97:143 Cyclopropenes have emerged as a new class of bioorthogonal chemical reporters. 24623192 3 34 theme inefficient 337:347 arg1 scaffolds 323:331 existing cyclopropene scaffolds 301:331 existing cyclopropene scaffolds 301:331 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 3 34 theme inefficient 337:347 arg1 reporters 349:357 inefficient reporters 337:357 inefficient reporters of protein glycosylation 337:382 While versatile, existing cyclopropene scaffolds are inefficient reporters of protein glycosylation, owing to their branched structures and sluggish rates of reactivity. 24623192 2 35 theme target 204:209 arg1 biomolecules 211:222 target biomolecules 204:222 target biomolecules 204:222 These strained rings can be metabolically introduced into target biomolecules and covalently modified via mild cycloaddition chemistries. 24623192 6 36 theme motifs-including 790:805 arg1 azides 807:812 other classic bioorthogonal motifs-including azides 762:812 other classic bioorthogonal motifs-including azides 762:812 Furthermore, these probes can be used in tandem with other classic bioorthogonal motifs-including azides and alkynes-to examine multiple biomolecules in tandem. 28985062 3 0 theme remote 370:375 arg1 activation 377:386 the remote activation 366:386 the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor 366:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 5 1 dep yielding 727:734 arg1 high 722:725 high 722:725 high 722:725 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 3 2 from N3-site 407:413 arg1 Tf2O 395:398 the Tf2O 391:398 the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor 391:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 7 3 theme oligosaccharide 988:1002 arg1 synthesis 1004:1012 the oligosaccharide synthesis 984:1012 the oligosaccharide synthesis 984:1012 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 5 4 dep mild 716:719 arg1 fast 737:740 fast 737:740 fast 737:740 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 4 dep mild 716:719 arg1 mild 716:719 mild 716:719 mild 716:719 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 4 dep mild 716:719 arg1 yielding 727:734 yielding 727:734 yielding 727:734 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 4 dep mild 716:719 arg1 reaction 704:711 The reaction 700:711 The reaction 700:711 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 4 dep mild 716:719 arg1 suitable 746:753 suitable 746:753 suitable 746:753 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 5 contain containing 766:775 arg2 C2-esters 796:804 C2-esters 796:804 C2-esters 796:804 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 5 contain containing 766:775 arg1 donors 759:764 donors 759:764 donors containing both C2-ethers and C2-esters as well 759:812 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 5 5 contain containing 766:775 arg2 C2-ethers 782:790 C2-ethers 782:790 C2-ethers 782:790 The reaction is mild, high yielding, fast and suitable for donors containing both C2-ethers and C2-esters as well. 28985062 6 6 theme glycosidation 838:850 arg1 strategy 852:859 The regenerative-donor glycosidation strategy 815:859 The regenerative-donor glycosidation strategy 815:859 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 6 6 theme glycosidation 838:850 arg1 promising 864:872 promising 864:872 promising 864:872 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 1 7 theme increased 196:204 arg1 reactivity 206:215 the increased reactivity 192:215 the increased reactivity 192:215 Hydroxybenzotriazole is routinely used in peptide chemistry for reducing racemization due to the increased reactivity. 28985062 6 8 theme regenerative-donor 819:836 arg1 strategy 852:859 The regenerative-donor glycosidation strategy 815:859 The regenerative-donor glycosidation strategy 815:859 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 6 8 theme regenerative-donor 819:836 arg1 promising 864:872 promising 864:872 promising 864:872 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 3 9 theme HOBt 418:421 arg1 HOBt 418:421 HOBt 418:421 HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor 418:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 3 9 theme HOBt 418:421 arg1 N3-site 407:413 the N3-site 403:413 the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor 403:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 6 10 theme further 928:934 arg1 utilization 936:946 further utilization 928:946 further utilization 928:946 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 4 11 theme leaving 593:599 arg1 group 601:605 the leaving group 589:605 the leaving group 589:605 Further, equilibration of the zwitterionic benzotriazolyl species makes the leaving group noncompetitive and generates the nucleofuge that has been reconverted to the glycosyl donor. 28985062 3 12 theme oxocarbenium 456:467 arg1 ion 469:471 the oxocarbenium ion 452:471 the oxocarbenium ion that was attacked by the glycosyl acceptor 452:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 7 13 theme methodology 968:978 arg1 utility 953:959 The utility 949:959 The utility of the methodology for the oligosaccharide synthesis 949:1012 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 6 14 theme glycosyl 909:916 arg1 donor 918:922 the glycosyl donor 905:922 the glycosyl donor for further utilization 905:946 The regenerative-donor glycosidation strategy is promising as it enables us to regenerate the glycosyl donor for further utilization. 28985062 0 15 theme Remote 44:49 arg1 Activation 51:60 the Remote Activation 40:60 the Remote Activation of Hydroxybenzotriazolyl Glycosides 40:96 Nucleofuge Generating Glycosidations by the Remote Activation of Hydroxybenzotriazolyl Glycosides. 28985062 7 16 theme complex 1110:1116 arg1 core 1087:1090 the branched pentamannan core 1062:1090 the branched pentamannan core of the HIV1-gp120 complex 1062:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 3 17 theme ion 469:471 arg1 extrusion 439:447 the extrusion 435:447 the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor 435:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 4 18 theme benzotriazolyl 560:573 arg1 species 575:581 the zwitterionic benzotriazolyl species 543:581 the zwitterionic benzotriazolyl species 543:581 Further, equilibration of the zwitterionic benzotriazolyl species makes the leaving group noncompetitive and generates the nucleofuge that has been reconverted to the glycosyl donor. 28985062 7 19 theme branched 1066:1073 arg1 core 1087:1090 the branched pentamannan core 1062:1090 the branched pentamannan core of the HIV1-gp120 complex 1062:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 4 20 theme glycosyl 684:691 arg1 donor 693:697 the glycosyl donor 680:697 the glycosyl donor 680:697 Further, equilibration of the zwitterionic benzotriazolyl species makes the leaving group noncompetitive and generates the nucleofuge that has been reconverted to the glycosyl donor. 28985062 4 21 theme zwitterionic 547:558 arg1 species 575:581 the zwitterionic benzotriazolyl species 543:581 the zwitterionic benzotriazolyl species 543:581 Further, equilibration of the zwitterionic benzotriazolyl species makes the leaving group noncompetitive and generates the nucleofuge that has been reconverted to the glycosyl donor. 28985062 2 22 theme hydroxybenzotriazolyl 247:267 arg1 glucosides 269:278 very stable hydroxybenzotriazolyl glucosides 235:278 very stable hydroxybenzotriazolyl glucosides 235:278 In this article, very stable hydroxybenzotriazolyl glucosides were identified to undergo glycosidation. 28985062 7 23 theme pentamannan 1075:1085 arg1 core 1087:1090 the branched pentamannan core 1062:1090 the branched pentamannan core of the HIV1-gp120 complex 1062:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 1 24 theme peptide 141:147 arg1 chemistry 149:157 peptide chemistry 141:157 peptide chemistry 141:157 Hydroxybenzotriazole is routinely used in peptide chemistry for reducing racemization due to the increased reactivity. 28985062 7 25 theme HIV1-gp120 1099:1108 arg1 complex 1110:1116 the HIV1-gp120 complex 1095:1116 the HIV1-gp120 complex 1095:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 2 26 theme stable 240:245 arg1 glucosides 269:278 very stable hydroxybenzotriazolyl glucosides 235:278 very stable hydroxybenzotriazolyl glucosides 235:278 In this article, very stable hydroxybenzotriazolyl glucosides were identified to undergo glycosidation. 28985062 1 27 used used 133:136 arg2 Hydroxybenzotriazole 99:118 Hydroxybenzotriazole 99:118 Hydroxybenzotriazole 99:118 Hydroxybenzotriazole is routinely used in peptide chemistry for reducing racemization due to the increased reactivity. 28985062 4 28 theme species 575:581 arg1 equilibration 526:538 equilibration 526:538 equilibration of the zwitterionic benzotriazolyl species 526:581 Further, equilibration of the zwitterionic benzotriazolyl species makes the leaving group noncompetitive and generates the nucleofuge that has been reconverted to the glycosyl donor. 28985062 7 29 theme core 1087:1090 arg1 synthesis 1049:1057 the successful synthesis 1034:1057 the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex 1034:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 3 30 theme glycosyl 498:505 arg1 acceptor 507:514 the glycosyl acceptor 494:514 the glycosyl acceptor 494:514 The reaction was hypothesized to go through the remote activation by the Tf2O at the N3-site of HOBt followed by the extrusion of the oxocarbenium ion that was attacked by the glycosyl acceptor. 28985062 7 31 theme successful 1038:1047 arg1 synthesis 1049:1057 the successful synthesis 1034:1057 the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex 1034:1116 The utility of the methodology for the oligosaccharide synthesis was demonstrated by the successful synthesis of the branched pentamannan core of the HIV1-gp120 complex. 28985062 0 32 theme Glycosides 87:96 arg1 Activation 51:60 the Remote Activation 40:60 the Remote Activation of Hydroxybenzotriazolyl Glycosides 40:96 Nucleofuge Generating Glycosidations by the Remote Activation of Hydroxybenzotriazolyl Glycosides. 28985062 0 33 theme Hydroxybenzotriazolyl 65:85 arg1 Glycosides 87:96 Hydroxybenzotriazolyl Glycosides 65:96 Hydroxybenzotriazolyl Glycosides 65:96 Nucleofuge Generating Glycosidations by the Remote Activation of Hydroxybenzotriazolyl Glycosides. 25883177 7 0 gly multifucosylated 1459:1474 arg1 antennae 1476:1483 multifucosylated antennae 1459:1483 multifucosylated antennae only in mature eggs and miracidia 1459:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 10 1 theme subshell 1975:1982 arg1 envelope 1984:1991 the subshell envelope 1971:1991 the subshell envelope in the developed egg 1971:2012 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 11 2 theme Lipid 2015:2019 arg1 glycans 2021:2027 Lipid glycans 2015:2027 Lipid glycans with multifucosylated GlcNAc repeats 2015:2064 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 6 3 theme O-glycans 1243:1251 arg1 expression 1221:1230 expression 1221:1230 expression of complex O-glycans 1221:1251 In contrast, expression of complex O-glycans diminished to undetectable levels within days after transformation. 25883177 12 4 theme expressed 2306:2314 arg1 glycans 2316:2322 stage-specifically expressed glycans 2287:2322 stage-specifically expressed glycans 2287:2322 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 1 5 attach present 168:174 arg2 Glycans 160:166 Glycans 160:166 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni 160:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 1 5 attach present 168:174 arg1 glycoproteins 179:191 glycoproteins 179:191 glycoproteins 179:191 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 1 5 attach present 168:174 arg1 glycolipids 197:207 glycolipids 197:207 glycolipids 197:207 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 11 6 theme egg 2090:2092 arg1 development 2094:2104 egg development 2090:2104 egg development 2090:2104 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 7 7 from miracidia 1509:1517 arg1 fucose 1431:1436 α3-core fucose 1423:1436 α3-core fucose 1423:1436 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 7 from miracidia 1509:1517 arg1 degree 1449:1454 a high degree 1442:1454 a high degree of multifucosylated antennae only in mature eggs and miracidia 1442:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 8 theme antennae 1476:1483 arg1 fucose 1431:1436 α3-core fucose 1423:1436 α3-core fucose 1423:1436 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 8 theme antennae 1476:1483 arg1 degree 1449:1454 a high degree 1442:1454 a high degree of multifucosylated antennae only in mature eggs and miracidia 1442:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 5 9 dep weeks 1064:1068 arg1 up 1056:1057 up 1056:1057 up 1056:1057 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 7 10 with diversity 1352:1360 arg1 motifs 1392:1397 fucosylated motifs 1380:1397 fucosylated motifs 1380:1397 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 3 11 theme longitudinal 585:596 arg1 study 608:612 a longitudinal profiling study 583:612 a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development 583:684 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 10 12 gly glycoproteins 1945:1957 arg1 glycoproteins 1945:1957 glycoproteins 1945:1957 glycoproteins produced by the subshell envelope in the developed egg 1945:2012 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 0 13 theme Functional 120:129 arg1 Motifs 152:157 Functional and Antigenic Glycan Motifs 120:157 Functional and Antigenic Glycan Motifs 120:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 5 14 theme mono- 1125:1129 arg1 LDN-motifs 1152:1161 mono- and multifucosylated LDN-motifs 1125:1161 mono- and multifucosylated LDN-motifs 1125:1161 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 10 15 theme O-glycan 1821:1828 arg1 structures 1835:1844 short O-glycan core structures 1815:1844 short O-glycan core structures 1815:1844 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 5 16 attach present 1033:1039 arg1 glycolipids 1044:1054 glycolipids 1044:1054 glycolipids up to 2 weeks of schistosomula development 1044:1097 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 5 16 attach present 1033:1039 arg2 LeX-motifs 1017:1026 LeX-motifs 1017:1026 LeX-motifs 1017:1026 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 2 17 theme schistosome 547:557 arg1 lifecycle 559:567 the complex schistosome lifecycle 535:567 the complex schistosome lifecycle 535:567 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 7 18 theme high 1444:1447 arg1 degree 1449:1454 a high degree 1442:1454 a high degree of multifucosylated antennae only in mature eggs and miracidia 1442:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 12 19 theme global 2200:2205 arg1 analysis 2207:2214 This global analysis 2195:2214 This global analysis of the developing schistosome's glycome 2195:2254 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 0 20 theme Antigenic 135:143 arg1 Motifs 152:157 Functional and Antigenic Glycan Motifs 120:157 Functional and Antigenic Glycan Motifs 120:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 5 21 theme multifucosylated 1135:1150 arg1 LDN-motifs 1152:1161 mono- and multifucosylated LDN-motifs 1125:1161 mono- and multifucosylated LDN-motifs 1125:1161 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 3 22 theme schistosome 623:633 arg1 glycosylation 635:647 schistosome glycosylation 623:647 schistosome glycosylation 623:647 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 1 23 theme human 222:226 arg1 mansoni 249:255 the major human parasite Schistosoma mansoni 212:255 the major human parasite Schistosoma mansoni 212:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 9 24 theme core 1773:1776 arg1 glycan 1778:1783 only the Galβ1-3(Galβ1-6)GalNAc core glycan 1741:1783 only the Galβ1-3(Galβ1-6)GalNAc core glycan 1741:1783 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 8 25 theme N-glycan 1520:1527 arg1 antennae 1529:1536 N-glycan antennae 1520:1536 N-glycan antennae 1520:1536 N-glycan antennae were exclusively LDN-based in miracidia. 25883177 4 26 theme worm 769:772 arg1 N-glycans 786:794 worm development N-glycans 769:794 worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs 769:851 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 0 27 theme Motifs 152:157 arg1 Profiles 108:115 Developmental Expression Profiles 83:115 Developmental Expression Profiles of Functional and Antigenic Glycan Motifs 83:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 7 28 theme α3-core 1423:1429 arg1 fucose 1431:1436 α3-core fucose 1423:1436 α3-core fucose 1423:1436 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 29 from antennae 1476:1483 arg1 miracidia 1509:1517 miracidia 1509:1517 miracidia 1509:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 29 from antennae 1476:1483 arg1 eggs 1500:1503 mature eggs 1493:1503 mature eggs 1493:1503 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 0 30 theme Parasite 46:53 arg1 mansoni 67:73 the Human Parasite Schistosoma mansoni 36:73 the Human Parasite Schistosoma mansoni 36:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 10 31 theme fucosylated 1876:1886 arg1 O-glycans 1888:1896 complex fucosylated O-glycans 1868:1896 complex fucosylated O-glycans of schistosome eggs 1868:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 9 32 theme mature 1596:1601 arg1 eggs 1603:1606 the mature eggs 1592:1606 the mature eggs 1592:1606 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 0 33 theme mansoni 67:73 arg1 Stages 26:31 Life Stages 21:31 Life Stages of the Human Parasite Schistosoma mansoni 21:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 5 34 theme worm 1196:1199 arg1 stage 1201:1205 the adult worm stage 1186:1205 the adult worm stage 1186:1205 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 1 35 theme present 168:174 arg1 Glycans 160:166 Glycans 160:166 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni 160:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 7 36 from eggs 1500:1503 arg1 fucose 1431:1436 α3-core fucose 1423:1436 α3-core fucose 1423:1436 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 36 from eggs 1500:1503 arg1 degree 1449:1454 a high degree 1442:1454 a high degree of multifucosylated antennae only in mature eggs and miracidia 1442:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 4 37 theme adult 1004:1008 arg1 worms 1010:1014 adult worms 1004:1014 adult worms 1004:1014 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 0 38 theme Developmental 83:95 arg1 Profiles 108:115 Developmental Expression Profiles 83:115 Developmental Expression Profiles of Functional and Antigenic Glycan Motifs 83:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 2 39 theme stages 525:530 arg1 range 507:511 a range 505:511 a range of critical stages of the complex schistosome lifecycle 505:567 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 12 40 theme interactions 2380:2391 arg1 aspects 2352:2358 different aspects 2342:2358 different aspects of schistosome-host interactions 2342:2391 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 4 41 theme Galβ1-4 801:807 arg1 GlcNAc 817:822 Galβ1-4(Fucα1-3)GlcNAc 801:822 Galβ1-4(Fucα1-3)GlcNAc (LeX) 801:828 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 4 41 theme Galβ1-4 801:807 arg1 LeX 825:827 LeX 825:827 LeX 825:827 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 2 42 theme infections 386:395 arg1 characteristics 355:369 the molecular characteristics 341:369 the molecular characteristics of schistosome infections 341:395 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 0 43 theme Glycomic 0:7 arg1 Analysis 9:16 Glycomic Analysis 0:16 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni 0:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 1 44 theme adaptive 282:289 arg1 responses 298:306 innate as well as adaptive immune responses 264:306 innate as well as adaptive immune responses 264:306 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 2 45 theme expression 439:448 arg1 profiles 450:457 the expression profiles 435:457 the expression profiles of glycans and antigenic glycan-motifs 435:496 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 5 46 dep stage 1201:1205 arg1 up 1180:1181 up 1180:1181 up 1180:1181 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 4 47 theme Fucα1-3 809:815 arg1 GlcNAc 817:822 Galβ1-4(Fucα1-3)GlcNAc 801:822 Galβ1-4(Fucα1-3)GlcNAc (LeX) 801:828 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 4 47 theme Fucα1-3 809:815 arg1 LeX 825:827 LeX 825:827 LeX 825:827 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 7 48 theme fucosylated 1380:1390 arg1 motifs 1392:1397 fucosylated motifs 1380:1397 fucosylated motifs 1380:1397 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 2 49 theme glycan-motifs 484:496 arg1 profiles 450:457 the expression profiles 435:457 the expression profiles of glycans and antigenic glycan-motifs 435:496 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 3 50 theme mass 694:697 arg1 approach 728:735 a mass spectrometry-based glycomics approach 692:735 a mass spectrometry-based glycomics approach 692:735 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 2 51 dep required 413:420 arg1 able 327:330 able 327:330 able 327:330 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 0 52 theme Stages 26:31 arg1 Analysis 9:16 Glycomic Analysis 0:16 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni 0:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 7 53 theme N-glycans 1365:1373 arg1 expressed 1403:1411 expressed 1403:1411 expressed 1403:1411 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 53 theme N-glycans 1365:1373 arg1 diversity 1352:1360 a rich diversity 1345:1360 a rich diversity of N-glycans with fucosylated motifs 1345:1397 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 10 54 theme eggs 1913:1916 arg1 O-glycans 1888:1896 complex fucosylated O-glycans 1868:1896 complex fucosylated O-glycans of schistosome eggs 1868:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 10 55 theme developed 2000:2008 arg1 egg 2010:2012 the developed egg 1996:2012 the developed egg 1996:2012 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 5 56 theme development 1087:1097 arg1 weeks 1064:1068 2 weeks 1062:1068 2 weeks of schistosomula development 1062:1097 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 9 57 gly multifucosylated 1649:1664 arg1 LDN 1666:1668 multifucosylated LDN 1649:1668 multifucosylated LDN 1649:1668 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 12 58 theme new 2265:2267 arg1 insights 2269:2276 new insights 2265:2276 new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions 2265:2391 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 1 59 theme parasite 228:235 arg1 mansoni 249:255 the major human parasite Schistosoma mansoni 212:255 the major human parasite Schistosoma mansoni 212:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 9 60 contain contained 1630:1638 arg1 O-glycans 1579:1587 O-glycans 1579:1587 O-glycans in the mature eggs 1579:1606 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 9 60 contain contained 1630:1638 arg2 LDN 1666:1668 multifucosylated LDN 1649:1668 multifucosylated LDN 1649:1668 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 9 60 contain contained 1630:1638 arg2 LeX- 1640:1643 LeX- 1640:1643 LeX- 1640:1643 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 3 61 theme glycomics 718:726 arg1 approach 728:735 a mass spectrometry-based glycomics approach 692:735 a mass spectrometry-based glycomics approach 692:735 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 10 62 theme Immature 1786:1793 arg1 eggs 1795:1798 Immature eggs 1786:1798 Immature eggs 1786:1798 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 1 63 theme mansoni 249:255 arg1 glycolipids 197:207 glycolipids 197:207 glycolipids 197:207 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 1 63 theme mansoni 249:255 arg1 glycoproteins 179:191 glycoproteins 179:191 glycoproteins 179:191 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 1 64 gly glycoproteins 179:191 arg1 glycoproteins 179:191 glycoproteins 179:191 glycoproteins 179:191 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 2 65 theme molecular 345:353 arg1 characteristics 355:369 the molecular characteristics 341:369 the molecular characteristics of schistosome infections 341:395 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 1 66 theme innate 264:269 arg1 responses 298:306 innate as well as adaptive immune responses 264:306 innate as well as adaptive immune responses 264:306 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 10 67 theme short 1815:1819 arg1 structures 1835:1844 short O-glycan core structures 1815:1844 short O-glycan core structures 1815:1844 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 4 68 theme GalNAcβ1-4GlcNAc 928:943 arg1 -motifs 950:956 GalNAcβ1-4GlcNAc (LDN)-motifs 928:956 GalNAcβ1-4GlcNAc (LDN)-motifs 928:956 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 6 69 theme complex 1235:1241 arg1 O-glycans 1243:1251 complex O-glycans 1235:1251 complex O-glycans 1235:1251 In contrast, expression of complex O-glycans diminished to undetectable levels within days after transformation. 25883177 12 70 theme developing 2223:2232 arg1 glycome 2248:2254 the developing schistosome's glycome 2219:2254 the developing schistosome's glycome 2219:2254 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 9 71 from O-glycans 1579:1587 arg1 eggs 1603:1606 the mature eggs 1592:1606 the mature eggs 1592:1606 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 7 72 theme mature 1493:1498 arg1 eggs 1500:1503 mature eggs 1493:1503 mature eggs 1493:1503 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 11 73 with present 2071:2077 arg1 stretches 2146:2154 the longer highly fucosylated stretches 2116:2154 the longer highly fucosylated stretches enriched in mature eggs and miracidia 2116:2192 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 4 74 theme schistosomula 890:902 arg1 transformation 904:917 schistosomula transformation 890:917 schistosomula transformation 890:917 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 10 75 theme core 1830:1833 arg1 structures 1835:1844 short O-glycan core structures 1815:1844 short O-glycan core structures 1815:1844 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 10 76 from envelope 1984:1991 arg1 egg 2010:2012 the developed egg 1996:2012 the developed egg 1996:2012 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 2 77 theme lifecycle 559:567 arg1 stages 525:530 critical stages 516:530 critical stages of the complex schistosome lifecycle 516:567 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 11 78 theme fucosylated 2134:2144 arg1 stretches 2146:2154 the longer highly fucosylated stretches 2116:2154 the longer highly fucosylated stretches enriched in mature eggs and miracidia 2116:2192 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 7 79 theme multifucosylated 1459:1474 arg1 antennae 1476:1483 multifucosylated antennae 1459:1483 multifucosylated antennae only in mature eggs and miracidia 1459:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 0 80 theme Glycan 145:150 arg1 Motifs 152:157 Functional and Antigenic Glycan Motifs 120:157 Functional and Antigenic Glycan Motifs 120:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 11 81 gly fucosylated 2134:2144 arg1 stretches 2146:2154 the longer highly fucosylated stretches 2116:2154 the longer highly fucosylated stretches enriched in mature eggs and miracidia 2116:2192 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 10 82 theme complex 1868:1874 arg1 O-glycans 1888:1896 complex fucosylated O-glycans 1868:1896 complex fucosylated O-glycans of schistosome eggs 1868:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 1 83 theme major 216:220 arg1 mansoni 249:255 the major human parasite Schistosoma mansoni 212:255 the major human parasite Schistosoma mansoni 212:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 2 84 theme complex 539:545 arg1 lifecycle 559:567 the complex schistosome lifecycle 535:567 the complex schistosome lifecycle 535:567 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 3 85 theme profiling 598:606 arg1 study 608:612 a longitudinal profiling study 583:612 a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development 583:684 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 4 86 theme LDN 946:948 arg1 -motifs 950:956 GalNAcβ1-4GlcNAc (LDN)-motifs 928:956 GalNAcβ1-4GlcNAc (LDN)-motifs 928:956 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 10 87 theme schistosome 1901:1911 arg1 eggs 1913:1916 schistosome eggs 1901:1916 schistosome eggs 1901:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 11 88 theme mature 2168:2173 arg1 eggs 2175:2178 mature eggs 2168:2178 mature eggs 2168:2178 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 6 89 theme undetectable 1267:1278 arg1 levels 1280:1285 undetectable levels 1267:1285 undetectable levels 1267:1285 In contrast, expression of complex O-glycans diminished to undetectable levels within days after transformation. 25883177 4 90 theme development 774:784 arg1 N-glycans 786:794 worm development N-glycans 769:794 worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs 769:851 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 0 91 theme Schistosoma 55:65 arg1 mansoni 67:73 the Human Parasite Schistosoma mansoni 36:73 the Human Parasite Schistosoma mansoni 36:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 11 92 theme GlcNAc 2051:2056 arg1 repeats 2058:2064 multifucosylated GlcNAc repeats 2034:2064 multifucosylated GlcNAc repeats 2034:2064 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 10 93 attach derived 1922:1928 arg1 glycoproteins 1945:1957 glycoproteins 1945:1957 glycoproteins produced by the subshell envelope in the developed egg 1945:2012 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 10 93 attach derived 1922:1928 arg2 O-glycans 1888:1896 complex fucosylated O-glycans 1868:1896 complex fucosylated O-glycans of schistosome eggs 1868:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 11 94 with glycans 2021:2027 arg1 repeats 2058:2064 multifucosylated GlcNAc repeats 2034:2064 multifucosylated GlcNAc repeats 2034:2064 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 5 95 theme adult 1190:1194 arg1 stage 1201:1205 the adult worm stage 1186:1205 the adult worm stage 1186:1205 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 7 96 gly fucosylated 1380:1390 arg1 motifs 1392:1397 fucosylated motifs 1380:1397 fucosylated motifs 1380:1397 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 0 97 theme Expression 97:106 arg1 Profiles 108:115 Developmental Expression Profiles 83:115 Developmental Expression Profiles of Functional and Antigenic Glycan Motifs 83:157 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 12 98 theme schistosome-host 2363:2378 arg1 interactions 2380:2391 schistosome-host interactions 2363:2391 schistosome-host interactions 2363:2391 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 5 99 with glycolipids 1108:1118 arg1 LDN-motifs 1152:1161 mono- and multifucosylated LDN-motifs 1125:1161 mono- and multifucosylated LDN-motifs 1125:1161 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 2 100 theme critical 516:523 arg1 stages 525:530 critical stages 516:530 critical stages of the complex schistosome lifecycle 516:567 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 12 101 theme different 2342:2350 arg1 aspects 2352:2358 different aspects 2342:2358 different aspects of schistosome-host interactions 2342:2391 This global analysis of the developing schistosome's glycome provides new insights into how stage-specifically expressed glycans may contribute to different aspects of schistosome-host interactions. 25883177 7 102 from fucose 1431:1436 arg1 miracidia 1509:1517 miracidia 1509:1517 miracidia 1509:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 102 from fucose 1431:1436 arg1 eggs 1500:1503 mature eggs 1493:1503 mature eggs 1493:1503 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 9 103 theme GalNAc 1766:1771 arg1 glycan 1778:1783 only the Galβ1-3(Galβ1-6)GalNAc core glycan 1741:1783 only the Galβ1-3(Galβ1-6)GalNAc core glycan 1741:1783 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 2 104 theme schistosome 374:384 arg1 infections 386:395 schistosome infections 374:395 schistosome infections 374:395 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 0 105 theme Life 21:24 arg1 Stages 26:31 Life Stages 21:31 Life Stages of the Human Parasite Schistosoma mansoni 21:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 1 106 theme immune 291:296 arg1 responses 298:306 innate as well as adaptive immune responses 264:306 innate as well as adaptive immune responses 264:306 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 9 107 theme multifucosylated 1649:1664 arg1 LDN 1666:1668 multifucosylated LDN 1649:1668 multifucosylated LDN 1649:1668 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 5 108 located present 1033:1039 arg1 glycolipids 1044:1054 glycolipids 1044:1054 glycolipids up to 2 weeks of schistosomula development 1044:1097 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 5 108 located present 1033:1039 arg2 LeX-motifs 1017:1026 LeX-motifs 1017:1026 LeX-motifs 1017:1026 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 7 109 from degree 1449:1454 arg1 miracidia 1509:1517 miracidia 1509:1517 miracidia 1509:1517 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 109 from degree 1449:1454 arg1 eggs 1500:1503 mature eggs 1493:1503 mature eggs 1493:1503 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 2 110 theme antigenic 474:482 arg1 glycan-motifs 484:496 antigenic glycan-motifs 474:496 antigenic glycan-motifs 474:496 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 11 111 gly multifucosylated 2034:2049 arg1 repeats 2058:2064 multifucosylated GlcNAc repeats 2034:2064 multifucosylated GlcNAc repeats 2034:2064 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 10 112 gly fucosylated 1876:1886 arg1 O-glycans 1888:1896 complex fucosylated O-glycans 1868:1896 complex fucosylated O-glycans of schistosome eggs 1868:1916 Immature eggs also exhibited short O-glycan core structures only, suggesting that complex fucosylated O-glycans of schistosome eggs are derived primarily from glycoproteins produced by the subshell envelope in the developed egg. 25883177 0 113 theme Human 40:44 arg1 mansoni 67:73 the Human Parasite Schistosoma mansoni 36:73 the Human Parasite Schistosoma mansoni 36:73 Glycomic Analysis of Life Stages of the Human Parasite Schistosoma mansoni Reveals Developmental Expression Profiles of Functional and Antigenic Glycan Motifs. 25883177 5 114 theme schistosomula 1073:1085 arg1 development 1087:1097 schistosomula development 1073:1097 schistosomula development 1073:1097 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 7 115 theme rich 1347:1350 arg1 expressed 1403:1411 expressed 1403:1411 expressed 1403:1411 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 7 115 theme rich 1347:1350 arg1 diversity 1352:1360 a rich diversity 1345:1360 a rich diversity of N-glycans with fucosylated motifs 1345:1397 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 2 116 theme glycans 462:468 arg1 profiles 450:457 the expression profiles 435:457 the expression profiles of glycans and antigenic glycan-motifs 435:496 To be able to study the molecular characteristics of schistosome infections it is therefore required to determine the expression profiles of glycans and antigenic glycan-motifs during a range of critical stages of the complex schistosome lifecycle. 25883177 9 117 theme these 1683:1687 arg1 none 1675:1678 none 1675:1678 none of these 1675:1687 O-glycans in the mature eggs were also diverse and contained LeX- and multifucosylated LDN, but none of these were associated with miracidia in which we detected only the Galβ1-3(Galβ1-6)GalNAc core glycan. 25883177 11 118 theme multifucosylated 2034:2049 arg1 repeats 2058:2064 multifucosylated GlcNAc repeats 2034:2064 multifucosylated GlcNAc repeats 2034:2064 Lipid glycans with multifucosylated GlcNAc repeats were present throughout egg development, but with the longer highly fucosylated stretches enriched in mature eggs and miracidia. 25883177 3 119 theme spectrometry-based 699:716 arg1 approach 728:735 a mass spectrometry-based glycomics approach 692:735 a mass spectrometry-based glycomics approach 692:735 We performed a longitudinal profiling study covering schistosome glycosylation throughout worm- and egg-development using a mass spectrometry-based glycomics approach. 25883177 1 120 theme Schistosoma 237:247 arg1 mansoni 249:255 the major human parasite Schistosoma mansoni 212:255 the major human parasite Schistosoma mansoni 212:255 Glycans present on glycoproteins and glycolipids of the major human parasite Schistosoma mansoni induce innate as well as adaptive immune responses in the host. 25883177 7 121 theme egg 1328:1330 arg1 development 1332:1342 egg development 1328:1342 egg development 1328:1342 During egg development, a rich diversity of N-glycans with fucosylated motifs was expressed, but with α3-core fucose and a high degree of multifucosylated antennae only in mature eggs and miracidia. 25883177 5 122 gly multifucosylated 1135:1150 arg1 LDN-motifs 1152:1161 mono- and multifucosylated LDN-motifs 1125:1161 mono- and multifucosylated LDN-motifs 1125:1161 LeX-motifs were present on glycolipids up to 2 weeks of schistosomula development, whereas glycolipids with mono- and multifucosylated LDN-motifs remained present up to the adult worm stage. 25883177 4 123 with N-glycans 786:794 arg1 GlcNAc 817:822 Galβ1-4(Fucα1-3)GlcNAc 801:822 Galβ1-4(Fucα1-3)GlcNAc (LeX) 801:828 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 4 123 with N-glycans 786:794 arg1 motifs 846:851 core-xylose motifs 834:851 core-xylose motifs 834:851 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 4 123 with N-glycans 786:794 arg1 LeX 825:827 LeX 825:827 LeX 825:827 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 25883177 4 124 theme core-xylose 834:844 arg1 motifs 846:851 core-xylose motifs 834:851 core-xylose motifs 834:851 Our study revealed that during worm development N-glycans with Galβ1-4(Fucα1-3)GlcNAc (LeX) and core-xylose motifs were rapidly lost after cercariae to schistosomula transformation, whereas GalNAcβ1-4GlcNAc (LDN)-motifs gradually became abundant and predominated in adult worms. 27305319 5 0 theme consistent 717:726 arg1 use 728:730 consistent use 717:730 consistent use of oligosaccharyl thioether and oligosaccharyl bromide 717:785 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 1 theme versatile 640:648 arg1 blocks 659:664 3 versatile building blocks 638:664 3 versatile building blocks 638:664 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 1 2 theme glycans 142:148 arg1 family 111:116 a family 109:116 a family of diverse unconjugated glycans that exist in human milk as one of the major components 109:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 2 theme glycans 142:148 arg1 oligosaccharides 81:96 Human milk oligosaccharides 70:96 Human milk oligosaccharides (HMOs) 70:103 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 5 3 dep blocks 659:664 arg1 basis 629:633 basis 629:633 basis 629:633 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 3 dep blocks 659:664 arg1 the 625:627 the 625:627 the 625:627 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 4 4 theme extension 553:561 arg1 CSEE 564:567 namely core synthesis/enzymatic extension (CSEE) 521:568 namely core synthesis/enzymatic extension (CSEE) 521:568 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 4 4 theme extension 553:561 arg1 strategy 511:518 an efficient chemoenzymatic strategy 483:518 an efficient chemoenzymatic strategy 483:518 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 5 5 theme core 669:672 arg1 donors 804:809 glycosylation donors 790:809 glycosylation donors 790:809 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 5 theme core 669:672 arg1 structures 674:683 3 core structures 667:683 3 core structures 667:683 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 3 6 theme homogeneous 362:372 arg1 library 378:384 a homogeneous HMO library 360:384 a homogeneous HMO library 360:384 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 2 7 theme HMOs 270:273 arg1 quantification 225:238 quantification 225:238 quantification 225:238 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 2 7 theme HMOs 270:273 arg1 Characterization 207:222 Characterization 207:222 Characterization 207:222 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 2 7 theme HMOs 270:273 arg1 studies 259:265 biofunctional studies 245:265 biofunctional studies of HMOs 245:273 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 6 8 theme core 869:872 arg1 structures 874:883 these core structures 863:883 these core structures 863:883 Each of these core structures was then extended to up to 11 HMOs by 4 robust glycosyltransferases. 27305319 3 9 theme several 451:457 arg1 decades 459:465 several decades 451:465 several decades 451:465 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 1 10 theme Human 70:74 arg1 family 111:116 a family 109:116 a family of diverse unconjugated glycans that exist in human milk as one of the major components 109:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 10 theme Human 70:74 arg1 HMOs 99:102 HMOs 99:102 HMOs 99:102 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 10 theme Human 70:74 arg1 oligosaccharides 81:96 Human milk oligosaccharides 70:96 Human milk oligosaccharides (HMOs) 70:103 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 5 11 theme fragment 827:834 arg1 strategy 845:852 a convergent fragment coupling strategy 814:852 a convergent fragment coupling strategy 814:852 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 0 12 theme Human 41:45 arg1 Oligosaccharides 52:67 Human Milk Oligosaccharides 41:67 Human Milk Oligosaccharides 41:67 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides. 27305319 1 13 theme milk 76:79 arg1 family 111:116 a family 109:116 a family of diverse unconjugated glycans that exist in human milk as one of the major components 109:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 13 theme milk 76:79 arg1 HMOs 99:102 HMOs 99:102 HMOs 99:102 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 13 theme milk 76:79 arg1 oligosaccharides 81:96 Human milk oligosaccharides 70:96 Human milk oligosaccharides (HMOs) 70:103 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 5 14 theme coupling 836:843 arg1 strategy 845:852 a convergent fragment coupling strategy 814:852 a convergent fragment coupling strategy 814:852 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 15 theme oligosaccharyl 764:777 arg1 bromide 779:785 oligosaccharyl bromide 764:785 oligosaccharyl bromide 764:785 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 8 16 theme defined 1110:1116 arg1 HMOs 1118:1121 structurally defined HMOs 1097:1121 structurally defined HMOs 1097:1121 CSEE indeed provides a practical approach to harvest structurally defined HMOs for various applications. 27305319 1 17 theme human 164:168 arg1 milk 170:173 human milk 164:173 human milk 164:173 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 0 18 theme Chemoenzymatic 0:13 arg1 Synthesis 15:23 Chemoenzymatic Synthesis 0:23 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides 0:67 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides. 27305319 6 19 theme robust 925:930 arg1 glycosyltransferases 932:951 4 robust glycosyltransferases 923:951 4 robust glycosyltransferases 923:951 Each of these core structures was then extended to up to 11 HMOs by 4 robust glycosyltransferases. 27305319 8 20 dep harvest 1089:1095 arg1 HMOs 1118:1121 structurally defined HMOs 1097:1121 structurally defined HMOs 1097:1121 CSEE indeed provides a practical approach to harvest structurally defined HMOs for various applications. 27305319 2 21 theme great 284:288 arg1 challenge 290:298 a great challenge 282:298 a great challenge due to their diversity and complexity 282:336 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 5 22 theme bromide 779:785 arg1 use 728:730 consistent use 717:730 consistent use of oligosaccharyl thioether and oligosaccharyl bromide 717:785 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 23 theme building 650:657 arg1 blocks 659:664 3 versatile building blocks 638:664 3 versatile building blocks 638:664 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 24 theme thioether 750:758 arg1 use 728:730 consistent use 717:730 consistent use of oligosaccharyl thioether and oligosaccharyl bromide 717:785 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 3 25 theme beset 432:436 arg1 academia 438:445 beset academia 432:445 beset academia 432:445 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 5 26 theme glycosylation 790:802 arg1 donors 804:809 glycosylation donors 790:809 glycosylation donors 790:809 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 5 26 theme glycosylation 790:802 arg1 structures 674:683 3 core structures 667:683 3 core structures 667:683 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 4 27 theme synthesis/enzymatic 533:551 arg1 CSEE 564:567 namely core synthesis/enzymatic extension (CSEE) 521:568 namely core synthesis/enzymatic extension (CSEE) 521:568 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 4 27 theme synthesis/enzymatic 533:551 arg1 strategy 511:518 an efficient chemoenzymatic strategy 483:518 an efficient chemoenzymatic strategy 483:518 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 3 28 contain have 427:430 arg2 academia 438:445 beset academia 432:445 beset academia 432:445 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 3 28 contain have 427:430 arg1 issues 414:419 these issues 408:419 these issues which have beset academia for several decades 408:465 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 4 29 theme core 528:531 arg1 CSEE 564:567 namely core synthesis/enzymatic extension (CSEE) 521:568 namely core synthesis/enzymatic extension (CSEE) 521:568 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 4 29 theme core 528:531 arg1 strategy 511:518 an efficient chemoenzymatic strategy 483:518 an efficient chemoenzymatic strategy 483:518 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 0 30 theme Library 30:36 arg1 Synthesis 15:23 Chemoenzymatic Synthesis 0:23 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides 0:67 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides. 27305319 4 31 theme efficient 486:494 arg1 CSEE 564:567 namely core synthesis/enzymatic extension (CSEE) 521:568 namely core synthesis/enzymatic extension (CSEE) 521:568 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 4 31 theme efficient 486:494 arg1 strategy 511:518 an efficient chemoenzymatic strategy 483:518 an efficient chemoenzymatic strategy 483:518 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 5 32 theme oligosaccharyl 735:748 arg1 thioether 750:758 oligosaccharyl thioether 735:758 oligosaccharyl thioether 735:758 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 6 33 dep 11 912:913 arg1 to 909:910 to 909:910 to 909:910 Each of these core structures was then extended to up to 11 HMOs by 4 robust glycosyltransferases. 27305319 7 34 theme HMOs 970:973 arg1 library 956:962 A library 954:962 A library of 31 HMOs 954:973 A library of 31 HMOs were chemoenzymatically synthesized and characterized by MS and NMR. 27305319 4 35 theme HMOs 603:606 arg1 production 581:590 rapid production 575:590 rapid production of diverse HMOs 575:606 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 3 36 theme HMO 374:376 arg1 library 378:384 a homogeneous HMO library 360:384 a homogeneous HMO library 360:384 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 8 37 theme various 1127:1133 arg1 applications 1135:1146 various applications 1127:1146 various applications 1127:1146 CSEE indeed provides a practical approach to harvest structurally defined HMOs for various applications. 27305319 8 38 theme practical 1067:1075 arg1 approach 1077:1084 a practical approach 1065:1084 a practical approach to harvest structurally defined HMOs for various applications 1065:1146 CSEE indeed provides a practical approach to harvest structurally defined HMOs for various applications. 27305319 0 39 theme Oligosaccharides 52:67 arg1 Library 30:36 a Library 28:36 a Library of Human Milk Oligosaccharides 28:67 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides. 27305319 3 40 theme library 378:384 arg1 accessibility 343:355 The accessibility 339:355 The accessibility of a homogeneous HMO library 339:384 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 3 40 theme library 378:384 arg1 essential 389:397 essential 389:397 essential 389:397 The accessibility of a homogeneous HMO library is essential to solve these issues which have beset academia for several decades. 27305319 4 41 theme chemoenzymatic 496:509 arg1 CSEE 564:567 namely core synthesis/enzymatic extension (CSEE) 521:568 namely core synthesis/enzymatic extension (CSEE) 521:568 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 4 41 theme chemoenzymatic 496:509 arg1 strategy 511:518 an efficient chemoenzymatic strategy 483:518 an efficient chemoenzymatic strategy 483:518 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 1 42 theme major 189:193 arg1 components 195:204 the major components 185:204 the major components 185:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 2 43 theme biofunctional 245:257 arg1 studies 259:265 biofunctional studies 245:265 biofunctional studies of HMOs 245:273 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 2 44 theme due 300:302 arg1 challenge 290:298 a great challenge 282:298 a great challenge due to their diversity and complexity 282:336 Characterization, quantification, and biofunctional studies of HMOs remain a great challenge due to their diversity and complexity. 27305319 1 45 theme components 195:204 arg1 one 178:180 one 178:180 one 178:180 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 1 45 theme components 195:204 arg1 components 195:204 the major components 185:204 the major components 185:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 5 46 theme convergent 816:825 arg1 strategy 845:852 a convergent fragment coupling strategy 814:852 a convergent fragment coupling strategy 814:852 On the basis of 3 versatile building blocks, 3 core structures were chemically synthesized via consistent use of oligosaccharyl thioether and oligosaccharyl bromide as glycosylation donors in a convergent fragment coupling strategy. 27305319 4 47 theme diverse 595:601 arg1 HMOs 603:606 diverse HMOs 595:606 diverse HMOs 595:606 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 27305319 1 48 theme diverse 121:127 arg1 glycans 142:148 diverse unconjugated glycans 121:148 diverse unconjugated glycans that exist in human milk as one of the major components 121:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 0 49 theme Milk 47:50 arg1 Oligosaccharides 52:67 Human Milk Oligosaccharides 41:67 Human Milk Oligosaccharides 41:67 Chemoenzymatic Synthesis of a Library of Human Milk Oligosaccharides. 27305319 1 50 theme unconjugated 129:140 arg1 glycans 142:148 diverse unconjugated glycans 121:148 diverse unconjugated glycans that exist in human milk as one of the major components 121:204 Human milk oligosaccharides (HMOs) are a family of diverse unconjugated glycans that exist in human milk as one of the major components. 27305319 4 51 theme rapid 575:579 arg1 production 581:590 rapid production 575:590 rapid production of diverse HMOs 575:606 In this study, an efficient chemoenzymatic strategy, namely core synthesis/enzymatic extension (CSEE), for rapid production of diverse HMOs was reported. 26998654 0 0 theme Key 91:93 arg1 Residues 95:102 Key Residues 91:102 Key Residues for Transglycosylating Activity 91:134 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 2 1 theme GOS 540:542 arg1 biosynthesis 544:555 GOS biosynthesis 540:555 GOS biosynthesis 540:555 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 3 2 theme Rational 558:565 arg1 design 567:572 Rational design 558:572 Rational design of mutations 558:585 Rational design of mutations was based on homologous sequence analysis and structural modeling. 26998654 4 3 from position 753:760 arg1 critical 766:773 critical 766:773 critical 766:773 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 3 from position 753:760 arg1 residue 696:702 residue W959 696:707 residue W959 696:707 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 3 from position 753:760 arg1 residue 737:743 an alternative aromatic residue 713:743 an alternative aromatic residue at this position 713:760 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 5 4 theme reduced 925:931 arg1 synthesis 933:941 an 80% reduced synthesis 918:941 an 80% reduced synthesis of this GOS 918:953 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 7 5 theme 2-fold 1135:1140 arg1 increase 1117:1124 an increase 1114:1124 an increase of about 2-fold 1114:1140 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 1 6 theme secondary 278:286 arg1 activity 289:296 the transglycosylating (secondary) activity 254:296 the transglycosylating (secondary) activity of these enzymes 254:313 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 4 7 theme major 825:829 arg1 β-3'-galactosyl-lactose 796:818 β-3'-galactosyl-lactose 796:818 β-3'-galactosyl-lactose 796:818 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 7 theme major 825:829 arg1 GOS 831:833 the major GOS 821:833 the major GOS obtained with the wild-type enzyme 821:868 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 8 theme β-3'-galactosyl-lactose 796:818 arg1 synthesis 783:791 the synthesis 779:791 the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme 779:868 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 5 9 dep Mutants 871:877 arg1 Mutants 871:877 Mutants 871:877 Mutants W959A and W959C 871:893 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 5 9 dep Mutants 871:877 arg1 W959C 889:893 W959C 889:893 W959C 889:893 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 5 9 dep Mutants 871:877 arg1 W959A 879:883 W959A 879:883 W959A 879:883 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 1 10 gly transglycosylating 258:275 arg0 activity 289:296 the transglycosylating (secondary) activity 254:296 the transglycosylating (secondary) activity of these enzymes 254:313 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 3 11 theme homologous 600:609 arg1 analysis 620:627 homologous sequence analysis 600:627 homologous sequence analysis 600:627 Rational design of mutations was based on homologous sequence analysis and structural modeling. 26998654 5 12 theme GOS 951:953 arg1 synthesis 933:941 an 80% reduced synthesis 918:941 an 80% reduced synthesis of this GOS 918:953 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 4 13 theme alternative 716:726 arg1 critical 766:773 critical 766:773 critical 766:773 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 13 theme alternative 716:726 arg1 residue 737:743 an alternative aromatic residue 713:743 an alternative aromatic residue at this position 713:760 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 0 14 theme Transglycosylating 108:125 arg1 Activity 127:134 Transglycosylating Activity 108:134 Transglycosylating Activity 108:134 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 6 15 theme β-3'-galactosyl-lactose 1029:1051 arg1 synthesis 1016:1024 the synthesis 1012:1024 the synthesis of β-3'-galactosyl-lactose 1012:1051 Other substitutions, N574S, N574A, and F571L, increased the synthesis of β-3'-galactosyl-lactose about 40%. 26998654 2 16 theme enzymatic 493:501 arg1 variants 503:510 enzymatic variants 493:510 enzymatic variants with better performance for GOS biosynthesis 493:555 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 3 17 theme mutations 577:585 arg1 design 567:572 Rational design 558:572 Rational design of mutations 558:585 Rational design of mutations was based on homologous sequence analysis and structural modeling. 26998654 2 18 from maritima 395:402 arg1 mutagenesis 330:340 Site-directed mutagenesis 316:340 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima 316:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 2 18 from maritima 395:402 arg1 β-galactosidase 363:377 a thermoresistant β-galactosidase 345:377 a thermoresistant β-galactosidase from Thermotoga maritima 345:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 5 19 dep reduced 925:931 arg1 % 923:923 80% 921:923 80% 921:923 Mutants W959A and W959C, but not W959F, showed an 80% reduced synthesis of this GOS. 26998654 0 20 theme Structural 0:9 arg1 Dissection 11:20 Structural Dissection 0:20 Structural Dissection of the Active Site of Thermotoga maritima 0:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 3 21 theme structural 633:642 arg1 modeling 644:651 structural modeling 633:651 structural modeling 633:651 Rational design of mutations was based on homologous sequence analysis and structural modeling. 26998654 4 22 theme aromatic 728:735 arg1 critical 766:773 critical 766:773 critical 766:773 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 4 22 theme aromatic 728:735 arg1 residue 737:743 an alternative aromatic residue 713:743 an alternative aromatic residue at this position 713:760 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 0 23 gly Transglycosylating 108:125 arg0 Activity 127:134 Transglycosylating Activity 108:134 Transglycosylating Activity 108:134 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 2 24 theme transgalactosylation 458:477 arg1 basis 449:453 the structural basis 434:453 the structural basis of transgalactosylation 434:477 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 1 25 theme enzymes 307:313 arg1 activity 289:296 the transglycosylating (secondary) activity 254:296 the transglycosylating (secondary) activity of these enzymes 254:313 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 4 26 theme enzymes 673:679 arg1 Analysis 654:661 Analysis 654:661 Analysis of mutant enzymes 654:679 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 2 27 theme Site-directed 316:328 arg1 mutagenesis 330:340 Site-directed mutagenesis 316:340 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima 316:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 0 28 theme Site 36:39 arg1 Dissection 11:20 Structural Dissection 0:20 Structural Dissection of the Active Site of Thermotoga maritima 0:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 0 29 theme Active 29:34 arg1 Site 36:39 the Active Site 25:39 the Active Site of Thermotoga maritima 25:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 0 29 theme Active 29:34 arg1 maritima 55:62 Thermotoga maritima 44:62 Thermotoga maritima 44:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 2 30 theme better 517:522 arg1 performance 524:534 better performance 517:534 better performance for GOS biosynthesis 517:555 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 2 31 theme structural 438:447 arg1 basis 449:453 the structural basis 434:453 the structural basis of transgalactosylation 434:477 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 0 32 theme Thermotoga 44:53 arg1 maritima 55:62 Thermotoga maritima 44:62 Thermotoga maritima 44:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 2 33 theme Thermotoga 384:393 arg1 maritima 395:402 Thermotoga maritima 384:402 Thermotoga maritima 384:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 7 34 theme Double 1064:1069 arg1 F571L/N574S 1079:1089 F571L/N574S 1079:1089 F571L/N574S 1079:1089 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 7 34 theme Double 1064:1069 arg1 mutants 1071:1077 Double mutants 1064:1077 Double mutants F571L/N574S and F571L/N574A 1064:1105 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 7 34 theme Double 1064:1069 arg1 F571L/N574A 1095:1105 F571L/N574A 1095:1105 F571L/N574A 1095:1105 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 1 35 theme Glycoside 137:145 arg1 hydrolases 147:156 Glycoside hydrolases 137:156 Glycoside hydrolases 137:156 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 6 36 theme Other 956:960 arg1 N574A 984:988 N574A 984:988 N574A 984:988 Other substitutions, N574S, N574A, and F571L, increased the synthesis of β-3'-galactosyl-lactose about 40%. 26998654 6 36 theme Other 956:960 arg1 F571L 995:999 F571L 995:999 F571L 995:999 Other substitutions, N574S, N574A, and F571L, increased the synthesis of β-3'-galactosyl-lactose about 40%. 26998654 6 36 theme Other 956:960 arg1 N574S 977:981 N574S 977:981 N574S 977:981 Other substitutions, N574S, N574A, and F571L, increased the synthesis of β-3'-galactosyl-lactose about 40%. 26998654 6 36 theme Other 956:960 arg1 substitutions 962:974 Other substitutions 956:974 Other substitutions 956:974 Other substitutions, N574S, N574A, and F571L, increased the synthesis of β-3'-galactosyl-lactose about 40%. 26998654 2 37 theme β-galactosidase 363:377 arg1 mutagenesis 330:340 Site-directed mutagenesis 316:340 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima 316:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 3 38 theme sequence 611:618 arg1 analysis 620:627 homologous sequence analysis 600:627 homologous sequence analysis 600:627 Rational design of mutations was based on homologous sequence analysis and structural modeling. 26998654 1 39 theme due 247:249 arg1 GOS 242:244 GOS 242:244 GOS 242:244 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 1 39 theme due 247:249 arg1 galacto-oligosaccharides 216:239 galacto-oligosaccharides 216:239 galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes 216:313 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 2 40 theme thermoresistant 347:361 arg1 β-galactosidase 363:377 a thermoresistant β-galactosidase 345:377 a thermoresistant β-galactosidase from Thermotoga maritima 345:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 4 41 theme mutant 666:671 arg1 enzymes 673:679 mutant enzymes 666:679 mutant enzymes 666:679 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 7 42 dep mutants 1071:1077 arg1 F571L/N574S 1079:1089 F571L/N574S 1079:1089 F571L/N574S 1079:1089 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 7 42 dep mutants 1071:1077 arg1 mutants 1071:1077 Double mutants 1064:1077 Double mutants F571L/N574S and F571L/N574A 1064:1105 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 7 42 dep mutants 1071:1077 arg1 F571L/N574A 1095:1105 F571L/N574A 1095:1105 F571L/N574A 1095:1105 Double mutants F571L/N574S and F571L/N574A showed an increase of about 2-fold. 26998654 1 43 used used 197:200 arg2 hydrolases 147:156 Glycoside hydrolases 137:156 Glycoside hydrolases 137:156 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 2 44 with variants 503:510 arg1 performance 524:534 better performance 517:534 better performance for GOS biosynthesis 517:555 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 0 45 theme maritima 55:62 arg1 Site 36:39 the Active Site 25:39 the Active Site of Thermotoga maritima 25:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 0 45 theme maritima 55:62 arg1 maritima 55:62 Thermotoga maritima 44:62 Thermotoga maritima 44:62 Structural Dissection of the Active Site of Thermotoga maritima β-Galactosidase Identifies Key Residues for Transglycosylating Activity. 26998654 4 46 theme wild-type 853:861 arg1 enzyme 863:868 the wild-type enzyme 849:868 the wild-type enzyme 849:868 Analysis of mutant enzymes indicated that residue W959, or an alternative aromatic residue at this position, is critical for the synthesis of β-3'-galactosyl-lactose, the major GOS obtained with the wild-type enzyme. 26998654 1 47 theme β-galactosidases 172:187 arg1 hydrolases 147:156 Glycoside hydrolases 137:156 Glycoside hydrolases 137:156 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 26998654 2 48 from mutagenesis 330:340 arg1 maritima 395:402 Thermotoga maritima 384:402 Thermotoga maritima 384:402 Site-directed mutagenesis of a thermoresistant β-galactosidase from Thermotoga maritima has been carried out to study the structural basis of transgalactosylation and to obtain enzymatic variants with better performance for GOS biosynthesis. 26998654 1 49 theme transglycosylating 258:275 arg1 activity 289:296 the transglycosylating (secondary) activity 254:296 the transglycosylating (secondary) activity of these enzymes 254:313 Glycoside hydrolases, specifically β-galactosidases, can be used to synthesize galacto-oligosaccharides (GOS) due to the transglycosylating (secondary) activity of these enzymes. 29124306 5 0 theme subsequent 651:660 arg1 cycloaddition 662:674 subsequent cycloaddition 651:674 subsequent cycloaddition of an alkyne 651:687 This azide can be used for subsequent cycloaddition of an alkyne. 29124306 6 1 theme macrocyclic 738:748 arg1 reagent 750:756 a lanthanide-containing macrocyclic reagent 714:756 a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose 714:803 Therefore, we developed a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose. 29124306 1 2 theme modern 137:142 arg1 chemistry 158:166 modern bioanalytical chemistry 137:166 modern bioanalytical chemistry 137:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 2 3 from possibilities 294:306 arg1 analyses 330:337 mass spectrometric analyses 311:337 mass spectrometric analyses 311:337 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 4 4 theme galactose 595:603 arg1 residue 605:611 galactose residue 595:611 galactose residue 595:611 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 10 5 with labeling 1305:1312 arg1 elements 1348:1355 rare earth elements 1337:1355 rare earth elements 1337:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 1 6 theme bioanalytical 144:156 arg1 chemistry 158:166 modern bioanalytical chemistry 137:166 modern bioanalytical chemistry 137:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 7 7 theme metal 877:881 arg1 functionality 883:895 a metal functionality 875:895 a metal functionality using this approach 875:915 In summary, the glycan structures of an antibody can be labeled with a metal functionality using this approach. 29124306 8 8 theme labeling 1035:1042 arg1 reagents 1044:1051 labeling reagents 1035:1051 labeling reagents that target amino or thiol groups 1035:1085 Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. 29124306 9 9 theme quantitative 1187:1198 arg1 analyses 1200:1207 quantitative analyses 1187:1207 quantitative analyses 1187:1207 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 2 10 theme numerous 285:292 arg1 possibilities 294:306 numerous possibilities 285:306 numerous possibilities in mass spectrometric analyses 285:337 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 10 11 theme elemental 1382:1390 arg1 spectrometry 1397:1408 elemental mass spectrometry 1382:1408 elemental mass spectrometry 1382:1408 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 3 12 theme labeling 384:391 arg1 strategy 393:400 a new metal labeling strategy 372:400 a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies 372:467 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 1 13 theme Quantitative 70:81 arg1 analysis 83:90 Quantitative analysis 70:90 Quantitative analysis of complex proteins 70:110 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 1 13 theme Quantitative 70:81 arg1 task 129:132 a challenging task 115:132 a challenging task in modern bioanalytical chemistry 115:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 1 14 from task 129:132 arg1 chemistry 158:166 modern bioanalytical chemistry 137:166 modern bioanalytical chemistry 137:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 6 15 theme lanthanide-containing 716:736 arg1 reagent 750:756 a lanthanide-containing macrocyclic reagent 714:756 a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose 714:803 Therefore, we developed a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose. 29124306 9 16 from experiments 1220:1230 arg1 samples 1268:1274 complex biological samples 1249:1274 complex biological samples 1249:1274 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 8 17 theme amino 1065:1069 arg1 groups 1080:1085 amino or thiol groups 1065:1085 groups 1080:1085 Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. 29124306 10 18 theme abstract 1287:1294 arg1 labeling 1305:1312 Graphical abstract Antibody labeling 1277:1312 Graphical abstract Antibody labeling at sugar moieties with rare earth elements 1277:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 10 19 theme sugar 1317:1321 arg1 moieties 1323:1330 sugar moieties 1317:1330 sugar moieties 1317:1330 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 10 20 from application 1367:1377 arg1 spectrometry 1397:1408 elemental mass spectrometry 1382:1408 elemental mass spectrometry 1382:1408 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 0 21 from labeling 25:32 arg1 antibodies 58:67 antibodies 58:67 antibodies 58:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 9 22 theme imaging 1212:1218 arg1 experiments 1220:1230 imaging experiments 1212:1230 imaging experiments 1212:1230 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 2 23 theme metal 267:271 arg1 labels 273:278 new metal labels 263:278 new metal labels 263:278 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 3 24 theme proteins 430:437 arg1 structures 416:425 tag glycan structures 405:425 tag glycan structures of proteins 405:437 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 0 25 theme new 2:4 arg1 strategy 6:13 A new strategy 0:13 A new strategy for metal labeling of glycan structures in antibodies 0:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 4 26 theme oligosaccharide 474:488 arg1 glycans 490:496 The oligosaccharide glycans 470:496 The oligosaccharide glycans 470:496 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 8 27 theme thiol 1074:1078 arg1 groups 1080:1085 amino or thiol groups 1065:1085 groups 1080:1085 Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. 29124306 8 28 theme Fc 1007:1008 arg1 glycans 1010:1016 the Fc glycans 1003:1016 the Fc glycans instead of using labeling reagents that target amino or thiol groups 1003:1085 Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. 29124306 2 29 theme spectrometric 316:328 arg1 analyses 330:337 mass spectrometric analyses 311:337 mass spectrometric analyses 311:337 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 3 30 theme glycan 409:414 arg1 structures 416:425 tag glycan structures 405:425 tag glycan structures of proteins 405:437 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 3 31 theme metal 378:382 arg1 strategy 393:400 a new metal labeling strategy 372:400 a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies 372:467 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 7 32 theme glycan 822:827 arg1 structures 829:838 the glycan structures 818:838 the glycan structures of an antibody 818:853 In summary, the glycan structures of an antibody can be labeled with a metal functionality using this approach. 29124306 2 33 theme mass 311:314 arg1 analyses 330:337 mass spectrometric analyses 311:337 mass spectrometric analyses 311:337 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 7 34 theme antibody 846:853 arg1 structures 829:838 the glycan structures 818:838 the glycan structures of an antibody 818:853 In summary, the glycan structures of an antibody can be labeled with a metal functionality using this approach. 29124306 9 35 theme complex 1249:1255 arg1 samples 1268:1274 complex biological samples 1249:1274 complex biological samples 1249:1274 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 10 36 from moieties 1323:1330 arg1 labeling 1305:1312 Graphical abstract Antibody labeling 1277:1312 Graphical abstract Antibody labeling at sugar moieties with rare earth elements 1277:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 9 37 theme antibodies 1235:1244 arg1 analyses 1200:1207 quantitative analyses 1187:1207 quantitative analyses 1187:1207 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 9 37 theme antibodies 1235:1244 arg1 experiments 1220:1230 imaging experiments 1212:1230 imaging experiments 1212:1230 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 10 38 theme earth 1342:1346 arg1 elements 1348:1355 rare earth elements 1337:1355 rare earth elements 1337:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 9 39 theme mass 1164:1167 arg1 spectrometry 1169:1180 molecular mass spectrometry 1154:1180 molecular mass spectrometry 1154:1180 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 2 40 theme available 178:186 arg1 labels 196:201 Commonly available isotope labels 169:201 Commonly available isotope labels 169:201 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 1 41 theme complex 95:101 arg1 proteins 103:110 complex proteins 95:110 complex proteins 95:110 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 10 42 theme rare 1337:1340 arg1 earth 1342:1346 rare earth 1337:1346 rare earth elements 1337:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 2 43 dep suffering 213:221 arg1 whereas 255:261 whereas 255:261 whereas 255:261 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 4 44 theme artificial 567:576 arg1 azide 578:582 an artificial azide 564:582 an artificial azide containing galactose residue 564:611 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 0 45 from antibodies 58:67 arg1 labeling 25:32 metal labeling 19:32 metal labeling of glycan structures in antibodies 19:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 5 46 used used 642:645 arg2 azide 629:633 This azide 624:633 This azide 624:633 This azide can be used for subsequent cycloaddition of an alkyne. 29124306 0 47 theme metal 19:23 arg1 labeling 25:32 metal labeling 19:32 metal labeling of glycan structures in antibodies 19:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 4 48 theme last 530:533 arg1 N-acetylglucosamine 535:553 the last N-acetylglucosamine 526:553 the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound 526:621 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 3 49 theme tag 405:407 arg1 structures 416:425 tag glycan structures 405:425 tag glycan structures of proteins 405:437 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 9 50 theme biological 1257:1266 arg1 samples 1268:1274 complex biological samples 1249:1274 complex biological samples 1249:1274 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 3 51 dep structures 416:425 arg1 antibodies 458:467 antibodies 458:467 antibodies 458:467 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 6 52 theme azido 789:793 arg1 galactose 795:803 this azido galactose 784:803 this azido galactose 784:803 Therefore, we developed a lanthanide-containing macrocyclic reagent to selectively connect to this azido galactose. 29124306 0 53 theme glycan 37:42 arg1 structures 44:53 glycan structures 37:53 glycan structures in antibodies 37:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 5 54 theme alkyne 682:687 arg1 cycloaddition 662:674 subsequent cycloaddition 651:674 subsequent cycloaddition of an alkyne 651:687 This azide can be used for subsequent cycloaddition of an alkyne. 29124306 0 55 from structures 44:53 arg1 antibodies 58:67 antibodies 58:67 antibodies 58:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 4 56 contain containing 584:593 arg2 residue 605:611 galactose residue 595:611 galactose residue 595:611 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 4 56 contain containing 584:593 arg1 azide 578:582 an artificial azide 564:582 an artificial azide containing galactose residue 564:611 The oligosaccharide glycans were selectively trimmed to the last N-acetylglucosamine to which an artificial azide containing galactose residue was bound. 29124306 9 57 theme elemental 1135:1143 arg1 spectrometry 1169:1180 molecular mass spectrometry 1154:1180 molecular mass spectrometry 1154:1180 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 1 58 theme proteins 103:110 arg1 analysis 83:90 Quantitative analysis 70:90 Quantitative analysis of complex proteins 70:110 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 1 58 theme proteins 103:110 arg1 task 129:132 a challenging task 115:132 a challenging task in modern bioanalytical chemistry 115:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 9 59 from antibodies 1235:1244 arg1 samples 1268:1274 complex biological samples 1249:1274 complex biological samples 1249:1274 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 2 60 theme new 263:265 arg1 labels 273:278 new metal labels 263:278 new metal labels 263:278 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 10 61 theme Graphical 1277:1285 arg1 labeling 1305:1312 Graphical abstract Antibody labeling 1277:1312 Graphical abstract Antibody labeling at sugar moieties with rare earth elements 1277:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 9 62 theme molecular 1154:1162 arg1 spectrometry 1169:1180 molecular mass spectrometry 1154:1180 molecular mass spectrometry 1154:1180 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 1 63 theme challenging 117:127 arg1 analysis 83:90 Quantitative analysis 70:90 Quantitative analysis of complex proteins 70:110 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 1 63 theme challenging 117:127 arg1 task 129:132 a challenging task 115:132 a challenging task in modern bioanalytical chemistry 115:166 Quantitative analysis of complex proteins is a challenging task in modern bioanalytical chemistry. 29124306 2 64 theme isotope 188:194 arg1 labels 196:201 Commonly available isotope labels 169:201 Commonly available isotope labels 169:201 Commonly available isotope labels are still suffering from limitations and drawbacks, whereas new metal labels open numerous possibilities in mass spectrometric analyses. 29124306 8 65 theme antibodies 956:965 arg1 functionality 935:947 the functionality 931:947 the functionality of the antibodies 931:965 Furthermore, the functionality of the antibodies can be fully maintained by labeling the Fc glycans instead of using labeling reagents that target amino or thiol groups. 29124306 9 66 from analyses 1200:1207 arg1 samples 1268:1274 complex biological samples 1249:1274 complex biological samples 1249:1274 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 0 67 theme structures 44:53 arg1 labeling 25:32 metal labeling 19:32 metal labeling of glycan structures in antibodies 19:67 A new strategy for metal labeling of glycan structures in antibodies. 29124306 10 68 theme mass 1392:1395 arg1 spectrometry 1397:1408 elemental mass spectrometry 1382:1408 elemental mass spectrometry 1382:1408 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 3 69 theme new 374:376 arg1 strategy 393:400 a new metal labeling strategy 372:400 a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies 372:467 In this work, we have developed a new metal labeling strategy to tag glycan structures of proteins, more particularly antibodies. 29124306 10 70 theme Antibody 1296:1303 arg1 labeling 1305:1312 Graphical abstract Antibody labeling 1277:1312 Graphical abstract Antibody labeling at sugar moieties with rare earth elements 1277:1355 Graphical abstract Antibody labeling at sugar moieties with rare earth elements to enable application in elemental mass spectrometry. 29124306 9 71 from samples 1268:1274 arg1 analyses 1200:1207 quantitative analyses 1187:1207 quantitative analyses 1187:1207 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 29124306 9 71 from samples 1268:1274 arg1 experiments 1220:1230 imaging experiments 1212:1230 imaging experiments 1212:1230 This approach enables the possibility of using elemental, besides molecular mass spectrometry, for quantitative analyses or imaging experiments of antibodies in complex biological samples. 25732693 0 0 theme pancreatic 215:224 arg1 diagnosis 234:242 pancreatic disease diagnosis 215:242 pancreatic disease diagnosis 215:242 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 1 1 theme capillary 413:421 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 1 theme capillary 413:421 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 8 2 theme increases 1720:1728 arg1 stage 1699:1703 the stage 1695:1703 the stage of the disease increases 1695:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 1 3 theme reductive 297:305 arg1 GRIL 325:328 GRIL 325:328 GRIL 325:328 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 3 theme reductive 297:305 arg1 labeling 315:322 glycan reductive isotope labeling 290:322 glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline 290:370 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 2 4 theme Human 615:619 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 4 theme Human 615:619 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 4 theme Human 615:619 arg1 hAGP 643:646 hAGP 643:646 hAGP 643:646 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 4 5 theme chronic 1058:1064 arg1 pancreatitis 1066:1077 chronic pancreatitis 1058:1077 chronic pancreatitis 1058:1077 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 8 6 theme fucosylated 1672:1682 arg1 glycans 1684:1690 fucosylated glycans 1672:1690 fucosylated glycans as the stage of the disease increases 1672:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 6 7 theme denaturing 1297:1306 arg1 conditions 1308:1317 protein denaturing conditions 1289:1317 protein denaturing conditions prior to PNGase F digestion 1289:1345 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 1 8 theme chromatography 430:443 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 8 theme chromatography 430:443 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 5 9 theme isotope 1233:1239 arg1 labeling 1241:1248 stable isotope labeling 1226:1248 stable isotope labeling 1226:1248 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 7 10 theme proposed 1503:1510 arg1 method 1512:1517 the proposed method 1499:1517 the proposed method 1499:1517 Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). 25732693 9 11 theme more 1931:1934 arg1 cases 1945:1949 more clinical cases 1931:1949 more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer 1931:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 1 12 theme mass 458:461 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 12 theme mass 458:461 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 13 from glycan 1835:1840 arg1 hAGP 1845:1848 hAGP 1845:1848 hAGP 1845:1848 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 10 14 theme protein 2140:2146 arg1 enrichment 2117:2126 an enrichment 2114:2126 an enrichment of a target protein by IAC 2114:2153 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 4 15 theme pancreatic 1103:1112 arg1 cancer 1114:1119 pancreatic cancer 1103:1119 pancreatic cancer 1103:1119 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 0 16 theme liquid 150:155 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 1 17 theme C 355:355 arg1 aniline 364:370 [(13)C]-coded aniline 350:370 [(13)C]-coded aniline 350:370 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 17 theme C 355:355 arg1 13 352:353 13 352:353 13 352:353 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 8 18 theme chronic 1766:1772 arg1 pancreatitis 1774:1785 chronic pancreatitis 1766:1785 chronic pancreatitis 1766:1785 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 11 19 theme associated 2369:2378 arg1 biomarkers 2393:2402 novel cancer associated glycoprotein biomarkers 2356:2402 novel cancer associated glycoprotein biomarkers 2356:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 10 20 theme isotope 2160:2166 arg1 quantitation 2183:2194 isotope coded relative quantitation 2160:2194 isotope coded relative quantitation of N-glycans 2160:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 2 21 gly glycoprotein 673:684 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 21 gly glycoprotein 673:684 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 1 22 theme -coded 357:362 arg1 aniline 364:370 [(13)C]-coded aniline 350:370 [(13)C]-coded aniline 350:370 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 22 theme -coded 357:362 arg1 13 352:353 13 352:353 13 352:353 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 6 23 theme standard 1478:1485 arg1 hAGP 1487:1490 standard hAGP 1478:1490 standard hAGP 1478:1490 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 1 24 from samples 585:591 arg1 present 574:580 present 574:580 present 574:580 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 25 from present 574:580 arg1 samples 585:591 samples 585:591 samples from cancer patients 585:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 26 theme initial 1999:2005 arg1 stages 2007:2012 initial stages 1999:2012 initial stages of pancreatic cancer 1999:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 10 27 used used 2229:2232 arg2 methodology 2092:2102 the presented methodology 2078:2102 the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans 2078:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 6 28 theme PNGase 1328:1333 arg1 digestion 1337:1345 PNGase F digestion 1328:1345 PNGase F digestion 1328:1345 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 1 29 theme hydrophilic 389:399 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 29 theme hydrophilic 389:399 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 11 30 theme glycoprotein 2380:2391 arg1 biomarkers 2393:2402 novel cancer associated glycoprotein biomarkers 2356:2402 novel cancer associated glycoprotein biomarkers 2356:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 1 31 theme isotope 307:313 arg1 GRIL 325:328 GRIL 325:328 GRIL 325:328 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 31 theme isotope 307:313 arg1 labeling 315:322 glycan reductive isotope labeling 290:322 glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline 290:370 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 32 theme cancer 2028:2033 arg1 stages 2007:2012 initial stages 1999:2012 initial stages of pancreatic cancer 1999:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 2 33 theme serum 667:671 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 33 theme serum 667:671 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 0 34 theme zwitterionic 103:114 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 4 35 theme immunoaffinity 955:968 arg1 IAC 986:988 IAC 986:988 IAC 986:988 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 4 35 theme immunoaffinity 955:968 arg1 chromatography 970:983 immunoaffinity chromatography 955:983 immunoaffinity chromatography (IAC) 955:989 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 0 36 from alfa-acid-glycoprotein 46:67 arg1 N-glycans 25:33 N-glycans 25:33 N-glycans from human alfa-acid-glycoprotein 25:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 0 36 from alfa-acid-glycoprotein 46:67 arg1 analysis 13:20 Quantitative analysis 0:20 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein 0:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 2 37 theme acute 655:659 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 37 theme acute 655:659 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 1 38 theme labeling 315:322 arg1 potential 277:285 the potential 273:285 the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients 273:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 4 39 theme individuals 1021:1031 arg1 samples 1002:1008 serum samples 996:1008 serum samples of healthy individuals 996:1031 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 0 40 theme Quantitative 0:11 arg1 analysis 13:20 Quantitative analysis 0:20 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein 0:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 5 41 theme hAGP 1191:1194 arg1 glycans 1196:1202 the hAGP glycans 1187:1202 the hAGP glycans 1187:1202 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 3 42 from glycans 828:834 arg1 hAGP 839:842 hAGP 839:842 hAGP 839:842 However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. 25732693 1 43 from quantitation 509:520 arg1 glycoproteins 560:572 selected glycoproteins 551:572 selected glycoproteins present in samples from cancer patients 551:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 4 44 theme serum 996:1000 arg1 samples 1002:1008 serum samples 996:1008 serum samples of healthy individuals 996:1031 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 5 45 theme relative 1162:1169 arg1 quantitation 1171:1182 relative quantitation 1162:1182 relative quantitation of the hAGP glycans 1162:1202 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 7 46 theme clinical 1554:1561 arg1 samples 1563:1569 the clinical samples 1550:1569 the clinical samples (control vs. pathological) 1550:1596 Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). 25732693 8 47 theme Pancreatic 1599:1608 arg1 samples 1617:1623 Pancreatic cancer samples 1599:1623 Pancreatic cancer samples 1599:1623 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 0 48 theme stable 75:80 arg1 labeling 90:97 stable isotope labeling 75:97 stable isotope labeling 75:97 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 6 49 theme established 1435:1445 arg1 methodology 1447:1457 the established methodology 1431:1457 the established methodology 1431:1457 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 4 50 theme different 1083:1091 arg1 stages 1093:1098 different stages 1083:1098 different stages of pancreatic cancer 1083:1119 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 8 51 theme glycans 1684:1690 arg1 abundance 1659:1667 the abundance 1655:1667 the abundance of fucosylated glycans as the stage of the disease increases 1655:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 1 52 theme glycosylation 525:537 arg1 variants 539:546 glycosylation variants 525:546 glycosylation variants 525:546 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 5 53 theme μZIC-HILIC-ESI-MS 1254:1270 arg1 analysis 1272:1279 μZIC-HILIC-ESI-MS analysis 1254:1279 μZIC-HILIC-ESI-MS analysis 1254:1279 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 1 54 gly glycoproteins 560:572 arg1 glycoproteins 560:572 selected glycoproteins 551:572 selected glycoproteins present in samples from cancer patients 551:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 8 55 gly fucosylated 1672:1682 arg1 glycans 1684:1690 fucosylated glycans 1672:1690 fucosylated glycans as the stage of the disease increases 1672:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 10 56 theme target 2133:2138 arg1 protein 2140:2146 a target protein 2131:2146 a target protein 2131:2146 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 0 57 theme disease 226:232 arg1 diagnosis 234:242 pancreatic disease diagnosis 215:242 pancreatic disease diagnosis 215:242 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 10 58 theme targeted 2238:2245 arg1 studies 2257:2263 targeted glycomics studies 2238:2263 targeted glycomics studies 2238:2263 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 1 59 theme glycan 290:295 arg1 GRIL 325:328 GRIL 325:328 GRIL 325:328 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 59 theme glycan 290:295 arg1 labeling 315:322 glycan reductive isotope labeling 290:322 glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline 290:370 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 60 theme extended 1906:1913 arg1 study 1915:1919 an extended study 1903:1919 an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer 1903:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 1 61 theme liquid 423:428 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 61 theme liquid 423:428 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 8 62 theme disease 1712:1718 arg1 increases 1720:1728 the disease increases 1708:1728 the disease increases 1708:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 5 63 theme stable 1226:1231 arg1 labeling 1241:1248 stable isotope labeling 1226:1248 stable isotope labeling 1226:1248 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 6 64 theme protein 1289:1295 arg1 conditions 1308:1317 protein denaturing conditions 1289:1317 protein denaturing conditions prior to PNGase F digestion 1289:1345 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 1 65 theme electrospray 445:456 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 65 theme electrospray 445:456 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 6 66 theme digestion 1386:1394 arg1 yields 1396:1401 quantitative digestion yields 1373:1401 quantitative digestion yields 1373:1401 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 9 67 theme clinical 1936:1943 arg1 cases 1945:1949 more clinical cases 1931:1949 more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer 1931:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 1 68 theme spectrometry 463:474 arg1 potential 277:285 the potential 273:285 the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients 273:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 4 69 theme cancer 1114:1119 arg1 pancreatitis 1066:1077 chronic pancreatitis 1058:1077 chronic pancreatitis 1058:1077 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 4 69 theme cancer 1114:1119 arg1 stages 1093:1098 different stages 1083:1098 different stages of pancreatic cancer 1083:1119 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 8 70 from pancreatitis 1774:1785 arg1 samples 1753:1759 samples 1753:1759 samples from chronic pancreatitis 1753:1785 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 6 71 theme quantitative 1373:1384 arg1 yields 1396:1401 quantitative digestion yields 1373:1401 quantitative digestion yields 1373:1401 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 2 72 gly α1-acid-glycoprotein 621:640 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 72 gly α1-acid-glycoprotein 621:640 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 72 gly α1-acid-glycoprotein 621:640 arg1 hAGP 643:646 hAGP 643:646 hAGP 643:646 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 10 73 theme coded 2168:2172 arg1 quantitation 2183:2194 isotope coded relative quantitation 2160:2194 isotope coded relative quantitation of N-glycans 2160:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 0 74 theme mass 185:188 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 11 75 theme novel 2356:2360 arg1 biomarkers 2393:2402 novel cancer associated glycoprotein biomarkers 2356:2402 novel cancer associated glycoprotein biomarkers 2356:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 1 76 attach present 574:580 arg1 samples 585:591 samples 585:591 samples from cancer patients 585:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 76 attach present 574:580 arg2 glycoproteins 560:572 selected glycoproteins 551:572 selected glycoproteins present in samples from cancer patients 551:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 77 from patients 605:612 arg1 samples 585:591 samples 585:591 samples from cancer patients 585:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 3 78 used used 851:854 arg2 biomarker 859:867 biomarker 859:867 biomarker 859:867 However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. 25732693 3 78 used used 851:854 arg2 glycans 828:834 some particular glycans 812:834 some particular glycans in hAGP 812:842 However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. 25732693 9 79 dep cases 1945:1949 arg1 those 1963:1967 those 1963:1967 those 1963:1967 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 9 79 dep cases 1945:1949 arg1 stages 2007:2012 initial stages 1999:2012 initial stages of pancreatic cancer 1999:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 0 80 theme chromatography 157:170 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 10 81 theme N-glycans 2199:2207 arg1 quantitation 2183:2194 isotope coded relative quantitation 2160:2194 isotope coded relative quantitation of N-glycans 2160:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 6 82 theme F 1335:1335 arg1 digestion 1337:1345 PNGase F digestion 1328:1345 PNGase F digestion 1328:1345 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 1 83 theme zwitterionic 376:387 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 83 theme zwitterionic 376:387 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 2 84 theme chronic 753:759 arg1 inflammation 761:772 chronic inflammation 753:772 chronic inflammation 753:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 10 85 theme relative 2174:2181 arg1 quantitation 2183:2194 isotope coded relative quantitation 2160:2194 isotope coded relative quantitation of N-glycans 2160:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 3 86 theme particular 817:826 arg1 biomarker 859:867 biomarker 859:867 biomarker 859:867 However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. 25732693 3 86 theme particular 817:826 arg1 glycans 828:834 some particular glycans 812:834 some particular glycans in hAGP 812:842 However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. 25732693 0 87 theme isotope 82:88 arg1 labeling 90:97 stable isotope labeling 75:97 stable isotope labeling 75:97 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 1 88 theme interaction 401:411 arg1 spectrometry 463:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 376:474 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) 376:494 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 1 88 theme interaction 401:411 arg1 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 μZIC-HILIC-ESI-MS 477:493 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 89 theme pancreatic 2017:2026 arg1 cancer 2028:2033 pancreatic cancer 2017:2033 pancreatic cancer 2017:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 8 90 from increase 1643:1650 arg1 abundance 1659:1667 the abundance 1655:1667 the abundance of fucosylated glycans as the stage of the disease increases 1655:1728 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 0 91 from analysis 13:20 arg1 alfa-acid-glycoprotein 46:67 human alfa-acid-glycoprotein 40:67 human alfa-acid-glycoprotein 40:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 0 92 theme hydrophilic 116:126 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 2 93 theme phase 661:665 arg1 α1-acid-glycoprotein 621:640 Human α1-acid-glycoprotein 615:640 Human α1-acid-glycoprotein (hAGP) 615:647 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 93 theme phase 661:665 arg1 glycoprotein 673:684 an acute phase serum glycoprotein 652:684 an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation 652:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 11 94 theme other 2320:2324 arg1 studies 2331:2337 other such studies 2320:2337 other such studies aimed at finding novel cancer associated glycoprotein biomarkers 2320:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 1 95 theme selected 551:558 arg1 glycoproteins 560:572 selected glycoproteins 551:572 selected glycoproteins present in samples from cancer patients 551:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 0 96 theme capillary 140:148 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 1 97 theme present 574:580 arg1 glycoproteins 560:572 selected glycoproteins 551:572 selected glycoproteins present in samples from cancer patients 551:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 4 98 theme healthy 1013:1019 arg1 individuals 1021:1031 healthy individuals 1013:1031 healthy individuals 1013:1031 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 0 99 theme N-glycans 25:33 arg1 analysis 13:20 Quantitative analysis 0:20 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein 0:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 5 100 theme glycans 1196:1202 arg1 quantitation 1171:1182 relative quantitation 1162:1182 relative quantitation of the hAGP glycans 1162:1202 After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and μZIC-HILIC-ESI-MS analysis. 25732693 10 101 theme glycomics 2247:2255 arg1 studies 2257:2263 targeted glycomics studies 2238:2263 targeted glycomics studies 2238:2263 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 11 102 theme such 2326:2329 arg1 studies 2331:2337 other such studies 2320:2337 other such studies aimed at finding novel cancer associated glycoprotein biomarkers 2320:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 1 103 theme cancer 598:603 arg1 patients 605:612 cancer patients 598:612 cancer patients 598:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 0 104 theme human 40:44 arg1 alfa-acid-glycoprotein 46:67 human alfa-acid-glycoprotein 40:67 human alfa-acid-glycoprotein 40:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 11 105 gly glycoprotein 2380:2391 arg1 glycoprotein 2380:2391 novel cancer associated glycoprotein biomarkers 2356:2402 novel cancer associated glycoprotein biomarkers 2356:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 8 106 theme cancer 1610:1615 arg1 samples 1617:1623 Pancreatic cancer samples 1599:1623 Pancreatic cancer samples 1599:1623 Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. 25732693 0 107 theme interaction 128:138 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 6 108 theme methodology 1447:1457 arg1 reproducibility 1412:1426 the reproducibility 1408:1426 the reproducibility of the established methodology 1408:1457 First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. 25732693 7 109 theme samples 1563:1569 arg1 analysis 1538:1545 the analysis 1534:1545 the analysis of the clinical samples (control vs. pathological) 1534:1596 Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). 25732693 0 110 theme electrospray 172:183 arg1 spectrometry 190:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry 103:201 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 1 111 theme relative 500:507 arg1 quantitation 509:520 relative quantitation 500:520 relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients 500:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 11 112 theme cancer 2362:2367 arg1 biomarkers 2393:2402 novel cancer associated glycoprotein biomarkers 2356:2402 novel cancer associated glycoprotein biomarkers 2356:2402 The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. 25732693 9 113 theme chronic 1974:1980 arg1 pancreatitis 1982:1993 chronic pancreatitis 1974:1993 chronic pancreatitis 1974:1993 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 7 114 dep samples 1563:1569 arg1 pathological 1584:1595 pathological 1584:1595 pathological 1584:1595 Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). 25732693 7 114 dep samples 1563:1569 arg1 control 1572:1578 control 1572:1578 control 1572:1578 Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). 25732693 0 115 gly alfa-acid-glycoprotein 46:67 arg1 alfa-acid-glycoprotein 46:67 human alfa-acid-glycoprotein 40:67 human alfa-acid-glycoprotein 40:67 Quantitative analysis of N-glycans from human alfa-acid-glycoprotein using stable isotope labeling and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry as tool for pancreatic disease diagnosis. 25732693 4 116 attach isolated 943:950 arg2 hAGP 934:937 hAGP 934:937 hAGP 934:937 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 4 116 attach isolated 943:950 arg1 samples 1002:1008 serum samples 996:1008 serum samples of healthy individuals 996:1031 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 4 116 attach isolated 943:950 arg1 those 1042:1046 those 1042:1046 those 1042:1046 In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. 25732693 10 117 theme presented 2082:2090 arg1 methodology 2092:2102 the presented methodology 2078:2102 the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans 2078:2207 Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. 25732693 1 118 theme variants 539:546 arg1 quantitation 509:520 relative quantitation 500:520 relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients 500:612 In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (μZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. 25732693 9 119 theme mentioned 1825:1833 arg1 candidate 1855:1863 a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer 1853:2033 a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer 1853:2033 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 9 119 theme mentioned 1825:1833 arg1 glycan 1835:1840 the mentioned glycan 1821:1840 the mentioned glycan in hAGP 1821:1848 The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. 25732693 2 120 gly glycosylation 692:704 arg1 cancer 742:747 cancer 742:747 cancer 742:747 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25732693 2 120 gly glycosylation 692:704 arg1 inflammation 761:772 chronic inflammation 753:772 chronic inflammation 753:772 Human α1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. 25497333 1 0 theme complex 175:181 arg1 polysaccharides 199:213 structurally complex plant cell wall polysaccharides 162:213 structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling 162:294 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 0 theme complex 175:181 arg1 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 1 theme energy 263:268 arg1 metabolism 270:279 energy metabolism 263:279 energy metabolism 263:279 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 0 2 theme oligosaccharides 80:95 arg1 library 42:48 a library 40:48 a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature 40:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 1 3 theme plant 183:187 arg1 polysaccharides 199:213 structurally complex plant cell wall polysaccharides 162:213 structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling 162:294 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 3 theme plant 183:187 arg1 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 3 4 theme chromatography 722:735 arg1 combination 654:664 a combination 652:664 a combination of enzymatic and chemical treatments and size-exclusion chromatography 652:735 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 3 5 theme acidic 611:616 arg1 oligosaccharides 629:644 neutral and acidic xyloglucan oligosaccharides 599:644 neutral and acidic xyloglucan oligosaccharides 599:644 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 4 6 theme matrix-assisted 863:877 arg1 spectrometry 912:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 6 7 theme correlation 1197:1207 arg1 spectroscopy 1209:1220 two-dimensional correlation spectroscopy 1181:1220 two-dimensional correlation spectroscopy 1181:1220 1H chemical shifts were assigned using two-dimensional correlation spectroscopy. 25497333 3 8 dep generation 568:577 arg1 the 541:543 the 541:543 the 541:543 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 4 9 theme desorption/ionization 885:905 arg1 spectrometry 912:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 3 10 theme xyloglucan 618:627 arg1 oligosaccharides 629:644 neutral and acidic xyloglucan oligosaccharides 599:644 neutral and acidic xyloglucan oligosaccharides 599:644 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 5 11 theme Mass 1022:1025 arg1 spectrometry 1027:1038 Mass spectrometry 1022:1038 Mass spectrometry 1022:1038 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 5 11 theme Mass 1022:1025 arg1 chromatography 1055:1068 analytical chromatography 1044:1068 analytical chromatography 1044:1068 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 5 11 theme Mass 1022:1025 arg1 methods 1087:1093 methods 1087:1093 methods for xyloglucan oligosaccharide quantification 1087:1139 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 4 12 theme resonance 998:1006 arg1 spectroscopy 1008:1019 1H nuclear magnetic resonance spectroscopy 978:1019 1H nuclear magnetic resonance spectroscopy 978:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 3 13 theme neutral 599:605 arg1 oligosaccharides 629:644 neutral and acidic xyloglucan oligosaccharides 599:644 neutral and acidic xyloglucan oligosaccharides 599:644 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 1 14 theme cell 189:192 arg1 polysaccharides 199:213 structurally complex plant cell wall polysaccharides 162:213 structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling 162:294 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 14 theme cell 189:192 arg1 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 6 15 theme two-dimensional 1181:1195 arg1 spectroscopy 1209:1220 two-dimensional correlation spectroscopy 1181:1220 two-dimensional correlation spectroscopy 1181:1220 1H chemical shifts were assigned using two-dimensional correlation spectroscopy. 25497333 4 16 theme magnetic 989:996 arg1 resonance 998:1006 1H nuclear magnetic resonance 978:1006 1H nuclear magnetic resonance spectroscopy 978:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 4 17 theme spectrometry 912:923 arg1 combination 848:858 a combination 846:858 a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy 846:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 1 18 theme wall 194:197 arg1 polysaccharides 199:213 structurally complex plant cell wall polysaccharides 162:213 structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling 162:294 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 18 theme wall 194:197 arg1 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans 146:156 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 4 19 theme oligosaccharides 758:773 arg1 Twenty-six 738:747 Twenty-six 738:747 Twenty-six 738:747 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 4 19 theme oligosaccharides 758:773 arg1 oligosaccharides 758:773 these oligosaccharides 752:773 these oligosaccharides 752:773 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 2 20 theme xyloglucans 349:359 arg1 relationships 332:344 the structure-function relationships 309:344 the structure-function relationships of xyloglucans 309:359 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 4 21 theme high-performance 926:941 arg1 chromatography 958:971 high-performance anion exchange chromatography 926:971 high-performance anion exchange chromatography 926:971 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 4 22 theme spectroscopy 1008:1019 arg1 combination 848:858 a combination 846:858 a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy 846:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 4 23 theme mass 907:910 arg1 spectrometry 912:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 5 24 theme analytical 1044:1053 arg1 spectrometry 1027:1038 Mass spectrometry 1022:1038 Mass spectrometry 1022:1038 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 5 24 theme analytical 1044:1053 arg1 chromatography 1055:1068 analytical chromatography 1044:1068 analytical chromatography 1044:1068 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 5 24 theme analytical 1044:1053 arg1 methods 1087:1093 methods 1087:1093 methods for xyloglucan oligosaccharide quantification 1087:1139 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 4 25 theme chromatography 958:971 arg1 combination 848:858 a combination 846:858 a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy 846:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 4 26 theme 1H 978:979 arg1 resonance 998:1006 1H nuclear magnetic resonance 978:1006 1H nuclear magnetic resonance spectroscopy 978:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 3 27 theme oligosaccharides 629:644 arg1 purification 583:594 purification 583:594 purification 583:594 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 3 27 theme oligosaccharides 629:644 arg1 generation 568:577 semi-preparative scale generation 545:577 semi-preparative scale generation 545:577 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 2 28 theme xyloglucan 480:489 arg1 oligosaccharides 491:506 rigorously characterized xyloglucan oligosaccharides 455:506 rigorously characterized xyloglucan oligosaccharides 455:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 2 29 theme oligosaccharides 491:506 arg1 collection 441:450 a comprehensive and structurally diverse collection 400:450 a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides 400:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 0 30 theme xyloglucan 121:130 arg1 nomenclature 132:143 xyloglucan nomenclature 121:143 xyloglucan nomenclature 121:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 2 31 theme structure-function 313:330 arg1 relationships 332:344 the structure-function relationships 309:344 the structure-function relationships of xyloglucans 309:359 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 2 32 theme characterized 466:478 arg1 oligosaccharides 491:506 rigorously characterized xyloglucan oligosaccharides 455:506 rigorously characterized xyloglucan oligosaccharides 455:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 7 33 theme side-chain 1288:1297 arg1 structures 1299:1308 xyloglucan side-chain structures 1277:1308 xyloglucan side-chain structures 1277:1308 A comprehensive update of the nomenclature describing xyloglucan side-chain structures is provided for reference. 25497333 2 34 theme collection 441:450 arg1 availability 384:395 the availability 380:395 the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides 380:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 6 35 theme chemical 1145:1152 arg1 shifts 1154:1159 1H chemical shifts 1142:1159 1H chemical shifts 1142:1159 1H chemical shifts were assigned using two-dimensional correlation spectroscopy. 25497333 3 36 theme chemical 683:690 arg1 treatments 692:701 enzymatic and chemical treatments 669:701 enzymatic and chemical treatments 669:701 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 6 37 theme 1H 1142:1143 arg1 shifts 1154:1159 1H chemical shifts 1142:1159 1H chemical shifts 1142:1159 1H chemical shifts were assigned using two-dimensional correlation spectroscopy. 25497333 4 38 theme nuclear 981:987 arg1 resonance 998:1006 1H nuclear magnetic resonance 978:1006 1H nuclear magnetic resonance spectroscopy 978:1019 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 3 39 theme semi-preparative 545:560 arg1 generation 568:577 semi-preparative scale generation 545:577 semi-preparative scale generation 545:577 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 0 40 theme library 42:48 arg1 Generation 0:9 Generation 0:9 Generation 0:9 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 0 40 theme library 42:48 arg1 validation 26:35 structural validation 15:35 structural validation 15:35 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 7 41 theme xyloglucan 1277:1286 arg1 structures 1299:1308 xyloglucan side-chain structures 1277:1308 xyloglucan side-chain structures 1277:1308 A comprehensive update of the nomenclature describing xyloglucan side-chain structures is provided for reference. 25497333 3 42 theme enzymatic 669:677 arg1 treatments 692:701 enzymatic and chemical treatments 669:701 enzymatic and chemical treatments 669:701 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 0 43 link xyloglucan-derived 61:78 arg1 oligosaccharides 80:95 diverse xyloglucan-derived oligosaccharides 53:95 diverse xyloglucan-derived oligosaccharides 53:95 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 0 43 link xyloglucan-derived 61:78 arg1 update 111:116 an update 108:116 an update on xyloglucan nomenclature 108:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 3 44 theme scale 562:566 arg1 generation 568:577 semi-preparative scale generation 545:577 semi-preparative scale generation 545:577 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 4 45 theme exchange 949:956 arg1 chromatography 958:971 high-performance anion exchange chromatography 926:971 high-performance anion exchange chromatography 926:971 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 0 46 from update 111:116 arg1 nomenclature 132:143 xyloglucan nomenclature 121:143 xyloglucan nomenclature 121:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 3 47 theme size-exclusion 707:720 arg1 chromatography 722:735 size-exclusion chromatography 707:735 size-exclusion chromatography 707:735 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 7 48 theme comprehensive 1225:1237 arg1 update 1239:1244 A comprehensive update 1223:1244 A comprehensive update of the nomenclature describing xyloglucan side-chain structures 1223:1308 A comprehensive update of the nomenclature describing xyloglucan side-chain structures is provided for reference. 25497333 5 49 theme xyloglucan 1099:1108 arg1 quantification 1126:1139 xyloglucan oligosaccharide quantification 1099:1139 xyloglucan oligosaccharide quantification 1099:1139 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 4 50 theme laser 879:883 arg1 spectrometry 912:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 matrix-assisted laser desorption/ionization mass spectrometry 863:923 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 7 51 theme nomenclature 1253:1264 arg1 update 1239:1244 A comprehensive update 1223:1244 A comprehensive update of the nomenclature describing xyloglucan side-chain structures 1223:1308 A comprehensive update of the nomenclature describing xyloglucan side-chain structures is provided for reference. 25497333 1 52 dep cell 236:239 arg1 expansion 252:260 expansion 252:260 expansion 252:260 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 52 dep cell 236:239 arg1 metabolism 270:279 energy metabolism 263:279 energy metabolism 263:279 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 52 dep cell 236:239 arg1 signaling 286:294 signaling 286:294 signaling 286:294 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 1 52 dep cell 236:239 arg1 growth 241:246 growth 241:246 growth 241:246 Xyloglucans are structurally complex plant cell wall polysaccharides that are involved in cell growth and expansion, energy metabolism, and signaling. 25497333 5 53 theme oligosaccharide 1110:1124 arg1 quantification 1126:1139 xyloglucan oligosaccharide quantification 1099:1139 xyloglucan oligosaccharide quantification 1099:1139 Mass spectrometry and analytical chromatography were compared as methods for xyloglucan oligosaccharide quantification. 25497333 4 54 theme anion 943:947 arg1 chromatography 958:971 high-performance anion exchange chromatography 926:971 high-performance anion exchange chromatography 926:971 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 25497333 0 55 theme xyloglucan-derived 61:78 arg1 oligosaccharides 80:95 diverse xyloglucan-derived oligosaccharides 53:95 diverse xyloglucan-derived oligosaccharides 53:95 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 0 55 theme xyloglucan-derived 61:78 arg1 update 111:116 an update 108:116 an update on xyloglucan nomenclature 108:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 3 56 theme treatments 692:701 arg1 combination 654:664 a combination 652:664 a combination of enzymatic and chemical treatments and size-exclusion chromatography 652:735 Here, we present a workflow for the semi-preparative scale generation and purification of neutral and acidic xyloglucan oligosaccharides using a combination of enzymatic and chemical treatments and size-exclusion chromatography. 25497333 2 57 theme comprehensive 402:414 arg1 collection 441:450 a comprehensive and structurally diverse collection 400:450 a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides 400:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 2 58 theme diverse 433:439 arg1 collection 441:450 a comprehensive and structurally diverse collection 400:450 a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides 400:506 Determining the structure-function relationships of xyloglucans would benefit from the availability of a comprehensive and structurally diverse collection of rigorously characterized xyloglucan oligosaccharides. 25497333 0 59 theme diverse 53:59 arg1 oligosaccharides 80:95 diverse xyloglucan-derived oligosaccharides 53:95 diverse xyloglucan-derived oligosaccharides 53:95 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 0 59 theme diverse 53:59 arg1 update 111:116 an update 108:116 an update on xyloglucan nomenclature 108:143 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 0 60 theme structural 15:24 arg1 validation 26:35 structural validation 15:35 structural validation 15:35 Generation and structural validation of a library of diverse xyloglucan-derived oligosaccharides, including an update on xyloglucan nomenclature. 25497333 4 61 theme near 792:795 arg1 homogeneity 797:807 near homogeneity 792:807 near homogeneity 792:807 Twenty-six of these oligosaccharides were purified to near homogeneity and their structures validated using a combination of matrix-assisted laser desorption/ionization mass spectrometry, high-performance anion exchange chromatography, and 1H nuclear magnetic resonance spectroscopy. 26855252 3 0 theme hybrid 717:722 arg1 oligosaccharides 724:739 hybrid oligosaccharides 717:739 hybrid oligosaccharides 717:739 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 3 1 theme oligosaccharides 724:739 arg1 identification 699:712 The identification 695:712 The identification of hybrid oligosaccharides 695:739 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 3 2 theme starting 860:867 arg1 mixtures 869:876 the starting mixtures 856:876 the starting mixtures 856:876 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 4 3 theme oligosaccharides 941:956 arg1 proportions 922:932 different proportions 912:932 different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment 912:1014 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 1 4 theme different 186:194 arg1 proportions 202:212 different molar proportions 186:212 different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 186:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 4 5 from extents 987:993 arg1 proportions 922:932 different proportions 912:932 different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment 912:1014 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 0 6 theme Bean 105:108 arg1 Regions 110:116 Coffee Bean Regions 98:116 Coffee Bean Regions with Distinct Polysaccharide Composition 98:157 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 5 7 theme cell 1191:1194 arg1 walls 1196:1200 the bean cell walls 1182:1200 the bean cell walls 1182:1200 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 0 8 theme Coffee 98:103 arg1 Regions 110:116 Coffee Bean Regions 98:116 Coffee Bean Regions with Distinct Polysaccharide Composition 98:157 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 2 9 dep labeled 519:525 arg1 O 532:532 (18)O 528:532 (18)O 528:532 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 5 10 theme polysaccharides 1163:1177 arg1 distribution 1143:1154 the distribution 1139:1154 the distribution of the polysaccharides in the bean cell walls and the roasting conditions 1139:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 2 11 theme oligosaccharides 635:650 arg1 identification 565:578 identification 565:578 identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units 565:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 12 theme related 296:302 arg1 oligosaccharides 266:281 oligosaccharides 266:281 oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans) 266:366 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 1 12 theme related 296:302 arg1 mixtures 166:173 Three mixtures 160:173 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 160:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 2 13 theme labeled 519:525 arg1 samples 549:555 labeled ((18)O) and unlabeled samples 519:555 labeled ((18)O) and unlabeled samples 519:555 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 14 contain containing 175:184 arg1 mixtures 166:173 Three mixtures 160:173 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 160:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 1 14 contain containing 175:184 arg2 proportions 202:212 different molar proportions 186:212 different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 186:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 1 14 contain containing 175:184 arg1 oligosaccharides 266:281 oligosaccharides 266:281 oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans) 266:366 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 5 15 from walls 1196:1200 arg1 distribution 1143:1154 the distribution 1139:1154 the distribution of the polysaccharides in the bean cell walls and the roasting conditions 1139:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 2 16 theme hybrid 628:633 arg1 oligosaccharides 635:650 hybrid oligosaccharides 628:650 not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units 583:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 17 theme molar 196:200 arg1 proportions 202:212 different molar proportions 186:212 different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 186:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 4 18 from mixtures 974:981 arg1 proportions 922:932 different proportions 912:932 different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment 912:1014 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 2 19 theme spectrometry 444:455 arg1 analyses 507:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses 415:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples 415:555 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 20 theme pentose 680:686 arg1 units 688:692 both hexose and pentose units 664:692 units 688:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 21 theme tandem 470:475 arg1 ESI-MS 496:501 ESI-MS 496:501 ESI-MS(n) 496:504 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 21 theme tandem 470:475 arg1 spectrometry 482:493 tandem mass spectrometry 470:493 tandem mass spectrometry (ESI-MS(n)) 470:505 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 22 theme Electrospray 415:426 arg1 spectrometry 444:455 Electrospray ionization mass spectrometry 415:455 Electrospray ionization mass spectrometry (ESI-MS) 415:464 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 22 theme Electrospray 415:426 arg1 ESI-MS 458:463 ESI-MS 458:463 ESI-MS 458:463 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 0 23 theme Transglycosylation 13:30 arg1 Reactions 32:40 Nonenzymatic Transglycosylation Reactions 0:40 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 0:158 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 1 24 theme different 396:404 arg1 periods 406:412 different periods 396:412 different periods 396:412 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 2 25 theme oligosaccharides 602:617 arg1 identification 565:578 identification 565:578 identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units 565:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 0 26 with Regions 110:116 arg1 Composition 147:157 Distinct Polysaccharide Composition 123:157 Distinct Polysaccharide Composition 123:157 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 4 27 theme different 912:920 arg1 proportions 922:932 different proportions 912:932 different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment 912:1014 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 0 28 theme Nonenzymatic 0:11 arg1 Reactions 32:40 Nonenzymatic Transglycosylation Reactions 0:40 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 0:158 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 5 29 theme bean 1186:1189 arg1 walls 1196:1200 the bean cell walls 1182:1200 the bean cell walls 1182:1200 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 2 30 theme nonhybrid 592:600 arg1 oligosaccharides 602:617 nonhybrid oligosaccharides 592:617 not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units 583:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 31 theme hexose 669:674 arg1 units 688:692 both hexose and pentose units 664:692 units 688:692 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 32 dep -L-arabinotriose 248:263 arg1 α1→5 243:246 α1→5 243:246 α1→5 243:246 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 4 33 theme compounds 1061:1069 arg1 composition 1042:1052 the composition 1038:1052 the composition of the compounds formed 1038:1076 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 1 34 theme coffee 307:312 arg1 polysaccharides 314:328 coffee polysaccharides 307:328 coffee polysaccharides (galactomannans and arabinogalactans) 307:366 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 0 35 theme Polysaccharide 132:145 arg1 Composition 147:157 Distinct Polysaccharide Composition 123:157 Distinct Polysaccharide Composition 123:157 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 1 36 from °C 389:390 arg1 roasted 374:380 roasted 374:380 roasted 374:380 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 0 37 theme Distinct 123:130 arg1 Composition 147:157 Distinct Polysaccharide Composition 123:157 Distinct Polysaccharide Composition 123:157 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 5 38 from distribution 1143:1154 arg1 conditions 1219:1228 the roasting conditions 1206:1228 the roasting conditions 1206:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 5 38 from distribution 1143:1154 arg1 walls 1196:1200 the bean cell walls 1182:1200 the bean cell walls 1182:1200 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 4 39 theme starting 965:972 arg1 mixtures 974:981 the starting mixtures 961:981 the starting mixtures 961:981 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 0 40 from Models 81:86 arg1 Insights 67:74 New Insights 63:74 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 0:158 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 5 41 from conditions 1219:1228 arg1 distribution 1143:1154 the distribution 1139:1154 the distribution of the polysaccharides in the bean cell walls and the roasting conditions 1139:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 1 42 theme -D-mannotriose 223:236 arg1 proportions 202:212 different molar proportions 186:212 different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 186:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 2 43 theme ionization 428:437 arg1 spectrometry 444:455 Electrospray ionization mass spectrometry 415:455 Electrospray ionization mass spectrometry (ESI-MS) 415:464 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 43 theme ionization 428:437 arg1 ESI-MS 458:463 ESI-MS 458:463 ESI-MS 458:463 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 3 44 theme nonenzymatic 779:790 arg1 reactions 811:819 nonenzymatic transglycosylation reactions 779:819 nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures 779:876 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 5 45 theme roasting 1210:1217 arg1 conditions 1219:1228 the roasting conditions 1206:1228 the roasting conditions 1206:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 2 46 theme mass 439:442 arg1 spectrometry 444:455 Electrospray ionization mass spectrometry 415:455 Electrospray ionization mass spectrometry (ESI-MS) 415:464 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 46 theme mass 439:442 arg1 ESI-MS 458:463 ESI-MS 458:463 ESI-MS 458:463 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 0 47 theme New 63:65 arg1 Insights 67:74 New Insights 63:74 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 0:158 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 3 48 theme transglycosylation 792:809 arg1 reactions 811:819 nonenzymatic transglycosylation reactions 779:819 nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures 779:876 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 4 49 from oligosaccharides 941:956 arg1 extents 987:993 extents 987:993 extents 987:993 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 4 49 from oligosaccharides 941:956 arg1 mixtures 974:981 the starting mixtures 961:981 the starting mixtures 961:981 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 2 50 theme spectrometry 482:493 arg1 analyses 507:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses 415:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples 415:555 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 4 51 from variation 1025:1033 arg1 composition 1042:1052 the composition 1038:1052 the composition of the compounds formed 1038:1076 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 2 52 theme samples 549:555 arg1 analyses 507:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses 415:514 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples 415:555 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 53 theme unlabeled 539:547 arg1 samples 549:555 labeled ((18)O) and unlabeled samples 519:555 labeled ((18)O) and unlabeled samples 519:555 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 54 dep -D-mannotriose 223:236 arg1 β1→4 218:221 β1→4 218:221 β1→4 218:221 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 5 55 theme coffee 1294:1299 arg1 roasting 1301:1308 coffee roasting 1294:1308 coffee roasting 1294:1308 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 5 56 theme different 1231:1239 arg1 nonhybrid 1241:1249 different nonhybrid and hybrid structures 1231:1271 nonhybrid 1241:1249 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 3 57 from oligosaccharides 836:851 arg1 mixtures 869:876 the starting mixtures 856:876 the starting mixtures 856:876 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 4 58 from proportions 922:932 arg1 extents 987:993 extents 987:993 extents 987:993 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 4 58 from proportions 922:932 arg1 mixtures 974:981 the starting mixtures 961:981 the starting mixtures 961:981 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 0 59 dep Reactions 32:40 arg1 Insights 67:74 New Insights 63:74 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 0:158 Nonenzymatic Transglycosylation Reactions Induced by Roasting: New Insights from Models Mimicking Coffee Bean Regions with Distinct Polysaccharide Composition. 26855252 4 60 theme treatment 1006:1014 arg1 extents 987:993 extents 987:993 extents 987:993 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 4 60 theme treatment 1006:1014 arg1 mixtures 974:981 the starting mixtures 961:981 the starting mixtures 961:981 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 5 61 from polysaccharides 1163:1177 arg1 conditions 1219:1228 the roasting conditions 1206:1228 the roasting conditions 1206:1228 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 5 61 from polysaccharides 1163:1177 arg1 walls 1196:1200 the bean cell walls 1182:1200 the bean cell walls 1182:1200 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 3 62 theme reactions 811:819 arg1 occurrence 765:774 the occurrence 761:774 the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures 761:876 The identification of hybrid oligosaccharides allowed us to infer the occurrence of nonenzymatic transglycosylation reactions involving both oligosaccharides in the starting mixtures. 26855252 4 63 theme thermal 998:1004 arg1 treatment 1006:1014 thermal treatment 998:1014 thermal treatment 998:1014 Also, it was observed that using different proportions of the oligosaccharides in the starting mixtures and extents of thermal treatment led to a variation in the composition of the compounds formed. 26855252 1 64 dep polysaccharides 314:328 arg1 arabinogalactans 350:365 arabinogalactans 350:365 arabinogalactans 350:365 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 1 64 dep polysaccharides 314:328 arg1 galactomannans 331:344 galactomannans 331:344 galactomannans 331:344 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 26855252 5 65 theme hybrid 1255:1260 arg1 structures 1262:1271 different nonhybrid and hybrid structures 1231:1271 structures 1262:1271 These results have led to the conclusion that, depending on the distribution of the polysaccharides in the bean cell walls and the roasting conditions, different nonhybrid and hybrid structures can be formed during coffee roasting. 26855252 2 66 theme mass 477:480 arg1 ESI-MS 496:501 ESI-MS 496:501 ESI-MS(n) 496:504 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 2 66 theme mass 477:480 arg1 spectrometry 482:493 tandem mass spectrometry 470:493 tandem mass spectrometry (ESI-MS(n)) 470:505 Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS(n)) analyses of labeled ((18)O) and unlabeled samples allowed identification of not only nonhybrid oligosaccharides but also hybrid oligosaccharides composed of both hexose and pentose units. 26855252 1 67 theme -L-arabinotriose 248:263 arg1 proportions 202:212 different molar proportions 186:212 different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose 186:263 Three mixtures containing different molar proportions of (β1→4)-D-mannotriose and (α1→5)-L-arabinotriose, oligosaccharides structurally related to coffee polysaccharides (galactomannans and arabinogalactans), were roasted at 200 °C for different periods. 25115776 0 0 theme α-glucans 75:83 arg1 synthesis 42:50 synthesis 42:50 synthesis of linear and branched α-glucans 42:83 Hydrogen bond mediated aglycone delivery: synthesis of linear and branched α-glucans. 25115776 0 1 theme branched 66:73 arg1 α-glucans 75:83 linear and branched α-glucans 55:83 linear and branched α-glucans 55:83 Hydrogen bond mediated aglycone delivery: synthesis of linear and branched α-glucans. 25115776 2 2 theme sequences 313:321 arg1 synthesis 277:285 The synthesis 273:285 The synthesis of linear oligosaccharide sequences 273:321 The synthesis of linear oligosaccharide sequences was accomplished in complete stereoselectivity in all glycosylations. 25115776 2 3 theme oligosaccharide 297:311 arg1 sequences 313:321 linear oligosaccharide sequences 290:321 linear oligosaccharide sequences 290:321 The synthesis of linear oligosaccharide sequences was accomplished in complete stereoselectivity in all glycosylations. 25115776 4 4 theme structure 591:599 arg1 synthesis 567:575 The synthesis 563:575 The synthesis of a branched structure 563:599 The synthesis of a branched structure proved more challenging, particularly with bulky trisaccharide acceptors. 25115776 3 5 theme important 490:498 arg1 factor 500:505 an important factor 487:505 an important factor for the syntheses of oligomers beyond pentasaccharides 487:560 The efficacy of HAD may diminish with the increased bulk of the glycosyl acceptor, and may be an important factor for the syntheses of oligomers beyond pentasaccharides. 25115776 1 6 dep method 135:140 arg1 HAD 130:132 HAD 130:132 HAD 130:132 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 4 7 theme bulky 644:648 arg1 acceptors 664:672 bulky trisaccharide acceptors 644:672 bulky trisaccharide acceptors 644:672 The synthesis of a branched structure proved more challenging, particularly with bulky trisaccharide acceptors. 25115776 0 8 theme Hydrogen 0:7 arg1 bond 9:12 Hydrogen bond 0:12 Hydrogen bond 0:12 Hydrogen bond mediated aglycone delivery: synthesis of linear and branched α-glucans. 25115776 3 9 theme oligomers 528:536 arg1 syntheses 515:523 the syntheses 511:523 the syntheses of oligomers beyond pentasaccharides 511:560 The efficacy of HAD may diminish with the increased bulk of the glycosyl acceptor, and may be an important factor for the syntheses of oligomers beyond pentasaccharides. 25115776 2 10 theme complete 343:350 arg1 stereoselectivity 352:368 complete stereoselectivity 343:368 complete stereoselectivity in all glycosylations 343:390 The synthesis of linear oligosaccharide sequences was accomplished in complete stereoselectivity in all glycosylations. 25115776 0 11 theme aglycone 23:30 arg1 delivery 32:39 aglycone delivery 23:39 aglycone delivery 23:39 Hydrogen bond mediated aglycone delivery: synthesis of linear and branched α-glucans. 25115776 1 12 theme Hydrogen 88:95 arg1 bond 97:100 A Hydrogen bond 86:100 A Hydrogen bond mediated aglycone delivery (HAD) method 86:140 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 3 13 theme increased 435:443 arg1 bulk 445:448 the increased bulk 431:448 the increased bulk of the glycosyl acceptor 431:473 The efficacy of HAD may diminish with the increased bulk of the glycosyl acceptor, and may be an important factor for the syntheses of oligomers beyond pentasaccharides. 25115776 1 14 theme notable 242:248 arg1 challenge 250:258 a notable challenge 240:258 a notable challenge to chemists 240:270 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 3 15 theme acceptor 466:473 arg1 bulk 445:448 the increased bulk 431:448 the increased bulk of the glycosyl acceptor 431:473 The efficacy of HAD may diminish with the increased bulk of the glycosyl acceptor, and may be an important factor for the syntheses of oligomers beyond pentasaccharides. 25115776 1 16 theme α-glucans 174:182 arg1 synthesis 161:169 the synthesis 157:169 the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists 157:270 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 2 17 theme linear 290:295 arg1 sequences 313:321 linear oligosaccharide sequences 290:321 linear oligosaccharide sequences 290:321 The synthesis of linear oligosaccharide sequences was accomplished in complete stereoselectivity in all glycosylations. 25115776 2 18 from stereoselectivity 352:368 arg1 glycosylations 377:390 all glycosylations 373:390 all glycosylations 373:390 The synthesis of linear oligosaccharide sequences was accomplished in complete stereoselectivity in all glycosylations. 25115776 1 19 dep abundant 195:202 arg1 represent 230:238 represent 230:238 represent a notable challenge to chemists 230:270 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 3 20 theme glycosyl 457:464 arg1 acceptor 466:473 the glycosyl acceptor 453:473 the glycosyl acceptor 453:473 The efficacy of HAD may diminish with the increased bulk of the glycosyl acceptor, and may be an important factor for the syntheses of oligomers beyond pentasaccharides. 25115776 1 21 from nature 207:212 arg1 abundant 195:202 abundant 195:202 abundant 195:202 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 1 22 theme aglycone 111:118 arg1 method 135:140 aglycone delivery (HAD) method 111:140 aglycone delivery (HAD) method 111:140 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 1 23 theme delivery 120:127 arg1 method 135:140 aglycone delivery (HAD) method 111:140 aglycone delivery (HAD) method 111:140 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 1 24 from abundant 195:202 arg1 nature 207:212 nature 207:212 nature 207:212 A Hydrogen bond mediated aglycone delivery (HAD) method was applied to the synthesis of α-glucans, which are abundant in nature, but as targets represent a notable challenge to chemists. 25115776 4 25 theme branched 582:589 arg1 structure 591:599 a branched structure 580:599 a branched structure 580:599 The synthesis of a branched structure proved more challenging, particularly with bulky trisaccharide acceptors. 25115776 0 26 theme linear 55:60 arg1 α-glucans 75:83 linear and branched α-glucans 55:83 linear and branched α-glucans 55:83 Hydrogen bond mediated aglycone delivery: synthesis of linear and branched α-glucans. 25115776 4 27 theme trisaccharide 650:662 arg1 acceptors 664:672 bulky trisaccharide acceptors 644:672 bulky trisaccharide acceptors 644:672 The synthesis of a branched structure proved more challenging, particularly with bulky trisaccharide acceptors. 25101800 1 0 theme cell 125:128 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 0 theme cell 125:128 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 0 1 theme repeating 68:76 arg1 unit 78:81 cell wall trisaccharide repeating unit 44:81 cell wall trisaccharide repeating unit 44:81 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 1 2 theme wall 130:133 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 2 theme wall 130:133 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 3 3 theme linker 536:541 arg1 glycosylations 490:503 two consecutive α-stereoselective glycosylations 456:503 two consecutive α-stereoselective glycosylations 456:503 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 3 theme linker 536:541 arg1 attachment 518:527 β-selective attachment 506:527 β-selective attachment of the linker by solvent participation 506:566 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 4 theme solvent 546:552 arg1 participation 554:566 solvent participation 546:566 solvent participation 546:566 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 2 5 theme aminopropyl 239:249 arg1 linker 251:256 an aminopropyl linker 236:256 an aminopropyl linker 236:256 The synthetic trisaccharide is appended with an aminopropyl linker at the reducing end to allow for conjugation to proteins and microarrays. 25101800 1 6 theme trisaccharide 135:147 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 6 theme trisaccharide 135:147 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 3 7 theme d-mannose 384:392 arg1 steps 602:606 key steps 598:606 key steps 598:606 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 7 theme d-mannose 384:392 arg1 transformation 366:379 transformation 366:379 transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation 366:592 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 1 8 theme repeating 149:157 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 8 theme repeating 149:157 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 0 9 theme First 0:4 arg1 synthesis 6:14 First synthesis 0:14 First synthesis of Bacillus cereus Ch 0:36 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 0 10 theme Bacillus 19:26 arg1 Ch 35:36 Bacillus cereus Ch 19:36 Bacillus cereus Ch 19:36 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 2 11 theme reducing 265:272 arg1 end 274:276 the reducing end 261:276 the reducing end to allow for conjugation to proteins and microarrays 261:329 The synthetic trisaccharide is appended with an aminopropyl linker at the reducing end to allow for conjugation to proteins and microarrays. 25101800 3 12 theme building 440:447 arg1 block 449:453 an orthogonally protected rare AAT sugar building block 399:453 an orthogonally protected rare AAT sugar building block 399:453 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 13 theme convergent 336:345 arg1 synthesis 347:355 The convergent synthesis 332:355 The convergent synthesis 332:355 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 1 14 theme total 94:98 arg1 synthesis 100:108 The first total synthesis 84:108 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, 84:176 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 15 theme cereus 170:175 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 15 theme cereus 170:175 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 0 16 theme Ch 35:36 arg1 synthesis 6:14 First synthesis 0:14 First synthesis of Bacillus cereus Ch 0:36 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 2 17 theme synthetic 195:203 arg1 trisaccharide 205:217 The synthetic trisaccharide 191:217 The synthetic trisaccharide 191:217 The synthetic trisaccharide is appended with an aminopropyl linker at the reducing end to allow for conjugation to proteins and microarrays. 25101800 3 18 theme AAT 430:432 arg1 block 449:453 an orthogonally protected rare AAT sugar building block 399:453 an orthogonally protected rare AAT sugar building block 399:453 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 19 theme protected 415:423 arg1 block 449:453 an orthogonally protected rare AAT sugar building block 399:453 an orthogonally protected rare AAT sugar building block 399:453 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 20 theme sugar 434:438 arg1 block 449:453 an orthogonally protected rare AAT sugar building block 399:453 an orthogonally protected rare AAT sugar building block 399:453 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 0 21 theme cell 44:47 arg1 unit 78:81 cell wall trisaccharide repeating unit 44:81 cell wall trisaccharide repeating unit 44:81 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 3 22 theme amide 573:577 arg1 formation 584:592 amide bond formation 573:592 amide bond formation 573:592 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 23 theme consecutive 460:470 arg1 glycosylations 490:503 two consecutive α-stereoselective glycosylations 456:503 two consecutive α-stereoselective glycosylations 456:503 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 23 theme consecutive 460:470 arg1 attachment 518:527 β-selective attachment 506:527 β-selective attachment of the linker by solvent participation 506:566 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 24 theme bond 579:582 arg1 formation 584:592 amide bond formation 573:592 amide bond formation 573:592 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 25 theme α-stereoselective 472:488 arg1 glycosylations 490:503 two consecutive α-stereoselective glycosylations 456:503 two consecutive α-stereoselective glycosylations 456:503 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 25 theme α-stereoselective 472:488 arg1 attachment 518:527 β-selective attachment 506:527 β-selective attachment of the linker by solvent participation 506:566 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 0 26 dep Bacillus 19:26 arg1 cereus 28:33 cereus 28:33 cereus 28:33 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 1 27 theme Ch 113:114 arg1 unit 159:162 a cell wall trisaccharide repeating unit 123:162 a cell wall trisaccharide repeating unit of B. cereus 123:175 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 27 theme Ch 113:114 arg1 HF-PS 116:120 Ch HF-PS 113:120 Ch HF-PS 113:120 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 28 theme first 88:92 arg1 synthesis 100:108 The first total synthesis 84:108 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, 84:176 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 1 29 theme HF-PS 116:120 arg1 synthesis 100:108 The first total synthesis 84:108 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, 84:176 The first total synthesis of Ch HF-PS, a cell wall trisaccharide repeating unit of B. cereus, is reported. 25101800 0 30 theme trisaccharide 54:66 arg1 unit 78:81 cell wall trisaccharide repeating unit 44:81 cell wall trisaccharide repeating unit 44:81 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 3 31 theme β-selective 506:516 arg1 glycosylations 490:503 two consecutive α-stereoselective glycosylations 456:503 two consecutive α-stereoselective glycosylations 456:503 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 31 theme β-selective 506:516 arg1 attachment 518:527 β-selective attachment 506:527 β-selective attachment of the linker by solvent participation 506:566 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 32 theme rare 425:428 arg1 block 449:453 an orthogonally protected rare AAT sugar building block 399:453 an orthogonally protected rare AAT sugar building block 399:453 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 0 33 theme wall 49:52 arg1 unit 78:81 cell wall trisaccharide repeating unit 44:81 cell wall trisaccharide repeating unit 44:81 First synthesis of Bacillus cereus Ch HF-PS cell wall trisaccharide repeating unit. 25101800 3 34 theme key 598:600 arg1 steps 602:606 key steps 598:606 key steps 598:606 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25101800 3 34 theme key 598:600 arg1 transformation 366:379 transformation 366:379 transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation 366:592 The convergent synthesis involves transformation of d-mannose into an orthogonally protected rare AAT sugar building block, two consecutive α-stereoselective glycosylations, β-selective attachment of the linker by solvent participation, and amide bond formation, as key steps. 25725091 5 0 theme //bitbucket.org/fergaljd/glycoprofileassigner 797:841 arg1 CONTACT 843:849 //bitbucket.org/fergaljd/glycoprofileassigner CONTACT 797:849 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 0 1 with assignment 43:52 arg1 estimation 65:74 error estimation 59:74 error estimation 59:74 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 3 2 theme liquid 541:546 arg1 experiments 563:573 liquid chromatography experiments 541:573 liquid chromatography experiments 541:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 0 3 theme LC 87:88 arg1 data 90:93 glycan LC data 80:93 glycan LC data 80:93 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 1 4 dep MOTIVATION 96:105 arg1 structures 125:134 Sequencing glycan structures 107:134 MOTIVATION Sequencing glycan structures 96:134 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 2 5 theme sequential 310:319 arg1 digestions 336:345 sequential exoglycosidase digestions 310:345 sequential exoglycosidase digestions 310:345 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 1 6 theme glycan 118:123 arg1 structures 125:134 Sequencing glycan structures 107:134 MOTIVATION Sequencing glycan structures 96:134 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 5 7 dep INFORMATION 889:899 arg1 available 924:932 available 924:932 available 924:932 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 3 8 from experiments 563:573 arg1 data 531:534 glycan profile data 516:534 glycan profile data from liquid chromatography experiments 516:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 3 8 from experiments 563:573 arg1 assignment 502:511 automated structural assignment 481:511 automated structural assignment of glycan profile data from liquid chromatography experiments 481:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 3 9 theme profile 523:529 arg1 data 531:534 glycan profile data 516:534 glycan profile data from liquid chromatography experiments 516:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 5 10 dep AVAILABILITY 709:720 arg1 programme 745:753 The programme 741:753 The programme 741:753 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 2 11 theme chromatography 290:303 arg1 approach 231:238 One powerful approach 218:238 One powerful approach to glycan sequencing 218:259 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 2 11 theme chromatography 290:303 arg1 combination 268:278 the combination 264:278 the combination of liquid chromatography with sequential exoglycosidase digestions 264:345 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 5 12 theme nibrt.ie 866:873 arg1 INFORMATION 889:899 pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION 853:899 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 2 13 theme liquid 283:288 arg1 chromatography 290:303 liquid chromatography 283:303 liquid chromatography 283:303 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 5 14 theme SUPPLEMENTARY 875:887 arg1 INFORMATION 889:899 pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION 853:899 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 4 15 from structure 669:677 arg1 peaks 695:699 14 out of 16 peaks 682:699 14 out of 16 peaks tested 682:706 RESULTS GlycoProfileAssigner has been tested on human IgG data, and can retrieve the correct structure in 14 out of 16 peaks tested. 25725091 1 16 theme multiple 184:191 arg1 approaches 206:215 multiple experimental approaches 184:215 multiple experimental approaches 184:215 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 2 17 dep combination 268:278 arg1 interpreting 357:368 interpreting 357:368 interpreting this can be difficult and time-consuming 357:409 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 0 18 theme structural 32:41 arg1 assignment 43:52 automated structural assignment 22:52 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 0:94 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 1 19 theme Sequencing 107:116 arg1 structures 125:134 Sequencing glycan structures 107:134 MOTIVATION Sequencing glycan structures 96:134 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 1 20 theme approaches 206:215 arg1 use 177:179 the use 173:179 the use of multiple experimental approaches 173:215 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 0 21 theme automated 22:30 arg1 assignment 43:52 automated structural assignment 22:52 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 0:94 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 4 22 theme IgG 630:632 arg1 data 634:637 human IgG data 624:637 human IgG data 624:637 RESULTS GlycoProfileAssigner has been tested on human IgG data, and can retrieve the correct structure in 14 out of 16 peaks tested. 25725091 3 23 from assignment 502:511 arg1 experiments 563:573 liquid chromatography experiments 541:573 liquid chromatography experiments 541:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 2 24 theme exoglycosidase 321:334 arg1 digestions 336:345 sequential exoglycosidase digestions 310:345 sequential exoglycosidase digestions 310:345 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 5 25 theme Supplementary 901:913 arg1 data 915:918 Supplementary data 901:918 Supplementary data 901:918 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 4 26 theme human 624:628 arg1 data 634:637 human IgG data 624:637 human IgG data 624:637 RESULTS GlycoProfileAssigner has been tested on human IgG data, and can retrieve the correct structure in 14 out of 16 peaks tested. 25725091 5 27 dep https 791:795 arg1 INFORMATION 889:899 pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION 853:899 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 5 27 dep https 791:795 arg1 CONTACT 843:849 //bitbucket.org/fergaljd/glycoprofileassigner CONTACT 797:849 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 1 28 theme experimental 193:204 arg1 approaches 206:215 multiple experimental approaches 184:215 multiple experimental approaches 184:215 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 3 29 theme automated 481:489 arg1 assignment 502:511 automated structural assignment 481:511 automated structural assignment of glycan profile data from liquid chromatography experiments 481:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 5 30 theme pauline.rudd 853:864 arg1 INFORMATION 889:899 pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION 853:899 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 2 31 theme glycan 243:248 arg1 sequencing 250:259 glycan sequencing 243:259 glycan sequencing 243:259 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 3 32 theme structural 491:500 arg1 assignment 502:511 automated structural assignment 481:511 automated structural assignment of glycan profile data from liquid chromatography experiments 481:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 5 33 theme @ 865:865 arg1 INFORMATION 889:899 pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION 853:899 https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online 791:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 4 34 theme RESULTS 576:582 arg1 GlycoProfileAssigner 584:603 RESULTS GlycoProfileAssigner 576:603 RESULTS GlycoProfileAssigner 576:603 RESULTS GlycoProfileAssigner has been tested on human IgG data, and can retrieve the correct structure in 14 out of 16 peaks tested. 25725091 3 35 theme data 531:534 arg1 assignment 502:511 automated structural assignment 481:511 automated structural assignment of glycan profile data from liquid chromatography experiments 481:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 0 36 theme error 59:63 arg1 estimation 65:74 error estimation 59:74 error estimation 59:74 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 2 37 theme powerful 222:229 arg1 approach 231:238 One powerful approach 218:238 One powerful approach to glycan sequencing 218:259 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 2 37 theme powerful 222:229 arg1 combination 268:278 the combination 264:278 the combination of liquid chromatography with sequential exoglycosidase digestions 264:345 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 3 38 theme chromatography 548:561 arg1 experiments 563:573 liquid chromatography experiments 541:573 liquid chromatography experiments 541:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 2 39 with combination 268:278 arg1 digestions 336:345 sequential exoglycosidase digestions 310:345 sequential exoglycosidase digestions 310:345 One powerful approach to glycan sequencing is the combination of liquid chromatography with sequential exoglycosidase digestions; however, interpreting this can be difficult and time-consuming. 25725091 3 40 theme glycan 516:521 arg1 data 531:534 glycan profile data 516:534 glycan profile data from liquid chromatography experiments 516:573 To aid this process, we introduce GlycoProfileAssigner, software for automated structural assignment of glycan profile data from liquid chromatography experiments. 25725091 5 41 from https 791:795 arg1 available 778:786 available 778:786 available 778:786 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 1 42 theme difficult 141:149 arg1 MOTIVATION 96:105 MOTIVATION Sequencing glycan structures 96:134 MOTIVATION Sequencing glycan structures 96:134 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 1 42 theme difficult 141:149 arg1 problem 151:157 a difficult problem 139:157 a difficult problem that requires the use of multiple experimental approaches 139:215 MOTIVATION Sequencing glycan structures is a difficult problem that requires the use of multiple experimental approaches. 25725091 0 43 theme glycan 80:85 arg1 data 90:93 glycan LC data 80:93 glycan LC data 80:93 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 5 44 theme Bioinformatics 937:950 arg1 online 952:957 Bioinformatics online 937:957 Bioinformatics online 937:957 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 0 45 dep GlycoProfileAssigner 0:19 arg1 assignment 43:52 automated structural assignment 22:52 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 0:94 GlycoProfileAssigner: automated structural assignment with error estimation for glycan LC data. 25725091 5 46 from online 952:957 arg1 available 924:932 available 924:932 available 924:932 AVAILABILITY AND IMPLEMENTATION The programme and its source code is available at https://bitbucket.org/fergaljd/glycoprofileassigner CONTACT : pauline.rudd@nibrt.ie SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 25725091 4 47 theme correct 661:667 arg1 structure 669:677 the correct structure 657:677 the correct structure in 14 out of 16 peaks tested 657:706 RESULTS GlycoProfileAssigner has been tested on human IgG data, and can retrieve the correct structure in 14 out of 16 peaks tested. 24873750 2 0 theme modulating 557:566 arg1 activity 568:575 intestinal immune system modulating activity 532:575 intestinal immune system modulating activity 532:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 1 1 theme efficient 157:165 arg1 synthesis 190:198 the efficient four-component one-pot synthesis 153:198 the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure 153:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 2 2 theme system 550:555 arg1 activity 568:575 intestinal immune system modulating activity 532:575 intestinal immune system modulating activity 532:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 1 3 theme four-component 167:180 arg1 synthesis 190:198 the efficient four-component one-pot synthesis 153:198 the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure 153:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 4 theme D-thiogalactopyranoside 279:301 arg1 3,6-diol 303:310 D-thiogalactopyranoside 3,6-diol 3 279:312 D-thiogalactopyranoside 3,6-diol 3 279:312 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 4 theme D-thiogalactopyranoside 279:301 arg1 agent 343:347 the central glycosylating agent 317:347 the central glycosylating agent 317:347 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 0 5 theme glycosylating 107:119 arg1 agent 121:125 a key glycosylating agent 101:125 a key glycosylating agent 101:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 5 theme glycosylating 107:119 arg1 diol 93:96 galactopyranosyl thioglycoside diol 62:96 galactopyranosyl thioglycoside diol 62:96 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 6 theme key 103:105 arg1 agent 121:125 a key glycosylating agent 101:125 a key glycosylating agent 101:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 6 theme key 103:105 arg1 diol 93:96 galactopyranosyl thioglycoside diol 62:96 galactopyranosyl thioglycoside diol 62:96 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 1 7 theme one-pot 182:188 arg1 synthesis 190:198 the efficient four-component one-pot synthesis 153:198 the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure 153:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 2 8 theme immune 543:548 arg1 system 550:555 intestinal immune system 532:555 intestinal immune system modulating activity 532:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 9 theme global 356:361 arg1 deprotection 363:374 global deprotection 356:374 global deprotection 356:374 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 10 theme free 434:437 arg1 oligosaccharide 439:453 the 3-aminopropyl linker-containing free oligosaccharide 1 398:455 the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity 398:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 10 theme free 434:437 arg1 related 478:484 related 478:484 related 478:484 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 0 11 theme One-pot 0:6 arg1 synthesis 8:16 One-pot synthesis 0:16 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 0:126 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 12 gly glycosylating 107:119 arg0 diol 93:96 galactopyranosyl thioglycoside diol 62:96 galactopyranosyl thioglycoside diol 62:96 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 12 gly glycosylating 107:119 arg0 agent 121:125 a key glycosylating agent 101:125 a key glycosylating agent 101:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 2 13 theme linker-containing 416:432 arg1 oligosaccharide 439:453 the 3-aminopropyl linker-containing free oligosaccharide 1 398:455 the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity 398:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 13 theme linker-containing 416:432 arg1 related 478:484 related 478:484 related 478:484 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 1 14 with hexaarabinogalactan 221:239 arg1 structure 260:268 di-branched structure 248:268 di-branched structure 248:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 15 theme central 321:327 arg1 3,6-diol 303:310 D-thiogalactopyranoside 3,6-diol 3 279:312 D-thiogalactopyranoside 3,6-diol 3 279:312 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 15 theme central 321:327 arg1 agent 343:347 the central glycosylating agent 317:347 the central glycosylating agent 317:347 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 16 theme protected 211:219 arg1 hexaarabinogalactan 221:239 a fully protected hexaarabinogalactan 2 203:241 a fully protected hexaarabinogalactan 2 with di-branched structure 203:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 0 17 theme hexaarabinogalactan 36:54 arg1 synthesis 8:16 One-pot synthesis 0:16 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 0:126 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 1 18 theme glycosylating 329:341 arg1 3,6-diol 303:310 D-thiogalactopyranoside 3,6-diol 3 279:312 D-thiogalactopyranoside 3,6-diol 3 279:312 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 18 theme glycosylating 329:341 arg1 agent 343:347 the central glycosylating agent 317:347 the central glycosylating agent 317:347 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 19 theme hexaarabinogalactan 221:239 arg1 synthesis 190:198 the efficient four-component one-pot synthesis 153:198 the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure 153:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 0 20 theme 3,6-branched 23:34 arg1 hexaarabinogalactan 36:54 a 3,6-branched hexaarabinogalactan 21:54 a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent 21:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 2 21 theme intestinal 532:541 arg1 system 550:555 intestinal immune system 532:555 intestinal immune system modulating activity 532:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 0 22 theme galactopyranosyl 62:77 arg1 agent 121:125 a key glycosylating agent 101:125 a key glycosylating agent 101:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 22 theme galactopyranosyl 62:77 arg1 diol 93:96 galactopyranosyl thioglycoside diol 62:96 galactopyranosyl thioglycoside diol 62:96 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 1 23 gly glycosylating 329:341 arg0 3,6-diol 303:310 D-thiogalactopyranoside 3,6-diol 3 279:312 D-thiogalactopyranoside 3,6-diol 3 279:312 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 23 gly glycosylating 329:341 arg0 agent 343:347 the central glycosylating agent 317:347 the central glycosylating agent 317:347 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 24 theme di-branched 248:258 arg1 structure 260:268 di-branched structure 248:268 di-branched structure 248:268 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 2 25 with arabino-3,6-galactan 506:525 arg1 activity 568:575 intestinal immune system modulating activity 532:575 intestinal immune system modulating activity 532:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 26 theme 3-aminopropyl 402:414 arg1 oligosaccharide 439:453 the 3-aminopropyl linker-containing free oligosaccharide 1 398:455 the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity 398:575 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 2 26 theme 3-aminopropyl 402:414 arg1 related 478:484 related 478:484 related 478:484 After global deprotection, 2 was converted into the 3-aminopropyl linker-containing free oligosaccharide 1 that is structurally related to ALR-5IIa-1-1, an arabino-3,6-galactan with intestinal immune system modulating activity. 24873750 0 27 theme thioglycoside 79:91 arg1 agent 121:125 a key glycosylating agent 101:125 a key glycosylating agent 101:125 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 0 27 theme thioglycoside 79:91 arg1 diol 93:96 galactopyranosyl thioglycoside diol 62:96 galactopyranosyl thioglycoside diol 62:96 One-pot synthesis of a 3,6-branched hexaarabinogalactan using galactopyranosyl thioglycoside diol as a key glycosylating agent. 24873750 1 28 attach present 131:137 arg2 We 128:129 We 128:129 We 128:129 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 24873750 1 28 attach present 131:137 arg1 paper 147:151 this paper 142:151 this paper 142:151 We present in this paper the efficient four-component one-pot synthesis of a fully protected hexaarabinogalactan 2 with di-branched structure by using D-thiogalactopyranoside 3,6-diol 3 as the central glycosylating agent. 26297472 0 0 theme twitching 67:75 arg1 motility 77:84 twitching motility 67:84 twitching motility 67:84 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 4 1 theme Culture 464:470 arg1 fluids 484:489 Culture supernatant fluids 464:489 Culture supernatant fluids containing fully, partially 464:517 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 7 2 theme Reduced 951:957 arg1 solubility 959:968 Reduced solubility 951:968 Reduced solubility 951:968 Reduced solubility suggested an increased pilus surface hydrophobicity, which was supported by protein modelling. 26297472 8 3 from effect 1081:1086 arg1 strain 1098:1103 the WT strain 1091:1103 the WT strain 1091:1103 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 6 4 theme group 868:872 arg1 pili 877:880 the P. aeruginosa group II pili 850:880 the P. aeruginosa group II pili 850:880 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 11 5 theme optimal 1593:1599 arg1 pilus 1601:1605 optimal pilus 1593:1605 optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here 1593:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 6 6 theme pili 877:880 arg1 representative 832:845 A representative 830:845 A representative of the P. aeruginosa group II pili 830:880 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 10 7 theme II 1400:1401 arg1 pili 1403:1406 group II pili 1394:1406 group II pili 1394:1406 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 4 8 theme I 560:560 arg1 pili 562:565 non-glycosylated P. aeruginosa group I pili 523:565 non-glycosylated P. aeruginosa group I pili 523:565 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 6 9 theme II 874:875 arg1 pili 877:880 the P. aeruginosa group II pili 850:880 the P. aeruginosa group II pili 850:880 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 10 10 theme strain 1365:1370 arg1 PA103 1372:1376 strain PA103 1365:1376 P. aeruginosa strain PA103 1351:1376 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 0 11 from IVa 38:40 arg1 1244 112:115 1244 112:115 1244 112:115 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 7 12 theme pilus 993:997 arg1 hydrophobicity 1007:1020 an increased pilus surface hydrophobicity 980:1020 an increased pilus surface hydrophobicity 980:1020 Reduced solubility suggested an increased pilus surface hydrophobicity, which was supported by protein modelling. 26297472 10 13 theme aeruginosa 1354:1363 arg1 motility 1339:1346 Twitching motility 1329:1346 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, 1329:1407 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 2 14 theme pili 253:256 arg1 surfaces 235:242 The lateral surfaces 223:242 The lateral surfaces of these pili 223:256 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 8 15 theme infection 1143:1151 arg1 sites 1153:1157 many host infection sites 1133:1157 many host infection sites 1133:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 16 theme many 1133:1136 arg1 sites 1153:1157 many host infection sites 1133:1157 many host infection sites 1133:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 7 17 theme surface 999:1005 arg1 hydrophobicity 1007:1020 an increased pilus surface hydrophobicity 980:1020 an increased pilus surface hydrophobicity 980:1020 Reduced solubility suggested an increased pilus surface hydrophobicity, which was supported by protein modelling. 26297472 5 18 theme salt 802:805 arg1 concentrations 807:820 the lowest salt concentrations 791:820 the lowest salt concentrations tested 791:827 These results showed that while pili expressing three or four sugars were highly soluble under all conditions, those with fewer than three were insoluble under the lowest salt concentrations tested. 26297472 3 19 theme pilus 433:437 arg1 solubility 439:448 pilus solubility 433:448 pilus solubility 433:448 The current work was conducted to investigate the pilin glycan's effect on pilus solubility and function. 26297472 8 20 theme 1244G7 1221:1226 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 20 theme 1244G7 1221:1226 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 3 21 theme pilin 408:412 arg1 glycan 414:419 pilin glycan's 408:421 the pilin glycan's effect on pilus solubility and function 404:461 The current work was conducted to investigate the pilin glycan's effect on pilus solubility and function. 26297472 8 22 theme subsurface 1181:1190 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 22 theme subsurface 1181:1190 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 4 23 dep P. 540:541 arg1 aeruginosa 543:552 aeruginosa 543:552 aeruginosa 543:552 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 10 24 theme group 1394:1398 arg1 pili 1403:1406 group II pili 1394:1406 group II pili 1394:1406 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 8 25 theme host 1138:1141 arg1 sites 1153:1157 many host infection sites 1133:1157 many host infection sites 1133:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 11 26 theme host 1631:1634 arg1 sites 1644:1648 the many host disease sites 1622:1648 the many host disease sites with ionic strengths comparable to those tested here 1622:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 0 27 theme group 4:8 arg1 glycan 18:23 The group I pilin glycan 0:23 The group I pilin glycan 0:23 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 4 28 theme ammonium 613:620 arg1 sulfate 622:628 ammonium sulfate 613:628 ammonium sulfate 613:628 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 1 29 theme common 157:162 arg1 type 164:167 the most common type 148:167 the most common type of type IVa pilus produced by Pseudomonas aeruginosa 148:220 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 1 29 theme common 157:162 arg1 category 136:143 The group I pilin category 118:143 The group I pilin category 118:143 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 3 30 theme current 362:368 arg1 work 370:373 The current work 358:373 The current work 358:373 The current work was conducted to investigate the pilin glycan's effect on pilus solubility and function. 26297472 4 31 theme P. 540:541 arg1 pili 562:565 non-glycosylated P. aeruginosa group I pili 523:565 non-glycosylated P. aeruginosa group I pili 523:565 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 11 32 theme comparable 1671:1680 arg1 strengths 1661:1669 ionic strengths 1655:1669 ionic strengths comparable to those tested here 1655:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 11 33 theme group 1508:1512 arg1 beneficial 1548:1557 beneficial 1548:1557 beneficial 1548:1557 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 11 33 theme group 1508:1512 arg1 glycan 1522:1527 the group I pilin glycan 1504:1527 the group I pilin glycan 1504:1527 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 8 34 located found 1124:1128 arg1 sites 1153:1157 many host infection sites 1133:1157 many host infection sites 1133:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 34 located found 1124:1128 arg2 strength 1115:1122 an ionic strength 1106:1122 an ionic strength found at many host infection sites 1106:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 35 theme ionic 1109:1113 arg1 strength 1115:1122 an ionic strength 1106:1122 an ionic strength found at many host infection sites 1106:1157 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 2 36 theme units 351:355 arg1 presence 283:290 the presence 279:290 the presence of closely spaced, covalently attached O-antigen repeating units 279:355 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 0 37 theme pilin 12:16 arg1 glycan 18:23 The group I pilin glycan 0:23 The group I pilin glycan 0:23 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 4 38 theme non-glycosylated 523:538 arg1 pili 562:565 non-glycosylated P. aeruginosa group I pili 523:565 non-glycosylated P. aeruginosa group I pili 523:565 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 7 39 theme protein 1046:1052 arg1 modelling 1054:1062 protein modelling 1046:1062 protein modelling 1046:1062 Reduced solubility suggested an increased pilus surface hydrophobicity, which was supported by protein modelling. 26297472 11 40 theme I 1514:1514 arg1 beneficial 1548:1557 beneficial 1548:1557 beneficial 1548:1557 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 11 40 theme I 1514:1514 arg1 glycan 1522:1527 the group I pilin glycan 1504:1527 the group I pilin glycan 1504:1527 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 9 41 theme mutant 1305:1310 arg1 complementation 1312:1326 mutant complementation 1305:1326 mutant complementation 1305:1326 This effect was reversed by mutant complementation. 26297472 5 42 theme lowest 795:800 arg1 concentrations 807:820 the lowest salt concentrations 791:820 the lowest salt concentrations tested 791:827 These results showed that while pili expressing three or four sugars were highly soluble under all conditions, those with fewer than three were insoluble under the lowest salt concentrations tested. 26297472 0 43 theme I 10:10 arg1 glycan 18:23 The group I pilin glycan 0:23 The group I pilin glycan 0:23 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 4 44 theme group 554:558 arg1 pili 562:565 non-glycosylated P. aeruginosa group I pili 523:565 non-glycosylated P. aeruginosa group I pili 523:565 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 2 45 theme repeating 341:349 arg1 units 351:355 closely spaced, covalently attached O-antigen repeating units 295:355 closely spaced, covalently attached O-antigen repeating units 295:355 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 11 46 theme disease 1636:1642 arg1 sites 1644:1648 the many host disease sites 1622:1648 the many host disease sites with ionic strengths comparable to those tested here 1622:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 2 47 dep spaced 303:308 arg1 attached 322:329 attached 322:329 attached 322:329 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 1 48 theme type 172:175 arg1 pilus 181:185 type IVa pilus 172:185 type IVa pilus 172:185 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 6 49 theme P. 854:855 arg1 pili 877:880 the P. aeruginosa group II pili 850:880 the P. aeruginosa group II pili 850:880 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 4 50 theme sulfate 622:628 arg1 presence 601:608 the presence 597:608 the presence of ammonium sulfate 597:628 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 0 51 dep IVa 38:40 arg1 hydrophobicity 48:61 pilus hydrophobicity 42:61 pilus hydrophobicity 42:61 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 8 52 theme strain 1214:1219 arg1 1244G7 1221:1226 strain 1244G7 1214:1226 strain 1244G7 1214:1226 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 1 53 theme IVa 177:179 arg1 pilus 181:185 type IVa pilus 172:185 type IVa pilus 172:185 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 8 54 theme surface 1169:1175 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 54 theme surface 1169:1175 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 11 55 theme ionic 1655:1659 arg1 strengths 1661:1669 ionic strengths 1655:1669 ionic strengths comparable to those tested here 1655:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 11 56 theme many 1626:1629 arg1 sites 1644:1648 the many host disease sites 1622:1648 the many host disease sites with ionic strengths comparable to those tested here 1622:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 10 57 dep mutant 1468:1473 arg1 strain 1480:1485 1244 strain 1475:1485 the mutant 1244 strain 1464:1485 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 4 58 gly non-glycosylated 523:538 arg1 pili 562:565 non-glycosylated P. aeruginosa group I pili 523:565 non-glycosylated P. aeruginosa group I pili 523:565 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 1 59 theme pilus 181:185 arg1 type 164:167 the most common type 148:167 the most common type of type IVa pilus produced by Pseudomonas aeruginosa 148:220 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 1 59 theme pilus 181:185 arg1 category 136:143 The group I pilin category 118:143 The group I pilin category 118:143 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 8 60 theme twitching 1192:1200 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 60 theme twitching 1192:1200 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 6 61 theme same 934:937 arg1 conditions 939:948 these same conditions 928:948 these same conditions 928:948 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 3 62 from effect 423:428 arg1 function 454:461 function 454:461 function 454:461 The current work was conducted to investigate the pilin glycan's effect on pilus solubility and function. 26297472 3 62 from effect 423:428 arg1 solubility 439:448 pilus solubility 433:448 pilus solubility 433:448 The current work was conducted to investigate the pilin glycan's effect on pilus solubility and function. 26297472 0 63 theme type 33:36 arg1 IVa 38:40 type IVa pilus hydrophobicity and twitching motility 33:84 IVa 38:40 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 1 64 theme pilin 130:134 arg1 type 164:167 the most common type 148:167 the most common type of type IVa pilus produced by Pseudomonas aeruginosa 148:220 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 1 64 theme pilin 130:134 arg1 category 136:143 The group I pilin category 118:143 The group I pilin category 118:143 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 2 65 theme spaced 303:308 arg1 units 351:355 closely spaced, covalently attached O-antigen repeating units 295:355 closely spaced, covalently attached O-antigen repeating units 295:355 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 6 66 theme low 894:896 arg1 solubility 898:907 low solubility 894:907 low solubility 894:907 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 8 67 theme WT 1095:1096 arg1 strain 1098:1103 the WT strain 1091:1103 the WT strain 1091:1103 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 11 68 theme pilin 1516:1520 arg1 beneficial 1548:1557 beneficial 1548:1557 beneficial 1548:1557 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 11 68 theme pilin 1516:1520 arg1 glycan 1522:1527 the group I pilin glycan 1504:1527 the group I pilin glycan 1504:1527 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 7 69 theme increased 983:991 arg1 hydrophobicity 1007:1020 an increased pilus surface hydrophobicity 980:1020 an increased pilus surface hydrophobicity 980:1020 Reduced solubility suggested an increased pilus surface hydrophobicity, which was supported by protein modelling. 26297472 8 70 gly glycosylate 1258:1268 arg1 pilin 1270:1274 pilin 1270:1274 pilin 1270:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 6 71 dep P. 854:855 arg1 aeruginosa 857:866 aeruginosa 857:866 aeruginosa 857:866 A representative of the P. aeruginosa group II pili also showed low solubility when assayed under these same conditions. 26297472 2 72 theme lateral 227:233 arg1 surfaces 235:242 The lateral surfaces 223:242 The lateral surfaces of these pili 223:256 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 10 73 theme ionic 1431:1435 arg1 strengths 1437:1445 ionic strengths 1431:1445 ionic strengths which influenced the mutant 1244 strain 1431:1485 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 1 74 theme group 122:126 arg1 type 164:167 the most common type 148:167 the most common type of type IVa pilus produced by Pseudomonas aeruginosa 148:220 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 1 74 theme group 122:126 arg1 category 136:143 The group I pilin category 118:143 The group I pilin category 118:143 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 11 75 with sites 1644:1648 arg1 strengths 1661:1669 ionic strengths 1655:1669 ionic strengths comparable to those tested here 1655:1701 We suggest that the group I pilin glycan may, therefore, be beneficial to this organism specifically for optimal pilus functioning at the many host disease sites with ionic strengths comparable to those tested here. 26297472 0 76 from motility 77:84 arg1 1244 112:115 1244 112:115 1244 112:115 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 8 77 theme isogenic 1232:1239 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 77 theme isogenic 1232:1239 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 10 78 dep aeruginosa 1354:1363 arg1 PA103 1372:1376 strain PA103 1365:1376 P. aeruginosa strain PA103 1351:1376 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 1 79 theme I 128:128 arg1 type 164:167 the most common type 148:167 the most common type of type IVa pilus produced by Pseudomonas aeruginosa 148:220 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 1 79 theme I 128:128 arg1 category 136:143 The group I pilin category 118:143 The group I pilin category 118:143 The group I pilin category is the most common type of type IVa pilus produced by Pseudomonas aeruginosa. 26297472 0 80 theme pilus 42:46 arg1 hydrophobicity 48:61 pilus hydrophobicity 42:61 pilus hydrophobicity 42:61 The group I pilin glycan affects type IVa pilus hydrophobicity and twitching motility in Pseudomonas aeruginosa 1244. 26297472 4 81 theme supernatant 472:482 arg1 fluids 484:489 Culture supernatant fluids 464:489 Culture supernatant fluids containing fully, partially 464:517 Culture supernatant fluids containing fully, partially and non-glycosylated P. aeruginosa group I pili were tested for solubility in the presence of ammonium sulfate. 26297472 10 82 theme Twitching 1329:1337 arg1 motility 1339:1346 Twitching motility 1329:1346 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, 1329:1407 Twitching motility of P. aeruginosa strain PA103, which produces group II pili, was also inhibited by ionic strengths which influenced the mutant 1244 strain. 26297472 2 83 theme O-antigen 331:339 arg1 units 351:355 closely spaced, covalently attached O-antigen repeating units 295:355 closely spaced, covalently attached O-antigen repeating units 295:355 The lateral surfaces of these pili are characterized by the presence of closely spaced, covalently attached O-antigen repeating units. 26297472 8 84 theme unable 1248:1253 arg1 motility 1202:1209 surface and subsurface twitching motility 1169:1209 motility 1202:1209 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26297472 8 84 theme unable 1248:1253 arg1 mutant 1241:1246 an isogenic mutant 1229:1246 an isogenic mutant unable to glycosylate pilin 1229:1274 While having no effect on the WT strain, an ionic strength found at many host infection sites inhibited surface and subsurface twitching motility of strain 1244G7, an isogenic mutant unable to glycosylate pilin. 26244886 7 0 theme published 1252:1260 arg1 denaturation 1262:1273 a published denaturation 1250:1273 a published denaturation 1250:1273 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 6 1 theme denatured 1051:1059 arg1 antibodies 1075:1084 denatured versus intact antibodies 1051:1084 denatured versus intact antibodies 1051:1084 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 2 2 theme proteomics 362:371 arg1 workflows 373:381 many proteomics workflows 357:381 many proteomics workflows 357:381 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 6 3 theme intact 957:962 arg1 structure 977:985 the intact higher order structure 953:985 the intact higher order structure of the antibodies 953:1003 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 2 4 theme many 357:360 arg1 workflows 373:381 many proteomics workflows 357:381 many proteomics workflows 357:381 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 5 5 theme alpha2-3-sialylated 778:796 arg1 mannose 761:767 high mannose 756:767 high mannose 756:767 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 5 theme alpha2-3-sialylated 778:796 arg1 glycoforms 811:820 alpha2-3-sialylated and bisected glycoforms 778:820 alpha2-3-sialylated and bisected glycoforms 778:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 4 6 theme glycan 687:692 arg1 compositions 694:705 individual glycan compositions 676:705 individual glycan compositions 676:705 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 4 7 theme biases 653:658 arg1 resolution 639:648 resolution 639:648 resolution of biases 639:658 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 7 8 theme comparable 1228:1237 arg1 results 1239:1245 highly comparable results 1221:1245 highly comparable results 1221:1245 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 2 9 from essential 344:352 arg1 workflows 373:381 many proteomics workflows 357:381 many proteomics workflows 357:381 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 0 10 theme Glycoform 105:113 arg1 Influence 115:123 the Glycoform Influence 101:123 the Glycoform Influence on Higher-Order Structure 101:149 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 5 11 dep neutral 858:864 arg1 fucosylated 867:877 fucosylated 867:877 fucosylated 867:877 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 6 12 theme higher 964:969 arg1 structure 977:985 the intact higher order structure 953:985 the intact higher order structure of the antibodies 953:1003 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 3 13 theme polyclonal 399:408 arg1 substances 469:478 model substances 463:478 model substances 463:478 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 13 theme polyclonal 399:408 arg1 biopharmaceuticals 427:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 6 14 theme large 926:930 arg1 extent 932:937 a large extent 924:937 a large extent 924:937 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 5 15 gly alpha2-3-sialylated 778:796 arg1 mannose 761:767 high mannose 756:767 high mannose 756:767 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 15 gly alpha2-3-sialylated 778:796 arg1 glycoforms 811:820 alpha2-3-sialylated and bisected glycoforms 778:820 alpha2-3-sialylated and bisected glycoforms 778:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 3 16 theme immunoglobulin 410:423 arg1 substances 469:478 model substances 463:478 model substances 463:478 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 16 theme immunoglobulin 410:423 arg1 biopharmaceuticals 427:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 4 17 theme Fc-glycosylation 609:624 arg1 site 626:629 the conserved Fc-glycosylation site 595:629 the conserved Fc-glycosylation site 595:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 7 18 theme acidic 1125:1130 arg1 denaturation 1132:1143 acidic denaturation 1125:1143 acidic denaturation 1125:1143 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 4 19 attach derived 582:588 arg2 glycopeptides 568:580 glycopeptides 568:580 glycopeptides derived from the conserved Fc-glycosylation site 568:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 4 19 attach derived 582:588 arg1 site 626:629 the conserved Fc-glycosylation site 595:629 the conserved Fc-glycosylation site 595:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 7 20 theme cleavage 1102:1109 arg1 protocol 1111:1118 a cleavage protocol 1100:1118 a cleavage protocol with acidic denaturation 1100:1143 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 3 21 theme model 463:467 arg1 substances 469:478 model substances 463:478 model substances 463:478 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 21 theme model 463:467 arg1 biopharmaceuticals 427:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 22 theme Monoclonal 384:393 arg1 substances 469:478 model substances 463:478 model substances 463:478 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 22 theme Monoclonal 384:393 arg1 biopharmaceuticals 427:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 4 23 theme conserved 599:607 arg1 site 626:629 the conserved Fc-glycosylation site 595:629 the conserved Fc-glycosylation site 595:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 0 24 theme Higher-Order 128:139 arg1 Structure 141:149 Higher-Order Structure 128:149 Higher-Order Structure 128:149 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 2 25 from workflows 373:381 arg1 essential 344:352 essential 344:352 essential 344:352 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 2 25 from workflows 373:381 arg1 trypsin 326:332 the serine protease trypsin 306:332 the serine protease trypsin 306:332 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 2 26 theme protease 317:324 arg1 essential 344:352 essential 344:352 essential 344:352 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 2 26 theme protease 317:324 arg1 trypsin 326:332 the serine protease trypsin 306:332 the serine protease trypsin 306:332 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 4 27 theme compositions 694:705 arg1 level 667:671 the level 663:671 the level of individual glycan compositions 663:705 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 0 28 theme Immunoglobulin 14:27 arg1 Biopharmaceuticals 37:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 5 29 theme mannose 761:767 arg1 digestion 743:751 a strong preferential digestion 721:751 a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms 721:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 29 theme mannose 761:767 arg1 result 713:718 a result 711:718 a result 711:718 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 2 30 theme serine 310:315 arg1 essential 344:352 essential 344:352 essential 344:352 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 2 30 theme serine 310:315 arg1 trypsin 326:332 the serine protease trypsin 306:332 the serine protease trypsin 306:332 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 1 31 theme proteolytic 210:220 arg1 cleavage 222:229 the proteolytic cleavage 206:229 the proteolytic cleavage of proteins 206:241 It has been reported that glycosylation can influence the proteolytic cleavage of proteins. 26244886 0 32 theme Biopharmaceuticals 37:54 arg1 Glycoforms 0:9 Glycoforms 0:9 Glycoforms of Immunoglobulin G Based Biopharmaceuticals 0:54 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 6 33 theme antibodies 994:1003 arg1 structure 977:985 the intact higher order structure 953:985 the intact higher order structure of the antibodies 953:1003 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 2 34 theme thorough 246:253 arg1 investigation 255:267 A thorough investigation 244:267 A thorough investigation of this phenomenon 244:286 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 7 35 theme reduced 1172:1178 arg1 time 1189:1192 reduced hands-on time 1172:1192 reduced hands-on time 1172:1192 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 0 36 theme Based 31:35 arg1 Biopharmaceuticals 37:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 0 37 theme G 29:29 arg1 Biopharmaceuticals 37:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Immunoglobulin G Based Biopharmaceuticals 14:54 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 5 38 theme bisected 802:809 arg1 mannose 761:767 high mannose 756:767 high mannose 756:767 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 38 theme bisected 802:809 arg1 glycoforms 811:820 alpha2-3-sialylated and bisected glycoforms 778:820 alpha2-3-sialylated and bisected glycoforms 778:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 3 39 theme G 425:425 arg1 substances 469:478 model substances 463:478 model substances 463:478 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 3 39 theme G 425:425 arg1 biopharmaceuticals 427:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals 384:444 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 4 40 theme glycopeptides 568:580 arg1 quantitation 552:563 Relative quantitation 543:563 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site 543:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 5 41 theme strong 723:728 arg1 digestion 743:751 a strong preferential digestion 721:751 a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms 721:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 41 theme strong 723:728 arg1 result 713:718 a result 711:718 a result 711:718 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 4 42 gly Fc-glycosylation 609:624 arg2 site 626:629 the conserved Fc-glycosylation site 595:629 the conserved Fc-glycosylation site 595:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 3 43 theme bioanalytical 515:527 arg1 applications 529:540 the bioanalytical applications 511:540 the bioanalytical applications 511:540 Monoclonal and polyclonal immunoglobulin G biopharmaceuticals were employed as model substances, which are highly relevant for the bioanalytical applications. 26244886 5 44 theme neutral 858:864 arg1 glycoforms 879:888 the most abundant neutral, fucosylated glycoforms 840:888 the most abundant neutral, fucosylated glycoforms 840:888 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 45 theme preferential 730:741 arg1 digestion 743:751 a strong preferential digestion 721:751 a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms 721:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 45 theme preferential 730:741 arg1 result 713:718 a result 711:718 a result 711:718 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 0 46 from Influence 115:123 arg1 Structure 141:149 Higher-Order Structure 128:149 Higher-Order Structure 128:149 Glycoforms of Immunoglobulin G Based Biopharmaceuticals Are Differentially Cleaved by Trypsin Due to the Glycoform Influence on Higher-Order Structure. 26244886 6 47 theme order 971:975 arg1 structure 977:985 the intact higher order structure 953:985 the intact higher order structure of the antibodies 953:1003 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 7 48 theme hands-on 1180:1187 arg1 time 1189:1192 reduced hands-on time 1172:1192 reduced hands-on time 1172:1192 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 6 49 dep dependent 940:948 arg1 reduced 1040:1046 reduced 1040:1046 was drastically reduced in denatured versus intact antibodies 1024:1084 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26244886 2 50 theme phenomenon 277:286 arg1 investigation 255:267 A thorough investigation 244:267 A thorough investigation of this phenomenon 244:286 A thorough investigation of this phenomenon was conducted for the serine protease trypsin, which is essential in many proteomics workflows. 26244886 1 51 theme proteins 234:241 arg1 cleavage 222:229 the proteolytic cleavage 206:229 the proteolytic cleavage of proteins 206:241 It has been reported that glycosylation can influence the proteolytic cleavage of proteins. 26244886 4 52 gly glycopeptides 568:580 arg2 glycopeptides 568:580 glycopeptides 568:580 glycopeptides derived from the conserved Fc-glycosylation site 568:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 7 53 with protocol 1111:1118 arg1 denaturation 1132:1143 acidic denaturation 1125:1143 acidic denaturation 1125:1143 In addition, a cleavage protocol with acidic denaturation was tested, which featured reduced hands-on time and toxicity while showing highly comparable results to a published denaturation, reduction, and alkylation based protocol. 26244886 4 54 theme individual 676:685 arg1 compositions 694:705 individual glycan compositions 676:705 individual glycan compositions 676:705 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 5 55 theme abundant 849:856 arg1 glycoforms 879:888 the most abundant neutral, fucosylated glycoforms 840:888 the most abundant neutral, fucosylated glycoforms 840:888 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 4 56 theme Relative 543:550 arg1 quantitation 552:563 Relative quantitation 543:563 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site 543:629 Relative quantitation of glycopeptides derived from the conserved Fc-glycosylation site allowed resolution of biases on the level of individual glycan compositions. 26244886 5 57 theme high 756:759 arg1 mannose 761:767 high mannose 756:767 high mannose 756:767 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 57 theme high 756:759 arg1 glycoforms 811:820 alpha2-3-sialylated and bisected glycoforms 778:820 alpha2-3-sialylated and bisected glycoforms 778:820 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 5 57 theme high 756:759 arg1 hybrid 770:775 hybrid 770:775 hybrid 770:775 As a result, a strong preferential digestion of high mannose, hybrid, alpha2-3-sialylated and bisected glycoforms was observed over the most abundant neutral, fucosylated glycoforms. 26244886 6 58 theme intact 1068:1073 arg1 antibodies 1075:1084 denatured versus intact antibodies 1051:1084 denatured versus intact antibodies 1051:1084 Interestingly, this bias was, to a large extent, dependent on the intact higher order structure of the antibodies and, consequently, was drastically reduced in denatured versus intact antibodies. 26220953 0 0 theme Seed 111:114 arg1 Mucilage 116:123 Arabidopsis Seed Mucilage 99:123 Arabidopsis Seed Mucilage 99:123 MUCILAGE-RELATED10 Produces Galactoglucomannan That Maintains Pectin and Cellulose Architecture in Arabidopsis Seed Mucilage. 26220953 11 1 theme GGM 1663:1665 arg1 synthesis 1667:1675 GGM synthesis 1663:1675 GGM synthesis 1663:1675 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 8 2 with glucomannan 1096:1106 arg1 residues 1167:1174 galactose residues 1157:1174 galactose residues 1157:1174 MUCI10 likely decorates glucomannan, synthesized by CELLULOSE SYNTHASE-LIKE A2, with galactose residues in vivo. 26220953 6 3 theme in 854:855 arg1 activity 905:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 12 4 from improvements 1808:1819 arg1 production 1828:1837 the production 1824:1837 the production of valuable commodities from softwoods 1824:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 10 5 theme SCE 1462:1464 arg1 cells 1466:1470 Arabidopsis SCE cells 1450:1470 Arabidopsis SCE cells 1450:1470 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 0 6 theme Arabidopsis 99:109 arg1 Mucilage 116:123 Arabidopsis Seed Mucilage 99:123 Arabidopsis Seed Mucilage 99:123 MUCILAGE-RELATED10 Produces Galactoglucomannan That Maintains Pectin and Cellulose Architecture in Arabidopsis Seed Mucilage. 26220953 10 7 theme Arabidopsis 1450:1460 arg1 cells 1466:1470 Arabidopsis SCE cells 1450:1470 Arabidopsis SCE cells 1450:1470 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 9 8 theme galactosylation 1199:1213 arg1 essential 1218:1226 essential 1218:1226 essential 1218:1226 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 8 theme galactosylation 1199:1213 arg1 degree 1189:1194 The degree 1185:1194 The degree of galactosylation 1185:1213 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 8 theme galactosylation 1199:1213 arg1 density 1304:1310 mucilage density 1295:1310 mucilage density 1295:1310 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 8 theme galactosylation 1199:1213 arg1 structure 1271:1279 the structure 1267:1279 the structure of cellulose 1267:1292 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 8 theme galactosylation 1199:1213 arg1 adherence 1328:1336 the adherence 1324:1336 the adherence of pectin 1324:1346 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 3 9 theme cell 514:517 arg1 synthesis 524:532 cell wall synthesis 514:532 cell wall synthesis 514:532 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 10 10 dep synthesis 1523:1531 arg1 the 1519:1521 the 1519:1521 the 1519:1521 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 4 11 theme secondary 577:585 arg1 rich 600:603 rich 600:603 rich 600:603 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 11 theme secondary 577:585 arg1 wall 587:590 a specialized secondary wall 563:590 a specialized secondary wall that is rich in pectin, at a precise stage of development 563:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 11 theme secondary 577:585 arg1 mucilage 553:560 mucilage 553:560 mucilage 553:560 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 10 12 theme mucilage 1387:1394 arg1 architecture 1396:1407 mucilage architecture 1387:1407 mucilage architecture 1387:1407 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 12 13 from production 1828:1837 arg1 softwoods 1868:1876 softwoods 1868:1876 softwoods 1868:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 6 14 theme α-1,6-galactosyltransferase 877:903 arg1 activity 905:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 7 15 theme mutants 951:957 arg1 analysis 928:935 Our detailed analysis 915:935 Our detailed analysis of the muci10 mutants 915:957 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 6 16 contain has 850:852 arg1 enzyme 838:843 a fenugreek (Trigonella foenumgraecum) enzyme 799:843 a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity 799:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 6 16 contain has 850:852 arg2 activity 905:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 5 17 theme mucilage 749:756 arg1 properties 758:767 mucilage properties 749:767 mucilage properties 749:767 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 6 18 theme galactomannan 863:875 arg1 activity 905:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 in vitro galactomannan α-1,6-galactosyltransferase activity 854:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 10 19 theme excellent 1485:1493 arg1 model 1495:1499 an excellent model 1482:1499 an excellent model in which to study the synthesis and function of GGM 1482:1551 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 8 20 theme galactose 1157:1165 arg1 residues 1167:1174 galactose residues 1157:1174 galactose residues 1157:1174 MUCI10 likely decorates glucomannan, synthesized by CELLULOSE SYNTHASE-LIKE A2, with galactose residues in vivo. 26220953 10 21 theme GGM 1549:1551 arg1 function 1537:1544 function 1537:1544 function 1537:1544 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 10 21 theme GGM 1549:1551 arg1 synthesis 1523:1531 synthesis 1523:1531 synthesis 1523:1531 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 1 22 theme extracellular 191:203 arg1 built 212:216 an extracellular matrix built 188:216 an extracellular matrix built of polysaccharides 188:235 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 3 23 theme Arabidopsis 457:467 arg1 thaliana 469:476 Arabidopsis thaliana 457:476 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) 440:503 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 12 24 theme abundant 1700:1707 arg1 hemicellulose 1709:1721 the most abundant hemicellulose 1691:1721 the most abundant hemicellulose in the secondary walls of gymnosperms 1691:1759 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 12 24 theme abundant 1700:1707 arg1 GGM 1684:1686 GGM 1684:1686 GGM 1684:1686 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 11 25 with varieties 1574:1582 arg1 defects 1589:1595 defects 1589:1595 defects similar to muci10 mutants 1589:1621 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 3 26 theme thaliana 469:476 arg1 SCE 500:502 SCE 500:502 SCE 500:502 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 3 26 theme thaliana 469:476 arg1 epidermis 489:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis 440:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) 440:503 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 4 27 from rich 600:603 arg1 pectin 608:613 pectin 608:613 pectin 608:613 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 3 28 theme Arabidopsis 444:454 arg1 SCE 500:502 SCE 500:502 SCE 500:502 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 3 28 theme Arabidopsis 444:454 arg1 epidermis 489:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis 440:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) 440:503 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 6 29 theme fenugreek 801:809 arg1 enzyme 838:843 a fenugreek (Trigonella foenumgraecum) enzyme 799:843 a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity 799:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 10 30 theme cellulosic 1420:1429 arg1 rays 1431:1434 cellulosic rays 1420:1434 cellulosic rays 1420:1434 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 4 31 from stage 629:633 arg1 mucilage 553:560 mucilage 553:560 mucilage 553:560 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 31 from stage 629:633 arg1 wall 587:590 a specialized secondary wall 563:590 a specialized secondary wall that is rich in pectin, at a precise stage of development 563:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 31 from stage 629:633 arg1 rich 600:603 rich 600:603 rich 600:603 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 1 32 theme matrix 205:210 arg1 built 212:216 an extracellular matrix built 188:216 an extracellular matrix built of polysaccharides 188:235 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 9 33 theme pectin 1341:1346 arg1 essential 1218:1226 essential 1218:1226 essential 1218:1226 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 33 theme pectin 1341:1346 arg1 degree 1189:1194 The degree 1185:1194 The degree of galactosylation 1185:1213 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 33 theme pectin 1341:1346 arg1 density 1304:1310 mucilage density 1295:1310 mucilage density 1295:1310 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 33 theme pectin 1341:1346 arg1 structure 1271:1279 the structure 1267:1279 the structure of cellulose 1267:1292 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 33 theme pectin 1341:1346 arg1 adherence 1328:1336 the adherence 1324:1336 the adherence of pectin 1324:1346 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 11 34 theme natural 1566:1572 arg1 varieties 1574:1582 Arabidopsis natural varieties 1554:1582 Arabidopsis natural varieties with defects similar to muci10 mutants 1554:1621 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 3 35 theme coat 484:487 arg1 SCE 500:502 SCE 500:502 SCE 500:502 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 3 35 theme coat 484:487 arg1 epidermis 489:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis 440:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) 440:503 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 10 36 theme GGM 1365:1367 arg1 scaffolds 1369:1377 GGM scaffolds 1365:1377 GGM scaffolds 1365:1377 We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. 26220953 4 37 theme specialized 565:575 arg1 rich 600:603 rich 600:603 rich 600:603 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 37 theme specialized 565:575 arg1 wall 587:590 a specialized secondary wall 563:590 a specialized secondary wall that is rich in pectin, at a precise stage of development 563:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 37 theme specialized 565:575 arg1 mucilage 553:560 mucilage 553:560 mucilage 553:560 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 5 38 theme MUCI 695:698 arg1 genes 701:705 MUCILAGE-RELATED (MUCI) genes 677:705 MUCILAGE-RELATED (MUCI) genes 677:705 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 9 39 theme GGM 1253:1255 arg1 backbone 1257:1264 the GGM backbone 1249:1264 the GGM backbone 1249:1264 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 7 40 theme unbranched 1048:1057 arg1 glucomannan 1059:1069 unbranched glucomannan 1048:1069 highly branched galactoglucomannan (GGM) rather than unbranched glucomannan 995:1069 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 1 41 theme resources 155:163 arg1 lot 142:144 a lot 140:144 a lot of their resources into the production of an extracellular matrix built of polysaccharides 140:235 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 1 41 theme resources 155:163 arg1 resources 155:163 their resources 149:163 their resources 149:163 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 5 42 theme properties 758:767 arg1 determinant 734:744 a key determinant 728:744 a key determinant of mucilage properties 728:767 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 5 42 theme properties 758:767 arg1 MUCI10 718:723 MUCI10 718:723 MUCI10 718:723 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 8 43 theme CELLULOSE 1124:1132 arg1 A2 1148:1149 CELLULOSE SYNTHASE-LIKE A2 1124:1149 CELLULOSE SYNTHASE-LIKE A2 1124:1149 MUCI10 likely decorates glucomannan, synthesized by CELLULOSE SYNTHASE-LIKE A2, with galactose residues in vivo. 26220953 9 44 theme backbone 1257:1264 arg1 synthesis 1236:1244 the synthesis 1232:1244 the synthesis of the GGM backbone 1232:1264 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 1 45 theme polysaccharides 221:235 arg1 built 212:216 an extracellular matrix built 188:216 an extracellular matrix built of polysaccharides 188:235 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 4 46 theme development 638:648 arg1 stage 629:633 a precise stage 619:633 a precise stage of development 619:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 47 theme SCE 535:537 arg1 cells 539:543 SCE cells 535:543 SCE cells 535:543 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 11 48 theme Arabidopsis 1554:1564 arg1 varieties 1574:1582 Arabidopsis natural varieties 1554:1582 Arabidopsis natural varieties with defects similar to muci10 mutants 1554:1621 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 12 49 theme valuable 1842:1849 arg1 commodities 1851:1861 valuable commodities 1842:1861 valuable commodities from softwoods 1842:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 3 50 theme seed 479:482 arg1 SCE 500:502 SCE 500:502 SCE 500:502 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 3 50 theme seed 479:482 arg1 epidermis 489:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis 440:497 the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) 440:503 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 0 51 theme Cellulose 73:81 arg1 Architecture 83:94 Cellulose Architecture 73:94 Cellulose Architecture 73:94 MUCILAGE-RELATED10 Produces Galactoglucomannan That Maintains Pectin and Cellulose Architecture in Arabidopsis Seed Mucilage. 26220953 5 52 theme coexpression 653:664 arg1 search 666:671 A coexpression search 651:671 A coexpression search for MUCILAGE-RELATED (MUCI) genes 651:705 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 7 53 contain contains 986:993 arg2 GGM 1031:1033 GGM 1031:1033 GGM 1031:1033 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 7 53 contain contains 986:993 arg2 glucomannan 1059:1069 unbranched glucomannan 1048:1069 highly branched galactoglucomannan (GGM) rather than unbranched glucomannan 995:1069 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 7 53 contain contains 986:993 arg2 galactoglucomannan 1011:1028 highly branched galactoglucomannan 995:1028 highly branched galactoglucomannan (GGM) rather than unbranched glucomannan 995:1069 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 7 53 contain contains 986:993 arg1 mucilage 977:984 mucilage 977:984 mucilage 977:984 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 11 54 theme muci10 1608:1613 arg1 mutants 1615:1621 muci10 mutants 1608:1621 muci10 mutants 1608:1621 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 12 55 theme gymnosperms 1749:1759 arg1 walls 1740:1744 the secondary walls 1726:1744 the secondary walls of gymnosperms 1726:1759 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 12 56 from softwoods 1868:1876 arg1 production 1828:1837 the production 1824:1837 the production of valuable commodities from softwoods 1824:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 12 56 from softwoods 1868:1876 arg1 commodities 1851:1861 valuable commodities 1842:1861 valuable commodities from softwoods 1842:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 8 57 theme SYNTHASE-LIKE 1134:1146 arg1 A2 1148:1149 CELLULOSE SYNTHASE-LIKE A2 1124:1149 CELLULOSE SYNTHASE-LIKE A2 1124:1149 MUCI10 likely decorates glucomannan, synthesized by CELLULOSE SYNTHASE-LIKE A2, with galactose residues in vivo. 26220953 6 58 theme foenumgraecum 823:835 arg1 enzyme 838:843 a fenugreek (Trigonella foenumgraecum) enzyme 799:843 a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity 799:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 12 59 from hemicellulose 1709:1721 arg1 walls 1740:1744 the secondary walls 1726:1744 the secondary walls of gymnosperms 1726:1759 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 12 60 theme commodities 1851:1861 arg1 production 1828:1837 the production 1824:1837 the production of valuable commodities from softwoods 1824:1876 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 9 61 dep essential 1218:1226 arg1 essential 1218:1226 essential 1218:1226 essential 1218:1226 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 61 dep essential 1218:1226 arg1 degree 1189:1194 The degree 1185:1194 The degree of galactosylation 1185:1213 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 61 dep essential 1218:1226 arg1 density 1304:1310 mucilage density 1295:1310 mucilage density 1295:1310 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 61 dep essential 1218:1226 arg1 structure 1271:1279 the structure 1267:1279 the structure of cellulose 1267:1292 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 61 dep essential 1218:1226 arg1 adherence 1328:1336 the adherence 1324:1336 the adherence of pectin 1324:1346 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 11 62 theme additional 1634:1643 arg1 genes 1645:1649 additional genes 1634:1649 additional genes involved in GGM synthesis 1634:1675 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 6 63 theme Trigonella 812:821 arg1 enzyme 838:843 a fenugreek (Trigonella foenumgraecum) enzyme 799:843 a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity 799:912 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 11 64 theme similar 1597:1603 arg1 defects 1589:1595 defects 1589:1595 defects similar to muci10 mutants 1589:1621 Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. 26220953 2 65 theme polymers 343:350 arg1 functions 315:323 the functions 311:323 the functions of the individual polymers 311:350 While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. 26220953 2 65 theme polymers 343:350 arg1 enzymes 360:366 the enzymes 356:366 the enzymes that catalyze their biosynthesis 356:399 While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. 26220953 7 66 theme detailed 919:926 arg1 analysis 928:935 Our detailed analysis 915:935 Our detailed analysis of the muci10 mutants 915:957 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 5 67 theme MUCILAGE-RELATED 677:692 arg1 genes 701:705 MUCILAGE-RELATED (MUCI) genes 677:705 MUCILAGE-RELATED (MUCI) genes 677:705 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 9 68 theme cellulose 1284:1292 arg1 essential 1218:1226 essential 1218:1226 essential 1218:1226 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 68 theme cellulose 1284:1292 arg1 degree 1189:1194 The degree 1185:1194 The degree of galactosylation 1185:1213 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 68 theme cellulose 1284:1292 arg1 density 1304:1310 mucilage density 1295:1310 mucilage density 1295:1310 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 68 theme cellulose 1284:1292 arg1 structure 1271:1279 the structure 1267:1279 the structure of cellulose 1267:1292 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 68 theme cellulose 1284:1292 arg1 adherence 1328:1336 the adherence 1324:1336 the adherence of pectin 1324:1346 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 2 69 theme wall 272:275 arg1 composition 248:258 the composition 244:258 the composition of the cell wall 244:275 While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. 26220953 7 70 theme branched 1002:1009 arg1 GGM 1031:1033 GGM 1031:1033 GGM 1031:1033 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 7 70 theme branched 1002:1009 arg1 galactoglucomannan 1011:1028 highly branched galactoglucomannan 995:1028 highly branched galactoglucomannan (GGM) rather than unbranched glucomannan 995:1069 Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. 26220953 2 71 theme individual 332:341 arg1 polymers 343:350 the individual polymers 328:350 the individual polymers 328:350 While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. 26220953 12 72 theme secondary 1730:1738 arg1 walls 1740:1744 the secondary walls 1726:1744 the secondary walls of gymnosperms 1726:1759 Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. 26220953 6 73 dep in 854:855 arg1 vitro 857:861 vitro 857:861 vitro 857:861 MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. 26220953 5 74 theme key 730:732 arg1 determinant 734:744 a key determinant 728:744 a key determinant of mucilage properties 728:767 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 5 74 theme key 730:732 arg1 MUCI10 718:723 MUCI10 718:723 MUCI10 718:723 A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. 26220953 4 75 theme precise 621:627 arg1 stage 629:633 a precise stage 619:633 a precise stage of development 619:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 2 76 theme cell 267:270 arg1 wall 272:275 the cell wall 263:275 the cell wall 263:275 While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. 26220953 1 77 theme built 212:216 arg1 production 174:183 the production 170:183 the production of an extracellular matrix built of polysaccharides 170:235 Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. 26220953 3 78 theme wall 519:522 arg1 synthesis 524:532 cell wall synthesis 514:532 cell wall synthesis 514:532 We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. 26220953 4 79 from pectin 608:613 arg1 mucilage 553:560 mucilage 553:560 mucilage 553:560 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 79 from pectin 608:613 arg1 wall 587:590 a specialized secondary wall 563:590 a specialized secondary wall that is rich in pectin, at a precise stage of development 563:648 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 4 79 from pectin 608:613 arg1 rich 600:603 rich 600:603 rich 600:603 SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. 26220953 9 80 theme mucilage 1295:1302 arg1 essential 1218:1226 essential 1218:1226 essential 1218:1226 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 80 theme mucilage 1295:1302 arg1 degree 1189:1194 The degree 1185:1194 The degree of galactosylation 1185:1213 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 80 theme mucilage 1295:1302 arg1 density 1304:1310 mucilage density 1295:1310 mucilage density 1295:1310 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 80 theme mucilage 1295:1302 arg1 structure 1271:1279 the structure 1267:1279 the structure of cellulose 1267:1292 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 26220953 9 80 theme mucilage 1295:1302 arg1 adherence 1328:1336 the adherence 1324:1336 the adherence of pectin 1324:1346 The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. 25108239 0 0 theme activation 82:91 arg1 mechanism 93:101 activation mechanism 82:101 activation mechanism 82:101 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 6 1 theme sn-3 955:958 arg1 acids 966:970 sn-3 fatty acids 955:970 sn-3 fatty acids 955:970 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 7 2 theme internal 998:1005 arg1 One 980:982 One 980:982 One 980:982 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 7 2 theme internal 998:1005 arg1 tunnel 1007:1012 an internal tunnel 995:1012 an internal tunnel 995:1012 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 7 2 theme internal 998:1005 arg1 them 987:990 them 987:990 them 987:990 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 8 3 theme structural 1130:1139 arg1 results 1157:1163 our structural and biochemical results 1126:1163 our structural and biochemical results 1126:1163 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 5 4 theme structural 660:669 arg1 rearrangements 671:684 structural rearrangements 660:684 structural rearrangements of loop α16-α17 660:700 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 6 5 theme acids 966:970 arg1 chains 972:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 6 6 theme hole 892:895 arg1 position 867:874 the position 863:874 the position of the oxyanion hole 863:895 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 2 7 theme high 292:295 arg1 versatility 297:307 Their high versatility 286:307 Their high versatility 286:307 Their high versatility makes them excellent candidates for biotechnological purposes. 25108239 6 8 theme fatty 960:964 arg1 acids 966:970 sn-3 fatty acids 955:970 sn-3 fatty acids 955:970 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 6 9 theme oxyanion 883:890 arg1 hole 892:895 the oxyanion hole 879:895 the oxyanion hole 879:895 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 7 10 theme them 987:990 arg1 One 980:982 One 980:982 One 980:982 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 7 10 theme them 987:990 arg1 tunnel 1007:1012 an internal tunnel 995:1012 an internal tunnel 995:1012 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 7 10 theme them 987:990 arg1 them 987:990 them 987:990 them 987:990 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 5 11 theme large 619:623 arg1 displacement 625:636 a large displacement 617:636 a large displacement of the conserved lid 617:657 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 1 12 theme organic 268:274 arg1 solvents 276:283 organic solvents 268:283 organic solvents 268:283 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 5 13 theme large 735:739 arg1 opening 741:747 a large opening 733:747 a large opening 733:747 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 14 with dimer 722:726 arg1 opening 741:747 a large opening 733:747 a large opening 733:747 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 0 15 theme product 107:113 arg1 release 115:121 product release 107:121 product release 107:121 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 3 16 from piceae 411:416 arg1 esterase 379:386 Sterol esterase 372:386 Sterol esterase from fungus Ophiostoma piceae (OPE) 372:422 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 7 17 theme outer 1053:1057 arg1 surface 1059:1065 the outer surface 1049:1065 the outer surface of the enzyme 30 Å far from the catalytic Ser220 1049:1114 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 1 18 theme sterol 180:185 arg1 esters 187:192 sterol esters 180:192 sterol esters 180:192 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 4 19 theme Crystal 499:505 arg1 structures 507:516 Crystal structures 499:516 Crystal structures of OPE 499:523 Crystal structures of OPE were solved in this study for the closed and open conformations. 25108239 5 20 theme conserved 645:653 arg1 lid 655:657 the conserved lid 641:657 the conserved lid 641:657 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 4 21 theme closed 559:564 arg1 conformations 575:587 the closed and open conformations 555:587 the closed and open conformations 555:587 Crystal structures of OPE were solved in this study for the closed and open conformations. 25108239 2 22 theme excellent 320:328 arg1 candidates 330:339 excellent candidates 320:339 excellent candidates for biotechnological purposes 320:369 Their high versatility makes them excellent candidates for biotechnological purposes. 25108239 0 23 theme Crystal 0:6 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 0:122 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 1 24 theme solvents 276:283 arg1 presence 256:263 the presence 252:263 the presence of organic solvents 252:283 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 5 25 theme lid 655:657 arg1 rearrangements 671:684 structural rearrangements 660:684 structural rearrangements of loop α16-α17 660:700 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 25 theme lid 655:657 arg1 formation 707:715 formation 707:715 formation of a dimer with a large opening 707:747 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 25 theme lid 655:657 arg1 displacement 625:636 a large displacement 617:636 a large displacement of the conserved lid 617:657 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 26 theme Enzyme 590:595 arg1 activation 597:606 Enzyme activation 590:606 Enzyme activation 590:606 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 6 27 theme triglyceride 825:836 arg1 substrate 838:846 the triglyceride substrate 821:846 the triglyceride substrate 821:846 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 3 28 theme family 439:444 arg1 abH03.01 446:453 the family abH03.01 435:453 the family abH03.01 of the Candida rugosa lipase-like proteins 435:496 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 0 29 theme Ophiostoma 22:31 arg1 esterase 47:54 Ophiostoma piceae sterol esterase 22:54 Ophiostoma piceae sterol esterase 22:54 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 6 30 theme sn-2 946:949 arg1 chains 972:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 6 31 theme active 788:793 arg1 site 795:798 the active site 784:798 the active site 784:798 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 1 32 dep able 145:148 arg1 carry 219:223 carry 219:223 to carry out synthesis reactions in the presence of organic solvents 216:283 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 1 32 dep able 145:148 arg1 hydrolyze 165:173 hydrolyze 165:173 to efficiently hydrolyze both sterol esters and triglycerides 150:210 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 3 33 theme rugosa 470:475 arg1 proteins 489:496 the Candida rugosa lipase-like proteins 458:496 the Candida rugosa lipase-like proteins 458:496 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 3 34 theme Ophiostoma 400:409 arg1 piceae 411:416 fungus Ophiostoma piceae 393:416 fungus Ophiostoma piceae (OPE) 393:422 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 3 34 theme Ophiostoma 400:409 arg1 OPE 419:421 OPE 419:421 OPE 419:421 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 4 35 theme open 570:573 arg1 conformations 575:587 the closed and open conformations 555:587 the closed and open conformations 555:587 Crystal structures of OPE were solved in this study for the closed and open conformations. 25108239 7 36 theme active 1030:1035 arg1 center 1037:1042 the active center 1026:1042 the active center 1026:1042 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 3 37 theme lipase-like 477:487 arg1 proteins 489:496 the Candida rugosa lipase-like proteins 458:496 the Candida rugosa lipase-like proteins 458:496 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 0 38 theme sterol 40:45 arg1 esterase 47:54 Ophiostoma piceae sterol esterase 22:54 Ophiostoma piceae sterol esterase 22:54 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 2 39 theme biotechnological 345:360 arg1 purposes 362:369 biotechnological purposes 345:369 biotechnological purposes 345:369 Their high versatility makes them excellent candidates for biotechnological purposes. 25108239 8 40 theme enzymes 1359:1365 arg1 properties 1339:1348 the catalytic properties 1325:1348 the catalytic properties of these enzymes 1325:1365 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 6 41 theme sn-1 940:943 arg1 chains 972:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 the sn-1, sn-2 and sn-3 fatty acids chains 936:977 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 0 42 theme piceae 33:38 arg1 esterase 47:54 Ophiostoma piceae sterol esterase 22:54 Ophiostoma piceae sterol esterase 22:54 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 5 43 theme dimer 722:726 arg1 rearrangements 671:684 structural rearrangements 660:684 structural rearrangements of loop α16-α17 660:700 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 43 theme dimer 722:726 arg1 formation 707:715 formation 707:715 formation of a dimer with a large opening 707:747 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 43 theme dimer 722:726 arg1 displacement 625:636 a large displacement 617:636 a large displacement of the conserved lid 617:657 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 8 44 theme biotechnological 1377:1392 arg1 applications 1394:1405 their biotechnological applications 1371:1405 their biotechnological applications 1371:1405 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 3 45 theme fungus 393:398 arg1 piceae 411:416 fungus Ophiostoma piceae 393:416 fungus Ophiostoma piceae (OPE) 393:422 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 3 45 theme fungus 393:398 arg1 OPE 419:421 OPE 419:421 OPE 419:421 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 3 46 theme Candida 462:468 arg1 proteins 489:496 the Candida rugosa lipase-like proteins 458:496 the Candida rugosa lipase-like proteins 458:496 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 7 47 theme enzyme 1074:1079 arg1 Å 1084:1084 the enzyme 30 Å 1070:1084 the enzyme 30 Å far from the catalytic Ser220 1070:1114 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 5 48 theme loop 689:692 arg1 rearrangements 671:684 structural rearrangements 660:684 structural rearrangements of loop α16-α17 660:700 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 48 theme loop 689:692 arg1 formation 707:715 formation 707:715 formation of a dimer with a large opening 707:747 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 5 48 theme loop 689:692 arg1 displacement 625:636 a large displacement 617:636 a large displacement of the conserved lid 617:657 Enzyme activation involves a large displacement of the conserved lid, structural rearrangements of loop α16-α17, and formation of a dimer with a large opening. 25108239 6 49 theme substrate 838:846 arg1 chains 811:816 chains 811:816 chains 811:816 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 6 49 theme substrate 838:846 arg1 pockets 911:917 the three pockets 901:917 the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains 901:977 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 6 50 theme PEG 756:758 arg1 molecules 760:768 Three PEG molecules 750:768 Three PEG molecules 750:768 Three PEG molecules are placed in the active site, mimicking chains of the triglyceride substrate, demonstrating the position of the oxyanion hole and the three pockets that accommodate the sn-1, sn-2 and sn-3 fatty acids chains. 25108239 8 51 theme different 1216:1224 arg1 substrates 1226:1235 different substrates 1216:1235 different substrates 1216:1235 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 0 52 theme esterase 47:54 arg1 structures 8:17 Crystal structures 0:17 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 0:122 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 3 53 theme Sterol 372:377 arg1 esterase 379:386 Sterol esterase 372:386 Sterol esterase from fungus Ophiostoma piceae (OPE) 372:422 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 1 54 contain carry 219:223 arg1 esterases 131:139 Sterol esterases 124:139 Sterol esterases 124:139 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 1 54 contain carry 219:223 arg3 presence 256:263 the presence 252:263 the presence of organic solvents 252:283 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 1 54 contain carry 219:223 arg2 reactions 239:247 synthesis reactions 229:247 synthesis reactions 229:247 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 1 55 theme Sterol 124:129 arg1 esterases 131:139 Sterol esterases 124:139 Sterol esterases 124:139 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 7 56 theme Å 1084:1084 arg1 surface 1059:1065 the outer surface 1049:1065 the outer surface of the enzyme 30 Å far from the catalytic Ser220 1049:1114 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 25108239 8 57 theme variants 1305:1312 arg1 construction 1285:1296 the construction 1281:1296 the construction of new variants 1281:1312 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 3 58 theme proteins 489:496 arg1 abH03.01 446:453 the family abH03.01 435:453 the family abH03.01 of the Candida rugosa lipase-like proteins 435:496 Sterol esterase from fungus Ophiostoma piceae (OPE) belongs to the family abH03.01 of the Candida rugosa lipase-like proteins. 25108239 8 59 theme new 1301:1303 arg1 variants 1305:1312 new variants 1301:1312 new variants 1301:1312 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 8 60 theme biochemical 1145:1155 arg1 results 1157:1163 our structural and biochemical results 1126:1163 our structural and biochemical results 1126:1163 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 8 61 theme substrates 1226:1235 arg1 substrates 1226:1235 different substrates 1216:1235 different substrates 1216:1235 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 8 61 theme substrates 1226:1235 arg1 variety 1205:1211 a great variety 1197:1211 a great variety of different substrates 1197:1235 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 1 62 theme synthesis 229:237 arg1 reactions 239:247 synthesis reactions 229:247 synthesis reactions 229:247 Sterol esterases are able to efficiently hydrolyze both sterol esters and triglycerides and to carry out synthesis reactions in the presence of organic solvents. 25108239 8 63 theme catalytic 1329:1337 arg1 properties 1339:1348 the catalytic properties 1325:1348 the catalytic properties of these enzymes 1325:1365 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 8 64 theme great 1199:1203 arg1 substrates 1226:1235 different substrates 1216:1235 different substrates 1216:1235 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 8 64 theme great 1199:1203 arg1 variety 1205:1211 a great variety 1197:1211 a great variety of different substrates 1197:1235 Based on our structural and biochemical results we propose a mechanism by which a great variety of different substrates can be hydrolyzed in OPE paving the way for the construction of new variants to improve the catalytic properties of these enzymes and their biotechnological applications. 25108239 0 65 dep structures 8:17 arg1 insights 68:75 structural insights 57:75 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 0:122 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 0 66 theme structural 57:66 arg1 insights 68:75 structural insights 57:75 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 0:122 Crystal structures of Ophiostoma piceae sterol esterase: structural insights into activation mechanism and product release. 25108239 4 67 theme OPE 521:523 arg1 structures 507:516 Crystal structures 499:516 Crystal structures of OPE 499:523 Crystal structures of OPE were solved in this study for the closed and open conformations. 25108239 7 68 theme catalytic 1099:1107 arg1 Ser220 1109:1114 the catalytic Ser220 1095:1114 the catalytic Ser220 1095:1114 One of them is an internal tunnel, connecting the active center with the outer surface of the enzyme 30 Å far from the catalytic Ser220. 28485610 4 0 theme bond 586:589 arg1 cleavage 591:598 An unexpected glycosidic bond cleavage 561:598 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate 561:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 4 1 theme intermediate 664:675 arg1 oxidation 636:644 the TEMPO/PhI(OAc)2-mediated oxidation 607:644 the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate 607:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 4 2 theme disaccharide 651:662 arg1 intermediate 664:675 a disaccharide intermediate 649:675 a disaccharide intermediate 649:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 1 3 dep Staphylococcus 109:122 arg1 aureus 124:129 aureus 124:129 aureus 124:129 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28485610 3 4 theme α-fucosamine 468:479 arg1 linkage 481:487 The α-fucosamine linkage 464:487 The α-fucosamine linkage 464:487 The α-fucosamine linkage was installed stereoselectively, using a reactive 2-azidofucosyl donor. 28485610 4 5 theme glycosidic 575:584 arg1 cleavage 591:598 An unexpected glycosidic bond cleavage 561:598 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate 561:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 2 6 theme linkages 298:305 arg1 construction 252:263 the construction 248:263 the construction of the α-galactosaminuronic acid linkages 248:305 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 4 7 theme unexpected 564:573 arg1 cleavage 591:598 An unexpected glycosidic bond cleavage 561:598 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate 561:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 2 8 theme galactopyranoside 375:391 arg1 donor 393:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 2 9 theme model 442:446 arg1 glycosylations 448:461 model glycosylations 442:461 model glycosylations 442:461 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 2 10 theme 2-azido-4,6-O-di-tert-butylsilylidene 337:373 arg1 donor 393:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 2 11 used utilized 235:242 arg2 strategy 222:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 0 12 theme Staphylococcus 17:30 arg1 Polysaccharide 57:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 4 13 theme 2-mediated 625:634 arg1 oxidation 636:644 the TEMPO/PhI(OAc)2-mediated oxidation 607:644 the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate 607:675 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 2 14 theme stereoselective 321:335 arg1 donor 393:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor 319:397 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 3 15 theme 2-azidofucosyl 539:552 arg1 donor 554:558 a reactive 2-azidofucosyl donor 528:558 a reactive 2-azidofucosyl donor 528:558 The α-fucosamine linkage was installed stereoselectively, using a reactive 2-azidofucosyl donor. 28485610 0 16 theme Strain 39:44 arg1 Polysaccharide 57:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 4 17 theme 2-Pinnick 713:721 arg1 protocol 733:740 a TEMPO/PhI(OAc)2-Pinnick oxidation protocol 697:740 a TEMPO/PhI(OAc)2-Pinnick oxidation protocol 697:740 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 4 18 theme oxidation 723:731 arg1 protocol 733:740 a TEMPO/PhI(OAc)2-Pinnick oxidation protocol 697:740 a TEMPO/PhI(OAc)2-Pinnick oxidation protocol 697:740 An unexpected glycosidic bond cleavage during the TEMPO/PhI(OAc)2-mediated oxidation of a disaccharide intermediate was circumvented by a TEMPO/PhI(OAc)2-Pinnick oxidation protocol. 28485610 3 19 theme reactive 530:537 arg1 donor 554:558 a reactive 2-azidofucosyl donor 528:558 a reactive 2-azidofucosyl donor 528:558 The α-fucosamine linkage was installed stereoselectively, using a reactive 2-azidofucosyl donor. 28485610 0 20 theme Capsular 48:55 arg1 Polysaccharide 57:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 1 21 theme Staphylococcus 109:122 arg1 polysaccharide 149:162 the Staphylococcus aureus strain M capsular polysaccharide 105:162 the Staphylococcus aureus strain M capsular polysaccharide repeating unit 105:177 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28485610 0 22 theme M 46:46 arg1 Polysaccharide 57:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 the Staphylococcus aureus Strain M Capsular Polysaccharide 13:70 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 0 23 dep Staphylococcus 17:30 arg1 aureus 32:37 aureus 32:37 aureus 32:37 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 2 24 theme acid 293:296 arg1 linkages 298:305 the α-galactosaminuronic acid linkages 268:305 the α-galactosaminuronic acid linkages 268:305 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 1 25 theme strain 131:136 arg1 polysaccharide 149:162 the Staphylococcus aureus strain M capsular polysaccharide 105:162 the Staphylococcus aureus strain M capsular polysaccharide repeating unit 105:177 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28485610 2 26 theme oxidation 212:220 arg1 strategy 222:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 2 27 theme α-galactosaminuronic 272:291 arg1 linkages 298:305 the α-galactosaminuronic acid linkages 268:305 the α-galactosaminuronic acid linkages 268:305 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 1 28 theme M 138:138 arg1 polysaccharide 149:162 the Staphylococcus aureus strain M capsular polysaccharide 105:162 the Staphylococcus aureus strain M capsular polysaccharide repeating unit 105:177 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28485610 2 29 theme postglycosylation 194:210 arg1 strategy 222:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy 192:229 A postglycosylation oxidation strategy was utilized for the construction of the α-galactosaminuronic acid linkages, relying on a stereoselective 2-azido-4,6-O-di-tert-butylsilylidene galactopyranoside donor, for which the selectivity was assessed by model glycosylations. 28485610 1 30 theme capsular 140:147 arg1 polysaccharide 149:162 the Staphylococcus aureus strain M capsular polysaccharide 105:162 the Staphylococcus aureus strain M capsular polysaccharide repeating unit 105:177 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28485610 0 31 theme Polysaccharide 57:70 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide 0:70 Synthesis of the Staphylococcus aureus Strain M Capsular Polysaccharide Repeating Unit. 28485610 1 32 theme polysaccharide 149:162 arg1 synthesis 92:100 The synthesis 88:100 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit 88:177 The synthesis of the Staphylococcus aureus strain M capsular polysaccharide repeating unit is reported. 28888681 7 0 theme higher 968:973 arg1 selectivity 975:985 higher selectivity 968:985 higher selectivity 968:985 The nanoHILIC method provided higher selectivity based on the composition of glycan, compared to nanoRP-LC but a lower sensitivity. 28888681 1 1 theme glycosylation 148:160 arg1 Analysis 132:139 Analysis 132:139 Analysis of the glycosylation of proteins 132:172 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 1 1 theme glycosylation 148:160 arg1 challenge 179:187 a challenge 177:187 a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms 177:268 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 6 2 gly glycopeptides 893:905 arg2 glycopeptides 893:905 glycopeptides 893:905 glycopeptides 893:905 The presence of sialic acids prolonged the retention of glycopeptides in both chromatographic modes. 28888681 2 3 theme tools 390:394 arg1 tools 390:394 the most powerful tools 372:394 the most powerful tools for glycopeptide analysis 372:420 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 3 theme tools 390:394 arg1 one 365:367 one 365:367 one 365:367 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 3 theme tools 390:394 arg1 combination 273:283 A combination 271:283 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) 271:360 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 7 4 theme nanoHILIC 942:950 arg1 method 952:957 The nanoHILIC method 938:957 The nanoHILIC method 938:957 The nanoHILIC method provided higher selectivity based on the composition of glycan, compared to nanoRP-LC but a lower sensitivity. 28888681 8 5 theme fucosylated 1289:1299 arg1 glycoforms 1301:1310 the fucosylated glycoforms 1285:1310 the fucosylated glycoforms 1285:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 1 6 theme diverse 251:257 arg1 glycoforms 259:268 the diverse glycoforms 247:268 the diverse glycoforms 247:268 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 8 7 theme fucose 1141:1146 arg1 isomers 1130:1136 separate linkage isomers 1113:1136 separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1113:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 8 dep fucose 1141:1146 arg1 arm 1164:1166 outer arm 1158:1166 outer arm 1158:1166 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 8 dep fucose 1141:1146 arg1 core 1149:1152 core 1149:1152 core 1149:1152 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 2 9 theme reversed 288:295 arg1 chromatography 303:316 reversed phase chromatography 288:316 reversed phase chromatography 288:316 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 1 10 theme glycoforms 259:268 arg1 separation 233:242 separation 233:242 separation of the diverse glycoforms 233:268 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 2 11 gly glycopeptide 400:411 arg2 glycopeptide 400:411 glycopeptide analysis 400:420 glycopeptide analysis 400:420 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 3 12 theme liquid 497:502 arg1 chromatography 504:517 hydrophilic interaction liquid chromatography 473:517 hydrophilic interaction liquid chromatography (HILIC) 473:525 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 3 12 theme liquid 497:502 arg1 HILIC 520:524 HILIC 520:524 HILIC 520:524 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 4 13 theme retention 694:702 arg1 time 704:707 the retention time 690:707 the retention time 690:707 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 6 14 theme acids 860:864 arg1 presence 841:848 The presence 837:848 The presence of sialic acids 837:864 The presence of sialic acids prolonged the retention of glycopeptides in both chromatographic modes. 28888681 8 15 from elucidation 1253:1263 arg1 beneficial 1235:1244 beneficial 1235:1244 beneficial 1235:1244 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 1 16 theme proteins 165:172 arg1 glycosylation 148:160 the glycosylation 144:160 the glycosylation of proteins 144:172 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 6 17 theme sialic 853:858 arg1 acids 860:864 sialic acids 853:864 sialic acids 853:864 The presence of sialic acids prolonged the retention of glycopeptides in both chromatographic modes. 28888681 8 18 theme bi-antennary 1172:1183 arg1 glycoform 1185:1193 bi-antennary glycoform 1172:1193 bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1172:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 7 19 theme lower 1051:1055 arg1 sensitivity 1057:1067 a lower sensitivity 1049:1067 a lower sensitivity 1049:1067 The nanoHILIC method provided higher selectivity based on the composition of glycan, compared to nanoRP-LC but a lower sensitivity. 28888681 2 20 theme powerful 381:388 arg1 tools 390:394 the most powerful tools 372:394 the most powerful tools for glycopeptide analysis 372:420 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 8 21 theme outer 1158:1162 arg1 arm 1164:1166 outer arm 1158:1166 outer arm 1158:1166 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 22 theme linkage 1122:1128 arg1 isomers 1130:1136 separate linkage isomers 1113:1136 separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1113:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 2 23 theme tandem 323:328 arg1 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 23 theme tandem 323:328 arg1 spectrometry 335:346 tandem mass spectrometry 323:346 tandem mass spectrometry (RP-LC-MS/MS) 323:360 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 3 24 gly glycoforms 591:600 arg1 peptides 609:616 two peptides 605:616 two peptides obtained from the tryptic digest of hemopexin 605:662 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 8 25 theme glycopeptide 1212:1223 arg1 glycoform 1185:1193 bi-antennary glycoform 1172:1193 bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1172:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 26 gly glycoform 1185:1193 arg1 glycopeptide 1212:1223 SWPAVGDCSSALR glycopeptide 1198:1223 SWPAVGDCSSALR glycopeptide 1198:1223 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 27 gly glycopeptide 1212:1223 arg2 glycopeptide 1212:1223 SWPAVGDCSSALR glycopeptide 1198:1223 SWPAVGDCSSALR glycopeptide 1198:1223 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 4 28 attach attached 745:752 arg1 backbone 774:781 the same peptide backbone 757:781 the same peptide backbone in HILIC 757:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 4 28 attach attached 745:752 arg2 glycans 737:743 glycans 737:743 glycans attached to the same peptide backbone in HILIC 737:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 0 29 theme glycopeptides 117:129 arg1 analysis 95:102 the analysis 91:102 the analysis of hemopexin glycopeptides 91:129 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 2 30 theme chromatography 303:316 arg1 tools 390:394 the most powerful tools 372:394 the most powerful tools for glycopeptide analysis 372:420 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 30 theme chromatography 303:316 arg1 one 365:367 one 365:367 one 365:367 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 30 theme chromatography 303:316 arg1 combination 273:283 A combination 271:283 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) 271:360 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 0 31 theme phase 14:18 arg1 chromatography 63:76 phase versus nano hydrophilic interaction liquid chromatography 14:76 chromatography 63:76 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 0 32 theme hemopexin 107:115 arg1 glycopeptides 117:129 hemopexin glycopeptides 107:129 hemopexin glycopeptides 107:129 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 0 33 from chip 83:86 arg1 analysis 95:102 the analysis 91:102 the analysis of hemopexin glycopeptides 91:129 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 2 34 theme phase 297:301 arg1 chromatography 303:316 reversed phase chromatography 288:316 reversed phase chromatography 288:316 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 0 35 theme hydrophilic 32:42 arg1 chromatography 63:76 phase versus nano hydrophilic interaction liquid chromatography 14:76 chromatography 63:76 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 3 36 theme interaction 485:495 arg1 chromatography 504:517 hydrophilic interaction liquid chromatography 473:517 hydrophilic interaction liquid chromatography (HILIC) 473:525 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 3 36 theme interaction 485:495 arg1 HILIC 520:524 HILIC 520:524 HILIC 520:524 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 8 37 theme nanoHILIC 1074:1082 arg1 able 1095:1098 able 1095:1098 able 1095:1098 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 37 theme nanoHILIC 1074:1082 arg1 method 1084:1089 The nanoHILIC method 1070:1089 The nanoHILIC method 1070:1089 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 1 38 theme orthogonal 203:212 arg1 methods 214:220 orthogonal methods 203:220 orthogonal methods 203:220 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 2 39 theme glycopeptide 400:411 arg1 analysis 413:420 glycopeptide analysis 400:420 glycopeptide analysis 400:420 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 0 40 theme nano 27:30 arg1 chromatography 63:76 phase versus nano hydrophilic interaction liquid chromatography 14:76 chromatography 63:76 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 3 41 theme peptides 609:616 arg1 glycoforms 591:600 separate glycoforms 582:600 separate glycoforms of two peptides obtained from the tryptic digest of hemopexin 582:662 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 0 42 theme liquid 56:61 arg1 chromatography 63:76 phase versus nano hydrophilic interaction liquid chromatography 14:76 chromatography 63:76 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 6 43 theme chromatographic 915:929 arg1 modes 931:935 both chromatographic modes 910:935 both chromatographic modes 910:935 The presence of sialic acids prolonged the retention of glycopeptides in both chromatographic modes. 28888681 4 44 theme glycans 737:743 arg1 polarity 725:732 polarity 725:732 polarity of glycans attached to the same peptide backbone in HILIC 725:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 8 45 from isomers 1130:1136 arg1 glycoform 1185:1193 bi-antennary glycoform 1172:1193 bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1172:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 46 gly fucosylated 1289:1299 arg1 glycoforms 1301:1310 the fucosylated glycoforms 1285:1310 the fucosylated glycoforms 1285:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 3 47 theme hydrophilic 473:483 arg1 chromatography 504:517 hydrophilic interaction liquid chromatography 473:517 hydrophilic interaction liquid chromatography (HILIC) 473:525 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 3 47 theme hydrophilic 473:483 arg1 HILIC 520:524 HILIC 520:524 HILIC 520:524 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 0 48 theme interaction 44:54 arg1 chromatography 63:76 phase versus nano hydrophilic interaction liquid chromatography 14:76 chromatography 63:76 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 0 49 gly glycopeptides 117:129 arg2 glycopeptides 117:129 hemopexin glycopeptides 107:129 hemopexin glycopeptides 107:129 Nano reversed phase versus nano hydrophilic interaction liquid chromatography on a chip in the analysis of hemopexin glycopeptides. 28888681 7 50 theme glycan 1015:1020 arg1 composition 1000:1010 the composition 996:1010 the composition of glycan 996:1020 The nanoHILIC method provided higher selectivity based on the composition of glycan, compared to nanoRP-LC but a lower sensitivity. 28888681 3 51 theme hemopexin 654:662 arg1 digest 644:649 the tryptic digest 632:649 the tryptic digest of hemopexin 632:662 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 4 52 theme peptide 766:772 arg1 backbone 774:781 the same peptide backbone 757:781 the same peptide backbone in HILIC 757:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 4 53 theme time 704:707 arg1 reduction 677:685 reduction 677:685 reduction of the retention time 677:707 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 2 54 with combination 273:283 arg1 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 54 with combination 273:283 arg1 spectrometry 335:346 tandem mass spectrometry 323:346 tandem mass spectrometry (RP-LC-MS/MS) 323:360 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 1 55 gly glycosylation 148:160 arg1 proteins 165:172 proteins 165:172 proteins 165:172 Analysis of the glycosylation of proteins is a challenge that requires orthogonal methods to achieve separation of the diverse glycoforms. 28888681 3 56 theme tryptic 636:642 arg1 digest 644:649 the tryptic digest 632:649 the tryptic digest of hemopexin 632:662 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 4 57 theme same 761:764 arg1 backbone 774:781 the same peptide backbone 757:781 the same peptide backbone in HILIC 757:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 2 58 theme mass 330:333 arg1 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 RP-LC-MS/MS 349:359 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 2 58 theme mass 330:333 arg1 spectrometry 335:346 tandem mass spectrometry 323:346 tandem mass spectrometry (RP-LC-MS/MS) 323:360 A combination of reversed phase chromatography with tandem mass spectrometry (RP-LC-MS/MS) is one of the most powerful tools for glycopeptide analysis. 28888681 8 59 theme glycoforms 1301:1310 arg1 structure 1272:1280 the structure 1268:1280 the structure of the fucosylated glycoforms 1268:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 60 theme structure 1272:1280 arg1 elucidation 1253:1263 the elucidation 1249:1263 the elucidation of the structure of the fucosylated glycoforms 1249:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 8 61 theme separate 1113:1120 arg1 isomers 1130:1136 separate linkage isomers 1113:1136 separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms 1113:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 3 62 theme separate 582:589 arg1 glycoforms 591:600 separate glycoforms 582:600 separate glycoforms of two peptides obtained from the tryptic digest of hemopexin 582:662 In this work, we developed and compared RP-LC and hydrophilic interaction liquid chromatography (HILIC) in nanoscale on a chip combined with MS/MS in order to separate glycoforms of two peptides obtained from the tryptic digest of hemopexin. 28888681 4 63 from backbone 774:781 arg1 HILIC 786:790 HILIC 786:790 HILIC 786:790 We observed reduction of the retention time with decreasing polarity of glycans attached to the same peptide backbone in HILIC. 28888681 8 64 theme SWPAVGDCSSALR 1198:1210 arg1 glycopeptide 1212:1223 SWPAVGDCSSALR glycopeptide 1198:1223 SWPAVGDCSSALR glycopeptide 1198:1223 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 28888681 6 65 theme glycopeptides 893:905 arg1 retention 880:888 the retention 876:888 the retention of glycopeptides 876:905 The presence of sialic acids prolonged the retention of glycopeptides in both chromatographic modes. 28888681 5 66 theme opposite 797:804 arg1 effect 806:811 The opposite effect 793:811 The opposite effect 793:811 The opposite effect was observed for RP-LC. 28888681 8 67 from beneficial 1235:1244 arg1 elucidation 1253:1263 the elucidation 1249:1263 the elucidation of the structure of the fucosylated glycoforms 1249:1310 The nanoHILIC method was able to partially separate linkage isomers of fucose (core and outer arm) on bi-antennary glycoform of SWPAVGDCSSALR glycopeptide, which is beneficial in the elucidation of the structure of the fucosylated glycoforms. 26239923 3 0 theme PMF 285:287 arg1 data 289:292 The PMF data 281:292 The PMF data of protein subunits 281:312 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 4 1 theme complex/hybrid 679:692 arg1 glycans 694:700 sialylated complex/hybrid glycans 668:700 sialylated complex/hybrid glycans 668:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 4 2 gly sialylated 668:677 arg1 glycans 694:700 sialylated complex/hybrid glycans 668:700 sialylated complex/hybrid glycans 668:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 0 3 from characterization 8:23 arg1 colostrum 76:84 buffalo (Bubalus bubalis) colostrum 50:84 buffalo (Bubalus bubalis) colostrum 50:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 7 4 theme effective 1166:1174 arg1 ADCC 1176:1179 effective ADCC 1166:1179 effective ADCC 1166:1179 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 1 5 theme ruminant 137:144 arg1 colostrum 146:154 ruminant colostrum 137:154 ruminant colostrum 137:154 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 0 6 from colostrum 76:84 arg1 characterization 8:23 Glycome characterization 0:23 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 0:85 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 0 6 from colostrum 76:84 arg1 G 43:43 immunoglobulin G 28:43 immunoglobulin G from buffalo (Bubalus bubalis) colostrum 28:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 5 7 link N-linked 742:749 arg1 oligosaccharides 751:766 54 N-linked oligosaccharides 739:766 54 N-linked oligosaccharides 739:766 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 4 8 theme neutral 641:647 arg1 complex/hybrid 649:662 neutral complex/hybrid 641:662 neutral complex/hybrid 641:662 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 4 9 theme chemoselective 561:574 arg1 technique 590:598 chemoselective glycoblotting technique 561:598 chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 561:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 5 10 theme oligosaccharides 751:766 arg1 assignment 725:734 A complete structural assignment 703:734 A complete structural assignment of 54 N-linked oligosaccharides 703:766 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 3 11 theme subunits 305:312 arg1 data 289:292 The PMF data 281:292 The PMF data of protein subunits 281:312 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 1 12 from glycoprotein 121:132 arg1 colostrum 146:154 ruminant colostrum 137:154 ruminant colostrum 137:154 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 3 13 theme CID 484:486 arg1 analysis 494:501 CID MS/MS analysis 484:501 CID MS/MS analysis 484:501 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 7 14 from oligosaccharides 1052:1067 arg1 IgG 1090:1092 buffalo colostrum IgG 1072:1092 buffalo colostrum IgG 1072:1092 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 3 15 theme peptide 389:395 arg1 sequences 397:405 peptide sequences LLIYGATSR and VYNEYLPAPIVR 389:432 peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG 389:479 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 15 theme peptide 389:395 arg1 VYNEYLPAPIVR 421:432 VYNEYLPAPIVR 421:432 VYNEYLPAPIVR 421:432 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 15 theme peptide 389:395 arg1 LLIYGATSR 407:415 LLIYGATSR 407:415 LLIYGATSR 407:415 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 5 16 theme oligosaccharides 808:823 arg1 ratio 792:796 the ratio 788:796 the ratio of sialyl oligosaccharides 788:823 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 5 16 theme oligosaccharides 808:823 arg1 higher 841:846 higher 841:846 higher 841:846 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 6 17 from amount 1012:1017 arg1 present 990:996 present 990:996 present 990:996 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 17 from amount 1012:1017 arg1 fucosylation 885:896 The fucosylation 881:896 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides 881:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 3 18 theme MS/MS 488:492 arg1 analysis 494:501 CID MS/MS analysis 484:501 CID MS/MS analysis 484:501 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 19 theme sequences 397:405 arg1 identification 371:384 the identification 367:384 the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG 367:479 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 6 20 theme diminutive 1001:1010 arg1 amount 1012:1017 diminutive amount 1001:1017 diminutive amount 1001:1017 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 21 located observed 898:905 arg2 fucosylation 885:896 The fucosylation 881:896 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides 881:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 21 located observed 898:905 arg1 oligosaccharides 923:938 more than 20 oligosaccharides 910:938 more than 20 oligosaccharides 910:938 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 21 located observed 898:905 arg1 mannose 946:952 high mannose 941:952 high mannose 941:952 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 21 located observed 898:905 arg1 oligosaccharides 968:983 trisialyl oligosaccharides 958:983 trisialyl oligosaccharides 958:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 21 located observed 898:905 arg2 present 990:996 present 990:996 present 990:996 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 0 22 theme Glycome 0:6 arg1 characterization 8:23 Glycome characterization 0:23 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 0:85 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 4 23 theme N-glycan 508:515 arg1 microheterogeneity 517:534 The N-glycan microheterogeneity 504:534 The N-glycan microheterogeneity 504:534 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 1 24 theme Immunoglobulin 87:100 arg1 glycoprotein 121:132 a major glycoprotein 113:132 a major glycoprotein in ruminant colostrum 113:154 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 1 24 theme Immunoglobulin 87:100 arg1 IgG 105:107 IgG 105:107 IgG 105:107 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 1 24 theme Immunoglobulin 87:100 arg1 G 102:102 Immunoglobulin G 87:102 Immunoglobulin G (IgG) 87:108 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 4 25 theme complex/hybrid 649:662 arg1 identification 609:622 the identification 605:622 the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 605:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 0 26 theme immunoglobulin 28:41 arg1 G 43:43 immunoglobulin G 28:43 immunoglobulin G from buffalo (Bubalus bubalis) colostrum 28:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 7 27 from utilization 1122:1132 arg1 therapies 1146:1154 targeted therapies 1137:1154 targeted therapies 1137:1154 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 6 28 from present 990:996 arg1 amount 1012:1017 diminutive amount 1001:1017 diminutive amount 1001:1017 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 7 29 theme targeted 1137:1144 arg1 therapies 1146:1154 targeted therapies 1137:1154 targeted therapies 1137:1154 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 7 30 from non-fucosyl 1029:1039 arg1 IgG 1090:1092 buffalo colostrum IgG 1072:1092 buffalo colostrum IgG 1072:1092 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 5 31 theme complete 705:712 arg1 assignment 725:734 A complete structural assignment 703:734 A complete structural assignment of 54 N-linked oligosaccharides 703:766 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 2 32 theme day 163:165 arg1 protein 185:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein was purified on Sephadex G-100 and its mass was determined by MALDI-TOF as 147.848 KDa. 26239923 4 33 theme glycans 694:700 arg1 identification 609:622 the identification 605:622 the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 605:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 3 34 theme IgG 477:479 arg1 light 451:455 light 451:455 light 451:455 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 34 theme IgG 477:479 arg1 chains 467:472 heavy chains 461:472 heavy chains 461:472 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 7 35 theme anti-inflammatory 1185:1201 arg1 responses 1203:1211 anti-inflammatory responses 1185:1211 anti-inflammatory responses 1185:1211 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 5 36 theme structural 714:723 arg1 assignment 725:734 A complete structural assignment 703:734 A complete structural assignment of 54 N-linked oligosaccharides 703:766 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 2 37 theme First 157:161 arg1 protein 185:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein was purified on Sephadex G-100 and its mass was determined by MALDI-TOF as 147.848 KDa. 26239923 4 38 theme mannose 632:638 arg1 identification 609:622 the identification 605:622 the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 605:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 0 39 theme G 43:43 arg1 characterization 8:23 Glycome characterization 0:23 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 0:85 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 7 40 theme ample 1102:1106 arg1 scope 1108:1112 ample scope 1102:1112 ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses 1102:1211 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 5 41 theme neutral 860:866 arg1 saccharides 868:878 neutral saccharides 860:878 neutral saccharides 860:878 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 4 42 theme high 627:630 arg1 mannose 632:638 high mannose 627:638 high mannose 627:638 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 5 43 theme sialyl 801:806 arg1 oligosaccharides 808:823 sialyl oligosaccharides 801:823 sialyl oligosaccharides 801:823 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 6 44 theme trisialyl 958:966 arg1 oligosaccharides 968:983 trisialyl oligosaccharides 958:983 trisialyl oligosaccharides 958:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 1 45 gly glycoprotein 121:132 arg1 glycoprotein 121:132 a major glycoprotein 113:132 a major glycoprotein in ruminant colostrum 113:154 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 1 45 gly glycoprotein 121:132 arg1 G 102:102 Immunoglobulin G 87:102 Immunoglobulin G (IgG) 87:108 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 0 46 theme Bubalus 59:65 arg1 colostrum 76:84 buffalo (Bubalus bubalis) colostrum 50:84 buffalo (Bubalus bubalis) colostrum 50:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 4 47 with technique 590:598 arg1 identification 609:622 the identification 605:622 the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 605:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 0 48 theme buffalo 50:56 arg1 colostrum 76:84 buffalo (Bubalus bubalis) colostrum 50:84 buffalo (Bubalus bubalis) colostrum 50:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 3 49 dep sequences 397:405 arg1 sequences 397:405 peptide sequences LLIYGATSR and VYNEYLPAPIVR 389:432 peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG 389:479 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 49 dep sequences 397:405 arg1 VYNEYLPAPIVR 421:432 VYNEYLPAPIVR 421:432 VYNEYLPAPIVR 421:432 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 49 dep sequences 397:405 arg1 LLIYGATSR 407:415 LLIYGATSR 407:415 LLIYGATSR 407:415 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 3 50 theme heavy 461:465 arg1 chains 467:472 heavy chains 461:472 heavy chains 461:472 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 6 51 located present 990:996 arg2 fucosylation 885:896 The fucosylation 881:896 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides 881:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 51 located present 990:996 arg1 amount 1012:1017 diminutive amount 1001:1017 diminutive amount 1001:1017 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 51 located present 990:996 arg2 present 990:996 present 990:996 present 990:996 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 5 52 theme N-linked 742:749 arg1 oligosaccharides 751:766 54 N-linked oligosaccharides 739:766 54 N-linked oligosaccharides 739:766 A complete structural assignment of 54 N-linked oligosaccharides were identified and the ratio of sialyl oligosaccharides was found to be higher compared to neutral saccharides. 26239923 7 53 theme buffalo 1072:1078 arg1 IgG 1090:1092 buffalo colostrum IgG 1072:1092 buffalo colostrum IgG 1072:1092 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 2 54 theme colostrum 175:183 arg1 protein 185:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein was purified on Sephadex G-100 and its mass was determined by MALDI-TOF as 147.848 KDa. 26239923 7 55 theme sialyl 1045:1050 arg1 oligosaccharides 1052:1067 The high non-fucosyl and sialyl oligosaccharides 1020:1067 oligosaccharides 1052:1067 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 4 56 theme glycoblotting 576:588 arg1 technique 590:598 chemoselective glycoblotting technique 561:598 chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans 561:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 26239923 0 57 theme bubalis 67:73 arg1 colostrum 76:84 buffalo (Bubalus bubalis) colostrum 50:84 buffalo (Bubalus bubalis) colostrum 50:84 Glycome characterization of immunoglobulin G from buffalo (Bubalus bubalis) colostrum. 26239923 2 58 theme buffalo 167:173 arg1 protein 185:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein 157:191 First day buffalo colostrum protein was purified on Sephadex G-100 and its mass was determined by MALDI-TOF as 147.848 KDa. 26239923 7 59 theme colostrum 1080:1088 arg1 IgG 1090:1092 buffalo colostrum IgG 1072:1092 buffalo colostrum IgG 1072:1092 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 7 60 theme high 1024:1027 arg1 non-fucosyl 1029:1039 The high non-fucosyl and sialyl oligosaccharides 1020:1067 non-fucosyl 1029:1039 The high non-fucosyl and sialyl oligosaccharides in buffalo colostrum IgG provide ample scope for its utilization in targeted therapies to elicit effective ADCC and anti-inflammatory responses. 26239923 1 61 theme major 115:119 arg1 glycoprotein 121:132 a major glycoprotein 113:132 a major glycoprotein in ruminant colostrum 113:154 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 1 61 theme major 115:119 arg1 G 102:102 Immunoglobulin G 87:102 Immunoglobulin G (IgG) 87:108 Immunoglobulin G (IgG) is a major glycoprotein in ruminant colostrum. 26239923 3 62 theme protein 297:303 arg1 subunits 305:312 protein subunits 297:312 protein subunits 297:312 The PMF data of protein subunits revealed its homology to IgG, which was supported by the identification of peptide sequences LLIYGATSR and VYNEYLPAPIVR corresponding to light and heavy chains of IgG by CID MS/MS analysis. 26239923 6 63 theme high 941:944 arg1 mannose 946:952 high mannose 941:952 high mannose 941:952 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 64 attach present 990:996 arg2 fucosylation 885:896 The fucosylation 881:896 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides 881:983 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 64 attach present 990:996 arg1 amount 1012:1017 diminutive amount 1001:1017 diminutive amount 1001:1017 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 6 64 attach present 990:996 arg2 present 990:996 present 990:996 present 990:996 The fucosylation observed in more than 20 oligosaccharides, high mannose and trisialyl oligosaccharides were present in diminutive amount. 26239923 4 65 theme sialylated 668:677 arg1 glycans 694:700 sialylated complex/hybrid glycans 668:700 sialylated complex/hybrid glycans 668:700 The N-glycan microheterogeneity was established based on chemoselective glycoblotting technique with the identification of high mannose, neutral complex/hybrid and sialylated complex/hybrid glycans. 24612865 1 0 theme β-linked 374:381 arg1 moiety 399:404 a β-linked glucuronic acid moiety 372:404 a β-linked glucuronic acid moiety 372:404 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 5 1 theme disaccharides 1214:1226 arg1 yields 1204:1209 good yields 1199:1209 good yields of disaccharides with high β-selectivity 1199:1250 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 3 2 theme subsequent 810:819 arg1 introduction 821:832 subsequent introduction 810:832 subsequent introduction 810:832 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 1 3 theme GXM 281:283 arg1 polysaccharide 294:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 4 theme building 67:74 arg1 blocks 76:81 building blocks 67:81 building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures 67:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 1 5 theme glucuronic 383:392 arg1 moiety 399:404 a β-linked glucuronic acid moiety 372:404 a β-linked glucuronic acid moiety 372:404 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 6 dep neoformans 116:125 arg1 structures 151:160 capsular polysaccharide structures 127:160 Cryptococcus neoformans capsular polysaccharide structures 103:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 5 7 from glycosylations 1026:1039 arg1 presence 1128:1135 the presence 1124:1135 the presence of nitrile solvents 1124:1155 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 1 8 theme capsular 285:292 arg1 polysaccharide 294:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 1 9 theme acid 394:397 arg1 moiety 399:404 a β-linked glucuronic acid moiety 372:404 a β-linked glucuronic acid moiety 372:404 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 2 10 theme acetyl 637:642 arg1 group 644:648 a primary acetyl group 627:648 a primary acetyl group 627:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 6 11 theme glucuronyl 1295:1304 arg1 donors 1320:1325 recently reported glucuronyl thioglycoside donors 1277:1325 recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal 1277:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 2 12 theme acid 569:572 arg1 residue 574:580 a β-linked glucuronic acid residue 547:580 a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group 547:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 5 13 theme good 1199:1202 arg1 yields 1204:1209 good yields 1199:1209 good yields of disaccharides with high β-selectivity 1199:1250 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 2 14 theme primary 629:635 arg1 group 644:648 a primary acetyl group 627:648 a primary acetyl group 627:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 6 15 theme reported 1286:1293 arg1 donors 1320:1325 recently reported glucuronyl thioglycoside donors 1277:1325 recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal 1277:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 0 16 theme Cryptococcus 103:114 arg1 neoformans 116:125 Cryptococcus neoformans 103:125 Cryptococcus neoformans capsular polysaccharide structures 103:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 2 17 theme β-linked 549:556 arg1 residue 574:580 a β-linked glucuronic acid residue 547:580 a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group 547:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 3 18 theme acetyl 867:872 arg1 group 874:878 the acetyl group 863:878 the acetyl group 863:878 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 3 19 dep developed 682:690 arg1 using 697:701 using 697:701 using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group 697:878 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 2 20 theme glucuronic 558:567 arg1 residue 574:580 a β-linked glucuronic acid residue 547:580 a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group 547:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 1 21 theme 6-O-acetyl 412:421 arg1 group 423:427 a 6-O-acetyl group 410:427 a 6-O-acetyl group 410:427 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 3 22 theme group 874:878 arg1 benzyls 798:804 benzyls 798:804 benzyls 798:804 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 3 22 theme group 874:878 arg1 introduction 821:832 subsequent introduction 810:832 subsequent introduction 810:832 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 6 23 theme donors 1320:1325 arg1 use 1270:1272 the use 1266:1272 the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal 1266:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 4 24 theme second 885:890 arg1 route 892:896 The second route 881:896 The second route 881:896 The second route explored the possibility to achieve β-selectivity using glucuronyl donors without acyl protecting groups. 24612865 5 25 theme nitrile 1140:1146 arg1 solvents 1148:1155 nitrile solvents 1140:1155 nitrile solvents 1140:1155 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 0 26 theme protected 20:28 arg1 -α-d-Man 44:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 1 27 theme easy 313:316 arg1 access 318:323 an easy access 310:323 an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group 310:427 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 28 theme capsular 127:134 arg1 structures 151:160 capsular polysaccharide structures 127:160 Cryptococcus neoformans capsular polysaccharide structures 103:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 5 29 theme BF3-etherate 1004:1015 arg1 glycosylations 1026:1039 BF3-etherate promoted glycosylations 1004:1039 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures 1004:1179 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 1 30 theme large 203:207 arg1 structures 225:234 large oligosaccharide structures 203:234 large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide 203:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 31 theme benzyl 13:18 arg1 -α-d-Man 44:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 5 32 theme high 1233:1236 arg1 β-selectivity 1238:1250 high β-selectivity 1233:1250 high β-selectivity 1233:1250 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 0 33 theme neoformans 116:125 arg1 construction 87:98 construction 87:98 construction of Cryptococcus neoformans capsular polysaccharide structures 87:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 4 34 theme acyl 980:983 arg1 groups 996:1001 acyl protecting groups 980:1001 acyl protecting groups 980:1001 The second route explored the possibility to achieve β-selectivity using glucuronyl donors without acyl protecting groups. 24612865 5 35 theme promoted 1017:1024 arg1 glycosylations 1026:1039 BF3-etherate promoted glycosylations 1004:1039 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures 1004:1179 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 1 36 theme oligosaccharide 209:223 arg1 structures 225:234 large oligosaccharide structures 203:234 large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide 203:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 5 37 with glycosylations 1026:1039 arg1 trichloroacetimidate 1100:1119 benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate 1046:1119 benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate 1046:1119 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 4 38 theme protecting 985:994 arg1 groups 996:1001 acyl protecting groups 980:1001 acyl protecting groups 980:1001 The second route explored the possibility to achieve β-selectivity using glucuronyl donors without acyl protecting groups. 24612865 1 39 link β-linked 374:381 arg1 moiety 399:404 a β-linked glucuronic acid moiety 372:404 a β-linked glucuronic acid moiety 372:404 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 1 40 theme structures 225:234 arg1 synthesis 190:198 the synthesis 186:198 the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide 186:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 41 theme β-D-GlcA- 30:38 arg1 -α-d-Man 44:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 1 42 theme thiodisaccharide 328:343 arg1 blocks 354:359 thiodisaccharide building blocks 328:359 thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group 328:427 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 43 theme polysaccharide 136:149 arg1 structures 151:160 capsular polysaccharide structures 127:160 Cryptococcus neoformans capsular polysaccharide structures 103:160 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 6 44 theme building 1467:1474 arg1 blocks 1476:1481 desired disaccharide building blocks 1446:1481 desired disaccharide building blocks 1446:1481 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 6 45 theme 2,4-silylene 1351:1362 arg1 acetal 1364:1369 a cyclic 2,4-silylene acetal 1342:1369 a cyclic 2,4-silylene acetal 1342:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 5 46 from temperatures 1168:1179 arg1 glycosylations 1026:1039 BF3-etherate promoted glycosylations 1004:1039 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures 1004:1179 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 2 47 theme Several 443:449 arg1 pathways 451:458 Several pathways 443:458 Several pathways to such building blocks 443:482 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 5 48 theme low 1164:1166 arg1 temperatures 1168:1179 low temperatures 1164:1179 low temperatures 1164:1179 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 1 49 theme building 345:352 arg1 blocks 354:359 thiodisaccharide building blocks 328:359 thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group 328:427 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 6 50 theme desired 1446:1452 arg1 blocks 1476:1481 desired disaccharide building blocks 1446:1481 desired disaccharide building blocks 1446:1481 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 5 51 gly glycosylations 1026:1039 arg2 temperatures 1168:1179 low temperatures 1164:1179 low temperatures 1164:1179 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 6 52 theme cyclic 1344:1349 arg1 acetal 1364:1369 a cyclic 2,4-silylene acetal 1342:1369 a cyclic 2,4-silylene acetal 1342:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 0 53 theme 1→2 40:42 arg1 -α-d-Man 44:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 benzyl protected β-D-GlcA-(1→2)-α-d-Man 13:51 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 3 54 theme glucosyl 715:722 arg1 donors 724:729 benzoylated glucosyl donors 703:729 benzoylated glucosyl donors 703:729 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 6 55 theme efficient 1402:1410 arg1 way 1439:1441 another efficient and completely β-selective way 1394:1441 another efficient and completely β-selective way to desired disaccharide building blocks 1394:1481 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 3 56 theme efficient 655:663 arg1 routes 665:670 Two efficient routes 651:670 Two efficient routes 651:670 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 4 57 theme glucuronyl 954:963 arg1 donors 965:970 glucuronyl donors 954:970 glucuronyl donors 954:970 The second route explored the possibility to achieve β-selectivity using glucuronyl donors without acyl protecting groups. 24612865 5 58 theme solvents 1148:1155 arg1 presence 1128:1135 the presence 1124:1135 the presence of nitrile solvents 1124:1155 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 6 59 theme thioglycoside 1306:1318 arg1 donors 1320:1325 recently reported glucuronyl thioglycoside donors 1277:1325 recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal 1277:1369 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 6 60 theme disaccharide 1454:1465 arg1 blocks 1476:1481 desired disaccharide building blocks 1446:1481 desired disaccharide building blocks 1446:1481 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 5 61 with disaccharides 1214:1226 arg1 β-selectivity 1238:1250 high β-selectivity 1233:1250 high β-selectivity 1233:1250 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 3 62 theme carboxyl 841:848 arg1 function 850:857 the carboxyl function 837:857 the carboxyl function 837:857 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 6 63 theme β-selective 1427:1437 arg1 way 1439:1441 another efficient and completely β-selective way 1394:1441 another efficient and completely β-selective way to desired disaccharide building blocks 1394:1481 Furthermore, the use of recently reported glucuronyl thioglycoside donors protected with a cyclic 2,4-silylene acetal was found to represent another efficient and completely β-selective way to desired disaccharide building blocks. 24612865 2 64 theme such 463:466 arg1 blocks 477:482 such building blocks 463:482 such building blocks 463:482 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 3 65 theme function 850:857 arg1 benzyls 798:804 benzyls 798:804 benzyls 798:804 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 3 65 theme function 850:857 arg1 introduction 821:832 subsequent introduction 810:832 subsequent introduction 810:832 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 3 66 theme benzoylated 703:713 arg1 donors 724:729 benzoylated glucosyl donors 703:729 benzoylated glucosyl donors 703:729 Two efficient routes have been developed, one using benzoylated glucosyl donors to form the β-linkage followed by a change of protecting groups to benzyls and subsequent introduction of the carboxyl function and the acetyl group. 24612865 5 67 theme uronate 1092:1098 arg1 trichloroacetimidate 1100:1119 benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate 1046:1119 benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate 1046:1119 BF3-etherate promoted glycosylations with benzyl (2,3,4-tri-O-benzyl-α-D-glucupyranosyl)uronate trichloroacetimidate in the presence of nitrile solvents and at low temperatures reproducibly gave good yields of disaccharides with high β-selectivity. 24612865 2 68 link β-linked 549:556 arg1 residue 574:580 a β-linked glucuronic acid residue 547:580 a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group 547:648 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 1 69 theme Cryptococcus 257:268 arg1 neoformans 270:279 Cryptococcus neoformans 257:279 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 24612865 0 70 theme -α-d-Man 44:51 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man 0:51 Synthesis of benzyl protected β-D-GlcA-(1→2)-α-d-Man thioglycoside building blocks for construction of Cryptococcus neoformans capsular polysaccharide structures. 24612865 2 71 theme building 468:475 arg1 blocks 477:482 such building blocks 463:482 such building blocks 463:482 Several pathways to such building blocks have been investigated, addressing the problem of constructing a β-linked glucuronic acid residue protected with groups that are orthogonal to a primary acetyl group. 24612865 1 72 theme neoformans 270:279 arg1 polysaccharide 294:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 the Cryptococcus neoformans GXM capsular polysaccharide 253:307 In a project targeting the synthesis of large oligosaccharide structures corresponding to the Cryptococcus neoformans GXM capsular polysaccharide, an easy access to thiodisaccharide building blocks comprising a β-linked glucuronic acid moiety and a 6-O-acetyl group was required. 28084659 5 0 theme products 831:838 arg1 cleavage 808:815 Light-induced cleavage 794:815 Light-induced cleavage of the glycan products from the solid support followed by global deprotection 794:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 6 1 theme oligosaccharides 997:1012 arg1 incubation 975:984 incubation 975:984 incubation of the MLG oligosaccharides with lichenase 975:1027 After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC-MS. 28084659 7 2 theme lichenases 1243:1252 arg1 specificity 1228:1238 the substrate specificity 1214:1238 the substrate specificity of lichenases 1214:1252 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 7 3 theme active 1143:1148 arg1 site 1150:1153 the enzyme's active site 1130:1153 the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered 1130:1275 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 4 4 theme C-3 676:678 arg1 position 687:694 the C-3 or C-4 position 672:694 position 687:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 7 5 theme substrate 1218:1226 arg1 specificity 1228:1238 the substrate specificity 1214:1238 the substrate specificity of lichenases 1214:1252 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 3 6 theme substrate 492:500 arg1 specificities 502:514 the substrate specificities 488:514 the substrate specificities of Bacillus subtilis lichenase 488:545 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 1 7 theme important 233:241 arg1 tool 255:258 an important biochemical tool 230:258 an important biochemical tool for the structural characterization of MLGs 230:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 7 theme important 233:241 arg1 lichenase 217:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 6 8 with incubation 975:984 arg1 lichenase 1019:1027 lichenase 1019:1027 lichenase 1019:1027 After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC-MS. 28084659 3 9 theme lichenase 537:545 arg1 specificities 502:514 the substrate specificities 488:514 the substrate specificities of Bacillus subtilis lichenase 488:545 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 4 10 from group 663:667 arg1 position 687:694 the C-3 or C-4 position 672:694 position 687:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 1 11 theme biochemical 243:253 arg1 tool 255:258 an important biochemical tool 230:258 an important biochemical tool for the structural characterization of MLGs 230:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 11 theme biochemical 243:253 arg1 lichenase 217:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 4 12 theme temporary 599:607 arg1 group 663:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group 597:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position 597:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 4 13 theme automated 755:763 arg1 synthesizer 781:791 an automated oligosaccharide synthesizer 752:791 an automated oligosaccharide synthesizer 752:791 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 3 14 theme Several 393:399 arg1 oligosaccharides 413:428 Several defined MLG oligosaccharides 393:428 Several defined MLG oligosaccharides obtained by automated glycan assembly 393:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 3 15 theme MLG 409:411 arg1 oligosaccharides 413:428 Several defined MLG oligosaccharides 393:428 Several defined MLG oligosaccharides obtained by automated glycan assembly 393:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 0 16 theme Substrate 108:116 arg1 Specificity 118:128 the Substrate Specificity 104:128 the Substrate Specificity of Lichenase 104:141 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 5 17 theme MLG 910:912 arg1 oligosaccharides 914:929 seven MLG oligosaccharides 904:929 seven MLG oligosaccharides of different length and connectivity 904:966 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 3 18 theme Bacillus 519:526 arg1 lichenase 537:545 Bacillus subtilis lichenase 519:545 Bacillus subtilis lichenase 519:545 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 2 19 theme feed 363:366 arg1 industries 381:390 the brewery, animal feed, and biofuel industries 343:390 the brewery, animal feed, and biofuel industries 343:390 It holds potential for application in the brewery, animal feed, and biofuel industries. 28084659 0 20 theme Glucan 14:19 arg1 Oligosaccharides 21:36 Mixed-Linkage Glucan Oligosaccharides 0:36 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly 0:74 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 0 20 theme Glucan 14:19 arg1 Tools 85:89 Tools 85:89 Tools 85:89 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 4 21 theme glucose 552:558 arg1 BBs 577:579 BBs 577:579 BBs 577:579 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 4 21 theme glucose 552:558 arg1 blocks 569:574 Two glucose building blocks 548:574 Two glucose building blocks (BBs) 548:580 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 2 22 from application 328:338 arg1 industries 381:390 the brewery, animal feed, and biofuel industries 343:390 the brewery, animal feed, and biofuel industries 343:390 It holds potential for application in the brewery, animal feed, and biofuel industries. 28084659 2 23 theme animal 356:361 arg1 feed 363:366 animal feed 356:366 animal feed 356:366 It holds potential for application in the brewery, animal feed, and biofuel industries. 28084659 0 24 theme Mixed-Linkage 0:12 arg1 Oligosaccharides 21:36 Mixed-Linkage Glucan Oligosaccharides 0:36 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly 0:74 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 0 24 theme Mixed-Linkage 0:12 arg1 Tools 85:89 Tools 85:89 Tools 85:89 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 6 25 theme MLG 993:995 arg1 oligosaccharides 997:1012 the MLG oligosaccharides 989:1012 the MLG oligosaccharides 989:1012 After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC-MS. 28084659 5 26 theme connectivity 955:966 arg1 oligosaccharides 914:929 seven MLG oligosaccharides 904:929 seven MLG oligosaccharides of different length and connectivity 904:966 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 3 27 theme defined 401:407 arg1 oligosaccharides 413:428 Several defined MLG oligosaccharides 393:428 Several defined MLG oligosaccharides obtained by automated glycan assembly 393:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 7 28 theme other 1176:1180 arg1 evidence 1189:1196 other recent evidence 1176:1196 other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered 1176:1275 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 8 29 theme useful 1344:1349 arg1 oligosaccharides 1323:1338 synthetic MLG oligosaccharides 1309:1338 synthetic MLG oligosaccharides 1309:1338 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 8 29 theme useful 1344:1349 arg1 tools 1351:1355 useful tools 1344:1355 useful tools to analyze mixed-linkage β-glucanases 1344:1393 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 0 30 theme Lichenase 133:141 arg1 Specificity 118:128 the Substrate Specificity 104:128 the Substrate Specificity of Lichenase 104:141 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 1 31 dep mixed-linkage 148:160 arg1 specific 189:196 specific 189:196 specific 189:196 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 31 dep mixed-linkage 148:160 arg1 1→3 163:165 1→3 163:165 1→3 163:165 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 3 32 theme automated 442:450 arg1 assembly 459:466 automated glycan assembly 442:466 automated glycan assembly 442:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 5 33 theme glycan 824:829 arg1 products 831:838 the glycan products 820:838 the glycan products from the solid support followed by global deprotection 820:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 4 34 theme building 560:567 arg1 BBs 577:579 BBs 577:579 BBs 577:579 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 4 34 theme building 560:567 arg1 blocks 569:574 Two glucose building blocks 548:574 Two glucose building blocks (BBs) 548:580 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 7 35 with line 1166:1169 arg1 evidence 1189:1196 other recent evidence 1176:1196 other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered 1176:1275 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 4 36 theme different 716:724 arg1 oligosaccharides 726:741 different oligosaccharides 716:741 different oligosaccharides 716:741 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 3 37 theme glycan 452:457 arg1 assembly 459:466 automated glycan assembly 442:466 automated glycan assembly 442:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 0 38 theme Automated 50:58 arg1 Assembly 67:74 Automated Glycan Assembly 50:74 Automated Glycan Assembly 50:74 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 1 39 theme structural 268:277 arg1 characterization 279:294 the structural characterization 264:294 the structural characterization of MLGs 264:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 8 40 theme synthetic 1309:1317 arg1 oligosaccharides 1323:1338 synthetic MLG oligosaccharides 1309:1338 synthetic MLG oligosaccharides 1309:1338 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 8 40 theme synthetic 1309:1317 arg1 tools 1351:1355 useful tools 1344:1355 useful tools to analyze mixed-linkage β-glucanases 1344:1393 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 4 41 theme protecting 652:661 arg1 group 663:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group 597:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position 597:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 2 42 theme biofuel 373:379 arg1 industries 381:390 the brewery, animal feed, and biofuel industries 343:390 the brewery, animal feed, and biofuel industries 343:390 It holds potential for application in the brewery, animal feed, and biofuel industries. 28084659 7 43 theme digestion 1085:1093 arg1 experiments 1095:1105 These digestion experiments 1079:1105 These digestion experiments 1079:1105 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 6 44 theme digestion 1034:1042 arg1 products 1044:1051 the digestion products 1030:1051 the digestion products 1030:1051 After incubation of the MLG oligosaccharides with lichenase, the digestion products were analyzed by HPLC-MS. 28084659 4 45 theme chloride 636:643 arg1 group 663:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group 597:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position 597:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 5 46 theme Light-induced 794:806 arg1 cleavage 808:815 Light-induced cleavage 794:815 Light-induced cleavage of the glycan products from the solid support followed by global deprotection 794:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 5 47 from cleavage 808:815 arg1 support 855:861 the solid support 845:861 the solid support followed by global deprotection 845:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 0 48 theme Glycan 60:65 arg1 Assembly 67:74 Automated Glycan Assembly 50:74 Automated Glycan Assembly 50:74 Mixed-Linkage Glucan Oligosaccharides Produced by Automated Glycan Assembly Serve as Tools To Determine the Substrate Specificity of Lichenase. 28084659 1 49 theme MLGs 299:302 arg1 characterization 279:294 the structural characterization 264:294 the structural characterization of MLGs 264:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 3 50 used used 472:475 arg2 oligosaccharides 413:428 Several defined MLG oligosaccharides 393:428 Several defined MLG oligosaccharides obtained by automated glycan assembly 393:466 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 3 51 dep Bacillus 519:526 arg1 subtilis 528:535 subtilis 528:535 subtilis 528:535 Several defined MLG oligosaccharides obtained by automated glycan assembly are used to analyze the substrate specificities of Bacillus subtilis lichenase. 28084659 1 52 theme mixed-linkage 148:160 arg1 tool 255:258 an important biochemical tool 230:258 an important biochemical tool for the structural characterization of MLGs 230:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 52 theme mixed-linkage 148:160 arg1 lichenase 217:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 5 53 theme different 934:942 arg1 length 944:949 different length 934:949 different length 934:949 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 5 54 theme global 875:880 arg1 deprotection 882:893 global deprotection 875:893 global deprotection 875:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 4 55 theme oligosaccharide 765:779 arg1 synthesizer 781:791 an automated oligosaccharide synthesizer 752:791 an automated oligosaccharide synthesizer 752:791 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 5 56 theme length 944:949 arg1 oligosaccharides 914:929 seven MLG oligosaccharides 904:929 seven MLG oligosaccharides of different length and connectivity 904:966 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 8 57 theme mixed-linkage 1368:1380 arg1 β-glucanases 1382:1393 mixed-linkage β-glucanases 1368:1393 mixed-linkage β-glucanases 1368:1393 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 1 58 theme glycosyl 198:205 arg1 tool 255:258 an important biochemical tool 230:258 an important biochemical tool for the structural characterization of MLGs 230:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 58 theme glycosyl 198:205 arg1 lichenase 217:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 5 59 theme solid 849:853 arg1 support 855:861 the solid support 845:861 the solid support followed by global deprotection 845:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 4 60 theme fluorenylmethyloxycarbonyl 609:634 arg1 group 663:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group 597:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position 597:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 2 61 theme brewery 347:353 arg1 industries 381:390 the brewery, animal feed, and biofuel industries 343:390 the brewery, animal feed, and biofuel industries 343:390 It holds potential for application in the brewery, animal feed, and biofuel industries. 28084659 4 62 theme Fmoc 646:649 arg1 group 663:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group 597:667 a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position 597:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 28084659 1 63 theme hydrolase 207:215 arg1 tool 255:258 an important biochemical tool 230:258 an important biochemical tool for the structural characterization of MLGs 230:302 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 1 63 theme hydrolase 207:215 arg1 lichenase 217:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase 144:225 The mixed-linkage (1→3),(1→4)-d-glucan (MLG) specific glycosyl hydrolase lichenase is an important biochemical tool for the structural characterization of MLGs. 28084659 7 64 theme recent 1182:1187 arg1 evidence 1189:1196 other recent evidence 1176:1196 other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered 1176:1275 These digestion experiments provided insights into the enzyme's active site that is in line with other recent evidence suggesting that the substrate specificity of lichenases has to be reconsidered. 28084659 5 65 from support 855:861 arg1 products 831:838 the glycan products 820:838 the glycan products from the solid support followed by global deprotection 820:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 5 65 from support 855:861 arg1 cleavage 808:815 Light-induced cleavage 794:815 Light-induced cleavage of the glycan products from the solid support followed by global deprotection 794:893 Light-induced cleavage of the glycan products from the solid support followed by global deprotection provided seven MLG oligosaccharides of different length and connectivity. 28084659 8 66 theme MLG 1319:1321 arg1 oligosaccharides 1323:1338 synthetic MLG oligosaccharides 1309:1338 synthetic MLG oligosaccharides 1309:1338 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 8 66 theme MLG 1319:1321 arg1 tools 1351:1355 useful tools 1344:1355 useful tools to analyze mixed-linkage β-glucanases 1344:1393 These results demonstrate that synthetic MLG oligosaccharides are useful tools to analyze mixed-linkage β-glucanases. 28084659 4 67 theme C-4 683:685 arg1 position 687:694 the C-3 or C-4 position 672:694 position 687:694 Two glucose building blocks (BBs), equipped with a temporary fluorenylmethyloxycarbonyl chloride (Fmoc) protecting group in the C-3 or C-4 position, served to assemble different oligosaccharides by using an automated oligosaccharide synthesizer. 26216755 2 0 theme Gly77 441:445 arg1 mutation 429:436 mutation 429:436 mutation of Gly77 to tryptophan (CrChiA-G77W) 429:473 The removal from subsite +3 through mutation of Trp168 to alanine suppressed TG activity, while introduction into subsite +1 through mutation of Gly77 to tryptophan (CrChiA-G77W) enhanced TG activity. 26216755 4 1 theme key 895:897 arg1 residues 854:861 Aromatic residues 845:861 Aromatic residues in the aglycon-binding site 845:889 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 4 1 theme key 895:897 arg1 determinants 899:910 key determinants 895:910 key determinants of TG activity of GH18 chitinases 895:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 1 2 from activity 153:160 arg1 revoluta 211:218 Cycas revoluta 205:218 Cycas revoluta 205:218 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 1 2 from activity 153:160 arg1 CrChiA 222:227 CrChiA 222:227 CrChiA 222:227 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 1 2 from activity 153:160 arg1 cycad 198:202 the cycad 194:202 the cycad 194:202 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 3 3 theme GlcNAc 822:827 arg1 absence 811:817 the absence 807:817 the absence of GlcNAc at the subsite 807:842 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 4 4 from residues 854:861 arg1 site 886:889 the aglycon-binding site 866:889 the aglycon-binding site 866:889 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 0 5 theme tryptophan 106:115 arg1 chain 122:126 a tryptophan side chain 104:126 a tryptophan side chain 104:126 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 4 6 theme activity 918:925 arg1 residues 854:861 Aromatic residues 845:861 Aromatic residues in the aglycon-binding site 845:889 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 4 6 theme activity 918:925 arg1 determinants 899:910 key determinants 895:910 key determinants of TG activity of GH18 chitinases 895:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 2 7 from subsite 313:319 arg1 removal 300:306 The removal 296:306 The removal from subsite +3 through mutation of Trp168 to alanine 296:360 The removal from subsite +3 through mutation of Trp168 to alanine suppressed TG activity, while introduction into subsite +1 through mutation of Gly77 to tryptophan (CrChiA-G77W) enhanced TG activity. 26216755 4 8 theme GH18 930:933 arg1 chitinases 935:944 GH18 chitinases 930:944 GH18 chitinases 930:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 2 9 theme Trp168 344:349 arg1 mutation 332:339 mutation 332:339 mutation of Trp168 to alanine 332:360 The removal from subsite +3 through mutation of Trp168 to alanine suppressed TG activity, while introduction into subsite +1 through mutation of Gly77 to tryptophan (CrChiA-G77W) enhanced TG activity. 26216755 4 10 theme chitinases 935:944 arg1 activity 918:925 TG activity 915:925 TG activity of GH18 chitinases 915:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 1 11 theme family 167:172 arg1 chitinase 179:187 a family GH18 chitinase 165:187 a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 165:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 3 12 with structures 509:518 arg1 residues 615:622 one or two N-acetylglucosamine residues 584:622 one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively 584:667 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 3 13 theme side 694:697 arg1 chain 699:703 the Trp77 side chain 684:703 the Trp77 side chain 684:703 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 1 14 theme GH18 174:177 arg1 chitinase 179:187 a family GH18 chitinase 165:187 a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 165:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 0 15 theme transglycosylation 18:35 arg1 activity 37:44 the transglycosylation activity 14:44 the transglycosylation activity of plant family GH18 chitinase 14:75 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 3 16 theme crystal 501:507 arg1 structures 509:518 The crystal structures 497:518 The crystal structures 497:518 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 1 17 theme chitinase 179:187 arg1 activity 153:160 Transglycosylation (TG) activity 129:160 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 129:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 0 18 theme side 117:120 arg1 chain 122:126 a tryptophan side chain 104:126 a tryptophan side chain 104:126 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 2 19 theme TG 373:374 arg1 activity 376:383 TG activity 373:383 TG activity 373:383 The removal from subsite +3 through mutation of Trp168 to alanine suppressed TG activity, while introduction into subsite +1 through mutation of Gly77 to tryptophan (CrChiA-G77W) enhanced TG activity. 26216755 4 20 theme TG 915:916 arg1 activity 918:925 TG activity 915:925 TG activity of GH18 chitinases 915:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 3 21 theme Trp77 688:692 arg1 chain 699:703 the Trp77 side chain 684:703 the Trp77 side chain 684:703 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 4 22 theme Aromatic 845:852 arg1 residues 854:861 Aromatic residues 845:861 Aromatic residues in the aglycon-binding site 845:889 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 4 22 theme Aromatic 845:852 arg1 determinants 899:910 key determinants 895:910 key determinants of TG activity of GH18 chitinases 895:944 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 3 23 theme mutant 542:547 arg1 structures 509:518 The crystal structures 497:518 The crystal structures 497:518 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 0 24 theme activity 37:44 arg1 Modulation 0:9 Modulation 0:9 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 0:127 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 2 25 theme TG 484:485 arg1 activity 487:494 TG activity 484:494 TG activity 484:494 The removal from subsite +3 through mutation of Trp168 to alanine suppressed TG activity, while introduction into subsite +1 through mutation of Gly77 to tryptophan (CrChiA-G77W) enhanced TG activity. 26216755 3 26 from subsite 836:842 arg1 GlcNAc 822:827 GlcNAc 822:827 GlcNAc at the subsite 822:842 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 3 26 from subsite 836:842 arg1 absence 811:817 the absence 807:817 the absence of GlcNAc at the subsite 807:842 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 0 27 theme family 55:60 arg1 chitinase 67:75 plant family GH18 chitinase 49:75 plant family GH18 chitinase 49:75 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 3 28 theme inactive 526:533 arg1 mutant 542:547 an inactive double mutant 523:547 an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) 523:577 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 1 29 from cycad 198:202 arg1 chitinase 179:187 a family GH18 chitinase 165:187 a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 165:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 1 29 from cycad 198:202 arg1 activity 153:160 Transglycosylation (TG) activity 129:160 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 129:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 0 30 theme plant 49:53 arg1 chitinase 67:75 plant family GH18 chitinase 49:75 plant family GH18 chitinase 49:75 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 1 31 theme tryptophan 273:282 arg1 chain 289:293 tryptophan side chain 273:293 a tryptophan side chain 271:293 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 3 32 theme double 535:540 arg1 mutant 542:547 an inactive double mutant 523:547 an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) 523:577 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 4 33 theme aglycon-binding 870:884 arg1 site 886:889 the aglycon-binding site 866:889 the aglycon-binding site 866:889 Aromatic residues in the aglycon-binding site are key determinants of TG activity of GH18 chitinases. 26216755 1 34 theme side 284:287 arg1 chain 289:293 tryptophan side chain 273:293 a tryptophan side chain 271:293 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 3 35 from absence 811:817 arg1 subsite 836:842 the subsite 832:842 the subsite 832:842 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 3 36 theme CrChiA-G77W/E119Q 560:576 arg1 mutant 542:547 an inactive double mutant 523:547 an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) 523:577 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 3 37 theme N-acetylglucosamine 595:613 arg1 residues 615:622 one or two N-acetylglucosamine residues 584:622 one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively 584:667 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 1 38 theme Transglycosylation 129:146 arg1 activity 153:160 Transglycosylation (TG) activity 129:160 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 129:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 1 39 theme Cycas 205:209 arg1 revoluta 211:218 Cycas revoluta 205:218 Cycas revoluta 205:218 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 1 39 theme Cycas 205:209 arg1 cycad 198:202 the cycad 194:202 the cycad 194:202 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 26216755 0 40 theme chitinase 67:75 arg1 activity 37:44 the transglycosylation activity 14:44 the transglycosylation activity of plant family GH18 chitinase 14:75 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 3 41 dep it 754:755 arg1 face-to-face 757:768 face-to-face 757:768 it face-to-face 754:768 The crystal structures of an inactive double mutant of CrChiA (CrChiA-G77W/E119Q) with one or two N-acetylglucosamine residues occupying subsites +1 or +1/+2, respectively, revealed that the Trp77 side chain was oriented toward +1 GlcNAc to be stacked with it face-to-face, but rotated away from subsite +1 in the absence of GlcNAc at the subsite. 26216755 0 42 theme GH18 62:65 arg1 chitinase 67:75 plant family GH18 chitinase 49:75 plant family GH18 chitinase 49:75 Modulation of the transglycosylation activity of plant family GH18 chitinase by removing or introducing a tryptophan side chain. 26216755 1 43 theme TG 149:150 arg1 activity 153:160 Transglycosylation (TG) activity 129:160 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) 129:228 Transglycosylation (TG) activity of a family GH18 chitinase from the cycad, Cycas revoluta, (CrChiA) was modulated by removing or introducing a tryptophan side chain. 24637934 9 0 theme glycoform 1429:1437 arg1 prediction 1411:1420 the in silico prediction 1397:1420 the in silico prediction of the glycoform of a biotherapeutic 1397:1457 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 1 1 theme largest 142:148 arg1 group 150:154 the largest group 138:154 the largest group of the growing number of biologically-derived medicines 138:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 1 1 theme largest 142:148 arg1 number 171:176 the growing number 159:176 the growing number of biologically-derived medicines 159:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 5 2 link N-linked 979:986 arg1 structures 995:1004 N-linked glycan structures 979:1004 N-linked glycan structures of the antibody constant region 979:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 2 3 theme large 289:293 arg1 impact 295:300 a large impact 287:300 a large impact on pharmacokinetics 287:320 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 1 4 theme biologically-derived 181:200 arg1 medicines 202:210 biologically-derived medicines 181:210 biologically-derived medicines 181:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 5 5 theme intracellular 878:890 arg1 concentration 892:904 dynamic intracellular concentration 870:904 their simulated dynamic intracellular concentration profiles 854:913 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 6 theme Golgi 834:838 arg1 apparatus 840:848 the Golgi apparatus 830:848 the Golgi apparatus 830:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 4 7 theme network 652:658 arg1 focus 543:547 The main focus 534:547 The main focus of this work 534:560 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 7 theme network 652:658 arg1 reconstruction 591:604 the mechanistic in silico reconstruction 565:604 the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved 565:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 5 8 from apparatus 840:848 arg1 co-substrates 784:796 the co-substrates 780:796 the co-substrates of the glycosylation process in the Golgi apparatus 780:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 8 from apparatus 840:848 arg1 NSDs 771:774 NSDs 771:774 NSDs 771:774 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 3 9 theme modelling 325:333 arg1 framework 335:343 A modelling framework 323:343 A modelling framework 323:343 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 4 10 theme saturation 705:714 arg1 kinetics 716:723 the saturation kinetics 701:723 the saturation kinetics 701:723 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 9 11 theme bioprocess 1507:1516 arg1 conditions 1518:1527 bioprocess conditions 1507:1527 bioprocess conditions 1507:1527 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 2 12 from impact 295:300 arg1 pharmacokinetics 305:320 pharmacokinetics 305:320 pharmacokinetics 305:320 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 5 13 theme existing 933:940 arg1 model 942:946 an existing model 930:946 an existing model describing the distribution of N-linked glycan structures of the antibody constant region 930:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 14 from co-substrates 784:796 arg1 apparatus 840:848 the Golgi apparatus 830:848 the Golgi apparatus 830:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 1 15 theme number 171:176 arg1 group 150:154 the largest group 138:154 the largest group of the growing number of biologically-derived medicines 138:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 1 15 theme number 171:176 arg1 number 171:176 the growing number 159:176 the growing number of biologically-derived medicines 159:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 0 16 theme antibody 90:97 arg1 glycosylation 99:111 antibody glycosylation 90:111 antibody glycosylation 90:111 Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. 24637934 5 17 from process 819:825 arg1 apparatus 840:848 the Golgi apparatus 830:848 the Golgi apparatus 830:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 18 theme glycan 988:993 arg1 structures 995:1004 N-linked glycan structures 979:1004 N-linked glycan structures of the antibody constant region 979:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 7 19 theme murine 1216:1221 arg1 line 1238:1241 a murine hybridoma cell line 1214:1241 a murine hybridoma cell line 1214:1241 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 6 20 theme modelling 1043:1051 arg1 framework 1053:1061 The modelling framework 1039:1061 The modelling framework 1039:1061 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 5 21 theme glycosylation 805:817 arg1 process 819:825 the glycosylation process 801:825 the glycosylation process in the Golgi apparatus 801:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 3 22 theme extracellular 386:398 arg1 environment 400:410 the extracellular environment 382:410 the extracellular environment 382:410 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 4 23 theme metabolic 741:749 arg1 reactions 751:759 the 60 metabolic reactions 734:759 the 60 metabolic reactions involved 734:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 7 24 theme experimental 1191:1202 arg1 data 1204:1207 experimental data 1191:1207 experimental data from a murine hybridoma cell line 1191:1241 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 3 25 theme intracellular 430:442 arg1 metabolites 444:454 intracellular metabolites 430:454 intracellular metabolites 430:454 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 4 26 theme mechanistic 569:579 arg1 focus 543:547 The main focus 534:547 The main focus of this work 534:560 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 26 theme mechanistic 569:579 arg1 reconstruction 591:604 the mechanistic in silico reconstruction 565:604 the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved 565:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 3 27 theme glycans 479:485 arg1 distribution 463:474 the distribution 459:474 the distribution of glycans on the constant region of an antibody product 459:531 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 9 28 theme first 1378:1382 arg1 step 1384:1387 a first step 1376:1387 a first step towards the in silico prediction of the glycoform of a biotherapeutic 1376:1457 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 0 29 attach linking 53:59 arg2 framework 43:51 a model framework 35:51 a model framework 35:51 Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. 24637934 0 29 attach linking 53:59 arg1 glycosylation 99:111 antibody glycosylation 90:111 antibody glycosylation 90:111 Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. 24637934 0 30 theme model 37:41 arg1 framework 43:51 a model framework 35:51 a model framework 35:51 Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. 24637934 3 31 from effect 420:425 arg1 metabolites 444:454 intracellular metabolites 430:454 intracellular metabolites 430:454 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 2 32 theme glycan 228:233 arg1 structures 235:244 The associated glycan structures 213:244 The associated glycan structures 213:244 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 9 33 gly glycoform 1429:1437 arg1 biotherapeutic 1444:1457 a biotherapeutic 1442:1457 a biotherapeutic 1442:1457 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 9 34 theme in 1401:1402 arg1 prediction 1411:1420 the in silico prediction 1397:1420 the in silico prediction of the glycoform of a biotherapeutic 1397:1457 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 5 35 theme antibody 1013:1020 arg1 region 1031:1036 the antibody constant region 1009:1036 the antibody constant region 1009:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 6 36 theme modified 1133:1140 arg1 kinetics 1148:1155 modified Monod kinetics 1133:1155 modified Monod kinetics 1133:1155 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 4 37 theme nucleotide 613:622 arg1 network 652:658 the nucleotide sugar donor (NSD) metabolic network 609:658 the nucleotide sugar donor (NSD) metabolic network 609:658 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 5 38 theme simulated 860:868 arg1 profiles 906:913 their simulated dynamic intracellular concentration profiles 854:913 their simulated dynamic intracellular concentration profiles 854:913 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 39 theme region 1031:1036 arg1 structures 995:1004 N-linked glycan structures 979:1004 N-linked glycan structures of the antibody constant region 979:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 4 40 theme donor 630:634 arg1 network 652:658 the nucleotide sugar donor (NSD) metabolic network 609:658 the nucleotide sugar donor (NSD) metabolic network 609:658 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 3 41 theme product 525:531 arg1 region 503:508 the constant region 490:508 the constant region of an antibody product 490:531 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 6 42 theme population 1110:1119 arg1 dynamics 1089:1096 the growth dynamics 1078:1096 the growth dynamics of the cell population 1078:1119 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 2 43 contain have 282:285 arg1 distribution 256:267 their distribution 250:267 their distribution 250:267 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 2 43 contain have 282:285 arg2 impact 295:300 a large impact 287:300 a large impact on pharmacokinetics 287:320 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 2 43 contain have 282:285 arg1 structures 235:244 The associated glycan structures 213:244 The associated glycan structures 213:244 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 9 44 theme product 1545:1551 arg1 quality 1553:1559 product quality 1545:1559 product quality 1545:1559 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 5 45 theme dynamic 870:876 arg1 concentration 892:904 dynamic intracellular concentration 870:904 their simulated dynamic intracellular concentration profiles 854:913 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 9 46 theme biotherapeutic 1444:1457 arg1 glycoform 1429:1437 the glycoform 1425:1437 the glycoform of a biotherapeutic 1425:1457 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 5 47 attach linked 920:925 arg2 profiles 906:913 their simulated dynamic intracellular concentration profiles 854:913 their simulated dynamic intracellular concentration profiles 854:913 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 47 attach linked 920:925 arg1 model 942:946 an existing model 930:946 an existing model describing the distribution of N-linked glycan structures of the antibody constant region 930:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 1 48 theme medicines 202:210 arg1 number 171:176 the growing number 159:176 the growing number of biologically-derived medicines 159:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 8 49 theme modelling 1260:1268 arg1 platform 1270:1277 a modelling platform 1258:1277 a modelling platform which is able to describe the product glycoform based on extracellular conditions 1258:1359 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 8 49 theme modelling 1260:1268 arg1 able 1288:1291 able 1288:1291 able 1288:1291 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 8 49 theme modelling 1260:1268 arg1 result 1248:1253 The result 1244:1253 The result 1244:1253 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 4 50 theme NSD 637:639 arg1 network 652:658 the nucleotide sugar donor (NSD) metabolic network 609:658 the nucleotide sugar donor (NSD) metabolic network 609:658 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 3 51 from link 372:375 arg1 metabolites 444:454 intracellular metabolites 430:454 intracellular metabolites 430:454 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 9 52 dep in 1401:1402 arg1 silico 1404:1409 silico 1404:1409 silico 1404:1409 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 4 53 theme metabolic 642:650 arg1 network 652:658 the nucleotide sugar donor (NSD) metabolic network 609:658 the nucleotide sugar donor (NSD) metabolic network 609:658 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 7 54 from line 1238:1241 arg1 data 1204:1207 experimental data 1191:1207 experimental data from a murine hybridoma cell line 1191:1241 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 9 55 theme conditions 1518:1527 arg1 optimisation 1491:1502 the optimisation 1487:1502 the optimisation of bioprocess conditions 1487:1527 It represents a first step towards the in silico prediction of the glycoform of a biotherapeutic and provides a platform for the optimisation of bioprocess conditions with respect to product quality. 24637934 8 56 theme product 1309:1315 arg1 glycoform 1317:1325 the product glycoform 1305:1325 the product glycoform based on extracellular conditions 1305:1359 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 0 57 theme extracellular 61:73 arg1 metabolites 75:85 extracellular metabolites 61:85 extracellular metabolites 61:85 Towards controlling the glycoform: a model framework linking extracellular metabolites to antibody glycosylation. 24637934 5 58 theme concentration 892:904 arg1 profiles 906:913 their simulated dynamic intracellular concentration profiles 854:913 their simulated dynamic intracellular concentration profiles 854:913 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 4 59 theme species 675:681 arg1 mass 683:686 34 species mass balances and the saturation kinetics rates 672:729 mass 683:686 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 7 60 theme cell 1233:1236 arg1 line 1238:1241 a murine hybridoma cell line 1214:1241 a murine hybridoma cell line 1214:1241 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 1 61 theme growing 163:169 arg1 number 171:176 the growing number 159:176 the growing number of biologically-derived medicines 159:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 4 62 dep in 581:582 arg1 silico 584:589 silico 584:589 silico 584:589 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 63 theme main 538:541 arg1 focus 543:547 The main focus 534:547 The main focus of this work 534:560 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 63 theme main 538:541 arg1 reconstruction 591:604 the mechanistic in silico reconstruction 565:604 the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved 565:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 8 64 theme extracellular 1336:1348 arg1 conditions 1350:1359 extracellular conditions 1336:1359 extracellular conditions 1336:1359 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 7 65 theme hybridoma 1223:1231 arg1 line 1238:1241 a murine hybridoma cell line 1214:1241 a murine hybridoma cell line 1214:1241 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 8 66 gly glycoform 1317:1325 arg1 product 1309:1315 the product glycoform 1305:1325 the product glycoform based on extracellular conditions 1305:1359 The result is a modelling platform which is able to describe the product glycoform based on extracellular conditions. 24637934 5 67 theme N-linked 979:986 arg1 structures 995:1004 N-linked glycan structures 979:1004 N-linked glycan structures of the antibody constant region 979:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 4 68 theme reactions 751:759 arg1 kinetics 716:723 the saturation kinetics 701:723 the saturation kinetics 701:723 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 68 theme reactions 751:759 arg1 mass 683:686 34 species mass balances and the saturation kinetics rates 672:729 mass 683:686 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 3 69 from environment 400:410 arg1 link 372:375 a link 370:375 a link from the extracellular environment 370:410 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 3 69 from environment 400:410 arg1 effect 420:425 its effect 416:425 its effect on intracellular metabolites 416:454 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 4 70 dep mass 683:686 arg1 balances 688:695 balances 688:695 balances 688:695 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 70 dep mass 683:686 arg1 rates 725:729 rates 725:729 rates 725:729 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 6 71 theme cell 1105:1108 arg1 population 1110:1119 the cell population 1101:1119 the cell population 1101:1119 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 1 72 link biologically-derived 181:200 arg1 medicines 202:210 biologically-derived medicines 181:210 biologically-derived medicines 181:210 Glycoproteins represent the largest group of the growing number of biologically-derived medicines. 24637934 5 73 theme process 819:825 arg1 co-substrates 784:796 the co-substrates 780:796 the co-substrates of the glycosylation process in the Golgi apparatus 780:848 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 5 73 theme process 819:825 arg1 NSDs 771:774 NSDs 771:774 NSDs 771:774 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 6 74 theme growth 1082:1087 arg1 dynamics 1089:1096 the growth dynamics 1078:1096 the growth dynamics of the cell population 1078:1119 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 4 75 theme in 581:582 arg1 focus 543:547 The main focus 534:547 The main focus of this work 534:560 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 75 theme in 581:582 arg1 reconstruction 591:604 the mechanistic in silico reconstruction 565:604 the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved 565:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 5 76 theme structures 995:1004 arg1 distribution 963:974 the distribution 959:974 the distribution of N-linked glycan structures of the antibody constant region 959:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 3 77 theme constant 494:501 arg1 region 503:508 the constant region 490:508 the constant region of an antibody product 490:531 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 7 78 theme Simulation 1158:1167 arg1 results 1169:1175 Simulation results 1158:1175 Simulation results 1158:1175 Simulation results match well to experimental data from a murine hybridoma cell line. 24637934 4 79 theme work 557:560 arg1 focus 543:547 The main focus 534:547 The main focus of this work 534:560 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 4 79 theme work 557:560 arg1 reconstruction 591:604 the mechanistic in silico reconstruction 565:604 the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved 565:768 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 3 80 theme antibody 516:523 arg1 product 525:531 an antibody product 513:531 an antibody product 513:531 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 24637934 6 81 theme Monod 1142:1146 arg1 kinetics 1148:1155 modified Monod kinetics 1133:1155 modified Monod kinetics 1133:1155 The modelling framework also describes the growth dynamics of the cell population by means of modified Monod kinetics. 24637934 4 82 theme sugar 624:628 arg1 network 652:658 the nucleotide sugar donor (NSD) metabolic network 609:658 the nucleotide sugar donor (NSD) metabolic network 609:658 The main focus of this work is the mechanistic in silico reconstruction of the nucleotide sugar donor (NSD) metabolic network by means of 34 species mass balances and the saturation kinetics rates of the 60 metabolic reactions involved. 24637934 2 83 theme associated 217:226 arg1 structures 235:244 The associated glycan structures 213:244 The associated glycan structures 213:244 The associated glycan structures and their distribution are known to have a large impact on pharmacokinetics. 24637934 5 84 theme constant 1022:1029 arg1 region 1031:1036 the antibody constant region 1009:1036 the antibody constant region 1009:1036 NSDs are the co-substrates of the glycosylation process in the Golgi apparatus and their simulated dynamic intracellular concentration profiles were linked to an existing model describing the distribution of N-linked glycan structures of the antibody constant region. 24637934 3 85 from distribution 463:474 arg1 region 503:508 the constant region 490:508 the constant region of an antibody product 490:531 A modelling framework was developed to provide a link from the extracellular environment and its effect on intracellular metabolites to the distribution of glycans on the constant region of an antibody product. 25002537 3 0 from genes 1044:1048 arg1 locus 1076:1080 each corresponding cps locus 1053:1080 each corresponding cps locus 1053:1080 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 1 theme CPS39 1347:1351 arg1 synthesis 1334:1342 the synthesis 1330:1342 the synthesis of CPS39 1330:1351 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 4 2 theme only 1087:1090 arg1 exception 1092:1100 The only exception 1083:1100 The only exception 1083:1100 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 5 3 theme α1-1 1536:1539 arg1 transfer 1541:1548 α1-1 transfer 1536:1548 α1-1 transfer of Galp to ribitol-5-phosphate 1536:1579 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 5 4 dep wefM 1482:1485 arg1 i.e. 1476:1479 i.e. 1476:1479 i.e. 1476:1479 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 3 5 theme specific 1035:1042 arg1 genes 1044:1048 specific genes 1035:1048 specific genes in each corresponding cps locus 1035:1080 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 6 theme composition 728:738 arg1 analyses 740:747 glycosyl composition analyses 719:747 glycosyl composition analyses 719:747 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 7 theme repeat 773:778 arg1 structures 785:794 the following repeat unit structures 759:794 the following repeat unit structures 759:794 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 8 theme High-resolution 575:589 arg1 NMR 633:635 NMR 633:635 NMR 633:635 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 8 theme High-resolution 575:589 arg1 resonance 622:630 High-resolution heteronuclear nuclear magnetic resonance 575:630 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 9 theme corresponding 1058:1070 arg1 locus 1076:1080 each corresponding cps locus 1053:1080 each corresponding cps locus 1053:1080 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 0 10 theme CPS34 101:105 arg1 structures 9:18 Chemical structures 0:18 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 0:105 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 1 11 theme Structural 194:203 arg1 characterization 205:220 Structural characterization 194:220 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) 194:279 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 1 11 theme Structural 194:203 arg1 prerequisite 286:297 a prerequisite 284:297 a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes 284:392 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 3 12 theme resonance 622:630 arg1 spectroscopy 638:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 5 13 dep resembled 1436:1444 arg1 related 1406:1412 related 1406:1412 related to wcrC 1406:1420 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 0 14 theme magnetic 132:139 arg1 resonance 141:149 nuclear magnetic resonance 124:149 nuclear magnetic resonance spectroscopy 124:162 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 15 theme nuclear 605:611 arg1 NMR 633:635 NMR 633:635 NMR 633:635 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 15 theme nuclear 605:611 arg1 resonance 622:630 High-resolution heteronuclear nuclear magnetic resonance 575:630 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 16 theme serotypes 809:817 arg1 structures 785:794 the following repeat unit structures 759:794 the following repeat unit structures 759:794 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 6 17 from residue 1681:1687 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 17 from residue 1681:1687 arg1 sites 1736:1740 important sites 1726:1740 important sites of CPS structural and genetic diversity 1726:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 17 from residue 1681:1687 arg1 structural 1749:1758 structural 1749:1758 structural 1749:1758 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 17 from residue 1681:1687 arg1 linkages 1623:1630 linkages 1623:1630 linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) 1623:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 3 18 theme Common 888:893 arg1 features 917:924 [structure: see text] Common and unique structural features 866:924 [structure: see text] Common and unique structural features of these polysaccharides 866:949 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 18 theme Common 888:893 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 19 theme Gal 1183:1185 arg1 Galp 1199:1202 Galp 1199:1202 Galp 1199:1202 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 4 19 theme Gal 1183:1185 arg1 pyranoside 1187:1196 Gal pyranoside 1183:1196 Gal pyranoside (Galp) 1183:1203 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 3 20 theme published 665:673 arg1 structure 675:683 the published structure 661:683 the published structure of CPS34 661:692 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 1 21 theme different 374:382 arg1 serotypes 384:392 different serotypes 374:392 different serotypes 374:392 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 2 22 theme related 527:533 arg1 CPS34 549:553 CPS34 549:553 CPS34 549:553 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 6 23 theme recent 1598:1603 arg1 findings 1605:1612 other recent findings 1592:1612 other recent findings 1592:1612 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 3 24 theme unique 899:904 arg1 features 917:924 [structure: see text] Common and unique structural features 866:924 [structure: see text] Common and unique structural features of these polysaccharides 866:949 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 24 theme unique 899:904 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 5 25 from gene 1372:1375 arg1 locus 1390:1394 the cps39 locus 1380:1394 the cps39 locus 1380:1394 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 2 26 theme serogroup 469:477 arg1 studies 440:446 comparative structural studies 417:446 comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) 417:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 6 27 theme adjacent 1692:1699 arg1 furanoside 1705:1714 adjacent Gal furanoside 1692:1714 adjacent Gal furanoside (Galf) 1692:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 27 theme adjacent 1692:1699 arg1 Galf 1717:1720 Galf 1717:1720 Galf 1717:1720 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 2 28 theme current 402:408 arg1 study 410:414 the current study 398:414 the current study 398:414 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 0 29 theme Chemical 0:7 arg1 structures 9:18 Chemical structures 0:18 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 0:105 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 2 30 theme pneumoniae 454:463 arg1 serogroup 469:477 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 6 31 theme diversity 1772:1780 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 31 theme diversity 1772:1780 arg1 sites 1736:1740 important sites 1726:1740 important sites of CPS structural and genetic diversity 1726:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 31 theme diversity 1772:1780 arg1 structural 1749:1758 structural 1749:1758 structural 1749:1758 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 31 theme diversity 1772:1780 arg1 linkages 1623:1630 linkages 1623:1630 linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) 1623:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 5 32 theme oralis 1505:1510 arg1 wefM 1482:1485 wefM 1482:1485 wefM 1482:1485 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 5 32 theme oralis 1505:1510 arg1 gene 1470:1473 a previously identified gene 1446:1473 a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate 1446:1579 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 4 33 from ribitol-5-phosphate 1208:1226 arg1 synthesis 1235:1243 the synthesis 1231:1243 the synthesis of CPS10A, CPS47F, and CPS34 1231:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 2 34 theme structural 429:438 arg1 studies 440:446 comparative structural studies 417:446 comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) 417:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 2 35 dep S. 535:536 arg1 pneumoniae 538:547 pneumoniae 538:547 pneumoniae 538:547 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 0 36 theme polysaccharide 57:70 arg1 CPS39 81:85 CPS39 81:85 CPS39 81:85 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 0 36 theme polysaccharide 57:70 arg1 type 72:75 Streptococcus pneumoniae capsular polysaccharide type 39 23:78 Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39) 23:86 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 37 theme different 962:970 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 1 38 theme Streptococcus 225:237 arg1 CPS 276:278 CPS 276:278 CPS 276:278 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 1 38 theme Streptococcus 225:237 arg1 polysaccharides 259:273 Streptococcus pneumoniae capsular polysaccharides 225:273 Streptococcus pneumoniae capsular polysaccharides (CPS) 225:279 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 5 39 theme Galp 1553:1556 arg1 transfer 1541:1548 α1-1 transfer 1536:1548 α1-1 transfer of Galp to ribitol-5-phosphate 1536:1579 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 4 40 theme CPS34 1268:1272 arg1 synthesis 1235:1243 the synthesis 1231:1243 the synthesis of CPS10A, CPS47F, and CPS34 1231:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 1 41 theme capsular 250:257 arg1 CPS 276:278 CPS 276:278 CPS 276:278 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 1 41 theme capsular 250:257 arg1 polysaccharides 259:273 Streptococcus pneumoniae capsular polysaccharides 225:273 Streptococcus pneumoniae capsular polysaccharides (CPS) 225:279 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 3 42 theme glycosyl 719:726 arg1 analyses 740:747 glycosyl composition analyses 719:747 glycosyl composition analyses 719:747 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 43 with conjunction 702:712 arg1 analyses 740:747 glycosyl composition analyses 719:747 glycosyl composition analyses 719:747 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 44 theme Gal 1300:1302 arg1 transfer 1288:1295 α1-1 transfer 1283:1295 α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39 1283:1351 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 3 45 theme cps 1072:1074 arg1 locus 1076:1080 each corresponding cps locus 1053:1080 each corresponding cps locus 1053:1080 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 46 theme α1-2 1166:1169 arg1 transfer 1171:1178 the α1-2 transfer 1162:1178 the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 1162:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 3 47 theme unit 780:783 arg1 structures 785:794 the following repeat unit structures 759:794 the following repeat unit structures 759:794 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 48 theme following 763:771 arg1 structures 785:794 the following repeat unit structures 759:794 the following repeat unit structures 759:794 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 0 49 theme nuclear 124:130 arg1 resonance 141:149 nuclear magnetic resonance 124:149 nuclear magnetic resonance spectroscopy 124:162 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 50 theme magnetic 613:620 arg1 NMR 633:635 NMR 633:635 NMR 633:635 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 50 theme magnetic 613:620 arg1 resonance 622:630 High-resolution heteronuclear nuclear magnetic resonance 575:630 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 1 51 dep Streptococcus 225:237 arg1 pneumoniae 239:248 pneumoniae 239:248 pneumoniae 239:248 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 3 52 theme other 803:807 arg1 serotypes 809:817 the other serotypes 799:817 the other serotypes 799:817 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 0 53 theme resonance 141:149 arg1 spectroscopy 151:162 nuclear magnetic resonance spectroscopy 124:162 nuclear magnetic resonance spectroscopy 124:162 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 54 theme heteronuclear 591:603 arg1 NMR 633:635 NMR 633:635 NMR 633:635 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 54 theme heteronuclear 591:603 arg1 resonance 622:630 High-resolution heteronuclear nuclear magnetic resonance 575:630 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy 575:649 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 6 55 theme important 1726:1734 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 55 theme important 1726:1734 arg1 sites 1736:1740 important sites 1726:1740 important sites of CPS structural and genetic diversity 1726:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 55 theme important 1726:1734 arg1 structural 1749:1758 structural 1749:1758 structural 1749:1758 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 55 theme important 1726:1734 arg1 linkages 1623:1630 linkages 1623:1630 linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) 1623:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 4 56 theme pyranoside 1187:1196 arg1 transfer 1171:1178 the α1-2 transfer 1162:1178 the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 1162:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 3 57 dep confirmed 651:659 arg1 revealed 750:757 revealed 750:757 revealed the following repeat unit structures of the other serotypes, which have not been previously characterized 750:863 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 58 theme CPS10A 1248:1253 arg1 synthesis 1235:1243 the synthesis 1231:1243 the synthesis of CPS10A, CPS47F, and CPS34 1231:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 0 59 theme CPS47F 89:94 arg1 structures 9:18 Chemical structures 0:18 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 0:105 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 60 dep [structure 866:875 arg1 see 878:880 see 878:880 see text 878:885 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 4 61 theme CPS47F 1256:1261 arg1 synthesis 1235:1243 the synthesis 1231:1243 the synthesis of CPS10A, CPS47F, and CPS34 1231:1272 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 6 62 theme Gal 1701:1703 arg1 furanoside 1705:1714 adjacent Gal furanoside 1692:1714 adjacent Gal furanoside (Galf) 1692:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 62 theme Gal 1701:1703 arg1 Galf 1717:1720 Galf 1717:1720 Galf 1717:1720 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 4 63 from ribitol-5-phosphate 1307:1325 arg1 synthesis 1334:1342 the synthesis 1330:1342 the synthesis of CPS39 1330:1351 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 6 64 theme other 1592:1596 arg1 findings 1605:1612 other recent findings 1592:1612 other recent findings 1592:1612 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 3 65 theme structural 906:915 arg1 features 917:924 [structure: see text] Common and unique structural features 866:924 [structure: see text] Common and unique structural features of these polysaccharides 866:949 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 65 theme structural 906:915 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 2 66 theme CPS 465:467 arg1 serogroup 469:477 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 5 67 theme identified 1459:1468 arg1 wefM 1482:1485 wefM 1482:1485 wefM 1482:1485 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 5 67 theme identified 1459:1468 arg1 gene 1470:1473 a previously identified gene 1446:1473 a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate 1446:1579 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 0 68 theme Streptococcus 23:35 arg1 polysaccharide 57:70 Streptococcus pneumoniae capsular polysaccharide 23:70 Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39) 23:86 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 2 69 dep related 527:533 arg1 S. 535:536 S. 535:536 S. 535:536 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 2 70 theme S. 451:452 arg1 serogroup 469:477 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 S. pneumoniae CPS serogroup 10 (CPS10) 451:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 6 71 theme genetic 1764:1770 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 4 72 theme α1-1 1283:1286 arg1 transfer 1288:1295 α1-1 transfer 1283:1295 α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39 1283:1351 The only exception involved the gene designated wcrC, which is associated with the α1-2 transfer of Gal pyranoside (Galp) to ribitol-5-phosphate in the synthesis of CPS10A, CPS47F, and CPS34 but with α1-1 transfer of Gal to ribitol-5-phosphate in the synthesis of CPS39. 25002537 0 73 theme capsular 48:55 arg1 polysaccharide 57:70 Streptococcus pneumoniae capsular polysaccharide 23:70 Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39) 23:86 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 3 74 theme CPS34 688:692 arg1 structure 675:683 the published structure 661:683 the published structure of CPS34 661:692 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 75 theme polysaccharides 935:949 arg1 features 917:924 [structure: see text] Common and unique structural features 866:924 [structure: see text] Common and unique structural features of these polysaccharides 866:949 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 3 75 theme polysaccharides 935:949 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 6 76 from α-Galp 1637:1642 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 76 from α-Galp 1637:1642 arg1 sites 1736:1740 important sites 1726:1740 important sites of CPS structural and genetic diversity 1726:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 76 from α-Galp 1637:1642 arg1 structural 1749:1758 structural 1749:1758 structural 1749:1758 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 76 from α-Galp 1637:1642 arg1 linkages 1623:1630 linkages 1623:1630 linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) 1623:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 77 theme structural 1749:1758 arg1 diversity 1772:1780 CPS structural and genetic diversity 1745:1780 diversity 1772:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 77 theme structural 1749:1758 arg1 sites 1736:1740 important sites 1726:1740 important sites of CPS structural and genetic diversity 1726:1780 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 77 theme structural 1749:1758 arg1 structural 1749:1758 structural 1749:1758 structural 1749:1758 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 6 77 theme structural 1749:1758 arg1 linkages 1623:1630 linkages 1623:1630 linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) 1623:1721 These and other recent findings identify linkages from α-Galp to ribitol-5-phosphate and from this residue to adjacent Gal furanoside (Galf) as important sites of CPS structural and genetic diversity. 25002537 2 78 theme comparative 417:427 arg1 studies 440:446 comparative structural studies 417:446 comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) 417:488 In the current study, comparative structural studies of S. pneumoniae CPS serogroup 10 (CPS10) were extended to include genetically related S. pneumoniae CPS34, CPS39, and CPS47F. 25002537 0 79 theme type 72:75 arg1 structures 9:18 Chemical structures 0:18 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 0:105 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 1 80 theme antigenic 319:327 arg1 relationships 341:353 both antigenic and genetic relationships 314:353 both antigenic and genetic relationships that exist between different serotypes 314:392 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 3 81 theme O-acetylation 985:997 arg1 positions 972:980 different positions 962:980 different positions of O-acetylation 962:997 High-resolution heteronuclear nuclear magnetic resonance (NMR) spectroscopy confirmed the published structure of CPS34 and, in conjunction with glycosyl composition analyses, revealed the following repeat unit structures of the other serotypes, which have not been previously characterized: [structure: see text] Common and unique structural features of these polysaccharides, including different positions of O-acetylation, were unambiguously associated with specific genes in each corresponding cps locus. 25002537 5 82 theme cps39 1384:1388 arg1 locus 1390:1394 the cps39 locus 1380:1394 the cps39 locus 1380:1394 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 1 83 theme genetic 333:339 arg1 relationships 341:353 both antigenic and genetic relationships 314:353 both antigenic and genetic relationships that exist between different serotypes 314:392 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 0 84 dep Streptococcus 23:35 arg1 pneumoniae 37:46 pneumoniae 37:46 pneumoniae 37:46 Chemical structures of Streptococcus pneumoniae capsular polysaccharide type 39 (CPS39), CPS47F, and CPS34 characterized by nuclear magnetic resonance spectroscopy and their relation to CPS10A. 25002537 5 85 theme Streptococcus 1491:1503 arg1 oralis 1505:1510 Streptococcus oralis 1491:1510 Streptococcus oralis 1491:1510 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 5 86 theme corresponding 1358:1370 arg1 gene 1372:1375 The corresponding gene 1354:1375 The corresponding gene in the cps39 locus 1354:1394 The corresponding gene in the cps39 locus, although related to wcrC, more closely resembled a previously identified gene (i.e., wefM) of Streptococcus oralis that is associated with α1-1 transfer of Galp to ribitol-5-phosphate. 25002537 1 87 theme polysaccharides 259:273 arg1 characterization 205:220 Structural characterization 194:220 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) 194:279 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 25002537 1 87 theme polysaccharides 259:273 arg1 prerequisite 286:297 a prerequisite 284:297 a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes 284:392 Structural characterization of Streptococcus pneumoniae capsular polysaccharides (CPS) is a prerequisite for unraveling both antigenic and genetic relationships that exist between different serotypes. 26926155 4 0 theme efficient 575:583 arg1 strategy 585:592 a highly efficient strategy 566:592 a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk 566:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 5 1 theme remaining 1029:1037 arg1 groups 1047:1052 the remaining hydroxy groups 1025:1052 the remaining hydroxy groups with acetyl groups 1025:1071 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 4 2 theme 10-step 685:691 arg1 conversions 702:712 only 9- or 10-step chemical conversions 674:712 only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk 674:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 7 3 with acceptors 1219:1227 arg1 thioglycoside 1259:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 7 3 with acceptors 1219:1227 arg1 steps 1301:1305 subsequent deprotection steps 1277:1305 subsequent deprotection steps 1277:1305 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 4 4 theme representative 608:621 arg1 types 627:631 the representative two types 604:631 the representative two types of triantennary oligosaccharides 604:664 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 7 5 theme protected 1234:1242 arg1 thioglycoside 1259:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 4 6 theme egg 811:813 arg1 yolk 815:818 egg yolk 811:818 egg yolk 811:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 0 7 theme Natural 118:124 arg1 Source 126:131 a Natural Source 116:131 a Natural Source 116:131 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 7 8 theme subsequent 1277:1286 arg1 steps 1301:1305 subsequent deprotection steps 1277:1305 subsequent deprotection steps 1277:1305 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 5 9 theme core 916:919 arg1 mannosides 921:930 two core mannosides 912:930 two core mannosides 912:930 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 7 10 theme deprotection 1288:1299 arg1 steps 1301:1305 subsequent deprotection steps 1277:1305 subsequent deprotection steps 1277:1305 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 6 11 theme Selective 1074:1082 arg1 removal 1084:1090 Selective removal 1074:1090 Selective removal of one of the benzylidene acetals 1074:1124 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 4 12 theme chemical 693:700 arg1 conversions 702:712 only 9- or 10-step chemical conversions 674:712 only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk 674:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 3 13 theme proteins 427:434 arg1 syntheses 391:399 syntheses 391:399 syntheses of oligosaccharide-linked proteins 391:434 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 4 14 theme homogeneous 789:799 arg1 form 801:804 exceptionally homogeneous form 775:804 exceptionally homogeneous form 775:804 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 1 15 theme oligosaccharides 198:213 arg1 Attachment 184:193 Attachment 184:193 Attachment of oligosaccharides to proteins 184:225 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 1 15 theme oligosaccharides 198:213 arg1 modification 257:268 a major post-translational modification 230:268 a major post-translational modification 230:268 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 6 16 theme acetals 1118:1124 arg1 acetals 1118:1124 the benzylidene acetals 1102:1124 the benzylidene acetals 1102:1124 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 6 16 theme acetals 1118:1124 arg1 one 1095:1097 one 1095:1097 one 1095:1097 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 5 17 theme hydroxy 1039:1045 arg1 groups 1047:1052 the remaining hydroxy groups 1025:1052 the remaining hydroxy groups with acetyl groups 1025:1071 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 3 18 theme tetra-antennary 533:547 arg1 forms 549:553 tetra-antennary forms 533:553 tetra-antennary forms 533:553 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 7 19 theme oligosaccharides 1367:1382 arg1 types 1332:1336 two types 1328:1336 two types of complex-type triantennary oligosaccharides 1328:1382 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 1 20 attach Attachment 184:193 arg1 proteins 218:225 proteins 218:225 proteins 218:225 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 1 20 attach Attachment 184:193 arg2 oligosaccharides 198:213 oligosaccharides 198:213 oligosaccharides 198:213 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 5 21 with groups 1047:1052 arg1 groups 1066:1071 acetyl groups 1059:1071 acetyl groups 1059:1071 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 6 22 theme benzylidene 1106:1116 arg1 acetals 1118:1124 the benzylidene acetals 1102:1124 the benzylidene acetals 1102:1124 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 3 23 theme diverse 518:524 arg1 di- 526:528 diverse di- 518:528 diverse di- to tetra-antennary forms 518:553 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 3 24 theme inherent 475:482 arg1 structures 496:505 their inherent complicated structures 469:505 their inherent complicated structures 469:505 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 3 24 theme inherent 475:482 arg1 di- 526:528 diverse di- 518:528 diverse di- to tetra-antennary forms 518:553 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 5 25 theme benzylidene 826:836 arg1 acetals 838:844 Four benzylidene acetals 821:844 Four benzylidene acetals 821:844 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 7 26 theme Gal-β-1,4-GlcN 1244:1257 arg1 thioglycoside 1259:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 protected Gal-β-1,4-GlcN thioglycoside 1234:1271 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 0 27 theme Complex-Type 24:35 arg1 Oligosaccharides 50:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 4 28 theme oligosaccharides 649:664 arg1 types 627:631 the representative two types 604:631 the representative two types of triantennary oligosaccharides 604:664 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 0 29 theme Chemical 146:153 arg1 Glycosylation 169:181 Selective Chemical and Enzymatic Glycosylation 136:181 Selective Chemical and Enzymatic Glycosylation 136:181 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 2 30 theme oligosaccharides 364:379 arg1 functions 345:353 the functions 341:353 the functions of these oligosaccharides 341:379 Chemical syntheses of oligosaccharides have contributed to clarifying the functions of these oligosaccharides. 26926155 0 31 theme Intact 17:22 arg1 Oligosaccharides 50:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 4 32 theme 9- 679:680 arg1 conversions 702:712 only 9- or 10-step chemical conversions 674:712 only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk 674:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 0 33 theme Selective 136:144 arg1 Glycosylation 169:181 Selective Chemical and Enzymatic Glycosylation 136:181 Selective Chemical and Enzymatic Glycosylation 136:181 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 4 34 attach isolated 763:770 arg2 oligosaccharide 733:747 a biantennary oligosaccharide 719:747 a biantennary oligosaccharide 719:747 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 4 34 attach isolated 763:770 arg1 yolk 815:818 egg yolk 811:818 egg yolk 811:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 3 35 link oligosaccharide-linked 404:425 arg1 proteins 427:434 oligosaccharide-linked proteins 404:434 oligosaccharide-linked proteins 404:434 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 1 36 theme major 232:236 arg1 Attachment 184:193 Attachment 184:193 Attachment of oligosaccharides to proteins 184:225 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 1 36 theme major 232:236 arg1 modification 257:268 a major post-translational modification 230:268 a major post-translational modification 230:268 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 0 37 theme Oligosaccharides 50:65 arg1 Semisynthesis 0:12 Semisynthesis 0:12 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 0:182 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 6 38 theme one 1095:1097 arg1 removal 1084:1090 Selective removal 1074:1090 Selective removal of one of the benzylidene acetals 1074:1124 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 0 39 theme Enzymatic 159:167 arg1 Glycosylation 169:181 Selective Chemical and Enzymatic Glycosylation 136:181 Selective Chemical and Enzymatic Glycosylation 136:181 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 5 40 theme hydroxy 983:989 arg1 groups 991:996 24 hydroxy groups 980:996 24 hydroxy groups 980:996 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 1 41 theme post-translational 238:255 arg1 Attachment 184:193 Attachment 184:193 Attachment of oligosaccharides to proteins 184:225 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 1 41 theme post-translational 238:255 arg1 modification 257:268 a major post-translational modification 230:268 a major post-translational modification 230:268 Attachment of oligosaccharides to proteins is a major post-translational modification. 26926155 0 42 theme Triantennary 37:48 arg1 Oligosaccharides 50:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Intact Complex-Type Triantennary Oligosaccharides 17:65 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 7 43 theme individual 1208:1217 arg1 acceptors 1219:1227 the individual acceptors 1204:1227 the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps 1204:1305 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 0 44 from Oligosaccharide 86:100 arg1 Semisynthesis 0:12 Semisynthesis 0:12 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 0:182 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 5 45 theme biantennary 939:949 arg1 asialononasaccharide 951:970 the biantennary asialononasaccharide 935:970 the biantennary asialononasaccharide bearing 24 hydroxy groups 935:996 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 6 46 theme acceptors 1172:1180 arg1 types 1135:1139 two types 1131:1139 two types of suitably protected glycosyl acceptors 1131:1180 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 3 47 theme complicated 484:494 arg1 structures 496:505 their inherent complicated structures 469:505 their inherent complicated structures 469:505 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 3 47 theme complicated 484:494 arg1 di- 526:528 diverse di- 518:528 diverse di- to tetra-antennary forms 518:553 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 3 48 theme oligosaccharide-linked 404:425 arg1 proteins 427:434 oligosaccharide-linked proteins 404:434 oligosaccharide-linked proteins 404:434 However, syntheses of oligosaccharide-linked proteins are still challenging because of their inherent complicated structures, including diverse di- to tetra-antennary forms. 26926155 2 49 theme Chemical 271:278 arg1 syntheses 280:288 Chemical syntheses 271:288 Chemical syntheses of oligosaccharides 271:308 Chemical syntheses of oligosaccharides have contributed to clarifying the functions of these oligosaccharides. 26926155 5 50 theme groups 1047:1052 arg1 protection 1011:1020 protection 1011:1020 protection of the remaining hydroxy groups with acetyl groups 1011:1071 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 4 51 theme triantennary 636:647 arg1 oligosaccharides 649:664 triantennary oligosaccharides 636:664 triantennary oligosaccharides 636:664 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 0 52 attach Isolated 102:109 arg1 Source 126:131 a Natural Source 116:131 a Natural Source 116:131 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 0 52 attach Isolated 102:109 arg2 Oligosaccharide 86:100 a Biantennary Oligosaccharide 72:100 a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation 72:181 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 7 53 theme complex-type 1341:1352 arg1 oligosaccharides 1367:1382 complex-type triantennary oligosaccharides 1341:1382 complex-type triantennary oligosaccharides 1341:1382 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 5 54 theme terminal 882:889 arg1 galactosides 895:906 the terminal two galactosides 878:906 the terminal two galactosides 878:906 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 5 55 theme acetyl 1059:1064 arg1 groups 1066:1071 acetyl groups 1059:1071 acetyl groups 1059:1071 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 6 56 theme glycosyl 1163:1170 arg1 acceptors 1172:1180 suitably protected glycosyl acceptors 1144:1180 suitably protected glycosyl acceptors 1144:1180 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 5 57 theme asialononasaccharide 951:970 arg1 mannosides 921:930 two core mannosides 912:930 two core mannosides 912:930 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 5 57 theme asialononasaccharide 951:970 arg1 galactosides 895:906 the terminal two galactosides 878:906 the terminal two galactosides 878:906 Four benzylidene acetals were successfully introduced to the terminal two galactosides and two core mannosides of the biantennary asialononasaccharide bearing 24 hydroxy groups, followed by protection of the remaining hydroxy groups with acetyl groups. 26926155 6 58 theme protected 1153:1161 arg1 acceptors 1172:1180 suitably protected glycosyl acceptors 1144:1180 suitably protected glycosyl acceptors 1144:1180 Selective removal of one of the benzylidene acetals gave two types of suitably protected glycosyl acceptors. 26926155 4 59 from oligosaccharide 733:747 arg1 conversions 702:712 only 9- or 10-step chemical conversions 674:712 only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk 674:818 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 26926155 0 60 theme Biantennary 74:84 arg1 Oligosaccharide 86:100 a Biantennary Oligosaccharide 72:100 a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation 72:181 Semisynthesis of Intact Complex-Type Triantennary Oligosaccharides from a Biantennary Oligosaccharide Isolated from a Natural Source by Selective Chemical and Enzymatic Glycosylation. 26926155 2 61 theme oligosaccharides 293:308 arg1 syntheses 280:288 Chemical syntheses 271:288 Chemical syntheses of oligosaccharides 271:308 Chemical syntheses of oligosaccharides have contributed to clarifying the functions of these oligosaccharides. 26926155 7 62 theme triantennary 1354:1365 arg1 oligosaccharides 1367:1382 complex-type triantennary oligosaccharides 1341:1382 complex-type triantennary oligosaccharides 1341:1382 Glycosylation toward the individual acceptors with protected Gal-β-1,4-GlcN thioglycoside and subsequent deprotection steps successfully yielded two types of complex-type triantennary oligosaccharides. 26926155 4 63 theme biantennary 721:731 arg1 oligosaccharide 733:747 a biantennary oligosaccharide 719:747 a biantennary oligosaccharide 719:747 We report a highly efficient strategy to access the representative two types of triantennary oligosaccharides through only 9- or 10-step chemical conversions from a biantennary oligosaccharide, which can be isolated in exceptionally homogeneous form from egg yolk. 28670741 2 0 theme xyloglucan 514:523 arg1 chains 530:535 xyloglucan side chains 514:535 xyloglucan side chains 514:535 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 3 1 theme molecular 788:796 arg1 simulations 798:808 molecular simulations 788:808 molecular simulations 788:808 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 4 2 theme structural 938:947 arg1 architecture 949:960 the structural architecture 934:960 the structural architecture of AtFUT1 934:970 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 5 3 theme positioned 1061:1070 arg1 residue 1084:1090 an appropriately positioned active site residue 1044:1090 an appropriately positioned active site residue as a catalytic base 1044:1110 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 1 4 theme complex 168:174 arg1 process 176:182 the complex process 164:182 the complex process of plant polysaccharide biosynthesis 164:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 5 5 theme detailed 1278:1285 arg1 simulations 1297:1307 detailed atomistic simulations 1278:1307 mutagenesis experiments as well as detailed atomistic simulations 1243:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 0 6 theme Arabidopsis 74:84 arg1 thaliana 86:93 Arabidopsis thaliana 74:93 Arabidopsis thaliana 74:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 3 7 theme multipronged 685:696 arg1 approach 698:705 a multipronged approach 683:705 a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations 683:808 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 1 8 theme process 176:182 arg1 underpinnings 147:159 The mechanistic underpinnings 131:159 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis 131:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 5 9 theme atomistic 1287:1295 arg1 simulations 1297:1307 detailed atomistic simulations 1278:1307 mutagenesis experiments as well as detailed atomistic simulations 1243:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 5 10 theme catalytic 1097:1105 arg1 base 1107:1110 a catalytic base 1095:1110 a catalytic base 1095:1110 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 4 11 theme culture 826:832 arg1 expressions 834:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 1 12 theme glycosyltransferase 281:299 arg1 enzymes 306:312 glycosyltransferase (GT) enzymes 281:312 glycosyltransferase (GT) enzymes 281:312 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 2 13 theme key 541:543 arg1 step 545:548 a key step 539:548 a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan 539:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 4 14 theme cell 821:824 arg1 expressions 834:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 2 15 theme transferase 382:392 arg1 fucosyltransferase 411:428 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 371:430 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) 371:439 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 15 theme transferase 382:392 arg1 AtFUT1 433:438 AtFUT1 433:438 AtFUT1 433:438 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 16 theme GT37 405:408 arg1 fucosyltransferase 411:428 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 371:430 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) 371:439 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 16 theme GT37 405:408 arg1 AtFUT1 433:438 AtFUT1 433:438 AtFUT1 433:438 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 5 17 theme fucosylation 1159:1170 arg1 mechanism 1172:1180 an atypical water-mediated fucosylation mechanism 1132:1180 an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations 1132:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 0 18 theme water-mediated 105:118 arg1 mechanism 120:128 a water-mediated mechanism 103:128 a water-mediated mechanism 103:128 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 0 19 from studies 36:42 arg1 thaliana 86:93 Arabidopsis thaliana 74:93 Arabidopsis thaliana 74:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 5 20 theme H-bonded 1200:1207 arg1 network 1209:1215 an H-bonded network 1197:1215 an H-bonded network 1197:1215 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 2 21 theme regiospecific 455:467 arg1 transfer 469:476 the regiospecific transfer 451:476 the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains 451:535 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 1 22 theme plant 187:191 arg1 biosynthesis 208:219 plant polysaccharide biosynthesis 187:219 plant polysaccharide biosynthesis 187:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 3 23 theme mutagenesis 760:770 arg1 experiments 772:782 mutagenesis experiments 760:782 mutagenesis experiments 760:782 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 1 24 theme polysaccharide 193:206 arg1 biosynthesis 208:219 plant polysaccharide biosynthesis 187:219 plant polysaccharide biosynthesis 187:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 0 25 theme Structural 0:9 arg1 studies 36:42 Structural, mutagenic and in silico studies 0:42 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana 0:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 1 26 theme GT 302:303 arg1 enzymes 306:312 glycosyltransferase (GT) enzymes 281:312 glycosyltransferase (GT) enzymes 281:312 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 1 27 theme biosynthesis 208:219 arg1 process 176:182 the complex process 164:182 the complex process of plant polysaccharide biosynthesis 164:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 5 28 theme site 1079:1082 arg1 residue 1084:1090 an appropriately positioned active site residue 1044:1090 an appropriately positioned active site residue as a catalytic base 1044:1110 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 4 29 theme donor 994:998 arg1 analogs 1023:1029 bound donor and acceptor substrate analogs 988:1029 bound donor and acceptor substrate analogs 988:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 4 30 theme Mammalian 811:819 arg1 expressions 834:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions 811:844 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 3 31 with AtFUT1 671:676 arg1 approach 698:705 a multipronged approach 683:705 a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations 683:808 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 0 32 theme mutagenic 12:20 arg1 studies 36:42 Structural, mutagenic and in silico studies 0:42 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana 0:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 1 33 theme enzymes 306:312 arg1 resistance 267:276 the resistance 263:276 the resistance of glycosyltransferase (GT) enzymes to structural characterization 263:343 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 4 34 theme bound 988:992 arg1 analogs 1023:1029 bound donor and acceptor substrate analogs 988:1029 bound donor and acceptor substrate analogs 988:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 2 35 theme glycosyl 373:380 arg1 fucosyltransferase 411:428 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 371:430 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) 371:439 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 35 theme glycosyl 373:380 arg1 AtFUT1 433:438 AtFUT1 433:438 AtFUT1 433:438 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 36 theme terminal 481:488 arg1 residues 502:509 terminal 1,2-fucosyl residues 481:509 terminal 1,2-fucosyl residues 481:509 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 37 theme residues 502:509 arg1 transfer 469:476 the regiospecific transfer 451:476 the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains 451:535 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 4 38 theme acceptor 1004:1011 arg1 analogs 1023:1029 bound donor and acceptor substrate analogs 988:1029 bound donor and acceptor substrate analogs 988:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 4 39 with complex 975:981 arg1 analogs 1023:1029 bound donor and acceptor substrate analogs 988:1029 bound donor and acceptor substrate analogs 988:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 4 40 theme enzyme 886:891 arg1 production 868:877 the sufficient production 853:877 the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs 853:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 0 41 theme in silico 26:34 arg1 studies 36:42 Structural, mutagenic and in silico studies 0:42 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana 0:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 1 42 theme structural 317:326 arg1 characterization 328:343 structural characterization 317:343 structural characterization 317:343 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 2 43 theme 1,2-fucosyl 490:500 arg1 residues 502:509 terminal 1,2-fucosyl residues 481:509 terminal 1,2-fucosyl residues 481:509 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 5 44 theme water-mediated 1144:1157 arg1 mechanism 1172:1180 an atypical water-mediated fucosylation mechanism 1132:1180 an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations 1132:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 0 45 theme xyloglucan 47:56 arg1 fucosylation 58:69 xyloglucan fucosylation 47:69 xyloglucan fucosylation 47:69 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 2 46 theme galactoxyloglucan 599:615 arg1 sidechains 585:594 fucosylated sidechains 573:594 fucosylated sidechains of galactoxyloglucan 573:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 47 theme side 525:528 arg1 chains 530:535 xyloglucan side chains 514:535 xyloglucan side chains 514:535 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 4 48 theme X-ray 897:901 arg1 crystallography 903:917 X-ray crystallography 897:917 X-ray crystallography 897:917 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 4 49 theme AtFUT1 965:970 arg1 architecture 949:960 the structural architecture 934:960 the structural architecture of AtFUT1 934:970 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 5 50 theme mutagenesis 1243:1253 arg1 experiments 1255:1265 mutagenesis experiments 1243:1265 mutagenesis experiments as well as detailed atomistic simulations 1243:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 2 51 theme sidechains 585:594 arg1 biosynthesis 557:568 the biosynthesis 553:568 the biosynthesis of fucosylated sidechains of galactoxyloglucan 553:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 3 52 theme protein 717:723 arg1 expression 725:734 protein expression 717:734 protein expression 717:734 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 4 53 theme sufficient 857:866 arg1 production 868:877 the sufficient production 853:877 the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs 853:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 28670741 2 54 gly fucosylated 573:583 arg1 sidechains 585:594 fucosylated sidechains 573:594 fucosylated sidechains of galactoxyloglucan 573:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 55 theme fucosylated 573:583 arg1 sidechains 585:594 fucosylated sidechains 573:594 fucosylated sidechains of galactoxyloglucan 573:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 56 from step 545:548 arg1 biosynthesis 557:568 the biosynthesis 553:568 the biosynthesis of fucosylated sidechains of galactoxyloglucan 553:615 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 5 57 theme active 1072:1077 arg1 residue 1084:1090 an appropriately positioned active site residue 1044:1090 an appropriately positioned active site residue as a catalytic base 1044:1110 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 5 58 theme atypical 1135:1142 arg1 mechanism 1172:1180 an atypical water-mediated fucosylation mechanism 1132:1180 an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations 1132:1307 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 2 59 theme Arabidopsis 349:359 arg1 thaliana 361:368 Arabidopsis thaliana 349:368 Arabidopsis thaliana 349:368 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 1 60 theme mechanistic 135:145 arg1 underpinnings 147:159 The mechanistic underpinnings 131:159 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis 131:219 The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. 28670741 3 61 theme mechanistic 633:643 arg1 basis 645:649 the mechanistic basis 629:649 the mechanistic basis for fucosylation 629:666 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 0 62 theme fucosylation 58:69 arg1 studies 36:42 Structural, mutagenic and in silico studies 0:42 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana 0:93 Structural, mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water-mediated mechanism. 28670741 3 63 theme X-ray 737:741 arg1 crystallography 743:757 X-ray crystallography 737:757 X-ray crystallography 737:757 We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X-ray crystallography, mutagenesis experiments and molecular simulations. 28670741 5 64 theme residue 1084:1090 arg1 lack 1036:1039 The lack 1032:1039 The lack of an appropriately positioned active site residue as a catalytic base 1032:1110 The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water-mediated fucosylation mechanism facilitated by an H-bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations. 28670741 2 65 theme family 394:399 arg1 fucosyltransferase 411:428 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 371:430 a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) 371:439 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 2 65 theme family 394:399 arg1 AtFUT1 433:438 AtFUT1 433:438 AtFUT1 433:438 In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2-fucosyl residues to xyloglucan side chains - a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. 28670741 4 66 theme substrate 1013:1021 arg1 analogs 1023:1029 bound donor and acceptor substrate analogs 988:1029 bound donor and acceptor substrate analogs 988:1029 Mammalian cell culture expressions enable the sufficient production of the enzyme for X-ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. 29245137 0 0 theme water-borne 79:89 arg1 bacteria 91:98 the water-borne bacteria 75:98 the water-borne bacteria 75:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 5 1 theme linear 706:711 arg1 steps 713:717 11 linear steps 703:717 11 linear steps from the monosaccharide building blocks 10 and 14 703:767 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 1 2 theme bacterial 154:162 arg1 bestiarum 135:143 Aeromonas bestiarum 207 125:147 Aeromonas bestiarum 207 125:147 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 1 2 theme bacterial 154:162 arg1 pathogen 164:171 a bacterial pathogen 152:171 a bacterial pathogen with severe impact on aquaculture 152:205 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 1 3 with pathogen 164:171 arg1 impact 185:190 severe impact 178:190 severe impact on aquaculture 178:205 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 3 4 theme repeating 366:374 arg1 unit 376:379 the pentasaccharide repeating unit 346:379 the pentasaccharide repeating unit 346:379 Synthesis of the pentasaccharide repeating unit and its derivative are reported. 29245137 4 5 theme building 580:587 arg1 blocks 589:594 appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks 513:594 blocks 589:594 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 5 6 theme overall 673:679 arg1 yield 681:685 an overall yield 670:685 an overall yield of 5.2% 670:693 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 0 7 dep 207 120:122 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria 0:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 3 8 theme unit 376:379 arg1 Synthesis 333:341 Synthesis 333:341 Synthesis of the pentasaccharide repeating unit 333:379 Synthesis of the pentasaccharide repeating unit and its derivative are reported. 29245137 3 8 theme unit 376:379 arg1 derivative 389:398 its derivative 385:398 its derivative 385:398 Synthesis of the pentasaccharide repeating unit and its derivative are reported. 29245137 3 9 theme pentasaccharide 350:364 arg1 unit 376:379 the pentasaccharide repeating unit 346:379 the pentasaccharide repeating unit 346:379 Synthesis of the pentasaccharide repeating unit and its derivative are reported. 29245137 5 10 theme % 693:693 arg1 yield 681:685 an overall yield 670:685 an overall yield of 5.2% 670:693 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 5 11 theme [3 + 2 648:653 arg1 strategy 656:663 a [3 + 2] strategy 646:663 a [3 + 2] strategy 646:663 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 1 12 theme severe 178:183 arg1 impact 185:190 severe impact 178:190 severe impact on aquaculture 178:205 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 4 13 theme protected 527:535 arg1 L-rhamopyranosyl 537:552 appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks 513:594 L-rhamopyranosyl 537:552 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 2 14 theme repeating 316:324 arg1 units 326:330 pentasaccharide repeating units 300:330 pentasaccharide repeating units 300:330 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 14 theme repeating 316:324 arg1 structure 231:239 the structure 227:239 the structure of OPS antigens from Aeromonas bestiarum 227:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 15 theme pentasaccharide 300:314 arg1 units 326:330 pentasaccharide repeating units 300:330 pentasaccharide repeating units 300:330 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 15 theme pentasaccharide 300:314 arg1 structure 231:239 the structure 227:239 the structure of OPS antigens from Aeromonas bestiarum 227:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 0 16 theme repeating 17:25 arg1 unit 27:30 the repeating unit 13:30 the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria 13:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 2 17 from structure 231:239 arg1 bestiarum 272:280 Aeromonas bestiarum 262:280 Aeromonas bestiarum 262:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 4 18 theme Schmidt 470:476 arg1 conditions 492:501 Schmidt glycosylation conditions 470:501 Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks 470:594 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 0 19 theme unit 27:30 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria 0:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 0 20 attach isolated 61:68 arg2 polysaccharide 46:59 O-specific polysaccharide 35:59 O-specific polysaccharide isolated from the water-borne bacteria 35:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 0 20 attach isolated 61:68 arg1 bacteria 91:98 the water-borne bacteria 75:98 the water-borne bacteria 75:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 0 21 theme polysaccharide 46:59 arg1 unit 27:30 the repeating unit 13:30 the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria 13:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 5 22 dep blocks 752:757 arg1 14 766:767 14 766:767 14 766:767 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 5 22 dep blocks 752:757 arg1 10 759:760 10 759:760 10 759:760 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 5 23 theme monosaccharide 728:741 arg1 blocks 752:757 the monosaccharide building blocks 10 and 14 724:767 the monosaccharide building blocks 10 and 14 724:767 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 4 24 theme glycosylation 478:490 arg1 conditions 492:501 Schmidt glycosylation conditions 470:501 Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks 470:594 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 0 25 theme O-specific 35:44 arg1 polysaccharide 46:59 O-specific polysaccharide 35:59 O-specific polysaccharide isolated from the water-borne bacteria 35:98 Synthesis of the repeating unit of O-specific polysaccharide isolated from the water-borne bacteria Aeromonas bestiarum 207. 29245137 5 26 theme building 743:750 arg1 blocks 752:757 the monosaccharide building blocks 10 and 14 724:767 the monosaccharide building blocks 10 and 14 724:767 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 2 27 theme recent 213:218 arg1 study 220:224 a recent study 211:224 a recent study 211:224 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 1 28 theme Aeromonas 125:133 arg1 bestiarum 135:143 Aeromonas bestiarum 207 125:147 Aeromonas bestiarum 207 125:147 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 1 28 theme Aeromonas 125:133 arg1 pathogen 164:171 a bacterial pathogen 152:171 a bacterial pathogen with severe impact on aquaculture 152:205 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 4 29 theme regio-specific 426:439 arg1 synthesis 441:449 regio-specific synthesis 426:449 regio-specific synthesis 426:449 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 2 30 theme antigens 248:255 arg1 units 326:330 pentasaccharide repeating units 300:330 pentasaccharide repeating units 300:330 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 30 theme antigens 248:255 arg1 structure 231:239 the structure 227:239 the structure of OPS antigens from Aeromonas bestiarum 227:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 5 31 from blocks 752:757 arg1 steps 713:717 11 linear steps 703:717 11 linear steps from the monosaccharide building blocks 10 and 14 703:767 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 29245137 2 32 theme OPS 244:246 arg1 antigens 248:255 OPS antigens 244:255 OPS antigens from Aeromonas bestiarum 244:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 1 33 from impact 185:190 arg1 aquaculture 195:205 aquaculture 195:205 aquaculture 195:205 Aeromonas bestiarum 207 is a bacterial pathogen with severe impact on aquaculture. 29245137 2 34 from bestiarum 272:280 arg1 units 326:330 pentasaccharide repeating units 300:330 pentasaccharide repeating units 300:330 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 34 from bestiarum 272:280 arg1 structure 231:239 the structure 227:239 the structure of OPS antigens from Aeromonas bestiarum 227:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 2 34 from bestiarum 272:280 arg1 antigens 248:255 OPS antigens 244:255 OPS antigens from Aeromonas bestiarum 244:280 In a recent study, the structure of OPS antigens from Aeromonas bestiarum was identified as pentasaccharide repeating units. 29245137 4 35 theme D-glucopyranosylamine 558:578 arg1 blocks 589:594 appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks 513:594 blocks 589:594 Stereo- and regio-specific synthesis was achieved under Schmidt glycosylation conditions employing appropriately protected L-rhamopyranosyl and D-glucopyranosylamine building blocks. 29245137 5 36 theme pentasaccharide 601:615 arg1 synthesis 617:625 The pentasaccharide synthesis 597:625 The pentasaccharide synthesis 597:625 The pentasaccharide synthesis was achieved using a [3 + 2] strategy with an overall yield of 5.2% through 11 linear steps from the monosaccharide building blocks 10 and 14. 28135419 4 0 theme oligosaccharide 516:530 arg1 synthesis 532:540 the expeditious oligosaccharide synthesis 500:540 the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies 500:603 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 2 1 theme active-latent 357:369 arg1 glycosylations 371:384 active-latent glycosylations 357:384 active-latent glycosylations 357:384 The SBiz imidates have been originally developed as a new platform for active-latent glycosylations. 28135419 5 2 theme SBiz 751:754 arg1 imidates 756:763 SBiz imidates 751:763 SBiz imidates 751:763 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 4 3 theme selective 552:560 arg1 strategies 594:603 selective, chemoselective, and orthogonal strategies 552:603 selective, chemoselective, and orthogonal strategies 552:603 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 5 4 theme reaction 709:716 arg1 mechanism 718:726 the reaction mechanism 705:726 the reaction mechanism 705:726 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 1 5 theme oligosaccharide 259:273 arg1 synthesis 275:283 oligosaccharide synthesis 259:283 oligosaccharide synthesis 259:283 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 0 6 theme Active-Latent 81:93 arg1 Activations 138:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 S-Benzimidazolyl (SBiz) Imidates as a Platform for Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations. 28135419 5 7 theme mechanism 718:726 arg1 modes 736:740 the modes 732:740 the modes by which SBiz imidates interact with various promoters of glycosylation 732:812 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 5 7 theme mechanism 718:726 arg1 understanding 688:700 our enhanced understanding 675:700 our enhanced understanding of the reaction mechanism 675:726 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 2 8 theme SBiz 290:293 arg1 imidates 295:302 The SBiz imidates 286:302 The SBiz imidates 286:302 The SBiz imidates have been originally developed as a new platform for active-latent glycosylations. 28135419 2 8 theme SBiz 290:293 arg1 platform 344:351 a new platform 338:351 a new platform for active-latent glycosylations 338:384 The SBiz imidates have been originally developed as a new platform for active-latent glycosylations. 28135419 0 9 theme Armed-Disarmed 96:109 arg1 Activations 138:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 S-Benzimidazolyl (SBiz) Imidates as a Platform for Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations. 28135419 4 10 theme expeditious 504:514 arg1 synthesis 532:540 the expeditious oligosaccharide synthesis 500:540 the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies 500:603 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 5 11 theme enhanced 679:686 arg1 understanding 688:700 our enhanced understanding 675:700 our enhanced understanding of the reaction mechanism 675:726 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 5 12 theme glycosylation 800:812 arg1 promoters 787:795 various promoters 779:795 various promoters of glycosylation 779:812 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 3 13 theme compounds 434:442 arg1 utility 417:423 the utility 413:423 the utility of these compounds 413:442 This article expands upon the utility of these compounds. 28135419 5 14 theme strategy 626:633 arg1 development 635:645 The strategy development 622:645 The strategy development 622:645 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 4 15 theme orthogonal 583:592 arg1 strategies 594:603 selective, chemoselective, and orthogonal strategies 552:603 selective, chemoselective, and orthogonal strategies 552:603 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 1 16 theme S-benzimidazolyl 193:208 arg1 imidates 217:224 S-benzimidazolyl (SBiz) imidates 193:224 S-benzimidazolyl (SBiz) imidates 193:224 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 0 17 theme Selective 112:120 arg1 Activations 138:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 S-Benzimidazolyl (SBiz) Imidates as a Platform for Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations. 28135419 0 18 theme Orthogonal 127:136 arg1 Activations 138:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 81:148 S-Benzimidazolyl (SBiz) Imidates as a Platform for Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations. 28135419 1 19 theme SBiz 211:214 arg1 imidates 217:224 S-benzimidazolyl (SBiz) imidates 193:224 S-benzimidazolyl (SBiz) imidates 193:224 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 4 20 theme chemoselective 563:576 arg1 strategies 594:603 selective, chemoselective, and orthogonal strategies 552:603 selective, chemoselective, and orthogonal strategies 552:603 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 4 21 theme common 480:485 arg1 concepts 487:494 practically all common concepts 464:494 practically all common concepts 464:494 The application to practically all common concepts for the expeditious oligosaccharide synthesis including selective, chemoselective, and orthogonal strategies is demonstrated. 28135419 2 22 theme new 340:342 arg1 imidates 295:302 The SBiz imidates 286:302 The SBiz imidates 286:302 The SBiz imidates have been originally developed as a new platform for active-latent glycosylations. 28135419 2 22 theme new 340:342 arg1 platform 344:351 a new platform 338:351 a new platform for active-latent glycosylations 338:384 The SBiz imidates have been originally developed as a new platform for active-latent glycosylations. 28135419 5 23 dep possible 656:663 arg1 thanks 665:670 thanks 665:670 thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation 665:812 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 1 24 theme imidates 217:224 arg1 development 178:188 the development 174:188 the development of S-benzimidazolyl (SBiz) imidates 174:224 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 1 24 theme imidates 217:224 arg1 blocks 248:253 versatile building blocks 229:253 versatile building blocks for oligosaccharide synthesis 229:283 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 5 25 theme various 779:785 arg1 promoters 787:795 various promoters 779:795 various promoters of glycosylation 779:812 The strategy development was made possible thanks to our enhanced understanding of the reaction mechanism and the modes by which SBiz imidates interact with various promoters of glycosylation. 28135419 0 26 theme Oligosaccharide 51:65 arg1 Synthesis 67:75 Oligosaccharide Synthesis 51:75 Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations 51:148 S-Benzimidazolyl (SBiz) Imidates as a Platform for Oligosaccharide Synthesis via Active-Latent, Armed-Disarmed, Selective, and Orthogonal Activations. 28135419 1 27 theme versatile 229:237 arg1 development 178:188 the development 174:188 the development of S-benzimidazolyl (SBiz) imidates 174:224 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 1 27 theme versatile 229:237 arg1 blocks 248:253 versatile building blocks 229:253 versatile building blocks for oligosaccharide synthesis 229:283 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 1 28 theme building 239:246 arg1 development 178:188 the development 174:188 the development of S-benzimidazolyl (SBiz) imidates 174:224 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 28135419 1 28 theme building 239:246 arg1 blocks 248:253 versatile building blocks 229:253 versatile building blocks for oligosaccharide synthesis 229:283 This article describes the development of S-benzimidazolyl (SBiz) imidates as versatile building blocks for oligosaccharide synthesis. 26112419 0 0 theme EndoS 75:79 arg1 deglycosylation 81:95 EndoS deglycosylation 75:95 EndoS deglycosylation 75:95 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 4 1 theme EndoS 664:668 arg1 deglycosylation 670:684 EndoS deglycosylation 664:684 EndoS deglycosylation 664:684 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 5 2 theme wider 1024:1028 arg1 variety 1030:1036 a wider variety 1022:1036 a wider variety of structures upon deglycosylation 1022:1071 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 5 2 theme wider 1024:1028 arg1 structures 1041:1050 structures 1041:1050 structures upon deglycosylation 1041:1071 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 5 3 gly deglycosylated 955:968 arg1 antibodies 970:979 deglycosylated antibodies 955:979 deglycosylated antibodies 955:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 4 4 theme dye-labeled 830:840 arg1 antibodies 860:869 dye-labeled, freely diffusing antibodies 830:869 dye-labeled, freely diffusing antibodies 830:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 5 theme region 718:723 arg1 structure 698:706 the structure 694:706 the structure of the Fc region of IgG antibodies 694:741 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 6 6 from presence 1102:1109 arg1 histogram 1154:1162 the FRET efficiency histogram 1134:1162 the FRET efficiency histogram for the deglycosylated case 1134:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 3 7 theme specific 498:505 arg1 nature 507:512 the specific nature 494:512 the specific nature of this structural change 494:538 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 2 8 theme immune 282:287 arg1 response 289:296 a diminished immune response 269:296 a diminished immune response 269:296 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 0 9 theme deglycosylation 81:95 arg1 effect 65:70 the effect 61:70 the effect of EndoS deglycosylation on the structure of IgG 61:119 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 3 10 theme change 533:538 arg1 nature 507:512 the specific nature 494:512 the specific nature of this structural change 494:538 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 2 11 theme diminished 271:280 arg1 response 289:296 a diminished immune response 269:296 a diminished immune response 269:296 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 6 12 theme efficiency 1143:1152 arg1 histogram 1154:1162 the FRET efficiency histogram 1134:1162 the FRET efficiency histogram for the deglycosylated case 1134:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 0 13 from studies 50:56 arg1 structure 104:112 the structure 100:112 the structure of IgG 100:119 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 3 14 theme Fc 465:466 arg1 region 468:473 the Fc region 461:473 the Fc region of IgG antibodies 461:491 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 5 15 theme deglycosylated 955:968 arg1 antibodies 970:979 deglycosylated antibodies 955:979 deglycosylated antibodies 955:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 4 16 dep dye-labeled 830:840 arg1 diffusing 850:858 diffusing 850:858 diffusing 850:858 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 17 theme Förster 778:784 arg1 FRET 813:816 FRET 813:816 FRET 813:816 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 17 theme Förster 778:784 arg1 transfer 803:810 single molecule Förster resonance energy transfer 762:810 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 5 18 theme histograms 908:917 arg1 comparison 874:883 A comparison 872:883 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies 872:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 3 19 theme structural 440:449 arg1 change 451:456 a structural change 438:456 a structural change in the Fc region of IgG antibodies 438:491 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 0 20 theme molecule 7:14 arg1 transfer 41:48 Single molecule Förster resonance energy transfer 0:48 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 6 21 theme FRET 1138:1141 arg1 histogram 1154:1162 the FRET efficiency histogram 1134:1162 the FRET efficiency histogram for the deglycosylated case 1134:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 0 22 gly deglycosylation 81:95 arg1 IgG 117:119 IgG 117:119 IgG 117:119 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 0 23 theme Single 0:5 arg1 transfer 41:48 Single molecule Förster resonance energy transfer 0:48 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 4 24 theme Fc 715:716 arg1 region 718:723 the Fc region 711:723 the Fc region of IgG antibodies 711:741 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 5 25 dep glycosylated 932:943 arg1 antibodies 970:979 deglycosylated antibodies 955:979 deglycosylated antibodies 955:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 0 26 theme resonance 24:32 arg1 transfer 41:48 Single molecule Förster resonance energy transfer 0:48 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 3 27 theme results 575:581 arg1 variety 564:570 the variety 560:570 the variety of results obtained by different experimental approaches 560:627 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 3 27 theme results 575:581 arg1 results 575:581 results 575:581 results obtained by different experimental approaches 575:627 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 0 28 theme IgG 117:119 arg1 structure 104:112 the structure 100:112 the structure of IgG 100:119 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 4 29 theme IgG 728:730 arg1 antibodies 732:741 IgG antibodies 728:741 IgG antibodies 728:741 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 6 30 from peaks 1125:1129 arg1 histogram 1154:1162 the FRET efficiency histogram 1134:1162 the FRET efficiency histogram for the deglycosylated case 1134:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 1 31 theme immunoglobulin 187:200 arg1 G 202:202 immunoglobulin G 187:202 immunoglobulin G (IgG) molecules 187:218 The bacterial enzyme EndoS specifically cleaves glycans bound to immunoglobulin G (IgG) molecules. 26112419 1 31 theme immunoglobulin 187:200 arg1 IgG 205:207 IgG 205:207 IgG 205:207 The bacterial enzyme EndoS specifically cleaves glycans bound to immunoglobulin G (IgG) molecules. 26112419 0 32 theme Förster 16:22 arg1 transfer 41:48 Single molecule Förster resonance energy transfer 0:48 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 5 33 theme structures 1041:1050 arg1 variety 1030:1036 a wider variety 1022:1036 a wider variety of structures upon deglycosylation 1022:1071 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 5 33 theme structures 1041:1050 arg1 structures 1041:1050 structures 1041:1050 structures upon deglycosylation 1041:1071 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 6 34 theme peaks 1125:1129 arg1 presence 1102:1109 the presence 1098:1109 the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case 1098:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 3 35 theme experimental 605:616 arg1 approaches 618:627 different experimental approaches 595:627 different experimental approaches 595:627 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 1 36 theme G 202:202 arg1 molecules 210:218 immunoglobulin G (IgG) molecules 187:218 immunoglobulin G (IgG) molecules 187:218 The bacterial enzyme EndoS specifically cleaves glycans bound to immunoglobulin G (IgG) molecules. 26112419 0 37 theme transfer 41:48 arg1 studies 50:56 Single molecule Förster resonance energy transfer studies 0:56 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 6 38 from histogram 1154:1162 arg1 presence 1102:1109 the presence 1098:1109 the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case 1098:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 4 39 theme energy 796:801 arg1 FRET 813:816 FRET 813:816 FRET 813:816 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 39 theme energy 796:801 arg1 transfer 803:810 single molecule Förster resonance energy transfer 762:810 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 6 40 theme deglycosylated 1172:1185 arg1 case 1187:1190 the deglycosylated case 1168:1190 the deglycosylated case 1168:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 0 41 theme energy 34:39 arg1 transfer 41:48 Single molecule Förster resonance energy transfer 0:48 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 6 42 gly deglycosylated 1172:1185 arg1 case 1187:1190 the deglycosylated case 1168:1190 the deglycosylated case 1168:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 0 43 from effect 65:70 arg1 structure 104:112 the structure 100:112 the structure of IgG 100:119 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 4 44 theme resonance 786:794 arg1 FRET 813:816 FRET 813:816 FRET 813:816 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 44 theme resonance 786:794 arg1 transfer 803:810 single molecule Förster resonance energy transfer 762:810 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 3 45 from change 451:456 arg1 region 468:473 the Fc region 461:473 the Fc region of IgG antibodies 461:491 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 3 46 theme diminished 394:403 arg1 response 412:419 the diminished immune response 390:419 the diminished immune response 390:419 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 4 47 theme antibodies 732:741 arg1 region 718:723 the Fc region 711:723 the Fc region of IgG antibodies 711:741 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 3 48 theme IgG 478:480 arg1 antibodies 482:491 IgG antibodies 478:491 IgG antibodies 478:491 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 3 49 theme immune 405:410 arg1 response 412:419 the diminished immune response 390:419 the diminished immune response 390:419 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 4 50 theme antibodies 860:869 arg1 studies 819:825 single molecule Förster resonance energy transfer (FRET) studies 762:825 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 0 51 from structure 104:112 arg1 studies 50:56 Single molecule Förster resonance energy transfer studies 0:56 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 4 52 theme transfer 803:810 arg1 studies 819:825 single molecule Förster resonance energy transfer (FRET) studies 762:825 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 3 53 theme antibodies 482:491 arg1 region 468:473 the Fc region 461:473 the Fc region of IgG antibodies 461:491 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 5 54 theme Fc 1000:1001 arg1 region 1003:1008 the Fc region 996:1008 the Fc region 996:1008 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 1 55 theme bacterial 126:134 arg1 EndoS 143:147 The bacterial enzyme EndoS 122:147 The bacterial enzyme EndoS 122:147 The bacterial enzyme EndoS specifically cleaves glycans bound to immunoglobulin G (IgG) molecules. 26112419 4 56 theme single 762:767 arg1 FRET 813:816 FRET 813:816 FRET 813:816 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 56 theme single 762:767 arg1 transfer 803:810 single molecule Förster resonance energy transfer 762:810 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 2 57 theme potential 315:323 arg1 applications 325:336 potential applications 315:336 potential applications 315:336 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 2 57 theme potential 315:323 arg1 therapeutic 343:353 therapeutic 343:353 therapeutic 343:353 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 4 58 dep understand 649:658 arg1 to 639:640 to 639:640 to 639:640 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 3 59 theme different 595:603 arg1 approaches 618:627 different experimental approaches 595:627 different experimental approaches 595:627 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 5 60 theme efficiency 897:906 arg1 histograms 908:917 the FRET efficiency histograms 888:917 the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies 888:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 6 61 theme additional 1114:1123 arg1 peaks 1125:1129 additional peaks 1114:1129 additional peaks in the FRET efficiency histogram for the deglycosylated case 1114:1190 This is demonstrated by the presence of additional peaks in the FRET efficiency histogram for the deglycosylated case. 26112419 5 62 theme FRET 892:895 arg1 histograms 908:917 the FRET efficiency histograms 888:917 the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies 888:979 A comparison of the FRET efficiency histograms obtained for glycosylated and EndoS deglycosylated antibodies indicates that the Fc region can take on a wider variety of structures upon deglycosylation. 26112419 0 63 theme effect 65:70 arg1 studies 50:56 Single molecule Förster resonance energy transfer studies 0:56 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 0:120 Single molecule Förster resonance energy transfer studies of the effect of EndoS deglycosylation on the structure of IgG. 26112419 2 64 theme deglycosylation 234:248 arg1 procedure 250:258 this deglycosylation procedure 229:258 this deglycosylation procedure 229:258 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 3 65 theme structural 522:531 arg1 change 533:538 this structural change 517:538 this structural change 517:538 Although the diminished immune response is attributed to a structural change in the Fc region of IgG antibodies, the specific nature of this structural change is not known due to the variety of results obtained by different experimental approaches. 26112419 2 66 theme autoimmune 359:368 arg1 disorders 370:378 autoimmune disorders 359:378 autoimmune disorders 359:378 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 2 67 contain has 311:313 arg2 therapeutic 343:353 therapeutic 343:353 therapeutic 343:353 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 2 67 contain has 311:313 arg2 applications 325:336 potential applications 315:336 potential applications 315:336 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 2 67 contain has 311:313 arg1 enzyme 304:309 this enzyme 299:309 this enzyme 299:309 Because this deglycosylation procedure leads to a diminished immune response, this enzyme has potential applications as a therapeutic for autoimmune disorders. 26112419 4 68 theme molecule 769:776 arg1 FRET 813:816 FRET 813:816 FRET 813:816 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 4 68 theme molecule 769:776 arg1 transfer 803:810 single molecule Förster resonance energy transfer 762:810 single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies 762:869 In order to better understand how EndoS deglycosylation impacts the structure of the Fc region of IgG antibodies, we have conducted single molecule Förster resonance energy transfer (FRET) studies of dye-labeled, freely diffusing antibodies. 26112419 1 69 theme enzyme 136:141 arg1 EndoS 143:147 The bacterial enzyme EndoS 122:147 The bacterial enzyme EndoS 122:147 The bacterial enzyme EndoS specifically cleaves glycans bound to immunoglobulin G (IgG) molecules. 24785386 4 0 theme 3+2 488:490 arg1 strategy 507:514 a convergent '3+2' glycosylation strategy 474:514 a convergent '3+2' glycosylation strategy 474:514 APK2, and was synthesized in high overall yield via a convergent '3+2' glycosylation strategy. 24785386 2 1 theme α-l-fucopyranosyl- 115:132 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 4 2 theme high 451:454 arg1 yield 464:468 high overall yield 451:468 high overall yield 451:468 APK2, and was synthesized in high overall yield via a convergent '3+2' glycosylation strategy. 24785386 3 3 theme polysaccharide 356:369 arg1 unit 344:347 the repeating unit 330:347 the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var 330:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 2 4 theme p-methoxyphenyl 99:113 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 3 5 dep mushroom 392:399 arg1 var 417:419 Calocybe indica var 401:419 edible mushroom Calocybe indica var 385:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 2 6 theme building 287:294 arg1 blocks 296:301 five monosaccharide building blocks 267:301 five monosaccharide building blocks 267:301 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 2 7 theme 1→4 161:163 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 2 8 theme monosaccharide 272:285 arg1 blocks 296:301 five monosaccharide building blocks 267:301 five monosaccharide building blocks 267:301 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 2 9 theme -α-d-galactopyranosyl- 138:159 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 4 10 theme convergent 476:485 arg1 strategy 507:514 a convergent '3+2' glycosylation strategy 474:514 a convergent '3+2' glycosylation strategy 474:514 APK2, and was synthesized in high overall yield via a convergent '3+2' glycosylation strategy. 24785386 3 11 theme Calocybe 401:408 arg1 var 417:419 Calocybe indica var 401:419 edible mushroom Calocybe indica var 385:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 0 12 theme indica 22:27 arg1 var 29:31 Calocybe indica var 13:31 Calocybe indica var 13:31 Synthesis of Calocybe indica var. 24785386 0 13 theme Calocybe 13:20 arg1 var 29:31 Calocybe indica var 13:31 Calocybe indica var 13:31 Synthesis of Calocybe indica var. 24785386 2 14 theme 1→6 186:188 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 0 15 theme var 29:31 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Calocybe indica var. 0:32 Synthesis of Calocybe indica var. 24785386 2 16 theme -β-d-glucopyranosyl- 165:184 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 3 17 theme indica 410:415 arg1 var 417:419 Calocybe indica var 401:419 edible mushroom Calocybe indica var 385:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 4 18 theme overall 456:462 arg1 yield 464:468 high overall yield 451:468 high overall yield 451:468 APK2, and was synthesized in high overall yield via a convergent '3+2' glycosylation strategy. 24785386 2 19 theme -β-d-glucopyranoside 215:234 arg1 synthesis 86:94 The first total synthesis 70:94 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 70:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 4 20 theme glycosylation 493:505 arg1 strategy 507:514 a convergent '3+2' glycosylation strategy 474:514 a convergent '3+2' glycosylation strategy 474:514 APK2, and was synthesized in high overall yield via a convergent '3+2' glycosylation strategy. 24785386 2 21 theme total 80:84 arg1 synthesis 86:94 The first total synthesis 70:94 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 70:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 3 22 attach isolated 371:378 arg1 mushroom 392:399 edible mushroom Calocybe indica var 385:419 edible mushroom Calocybe indica var 385:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 3 22 attach isolated 371:378 arg2 polysaccharide 356:369 the polysaccharide 352:369 the polysaccharide isolated from edible mushroom Calocybe indica var 352:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 2 23 theme first 74:78 arg1 synthesis 86:94 The first total synthesis 70:94 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 70:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 2 24 theme -β-d-glucopyranosyl- 190:209 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 3 25 theme repeating 334:342 arg1 unit 344:347 the repeating unit 330:347 the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var 330:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 24785386 1 26 theme APK2 34:37 arg1 polysaccharide 39:52 APK2 polysaccharide 34:52 APK2 polysaccharide 34:52 APK2 polysaccharide repeating unit. 24785386 2 27 theme 1→6 134:136 arg1 -β-d-glucopyranoside 215:234 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) 99:238 The first total synthesis of p-methoxyphenyl α-l-fucopyranosyl-(1→6)-α-d-galactopyranosyl-(1→4)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranosyl-(1→6)-β-d-glucopyranoside (2) was achieved starting from five monosaccharide building blocks. 24785386 3 28 theme edible 385:390 arg1 mushroom 392:399 edible mushroom Calocybe indica var 385:419 edible mushroom Calocybe indica var 385:419 This structure represents the repeating unit of the polysaccharide isolated from edible mushroom Calocybe indica var. 28318221 5 0 theme open 783:786 arg1 form 788:791 an open form 780:791 an open form 780:791 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 6 1 theme hSCT 910:913 arg1 chains 915:920 the two hSCT chains 902:920 the two hSCT chains 902:920 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 6 2 theme double 875:880 arg1 latches 889:895 the double glycan latches 871:895 the double glycan latches from the two hSCT chains 871:920 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 10 3 theme antibodies 1539:1548 arg1 development 1517:1527 the development 1513:1527 the development of better antibodies for therapeutic applications 1513:1577 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 10 3 theme antibodies 1539:1548 arg1 relationship 1410:1421 the relationship 1406:1421 the relationship between the structural stability and effector functions affected by hSCT modification 1406:1507 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 10 4 theme structural 1435:1444 arg1 stability 1446:1454 structural stability 1435:1454 structural stability 1435:1454 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 1 5 theme Fc 170:171 arg1 conformation 173:184 Fc conformation 170:184 Fc conformation 170:184 N-glycosylation on IgG modulates Fc conformation and effector functions. 28318221 10 6 theme hSCT 1491:1494 arg1 modification 1496:1507 hSCT modification 1491:1507 hSCT modification 1491:1507 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 7 7 theme CH2-CH3 1176:1182 arg1 interface 1184:1192 the CH2-CH3 interface 1172:1192 the CH2-CH3 interface 1172:1192 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 5 8 located observed 798:805 arg2 form 788:791 an open form 780:791 an open form 780:791 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 5 8 located observed 798:805 arg1 unit 824:827 an asymmetric unit 810:827 an asymmetric unit in the crystal 810:842 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 5 8 located observed 798:805 arg2 form 771:774 A compact form 761:774 A compact form 761:774 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 7 9 theme hydrogen 1099:1106 arg1 bonds 1108:1112 water-mediated hydrogen bonds 1084:1112 water-mediated hydrogen bonds with the D249-L251 helix 1084:1137 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 3 10 theme effector 601:608 arg1 activity 610:617 gain-of-function effector activity 584:617 gain-of-function effector activity 584:617 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 7 11 theme interface 1184:1192 arg1 region 1162:1167 the pivot region 1152:1167 the pivot region of the CH2-CH3 interface 1152:1192 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 2 12 theme structure 285:293 arg1 glycan 263:268 a human sialo-complex type (hSCT) glycan 229:268 a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 229:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 9 13 theme enhanced 1343:1350 arg1 activities 1361:1370 enhanced effector activities 1343:1370 enhanced effector activities 1343:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 0 14 theme Dependent 104:112 arg1 Cytotoxicity 123:134 the Antibody Dependent Cellular Cytotoxicity 91:134 the Antibody Dependent Cellular Cytotoxicity 91:134 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 3 15 theme Fc 512:513 arg1 receptors 515:523 Fc receptors 512:523 Fc receptors 512:523 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 8 16 theme acid 1236:1239 arg1 modulation 1241:1250 the sialic acid modulation 1225:1250 the sialic acid modulation 1225:1250 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 8 16 theme acid 1236:1239 arg1 exclusive 1268:1276 exclusive 1268:1276 exclusive 1268:1276 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 2 17 theme biantennary 273:283 arg1 structure 285:293 biantennary structure 273:293 biantennary structure 273:293 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 8 18 theme sialic 1229:1234 arg1 modulation 1241:1250 the sialic acid modulation 1225:1250 the sialic acid modulation 1225:1250 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 8 18 theme sialic 1229:1234 arg1 exclusive 1268:1276 exclusive 1268:1276 exclusive 1268:1276 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 2 19 theme cellular 422:429 arg1 ADCC 445:448 ADCC 445:448 ADCC 445:448 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 2 19 theme cellular 422:429 arg1 cytotoxicity 431:442 the antibody dependent cellular cytotoxicity 399:442 the antibody dependent cellular cytotoxicity (ADCC) 399:449 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 8 20 theme double 1199:1204 arg1 latches 1213:1219 The double glycan latches 1195:1219 The double glycan latches 1195:1219 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 8 20 theme double 1199:1204 arg1 exclusive 1268:1276 exclusive 1268:1276 exclusive 1268:1276 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 6 21 theme glycan 882:887 arg1 latches 889:895 the double glycan latches 871:895 the double glycan latches from the two hSCT chains 871:920 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 7 22 theme water-mediated 1084:1097 arg1 bonds 1108:1112 water-mediated hydrogen bonds 1084:1112 water-mediated hydrogen bonds with the D249-L251 helix 1084:1137 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 3 23 from affinity 500:507 arg1 presence 532:539 the presence 528:539 the presence of antigen 528:550 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 2 24 theme dependent 412:420 arg1 ADCC 445:448 ADCC 445:448 ADCC 445:448 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 2 24 theme dependent 412:420 arg1 cytotoxicity 431:442 the antibody dependent cellular cytotoxicity 399:442 the antibody dependent cellular cytotoxicity (ADCC) 399:449 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 5 25 theme asymmetric 813:822 arg1 unit 824:827 an asymmetric unit 810:827 an asymmetric unit in the crystal 810:842 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 5 26 from unit 824:827 arg1 crystal 836:842 the crystal 832:842 the crystal 832:842 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 10 27 theme effector 1460:1467 arg1 functions 1469:1477 effector functions 1460:1477 effector functions 1460:1477 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 1 28 from N-glycosylation 137:151 arg1 IgG 156:158 IgG 156:158 IgG 156:158 N-glycosylation on IgG modulates Fc conformation and effector functions. 28318221 4 29 used used 623:626 arg2 We 620:621 We 620:621 We 620:621 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 3 30 theme antigen 544:550 arg1 presence 532:539 the presence 528:539 the presence of antigen 528:550 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 1 31 theme effector 190:197 arg1 functions 199:207 effector functions 190:207 effector functions 190:207 N-glycosylation on IgG modulates Fc conformation and effector functions. 28318221 0 32 theme Crystal 0:6 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan 0:68 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 9 33 theme crystal 1297:1303 arg1 structure 1305:1313 the first crystal structure 1287:1313 the first crystal structure of glyco-engineered Fc with enhanced effector activities 1287:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 9 33 theme crystal 1297:1303 arg1 This 1279:1282 This 1279:1282 This 1279:1282 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 9 34 theme effector 1352:1359 arg1 activities 1361:1370 enhanced effector activities 1343:1370 enhanced effector activities 1343:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 3 35 theme gain-of-function 584:599 arg1 activity 610:617 gain-of-function effector activity 584:617 gain-of-function effector activity 584:617 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 3 36 theme hSCT 452:455 arg1 modification 457:468 hSCT modification 452:468 hSCT modification 452:468 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 6 37 theme closed 953:958 arg1 conformation 960:971 a closed conformation 951:971 a closed conformation 951:971 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 7 38 dep acid 1017:1020 arg1 NeuNAc 1050:1055 NeuNAc 1050:1055 NeuNAc 1050:1055 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 7 38 dep acid 1017:1020 arg1 acid 1042:1045 N-acetylneuraminic acid 1023:1045 N-acetylneuraminic acid 1023:1045 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 9 39 theme glyco-engineered 1318:1333 arg1 Fc 1335:1336 glyco-engineered Fc 1318:1336 glyco-engineered Fc 1318:1336 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 0 40 theme Cellular 114:121 arg1 Cytotoxicity 123:134 the Antibody Dependent Cellular Cytotoxicity 91:134 the Antibody Dependent Cellular Cytotoxicity 91:134 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 10 41 theme therapeutic 1554:1564 arg1 applications 1566:1577 therapeutic applications 1554:1577 therapeutic applications 1554:1577 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 3 42 theme binding 492:498 arg1 affinity 500:507 the binding affinity 488:507 the binding affinity to Fc receptors in the presence of antigen 488:550 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 0 43 theme IgG-Fc 35:40 arg1 Glycoform 42:50 a Homogeneous IgG-Fc Glycoform 21:50 a Homogeneous IgG-Fc Glycoform 21:50 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 9 44 theme Fc 1335:1336 arg1 structure 1305:1313 the first crystal structure 1287:1313 the first crystal structure of glyco-engineered Fc with enhanced effector activities 1287:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 9 44 theme Fc 1335:1336 arg1 This 1279:1282 This 1279:1282 This 1279:1282 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 1 45 gly N-glycosylation 137:151 arg1 IgG 156:158 IgG 156:158 IgG 156:158 N-glycosylation on IgG modulates Fc conformation and effector functions. 28318221 2 46 theme type 251:254 arg1 glycan 263:268 a human sialo-complex type (hSCT) glycan 229:268 a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 229:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 9 47 with structure 1305:1313 arg1 activities 1361:1370 enhanced effector activities 1343:1370 enhanced effector activities 1343:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 0 48 theme Homogeneous 23:33 arg1 Glycoform 42:50 a Homogeneous IgG-Fc Glycoform 21:50 a Homogeneous IgG-Fc Glycoform 21:50 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 4 49 attach attached 699:706 arg1 domain 720:725 each CH2 domain 711:725 each CH2 domain 711:725 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 4 49 attach attached 699:706 arg2 IgG-Fc 670:675 an IgG-Fc 667:675 an IgG-Fc with homogeneous hSCT attached to each CH2 domain 667:725 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 2 50 theme sialo-complex 237:249 arg1 hSCT 257:260 hSCT 257:260 hSCT 257:260 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 2 50 theme sialo-complex 237:249 arg1 type 251:254 human sialo-complex type 231:254 a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 229:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 7 51 theme D249-L251 1123:1131 arg1 helix 1133:1137 the D249-L251 helix 1119:1137 the D249-L251 helix 1119:1137 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 6 52 theme CH2 936:938 arg1 domains 940:946 the CH2 domains 932:946 the CH2 domains in a closed conformation 932:971 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 10 53 theme better 1532:1537 arg1 antibodies 1539:1548 better antibodies 1532:1548 better antibodies for therapeutic applications 1532:1577 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 6 54 theme compact 852:858 arg1 structure 860:868 the compact structure 848:868 the compact structure 848:868 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 2 55 theme human 231:235 arg1 hSCT 257:260 hSCT 257:260 hSCT 257:260 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 2 55 theme human 231:235 arg1 type 251:254 human sialo-complex type 231:254 a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 229:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 7 56 theme terminal 1001:1008 arg1 residue 1058:1064 the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue 997:1064 the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue 997:1064 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 2 57 theme optimized 358:366 arg1 IgG-Fc 213:218 An IgG-Fc 210:218 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 210:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 2 57 theme optimized 358:366 arg1 glycoform 368:376 an optimized glycoform 355:376 an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC) 355:449 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 0 58 theme Glycoform 42:50 arg1 Structure 8:16 Crystal Structure 0:16 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan 0:68 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 2 59 with glycan 263:268 arg1 α2,6-sialylations 304:320 two α2,6-sialylations 300:320 two α2,6-sialylations 300:320 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 28318221 7 60 theme N-acetylneuraminic 1023:1040 arg1 acid 1042:1045 N-acetylneuraminic acid 1023:1045 N-acetylneuraminic acid 1023:1045 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 7 61 theme sialic 1010:1015 arg1 acid 1017:1020 sialic acid 1010:1020 the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue 997:1064 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 0 62 with Structure 8:16 arg1 N-Glycan 61:68 the N-Glycan 57:68 the N-Glycan 57:68 Crystal Structure of a Homogeneous IgG-Fc Glycoform with the N-Glycan Designed to Maximize the Antibody Dependent Cellular Cytotoxicity. 28318221 9 63 theme first 1291:1295 arg1 structure 1305:1313 the first crystal structure 1287:1313 the first crystal structure of glyco-engineered Fc with enhanced effector activities 1287:1370 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 9 63 theme first 1291:1295 arg1 This 1279:1282 This 1279:1282 This 1279:1282 This is the first crystal structure of glyco-engineered Fc with enhanced effector activities. 28318221 6 64 from domains 940:946 arg1 conformation 960:971 a closed conformation 951:971 a closed conformation 951:971 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 4 65 with IgG-Fc 670:675 arg1 hSCT 694:697 homogeneous hSCT 682:697 homogeneous hSCT 682:697 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 4 66 theme enzymatic 628:636 arg1 glyco-engineering 638:654 enzymatic glyco-engineering 628:654 enzymatic glyco-engineering 628:654 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 7 67 theme acid 1017:1020 arg1 residue 1058:1064 the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue 997:1064 the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue 997:1064 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 3 68 dep enhances 479:486 arg1 provides 575:582 provides 575:582 not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity 470:617 hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen but also in some cases provides gain-of-function effector activity. 28318221 10 69 dep stability 1446:1454 arg1 the 1431:1433 the 1431:1433 the 1431:1433 This work provides insights into the relationship between the structural stability and effector functions affected by hSCT modification and the development of better antibodies for therapeutic applications. 28318221 4 70 theme crystal 742:748 arg1 structure 750:758 its crystal structure 738:758 its crystal structure 738:758 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 7 71 with bonds 1108:1112 arg1 helix 1133:1137 the D249-L251 helix 1119:1137 the D249-L251 helix 1119:1137 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 5 72 theme compact 763:769 arg1 form 771:774 A compact form 761:774 A compact form 761:774 A compact form and an open form were observed in an asymmetric unit in the crystal. 28318221 7 73 theme open 981:984 arg1 structure 986:994 the open structure 977:994 the open structure 977:994 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 4 74 theme CH2 716:718 arg1 domain 720:725 each CH2 domain 711:725 each CH2 domain 711:725 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 4 75 theme homogeneous 682:692 arg1 hSCT 694:697 homogeneous hSCT 682:697 homogeneous hSCT 682:697 We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain and solved its crystal structure. 28318221 8 76 theme glycan 1206:1211 arg1 latches 1213:1219 The double glycan latches 1195:1219 The double glycan latches 1195:1219 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 8 76 theme glycan 1206:1211 arg1 exclusive 1268:1276 exclusive 1268:1276 exclusive 1268:1276 The double glycan latches and the sialic acid modulation may be mutually exclusive. 28318221 7 77 theme pivot 1156:1160 arg1 region 1162:1167 the pivot region 1152:1167 the pivot region of the CH2-CH3 interface 1152:1192 In the open structure, the terminal sialic acid (N-acetylneuraminic acid or NeuNAc) residue interacts through water-mediated hydrogen bonds with the D249-L251 helix, to modulate the pivot region of the CH2-CH3 interface. 28318221 6 78 from chains 915:920 arg1 latches 889:895 the double glycan latches 871:895 the double glycan latches from the two hSCT chains 871:920 In the compact structure, the double glycan latches from the two hSCT chains stabilize the CH2 domains in a closed conformation. 28318221 2 79 dep IgG-Fc 213:218 arg1 contains 220:227 contains 220:227 contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation 220:350 An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). 25753700 2 0 gly glycoproteins 297:309 arg1 glycoproteins 297:309 glycoproteins 297:309 glycoproteins 297:309 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 4 1 theme EUROCarbDB 791:800 arg1 initiative 802:811 the EUROCarbDB initiative 787:811 the EUROCarbDB initiative 787:811 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 4 2 theme databases 715:723 arg1 availability 699:710 the availability 695:710 the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative 695:811 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 3 3 link O-linked 553:560 arg1 glycans 562:568 O-linked glycans 553:568 O-linked glycans 553:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 4 4 theme data 858:861 arg1 collections 863:873 HPLC-glycan data collections 846:873 HPLC-glycan data collections 846:873 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 1 5 theme glycosylation 103:115 arg1 relevance 82:90 The biological relevance 67:90 The biological relevance of protein glycosylation 67:115 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 3 6 theme complete 511:518 arg1 elucidation 531:541 the complete structural elucidation 507:541 the complete structural elucidation of N- and O-linked glycans 507:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 2 7 theme functional 270:279 arg1 relationship 281:292 functional relationship 270:292 functional relationship 270:292 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 2 8 theme methods 401:407 arg1 range 381:385 a range 379:385 a range of analytical methods 379:407 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 4 9 theme collections 863:873 arg1 interpretation 828:841 the interpretation 824:841 the interpretation of HPLC-glycan data collections 824:873 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 1 10 theme biological 71:80 arg1 relevance 82:90 The biological relevance 67:90 The biological relevance of protein glycosylation 67:115 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 2 11 theme comprehensive 324:336 arg1 characterization 338:353 the comprehensive characterization 320:353 the comprehensive characterization of glycan structures 320:374 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 2 12 dep structure 256:264 arg1 the 252:254 the 252:254 the 252:254 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 3 13 theme High 410:413 arg1 technology 478:487 a well-established technology 459:487 a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans 459:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 13 theme High 410:413 arg1 HPLC 450:453 HPLC 450:453 HPLC 450:453 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 13 theme High 410:413 arg1 chromatography 434:447 High performance liquid chromatography 410:447 High performance liquid chromatography (HPLC) 410:454 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 2 14 theme analytical 390:399 arg1 methods 401:407 analytical methods 390:407 analytical methods 390:407 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 1 15 theme protein 95:101 arg1 glycosylation 103:115 protein glycosylation 95:115 protein glycosylation 95:115 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 3 16 theme O-linked 553:560 arg1 glycans 562:568 O-linked glycans 553:568 O-linked glycans 553:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 2 17 theme glycoproteins 297:309 arg1 relationship 281:292 functional relationship 270:292 functional relationship 270:292 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 2 17 theme glycoproteins 297:309 arg1 structure 256:264 structure 256:264 structure 256:264 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 3 18 theme structural 520:529 arg1 elucidation 531:541 the complete structural elucidation 507:541 the complete structural elucidation of N- and O-linked glycans 507:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 19 theme well-established 461:476 arg1 technology 478:487 a well-established technology 459:487 a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans 459:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 19 theme well-established 461:476 arg1 chromatography 434:447 High performance liquid chromatography 410:447 High performance liquid chromatography (HPLC) 410:454 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 4 20 with conjunction 770:780 arg1 initiative 802:811 the EUROCarbDB initiative 787:811 the EUROCarbDB initiative 787:811 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 3 21 theme major 606:610 arg1 analysis 584:591 the analysis 580:591 the analysis of data 580:599 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 21 theme major 606:610 arg1 bottleneck 612:621 a major bottleneck 604:621 a major bottleneck 604:621 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 1 22 theme indispensable 201:213 arg1 organism 191:198 an organism 188:198 an organism 188:198 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 0 23 theme HPLC-glycan 49:59 arg1 data 61:64 HPLC-glycan data 49:64 HPLC-glycan data 49:64 GlycoBase and autoGU: resources for interpreting HPLC-glycan data. 25753700 3 24 theme data 596:599 arg1 analysis 584:591 the analysis 580:591 the analysis of data 580:599 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 24 theme data 596:599 arg1 bottleneck 612:621 a major bottleneck 604:621 a major bottleneck 604:621 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 1 25 from indispensable 201:213 arg1 fields 232:237 many research fields 218:237 many research fields 218:237 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 2 26 theme structures 365:374 arg1 characterization 338:353 the comprehensive characterization 320:353 the comprehensive characterization of glycan structures 320:374 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 3 27 theme liquid 427:432 arg1 technology 478:487 a well-established technology 459:487 a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans 459:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 27 theme liquid 427:432 arg1 HPLC 450:453 HPLC 450:453 HPLC 450:453 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 27 theme liquid 427:432 arg1 chromatography 434:447 High performance liquid chromatography 410:447 High performance liquid chromatography (HPLC) 410:454 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 1 28 from fields 232:237 arg1 indispensable 201:213 indispensable 201:213 indispensable 201:213 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 4 29 theme HPLC-glycan 846:856 arg1 collections 863:873 HPLC-glycan data collections 846:873 HPLC-glycan data collections 846:873 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 25753700 2 30 theme glycan 358:363 arg1 structures 365:374 glycan structures 358:374 glycan structures 358:374 Determining the structure and functional relationship of glycoproteins requires the comprehensive characterization of glycan structures by a range of analytical methods. 25753700 3 31 theme robust 627:632 arg1 solutions 648:656 robust bioinformatic solutions 627:656 robust bioinformatic solutions 627:656 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 32 theme glycans 562:568 arg1 elucidation 531:541 the complete structural elucidation 507:541 the complete structural elucidation of N- and O-linked glycans 507:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 33 theme bioinformatic 634:646 arg1 solutions 648:656 robust bioinformatic solutions 627:656 robust bioinformatic solutions 627:656 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 1 34 from glycans 177:183 arg1 organism 191:198 an organism 188:198 an organism 188:198 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 1 35 theme many 218:221 arg1 fields 232:237 many research fields 218:237 many research fields 218:237 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 3 36 theme N- 546:547 arg1 elucidation 531:541 the complete structural elucidation 507:541 the complete structural elucidation of N- and O-linked glycans 507:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 1 37 theme research 223:230 arg1 fields 232:237 many research fields 218:237 many research fields 218:237 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 3 38 theme performance 415:425 arg1 technology 478:487 a well-established technology 459:487 a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans 459:568 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 38 theme performance 415:425 arg1 HPLC 450:453 HPLC 450:453 HPLC 450:453 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 3 38 theme performance 415:425 arg1 chromatography 434:447 High performance liquid chromatography 410:447 High performance liquid chromatography (HPLC) 410:454 High performance liquid chromatography (HPLC) is a well-established technology commonly used for the complete structural elucidation of N- and O-linked glycans; however, the analysis of data is a major bottleneck and robust bioinformatic solutions are required. 25753700 0 39 dep GlycoBase 0:8 arg1 resources 22:30 resources 22:30 GlycoBase and autoGU: resources for interpreting HPLC-glycan data. 0:65 GlycoBase and autoGU: resources for interpreting HPLC-glycan data. 25753700 1 40 theme comprehensive 141:153 arg1 study 155:159 the comprehensive study 137:159 the comprehensive study 137:159 The biological relevance of protein glycosylation has made glycomics, the comprehensive study to identify all glycans in an organism, indispensable in many research fields. 25753700 4 41 theme tools 729:733 arg1 availability 699:710 the availability 695:710 the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative 695:811 This chapter describes the availability of databases and tools, GlycoBase and autoGU developed in conjunction with the EUROCarbDB initiative, to assist the interpretation of HPLC-glycan data collections. 28397166 4 0 theme laser 886:890 arg1 desorption/ionization 892:912 multistage matrix-assisted laser desorption/ionization 859:912 multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn) 859:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 3 1 theme clinical 688:695 arg1 samples 697:703 clinical samples 688:703 clinical samples 688:703 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 3 2 theme qualitative 504:514 arg1 analysis 533:540 both qualitative and quantitative analysis 499:540 both qualitative and quantitative analysis 499:540 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 7 3 theme RA 1459:1460 arg1 diagnosis 1462:1470 RA diagnosis 1459:1470 RA diagnosis 1459:1470 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 1 4 from glycosylation 295:307 arg1 patients 338:345 rheumatoid arthritis (RA) patients 312:345 rheumatoid arthritis (RA) patients 312:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 4 5 theme glycans 845:851 arg1 identification 814:827 identification 814:827 identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE 814:985 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 6 theme multistage 859:868 arg1 desorption/ionization 892:912 multistage matrix-assisted laser desorption/ionization 859:912 multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn) 859:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 7 7 theme RA 1434:1435 arg1 studies 1447:1453 RA mechanism studies 1434:1453 RA mechanism studies 1434:1453 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 3 8 theme techniques 579:588 arg1 combination 554:564 a combination 552:564 a combination of different techniques 552:588 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 7 9 from glycosylation 1328:1340 arg1 patients 1355:1362 RA patients 1352:1362 RA patients compared with healthy people 1352:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 1 10 theme capillary 207:215 arg1 CE 234:235 CE 234:235 CE 234:235 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 10 theme capillary 207:215 arg1 electrophoresis 217:231 capillary electrophoresis 207:231 capillary electrophoresis (CE) 207:236 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 0 11 theme glycosylation 24:36 arg1 Characterization 0:15 Characterization 0:15 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 0:117 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 4 12 theme flight 922:927 arg1 MALDI-TOF-MSn 948:960 MALDI-TOF-MSn 948:960 MALDI-TOF-MSn 948:960 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 12 theme flight 922:927 arg1 spectrometry 934:945 flight mass spectrometry 922:945 flight mass spectrometry (MALDI-TOF-MSn) 922:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 13 theme present 713:719 arg1 study 721:725 the present study 709:725 the present study 709:725 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 8 14 gly glycosylation 1537:1549 arg1 patients 1575:1582 rheumatoid arthritis patients 1554:1582 rheumatoid arthritis patients 1554:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 4 15 theme spectrometry 934:945 arg1 quantitation 968:979 quantitation 968:979 quantitation by CE 968:985 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 15 theme spectrometry 934:945 arg1 time 914:917 multistage matrix-assisted laser desorption/ionization time 859:917 multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn) 859:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 1 16 gly glycosylation 295:307 arg1 patients 338:345 rheumatoid arthritis (RA) patients 312:345 rheumatoid arthritis (RA) patients 312:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 6 17 theme healthy 1242:1248 arg1 subjects 1250:1257 healthy subjects 1242:1257 healthy subjects 1242:1257 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 1 18 theme arthritis 323:331 arg1 patients 338:345 rheumatoid arthritis (RA) patients 312:345 rheumatoid arthritis (RA) patients 312:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 8 19 theme IgG 1533:1535 arg1 glycosylation 1537:1549 IgG glycosylation 1533:1549 IgG glycosylation in rheumatoid arthritis patients 1533:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 1 20 theme mass 179:182 arg1 MSn 198:200 MSn 198:200 MSn 198:200 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 20 theme mass 179:182 arg1 spectrometry 184:195 multistage mass spectrometry 168:195 multistage mass spectrometry (MSn) 168:201 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 0 21 from Characterization 0:15 arg1 patients 62:69 rheumatoid arthritis patients 41:69 rheumatoid arthritis patients 41:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 8 22 from analysis 1521:1528 arg1 patients 1575:1582 rheumatoid arthritis patients 1554:1582 rheumatoid arthritis patients 1554:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 8 23 theme arthritis 1565:1573 arg1 patients 1575:1582 rheumatoid arthritis patients 1554:1582 rheumatoid arthritis patients 1554:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 6 24 theme Further 1103:1109 arg1 quantitation 1120:1131 Further accurate quantitation 1103:1131 Further accurate quantitation 1103:1131 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 2 25 theme RA 428:429 arg1 immunopathogenesis 431:448 RA immunopathogenesis 428:448 RA immunopathogenesis 428:448 It has been recently suggested that IgG glycosylation defect may be involved in RA immunopathogenesis. 28397166 1 26 theme immunoglobulin 272:285 arg1 IgG 290:292 IgG 290:292 IgG 290:292 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 26 theme immunoglobulin 272:285 arg1 G 287:287 immunoglobulin G 272:287 immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients 272:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 3 27 theme Complete 451:458 arg1 characterization 460:475 Complete characterization 451:475 Complete characterization of glycans, including both qualitative and quantitative analysis, 451:541 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 0 28 theme IgG 20:22 arg1 glycosylation 24:36 IgG glycosylation 20:36 IgG glycosylation in rheumatoid arthritis patients 20:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 1 29 theme analytical 122:131 arg1 method 133:138 An analytical method 119:138 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) 119:236 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 7 30 theme RA 1352:1353 arg1 patients 1355:1362 RA patients 1352:1362 RA patients compared with healthy people 1352:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 2 31 theme glycosylation 388:400 arg1 defect 402:407 IgG glycosylation defect 384:407 IgG glycosylation defect 384:407 It has been recently suggested that IgG glycosylation defect may be involved in RA immunopathogenesis. 28397166 7 32 theme IgG 1345:1347 arg1 glycosylation 1328:1340 abnormal glycosylation 1319:1340 abnormal glycosylation of IgG in RA patients compared with healthy people 1319:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 3 33 theme high-throughput 628:642 arg1 methodologies 644:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 0 34 theme arthritis 52:60 arg1 patients 62:69 rheumatoid arthritis patients 41:69 rheumatoid arthritis patients 41:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 0 35 from glycosylation 24:36 arg1 patients 62:69 rheumatoid arthritis patients 41:69 rheumatoid arthritis patients 41:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 5 36 theme pattern 1057:1063 arg1 analysis 1065:1072 branching pattern analysis 1047:1072 branching pattern analysis 1047:1072 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 5 37 theme N-glycan 1010:1017 arg1 structures 1019:1028 the IgG N-glycan structures 1002:1028 the IgG N-glycan structures 1002:1028 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 4 38 theme matrix-assisted 870:884 arg1 desorption/ionization 892:912 multistage matrix-assisted laser desorption/ionization 859:912 multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn) 859:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 3 39 theme samples 697:703 arg1 analysis 676:683 analysis 676:683 analysis of clinical samples 676:703 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 4 40 theme desorption/ionization 892:912 arg1 time 914:917 multistage matrix-assisted laser desorption/ionization time 859:917 multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn) 859:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 6 41 theme IgG 1183:1185 arg1 N-glycans 1187:1195 IgG N-glycans 1183:1195 IgG N-glycans 1183:1195 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 1 42 theme spectrometry 184:195 arg1 CE 234:235 CE 234:235 CE 234:235 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 42 theme spectrometry 184:195 arg1 electrophoresis 217:231 capillary electrophoresis 207:231 capillary electrophoresis (CE) 207:236 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 42 theme spectrometry 184:195 arg1 combination 153:163 the combination 149:163 the combination of multistage mass spectrometry (MSn) 149:201 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 4 43 theme released 836:843 arg1 glycans 845:851 the released glycans 832:851 the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE 832:985 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 3 44 theme different 569:577 arg1 techniques 579:588 different techniques 569:588 different techniques 569:588 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 7 45 theme mechanism 1437:1445 arg1 studies 1447:1453 RA mechanism studies 1434:1453 RA mechanism studies 1434:1453 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 1 46 from analysis 260:267 arg1 patients 338:345 rheumatoid arthritis (RA) patients 312:345 rheumatoid arthritis (RA) patients 312:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 0 47 gly glycosylation 24:36 arg1 patients 62:69 rheumatoid arthritis patients 41:69 rheumatoid arthritis patients 41:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 6 48 theme RA 1200:1201 arg1 cases 1203:1207 RA cases 1200:1207 RA cases 1200:1207 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 6 49 theme N-glycans 1187:1195 arg1 sialylation 1168:1178 sialylation 1168:1178 sialylation 1168:1178 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 6 49 theme N-glycans 1187:1195 arg1 galactosylation 1148:1162 galactosylation 1148:1162 galactosylation 1148:1162 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 3 50 theme accurate 595:602 arg1 methodologies 644:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 8 51 theme Qualitative 1492:1502 arg1 analysis 1521:1528 Qualitative and quantitative analysis 1492:1528 Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients 1492:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 8 52 theme quantitative 1508:1519 arg1 analysis 1521:1528 Qualitative and quantitative analysis 1492:1528 Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients 1492:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 7 53 theme present 1407:1413 arg1 useful 1423:1428 useful 1423:1428 useful 1423:1428 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 7 53 theme present 1407:1413 arg1 work 1415:1418 the present work 1403:1418 the present work 1403:1418 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 3 54 theme sensitive 613:621 arg1 methodologies 644:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 4 55 theme mass 929:932 arg1 MALDI-TOF-MSn 948:960 MALDI-TOF-MSn 948:960 MALDI-TOF-MSn 948:960 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 55 theme mass 929:932 arg1 spectrometry 934:945 flight mass spectrometry 922:945 flight mass spectrometry (MALDI-TOF-MSn) 922:961 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 8 56 from glycosylation 1537:1549 arg1 patients 1575:1582 rheumatoid arthritis patients 1554:1582 rheumatoid arthritis patients 1554:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 3 57 theme robust 605:610 arg1 methodologies 644:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 accurate, robust, sensitive, and high-throughput methodologies 595:656 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 8 58 theme glycosylation 1537:1549 arg1 analysis 1521:1528 Qualitative and quantitative analysis 1492:1528 Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients 1492:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 0 59 theme capillary 92:100 arg1 electrophoresis 102:116 capillary electrophoresis 92:116 capillary electrophoresis 92:116 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 1 60 theme multistage 168:177 arg1 MSn 198:200 MSn 198:200 MSn 198:200 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 60 theme multistage 168:177 arg1 spectrometry 184:195 multistage mass spectrometry 168:195 multistage mass spectrometry (MSn) 168:201 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 8 61 theme rheumatoid 1554:1563 arg1 arthritis 1565:1573 rheumatoid arthritis 1554:1573 rheumatoid arthritis patients 1554:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 4 62 gly N-glycosylation 728:742 arg1 IgG 747:749 IgG 747:749 IgG 747:749 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 62 gly N-glycosylation 728:742 arg1 patients 757:764 RA patients 754:764 RA patients 754:764 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 62 gly N-glycosylation 728:742 arg1 study 721:725 the present study 709:725 the present study 709:725 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 62 gly N-glycosylation 728:742 arg1 people 781:786 healthy people 773:786 healthy people 773:786 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 0 63 from patients 62:69 arg1 Characterization 0:15 Characterization 0:15 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 0:117 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 4 64 theme RA 754:755 arg1 patients 757:764 RA patients 754:764 RA patients 754:764 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 5 65 with MSn 1077:1079 arg1 high-throughput 1086:1100 high-throughput 1086:1100 high-throughput 1086:1100 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 6 66 theme accurate 1111:1118 arg1 quantitation 1120:1131 Further accurate quantitation 1103:1131 Further accurate quantitation 1103:1131 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 8 67 theme capillary 1605:1613 arg1 electrophoresis 1615:1629 capillary electrophoresis 1605:1629 capillary electrophoresis 1605:1629 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 6 68 gly sialylation 1168:1178 arg1 N-glycans 1187:1195 IgG N-glycans 1183:1195 IgG N-glycans 1183:1195 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 6 68 gly sialylation 1168:1178 arg1 cases 1203:1207 RA cases 1200:1207 RA cases 1200:1207 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 1 69 from patients 338:345 arg1 analysis 260:267 the analysis 256:267 the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients 256:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 70 theme G 287:287 arg1 glycosylation 295:307 immunoglobulin G (IgG) glycosylation 272:307 immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients 272:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 4 71 from N-glycosylation 728:742 arg1 patients 757:764 RA patients 754:764 RA patients 754:764 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 71 from N-glycosylation 728:742 arg1 people 781:786 healthy people 773:786 healthy people 773:786 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 4 72 theme IgG 747:749 arg1 N-glycosylation 728:742 N-glycosylation 728:742 N-glycosylation of IgG in RA patients and in healthy people 728:786 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 0 73 theme rheumatoid 41:50 arg1 patients 62:69 rheumatoid arthritis patients 41:69 rheumatoid arthritis patients 41:69 Characterization of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 6 74 from sialylation 1168:1178 arg1 cases 1203:1207 RA cases 1200:1207 RA cases 1200:1207 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 2 75 theme IgG 384:386 arg1 defect 402:407 IgG glycosylation defect 384:407 IgG glycosylation defect 384:407 It has been recently suggested that IgG glycosylation defect may be involved in RA immunopathogenesis. 28397166 8 76 from patients 1575:1582 arg1 analysis 1521:1528 Qualitative and quantitative analysis 1492:1528 Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients 1492:1582 Graphical Abstract Qualitative and quantitative analysis of IgG glycosylation in rheumatoid arthritis patients by MALDI-TOF-MSn and capillary electrophoresis. 28397166 1 77 theme glycosylation 295:307 arg1 analysis 260:267 the analysis 256:267 the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients 256:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 7 78 gly glycosylation 1328:1340 arg1 IgG 1345:1347 IgG 1345:1347 IgG 1345:1347 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 7 78 gly glycosylation 1328:1340 arg1 patients 1355:1362 RA patients 1352:1362 RA patients compared with healthy people 1352:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 5 79 theme branching 1047:1055 arg1 analysis 1065:1072 branching pattern analysis 1047:1072 branching pattern analysis 1047:1072 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 7 80 theme abnormal 1319:1326 arg1 glycosylation 1328:1340 abnormal glycosylation 1319:1340 abnormal glycosylation of IgG in RA patients compared with healthy people 1319:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 1 81 theme rheumatoid 312:321 arg1 RA 334:335 RA 334:335 RA 334:335 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 1 81 theme rheumatoid 312:321 arg1 arthritis 323:331 rheumatoid arthritis 312:331 rheumatoid arthritis (RA) patients 312:345 An analytical method based on the combination of multistage mass spectrometry (MSn) and capillary electrophoresis (CE) was developed for the analysis of immunoglobulin G (IgG) glycosylation in rheumatoid arthritis (RA) patients. 28397166 5 82 theme IgG 1006:1008 arg1 structures 1019:1028 the IgG N-glycan structures 1002:1028 the IgG N-glycan structures 1002:1028 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 4 83 theme healthy 773:779 arg1 people 781:786 healthy people 773:786 healthy people 773:786 In the present study, N-glycosylation of IgG in RA patients and in healthy people was characterized through identification of the released glycans using multistage matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MSn), and quantitation by CE. 28397166 3 84 theme glycans 480:486 arg1 characterization 460:475 Complete characterization 451:475 Complete characterization of glycans, including both qualitative and quantitative analysis, 451:541 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 28397166 5 85 theme structures 1019:1028 arg1 Assignment 988:997 Assignment 988:997 Assignment of the IgG N-glycan structures 988:1028 Assignment of the IgG N-glycan structures was made through branching pattern analysis by MSn with high-throughput. 28397166 7 86 theme healthy 1378:1384 arg1 people 1386:1391 healthy people 1378:1391 healthy people 1378:1391 The results indicate that CE coupled with MSn can identify abnormal glycosylation of IgG in RA patients compared with healthy people, and that the present work is useful for RA mechanism studies and RA diagnosis. 28397166 6 87 from galactosylation 1148:1162 arg1 cases 1203:1207 RA cases 1200:1207 RA cases 1200:1207 Further accurate quantitation indicated that galactosylation and sialylation of IgG N-glycans in RA cases were significantly lower than in healthy subjects. 28397166 3 88 theme quantitative 520:531 arg1 analysis 533:540 both qualitative and quantitative analysis 499:540 both qualitative and quantitative analysis 499:540 Complete characterization of glycans, including both qualitative and quantitative analysis, requires a combination of different techniques, and accurate, robust, sensitive, and high-throughput methodologies are important for analysis of clinical samples. 26983412 3 0 from improvements 553:564 arg1 humanization 573:584 the humanization 569:584 the humanization of the N-glycans in a recently developed S. cerevisiae strain 569:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 1 1 theme relevant 184:191 arg1 proteins 193:200 therapeutic and other industrially relevant proteins 149:200 therapeutic and other industrially relevant proteins 149:200 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 3 2 theme S. 627:628 arg1 strain 641:646 a recently developed S. cerevisiae strain 606:646 a recently developed S. cerevisiae strain 606:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 4 3 theme N-acetylglucosamine 835:853 arg1 transferases 855:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 6 4 theme glycoengineered 1069:1083 arg1 strain 1085:1090 the glycoengineered strain 1065:1090 the glycoengineered strain 1065:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 7 5 from N-glycosylation 1422:1436 arg1 cerevisiae 1444:1453 S. cerevisiae 1441:1453 S. cerevisiae 1441:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 4 6 theme trimannosyl 676:686 arg1 oligosaccharide 701:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 7 7 theme target 1181:1186 arg1 glycan 1188:1193 the complex-type target glycan 1164:1193 the complex-type target glycan 1164:1193 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 0 8 from structure 67:75 arg1 cerevisiae 94:103 Saccharomyces cerevisiae 80:103 Saccharomyces cerevisiae 80:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 6 9 attach present 1054:1060 arg1 strain 1085:1090 the glycoengineered strain 1065:1090 the glycoengineered strain 1065:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 6 9 attach present 1054:1060 arg2 structures 1043:1052 the interfering structures 1027:1052 the interfering structures present in the glycoengineered strain 1027:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 1 10 gly glycoprotein 315:326 arg1 glycoprotein 315:326 alternative, non-mammalian glycoprotein expression systems 288:345 alternative, non-mammalian glycoprotein expression systems 288:345 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 4 11 theme glycan 784:789 arg1 formation 791:799 complex-type glycan formation 771:799 complex-type glycan formation 771:799 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 6 12 theme MNN1 1127:1130 arg1 gene 1132:1135 the MNN1 gene 1123:1135 the MNN1 gene 1123:1135 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 0 13 theme human-type 47:56 arg1 structure 67:75 a human-type N-glycan structure 45:75 a human-type N-glycan structure in Saccharomyces cerevisiae 45:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 5 14 theme efficiency 977:986 arg1 terms 938:942 terms 938:942 terms of glycoform homogeneity and the efficiency of complex-type glycosylation 938:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 0 15 theme structure 67:75 arg1 homogeneity 30:40 homogeneity 30:40 homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae 30:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 1 16 theme other 165:169 arg1 proteins 193:200 therapeutic and other industrially relevant proteins 149:200 therapeutic and other industrially relevant proteins 149:200 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 8 17 theme MNN1 1480:1483 arg1 deletion 1485:1492 the MNN1 deletion 1476:1492 the MNN1 deletion 1476:1492 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 0 18 theme Saccharomyces 80:92 arg1 cerevisiae 94:103 Saccharomyces cerevisiae 80:103 Saccharomyces cerevisiae 80:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 2 19 theme protein 445:451 arg1 production 453:462 therapeutic protein production 433:462 therapeutic protein production 433:462 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 0 20 theme N-glycan 58:65 arg1 structure 67:75 a human-type N-glycan structure 45:75 a human-type N-glycan structure in Saccharomyces cerevisiae 45:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 4 21 theme apparatus-targeted 810:827 arg1 transferases 855:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 3 22 from strain 641:646 arg1 humanization 573:584 the humanization 569:584 the humanization of the N-glycans in a recently developed S. cerevisiae strain 569:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 5 23 theme glycoengineered 907:921 arg1 strain 923:928 the glycoengineered strain 903:928 the glycoengineered strain 903:928 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 1 24 theme expression 328:337 arg1 systems 339:345 alternative, non-mammalian glycoprotein expression systems 288:345 alternative, non-mammalian glycoprotein expression systems 288:345 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 6 25 from strain 1085:1090 arg1 present 1054:1060 present 1054:1060 present 1054:1060 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 8 26 theme transporter 1542:1552 arg1 expression 1502:1511 the expression 1498:1511 the expression of a UDP-N-acetylglucosamine transporter 1498:1552 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 8 26 theme transporter 1542:1552 arg1 combination 1461:1471 a combination 1459:1471 a combination of the MNN1 deletion 1459:1492 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 7 27 from expression 1216:1225 arg1 lactis 1287:1292 Kluyveromyces lactis 1273:1292 Kluyveromyces lactis 1273:1292 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 28 theme transporter 1256:1266 arg1 expression 1216:1225 the expression 1212:1225 the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis 1212:1292 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 6 29 from present 1054:1060 arg1 strain 1085:1090 the glycoengineered strain 1065:1090 the glycoengineered strain 1065:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 1 30 theme N-glycosylation 226:240 arg1 pathway 242:248 the N-glycosylation pathway 222:248 the N-glycosylation pathway 222:248 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 0 31 gly homogeneity 30:40 arg1 structure 67:75 a human-type N-glycan structure 45:75 a human-type N-glycan structure in Saccharomyces cerevisiae 45:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 7 32 theme efficient 1399:1407 arg1 N-glycosylation 1422:1436 efficient complex-type N-glycosylation 1399:1436 efficient complex-type N-glycosylation in S. cerevisiae 1399:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 5 33 dep strain 923:928 arg1 both 930:933 both 930:933 both 930:933 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 5 34 theme glycoform 947:955 arg1 homogeneity 957:967 glycoform homogeneity 947:967 glycoform homogeneity 947:967 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 2 35 theme established 399:409 arg1 cerevisiae 376:385 Saccharomyces cerevisiae 362:385 Saccharomyces cerevisiae 362:385 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 2 35 theme established 399:409 arg1 organism 416:423 the most established host organism 390:423 the most established host organism used in therapeutic protein production 390:462 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 3 36 theme N-glycans 593:601 arg1 humanization 573:584 the humanization 569:584 the humanization of the N-glycans in a recently developed S. cerevisiae strain 569:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 1 37 theme alternative 288:298 arg1 systems 339:345 alternative, non-mammalian glycoprotein expression systems 288:345 alternative, non-mammalian glycoprotein expression systems 288:345 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 5 38 gly homogeneity 957:967 arg1 glycosylation 1004:1016 complex-type glycosylation 991:1016 complex-type glycosylation 991:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 3 39 from humanization 573:584 arg1 strain 641:646 a recently developed S. cerevisiae strain 606:646 a recently developed S. cerevisiae strain 606:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 1 40 theme proteins 193:200 arg1 feature 138:144 an important feature 125:144 an important feature of therapeutic and other industrially relevant proteins 125:200 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 1 40 theme proteins 193:200 arg1 N-glycosylation 106:120 N-glycosylation 106:120 N-glycosylation 106:120 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 5 41 theme glycan 885:890 arg1 pattern 892:898 the glycan pattern 881:898 the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation 881:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 7 42 theme glycan 1188:1193 arg1 abundance 1151:1159 The relative abundance 1138:1159 The relative abundance of the complex-type target glycan 1138:1193 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 3 43 theme developed 617:625 arg1 strain 641:646 a recently developed S. cerevisiae strain 606:646 a recently developed S. cerevisiae strain 606:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 5 44 theme homogeneity 957:967 arg1 terms 938:942 terms 938:942 terms of glycoform homogeneity and the efficiency of complex-type glycosylation 938:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 7 45 theme complex-type 1168:1179 arg1 glycan 1188:1193 the complex-type target glycan 1164:1193 the complex-type target glycan 1164:1193 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 5 46 theme strain 923:928 arg1 pattern 892:898 the glycan pattern 881:898 the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation 881:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 5 47 theme complex-type 991:1002 arg1 glycosylation 1004:1016 complex-type glycosylation 991:1016 complex-type glycosylation 991:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 4 48 theme tailored 667:674 arg1 oligosaccharide 701:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 6 49 theme present 1054:1060 arg1 structures 1043:1052 the interfering structures 1027:1052 the interfering structures present in the glycoengineered strain 1027:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 8 50 theme improved 1614:1621 arg1 homogeneity 1623:1633 a significantly improved homogeneity 1598:1633 a significantly improved homogeneity 1598:1633 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 4 51 theme lipid-linked 688:699 arg1 oligosaccharide 701:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 7 52 theme relative 1142:1149 arg1 abundance 1151:1159 The relative abundance 1138:1159 The relative abundance of the complex-type target glycan 1138:1193 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 3 53 theme cerevisiae 630:639 arg1 strain 641:646 a recently developed S. cerevisiae strain 606:646 a recently developed S. cerevisiae strain 606:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 5 54 theme glycosylation 1004:1016 arg1 homogeneity 957:967 glycoform homogeneity 947:967 glycoform homogeneity 947:967 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 5 54 theme glycosylation 1004:1016 arg1 efficiency 977:986 the efficiency 973:986 the efficiency of complex-type glycosylation 973:1016 We improved the glycan pattern of the glycoengineered strain both in terms of glycoform homogeneity and the efficiency of complex-type glycosylation. 26983412 6 55 theme interfering 1031:1041 arg1 structures 1043:1052 the interfering structures 1027:1052 the interfering structures present in the glycoengineered strain 1027:1090 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 1 56 dep alternative 288:298 arg1 non-mammalian 301:313 non-mammalian 301:313 non-mammalian 301:313 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 1 57 theme important 128:136 arg1 feature 138:144 an important feature 125:144 an important feature of therapeutic and other industrially relevant proteins 125:200 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 1 57 theme important 128:136 arg1 N-glycosylation 106:120 N-glycosylation 106:120 N-glycosylation 106:120 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 6 58 theme gene 1132:1135 arg1 deletion 1111:1118 deletion 1111:1118 deletion of the MNN1 gene 1111:1135 Most of the interfering structures present in the glycoengineered strain were eliminated by deletion of the MNN1 gene. 26983412 0 59 from cerevisiae 94:103 arg1 homogeneity 30:40 homogeneity 30:40 homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae 30:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 4 60 link lipid-linked 688:699 arg1 oligosaccharide 701:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 a tailored trimannosyl lipid-linked oligosaccharide 665:715 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 4 61 theme complex-type 771:782 arg1 formation 791:799 complex-type glycan formation 771:799 complex-type glycan formation 771:799 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 7 62 theme Golgi 1358:1362 arg1 apparatus 1364:1372 the Golgi apparatus 1354:1372 the Golgi apparatus 1354:1372 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 9 63 theme further 1673:1679 arg1 step 1681:1684 a further step 1671:1684 a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae 1671:1767 Our results represent a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae. 26983412 9 64 from homogeneity 1740:1750 arg1 cerevisiae 1758:1767 S. cerevisiae 1755:1767 S. cerevisiae 1755:1767 Our results represent a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae. 26983412 7 65 theme UDP-N-acetylglucosamine 1325:1347 arg1 factor 1388:1393 a limiting factor 1377:1393 a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae 1377:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 65 theme UDP-N-acetylglucosamine 1325:1347 arg1 import 1315:1320 the import 1311:1320 the import of UDP-N-acetylglucosamine into the Golgi apparatus 1311:1372 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 3 66 from N-glycans 593:601 arg1 strain 641:646 a recently developed S. cerevisiae strain 606:646 a recently developed S. cerevisiae strain 606:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 9 67 theme humanized 1704:1712 arg1 glycoproteins 1714:1726 humanized glycoproteins 1704:1726 humanized glycoproteins 1704:1726 Our results represent a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae. 26983412 7 68 from lactis 1287:1292 arg1 expression 1216:1225 the expression 1212:1225 the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis 1212:1292 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 68 from lactis 1287:1292 arg1 transporter 1256:1266 a UDP-N-acetylglucosamine transporter 1230:1266 a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis 1230:1292 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 4 69 theme human 829:833 arg1 transferases 855:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 0 70 from homogeneity 30:40 arg1 cerevisiae 94:103 Saccharomyces cerevisiae 80:103 Saccharomyces cerevisiae 80:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 8 71 theme deletion 1485:1492 arg1 expression 1502:1511 the expression 1498:1511 the expression of a UDP-N-acetylglucosamine transporter 1498:1552 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 8 71 theme deletion 1485:1492 arg1 combination 1461:1471 a combination 1459:1471 a combination of the MNN1 deletion 1459:1492 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 2 72 theme therapeutic 433:443 arg1 production 453:462 therapeutic protein production 433:462 therapeutic protein production 433:462 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 0 73 theme dual 2:5 arg1 approach 7:14 A dual approach 0:14 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae 0:103 A dual approach for improving homogeneity of a human-type N-glycan structure in Saccharomyces cerevisiae. 26983412 9 74 theme high 1735:1738 arg1 homogeneity 1740:1750 a high homogeneity 1733:1750 a high homogeneity in S. cerevisiae 1733:1767 Our results represent a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae. 26983412 9 75 gly glycoproteins 1714:1726 arg1 glycoproteins 1714:1726 humanized glycoproteins 1704:1726 humanized glycoproteins 1704:1726 Our results represent a further step towards obtaining humanized glycoproteins with a high homogeneity in S. cerevisiae. 26983412 1 76 theme glycoprotein 315:326 arg1 systems 339:345 alternative, non-mammalian glycoprotein expression systems 288:345 alternative, non-mammalian glycoprotein expression systems 288:345 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 2 77 theme interesting 481:491 arg1 host 493:496 an interesting host 478:496 an interesting host for glycoengineering 478:517 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 4 78 theme Golgi 804:808 arg1 transferases 855:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 Golgi apparatus-targeted human N-acetylglucosamine transferases 804:866 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 8 79 theme UDP-N-acetylglucosamine 1518:1540 arg1 transporter 1542:1552 a UDP-N-acetylglucosamine transporter 1516:1552 a UDP-N-acetylglucosamine transporter 1516:1552 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 7 80 theme UDP-N-acetylglucosamine 1232:1254 arg1 transporter 1256:1266 a UDP-N-acetylglucosamine transporter 1230:1266 a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis 1230:1292 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 81 gly N-glycosylation 1422:1436 arg1 cerevisiae 1444:1453 S. cerevisiae 1441:1453 S. cerevisiae 1441:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 82 theme complex-type 1409:1420 arg1 N-glycosylation 1422:1436 efficient complex-type N-glycosylation 1399:1436 efficient complex-type N-glycosylation in S. cerevisiae 1399:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 1 83 theme therapeutic 149:159 arg1 proteins 193:200 therapeutic and other industrially relevant proteins 149:200 therapeutic and other industrially relevant proteins 149:200 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 3 84 theme further 545:551 arg1 improvements 553:564 further improvements 545:564 further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain 545:646 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 4 85 dep transferred 731:741 arg1 followed 759:766 followed 759:766 followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases 759:866 In this strain, a tailored trimannosyl lipid-linked oligosaccharide is formed and transferred to the protein, followed by complex-type glycan formation by Golgi apparatus-targeted human N-acetylglucosamine transferases. 26983412 1 86 theme pathway 242:248 arg1 engineering 207:217 engineering 207:217 engineering of the N-glycosylation pathway 207:248 N-glycosylation is an important feature of therapeutic and other industrially relevant proteins, and engineering of the N-glycosylation pathway provides opportunities for developing alternative, non-mammalian glycoprotein expression systems. 26983412 3 87 attach present 537:543 arg2 we 534:535 we 534:535 we 534:535 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 3 87 attach present 537:543 arg1 work 528:531 this work 523:531 this work 523:531 In this work, we present further improvements in the humanization of the N-glycans in a recently developed S. cerevisiae strain. 26983412 7 88 theme limiting 1379:1386 arg1 factor 1388:1393 a limiting factor 1377:1393 a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae 1377:1453 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 7 88 theme limiting 1379:1386 arg1 import 1315:1320 the import 1311:1320 the import of UDP-N-acetylglucosamine into the Golgi apparatus 1311:1372 The relative abundance of the complex-type target glycan was increased by the expression of a UDP-N-acetylglucosamine transporter from Kluyveromyces lactis, indicating that the import of UDP-N-acetylglucosamine into the Golgi apparatus is a limiting factor for efficient complex-type N-glycosylation in S. cerevisiae. 26983412 8 89 theme complex-type 1572:1583 arg1 glycans 1585:1591 complex-type glycans 1572:1591 complex-type glycans 1572:1591 By a combination of the MNN1 deletion and the expression of a UDP-N-acetylglucosamine transporter, a strain forming complex-type glycans with a significantly improved homogeneity was obtained. 26983412 2 90 theme host 411:414 arg1 cerevisiae 376:385 Saccharomyces cerevisiae 362:385 Saccharomyces cerevisiae 362:385 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26983412 2 90 theme host 411:414 arg1 organism 416:423 the most established host organism 390:423 the most established host organism used in therapeutic protein production 390:462 Among yeasts, Saccharomyces cerevisiae is the most established host organism used in therapeutic protein production and therefore an interesting host for glycoengineering. 26576925 0 0 theme glycoform 77:85 arg1 ratio 87:91 glycoform ratio 77:91 glycoform ratio 77:91 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 2 1 contain has 258:260 arg1 PrP 251:253 PrP 251:253 PrP(C) 251:256 PrP(C) has two sialylated N-linked carbohydrates. 26576925 2 1 contain has 258:260 arg2 carbohydrates 286:298 two sialylated N-linked carbohydrates 262:298 two sialylated N-linked carbohydrates 262:298 PrP(C) has two sialylated N-linked carbohydrates. 26576925 2 1 contain has 258:260 arg1 C 255:255 C 255:255 C 255:255 PrP(C) has two sialylated N-linked carbohydrates. 26576925 8 2 gly desialylation 1264:1276 arg1 C 1285:1285 C 1285:1285 C 1285:1285 Moreover, as predicted by the hypothesis, partial desialylation of PrP(C) significantly increased the replication rate. 26576925 8 2 gly desialylation 1264:1276 arg1 PrP 1281:1283 PrP 1281:1283 PrP(C) 1281:1286 Moreover, as predicted by the hypothesis, partial desialylation of PrP(C) significantly increased the replication rate. 26576925 1 3 theme proteinaceous 116:128 arg1 agent 141:145 a proteinaceous infectious agent 114:145 a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 114:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 3 theme proteinaceous 116:128 arg1 Prion 94:98 Prion or PrP 94:105 Prion 94:98 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 3 theme proteinaceous 116:128 arg1 PrP 103:105 Prion or PrP 94:105 PrP 103:105 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 4 theme called 219:224 arg1 protein 232:238 a sialoglycoprotein called prion protein 199:238 a sialoglycoprotein called prion protein 199:238 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 3 5 from charge 406:411 arg1 surface 420:426 the surface 416:426 the surface of prion particles 416:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 1 6 theme infectious 130:139 arg1 agent 141:145 a proteinaceous infectious agent 114:145 a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 114:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 6 theme infectious 130:139 arg1 Prion 94:98 Prion or PrP 94:105 Prion 94:98 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 6 theme infectious 130:139 arg1 PrP 103:105 Prion or PrP 94:105 PrP 103:105 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 1 7 theme prion 226:230 arg1 protein 232:238 a sialoglycoprotein called prion protein 199:238 a sialoglycoprotein called prion protein 199:238 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 6 8 gly desialylated 858:869 arg1 C 875:875 C 875:875 C 875:875 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 8 gly desialylated 858:869 arg1 PrP 871:873 partially desialylated PrP 848:873 partially desialylated PrP(C) 848:876 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 8 gly desialylated 858:869 arg1 substrate 892:900 a substrate 890:900 a substrate 890:900 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 9 9 gly sialylation 1362:1372 arg1 glycans 1386:1392 N-linked glycans 1377:1392 N-linked glycans 1377:1392 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 9 10 theme glycoform 1465:1473 arg1 ratios 1475:1480 glycoform ratios 1465:1480 glycoform ratios 1465:1480 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 3 11 theme particles 437:445 arg1 surface 420:426 the surface 416:426 the surface of prion particles 416:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 9 12 theme replication 1444:1454 arg1 rate 1456:1459 replication rate 1444:1459 replication rate 1444:1459 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 1 13 theme protein 232:238 arg1 form 191:194 a misfolded and aggregated form 164:194 a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 164:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 7 14 theme hypersialylated 1040:1054 arg1 molecules 1056:1064 hypersialylated molecules 1040:1064 hypersialylated molecules 1040:1064 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 1 15 gly sialoglycoprotein 201:217 arg1 sialoglycoprotein 201:217 sialoglycoprotein 201:217 sialoglycoprotein 201:217 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 9 16 theme N-linked 1377:1384 arg1 glycans 1386:1392 N-linked glycans 1377:1392 N-linked glycans 1377:1392 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 2 17 link N-linked 277:284 arg1 carbohydrates 286:298 two sialylated N-linked carbohydrates 262:298 two sialylated N-linked carbohydrates 262:298 PrP(C) has two sialylated N-linked carbohydrates. 26576925 7 18 with strains 1108:1114 arg1 constraints 1146:1156 the strongest structural constraints 1121:1156 the strongest structural constraints 1121:1156 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 7 18 with strains 1108:1114 arg1 strategy 1161:1168 a strategy 1159:1168 a strategy that helped reduce electrostatic repulsion 1159:1211 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 3 19 theme acid 372:375 arg1 residues 377:384 the terminal sialic acid residues 352:384 the terminal sialic acid residues creating a negative charge on the surface of prion particles 352:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 9 20 theme prion 1404:1408 arg1 barrier 1422:1428 a prion replication barrier 1402:1428 a prion replication barrier that controls replication rate and glycoform ratios and has broad implications 1402:1507 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 4 21 theme new 477:479 arg1 hypothesis 481:490 a new hypothesis 475:490 a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio 475:633 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 3 22 theme prion 431:435 arg1 particles 437:445 prion particles 431:445 prion particles 431:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 4 23 theme structural 553:562 arg1 constraints 564:574 structural constraints 553:574 structural constraints that control prion replication and PrP(Sc) glycoform ratio 553:633 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 7 24 theme structural 1135:1144 arg1 constraints 1146:1156 the strongest structural constraints 1121:1156 the strongest structural constraints 1121:1156 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 7 24 theme structural 1135:1144 arg1 strategy 1161:1168 a strategy 1159:1168 a strategy that helped reduce electrostatic repulsion 1159:1211 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 4 25 theme current 452:458 arg1 study 460:464 The current study 448:464 The current study 448:464 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 5 26 gly monoglycosylated 764:779 arg1 C 785:785 C 785:785 C 785:785 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 5 26 gly monoglycosylated 764:779 arg1 PrP 781:783 monoglycosylated PrP 764:783 monoglycosylated PrP(C) 764:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 6 27 theme glycoforms 924:933 arg1 recruitment 903:913 recruitment 903:913 recruitment of three glycoforms into PrP(Sc) 903:946 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 1 28 theme PrP 243:245 arg1 form 191:194 a misfolded and aggregated form 164:194 a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 164:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 5 29 contain have 722:725 arg2 groups 739:744 more sialic groups 727:744 more sialic groups 727:744 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 5 29 contain have 722:725 arg1 molecules 707:715 diglycosylated PrP(C) molecules 685:715 diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) 685:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 6 30 theme respective 986:995 arg1 populations 997:1007 their respective populations 980:1007 their respective populations in the substrate 980:1024 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 4 31 theme prion 589:593 arg1 replication 595:605 prion replication 589:605 prion replication 589:605 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 0 32 theme prion 19:23 arg1 glycans 33:39 the prion protein glycans 15:39 the prion protein glycans 15:39 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 9 33 theme broad 1490:1494 arg1 implications 1496:1507 broad implications 1490:1507 broad implications 1490:1507 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 9 34 link N-linked 1377:1384 arg1 glycans 1386:1392 N-linked glycans 1377:1392 N-linked glycans 1377:1392 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 5 35 theme diglycosylated 685:698 arg1 C 704:704 C 704:704 C 704:704 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 5 35 theme diglycosylated 685:698 arg1 PrP 700:702 diglycosylated PrP 685:702 diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) 685:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 3 36 theme sialic 365:370 arg1 residues 377:384 the terminal sialic acid residues 352:384 the terminal sialic acid residues creating a negative charge on the surface of prion particles 352:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 2 37 theme N-linked 277:284 arg1 carbohydrates 286:298 two sialylated N-linked carbohydrates 262:298 two sialylated N-linked carbohydrates 262:298 PrP(C) has two sialylated N-linked carbohydrates. 26576925 6 38 from populations 997:1007 arg1 substrate 1016:1024 the substrate 1012:1024 the substrate 1012:1024 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 1 39 theme misfolded 166:174 arg1 form 191:194 a misfolded and aggregated form 164:194 a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 164:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 0 40 theme glycans 33:39 arg1 Sialylation 0:10 Sialylation 0:10 Sialylation of the prion protein glycans 0:39 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 7 41 theme strongest 1125:1133 arg1 constraints 1146:1156 the strongest structural constraints 1121:1156 the strongest structural constraints 1121:1156 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 7 41 theme strongest 1125:1133 arg1 strategy 1161:1168 a strategy 1159:1168 a strategy that helped reduce electrostatic repulsion 1159:1211 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 4 42 dep hypothesis 481:490 arg1 creates 545:551 creates 545:551 creates structural constraints that control prion replication and PrP(Sc) glycoform ratio 545:633 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 3 43 dep directed 329:336 arg1 outward 338:344 outward 338:344 outward 338:344 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 5 44 theme more 727:730 arg1 groups 739:744 more sialic groups 727:744 more sialic groups 727:744 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 0 45 theme protein 25:31 arg1 glycans 33:39 the prion protein glycans 15:39 the prion protein glycans 15:39 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 5 46 theme sialic 732:737 arg1 groups 739:744 more sialic groups 727:744 more sialic groups 727:744 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 1 47 theme aggregated 180:189 arg1 form 191:194 a misfolded and aggregated form 164:194 a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C) 164:248 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 0 48 theme prion 50:54 arg1 rate 68:71 prion replication rate 50:71 prion replication rate 50:71 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 5 49 theme monoglycosylated 764:779 arg1 C 785:785 C 785:785 C 785:785 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 5 49 theme monoglycosylated 764:779 arg1 PrP 781:783 monoglycosylated PrP 764:783 monoglycosylated PrP(C) 764:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 8 50 theme partial 1256:1262 arg1 desialylation 1264:1276 partial desialylation 1256:1276 partial desialylation of PrP(C) 1256:1286 Moreover, as predicted by the hypothesis, partial desialylation of PrP(C) significantly increased the replication rate. 26576925 2 51 theme sialylated 266:275 arg1 carbohydrates 286:298 two sialylated N-linked carbohydrates 262:298 two sialylated N-linked carbohydrates 262:298 PrP(C) has two sialylated N-linked carbohydrates. 26576925 5 52 theme PrP 700:702 arg1 molecules 707:715 diglycosylated PrP(C) molecules 685:715 diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) 685:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 9 53 theme glycans 1386:1392 arg1 sialylation 1362:1372 sialylation 1362:1372 sialylation of N-linked glycans 1362:1392 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 2 54 gly sialylated 266:275 arg1 carbohydrates 286:298 two sialylated N-linked carbohydrates 262:298 two sialylated N-linked carbohydrates 262:298 PrP(C) has two sialylated N-linked carbohydrates. 26576925 8 55 theme replication 1316:1326 arg1 rate 1328:1331 the replication rate 1312:1331 the replication rate 1312:1331 Moreover, as predicted by the hypothesis, partial desialylation of PrP(C) significantly increased the replication rate. 26576925 6 56 used used 882:885 arg2 PrP 871:873 partially desialylated PrP 848:873 partially desialylated PrP(C) 848:876 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 56 used used 882:885 arg2 substrate 892:900 a substrate 890:900 a substrate 890:900 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 56 used used 882:885 arg2 C 875:875 C 875:875 C 875:875 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 3 57 theme negative 397:404 arg1 charge 406:411 a negative charge 395:411 a negative charge on the surface of prion particles 395:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 8 58 theme PrP 1281:1283 arg1 desialylation 1264:1276 partial desialylation 1256:1276 partial desialylation of PrP(C) 1256:1286 Moreover, as predicted by the hypothesis, partial desialylation of PrP(C) significantly increased the replication rate. 26576925 4 59 theme glycoform 619:627 arg1 ratio 629:633 PrP(Sc) glycoform ratio 611:633 PrP(Sc) glycoform ratio 611:633 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 9 60 contain has 1486:1488 arg2 implications 1496:1507 broad implications 1490:1507 broad implications 1490:1507 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 9 60 contain has 1486:1488 arg1 barrier 1422:1428 a prion replication barrier 1402:1428 a prion replication barrier that controls replication rate and glycoform ratios and has broad implications 1402:1507 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 9 61 theme replication 1410:1420 arg1 barrier 1422:1428 a prion replication barrier 1402:1428 a prion replication barrier that controls replication rate and glycoform ratios and has broad implications 1402:1507 This study illustrates that sialylation of N-linked glycans creates a prion replication barrier that controls replication rate and glycoform ratios and has broad implications. 26576925 1 62 dep called 219:224 arg1 sialoglycoprotein 201:217 sialoglycoprotein 201:217 sialoglycoprotein 201:217 Prion or PrP(Sc) is a proteinaceous infectious agent that consists of a misfolded and aggregated form of a sialoglycoprotein called prion protein or PrP(C). 26576925 7 63 gly hypersialylated 1040:1054 arg1 molecules 1056:1064 hypersialylated molecules 1040:1064 hypersialylated molecules 1040:1064 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 4 64 theme electrostatic 497:509 arg1 repulsion 511:519 electrostatic repulsion 497:519 electrostatic repulsion between sialic residues 497:543 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 5 65 theme hypothesis 655:664 arg1 support 639:645 support 639:645 support of this hypothesis 639:664 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 3 66 theme terminal 356:363 arg1 residues 377:384 the terminal sialic acid residues 352:384 the terminal sialic acid residues creating a negative charge on the surface of prion particles 352:445 In PrP(Sc), the glycans are directed outward, with the terminal sialic acid residues creating a negative charge on the surface of prion particles. 26576925 7 67 theme electrostatic 1189:1201 arg1 repulsion 1203:1211 electrostatic repulsion 1189:1211 electrostatic repulsion 1189:1211 In addition, hypersialylated molecules were also excluded from conversion in the strains with the strongest structural constraints, a strategy that helped reduce electrostatic repulsion. 26576925 4 68 theme PrP 611:613 arg1 ratio 629:633 PrP(Sc) glycoform ratio 611:633 PrP(Sc) glycoform ratio 611:633 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 4 69 theme sialic 529:534 arg1 residues 536:543 sialic residues 529:543 sialic residues 529:543 The current study proposes a new hypothesis that electrostatic repulsion between sialic residues creates structural constraints that control prion replication and PrP(Sc) glycoform ratio. 26576925 0 70 theme replication 56:66 arg1 rate 68:71 prion replication rate 50:71 prion replication rate 50:71 Sialylation of the prion protein glycans controls prion replication rate and glycoform ratio. 26576925 5 71 gly diglycosylated 685:698 arg1 C 704:704 C 704:704 C 704:704 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 5 71 gly diglycosylated 685:698 arg1 PrP 700:702 diglycosylated PrP 685:702 diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) 685:786 In support of this hypothesis, here we show that diglycosylated PrP(C) molecules that have more sialic groups per molecule than monoglycosylated PrP(C) were preferentially excluded from conversion. 26576925 6 72 theme desialylated 858:869 arg1 C 875:875 C 875:875 C 875:875 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 72 theme desialylated 858:869 arg1 PrP 871:873 partially desialylated PrP 848:873 partially desialylated PrP(C) 848:876 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 26576925 6 72 theme desialylated 858:869 arg1 substrate 892:900 a substrate 890:900 a substrate 890:900 However, when partially desialylated PrP(C) was used as a substrate, recruitment of three glycoforms into PrP(Sc) was found to be proportional to their respective populations in the substrate. 28994289 1 0 theme chromatography 320:333 arg1 techniques 242:251 the combined techniques 229:251 the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography 229:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 1 1 theme rapa 217:220 arg1 L. 222:223 the common edible plant Brassica rapa L. 184:223 the common edible plant Brassica rapa L. 184:223 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 5 2 dep proliferation 912:924 arg1 TNF-α 972:976 TNF-α 972:976 TNF-α 972:976 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 2 dep proliferation 912:924 arg1 IL-6 963:966 IL-6 963:966 IL-6 963:966 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 0 3 from Characterization 11:26 arg1 L 97:97 Brassica rapa L 83:97 Brassica rapa L 83:97 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 2 4 theme composition 351:361 arg1 analysis 363:370 Monosaccharide composition analysis 336:370 Monosaccharide composition analysis 336:370 Monosaccharide composition analysis showed that BRNP-1 and BRNP-2 were composed of glucosyl residues. 28994289 3 5 theme α-D- 563:566 arg1 residues 587:594 α-D-(1 → 4)-linked Glcp residues 563:594 α-D-(1 → 4)-linked Glcp residues 563:594 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 5 6 theme cytokine 943:950 arg1 secretion 952:960 cytokine secretion 943:960 cytokine secretion 943:960 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 1 7 dep BRNP-1 131:136 arg1 kDa 160:162 4.8 kDa 156:162 4.8 kDa 156:162 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 1 7 dep BRNP-1 131:136 arg1 kDa 143:145 6.9 kDa 139:145 6.9 kDa 139:145 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 1 8 dep kDa 143:145 arg1 BRNP-2 148:153 BRNP-2 148:153 BRNP-2 148:153 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 5 9 theme NO 927:928 arg1 release 930:936 NO release 927:936 NO release 927:936 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 3 10 attach attached 666:673 arg2 residues 657:664 terminally linked Glcp residues 634:664 terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 634:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 10 attach attached 666:673 arg3 position 686:693 the O-6 position 678:693 the O-6 position of backbone-glycosyl residues 678:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 11 theme backbone-glycosyl 698:714 arg1 residues 716:723 backbone-glycosyl residues 698:723 backbone-glycosyl residues 698:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 1 12 dep polysaccharides 114:128 arg1 BRNP-1 131:136 BRNP-1 131:136 BRNP-1 131:136 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 3 13 link -linked 574:580 arg1 residues 587:594 α-D-(1 → 4)-linked Glcp residues 563:594 α-D-(1 → 4)-linked Glcp residues 563:594 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 14 theme O-6 682:684 arg1 position 686:693 the O-6 position 678:693 the O-6 position of backbone-glycosyl residues 678:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 6 15 theme macrophage-mediated 1063:1081 arg1 responses 1090:1098 macrophage-mediated immune responses 1063:1098 macrophage-mediated immune responses 1063:1098 These results suggested that BRNP-1 and BRNP-2 may enhance macrophage-mediated immune responses. 28994289 0 16 dep   99:99 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization 0:26 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 0 16 dep   99:99 arg1 Activity 50:57 Immunostimulatory Activity 32:57 Immunostimulatory Activity 32:57 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 3 17 theme 1D- 454:456 arg1 analyses 469:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 0 18 from Activity 50:57 arg1 L 97:97 Brassica rapa L 83:97 Brassica rapa L 83:97 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 5 19 theme BRNP-2 890:895 arg1 dosage 857:862 the higher dosage 846:862 the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 846:895 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 19 theme BRNP-2 890:895 arg1 μg/mL 869:873 400 μg/mL 865:873 400 μg/mL 865:873 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 0 20 theme Structural 0:9 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization 0:26 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 1 21 theme combined 233:240 arg1 techniques 242:251 the combined techniques 229:251 the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography 229:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 3 22 theme Methylation 438:448 arg1 analyses 469:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 0 23 theme Immunostimulatory 32:48 arg1 Activity 50:57 Immunostimulatory Activity 32:57 Immunostimulatory Activity 32:57 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 5 24 theme Bioassay 802:809 arg1 results 811:817 Bioassay results 802:817 Bioassay results 802:817 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 3 25 theme residues 716:723 arg1 position 686:693 the O-6 position 678:693 the O-6 position of backbone-glycosyl residues 678:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 1 26 theme neutral 106:112 arg1 polysaccharides 114:128 Two neutral polysaccharides 102:128 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) 102:163 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 2 27 theme glucosyl 419:426 arg1 residues 428:435 glucosyl residues 419:435 glucosyl residues 419:435 Monosaccharide composition analysis showed that BRNP-1 and BRNP-2 were composed of glucosyl residues. 28994289 1 28 theme ion-exchange 256:267 arg1 chromatography 269:282 ion-exchange chromatography 256:282 ion-exchange chromatography 256:282 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 0 29 from L 97:97 arg1 Polysaccharides 62:76 Polysaccharides 62:76 Polysaccharides from Brassica rapa L 62:97 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 0 29 from L 97:97 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization 0:26 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 0 29 from L 97:97 arg1 Activity 50:57 Immunostimulatory Activity 32:57 Immunostimulatory Activity 32:57 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 4 30 theme molecular 784:792 arg1 weight 794:799 molecular weight 784:799 molecular weight 784:799 BRNP-1 and BRNP-2, however, differed in branch degree and molecular weight. 28994289 5 31 theme higher 850:855 arg1 dosage 857:862 the higher dosage 846:862 the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 846:895 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 31 theme higher 850:855 arg1 μg/mL 869:873 400 μg/mL 865:873 400 μg/mL 865:873 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 1 32 theme chromatography 269:282 arg1 techniques 242:251 the combined techniques 229:251 the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography 229:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 3 33 theme -linked 574:580 arg1 residues 587:594 α-D-(1 → 4)-linked Glcp residues 563:594 α-D-(1 → 4)-linked Glcp residues 563:594 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 34 theme 2D-NMR 462:467 arg1 analyses 469:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses 438:476 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 2 35 theme Monosaccharide 336:349 arg1 analysis 363:370 Monosaccharide composition analysis 336:370 Monosaccharide composition analysis 336:370 Monosaccharide composition analysis showed that BRNP-1 and BRNP-2 were composed of glucosyl residues. 28994289 0 36 theme Polysaccharides 62:76 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization 0:26 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 0 36 theme Polysaccharides 62:76 arg1 Activity 50:57 Immunostimulatory Activity 32:57 Immunostimulatory Activity 32:57 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 1 37 theme high-performance 288:303 arg1 chromatography 320:333 high-performance gel permeation chromatography 288:333 high-performance gel permeation chromatography 288:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 3 38 contain contained 515:523 arg1 BRNP-1 497:502 BRNP-1 497:502 BRNP-1 497:502 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 38 contain contained 515:523 arg2 chain 536:540 a backbone chain 525:540 a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 525:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 38 contain contained 515:523 arg1 BRNP-2 508:513 BRNP-2 508:513 BRNP-2 508:513 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 39 theme linked 645:650 arg1 residues 657:664 terminally linked Glcp residues 634:664 terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 634:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 5 40 theme RAW264.7 982:989 arg1 macrophages 991:1001 RAW264.7 macrophages 982:1001 RAW264.7 macrophages 982:1001 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 6 41 theme immune 1083:1088 arg1 responses 1090:1098 macrophage-mediated immune responses 1063:1098 macrophage-mediated immune responses 1063:1098 These results suggested that BRNP-1 and BRNP-2 may enhance macrophage-mediated immune responses. 28994289 1 42 theme gel 305:307 arg1 chromatography 320:333 high-performance gel permeation chromatography 288:333 high-performance gel permeation chromatography 288:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 5 43 with treatment 831:839 arg1 dosage 857:862 the higher dosage 846:862 the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 846:895 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 43 with treatment 831:839 arg1 μg/mL 869:873 400 μg/mL 865:873 400 μg/mL 865:873 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 3 44 theme Glcp 652:655 arg1 residues 657:664 terminally linked Glcp residues 634:664 terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 634:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 5 45 theme BRNP-1 879:884 arg1 dosage 857:862 the higher dosage 846:862 the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 846:895 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 45 theme BRNP-1 879:884 arg1 μg/mL 869:873 400 μg/mL 865:873 400 μg/mL 865:873 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 1 46 theme common 188:193 arg1 L. 222:223 the common edible plant Brassica rapa L. 184:223 the common edible plant Brassica rapa L. 184:223 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 5 47 theme macrophages 991:1001 arg1 secretion 952:960 cytokine secretion 943:960 cytokine secretion 943:960 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 47 theme macrophages 991:1001 arg1 proliferation 912:924 proliferation 912:924 proliferation 912:924 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 5 47 theme macrophages 991:1001 arg1 release 930:936 NO release 927:936 NO release 927:936 Bioassay results showed that treatment with the higher dosage (400 μg/mL) of BRNP-1 and BRNP-2 stimulated the proliferation, NO release, and cytokine secretion (IL-6 and TNF-α) of RAW264.7 macrophages. 28994289 3 48 theme side 600:603 arg1 chains 605:610 side chains 600:610 side chains 600:610 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 49 dep α-D- 563:566 arg1 4 572:572 4 572:572 4 572:572 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 49 dep α-D- 563:566 arg1 1 568:568 1 568:568 1 568:568 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 3 50 link linked 645:650 arg1 residues 657:664 terminally linked Glcp residues 634:664 terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 634:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 1 51 theme edible 195:200 arg1 L. 222:223 the common edible plant Brassica rapa L. 184:223 the common edible plant Brassica rapa L. 184:223 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 0 52 theme rapa 92:95 arg1 L 97:97 Brassica rapa L 83:97 Brassica rapa L 83:97 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 3 53 theme Glcp 582:585 arg1 residues 587:594 α-D-(1 → 4)-linked Glcp residues 563:594 α-D-(1 → 4)-linked Glcp residues 563:594 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 1 54 theme plant 202:206 arg1 L. 222:223 the common edible plant Brassica rapa L. 184:223 the common edible plant Brassica rapa L. 184:223 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 0 55 theme Brassica 83:90 arg1 L 97:97 Brassica rapa L 83:97 Brassica rapa L 83:97 Structural Characterization and Immunostimulatory Activity of Polysaccharides from Brassica rapa L. . 28994289 1 56 theme permeation 309:318 arg1 chromatography 320:333 high-performance gel permeation chromatography 288:333 high-performance gel permeation chromatography 288:333 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 28994289 4 57 theme branch 766:771 arg1 degree 773:778 branch degree 766:778 branch degree 766:778 BRNP-1 and BRNP-2, however, differed in branch degree and molecular weight. 28994289 3 58 theme backbone 527:534 arg1 chain 536:540 a backbone chain 525:540 a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues 525:723 Methylation and 1D- and 2D-NMR analyses revealed that both BRNP-1 and BRNP-2 contained a backbone chain that was composed of α-D-(1 → 4)-linked Glcp residues and side chains that were composed of terminally linked Glcp residues attached at the O-6 position of backbone-glycosyl residues. 28994289 1 59 theme Brassica 208:215 arg1 L. 222:223 the common edible plant Brassica rapa L. 184:223 the common edible plant Brassica rapa L. 184:223 Two neutral polysaccharides (BRNP-1, 6.9 kDa; BRNP-2, 4.8 kDa) were purified from the common edible plant Brassica rapa L. via the combined techniques of ion-exchange chromatography and high-performance gel permeation chromatography. 24308717 6 0 theme N-glycan 1052:1059 arg1 pool 1061:1064 the N-glycan pool 1048:1064 the N-glycan pool 1048:1064 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 4 1 theme N-glycan 653:660 arg1 pool 662:665 The N-glycan pool 649:665 The N-glycan pool 649:665 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 1 2 gly glycosylated 205:216 arg1 protein 240:246 a highly glycosylated therapeutic Fc-fusion protein 196:246 a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites 196:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 2 gly glycosylated 205:216 arg1 Etanercept 182:191 Etanercept 182:191 Etanercept 182:191 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 10 3 theme Electron-transfer 1485:1501 arg1 dissociation 1503:1514 Electron-transfer dissociation 1485:1514 Electron-transfer dissociation (ETD) 1485:1520 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 10 3 theme Electron-transfer 1485:1501 arg1 ETD 1517:1519 ETD 1517:1519 ETD 1517:1519 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 9 4 from residues 1375:1382 arg1 O-glycans 1391:1399 the O-glycans 1387:1399 the O-glycans 1387:1399 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 8 5 attach attached 1289:1296 arg1 residues 1309:1316 Ser/Thr residues 1301:1316 Ser/Thr residues 1301:1316 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 8 5 attach attached 1289:1296 arg2 These 1218:1222 These 1218:1222 These 1218:1222 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 2 6 gly glycosylation 337:349 arg1 etanercept 354:363 etanercept 354:363 etanercept 354:363 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 11 7 theme biotherapeutics 1815:1829 arg1 production 1780:1789 the production 1776:1789 the production of safe and efficacious biotherapeutics 1776:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 3 8 theme N-glycosidase 607:619 arg1 F 621:621 peptide N-glycosidase F 599:621 peptide N-glycosidase F 599:621 Etanercept was first treated with peptide N-glycosidase F to release the N-glycans. 24308717 1 9 theme therapeutic 218:228 arg1 protein 240:246 a highly glycosylated therapeutic Fc-fusion protein 196:246 a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites 196:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 9 theme therapeutic 218:228 arg1 Etanercept 182:191 Etanercept 182:191 Etanercept 182:191 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 6 10 theme array 1016:1020 arg1 digestions 1022:1031 exoglycosidase array digestions 1001:1031 exoglycosidase array digestions of aliquots of the N-glycan pool 1001:1064 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 9 11 theme sialic 1363:1368 arg1 residues 1375:1382 the sialic acid residues 1359:1382 the sialic acid residues on the O-glycans 1359:1399 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 4 12 theme 2-AB 704:707 arg1 tag 723:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 0 13 theme electron-transfer 136:152 arg1 functionality 167:179 electron-transfer dissociation functionality 136:179 electron-transfer dissociation functionality 136:179 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 10 14 used used 1531:1534 arg2 ETD 1517:1519 ETD 1517:1519 ETD 1517:1519 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 10 14 used used 1531:1534 arg2 dissociation 1503:1514 Electron-transfer dissociation 1485:1514 Electron-transfer dissociation (ETD) 1485:1520 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 4 15 theme fluorescence 710:721 arg1 tag 723:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 1 16 theme N- 271:272 arg1 sites 294:298 multiple N- and O-glycosylation sites 262:298 multiple N- and O-glycosylation sites 262:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 11 17 theme future 1686:1691 arg1 studies 1693:1699 future studies 1686:1699 future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics 1686:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 7 18 from removal 1071:1077 arg1 etanercept 1101:1110 etanercept 1101:1110 etanercept 1101:1110 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 11 19 theme O-glycosylation 1629:1643 arg1 determination 1591:1603 The determination 1587:1603 The determination of N- and O-glycans and O-glycosylation sites in etanercept 1587:1663 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 7 20 theme selective 1128:1136 arg1 characterization 1138:1153 the selective characterization 1124:1153 the selective characterization of O-glycopeptides 1124:1172 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 10 21 gly O-glycosylation 1564:1578 arg2 12 1552:1553 12 1552:1553 12 1552:1553 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 10 21 gly O-glycosylation 1564:1578 arg2 sites 1580:1584 the 12 occupied O-glycosylation sites 1548:1584 the 12 occupied O-glycosylation sites 1548:1584 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 4 22 theme liquid 764:769 arg1 chromatography 817:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography 747:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC) 747:843 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 4 22 theme liquid 764:769 arg1 UPLC-HILIC 833:842 UPLC-HILIC 833:842 UPLC-HILIC 833:842 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 11 23 theme O-glycans 1615:1623 arg1 determination 1591:1603 The determination 1587:1603 The determination of N- and O-glycans and O-glycosylation sites in etanercept 1587:1663 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 11 24 dep N- 1608:1609 arg1 sites 1645:1649 sites 1645:1649 sites 1645:1649 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 4 25 theme interaction 798:808 arg1 chromatography 817:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography 747:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC) 747:843 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 4 25 theme interaction 798:808 arg1 UPLC-HILIC 833:842 UPLC-HILIC 833:842 UPLC-HILIC 833:842 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 0 26 theme quadrupole 78:87 arg1 spectrometry 109:120 quadrupole time-of-flight mass spectrometry 78:120 quadrupole time-of-flight mass spectrometry 78:120 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 9 27 theme LC/MS 1434:1438 arg1 protocol 1443:1450 an automated LC/MS(E) protocol 1421:1450 an automated LC/MS(E) protocol 1421:1450 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 1 28 gly O-glycosylation 278:292 arg2 sites 294:298 multiple N- and O-glycosylation sites 262:298 multiple N- and O-glycosylation sites 262:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 10 29 theme O-glycosylation 1564:1578 arg1 sites 1580:1584 the 12 occupied O-glycosylation sites 1548:1584 the 12 occupied O-glycosylation sites 1548:1584 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 0 30 theme mass 104:107 arg1 spectrometry 109:120 quadrupole time-of-flight mass spectrometry 78:120 quadrupole time-of-flight mass spectrometry 78:120 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 9 31 theme E 1440:1440 arg1 protocol 1443:1450 an automated LC/MS(E) protocol 1421:1450 an automated LC/MS(E) protocol 1421:1450 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 2 32 link O-linked 500:507 arg1 glycans 509:515 O-linked glycans 500:515 O-linked glycans 500:515 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 9 33 theme protocol 1443:1450 arg1 use 1414:1416 the use 1410:1416 the use of an automated LC/MS(E) protocol to identify the O-glycopeptides 1410:1482 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 2 34 dep N- 493:494 arg1 sites 558:562 the occupied O-glycosylation sites 529:562 the occupied O-glycosylation sites 529:562 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 2 34 dep N- 493:494 arg1 the 489:491 the 489:491 the 489:491 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 2 35 theme systematic 448:457 arg1 approach 459:466 a systematic approach 446:466 a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites 446:562 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 0 36 theme N- 0:1 arg1 analysis 23:30 N- and O-glycosylation analysis 0:30 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry 0:120 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 11 37 theme N- 1608:1609 arg1 determination 1591:1603 The determination 1587:1603 The determination of N- and O-glycans and O-glycosylation sites in etanercept 1587:1663 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 0 38 theme O-glycosylation 7:21 arg1 analysis 23:30 N- and O-glycosylation analysis 0:30 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry 0:120 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 3 39 theme peptide 599:605 arg1 F 621:621 peptide N-glycosidase F 599:621 peptide N-glycosidase F 599:621 Etanercept was first treated with peptide N-glycosidase F to release the N-glycans. 24308717 6 40 theme exoglycosidase 1001:1014 arg1 digestions 1022:1031 exoglycosidase array digestions 1001:1031 exoglycosidase array digestions of aliquots of the N-glycan pool 1001:1064 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 2 41 theme occupied 533:540 arg1 sites 558:562 the occupied O-glycosylation sites 529:562 the occupied O-glycosylation sites 529:562 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 11 42 from determination 1591:1603 arg1 etanercept 1654:1663 etanercept 1654:1663 etanercept 1654:1663 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 11 43 theme protein 1750:1756 arg1 glycosylations 1758:1771 specific protein glycosylations 1741:1771 specific protein glycosylations 1741:1771 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 2 44 theme chromatography/mass 394:412 arg1 spectrometry 414:425 liquid chromatography/mass spectrometry 387:425 liquid chromatography/mass spectrometry (LC/MS) methods 387:441 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 2 44 theme chromatography/mass 394:412 arg1 LC/MS 428:432 LC/MS 428:432 LC/MS 428:432 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 6 45 theme linkage 962:968 arg1 information 970:980 linkage information 962:980 linkage information 962:980 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 6 46 theme pool 1061:1064 arg1 aliquots 1036:1043 aliquots 1036:1043 aliquots of the N-glycan pool 1036:1064 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 11 47 theme efficacious 1803:1813 arg1 biotherapeutics 1815:1829 safe and efficacious biotherapeutics 1794:1829 safe and efficacious biotherapeutics 1794:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 9 48 theme α2→3,6,8,9 1319:1328 arg1 sialidase 1330:1338 α2→3,6,8,9 sialidase 1319:1338 α2→3,6,8,9 sialidase 1319:1338 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 11 49 theme safe 1794:1797 arg1 biotherapeutics 1815:1829 safe and efficacious biotherapeutics 1794:1829 safe and efficacious biotherapeutics 1794:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 6 50 theme monosaccharide 934:947 arg1 sequence 949:956 monosaccharide sequence 934:956 monosaccharide sequence 934:956 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 2 51 theme etanercept 354:363 arg1 glycosylation 337:349 the glycosylation 333:349 the glycosylation of etanercept 333:363 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 8 52 theme Ser/Thr 1301:1307 arg1 residues 1309:1316 Ser/Thr residues 1301:1316 Ser/Thr residues 1301:1316 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 6 53 theme aliquots 1036:1043 arg1 digestions 1022:1031 exoglycosidase array digestions 1001:1031 exoglycosidase array digestions of aliquots of the N-glycan pool 1001:1064 These assignments, which included monosaccharide sequence and linkage information, were confirmed by exoglycosidase array digestions of aliquots of the N-glycan pool. 24308717 2 54 theme glycosylation 337:349 arg1 characterization 313:328 An in-depth characterization 301:328 An in-depth characterization of the glycosylation of etanercept 301:363 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 1 55 theme glycosylated 205:216 arg1 protein 240:246 a highly glycosylated therapeutic Fc-fusion protein 196:246 a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites 196:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 55 theme glycosylated 205:216 arg1 Etanercept 182:191 Etanercept 182:191 Etanercept 182:191 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 11 56 theme biological 1716:1725 arg1 importance 1727:1736 the biological importance 1712:1736 the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics 1712:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 1 57 theme Fc-fusion 230:238 arg1 protein 240:246 a highly glycosylated therapeutic Fc-fusion protein 196:246 a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites 196:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 57 theme Fc-fusion 230:238 arg1 Etanercept 182:191 Etanercept 182:191 Etanercept 182:191 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 0 58 theme dissociation 154:165 arg1 functionality 167:179 electron-transfer dissociation functionality 136:179 electron-transfer dissociation functionality 136:179 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 9 59 theme acid 1370:1373 arg1 residues 1375:1382 the sialic acid residues 1359:1382 the sialic acid residues on the O-glycans 1359:1399 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 11 60 from importance 1727:1736 arg1 production 1780:1789 the production 1776:1789 the production of safe and efficacious biotherapeutics 1776:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 7 61 from etanercept 1101:1110 arg1 removal 1071:1077 The removal 1067:1077 The removal of the N-glycans from etanercept 1067:1110 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 7 61 from etanercept 1101:1110 arg1 N-glycans 1086:1094 the N-glycans 1082:1094 the N-glycans from etanercept 1082:1110 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 1 62 theme multiple 262:269 arg1 sites 294:298 multiple N- and O-glycosylation sites 262:298 multiple N- and O-glycosylation sites 262:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 4 63 theme liquid 810:815 arg1 chromatography 817:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography 747:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC) 747:843 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 4 63 theme liquid 810:815 arg1 UPLC-HILIC 833:842 UPLC-HILIC 833:842 UPLC-HILIC 833:842 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 7 64 theme O-glycopeptides 1158:1172 arg1 characterization 1138:1153 the selective characterization 1124:1153 the selective characterization of O-glycopeptides 1124:1172 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 2 65 theme in-depth 304:311 arg1 characterization 313:328 An in-depth characterization 301:328 An in-depth characterization of the glycosylation of etanercept 301:363 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 4 66 theme ultraperformance 747:762 arg1 chromatography 817:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography 747:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC) 747:843 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 4 66 theme ultraperformance 747:762 arg1 UPLC-HILIC 833:842 UPLC-HILIC 833:842 UPLC-HILIC 833:842 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 9 67 theme automated 1424:1432 arg1 protocol 1443:1450 an automated LC/MS(E) protocol 1421:1450 an automated LC/MS(E) protocol 1421:1450 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 2 68 theme O-glycosylation 542:556 arg1 sites 558:562 the occupied O-glycosylation sites 529:562 the occupied O-glycosylation sites 529:562 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 4 69 theme chromatography-hydrophilic 771:796 arg1 chromatography 817:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography 747:830 ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC) 747:843 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 4 69 theme chromatography-hydrophilic 771:796 arg1 UPLC-HILIC 833:842 UPLC-HILIC 833:842 UPLC-HILIC 833:842 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 10 70 theme occupied 1555:1562 arg1 sites 1580:1584 the 12 occupied O-glycosylation sites 1548:1584 the 12 occupied O-glycosylation sites 1548:1584 Electron-transfer dissociation (ETD) was then used to pinpoint the 12 occupied O-glycosylation sites. 24308717 5 71 theme Preliminary 846:856 arg1 structures 858:867 Preliminary structures 846:867 Preliminary structures 846:867 Preliminary structures were assigned using Glycobase. 24308717 7 72 theme N-glycans 1086:1094 arg1 removal 1071:1077 The removal 1067:1077 The removal of the N-glycans from etanercept 1067:1110 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 9 73 gly O-glycopeptides 1468:1482 arg2 O-glycopeptides 1468:1482 the O-glycopeptides 1464:1482 the O-glycopeptides 1464:1482 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 2 74 theme O-linked 500:507 arg1 glycans 509:515 O-linked glycans 500:515 O-linked glycans 500:515 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 9 75 used used 1344:1347 arg2 sialidase 1330:1338 α2→3,6,8,9 sialidase 1319:1338 α2→3,6,8,9 sialidase 1319:1338 α2→3,6,8,9 sialidase was used to remove the sialic acid residues on the O-glycans allowing the use of an automated LC/MS(E) protocol to identify the O-glycopeptides. 24308717 2 76 gly O-glycosylation 542:556 arg2 sites 558:562 the occupied O-glycosylation sites 529:562 the occupied O-glycosylation sites 529:562 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 0 77 theme etanercept 35:44 arg1 analysis 23:30 N- and O-glycosylation analysis 0:30 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry 0:120 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 8 78 theme core 1250:1253 arg1 subtype 1257:1263 the core 1 subtype 1246:1263 the core 1 subtype (HexHexNAc O-structure) 1246:1287 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 8 78 theme core 1250:1253 arg1 O-structure 1276:1286 HexHexNAc O-structure 1266:1286 HexHexNAc O-structure 1266:1286 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 11 79 theme specific 1741:1748 arg1 glycosylations 1758:1771 specific protein glycosylations 1741:1771 specific protein glycosylations 1741:1771 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 24308717 0 80 theme liquid 52:57 arg1 chromatography 59:72 liquid chromatography 52:72 liquid chromatography 52:72 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 0 81 gly O-glycosylation 7:21 arg1 etanercept 35:44 etanercept 35:44 etanercept 35:44 N- and O-glycosylation analysis of etanercept using liquid chromatography and quadrupole time-of-flight mass spectrometry equipped with electron-transfer dissociation functionality. 24308717 7 82 gly O-glycopeptides 1158:1172 arg2 O-glycopeptides 1158:1172 O-glycopeptides 1158:1172 O-glycopeptides 1158:1172 The removal of the N-glycans from etanercept facilitated the selective characterization of O-glycopeptides and enabled the O-glycans to be identified. 24308717 2 83 theme spectrometry 414:425 arg1 methods 435:441 liquid chromatography/mass spectrometry (LC/MS) methods 387:441 liquid chromatography/mass spectrometry (LC/MS) methods 387:441 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 8 84 theme HexHexNAc 1266:1274 arg1 subtype 1257:1263 the core 1 subtype 1246:1263 the core 1 subtype (HexHexNAc O-structure) 1246:1287 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 8 84 theme HexHexNAc 1266:1274 arg1 O-structure 1276:1286 HexHexNAc O-structure 1266:1286 HexHexNAc O-structure 1266:1286 These were predominantly of the core 1 subtype (HexHexNAc O-structure) attached to Ser/Thr residues. 24308717 1 85 contain contains 253:260 arg1 protein 240:246 a highly glycosylated therapeutic Fc-fusion protein 196:246 a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites 196:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 85 contain contains 253:260 arg1 Etanercept 182:191 Etanercept 182:191 Etanercept 182:191 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 1 85 contain contains 253:260 arg2 sites 294:298 multiple N- and O-glycosylation sites 262:298 multiple N- and O-glycosylation sites 262:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 2 86 theme liquid 387:392 arg1 spectrometry 414:425 liquid chromatography/mass spectrometry 387:425 liquid chromatography/mass spectrometry (LC/MS) methods 387:441 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 2 86 theme liquid 387:392 arg1 LC/MS 428:432 LC/MS 428:432 LC/MS 428:432 An in-depth characterization of the glycosylation of etanercept was carried out using liquid chromatography/mass spectrometry (LC/MS) methods in a systematic approach in which we analyzed the N- and O-linked glycans and located the occupied O-glycosylation sites. 24308717 1 87 theme O-glycosylation 278:292 arg1 sites 294:298 multiple N- and O-glycosylation sites 262:298 multiple N- and O-glycosylation sites 262:298 Etanercept is a highly glycosylated therapeutic Fc-fusion protein that contains multiple N- and O-glycosylation sites. 24308717 4 88 theme 2-aminobenzamide 686:701 arg1 tag 723:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 a 2-aminobenzamide (2-AB) fluorescence tag 684:725 The N-glycan pool was labeled with a 2-aminobenzamide (2-AB) fluorescence tag and separated using ultraperformance liquid chromatography-hydrophilic interaction liquid chromatography (UPLC-HILIC). 24308717 11 89 theme glycosylations 1758:1771 arg1 importance 1727:1736 the biological importance 1712:1736 the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics 1712:1829 The determination of N- and O-glycans and O-glycosylation sites in etanercept provides a basis for future studies addressing the biological importance of specific protein glycosylations in the production of safe and efficacious biotherapeutics. 25484058 3 0 theme 2013 549:552 arg1 end 542:544 the end 538:544 the end of 2013 538:552 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 1 1 theme monoclonal 160:169 arg1 mAb 181:183 mAb 181:183 mAb 181:183 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 1 1 theme monoclonal 160:169 arg1 antibody 171:178 monoclonal antibody 160:178 monoclonal antibody (mAb) therapeutics 160:197 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 2 2 theme drugs 308:312 arg1 drugs 308:312 the 10 top-selling drugs 289:312 the 10 top-selling drugs 289:312 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 2 2 theme drugs 308:312 arg1 Six 282:284 Six 282:284 Six 282:284 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 2 2 theme drugs 308:312 arg1 therapeutics 333:344 antibody-based therapeutics 318:344 antibody-based therapeutics that will lose patent protection soon 318:382 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 7 3 theme single 1253:1258 arg1 injection 1267:1275 only a single sample injection 1246:1275 only a single sample injection of 200 fmol 1246:1287 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 7 4 theme amino 1319:1323 arg1 AA 1332:1333 AA 1332:1333 AA 1332:1333 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 7 4 theme amino 1319:1323 arg1 acids 1325:1329 100% amino acids 1314:1329 100% amino acids (AA) sequence characterization 1314:1360 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 5 5 theme reference 960:968 arg1 mAbs 970:973 marketed reference mAbs 951:973 marketed reference mAbs 951:973 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 4 6 with glycoproteins 573:585 arg1 range 599:603 a wide range 592:603 a wide range of micro-variants 592:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 2 7 theme antibody-based 318:331 arg1 drugs 308:312 the 10 top-selling drugs 289:312 the 10 top-selling drugs 289:312 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 2 7 theme antibody-based 318:331 arg1 therapeutics 333:344 antibody-based therapeutics 318:344 antibody-based therapeutics that will lose patent protection soon 318:382 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 2 7 theme antibody-based 318:331 arg1 Six 282:284 Six 282:284 Six 282:284 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 11 8 theme single 2146:2151 arg1 injection 2153:2161 a single injection 2144:2161 a single injection of each candidate enabling improvements in the biosimilar development pipeline 2144:2240 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 4 9 theme structure 704:712 arg1 assessment 714:723 structure assessment 704:723 structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming 704:821 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 11 10 theme development 2221:2231 arg1 pipeline 2233:2240 the biosimilar development pipeline 2206:2240 the biosimilar development pipeline 2206:2240 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 1 11 theme strong 238:243 arg1 potency 257:263 their strong therapeutic potency 232:263 their strong therapeutic potency 232:263 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 11 12 from improvements 2190:2201 arg1 pipeline 2233:2240 the biosimilar development pipeline 2206:2240 the biosimilar development pipeline 2206:2240 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 1 13 theme antibody 171:178 arg1 therapeutics 186:197 monoclonal antibody (mAb) therapeutics 160:197 monoclonal antibody (mAb) therapeutics 160:197 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 6 14 theme biosimilar 1144:1153 arg1 candidates 1155:1164 their biosimilar candidates 1138:1164 their biosimilar candidates 1138:1164 CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. 25484058 4 15 theme biosimilarity 761:773 arg1 studies 775:781 manufacturing control and biosimilarity studies 735:781 studies 775:781 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 0 16 theme zone 90:93 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 4 17 gly glycoproteins 573:585 arg1 glycoproteins 573:585 highly complex glycoproteins 558:585 highly complex glycoproteins with a wide range of micro-variants 558:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 4 17 gly glycoproteins 573:585 arg1 characterization 647:662 extensive characterization 637:662 extensive characterization 637:662 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 5 18 theme respective 985:994 arg1 candidate 1007:1015 their respective biosimilar candidate 979:1015 their respective biosimilar candidate 979:1015 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 8 19 theme methodology 1664:1674 arg1 capacity 1638:1645 the capacity 1634:1645 the capacity of the developed methodology to detect up to 16 different glycans 1634:1711 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 7 20 dep enabled 1306:1312 arg1 allows 1369:1374 allows 1369:1374 allows a difference of even one AA between 2 samples to be distinguished precisely 1369:1450 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 0 21 theme mass 118:121 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 6 22 theme approved 1120:1127 arg1 mAbs 1129:1132 approved mAbs 1120:1132 approved mAbs 1120:1132 CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. 25484058 9 23 theme Other 1714:1718 arg1 modifications 1738:1750 Other posttranslational modifications 1714:1750 Other posttranslational modifications hotspots 1714:1759 Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. 25484058 3 24 theme products 481:488 arg1 approval 458:465 approval 458:465 approval of biosimilar products 458:488 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 5 25 theme zone 840:843 arg1 electrophoresis 845:859 capillary zone electrophoresis 830:859 capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) 830:924 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 0 26 theme transient 53:61 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 9 27 theme modifications 1738:1750 arg1 hotspots 1752:1759 Other posttranslational modifications hotspots 1714:1759 Other posttranslational modifications hotspots 1714:1759 Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. 25484058 8 28 theme different 1695:1703 arg1 glycans 1705:1711 up to 16 different glycans 1686:1711 up to 16 different glycans 1686:1711 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 0 29 theme Monoclonal 0:9 arg1 antibodies 11:20 Monoclonal antibodies 0:20 Monoclonal antibodies 0:20 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 6 30 theme recent 1210:1215 arg1 directives 1228:1237 recent regulation directives 1210:1237 recent regulation directives 1210:1237 CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. 25484058 3 31 theme first 507:511 arg1 antibodies 524:533 the first biosimilar antibodies 503:533 the first biosimilar antibodies 503:533 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 7 32 theme 100 1314:1316 arg1 % 1317:1317 % 1317:1317 % 1317:1317 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 5 33 used used 930:933 arg2 electrophoresis 845:859 capillary zone electrophoresis 830:859 capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) 830:924 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 5 34 theme structure 1071:1079 arg1 facets 1047:1052 different facets 1037:1052 different facets of their primary structure 1037:1079 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 8 35 theme fragmentation 1546:1558 arg1 spectra 1560:1566 fragmentation spectra 1546:1566 fragmentation spectra 1546:1566 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 9 36 theme relative 1792:1799 arg1 levels 1812:1817 their relative occurrence levels 1786:1817 their relative occurrence levels 1786:1817 Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. 25484058 3 37 theme regulatory 433:442 arg1 framework 444:452 the regulatory framework 429:452 the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies 429:533 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 5 38 theme sheathless 895:904 arg1 CESI-MS 917:923 CESI-MS 917:923 CESI-MS 917:923 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 5 38 theme sheathless 895:904 arg1 interface 906:914 a sheathless interface 893:914 a sheathless interface (CESI-MS) 893:924 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 8 39 theme semiquantitative 1583:1598 arg1 analysis 1600:1607 glycoforms semiquantitative analysis 1572:1607 glycoforms semiquantitative analysis 1572:1607 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 10 40 theme using 1897:1901 arg1 CESI-MS 1903:1909 using CESI-MS 1897:1909 using CESI-MS 1897:1909 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 1 41 theme therapeutic 245:255 arg1 potency 257:263 their strong therapeutic potency 232:263 their strong therapeutic potency 232:263 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 10 42 theme ionization 1950:1959 arg1 efficiency 1961:1970 ionization efficiency 1950:1970 ionization efficiency 1950:1970 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 4 43 theme extensive 637:645 arg1 glycoproteins 573:585 highly complex glycoproteins 558:585 highly complex glycoproteins with a wide range of micro-variants 558:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 4 43 theme extensive 637:645 arg1 characterization 647:662 extensive characterization 637:662 extensive characterization 637:662 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 8 44 dep 16 1692:1693 arg1 to 1689:1690 to 1689:1690 to 1689:1690 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 0 45 theme capillary 80:88 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 10 46 theme separation 1923:1932 arg1 selectivity 1934:1944 separation selectivity 1923:1944 separation selectivity 1923:1944 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 4 47 theme micro-variants 608:621 arg1 range 599:603 a wide range 592:603 a wide range of micro-variants 592:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 7 48 theme sample 1260:1265 arg1 injection 1267:1275 only a single sample injection 1246:1275 only a single sample injection of 200 fmol 1246:1287 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 2 49 theme top-selling 296:306 arg1 drugs 308:312 the 10 top-selling drugs 289:312 the 10 top-selling drugs 289:312 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 5 50 theme marketed 951:958 arg1 mAbs 970:973 marketed reference mAbs 951:973 marketed reference mAbs 951:973 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 0 51 theme isotachophoresis 63:78 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 10 52 dep workflow 2051:2058 arg1 robustness 2060:2069 robustness 2060:2069 robustness 2060:2069 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 5 53 theme mass 872:875 arg1 spectrometry 877:888 mass spectrometry 872:888 mass spectrometry 872:888 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 10 54 theme substantial 2003:2013 arg1 improvement 2015:2025 substantial improvement 2003:2025 substantial improvement in the characterization workflow robustness and accuracy 2003:2082 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 1 55 theme most 211:214 arg1 interest 216:223 the most interest 207:223 the most interest due to their strong therapeutic potency and specificity 207:279 Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. 25484058 4 56 theme manufacturing 735:747 arg1 control 749:755 manufacturing control and biosimilarity studies 735:781 control 749:755 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 6 57 theme CESI-MS/MS 1082:1091 arg1 data 1093:1096 CESI-MS/MS data 1082:1096 CESI-MS/MS data 1082:1096 CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. 25484058 4 58 theme analytical 681:690 arg1 methods 692:698 multiple analytical methods 672:698 multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming 672:821 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 10 59 theme characterization 2034:2049 arg1 workflow 2051:2058 the characterization workflow robustness and accuracy 2030:2082 workflow 2051:2058 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 5 60 theme different 1037:1045 arg1 facets 1047:1052 different facets 1037:1052 different facets of their primary structure 1037:1079 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 8 61 theme developed 1654:1662 arg1 methodology 1664:1674 the developed methodology 1650:1674 the developed methodology 1650:1674 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 2 62 theme patent 361:366 arg1 protection 368:377 patent protection 361:377 patent protection 361:377 Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. 25484058 0 63 theme electrophoresis-tandem 95:116 arg1 spectrometry 123:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry 53:134 Monoclonal antibodies biosimilarity assessment using transient isotachophoresis capillary zone electrophoresis-tandem mass spectrometry. 25484058 7 64 theme sequence 1336:1343 arg1 characterization 1345:1360 100% amino acids (AA) sequence characterization 1314:1360 100% amino acids (AA) sequence characterization 1314:1360 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 5 65 theme biosimilar 996:1005 arg1 candidate 1007:1015 their respective biosimilar candidate 979:1015 their respective biosimilar candidate 979:1015 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 5 66 theme capillary 830:838 arg1 electrophoresis 845:859 capillary zone electrophoresis 830:859 capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) 830:924 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 9 67 theme posttranslational 1720:1736 arg1 modifications 1738:1750 Other posttranslational modifications 1714:1750 Other posttranslational modifications hotspots 1714:1759 Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. 25484058 10 68 from improvement 2015:2025 arg1 workflow 2051:2058 the characterization workflow robustness and accuracy 2030:2082 workflow 2051:2058 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 10 68 from improvement 2015:2025 arg1 accuracy 2075:2082 accuracy 2075:2082 accuracy 2075:2082 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 10 69 dep selectivity 1934:1944 arg1 the 1919:1921 the 1919:1921 the 1919:1921 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 7 70 theme acids 1325:1329 arg1 characterization 1345:1360 100% amino acids (AA) sequence characterization 1314:1360 100% amino acids (AA) sequence characterization 1314:1360 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 7 71 theme % 1317:1317 arg1 AA 1332:1333 AA 1332:1333 AA 1332:1333 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 7 71 theme % 1317:1317 arg1 acids 1325:1329 100% amino acids 1314:1329 100% amino acids (AA) sequence characterization 1314:1360 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 6 72 theme regulation 1217:1226 arg1 directives 1228:1237 recent regulation directives 1210:1237 recent regulation directives 1210:1237 CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. 25484058 4 73 theme wide 594:597 arg1 range 599:603 a wide range 592:603 a wide range of micro-variants 592:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 3 74 dep approval 458:465 arg1 antibodies 524:533 the first biosimilar antibodies 503:533 the first biosimilar antibodies 503:533 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 3 75 theme biosimilar 470:479 arg1 products 481:488 biosimilar products 470:488 biosimilar products 470:488 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 5 76 theme primary 1063:1069 arg1 structure 1071:1079 their primary structure 1057:1079 their primary structure 1057:1079 Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. 25484058 11 77 theme biosimilar 2210:2219 arg1 pipeline 2233:2240 the biosimilar development pipeline 2206:2240 the biosimilar development pipeline 2206:2240 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 3 78 theme biosimilar 513:522 arg1 antibodies 524:533 the first biosimilar antibodies 503:533 the first biosimilar antibodies 503:533 The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. 25484058 7 79 theme CESI-MS/MS 1290:1299 arg1 data 1301:1304 CESI-MS/MS data 1290:1304 CESI-MS/MS data 1290:1304 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 4 80 theme complex 565:571 arg1 glycoproteins 573:585 highly complex glycoproteins 558:585 highly complex glycoproteins with a wide range of micro-variants 558:621 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 4 80 theme complex 565:571 arg1 characterization 647:662 extensive characterization 637:662 extensive characterization 637:662 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 11 81 theme candidate 2171:2179 arg1 injection 2153:2161 a single injection 2144:2161 a single injection of each candidate enabling improvements in the biosimilar development pipeline 2144:2240 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 7 82 theme fmol 1284:1287 arg1 injection 1267:1275 only a single sample injection 1246:1275 only a single sample injection of 200 fmol 1246:1287 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 9 83 theme occurrence 1801:1810 arg1 levels 1812:1817 their relative occurrence levels 1786:1817 their relative occurrence levels 1786:1817 Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. 25484058 7 84 theme AA 1401:1402 arg1 difference 1378:1387 a difference 1376:1387 a difference of even one AA between 2 samples 1376:1420 Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. 25484058 10 85 theme CESI-MS 1903:1909 arg1 value 1888:1892 the value 1884:1892 the value of using CESI-MS 1884:1909 These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. 25484058 4 86 dep control 749:755 arg1 time-consuming 808:821 time-consuming 808:821 time-consuming 808:821 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 4 86 dep control 749:755 arg1 product 796:802 product 796:802 product 796:802 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 25484058 8 87 theme glycoforms 1572:1581 arg1 analysis 1600:1607 glycoforms semiquantitative analysis 1572:1607 glycoforms semiquantitative analysis 1572:1607 Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. 25484058 11 88 theme Biosimilarity 2085:2097 arg1 assessment 2099:2108 Biosimilarity assessment 2085:2108 Biosimilarity assessment 2085:2108 Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline. 25484058 4 89 theme multiple 672:679 arg1 methods 692:698 multiple analytical methods 672:698 multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming 672:821 As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. 24689986 6 0 theme gastric 894:900 arg1 mucosa 902:907 the canine gastric mucosa 883:907 the canine gastric mucosa 883:907 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 4 1 theme Helicobacter 570:581 arg1 species 583:589 different gastric Helicobacter species 552:589 different gastric Helicobacter species 552:589 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 7 2 theme structures 1101:1110 arg1 antigen 1118:1124 A antigen 1116:1124 A antigen 1116:1124 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 7 2 theme structures 1101:1110 arg1 expression 1080:1089 a broad expression 1072:1089 a broad expression of type 2 structures 1072:1110 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 7 2 theme structures 1101:1110 arg1 both 1127:1130 both 1127:1130 both 1127:1130 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 9 3 theme canine 1377:1382 arg1 mucosa 1392:1397 The canine gastric mucosa 1373:1397 The canine gastric mucosa 1373:1397 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 2 4 theme gastric 167:173 arg1 mucosa 175:180 The gastric mucosa 163:180 The gastric mucosa of dogs 163:188 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 5 5 theme glycosylation 657:669 arg1 profile 671:677 The glycosylation profile 653:677 The glycosylation profile 653:677 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 11 6 theme canine 1635:1640 arg1 mucosa 1650:1655 canine gastric mucosa 1635:1655 canine gastric mucosa 1635:1655 Helicobacter pylori and NHPH strains differ in their ability to adhere to canine gastric mucosa. 24689986 6 7 theme pylori 864:869 arg1 NHPH 875:878 NHPH 875:878 NHPH to the canine gastric mucosa 875:907 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 6 7 theme pylori 864:869 arg1 capacity 836:843 The in vitro binding capacity 815:843 The in vitro binding capacity of FITC-labeled H. pylori 815:869 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 6 8 dep in 819:820 arg1 vitro 822:826 vitro 822:826 vitro 822:826 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 2 9 dep H. 264:265 arg1 pylori 267:272 H. pylori 264:272 H. pylori 264:272 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 9 10 from mucosa 1463:1468 arg1 different 1430:1438 different 1430:1438 different 1430:1438 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 8 11 dep H. 1209:1210 arg1 heilmannii 1212:1221 H. heilmannii 1209:1221 H. heilmannii s.s. 1209:1226 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 2 12 theme gastric 293:299 arg1 species 314:320 the predominant gastric Helicobacter species 277:320 the predominant gastric Helicobacter species in humans 277:330 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 2 12 theme gastric 293:299 arg1 H. 264:265 H. 264:265 H. 264:265 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 6 13 theme representative 931:944 arg1 cases 925:929 cases 925:929 cases representative of the canine glycosylation pattern 925:980 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 9 14 dep CONCLUSIONS 1361:1371 arg1 showed 1399:1404 showed 1399:1404 showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp 1399:1549 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 8 15 theme highest 1242:1248 arg1 score 1259:1263 the highest adhesion score 1238:1263 the highest adhesion score 1238:1263 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 7 16 theme Lewis 1044:1048 arg1 antigens 1050:1057 type 1 Lewis antigens 1037:1057 type 1 Lewis antigens 1037:1057 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 7 17 theme gastric 1002:1008 arg1 mucosa 1010:1015 The canine gastric mucosa 991:1015 The canine gastric mucosa 991:1015 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 9 18 theme human 1449:1453 arg1 mucosa 1463:1468 the human gastric mucosa 1445:1468 the human gastric mucosa 1445:1468 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 8 19 theme antral 1194:1199 arg1 mucosa 1201:1206 the canine antral mucosa 1183:1206 the canine antral mucosa 1183:1206 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 7 20 theme type 1037:1040 arg1 antigens 1050:1057 type 1 Lewis antigens 1037:1057 type 1 Lewis antigens 1037:1057 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 2 21 theme pylori 229:234 arg1 NHPH 251:254 NHPH 251:254 NHPH 251:254 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 2 21 theme pylori 229:234 arg1 helicobacters 236:248 non-Helicobacter pylori helicobacters 212:248 non-Helicobacter pylori helicobacters (NHPH) 212:255 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 1 22 theme defined 120:126 arg1 glycophenotype 136:149 defined gastric glycophenotype 120:149 defined gastric glycophenotype 120:149 Binding to canine gastric mucosa with defined gastric glycophenotype. 24689986 12 23 with agreement 1736:1744 arg1 colonization 1764:1775 its reported colonization 1751:1775 its reported colonization of the canine stomach 1751:1797 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 8 24 theme body 1280:1283 arg1 region 1285:1290 the body region 1276:1290 the body region 1276:1290 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 9 25 theme glycan 1498:1503 arg1 receptors 1505:1513 alternative glycan receptors 1486:1513 alternative glycan receptors 1486:1513 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 7 26 theme type 1094:1097 arg1 structures 1101:1110 type 2 structures 1094:1110 type 2 structures 1094:1110 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 3 27 theme human 357:361 arg1 mucosa 371:376 the human gastric mucosa 353:376 the human gastric mucosa 353:376 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 4 28 theme gastric 490:496 arg1 mucosa 498:503 the canine gastric mucosa glycophenotype 479:518 the canine gastric mucosa glycophenotype 479:518 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 3 29 dep H. 381:382 arg1 pylori 384:389 H. pylori 381:389 H. pylori 381:389 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 8 30 theme H. 1310:1311 arg1 strain 1320:1325 the SabA-positive H. pylori strain 1292:1325 the SabA-positive H. pylori strain 1292:1325 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 8 30 theme H. 1310:1311 arg1 strain 1335:1340 the strain that adhered more 1331:1358 the strain that adhered more 1331:1358 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 5 31 theme gastric 725:731 arg1 mucosa 733:738 the canine gastric mucosa 714:738 the canine gastric mucosa 714:738 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 6 32 theme glycosylation 960:972 arg1 pattern 974:980 the canine glycosylation pattern 949:980 the canine glycosylation pattern 949:980 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 7 33 theme glandular 1151:1159 arg1 epithelium 1161:1170 the surface and glandular epithelium 1135:1170 epithelium 1161:1170 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 9 34 theme Helicobacter 1534:1545 arg1 spp 1547:1549 Helicobacter spp 1534:1549 Helicobacter spp 1534:1549 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 7 35 theme surface 1139:1145 arg1 epithelium 1161:1170 the surface and glandular epithelium 1135:1170 epithelium 1161:1170 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 1 36 theme canine 93:98 arg1 mucosa 108:113 canine gastric mucosa 93:113 canine gastric mucosa with defined gastric glycophenotype 93:149 Binding to canine gastric mucosa with defined gastric glycophenotype. 24689986 3 37 theme mucosa 371:376 arg1 colonization 337:348 The colonization 333:348 The colonization of the human gastric mucosa by H. pylori 333:389 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 3 37 theme mucosa 371:376 arg1 dependent 401:409 dependent 401:409 dependent 401:409 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 6 38 theme H. 861:862 arg1 pylori 864:869 FITC-labeled H. pylori 848:869 FITC-labeled H. pylori 848:869 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 2 39 theme dogs 185:188 arg1 mucosa 175:180 The gastric mucosa 163:180 The gastric mucosa of dogs 163:188 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 7 40 dep RESULTS 983:989 arg1 lacks 1017:1021 lacks 1017:1021 lacks expression of type 1 Lewis antigens 1017:1057 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 7 40 dep RESULTS 983:989 arg1 presents 1063:1070 presents 1063:1070 presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium 1063:1170 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 3 41 theme glycan 438:443 arg1 receptors 445:453 host glycan receptors 433:453 host glycan receptors 433:453 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 0 42 theme pylori 29:34 arg1 comparison 2:11 A comparison 0:11 A comparison of Helicobacter pylori and non-Helicobacter pylori Helicobacter 0:75 A comparison of Helicobacter pylori and non-Helicobacter pylori Helicobacter spp. 24689986 0 43 dep non-Helicobacter 40:55 arg1 Helicobacter 64:75 non-Helicobacter pylori Helicobacter 40:75 non-Helicobacter pylori Helicobacter 40:75 A comparison of Helicobacter pylori and non-Helicobacter pylori Helicobacter spp. 24689986 12 44 theme stomach 1791:1797 arg1 colonization 1764:1775 its reported colonization 1751:1775 its reported colonization of the canine stomach 1751:1797 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 4 45 theme glycophenotype 505:518 arg1 mucosa 498:503 the canine gastric mucosa glycophenotype 479:518 the canine gastric mucosa glycophenotype 479:518 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 12 46 attach presented 1693:1701 arg1 agreement 1736:1744 agreement 1736:1744 agreement with its reported colonization of the canine stomach 1736:1797 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 12 46 attach presented 1693:1701 arg2 s.s. 1688:1691 s.s. 1688:1691 s.s. 1688:1691 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 11 47 theme NHPH 1585:1588 arg1 strains 1590:1596 Helicobacter pylori and NHPH strains 1561:1596 strains 1590:1596 Helicobacter pylori and NHPH strains differ in their ability to adhere to canine gastric mucosa. 24689986 5 48 theme group 785:789 arg1 antigens 791:798 histo-blood group antigens 773:798 histo-blood group antigens 773:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 6 49 theme binding 828:834 arg1 capacity 836:843 The in vitro binding capacity 815:843 The in vitro binding capacity of FITC-labeled H. pylori 815:869 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 5 50 with MATERIALS 631:639 arg1 focus 746:750 focus 746:750 focus on the expression of histo-blood group antigens 746:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 6 51 theme in 819:820 arg1 capacity 836:843 The in vitro binding capacity 815:843 The in vitro binding capacity of FITC-labeled H. pylori 815:869 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 8 52 from strain 1335:1340 arg1 region 1285:1290 the body region 1276:1290 the body region 1276:1290 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 6 53 theme canine 887:892 arg1 mucosa 902:907 the canine gastric mucosa 883:907 the canine gastric mucosa 883:907 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 5 54 with METHODS 645:651 arg1 focus 746:750 focus 746:750 focus on the expression of histo-blood group antigens 746:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 4 55 theme gastric 562:568 arg1 species 583:589 different gastric Helicobacter species 552:589 different gastric Helicobacter species 552:589 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 2 56 theme predominant 281:291 arg1 species 314:320 the predominant gastric Helicobacter species 277:320 the predominant gastric Helicobacter species in humans 277:330 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 2 56 theme predominant 281:291 arg1 H. 264:265 H. 264:265 H. 264:265 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 9 57 theme gastric 1384:1390 arg1 mucosa 1392:1397 The canine gastric mucosa 1373:1397 The canine gastric mucosa 1373:1397 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 4 58 theme species 583:589 arg1 capacity 540:547 the capacity 536:547 the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa 536:628 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 7 59 theme broad 1074:1078 arg1 expression 1080:1089 a broad expression 1072:1089 a broad expression of type 2 structures 1072:1110 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 0 60 theme non-Helicobacter 40:55 arg1 comparison 2:11 A comparison 0:11 A comparison of Helicobacter pylori and non-Helicobacter pylori Helicobacter 0:75 A comparison of Helicobacter pylori and non-Helicobacter pylori Helicobacter spp. 24689986 12 61 theme highest 1707:1713 arg1 capacity 1724:1731 the highest adhesion capacity 1703:1731 the highest adhesion capacity 1703:1731 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 11 62 theme gastric 1642:1648 arg1 mucosa 1650:1655 canine gastric mucosa 1635:1655 canine gastric mucosa 1635:1655 Helicobacter pylori and NHPH strains differ in their ability to adhere to canine gastric mucosa. 24689986 9 63 theme glycosylation 1408:1420 arg1 profile 1422:1428 a glycosylation profile 1406:1428 a glycosylation profile different from the human gastric mucosa 1406:1468 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 1 64 with mucosa 108:113 arg1 glycophenotype 136:149 defined gastric glycophenotype 120:149 defined gastric glycophenotype 120:149 Binding to canine gastric mucosa with defined gastric glycophenotype. 24689986 6 65 theme canine 953:958 arg1 pattern 974:980 the canine glycosylation pattern 949:980 the canine glycosylation pattern 949:980 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 8 66 theme H. 1209:1210 arg1 s.s. 1223:1226 H. heilmannii s.s. 1209:1226 H. heilmannii s.s. 1209:1226 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 7 67 theme antigens 1050:1057 arg1 expression 1023:1032 expression 1023:1032 expression of type 1 Lewis antigens 1023:1057 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 2 68 theme Helicobacter 301:312 arg1 species 314:320 the predominant gastric Helicobacter species 277:320 the predominant gastric Helicobacter species in humans 277:330 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 2 68 theme Helicobacter 301:312 arg1 H. 264:265 H. 264:265 H. 264:265 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 9 69 theme different 1430:1438 arg1 profile 1422:1428 a glycosylation profile 1406:1428 a glycosylation profile different from the human gastric mucosa 1406:1468 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 4 70 theme gastric 615:621 arg1 mucosa 623:628 the canine gastric mucosa 604:628 the canine gastric mucosa 604:628 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 8 71 dep H. 1310:1311 arg1 pylori 1313:1318 H. pylori 1310:1318 the SabA-positive H. pylori strain 1292:1325 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 9 72 theme gastric 1455:1461 arg1 mucosa 1463:1468 the human gastric mucosa 1445:1468 the human gastric mucosa 1445:1468 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 8 73 theme canine 1187:1192 arg1 mucosa 1201:1206 the canine antral mucosa 1183:1206 the canine antral mucosa 1183:1206 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 5 74 theme antral 691:696 arg1 compartments 698:709 antral compartments 691:709 antral compartments 691:709 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 8 75 theme adhesion 1250:1257 arg1 score 1259:1263 the highest adhesion score 1238:1263 the highest adhesion score 1238:1263 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 12 76 theme adhesion 1715:1722 arg1 capacity 1724:1731 the highest adhesion capacity 1703:1731 the highest adhesion capacity 1703:1731 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 7 77 theme canine 995:1000 arg1 mucosa 1010:1015 The canine gastric mucosa 991:1015 The canine gastric mucosa 991:1015 RESULTS The canine gastric mucosa lacks expression of type 1 Lewis antigens and presents a broad expression of type 2 structures and A antigen, both in the surface and glandular epithelium. 24689986 1 78 theme gastric 128:134 arg1 glycophenotype 136:149 defined gastric glycophenotype 120:149 defined gastric glycophenotype 120:149 Binding to canine gastric mucosa with defined gastric glycophenotype. 24689986 5 79 from focus 746:750 arg1 expression 759:768 the expression 755:768 the expression of histo-blood group antigens 755:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 9 80 theme alternative 1486:1496 arg1 receptors 1505:1513 alternative glycan receptors 1486:1513 alternative glycan receptors 1486:1513 CONCLUSIONS The canine gastric mucosa showed a glycosylation profile different from the human gastric mucosa suggesting that alternative glycan receptors may be involved in Helicobacter spp. 24689986 4 81 theme canine 483:488 arg1 mucosa 498:503 the canine gastric mucosa glycophenotype 479:518 the canine gastric mucosa glycophenotype 479:518 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 5 82 theme canine 718:723 arg1 mucosa 733:738 the canine gastric mucosa 714:738 the canine gastric mucosa 714:738 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 2 83 from species 314:320 arg1 humans 325:330 humans 325:330 humans 325:330 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 6 84 theme pattern 974:980 arg1 representative 931:944 representative 931:944 representative 931:944 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 3 85 theme gastric 363:369 arg1 mucosa 371:376 the human gastric mucosa 353:376 the human gastric mucosa 353:376 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 8 86 theme SabA-positive 1296:1308 arg1 strain 1320:1325 the SabA-positive H. pylori strain 1292:1325 the SabA-positive H. pylori strain 1292:1325 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 8 86 theme SabA-positive 1296:1308 arg1 strain 1335:1340 the strain that adhered more 1331:1358 the strain that adhered more 1331:1358 Regarding the canine antral mucosa, H. heilmannii s.s. presented the highest adhesion score whereas in the body region the SabA-positive H. pylori strain was the strain that adhered more. 24689986 5 87 theme mucosa 733:738 arg1 body 682:685 body 682:685 body 682:685 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 5 87 theme mucosa 733:738 arg1 compartments 698:709 antral compartments 691:709 antral compartments 691:709 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 3 88 theme host 433:436 arg1 receptors 445:453 host glycan receptors 433:453 host glycan receptors 433:453 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 5 89 theme antigens 791:798 arg1 expression 759:768 the expression 755:768 the expression of histo-blood group antigens 755:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 2 90 dep BACKGROUND 152:161 arg1 colonized 199:207 colonized 199:207 colonized 199:207 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 5 91 from MATERIALS 631:639 arg1 body 682:685 body 682:685 body 682:685 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 5 91 from MATERIALS 631:639 arg1 compartments 698:709 antral compartments 691:709 antral compartments 691:709 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 1 92 theme gastric 100:106 arg1 mucosa 108:113 canine gastric mucosa 93:113 canine gastric mucosa with defined gastric glycophenotype 93:149 Binding to canine gastric mucosa with defined gastric glycophenotype. 24689986 4 93 theme different 552:560 arg1 species 583:589 different gastric Helicobacter species 552:589 different gastric Helicobacter species 552:589 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 24689986 6 94 theme FITC-labeled 848:859 arg1 pylori 864:869 FITC-labeled H. pylori 848:869 FITC-labeled H. pylori 848:869 The in vitro binding capacity of FITC-labeled H. pylori and NHPH to the canine gastric mucosa was assessed in cases representative of the canine glycosylation pattern. 24689986 5 95 dep MATERIALS 631:639 arg1 profile 671:677 The glycosylation profile 653:677 The glycosylation profile 653:677 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 3 96 theme receptors 445:453 arg1 recognition 418:428 the recognition 414:428 the recognition of host glycan receptors 414:453 The colonization of the human gastric mucosa by H. pylori is highly dependent on the recognition of host glycan receptors. 24689986 5 97 theme histo-blood 773:783 arg1 antigens 791:798 histo-blood group antigens 773:798 histo-blood group antigens 773:798 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 12 98 theme canine 1784:1789 arg1 stomach 1791:1797 the canine stomach 1780:1797 the canine stomach 1780:1797 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 2 99 theme non-Helicobacter 212:227 arg1 NHPH 251:254 NHPH 251:254 NHPH 251:254 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 2 99 theme non-Helicobacter 212:227 arg1 helicobacters 236:248 non-Helicobacter pylori helicobacters 212:248 non-Helicobacter pylori helicobacters (NHPH) 212:255 BACKGROUND The gastric mucosa of dogs is often colonized by non-Helicobacter pylori helicobacters (NHPH), while H. pylori is the predominant gastric Helicobacter species in humans. 24689986 11 100 theme Helicobacter 1561:1572 arg1 pylori 1574:1579 Helicobacter pylori and NHPH strains 1561:1596 pylori 1574:1579 Helicobacter pylori and NHPH strains differ in their ability to adhere to canine gastric mucosa. 24689986 5 101 from METHODS 645:651 arg1 body 682:685 body 682:685 body 682:685 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 5 101 from METHODS 645:651 arg1 compartments 698:709 antral compartments 691:709 antral compartments 691:709 MATERIALS AND METHODS The glycosylation profile in body and antral compartments of the canine gastric mucosa, with focus on the expression of histo-blood group antigens was evaluated. 24689986 12 102 theme reported 1755:1762 arg1 colonization 1764:1775 its reported colonization 1751:1775 its reported colonization of the canine stomach 1751:1797 Among the NHPH, H. heilmannii s.s. presented the highest adhesion capacity in agreement with its reported colonization of the canine stomach. 24689986 4 103 theme canine 608:613 arg1 mucosa 623:628 the canine gastric mucosa 604:628 the canine gastric mucosa 604:628 Our goal was to define the canine gastric mucosa glycophenotype and to evaluate the capacity of different gastric Helicobacter species to adhere to the canine gastric mucosa. 27721040 0 0 theme binding 77:83 arg1 activity 85:92 its FcγRI binding activity 67:92 its FcγRI binding activity 67:92 Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity. 27721040 8 1 with treatment 1303:1311 arg1 swainsonine 1318:1328 swainsonine 1318:1328 swainsonine which produced hybrid glycans 1318:1358 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 3 2 theme affinity 471:478 arg1 receptor 486:493 the high affinity FcγRI receptor 462:493 the high affinity FcγRI receptor 462:493 However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. 27721040 0 3 theme FcγRI 71:75 arg1 activity 85:92 its FcγRI binding activity 67:92 its FcγRI binding activity 67:92 Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity. 27721040 3 4 theme FcγRI 480:484 arg1 receptor 486:493 the high affinity FcγRI receptor 462:493 the high affinity FcγRI receptor 462:493 However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. 27721040 6 5 theme circular 932:939 arg1 dichroism 941:949 circular dichroism 932:949 circular dichroism 932:949 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 6 6 theme mAbs 1058:1061 arg1 solution-behaviour 1032:1049 the solution-behaviour 1028:1049 the solution-behaviour of the mAbs 1028:1061 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 0 7 from Inhibition 0:9 arg1 antibody 41:48 a camelid antibody 31:48 a camelid antibody 31:48 Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity. 27721040 10 8 theme mAb 1693:1695 arg1 candidate 1697:1705 a highly suitable mAb candidate 1675:1705 a highly suitable mAb candidate for therapeutic applications 1675:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 4 9 dep IgG1 656:659 arg1 DP-12 662:666 DP-12 662:666 DP-12 662:666 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 4 9 dep IgG1 656:659 arg1 150kDa 669:674 150kDa 669:674 150kDa 669:674 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 5 10 theme mannose 881:887 arg1 structures 896:905 hybrid and high mannose glycan structures 865:905 structures 896:905 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 4 11 theme hybrid 572:577 arg1 mAb 579:581 a human-llama hybrid mAb 558:581 a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa) 558:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 2 12 theme modifications 269:281 arg1 effects 245:251 The effects 241:251 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity 241:353 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 10 13 theme suitable 1684:1691 arg1 candidate 1697:1705 a highly suitable mAb candidate 1675:1705 a highly suitable mAb candidate for therapeutic applications 1675:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 1 14 theme common 131:136 arg1 practice 138:145 common practice 131:145 common practice 131:145 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 5 15 theme cell 839:842 arg1 cultures 844:851 Chinese hamster ovary (CHO) cell cultures 811:851 Chinese hamster ovary (CHO) cell cultures 811:851 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 2 16 theme glycan 262:267 arg1 modifications 269:281 these glycan modifications 256:281 these glycan modifications 256:281 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 5 17 theme Chinese 811:817 arg1 CHO 834:836 CHO 834:836 CHO 834:836 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 17 theme Chinese 811:817 arg1 ovary 827:831 Chinese hamster ovary 811:831 Chinese hamster ovary (CHO) cell cultures 811:851 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 4 18 theme human-llama 560:570 arg1 mAb 579:581 a human-llama hybrid mAb 558:581 a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa) 558:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 10 19 theme glycan 1650:1655 arg1 changes 1657:1663 glycan changes 1650:1663 glycan changes making it a highly suitable mAb candidate for therapeutic applications 1650:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 5 20 dep inhibitors 735:744 arg1 kifunensine 781:791 kifunensine 781:791 kifunensine 781:791 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 20 dep inhibitors 735:744 arg1 swainsonine 747:757 swainsonine 747:757 swainsonine 747:757 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 20 dep inhibitors 735:744 arg1 inhibitors 735:744 three glycosylation inhibitors 715:744 three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) 715:792 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 20 dep inhibitors 735:744 arg1 castanospermine 760:774 castanospermine 760:774 castanospermine 760:774 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 9 21 theme binding 1392:1398 arg1 affinity 1400:1407 the binding affinity 1388:1407 the binding affinity for EG2-hFc 1388:1419 Following de-glycosylation the binding affinity for EG2-hFc was only marginally reduced (6-fold) compared to a drastic (118-fold) decrease for DP-12. 27721040 10 22 theme enhanced 1623:1630 arg1 stability 1632:1640 its enhanced stability 1619:1640 its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications 1619:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 7 23 theme FcγRI 1197:1201 arg1 interaction 1182:1192 the interaction 1178:1192 the interaction of FcγRI with variously glycosylated mAbs 1178:1234 However, changes were observed when studying the interaction of FcγRI with variously glycosylated mAbs. 27721040 5 24 theme hybrid 865:870 arg1 structures 896:905 hybrid and high mannose glycan structures 865:905 structures 896:905 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 1 25 theme ideal 175:179 arg1 candidate 185:193 the ideal mAb candidate 171:193 the ideal mAb candidate for a wide range of therapeutic applications 171:238 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 6 26 theme Biophysical 908:918 arg1 analysis 920:927 Biophysical analysis 908:927 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation 908:1011 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 1 27 theme mAb 181:183 arg1 candidate 185:193 the ideal mAb candidate 171:193 the ideal mAb candidate for a wide range of therapeutic applications 171:238 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 10 28 theme molecular 1564:1572 arg1 weight 1574:1579 the relatively low molecular weight 1545:1579 the relatively low molecular weight of chimeric EG2-hFc 1545:1599 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 8 29 theme binding 1281:1287 arg1 affinity 1289:1296 a decreased binding affinity 1269:1296 a decreased binding affinity 1269:1296 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 8 30 theme decreased 1271:1279 arg1 affinity 1289:1296 a decreased binding affinity 1269:1296 a decreased binding affinity 1269:1296 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 10 31 dep changes 1657:1663 arg1 making 1665:1670 making 1665:1670 making it a highly suitable mAb candidate for therapeutic applications 1665:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 2 32 theme binding 290:296 arg1 affinity 298:305 the binding affinity 286:305 the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity 286:353 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 0 33 theme glycosylation 14:26 arg1 Inhibition 0:9 Inhibition 0:9 Inhibition of glycosylation on a camelid antibody 0:48 Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity. 27721040 4 34 theme N-glycosylation 539:553 arg1 effect 520:525 the effect 516:525 the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa) 516:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 8 35 theme hybrid 1345:1350 arg1 glycans 1352:1358 hybrid glycans 1345:1358 hybrid glycans 1345:1358 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 4 36 theme full-sized 645:654 arg1 IgG1 656:659 a full-sized IgG1 643:659 a full-sized IgG1 (DP-12, 150kDa) 643:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 10 37 theme therapeutic 1711:1721 arg1 applications 1723:1734 therapeutic applications 1711:1734 therapeutic applications 1711:1734 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 0 38 theme camelid 33:39 arg1 antibody 41:48 a camelid antibody 31:48 a camelid antibody 31:48 Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity. 27721040 4 39 theme variable 530:537 arg1 N-glycosylation 539:553 variable N-glycosylation 530:553 variable N-glycosylation 530:553 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 10 40 theme low 1560:1562 arg1 weight 1574:1579 the relatively low molecular weight 1545:1579 the relatively low molecular weight of chimeric EG2-hFc 1545:1599 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 2 41 from effects 245:251 arg1 affinity 298:305 the binding affinity 286:305 the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity 286:353 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 6 42 theme analytical 981:990 arg1 ultra-centrifugation 992:1011 analytical ultra-centrifugation 981:1011 analytical ultra-centrifugation 981:1011 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 7 43 gly glycosylated 1218:1229 arg1 mAbs 1231:1234 variously glycosylated mAbs 1208:1234 variously glycosylated mAbs 1208:1234 However, changes were observed when studying the interaction of FcγRI with variously glycosylated mAbs. 27721040 6 44 theme dynamic 952:958 arg1 scattering 966:975 dynamic light scattering 952:975 dynamic light scattering 952:975 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 1 45 theme wide 201:204 arg1 range 206:210 a wide range 199:210 a wide range of therapeutic applications 199:238 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 5 46 theme glycosylation 721:733 arg1 kifunensine 781:791 kifunensine 781:791 kifunensine 781:791 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 46 theme glycosylation 721:733 arg1 swainsonine 747:757 swainsonine 747:757 swainsonine 747:757 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 46 theme glycosylation 721:733 arg1 inhibitors 735:744 three glycosylation inhibitors 715:744 three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) 715:792 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 46 theme glycosylation 721:733 arg1 castanospermine 760:774 castanospermine 760:774 castanospermine 760:774 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 3 47 contain have 443:446 arg1 modifications 429:441 these modifications 423:441 these modifications 423:441 However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. 27721040 3 47 contain have 443:446 arg2 effect 411:416 the effect 407:416 the effect that these modifications have on binding to the high affinity FcγRI receptor 407:493 However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. 27721040 9 48 dep reduced 1441:1447 arg1 6-fold 1450:1455 6-fold 1450:1455 6-fold 1450:1455 Following de-glycosylation the binding affinity for EG2-hFc was only marginally reduced (6-fold) compared to a drastic (118-fold) decrease for DP-12. 27721040 6 49 theme multiple 1086:1093 arg1 concentrations 1095:1108 multiple concentrations 1086:1108 multiple concentrations 1086:1108 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 1 50 theme mAbs 115:118 arg1 Glycoengineering 95:110 Glycoengineering 95:110 Glycoengineering of mAbs 95:118 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 5 51 theme inhibitors 735:744 arg1 addition 703:710 the addition 699:710 the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures 699:851 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 7 52 with interaction 1182:1192 arg1 mAbs 1231:1234 variously glycosylated mAbs 1208:1234 variously glycosylated mAbs 1208:1234 However, changes were observed when studying the interaction of FcγRI with variously glycosylated mAbs. 27721040 3 53 theme high 466:469 arg1 receptor 486:493 the high affinity FcγRI receptor 462:493 the high affinity FcγRI receptor 462:493 However, little is understood about the effect that these modifications have on binding to the high affinity FcγRI receptor. 27721040 8 54 contain have 1264:1267 arg2 affinity 1289:1296 a decreased binding affinity 1269:1296 a decreased binding affinity 1269:1296 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 8 54 contain have 1264:1267 arg1 mAbs 1242:1245 Both mAbs 1237:1245 Both mAbs 1237:1245 Both mAbs were observed to have a decreased binding affinity upon treatment with swainsonine which produced hybrid glycans. 27721040 5 55 theme hamster 819:825 arg1 CHO 834:836 CHO 834:836 CHO 834:836 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 55 theme hamster 819:825 arg1 ovary 827:831 Chinese hamster ovary 811:831 Chinese hamster ovary (CHO) cell cultures 811:851 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 4 56 gly N-glycosylation 539:553 arg1 hybrid 572:577 a human-llama hybrid mAb 558:581 a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa) 558:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 2 57 theme mAbs 314:317 arg1 affinity 298:305 the binding affinity 286:305 the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity 286:353 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 5 58 theme high 876:879 arg1 structures 896:905 hybrid and high mannose glycan structures 865:905 structures 896:905 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 5 59 theme glycan 889:894 arg1 structures 896:905 hybrid and high mannose glycan structures 865:905 structures 896:905 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 2 60 theme IgG 310:312 arg1 mAbs 314:317 IgG mAbs 310:317 IgG mAbs 310:317 The effects of these glycan modifications on the binding affinity of IgG mAbs for FcγRIIIa and their cytotoxicity are well known. 27721040 6 61 theme light 960:964 arg1 scattering 966:975 dynamic light scattering 952:975 dynamic light scattering 952:975 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 9 62 theme drastic 1472:1478 arg1 decrease 1491:1498 a drastic (118-fold) decrease 1470:1498 a drastic (118-fold) decrease for DP-12 1470:1508 Following de-glycosylation the binding affinity for EG2-hFc was only marginally reduced (6-fold) compared to a drastic (118-fold) decrease for DP-12. 27721040 9 62 theme drastic 1472:1478 arg1 118-fold 1481:1488 118-fold 1481:1488 118-fold 1481:1488 Following de-glycosylation the binding affinity for EG2-hFc was only marginally reduced (6-fold) compared to a drastic (118-fold) decrease for DP-12. 27721040 4 63 dep mAb 579:581 arg1 80kDa 593:597 80kDa 593:597 80kDa 593:597 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 4 63 dep mAb 579:581 arg1 EG2-hFc 584:590 EG2-hFc 584:590 EG2-hFc 584:590 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 10 64 theme chimeric 1584:1591 arg1 EG2-hFc 1593:1599 chimeric EG2-hFc 1584:1599 chimeric EG2-hFc 1584:1599 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 5 65 theme ovary 827:831 arg1 cultures 844:851 Chinese hamster ovary (CHO) cell cultures 811:851 Chinese hamster ovary (CHO) cell cultures 811:851 This was achieved by the addition of three glycosylation inhibitors (swainsonine, castanospermine, and kifunensine) independently to Chinese hamster ovary (CHO) cell cultures to generate hybrid and high mannose glycan structures. 27721040 1 66 theme therapeutic 215:225 arg1 applications 227:238 therapeutic applications 215:238 therapeutic applications 215:238 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 27721040 10 67 theme EG2-hFc 1593:1599 arg1 weight 1574:1579 the relatively low molecular weight 1545:1579 the relatively low molecular weight of chimeric EG2-hFc 1545:1599 In summary, our data suggest that the relatively low molecular weight of chimeric EG2-hFc may contribute to its enhanced stability against glycan changes making it a highly suitable mAb candidate for therapeutic applications. 27721040 6 68 theme glycan 1114:1119 arg1 treatments 1121:1130 glycan treatments 1114:1130 glycan treatments 1114:1130 Biophysical analysis by circular dichroism, dynamic light scattering and analytical ultra-centrifugation confirmed that the solution-behaviour of the mAbs remained constant over multiple concentrations and glycan treatments. 27721040 4 69 from effect 520:525 arg1 mAb 579:581 a human-llama hybrid mAb 558:581 a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa) 558:675 This study analyzed the effect of variable N-glycosylation on a human-llama hybrid mAb (EG2-hFc, 80kDa) binding to FcγRI including a comparison to a full-sized IgG1 (DP-12, 150kDa). 27721040 7 70 theme glycosylated 1218:1229 arg1 mAbs 1231:1234 variously glycosylated mAbs 1208:1234 variously glycosylated mAbs 1208:1234 However, changes were observed when studying the interaction of FcγRI with variously glycosylated mAbs. 27721040 1 71 theme applications 227:238 arg1 range 206:210 a wide range 199:210 a wide range of therapeutic applications 199:238 Glycoengineering of mAbs has become common practice in attempts to generate the ideal mAb candidate for a wide range of therapeutic applications. 26256339 1 0 from fruits 179:184 arg1 RCP-II 157:162 A water-soluble polysaccharide namely RCP-II 119:162 A water-soluble polysaccharide namely RCP-II from raspberry fruits 119:184 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 0 1 theme raspberry 83:91 arg1 fruits 111:116 raspberry (Rubus idaeus L.) fruits 83:116 raspberry (Rubus idaeus L.) fruits 83:116 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 0 1 theme raspberry 83:91 arg1 Rubus 94:98 Rubus 94:98 Rubus 94:98 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 5 2 from effective 932:940 arg1 days 871:874 14 days 868:874 14 days of incubation 868:888 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 2 3 theme characteristic 368:381 arg1 structure 383:391 the characteristic structure 364:391 the characteristic structure of polysaccharide 364:409 RCP-II was an acidic heteropolysaccharide and the characteristic structure of polysaccharide was determined. 26256339 1 4 theme macroporous 266:276 arg1 D4020 284:288 macroporous resin D4020 and Sephadex G-100 columns 266:315 D4020 284:288 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 5 5 from determination 791:803 arg1 glycation 843:851 protein glycation 835:851 protein glycation 835:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 6 from galactose 532:540 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 4 7 theme concentration-dependent 755:777 arg1 manner 779:784 a concentration-dependent manner 753:784 a concentration-dependent manner 753:784 RCP-II presented high scavenging activity toward DPPH•, HO•, O2(•-) in a concentration-dependent manner. 26256339 3 8 from acid 485:488 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 1 9 theme resin 278:282 arg1 D4020 284:288 macroporous resin D4020 and Sephadex G-100 columns 266:315 D4020 284:288 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 5 10 theme glycation 978:986 arg1 reaction 988:995 non-enzymatic glycation reaction 964:995 non-enzymatic glycation reaction at early phase 964:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 11 theme 1.00:0.55:1.19:0.52:0.44:1.90 562:590 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 1 12 theme water-soluble 121:133 arg1 polysaccharide 135:148 A water-soluble polysaccharide 119:148 A water-soluble polysaccharide namely RCP-II from raspberry fruits 119:184 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 3 13 theme molar 547:551 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 1 14 theme raspberry 169:177 arg1 fruits 179:184 raspberry fruits 169:184 raspberry fruits 169:184 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 0 15 dep Rubus 94:98 arg1 L. 107:108 Rubus idaeus L. 94:108 Rubus idaeus L. 94:108 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 5 16 theme inhibitory 812:821 arg1 activity 823:830 the inhibitory activity 808:830 the inhibitory activity on protein glycation 808:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 17 from activity 823:830 arg1 glycation 843:851 protein glycation 835:851 protein glycation 835:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 18 from xylose 512:517 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 3 19 theme average 600:606 arg1 weight 618:623 the average molecular weight 596:623 the average molecular weight 596:623 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 3 19 theme average 600:606 arg1 Da 650:651 4013 Da 645:651 4013 Da 645:651 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 5 20 from phase 1006:1010 arg1 development 949:959 the development 945:959 the development of non-enzymatic glycation reaction at early phase 945:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 20 from phase 1006:1010 arg1 reaction 988:995 non-enzymatic glycation reaction 964:995 non-enzymatic glycation reaction at early phase 964:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 4 21 theme high 699:702 arg1 activity 715:722 high scavenging activity 699:722 high scavenging activity toward DPPH•, HO•, O2(•-) 699:748 RCP-II presented high scavenging activity toward DPPH•, HO•, O2(•-) in a concentration-dependent manner. 26256339 5 22 theme activity 823:830 arg1 determination 791:803 The determination 787:803 The determination of the inhibitory activity on protein glycation 787:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 23 theme molecular 608:616 arg1 weight 618:623 the average molecular weight 596:623 the average molecular weight 596:623 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 3 23 theme molecular 608:616 arg1 Da 650:651 4013 Da 645:651 4013 Da 645:651 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 5 24 theme incubation 879:888 arg1 days 871:874 14 days 868:874 14 days of incubation 868:888 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 1 25 theme Sephadex 294:301 arg1 columns 309:315 macroporous resin D4020 and Sephadex G-100 columns 266:315 columns 309:315 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 3 26 from glucose 520:526 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 5 27 theme non-enzymatic 964:976 arg1 reaction 988:995 non-enzymatic glycation reaction 964:995 non-enzymatic glycation reaction at early phase 964:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 0 28 theme biological 21:30 arg1 activities 32:41 biological activities 21:41 biological activities 21:41 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 1 29 theme G-100 303:307 arg1 columns 309:315 macroporous resin D4020 and Sephadex G-100 columns 266:315 columns 309:315 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 5 30 theme reaction 988:995 arg1 development 949:959 the development 945:959 the development of non-enzymatic glycation reaction at early phase 945:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 31 theme dextran 663:669 arg1 standards 671:679 dextran standards 663:679 dextran standards 663:679 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 1 32 theme complex 202:208 arg1 method 217:222 complex enzyme method 202:222 complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns 202:315 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 3 33 theme RCP-II 447:452 arg1 carbohydrate 431:442 The carbohydrate 427:442 The carbohydrate of RCP-II 427:452 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 1 34 theme enzyme 210:215 arg1 method 217:222 complex enzyme method 202:222 complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns 202:315 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 3 35 theme galacturonic 472:483 arg1 acid 485:488 galacturonic acid 472:488 galacturonic acid 472:488 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 4 36 attach presented 689:697 arg2 RCP-II 682:687 RCP-II 682:687 RCP-II 682:687 RCP-II presented high scavenging activity toward DPPH•, HO•, O2(•-) in a concentration-dependent manner. 26256339 4 36 attach presented 689:697 arg1 manner 779:784 a concentration-dependent manner 753:784 a concentration-dependent manner 753:784 RCP-II presented high scavenging activity toward DPPH•, HO•, O2(•-) in a concentration-dependent manner. 26256339 0 37 theme polysaccharide 54:67 arg1 Characterization 0:15 Characterization 0:15 Characterization 0:15 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 0 37 theme polysaccharide 54:67 arg1 activities 32:41 biological activities 21:41 biological activities 21:41 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 5 38 theme following 1029:1037 arg1 phases 1043:1048 the following two phases 1025:1048 the following two phases 1025:1048 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 39 from rhamnose 491:498 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 5 40 theme RCP-II 916:921 arg1 ability 905:911 the inhibitory ability 890:911 the inhibitory ability of RCP-II 890:921 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 40 theme RCP-II 916:921 arg1 effective 932:940 effective 932:940 effective 932:940 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 0 41 theme novel 48:52 arg1 polysaccharide 54:67 a novel polysaccharide 46:67 a novel polysaccharide 46:67 Characterization and biological activities of a novel polysaccharide isolated from raspberry (Rubus idaeus L.) fruits. 26256339 5 42 from glycation 843:851 arg1 determination 791:803 The determination 787:803 The determination of the inhibitory activity on protein glycation 787:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 43 from development 949:959 arg1 phase 1006:1010 early phase 1000:1010 early phase 1000:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 1 44 theme polysaccharide 135:148 arg1 RCP-II 157:162 A water-soluble polysaccharide namely RCP-II 119:162 A water-soluble polysaccharide namely RCP-II from raspberry fruits 119:184 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 5 45 theme early 1000:1004 arg1 phase 1006:1010 early phase 1000:1010 early phase 1000:1010 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 3 46 from arabinose 501:509 arg1 ratio 553:557 a molar ratio 545:557 a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 545:590 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 5 47 from days 871:874 arg1 ability 905:911 the inhibitory ability 890:911 the inhibitory ability of RCP-II 890:921 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 47 from days 871:874 arg1 effective 932:940 effective 932:940 effective 932:940 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 48 theme protein 835:841 arg1 glycation 843:851 protein glycation 835:851 protein glycation 835:851 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 2 49 theme acidic 332:337 arg1 RCP-II 318:323 RCP-II 318:323 RCP-II 318:323 RCP-II was an acidic heteropolysaccharide and the characteristic structure of polysaccharide was determined. 26256339 2 49 theme acidic 332:337 arg1 heteropolysaccharide 339:358 an acidic heteropolysaccharide 329:358 an acidic heteropolysaccharide 329:358 RCP-II was an acidic heteropolysaccharide and the characteristic structure of polysaccharide was determined. 26256339 5 50 theme inhibitory 894:903 arg1 ability 905:911 the inhibitory ability 890:911 the inhibitory ability of RCP-II 890:921 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 5 50 theme inhibitory 894:903 arg1 effective 932:940 effective 932:940 effective 932:940 The determination of the inhibitory activity on protein glycation showed that in 14 days of incubation the inhibitory ability of RCP-II was more effective on the development of non-enzymatic glycation reaction at early phase than that at the following two phases. 26256339 2 51 theme polysaccharide 396:409 arg1 structure 383:391 the characteristic structure 364:391 the characteristic structure of polysaccharide 364:409 RCP-II was an acidic heteropolysaccharide and the characteristic structure of polysaccharide was determined. 26256339 1 52 theme successive 236:245 arg1 purification 247:258 successive purification 236:258 successive purification using macroporous resin D4020 and Sephadex G-100 columns 236:315 A water-soluble polysaccharide namely RCP-II from raspberry fruits was obtained by complex enzyme method followed by successive purification using macroporous resin D4020 and Sephadex G-100 columns. 26256339 3 53 theme 4013 645:648 arg1 weight 618:623 the average molecular weight 596:623 the average molecular weight 596:623 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 3 53 theme 4013 645:648 arg1 Da 650:651 4013 Da 645:651 4013 Da 645:651 The carbohydrate of RCP-II was composed with galacturonic acid, rhamnose, arabinose, xylose, glucose and galactose in a molar ratio of 1.00:0.55:1.19:0.52:0.44:1.90 and the average molecular weight was estimated to be 4013 Da, based on dextran standards. 26256339 4 54 theme scavenging 704:713 arg1 activity 715:722 high scavenging activity 699:722 high scavenging activity toward DPPH•, HO•, O2(•-) 699:748 RCP-II presented high scavenging activity toward DPPH•, HO•, O2(•-) in a concentration-dependent manner. 29111696 5 0 theme sulfate 622:628 arg1 chain 634:638 the dermatan sulfate GAG chain 609:638 the dermatan sulfate GAG chain of decorin PG 609:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 7 1 theme GAG 962:964 arg1 example 949:955 the second example 938:955 the second example of a GAG with a prominent structural motif 938:998 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 2 theme sulfate 892:898 arg1 chain 900:904 the decorin dermatan sulfate chain 871:904 the decorin dermatan sulfate chain 871:904 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 5 3 theme GAG 630:632 arg1 chain 634:638 the dermatan sulfate GAG chain 609:638 the dermatan sulfate GAG chain of decorin PG 609:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 6 4 theme dermatan 883:890 arg1 chain 900:904 the decorin dermatan sulfate chain 871:904 the decorin dermatan sulfate chain 871:904 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 5 5 theme chain 634:638 arg1 structure 596:604 the much more complex structure 574:604 the much more complex structure of the dermatan sulfate GAG chain of decorin PG 574:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 3 6 theme glycoconjugates 357:371 arg1 classes 346:352 the most structurally complex classes 316:352 the most structurally complex classes of glycoconjugates 316:371 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 6 7 theme decorin 875:881 arg1 chain 900:904 the decorin dermatan sulfate chain 871:904 the decorin dermatan sulfate chain 871:904 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 2 8 theme proteomic 278:286 arg1 analysis 299:306 proteomic or genomic analysis 278:306 proteomic or genomic analysis 278:306 Glycosylation occurs in the endoplasmic reticulum and the Golgi organelle and its control is neither well-understood nor predictable based on proteomic or genomic analysis. 29111696 3 9 theme complex 338:344 arg1 classes 346:352 the most structurally complex classes 316:352 the most structurally complex classes of glycoconjugates 316:371 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 5 10 theme complex 588:594 arg1 structure 596:604 the much more complex structure 574:604 the much more complex structure of the dermatan sulfate GAG chain of decorin PG 574:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 6 11 theme separation 682:691 arg1 methods 693:699 sophisticated separation methods 668:699 sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach 668:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 3 12 theme classes 346:352 arg1 proteoglycans 380:392 the proteoglycans 376:392 the proteoglycans (PGs) 376:398 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 3 12 theme classes 346:352 arg1 classes 346:352 the most structurally complex classes 316:352 the most structurally complex classes of glycoconjugates 316:371 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 3 12 theme classes 346:352 arg1 One 309:311 One 309:311 One 309:311 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 6 13 theme mass 764:767 arg1 spectrometry 769:780 tandem mass spectrometry 757:780 tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach 757:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 7 14 with GAG 962:964 arg1 motif 994:998 a prominent structural motif 971:998 a prominent structural motif 971:998 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 15 theme sophisticated 668:680 arg1 methods 693:699 sophisticated separation methods 668:699 sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach 668:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 2 16 dep neither 229:235 arg1 well-understood 237:251 well-understood 237:251 well-understood 237:251 Glycosylation occurs in the endoplasmic reticulum and the Golgi organelle and its control is neither well-understood nor predictable based on proteomic or genomic analysis. 29111696 5 17 theme current 551:557 arg1 study 559:563 The current study 547:563 The current study 547:563 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 3 18 theme glycosaminoglycan 410:426 arg1 chains 439:444 their glycosaminoglycan (GAG) side chains 404:444 their glycosaminoglycan (GAG) side chains 404:444 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 4 19 theme bikunin 535:541 arg1 PG 543:544 the bikunin PG 531:544 the bikunin PG 531:544 Previously, our laboratory solved the structure of the chondroitin sulfate chain of the bikunin PG. 29111696 5 20 theme dermatan 613:620 arg1 sulfate 622:628 dermatan sulfate 613:628 the dermatan sulfate GAG chain of decorin PG 609:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 6 21 theme tandem 757:762 arg1 spectrometry 769:780 tandem mass spectrometry 757:780 tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach 757:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 1 22 theme last 82:85 arg1 frontiers 87:95 the last frontiers 78:95 the last frontiers 78:95 Glycomics represents one of the last frontiers and most challenging in omic analysis. 29111696 0 23 theme Sulfate 24:30 arg1 Chain 32:36 the Dermatan Sulfate Chain 11:36 the Dermatan Sulfate Chain of Decorin 11:47 Sequencing the Dermatan Sulfate Chain of Decorin. 29111696 7 24 theme second 942:947 arg1 example 949:955 the second example 938:955 the second example of a GAG with a prominent structural motif 938:998 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 1 25 theme frontiers 87:95 arg1 one 71:73 one 71:73 one 71:73 Glycomics represents one of the last frontiers and most challenging in omic analysis. 29111696 1 25 theme frontiers 87:95 arg1 frontiers 87:95 the last frontiers 78:95 the last frontiers 78:95 Glycomics represents one of the last frontiers and most challenging in omic analysis. 29111696 0 26 theme Dermatan 15:22 arg1 Chain 32:36 the Dermatan Sulfate Chain 11:36 the Dermatan Sulfate Chain of Decorin 11:47 Sequencing the Dermatan Sulfate Chain of Decorin. 29111696 7 27 theme class 1052:1056 arg1 simpler 1089:1095 simpler 1089:1095 simpler 1089:1095 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 7 27 theme class 1052:1056 arg1 variability 1032:1042 the structural variability 1017:1042 the structural variability of this class of glycoconjugates 1017:1075 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 28 theme structural 850:859 arg1 motif 861:865 the structural motif 846:865 the structural motif for the decorin dermatan sulfate chain 846:904 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 2 29 theme genomic 291:297 arg1 analysis 299:306 proteomic or genomic analysis 278:306 proteomic or genomic analysis 278:306 Glycosylation occurs in the endoplasmic reticulum and the Golgi organelle and its control is neither well-understood nor predictable based on proteomic or genomic analysis. 29111696 4 30 theme PG 543:544 arg1 chain 522:526 the chondroitin sulfate chain 498:526 the chondroitin sulfate chain of the bikunin PG 498:544 Previously, our laboratory solved the structure of the chondroitin sulfate chain of the bikunin PG. 29111696 2 31 theme endoplasmic 164:174 arg1 reticulum 176:184 the endoplasmic reticulum 160:184 the endoplasmic reticulum 160:184 Glycosylation occurs in the endoplasmic reticulum and the Golgi organelle and its control is neither well-understood nor predictable based on proteomic or genomic analysis. 29111696 3 32 theme side 434:437 arg1 chains 439:444 their glycosaminoglycan (GAG) side chains 404:444 their glycosaminoglycan (GAG) side chains 404:444 One of the most structurally complex classes of glycoconjugates is the proteoglycans (PGs) and their glycosaminoglycan (GAG) side chains. 29111696 4 33 theme sulfate 514:520 arg1 chain 522:526 the chondroitin sulfate chain 498:526 the chondroitin sulfate chain of the bikunin PG 498:544 Previously, our laboratory solved the structure of the chondroitin sulfate chain of the bikunin PG. 29111696 4 34 theme chondroitin 502:512 arg1 chain 522:526 the chondroitin sulfate chain 498:526 the chondroitin sulfate chain of the bikunin PG 498:544 Previously, our laboratory solved the structure of the chondroitin sulfate chain of the bikunin PG. 29111696 5 35 theme decorin 643:649 arg1 chain 634:638 the dermatan sulfate GAG chain 609:638 the dermatan sulfate GAG chain of decorin PG 609:652 The current study examines the much more complex structure of the dermatan sulfate GAG chain of decorin PG. 29111696 6 36 theme genetic 818:824 arg1 approach 836:843 a modified genetic algorithm approach 807:843 a modified genetic algorithm approach 807:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 7 37 theme prominent 973:981 arg1 motif 994:998 a prominent structural motif 971:998 a prominent structural motif 971:998 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 0 38 theme Decorin 41:47 arg1 Chain 32:36 the Dermatan Sulfate Chain 11:36 the Dermatan Sulfate Chain of Decorin 11:47 Sequencing the Dermatan Sulfate Chain of Decorin. 29111696 7 39 theme structural 1021:1030 arg1 simpler 1089:1095 simpler 1089:1095 simpler 1089:1095 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 7 39 theme structural 1021:1030 arg1 variability 1032:1042 the structural variability 1017:1042 the structural variability of this class of glycoconjugates 1017:1075 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 40 theme algorithm 826:834 arg1 approach 836:843 a modified genetic algorithm approach 807:843 a modified genetic algorithm approach 807:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 0 41 dep Chain 32:36 arg1 Sequencing 0:9 Sequencing 0:9 Sequencing 0:9 Sequencing the Dermatan Sulfate Chain of Decorin. 29111696 6 42 theme domain 737:742 arg1 mapping 744:750 domain mapping 737:750 domain mapping 737:750 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 4 43 theme chain 522:526 arg1 structure 485:493 the structure 481:493 the structure of the chondroitin sulfate chain of the bikunin PG 481:544 Previously, our laboratory solved the structure of the chondroitin sulfate chain of the bikunin PG. 29111696 2 44 theme Golgi 194:198 arg1 organelle 200:208 the Golgi organelle 190:208 the Golgi organelle 190:208 Glycosylation occurs in the endoplasmic reticulum and the Golgi organelle and its control is neither well-understood nor predictable based on proteomic or genomic analysis. 29111696 7 45 theme structural 983:992 arg1 motif 994:998 a prominent structural motif 971:998 a prominent structural motif 971:998 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 46 theme modified 809:816 arg1 approach 836:843 a modified genetic algorithm approach 807:843 a modified genetic algorithm approach 807:843 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 29111696 1 47 theme omic 121:124 arg1 analysis 126:133 omic analysis 121:133 omic analysis 121:133 Glycomics represents one of the last frontiers and most challenging in omic analysis. 29111696 7 48 theme glycoconjugates 1061:1075 arg1 class 1052:1056 this class 1047:1056 this class of glycoconjugates 1047:1075 This represents the second example of a GAG with a prominent structural motif, suggesting that the structural variability of this class of glycoconjugates is somewhat simpler than had been expected. 29111696 6 49 theme compositional 713:725 arg1 analysis 727:734 compositional analysis 713:734 compositional analysis 713:734 By utilizing sophisticated separation methods followed by compositional analysis, domain mapping, and tandem mass spectrometry coupled with analysis by a modified genetic algorithm approach, the structural motif for the decorin dermatan sulfate chain was determined. 27864768 3 0 theme biosynthesis 339:350 arg1 understanding 313:325 understanding 313:325 understanding of N-glycan biosynthesis 313:350 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 4 1 theme proteins 509:516 arg1 group 500:504 an interesting group 485:504 an interesting group of proteins 485:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 1 theme proteins 509:516 arg1 receptors 471:479 plant hormone receptors 457:479 plant hormone receptors 457:479 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 1 theme proteins 509:516 arg1 proteins 509:516 proteins 509:516 proteins 509:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 2 gly N-glycosylation 656:670 arg2 sites 672:676 numerous potential N-glycosylation sites 637:676 numerous potential N-glycosylation sites 637:676 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 3 theme secretory 584:592 arg1 pathway 594:600 the secretory pathway 580:600 the secretory pathway 580:600 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 6 4 theme hormone 962:968 arg1 receptors 970:978 endogenous hormone receptors 951:978 endogenous hormone receptors like the brassinosteroid receptor BRI1 951:1017 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 6 5 theme immunoblot-based 864:879 arg1 approach 881:888 a straightforward immunoblot-based approach 846:888 a straightforward immunoblot-based approach 846:888 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 3 6 theme function 379:386 arg1 knowledge 357:365 our knowledge 353:365 our knowledge of N-glycan function on individual plant proteins 353:415 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 6 7 theme endogenous 951:960 arg1 receptors 970:978 endogenous hormone receptors 951:978 endogenous hormone receptors like the brassinosteroid receptor BRI1 951:1017 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 4 8 located present 551:557 arg2 several 521:527 several 521:527 several 521:527 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 8 located present 551:557 arg1 sites 571:575 distinct sites 562:575 distinct sites in the secretory pathway 562:600 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 9 theme interesting 488:498 arg1 group 500:504 an interesting group 485:504 an interesting group of proteins 485:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 9 theme interesting 488:498 arg1 receptors 471:479 plant hormone receptors 457:479 plant hormone receptors 457:479 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 9 theme interesting 488:498 arg1 proteins 509:516 proteins 509:516 proteins 509:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 3 10 from progress 301:308 arg1 understanding 313:325 understanding 313:325 understanding of N-glycan biosynthesis 313:350 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 2 11 theme modifications 141:153 arg1 modifications 141:153 the most common covalent protein modifications 108:153 the most common covalent protein modifications 108:153 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 11 theme modifications 141:153 arg1 one 101:103 one 101:103 one 101:103 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 3 12 theme N-glycan 370:377 arg1 function 379:386 N-glycan function 370:386 N-glycan function 370:386 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 2 13 theme protein 133:139 arg1 modifications 141:153 the most common covalent protein modifications 108:153 the most common covalent protein modifications 108:153 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 4 14 from group 500:504 arg1 respect 448:454 this respect 443:454 this respect 443:454 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 1 15 theme life 76:79 arg1 trees 67:71 all trees 63:71 all trees of life 63:79 Glycosylation is essential for all trees of life. 27864768 4 16 theme N-glycosylation 656:670 arg1 sites 672:676 numerous potential N-glycosylation sites 637:676 numerous potential N-glycosylation sites 637:676 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 0 17 theme Receptor 8:15 arg1 Glycosylation 17:29 Hormone Receptor Glycosylation 0:29 Hormone Receptor Glycosylation. 0:30 Hormone Receptor Glycosylation. 27864768 2 18 theme covalent 124:131 arg1 modifications 141:153 the most common covalent protein modifications 108:153 the most common covalent protein modifications 108:153 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 6 19 theme N-glycosylation 932:946 arg1 analysis 920:927 the analysis 916:927 the analysis of N-glycosylation 916:946 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 0 20 theme Hormone 0:6 arg1 Glycosylation 17:29 Hormone Receptor Glycosylation 0:29 Hormone Receptor Glycosylation. 0:30 Hormone Receptor Glycosylation. 27864768 4 21 theme distinct 562:569 arg1 sites 571:575 distinct sites 562:575 distinct sites in the secretory pathway 562:600 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 6 22 theme straightforward 848:862 arg1 approach 881:888 a straightforward immunoblot-based approach 846:888 a straightforward immunoblot-based approach 846:888 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 4 23 theme plant 457:461 arg1 group 500:504 an interesting group 485:504 an interesting group of proteins 485:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 23 theme plant 457:461 arg1 receptors 471:479 plant hormone receptors 457:479 plant hormone receptors 457:479 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 23 theme plant 457:461 arg1 proteins 509:516 proteins 509:516 proteins 509:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 3 24 theme individual 391:400 arg1 proteins 408:415 individual plant proteins 391:415 individual plant proteins 391:415 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 4 25 from sites 571:575 arg1 pathway 594:600 the secretory pathway 580:600 the secretory pathway 580:600 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 25 from sites 571:575 arg1 present 551:557 present 551:557 present 551:557 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 3 26 theme recent 294:299 arg1 progress 301:308 recent progress 294:308 recent progress in understanding of N-glycan biosynthesis 294:350 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 3 27 theme plant 402:406 arg1 proteins 408:415 individual plant proteins 391:415 individual plant proteins 391:415 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 4 28 theme hormone 463:469 arg1 group 500:504 an interesting group 485:504 an interesting group of proteins 485:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 28 theme hormone 463:469 arg1 receptors 471:479 plant hormone receptors 457:479 plant hormone receptors 457:479 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 28 theme hormone 463:469 arg1 proteins 509:516 proteins 509:516 proteins 509:516 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 29 attach present 551:557 arg2 several 521:527 several 521:527 several 521:527 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 29 attach present 551:557 arg1 sites 571:575 distinct sites 562:575 distinct sites in the secretory pathway 562:600 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 29 attach present 551:557 arg1 membrane 619:626 the plasma membrane 608:626 the plasma membrane 608:626 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 2 30 theme large 172:176 arg1 variety 178:184 a large variety 170:184 a large variety of cellular processes including protein folding, quality control and protein-receptor interactions 170:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 30 theme large 172:176 arg1 processes 198:206 cellular processes 189:206 cellular processes including protein folding, quality control and protein-receptor interactions 189:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 30 theme large 172:176 arg1 folding 226:232 protein folding 218:232 protein folding 218:232 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 30 theme large 172:176 arg1 control 243:249 quality control 235:249 quality control 235:249 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 30 theme large 172:176 arg1 interactions 272:283 protein-receptor interactions 255:283 protein-receptor interactions 255:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 31 theme quality 235:241 arg1 control 243:249 quality control 235:249 quality control 235:249 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 4 32 theme potential 646:654 arg1 sites 672:676 numerous potential N-glycosylation sites 637:676 numerous potential N-glycosylation sites 637:676 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 5 33 theme abundant 809:816 arg1 modification 826:837 this abundant protein modification 804:837 this abundant protein modification 804:837 Identifying and characterization of N-glycan structures on these proteins is essential to investigate the functional role of this abundant protein modification. 27864768 4 34 theme proteins 538:545 arg1 several 521:527 several 521:527 several 521:527 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 35 theme numerous 637:644 arg1 sites 672:676 numerous potential N-glycosylation sites 637:676 numerous potential N-glycosylation sites 637:676 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 5 36 theme protein 818:824 arg1 modification 826:837 this abundant protein modification 804:837 this abundant protein modification 804:837 Identifying and characterization of N-glycan structures on these proteins is essential to investigate the functional role of this abundant protein modification. 27864768 6 37 theme receptor 1005:1012 arg1 BRI1 1014:1017 the brassinosteroid receptor BRI1 985:1017 the brassinosteroid receptor BRI1 985:1017 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 4 38 contain have 632:635 arg2 sites 672:676 numerous potential N-glycosylation sites 637:676 numerous potential N-glycosylation sites 637:676 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 4 38 contain have 632:635 arg1 several 521:527 several 521:527 several 521:527 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 5 39 theme modification 826:837 arg1 role 796:799 the functional role 781:799 the functional role of this abundant protein modification 781:837 Identifying and characterization of N-glycan structures on these proteins is essential to investigate the functional role of this abundant protein modification. 27864768 6 40 theme brassinosteroid 989:1003 arg1 BRI1 1014:1017 the brassinosteroid receptor BRI1 985:1017 the brassinosteroid receptor BRI1 985:1017 Here, a straightforward immunoblot-based approach is presented that enables the analysis of N-glycosylation on endogenous hormone receptors like the brassinosteroid receptor BRI1. 27864768 3 41 theme N-glycan 330:337 arg1 biosynthesis 339:350 N-glycan biosynthesis 330:350 N-glycan biosynthesis 330:350 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 2 42 theme protein 218:224 arg1 folding 226:232 protein folding 218:232 protein folding 218:232 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 43 theme processes 198:206 arg1 variety 178:184 a large variety 170:184 a large variety of cellular processes including protein folding, quality control and protein-receptor interactions 170:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 43 theme processes 198:206 arg1 processes 198:206 cellular processes 189:206 cellular processes including protein folding, quality control and protein-receptor interactions 189:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 43 theme processes 198:206 arg1 folding 226:232 protein folding 218:232 protein folding 218:232 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 43 theme processes 198:206 arg1 control 243:249 quality control 235:249 quality control 235:249 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 43 theme processes 198:206 arg1 interactions 272:283 protein-receptor interactions 255:283 protein-receptor interactions 255:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 5 44 theme N-glycan 715:722 arg1 structures 724:733 N-glycan structures 715:733 N-glycan structures 715:733 Identifying and characterization of N-glycan structures on these proteins is essential to investigate the functional role of this abundant protein modification. 27864768 2 45 theme cellular 189:196 arg1 processes 198:206 cellular processes 189:206 cellular processes including protein folding, quality control and protein-receptor interactions 189:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 45 theme cellular 189:196 arg1 folding 226:232 protein folding 218:232 protein folding 218:232 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 45 theme cellular 189:196 arg1 control 243:249 quality control 235:249 quality control 235:249 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 45 theme cellular 189:196 arg1 interactions 272:283 protein-receptor interactions 255:283 protein-receptor interactions 255:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 2 46 theme protein-receptor 255:270 arg1 interactions 272:283 protein-receptor interactions 255:283 protein-receptor interactions 255:283 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 4 47 from membrane 619:626 arg1 present 551:557 present 551:557 present 551:557 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27864768 3 48 from knowledge 357:365 arg1 proteins 408:415 individual plant proteins 391:415 individual plant proteins 391:415 Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. 27864768 5 49 theme functional 785:794 arg1 role 796:799 the functional role 781:799 the functional role of this abundant protein modification 781:837 Identifying and characterization of N-glycan structures on these proteins is essential to investigate the functional role of this abundant protein modification. 27864768 2 50 theme common 117:122 arg1 modifications 141:153 the most common covalent protein modifications 108:153 the most common covalent protein modifications 108:153 N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. 27864768 4 51 theme plasma 612:617 arg1 membrane 619:626 the plasma membrane 608:626 the plasma membrane 608:626 In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites. 27352363 3 0 from sp 412:413 arg1 RhaL1 389:393 RhaL1 389:393 RhaL1 389:393 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 3 0 from sp 412:413 arg1 α-L-rhamnosidase 371:386 a wild-type α-L-rhamnosidase 359:386 a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp 359:413 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 5 1 theme catalytic 664:672 arg1 activity 674:681 the catalytic activity 660:681 the catalytic activity of RhaL1 660:690 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 8 2 theme improved 1425:1432 arg1 due 1459:1461 due 1459:1461 due 1459:1461 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 8 2 theme improved 1425:1432 arg1 yield 1434:1438 the improved yield 1421:1438 the improved yield of mutant W261Y 1421:1454 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 9 3 theme pH-activity 1595:1605 arg1 profile 1607:1613 the pH-activity profile 1591:1613 the pH-activity profile in hydrolysis reaction 1591:1636 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 3 4 theme previous 331:338 arg1 work 340:343 Our previous work 327:343 Our previous work 327:343 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 4 5 theme glycosyl 537:544 arg1 donor 546:550 glycosyl donor 537:550 glycosyl donor 537:550 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 8 6 theme structural 1383:1392 arg1 modeling 1394:1401 the 3D structural modeling 1376:1401 the 3D structural modeling 1376:1401 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 9 7 from shift 1582:1586 arg1 profile 1607:1613 the pH-activity profile 1591:1613 the pH-activity profile in hydrolysis reaction 1591:1636 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 9 8 theme catalysis 1727:1735 arg1 behavior 1737:1744 the catalysis behavior 1723:1744 the catalysis behavior of the enzyme 1723:1758 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 3 9 theme wild-type 361:369 arg1 RhaL1 389:393 RhaL1 389:393 RhaL1 389:393 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 3 9 theme wild-type 361:369 arg1 α-L-rhamnosidase 371:386 a wild-type α-L-rhamnosidase 359:386 a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp 359:413 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 6 10 theme critical 806:813 arg1 residues 826:833 the critical amino acid residues 802:833 the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 802:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 11 dep donor 1024:1028 arg1 Trp369 1059:1064 Trp369 1059:1064 Trp369 1059:1064 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 11 dep donor 1024:1028 arg1 Tyr302 1039:1044 Tyr302 1039:1044 Tyr302 1039:1044 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 11 dep donor 1024:1028 arg1 Tyr316 1047:1052 Tyr316 1047:1052 Tyr316 1047:1052 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 8 12 theme catalytic 1521:1529 arg1 residue 1536:1542 the putative catalytic acid residue Asp257 1508:1549 the putative catalytic acid residue Asp257 1508:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 4 13 theme hydrolysis 488:497 arg1 reaction 499:506 reverse hydrolysis reaction 480:506 reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor 480:550 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 5 14 theme RhaL1 686:690 arg1 activity 674:681 the catalytic activity 660:681 the catalytic activity of RhaL1 660:690 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 5 15 theme amino 626:630 arg1 residues 637:644 the amino acid residues 622:644 the amino acid residues essential for the catalytic activity of RhaL1 622:690 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 2 16 used used 260:263 arg2 α-L-rhamnosidase 148:163 The α-L-rhamnosidase 144:163 The α-L-rhamnosidase 144:163 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 8 17 theme putative 1512:1519 arg1 residue 1536:1542 the putative catalytic acid residue Asp257 1508:1549 the putative catalytic acid residue Asp257 1508:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 0 18 from sp 62:63 arg1 mutagenesis 14:24 Site-directed mutagenesis 0:24 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp 0:63 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp. 27352363 9 19 theme Mutant 1552:1557 arg1 W261Y 1559:1563 Mutant W261Y 1552:1563 Mutant W261Y 1552:1563 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 5 20 theme acid 632:635 arg1 residues 637:644 the amino acid residues 622:644 the amino acid residues essential for the catalytic activity of RhaL1 622:690 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 6 21 theme positive 1116:1123 arg1 Y302F 1141:1145 Y302F 1141:1145 Y302F 1141:1145 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 21 theme positive 1116:1123 arg1 Y316F 1152:1156 Y316F 1152:1156 Y316F 1152:1156 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 21 theme positive 1116:1123 arg1 W261Y 1134:1138 W261Y 1134:1138 W261Y 1134:1138 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 21 theme positive 1116:1123 arg1 mutants 1125:1131 three positive mutants 1110:1131 three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood 1110:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 2 22 from glycosides 235:244 arg1 release 190:196 the hydrolytic release 175:196 the hydrolytic release of rhamnose from polysaccharides and glycosides 175:244 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 6 23 theme improved 1164:1171 arg1 stood 1187:1191 improved product yield stood 1164:1191 improved product yield stood 1164:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 2 24 from polysaccharides 215:229 arg1 release 190:196 the hydrolytic release 175:196 the hydrolytic release of rhamnose from polysaccharides and glycosides 175:244 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 7 25 theme wild-type 1349:1357 arg1 enzyme 1359:1364 the wild-type enzyme 1345:1364 the wild-type enzyme 1345:1364 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 4 26 theme inexpensive 513:523 arg1 rhamnose 525:532 inexpensive rhamnose 513:532 inexpensive rhamnose as glycosyl donor 513:550 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 5 27 theme hydrolysis 585:594 arg1 reaction 596:603 reverse hydrolysis reaction 577:603 reverse hydrolysis reaction 577:603 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 1 28 theme synthesis 80:88 arg1 yield 90:94 synthesis yield 80:94 synthesis yield of reverse hydrolysis based on rational design 80:141 L1 to enhance synthesis yield of reverse hydrolysis based on rational design. 27352363 9 29 from residue 1678:1684 arg1 cavity 1705:1710 the active site cavity 1689:1710 the active site cavity 1689:1710 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 8 30 theme W261Y 1450:1454 arg1 due 1459:1461 due 1459:1461 due 1459:1461 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 8 30 theme W261Y 1450:1454 arg1 yield 1434:1438 the improved yield 1421:1438 the improved yield of mutant W261Y 1421:1454 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 6 31 theme product 1173:1179 arg1 stood 1187:1191 improved product yield stood 1164:1191 improved product yield stood 1164:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 5 32 theme reaction 596:603 arg1 yield 568:572 the yield 564:572 the yield of reverse hydrolysis reaction 564:603 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 4 33 theme reverse 480:486 arg1 reaction 499:506 reverse hydrolysis reaction 480:506 reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor 480:550 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 7 34 theme mutant 1232:1237 arg1 W261Y 1239:1243 mutant W261Y 1232:1243 mutant W261Y 1232:1243 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 0 35 theme Site-directed 0:12 arg1 mutagenesis 14:24 Site-directed mutagenesis 0:24 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp 0:63 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp. 27352363 8 36 theme mutant 1443:1448 arg1 W261Y 1450:1454 mutant W261Y 1443:1454 mutant W261Y 1443:1454 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 4 37 theme rhamnose-containing 436:454 arg1 RCCs 467:470 RCCs 467:470 RCCs 467:470 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 4 37 theme rhamnose-containing 436:454 arg1 chemicals 456:464 rhamnose-containing chemicals 436:464 rhamnose-containing chemicals (RCCs) 436:471 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 2 38 theme processes 316:324 arg1 variety 294:300 a variety 292:300 a variety of industrial processes 292:324 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 2 38 theme processes 316:324 arg1 processes 316:324 industrial processes 305:324 industrial processes 305:324 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 7 39 theme positive 1213:1220 arg1 variants 1222:1229 the three positive variants 1203:1229 the three positive variants 1203:1229 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 0 40 theme α-L-rhamnosidase 29:44 arg1 mutagenesis 14:24 Site-directed mutagenesis 0:24 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp 0:63 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp. 27352363 6 41 dep form 845:848 arg1 Glu530 937:942 Glu530 937:942 Glu530 937:942 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 41 dep form 845:848 arg1 Asp257 921:926 Asp257 921:926 Asp257 921:926 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 41 dep form 845:848 arg1 His553 953:958 His553 953:958 His553 953:958 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 41 dep form 845:848 arg1 Trp555 965:970 Trp555 965:970 Trp555 965:970 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 42 theme acid 821:824 arg1 residues 826:833 the critical amino acid residues 802:833 the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 802:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 2 43 theme industrial 305:314 arg1 processes 316:324 industrial processes 305:324 industrial processes 305:324 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 7 44 from increase 1297:1304 arg1 yield 1327:1331 relative yield 1318:1331 relative yield 1318:1331 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 7 45 theme relative 1318:1325 arg1 yield 1327:1331 relative yield 1318:1331 relative yield 1318:1331 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 5 46 theme residues 725:732 arg1 mutagenesis 707:717 site-directed mutagenesis 693:717 site-directed mutagenesis of 11 residues 693:732 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 6 47 theme donor 897:901 arg1 rhamnose 903:910 donor rhamnose 897:910 donor rhamnose 897:910 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 1 48 theme hydrolysis 107:116 arg1 yield 90:94 synthesis yield 80:94 synthesis yield of reverse hydrolysis based on rational design 80:141 L1 to enhance synthesis yield of reverse hydrolysis based on rational design. 27352363 0 49 theme Alternaria 51:60 arg1 sp 62:63 Alternaria sp 51:63 Alternaria sp 51:63 Site-directed mutagenesis of α-L-rhamnosidase from Alternaria sp. 27352363 6 50 with mutants 1125:1131 arg1 stood 1187:1191 improved product yield stood 1164:1191 improved product yield stood 1164:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 8 51 theme 3D 1380:1381 arg1 modeling 1394:1401 the 3D structural modeling 1376:1401 the 3D structural modeling 1376:1401 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 1 52 theme reverse 99:105 arg1 hydrolysis 107:116 reverse hydrolysis 99:116 reverse hydrolysis based on rational design 99:141 L1 to enhance synthesis yield of reverse hydrolysis based on rational design. 27352363 6 53 theme yield 1181:1185 arg1 stood 1187:1191 improved product yield stood 1164:1191 improved product yield stood 1164:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 2 54 theme hydrolytic 179:188 arg1 release 190:196 the hydrolytic release 175:196 the hydrolytic release of rhamnose from polysaccharides and glycosides 175:244 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 8 55 theme position 1496:1503 arg1 adjustment 1470:1479 the adjustment 1466:1479 the adjustment of the spatial position of the putative catalytic acid residue Asp257 1466:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 6 56 dep mutants 1125:1131 arg1 Y302F 1141:1145 Y302F 1141:1145 Y302F 1141:1145 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 56 dep mutants 1125:1131 arg1 Y316F 1152:1156 Y316F 1152:1156 Y316F 1152:1156 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 56 dep mutants 1125:1131 arg1 W261Y 1134:1138 W261Y 1134:1138 W261Y 1134:1138 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 56 dep mutants 1125:1131 arg1 mutants 1125:1131 three positive mutants 1110:1131 three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood 1110:1191 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 3 57 theme Alternaria 401:410 arg1 sp 412:413 Alternaria sp 401:413 Alternaria sp 401:413 Our previous work reported that a wild-type α-L-rhamnosidase (RhaL1) from Alternaria sp. 27352363 9 58 theme site 1700:1703 arg1 cavity 1705:1710 the active site cavity 1689:1710 the active site cavity 1689:1710 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 6 59 theme hydrogen 877:884 arg1 bonds 886:890 direct or solvent-mediated hydrogen bonds 850:890 direct or solvent-mediated hydrogen bonds 850:890 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 7 60 theme prominent 1287:1295 arg1 increase 1297:1304 a prominent increase 1285:1304 a prominent increase (43.7 %) in relative yield 1285:1331 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 7 60 theme prominent 1287:1295 arg1 % 1312:1312 43.7 % 1307:1312 43.7 % 1307:1312 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 6 61 theme designed 782:789 arg1 mutations 791:799 rationally designed mutations 771:799 rationally designed mutations 771:799 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 62 theme RhaL1 1085:1089 arg1 active-site 1070:1080 active-site 1070:1080 active-site of RhaL1 1070:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 62 theme RhaL1 1085:1089 arg1 RhaL1 1085:1089 RhaL1 1085:1089 RhaL1 1085:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 9 63 theme enzyme 1753:1758 arg1 behavior 1737:1744 the catalysis behavior 1723:1744 the catalysis behavior of the enzyme 1723:1758 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 5 64 theme site-directed 693:705 arg1 mutagenesis 707:717 site-directed mutagenesis 693:717 site-directed mutagenesis of 11 residues 693:732 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 6 65 theme solvent-mediated 860:875 arg1 bonds 886:890 direct or solvent-mediated hydrogen bonds 850:890 direct or solvent-mediated hydrogen bonds 850:890 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 5 66 theme essential 646:654 arg1 residues 637:644 the amino acid residues 622:644 the amino acid residues essential for the catalytic activity of RhaL1 622:690 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 8 67 theme residue 1536:1542 arg1 position 1496:1503 the spatial position 1484:1503 the spatial position of the putative catalytic acid residue Asp257 1484:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 6 68 theme amino 815:819 arg1 residues 826:833 the critical amino acid residues 802:833 the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 802:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 5 69 theme reverse 577:583 arg1 reaction 596:603 reverse hydrolysis reaction 577:603 reverse hydrolysis reaction 577:603 To enhance the yield of reverse hydrolysis reaction and to determine the amino acid residues essential for the catalytic activity of RhaL1, site-directed mutagenesis of 11 residues was performed in this study. 27352363 1 70 theme rational 127:134 arg1 design 136:141 rational design 127:141 rational design 127:141 L1 to enhance synthesis yield of reverse hydrolysis based on rational design. 27352363 2 71 from applications 276:287 arg1 variety 294:300 a variety 292:300 a variety of industrial processes 292:324 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 2 71 from applications 276:287 arg1 processes 316:324 industrial processes 305:324 industrial processes 305:324 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 9 72 theme hydrolysis 1618:1627 arg1 reaction 1629:1636 hydrolysis reaction 1618:1636 hydrolysis reaction 1618:1636 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 6 73 theme direct 850:855 arg1 bonds 886:890 direct or solvent-mediated hydrogen bonds 850:890 direct or solvent-mediated hydrogen bonds 850:890 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 9 74 theme polar 1672:1676 arg1 residue 1678:1684 a polar residue 1670:1684 a polar residue in the active site cavity 1670:1710 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 6 75 from donor 1024:1028 arg1 active-site 1070:1080 active-site 1070:1080 active-site of RhaL1 1070:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 6 75 from donor 1024:1028 arg1 RhaL1 1085:1089 RhaL1 1085:1089 RhaL1 1085:1089 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 2 76 theme rhamnose 201:208 arg1 release 190:196 the hydrolytic release 175:196 the hydrolytic release of rhamnose from polysaccharides and glycosides 175:244 The α-L-rhamnosidase catalyzes the hydrolytic release of rhamnose from polysaccharides and glycosides and is widely used due to its applications in a variety of industrial processes. 27352363 7 77 theme reverse 1261:1267 arg1 hydrolysis 1269:1278 the reverse hydrolysis 1257:1278 the reverse hydrolysis 1257:1278 From the three positive variants, mutant W261Y accelerated the reverse hydrolysis with a prominent increase (43.7 %) in relative yield compared to the wild-type enzyme. 27352363 8 78 theme acid 1531:1534 arg1 residue 1536:1542 the putative catalytic acid residue Asp257 1508:1549 the putative catalytic acid residue Asp257 1508:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27352363 6 79 theme hydrophobic 990:1000 arg1 pocket 1002:1007 the hydrophobic pocket 986:1007 the hydrophobic pocket 986:1007 Through rationally designed mutations, the critical amino acid residues which may form direct or solvent-mediated hydrogen bonds with donor rhamnose (Asp252, Asp257, Asp264, Glu530, Arg548, His553, and Trp555) and may form the hydrophobic pocket in stabilizing donor (Trp261, Tyr302, Tyr316, and Trp369) in active-site of RhaL1 were analyzed, and three positive mutants (W261Y, Y302F, and Y316F) with improved product yield stood out. 27352363 9 80 theme active 1693:1698 arg1 cavity 1705:1710 the active site cavity 1689:1710 the active site cavity 1689:1710 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 9 81 from profile 1607:1613 arg1 reaction 1629:1636 hydrolysis reaction 1618:1636 hydrolysis reaction 1618:1636 Mutant W261Y also exhibited a shift in the pH-activity profile in hydrolysis reaction, indicating that introducing of a polar residue in the active site cavity may affect the catalysis behavior of the enzyme. 27352363 4 82 with reaction 499:506 arg1 rhamnose 525:532 inexpensive rhamnose 513:532 inexpensive rhamnose as glycosyl donor 513:550 L1 could synthesize rhamnose-containing chemicals (RCCs) though reverse hydrolysis reaction with inexpensive rhamnose as glycosyl donor. 27352363 8 83 theme spatial 1488:1494 arg1 position 1496:1503 the spatial position 1484:1503 the spatial position of the putative catalytic acid residue Asp257 1484:1549 Based on the 3D structural modeling, we supposed that the improved yield of mutant W261Y is due to the adjustment of the spatial position of the putative catalytic acid residue Asp257. 27012207 0 0 theme neuraminidase 75:87 arg1 fucose 7:12 Core-6 fucose 0:12 Core-6 fucose 0:12 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 0 0 theme neuraminidase 75:87 arg1 oligomerization 22:36 the oligomerization 18:36 the oligomerization of the 1918 pandemic influenza viral neuraminidase 18:87 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 5 1 from form 668:671 arg1 glycans 644:650 the glycans 640:650 the glycans from the active form 640:671 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 3 2 theme form 418:421 arg1 discovery 395:403 the discovery 391:403 the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation 391:519 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 0 3 theme viral 69:73 arg1 neuraminidase 75:87 the 1918 pandemic influenza viral neuraminidase 41:87 the 1918 pandemic influenza viral neuraminidase 41:87 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 7 4 theme azido 977:981 arg1 fucose 983:988 azido fucose 977:988 azido fucose 977:988 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 27012207 5 5 theme 82 746:747 arg1 % 748:748 % 748:748 % 748:748 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 5 6 theme % 748:748 arg1 fucosylated 757:767 82% core-6 fucosylated 746:767 82% core-6 fucosylated 746:767 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 4 7 from forms 562:566 arg1 N-glycans 542:550 the N-glycans 538:550 the N-glycans from both forms 538:566 In this report, the N-glycans from both forms were released and characterized by mass spectrometry. 27012207 7 8 theme in vitro 951:958 arg1 incorporation 960:972 in vitro incorporation 951:972 in vitro incorporation of azido fucose and (3)H-labeled fucose 951:1012 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 27012207 3 9 theme inactive 441:448 arg1 form 450:453 an inactive form 438:453 an inactive form (dimer and monomer) of the protein due to different glycosylation 438:519 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 6 10 theme form 825:828 arg1 region 804:809 the stalk region 794:809 the stalk region of the active form 794:828 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 6 10 theme form 825:828 arg1 devoid 852:857 devoid 852:857 devoid 852:857 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 5 11 from form 737:740 arg1 glycans 711:717 the glycans 707:717 the glycans from the inactive form 707:740 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 3 12 theme form 450:453 arg1 discovery 395:403 the discovery 391:403 the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation 391:519 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 6 13 theme active 818:823 arg1 form 825:828 the active form 814:828 the active form 814:828 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 5 14 theme core-6 750:755 arg1 fucosylated 757:767 82% core-6 fucosylated 746:767 82% core-6 fucosylated 746:767 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 8 15 theme enzymatic 1113:1121 arg1 activity 1123:1130 the enzymatic activity 1109:1130 the enzymatic activity of the active form 1109:1149 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 6 16 link core-6-linked 862:874 arg1 fucose 876:881 core-6-linked fucose 862:881 core-6-linked fucose 862:881 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 1 17 theme pandemics 163:171 arg1 pandemics 163:171 the most deadly pandemics 147:171 the most deadly pandemics in human history 147:188 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 1 17 theme pandemics 163:171 arg1 one 140:142 one 140:142 one 140:142 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 1 18 theme most 151:154 arg1 pandemics 163:171 the most deadly pandemics 147:171 the most deadly pandemics in human history 147:188 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 5 19 theme 26 677:678 arg1 % 679:679 % 679:679 % 679:679 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 1 20 theme deadly 156:161 arg1 pandemics 163:171 the most deadly pandemics 147:171 the most deadly pandemics in human history 147:188 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 7 21 theme fucose 1007:1012 arg1 incorporation 960:972 in vitro incorporation 951:972 in vitro incorporation of azido fucose and (3)H-labeled fucose 951:1012 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 27012207 3 22 theme virus 357:361 arg1 antigen 342:348 the neuraminidase (NA) antigen 319:348 the neuraminidase (NA) antigen of the virus 319:361 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 0 23 theme Core-6 0:5 arg1 fucose 7:12 Core-6 fucose 0:12 Core-6 fucose 0:12 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 5 24 theme inactive 728:735 arg1 form 737:740 the inactive form 724:740 the inactive form 724:740 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 5 25 theme % 679:679 arg1 fucosylated 688:698 26% core-6 fucosylated 677:698 26% core-6 fucosylated 677:698 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 9 26 dep oligomerization 1282:1296 arg1 the 1278:1280 the 1278:1280 the 1278:1280 It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. 27012207 5 27 theme active 661:666 arg1 form 668:671 the active form 657:671 the active form 657:671 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 8 28 theme enzymatic 1212:1220 arg1 activity 1222:1229 the enzymatic activity 1208:1229 the enzymatic activity 1208:1229 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 1 29 theme human 176:180 arg1 history 182:188 human history 176:188 human history 176:188 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 3 30 theme neuraminidase 323:335 arg1 antigen 342:348 the neuraminidase (NA) antigen 319:348 the neuraminidase (NA) antigen of the virus 319:361 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 1 31 theme H1N1 99:102 arg1 responsible 124:134 responsible 124:134 responsible 124:134 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 1 31 theme H1N1 99:102 arg1 virus 114:118 The 1918 H1N1 influenza virus 90:118 The 1918 H1N1 influenza virus 90:118 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 6 32 theme stalk 798:802 arg1 region 804:809 the stalk region 794:809 the stalk region of the active form 794:828 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 6 32 theme stalk 798:802 arg1 devoid 852:857 devoid 852:857 devoid 852:857 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 9 33 theme subsequent 1302:1311 arg1 activation 1313:1322 subsequent activation 1302:1322 subsequent activation 1302:1322 It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. 27012207 7 34 dep fucose 1007:1012 arg1 3 995:995 3 995:995 3 995:995 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 27012207 7 35 theme fucose 983:988 arg1 incorporation 960:972 in vitro incorporation 951:972 in vitro incorporation of azido fucose and (3)H-labeled fucose 951:1012 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 27012207 9 36 theme core-6 1254:1259 arg1 fucose 1261:1266 core-6 fucose 1254:1266 core-6 fucose 1254:1266 It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. 27012207 8 37 theme active 1139:1144 arg1 form 1146:1149 the active form 1135:1149 the active form 1135:1149 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 3 38 theme protein 482:488 arg1 form 450:453 an inactive form 438:453 an inactive form (dimer and monomer) of the protein due to different glycosylation 438:519 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 3 38 theme protein 482:488 arg1 tetramer 424:431 tetramer 424:431 tetramer 424:431 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 3 38 theme protein 482:488 arg1 form 418:421 an active form 408:421 an active form (tetramer) 408:432 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 6 39 theme fucose 876:881 arg1 devoid 852:857 devoid 852:857 devoid 852:857 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 6 39 theme fucose 876:881 arg1 region 804:809 the stalk region 794:809 the stalk region of the active form 794:828 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 0 40 theme 1918 45:48 arg1 pandemic 50:57 1918 pandemic 45:57 the 1918 pandemic influenza viral neuraminidase 41:87 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 5 41 theme core-6 681:686 arg1 fucosylated 688:698 26% core-6 fucosylated 677:698 26% core-6 fucosylated 677:698 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 5 42 contain had 673:675 arg2 fucosylated 688:698 26% core-6 fucosylated 677:698 26% core-6 fucosylated 677:698 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 5 42 contain had 673:675 arg1 glycans 644:650 the glycans 640:650 the glycans from the active form 640:671 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 1 43 theme influenza 104:112 arg1 responsible 124:134 responsible 124:134 responsible 124:134 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 1 43 theme influenza 104:112 arg1 virus 114:118 The 1918 H1N1 influenza virus 90:118 The 1918 H1N1 influenza virus 90:118 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 3 44 theme active 411:416 arg1 tetramer 424:431 tetramer 424:431 tetramer 424:431 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 3 44 theme active 411:416 arg1 form 418:421 an active form 408:421 an active form (tetramer) 408:432 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 3 45 theme different 497:505 arg1 glycosylation 507:519 different glycosylation 497:519 different glycosylation 497:519 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 8 46 theme fucose 1087:1092 arg1 incorporation 1070:1082 the incorporation 1066:1082 the incorporation of fucose 1066:1092 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 4 47 theme mass 603:606 arg1 spectrometry 608:619 mass spectrometry 603:619 mass spectrometry 603:619 In this report, the N-glycans from both forms were released and characterized by mass spectrometry. 27012207 6 48 theme core-6-linked 862:874 arg1 fucose 876:881 core-6-linked fucose 862:881 core-6-linked fucose 862:881 Even more surprisingly, the stalk region of the active form was almost completely devoid of core-6-linked fucose. 27012207 2 49 theme responsible 228:238 arg1 component 218:226 the structure component 204:226 the structure component responsible for its virulence 204:256 Yet to date, the structure component responsible for its virulence is still a mystery. 27012207 2 49 theme responsible 228:238 arg1 mystery 269:275 a mystery 267:275 a mystery 267:275 Yet to date, the structure component responsible for its virulence is still a mystery. 27012207 1 50 from pandemics 163:171 arg1 history 182:188 human history 176:188 human history 176:188 The 1918 H1N1 influenza virus was responsible for one of the most deadly pandemics in human history. 27012207 3 51 dep form 450:453 arg1 dimer 456:460 dimer 456:460 dimer 456:460 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 3 51 dep form 450:453 arg1 monomer 466:472 monomer 466:472 monomer 466:472 In order to search for such a component, the neuraminidase (NA) antigen of the virus was expressed, which led to the discovery of an active form (tetramer) and an inactive form (dimer and monomer) of the protein due to different glycosylation. 27012207 9 52 theme enzyme 1331:1336 arg1 oligomerization 1282:1296 oligomerization 1282:1296 oligomerization 1282:1296 It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. 27012207 9 52 theme enzyme 1331:1336 arg1 activation 1313:1322 subsequent activation 1302:1322 subsequent activation 1302:1322 It is postulated that core-6 fucose prohibits the oligomerization and subsequent activation of the enzyme. 27012207 0 53 theme influenza 59:67 arg1 neuraminidase 75:87 the 1918 pandemic influenza viral neuraminidase 41:87 the 1918 pandemic influenza viral neuraminidase 41:87 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 8 54 theme core-6 1166:1171 arg1 fucose 1173:1178 core-6 fucose 1166:1178 core-6 fucose 1166:1178 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 8 55 theme form 1146:1149 arg1 activity 1123:1130 the enzymatic activity 1109:1130 the enzymatic activity of the active form 1109:1149 In addition, the incorporation of fucose did not change the enzymatic activity of the active form, implying that core-6 fucose is not directly involved in the enzymatic activity. 27012207 0 56 theme pandemic 50:57 arg1 neuraminidase 75:87 the 1918 pandemic influenza viral neuraminidase 41:87 the 1918 pandemic influenza viral neuraminidase 41:87 Core-6 fucose and the oligomerization of the 1918 pandemic influenza viral neuraminidase. 27012207 2 57 theme structure 208:216 arg1 component 218:226 the structure component 204:226 the structure component responsible for its virulence 204:256 Yet to date, the structure component responsible for its virulence is still a mystery. 27012207 2 57 theme structure 208:216 arg1 mystery 269:275 a mystery 267:275 a mystery 267:275 Yet to date, the structure component responsible for its virulence is still a mystery. 27012207 5 58 contain had 742:744 arg2 fucosylated 757:767 82% core-6 fucosylated 746:767 82% core-6 fucosylated 746:767 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 5 58 contain had 742:744 arg1 glycans 711:717 the glycans 707:717 the glycans from the inactive form 707:740 It was found that the glycans from the active form had 26% core-6 fucosylated, while the glycans from the inactive form had 82% core-6 fucosylated. 27012207 7 59 theme H-labeled 997:1005 arg1 fucose 1007:1012 (3)H-labeled fucose 994:1012 (3)H-labeled fucose 994:1012 These findings were further supported by the results obtained from in vitro incorporation of azido fucose and (3)H-labeled fucose using core-6 fucosyltransferase, FUT8. 26117288 5 0 from production 779:788 arg1 LSCs 797:800 the LSCs 793:800 the LSCs 793:800 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 7 1 theme N-glycan 1111:1118 arg1 processing 1120:1129 N-glycan processing 1111:1129 N-glycan processing 1111:1129 These results suggest that SW inhibits N-glycan processing, which attenuates LH receptor activity, which, in turn, reduces progesterone production by CL. 26117288 1 2 theme potent 141:146 arg1 swainsonine 121:131 The indolizidine alkaloid swainsonine 95:131 The indolizidine alkaloid swainsonine (SW) 95:136 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 2 theme potent 141:146 arg1 inhibitor 148:156 a potent inhibitor 139:156 a potent inhibitor of Golgi α-mannosidase II 139:182 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 6 3 theme steroidogenic 886:898 arg1 protein 917:923 steroidogenic acute regulatory protein 886:923 steroidogenic acute regulatory protein 886:923 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 4 4 theme inhibition 578:587 arg1 effect 548:553 the effect 544:553 the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs 544:658 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 0 5 theme luteal 81:86 arg1 cells 88:92 bovine luteal cells 74:92 bovine luteal cells 74:92 Alteration of glycan structures by swainsonine affects steroidogenesis in bovine luteal cells. 26117288 4 6 theme present 513:519 arg1 study 521:525 the present study 509:525 the present study 509:525 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 1 7 theme toxicity 241:248 arg1 syndrome 250:257 toxicity syndrome 241:257 toxicity syndrome known as locoism in livestock 241:287 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 7 8 theme receptor 1152:1159 arg1 activity 1161:1168 LH receptor activity 1149:1168 LH receptor activity 1149:1168 These results suggest that SW inhibits N-glycan processing, which attenuates LH receptor activity, which, in turn, reduces progesterone production by CL. 26117288 5 9 contain had 735:737 arg1 it 732:733 it 732:733 it 732:733 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 5 9 contain had 735:737 arg2 effect 742:747 no effect 739:747 no effect 739:747 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 6 10 theme progesterone 843:854 arg1 production 856:865 progesterone production 843:865 progesterone production 843:865 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 0 11 from steroidogenesis 55:69 arg1 cells 88:92 bovine luteal cells 74:92 bovine luteal cells 74:92 Alteration of glycan structures by swainsonine affects steroidogenesis in bovine luteal cells. 26117288 2 12 theme serious 365:371 arg1 disorders 373:381 serious disorders 365:381 serious disorders in reproductive functions of livestock 365:420 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 5 13 from glycosylation 701:713 arg1 LSCs 722:725 the LSCs 718:725 the LSCs 718:725 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 6 14 theme protein 917:923 arg1 expressions 871:881 expressions 871:881 expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 871:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 14 theme protein 917:923 arg1 production 856:865 progesterone production 843:865 progesterone production 843:865 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 15 theme receptor 1053:1060 arg1 activity 1062:1069 LH receptor activity 1050:1069 LH receptor activity 1050:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 2 16 theme livestock 412:420 arg1 functions 399:407 reproductive functions 386:407 reproductive functions of livestock 386:420 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 6 17 theme LSCs 980:983 arg1 Pretreatment 964:975 Pretreatment 964:975 Pretreatment of LSCs with SW 964:991 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 18 with Pretreatment 964:975 arg1 SW 990:991 SW 990:991 SW 990:991 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 1 19 theme Golgi 161:165 arg1 α-mannosidase 167:179 Golgi α-mannosidase II 161:182 Golgi α-mannosidase II 161:182 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 7 20 theme progesterone 1195:1206 arg1 production 1208:1217 progesterone production 1195:1217 progesterone production by CL 1195:1223 These results suggest that SW inhibits N-glycan processing, which attenuates LH receptor activity, which, in turn, reduces progesterone production by CL. 26117288 1 21 theme α-mannosidase 167:179 arg1 swainsonine 121:131 The indolizidine alkaloid swainsonine 95:131 The indolizidine alkaloid swainsonine (SW) 95:136 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 21 theme α-mannosidase 167:179 arg1 inhibitor 148:156 a potent inhibitor 139:156 a potent inhibitor of Golgi α-mannosidase II 139:182 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 5 22 from viability 752:760 arg1 LSCs 797:800 the LSCs 793:800 the LSCs 793:800 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 7 23 theme LH 1149:1150 arg1 activity 1161:1168 LH receptor activity 1149:1168 LH receptor activity 1149:1168 These results suggest that SW inhibits N-glycan processing, which attenuates LH receptor activity, which, in turn, reduces progesterone production by CL. 26117288 1 24 from locoism 268:274 arg1 livestock 279:287 livestock 279:287 livestock 279:287 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 6 25 dep I. 961:962 arg1 suppressed 993:1002 suppressed 993:1002 suppressed 993:1002 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 2 26 theme reproductive 386:397 arg1 functions 399:407 reproductive functions 386:407 reproductive functions of livestock 386:420 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 0 27 theme structures 21:30 arg1 Alteration 0:9 Alteration 0:9 Alteration of glycan structures by swainsonine 0:45 Alteration of glycan structures by swainsonine affects steroidogenesis in bovine luteal cells. 26117288 2 28 from disorders 373:381 arg1 functions 399:407 reproductive functions 386:407 reproductive functions of livestock 386:420 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 2 29 theme Locoweed 290:297 arg1 consumption 299:309 Locoweed consumption 290:309 Locoweed consumption 290:309 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 0 30 theme glycan 14:19 arg1 structures 21:30 glycan structures 14:30 glycan structures 14:30 Alteration of glycan structures by swainsonine affects steroidogenesis in bovine luteal cells. 26117288 4 31 theme N-glycan 558:565 arg1 inhibition 578:587 N-glycan processing inhibition 558:587 N-glycan processing inhibition by SW 558:593 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 1 32 theme alkaloid 112:119 arg1 SW 134:135 SW 134:135 SW 134:135 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 32 theme alkaloid 112:119 arg1 swainsonine 121:131 The indolizidine alkaloid swainsonine 95:131 The indolizidine alkaloid swainsonine (SW) 95:136 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 32 theme alkaloid 112:119 arg1 inhibitor 148:156 a potent inhibitor 139:156 a potent inhibitor of Golgi α-mannosidase II 139:182 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 4 33 theme processing 567:576 arg1 inhibition 578:587 N-glycan processing inhibition 558:587 N-glycan processing inhibition by SW 558:593 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 6 34 dep protein 917:923 arg1 class 955:959 B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 953:1069 B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 953:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 4 35 theme bovine 648:653 arg1 LSCs 655:658 cultured bovine LSCs 639:658 cultured bovine LSCs 639:658 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 4 36 from viability 626:634 arg1 LSCs 655:658 cultured bovine LSCs 639:658 cultured bovine LSCs 639:658 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 3 37 theme luteal 456:461 arg1 LSCs 484:487 LSCs 484:487 LSCs 484:487 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 3 37 theme luteal 456:461 arg1 cells 477:481 luteal steroidogenic cells 456:481 luteal steroidogenic cells (LSCs) 456:488 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 3 38 theme SW 450:451 arg1 effects 439:445 direct effects 432:445 direct effects of SW on luteal steroidogenic cells (LSCs) 432:488 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 6 39 theme B 953:953 arg1 class 955:959 B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 953:1069 B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 953:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 3 40 theme direct 432:437 arg1 effects 439:445 direct effects 432:445 direct effects of SW on luteal steroidogenic cells (LSCs) 432:488 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 4 41 from effect 548:553 arg1 viability 626:634 viability 626:634 viability 626:634 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 4 41 from effect 548:553 arg1 production 611:620 progesterone production 598:620 progesterone production 598:620 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 3 42 theme steroidogenic 463:475 arg1 LSCs 484:487 LSCs 484:487 LSCs 484:487 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 3 42 theme steroidogenic 463:475 arg1 cells 477:481 luteal steroidogenic cells 456:481 luteal steroidogenic cells (LSCs) 456:488 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 5 43 theme progesterone 766:777 arg1 production 779:788 progesterone production 766:788 progesterone production 766:788 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 6 44 theme type 948:951 arg1 expressions 871:881 expressions 871:881 expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity 871:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 44 theme type 948:951 arg1 production 856:865 progesterone production 843:865 progesterone production 843:865 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 5 45 gly glycosylation 701:713 arg1 LSCs 722:725 the LSCs 718:725 the LSCs 718:725 Swainsonine (0.1 μg/mL) fully inhibited glycosylation in the LSCs, but it had no effect on viability and progesterone production in the LSCs. 26117288 6 46 theme acute 900:904 arg1 protein 917:923 steroidogenic acute regulatory protein 886:923 steroidogenic acute regulatory protein 886:923 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 47 theme receptor 939:946 arg1 type 948:951 scavenger receptor type 929:951 scavenger receptor type 929:951 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 48 theme regulatory 906:915 arg1 protein 917:923 steroidogenic acute regulatory protein 886:923 steroidogenic acute regulatory protein 886:923 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 49 theme scavenger 929:937 arg1 type 948:951 scavenger receptor type 929:951 scavenger receptor type 929:951 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 6 50 dep class 955:959 arg1 I. 961:962 I. 961:962 I. 961:962 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 2 51 theme progesterone 327:338 arg1 concentration 340:352 serum progesterone concentration 321:352 serum progesterone concentration 321:352 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 1 52 theme indolizidine 99:110 arg1 SW 134:135 SW 134:135 SW 134:135 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 52 theme indolizidine 99:110 arg1 swainsonine 121:131 The indolizidine alkaloid swainsonine 95:131 The indolizidine alkaloid swainsonine (SW) 95:136 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 52 theme indolizidine 99:110 arg1 inhibitor 148:156 a potent inhibitor 139:156 a potent inhibitor of Golgi α-mannosidase II 139:182 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 53 theme primary 203:209 arg1 toxicant 211:218 the primary toxicant 199:218 the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock 199:287 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 1 54 from toxicant 211:218 arg1 locoweeds 223:231 locoweeds 223:231 locoweeds causing toxicity syndrome known as locoism in livestock 223:287 The indolizidine alkaloid swainsonine (SW), a potent inhibitor of Golgi α-mannosidase II, is considered the primary toxicant in locoweeds causing toxicity syndrome known as locoism in livestock. 26117288 6 55 theme LH 1050:1051 arg1 receptor 1053:1060 LH receptor 1050:1060 LH receptor activity 1050:1069 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 3 56 from effects 439:445 arg1 LSCs 484:487 LSCs 484:487 LSCs 484:487 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 3 56 from effects 439:445 arg1 cells 477:481 luteal steroidogenic cells 456:481 luteal steroidogenic cells (LSCs) 456:488 However, direct effects of SW on luteal steroidogenic cells (LSCs) remain unclear. 26117288 2 57 theme serum 321:325 arg1 concentration 340:352 serum progesterone concentration 321:352 serum progesterone concentration 321:352 Locoweed consumption decreases serum progesterone concentration and causes serious disorders in reproductive functions of livestock. 26117288 0 58 theme bovine 74:79 arg1 cells 88:92 bovine luteal cells 74:92 bovine luteal cells 74:92 Alteration of glycan structures by swainsonine affects steroidogenesis in bovine luteal cells. 26117288 4 59 theme cultured 639:646 arg1 LSCs 655:658 cultured bovine LSCs 639:658 cultured bovine LSCs 639:658 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 6 60 theme Luteinizing 803:813 arg1 hormone 815:821 Luteinizing hormone 803:821 Luteinizing hormone 803:821 Luteinizing hormone is known to promote progesterone production and expressions of steroidogenic acute regulatory protein and scavenger receptor type B class I. Pretreatment of LSCs with SW suppressed each of these effects, possibly by inhibiting LH receptor activity. 26117288 4 61 from production 611:620 arg1 LSCs 655:658 cultured bovine LSCs 639:658 cultured bovine LSCs 639:658 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26117288 4 62 theme progesterone 598:609 arg1 production 611:620 progesterone production 598:620 progesterone production 598:620 In the present study, we investigated the effect of N-glycan processing inhibition by SW on progesterone production and viability in cultured bovine LSCs. 26492619 3 0 from O-glycan 389:396 arg1 Thr52 401:405 Thr52 401:405 Thr52 of PLAG3 401:414 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 8 1 theme LpMab-10 992:999 arg1 recognition 1001:1011 LpMab-10 recognition 992:1011 LpMab-10 recognition 992:1011 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 8 2 theme PLAG1 970:974 arg1 Thr34 961:965 The Thr34 957:965 The Thr34 of PLAG1 957:974 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 8 2 theme PLAG1 970:974 arg1 critical 979:986 critical 979:986 critical 979:986 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 1 3 theme transmembrane 155:167 arg1 Podoplanin 125:134 Podoplanin 125:134 Podoplanin (PDPN) 125:141 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 1 3 theme transmembrane 155:167 arg1 sialoglycoprotein 169:185 a type-I transmembrane sialoglycoprotein 146:185 a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus 146:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 5 4 theme PLAG1/2 734:740 arg1 domain 742:747 PLAG1/2 domain 734:747 PLAG1/2 domain 734:747 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 6 5 theme cancer 786:791 arg1 cells 793:797 cancer cells 786:797 cancer cells 786:797 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 2 6 dep repeats 304:310 arg1 PLAG2 341:345 PLAG2 341:345 PLAG2 341:345 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 6 dep repeats 304:310 arg1 PLAG3 352:356 PLAG3 352:356 PLAG3 352:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 6 dep repeats 304:310 arg1 PLAG1 334:338 PLAG1 334:338 PLAG1 334:338 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 6 dep repeats 304:310 arg1 repeats 304:310 three tandem repeats 291:310 three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3 291:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 10 7 theme monoclonal 1239:1248 arg1 antibodies 1250:1259 monoclonal antibodies 1239:1259 monoclonal antibodies 1239:1259 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 10 7 theme monoclonal 1239:1248 arg1 epitopes 1298:1305 epitopes 1298:1305 epitopes of PDPN 1298:1313 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 0 8 theme Podoplanin 113:122 arg1 Thr34 98:102 Thr34 98:102 Thr34 of Human Podoplanin 98:122 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 7 9 theme flow 941:944 arg1 cytometry 946:954 flow cytometry 941:954 flow cytometry 941:954 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 7 10 theme minimum 852:858 arg1 epitope 860:866 The minimum epitope 848:866 The minimum epitope of LpMab-10 848:878 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 7 10 theme minimum 852:858 arg1 Glu33-Gly45 898:908 Glu33-Gly45 898:908 Glu33-Gly45 of PDPN 898:916 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 0 11 theme Human 107:111 arg1 Podoplanin 113:122 Human Podoplanin 107:122 Human Podoplanin 107:122 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 7 12 theme Western 924:930 arg1 blot 932:935 Western blot 924:935 Western blot 924:935 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 4 13 theme human 596:600 arg1 PDPN 602:605 human PDPN 596:605 human PDPN 596:605 In contrast, the glycosylation of Thr34 of PLAG1 of human PDPN remains to be clarified. 26492619 2 14 theme tandem 297:302 arg1 repeats 304:310 three tandem repeats 291:310 three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3 291:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 14 theme tandem 297:302 arg1 PLAG3 352:356 PLAG3 352:356 PLAG3 352:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 14 theme tandem 297:302 arg1 PLAG1 334:338 PLAG1 334:338 PLAG1 334:338 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 14 theme tandem 297:302 arg1 PLAG2 341:345 PLAG2 341:345 PLAG2 341:345 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 7 15 theme PDPN 913:916 arg1 epitope 860:866 The minimum epitope 848:866 The minimum epitope of LpMab-10 848:878 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 7 15 theme PDPN 913:916 arg1 Glu33-Gly45 898:908 Glu33-Gly45 898:908 Glu33-Gly45 of PDPN 898:916 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 3 16 theme platelet-aggregating 505:524 arg1 activity 526:533 platelet-aggregating activity 505:533 platelet-aggregating activity of PDPN 505:541 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 3 17 theme PLAG 375:378 arg1 domains 380:386 the three PLAG domains 365:386 the three PLAG domains 365:386 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 10 18 theme different 1269:1277 arg1 Ser/Thr 1279:1285 different Ser/Thr 1269:1285 different Ser/Thr 1269:1285 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 4 19 gly glycosylation 561:573 arg1 Thr34 578:582 Thr34 578:582 Thr34 of PLAG1 of human PDPN 578:605 In contrast, the glycosylation of Thr34 of PLAG1 of human PDPN remains to be clarified. 26492619 2 20 theme acids 327:331 arg1 repeats 304:310 three tandem repeats 291:310 three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3 291:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 20 theme acids 327:331 arg1 PLAG3 352:356 PLAG3 352:356 PLAG3 352:356 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 20 theme acids 327:331 arg1 PLAG1 334:338 PLAG1 334:338 PLAG1 334:338 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 2 20 theme acids 327:331 arg1 PLAG2 341:345 PLAG2 341:345 PLAG2 341:345 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 1 21 theme platelet 204:211 arg1 domain 244:249 a platelet aggregation-stimulating (PLAG) domain 202:249 a platelet aggregation-stimulating (PLAG) domain in the N-terminus 202:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 0 22 theme Antibody 26:33 arg1 Development 0:10 Development 0:10 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain 0:86 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 0 23 gly Non-glycosylated 56:71 arg1 Domain 81:86 Non-glycosylated PLAG1/2 Domain 56:86 Non-glycosylated PLAG1/2 Domain 56:86 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 6 24 theme endogenous 767:776 arg1 PDPN 778:781 endogenous PDPN 767:781 endogenous PDPN of cancer cells and normal cells independently of glycosylation 767:845 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 3 25 theme PDPN 538:541 arg1 activity 526:533 platelet-aggregating activity 505:533 platelet-aggregating activity of PDPN 505:541 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 10 26 theme PDPN 1310:1313 arg1 epitopes 1298:1305 epitopes 1298:1305 epitopes of PDPN 1298:1313 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 1 27 theme aggregation-stimulating 213:235 arg1 domain 244:249 a platelet aggregation-stimulating (PLAG) domain 202:249 a platelet aggregation-stimulating (PLAG) domain in the N-terminus 202:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 0 28 theme Monoclonal 15:24 arg1 Antibody 26:33 Monoclonal Antibody LpMab-10 15:42 Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain 15:86 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 9 29 theme tumor 1217:1221 arg1 cells 1223:1227 PDPN-expressing tumor cells 1201:1227 PDPN-expressing tumor cells 1201:1227 In immunocytochemical and immunohistochemical analyses, LpMab-10 strongly detected PDPN-expressing tumor cells. 26492619 3 30 theme C-type 445:450 arg1 CLEC-2 476:481 CLEC-2 476:481 CLEC-2 476:481 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 3 30 theme C-type 445:450 arg1 receptor-2 464:473 C-type lectin-like receptor-2 445:473 C-type lectin-like receptor-2 (CLEC-2) 445:482 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 5 31 theme novel 673:677 arg1 LpMab-10 710:717 LpMab-10 710:717 LpMab-10 710:717 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 5 31 theme novel 673:677 arg1 antibody 700:707 a novel anti-PDPN monoclonal antibody 671:707 a novel anti-PDPN monoclonal antibody 671:707 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 6 32 theme glycosylation 833:845 arg1 PDPN 778:781 endogenous PDPN 767:781 endogenous PDPN of cancer cells and normal cells independently of glycosylation 767:845 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 3 33 theme PLAG3 410:414 arg1 Thr52 401:405 Thr52 401:405 Thr52 of PLAG3 401:414 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 1 34 theme PLAG 238:241 arg1 domain 244:249 a platelet aggregation-stimulating (PLAG) domain 202:249 a platelet aggregation-stimulating (PLAG) domain in the N-terminus 202:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 10 35 gly O-glycosylated 1386:1399 arg1 PDPN 1377:1380 PDPN 1377:1380 PDPN 1377:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 10 35 gly O-glycosylated 1386:1399 arg1 residues 1365:1372 Ser/Thr residues 1357:1372 Ser/Thr residues of PDPN 1357:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 10 36 theme Ser/Thr 1357:1363 arg1 PDPN 1377:1380 PDPN 1377:1380 PDPN 1377:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 10 36 theme Ser/Thr 1357:1363 arg1 residues 1365:1372 Ser/Thr residues 1357:1372 Ser/Thr residues of PDPN 1357:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 5 37 theme anti-PDPN 679:687 arg1 LpMab-10 710:717 LpMab-10 710:717 LpMab-10 710:717 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 5 37 theme anti-PDPN 679:687 arg1 antibody 700:707 a novel anti-PDPN monoclonal antibody 671:707 a novel anti-PDPN monoclonal antibody 671:707 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 1 38 contain possesses 192:200 arg2 domain 244:249 a platelet aggregation-stimulating (PLAG) domain 202:249 a platelet aggregation-stimulating (PLAG) domain in the N-terminus 202:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 1 38 contain possesses 192:200 arg1 Podoplanin 125:134 Podoplanin 125:134 Podoplanin (PDPN) 125:141 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 1 38 contain possesses 192:200 arg1 sialoglycoprotein 169:185 a type-I transmembrane sialoglycoprotein 146:185 a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus 146:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 0 39 theme PLAG1/2 73:79 arg1 Domain 81:86 Non-glycosylated PLAG1/2 Domain 56:86 Non-glycosylated PLAG1/2 Domain 56:86 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 5 40 theme monoclonal 689:698 arg1 LpMab-10 710:717 LpMab-10 710:717 LpMab-10 710:717 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 5 40 theme monoclonal 689:698 arg1 antibody 700:707 a novel anti-PDPN monoclonal antibody 671:707 a novel anti-PDPN monoclonal antibody 671:707 Herein, we developed and characterized a novel anti-PDPN monoclonal antibody, LpMab-10, which targets PLAG1/2 domain. 26492619 1 41 from domain 244:249 arg1 N-terminus 258:267 the N-terminus 254:267 the N-terminus 254:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 0 42 theme Non-glycosylated 56:71 arg1 Domain 81:86 Non-glycosylated PLAG1/2 Domain 56:86 Non-glycosylated PLAG1/2 Domain 56:86 Development of Monoclonal Antibody LpMab-10 Recognizing Non-glycosylated PLAG1/2 Domain Including Thr34 of Human Podoplanin. 26492619 6 43 theme cells 810:814 arg1 PDPN 778:781 endogenous PDPN 767:781 endogenous PDPN of cancer cells and normal cells independently of glycosylation 767:845 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 9 44 theme PDPN-expressing 1201:1215 arg1 cells 1223:1227 PDPN-expressing tumor cells 1201:1227 PDPN-expressing tumor cells 1201:1227 In immunocytochemical and immunohistochemical analyses, LpMab-10 strongly detected PDPN-expressing tumor cells. 26492619 1 45 gly sialoglycoprotein 169:185 arg1 Podoplanin 125:134 Podoplanin 125:134 Podoplanin (PDPN) 125:141 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 1 45 gly sialoglycoprotein 169:185 arg1 sialoglycoprotein 169:185 a type-I transmembrane sialoglycoprotein 146:185 a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus 146:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 6 46 theme normal 803:808 arg1 cells 810:814 normal cells 803:814 normal cells 803:814 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 9 47 theme immunohistochemical 1144:1162 arg1 analyses 1164:1171 immunohistochemical analyses 1144:1171 immunohistochemical analyses 1144:1171 In immunocytochemical and immunohistochemical analyses, LpMab-10 strongly detected PDPN-expressing tumor cells. 26492619 3 48 theme lectin-like 452:462 arg1 CLEC-2 476:481 CLEC-2 476:481 CLEC-2 476:481 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 3 48 theme lectin-like 452:462 arg1 receptor-2 464:473 C-type lectin-like receptor-2 445:473 C-type lectin-like receptor-2 (CLEC-2) 445:482 Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. 26492619 2 49 theme amino 321:325 arg1 acids 327:331 eight amino acids 315:331 eight amino acids 315:331 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 8 50 gly O-glycosylated 1102:1115 arg1 Thr34 1080:1084 Thr34 1080:1084 Thr34 of PLAG1 1080:1093 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 7 51 theme LpMab-10 871:878 arg1 epitope 860:866 The minimum epitope 848:866 The minimum epitope of LpMab-10 848:878 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 7 51 theme LpMab-10 871:878 arg1 Glu33-Gly45 898:908 Glu33-Gly45 898:908 Glu33-Gly45 of PDPN 898:916 The minimum epitope of LpMab-10 was identified as Glu33-Gly45 of PDPN using Western blot and flow cytometry. 26492619 4 52 theme Thr34 578:582 arg1 glycosylation 561:573 the glycosylation 557:573 the glycosylation of Thr34 of PLAG1 of human PDPN 557:605 In contrast, the glycosylation of Thr34 of PLAG1 of human PDPN remains to be clarified. 26492619 8 53 theme PLAG1 1089:1093 arg1 Thr34 1080:1084 Thr34 1080:1084 Thr34 of PLAG1 1080:1093 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 6 54 theme cells 793:797 arg1 PDPN 778:781 endogenous PDPN 767:781 endogenous PDPN of cancer cells and normal cells independently of glycosylation 767:845 LpMab-10 detects endogenous PDPN of cancer cells and normal cells independently of glycosylation. 26492619 2 55 theme PLAG 270:273 arg1 domain 275:280 PLAG domain 270:280 PLAG domain 270:280 PLAG domain includes three tandem repeats of eight amino acids: PLAG1, PLAG2, and PLAG3. 26492619 4 56 theme PDPN 602:605 arg1 PLAG1 587:591 PLAG1 587:591 PLAG1 of human PDPN 587:605 In contrast, the glycosylation of Thr34 of PLAG1 of human PDPN remains to be clarified. 26492619 10 57 theme PDPN 1377:1380 arg1 PDPN 1377:1380 PDPN 1377:1380 PDPN 1377:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 10 57 theme PDPN 1377:1380 arg1 residues 1365:1372 Ser/Thr residues 1357:1372 Ser/Thr residues of PDPN 1357:1380 By using monoclonal antibodies against different Ser/Thr, including epitopes of PDPN, it becomes possible to determine whether Ser/Thr residues of PDPN are O-glycosylated. 26492619 1 58 theme type-I 148:153 arg1 Podoplanin 125:134 Podoplanin 125:134 Podoplanin (PDPN) 125:141 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 1 58 theme type-I 148:153 arg1 sialoglycoprotein 169:185 a type-I transmembrane sialoglycoprotein 146:185 a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus 146:267 Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein that possesses a platelet aggregation-stimulating (PLAG) domain in the N-terminus. 26492619 8 59 theme LpMab-10 1046:1053 arg1 epitope 1055:1061 LpMab-10 epitope 1046:1061 LpMab-10 epitope 1046:1061 The Thr34 of PLAG1 is critical for LpMab-10 recognition, and O-glycan is not included in LpMab-10 epitope, indicating that Thr34 of PLAG1 is not O-glycosylated. 26492619 4 60 theme PLAG1 587:591 arg1 Thr34 578:582 Thr34 578:582 Thr34 of PLAG1 of human PDPN 578:605 In contrast, the glycosylation of Thr34 of PLAG1 of human PDPN remains to be clarified. 28170415 0 0 theme transgenic 85:94 arg1 cows 96:99 transgenic cows 85:99 transgenic cows 85:99 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 9 1 theme high 1208:1211 arg1 glycans 1221:1227 high mannose glycans 1208:1227 high mannose glycans (65%) 1208:1233 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 1 theme high 1208:1211 arg1 % 1232:1232 65% 1230:1232 65% 1230:1232 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 1 2 gly glycoproteins 157:169 arg1 glycoproteins 157:169 the most abundant bioactive glycoproteins 129:169 the most abundant bioactive glycoproteins in human milk 129:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 1 3 from glycoproteins 157:169 arg1 milk 180:183 human milk 174:183 human milk 174:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 6 4 gly sialylated 747:756 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 3 5 theme trace 313:317 arg1 amounts 319:325 trace amounts 313:325 trace amounts in bovine milk 313:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 9 6 theme complex/hybrid 1110:1123 arg1 N-glycans 1125:1133 mostly neutral complex/hybrid N-glycans 1095:1133 mostly neutral complex/hybrid N-glycans 1095:1133 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 5 7 theme Nano-LC-Chip-Q-TOF 621:638 arg1 Spectrometry 645:656 Nano-LC-Chip-Q-TOF Mass Spectrometry 621:656 Nano-LC-Chip-Q-TOF Mass Spectrometry 621:656 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 4 8 theme bovine 383:388 arg1 milk 390:393 bovine milk 383:393 bovine milk 383:393 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 1 9 theme abundant 138:145 arg1 glycoproteins 157:169 the most abundant bioactive glycoproteins 129:169 the most abundant bioactive glycoproteins in human milk 129:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 9 10 theme total 1155:1159 arg1 % 1138:1138 81% 1136:1138 81% 1136:1138 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 10 theme total 1155:1159 arg1 % 1146:1146 52% 1144:1146 52% of the total 1144:1159 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 10 theme total 1155:1159 arg1 total 1155:1159 total 1155:1159 total 1155:1159 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 11 11 theme more 1429:1432 arg1 similarities 1434:1445 more similarities 1429:1445 more similarities to bLF 1429:1452 Overall, this study suggests that rhLF N-glycans share more similarities to bLF than hLF. 28170415 5 12 theme Mass 640:643 arg1 Spectrometry 645:656 Nano-LC-Chip-Q-TOF Mass Spectrometry 621:656 Nano-LC-Chip-Q-TOF Mass Spectrometry 621:656 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 6 13 theme N-glycan 700:707 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 6 13 theme N-glycan 700:707 arg1 structures 709:718 N-glycan structures 700:718 N-glycan structures 700:718 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 6 14 gly fucosylated 731:741 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 0 15 theme cows 96:99 arg1 milk 77:80 the milk 73:80 the milk of transgenic cows 73:99 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 6 16 theme sialylated 747:756 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 3 17 from contrast 286:293 arg1 present 302:308 present 302:308 present 302:308 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 18 theme bovine 330:335 arg1 milk 337:340 bovine milk 330:340 bovine milk 330:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 4 19 theme human 458:462 arg1 hLF 477:479 hLF 477:479 hLF 477:479 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 4 19 theme human 458:462 arg1 lactoferrin 464:474 human lactoferrin 458:474 human lactoferrin (hLF) 458:480 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 6 20 theme additional 794:803 arg1 bioactivities 805:817 additional bioactivities 794:817 additional bioactivities 794:817 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 9 21 theme relative 1023:1030 arg1 abundances 1032:1041 the relative abundances 1019:1041 the relative abundances of N-glycan types 1019:1059 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 1 22 theme bioactive 147:155 arg1 glycoproteins 157:169 the most abundant bioactive glycoproteins 129:169 the most abundant bioactive glycoproteins in human milk 129:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 6 23 theme high 682:685 arg1 diversity 687:695 a high diversity 680:695 a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities 680:817 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 1 24 theme glycoproteins 157:169 arg1 one 122:124 one 122:124 one 122:124 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 1 24 theme glycoproteins 157:169 arg1 glycoproteins 157:169 the most abundant bioactive glycoproteins 129:169 the most abundant bioactive glycoproteins in human milk 129:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 8 25 with bLF 943:945 arg1 hLF 967:969 hLF 967:969 hLF 967:969 rhLF shared 16 N-glycan with bLF and 9 N-glycan with hLF while bLF shared 10 N-glycan with hLF. 28170415 5 26 theme human 600:604 arg1 lactoferrin 606:616 human lactoferrin 600:616 human lactoferrin 600:616 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 3 27 located present 302:308 arg1 amounts 319:325 trace amounts 313:325 trace amounts in bovine milk 313:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 27 located present 302:308 arg2 LF 296:297 LF 296:297 LF 296:297 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 27 located present 302:308 arg1 contrast 286:293 contrast 286:293 contrast 286:293 In contrast, LF is present in trace amounts in bovine milk. 28170415 9 28 theme mannose 1213:1219 arg1 glycans 1221:1227 high mannose glycans 1208:1227 high mannose glycans (65%) 1208:1233 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 28 theme mannose 1213:1219 arg1 % 1232:1232 65% 1230:1232 65% 1230:1232 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 3 29 attach present 302:308 arg1 amounts 319:325 trace amounts 313:325 trace amounts in bovine milk 313:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 29 attach present 302:308 arg2 LF 296:297 LF 296:297 LF 296:297 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 29 attach present 302:308 arg1 contrast 286:293 contrast 286:293 contrast 286:293 In contrast, LF is present in trace amounts in bovine milk. 28170415 11 30 theme rhLF 1408:1411 arg1 N-glycans 1413:1421 rhLF N-glycans 1408:1421 rhLF N-glycans 1408:1421 Overall, this study suggests that rhLF N-glycans share more similarities to bLF than hLF. 28170415 4 31 dep Efforts 343:349 arg1 increase 354:361 increase 354:361 to increase LF concentration in bovine milk 351:393 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 1 32 theme human 174:178 arg1 milk 180:183 human milk 174:183 human milk 174:183 Lactoferrin (LF) is one of the most abundant bioactive glycoproteins in human milk. 28170415 0 33 theme human 32:36 arg1 N-glycans 50:58 recombinant human lactoferrin N-glycans 20:58 recombinant human lactoferrin N-glycans 20:58 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 4 34 theme LF 363:364 arg1 concentration 366:378 LF concentration 363:378 LF concentration 363:378 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 7 35 from N-glycan 886:893 arg1 overall 905:911 overall 905:911 overall 905:911 rhLF, bLF and hLF had 23, 27 and 18 N-glycans respectively with 8 N-glycan in common overall. 28170415 5 36 theme bovine 571:576 arg1 lactoferrin 578:588 bovine lactoferrin 571:588 bovine lactoferrin (bLF) 571:594 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 36 theme bovine 571:576 arg1 bLF 591:593 bLF 591:593 bLF 591:593 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 0 37 theme recombinant 20:30 arg1 N-glycans 50:58 recombinant human lactoferrin N-glycans 20:58 recombinant human lactoferrin N-glycans 20:58 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 9 38 dep contain 1087:1093 arg1 % 1138:1138 81% 1136:1138 81% 1136:1138 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 38 dep contain 1087:1093 arg1 % 1146:1146 52% 1144:1146 52% of the total 1144:1159 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 38 dep contain 1087:1093 arg1 total 1155:1159 total 1155:1159 total 1155:1159 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 39 theme N-glycan 1046:1053 arg1 types 1055:1059 N-glycan types 1046:1059 N-glycan types 1046:1059 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 0 40 theme N-glycans 50:58 arg1 Characterization 0:15 Characterization 0:15 Characterization of recombinant human lactoferrin N-glycans 0:58 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 4 41 theme transgenic 431:440 arg1 cows 442:445 transgenic cows 431:445 transgenic cows 431:445 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 10 42 contain had 1326:1328 arg1 rhLF 1321:1324 rhLF 1321:1324 rhLF 1321:1324 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 10 42 contain had 1326:1328 arg1 bLF 1313:1315 bLF 1313:1315 bLF 1313:1315 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 10 42 contain had 1326:1328 arg2 % 1336:1336 only 9% 1330:1336 only 9% 1330:1336 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 10 42 contain had 1326:1328 arg2 % 1344:1344 20% 1342:1344 20% fucosylation 1342:1357 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 0 43 theme lactoferrin 38:48 arg1 N-glycans 50:58 recombinant human lactoferrin N-glycans 20:58 recombinant human lactoferrin N-glycans 20:58 Characterization of recombinant human lactoferrin N-glycans expressed in the milk of transgenic cows. 28170415 6 44 theme complex 758:764 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 10 45 dep % 1344:1344 arg1 fucosylation 1346:1357 fucosylation 1346:1357 20% fucosylation 1342:1357 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 2 46 theme N-glycosidic 211:222 arg1 bonds 224:228 N-glycosidic bonds 211:228 N-glycosidic bonds 211:228 Glycans attached through N-glycosidic bonds may contribute to Lactoferrin functional activities. 28170415 5 47 theme recombinant 533:543 arg1 rhLF 564:567 rhLF 564:567 rhLF 564:567 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 47 theme recombinant 533:543 arg1 lactoferrin 551:561 recombinant human lactoferrin 533:561 recombinant human lactoferrin (rhLF) 533:568 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 10 48 gly fucosylated 1286:1296 arg1 majority 1255:1262 the majority 1251:1262 the majority of hLF N-glycans 1251:1279 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 7 49 contain had 838:840 arg1 hLF 834:836 hLF 834:836 hLF 834:836 rhLF, bLF and hLF had 23, 27 and 18 N-glycans respectively with 8 N-glycan in common overall. 28170415 7 49 contain had 838:840 arg2 N-glycans 856:864 23, 27 and 18 N-glycans 842:864 23, 27 and 18 N-glycans 842:864 rhLF, bLF and hLF had 23, 27 and 18 N-glycans respectively with 8 N-glycan in common overall. 28170415 7 49 contain had 838:840 arg1 bLF 826:828 bLF 826:828 bLF 826:828 rhLF, bLF and hLF had 23, 27 and 18 N-glycans respectively with 8 N-glycan in common overall. 28170415 7 49 contain had 838:840 arg1 rhLF 820:823 rhLF 820:823 rhLF 820:823 rhLF, bLF and hLF had 23, 27 and 18 N-glycans respectively with 8 N-glycan in common overall. 28170415 4 50 theme alternative 402:412 arg1 approaches 414:423 alternative approaches 402:423 alternative approaches using transgenic cows to express human lactoferrin (hLF) 402:480 Efforts to increase LF concentration in bovine milk led to alternative approaches using transgenic cows to express human lactoferrin (hLF). 28170415 5 51 theme human 545:549 arg1 rhLF 564:567 rhLF 564:567 rhLF 564:567 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 51 theme human 545:549 arg1 lactoferrin 551:561 recombinant human lactoferrin 533:561 recombinant human lactoferrin (rhLF) 533:568 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 9 52 contain contain 1087:1093 arg2 N-glycans 1125:1133 mostly neutral complex/hybrid N-glycans 1095:1133 mostly neutral complex/hybrid N-glycans 1095:1133 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 52 contain contain 1087:1093 arg1 hLF 1071:1073 hLF 1071:1073 hLF 1071:1073 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 9 52 contain contain 1087:1093 arg1 rhLF 1062:1065 rhLF 1062:1065 rhLF 1062:1065 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 2 53 theme functional 260:269 arg1 activities 271:280 Lactoferrin functional activities 248:280 Lactoferrin functional activities 248:280 Glycans attached through N-glycosidic bonds may contribute to Lactoferrin functional activities. 28170415 6 54 theme fucosylated 731:741 arg1 glycans 766:772 fucosylated and sialylated complex glycans 731:772 glycans 766:772 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 9 55 theme neutral 1102:1108 arg1 N-glycans 1125:1133 mostly neutral complex/hybrid N-glycans 1095:1133 mostly neutral complex/hybrid N-glycans 1095:1133 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 3 56 from amounts 319:325 arg1 milk 337:340 bovine milk 330:340 bovine milk 330:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 56 from amounts 319:325 arg1 present 302:308 present 302:308 present 302:308 In contrast, LF is present in trace amounts in bovine milk. 28170415 2 57 theme Lactoferrin 248:258 arg1 activities 271:280 Lactoferrin functional activities 248:280 Lactoferrin functional activities 248:280 Glycans attached through N-glycosidic bonds may contribute to Lactoferrin functional activities. 28170415 10 58 theme N-glycans 1271:1279 arg1 majority 1255:1262 the majority 1251:1262 the majority of hLF N-glycans 1251:1279 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 9 59 theme types 1055:1059 arg1 abundances 1032:1041 the relative abundances 1019:1041 the relative abundances of N-glycan types 1019:1059 Based on the relative abundances of N-glycan types, rhLF and hLF appeared to contain mostly neutral complex/hybrid N-glycans (81% and 52% of the total respectively) whereas bLF was characterized by high mannose glycans (65%). 28170415 8 60 with N-glycan 953:960 arg1 hLF 967:969 hLF 967:969 hLF 967:969 rhLF shared 16 N-glycan with bLF and 9 N-glycan with hLF while bLF shared 10 N-glycan with hLF. 28170415 10 61 theme hLF 1267:1269 arg1 N-glycans 1271:1279 hLF N-glycans 1267:1279 hLF N-glycans 1267:1279 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 5 62 from N-glycans 520:528 arg1 lactoferrin 578:588 bovine lactoferrin 571:588 bovine lactoferrin (bLF) 571:594 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 62 from N-glycans 520:528 arg1 lactoferrin 606:616 human lactoferrin 600:616 human lactoferrin 600:616 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 62 from N-glycans 520:528 arg1 bLF 591:593 bLF 591:593 bLF 591:593 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 62 from N-glycans 520:528 arg1 rhLF 564:567 rhLF 564:567 rhLF 564:567 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 5 62 from N-glycans 520:528 arg1 lactoferrin 551:561 recombinant human lactoferrin 533:561 recombinant human lactoferrin (rhLF) 533:568 This study investigated and compared N-glycans in recombinant human lactoferrin (rhLF), bovine lactoferrin (bLF) and human lactoferrin by Nano-LC-Chip-Q-TOF Mass Spectrometry. 28170415 6 63 theme structures 709:718 arg1 diversity 687:695 a high diversity 680:695 a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities 680:817 The results revealed a high diversity of N-glycan structures, including fucosylated and sialylated complex glycans that may contribute additional bioactivities. 28170415 10 64 dep fucosylated 1286:1296 arg1 % 1301:1301 88% 1299:1301 88% 1299:1301 Interestingly, the majority of hLF N-glycans were fucosylated (88%), whereas bLF and rhLF had only 9% and 20% fucosylation, respectively. 28170415 3 65 from present 302:308 arg1 amounts 319:325 trace amounts 313:325 trace amounts in bovine milk 313:340 In contrast, LF is present in trace amounts in bovine milk. 28170415 3 65 from present 302:308 arg1 contrast 286:293 contrast 286:293 contrast 286:293 In contrast, LF is present in trace amounts in bovine milk. 26185202 7 0 theme IgG 990:992 arg1 glycans 994:1000 the IgG glycans 986:1000 the IgG glycans 986:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 6 1 from function 754:761 arg1 P<6.5×10 788:795 P<6.5×10(-4) 788:799 P<6.5×10(-4) 788:799 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 6 1 from function 754:761 arg1 sample 780:785 the discovery sample 766:785 the discovery sample (P<6.5×10(-4)) 766:800 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 7 2 theme glycosylation 886:898 arg1 sialylation 927:937 sialylation 927:937 sialylation 927:937 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 2 theme glycosylation 886:898 arg1 level 944:948 level 944:948 level 944:948 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 2 theme glycosylation 886:898 arg1 galactosylation 910:924 galactosylation 910:924 galactosylation 910:924 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 2 theme glycosylation 886:898 arg1 features 900:907 three main glycosylation features 875:907 three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans 875:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 1 3 theme damage 250:255 arg1 mechanisms 230:239 mechanisms 230:239 mechanisms of renal damage 230:255 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 8 4 theme diagnostic 1143:1152 arg1 targets 1170:1176 potential diagnostic and therapeutic targets 1133:1176 potential diagnostic and therapeutic targets 1133:1176 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 0 5 from Profile 14:20 arg1 Dysfunction 48:58 Moderate Kidney Dysfunction 32:58 Moderate Kidney Dysfunction 32:58 Glycosylation Profile of IgG in Moderate Kidney Dysfunction. 26185202 7 6 theme N-acetylglucosamine 963:981 arg1 sialylation 927:937 sialylation 927:937 sialylation 927:937 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 6 theme N-acetylglucosamine 963:981 arg1 level 944:948 level 944:948 level 944:948 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 6 theme N-acetylglucosamine 963:981 arg1 galactosylation 910:924 galactosylation 910:924 galactosylation 910:924 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 6 theme N-acetylglucosamine 963:981 arg1 features 900:907 three main glycosylation features 875:907 three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans 875:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 1 7 theme animal 160:165 arg1 studies 167:173 animal studies 160:173 animal studies 160:173 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 8 8 theme glycosylation 1038:1050 arg1 role 1026:1029 the role 1022:1029 the role of IgG glycosylation in kidney function 1022:1069 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 8 9 theme therapeutic 1158:1168 arg1 targets 1170:1176 potential diagnostic and therapeutic targets 1133:1176 potential diagnostic and therapeutic targets 1133:1176 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 6 10 theme discovery 770:778 arg1 P<6.5×10 788:795 P<6.5×10(-4) 788:799 P<6.5×10(-4) 788:799 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 6 10 theme discovery 770:778 arg1 sample 780:785 the discovery sample 766:785 the discovery sample (P<6.5×10(-4)) 766:800 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 3 11 theme larger 547:552 arg1 population 554:563 the larger population 543:563 the larger population 543:563 We analyzed the correlation between renal function measured as eGFR and 76 N-glycan traits using linear regressions adjusted for covariates and multiple testing in the larger population. 26185202 1 12 theme post-translational 122:139 arg1 modifications 141:153 the post-translational modifications 118:153 the post-translational modifications 118:153 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 8 13 theme kidney 1055:1060 arg1 function 1062:1069 kidney function 1055:1069 kidney function 1055:1069 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 7 14 theme bisecting 953:961 arg1 N-acetylglucosamine 963:981 bisecting N-acetylglucosamine 953:981 bisecting N-acetylglucosamine of the IgG glycans 953:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 6 15 theme glycan 713:718 arg1 traits 720:725 Fourteen glycan traits 704:725 Fourteen glycan traits 704:725 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 8 16 theme IgG 1034:1036 arg1 glycosylation 1038:1050 IgG glycosylation 1034:1050 IgG glycosylation 1034:1050 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 7 17 theme glycan 851:856 arg1 traits 858:863 Those glycan traits 845:863 Those glycan traits 845:863 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 0 18 theme Glycosylation 0:12 arg1 Profile 14:20 Glycosylation Profile 0:20 Glycosylation Profile of IgG in Moderate Kidney Dysfunction 0:58 Glycosylation Profile of IgG in Moderate Kidney Dysfunction. 26185202 1 19 theme abundant 89:96 arg1 form 110:113 the most abundant and diverse form 80:113 the most abundant and diverse form of the post-translational modifications 80:153 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 0 20 theme IgG 25:27 arg1 Profile 14:20 Glycosylation Profile 0:20 Glycosylation Profile of IgG in Moderate Kidney Dysfunction 0:58 Glycosylation Profile of IgG in Moderate Kidney Dysfunction. 26185202 8 21 theme novel 1083:1087 arg1 insight 1089:1095 novel insight 1083:1095 novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets 1083:1176 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 2 22 theme IgG 301:303 arg1 glycans 305:311 IgG glycans 301:311 IgG glycans 301:311 Here, we explored the associations between IgG glycans and renal function in 3274 individuals from the TwinsUK registry. 26185202 7 23 dep features 900:907 arg1 sialylation 927:937 sialylation 927:937 sialylation 927:937 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 23 dep features 900:907 arg1 level 944:948 level 944:948 level 944:948 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 23 dep features 900:907 arg1 galactosylation 910:924 galactosylation 910:924 galactosylation 910:924 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 23 dep features 900:907 arg1 features 900:907 three main glycosylation features 875:907 three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans 875:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 3 24 theme N-glycan 454:461 arg1 traits 463:468 76 N-glycan traits 451:468 76 N-glycan traits 451:468 We analyzed the correlation between renal function measured as eGFR and 76 N-glycan traits using linear regressions adjusted for covariates and multiple testing in the larger population. 26185202 6 25 theme renal 748:752 arg1 function 754:761 renal function 748:761 renal function in the discovery sample (P<6.5×10(-4)) 748:800 Fourteen glycan traits were associated with renal function in the discovery sample (P<6.5×10(-4)) and remained significant after validation. 26185202 1 26 theme diverse 102:108 arg1 form 110:113 the most abundant and diverse form 80:113 the most abundant and diverse form of the post-translational modifications 80:153 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 1 27 theme modifications 141:153 arg1 form 110:113 the most abundant and diverse form 80:113 the most abundant and diverse form of the post-translational modifications 80:153 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 0 28 theme Moderate 32:39 arg1 Dysfunction 48:58 Moderate Kidney Dysfunction 32:58 Moderate Kidney Dysfunction 32:58 Glycosylation Profile of IgG in Moderate Kidney Dysfunction. 26185202 8 29 from role 1026:1029 arg1 function 1062:1069 kidney function 1055:1069 kidney function 1055:1069 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 8 30 gly glycosylation 1038:1050 arg1 kidney 1055:1060 kidney function 1055:1069 kidney function 1055:1069 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 1 31 theme IgG 209:211 arg1 glycosylation 213:225 IgG glycosylation 209:225 IgG glycosylation 209:225 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 3 32 theme renal 415:419 arg1 function 421:428 renal function 415:428 renal function measured as eGFR and 76 N-glycan traits 415:468 We analyzed the correlation between renal function measured as eGFR and 76 N-glycan traits using linear regressions adjusted for covariates and multiple testing in the larger population. 26185202 1 33 theme glycosylation 213:225 arg1 involvement 194:204 the involvement 190:204 the involvement of IgG glycosylation in mechanisms of renal damage 190:255 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 5 34 from analyses 670:677 arg1 Results 652:658 Results 652:658 Results from both analyses 652:677 Results from both analyses were then meta-analyzed. 26185202 2 35 from registry 369:376 arg1 individuals 340:350 3274 individuals 335:350 3274 individuals from the TwinsUK registry 335:376 Here, we explored the associations between IgG glycans and renal function in 3274 individuals from the TwinsUK registry. 26185202 0 36 theme Kidney 41:46 arg1 Dysfunction 48:58 Moderate Kidney Dysfunction 32:58 Moderate Kidney Dysfunction 32:58 Glycosylation Profile of IgG in Moderate Kidney Dysfunction. 26185202 1 37 from involvement 194:204 arg1 mechanisms 230:239 mechanisms 230:239 mechanisms of renal damage 230:255 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 26185202 8 38 theme CKD 1125:1127 arg1 pathophysiology 1106:1120 the pathophysiology 1102:1120 the pathophysiology of CKD 1102:1127 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 8 38 theme CKD 1125:1127 arg1 targets 1170:1176 potential diagnostic and therapeutic targets 1133:1176 potential diagnostic and therapeutic targets 1133:1176 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 4 39 theme twin 610:613 arg1 pairs 615:619 31 monozygotic twin pairs 595:619 31 monozygotic twin pairs discordant for renal function 595:649 We replicated our results in 31 monozygotic twin pairs discordant for renal function. 26185202 7 40 theme glycans 994:1000 arg1 N-acetylglucosamine 963:981 bisecting N-acetylglucosamine 953:981 bisecting N-acetylglucosamine of the IgG glycans 953:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 2 41 theme renal 317:321 arg1 function 323:330 renal function 317:330 renal function 317:330 Here, we explored the associations between IgG glycans and renal function in 3274 individuals from the TwinsUK registry. 26185202 4 42 theme monozygotic 598:608 arg1 pairs 615:619 31 monozygotic twin pairs 595:619 31 monozygotic twin pairs discordant for renal function 595:649 We replicated our results in 31 monozygotic twin pairs discordant for renal function. 26185202 8 43 theme potential 1133:1141 arg1 targets 1170:1176 potential diagnostic and therapeutic targets 1133:1176 potential diagnostic and therapeutic targets 1133:1176 These results show the role of IgG glycosylation in kidney function and provide novel insight into the pathophysiology of CKD and potential diagnostic and therapeutic targets. 26185202 4 44 theme renal 636:640 arg1 function 642:649 renal function 636:649 renal function 636:649 We replicated our results in 31 monozygotic twin pairs discordant for renal function. 26185202 2 45 theme TwinsUK 361:367 arg1 registry 369:376 the TwinsUK registry 357:376 the TwinsUK registry 357:376 Here, we explored the associations between IgG glycans and renal function in 3274 individuals from the TwinsUK registry. 26185202 3 46 theme linear 476:481 arg1 regressions 483:493 linear regressions 476:493 linear regressions adjusted for covariates and multiple testing in the larger population 476:563 We analyzed the correlation between renal function measured as eGFR and 76 N-glycan traits using linear regressions adjusted for covariates and multiple testing in the larger population. 26185202 4 47 theme discordant 621:630 arg1 pairs 615:619 31 monozygotic twin pairs 595:619 31 monozygotic twin pairs discordant for renal function 595:649 We replicated our results in 31 monozygotic twin pairs discordant for renal function. 26185202 3 48 theme multiple 523:530 arg1 testing 532:538 multiple testing 523:538 multiple testing 523:538 We analyzed the correlation between renal function measured as eGFR and 76 N-glycan traits using linear regressions adjusted for covariates and multiple testing in the larger population. 26185202 7 49 theme main 881:884 arg1 sialylation 927:937 sialylation 927:937 sialylation 927:937 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 49 theme main 881:884 arg1 level 944:948 level 944:948 level 944:948 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 49 theme main 881:884 arg1 galactosylation 910:924 galactosylation 910:924 galactosylation 910:924 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 7 49 theme main 881:884 arg1 features 900:907 three main glycosylation features 875:907 three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans 875:1000 Those glycan traits belong to three main glycosylation features: galactosylation, sialylation, and level of bisecting N-acetylglucosamine of the IgG glycans. 26185202 1 50 theme renal 244:248 arg1 damage 250:255 renal damage 244:255 renal damage 244:255 Glycans constitute the most abundant and diverse form of the post-translational modifications, and animal studies have suggested the involvement of IgG glycosylation in mechanisms of renal damage. 28079948 1 0 attach attachment 174:183 arg2 α-mannopyranose 188:202 α-mannopyranose 188:202 α-mannopyranose 188:202 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 0 attach attachment 174:183 arg1 Thr 214:216 Thr 214:216 Thr 214:216 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 0 attach attachment 174:183 arg1 Ser 207:209 Ser 207:209 Ser 207:209 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 1 theme protein 136:142 arg1 O-mannosylation 144:158 Mammalian protein O-mannosylation 126:158 Mammalian protein O-mannosylation 126:158 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 3 2 theme glycoproteome 578:590 arg1 impact 543:548 the biological impact 528:548 the biological impact of the mammalian O-mannosyl glycoproteome 528:590 Despite the enormous progress made, the biological impact of the mammalian O-mannosyl glycoproteome remains largely unknown to date. 28079948 9 3 theme molecular 1748:1756 arg1 structure 1758:1766 molecular structure 1748:1766 molecular structure 1748:1766 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 0 4 theme Complimentary 92:104 arg1 Glycopeptides-Selectivity 66:90 Branched Core O-Mannosyl Glycopeptides-Selectivity 41:90 Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin 41:123 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 4 5 theme glycans 712:718 arg1 role 678:681 role 678:681 role 678:681 Tools are still needed to investigate the structure, role, and abundance of O-mannosyl glycans. 28079948 4 5 theme glycans 712:718 arg1 abundance 688:696 abundance 688:696 abundance 688:696 Tools are still needed to investigate the structure, role, and abundance of O-mannosyl glycans. 28079948 4 5 theme glycans 712:718 arg1 structure 667:675 structure 667:675 structure 667:675 Tools are still needed to investigate the structure, role, and abundance of O-mannosyl glycans. 28079948 2 6 theme Recent 330:335 arg1 advances 337:344 Recent advances 330:344 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy 330:405 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 5 7 theme broader 902:908 arg1 understanding 910:922 a broader understanding 900:922 a broader understanding of the biological roles of branched O-mannosyl glycans 900:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 5 8 from relevance 775:783 arg1 migration 812:820 integrin-dependent cell migration 788:820 integrin-dependent cell migration 788:820 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 10 9 theme ConA 1854:1857 arg1 specificity 1867:1877 the ConA binding specificity 1850:1877 the ConA binding specificity 1850:1877 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 6 10 used used 1104:1107 arg2 construct 1074:1082 a glycopeptide vaccine construct 1051:1082 a glycopeptide vaccine construct 1051:1082 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 5 11 theme multiple 880:887 arg1 sclerosis 889:897 multiple sclerosis 880:897 multiple sclerosis 880:897 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 4 12 dep structure 667:675 arg1 the 663:665 the 663:665 the 663:665 Tools are still needed to investigate the structure, role, and abundance of O-mannosyl glycans. 28079948 5 13 theme O-mannosyl 960:969 arg1 glycans 971:977 branched O-mannosyl glycans 951:977 branched O-mannosyl glycans 951:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 5 14 theme cell 807:810 arg1 migration 812:820 integrin-dependent cell migration 788:820 integrin-dependent cell migration 788:820 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 0 15 theme Antibodies 13:22 arg1 Induction 0:8 Induction 0:8 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 0:124 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 8 16 theme O-mannosyl 1556:1565 arg1 peptides 1567:1574 tryptic O-mannosyl peptides 1548:1574 tryptic O-mannosyl peptides 1548:1574 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 2 17 gly glycoproteins 444:456 arg1 glycoproteins 444:456 glycoproteins 444:456 glycoproteins 444:456 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 7 18 theme glycan 1268:1273 arg1 structures 1280:1289 branched glycan core structures 1259:1289 branched glycan core structures presented on different peptide backbones 1259:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 1 19 theme α-mannopyranose 188:202 arg1 attachment 174:183 attachment 174:183 attachment of α-mannopyranose to Ser or Thr residues 174:225 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 2 20 from advances 337:344 arg1 methodology 365:375 glycoproteomics methodology 349:375 glycoproteomics methodology 349:375 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 2 20 from advances 337:344 arg1 " 396:396 the "SimpleCell" 381:396 the "SimpleCell" 381:396 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 1 21 dep muscle 300:305 arg1 development 317:327 development 317:327 development 317:327 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 8 22 theme mannose-binding 1440:1454 arg1 ConA 1479:1482 ConA 1479:1482 ConA 1479:1482 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 8 22 theme mannose-binding 1440:1454 arg1 A 1476:1476 the mannose-binding lectin concanavalin A 1436:1476 the mannose-binding lectin concanavalin A (ConA) 1436:1483 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 0 23 theme ConA 113:116 arg1 Lectin 118:123 the ConA Lectin 109:123 the ConA Lectin 109:123 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 5 24 theme due 998:1000 arg1 part 993:996 part 993:996 part due to the paucity of detection tools 993:1034 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 6 25 theme glycopeptide 1053:1064 arg1 construct 1074:1082 a glycopeptide vaccine construct 1051:1082 a glycopeptide vaccine construct 1051:1082 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 2 26 theme rapid 420:424 arg1 identification 426:439 rapid identification 420:439 rapid identification of glycoproteins and specific glycosylation sites 420:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 7 27 theme induced 1236:1242 arg1 antibodies 1244:1253 the induced antibodies 1232:1253 the induced antibodies 1232:1253 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 0 28 theme Branched 41:48 arg1 Glycopeptides-Selectivity 66:90 Branched Core O-Mannosyl Glycopeptides-Selectivity 41:90 Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin 41:123 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 2 29 gly glycosylation 471:483 arg2 sites 485:489 specific glycosylation sites 462:489 specific glycosylation sites 462:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 8 30 theme ConA 1593:1596 arg1 lectin 1598:1603 the ConA lectin 1589:1603 the ConA lectin 1589:1603 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 7 31 theme core 1275:1278 arg1 structures 1280:1289 branched glycan core structures 1259:1289 branched glycan core structures presented on different peptide backbones 1259:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 2 32 theme glycosylation 471:483 arg1 sites 485:489 specific glycosylation sites 462:489 specific glycosylation sites 462:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 6 33 gly glycopeptide 1053:1064 arg2 glycopeptide 1053:1064 a glycopeptide vaccine construct 1051:1082 a glycopeptide vaccine construct 1051:1082 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 8 34 theme lectin 1456:1461 arg1 ConA 1479:1482 ConA 1479:1482 ConA 1479:1482 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 8 34 theme lectin 1456:1461 arg1 A 1476:1476 the mannose-binding lectin concanavalin A 1436:1476 the mannose-binding lectin concanavalin A (ConA) 1436:1483 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 3 35 theme enormous 504:511 arg1 progress 513:520 the enormous progress 500:520 the enormous progress made 500:525 Despite the enormous progress made, the biological impact of the mammalian O-mannosyl glycoproteome remains largely unknown to date. 28079948 7 36 theme high 1212:1215 arg1 selectivity 1217:1227 high selectivity 1212:1227 high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones 1212:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 1 37 theme Mammalian 126:134 arg1 O-mannosylation 144:158 Mammalian protein O-mannosylation 126:158 Mammalian protein O-mannosylation 126:158 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 7 38 theme related 1372:1378 arg1 glycans 1387:1393 related linear glycans 1372:1393 related linear glycans 1372:1393 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 8 39 theme A 1476:1476 arg1 screening 1423:1431 microarray screening 1412:1431 microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, 1412:1575 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 8 40 theme O-mannosyl 1632:1641 arg1 glycans 1643:1649 branched O-mannosyl glycans 1623:1649 branched O-mannosyl glycans 1623:1649 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 7 41 theme Glycopeptide 1169:1180 arg1 screening 1193:1201 Glycopeptide microarray screening 1169:1201 Glycopeptide microarray screening 1169:1201 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 9 42 theme ConA 1678:1681 arg1 preference 1664:1673 The binding preference 1652:1673 The binding preference of ConA for short linear O-mannosyl glycans 1652:1717 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 6 43 theme O-mannosyl 1149:1158 arg1 glycans 1160:1166 branched O-mannosyl glycans 1140:1166 branched O-mannosyl glycans 1140:1166 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 2 44 theme glycoproteomics 349:363 arg1 methodology 365:375 glycoproteomics methodology 349:375 glycoproteomics methodology 349:375 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 9 45 theme short 1687:1691 arg1 glycans 1711:1717 short linear O-mannosyl glycans 1687:1717 short linear O-mannosyl glycans 1687:1717 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 1 46 theme post-translational 248:265 arg1 PTMs 282:285 PTMs 282:285 PTMs 282:285 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 46 theme post-translational 248:265 arg1 modifications 267:279 post-translational modifications 248:279 post-translational modifications (PTMs) involved in muscle and brain development 248:327 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 10 47 from role 1928:1931 arg1 studies 1955:1961 studies 1955:1961 studies of protein O-mannosylation 1955:1988 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 9 48 theme O-mannosyl 1700:1709 arg1 glycans 1711:1717 short linear O-mannosyl glycans 1687:1717 short linear O-mannosyl glycans 1687:1717 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 7 49 theme peptide 1314:1320 arg1 backbones 1322:1330 different peptide backbones 1304:1330 different peptide backbones 1304:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 10 50 theme O-mannosylation 1974:1988 arg1 studies 1955:1961 studies 1955:1961 studies of protein O-mannosylation 1955:1988 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 3 51 theme O-mannosyl 567:576 arg1 glycoproteome 578:590 the mammalian O-mannosyl glycoproteome 553:590 the mammalian O-mannosyl glycoproteome 553:590 Despite the enormous progress made, the biological impact of the mammalian O-mannosyl glycoproteome remains largely unknown to date. 28079948 0 52 theme Core 50:53 arg1 Glycopeptides-Selectivity 66:90 Branched Core O-Mannosyl Glycopeptides-Selectivity 41:90 Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin 41:123 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 9 53 theme structure 1758:1766 arg1 terms 1739:1743 terms 1739:1743 terms of molecular structure 1739:1766 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 4 54 theme O-mannosyl 701:710 arg1 glycans 712:718 O-mannosyl glycans 701:718 O-mannosyl glycans 701:718 Tools are still needed to investigate the structure, role, and abundance of O-mannosyl glycans. 28079948 10 55 theme binding 1859:1865 arg1 specificity 1867:1877 the ConA binding specificity 1850:1877 the ConA binding specificity 1850:1877 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 9 56 theme crystallographic 1774:1789 arg1 data 1791:1794 crystallographic data 1774:1794 crystallographic data augmented by molecular modeling 1774:1826 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 5 57 theme biological 931:940 arg1 roles 942:946 the biological roles 927:946 the biological roles of branched O-mannosyl glycans 927:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 8 58 used used 1504:1507 arg2 ConA 1479:1482 ConA 1479:1482 ConA 1479:1482 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 8 58 used used 1504:1507 arg2 A 1476:1476 the mannose-binding lectin concanavalin A 1436:1476 the mannose-binding lectin concanavalin A (ConA) 1436:1483 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 10 59 theme new 1895:1897 arg1 antibodies 1899:1908 the new antibodies 1891:1908 the new antibodies 1891:1908 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 8 60 theme glycoproteomics 1512:1526 arg1 workflows 1528:1536 glycoproteomics workflows 1512:1536 glycoproteomics workflows 1512:1536 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 5 61 theme roles 942:946 arg1 understanding 910:922 a broader understanding 900:922 a broader understanding of the biological roles of branched O-mannosyl glycans 900:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 9 62 theme molecular 1809:1817 arg1 modeling 1819:1826 molecular modeling 1809:1826 molecular modeling 1809:1826 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 5 63 theme O-mannosyl 730:739 arg1 branching 741:749 O-mannosyl branching 730:749 O-mannosyl branching 730:749 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 10 64 theme novel 1922:1926 arg1 role 1928:1931 a novel role 1920:1931 a novel role for the antibodies in studies of protein O-mannosylation 1920:1988 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 5 65 theme branched 951:958 arg1 glycans 971:977 branched O-mannosyl glycans 951:977 branched O-mannosyl glycans 951:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 8 66 theme tryptic 1548:1554 arg1 peptides 1567:1574 tryptic O-mannosyl peptides 1548:1574 tryptic O-mannosyl peptides 1548:1574 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 5 67 theme integrin-dependent 788:805 arg1 migration 812:820 integrin-dependent cell migration 788:820 integrin-dependent cell migration 788:820 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 7 68 theme branched 1259:1266 arg1 structures 1280:1289 branched glycan core structures 1259:1289 branched glycan core structures presented on different peptide backbones 1259:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 7 69 theme antibodies 1244:1253 arg1 selectivity 1217:1227 high selectivity 1212:1227 high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones 1212:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 8 70 theme microarray 1412:1421 arg1 screening 1423:1431 microarray screening 1412:1431 microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, 1412:1575 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 5 71 theme glycans 971:977 arg1 roles 942:946 the biological roles 927:946 the biological roles of branched O-mannosyl glycans 927:977 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 7 72 theme linear 1380:1385 arg1 glycans 1387:1393 related linear glycans 1372:1393 related linear glycans 1372:1393 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 2 73 theme sites 485:489 arg1 identification 426:439 rapid identification 420:439 rapid identification of glycoproteins and specific glycosylation sites 420:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 5 74 theme demyelinating 848:860 arg1 sclerosis 889:897 multiple sclerosis 880:897 multiple sclerosis 880:897 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 5 74 theme demyelinating 848:860 arg1 diseases 862:869 demyelinating diseases 848:869 demyelinating diseases 848:869 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 9 75 theme linear 1693:1698 arg1 glycans 1711:1717 short linear O-mannosyl glycans 1687:1717 short linear O-mannosyl glycans 1687:1717 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 6 76 theme vaccine 1066:1072 arg1 construct 1074:1082 a glycopeptide vaccine construct 1051:1082 a glycopeptide vaccine construct 1051:1082 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 2 77 theme specific 462:469 arg1 sites 485:489 specific glycosylation sites 462:489 specific glycosylation sites 462:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 8 78 theme branched 1623:1630 arg1 glycans 1643:1649 branched O-mannosyl glycans 1623:1649 branched O-mannosyl glycans 1623:1649 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 2 79 dep methodology 365:375 arg1 strategy 398:405 strategy 398:405 strategy 398:405 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 2 80 theme glycoproteins 444:456 arg1 identification 426:439 rapid identification 420:439 rapid identification of glycoproteins and specific glycosylation sites 420:489 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 7 81 theme microarray 1182:1191 arg1 screening 1193:1201 Glycopeptide microarray screening 1169:1201 Glycopeptide microarray screening 1169:1201 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 9 82 theme binding 1656:1662 arg1 preference 1664:1673 The binding preference 1652:1673 The binding preference of ConA for short linear O-mannosyl glycans 1652:1717 The binding preference of ConA for short linear O-mannosyl glycans was rationalized in terms of molecular structure using crystallographic data augmented by molecular modeling. 28079948 8 83 theme concanavalin 1463:1474 arg1 ConA 1479:1482 ConA 1479:1482 ConA 1479:1482 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 8 83 theme concanavalin 1463:1474 arg1 A 1476:1476 the mannose-binding lectin concanavalin A 1436:1476 the mannose-binding lectin concanavalin A (ConA) 1436:1483 For comparison, microarray screening of the mannose-binding lectin concanavalin A (ConA), which is commonly used in glycoproteomics workflows to enrich tryptic O-mannosyl peptides, showed that the ConA lectin did not recognize branched O-mannosyl glycans. 28079948 0 84 theme O-Mannosyl 55:64 arg1 Glycopeptides-Selectivity 66:90 Branched Core O-Mannosyl Glycopeptides-Selectivity 41:90 Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin 41:123 Induction of Antibodies Directed Against Branched Core O-Mannosyl Glycopeptides-Selectivity Complimentary to the ConA Lectin. 28079948 10 85 theme protein 1966:1972 arg1 O-mannosylation 1974:1988 protein O-mannosylation 1966:1988 protein O-mannosylation 1966:1988 The contrast between the ConA binding specificity and that of the new antibodies indicates a novel role for the antibodies in studies of protein O-mannosylation. 28079948 1 86 dep Ser 207:209 arg1 residues 218:225 residues 218:225 residues 218:225 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 3 87 theme biological 532:541 arg1 impact 543:548 the biological impact 528:548 the biological impact of the mammalian O-mannosyl glycoproteome 528:590 Despite the enormous progress made, the biological impact of the mammalian O-mannosyl glycoproteome remains largely unknown to date. 28079948 5 88 theme tools 1030:1034 arg1 paucity 1009:1015 the paucity 1005:1015 the paucity of detection tools 1005:1034 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 6 89 theme branched 1140:1147 arg1 glycans 1160:1166 branched O-mannosyl glycans 1140:1166 branched O-mannosyl glycans 1140:1166 In this work, a glycopeptide vaccine construct was synthesized and used to generate antibodies against branched O-mannosyl glycans. 28079948 2 90 theme SimpleCell 386:395 arg1 " 396:396 the "SimpleCell" 381:396 the "SimpleCell" 381:396 Recent advances in glycoproteomics methodology and the "SimpleCell" strategy have enabled rapid identification of glycoproteins and specific glycosylation sites. 28079948 5 91 theme detection 1020:1028 arg1 tools 1030:1034 detection tools 1020:1034 detection tools 1020:1034 Although O-mannosyl branching has been shown to be of relevance in integrin-dependent cell migration, and also plays a role in demyelinating diseases, such as multiple sclerosis, a broader understanding of the biological roles of branched O-mannosyl glycans is lacking in part due to the paucity of detection tools. 28079948 7 92 theme different 1304:1312 arg1 backbones 1322:1330 different peptide backbones 1304:1330 different peptide backbones 1304:1330 Glycopeptide microarray screening revealed high selectivity of the induced antibodies for branched glycan core structures presented on different peptide backbones, with no cross-reactivity observed with related linear glycans. 28079948 3 93 theme mammalian 557:565 arg1 glycoproteome 578:590 the mammalian O-mannosyl glycoproteome 553:590 the mammalian O-mannosyl glycoproteome 553:590 Despite the enormous progress made, the biological impact of the mammalian O-mannosyl glycoproteome remains largely unknown to date. 28079948 1 94 theme modifications 267:279 arg1 PTMs 282:285 PTMs 282:285 PTMs 282:285 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 94 theme modifications 267:279 arg1 group 239:243 a group 237:243 a group of post-translational modifications (PTMs) involved in muscle and brain development 237:327 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 28079948 1 94 theme modifications 267:279 arg1 modifications 267:279 post-translational modifications 248:279 post-translational modifications (PTMs) involved in muscle and brain development 248:327 Mammalian protein O-mannosylation, initiated by attachment of α-mannopyranose to Ser or Thr residues, comprise a group of post-translational modifications (PTMs) involved in muscle and brain development. 29288327 9 0 theme daughter 1690:1697 arg1 walls 1705:1709 daughter frond walls 1690:1709 daughter frond walls 1690:1709 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 3 1 theme plants 713:718 arg1 walls 683:687 the primary walls 671:687 the primary walls of most other flowering plants 671:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 1 2 theme cell 263:266 arg1 apiogalacturonan 273:288 cell wall apiogalacturonan 263:288 cell wall apiogalacturonan 263:288 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 3 3 theme large 610:614 arg1 apiogalacturonan 627:642 apiogalacturonan 627:642 apiogalacturonan 627:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 3 theme large 610:614 arg1 amounts 616:622 large amounts 610:622 large amounts of apiogalacturonan 610:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 7 4 theme terrestrial 1421:1431 arg1 plants 1433:1438 terrestrial plants 1421:1438 terrestrial plants 1421:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 3 5 theme primary 578:584 arg1 walls 591:595 primary cell walls 578:595 primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants 578:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 1 6 theme apiogalacturonan 273:288 arg1 abundance 250:258 the abundance 246:258 the abundance of cell wall apiogalacturonan 246:288 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 10 7 from changes 1959:1965 arg1 pectins 1976:1982 other pectins 1970:1982 other pectins 1970:1982 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 11 8 theme model 2010:2014 arg1 system 2016:2021 a model system 2008:2021 a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits 2008:2150 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 3 9 from walls 683:687 arg1 distinct 657:664 distinct 657:664 distinct 657:664 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 9 10 theme frond 1726:1730 arg1 walls 1732:1736 mother frond walls 1719:1736 mother frond walls 1719:1736 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 7 11 theme similar 1399:1405 arg1 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 11 theme similar 1399:1405 arg1 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 10 12 theme other 1970:1974 arg1 pectins 1976:1982 other pectins 1970:1982 other pectins 1970:1982 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 1 13 theme Lemnoideae 201:210 arg1 diversification 178:192 The diversification 174:192 The diversification of the Lemnoideae 174:210 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 7 14 theme glycosyl 1381:1388 arg1 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 14 theme glycosyl 1381:1388 arg1 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 11 15 theme related 2110:2116 arg1 plants 2118:2123 closely related plants 2102:2123 closely related plants with similar growth habits 2102:2150 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 10 16 theme cell 1859:1862 arg1 wall 1864:1867 a functional cell wall 1846:1867 a functional cell wall 1846:1867 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 11 17 theme growth 2138:2143 arg1 habits 2145:2150 similar growth habits 2130:2150 similar growth habits 2130:2150 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 7 18 theme borate 1345:1350 arg1 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 18 theme borate 1345:1350 arg1 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 6 19 theme arabinose 1185:1193 arg1 content 1195:1201 a high arabinose content 1178:1201 a high arabinose content 1178:1201 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 3 20 dep contain 602:608 arg1 distinct 657:664 distinct 657:664 distinct 657:664 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 0 21 from Changes 0:6 arg1 abundance 15:23 the abundance 11:23 the abundance of cell wall 11:36 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 8 22 theme sidechain 1532:1540 arg1 A 1542:1542 RG-II sidechain A 1526:1542 RG-II sidechain A 1526:1542 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 4 23 theme Lemnoideae 784:793 arg1 members 769:775 other members 763:775 other members of the Lemnoideae 763:793 However, the pectins in the cell walls of other members of the Lemnoideae have not been investigated. 29288327 7 24 contain has 1375:1377 arg1 RG-II 1306:1310 RG-II 1306:1310 RG-II 1306:1310 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 24 contain has 1375:1377 arg2 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 24 contain has 1375:1377 arg2 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 24 contain has 1375:1377 arg1 rhamnogalacturonan 1283:1300 The apiose-containing pectin rhamnogalacturonan II 1254:1303 The apiose-containing pectin rhamnogalacturonan II (RG-II) 1254:1311 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 5 25 dep Wolffiella 919:928 arg1 walls 942:946 walls 942:946 walls 942:946 Here, we show that apiogalacturonan decreased substantially as the Lemnoideae diversified since Wolffiella and Wolffia walls contain between 63 and 88% less apiose than Spirodela, Landoltia, and Lemna walls. 29288327 2 26 dep small 470:474 arg1 fast-growing 477:488 fast-growing 477:488 fast-growing 477:488 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 6 27 theme derived 1052:1058 arg1 genus 1060:1064 the most derived genus 1043:1064 the most derived genus 1043:1064 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 27 theme derived 1052:1058 arg1 abundant 1096:1103 abundant 1096:1103 abundant 1096:1103 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 27 theme derived 1052:1058 arg1 xylogalacturonan 1067:1082 xylogalacturonan 1067:1082 xylogalacturonan 1067:1082 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 4 28 theme other 763:767 arg1 members 769:775 other members 763:775 other members of the Lemnoideae 763:793 However, the pectins in the cell walls of other members of the Lemnoideae have not been investigated. 29288327 11 29 with plants 2118:2123 arg1 habits 2145:2150 similar growth habits 2130:2150 similar growth habits 2130:2150 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 8 30 theme B 1577:1577 arg1 A 1542:1542 RG-II sidechain A 1526:1542 RG-II sidechain A 1526:1542 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 8 30 theme B 1577:1577 arg1 arabinosylation 1548:1562 arabinosylation 1548:1562 arabinosylation of sidechain B 1548:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 10 31 theme cross-linking 1914:1926 arg1 conservation 1878:1889 conservation 1878:1889 conservation of RG-II structure and cross-linking 1878:1926 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 2 32 theme aquatic 490:496 arg1 monocots 498:505 small, fast-growing aquatic monocots 470:505 small, fast-growing aquatic monocots 470:505 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 10 33 theme structure 1900:1908 arg1 conservation 1878:1889 conservation 1878:1889 conservation of RG-II structure and cross-linking 1878:1926 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 0 34 theme wall 33:36 arg1 abundance 15:23 the abundance 11:23 the abundance of cell wall 11:36 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 1 35 theme rhamnogalacturonan 334:351 arg1 structure 356:364 rhamnogalacturonan II structure 334:364 rhamnogalacturonan II structure 334:364 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 3 36 contain have 573:576 arg1 Lemna 508:512 Lemna minor 508:518 Lemna minor 508:518 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 36 contain have 573:576 arg2 walls 591:595 primary cell walls 578:595 primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants 578:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 10 37 theme structural 1948:1957 arg1 changes 1959:1965 structural changes 1948:1965 structural changes in other pectins 1948:1982 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 7 38 theme apiose-containing 1258:1274 arg1 RG-II 1306:1310 RG-II 1306:1310 RG-II 1306:1310 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 38 theme apiose-containing 1258:1274 arg1 rhamnogalacturonan 1283:1300 The apiose-containing pectin rhamnogalacturonan II 1254:1303 The apiose-containing pectin rhamnogalacturonan II (RG-II) 1254:1311 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 3 39 theme most 692:695 arg1 plants 713:718 most other flowering plants 692:718 most other flowering plants 692:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 2 40 theme subfamily 403:411 arg1 Lemnoideae 413:422 The subfamily Lemnoideae 399:422 The subfamily Lemnoideae 399:422 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 6 41 link derived 1052:1058 arg1 genus 1060:1064 the most derived genus 1043:1064 the most derived genus 1043:1064 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 41 link derived 1052:1058 arg1 abundant 1096:1103 abundant 1096:1103 abundant 1096:1103 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 41 link derived 1052:1058 arg1 xylogalacturonan 1067:1082 xylogalacturonan 1067:1082 xylogalacturonan 1067:1082 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 9 42 theme frond 1699:1703 arg1 walls 1705:1709 daughter frond walls 1690:1709 daughter frond walls 1690:1709 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 1 43 theme wall 268:271 arg1 apiogalacturonan 273:288 cell wall apiogalacturonan 263:288 cell wall apiogalacturonan 263:288 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 6 44 theme high 1180:1183 arg1 content 1195:1201 a high arabinose content 1178:1201 a high arabinose content 1178:1201 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 1 45 from increase 297:304 arg1 abundance 250:258 the abundance 246:258 the abundance of cell wall apiogalacturonan 246:288 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 1 45 from increase 297:304 arg1 xylogalacturonan 309:324 xylogalacturonan 309:324 xylogalacturonan 309:324 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 11 46 theme wall 2056:2059 arg1 structure 2061:2069 wall structure 2056:2069 wall structure 2056:2069 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 9 47 theme mother 1719:1724 arg1 walls 1732:1736 mother frond walls 1719:1736 mother frond walls 1719:1736 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 6 48 dep in 1136:1137 arg1 Wolffiella 1139:1148 Wolffiella 1139:1148 Wolffiella 1139:1148 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 49 theme RG 1248:1249 arg1 sidechains 1234:1243 arabinan sidechains 1225:1243 arabinan sidechains of RG I 1225:1251 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 50 from Wolffia 1034:1040 arg1 genus 1060:1064 the most derived genus 1043:1064 the most derived genus 1043:1064 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 50 from Wolffia 1034:1040 arg1 abundant 1096:1103 abundant 1096:1103 abundant 1096:1103 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 51 theme pectic 1150:1155 arg1 polysaccharides 1157:1171 in Wolffiella pectic polysaccharides 1136:1171 in Wolffiella pectic polysaccharides 1136:1171 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 3 52 theme cell 586:589 arg1 walls 591:595 primary cell walls 578:595 primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants 578:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 11 53 theme similar 2130:2136 arg1 habits 2145:2150 similar growth habits 2130:2150 similar growth habits 2130:2150 The Lemnoideae provide a model system to study the mechanisms by which wall structure and composition has changed in closely related plants with similar growth habits. 29288327 6 54 theme in 1136:1137 arg1 polysaccharides 1157:1171 in Wolffiella pectic polysaccharides 1136:1171 in Wolffiella pectic polysaccharides 1136:1171 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 8 55 theme species-dependent 1455:1471 arg1 variations 1473:1482 species-dependent variations 1455:1482 species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B 1455:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 6 56 theme arabinan 1225:1232 arg1 sidechains 1234:1243 arabinan sidechains 1225:1243 arabinan sidechains of RG I 1225:1251 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 0 57 theme Lemnoideae 162:171 arg1 diversification 139:153 the diversification 135:153 the diversification of the Lemnoideae 135:171 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 10 58 theme functional 1848:1857 arg1 wall 1864:1867 a functional cell wall 1846:1867 a functional cell wall 1846:1867 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 3 59 theme flowering 703:711 arg1 plants 713:718 most other flowering plants 692:718 most other flowering plants 692:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 5 60 dep Spirodela 992:1000 arg1 walls 1024:1028 walls 1024:1028 walls 1024:1028 Here, we show that apiogalacturonan decreased substantially as the Lemnoideae diversified since Wolffiella and Wolffia walls contain between 63 and 88% less apiose than Spirodela, Landoltia, and Lemna walls. 29288327 1 61 from reduction 233:241 arg1 abundance 250:258 the abundance 246:258 the abundance of cell wall apiogalacturonan 246:288 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 1 61 from reduction 233:241 arg1 xylogalacturonan 309:324 xylogalacturonan 309:324 xylogalacturonan 309:324 The diversification of the Lemnoideae was accompanied by a reduction in the abundance of cell wall apiogalacturonan and an increase in xylogalacturonan whereas rhamnogalacturonan II structure and cross-linking are conserved. 29288327 10 62 theme RG-II 1894:1898 arg1 structure 1900:1908 RG-II structure 1894:1908 RG-II structure 1894:1908 Our data support the notion that a functional cell wall requires conservation of RG-II structure and cross-linking but can accommodate structural changes in other pectins. 29288327 9 63 theme expanding 1796:1804 arg1 cells 1806:1810 expanding cells 1796:1810 expanding cells 1796:1810 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 7 64 theme cross-linked 1352:1363 arg1 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 64 theme cross-linked 1352:1363 arg1 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 5 65 theme diversified 901:911 arg1 Lemnoideae 890:899 the Lemnoideae 886:899 the Lemnoideae diversified since Wolffiella and Wolffia walls 886:946 Here, we show that apiogalacturonan decreased substantially as the Lemnoideae diversified since Wolffiella and Wolffia walls contain between 63 and 88% less apiose than Spirodela, Landoltia, and Lemna walls. 29288327 8 66 theme methyl-etherification 1501:1521 arg1 extent 1491:1496 the extent 1487:1496 the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B 1487:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 8 67 theme RG-II 1526:1530 arg1 A 1542:1542 RG-II sidechain A 1526:1542 RG-II sidechain A 1526:1542 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 0 68 theme rhamnogalacturonan 96:113 arg1 structure 118:126 rhamnogalacturonan II structure 96:126 rhamnogalacturonan II structure 96:126 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 5 69 theme less 975:978 arg1 apiose 980:985 63 and 88% less apiose 964:985 63 and 88% less apiose 964:985 Here, we show that apiogalacturonan decreased substantially as the Lemnoideae diversified since Wolffiella and Wolffia walls contain between 63 and 88% less apiose than Spirodela, Landoltia, and Lemna walls. 29288327 7 70 from plants 1433:1438 arg1 RG-II 1410:1414 RG-II 1410:1414 RG-II from terrestrial plants 1410:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 8 71 theme A 1542:1542 arg1 methyl-etherification 1501:1521 methyl-etherification 1501:1521 methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B 1501:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 4 72 theme members 769:775 arg1 walls 754:758 the cell walls 745:758 the cell walls of other members of the Lemnoideae 745:793 However, the pectins in the cell walls of other members of the Lemnoideae have not been investigated. 29288327 0 73 theme structure 118:126 arg1 conservation 80:91 conservation 80:91 conservation of rhamnogalacturonan II structure 80:126 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 0 73 theme structure 118:126 arg1 xylogalacturonan 59:74 xylogalacturonan 59:74 xylogalacturonan 59:74 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 7 74 theme Lemnoideae 1323:1332 arg1 walls 1334:1338 Lemnoideae walls 1323:1338 Lemnoideae walls 1323:1338 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 8 75 theme arabinosylation 1548:1562 arg1 methyl-etherification 1501:1521 methyl-etherification 1501:1521 methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B 1501:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 2 76 theme small 470:474 arg1 monocots 498:505 small, fast-growing aquatic monocots 470:505 small, fast-growing aquatic monocots 470:505 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 8 77 theme sidechain 1567:1575 arg1 B 1577:1577 sidechain B 1567:1577 sidechain B 1567:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 4 78 theme cell 749:752 arg1 walls 754:758 the cell walls 745:758 the cell walls of other members of the Lemnoideae 745:793 However, the pectins in the cell walls of other members of the Lemnoideae have not been investigated. 29288327 9 79 theme Immunocytochemical 1596:1613 arg1 studies 1615:1621 Immunocytochemical studies 1596:1621 Immunocytochemical studies 1596:1621 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 7 80 link cross-linked 1352:1363 arg1 sequence 1390:1397 a glycosyl sequence 1379:1397 a glycosyl sequence similar to RG-II from terrestrial plants 1379:1438 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 80 link cross-linked 1352:1363 arg1 dimer 1365:1369 a borate cross-linked dimer 1343:1369 a borate cross-linked dimer 1343:1369 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 3 81 contain contain 602:608 arg1 walls 591:595 primary cell walls 578:595 primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants 578:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 81 contain contain 602:608 arg2 apiogalacturonan 627:642 apiogalacturonan 627:642 apiogalacturonan 627:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 81 contain contain 602:608 arg2 amounts 616:622 large amounts 610:622 large amounts of apiogalacturonan 610:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 82 theme primary 675:681 arg1 walls 683:687 the primary walls 671:687 the primary walls of most other flowering plants 671:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 0 83 theme cell 28:31 arg1 wall 33:36 cell wall 28:36 cell wall 28:36 Changes in the abundance of cell wall apiogalacturonan and xylogalacturonan and conservation of rhamnogalacturonan II structure during the diversification of the Lemnoideae. 29288327 2 84 theme monocots 498:505 arg1 genera 445:450 five genera 440:450 five genera 440:450 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 2 84 theme monocots 498:505 arg1 species 459:465 38 species 456:465 38 species 456:465 The subfamily Lemnoideae is comprised of five genera and 38 species of small, fast-growing aquatic monocots. 29288327 8 85 from variations 1473:1482 arg1 extent 1491:1496 the extent 1487:1496 the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B 1487:1577 Nevertheless, species-dependent variations in the extent of methyl-etherification of RG-II sidechain A and arabinosylation of sidechain B are discernible. 29288327 9 86 theme pectin 1637:1642 arg1 methyl-esterification 1644:1664 pectin methyl-esterification 1637:1664 pectin methyl-esterification 1637:1664 Immunocytochemical studies revealed that pectin methyl-esterification is higher in developing daughter frond walls than in mother frond walls, indicating that methyl-esterification is associated with expanding cells. 29288327 7 87 theme pectin 1276:1281 arg1 RG-II 1306:1310 RG-II 1306:1310 RG-II 1306:1310 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 7 87 theme pectin 1276:1281 arg1 rhamnogalacturonan 1283:1300 The apiose-containing pectin rhamnogalacturonan II 1254:1303 The apiose-containing pectin rhamnogalacturonan II (RG-II) 1254:1311 The apiose-containing pectin rhamnogalacturonan II (RG-II) exists in Lemnoideae walls as a borate cross-linked dimer and has a glycosyl sequence similar to RG-II from terrestrial plants. 29288327 3 88 theme other 697:701 arg1 plants 713:718 most other flowering plants 692:718 most other flowering plants 692:718 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 89 theme apiogalacturonan 627:642 arg1 apiogalacturonan 627:642 apiogalacturonan 627:642 apiogalacturonan 627:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 3 89 theme apiogalacturonan 627:642 arg1 amounts 616:622 large amounts 610:622 large amounts of apiogalacturonan 610:642 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 6 90 contain have 1173:1176 arg2 content 1195:1201 a high arabinose content 1178:1201 a high arabinose content 1178:1201 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 6 90 contain have 1173:1176 arg1 polysaccharides 1157:1171 in Wolffiella pectic polysaccharides 1136:1171 in Wolffiella pectic polysaccharides 1136:1171 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 29288327 3 91 theme minor 514:518 arg1 Lemna 508:512 Lemna minor 508:518 Lemna minor 508:518 Lemna minor and Spirodela polyrhiza belong to this subfamily and have primary cell walls that contain large amounts of apiogalacturonan and thus are distinct from the primary walls of most other flowering plants. 29288327 4 92 from pectins 734:740 arg1 walls 754:758 the cell walls 745:758 the cell walls of other members of the Lemnoideae 745:793 However, the pectins in the cell walls of other members of the Lemnoideae have not been investigated. 29288327 6 93 from abundant 1096:1103 arg1 Wolffia 1034:1040 Wolffia 1034:1040 Wolffia 1034:1040 In Wolffia, the most derived genus, xylogalacturonan is far more abundant than apiogalacturonan, whereas in Wolffiella pectic polysaccharides have a high arabinose content, which may arise from arabinan sidechains of RG I. 24341799 3 0 theme reaction 709:716 arg1 coordinate 718:727 the 'dynamic' reaction coordinate 695:727 the 'dynamic' reaction coordinate for the catalytic cycle 695:751 We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. 24341799 1 1 theme mechanisms 253:262 arg1 latter 269:274 latter 269:274 latter 269:274 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 2 2 with structures 397:406 arg1 ligand 542:547 a full cellononaose ligand 522:547 a full cellononaose ligand 522:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 5 3 theme intermediate 1092:1103 arg1 carbon 1062:1067 the anomeric carbon 1049:1067 the anomeric carbon of the glycosyl-enzyme intermediate 1049:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 5 4 theme water 1007:1011 arg1 molecule 1013:1020 a water molecule 1005:1020 a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate 1005:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 0 5 theme structural 95:104 arg1 studies 135:141 structural and transition path sampling studies 95:141 structural and transition path sampling studies 95:141 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 4 6 from change 863:868 arg1 nucleophile 877:887 the nucleophile 873:887 the nucleophile 873:887 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 1 7 theme glycosyl-enzyme 348:362 arg1 intermediate 364:375 a glycosyl-enzyme intermediate 346:375 a glycosyl-enzyme intermediate 346:375 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 6 8 theme naturally 1223:1231 arg1 family 1267:1272 this naturally and industrially important enzyme family 1218:1272 this naturally and industrially important enzyme family 1218:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 1 9 theme intermediate 364:375 arg1 formation 333:341 formation 333:341 formation of a glycosyl-enzyme intermediate 333:375 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 2 10 theme reaction 621:628 arg1 coordinate 630:639 the 'static' reaction coordinate 608:639 the 'static' reaction coordinate 608:639 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 11 theme coordinate 630:639 arg1 details 597:603 details 597:603 details of the 'static' reaction coordinate 597:639 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 4 12 theme reaction 772:779 arg1 coordinate 781:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 5 13 from attack 1039:1044 arg1 carbon 1062:1067 the anomeric carbon 1049:1067 the anomeric carbon of the glycosyl-enzyme intermediate 1049:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 3 14 theme dynamic 700:706 arg1 coordinate 718:727 the 'dynamic' reaction coordinate 695:727 the 'dynamic' reaction coordinate for the catalytic cycle 695:751 We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. 24341799 5 15 dep mechanism 938:946 arg1 wherein 948:954 wherein 948:954 wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate 948:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 4 16 theme glycosylation 758:770 arg1 coordinate 781:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 6 17 theme previous 1122:1129 arg1 structures 1131:1140 previous structures 1122:1140 previous structures 1122:1140 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 2 18 theme new 393:395 arg1 structures 397:406 two new structures 389:406 two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 389:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 19 theme cellobiohydrolase 465:481 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 20 theme static 613:618 arg1 coordinate 630:639 the 'static' reaction coordinate 608:639 the 'static' reaction coordinate 608:639 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 21 with complex 509:515 arg1 ligand 542:547 a full cellononaose ligand 522:547 a full cellononaose ligand 522:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 5 22 theme glycosylation 960:972 arg1 cellobiose 983:992 cellobiose 983:992 cellobiose 983:992 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 5 22 theme glycosylation 960:972 arg1 product 974:980 the glycosylation product 956:980 the glycosylation product 956:980 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 1 23 theme glycosidic 178:187 arg1 linkages 189:196 glycosidic linkages 178:196 glycosidic linkages in carbohydrates 178:213 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 2 24 theme jecorina 444:451 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 3 25 theme catalytic 737:745 arg1 cycle 747:751 the catalytic cycle 733:751 the catalytic cycle 733:751 We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. 24341799 2 26 theme full 524:527 arg1 ligand 542:547 a full cellononaose ligand 522:547 a full cellononaose ligand 522:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 6 27 theme reaction 1194:1201 arg1 coordinate 1203:1212 the complete hydrolytic reaction coordinate 1170:1212 the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family 1170:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 0 28 theme cellulose 17:25 arg1 hydrolysis 27:36 cellulose hydrolysis 17:36 cellulose hydrolysis 17:36 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 6 29 theme important 1250:1258 arg1 family 1267:1272 this naturally and industrially important enzyme family 1218:1272 this naturally and industrially important enzyme family 1218:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 0 30 theme sampling 126:133 arg1 studies 135:141 structural and transition path sampling studies 95:141 structural and transition path sampling studies 95:141 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 6 31 theme hydrolytic 1183:1192 arg1 coordinate 1203:1212 the complete hydrolytic reaction coordinate 1170:1212 the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family 1170:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 2 32 theme domain 425:430 arg1 intermediate 571:582 a glycosyl-enzyme intermediate 553:582 a glycosyl-enzyme intermediate 553:582 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 32 theme domain 425:430 arg1 structures 397:406 two new structures 389:406 two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 389:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 32 theme domain 425:430 arg1 complex 509:515 namely a Michaelis complex 490:515 namely a Michaelis complex with a full cellononaose ligand 490:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 6 33 theme complete 1174:1181 arg1 coordinate 1203:1212 the complete hydrolytic reaction coordinate 1170:1212 the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family 1170:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 2 34 theme catalytic 415:423 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 34 theme catalytic 415:423 arg1 domain 425:430 the catalytic domain 411:430 the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 411:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 35 theme Cel7A 483:487 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 35 theme Cel7A 483:487 arg1 domain 425:430 the catalytic domain 411:430 the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 411:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 0 36 dep two-step 43:50 arg1 retaining 53:61 retaining 53:61 retaining 53:61 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 0 37 theme hydrolysis 27:36 arg1 mechanism 4:12 The mechanism 0:12 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase 0:79 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 2 38 theme cellononaose 529:540 arg1 ligand 542:547 a full cellononaose ligand 522:547 a full cellononaose ligand 522:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 39 theme Hypocrea 435:442 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 4 40 from bonds 836:840 arg1 nucleophile 877:887 the nucleophile 873:887 the nucleophile 873:887 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 0 41 theme two-step 43:50 arg1 cellobiohydrolase 63:79 a two-step, retaining cellobiohydrolase 41:79 a two-step, retaining cellobiohydrolase 41:79 The mechanism of cellulose hydrolysis by a two-step, retaining cellobiohydrolase elucidated by structural and transition path sampling studies. 24341799 1 42 theme two-step 300:307 arg1 mechanism 309:317 a two-step mechanism 298:317 a two-step mechanism that includes formation of a glycosyl-enzyme intermediate 298:375 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 6 43 with concert 1109:1115 arg1 structures 1131:1140 previous structures 1122:1140 previous structures 1122:1140 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 5 44 theme anomeric 1053:1060 arg1 carbon 1062:1067 the anomeric carbon 1049:1067 the anomeric carbon of the glycosyl-enzyme intermediate 1049:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 4 45 theme conformational 848:861 arg1 change 863:868 a conformational change 846:868 a conformational change in the nucleophile 846:887 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 5 46 theme product-assisted 921:936 arg1 mechanism 938:946 a product-assisted mechanism 919:946 a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate 919:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 2 47 theme Family 456:461 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 3 48 theme path 668:671 arg1 sampling 673:680 transition path sampling 657:680 transition path sampling 657:680 We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. 24341799 4 49 theme breaking 827:834 arg1 bonds 836:840 breaking bonds 827:840 breaking bonds 827:840 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 2 50 theme glycosyl-enzyme 555:569 arg1 intermediate 571:582 a glycosyl-enzyme intermediate 553:582 a glycosyl-enzyme intermediate 553:582 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 5 51 theme glycosyl-enzyme 1076:1090 arg1 intermediate 1092:1103 the glycosyl-enzyme intermediate 1072:1103 the glycosyl-enzyme intermediate 1072:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 6 52 theme enzyme 1260:1265 arg1 family 1267:1272 this naturally and industrially important enzyme family 1218:1272 this naturally and industrially important enzyme family 1218:1272 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 6 53 theme present 1147:1153 arg1 results 1155:1161 the present results 1143:1161 the present results 1143:1161 In concert with previous structures, the present results reveal the complete hydrolytic reaction coordinate for this naturally and industrially important enzyme family. 24341799 1 54 theme inverting 230:238 arg1 mechanisms 253:262 inverting or retaining mechanisms 230:262 inverting or retaining mechanisms 230:262 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 2 55 theme Michaelis 499:507 arg1 complex 509:515 namely a Michaelis complex 490:515 namely a Michaelis complex with a full cellononaose ligand 490:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 3 56 theme transition 657:666 arg1 sampling 673:680 transition path sampling 657:680 transition path sampling 657:680 We also employ transition path sampling to determine the 'dynamic' reaction coordinate for the catalytic cycle. 24341799 5 57 theme nucleophilic 1026:1037 arg1 attack 1039:1044 nucleophilic attack 1026:1044 nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate 1026:1103 Deglycosylation proceeds via a product-assisted mechanism wherein the glycosylation product, cellobiose, positions a water molecule for nucleophilic attack on the anomeric carbon of the glycosyl-enzyme intermediate. 24341799 1 58 from linkages 189:196 arg1 carbohydrates 201:213 carbohydrates 201:213 carbohydrates 201:213 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 1 59 theme Glycoside 144:152 arg1 hydrolases 154:163 Glycoside hydrolases 144:163 Glycoside hydrolases (GHs) 144:169 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 1 59 theme Glycoside 144:152 arg1 GHs 166:168 GHs 166:168 GHs 166:168 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 24341799 4 60 contain contains 792:799 arg1 coordinate 781:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate 754:790 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 4 60 contain contains 792:799 arg2 components 801:810 components 801:810 components of forming and breaking bonds and a conformational change in the nucleophile 801:887 The glycosylation reaction coordinate contains components of forming and breaking bonds and a conformational change in the nucleophile. 24341799 2 61 theme GH 453:454 arg1 Cel7A 483:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A 435:487 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 2 62 with intermediate 571:582 arg1 ligand 542:547 a full cellononaose ligand 522:547 a full cellononaose ligand 522:547 We present two new structures of the catalytic domain of Hypocrea jecorina GH Family 7 cellobiohydrolase Cel7A, namely a Michaelis complex with a full cellononaose ligand and a glycosyl-enzyme intermediate, that reveal details of the 'static' reaction coordinate. 24341799 1 63 theme retaining 243:251 arg1 mechanisms 253:262 inverting or retaining mechanisms 230:262 inverting or retaining mechanisms 230:262 Glycoside hydrolases (GHs) cleave glycosidic linkages in carbohydrates, typically via inverting or retaining mechanisms, the latter of which proceeds via a two-step mechanism that includes formation of a glycosyl-enzyme intermediate. 29062024 4 0 gly fucosylation 784:795 arg1 IgG1-Fc 800:806 IgG1-Fc 800:806 IgG1-Fc 800:806 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 6 1 theme core 1359:1362 arg1 fucosylation 1364:1375 Fc core fucosylation 1356:1375 Fc core fucosylation 1356:1375 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 4 2 theme increased 703:711 arg1 fluctuation 728:738 increased conformational fluctuation 703:738 increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa 703:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 0 3 theme region 76:81 arg1 fucosylation 40:51 N-glycan core fucosylation 26:51 N-glycan core fucosylation of immunoglobulin G Fc region 26:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 0 4 from effects 15:21 arg1 interaction 90:100 its interaction 86:100 its interaction with Fcγ receptor IIIa 86:123 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 6 5 theme Fc 1356:1357 arg1 fucosylation 1364:1375 Fc core fucosylation 1356:1375 Fc core fucosylation 1356:1375 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 4 6 theme IgG1-Fc 800:806 arg1 fucosylation 784:795 fucosylation 784:795 fucosylation of IgG1-Fc 784:806 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 4 7 with consistent 809:818 arg1 data 842:845 crystallographic data 825:845 crystallographic data giving no interpretable electron density for this N-glycan 825:904 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 6 8 theme IgG1-Fc 1152:1158 arg1 fucosylation 1136:1147 core fucosylation 1131:1147 core fucosylation of IgG1-Fc 1131:1158 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 3 9 theme FcγRIIIa 631:638 arg1 form 623:626 a soluble form 613:626 a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 613:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 7 10 theme therapeutic 1450:1460 arg1 antibodies 1462:1471 therapeutic antibodies 1450:1471 therapeutic antibodies with improved ADCC activity 1450:1499 Our findings offer a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity. 29062024 3 11 theme replica-exchange 527:542 arg1 simulations 563:573 replica-exchange molecular dynamics simulations 527:573 replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 527:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 5 12 theme carbohydrate-carbohydrate 989:1013 arg1 interactions 1015:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 5 13 theme optimum 966:972 arg1 interactions 1015:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 1 14 theme glycoproteins 328:340 arg1 N-glycosylation 303:317 N-glycosylation 303:317 N-glycosylation of these glycoproteins 303:340 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 4 15 theme sFcγRIIIa 769:777 arg1 Asn162 759:764 the N-glycan at Asn162 743:764 the N-glycan at Asn162 of sFcγRIIIa 743:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 3 16 theme molecular 544:552 arg1 simulations 563:573 replica-exchange molecular dynamics simulations 527:573 replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 527:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 1 17 gly N-glycosylation 303:317 arg1 glycoproteins 328:340 these glycoproteins 322:340 these glycoproteins 322:340 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 0 18 theme Fcγ 107:109 arg1 IIIa 120:123 Fcγ receptor IIIa 107:123 Fcγ receptor IIIa 107:123 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 3 19 with complex 655:661 arg1 IgG1-Fc 668:674 IgG1-Fc 668:674 IgG1-Fc 668:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 2 20 gly fucosylation 363:374 arg1 N-glycans 387:395 IgG1-Fc N-glycans 379:395 IgG1-Fc N-glycans 379:395 In particular, core fucosylation of IgG1-Fc N-glycans negatively affects this interaction and thereby compromises ADCC activity. 29062024 3 21 from form 623:626 arg1 complex 655:661 complex 655:661 complex with IgG1-Fc 655:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 7 22 theme structural 1399:1408 arg1 foundation 1410:1419 a structural foundation 1397:1419 a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity 1397:1499 Our findings offer a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity. 29062024 4 23 theme interpretable 857:869 arg1 density 880:886 no interpretable electron density 854:886 no interpretable electron density 854:886 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 6 24 with interaction 1321:1331 arg1 sFcγRIIIa 1338:1346 sFcγRIIIa 1338:1346 sFcγRIIIa 1338:1346 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 6 25 theme core 1131:1134 arg1 fucosylation 1136:1147 core fucosylation 1131:1147 core fucosylation of IgG1-Fc 1131:1158 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 6 26 theme residues 1237:1244 arg1 rearrangements 1196:1209 rearrangements 1196:1209 rearrangements 1196:1209 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 6 26 theme residues 1237:1244 arg1 dynamics 1183:1190 conformational dynamics 1168:1190 conformational dynamics 1168:1190 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 3 27 from complex 655:661 arg1 form 623:626 a soluble form 613:626 a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 613:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 4 28 theme at 756:757 arg1 Asn162 759:764 the N-glycan at Asn162 743:764 the N-glycan at Asn162 of sFcγRIIIa 743:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 3 29 from FcγRIIIa 631:638 arg1 complex 655:661 complex 655:661 complex with IgG1-Fc 655:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 2 30 theme core 358:361 arg1 fucosylation 363:374 core fucosylation 358:374 core fucosylation of IgG1-Fc N-glycans 358:395 In particular, core fucosylation of IgG1-Fc N-glycans negatively affects this interaction and thereby compromises ADCC activity. 29062024 0 31 theme Conformational 0:13 arg1 effects 15:21 Conformational effects 0:21 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 0:124 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 1 32 theme Fcγ 260:262 arg1 IIIa 273:276 Fcγ receptor IIIa 260:276 Fcγ receptor IIIa (FcγRIIIa) 260:287 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 32 theme Fcγ 260:262 arg1 FcγRIIIa 279:286 FcγRIIIa 279:286 FcγRIIIa 279:286 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 0 33 with interaction 90:100 arg1 IIIa 120:123 Fcγ receptor IIIa 107:123 Fcγ receptor IIIa 107:123 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 4 34 theme electron 871:878 arg1 density 880:886 no interpretable electron density 854:886 no interpretable electron density 854:886 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 5 35 from glycan 1080:1085 arg1 distal 1061:1066 distal 1061:1066 distal 1061:1066 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 6 36 theme IgG1-Fc 1269:1275 arg1 Tyr296 1259:1264 Tyr296 1259:1264 Tyr296 of IgG1-Fc 1259:1275 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 0 37 theme N-glycan 26:33 arg1 fucosylation 40:51 N-glycan core fucosylation 26:51 N-glycan core fucosylation of immunoglobulin G Fc region 26:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 1 38 theme receptor 264:271 arg1 IIIa 273:276 Fcγ receptor IIIa 260:276 Fcγ receptor IIIa (FcγRIIIa) 260:287 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 38 theme receptor 264:271 arg1 FcγRIIIa 279:286 FcγRIIIa 279:286 FcγRIIIa 279:286 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 3 39 theme dynamics 554:561 arg1 simulations 563:573 replica-exchange molecular dynamics simulations 527:573 replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 527:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 1 40 gly glycoproteins 328:340 arg1 glycoproteins 328:340 these glycoproteins 322:340 these glycoproteins 322:340 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 4 41 theme N-glycan 747:754 arg1 Asn162 759:764 the N-glycan at Asn162 743:764 the N-glycan at Asn162 of sFcγRIIIa 743:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 1 42 theme IIIa 273:276 arg1 region 221:226 the Fc region 214:226 the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa) 214:287 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 0 43 theme receptor 111:118 arg1 IIIa 120:123 Fcγ receptor IIIa 107:123 Fcγ receptor IIIa 107:123 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 4 44 theme Asn162 759:764 arg1 fluctuation 728:738 increased conformational fluctuation 703:738 increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa 703:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 2 45 theme ADCC 457:460 arg1 activity 462:469 ADCC activity 457:469 ADCC activity 457:469 In particular, core fucosylation of IgG1-Fc N-glycans negatively affects this interaction and thereby compromises ADCC activity. 29062024 3 46 theme effect 506:511 arg1 mechanisms 487:496 the mechanisms 483:496 the mechanisms of this effect 483:511 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 0 47 theme fucosylation 40:51 arg1 effects 15:21 Conformational effects 0:21 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 0:124 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 2 48 theme N-glycans 387:395 arg1 fucosylation 363:374 core fucosylation 358:374 core fucosylation of IgG1-Fc N-glycans 358:395 In particular, core fucosylation of IgG1-Fc N-glycans negatively affects this interaction and thereby compromises ADCC activity. 29062024 0 49 theme core 35:38 arg1 fucosylation 40:51 N-glycan core fucosylation 26:51 N-glycan core fucosylation of immunoglobulin G Fc region 26:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 7 50 theme ADCC 1487:1490 arg1 activity 1492:1499 improved ADCC activity 1478:1499 improved ADCC activity 1478:1499 Our findings offer a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity. 29062024 4 51 theme crystallographic 825:840 arg1 data 842:845 crystallographic data 825:845 crystallographic data giving no interpretable electron density for this N-glycan 825:904 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 1 52 theme Fc 218:219 arg1 region 221:226 the Fc region 214:226 the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa) 214:287 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 0 53 theme immunoglobulin 56:69 arg1 region 76:81 immunoglobulin G Fc region 56:81 immunoglobulin G Fc region 56:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 7 54 theme improved 1478:1485 arg1 activity 1492:1499 improved ADCC activity 1478:1499 improved ADCC activity 1478:1499 Our findings offer a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity. 29062024 3 55 theme crystallographic 584:599 arg1 analysis 601:608 crystallographic analysis 584:608 crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 584:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 6 56 theme conformational 1168:1181 arg1 dynamics 1183:1190 conformational dynamics 1168:1190 conformational dynamics 1168:1190 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 5 57 theme intermolecular 974:987 arg1 interactions 1015:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 optimum intermolecular carbohydrate-carbohydrate interactions 966:1026 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 6 58 gly fucosylation 1136:1147 arg1 residues 1237:1244 surrounding amino acid residues 1214:1244 surrounding amino acid residues 1214:1244 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 6 58 gly fucosylation 1136:1147 arg1 IgG1-Fc 1152:1158 IgG1-Fc 1152:1158 IgG1-Fc 1152:1158 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 6 59 theme amino 1226:1230 arg1 residues 1237:1244 surrounding amino acid residues 1214:1244 surrounding amino acid residues 1214:1244 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 4 60 theme conformational 713:726 arg1 fluctuation 728:738 increased conformational fluctuation 703:738 increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa 703:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 5 61 theme sFcγRIIIa 1044:1052 arg1 glycan 1054:1059 this sFcγRIIIa glycan 1039:1059 this sFcγRIIIa glycan 1039:1059 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 5 62 theme Fc 1077:1078 arg1 glycan 1080:1085 the Fc glycan 1073:1085 the Fc glycan 1073:1085 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 5 63 theme fucose 942:947 arg1 residue 949:955 The fucose residue 938:955 The fucose residue 938:955 The fucose residue disrupts optimum intermolecular carbohydrate-carbohydrate interactions, rendering this sFcγRIIIa glycan distal from the Fc glycan. 29062024 4 64 from Asn162 759:764 arg1 at 756:757 the N-glycan at Asn162 743:764 the N-glycan at Asn162 of sFcγRIIIa 743:777 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 4 65 theme innermost 922:930 arg1 part 932:935 the innermost part 918:935 the innermost part 918:935 Our simulation highlights increased conformational fluctuation of the N-glycan at Asn162 of sFcγRIIIa upon fucosylation of IgG1-Fc, consistent with crystallographic data giving no interpretable electron density for this N-glycan, except for the innermost part. 29062024 0 66 gly fucosylation 40:51 arg1 region 76:81 immunoglobulin G Fc region 56:81 immunoglobulin G Fc region 56:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 1 67 theme Antibody-dependent 126:143 arg1 cytotoxicity 154:165 Antibody-dependent cellular cytotoxicity 126:165 Antibody-dependent cellular cytotoxicity (ADCC) 126:172 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 67 theme Antibody-dependent 126:143 arg1 ADCC 168:171 ADCC 168:171 ADCC 168:171 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 7 68 with antibodies 1462:1471 arg1 activity 1492:1499 improved ADCC activity 1478:1499 improved ADCC activity 1478:1499 Our findings offer a structural foundation for designing and developing therapeutic antibodies with improved ADCC activity. 29062024 0 69 theme Fc 73:74 arg1 region 76:81 immunoglobulin G Fc region 56:81 immunoglobulin G Fc region 56:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 2 70 theme IgG1-Fc 379:385 arg1 N-glycans 387:395 IgG1-Fc N-glycans 379:395 IgG1-Fc N-glycans 379:395 In particular, core fucosylation of IgG1-Fc N-glycans negatively affects this interaction and thereby compromises ADCC activity. 29062024 3 71 theme soluble 615:621 arg1 form 623:626 a soluble form 613:626 a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 613:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 1 72 theme cellular 145:152 arg1 cytotoxicity 154:165 Antibody-dependent cellular cytotoxicity 126:165 Antibody-dependent cellular cytotoxicity (ADCC) 126:172 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 72 theme cellular 145:152 arg1 ADCC 168:171 ADCC 168:171 ADCC 168:171 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 73 theme immunoglobulin 231:244 arg1 IgG1 250:253 IgG1 250:253 IgG1 250:253 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 1 73 theme immunoglobulin 231:244 arg1 G1 246:247 immunoglobulin G1 231:247 immunoglobulin G1 (IgG1) 231:254 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 29062024 0 74 theme G 71:71 arg1 region 76:81 immunoglobulin G Fc region 56:81 immunoglobulin G Fc region 56:81 Conformational effects of N-glycan core fucosylation of immunoglobulin G Fc region on its interaction with Fcγ receptor IIIa. 29062024 6 75 theme surrounding 1214:1224 arg1 residues 1237:1244 surrounding amino acid residues 1214:1244 surrounding amino acid residues 1214:1244 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 3 76 theme form 623:626 arg1 analysis 601:608 crystallographic analysis 584:608 crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc 584:674 To address the mechanisms of this effect, we performed replica-exchange molecular dynamics simulations based on crystallographic analysis of a soluble form of FcγRIIIa (sFcγRIIIa) in complex with IgG1-Fc. 29062024 6 77 theme acid 1232:1235 arg1 residues 1237:1244 surrounding amino acid residues 1214:1244 surrounding amino acid residues 1214:1244 Moreover, our simulation demonstrates that core fucosylation of IgG1-Fc affects conformational dynamics and rearrangements of surrounding amino acid residues, typified by Tyr296 of IgG1-Fc, which was more extensively involved in the interaction with sFcγRIIIa without Fc core fucosylation. 29062024 1 78 theme G1 246:247 arg1 region 221:226 the Fc region 214:226 the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa) 214:287 Antibody-dependent cellular cytotoxicity (ADCC) is promoted through interaction between the Fc region of immunoglobulin G1 (IgG1) and Fcγ receptor IIIa (FcγRIIIa), depending on N-glycosylation of these glycoproteins. 27333379 6 0 from IgA1 1007:1010 arg1 generation 959:968 the generation 955:968 the generation of distinct O-glycans on recombinant IgA1 955:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 6 1 theme recombinant 995:1005 arg1 IgA1 1007:1010 recombinant IgA1 995:1010 recombinant IgA1 995:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 5 2 theme other 709:713 arg1 hosts 726:730 other expression hosts 709:730 other expression hosts 709:730 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 1 3 theme autoimmune 138:147 arg1 nephropathy 107:117 IgA nephropathy 103:117 IgA nephropathy (IgAN) 103:124 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 1 3 theme autoimmune 138:147 arg1 disease 149:155 a common autoimmune disease 129:155 a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease 129:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 4 4 from IgA1 618:621 arg1 overproduction 578:591 the overproduction 574:591 the overproduction of aberrant O-glycans on IgA1 574:621 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 5 5 theme de 766:767 arg1 synthesis 774:782 de novo synthesis 766:782 de novo synthesis of O-glycans 766:795 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 4 6 from overproduction 578:591 arg1 IgA1 618:621 IgA1 618:621 IgA1 618:621 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 5 7 theme expression 715:724 arg1 hosts 726:730 other expression hosts 709:730 other expression hosts 709:730 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 7 8 theme engineering 1033:1043 arg1 repertoire 1045:1054 our engineering repertoire 1029:1054 our engineering repertoire 1029:1054 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 3 9 theme circulating 372:382 arg1 IgA1 384:387 circulating IgA1 372:387 circulating IgA1 372:387 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 6 10 theme O-glycans 982:990 arg1 generation 959:968 the generation 955:968 the generation of distinct O-glycans on recombinant IgA1 955:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 1 11 theme end-stage 265:273 arg1 disease 282:288 end-stage kidney disease 265:288 end-stage kidney disease 265:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 5 12 theme O-glycans 787:795 arg1 synthesis 774:782 de novo synthesis 766:782 de novo synthesis of O-glycans 766:795 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 6 13 theme distinct 973:980 arg1 O-glycans 982:990 distinct O-glycans 973:990 distinct O-glycans on recombinant IgA1 973:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 5 14 gly glycoproteins 812:824 arg1 glycoproteins 812:824 recombinant glycoproteins 800:824 recombinant glycoproteins 800:824 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 1 15 theme kidney 275:280 arg1 disease 282:288 end-stage kidney disease 265:288 end-stage kidney disease 265:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 5 16 theme O-glycosylation 860:874 arg1 pathway 876:882 the mammalian O-glycosylation pathway 846:882 the mammalian O-glycosylation pathway 846:882 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 4 17 theme specific 624:631 arg1 structure-function-studies 633:658 specific structure-function-studies 624:658 specific structure-function-studies of mucin-type O-glycans 624:682 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 3 18 dep bound 440:444 arg1 contribute 474:483 contribute 474:483 contribute to immune complex formation and pathogenesis 474:528 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 3 19 theme mucin-type 415:424 arg1 O-glycans 426:434 these galactose-deficient mucin-type O-glycans 389:434 these galactose-deficient mucin-type O-glycans 389:434 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 5 20 dep de 766:767 arg1 novo 769:772 novo 769:772 novo 769:772 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 4 21 from O-glycans 605:613 arg1 IgA1 618:621 IgA1 618:621 IgA1 618:621 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 4 22 theme aberrant 596:603 arg1 O-glycans 605:613 aberrant O-glycans 596:613 aberrant O-glycans on IgA1 596:621 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 2 23 theme immune 309:314 arg1 complexes 316:324 these immune complexes 303:324 these immune complexes 303:324 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 1 24 theme IgA 103:105 arg1 disease 149:155 a common autoimmune disease 129:155 a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease 129:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 1 24 theme IgA 103:105 arg1 IgAN 120:123 IgAN 120:123 IgAN 120:123 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 1 24 theme IgA 103:105 arg1 nephropathy 107:117 IgA nephropathy 103:117 IgA nephropathy (IgAN) 103:124 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 5 25 theme recombinant 800:810 arg1 glycoproteins 812:824 recombinant glycoproteins 800:824 recombinant glycoproteins 800:824 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 3 26 theme complex 495:501 arg1 formation 503:511 immune complex formation 488:511 immune complex formation 488:511 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 3 27 theme galactose-deficient 395:413 arg1 O-glycans 426:434 these galactose-deficient mucin-type O-glycans 389:434 these galactose-deficient mucin-type O-glycans 389:434 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 7 28 from patients 1188:1195 arg1 IgA1 1178:1181 IgA1 1178:1181 IgA1 from patients with IgAN 1178:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 4 29 theme mucin-type 663:672 arg1 O-glycans 674:682 mucin-type O-glycans 663:682 mucin-type O-glycans 663:682 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 0 30 theme recombinant 40:50 arg1 IgA1 52:55 recombinant IgA1 40:55 recombinant IgA1 40:55 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 4 31 theme O-glycans 605:613 arg1 overproduction 578:591 the overproduction 574:591 the overproduction of aberrant O-glycans on IgA1 574:621 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 27333379 7 32 theme prevailing 1147:1156 arg1 glycans 1158:1164 the prevailing glycans 1143:1164 the prevailing glycans detected on IgA1 from patients with IgAN 1143:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 32 theme prevailing 1147:1156 arg1 O-glycans 1123:1131 galactose-deficient and α2,6-sialylated O-glycans 1083:1131 galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1083:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 0 33 from O-glycans 82:90 arg1 plants 95:100 plants 95:100 plants 95:100 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 7 34 gly α2,6-sialylated 1107:1121 arg1 glycans 1158:1164 the prevailing glycans 1143:1164 the prevailing glycans detected on IgA1 from patients with IgAN 1143:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 34 gly α2,6-sialylated 1107:1121 arg1 O-glycans 1123:1131 galactose-deficient and α2,6-sialylated O-glycans 1083:1131 galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1083:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 0 35 with production 26:35 arg1 O-glycans 82:90 distinct mucin-type O-glycans 62:90 distinct mucin-type O-glycans in plants 62:100 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 3 36 theme immune 488:493 arg1 formation 503:511 immune complex formation 488:511 immune complex formation 488:511 In circulating IgA1 these galactose-deficient mucin-type O-glycans are bound by autoantibodies and thus, contribute to immune complex formation and pathogenesis. 27333379 5 37 theme mammalian 850:858 arg1 pathway 876:882 the mammalian O-glycosylation pathway 846:882 the mammalian O-glycosylation pathway 846:882 Compared to other expression hosts, plants offer the opportunity for de novo synthesis of O-glycans on recombinant glycoproteins as they are lacking the mammalian O-glycosylation pathway. 27333379 6 38 from generation 959:968 arg1 IgA1 1007:1010 recombinant IgA1 995:1010 recombinant IgA1 995:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 2 39 gly glycosylated 345:356 arg1 O-glycans 358:366 aberrantly glycosylated O-glycans 334:366 aberrantly glycosylated O-glycans 334:366 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 6 40 from O-glycans 982:990 arg1 IgA1 1007:1010 recombinant IgA1 995:1010 recombinant IgA1 995:1010 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 7 41 theme α2,6-sialylated 1107:1121 arg1 glycans 1158:1164 the prevailing glycans 1143:1164 the prevailing glycans detected on IgA1 from patients with IgAN 1143:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 41 theme α2,6-sialylated 1107:1121 arg1 O-glycans 1123:1131 galactose-deficient and α2,6-sialylated O-glycans 1083:1131 galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1083:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 0 42 theme IgA1 52:55 arg1 production 26:35 the production 22:35 the production of recombinant IgA1 with distinct mucin-type O-glycans in plants 22:100 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 6 43 theme Nicotiana 916:924 arg1 benthamiana 926:936 Nicotiana benthamiana 916:936 Nicotiana benthamiana 916:936 Recently, we demonstrated that Nicotiana benthamiana are suitable for the generation of distinct O-glycans on recombinant IgA1. 27333379 2 44 theme glycosylated 345:356 arg1 O-glycans 358:366 aberrantly glycosylated O-glycans 334:366 aberrantly glycosylated O-glycans 334:366 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 7 45 dep expand 1022:1027 arg1 by 1056:1057 by 1056:1057 by 1056:1057 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 46 theme O-glycans 1123:1131 arg1 generation 1069:1078 planta generation 1062:1078 planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1062:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 47 theme galactose-deficient 1083:1101 arg1 glycans 1158:1164 the prevailing glycans 1143:1164 the prevailing glycans detected on IgA1 from patients with IgAN 1143:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 47 theme galactose-deficient 1083:1101 arg1 O-glycans 1123:1131 galactose-deficient and α2,6-sialylated O-glycans 1083:1131 galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1083:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 48 with patients 1188:1195 arg1 IgAN 1202:1205 IgAN 1202:1205 IgAN 1202:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 1 49 theme IgA1-containing 210:224 arg1 complexes 233:241 IgA1-containing immune complexes 210:241 IgA1-containing immune complexes frequently leading to end-stage kidney disease 210:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 7 50 theme planta 1062:1067 arg1 generation 1069:1078 planta generation 1062:1078 planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1062:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 51 located detected 1166:1173 arg2 O-glycans 1123:1131 galactose-deficient and α2,6-sialylated O-glycans 1083:1131 galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN 1083:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 51 located detected 1166:1173 arg1 IgA1 1178:1181 IgA1 1178:1181 IgA1 from patients with IgAN 1178:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 7 51 located detected 1166:1173 arg2 glycans 1158:1164 the prevailing glycans 1143:1164 the prevailing glycans detected on IgA1 from patients with IgAN 1143:1205 Here, we expand our engineering repertoire by in planta generation of galactose-deficient and α2,6-sialylated O-glycans which are the prevailing glycans detected on IgA1 from patients with IgAN. 27333379 2 52 from IgA1 295:298 arg1 complexes 316:324 these immune complexes 303:324 these immune complexes 303:324 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 1 53 theme immune 226:231 arg1 complexes 233:241 IgA1-containing immune complexes 210:241 IgA1-containing immune complexes frequently leading to end-stage kidney disease 210:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 0 54 theme mucin-type 71:80 arg1 O-glycans 82:90 distinct mucin-type O-glycans 62:90 distinct mucin-type O-glycans in plants 62:100 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 1 55 theme complexes 233:241 arg1 deposition 196:205 deposition 196:205 deposition 196:205 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 1 55 theme complexes 233:241 arg1 formation 182:190 formation 182:190 formation 182:190 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 0 56 theme distinct 62:69 arg1 O-glycans 82:90 distinct mucin-type O-glycans 62:90 distinct mucin-type O-glycans in plants 62:100 Glyco-engineering for the production of recombinant IgA1 with distinct mucin-type O-glycans in plants. 27333379 2 57 contain carries 326:332 arg1 IgA1 295:298 The IgA1 291:298 The IgA1 in these immune complexes 291:324 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 2 57 contain carries 326:332 arg2 O-glycans 358:366 aberrantly glycosylated O-glycans 334:366 aberrantly glycosylated O-glycans 334:366 The IgA1 in these immune complexes carries aberrantly glycosylated O-glycans. 27333379 1 58 theme common 131:136 arg1 nephropathy 107:117 IgA nephropathy 103:117 IgA nephropathy (IgAN) 103:124 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 1 58 theme common 131:136 arg1 disease 149:155 a common autoimmune disease 129:155 a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease 129:288 IgA nephropathy (IgAN) is a common autoimmune disease that is characterized by formation and deposition of IgA1-containing immune complexes frequently leading to end-stage kidney disease. 27333379 4 59 theme O-glycans 674:682 arg1 structure-function-studies 633:658 specific structure-function-studies 624:658 specific structure-function-studies of mucin-type O-glycans 624:682 Even though the disease is associated with the overproduction of aberrant O-glycans on IgA1, specific structure-function-studies of mucin-type O-glycans are limited. 26572920 2 0 theme N-glycan 508:515 arg1 release 517:523 N-glycan release 508:523 N-glycan release 508:523 FFPE samples were first deparaffinized, followed by solubilization in radioimmunoprecipitation assay buffer and treated with PNGase F for N-glycan release. 26572920 1 1 theme formaldehyde 329:340 arg1 treatment 342:350 formaldehyde treatment 329:350 formaldehyde treatment 329:350 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 2 theme treated 153:159 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 4 3 from any 737:739 arg1 profile 726:732 the N-glycome profile 712:732 the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 712:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 6 4 theme based 1047:1051 arg1 sequencing 1066:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 5 5 theme N-glycome 913:921 arg1 analysis 923:930 global N-glycome analysis 906:930 global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies 906:1029 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 3 6 theme released 530:537 arg1 glycans 539:545 The released glycans 526:545 The released glycans 526:545 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 1 7 theme mouse 245:249 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 7 theme mouse 245:249 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 6 8 theme N-linked 1148:1155 arg1 carbohydrates 1157:1169 the N-linked carbohydrates 1144:1169 the N-linked carbohydrates of the mouse tumor tissue samples 1144:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 1 9 gly glycoproteins 214:226 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 5 10 theme global 906:911 arg1 analysis 923:930 global N-glycome analysis 906:930 global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies 906:1029 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 1 11 theme tumor 251:255 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 11 theme tumor 251:255 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 3 12 with electrophoresis 613:627 arg1 detection 660:668 laser induced fluorescent detection 634:668 laser induced fluorescent detection 634:668 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 5 13 theme prospective 993:1003 arg1 studies 1023:1029 prospective and retrospective studies 993:1029 prospective and retrospective studies 993:1029 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 3 14 theme charged 567:573 arg1 fluorophore 575:585 a charged fluorophore 565:585 a charged fluorophore 565:585 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 1 15 theme tissue 257:262 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 15 theme tissue 257:262 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 16 dep susceptibility 310:323 arg1 mediated 352:359 mediated 352:359 mediated changes 352:367 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 4 17 theme tissue 812:817 arg1 samples 819:825 tissue samples 812:825 tissue samples 812:825 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 6 18 theme structural 1115:1124 arg1 information 1126:1136 some basic structural information 1104:1136 some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples 1104:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 4 19 gly glycoprotein 788:799 arg1 glycoprotein 788:799 glycoprotein 788:799 glycoprotein 788:799 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 5 20 theme retrospective 1009:1021 arg1 studies 1023:1029 prospective and retrospective studies 993:1029 prospective and retrospective studies 993:1029 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 1 21 theme fixed 174:178 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 4 22 dep glycoprotein 788:799 arg1 i.e. 782:785 i.e. 782:785 i.e. 782:785 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 5 23 used used 897:900 arg2 samples 874:880 FFPE samples 869:880 FFPE samples 869:880 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 4 24 theme N-glycome 716:724 arg1 profile 726:732 the N-glycome profile 712:732 the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 712:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 0 25 theme N-Glycosylation 0:14 arg1 analysis 16:23 N-Glycosylation analysis 0:23 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 0:97 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 26572920 4 26 theme complexation 761:772 arg1 levels 774:779 the investigated complexation levels 744:779 the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 744:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 5 27 theme novel 960:964 arg1 biomarkers 979:988 novel carbohydrate biomarkers 960:988 novel carbohydrate biomarkers 960:988 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 1 28 theme embedded 189:196 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 2 29 theme assay 465:469 arg1 buffer 471:476 radioimmunoprecipitation assay buffer 440:476 radioimmunoprecipitation assay buffer 440:476 FFPE samples were first deparaffinized, followed by solubilization in radioimmunoprecipitation assay buffer and treated with PNGase F for N-glycan release. 26572920 4 30 theme investigated 748:759 arg1 levels 774:779 the investigated complexation levels 744:779 the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 744:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 5 31 theme carbohydrate 966:977 arg1 biomarkers 979:988 novel carbohydrate biomarkers 960:988 novel carbohydrate biomarkers 960:988 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 4 32 theme significant 674:684 arg1 alterations 686:696 No significant alterations 671:696 No significant alterations 671:696 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 2 33 theme radioimmunoprecipitation 440:463 arg1 buffer 471:476 radioimmunoprecipitation assay buffer 440:476 radioimmunoprecipitation assay buffer 440:476 FFPE samples were first deparaffinized, followed by solubilization in radioimmunoprecipitation assay buffer and treated with PNGase F for N-glycan release. 26572920 4 34 located found 703:707 arg1 profile 726:732 the N-glycome profile 712:732 the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 712:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 4 34 located found 703:707 arg2 alterations 686:696 No significant alterations 671:696 No significant alterations 671:696 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 6 35 theme samples 1197:1203 arg1 carbohydrates 1157:1169 the N-linked carbohydrates 1144:1169 the N-linked carbohydrates of the mouse tumor tissue samples 1144:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 6 36 theme basic 1109:1113 arg1 information 1126:1136 some basic structural information 1104:1136 some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples 1104:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 5 37 theme FFPE 869:872 arg1 samples 874:880 FFPE samples 869:880 FFPE samples 869:880 These results suggest that FFPE samples can be readily used for global N-glycome analysis holding the promise to find novel carbohydrate biomarkers in prospective and retrospective studies. 26572920 0 38 theme fixed 37:41 arg1 samples 61:67 formalin fixed paraffin embedded samples 28:67 formalin fixed paraffin embedded samples 28:67 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 26572920 2 39 theme PNGase 495:500 arg1 F 502:502 PNGase F 495:502 PNGase F 495:502 FFPE samples were first deparaffinized, followed by solubilization in radioimmunoprecipitation assay buffer and treated with PNGase F for N-glycan release. 26572920 6 40 theme Exoglycosidase 1032:1045 arg1 sequencing 1066:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 1 41 dep embedded 189:196 arg1 FFPE 199:202 FFPE 199:202 FFPE 199:202 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 42 theme standard 205:212 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 0 43 theme samples 61:67 arg1 analysis 16:23 N-Glycosylation analysis 0:23 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 0:97 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 26572920 3 44 theme capillary 603:611 arg1 electrophoresis 613:627 capillary electrophoresis 603:627 capillary electrophoresis with laser induced fluorescent detection 603:668 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 1 45 from glycoproteins 214:226 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 45 from glycoproteins 214:226 arg1 serum 235:239 human serum 229:239 human serum 229:239 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 45 from glycoproteins 214:226 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 46 theme N-linked 114:121 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 46 theme N-linked 114:121 arg1 serum 235:239 human serum 229:239 human serum 229:239 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 46 theme N-linked 114:121 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 6 47 link N-linked 1148:1155 arg1 carbohydrates 1157:1169 the N-linked carbohydrates 1144:1169 the N-linked carbohydrates of the mouse tumor tissue samples 1144:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 0 48 dep fixed 37:41 arg1 embedded 52:59 embedded 52:59 embedded 52:59 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 26572920 6 49 theme tumor 1184:1188 arg1 samples 1197:1203 the mouse tumor tissue samples 1174:1203 the mouse tumor tissue samples 1174:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 1 50 link N-linked 114:121 arg1 samples 264:270 mouse tumor tissue samples 245:270 mouse tumor tissue samples 245:270 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 50 link N-linked 114:121 arg1 serum 235:239 human serum 229:239 human serum 229:239 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 50 link N-linked 114:121 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 4 51 theme study 835:839 arg1 levels 774:779 the investigated complexation levels 744:779 the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study 744:839 No significant alterations were found in the N-glycome profile at any of the investigated complexation levels (i.e., glycoprotein, serum and tissue samples) of the study. 26572920 6 52 theme mouse 1178:1182 arg1 samples 1197:1203 the mouse tumor tissue samples 1174:1203 the mouse tumor tissue samples 1174:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 1 53 theme intact 136:141 arg1 glycoproteins 214:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 136:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 6 54 theme tissue 1190:1195 arg1 samples 1197:1203 the mouse tumor tissue samples 1174:1203 the mouse tumor tissue samples 1174:1203 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 26572920 0 55 theme capillary 72:80 arg1 electrophoresis 82:96 capillary electrophoresis 72:96 capillary electrophoresis 72:96 N-Glycosylation analysis of formalin fixed paraffin embedded samples by capillary electrophoresis. 26572920 3 56 theme induced 640:646 arg1 detection 660:668 laser induced fluorescent detection 634:668 laser induced fluorescent detection 634:668 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 1 57 theme human 229:233 arg1 serum 235:239 human serum 229:239 human serum 229:239 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 1 57 theme human 229:233 arg1 glycans 123:129 N-linked glycans 114:129 N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins 114:226 In this study, N-linked glycans from intact, formalin treated and formalin fixed paraffin embedded (FFPE) standard glycoproteins, human serum and mouse tumor tissue samples were investigated in respect to their susceptibility for formaldehyde treatment mediated changes. 26572920 3 58 theme fluorescent 648:658 arg1 detection 660:668 laser induced fluorescent detection 634:668 laser induced fluorescent detection 634:668 The released glycans were labeled with a charged fluorophore and analyzed by capillary electrophoresis with laser induced fluorescent detection. 26572920 2 59 theme FFPE 370:373 arg1 samples 375:381 FFPE samples 370:381 FFPE samples 370:381 FFPE samples were first deparaffinized, followed by solubilization in radioimmunoprecipitation assay buffer and treated with PNGase F for N-glycan release. 26572920 6 60 theme carbohydrate 1053:1064 arg1 sequencing 1066:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing 1032:1075 Exoglycosidase based carbohydrate sequencing was also applied to reveal some basic structural information about the N-linked carbohydrates of the mouse tumor tissue samples. 25098670 0 0 theme thiol 57:61 arg1 synthesis 10:18 Expedient synthesis 0:18 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol 0:61 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol. 25098670 4 1 theme great 498:502 arg1 utility 504:510 the great utility 494:510 the great utility of MMTr as an effective anomeric S-protecting group 494:562 This work also demonstrated the great utility of MMTr as an effective anomeric S-protecting group. 25098670 3 2 theme linkages 332:339 arg1 α-configuration 302:316 The α-configuration 298:316 The α-configuration of glycosidic linkages 298:339 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 2 3 theme α-glycosyl 247:256 arg1 thiols 258:263 α-glycosyl thiols 247:263 α-glycosyl thiols 247:263 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 3 4 theme intrinsic 408:416 arg1 stability 434:442 the intrinsic configurational stability 404:442 the intrinsic configurational stability of α-glycosyl thiols 404:463 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 0 5 theme Expedient 0:8 arg1 synthesis 10:18 Expedient synthesis 0:18 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol 0:61 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol. 25098670 1 6 theme convenient 130:139 arg1 way 161:163 a convenient and stereoselective way 128:163 a convenient and stereoselective way 128:163 An α-S-(1→6)-linked pentaglucosyl thiol has been synthesized in a convenient and stereoselective way. 25098670 3 7 theme coupling 374:381 arg1 steps 383:387 the coupling steps 370:387 the coupling steps 370:387 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 4 8 theme anomeric 536:543 arg1 group 558:562 an effective anomeric S-protecting group 523:562 an effective anomeric S-protecting group 523:562 This work also demonstrated the great utility of MMTr as an effective anomeric S-protecting group. 25098670 1 9 theme stereoselective 145:159 arg1 way 161:163 a convenient and stereoselective way 128:163 a convenient and stereoselective way 128:163 An α-S-(1→6)-linked pentaglucosyl thiol has been synthesized in a convenient and stereoselective way. 25098670 2 10 gly thioglycosylation 202:218 arg1 sugars 235:240 6-iodinated sugars 223:240 6-iodinated sugars 223:240 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 4 11 theme effective 526:534 arg1 group 558:562 an effective anomeric S-protecting group 523:562 an effective anomeric S-protecting group 523:562 This work also demonstrated the great utility of MMTr as an effective anomeric S-protecting group. 25098670 2 12 theme transfer 277:284 arg1 conditions 286:295 phase transfer conditions 271:295 phase transfer conditions 271:295 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 3 13 theme α-glycosyl 447:456 arg1 thiols 458:463 α-glycosyl thiols 447:463 α-glycosyl thiols 447:463 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 2 14 theme phase 271:275 arg1 conditions 286:295 phase transfer conditions 271:295 phase transfer conditions 271:295 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 3 15 theme thiols 458:463 arg1 stability 434:442 the intrinsic configurational stability 404:442 the intrinsic configurational stability of α-glycosyl thiols 404:463 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 4 16 theme S-protecting 545:556 arg1 group 558:562 an effective anomeric S-protecting group 523:562 an effective anomeric S-protecting group 523:562 This work also demonstrated the great utility of MMTr as an effective anomeric S-protecting group. 25098670 4 17 theme MMTr 515:518 arg1 utility 504:510 the great utility 494:510 the great utility of MMTr as an effective anomeric S-protecting group 494:562 This work also demonstrated the great utility of MMTr as an effective anomeric S-protecting group. 25098670 2 18 theme synthesis 183:191 arg1 steps 170:174 Key steps 166:174 Key steps of the synthesis 166:191 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 2 19 with thioglycosylation 202:218 arg1 thiols 258:263 α-glycosyl thiols 247:263 α-glycosyl thiols 247:263 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 3 20 theme glycosidic 321:330 arg1 linkages 332:339 glycosidic linkages 321:339 glycosidic linkages 321:339 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 1 21 theme -linked 76:82 arg1 thiol 98:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol has been synthesized in a convenient and stereoselective way. 25098670 2 22 theme Key 166:168 arg1 steps 170:174 Key steps 166:174 Key steps of the synthesis 166:191 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 2 23 theme sugars 235:240 arg1 thioglycosylation 202:218 thioglycosylation 202:218 thioglycosylation of 6-iodinated sugars with α-glycosyl thiols 202:263 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 0 24 link -linked 35:41 arg1 thiol 57:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol. 25098670 1 25 theme pentaglucosyl 84:96 arg1 thiol 98:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol has been synthesized in a convenient and stereoselective way. 25098670 1 26 link -linked 76:82 arg1 thiol 98:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol 64:102 An α-S-(1→6)-linked pentaglucosyl thiol has been synthesized in a convenient and stereoselective way. 25098670 2 27 theme 6-iodinated 223:233 arg1 sugars 235:240 6-iodinated sugars 223:240 6-iodinated sugars 223:240 Key steps of the synthesis involved thioglycosylation of 6-iodinated sugars with α-glycosyl thiols under phase transfer conditions. 25098670 0 28 theme pentaglucosyl 43:55 arg1 thiol 57:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol. 25098670 3 29 theme configurational 418:432 arg1 stability 434:442 the intrinsic configurational stability 404:442 the intrinsic configurational stability of α-glycosyl thiols 404:463 The α-configuration of glycosidic linkages was thus introduced prior to the coupling steps, and relied on the intrinsic configurational stability of α-glycosyl thiols. 25098670 0 30 theme -linked 35:41 arg1 thiol 57:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 an α-S-(1→6)-linked pentaglucosyl thiol 23:61 Expedient synthesis of an α-S-(1→6)-linked pentaglucosyl thiol. 25310684 4 0 theme β-d-galactofuranosyl 766:785 arg1 tri- 787:790 β-d-galactofuranosyl tri- 766:790 β-d-galactofuranosyl tri- 766:790 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 0 1 theme thioglycosides 78:91 arg1 application 97:107 application 97:107 application of the silylated 97:124 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 0 1 theme thioglycosides 78:91 arg1 reactivity 47:56 the anomeric reactivity 34:56 the anomeric reactivity of galactofuranosyl thioglycosides 34:91 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 0 2 from Influence 0:8 arg1 application 97:107 application 97:107 application of the silylated 97:124 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 0 2 from Influence 0:8 arg1 reactivity 47:56 the anomeric reactivity 34:56 the anomeric reactivity of galactofuranosyl thioglycosides 34:91 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 2 3 from ethers 354:359 arg1 ring 384:387 the galactofuranose ring 364:387 the galactofuranose ring 364:387 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 1 4 theme donor 288:292 arg1 reactivity 294:303 the donor reactivity 284:303 the donor reactivity of galactofuranosyl phenyl thioglycosides 284:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 3 5 theme useful 620:625 arg1 block 645:649 a useful glycosyl building block 618:649 a useful glycosyl building block 618:649 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 3 5 theme useful 620:625 arg1 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 546:598 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 0 6 theme silylated 116:124 arg1 application 97:107 application 97:107 application of the silylated 97:124 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 0 6 theme silylated 116:124 arg1 reactivity 47:56 the anomeric reactivity 34:56 the anomeric reactivity of galactofuranosyl thioglycosides 34:91 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 4 7 contain possessing 813:822 arg1 tetrasaccharides 796:811 tetrasaccharides 796:811 tetrasaccharides possessing diverse glycosidic linkages 796:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 4 7 contain possessing 813:822 arg2 linkages 843:850 diverse glycosidic linkages 824:850 diverse glycosidic linkages 824:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 4 8 theme solution-phase 728:741 arg1 assembly 743:750 the highly efficient one-pot solution-phase assembly 699:750 the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 699:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 2 9 contain have 402:405 arg1 ethers 354:359 Silyl ethers 348:359 Silyl ethers on the galactofuranose ring 348:387 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 2 9 contain have 402:405 arg2 effect 417:422 an arming effect 407:422 an arming effect on the glycosylation reactivity 407:454 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 4 10 theme one-pot 720:726 arg1 assembly 743:750 the highly efficient one-pot solution-phase assembly 699:750 the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 699:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 2 11 theme protecting 483:492 arg1 group 494:498 the cyclic 3,5-acetal protecting group 461:498 the cyclic 3,5-acetal protecting group 461:498 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 4 12 theme glycosidic 832:841 arg1 linkages 843:850 diverse glycosidic linkages 824:850 diverse glycosidic linkages 824:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 2 13 theme 3,5-acetal 472:481 arg1 group 494:498 the cyclic 3,5-acetal protecting group 461:498 the cyclic 3,5-acetal protecting group 461:498 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 4 14 theme diverse 824:830 arg1 linkages 843:850 diverse glycosidic linkages 824:850 diverse glycosidic linkages 824:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 2 15 theme arming 410:415 arg1 effect 417:422 an arming effect 407:422 an arming effect on the glycosylation reactivity 407:454 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 0 16 theme protections 19:29 arg1 Influence 0:8 Influence 0:8 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated 0:124 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 1 17 theme galactofuranosyl 308:323 arg1 thioglycosides 332:345 galactofuranosyl phenyl thioglycosides 308:345 galactofuranosyl phenyl thioglycosides 308:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 2 18 theme Silyl 348:352 arg1 ethers 354:359 Silyl ethers 348:359 Silyl ethers on the galactofuranose ring 348:387 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 0 19 theme silyl 13:17 arg1 protections 19:29 silyl protections 13:29 silyl protections 13:29 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 1 20 theme phenyl 325:330 arg1 thioglycosides 332:345 galactofuranosyl phenyl thioglycosides 308:345 galactofuranosyl phenyl thioglycosides 308:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 3 21 theme building 636:643 arg1 block 645:649 a useful glycosyl building block 618:649 a useful glycosyl building block 618:649 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 3 21 theme building 636:643 arg1 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 546:598 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 1 22 theme thioglycosides 332:345 arg1 reactivity 294:303 the donor reactivity 284:303 the donor reactivity of galactofuranosyl phenyl thioglycosides 284:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 4 23 theme efficient 710:718 arg1 assembly 743:750 the highly efficient one-pot solution-phase assembly 699:750 the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 699:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 1 24 theme tuning 240:245 arg1 effect 247:252 the tuning effect 236:252 the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides 236:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 0 25 theme one-pot 152:158 arg1 synthesis 160:168 one-pot synthesis 152:168 one-pot synthesis of diverse β-D-oligogalactofuranosides 152:207 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 4 26 theme tri- 787:790 arg1 panel 757:761 a panel 755:761 a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 755:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 2 27 theme cyclic 465:470 arg1 group 494:498 the cyclic 3,5-acetal protecting group 461:498 the cyclic 3,5-acetal protecting group 461:498 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 2 28 from effect 417:422 arg1 reactivity 445:454 the glycosylation reactivity 427:454 the glycosylation reactivity 427:454 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 2 29 theme glycosylation 431:443 arg1 reactivity 445:454 the glycosylation reactivity 427:454 the glycosylation reactivity 427:454 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 0 30 theme diverse 173:179 arg1 β-D-oligogalactofuranosides 181:207 diverse β-D-oligogalactofuranosides 173:207 diverse β-D-oligogalactofuranosides 173:207 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 1 31 from effect 247:252 arg1 reactivity 294:303 the donor reactivity 284:303 the donor reactivity of galactofuranosyl phenyl thioglycosides 284:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 3 32 theme reactive 530:537 arg1 block 645:649 a useful glycosyl building block 618:649 a useful glycosyl building block 618:649 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 3 32 theme reactive 530:537 arg1 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 546:598 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 0 33 theme anomeric 38:45 arg1 reactivity 47:56 the anomeric reactivity 34:56 the anomeric reactivity of galactofuranosyl thioglycosides 34:91 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 3 34 theme phenyl 539:544 arg1 block 645:649 a useful glycosyl building block 618:649 a useful glycosyl building block 618:649 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 3 34 theme phenyl 539:544 arg1 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 546:598 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 2 35 theme galactofuranose 368:382 arg1 ring 384:387 the galactofuranose ring 364:387 the galactofuranose ring 364:387 Silyl ethers on the galactofuranose ring are found to have an arming effect on the glycosylation reactivity, but the cyclic 3,5-acetal protecting group decreases the reactivity. 25310684 3 36 theme glycosyl 627:634 arg1 block 645:649 a useful glycosyl building block 618:649 a useful glycosyl building block 618:649 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 3 36 theme glycosyl 627:634 arg1 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 546:598 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 526:600 The reactive phenyl 2,6-di-O-Bz-3,5-di-O-TBS-1-thio-β-d-galactofuranoside 3 is proved to be a useful glycosyl building block. 25310684 4 37 theme donor 680:684 arg1 advantage 662:670 advantage 662:670 advantage of this donor 662:684 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 0 38 theme β-D-oligogalactofuranosides 181:207 arg1 synthesis 160:168 one-pot synthesis 152:168 one-pot synthesis of diverse β-D-oligogalactofuranosides 152:207 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 4 39 theme tetrasaccharides 796:811 arg1 panel 757:761 a panel 755:761 a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 755:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 1 40 theme silyl 257:261 arg1 groups 274:279 silyl protecting groups 257:279 silyl protecting groups 257:279 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 0 41 theme galactofuranosyl 61:76 arg1 thioglycosides 78:91 galactofuranosyl thioglycosides 61:91 galactofuranosyl thioglycosides 61:91 Influence of silyl protections on the anomeric reactivity of galactofuranosyl thioglycosides and application of the silylated thiogalactofuranosides to one-pot synthesis of diverse β-D-oligogalactofuranosides. 25310684 4 42 theme panel 757:761 arg1 assembly 743:750 the highly efficient one-pot solution-phase assembly 699:750 the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages 699:850 By taking advantage of this donor, we achieved the highly efficient one-pot solution-phase assembly of a panel of β-d-galactofuranosyl tri- and tetrasaccharides possessing diverse glycosidic linkages. 25310684 1 43 theme protecting 263:272 arg1 groups 274:279 silyl protecting groups 257:279 silyl protecting groups 257:279 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 25310684 1 44 theme groups 274:279 arg1 effect 247:252 the tuning effect 236:252 the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides 236:345 We describe in this paper the tuning effect of silyl protecting groups on the donor reactivity of galactofuranosyl phenyl thioglycosides. 24945257 5 0 theme protein 822:828 arg1 structures 830:839 identical protein structures 812:839 identical protein structures 812:839 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 1 1 attach isolated 128:135 arg2 Acutobin 119:126 Acutobin 119:126 Acutobin isolated from Deinagkistrodon acutus venom 119:169 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 1 1 attach isolated 128:135 arg1 venom 165:169 Deinagkistrodon acutus venom 142:169 Deinagkistrodon acutus venom 142:169 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 10 2 theme clinical 1812:1819 arg1 use 1821:1823 clinical use 1812:1823 clinical use 1812:1823 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 9 3 dep in 1414:1415 arg1 vivo 1417:1420 vivo 1417:1420 vivo 1417:1420 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 4 4 from level 535:539 arg1 mice 579:582 mice 579:582 mice after i.p.-injection of desialylated-acutobin 579:628 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 7 5 theme Bβ 1159:1160 arg1 chains 1168:1173 the Bβ and γ chains 1155:1173 chains 1168:1173 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 10 6 with interaction 1594:1604 arg1 fibrinogen 1611:1620 fibrinogen 1611:1620 fibrinogen 1611:1620 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 4 7 theme desialylated-acutobin 608:628 arg1 i.p.-injection 590:603 i.p.-injection 590:603 i.p.-injection of desialylated-acutobin 590:628 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 3 8 theme catalytic 389:397 arg1 activities 399:408 similar catalytic activities 381:408 similar catalytic activities 381:408 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 2 9 theme 39 269:270 arg1 kDa 272:274 kDa 272:274 kDa 272:274 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 10 theme disialyl-capped 309:323 arg1 N-glycans 325:333 terminal disialyl-capped N-glycans 300:333 terminal disialyl-capped N-glycans 300:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 0 11 theme recombinant 95:105 arg1 glycoforms 107:116 its recombinant glycoforms 91:116 its recombinant glycoforms 91:116 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 5 12 with activities 864:873 arg1 structures 830:839 identical protein structures 812:839 identical protein structures 812:839 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 2 13 theme terminal 300:307 arg1 N-glycans 325:333 terminal disialyl-capped N-glycans 300:333 terminal disialyl-capped N-glycans 300:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 9 14 theme HKATB 1450:1454 arg1 effect 1440:1445 The in vivo defibrinogenating effect 1410:1445 The in vivo defibrinogenating effect of HKATB 1410:1454 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 9 14 theme HKATB 1450:1454 arg1 inferior 1460:1467 inferior 1460:1467 inferior 1460:1467 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 5 15 theme HEK293T 923:929 arg1 cells 942:946 HEK293T and SW1353 cells 923:946 cells 942:946 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 2 16 gly glycoprotein 276:287 arg1 glycoprotein 276:287 a 39 kDa glycoprotein 267:287 a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans 267:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 16 gly glycoprotein 276:287 arg1 protease 255:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 10 17 theme conjugated 1543:1552 arg1 glycans 1554:1560 the conjugated glycans 1539:1560 the conjugated glycans of acutobin 1539:1572 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 4 18 theme acutobin 675:682 arg1 injection 684:692 acutobin injection 675:692 acutobin injection 675:692 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 8 19 theme peptide 1305:1311 arg1 F 1327:1327 peptide N-glycosidase F 1305:1327 peptide N-glycosidase F 1305:1327 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 3 20 theme fibrinogen 468:477 arg1 chain 459:463 the Aα chain 452:463 the Aα chain of fibrinogen as efficiently as the native acutobin did 452:519 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 3 21 theme similar 381:387 arg1 activities 399:408 similar catalytic activities 381:408 similar catalytic activities 381:408 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 7 22 theme human 1178:1182 arg1 fibrinogens 1184:1194 human fibrinogens 1178:1194 human fibrinogens 1178:1194 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 5 23 theme recombinant 787:797 arg1 HKATB 966:970 HKATB 966:970 HKATB 966:970 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 5 23 theme recombinant 787:797 arg1 enzymes 799:805 Two recombinant enzymes 783:805 Two recombinant enzymes with identical protein structures 783:839 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 5 23 theme recombinant 787:797 arg1 SWATB 976:980 SWATB 976:980 SWATB 976:980 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 10 24 theme efficient 1676:1684 arg1 glycoforms 1686:1695 efficient glycoforms 1676:1695 efficient glycoforms 1676:1695 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 7 25 theme Aα 1137:1138 arg1 chain 1140:1144 the Aα chain 1133:1144 not only the Aα chain but also the Bβ and γ chains of human fibrinogens 1124:1194 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 9 26 theme in 1414:1415 arg1 effect 1440:1445 The in vivo defibrinogenating effect 1410:1445 The in vivo defibrinogenating effect of HKATB 1410:1454 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 9 26 theme in 1414:1415 arg1 inferior 1460:1467 inferior 1460:1467 inferior 1460:1467 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 9 27 from acutobin 1480:1487 arg1 mice 1492:1495 mice 1492:1495 mice 1492:1495 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 5 28 theme SW1353 935:940 arg1 cells 942:946 HEK293T and SW1353 cells 923:946 cells 942:946 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 7 29 theme fibrinogens 1184:1194 arg1 chains 1168:1173 the Bβ and γ chains 1155:1173 chains 1168:1173 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 7 29 theme fibrinogens 1184:1194 arg1 chain 1140:1144 the Aα chain 1133:1144 not only the Aα chain but also the Bβ and γ chains of human fibrinogens 1124:1194 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 8 30 theme proteases 1292:1300 arg1 deglycosylation 1267:1281 Non-denaturing deglycosylation 1252:1281 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F 1252:1327 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 8 31 gly deglycosylation 1267:1281 arg1 proteases 1292:1300 these proteases 1286:1300 these proteases 1286:1300 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 0 32 gly glycoforms 107:116 arg1 acutobin 78:85 acutobin 78:85 acutobin 78:85 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 8 33 theme N-glycosidase 1313:1325 arg1 F 1327:1327 peptide N-glycosidase F 1305:1327 peptide N-glycosidase F 1305:1327 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 4 34 theme disialyl 715:722 arg1 moieties 724:731 the disialyl moieties 711:731 the disialyl moieties 711:731 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 0 35 theme glycan 24:29 arg1 variations 31:40 glycan variations 24:40 glycan variations 24:40 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 2 36 contain containing 289:298 arg1 glycoprotein 276:287 a 39 kDa glycoprotein 267:287 a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans 267:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 36 contain containing 289:298 arg1 protease 255:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 36 contain containing 289:298 arg2 N-glycans 325:333 terminal disialyl-capped N-glycans 300:333 terminal disialyl-capped N-glycans 300:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 8 37 theme Non-denaturing 1252:1265 arg1 deglycosylation 1267:1281 Non-denaturing deglycosylation 1252:1281 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F 1252:1327 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 3 38 theme chromogenic 417:427 arg1 substrate 429:437 chromogenic substrate 417:437 chromogenic substrate 417:437 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 9 39 theme defibrinogenating 1422:1438 arg1 effect 1440:1445 The in vivo defibrinogenating effect 1410:1445 The in vivo defibrinogenating effect of HKATB 1410:1454 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 9 39 theme defibrinogenating 1422:1438 arg1 inferior 1460:1467 inferior 1460:1467 inferior 1460:1467 The in vivo defibrinogenating effect of HKATB was inferior to that of acutobin in mice. 24945257 10 40 theme acutobin 1565:1572 arg1 glycans 1554:1560 the conjugated glycans 1539:1560 the conjugated glycans of acutobin 1539:1572 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 3 41 theme native 501:506 arg1 acutobin 508:515 the native acutobin 497:515 the Aα chain of fibrinogen as efficiently as the native acutobin did 452:519 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 7 42 theme α-fibrinogenase 1226:1240 arg1 activity 1242:1249 a reduced α-fibrinogenase activity 1216:1249 a reduced α-fibrinogenase activity 1216:1249 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 10 43 theme native 1794:1799 arg1 enzyme 1801:1806 the native enzyme 1790:1806 the native enzyme for clinical use 1790:1823 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 10 44 theme cells 1649:1653 arg1 desirable 1739:1747 desirable 1739:1747 desirable 1739:1747 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 10 44 theme cells 1649:1653 arg1 selection 1636:1644 the selection 1632:1644 the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering 1632:1733 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 10 45 theme recombinant 1758:1768 arg1 product 1770:1776 a recombinant product 1756:1776 a recombinant product 1756:1776 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 3 46 theme Aα 456:457 arg1 chain 459:463 the Aα chain 452:463 the Aα chain of fibrinogen as efficiently as the native acutobin did 452:519 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 4 47 theme degradation 555:565 arg1 products 567:574 fibrinogen degradation products 544:574 fibrinogen degradation products 544:574 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 5 48 theme similar 845:851 arg1 activities 864:873 similar amidolytic activities 845:873 similar amidolytic activities to those of native acutobin 845:901 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 2 49 theme serine 248:253 arg1 glycoprotein 276:287 a 39 kDa glycoprotein 267:287 a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans 267:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 49 theme serine 248:253 arg1 protease 255:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 7 50 theme reduced 1218:1224 arg1 activity 1242:1249 a reduced α-fibrinogenase activity 1216:1249 a reduced α-fibrinogenase activity 1216:1249 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 5 51 theme amidolytic 853:862 arg1 activities 864:873 similar amidolytic activities 845:873 similar amidolytic activities to those of native acutobin 845:901 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 4 52 theme fibrinogen 544:553 arg1 products 567:574 fibrinogen degradation products 544:574 fibrinogen degradation products 544:574 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 2 53 theme defibrinogenating 230:246 arg1 glycoprotein 276:287 a 39 kDa glycoprotein 267:287 a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans 267:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 53 theme defibrinogenating 230:246 arg1 protease 255:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 8 54 theme fibrinogenolytic 1357:1372 arg1 activities 1374:1383 their fibrinogenolytic activities 1351:1383 their fibrinogenolytic activities 1351:1383 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 0 55 theme defibrinogenating 46:62 arg1 activities 64:73 defibrinogenating activities 46:73 defibrinogenating activities 46:73 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 6 56 theme Mass 997:1000 arg1 profiling 1016:1024 Mass spectrometric profiling 997:1024 Mass spectrometric profiling 997:1024 Mass spectrometric profiling showed that their glycans differed from those of acutobin. 24945257 3 57 theme sialidase 342:350 arg1 treatment 352:360 sialidase treatment 342:360 sialidase treatment 342:360 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 0 58 dep variations 31:40 arg1 the 20:22 the 20:22 the 20:22 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 1 59 from stroke 205:210 arg1 patients 215:222 patients 215:222 patients 215:222 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 7 60 theme γ 1166:1166 arg1 chains 1168:1173 the Bβ and γ chains 1155:1173 chains 1168:1173 In contrast to acutobin, HKATB cleaved not only the Aα chain but also the Bβ and γ chains of human fibrinogens, while SWATB showed a reduced α-fibrinogenase activity. 24945257 4 61 theme products 567:574 arg1 lower 648:652 lower 648:652 lower 648:652 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 4 61 theme products 567:574 arg1 level 535:539 the level 531:539 the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin 531:628 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 10 62 theme glycosylation 1709:1721 arg1 engineering 1723:1733 further glycosylation engineering 1701:1733 further glycosylation engineering 1701:1733 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 3 63 dep efficiently 482:492 arg1 as 494:495 as 494:495 as 494:495 After sialidase treatment, the enzyme showed similar catalytic activities toward chromogenic substrate, and cleaved the Aα chain of fibrinogen as efficiently as the native acutobin did. 24945257 2 64 theme kDa 272:274 arg1 glycoprotein 276:287 a 39 kDa glycoprotein 267:287 a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans 267:333 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 2 64 theme kDa 272:274 arg1 protease 255:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease 225:262 This defibrinogenating serine protease is a 39 kDa glycoprotein containing terminal disialyl-capped N-glycans. 24945257 5 65 with enzymes 799:805 arg1 structures 830:839 identical protein structures 812:839 identical protein structures 812:839 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 1 66 theme Deinagkistrodon 142:156 arg1 venom 165:169 Deinagkistrodon acutus venom 142:169 Deinagkistrodon acutus venom 142:169 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 1 67 used used 180:183 arg2 Acutobin 119:126 Acutobin 119:126 Acutobin isolated from Deinagkistrodon acutus venom 119:169 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 1 68 theme acutus 158:163 arg1 venom 165:169 Deinagkistrodon acutus venom 142:169 Deinagkistrodon acutus venom 142:169 Acutobin isolated from Deinagkistrodon acutus venom has been used to prevent or treat stroke in patients. 24945257 4 69 theme acutobin 773:780 arg1 half-life 760:768 the half-life 756:768 the half-life of acutobin 756:780 However, the level of fibrinogen degradation products in mice after i.p.-injection of desialylated-acutobin was significantly lower than the level after acutobin injection, suggesting that the disialyl moieties may improve or prolong the half-life of acutobin. 24945257 0 70 theme acutobin 78:85 arg1 glycoforms 107:116 its recombinant glycoforms 91:116 its recombinant glycoforms 91:116 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 0 70 theme acutobin 78:85 arg1 variations 31:40 glycan variations 24:40 glycan variations 24:40 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 0 70 theme acutobin 78:85 arg1 activities 64:73 defibrinogenating activities 46:73 defibrinogenating activities 46:73 Correlation between the glycan variations and defibrinogenating activities of acutobin and its recombinant glycoforms. 24945257 5 71 theme identical 812:820 arg1 structures 830:839 identical protein structures 812:839 identical protein structures 812:839 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 5 72 theme native 887:892 arg1 acutobin 894:901 native acutobin 887:901 native acutobin 887:901 Two recombinant enzymes with identical protein structures and similar amidolytic activities to those of native acutobin were expressed from HEK293T and SW1353 cells and designated as HKATB and SWATB, respectively. 24945257 8 73 theme thermal 1389:1395 arg1 stabilities 1397:1407 thermal stabilities 1389:1407 thermal stabilities 1389:1407 Non-denaturing deglycosylation of these proteases by peptide N-glycosidase F significantly reduced their fibrinogenolytic activities and thermal stabilities. 24945257 10 74 theme further 1701:1707 arg1 engineering 1723:1733 further glycosylation engineering 1701:1733 further glycosylation engineering 1701:1733 Taken together, our results suggest that the conjugated glycans of acutobin are involved in its interaction with fibrinogen, and that the selection of cells optimally expressing efficient glycoforms and further glycosylation engineering are desirable before a recombinant product can replace the native enzyme for clinical use. 24945257 6 75 theme spectrometric 1002:1014 arg1 profiling 1016:1024 Mass spectrometric profiling 997:1024 Mass spectrometric profiling 997:1024 Mass spectrometric profiling showed that their glycans differed from those of acutobin. 28089449 2 0 theme catalytic 509:517 arg1 apparatuses 519:529 highly similar catalytic apparatuses 494:529 highly similar catalytic apparatuses 494:529 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 5 1 theme catalytic 1212:1220 arg1 properties 1222:1231 desired catalytic properties 1204:1231 desired catalytic properties 1204:1231 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 2 2 theme cyclic 637:642 arg1 cycloalternan 660:672 the cyclic tetrasaccharide cycloalternan 633:672 the cyclic tetrasaccharide cycloalternan (CA) 633:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 2 theme cyclic 637:642 arg1 CA 675:676 CA 675:676 CA 675:676 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 3 3 theme stable 759:764 arg1 site 773:776 a conformationally stable active site 740:776 a conformationally stable active site 740:776 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 1 4 theme electrophile 234:245 arg1 transfer 219:226 the transfer 215:226 the transfer of an electrophile to a non-water nucleophile (transferases) 215:287 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 0 5 theme Reaction 72:79 arg1 Specificity 81:91 Reaction Specificity 72:91 Reaction Specificity 72:91 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 28089449 1 6 theme related 344:350 arg1 reactions 363:371 mechanistically related hydrolysis reactions 328:371 mechanistically related hydrolysis reactions 328:371 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 5 7 theme principles 1128:1137 arg1 Application 1107:1117 Application 1107:1117 Application of these principles 1107:1137 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 1 8 theme hydrolysis 352:361 arg1 reactions 363:371 mechanistically related hydrolysis reactions 328:371 mechanistically related hydrolysis reactions 328:371 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 4 9 theme classic 931:937 arg1 view 939:942 the classic view 927:942 the classic view of induced-fit conformational change 927:979 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 2 10 gly transglycosylation 547:564 arg1 linkages 621:628 α-1,3-glucan linkages 608:628 α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA) 608:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 3 11 theme change 866:871 arg1 pattern 828:834 a distinct pattern 817:834 a distinct pattern of active site conformational change 817:871 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 3 12 theme active 766:771 arg1 site 773:776 a conformationally stable active site 740:776 a conformationally stable active site 740:776 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 2 13 theme linkages 621:628 arg1 reaction 581:588 a transferase reaction 567:588 a transferase reaction 567:588 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 13 theme linkages 621:628 arg1 hydrolysis 594:603 hydrolysis 594:603 hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA) 594:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 13 theme linkages 621:628 arg1 transglycosylation 547:564 the transglycosylation 543:564 the transglycosylation (a transferase reaction) 543:589 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 1 14 theme non-water 252:260 arg1 transferases 275:286 transferases 275:286 transferases 275:286 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 1 14 theme non-water 252:260 arg1 nucleophile 262:272 a non-water nucleophile 250:272 a non-water nucleophile (transferases) 250:287 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 3 15 theme distinct 819:826 arg1 pattern 828:834 a distinct pattern 817:834 a distinct pattern of active site conformational change 817:871 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 5 16 with reengineering 1169:1181 arg1 properties 1222:1231 desired catalytic properties 1204:1231 desired catalytic properties 1204:1231 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 1 17 theme water 163:167 arg1 present 184:190 present 184:190 present 184:190 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 1 17 theme water 163:167 arg1 excess 153:158 a massive excess 143:158 a massive excess of water 143:167 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 0 18 from Role 34:37 arg1 Specificity 81:91 Reaction Specificity 72:91 Reaction Specificity 72:91 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 28089449 2 19 from hydrolysis 594:603 arg1 cycloalternan 660:672 the cyclic tetrasaccharide cycloalternan 633:672 the cyclic tetrasaccharide cycloalternan (CA) 633:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 19 from hydrolysis 594:603 arg1 CA 675:676 CA 675:676 CA 675:676 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 1 20 from environment 125:135 arg1 Active 94:99 Active 94:99 Active 94:99 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 4 21 theme disfavor 1086:1093 arg1 hydrolysis 1095:1104 disfavor hydrolysis 1086:1104 disfavor hydrolysis 1086:1104 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 2 22 theme similar 501:507 arg1 apparatuses 519:529 highly similar catalytic apparatuses 494:529 highly similar catalytic apparatuses 494:529 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 3 23 theme non-catalytic 714:726 arg1 domains 728:734 non-catalytic domains 714:734 non-catalytic domains 714:734 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 2 24 theme tetrasaccharide 644:658 arg1 cycloalternan 660:672 the cyclic tetrasaccharide cycloalternan 633:672 the cyclic tetrasaccharide cycloalternan (CA) 633:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 24 theme tetrasaccharide 644:658 arg1 CA 675:676 CA 675:676 CA 675:676 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 25 from linkages 621:628 arg1 cycloalternan 660:672 the cyclic tetrasaccharide cycloalternan 633:672 the cyclic tetrasaccharide cycloalternan (CA) 633:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 25 from linkages 621:628 arg1 CA 675:676 CA 675:676 CA 675:676 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 5 26 theme rational 1160:1167 arg1 reengineering 1169:1181 the rational reengineering 1156:1181 the rational reengineering of transferases with desired catalytic properties 1156:1231 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 4 27 theme change 974:979 arg1 view 939:942 the classic view 927:942 the classic view of induced-fit conformational change 927:979 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 4 28 theme binding 1038:1044 arg1 site 1046:1049 a stable hydrophobic binding site 1017:1049 a stable hydrophobic binding site 1017:1049 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 2 29 theme reaction 438:445 arg1 specificity 447:457 transferase versus hydrolase reaction specificity 409:457 transferase versus hydrolase reaction specificity 409:457 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 3 30 theme substrate 693:701 arg1 binding 703:709 substrate binding 693:709 substrate binding to non-catalytic domains 693:734 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 2 31 theme transferase 569:579 arg1 reaction 581:588 a transferase reaction 567:588 a transferase reaction 567:588 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 31 theme transferase 569:579 arg1 transglycosylation 547:564 the transglycosylation 543:564 the transglycosylation (a transferase reaction) 543:589 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 32 theme hydrolase 428:436 arg1 specificity 447:457 transferase versus hydrolase reaction specificity 409:457 transferase versus hydrolase reaction specificity 409:457 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 33 theme transferase 409:419 arg1 specificity 447:457 transferase versus hydrolase reaction specificity 409:457 transferase versus hydrolase reaction specificity 409:457 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 34 from cycloalternan 660:672 arg1 reaction 581:588 a transferase reaction 567:588 a transferase reaction 567:588 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 34 from cycloalternan 660:672 arg1 hydrolysis 594:603 hydrolysis 594:603 hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA) 594:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 34 from cycloalternan 660:672 arg1 transglycosylation 547:564 the transglycosylation 543:564 the transglycosylation (a transferase reaction) 543:589 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 35 theme a 567:567 arg1 reaction 581:588 a transferase reaction 567:588 a transferase reaction 567:588 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 35 theme a 567:567 arg1 transglycosylation 547:564 the transglycosylation 543:564 the transglycosylation (a transferase reaction) 543:589 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 0 36 theme Conformational 42:55 arg1 Flexibility 57:67 Conformational Flexibility 42:67 Conformational Flexibility 42:67 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 28089449 1 37 theme specific 297:304 arg1 strategies 306:315 specific strategies 297:315 specific strategies 297:315 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 2 38 theme α-1,3-glucan 608:619 arg1 linkages 621:628 α-1,3-glucan linkages 608:628 α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA) 608:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 4 39 theme stable 1019:1024 arg1 site 1046:1049 a stable hydrophobic binding site 1017:1049 a stable hydrophobic binding site 1017:1049 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 3 40 theme CA 786:787 arg1 transglycosylation 789:806 CA transglycosylation 786:806 CA transglycosylation 786:806 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 5 41 theme transferases 1186:1197 arg1 reengineering 1169:1181 the rational reengineering 1156:1181 the rational reengineering of transferases with desired catalytic properties 1156:1231 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 1 42 theme aqueous 108:114 arg1 environment 125:135 the aqueous cellular environment 104:135 the aqueous cellular environment where a massive excess of water is perpetually present 104:190 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 4 43 theme transferase 1061:1071 arg1 activity 1073:1080 transferase activity 1061:1080 transferase activity 1061:1080 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 5 44 theme desired 1204:1210 arg1 properties 1222:1231 desired catalytic properties 1204:1231 desired catalytic properties 1204:1231 Application of these principles could facilitate the rational reengineering of transferases with desired catalytic properties. 28089449 1 45 theme cellular 116:123 arg1 environment 125:135 the aqueous cellular environment 104:135 the aqueous cellular environment where a massive excess of water is perpetually present 104:190 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 4 46 theme conformational 959:972 arg1 change 974:979 induced-fit conformational change 947:979 induced-fit conformational change 947:979 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 3 47 theme site 846:849 arg1 change 866:871 active site conformational change 839:871 active site conformational change 839:871 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 4 48 theme induced-fit 947:957 arg1 change 974:979 induced-fit conformational change 947:979 induced-fit conformational change 947:979 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 3 49 theme CA 892:893 arg1 hydrolysis 895:904 CA hydrolysis 892:904 CA hydrolysis 892:904 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 3 50 theme conformational 851:864 arg1 change 866:871 active site conformational change 839:871 active site conformational change 839:871 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 4 51 theme hydrophobic 1026:1036 arg1 site 1046:1049 a stable hydrophobic binding site 1017:1049 a stable hydrophobic binding site 1017:1049 These findings defy the classic view of induced-fit conformational change and illustrate a mechanism by which a stable hydrophobic binding site can favor transferase activity and disfavor hydrolysis. 28089449 2 52 from transglycosylation 547:564 arg1 cycloalternan 660:672 the cyclic tetrasaccharide cycloalternan 633:672 the cyclic tetrasaccharide cycloalternan (CA) 633:677 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 2 52 from transglycosylation 547:564 arg1 CA 675:676 CA 675:676 CA 675:676 To identify principles that confer transferase versus hydrolase reaction specificity, we exploited two enzymes that use highly similar catalytic apparatuses to catalyze the transglycosylation (a transferase reaction) or hydrolysis of α-1,3-glucan linkages in the cyclic tetrasaccharide cycloalternan (CA). 28089449 1 53 from Active 94:99 arg1 environment 125:135 the aqueous cellular environment 104:135 the aqueous cellular environment where a massive excess of water is perpetually present 104:190 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 0 54 theme Flexibility 57:67 arg1 Role 34:37 The Role 30:37 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 0:92 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 28089449 3 55 dep promote 778:784 arg1 whereas 809:815 whereas 809:815 whereas 809:815 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28089449 0 56 dep Transferase 0:10 arg1 Role 34:37 The Role 30:37 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 0:92 Transferase Versus Hydrolase: The Role of Conformational Flexibility in Reaction Specificity. 28089449 1 57 theme massive 145:151 arg1 present 184:190 present 184:190 present 184:190 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 1 57 theme massive 145:151 arg1 excess 153:158 a massive excess 143:158 a massive excess of water 143:167 Active in the aqueous cellular environment where a massive excess of water is perpetually present, enzymes that catalyze the transfer of an electrophile to a non-water nucleophile (transferases) require specific strategies to inhibit mechanistically related hydrolysis reactions. 28089449 3 58 theme active 839:844 arg1 change 866:871 active site conformational change 839:871 active site conformational change 839:871 We show that substrate binding to non-catalytic domains and a conformationally stable active site promote CA transglycosylation, whereas a distinct pattern of active site conformational change is associated with CA hydrolysis. 28666223 7 0 theme PNGase 1034:1039 arg1 F 1041:1041 PNGase F 1034:1041 PNGase F 1034:1041 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 7 1 theme tryptic 946:952 arg1 reduction 994:1002 reduction 994:1002 reduction 994:1002 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 7 1 theme tryptic 946:952 arg1 IdeS 967:970 IdeS 967:970 IdeS 967:970 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 7 1 theme tryptic 946:952 arg1 digestion 983:991 papain digestion 976:991 papain digestion 976:991 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 7 1 theme tryptic 946:952 arg1 proteolysis 954:964 tryptic proteolysis 946:964 tryptic proteolysis 946:964 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 3 2 theme thanks 377:382 arg1 sample 369:374 the sample 365:374 the sample 365:374 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 5 3 theme biopharmaceutics 706:721 arg1 analysis 694:701 Structural elucidation and quality control (QC) analysis 646:701 Structural elucidation and quality control (QC) analysis of biopharmaceutics 646:721 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 5 4 theme elucidation 657:667 arg1 analysis 694:701 Structural elucidation and quality control (QC) analysis 646:701 Structural elucidation and quality control (QC) analysis of biopharmaceutics 646:721 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 8 5 theme modern 1083:1088 arg1 methods 1109:1115 Both historical and modern sample preparation methods 1063:1115 methods 1109:1115 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 4 6 theme spectrometric 622:634 arg1 analysis 636:643 chromatographic and mass spectrometric analysis 597:643 chromatographic and mass spectrometric analysis 597:643 These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. 28666223 3 7 theme sources 402:408 arg1 removal 391:397 the removal 387:397 the removal of sources of heterogeneity (e.g. N-glycans) 387:442 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 8 8 theme sample 1090:1095 arg1 methods 1109:1115 Both historical and modern sample preparation methods 1063:1115 methods 1109:1115 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 9 9 theme described 1305:1313 arg1 protocols 1315:1323 The described protocols 1301:1323 The described protocols 1301:1323 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 8 10 theme approved 1217:1224 arg1 trastuzumab 1165:1175 trastuzumab 1165:1175 trastuzumab 1165:1175 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 8 10 theme approved 1217:1224 arg1 products 1208:1215 two reference therapeutic mAb products 1178:1215 two reference therapeutic mAb products approved by Food and Drug Administration (FDA) 1178:1262 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 2 11 theme monoclonal 179:188 arg1 antibodies 190:199 therapeutic monoclonal antibodies 167:199 therapeutic monoclonal antibodies 167:199 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 3 12 dep N-glycans 433:441 arg1 e.g. 428:431 e.g. 428:431 e.g. 428:431 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 5 13 theme quality 673:679 arg1 QC 690:691 QC 690:691 QC 690:691 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 5 13 theme quality 673:679 arg1 control 681:687 quality control 673:687 quality control (QC) 673:692 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 8 14 theme reference 1182:1190 arg1 trastuzumab 1165:1175 trastuzumab 1165:1175 trastuzumab 1165:1175 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 8 14 theme reference 1182:1190 arg1 products 1208:1215 two reference therapeutic mAb products 1178:1215 two reference therapeutic mAb products approved by Food and Drug Administration (FDA) 1178:1262 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 2 15 theme therapeutic 167:177 arg1 antibodies 190:199 therapeutic monoclonal antibodies 167:199 therapeutic monoclonal antibodies 167:199 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 5 16 theme control 681:687 arg1 analysis 694:701 Structural elucidation and quality control (QC) analysis 646:701 Structural elucidation and quality control (QC) analysis of biopharmaceutics 646:721 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 9 17 theme preparation 1361:1371 arg1 methods 1373:1379 sample preparation methods 1354:1379 sample preparation methods 1354:1379 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 3 18 theme qualitative 310:320 arg1 information 322:332 quantitative and qualitative information 293:332 quantitative and qualitative information 293:332 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 3 19 dep enhanced 341:348 arg1 simplifying 353:363 simplifying 353:363 simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) 353:442 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 3 19 dep enhanced 341:348 arg1 decreasing 454:463 decreasing 454:463 decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation 454:543 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 1 20 theme Enzymatic 89:97 arg1 preparation 119:129 Enzymatic and chemical sample preparation 89:129 Enzymatic and chemical sample preparation 89:129 II - Enzymatic and chemical sample preparation. 28666223 9 21 theme sample 1354:1359 arg1 methods 1373:1379 sample preparation methods 1354:1379 sample preparation methods 1354:1379 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 8 22 theme Drug 1238:1241 arg1 FDA 1259:1261 FDA 1259:1261 FDA 1259:1261 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 8 22 theme Drug 1238:1241 arg1 Administration 1243:1256 Drug Administration 1238:1256 Drug Administration (FDA) 1238:1262 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 4 23 theme chromatographic 597:611 arg1 analysis 636:643 chromatographic and mass spectrometric analysis 597:643 chromatographic and mass spectrometric analysis 597:643 These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. 28666223 3 24 theme quantitative 293:304 arg1 information 322:332 quantitative and qualitative information 293:332 quantitative and qualitative information 293:332 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 7 25 theme papain 976:981 arg1 digestion 983:991 papain digestion 976:991 papain digestion 976:991 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 7 25 theme papain 976:981 arg1 proteolysis 954:964 tryptic proteolysis 946:964 tryptic proteolysis 946:964 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 3 26 theme molecular 469:477 arg1 size 479:482 the molecular size 465:482 the molecular size of the tested protein 465:504 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 3 27 theme heterogeneity 413:425 arg1 sources 402:408 sources 402:408 sources of heterogeneity (e.g. N-glycans) 402:442 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 5 28 theme subunit 756:762 arg1 levels 776:781 intact, subunit and peptide levels 748:781 intact, subunit and peptide levels 748:781 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 5 29 theme Structural 646:655 arg1 analysis 694:701 Structural elucidation and quality control (QC) analysis 646:701 Structural elucidation and quality control (QC) analysis of biopharmaceutics 646:721 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 1 30 theme chemical 103:110 arg1 preparation 119:129 Enzymatic and chemical sample preparation 89:129 Enzymatic and chemical sample preparation 89:129 II - Enzymatic and chemical sample preparation. 28666223 3 31 theme protein 498:504 arg1 size 479:482 the molecular size 465:482 the molecular size of the tested protein 465:504 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 4 32 theme mass 617:620 arg1 analysis 636:643 chromatographic and mass spectrometric analysis 597:643 chromatographic and mass spectrometric analysis 597:643 These approaches make the sample more suitable for chromatographic and mass spectrometric analysis. 28666223 7 33 theme EndoS2 1047:1052 arg1 enzymes 1054:1060 EndoS2 enzymes 1047:1060 EndoS2 enzymes 1047:1060 Protocols are described to perform tryptic proteolysis, IdeS and papain digestion, reduction as well as deglycosylation by PNGase F and EndoS2 enzymes. 28666223 2 34 theme proteins 213:220 arg1 characterization 147:162 The analytical characterization 132:162 The analytical characterization of therapeutic monoclonal antibodies and related proteins 132:220 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 1 35 theme sample 112:117 arg1 preparation 119:129 Enzymatic and chemical sample preparation 89:129 Enzymatic and chemical sample preparation 89:129 II - Enzymatic and chemical sample preparation. 28666223 5 36 theme peptide 768:774 arg1 levels 776:781 intact, subunit and peptide levels 748:781 intact, subunit and peptide levels 748:781 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 2 37 theme related 205:211 arg1 proteins 213:220 related proteins 205:220 related proteins 205:220 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 9 38 theme therapeutic 1397:1407 arg1 analysis 1417:1424 therapeutic protein analysis 1397:1424 therapeutic protein analysis 1397:1424 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 8 39 theme mAb 1204:1206 arg1 trastuzumab 1165:1175 trastuzumab 1165:1175 trastuzumab 1165:1175 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 8 39 theme mAb 1204:1206 arg1 products 1208:1215 two reference therapeutic mAb products 1178:1215 two reference therapeutic mAb products approved by Food and Drug Administration (FDA) 1178:1262 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 0 40 theme analytical 18:27 arg1 characterization 29:44 the analytical characterization 14:44 the analytical characterization of therapeutic monoclonal antibodies 14:81 Protocols for the analytical characterization of therapeutic monoclonal antibodies. 28666223 6 41 theme preparation 814:824 arg1 approaches 826:835 general sample preparation approaches 799:835 general sample preparation approaches used to attain peptide, subunit and glycan level analysis 799:893 In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. 28666223 3 42 theme tested 491:496 arg1 protein 498:504 the tested protein 487:504 the tested protein 487:504 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 0 43 theme monoclonal 61:70 arg1 antibodies 72:81 therapeutic monoclonal antibodies 49:81 therapeutic monoclonal antibodies 49:81 Protocols for the analytical characterization of therapeutic monoclonal antibodies. 28666223 9 44 theme protein 1409:1415 arg1 analysis 1417:1424 therapeutic protein analysis 1397:1424 therapeutic protein analysis 1397:1424 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 2 45 theme antibodies 190:199 arg1 characterization 147:162 The analytical characterization 132:162 The analytical characterization of therapeutic monoclonal antibodies and related proteins 132:220 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 6 46 theme sample 807:812 arg1 preparation 814:824 general sample preparation 799:824 general sample preparation approaches used to attain peptide, subunit and glycan level analysis 799:893 In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. 28666223 3 47 theme chemical 522:529 arg1 fragmentation 531:543 enzymatic or chemical fragmentation 509:543 enzymatic or chemical fragmentation 509:543 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 0 48 theme therapeutic 49:59 arg1 antibodies 72:81 therapeutic monoclonal antibodies 49:81 therapeutic monoclonal antibodies 49:81 Protocols for the analytical characterization of therapeutic monoclonal antibodies. 28666223 9 49 theme analysis 1417:1424 arg1 field 1388:1392 the field 1384:1392 the field of therapeutic protein analysis 1384:1424 The described protocols may help analysts to develop sample preparation methods in the field of therapeutic protein analysis. 28666223 2 50 theme sample 251:256 arg1 methodologies 270:282 various sample preparation methodologies 243:282 various sample preparation methodologies 243:282 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 2 51 theme preparation 258:268 arg1 methodologies 270:282 various sample preparation methodologies 243:282 various sample preparation methodologies 243:282 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 6 52 theme general 799:805 arg1 preparation 814:824 general sample preparation 799:824 general sample preparation approaches used to attain peptide, subunit and glycan level analysis 799:893 In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. 28666223 8 53 theme preparation 1097:1107 arg1 methods 1109:1115 Both historical and modern sample preparation methods 1063:1115 methods 1109:1115 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 5 54 theme intact 748:753 arg1 levels 776:781 intact, subunit and peptide levels 748:781 intact, subunit and peptide levels 748:781 Structural elucidation and quality control (QC) analysis of biopharmaceutics are usually performed at intact, subunit and peptide levels. 28666223 2 55 theme various 243:249 arg1 methodologies 270:282 various sample preparation methodologies 243:282 various sample preparation methodologies 243:282 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 28666223 3 56 theme enzymatic 509:517 arg1 fragmentation 531:543 enzymatic or chemical fragmentation 509:543 enzymatic or chemical fragmentation 509:543 Indeed, quantitative and qualitative information can be enhanced by simplifying the sample, thanks to the removal of sources of heterogeneity (e.g. N-glycans) and/or by decreasing the molecular size of the tested protein by enzymatic or chemical fragmentation. 28666223 6 57 theme level 880:884 arg1 analysis 886:893 glycan level analysis 873:893 glycan level analysis 873:893 In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. 28666223 8 58 theme therapeutic 1192:1202 arg1 trastuzumab 1165:1175 trastuzumab 1165:1175 trastuzumab 1165:1175 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 8 58 theme therapeutic 1192:1202 arg1 products 1208:1215 two reference therapeutic mAb products 1178:1215 two reference therapeutic mAb products approved by Food and Drug Administration (FDA) 1178:1262 Both historical and modern sample preparation methods were compared and evaluated using rituximab and trastuzumab, two reference therapeutic mAb products approved by Food and Drug Administration (FDA) and European Medicines Agency (EMA). 28666223 6 59 theme glycan 873:878 arg1 analysis 886:893 glycan level analysis 873:893 glycan level analysis 873:893 In this paper, general sample preparation approaches used to attain peptide, subunit and glycan level analysis are overviewed. 28666223 0 60 theme antibodies 72:81 arg1 characterization 29:44 the analytical characterization 14:44 the analytical characterization of therapeutic monoclonal antibodies 14:81 Protocols for the analytical characterization of therapeutic monoclonal antibodies. 28666223 2 61 theme analytical 136:145 arg1 characterization 147:162 The analytical characterization 132:162 The analytical characterization of therapeutic monoclonal antibodies and related proteins 132:220 The analytical characterization of therapeutic monoclonal antibodies and related proteins usually incorporates various sample preparation methodologies. 25709024 3 0 theme oxidized 368:375 arg1 MW 386:387 MW 47000 386:393 MW 47000 386:393 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 0 theme oxidized 368:375 arg1 dextran 377:383 oxidized dextran 368:383 oxidized dextran (MW 47000) 368:394 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 1 theme D 679:679 arg1 time 707:710 decimal reduction time 689:710 decimal reduction time 689:710 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 1 theme D 679:679 arg1 values 681:686 higher D values 672:686 higher D values (decimal reduction time) 672:711 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 4 2 theme CGTase 848:853 arg1 inactivation 810:821 irreversible inactivation 797:821 irreversible inactivation of native and conjugated CGTase 797:853 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 5 3 dep ΔG 1199:1200 arg1 energy 1170:1175 free energy 1165:1175 free energy of substrate binding 1165:1196 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 4 theme native 506:511 arg1 enzyme 513:518 the native enzyme 502:518 the native enzyme 502:518 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 5 theme activation 1065:1074 arg1 E-T 1156:1158 (free energy of transition state) ΔG*E-T 1119:1158 (free energy of transition state) ΔG*E-T 1119:1158 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 5 theme activation 1065:1074 arg1 E-S 1202:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 5 theme activation 1065:1074 arg1 enthalpy 1053:1060 The enthalpy 1049:1060 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*) 1049:1116 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 6 theme free 1120:1123 arg1 energy 1125:1130 free energy 1120:1130 free energy of transition state 1120:1150 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 7 theme free 1086:1089 arg1 energy 1091:1096 free energy 1086:1096 free energy 1086:1096 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 4 8 theme conjugated 837:846 arg1 CGTase 848:853 native and conjugated CGTase 826:853 native and conjugated CGTase 826:853 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 3 9 theme deactivation 622:633 arg1 kd 650:651 kd 650:651 kd 650:651 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 9 theme deactivation 622:633 arg1 rate 644:647 lower deactivation constant rate 616:647 lower deactivation constant rate (kd) 616:652 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 10 theme thermodynamic 971:983 arg1 analysis 985:992 thermodynamic analysis 971:992 thermodynamic analysis for cyclodextrin production from starch 971:1032 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 4 11 theme irreversible 797:808 arg1 inactivation 810:821 irreversible inactivation 797:821 irreversible inactivation of native and conjugated CGTase 797:853 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 5 12 theme free 1165:1168 arg1 energy 1170:1175 free energy 1165:1175 free energy of substrate binding 1165:1196 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 13 theme decimal 689:695 arg1 time 707:710 decimal reduction time 689:710 decimal reduction time 689:710 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 13 theme decimal 689:695 arg1 values 681:686 higher D values 672:686 higher D values (decimal reduction time) 672:711 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 14 theme higher 558:563 arg1 temperature 573:583 higher optimum temperature 558:583 higher optimum temperature 558:583 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 15 theme transition 1135:1144 arg1 state 1146:1150 transition state 1135:1150 transition state 1135:1150 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 4 16 theme deactivation 936:947 arg1 entropy 906:912 entropy 906:912 entropy (ΔS*) 906:918 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 4 16 theme deactivation 936:947 arg1 * 917:917 ΔS* 915:917 ΔS* 915:917 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 4 16 theme deactivation 936:947 arg1 enthalpy 924:931 enthalpy 924:931 enthalpy of deactivation (ΔH*) 924:953 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 3 17 theme reduction 697:705 arg1 time 707:710 decimal reduction time 689:710 decimal reduction time 689:710 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 17 theme reduction 697:705 arg1 values 681:686 higher D values 672:686 higher D values (decimal reduction time) 672:711 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 2 18 theme thermodynamic 316:328 arg1 stability 330:338 kinetic and thermodynamic stability 304:338 kinetic and thermodynamic stability 304:338 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 2 18 theme thermodynamic 316:328 arg1 activity 294:301 the retained activity 281:301 the retained activity 281:301 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 3 19 theme highest 456:462 arg1 T1/2 475:478 T1/2 475:478 T1/2 475:478 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 19 theme highest 456:462 arg1 half-life 464:472 the highest half-life 452:472 the highest half-life (T1/2) 452:479 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 6 20 theme significant 1272:1282 arg1 impact 1284:1289 significant impact 1272:1289 significant impact 1272:1289 Similarly, there was significant impact on improvement of kcat, kcat/Km values. 25709024 1 21 theme Cyclodextrin 105:116 arg1 CGTase 139:144 CGTase 139:144 CGTase 139:144 Cyclodextrin glycosyltransferase (CGTase) was covalently coupled to five oxidized polysaccharides differing in structure and chemical nature. 25709024 1 21 theme Cyclodextrin 105:116 arg1 glycosyltransferase 118:136 Cyclodextrin glycosyltransferase 105:136 Cyclodextrin glycosyltransferase (CGTase) 105:145 Cyclodextrin glycosyltransferase (CGTase) was covalently coupled to five oxidized polysaccharides differing in structure and chemical nature. 25709024 3 22 dep energy 603:608 arg1 range 736:740 the temperature range 720:740 the temperature range of 60-80°C 720:751 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 23 theme specific 421:428 arg1 activity 430:437 the highest retained specific activity 400:437 the highest retained specific activity (70.05%) 400:446 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 23 theme specific 421:428 arg1 % 445:445 70.05% 440:445 70.05% 440:445 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 24 theme activation 1101:1110 arg1 E-T 1156:1158 (free energy of transition state) ΔG*E-T 1119:1158 (free energy of transition state) ΔG*E-T 1119:1158 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 24 theme activation 1101:1110 arg1 E-S 1202:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 24 theme activation 1101:1110 arg1 enthalpy 1053:1060 The enthalpy 1049:1060 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*) 1049:1116 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 25 from starch 1027:1032 arg1 production 1011:1020 cyclodextrin production 998:1020 cyclodextrin production from starch 998:1032 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 7 26 theme native 1336:1341 arg1 enzyme 1358:1363 Both native and conjugated enzyme 1331:1363 Both native and conjugated enzyme 1331:1363 Both native and conjugated enzyme produce α-cyclodextrin from starch. 25709024 3 27 with CGTase 356:361 arg1 MW 386:387 MW 47000 386:393 MW 47000 386:393 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 27 with CGTase 356:361 arg1 dextran 377:383 oxidized dextran 368:383 oxidized dextran (MW 47000) 368:394 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 0 28 theme Catalytic 0:8 arg1 properties 28:37 Catalytic and thermodynamic properties 0:37 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase 0:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 4 29 theme native 826:831 arg1 CGTase 848:853 native and conjugated CGTase 826:853 native and conjugated CGTase 826:853 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 5 30 dep E-T 1156:1158 arg1 energy 1125:1130 free energy 1120:1130 free energy of transition state 1120:1150 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 31 theme lower 586:590 arg1 Ea 611:612 Ea 611:612 Ea 611:612 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 31 theme lower 586:590 arg1 energy 603:608 lower activation energy 586:608 lower activation energy (Ea) 586:613 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 1 32 theme chemical 230:237 arg1 nature 239:244 chemical nature 230:244 chemical nature 230:244 Cyclodextrin glycosyltransferase (CGTase) was covalently coupled to five oxidized polysaccharides differing in structure and chemical nature. 25709024 3 33 theme highest 404:410 arg1 activity 430:437 the highest retained specific activity 400:437 the highest retained specific activity (70.05%) 400:446 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 33 theme highest 404:410 arg1 % 445:445 70.05% 440:445 70.05% 440:445 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 34 theme optimum 565:571 arg1 temperature 573:583 higher optimum temperature 558:583 higher optimum temperature 558:583 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 35 theme ΔG 1199:1200 arg1 E-S 1202:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 0 36 theme thermodynamic 14:26 arg1 properties 28:37 Catalytic and thermodynamic properties 0:37 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase 0:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 3 37 theme conjugated 525:534 arg1 preparation 536:546 the conjugated preparation 521:546 the conjugated preparation 521:546 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 38 theme * 1201:1201 arg1 E-S 1202:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 39 theme retained 412:419 arg1 activity 430:437 the highest retained specific activity 400:437 the highest retained specific activity (70.05%) 400:446 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 39 theme retained 412:419 arg1 % 445:445 70.05% 440:445 70.05% 440:445 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 0 40 theme glycosylated 42:53 arg1 glycosyltransferase 84:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 6 41 theme values 1323:1328 arg1 improvement 1294:1304 improvement 1294:1304 improvement of kcat, kcat/Km values 1294:1328 Similarly, there was significant impact on improvement of kcat, kcat/Km values. 25709024 4 42 theme parameters 782:791 arg1 values 758:763 The values 754:763 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase 754:853 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 5 43 theme E-S 1202:1204 arg1 values 1206:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 44 theme constant 635:642 arg1 kd 650:651 kd 650:651 kd 650:651 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 44 theme constant 635:642 arg1 rate 644:647 lower deactivation constant rate 616:647 lower deactivation constant rate (kd) 616:652 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 0 45 dep Bacillus 55:62 arg1 cereus 64:69 cereus 64:69 cereus 64:69 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 3 46 theme 60-80°C 745:751 arg1 range 736:740 the temperature range 720:740 the temperature range of 60-80°C 720:751 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 47 theme activation 592:601 arg1 Ea 611:612 Ea 611:612 Ea 611:612 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 47 theme activation 592:601 arg1 energy 603:608 lower activation energy 586:608 lower activation energy (Ea) 586:613 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 6 48 theme kcat/Km 1315:1321 arg1 values 1323:1328 kcat, kcat/Km values 1309:1328 values 1323:1328 Similarly, there was significant impact on improvement of kcat, kcat/Km values. 25709024 0 49 theme Bacillus 55:62 arg1 glycosyltransferase 84:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 3 50 theme temperature 724:734 arg1 range 736:740 the temperature range 720:740 the temperature range of 60-80°C 720:751 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 51 theme higher 655:660 arg1 T1/2 662:665 higher T1/2 655:665 higher T1/2 655:665 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 5 52 theme cyclodextrin 998:1009 arg1 production 1011:1020 cyclodextrin production 998:1020 cyclodextrin production from starch 998:1032 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 53 theme enthalpy 1053:1060 arg1 values 1206:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 4 54 theme thermodynamic 768:780 arg1 parameters 782:791 thermodynamic parameters 768:791 thermodynamic parameters 768:791 The values of thermodynamic parameters for irreversible inactivation of native and conjugated CGTase indicated that conjugation significantly decreased entropy (ΔS*) and enthalpy of deactivation (ΔH*). 25709024 5 55 theme analysis 985:992 arg1 results 960:966 The results 956:966 The results of thermodynamic analysis for cyclodextrin production from starch 956:1032 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 56 theme lower 616:620 arg1 kd 650:651 kd 650:651 kd 650:651 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 56 theme lower 616:620 arg1 rate 644:647 lower deactivation constant rate 616:647 lower deactivation constant rate (kd) 616:652 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 2 57 theme kinetic 304:310 arg1 stability 330:338 kinetic and thermodynamic stability 304:338 kinetic and thermodynamic stability 304:338 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 2 57 theme kinetic 304:310 arg1 activity 294:301 the retained activity 281:301 the retained activity 281:301 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 3 58 contain had 396:398 arg2 activity 430:437 the highest retained specific activity 400:437 the highest retained specific activity (70.05%) 400:446 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 58 contain had 396:398 arg2 T1/2 475:478 T1/2 475:478 T1/2 475:478 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 58 contain had 396:398 arg2 half-life 464:472 the highest half-life 452:472 the highest half-life (T1/2) 452:479 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 58 contain had 396:398 arg1 CGTase 356:361 The conjugated CGTase 341:361 The conjugated CGTase with oxidized dextran (MW 47000) 341:394 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 58 contain had 396:398 arg2 % 445:445 70.05% 440:445 70.05% 440:445 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 7 59 theme conjugated 1347:1356 arg1 enzyme 1358:1363 Both native and conjugated enzyme 1331:1363 Both native and conjugated enzyme 1331:1363 Both native and conjugated enzyme produce α-cyclodextrin from starch. 25709024 5 60 theme state 1146:1150 arg1 energy 1125:1130 free energy 1120:1130 free energy of transition state 1120:1150 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 3 61 theme conjugated 345:354 arg1 CGTase 356:361 The conjugated CGTase 341:361 The conjugated CGTase with oxidized dextran (MW 47000) 341:394 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 0 62 gly glycosylated 42:53 arg1 glycosyltransferase 84:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 5 63 theme substrate 1180:1188 arg1 binding 1190:1196 substrate binding 1180:1196 substrate binding 1180:1196 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 0 64 theme glycosyltransferase 84:102 arg1 properties 28:37 Catalytic and thermodynamic properties 0:37 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase 0:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 6 65 theme kcat 1309:1312 arg1 values 1323:1328 kcat, kcat/Km values 1309:1328 values 1323:1328 Similarly, there was significant impact on improvement of kcat, kcat/Km values. 25709024 5 66 theme energy 1091:1096 arg1 E-T 1156:1158 (free energy of transition state) ΔG*E-T 1119:1158 (free energy of transition state) ΔG*E-T 1119:1158 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 66 theme energy 1091:1096 arg1 E-S 1202:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 (free energy of substrate binding) ΔG*E-S 1164:1204 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 66 theme energy 1091:1096 arg1 enthalpy 1053:1060 The enthalpy 1049:1060 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*) 1049:1116 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 67 theme conjugated 1232:1241 arg1 CGTase 1243:1248 the conjugated CGTase 1228:1248 the conjugated CGTase 1228:1248 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 68 theme binding 1190:1196 arg1 energy 1170:1175 free energy 1165:1175 free energy of substrate binding 1165:1196 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 69 theme ΔG 1153:1154 arg1 E-T 1156:1158 (free energy of transition state) ΔG*E-T 1119:1158 (free energy of transition state) ΔG*E-T 1119:1158 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 0 70 theme cyclodextrin 71:82 arg1 glycosyltransferase 84:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 glycosylated Bacillus cereus cyclodextrin glycosyltransferase 42:102 Catalytic and thermodynamic properties of glycosylated Bacillus cereus cyclodextrin glycosyltransferase. 25709024 3 71 theme higher 672:677 arg1 time 707:710 decimal reduction time 689:710 decimal reduction time 689:710 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 3 71 theme higher 672:677 arg1 values 681:686 higher D values 672:686 higher D values (decimal reduction time) 672:711 The conjugated CGTase with oxidized dextran (MW 47000) had the highest retained specific activity (70.05%) and the highest half-life (T1/2) at 80°C. Compared to the native enzyme, the conjugated preparation exhibited higher optimum temperature, lower activation energy (Ea), lower deactivation constant rate (kd), higher T1/2, and higher D values (decimal reduction time) within the temperature range of 60-80°C. 25709024 2 72 theme retained 285:292 arg1 stability 330:338 kinetic and thermodynamic stability 304:338 kinetic and thermodynamic stability 304:338 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 2 72 theme retained 285:292 arg1 activity 294:301 the retained activity 281:301 the retained activity 281:301 The conjugates were evaluated for the retained activity, kinetic and thermodynamic stability. 25709024 1 73 theme oxidized 178:185 arg1 polysaccharides 187:201 five oxidized polysaccharides 173:201 five oxidized polysaccharides differing in structure and chemical nature 173:244 Cyclodextrin glycosyltransferase (CGTase) was covalently coupled to five oxidized polysaccharides differing in structure and chemical nature. 25709024 5 74 theme E-T 1156:1158 arg1 values 1206:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values 1049:1211 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 25709024 5 75 theme * 1155:1155 arg1 E-T 1156:1158 (free energy of transition state) ΔG*E-T 1119:1158 (free energy of transition state) ΔG*E-T 1119:1158 The results of thermodynamic analysis for cyclodextrin production from starch indicated that The enthalpy of activation (ΔH*) and free energy of activation (ΔG*), (free energy of transition state) ΔG*E-T and (free energy of substrate binding) ΔG*E-S values were lower for the conjugated CGTase. 24715520 3 0 theme two-step 544:551 arg1 methodology 553:563 a two-step methodology 542:563 a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays 542:684 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 6 1 theme multivalent 1119:1129 arg1 glycoconjugates 1131:1145 multivalent glycoconjugates 1119:1145 multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics 1119:1206 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 5 2 theme chemical 972:979 arg1 handle 981:986 a chemical handle 970:986 a chemical handle for further modification 970:1011 The terminal iodide served as a chemical handle for further modification. 24715520 5 2 theme chemical 972:979 arg1 iodide 953:958 The terminal iodide 940:958 The terminal iodide 940:958 The terminal iodide served as a chemical handle for further modification. 24715520 4 3 theme glycosyl 774:781 arg1 iodides 783:789 glycosyl iodides 774:789 glycosyl iodides 774:789 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 3 4 theme per-O-acetylated 588:603 arg1 oligosaccharides 605:620 per-O-acetylated oligosaccharides 588:620 per-O-acetylated oligosaccharides 588:620 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 3 5 theme functionalized 625:638 arg1 linkers 640:646 functionalized linkers 625:646 functionalized linkers 625:646 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 1 6 theme biological 297:306 arg1 roles 308:312 the biological roles 293:312 the biological roles of carbohydrates 293:329 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 0 7 theme applications 100:111 arg1 functionalization 9:25 Two-step functionalization 0:25 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 0:143 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 6 8 theme azide-alkyne 1083:1094 arg1 cycloaddition 1096:1108 copper-catalyzed azide-alkyne cycloaddition 1066:1108 copper-catalyzed azide-alkyne cycloaddition 1066:1108 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 3 9 theme oligosaccharides 605:620 arg1 conjugation 573:583 the conjugation 569:583 the conjugation of per-O-acetylated oligosaccharides to functionalized linkers 569:646 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 2 10 theme synthetic 363:371 arg1 technologies 373:384 many modern isolation and synthetic technologies 337:384 technologies 373:384 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 4 11 theme single 927:932 arg1 step 934:937 a single step 925:937 a single step 925:937 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 4 12 theme commercial 737:746 arg1 sources 748:754 both synthetic and commercial sources 718:754 both synthetic and commercial sources 718:754 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 0 13 theme multivalent 116:126 arg1 glycoconjugates 128:142 multivalent glycoconjugates 116:142 multivalent glycoconjugates 116:142 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 6 14 theme Gb3 1150:1152 arg1 glycoconjugates 1131:1145 multivalent glycoconjugates 1119:1145 multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics 1119:1206 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 2 15 theme sugars 429:434 arg1 sugars 429:434 free sugars 424:434 free sugars 424:434 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 2 15 theme sugars 429:434 arg1 variety 413:419 a wide variety 406:419 a wide variety of free sugars 406:434 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 2 16 theme free 424:427 arg1 sugars 429:434 free sugars 424:434 free sugars 424:434 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 0 17 theme Two-step 0:7 arg1 functionalization 9:25 Two-step functionalization 0:25 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 0:143 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 6 18 theme copper-catalyzed 1066:1081 arg1 cycloaddition 1096:1108 copper-catalyzed azide-alkyne cycloaddition 1066:1108 copper-catalyzed azide-alkyne cycloaddition 1066:1108 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 1 19 theme potential 216:224 arg1 conjugates 161:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 19 theme potential 216:224 arg1 glycolipids 199:209 glycolipids 199:209 glycolipids 199:209 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 19 theme potential 216:224 arg1 glycoproteins 181:193 glycoproteins 181:193 glycoproteins 181:193 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 19 theme potential 216:224 arg1 chemotherapeutics 226:242 potential chemotherapeutics 216:242 potential chemotherapeutics 216:242 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 0 20 theme oligosaccharides 30:45 arg1 functionalization 9:25 Two-step functionalization 0:25 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 0:143 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 1 21 theme carbohydrates 317:329 arg1 roles 308:312 the biological roles 293:312 the biological roles of carbohydrates 293:329 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 2 22 theme modern 342:347 arg1 isolation 349:357 many modern isolation and synthetic technologies 337:384 isolation 349:357 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 3 23 dep various 669:675 arg1 displays 677:684 displays 677:684 displays 677:684 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 2 24 theme many 337:340 arg1 isolation 349:357 many modern isolation and synthetic technologies 337:384 isolation 349:357 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 2 25 theme increasing 446:455 arg1 need 457:460 increasing need 446:460 increasing need for general methodologies for carbohydrate functionalization 446:521 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 0 26 theme glycosyl 53:60 arg1 iodide 62:67 glycosyl iodide 53:67 glycosyl iodide 53:67 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 5 27 theme further 992:998 arg1 modification 1000:1011 further modification 992:1011 further modification 992:1011 The terminal iodide served as a chemical handle for further modification. 24715520 4 28 theme iodopropyl 900:909 arg1 conjugates 911:920 iodopropyl conjugates 900:920 iodopropyl conjugates 900:920 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 4 29 theme synthetic 723:731 arg1 sources 748:754 both synthetic and commercial sources 718:754 both synthetic and commercial sources 718:754 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 6 30 theme corresponding 1034:1046 arg1 azide 1048:1052 the corresponding azide 1030:1052 the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition 1030:1108 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 2 31 theme general 466:472 arg1 methodologies 474:486 general methodologies 466:486 general methodologies 466:486 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 6 32 theme further 1158:1164 arg1 investigation 1166:1178 further investigation 1158:1178 further investigation as anti-cancer therapeutics 1158:1206 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 2 33 theme carbohydrate 492:503 arg1 functionalization 505:521 carbohydrate functionalization 492:521 carbohydrate functionalization 492:521 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 1 34 theme Oligosaccharide 145:159 arg1 conjugates 161:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 34 theme Oligosaccharide 145:159 arg1 tools 269:273 useful tools 262:273 useful tools for understanding the biological roles of carbohydrates 262:329 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 34 theme Oligosaccharide 145:159 arg1 glycolipids 199:209 glycolipids 199:209 glycolipids 199:209 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 34 theme Oligosaccharide 145:159 arg1 glycoproteins 181:193 glycoproteins 181:193 glycoproteins 181:193 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 34 theme Oligosaccharide 145:159 arg1 chemotherapeutics 226:242 potential chemotherapeutics 216:242 potential chemotherapeutics 216:242 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 4 35 theme trimethylene 873:884 arg1 oxide 886:890 trimethylene oxide 873:890 trimethylene oxide 873:890 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 1 36 gly glycoproteins 181:193 arg1 glycoproteins 181:193 glycoproteins 181:193 glycoproteins 181:193 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 4 37 theme reactive 821:828 arg1 donors 830:835 reactive donors 821:835 reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step 821:937 Oligosaccharides obtained from both synthetic and commercial sources were converted to glycosyl iodides and activated with I2 to form reactive donors that were subsequently trapped with trimethylene oxide to form iodopropyl conjugates in a single step. 24715520 3 38 used used 660:663 arg2 methodology 553:563 a two-step methodology 542:563 a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays 542:684 Herein, we report a two-step methodology for the conjugation of per-O-acetylated oligosaccharides to functionalized linkers that can be used for various displays. 24715520 1 39 theme useful 262:267 arg1 tools 269:273 useful tools 262:273 useful tools for understanding the biological roles of carbohydrates 262:329 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 39 theme useful 262:267 arg1 conjugates 161:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates 145:170 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 39 theme useful 262:267 arg1 glycolipids 199:209 glycolipids 199:209 glycolipids 199:209 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 1 39 theme useful 262:267 arg1 glycoproteins 181:193 glycoproteins 181:193 glycoproteins 181:193 Oligosaccharide conjugates, such as glycoproteins and glycolipids, are potential chemotherapeutics and also serve as useful tools for understanding the biological roles of carbohydrates. 24715520 6 40 theme anti-cancer 1183:1193 arg1 therapeutics 1195:1206 anti-cancer therapeutics 1183:1206 anti-cancer therapeutics 1183:1206 Conversion into the corresponding azide followed by copper-catalyzed azide-alkyne cycloaddition afforded multivalent glycoconjugates of Gb3 for further investigation as anti-cancer therapeutics. 24715520 5 41 theme terminal 944:951 arg1 handle 981:986 a chemical handle 970:986 a chemical handle for further modification 970:1011 The terminal iodide served as a chemical handle for further modification. 24715520 5 41 theme terminal 944:951 arg1 iodide 953:958 The terminal iodide 940:958 The terminal iodide 940:958 The terminal iodide served as a chemical handle for further modification. 24715520 0 42 theme trimethylene 73:84 arg1 oxide 86:90 trimethylene oxide 73:90 trimethylene oxide 73:90 Two-step functionalization of oligosaccharides using glycosyl iodide and trimethylene oxide and its applications to multivalent glycoconjugates. 24715520 2 43 theme wide 408:411 arg1 sugars 429:434 free sugars 424:434 free sugars 424:434 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 24715520 2 43 theme wide 408:411 arg1 variety 413:419 a wide variety 406:419 a wide variety of free sugars 406:434 With many modern isolation and synthetic technologies providing access to a wide variety of free sugars, there is increasing need for general methodologies for carbohydrate functionalization. 25275130 14 0 theme potential 2389:2397 arg1 strategy 2399:2406 a potential strategy 2387:2406 a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1 2387:2480 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 6 1 with viruses 838:844 arg1 glycans 890:896 more complex Env glycans 873:896 more complex Env glycans 873:896 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 1 with viruses 838:844 arg1 oligomannose 856:867 less oligomannose 851:867 less oligomannose 851:867 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 8 2 theme crucial 1516:1522 arg1 role 1524:1527 a crucial role 1514:1527 a crucial role 1514:1527 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 14 3 theme Env 2291:2293 arg1 moieties 2302:2309 Env glycan moieties 2291:2309 Env glycan moieties 2291:2309 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 3 4 theme glycosylation 471:483 arg1 Studies 454:460 Studies 454:460 Studies of HIV-1 glycosylation 454:483 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 6 5 dep lymphocytes 1166:1176 arg1 iv 1139:1140 iv 1139:1140 iv 1139:1140 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 5 6 with viruses 676:682 arg1 content 709:715 modified Env glycan content 689:715 modified Env glycan content 689:715 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 5 6 with viruses 676:682 arg1 heterogeneity 721:733 heterogeneity 721:733 heterogeneity 721:733 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 6 7 with viruses 899:905 arg1 oligomannose 917:928 more oligomannose 912:928 more oligomannose 912:928 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 8 theme primary 1158:1164 arg1 lymphocytes 1166:1176 (iv) trans-infected primary lymphocytes 1138:1176 (iv) trans-infected primary lymphocytes 1138:1176 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 11 9 theme HIV-1 1929:1933 arg1 glycosylation 1935:1947 HIV-1 glycosylation 1929:1947 HIV-1 glycosylation 1929:1947 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 14 10 theme glycan 2364:2369 arg1 moieties 2371:2378 Env glycan moieties 2360:2378 Env glycan moieties 2360:2378 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 7 11 theme primary 1307:1313 arg1 lymphocytes 1315:1325 primary lymphocytes 1307:1325 primary lymphocytes 1307:1325 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 4 12 theme moieties 602:609 arg1 role 583:586 the role 579:586 the role of Env glycan moieties on HIV-1 transmission 579:631 Also, the role of Env glycan moieties on HIV-1 transmission has not been systematically defined. 25275130 13 13 theme major 2230:2234 arg1 events 2236:2241 the major events 2226:2241 the major events of HIV-1 transmission 2226:2263 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 7 14 theme human 1338:1342 arg1 tissue 1355:1360 fresh human intestinal tissue 1332:1360 fresh human intestinal tissue 1332:1360 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 4 15 theme HIV-1 614:618 arg1 transmission 620:631 HIV-1 transmission 614:631 HIV-1 transmission 614:631 Also, the role of Env glycan moieties on HIV-1 transmission has not been systematically defined. 25275130 10 16 theme HIV-1 1687:1691 arg1 N-glycans 1697:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 1 17 theme molecular 163:171 arg1 mass 173:176 the Env molecular mass 155:176 the Env molecular mass 155:176 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 3 18 theme glycosylation 524:536 arg1 position 512:519 the position 508:519 the position of glycosylation 508:536 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 4 19 from role 583:586 arg1 transmission 620:631 HIV-1 transmission 614:631 HIV-1 transmission 614:631 Also, the role of Env glycan moieties on HIV-1 transmission has not been systematically defined. 25275130 13 20 theme discordant 2204:2213 arg1 effects 2215:2221 discordant effects 2204:2221 discordant effects on the major events of HIV-1 transmission 2204:2263 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 6 21 dep bound 1086:1090 arg1 iii 1081:1083 iii 1081:1083 iii 1081:1083 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 1 22 gly glycosylated 115:126 arg1 Env 99:101 Env 99:101 Env 99:101 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 1 22 gly glycosylated 115:126 arg1 protein 90:96 The HIV-1 envelope protein 71:96 UNLABELLED The HIV-1 envelope protein (Env) 60:102 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 2 23 theme HIV-1 210:214 arg1 N-glycans 220:228 HIV-1 Env N-glycans 210:228 HIV-1 Env N-glycans 210:228 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 5 24 theme major 790:794 arg1 events 796:801 the major events 786:801 the major events of HIV-1 transmission 786:823 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 6 25 theme Env 886:888 arg1 glycans 890:896 more complex Env glycans 873:896 more complex Env glycans 873:896 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 7 26 theme impaired 1264:1271 arg1 infectivity 1273:1283 impaired infectivity 1264:1283 impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue 1264:1360 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 11 27 theme glycosylation 1988:2000 arg1 position 1976:1983 the position 1972:1983 the position of glycosylation 1972:2000 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 13 28 theme N-linked 2175:2182 arg1 glycans 2188:2194 N-linked Env glycans 2175:2194 N-linked Env glycans 2175:2194 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 9 29 theme vaccine 1624:1630 arg1 strategies 1632:1641 vaccine strategies 1624:1641 vaccine strategies 1624:1641 Env glycosylation should be taken into consideration in the development of vaccine strategies to interdict HIV-1 transmission. 25275130 10 30 theme key 1744:1746 arg1 roles 1748:1752 key roles 1744:1752 key roles 1744:1752 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 12 31 theme Env 2119:2121 arg1 moieties 2130:2137 Env glycan moieties 2119:2137 Env glycan moieties 2119:2137 In the study described in this report, we investigated systematically the role of Env glycan moieties on HIV-1 transmission. 25275130 8 32 from structures 1488:1497 arg1 Env 1502:1504 Env 1502:1504 Env playing a crucial role in HIV-1 infection 1502:1546 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 6 33 theme dendritic 1109:1117 arg1 cells 1119:1123 monocyte-derived dendritic cells 1092:1123 monocyte-derived dendritic cells (MoDCs) 1092:1131 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 33 theme dendritic 1109:1117 arg1 MoDCs 1126:1130 MoDCs 1126:1130 MoDCs 1126:1130 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 5 34 theme glycan 767:772 arg1 moieties 774:781 Env glycan moieties 763:781 Env glycan moieties 763:781 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 9 35 theme HIV-1 1656:1660 arg1 transmission 1662:1673 HIV-1 transmission 1656:1673 HIV-1 transmission 1656:1673 Env glycosylation should be taken into consideration in the development of vaccine strategies to interdict HIV-1 transmission. 25275130 8 36 theme Env 1378:1380 arg1 glycans 1382:1388 N-linked Env glycans 1369:1388 N-linked Env glycans 1369:1388 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 14 37 theme moieties 2371:2378 arg1 modulation 2346:2355 modulation 2346:2355 modulation of Env glycan moieties 2346:2378 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 12 38 theme moieties 2130:2137 arg1 role 2111:2114 the role 2107:2114 the role of Env glycan moieties on HIV-1 transmission 2107:2159 In the study described in this report, we investigated systematically the role of Env glycan moieties on HIV-1 transmission. 25275130 12 39 theme HIV-1 2142:2146 arg1 transmission 2148:2159 HIV-1 transmission 2142:2159 HIV-1 transmission 2142:2159 In the study described in this report, we investigated systematically the role of Env glycan moieties on HIV-1 transmission. 25275130 14 40 theme prophylactic 2446:2457 arg1 vaccines 2459:2466 therapeutic or prophylactic vaccines 2431:2466 therapeutic or prophylactic vaccines against HIV-1 2431:2480 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 5 41 theme Env 698:700 arg1 content 709:715 modified Env glycan content 689:715 modified Env glycan content 689:715 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 5 42 from effects 752:758 arg1 events 796:801 the major events 786:801 the major events of HIV-1 transmission 786:823 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 1 43 theme envelope 81:88 arg1 Env 99:101 Env 99:101 Env 99:101 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 1 43 theme envelope 81:88 arg1 protein 90:96 The HIV-1 envelope protein 71:96 UNLABELLED The HIV-1 envelope protein (Env) 60:102 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 6 44 theme cell 1010:1013 arg1 monolayer 1015:1023 an epithelial cell monolayer 996:1023 an epithelial cell monolayer 996:1023 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 0 45 theme HIV-1 0:4 arg1 moieties 22:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties modulate HIV-1 transmission. 25275130 7 46 theme more 1211:1214 arg1 oligomannose 1216:1227 more oligomannose 1211:1227 more oligomannose 1211:1227 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 2 47 theme antibody 390:397 arg1 neutralization 399:412 antibody neutralization 390:412 antibody neutralization 390:412 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 6 48 dep monolayer 1015:1023 arg1 attached 1031:1038 attached 1031:1038 (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs 1026:1186 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 0 49 theme glycan 15:20 arg1 moieties 22:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties modulate HIV-1 transmission. 25275130 14 50 theme vaccines 2459:2466 arg1 development 2416:2426 the development 2412:2426 the development of therapeutic or prophylactic vaccines against HIV-1 2412:2480 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 2 51 theme receptor 436:443 arg1 binding 445:451 receptor binding 436:451 receptor binding 436:451 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 8 52 theme HIV-1 1440:1444 arg1 transmission 1446:1457 HIV-1 transmission 1440:1457 HIV-1 transmission 1440:1457 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 6 53 link monocyte-derived 1043:1058 arg1 MDMs 1073:1076 MDMs 1073:1076 MDMs 1073:1076 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 53 link monocyte-derived 1043:1058 arg1 macrophages 1060:1070 monocyte-derived macrophages 1043:1070 monocyte-derived macrophages (MDMs) 1043:1077 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 14 54 theme Env 2360:2362 arg1 moieties 2371:2378 Env glycan moieties 2360:2378 Env glycan moieties 2360:2378 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 2 55 theme Env 311:313 arg1 structure 315:323 Env structure 311:323 Env structure 311:323 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 8 56 from effects 1409:1415 arg1 events 1430:1435 the major events 1420:1435 the major events of HIV-1 transmission 1420:1457 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 6 57 theme monocyte-derived 1043:1058 arg1 MDMs 1073:1076 MDMs 1073:1076 MDMs 1073:1076 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 57 theme monocyte-derived 1043:1058 arg1 macrophages 1060:1070 monocyte-derived macrophages 1043:1070 monocyte-derived macrophages (MDMs) 1043:1077 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 8 58 theme mature 1465:1470 arg1 structures 1488:1497 mature oligosaccharide structures 1465:1497 mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection 1465:1546 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 10 59 theme surface 1787:1793 arg1 exposure 1795:1802 surface exposure 1787:1802 surface exposure 1787:1802 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 8 60 theme HIV-1 1532:1536 arg1 infection 1538:1546 HIV-1 infection 1532:1546 HIV-1 infection 1532:1546 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 13 61 theme transmission 2252:2263 arg1 events 2236:2241 the major events 2226:2241 the major events of HIV-1 transmission 2226:2263 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 12 62 from role 2111:2114 arg1 transmission 2148:2159 HIV-1 transmission 2142:2159 HIV-1 transmission 2142:2159 In the study described in this report, we investigated systematically the role of Env glycan moieties on HIV-1 transmission. 25275130 14 63 theme glycan 2295:2300 arg1 moieties 2302:2309 Env glycan moieties 2291:2309 Env glycan moieties 2291:2309 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 3 64 theme HIV-1 465:469 arg1 glycosylation 471:483 HIV-1 glycosylation 465:483 HIV-1 glycosylation 465:483 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 9 65 gly glycosylation 1553:1565 arg1 development 1609:1619 the development 1605:1619 the development of vaccine strategies to interdict HIV-1 transmission 1605:1673 Env glycosylation should be taken into consideration in the development of vaccine strategies to interdict HIV-1 transmission. 25275130 6 66 theme less 851:854 arg1 oligomannose 856:867 less oligomannose 851:867 less oligomannose 851:867 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 11 67 theme glycans 2028:2034 arg1 types 2019:2023 the types 2015:2023 the types of glycans 2015:2034 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 10 68 theme Env 1693:1695 arg1 N-glycans 1697:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 7 69 theme intestinal 1344:1353 arg1 tissue 1355:1360 fresh human intestinal tissue 1332:1360 fresh human intestinal tissue 1332:1360 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 9 70 theme Env 1549:1551 arg1 glycosylation 1553:1565 Env glycosylation 1549:1565 Env glycosylation 1549:1565 Env glycosylation should be taken into consideration in the development of vaccine strategies to interdict HIV-1 transmission. 25275130 6 71 theme trans-infected 1143:1156 arg1 lymphocytes 1166:1176 (iv) trans-infected primary lymphocytes 1138:1176 (iv) trans-infected primary lymphocytes 1138:1176 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 5 72 theme HIV-1 806:810 arg1 transmission 812:823 HIV-1 transmission 806:823 HIV-1 transmission 806:823 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 7 73 theme fresh 1332:1336 arg1 tissue 1355:1360 fresh human intestinal tissue 1332:1360 fresh human intestinal tissue 1332:1360 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 4 74 theme glycan 595:600 arg1 moieties 602:609 Env glycan moieties 591:609 Env glycan moieties 591:609 Also, the role of Env glycan moieties on HIV-1 transmission has not been systematically defined. 25275130 2 75 theme key 286:288 arg1 roles 290:294 key roles 286:294 key roles 286:294 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 1 76 theme Env 159:161 arg1 mass 173:176 the Env molecular mass 155:176 the Env molecular mass 155:176 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 6 77 theme more 912:915 arg1 oligomannose 917:928 more oligomannose 912:928 more oligomannose 912:928 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 10 78 theme IMPORTANCE 1676:1685 arg1 N-glycans 1697:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans 1676:1705 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 1 79 theme mass 173:176 arg1 mass 173:176 the Env molecular mass 155:176 the Env molecular mass 155:176 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 1 79 theme mass 173:176 arg1 % 150:150 approximately 50% 134:150 approximately 50% of the Env molecular mass 134:176 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 10 80 theme antibody 1854:1861 arg1 neutralization 1863:1876 antibody neutralization 1854:1876 antibody neutralization 1854:1876 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 2 81 theme Env 216:218 arg1 N-glycans 220:228 HIV-1 Env N-glycans 210:228 HIV-1 Env N-glycans 210:228 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 5 82 theme transmission 812:823 arg1 events 796:801 the major events 786:801 the major events of HIV-1 transmission 786:823 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 6 83 with glycans 947:953 arg1 oligomannose 917:928 more oligomannose 912:928 more oligomannose 912:928 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 84 link monocyte-derived 1092:1107 arg1 cells 1119:1123 monocyte-derived dendritic cells 1092:1123 monocyte-derived dendritic cells (MoDCs) 1092:1131 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 84 link monocyte-derived 1092:1107 arg1 MoDCs 1126:1130 MoDCs 1126:1130 MoDCs 1126:1130 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 85 theme complex 878:884 arg1 glycans 890:896 more complex Env glycans 873:896 more complex Env glycans 873:896 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 2 86 theme protein 241:247 arg1 backbone 249:256 the protein backbone 237:256 the protein backbone 237:256 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 13 87 theme Env 2184:2186 arg1 glycans 2188:2194 N-linked Env glycans 2175:2194 N-linked Env glycans 2175:2194 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 11 88 theme glycosylation 1935:1947 arg1 Studies 1918:1924 Studies 1918:1924 Studies of HIV-1 glycosylation 1918:1947 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 9 89 theme strategies 1632:1641 arg1 development 1609:1619 the development 1605:1619 the development of vaccine strategies to interdict HIV-1 transmission 1605:1673 Env glycosylation should be taken into consideration in the development of vaccine strategies to interdict HIV-1 transmission. 25275130 8 90 theme N-linked 1369:1376 arg1 glycans 1382:1388 N-linked Env glycans 1369:1388 N-linked Env glycans 1369:1388 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 5 91 theme glycan 702:707 arg1 content 709:715 modified Env glycan content 689:715 modified Env glycan content 689:715 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 3 92 theme glycans 564:570 arg1 types 555:559 the types 551:559 the types of glycans 551:570 Studies of HIV-1 glycosylation have focused mainly on the position of glycosylation, rather than the types of glycans. 25275130 5 93 theme moieties 774:781 arg1 effects 752:758 the effects 748:758 the effects of Env glycan moieties on the major events of HIV-1 transmission 748:823 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 7 94 theme complex 1238:1244 arg1 glycans 1246:1252 less complex glycans 1233:1252 less complex glycans 1233:1252 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 6 95 theme complex 939:945 arg1 glycans 947:953 less complex glycans 934:953 less complex glycans 934:953 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 8 96 theme discordant 1398:1407 arg1 effects 1409:1415 discordant effects 1398:1415 discordant effects on the major events of HIV-1 transmission 1398:1457 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 1 97 theme UNLABELLED 60:69 arg1 Env 99:101 Env 99:101 Env 99:101 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 1 97 theme UNLABELLED 60:69 arg1 protein 90:96 The HIV-1 envelope protein 71:96 UNLABELLED The HIV-1 envelope protein (Env) 60:102 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 12 98 theme glycan 2123:2128 arg1 moieties 2130:2137 Env glycan moieties 2119:2137 Env glycan moieties 2119:2137 In the study described in this report, we investigated systematically the role of Env glycan moieties on HIV-1 transmission. 25275130 5 99 theme modified 689:696 arg1 content 709:715 modified Env glycan content 689:715 modified Env glycan content 689:715 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 1 100 theme HIV-1 75:79 arg1 Env 99:101 Env 99:101 Env 99:101 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 1 100 theme HIV-1 75:79 arg1 protein 90:96 The HIV-1 envelope protein 71:96 UNLABELLED The HIV-1 envelope protein (Env) 60:102 UNLABELLED The HIV-1 envelope protein (Env) is heavily glycosylated, with approximately 50% of the Env molecular mass being contributed by N-glycans. 25275130 8 101 theme major 1424:1428 arg1 events 1430:1435 the major events 1420:1435 the major events of HIV-1 transmission 1420:1457 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 6 102 dep transcytosed 976:987 arg1 i 973:973 i 973:973 i 973:973 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 0 103 theme envelope 6:13 arg1 moieties 22:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties 0:29 HIV-1 envelope glycan moieties modulate HIV-1 transmission. 25275130 6 104 theme epithelial 999:1008 arg1 monolayer 1015:1023 an epithelial cell monolayer 996:1023 an epithelial cell monolayer 996:1023 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 5 105 theme Env 763:765 arg1 moieties 774:781 Env glycan moieties 763:781 Env glycan moieties 763:781 Using viruses with modified Env glycan content and heterogeneity, we examined the effects of Env glycan moieties on the major events of HIV-1 transmission. 25275130 7 106 from infectivity 1273:1283 arg1 MDMs 1301:1304 MDMs 1301:1304 MDMs 1301:1304 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 7 106 from infectivity 1273:1283 arg1 lymphocytes 1315:1325 primary lymphocytes 1307:1325 primary lymphocytes 1307:1325 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 7 106 from infectivity 1273:1283 arg1 cells 1294:1298 TZMbl cells 1288:1298 TZMbl cells 1288:1298 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 7 106 from infectivity 1273:1283 arg1 tissue 1355:1360 fresh human intestinal tissue 1332:1360 fresh human intestinal tissue 1332:1360 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 8 107 link N-linked 1369:1376 arg1 glycans 1382:1388 N-linked Env glycans 1369:1388 N-linked Env glycans 1369:1388 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 8 108 theme transmission 1446:1457 arg1 events 1430:1435 the major events 1420:1435 the major events of HIV-1 transmission 1420:1457 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 10 109 theme Env 1769:1771 arg1 structure 1773:1781 Env structure 1769:1781 Env structure 1769:1781 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 0 110 theme HIV-1 40:44 arg1 transmission 46:57 HIV-1 transmission 40:57 HIV-1 transmission 40:57 HIV-1 envelope glycan moieties modulate HIV-1 transmission. 25275130 4 111 theme Env 591:593 arg1 moieties 602:609 Env glycan moieties 591:609 Env glycan moieties 591:609 Also, the role of Env glycan moieties on HIV-1 transmission has not been systematically defined. 25275130 6 112 theme monocyte-derived 1092:1107 arg1 cells 1119:1123 monocyte-derived dendritic cells 1092:1123 monocyte-derived dendritic cells (MoDCs) 1092:1131 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 6 112 theme monocyte-derived 1092:1107 arg1 MoDCs 1126:1130 MoDCs 1126:1130 MoDCs 1126:1130 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 13 113 from effects 2215:2221 arg1 events 2236:2241 the major events 2226:2241 the major events of HIV-1 transmission 2226:2263 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 14 114 theme therapeutic 2431:2441 arg1 vaccines 2459:2466 therapeutic or prophylactic vaccines 2431:2466 therapeutic or prophylactic vaccines against HIV-1 2431:2480 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 7 115 with viruses 1198:1204 arg1 glycans 1246:1252 less complex glycans 1233:1252 less complex glycans 1233:1252 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 7 115 with viruses 1198:1204 arg1 oligomannose 1216:1227 more oligomannose 1211:1227 more oligomannose 1211:1227 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 8 116 theme oligosaccharide 1472:1486 arg1 structures 1488:1497 mature oligosaccharide structures 1465:1497 mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection 1465:1546 Thus, N-linked Env glycans display discordant effects on the major events of HIV-1 transmission, with mature oligosaccharide structures on Env playing a crucial role in HIV-1 infection. 25275130 13 117 theme HIV-1 2246:2250 arg1 transmission 2252:2263 HIV-1 transmission 2246:2263 HIV-1 transmission 2246:2263 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 7 118 theme TZMbl 1288:1292 arg1 cells 1294:1298 TZMbl cells 1288:1298 TZMbl cells 1288:1298 However, viruses with more oligomannose and less complex glycans displayed impaired infectivity in TZMbl cells, MDMs, primary lymphocytes, and fresh human intestinal tissue. 25275130 13 119 link N-linked 2175:2182 arg1 glycans 2188:2194 N-linked Env glycans 2175:2194 N-linked Env glycans 2175:2194 We show that N-linked Env glycans display discordant effects on the major events of HIV-1 transmission. 25275130 6 120 dep attached 1031:1038 arg1 ii 1027:1028 ii 1027:1028 ii 1027:1028 Compared to viruses with less oligomannose and more complex Env glycans, viruses with more oligomannose and less complex glycans more efficiently (i) transcytosed across an epithelial cell monolayer, (ii) attached to monocyte-derived macrophages (MDMs), (iii) bound monocyte-derived dendritic cells (MoDCs), and (iv) trans-infected primary lymphocytes via MoDCs. 25275130 14 121 theme HIV-1 2318:2322 arg1 transmission 2324:2335 HIV-1 transmission 2318:2335 HIV-1 transmission 2318:2335 These data indicate that Env glycan moieties impact HIV-1 transmission and that modulation of Env glycan moieties offers a potential strategy for the development of therapeutic or prophylactic vaccines against HIV-1. 25275130 10 122 theme Env 1823:1825 arg1 antigenicity 1827:1838 Env antigenicity 1823:1838 Env antigenicity 1823:1838 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 10 123 theme protein 1718:1724 arg1 backbone 1726:1733 the protein backbone 1714:1733 the protein backbone 1714:1733 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 2 124 theme surface 326:332 arg1 exposure 334:341 surface exposure 326:341 surface exposure 326:341 HIV-1 Env N-glycans shield the protein backbone and have been shown to play key roles in determining Env structure, surface exposure, and, consequently, antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 25275130 10 125 theme receptor 1900:1907 arg1 binding 1909:1915 receptor binding 1900:1915 receptor binding 1900:1915 IMPORTANCE HIV-1 Env N-glycans shield the protein backbone and play key roles in determining Env structure and surface exposure, thereby impacting Env antigenicity, infectivity, antibody neutralization, and carbohydrate and receptor binding. 27718394 9 0 theme protein 1685:1691 arg1 experiment 1704:1713 a protein array-type experiment 1683:1713 a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1683:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 9 1 theme experiment 1704:1713 arg1 example 1672:1678 the first example 1662:1678 the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1662:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 9 1 theme experiment 1704:1713 arg1 this 1654:1657 this 1654:1657 this 1654:1657 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 8 2 theme developed 1407:1415 arg1 method 1417:1422 The developed method 1403:1422 The developed method 1403:1422 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 2 3 theme entire 311:316 arg1 population 326:335 the entire protein population 307:335 the entire protein population of a sample 307:347 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 9 4 from fluid 1890:1894 arg1 analysis 1844:1851 fast and efficient glycan analysis 1818:1851 fast and efficient glycan analysis of individual proteins in biological fluid 1818:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 9 5 theme immunoaffinity 1739:1752 arg1 purification 1754:1765 bead-based immunoaffinity purification 1728:1765 bead-based immunoaffinity purification 1728:1765 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 2 6 from variations 376:385 arg1 concentrations 394:407 the concentrations 390:407 the concentrations of certain proteins 390:427 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 2 7 gly glycosylation 290:302 arg1 population 326:335 the entire protein population 307:335 the entire protein population of a sample 307:347 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 6 8 theme targeted 1281:1288 arg1 protein 1290:1296 the targeted protein 1277:1296 the targeted protein 1277:1296 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 8 9 gly glycosylation 1510:1522 arg1 alpha-1-antitrypsin 1559:1577 alpha-1-antitrypsin 1559:1577 alpha-1-antitrypsin 1559:1577 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 8 9 gly glycosylation 1510:1522 arg1 proteins 1549:1556 two different proteins 1535:1556 two different proteins 1535:1556 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 8 9 gly glycosylation 1510:1522 arg1 transferrin 1583:1593 transferrin 1583:1593 transferrin 1583:1593 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 6 10 theme VHHs 1198:1201 arg1 advantage 1181:1189 A major advantage 1173:1189 A major advantage of the VHHs 1173:1201 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 5 11 theme immunoaffinity 1118:1131 arg1 binders 1133:1139 genetically engineered immunoaffinity binders 1095:1139 genetically engineered immunoaffinity binders 1095:1139 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 2 12 theme population 326:335 arg1 glycosylation 290:302 the glycosylation 286:302 the glycosylation of the entire protein population of a sample 286:347 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 1 13 theme chemical 225:232 arg1 properties 253:262 both the chemical and the biological properties 216:262 properties 253:262 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 3 14 theme different 643:651 arg1 types 653:657 different types 643:657 different types of cancer, as well as inflammatory and neurodegenerative diseases 643:723 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 9 15 theme spectrometry 1792:1803 arg1 analysis 1805:1812 mass spectrometry analysis 1787:1812 mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1787:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 1 16 dep properties 253:262 arg1 both 216:219 both 216:219 both 216:219 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 6 17 theme glycans 1264:1270 arg1 release 1253:1259 enzymatic release 1243:1259 enzymatic release of glycans from the targeted protein 1243:1296 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 4 18 theme rapid 754:758 arg1 method 774:779 a rapid and efficient method 752:779 a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 752:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 3 19 theme disease 589:595 arg1 states 597:602 disease states 589:602 disease states 589:602 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 4 20 from changes 843:849 arg1 degree 858:863 the degree 854:863 the degree of fucosylation 854:879 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 4 21 theme modified 1001:1008 arg1 degree 1010:1015 a modified degree 999:1015 a modified degree of branching 999:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 0 22 theme biological 66:75 arg1 samples 77:83 biological samples 66:83 biological samples 66:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 9 23 theme glycan 1837:1842 arg1 analysis 1844:1851 fast and efficient glycan analysis 1818:1851 fast and efficient glycan analysis of individual proteins in biological fluid 1818:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 8 24 theme proteins 1549:1556 arg1 pattern 1524:1530 the glycosylation pattern 1506:1530 the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid 1506:1633 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 9 25 theme fast 1818:1821 arg1 analysis 1844:1851 fast and efficient glycan analysis 1818:1851 fast and efficient glycan analysis of individual proteins in biological fluid 1818:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 1 26 theme post-translational 170:187 arg1 modifications 189:201 the most common and important post-translational modifications 140:201 the most common and important post-translational modifications 140:201 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 4 27 theme bisecting 964:972 arg1 N-acetylglucosamines 974:993 bisecting N-acetylglucosamines 964:993 bisecting N-acetylglucosamines 964:993 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 5 28 theme VHH 1149:1151 arg1 fragments 1162:1170 VHH antibody fragments 1149:1170 VHH antibody fragments 1149:1170 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 9 29 theme proteins 1867:1874 arg1 analysis 1844:1851 fast and efficient glycan analysis 1818:1851 fast and efficient glycan analysis of individual proteins in biological fluid 1818:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 4 30 theme individual 811:820 arg1 proteins 822:829 individual proteins 811:829 individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 811:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 8 31 from pattern 1524:1530 arg1 serum 1605:1609 human serum 1599:1609 human serum 1599:1609 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 8 31 from pattern 1524:1530 arg1 fluid 1629:1633 cerebrospinal fluid 1615:1633 cerebrospinal fluid 1615:1633 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 0 32 gly nonglycosylated 91:105 arg1 binders 107:113 nonglycosylated binders 91:113 nonglycosylated binders 91:113 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 9 33 theme biological 1879:1888 arg1 fluid 1890:1894 biological fluid 1879:1894 biological fluid 1879:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 4 34 theme efficient 764:772 arg1 method 774:779 a rapid and efficient method 752:779 a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 752:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 2 35 theme certain 412:418 arg1 proteins 420:427 certain proteins 412:427 certain proteins 412:427 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 0 36 theme Particle-based 0:13 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 0 37 theme glycan 24:29 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 9 38 from proteins 1867:1874 arg1 fluid 1890:1894 biological fluid 1879:1894 biological fluid 1879:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 8 39 theme cerebrospinal 1615:1627 arg1 fluid 1629:1633 cerebrospinal fluid 1615:1633 cerebrospinal fluid 1615:1633 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 3 40 theme inflammatory 681:692 arg1 diseases 716:723 inflammatory and neurodegenerative diseases 681:723 diseases 716:723 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 3 41 from alterations 491:501 arg1 pattern 524:530 the glycosylation pattern 506:530 the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, 506:603 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 0 42 theme proteins 52:59 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 9 43 theme bead-based 1728:1737 arg1 purification 1754:1765 bead-based immunoaffinity purification 1728:1765 bead-based immunoaffinity purification 1728:1765 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 3 44 theme glycosylation 510:522 arg1 pattern 524:530 the glycosylation pattern 506:530 the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, 506:603 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 1 45 theme biological 242:251 arg1 properties 253:262 both the chemical and the biological properties 216:262 properties 253:262 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 3 46 theme neurodegenerative 698:714 arg1 diseases 716:723 inflammatory and neurodegenerative diseases 681:723 diseases 716:723 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 5 47 theme magnetic 1051:1058 arg1 beads 1060:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 2 48 from changes 452:458 arg1 glycosylation 463:475 glycosylation 463:475 glycosylation 463:475 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 4 49 attach present 744:750 arg1 paper 734:738 this paper 729:738 this paper 729:738 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 4 49 attach present 744:750 arg2 we 741:742 we 741:742 we 741:742 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 3 50 theme proteins 546:553 arg1 pattern 524:530 the glycosylation pattern 506:530 the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, 506:603 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 6 51 from protein 1290:1296 arg1 release 1253:1259 enzymatic release 1243:1259 enzymatic release of glycans from the targeted protein 1243:1296 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 9 52 theme array-type 1693:1702 arg1 experiment 1704:1713 a protein array-type experiment 1683:1713 a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1683:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 2 53 theme protein 318:324 arg1 population 326:335 the entire protein population 307:335 the entire protein population of a sample 307:347 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 4 54 theme core 909:912 arg1 structure 914:922 the core structure 905:922 the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 905:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 0 55 link N-linked 15:22 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 1 56 theme modifications 189:201 arg1 one 133:135 one 133:135 one 133:135 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 1 56 theme modifications 189:201 arg1 modifications 189:201 the most common and important post-translational modifications 140:201 the most common and important post-translational modifications 140:201 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 4 57 theme fucosylation 868:879 arg1 degree 858:863 the degree 854:863 the degree of fucosylation 854:879 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 2 58 theme sample 342:347 arg1 population 326:335 the entire protein population 307:335 the entire protein population of a sample 307:347 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 4 59 theme other 884:888 arg1 alterations 890:900 other alterations 884:900 other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 884:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 6 60 gly nonglycosylated 1220:1234 arg1 they 1211:1214 they 1211:1214 they 1211:1214 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 3 61 theme individual 535:544 arg1 proteins 546:553 individual proteins 535:553 individual proteins 535:553 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 9 62 theme mass 1787:1790 arg1 analysis 1805:1812 mass spectrometry analysis 1787:1812 mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1787:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 8 63 theme glycosylation 1510:1522 arg1 pattern 1524:1530 the glycosylation pattern 1506:1530 the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid 1506:1633 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 3 64 theme cancer 662:667 arg1 types 653:657 different types 643:657 different types of cancer, as well as inflammatory and neurodegenerative diseases 643:723 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 4 65 theme glycans 931:937 arg1 structure 914:922 the core structure 905:922 the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 905:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 8 66 theme different 1539:1547 arg1 alpha-1-antitrypsin 1559:1577 alpha-1-antitrypsin 1559:1577 alpha-1-antitrypsin 1559:1577 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 8 66 theme different 1539:1547 arg1 proteins 1549:1556 two different proteins 1535:1556 two different proteins 1535:1556 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 8 66 theme different 1539:1547 arg1 transferrin 1583:1593 transferrin 1583:1593 transferrin 1583:1593 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 6 67 theme enzymatic 1243:1251 arg1 release 1253:1259 enzymatic release 1243:1259 enzymatic release of glycans from the targeted protein 1243:1296 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 4 68 gly glycosylation 785:797 arg1 proteins 822:829 individual proteins 811:829 individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 811:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 5 69 theme engineered 1107:1116 arg1 binders 1133:1139 genetically engineered immunoaffinity binders 1095:1139 genetically engineered immunoaffinity binders 1095:1139 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 1 70 theme common 149:154 arg1 modifications 189:201 the most common and important post-translational modifications 140:201 the most common and important post-translational modifications 140:201 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 9 71 theme efficient 1827:1835 arg1 analysis 1844:1851 fast and efficient glycan analysis 1818:1851 fast and efficient glycan analysis of individual proteins in biological fluid 1818:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 0 72 from analysis 31:38 arg1 samples 77:83 biological samples 66:83 biological samples 66:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 3 73 from variability 574:584 arg1 states 597:602 disease states 589:602 disease states 589:602 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 0 74 theme nonglycosylated 91:105 arg1 binders 107:113 nonglycosylated binders 91:113 nonglycosylated binders 91:113 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 1 75 theme important 160:168 arg1 modifications 189:201 the most common and important post-translational modifications 140:201 the most common and important post-translational modifications 140:201 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 27718394 4 76 from alterations 890:900 arg1 degree 858:863 the degree 854:863 the degree of fucosylation 854:879 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 4 77 theme degree 1010:1015 arg1 presence 952:959 the presence 948:959 the presence of bisecting N-acetylglucosamines and a modified degree of branching 948:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 9 78 theme individual 1856:1865 arg1 proteins 1867:1874 individual proteins 1856:1874 individual proteins in biological fluid 1856:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 4 79 theme glycosylation 785:797 arg1 analysis 799:806 glycosylation analysis 785:806 glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 785:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 5 80 theme antibody 1153:1160 arg1 fragments 1162:1170 VHH antibody fragments 1149:1170 VHH antibody fragments 1149:1170 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 4 81 theme N-acetylglucosamines 974:993 arg1 presence 952:959 the presence 948:959 the presence of bisecting N-acetylglucosamines and a modified degree of branching 948:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 8 82 theme human 1599:1603 arg1 serum 1605:1609 human serum 1599:1609 human serum 1599:1609 The developed method was evaluated concerning its specificity, and thereafter implemented for studying the glycosylation pattern of two different proteins, alpha-1-antitrypsin and transferrin, in human serum and cerebrospinal fluid. 27718394 5 83 with combination 1078:1088 arg1 binders 1133:1139 genetically engineered immunoaffinity binders 1095:1139 genetically engineered immunoaffinity binders 1095:1139 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 2 84 theme proteins 420:427 arg1 concentrations 394:407 the concentrations 390:407 the concentrations of certain proteins 390:427 Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. 27718394 0 85 theme N-linked 15:22 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 3 86 gly glycosylation 510:522 arg1 proteins 546:553 individual proteins 535:553 individual proteins 535:553 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 0 87 theme selected 43:50 arg1 proteins 52:59 selected proteins 43:59 selected proteins from biological samples 43:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 4 88 theme branching 1020:1028 arg1 N-acetylglucosamines 974:993 bisecting N-acetylglucosamines 964:993 bisecting N-acetylglucosamines 964:993 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 4 88 theme branching 1020:1028 arg1 degree 1010:1015 a modified degree 999:1015 a modified degree of branching 999:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 9 89 with combination 1770:1780 arg1 analysis 1805:1812 mass spectrometry analysis 1787:1812 mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1787:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 3 90 theme diseases 716:723 arg1 types 653:657 different types 643:657 different types of cancer, as well as inflammatory and neurodegenerative diseases 643:723 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 0 91 from samples 77:83 arg1 analysis 31:38 Particle-based N-linked glycan analysis 0:38 Particle-based N-linked glycan analysis of selected proteins from biological samples 0:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 0 91 from samples 77:83 arg1 proteins 52:59 selected proteins 43:59 selected proteins from biological samples 43:83 Particle-based N-linked glycan analysis of selected proteins from biological samples using nonglycosylated binders. 27718394 9 92 from analysis 1844:1851 arg1 fluid 1890:1894 biological fluid 1879:1894 biological fluid 1879:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 9 93 theme first 1666:1670 arg1 example 1672:1678 the first example 1662:1678 the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid 1662:1894 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 9 93 theme first 1666:1670 arg1 this 1654:1657 this 1654:1657 this 1654:1657 To our knowledge, this is the first example of a protein array-type experiment that employs bead-based immunoaffinity purification in combination with mass spectrometry analysis for fast and efficient glycan analysis of individual proteins in biological fluid. 27718394 6 94 theme major 1175:1179 arg1 advantage 1181:1189 A major advantage 1173:1189 A major advantage of the VHHs 1173:1201 A major advantage of the VHHs is that they are nonglycosylated; thus, enzymatic release of glycans from the targeted protein can be performed directly on the beads. 27718394 4 95 theme proteins 822:829 arg1 analysis 799:806 glycosylation analysis 785:806 glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching 785:1028 In this paper, we present a rapid and efficient method for glycosylation analysis of individual proteins focusing on changes in the degree of fucosylation or other alterations to the core structure of the glycans, such as the presence of bisecting N-acetylglucosamines and a modified degree of branching. 27718394 3 96 theme larger 567:572 arg1 variability 574:584 larger variability 567:584 larger variability in disease states 567:602 Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases. 27718394 5 97 theme Streptavidin-coated 1031:1049 arg1 beads 1060:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 5 98 used used 1070:1073 arg2 beads 1060:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads 1031:1064 Streptavidin-coated magnetic beads are used in combination with genetically engineered immunoaffinity binders, called VHH antibody fragments. 27718394 1 99 theme proteins 267:274 arg1 properties 253:262 both the chemical and the biological properties 216:262 properties 253:262 Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. 25118277 7 0 theme O-glycophenotype 1573:1588 arg1 evaluation 1548:1557 the first polyomic and side-by-side evaluation 1512:1557 the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1512:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 4 1 theme glycosylation 968:980 arg1 functions 946:954 the biological functions 931:954 the biological functions of aberrant glycosylation in cancer 931:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 3 2 theme O-glycosylation 759:773 arg1 initiation 745:754 initiation 745:754 initiation of O-glycosylation from Golgi to endoplasmic reticulum 745:809 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 2 3 theme structures 379:388 arg1 Expression 342:351 Expression 342:351 Expression of the truncated O-glycan structures Tn and sialyl-Tn 342:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 1 4 theme epithelial 261:270 arg1 lesions 285:291 early epithelial premalignant lesions 255:291 early epithelial premalignant lesions that precede the development of adenocarcinomas 255:339 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 6 5 theme molecular 1282:1290 arg1 chaperone 1292:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 5 theme molecular 1282:1290 arg1 chaperone 1309:1317 a key chaperone 1303:1317 a key chaperone for O-glycan elongation 1303:1341 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 6 theme prevalent 1356:1364 arg1 hypermethylation 1237:1252 hypermethylation 1237:1252 hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, 1237:1342 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 6 theme prevalent 1356:1364 arg1 cause 1366:1370 the most prevalent cause 1347:1370 the most prevalent cause 1347:1370 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 4 7 theme biological 935:944 arg1 functions 946:954 the biological functions 931:954 the biological functions of aberrant glycosylation in cancer 931:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 3 8 theme endoplasmic 789:799 arg1 reticulum 801:809 endoplasmic reticulum 789:809 endoplasmic reticulum 789:809 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 1 9 theme cancer 200:205 arg1 cells 207:211 virtually all epithelial cancer cells 175:211 virtually all epithelial cancer cells 175:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 5 10 theme exome 1031:1035 arg1 sequencing 1037:1046 exome sequencing 1031:1046 exome sequencing of most glycosyltransferases 1031:1075 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 5 10 theme exome 1031:1035 arg1 cause 1175:1179 a cause 1173:1179 a cause of expression of truncated O-glycans 1173:1216 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 3 11 theme O-glycans 553:561 arg1 accumulation 527:538 accumulation 527:538 accumulation of truncated O-glycans 527:561 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 9 12 theme truncated 1820:1828 arg1 O-glycans 1830:1838 cancer-specific truncated O-glycans 1804:1838 cancer-specific truncated O-glycans with immunotherapeutic measures 1804:1870 The study provides support for targeting cancer-specific truncated O-glycans with immunotherapeutic measures. 25118277 1 13 theme high 225:228 arg1 frequency 230:238 a very high frequency 218:238 a very high frequency 218:238 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 14 theme homogenous 1449:1458 arg1 O-glycans 1470:1478 homogenous truncated O-glycans 1449:1478 homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1449:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 6 15 theme key 1305:1307 arg1 chaperone 1292:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 15 theme key 1305:1307 arg1 chaperone 1309:1317 a key chaperone 1303:1317 a key chaperone for O-glycan elongation 1303:1341 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 4 16 theme Truncated 812:820 arg1 O-glycans 822:830 Truncated O-glycans 812:830 Truncated O-glycans 812:830 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 3 17 theme biosynthetic 492:503 arg1 mechanisms 505:514 The genetic and biosynthetic mechanisms 476:514 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans 476:561 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 5 18 theme O-glycans 1208:1216 arg1 expression 1184:1193 expression 1184:1193 expression of truncated O-glycans 1184:1216 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 3 19 theme O-glycans 676:684 arg1 elongation 662:671 elongation 662:671 elongation of O-glycans 662:684 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 4 20 from glycosylation 968:980 arg1 cancer 985:990 cancer 985:990 cancer 985:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 1 21 theme all 185:187 arg1 cells 207:211 virtually all epithelial cancer cells 175:211 virtually all epithelial cancer cells 175:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 6 22 theme β3-Gal-T-specific 1264:1280 arg1 chaperone 1292:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 22 theme β3-Gal-T-specific 1264:1280 arg1 chaperone 1309:1317 a key chaperone 1303:1317 a key chaperone for O-glycan elongation 1303:1341 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 5 23 theme glycosyltransferases 1056:1075 arg1 sequencing 1037:1046 exome sequencing 1031:1046 exome sequencing of most glycosyltransferases 1031:1075 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 5 23 theme glycosyltransferases 1056:1075 arg1 cause 1175:1179 a cause 1173:1179 a cause of expression of truncated O-glycans 1173:1216 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 7 24 theme cell 1419:1422 arg1 systems 1424:1430 isogenic cell systems 1410:1430 isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1410:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 7 25 used used 1381:1384 arg2 We 1373:1374 We 1373:1374 We 1373:1374 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 0 26 theme Immature 0:7 arg1 O-glycophenotype 19:34 Immature truncated O-glycophenotype 0:34 Immature truncated O-glycophenotype of cancer 0:44 Immature truncated O-glycophenotype of cancer directly induces oncogenic features. 25118277 1 27 theme immature 106:113 arg1 O-glycans 125:133 immature truncated O-glycans 106:133 immature truncated O-glycans 106:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 2 28 theme overall 454:460 arg1 survival 466:473 overall low survival 454:473 overall low survival 454:473 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 1 29 theme O-glycans 125:133 arg1 expression 92:101 Aberrant expression 83:101 Aberrant expression of immature truncated O-glycans 83:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 29 theme O-glycans 125:133 arg1 feature 155:161 a characteristic feature 138:161 a characteristic feature observed on virtually all epithelial cancer cells 138:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 3 30 theme genetic 480:486 arg1 mechanisms 505:514 The genetic and biosynthetic mechanisms 476:514 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans 476:561 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 0 31 theme cancer 39:44 arg1 O-glycophenotype 19:34 Immature truncated O-glycophenotype 0:34 Immature truncated O-glycophenotype of cancer 0:44 Immature truncated O-glycophenotype of cancer directly induces oncogenic features. 25118277 5 32 theme metastatic 1110:1119 arg1 cancers 1132:1138 primary and metastatic pancreatic cancers 1098:1138 primary and metastatic pancreatic cancers 1098:1138 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 2 33 theme O-glycan 370:377 arg1 Tn 390:391 Tn 390:391 Tn 390:391 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 33 theme O-glycan 370:377 arg1 structures 379:388 the truncated O-glycan structures Tn and sialyl-Tn 356:405 the truncated O-glycan structures Tn and sialyl-Tn 356:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 33 theme O-glycan 370:377 arg1 sialyl-Tn 397:405 sialyl-Tn 397:405 sialyl-Tn 397:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 4 34 from cancer 985:990 arg1 functions 946:954 the biological functions 931:954 the biological functions of aberrant glycosylation in cancer 931:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 7 35 theme organotypic 1596:1606 arg1 model 1615:1619 an organotypic tissue model 1593:1619 an organotypic tissue model 1593:1619 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 5 36 theme cancers 1132:1138 arg1 series 1088:1093 a large series 1080:1093 a large series of primary and metastatic pancreatic cancers to rule out somatic mutations 1080:1168 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 4 37 theme cancer-cell 880:890 arg1 invasiveness 892:903 cancer-cell invasiveness 880:903 cancer-cell invasiveness 880:903 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 8 38 theme O-glycans 1688:1696 arg1 truncation 1674:1683 truncation 1674:1683 truncation of O-glycans 1674:1696 The results strongly suggest that truncation of O-glycans directly induces oncogenic features of cell growth and invasion. 25118277 4 39 gly glycosylation 968:980 arg1 cancer 985:990 cancer 985:990 cancer 985:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 7 40 theme cancer 1566:1571 arg1 O-glycophenotype 1573:1588 the cancer O-glycophenotype 1562:1588 the cancer O-glycophenotype 1562:1588 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 5 41 theme primary 1098:1104 arg1 cancers 1132:1138 primary and metastatic pancreatic cancers 1098:1138 primary and metastatic pancreatic cancers 1098:1138 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 3 42 theme truncated 543:551 arg1 O-glycans 553:561 truncated O-glycans 543:561 truncated O-glycans 543:561 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 1 43 theme early 255:259 arg1 lesions 285:291 early epithelial premalignant lesions 255:291 early epithelial premalignant lesions that precede the development of adenocarcinomas 255:339 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 4 44 theme aberrant 959:966 arg1 glycosylation 968:980 aberrant glycosylation 959:980 aberrant glycosylation in cancer 959:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 1 45 theme premalignant 272:283 arg1 lesions 285:291 early epithelial premalignant lesions 255:291 early epithelial premalignant lesions that precede the development of adenocarcinomas 255:339 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 46 theme side-by-side 1535:1546 arg1 evaluation 1548:1557 the first polyomic and side-by-side evaluation 1512:1557 the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1512:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 7 47 theme first 1516:1520 arg1 evaluation 1548:1557 the first polyomic and side-by-side evaluation 1512:1557 the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1512:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 8 48 theme cell 1737:1740 arg1 features 1725:1732 oncogenic features 1715:1732 oncogenic features of cell growth and invasion 1715:1760 The results strongly suggest that truncation of O-glycans directly induces oncogenic features of cell growth and invasion. 25118277 1 49 theme epithelial 189:198 arg1 cells 207:211 virtually all epithelial cancer cells 175:211 virtually all epithelial cancer cells 175:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 50 theme polyomic 1522:1529 arg1 evaluation 1548:1557 the first polyomic and side-by-side evaluation 1512:1557 the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1512:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 6 51 theme core 1257:1260 arg1 chaperone 1292:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 core 1 β3-Gal-T-specific molecular chaperone 1257:1300 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 51 theme core 1257:1260 arg1 chaperone 1309:1317 a key chaperone 1303:1317 a key chaperone for O-glycan elongation 1303:1341 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 4 52 from functions 946:954 arg1 cancer 985:990 cancer 985:990 cancer 985:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 4 53 theme functions 946:954 arg1 understanding 914:926 our understanding 910:926 our understanding of the biological functions of aberrant glycosylation in cancer 910:990 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 1 54 located observed 243:250 arg2 frequency 230:238 a very high frequency 218:238 a very high frequency 218:238 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 54 located observed 243:250 arg1 lesions 285:291 early epithelial premalignant lesions 255:291 early epithelial premalignant lesions that precede the development of adenocarcinomas 255:339 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 55 from evaluation 1548:1557 arg1 model 1615:1619 an organotypic tissue model 1593:1619 an organotypic tissue model 1593:1619 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 7 55 from evaluation 1548:1557 arg1 xenografts 1628:1637 xenografts 1628:1637 xenografts 1628:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 9 56 theme cancer-specific 1804:1818 arg1 O-glycans 1830:1838 cancer-specific truncated O-glycans 1804:1838 cancer-specific truncated O-glycans with immunotherapeutic measures 1804:1870 The study provides support for targeting cancer-specific truncated O-glycans with immunotherapeutic measures. 25118277 6 57 theme O-glycan 1323:1330 arg1 elongation 1332:1341 O-glycan elongation 1323:1341 O-glycan elongation 1323:1341 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 3 58 from Golgi 780:784 arg1 initiation 745:754 initiation 745:754 initiation of O-glycosylation from Golgi to endoplasmic reticulum 745:809 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 3 59 theme glycosyltransferases 629:648 arg1 relocation 698:707 relocation 698:707 relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum 698:809 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 3 59 theme glycosyltransferases 629:648 arg1 dysregulation 612:624 dysregulation 612:624 dysregulation 612:624 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 3 59 theme glycosyltransferases 629:648 arg1 mutation 600:607 mutation 600:607 mutation 600:607 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 9 60 theme immunotherapeutic 1845:1861 arg1 measures 1863:1870 immunotherapeutic measures 1845:1870 immunotherapeutic measures 1845:1870 The study provides support for targeting cancer-specific truncated O-glycans with immunotherapeutic measures. 25118277 7 61 theme truncated 1460:1468 arg1 O-glycans 1470:1478 homogenous truncated O-glycans 1449:1478 homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1449:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 1 62 theme characteristic 140:153 arg1 feature 155:161 a characteristic feature 138:161 a characteristic feature observed on virtually all epithelial cancer cells 138:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 62 theme characteristic 140:153 arg1 expression 92:101 Aberrant expression 83:101 Aberrant expression of immature truncated O-glycans 83:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 5 63 theme somatic 1152:1158 arg1 mutations 1160:1168 somatic mutations 1152:1168 somatic mutations 1152:1168 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 6 64 theme chaperone 1292:1300 arg1 hypermethylation 1237:1252 hypermethylation 1237:1252 hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, 1237:1342 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 6 64 theme chaperone 1292:1300 arg1 cause 1366:1370 the most prevalent cause 1347:1370 the most prevalent cause 1347:1370 Instead, we found hypermethylation of core 1 β3-Gal-T-specific molecular chaperone, a key chaperone for O-glycan elongation, as the most prevalent cause. 25118277 8 65 dep cell 1737:1740 arg1 invasion 1753:1760 invasion 1753:1760 invasion 1753:1760 The results strongly suggest that truncation of O-glycans directly induces oncogenic features of cell growth and invasion. 25118277 8 65 dep cell 1737:1740 arg1 growth 1742:1747 growth 1742:1747 growth 1742:1747 The results strongly suggest that truncation of O-glycans directly induces oncogenic features of cell growth and invasion. 25118277 5 66 theme most 1051:1054 arg1 glycosyltransferases 1056:1075 most glycosyltransferases 1051:1075 most glycosyltransferases 1051:1075 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 2 67 dep structures 379:388 arg1 Tn 390:391 Tn 390:391 Tn 390:391 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 67 dep structures 379:388 arg1 structures 379:388 the truncated O-glycan structures Tn and sialyl-Tn 356:405 the truncated O-glycan structures Tn and sialyl-Tn 356:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 67 dep structures 379:388 arg1 sialyl-Tn 397:405 sialyl-Tn 397:405 sialyl-Tn 397:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 1 68 located observed 163:170 arg1 cells 207:211 virtually all epithelial cancer cells 175:211 virtually all epithelial cancer cells 175:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 68 located observed 163:170 arg2 expression 92:101 Aberrant expression 83:101 Aberrant expression of immature truncated O-glycans 83:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 68 located observed 163:170 arg2 feature 155:161 a characteristic feature 138:161 a characteristic feature observed on virtually all epithelial cancer cells 138:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 69 theme Aberrant 83:90 arg1 expression 92:101 Aberrant expression 83:101 Aberrant expression of immature truncated O-glycans 83:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 1 69 theme Aberrant 83:90 arg1 feature 155:161 a characteristic feature 138:161 a characteristic feature observed on virtually all epithelial cancer cells 138:211 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 70 theme isogenic 1410:1417 arg1 systems 1424:1430 isogenic cell systems 1410:1430 isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts 1410:1637 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 4 71 theme functional 859:868 arg1 roles 870:874 functional roles 859:874 functional roles 859:874 Truncated O-glycans have been proposed to play functional roles for cancer-cell invasiveness, but our understanding of the biological functions of aberrant glycosylation in cancer is still highly limited. 25118277 0 72 theme truncated 9:17 arg1 O-glycophenotype 19:34 Immature truncated O-glycophenotype 0:34 Immature truncated O-glycophenotype of cancer 0:44 Immature truncated O-glycophenotype of cancer directly induces oncogenic features. 25118277 5 73 theme truncated 1198:1206 arg1 O-glycans 1208:1216 truncated O-glycans 1198:1216 truncated O-glycans 1198:1216 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 5 74 theme expression 1184:1193 arg1 sequencing 1037:1046 exome sequencing 1031:1046 exome sequencing of most glycosyltransferases 1031:1075 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 5 74 theme expression 1184:1193 arg1 cause 1175:1179 a cause 1173:1179 a cause of expression of truncated O-glycans 1173:1216 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 5 75 used used 1026:1029 arg2 we 1023:1024 we 1023:1024 we 1023:1024 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 1 76 theme truncated 115:123 arg1 O-glycans 125:133 immature truncated O-glycans 106:133 immature truncated O-glycans 106:133 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 7 77 theme gene 1386:1389 arg1 editing 1391:1397 gene editing 1386:1397 gene editing 1386:1397 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 2 78 theme low 462:464 arg1 survival 466:473 overall low survival 454:473 overall low survival 454:473 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 9 79 with O-glycans 1830:1838 arg1 measures 1863:1870 immunotherapeutic measures 1845:1870 immunotherapeutic measures 1845:1870 The study provides support for targeting cancer-specific truncated O-glycans with immunotherapeutic measures. 25118277 2 80 theme poor 435:438 arg1 prognosis 440:448 poor prognosis 435:448 poor prognosis 435:448 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 0 81 theme oncogenic 63:71 arg1 features 73:80 oncogenic features 63:80 oncogenic features 63:80 Immature truncated O-glycophenotype of cancer directly induces oncogenic features. 25118277 1 82 theme adenocarcinomas 325:339 arg1 development 310:320 the development 306:320 the development of adenocarcinomas 306:339 Aberrant expression of immature truncated O-glycans is a characteristic feature observed on virtually all epithelial cancer cells, and a very high frequency is observed in early epithelial premalignant lesions that precede the development of adenocarcinomas. 25118277 5 83 theme pancreatic 1121:1130 arg1 cancers 1132:1138 primary and metastatic pancreatic cancers 1098:1138 primary and metastatic pancreatic cancers 1098:1138 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 2 84 theme truncated 360:368 arg1 Tn 390:391 Tn 390:391 Tn 390:391 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 84 theme truncated 360:368 arg1 structures 379:388 the truncated O-glycan structures Tn and sialyl-Tn 356:405 the truncated O-glycan structures Tn and sialyl-Tn 356:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 2 84 theme truncated 360:368 arg1 sialyl-Tn 397:405 sialyl-Tn 397:405 sialyl-Tn 397:405 Expression of the truncated O-glycan structures Tn and sialyl-Tn is strongly associated with poor prognosis and overall low survival. 25118277 7 85 theme tissue 1608:1613 arg1 model 1615:1619 an organotypic tissue model 1593:1619 an organotypic tissue model 1593:1619 We next used gene editing to produce isogenic cell systems with and without homogenous truncated O-glycans that enabled, to our knowledge, the first polyomic and side-by-side evaluation of the cancer O-glycophenotype in an organotypic tissue model and in xenografts. 25118277 5 86 theme large 1082:1086 arg1 series 1088:1093 a large series 1080:1093 a large series of primary and metastatic pancreatic cancers to rule out somatic mutations 1080:1168 Here, we used exome sequencing of most glycosyltransferases in a large series of primary and metastatic pancreatic cancers to rule out somatic mutations as a cause of expression of truncated O-glycans. 25118277 3 87 theme glycosyltransferases 712:731 arg1 relocation 698:707 relocation 698:707 relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum 698:809 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 3 87 theme glycosyltransferases 712:731 arg1 dysregulation 612:624 dysregulation 612:624 dysregulation 612:624 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 3 87 theme glycosyltransferases 712:731 arg1 mutation 600:607 mutation 600:607 mutation 600:607 The genetic and biosynthetic mechanisms leading to accumulation of truncated O-glycans are not fully understood and include mutation or dysregulation of glycosyltransferases involved in elongation of O-glycans, as well as relocation of glycosyltransferases controlling initiation of O-glycosylation from Golgi to endoplasmic reticulum. 25118277 8 88 theme oncogenic 1715:1723 arg1 features 1725:1732 oncogenic features 1715:1732 oncogenic features of cell growth and invasion 1715:1760 The results strongly suggest that truncation of O-glycans directly induces oncogenic features of cell growth and invasion. 26515459 6 0 theme fungal 1280:1285 arg1 genus 1287:1291 a single fungal genus 1271:1291 a single fungal genus 1271:1291 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 1 1 from case 170:173 arg1 present 211:217 present 211:217 present 211:217 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 5 2 theme N-glycans 1068:1076 arg1 modification 1052:1063 the modification 1048:1063 the modification of N-glycans in fungi with zwitterionic moieties 1048:1112 These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. 26515459 0 3 theme Species 69:75 arg1 Comparison 43:52 An N-glycomic Comparison 29:52 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 0:76 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 26515459 2 4 attach released 310:317 arg1 pellets 368:374 proteolysed cell pellets 351:374 proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 351:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 2 4 attach released 310:317 arg2 glycans 302:308 the glycans 298:308 the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 298:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 6 5 theme single 1273:1278 arg1 genus 1287:1291 a single fungal genus 1271:1291 a single fungal genus 1271:1291 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 4 6 located present 963:969 arg2 galactofuranose 943:957 bisecting galactofuranose 933:957 bisecting galactofuranose 933:957 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 4 6 located present 963:969 arg1 manner 994:999 a species-dependent manner 974:999 a species-dependent manner 974:999 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 1 7 theme secreted 233:240 arg1 range 224:228 a range 222:228 a range of secreted and cell wall proteins 222:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 5 8 from modification 1052:1063 arg1 fungi 1081:1085 fungi 1081:1085 fungi with zwitterionic moieties 1081:1112 These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. 26515459 6 9 theme spectrometric 1164:1176 arg1 screening 1178:1186 mere mass spectrometric screening 1154:1186 mere mass spectrometric screening 1154:1186 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 3 10 theme fragmentation 648:660 arg1 patterns 662:669 MS/MS fragmentation patterns 642:669 MS/MS fragmentation patterns 642:669 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 6 11 theme mass 1159:1162 arg1 screening 1178:1186 mere mass spectrometric screening 1154:1186 mere mass spectrometric screening 1154:1186 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 3 12 theme major 503:507 arg1 all 524:526 all 524:526 all 524:526 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 3 12 theme major 503:507 arg1 structures 509:518 the major structures 499:518 the major structures 499:518 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 4 13 theme phosphoethanolamine 849:867 arg1 residues 869:876 phosphoethanolamine residues 849:876 phosphoethanolamine residues 849:876 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 1 14 theme cell 246:249 arg1 proteins 256:263 cell wall proteins 246:263 cell wall proteins 246:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 4 15 theme oligomannosidic 789:803 arg1 glycans 805:811 the oligomannosidic glycans 785:811 the oligomannosidic glycans 785:811 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 6 16 theme mere 1154:1157 arg1 screening 1178:1186 mere mass spectrometric screening 1154:1186 mere mass spectrometric screening 1154:1186 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 4 17 theme mannosidase 748:758 arg1 treatments 760:769 mannosidase treatments 748:769 mannosidase treatments 748:769 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 4 18 theme species-dependent 976:992 arg1 manner 994:999 a species-dependent manner 974:999 a species-dependent manner 974:999 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 6 19 theme N-glycosylation 1243:1257 arg1 nature 1233:1238 the subtly complex nature 1214:1238 the subtly complex nature of N-glycosylation 1214:1257 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 2 20 theme cell 363:366 arg1 pellets 368:374 proteolysed cell pellets 351:374 proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 351:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 3 21 theme MS/MS 642:646 arg1 patterns 662:669 MS/MS fragmentation patterns 642:669 MS/MS fragmentation patterns 642:669 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 2 22 theme proteolysed 351:361 arg1 pellets 368:374 proteolysed cell pellets 351:374 proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 351:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 0 23 theme More 0:3 arg1 Oligomannose 15:26 More Than Just Oligomannose 0:26 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 0:76 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 26515459 1 24 theme wall 251:254 arg1 proteins 256:263 cell wall proteins 246:263 cell wall proteins 246:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 3 25 from differences 689:699 arg1 structure 715:723 the actual structure 704:723 the actual structure 704:723 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 1 26 theme filamentous 178:188 arg1 fungi 190:194 filamentous fungi 178:194 filamentous fungi 178:194 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 27 theme proteins 256:263 arg1 range 224:228 a range 222:228 a range of secreted and cell wall proteins 222:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 4 28 theme outer 882:886 arg1 mannosylation 909:921 outer chain och1-dependent mannosylation 882:921 outer chain och1-dependent mannosylation 882:921 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 3 29 theme range 539:543 arg1 HexNAc 554:559 the range Hex(5-11)HexNAc(2) 535:562 the range Hex(5-11)HexNAc(2) 535:562 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 6 30 theme complex 1225:1231 arg1 nature 1233:1238 the subtly complex nature 1214:1238 the subtly complex nature of N-glycosylation 1214:1257 Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. 26515459 1 31 theme fungi 190:194 arg1 case 170:173 the case 166:173 the case of filamentous fungi 166:194 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 32 theme essential 100:108 arg1 set 110:112 an essential set 97:112 an essential set of post-translational modifications of proteins 97:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 32 theme essential 100:108 arg1 modifications 136:148 post-translational modifications 117:148 post-translational modifications of proteins 117:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 32 theme essential 100:108 arg1 N-glycosylation 78:92 N-glycosylation 78:92 N-glycosylation 78:92 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 2 33 theme Penicillium 385:395 arg1 species 397:403 three Penicillium species 379:403 three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 379:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 3 34 from variations 595:604 arg1 chromatograms 624:636 reversed-phase chromatograms 609:636 reversed-phase chromatograms 609:636 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 3 34 from variations 595:604 arg1 patterns 662:669 MS/MS fragmentation patterns 642:669 MS/MS fragmentation patterns 642:669 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 4 35 theme och1-dependent 894:907 arg1 mannosylation 909:921 outer chain och1-dependent mannosylation 882:921 outer chain och1-dependent mannosylation 882:921 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 3 36 theme Hex 545:547 arg1 HexNAc 554:559 the range Hex(5-11)HexNAc(2) 535:562 the range Hex(5-11)HexNAc(2) 535:562 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 3 37 theme actual 708:713 arg1 structure 715:723 the actual structure 704:723 the actual structure 704:723 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 4 38 theme chain 888:892 arg1 mannosylation 909:921 outer chain och1-dependent mannosylation 882:921 outer chain och1-dependent mannosylation 882:921 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 1 39 attach present 211:217 arg2 N-glycans 197:205 N-glycans 197:205 N-glycans 197:205 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 39 attach present 211:217 arg1 case 170:173 the case 166:173 the case of filamentous fungi 166:194 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 39 attach present 211:217 arg1 range 224:228 a range 222:228 a range of secreted and cell wall proteins 222:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 4 40 dep only 822:825 arg1 not 818:820 not 818:820 not 818:820 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 4 41 from manner 994:999 arg1 present 963:969 present 963:969 present 963:969 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 3 42 theme differences 689:699 arg1 indicative 675:684 indicative 675:684 indicative 675:684 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 2 43 dep species 397:403 arg1 dierckxii 409:417 P. dierckxii 406:417 P. dierckxii 406:417 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 2 43 dep species 397:403 arg1 nordicum 423:430 P. nordicum 420:430 P. nordicum 420:430 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 2 43 dep species 397:403 arg1 verrucosum 439:448 P. verrucosum 436:448 P. verrucosum 436:448 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 0 44 theme N-glycomic 32:41 arg1 Comparison 43:52 An N-glycomic Comparison 29:52 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 0:76 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 26515459 5 45 theme zwitterionic 1092:1103 arg1 moieties 1105:1112 zwitterionic moieties 1092:1112 zwitterionic moieties 1092:1112 These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. 26515459 1 46 theme proteins 153:160 arg1 modifications 136:148 post-translational modifications 117:148 post-translational modifications of proteins 117:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 5 47 with fungi 1081:1085 arg1 moieties 1105:1112 zwitterionic moieties 1092:1112 zwitterionic moieties 1092:1112 These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. 26515459 1 48 from present 211:217 arg1 case 170:173 the case 166:173 the case of filamentous fungi 166:194 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 4 49 theme Hydrofluoric 726:737 arg1 acid 739:742 Hydrofluoric acid 726:742 Hydrofluoric acid 726:742 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 0 50 dep Oligomannose 15:26 arg1 Comparison 43:52 An N-glycomic Comparison 29:52 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 0:76 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 26515459 1 51 located present 211:217 arg2 N-glycans 197:205 N-glycans 197:205 N-glycans 197:205 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 51 located present 211:217 arg1 range 224:228 a range 222:228 a range of secreted and cell wall proteins 222:263 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 51 located present 211:217 arg1 case 170:173 the case 166:173 the case of filamentous fungi 166:194 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 52 theme post-translational 117:134 arg1 modifications 136:148 post-translational modifications 117:148 post-translational modifications of proteins 117:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 4 53 from present 963:969 arg1 manner 994:999 a species-dependent manner 974:999 a species-dependent manner 974:999 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 4 54 attach present 963:969 arg2 galactofuranose 943:957 bisecting galactofuranose 933:957 bisecting galactofuranose 933:957 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 4 54 attach present 963:969 arg1 manner 994:999 a species-dependent manner 974:999 a species-dependent manner 974:999 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 1 55 theme modifications 136:148 arg1 set 110:112 an essential set 97:112 an essential set of post-translational modifications of proteins 97:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 55 theme modifications 136:148 arg1 modifications 136:148 post-translational modifications 117:148 post-translational modifications of proteins 117:160 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 1 55 theme modifications 136:148 arg1 N-glycosylation 78:92 N-glycosylation 78:92 N-glycosylation 78:92 N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. 26515459 0 56 theme Penicillium 57:67 arg1 Species 69:75 Penicillium Species 57:75 Penicillium Species 57:75 More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species. 26515459 3 57 theme mass 576:579 arg1 spectrometry 581:592 mass spectrometry 576:592 mass spectrometry 576:592 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 2 58 theme species 397:403 arg1 pellets 368:374 proteolysed cell pellets 351:374 proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes) 351:487 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 26515459 4 59 theme bisecting 933:941 arg1 galactofuranose 943:957 bisecting galactofuranose 933:957 bisecting galactofuranose 933:957 Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. 26515459 3 60 theme reversed-phase 609:622 arg1 chromatograms 624:636 reversed-phase chromatograms 609:636 reversed-phase chromatograms 609:636 Although the major structures are all within the range Hex(5-11)HexNAc(2) as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. 26515459 2 61 theme peptide/N-glycosidase 322:342 arg1 F 344:344 peptide/N-glycosidase F 322:344 peptide/N-glycosidase F 322:344 In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). 24491843 4 0 theme C 504:504 arg1 spectroscopy 510:521 (13)C NMR spectroscopy 500:521 (13)C NMR spectroscopy 500:521 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 1 theme -β-D-Galp- 828:837 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 2 theme similar 174:180 arg1 structure 202:210 a similar O-antigen to type A structure 172:210 a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 172:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 3 theme following 587:595 arg1 structure 597:605 the following structure 583:605 the following structure 583:605 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 4 theme O-antigen 182:190 arg1 structure 202:210 a similar O-antigen to type A structure 172:210 a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 172:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 5 theme 1→5 839:841 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 6 theme linkage 444:450 arg1 analyses 452:459 linkage analyses 444:459 linkage analyses 444:459 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 7 with disaccharide 565:576 arg1 structure 597:605 the following structure 583:605 the following structure 583:605 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 8 dep disaccharide 565:576 arg1 revealed 671:678 revealed 671:678 revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure 671:787 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 8 dep disaccharide 565:576 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 9 theme -α-D-Galp- 813:822 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 10 theme Burkholderia 93:104 arg1 bacterium 142:150 a US isolated bacterium 128:150 a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 128:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 10 theme Burkholderia 93:104 arg1 E0147 119:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 11 theme NMR 649:651 arg1 spectroscopy 653:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 12 dep H 494:494 arg1 1 492:492 1 492:492 1 492:492 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 13 theme Glycosyl 419:426 arg1 composition 428:438 Glycosyl composition 419:438 Glycosyl composition 419:438 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 14 theme oklahomensis 106:117 arg1 bacterium 142:150 a US isolated bacterium 128:150 a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 128:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 14 theme oklahomensis 106:117 arg1 E0147 119:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 15 theme type 195:198 arg1 structure 202:210 a similar O-antigen to type A structure 172:210 a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 172:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 16 with conjunction 464:474 arg1 1D 481:482 1D 481:482 1D 481:482 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 17 theme 1→3 824:826 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 18 theme A 200:200 arg1 structure 202:210 a similar O-antigen to type A structure 172:210 a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 172:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 0 19 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of polysaccharides 0:45 Structural characterization of polysaccharides expressed by Burkholderia oklahomensis E0147. 24491843 4 20 theme repeating 555:563 arg1 disaccharide 565:576 a repeating disaccharide 553:576 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 21 theme exopolysaccharide 711:727 arg1 presence 684:691 the presence 680:691 the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure 680:787 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 22 theme n 647:647 arg1 spectroscopy 653:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 0 23 theme polysaccharides 31:45 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of polysaccharides 0:45 Structural characterization of polysaccharides expressed by Burkholderia oklahomensis E0147. 24491843 4 24 from composition 428:438 arg1 conjunction 464:474 conjunction 464:474 conjunction with 1D 464:482 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 25 dep isolated 133:140 arg1 US 130:131 US 130:131 US 130:131 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 26 theme -2OAc-β-D-Galp- 793:807 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 27 dep structure 202:210 arg1 to 192:193 to 192:193 to 192:193 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 28 theme pathogenic 226:235 arg1 species 237:243 the highly pathogenic species 215:243 the highly pathogenic species 215:243 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 28 theme pathogenic 226:235 arg1 pseudomallei 259:270 Burkholderia pseudomallei 246:270 Burkholderia pseudomallei 246:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 0 29 theme oklahomensis 73:84 arg1 E0147 86:90 Burkholderia oklahomensis E0147 60:90 Burkholderia oklahomensis E0147 60:90 Structural characterization of polysaccharides expressed by Burkholderia oklahomensis E0147. 24491843 4 30 theme -β-D-Kdop- 843:852 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 31 from analyses 452:459 arg1 conjunction 464:474 conjunction 464:474 conjunction with 1D 464:482 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 32 theme 1→4 809:811 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 33 theme isolated 133:140 arg1 bacterium 142:150 a US isolated bacterium 128:150 a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 128:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 1 33 theme isolated 133:140 arg1 E0147 119:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 93:123 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 0 34 theme Burkholderia 60:71 arg1 E0147 86:90 Burkholderia oklahomensis E0147 60:90 Burkholderia oklahomensis E0147 60:90 Structural characterization of polysaccharides expressed by Burkholderia oklahomensis E0147. 24491843 4 35 theme 2→ 854:855 arg1 n 857:857 [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 790:857 a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n 553:857 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 36 theme co-extracted 698:709 arg1 exopolysaccharide 711:727 a co-extracted exopolysaccharide 696:727 a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure 696:787 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 1 37 theme species 237:243 arg1 structure 202:210 a similar O-antigen to type A structure 172:210 a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei 172:270 Burkholderia oklahomensis E0147 is a US isolated bacterium believed to express a similar O-antigen to type A structure of the highly pathogenic species, Burkholderia pseudomallei. 24491843 4 38 theme 2D 488:489 arg1 H 494:494 2D (1)H 488:494 2D (1)H 488:494 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 39 theme -2OAc-α-L-6dTalp- 626:642 arg1 spectroscopy 653:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 40 theme NMR 506:508 arg1 spectroscopy 510:521 (13)C NMR spectroscopy 500:521 (13)C NMR spectroscopy 500:521 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 24491843 4 41 theme -β-D-Glcp- 611:620 arg1 spectroscopy 653:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy 608:664 Glycosyl composition and linkage analyses in conjunction with 1D and 2D (1)H and (13)C NMR spectroscopy showed that the O-antigen was a repeating disaccharide with the following structure: [3)-β-D-Glcp-(1→3)-2OAc-α-L-6dTalp-(1→]n NMR spectroscopy also revealed the presence of a co-extracted exopolysaccharide previously described in B. pseudomallei, with the structure: [3)-2OAc-β-D-Galp-(1→4)-α-D-Galp-(1→3)-β-D-Galp-(1→5)-β-D-Kdop-(2→]n. 27731363 5 0 theme α-glucosidases 940:953 arg1 RNAi 932:935 RNAi 932:935 RNAi of α-glucosidases involved in early N-glycan trimming and quality control 932:1009 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 4 1 theme modified 805:812 arg1 glycans 827:833 more extensively modified paucimannose glycans 788:833 more extensively modified paucimannose glycans 788:833 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 4 1 theme modified 805:812 arg1 ones 858:861 fucosylated ones 846:861 fucosylated ones 846:861 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 6 2 theme N-glycan 1122:1129 arg1 processing 1131:1140 N-glycan processing 1122:1140 N-glycan processing towards paucimannose structures 1122:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 2 3 from impact 373:378 arg1 metamorphosis 387:399 the metamorphosis 383:399 the metamorphosis 383:399 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 4 4 theme transcript 602:611 arg1 levels 613:618 The transcript levels 598:618 The transcript levels for genes encoding N-glycan processing enzymes 598:665 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 0 5 theme beetle 101:106 arg1 development 77:87 postembryonic development 63:87 postembryonic development of the pest beetle 63:106 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 1 6 from larva 175:179 arg1 transition 159:168 the transition 155:168 the transition from larva to adult 155:188 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 3 7 theme N-glycosylation 556:570 arg1 pathway 572:578 the protein N-glycosylation pathway 544:578 the protein N-glycosylation pathway 544:578 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 0 8 theme pest 96:99 arg1 beetle 101:106 the pest beetle 92:106 the pest beetle 92:106 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 7 9 theme N-glycan 1408:1415 arg1 processing 1417:1426 efficient N-glycan processing 1398:1426 efficient N-glycan processing 1398:1426 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 8 10 theme processes 1463:1471 arg1 approach 1499:1506 an effective new approach 1482:1506 an effective new approach for insect control 1482:1525 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 8 10 theme processes 1463:1471 arg1 disruption 1443:1452 disruption 1443:1452 disruption of these processes 1443:1471 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 3 11 from involvement 529:539 arg1 metamorphosis 583:595 metamorphosis 583:595 metamorphosis 583:595 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 3 12 theme pathway 572:578 arg1 involvement 529:539 the involvement 525:539 the involvement of the protein N-glycosylation pathway in metamorphosis 525:595 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 7 13 theme protein 1366:1372 arg1 attachment 1383:1392 protein N-glycan attachment 1366:1392 protein N-glycan attachment 1366:1392 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 7 14 theme N-glycan 1374:1381 arg1 attachment 1383:1392 protein N-glycan attachment 1366:1392 protein N-glycan attachment 1366:1392 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 5 15 theme attachment 885:894 arg1 Blockage 864:871 Blockage 864:871 Blockage of N-glycan attachment 864:894 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 1 16 gly N-glycosylated 244:257 arg1 proteins 259:266 N-glycosylated proteins 244:266 N-glycosylated proteins 244:266 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 2 17 theme posttranslational 301:317 arg1 modification 319:330 an important posttranslational modification 288:330 an important posttranslational modification that influences protein activity 288:363 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 2 17 theme posttranslational 301:317 arg1 N-glycosylation 269:283 N-glycosylation 269:283 N-glycosylation 269:283 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 8 18 theme new 1495:1497 arg1 approach 1499:1506 an effective new approach 1482:1506 an effective new approach for insect control 1482:1525 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 8 18 theme new 1495:1497 arg1 disruption 1443:1452 disruption 1443:1452 disruption of these processes 1443:1471 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 5 19 theme larval 908:913 arg1 mortality 915:923 larval mortality 908:923 larval mortality 908:923 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 6 20 theme pupal 1204:1208 arg1 formation 1220:1228 pupal appendage formation 1204:1228 pupal appendage formation 1204:1228 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 3 21 theme protein 548:554 arg1 pathway 572:578 the protein N-glycosylation pathway 544:578 the protein N-glycosylation pathway 544:578 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 6 22 theme paucimannose 1150:1161 arg1 structures 1163:1172 paucimannose structures 1150:1172 paucimannose structures 1150:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 6 23 theme multiple 1091:1098 arg1 genes 1100:1104 multiple genes 1091:1104 multiple genes responsible for N-glycan processing towards paucimannose structures 1091:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 3 24 theme flour 448:452 arg1 beetle 454:459 the red flour beetle 440:459 the red flour beetle 440:459 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 0 25 theme Protein 0:6 arg1 N-glycosylation 8:22 Protein N-glycosylation 0:22 Protein N-glycosylation 0:22 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 5 26 theme quality 995:1001 arg1 control 1003:1009 quality control 995:1009 quality control 995:1009 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 0 27 theme N-glycan 28:35 arg1 trimming 37:44 N-glycan trimming 28:44 N-glycan trimming 28:44 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 3 28 from study 516:520 arg1 involvement 529:539 the involvement 525:539 the involvement of the protein N-glycosylation pathway in metamorphosis 525:595 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 2 29 theme protein 348:354 arg1 activity 356:363 protein activity 348:363 protein activity 348:363 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 1 30 theme complete 201:208 arg1 reorganization 215:228 a complete body reorganization 199:228 a complete body reorganization 199:228 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 4 31 theme paucimannose 814:825 arg1 glycans 827:833 more extensively modified paucimannose glycans 788:833 more extensively modified paucimannose glycans 788:833 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 4 31 theme paucimannose 814:825 arg1 ones 858:861 fucosylated ones 846:861 fucosylated ones 846:861 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 8 32 theme effective 1485:1493 arg1 approach 1499:1506 an effective new approach 1482:1506 an effective new approach for insect control 1482:1525 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 8 32 theme effective 1485:1493 arg1 disruption 1443:1452 disruption 1443:1452 disruption of these processes 1443:1471 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 1 33 theme body 210:213 arg1 reorganization 215:228 a complete body reorganization 199:228 a complete body reorganization 199:228 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 7 34 theme efficient 1398:1406 arg1 processing 1417:1426 efficient N-glycan processing 1398:1426 efficient N-glycan processing 1398:1426 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 6 35 theme appendage 1210:1218 arg1 formation 1220:1228 pupal appendage formation 1204:1228 pupal appendage formation 1204:1228 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 6 36 theme responsible 1106:1116 arg1 genes 1100:1104 multiple genes 1091:1104 multiple genes responsible for N-glycan processing towards paucimannose structures 1091:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 4 37 theme developmental 690:702 arg1 stages 704:709 later developmental stages 684:709 later developmental stages 684:709 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 3 38 theme red 444:446 arg1 beetle 454:459 the red flour beetle 440:459 the red flour beetle 440:459 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 0 39 gly N-glycosylation 8:22 arg1 beetle 101:106 the pest beetle 92:106 the pest beetle 92:106 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 4 40 theme later 684:688 arg1 stages 704:709 later developmental stages 684:709 later developmental stages 684:709 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 3 41 used used 435:438 arg2 we 432:433 we 432:433 we 432:433 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 7 42 theme post-embryonic 1311:1324 arg1 development 1326:1336 insect post-embryonic development 1304:1336 insect post-embryonic development 1304:1336 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 6 43 theme novel 1189:1193 arg1 roles 1195:1199 their novel roles 1183:1199 their novel roles in pupal appendage formation and adult eclosion 1183:1247 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 3 44 theme comprehensive 502:514 arg1 study 516:520 a first comprehensive study 494:520 a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis 494:595 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 4 45 gly fucosylated 846:856 arg1 ones 858:861 fucosylated ones 846:861 fucosylated ones 846:861 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 8 46 theme insect 1512:1517 arg1 control 1519:1525 insect control 1512:1525 insect control 1512:1525 Consequently, disruption of these processes could be an effective new approach for insect control. 27731363 7 47 theme hormonal 1286:1293 arg1 control 1295:1301 hormonal control 1286:1301 hormonal control 1286:1301 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 5 48 theme early 967:971 arg1 trimming 982:989 early N-glycan trimming 967:989 early N-glycan trimming 967:989 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 1 49 theme holometabolous 132:145 arg1 insects 147:153 holometabolous insects 132:153 holometabolous insects 132:153 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 6 50 theme adult 1234:1238 arg1 eclosion 1240:1247 adult eclosion 1234:1247 adult eclosion 1234:1247 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 4 51 theme N-glycan 639:646 arg1 enzymes 659:665 N-glycan processing enzymes 639:665 N-glycan processing enzymes 639:665 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 2 52 theme important 291:299 arg1 modification 319:330 an important posttranslational modification 288:330 an important posttranslational modification that influences protein activity 288:363 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 2 52 theme important 291:299 arg1 N-glycosylation 269:283 N-glycosylation 269:283 N-glycosylation 269:283 N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. 27731363 4 53 theme glycans 827:833 arg1 enrichment 774:783 an enrichment 771:783 an enrichment of more extensively modified paucimannose glycans, including fucosylated ones 771:861 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 3 54 theme first 496:500 arg1 study 516:520 a first comprehensive study 494:520 a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis 494:595 Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. 27731363 5 55 theme N-glycan 973:980 arg1 trimming 982:989 early N-glycan trimming 967:989 early N-glycan trimming 967:989 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 7 56 theme insect 1304:1309 arg1 development 1326:1336 insect post-embryonic development 1304:1336 insect post-embryonic development 1304:1336 Our findings revealed that, next to hormonal control, insect post-embryonic development and metamorphosis depend on protein N-glycan attachment and efficient N-glycan processing. 27731363 6 57 theme genes 1100:1104 arg1 knockdown 1078:1086 simultaneous knockdown 1065:1086 simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures 1065:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 4 58 theme fucosylated 846:856 arg1 ones 858:861 fucosylated ones 846:861 fucosylated ones 846:861 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 5 59 theme N-glycan 876:883 arg1 attachment 885:894 N-glycan attachment 876:894 N-glycan attachment 876:894 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 4 60 from larva 741:745 arg1 transition 725:734 transition 725:734 transition from larva to adult 725:754 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27731363 1 61 theme N-glycosylated 244:257 arg1 proteins 259:266 N-glycosylated proteins 244:266 N-glycosylated proteins 244:266 In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. 27731363 0 62 theme postembryonic 63:75 arg1 development 77:87 postembryonic development 63:87 postembryonic development of the pest beetle 63:106 Protein N-glycosylation and N-glycan trimming are required for postembryonic development of the pest beetle Tribolium castaneum. 27731363 6 63 theme simultaneous 1065:1076 arg1 knockdown 1078:1086 simultaneous knockdown 1065:1086 simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures 1065:1172 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 6 64 from roles 1195:1199 arg1 eclosion 1240:1247 adult eclosion 1234:1247 adult eclosion 1234:1247 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 6 64 from roles 1195:1199 arg1 formation 1220:1228 pupal appendage formation 1204:1228 pupal appendage formation 1204:1228 Additionally, simultaneous knockdown of multiple genes responsible for N-glycan processing towards paucimannose structures revealed their novel roles in pupal appendage formation and adult eclosion. 27731363 5 65 theme pupa 1034:1037 arg1 transition 1039:1048 pupa transition 1034:1048 pupa transition 1034:1048 Blockage of N-glycan attachment resulted in larval mortality, while RNAi of α-glucosidases involved in early N-glycan trimming and quality control disrupted the larva to pupa transition. 27731363 4 66 theme processing 648:657 arg1 enzymes 659:665 N-glycan processing enzymes 639:665 N-glycan processing enzymes 639:665 The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones. 27797117 0 0 theme Polysaccharides 82:96 arg1 Library 24:30 a Library 22:30 a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 22:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 1 1 theme much 260:263 arg1 attention 265:273 much attention 260:273 much attention 260:273 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 0 2 theme Sulfate 74:80 arg1 Polysaccharides 82:96 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides 35:96 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 35:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 0 3 with Polysaccharides 82:96 arg1 Patterns 140:147 Different Sulfation and Fucosylation Patterns 103:147 Different Sulfation and Fucosylation Patterns 103:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 1 4 theme fCS 183:185 arg1 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 4 theme fCS 183:185 arg1 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 5 5 dep detailed 1044:1051 arg1 spectroscopic 1060:1072 spectroscopic 1060:1072 spectroscopic 1060:1072 The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. 27797117 0 6 theme Different 103:111 arg1 Patterns 140:147 Different Sulfation and Fucosylation Patterns 103:147 Different Sulfation and Fucosylation Patterns 103:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 6 7 theme structure-activity 1158:1175 arg1 relationship 1177:1188 structure-activity relationship 1158:1188 structure-activity relationship 1158:1188 These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date. 27797117 2 8 theme low 328:330 arg1 mass 342:345 a low molecular mass 326:345 a low molecular mass fCS polysaccharide 326:364 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 3 9 theme derivatives 718:728 arg1 library 680:686 a small library 672:686 a small library of fucosylated (and sulfated) derivatives thereof 672:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 4 10 theme reactions 819:827 arg1 combination 794:804 the different combination 780:804 the different combination of only five reactions 780:827 To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. 27797117 4 11 theme modular 754:760 arg1 approach 762:769 a modular approach 752:769 a modular approach based on the different combination of only five reactions 752:827 To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. 27797117 0 12 theme Sulfation 113:121 arg1 Patterns 140:147 Different Sulfation and Fucosylation Patterns 103:147 Different Sulfation and Fucosylation Patterns 103:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 2 13 theme antithrombotic 447:460 arg1 safer 481:485 safer 481:485 safer 481:485 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 2 13 theme antithrombotic 447:460 arg1 drug 462:465 an antithrombotic drug 444:465 an antithrombotic drug that would be safer and more effective than heparin 444:517 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 1 14 theme -a 187:188 arg1 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 14 theme -a 187:188 arg1 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 0 15 theme Modular 2:8 arg1 Approach 10:17 A Modular Approach 0:17 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 0:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 3 16 theme sulfated 708:715 arg1 derivatives 718:728 fucosylated (and sulfated) derivatives 691:728 fucosylated (and sulfated) derivatives thereof 691:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 6 17 dep wider 1134:1138 arg1 relationship 1177:1188 structure-activity relationship 1158:1188 structure-activity relationship 1158:1188 These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date. 27797117 3 18 theme drugs 555:559 arg1 use 533:535 the use 529:535 the use of animal sourced drugs 529:559 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 5 19 theme detailed 1044:1051 arg1 analysis 1074:1081 detailed 2D NMR spectroscopic analysis 1044:1081 detailed 2D NMR spectroscopic analysis 1044:1081 The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. 27797117 6 20 theme accurate 1149:1156 arg1 study 1190:1194 a wider and more accurate structure-activity relationship study 1132:1194 a wider and more accurate structure-activity relationship study with respect to those reported in literature to date 1132:1247 These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date. 27797117 1 21 theme biological 288:297 arg1 properties 299:308 its biological properties 284:308 its biological properties 284:308 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 3 22 theme animal 540:545 arg1 drugs 555:559 animal sourced drugs 540:559 animal sourced drugs 540:559 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 0 23 theme Fucosylation 127:138 arg1 Patterns 140:147 Different Sulfation and Fucosylation Patterns 103:147 Different Sulfation and Fucosylation Patterns 103:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 3 24 theme unsulfated 629:638 arg1 polysaccharide 652:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 3 25 theme sourced 547:553 arg1 drugs 555:559 animal sourced drugs 540:559 animal sourced drugs 540:559 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 4 26 theme key 923:925 arg1 step 927:930 the key step 919:930 the key step 919:930 To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. 27797117 3 27 theme chondroitin 640:650 arg1 polysaccharide 652:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 3 28 theme microbial 611:619 arg1 polysaccharide 652:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 4 29 theme different 784:792 arg1 combination 794:804 the different combination 780:804 the different combination of only five reactions 780:827 To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. 27797117 2 30 theme strong 404:409 arg1 candidate 411:419 a strong candidate 402:419 a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin 402:517 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 2 30 theme strong 404:409 arg1 polysaccharide 351:364 a low molecular mass fCS polysaccharide 326:364 a low molecular mass fCS polysaccharide 326:364 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 0 31 theme Semi-Synthetic 35:48 arg1 Polysaccharides 82:96 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides 35:96 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 35:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 3 32 gly fucosylated 691:701 arg1 derivatives 718:728 fucosylated (and sulfated) derivatives 691:728 fucosylated (and sulfated) derivatives thereof 691:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 3 33 theme sourced 621:627 arg1 polysaccharide 652:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 a microbial sourced unsulfated chondroitin polysaccharide 609:665 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 5 34 theme branches 1018:1025 arg1 patterns 978:985 sulfation patterns 968:985 sulfation patterns 968:985 The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. 27797117 6 35 theme semi-synthetic 1090:1103 arg1 polysaccharides 1105:1119 These semi-synthetic polysaccharides 1084:1119 These semi-synthetic polysaccharides 1084:1119 These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date. 27797117 2 36 theme fCS 347:349 arg1 candidate 411:419 a strong candidate 402:419 a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin 402:517 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 2 36 theme fCS 347:349 arg1 polysaccharide 351:364 a low molecular mass fCS polysaccharide 326:364 a low molecular mass fCS polysaccharide 326:364 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 2 37 theme drug 462:465 arg1 development 429:439 the development 425:439 the development of an antithrombotic drug that would be safer and more effective than heparin 425:517 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 4 38 theme unprecedented 858:870 arg1 polysaccharide 872:885 an almost unprecedented polysaccharide 848:885 an almost unprecedented polysaccharide branching by O-glycosylation as the key step 848:930 To this aim, a modular approach based on the different combination of only five reactions was employed, with an almost unprecedented polysaccharide branching by O-glycosylation as the key step. 27797117 2 39 theme mass 342:345 arg1 candidate 411:419 a strong candidate 402:419 a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin 402:517 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 2 39 theme mass 342:345 arg1 polysaccharide 351:364 a low molecular mass fCS polysaccharide 326:364 a low molecular mass fCS polysaccharide 326:364 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 3 40 theme small 674:678 arg1 library 680:686 a small library 672:686 a small library of fucosylated (and sulfated) derivatives thereof 672:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 5 41 theme fucose 1011:1016 arg1 branches 1018:1025 the fucose branches 1007:1025 the fucose branches 1007:1025 The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. 27797117 2 42 theme molecular 332:340 arg1 mass 342:345 a low molecular mass 326:345 a low molecular mass fCS polysaccharide 326:364 In particular, a low molecular mass fCS polysaccharide has very recently been suggested as a strong candidate for the development of an antithrombotic drug that would be safer and more effective than heparin. 27797117 1 43 theme sea 223:225 arg1 cucumbers-has 227:239 sea cucumbers-has 223:239 sea cucumbers-has 223:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 5 44 theme sulfation 968:976 arg1 patterns 978:985 sulfation patterns 968:985 sulfation patterns 968:985 The library was differentiated for sulfation patterns and/or positions of the fucose branches, as confirmed by detailed 2D NMR spectroscopic analysis. 27797117 3 45 theme polysaccharide 652:665 arg1 transformation 591:604 the chemical transformation 578:604 the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof 578:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 1 46 theme Fucosylated 150:160 arg1 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 46 theme Fucosylated 150:160 arg1 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 6 47 theme wider 1134:1138 arg1 study 1190:1194 a wider and more accurate structure-activity relationship study 1132:1194 a wider and more accurate structure-activity relationship study with respect to those reported in literature to date 1132:1247 These semi-synthetic polysaccharides will allow a wider and more accurate structure-activity relationship study with respect to those reported in literature to date. 27797117 0 48 theme Chondroitin 62:72 arg1 Sulfate 74:80 Fucosylated Chondroitin Sulfate 50:80 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 35:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 1 49 located found 214:218 arg1 cucumbers-has 227:239 sea cucumbers-has 223:239 sea cucumbers-has 223:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 49 located found 214:218 arg2 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 49 located found 214:218 arg2 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 3 50 theme chemical 582:589 arg1 transformation 591:604 the chemical transformation 578:604 the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof 578:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 1 51 theme chondroitin 162:172 arg1 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 51 theme chondroitin 162:172 arg1 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 0 52 theme Fucosylated 50:60 arg1 Sulfate 74:80 Fucosylated Chondroitin Sulfate 50:80 Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns 35:147 A Modular Approach to a Library of Semi-Synthetic Fucosylated Chondroitin Sulfate Polysaccharides with Different Sulfation and Fucosylation Patterns. 27797117 3 53 theme fucosylated 691:701 arg1 derivatives 718:728 fucosylated (and sulfated) derivatives 691:728 fucosylated (and sulfated) derivatives thereof 691:736 To avoid the use of animal sourced drugs, here we present the chemical transformation of a microbial sourced unsulfated chondroitin polysaccharide into a small library of fucosylated (and sulfated) derivatives thereof. 27797117 1 54 theme sulfate 174:180 arg1 glycosaminoglycan 190:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan 150:206 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has 150:239 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 27797117 1 54 theme sulfate 174:180 arg1 GAG 209:211 GAG 209:211 GAG 209:211 Fucosylated chondroitin sulfate (fCS)-a glycosaminoglycan (GAG) found in sea cucumbers-has recently attracted much attention owing to its biological properties. 25377922 1 0 theme important 194:202 arg1 products 212:219 many pharmaceutically important natural products 172:219 many pharmaceutically important natural products 172:219 2-Deoxy sugars and their derivatives occur abundantly in many pharmaceutically important natural products. 25377922 5 1 theme 2-deoxy-tetrasaccharide 755:777 arg1 synthesis 740:748 the synthesis 736:748 the synthesis of a 2-deoxy-tetrasaccharide containing four α-linkages 736:804 This method has also been applied to the synthesis of a 2-deoxy-tetrasaccharide containing four α-linkages. 25377922 0 2 link α-linked 29:36 arg1 glycosides 46:55 α-linked 2-deoxy glycosides 29:55 α-linked 2-deoxy glycosides 29:55 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 1 3 theme natural 204:210 arg1 products 212:219 many pharmaceutically important natural products 172:219 many pharmaceutically important natural products 172:219 2-Deoxy sugars and their derivatives occur abundantly in many pharmaceutically important natural products. 25377922 3 4 theme subsequent 424:433 arg1 deiodination 477:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 3 5 theme 2-iodo-glycosyl 396:410 arg1 acetate 412:418 2-iodo-glycosyl acetate 396:418 2-iodo-glycosyl acetate 396:418 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 3 6 theme acetate 412:418 arg1 glycosylation 379:391 glycosylation 379:391 glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination 379:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 4 7 theme mono- 599:603 arg1 synthesis 573:581 the synthesis 569:581 the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency 569:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 0 8 theme Stereoselective 0:14 arg1 synthesis 16:24 Stereoselective synthesis 0:24 Stereoselective synthesis of α-linked 2-deoxy glycosides 0:55 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 3 9 theme reductive 467:475 arg1 deiodination 477:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 1 10 theme 2-Deoxy 115:121 arg1 sugars 123:128 2-Deoxy sugars 115:128 2-Deoxy sugars 115:128 2-Deoxy sugars and their derivatives occur abundantly in many pharmaceutically important natural products. 25377922 3 11 theme deiodination 477:488 arg1 glycosylation 379:391 glycosylation 379:391 glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination 379:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 4 12 theme excellent 655:663 arg1 stereoselectivity 665:681 excellent stereoselectivity 655:681 excellent stereoselectivity 655:681 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 3 13 theme visible-light-mediated 435:456 arg1 deiodination 477:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 3 14 theme tin-free 458:465 arg1 deiodination 477:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 4 15 with mono- 599:603 arg1 efficiency 687:696 efficiency 687:696 efficiency 687:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 4 15 with mono- 599:603 arg1 stereoselectivity 665:681 excellent stereoselectivity 655:681 excellent stereoselectivity 655:681 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 0 16 theme glycosides 46:55 arg1 synthesis 16:24 Stereoselective synthesis 0:24 Stereoselective synthesis of α-linked 2-deoxy glycosides 0:55 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 0 17 theme 2-deoxy 38:44 arg1 glycosides 46:55 α-linked 2-deoxy glycosides 29:55 α-linked 2-deoxy glycosides 29:55 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 0 18 theme α-linked 29:36 arg1 glycosides 46:55 α-linked 2-deoxy glycosides 29:55 α-linked 2-deoxy glycosides 29:55 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 4 19 from strategy 557:564 arg1 synthesis 573:581 the synthesis 569:581 the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency 569:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 2 20 theme bonds 279:283 arg1 construction 235:246 the construction 231:246 the construction of specific 2-deoxy-glycosidic bonds 231:283 However, the construction of specific 2-deoxy-glycosidic bonds remains as a challenge. 25377922 4 21 theme pentadeoxysaccharides 628:648 arg1 synthesis 573:581 the synthesis 569:581 the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency 569:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 3 22 gly glycosylation 379:391 arg1 acetate 412:418 2-iodo-glycosyl acetate 396:418 2-iodo-glycosyl acetate 396:418 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 3 22 gly glycosylation 379:391 arg1 deiodination 477:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 subsequent visible-light-mediated tin-free reductive deiodination 424:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 2 23 theme specific 251:258 arg1 bonds 279:283 specific 2-deoxy-glycosidic bonds 251:283 specific 2-deoxy-glycosidic bonds 251:283 However, the construction of specific 2-deoxy-glycosidic bonds remains as a challenge. 25377922 2 24 theme 2-deoxy-glycosidic 260:277 arg1 bonds 279:283 specific 2-deoxy-glycosidic bonds 251:283 specific 2-deoxy-glycosidic bonds 251:283 However, the construction of specific 2-deoxy-glycosidic bonds remains as a challenge. 25377922 3 25 theme efficient 330:338 arg1 way 340:342 an efficient way 327:342 an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination 327:488 Herein, we report an efficient way to prepare 2-deoxy-α-glycosides by glycosylation of 2-iodo-glycosyl acetate and subsequent visible-light-mediated tin-free reductive deiodination. 25377922 4 26 with di- 606:608 arg1 efficiency 687:696 efficiency 687:696 efficiency 687:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 4 26 with di- 606:608 arg1 stereoselectivity 665:681 excellent stereoselectivity 655:681 excellent stereoselectivity 655:681 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 4 27 with pentadeoxysaccharides 628:648 arg1 efficiency 687:696 efficiency 687:696 efficiency 687:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 4 27 with pentadeoxysaccharides 628:648 arg1 stereoselectivity 665:681 excellent stereoselectivity 655:681 excellent stereoselectivity 655:681 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 5 28 contain containing 779:788 arg1 2-deoxy-tetrasaccharide 755:777 a 2-deoxy-tetrasaccharide 753:777 a 2-deoxy-tetrasaccharide containing four α-linkages 753:804 This method has also been applied to the synthesis of a 2-deoxy-tetrasaccharide containing four α-linkages. 25377922 5 28 contain containing 779:788 arg2 α-linkages 795:804 four α-linkages 790:804 four α-linkages 790:804 This method has also been applied to the synthesis of a 2-deoxy-tetrasaccharide containing four α-linkages. 25377922 0 29 theme reductive 91:99 arg1 deiodination 101:112 visible-light-mediated reductive deiodination 68:112 visible-light-mediated reductive deiodination 68:112 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 4 30 theme postglycosylational-deiodination 524:555 arg1 strategy 557:564 the postglycosylational-deiodination strategy 520:564 the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency 520:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 25377922 1 31 theme many 172:175 arg1 products 212:219 many pharmaceutically important natural products 172:219 many pharmaceutically important natural products 172:219 2-Deoxy sugars and their derivatives occur abundantly in many pharmaceutically important natural products. 25377922 0 32 theme visible-light-mediated 68:89 arg1 deiodination 101:112 visible-light-mediated reductive deiodination 68:112 visible-light-mediated reductive deiodination 68:112 Stereoselective synthesis of α-linked 2-deoxy glycosides enabled by visible-light-mediated reductive deiodination. 25377922 4 33 theme di- 606:608 arg1 synthesis 573:581 the synthesis 569:581 the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency 569:696 We have successfully applied the postglycosylational-deiodination strategy in the synthesis of more than 30 mono-, di-, tri-, tetra- and pentadeoxysaccharides with excellent stereoselectivity and efficiency. 24912705 2 0 theme acyl 434:437 arg1 donor 439:443 acyl donor 434:443 acyl donor 434:443 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 5 1 theme modified 724:731 arg1 protein 733:739 the modified protein 720:739 the modified protein 720:739 Circular dichroism analysis showed the modified protein possessed a decreased α-helix and β-structure. 24912705 1 2 theme crosslinking 189:200 arg1 impacts 146:152 The impacts 142:152 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein 142:253 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 0 3 theme protein 93:99 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 0 3 theme protein 93:99 arg1 modification 23:34 property modification 14:34 property modification 14:34 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 6 4 theme surface 825:831 arg1 hydrophobicity 833:846 lower surface hydrophobicity 819:846 lower surface hydrophobicity 819:846 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 0 5 theme microbial 114:122 arg1 transglutaminase 124:139 microbial transglutaminase 114:139 microbial transglutaminase 114:139 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 6 6 dep capacity 970:977 arg1 ml 994:995 12.2g and 3.5 ml 980:995 12.2g and 3.5 ml g(-1) protein 980:1009 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 3 7 theme modified 483:490 arg1 protein 492:498 The modified protein 479:498 The modified protein 479:498 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 6 8 theme binding 962:968 arg1 capacity 970:977 oil binding capacity 958:977 oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively) 958:1024 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 5 9 theme Circular 685:692 arg1 analysis 704:711 Circular dichroism analysis 685:711 Circular dichroism analysis 685:711 Circular dichroism analysis showed the modified protein possessed a decreased α-helix and β-structure. 24912705 6 10 theme emulsion 884:891 arg1 stability 893:901 higher emulsion stability 877:901 higher emulsion stability 877:901 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 2 11 from pH 352:353 arg1 protein 341:347 10 kU kg(-1) protein 328:347 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h 328:373 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 7 12 theme Transglutaminase-induced 1105:1128 arg1 glycosylation 1144:1156 Transglutaminase-induced oligochitosan glycosylation 1105:1156 Transglutaminase-induced oligochitosan glycosylation 1105:1156 Transglutaminase-induced oligochitosan glycosylation and crosslinking is thus able to modify soybean proteins. 24912705 6 13 theme oil 958:960 arg1 capacity 970:977 oil binding capacity 958:977 oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively) 958:1024 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 5 14 theme dichroism 694:702 arg1 analysis 704:711 Circular dichroism analysis 685:711 Circular dichroism analysis 685:711 Circular dichroism analysis showed the modified protein possessed a decreased α-helix and β-structure. 24912705 6 15 theme higher 877:882 arg1 stability 893:901 higher emulsion stability 877:901 higher emulsion stability 877:901 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 6 16 from droplets 1083:1090 arg1 emulsion 1095:1102 emulsion 1095:1102 emulsion 1095:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 4 17 theme spectroscopy 596:607 arg1 analysis 609:616 Electrophoresis and infrared spectroscopy analysis 567:616 Electrophoresis and infrared spectroscopy analysis 567:616 Electrophoresis and infrared spectroscopy analysis confirmed the modified protein to be crosslinked and glycosylated. 24912705 2 18 theme oligochitosan 448:460 arg1 acceptor 462:469 oligochitosan acceptor 448:469 oligochitosan acceptor of 1:3 448:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 6 19 dep ml 994:995 arg1 protein 1003:1009 g(-1) protein 997:1009 12.2g and 3.5 ml g(-1) protein 980:1009 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 4 20 theme infrared 587:594 arg1 spectroscopy 596:607 infrared spectroscopy 587:607 infrared spectroscopy 587:607 Electrophoresis and infrared spectroscopy analysis confirmed the modified protein to be crosslinked and glycosylated. 24912705 6 21 theme oil 1079:1081 arg1 droplets 1083:1090 the oil droplets 1075:1090 the oil droplets in emulsion 1075:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 6 22 theme modified 792:799 arg1 protein 801:807 The modified protein 788:807 The modified protein 788:807 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 3 23 contain contained 500:508 arg2 glucosamine 510:520 glucosamine 510:520 glucosamine 510:520 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 3 23 contain contained 500:508 arg1 protein 492:498 The modified protein 479:498 The modified protein 479:498 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 3 23 contain contained 500:508 arg2 protein 492:498 The modified protein 479:498 The modified protein 479:498 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 3 23 contain contained 500:508 arg1 concentration 527:539 a concentration 525:539 a concentration of 12.1 g kg(-1) protein 525:564 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 1 24 from impacts 146:152 arg1 properties 223:232 properties 223:232 properties 223:232 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 1 24 from impacts 146:152 arg1 structure 209:217 structure 209:217 structure 209:217 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 6 25 contain had 937:939 arg1 it 929:930 it 929:930 it 929:930 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 6 25 contain had 937:939 arg2 water 948:952 water 948:952 water 948:952 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 6 25 contain had 937:939 arg2 capacity 970:977 oil binding capacity 958:977 oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively) 958:1024 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 2 26 theme 1:3 474:476 arg1 acceptor 462:469 oligochitosan acceptor 448:469 oligochitosan acceptor of 1:3 448:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 2 27 theme gl 406:407 arg1 content 392:398 a protein content 382:398 a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3 382:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 6 28 from layer 1066:1070 arg1 droplets 1083:1090 the oil droplets 1075:1090 the oil droplets in emulsion 1075:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 6 29 theme emulsifying 852:862 arg1 activity 864:871 emulsifying activity 852:871 emulsifying activity 852:871 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 2 30 theme kU 331:332 arg1 protein 341:347 10 kU kg(-1) protein 328:347 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h 328:373 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 4 31 theme modified 632:639 arg1 protein 641:647 the modified protein 628:647 the modified protein 628:647 Electrophoresis and infrared spectroscopy analysis confirmed the modified protein to be crosslinked and glycosylated. 24912705 2 32 theme ratio 425:429 arg1 content 392:398 a protein content 382:398 a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3 382:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 5 33 dep showed 713:718 arg1 possessed 741:749 possessed 741:749 showed the modified protein possessed a decreased α-helix and β-structure 713:785 Circular dichroism analysis showed the modified protein possessed a decreased α-helix and β-structure. 24912705 7 34 theme soybean 1198:1204 arg1 proteins 1206:1213 soybean proteins 1198:1213 soybean proteins 1198:1213 Transglutaminase-induced oligochitosan glycosylation and crosslinking is thus able to modify soybean proteins. 24912705 4 35 theme Electrophoresis 567:581 arg1 analysis 609:616 Electrophoresis and infrared spectroscopy analysis 567:616 Electrophoresis and infrared spectroscopy analysis 567:616 Electrophoresis and infrared spectroscopy analysis confirmed the modified protein to be crosslinked and glycosylated. 24912705 1 36 theme soybean 239:245 arg1 protein 247:253 a soybean protein 237:253 a soybean protein 237:253 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 2 37 theme molar 419:423 arg1 ratio 425:429 a molar ratio 417:429 a molar ratio of acyl donor to oligochitosan acceptor of 1:3 417:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 4 38 gly glycosylated 671:682 arg1 protein 641:647 the modified protein 628:647 the modified protein 628:647 Electrophoresis and infrared spectroscopy analysis confirmed the modified protein to be crosslinked and glycosylated. 24912705 3 39 theme protein 558:564 arg1 concentration 527:539 a concentration 525:539 a concentration of 12.1 g kg(-1) protein 525:564 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 3 40 theme g 549:549 arg1 -1 554:555 -1 554:555 -1 554:555 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 3 40 theme g 549:549 arg1 kg 551:552 12.1 g kg 544:552 12.1 g kg(-1) protein 544:564 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 1 41 theme protein 247:253 arg1 properties 223:232 properties 223:232 properties 223:232 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 1 41 theme protein 247:253 arg1 structure 209:217 structure 209:217 structure 209:217 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 5 42 theme decreased 753:761 arg1 α-helix 763:769 a decreased α-helix 751:769 a decreased α-helix 751:769 Circular dichroism analysis showed the modified protein possessed a decreased α-helix and β-structure. 24912705 0 43 mod modification 23:34 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 0 43 mod modification 23:34 arg3 property 14:21 property modification 14:34 property modification 14:34 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 6 44 theme g 997:997 arg1 protein 1003:1009 g(-1) protein 997:1009 12.2g and 3.5 ml g(-1) protein 980:1009 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 2 45 theme donor 439:443 arg1 -1 409:410 -1 409:410 -1 409:410 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 2 45 theme donor 439:443 arg1 gl 406:407 40 gl 403:407 40 gl(-1) 403:411 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 2 45 theme donor 439:443 arg1 ratio 425:429 a molar ratio 417:429 a molar ratio of acyl donor to oligochitosan acceptor of 1:3 417:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 6 46 theme outer 1060:1064 arg1 layer 1066:1070 a thicker protein outer layer 1042:1070 a thicker protein outer layer in the oil droplets in emulsion 1042:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 0 47 theme oligochitosan-glycosylated 42:67 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 7 48 theme oligochitosan 1130:1142 arg1 glycosylation 1144:1156 Transglutaminase-induced oligochitosan glycosylation 1105:1156 Transglutaminase-induced oligochitosan glycosylation 1105:1156 Transglutaminase-induced oligochitosan glycosylation and crosslinking is thus able to modify soybean proteins. 24912705 6 49 theme lower 819:823 arg1 hydrophobicity 833:846 lower surface hydrophobicity 819:846 lower surface hydrophobicity 819:846 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 1 50 dep structure 209:217 arg1 the 205:207 the 205:207 the 205:207 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 6 51 theme protein 1052:1058 arg1 layer 1066:1070 a thicker protein outer layer 1042:1070 a thicker protein outer layer in the oil droplets in emulsion 1042:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 2 52 from 37°C 363:366 arg1 protein 341:347 10 kU kg(-1) protein 328:347 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h 328:373 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 0 53 link crosslinked 73:83 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 6 54 theme thicker 1044:1050 arg1 layer 1066:1070 a thicker protein outer layer 1042:1070 a thicker protein outer layer in the oil droplets in emulsion 1042:1102 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 3 55 theme kg 551:552 arg1 protein 558:564 12.1 g kg(-1) protein 544:564 12.1 g kg(-1) protein 544:564 The modified protein contained glucosamine at a concentration of 12.1 g kg(-1) protein. 24912705 1 56 theme oligochitosan 157:169 arg1 glycosylation 171:183 oligochitosan glycosylation 157:183 oligochitosan glycosylation 157:183 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 0 57 theme soybean 85:91 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 2 58 theme protein 384:390 arg1 content 392:398 a protein content 382:398 a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3 382:476 The reaction was carried out by transglutaminase with 10 kU kg(-1) protein at pH 7.5 and 37°C for 3h, under a protein content of 40 gl(-1) and a molar ratio of acyl donor to oligochitosan acceptor of 1:3. 24912705 0 59 gly oligochitosan-glycosylated 42:67 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 1 60 theme glycosylation 171:183 arg1 impacts 146:152 The impacts 142:152 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein 142:253 The impacts of oligochitosan glycosylation and crosslinking on the structure and properties of a soybean protein were investigated. 24912705 0 61 theme crosslinked 73:83 arg1 protein 93:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 an oligochitosan-glycosylated and crosslinked soybean protein 39:99 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 24912705 6 62 theme soybean 912:918 arg1 protein 920:926 the soybean protein 908:926 the soybean protein 908:926 The modified protein exhibited lower surface hydrophobicity and emulsifying activity but higher emulsion stability than the soybean protein, it also had better water and oil binding capacity (12.2g and 3.5 ml g(-1) protein, respectively), and could form a thicker protein outer layer in the oil droplets in emulsion. 24912705 0 63 theme property 14:21 arg1 modification 23:34 property modification 14:34 property modification 14:34 Structure and property modification of an oligochitosan-glycosylated and crosslinked soybean protein generated by microbial transglutaminase. 27883907 0 0 theme solution 79:86 arg1 pattern 43:49 Bimodal concentration-dependent reactivity pattern 0:49 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: 0:70 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 0 0 theme solution 79:86 arg1 structure 88:96 the solution structure 75:96 the solution structure involved 75:105 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 3 1 theme donor 959:963 arg1 supramers 937:945 fundamentally different supramers 913:945 fundamentally different supramers of glycosyl donor, which have distinct chemical properties 913:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 3 2 theme structure 866:874 arg1 types 848:852 two types 844:852 two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 844:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 2 3 theme dilute 710:715 arg1 solutions 717:725 more dilute solutions 705:725 more dilute solutions 705:725 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 2 4 theme solution 485:492 arg1 rotation 469:476 the specific optical rotation 448:476 the specific optical rotation of the solution of 1 448:497 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 2 4 theme solution 485:492 arg1 parameters 425:434 these parameters 419:434 these parameters 419:434 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 5 theme arabinofuranosyl 153:168 arg1 1 177:177 1 177:177 1 177:177 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 5 theme arabinofuranosyl 153:168 arg1 donor 170:174 an arabinofuranosyl donor 150:174 an arabinofuranosyl donor (1) 150:178 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 6 theme more 705:708 arg1 solutions 717:725 more dilute solutions 705:725 more dilute solutions 705:725 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 7 dep formed 357:362 arg1 disaccharide 344:355 the disaccharide 340:355 the disaccharide formed (from 14% to 82%) 340:380 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 7 dep formed 357:362 arg1 % 379:379 14% to 82% 370:379 14% to 82% 370:379 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 8 with formation 552:560 arg1 structures 625:634 similar structures 617:634 similar structures 617:634 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 9 theme supramers 937:945 arg1 presence 901:908 the presence 897:908 the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 897:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 4 10 theme reactions 1115:1123 arg1 selectivity 1096:1106 selectivity 1096:1106 selectivity of the reactions in which they participate 1096:1149 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 4 10 theme reactions 1115:1123 arg1 look 1035:1038 a fresh look 1027:1038 a fresh look at the problems of reactivity of chemical compounds 1027:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 3 11 theme glycosyl 950:957 arg1 donor 959:963 glycosyl donor 950:963 glycosyl donor 950:963 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 1 12 theme 14 370:371 arg1 % 372:372 % 372:372 % 372:372 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 3 13 theme concentration 784:796 arg1 ranges 798:803 two concentration ranges 780:803 two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 780:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 4 14 theme fresh 1029:1033 arg1 look 1035:1038 a fresh look 1027:1038 a fresh look at the problems of reactivity of chemical compounds 1027:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 3 15 theme distinct 977:984 arg1 properties 995:1004 distinct chemical properties 977:1004 distinct chemical properties 977:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 1 16 theme donor 170:174 arg1 0.001-0.1 M 134:144 0.001-0.1 M 134:144 0.001-0.1 M 134:144 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 16 theme donor 170:174 arg1 concentration 119:131 concentration 119:131 concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) 119:178 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 3 17 theme solution 857:864 arg1 structure 866:874 solution structure 857:874 solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 857:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 2 18 theme reacting 565:572 arg1 supramers 583:591 supramers 583:591 supramers 583:591 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 2 18 theme reacting 565:572 arg1 species 574:580 reacting species 565:580 reacting species (supramers) of glycosyl donor 565:610 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 19 from days 317:320 arg1 yield 331:335 the yield 327:335 the yield of the disaccharide formed (from 14% to 82%) 327:380 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 19 from days 317:320 arg1 time 295:298 the reaction time 282:298 the reaction time (from 1.5 h to 3 days) 282:321 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 0 20 theme concentration-dependent 8:30 arg1 pattern 43:49 Bimodal concentration-dependent reactivity pattern 0:49 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: 0:70 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 0 20 theme concentration-dependent 8:30 arg1 structure 88:96 the solution structure 75:96 the solution structure involved 75:105 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 1 21 theme 82 377:378 arg1 % 372:372 % 372:372 % 372:372 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 22 theme similar 617:623 arg1 structures 625:634 similar structures 617:634 similar structures 617:634 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 4 23 theme chemical 1073:1080 arg1 compounds 1082:1090 chemical compounds 1073:1090 chemical compounds 1073:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 0 24 theme Bimodal 0:6 arg1 pattern 43:49 Bimodal concentration-dependent reactivity pattern 0:49 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: 0:70 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 0 24 theme Bimodal 0:6 arg1 structure 88:96 the solution structure 75:96 the solution structure involved 75:105 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 1 25 theme 1 249:249 arg1 activation 235:244 activation 235:244 activation of 1 235:249 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 26 theme 1 497:497 arg1 solution 485:492 the solution 481:492 the solution of 1 481:497 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 27 theme solutions 817:825 arg1 ranges 798:803 two concentration ranges 780:803 two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 780:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 0 28 theme reactivity 32:41 arg1 pattern 43:49 Bimodal concentration-dependent reactivity pattern 0:49 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: 0:70 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 0 28 theme reactivity 32:41 arg1 structure 88:96 the solution structure 75:96 the solution structure involved 75:105 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 3 29 theme reaction 808:815 arg1 solutions 817:825 reaction solutions 808:825 reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties 808:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 1 30 theme from -23 °C 259:269 arg1 to +7 °C 271:278 from -23 °C to +7 °C 259:278 from -23 °C to +7 °C 259:278 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 31 theme different 674:682 arg1 structures 625:634 similar structures 617:634 similar structures 617:634 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 32 theme found 732:736 arg1 0.01 M 762:767 0.01 M 762:767 0.01 M 762:767 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 3 32 theme found 732:736 arg1 concentration 747:759 The found critical concentration 728:759 The found critical concentration (0.01 M) 728:768 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 1 33 dep 3 315:315 arg1 to 312:313 to 312:313 to 312:313 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 0 34 theme donor 65:69 arg1 pattern 43:49 Bimodal concentration-dependent reactivity pattern 0:49 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: 0:70 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 0 34 theme donor 65:69 arg1 structure 88:96 the solution structure 75:96 the solution structure involved 75:105 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 4 35 theme reactivity 1059:1068 arg1 problems 1047:1054 the problems 1043:1054 the problems of reactivity of chemical compounds 1043:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 0 36 theme glycosyl 56:63 arg1 donor 65:69 a glycosyl donor 54:69 a glycosyl donor 54:69 Bimodal concentration-dependent reactivity pattern of a glycosyl donor: Is the solution structure involved? 27883907 2 37 theme optical 461:467 arg1 rotation 469:476 the specific optical rotation 448:476 the specific optical rotation of the solution of 1 448:497 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 38 theme chemical 986:993 arg1 properties 995:1004 distinct chemical properties 977:1004 distinct chemical properties 977:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 4 39 theme compounds 1082:1090 arg1 reactivity 1059:1068 reactivity 1059:1068 reactivity of chemical compounds 1059:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 3 40 contain have 972:975 arg1 donor 959:963 glycosyl donor 950:963 glycosyl donor 950:963 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 3 40 contain have 972:975 arg2 properties 995:1004 distinct chemical properties 977:1004 distinct chemical properties 977:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 2 41 theme specific 452:459 arg1 rotation 469:476 the specific optical rotation 448:476 the specific optical rotation of the solution of 1 448:497 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 42 theme different 927:935 arg1 supramers 937:945 fundamentally different supramers 913:945 fundamentally different supramers of glycosyl donor, which have distinct chemical properties 913:1004 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 2 43 theme species 574:580 arg1 formation 552:560 formation 552:560 formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions 552:725 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 44 from Changes 108:114 arg1 0.001-0.1 M 134:144 0.001-0.1 M 134:144 0.001-0.1 M 134:144 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 44 from Changes 108:114 arg1 concentration 119:131 concentration 119:131 concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) 119:178 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 45 from those 689:693 arg1 different 674:682 different 674:682 different 674:682 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 46 theme disaccharide 344:355 arg1 yield 331:335 the yield 327:335 the yield of the disaccharide formed (from 14% to 82%) 327:380 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 46 theme disaccharide 344:355 arg1 time 295:298 the reaction time 282:298 the reaction time (from 1.5 h to 3 days) 282:321 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 4 47 from problems 1047:1054 arg1 selectivity 1096:1106 selectivity 1096:1106 selectivity of the reactions in which they participate 1096:1149 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 4 47 from problems 1047:1054 arg1 look 1035:1038 a fresh look 1027:1038 a fresh look at the problems of reactivity of chemical compounds 1027:1090 These results allow a fresh look at the problems of reactivity of chemical compounds and selectivity of the reactions in which they participate. 27883907 2 48 dep structures 625:634 arg1 reactivities 643:654 reactivities 643:654 reactivities 643:654 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 2 49 theme donor 606:610 arg1 supramers 583:591 supramers 583:591 supramers 583:591 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 2 49 theme donor 606:610 arg1 species 574:580 reacting species 565:580 reacting species (supramers) of glycosyl donor 565:610 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 1 50 dep % 372:372 arg1 to 374:375 to 374:375 to 374:375 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 51 theme temperature 212:222 arg1 T 224:224 the temperature T 208:224 the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%) 208:380 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 1 52 theme reaction 286:293 arg1 time 295:298 the reaction time 282:298 the reaction time (from 1.5 h to 3 days) 282:321 Changes in concentration (0.001-0.1 M) of an arabinofuranosyl donor (1) have been shown to modulate the temperature T at which activation of 1 occurs (from -23 °C to +7 °C), the reaction time (from 1.5 h to 3 days) and the yield of the disaccharide formed (from 14% to 82%). 27883907 2 53 theme glycosyl 597:604 arg1 donor 606:610 glycosyl donor 597:610 glycosyl donor 597:610 At concentrations exceeding 0.01 M, these parameters, as well as the specific optical rotation of the solution of 1, virtually do not depend on concentration suggesting formation of reacting species (supramers) of glycosyl donor with similar structures, hence reactivities, but considerably different from those formed in more dilute solutions. 27883907 3 54 theme critical 738:745 arg1 0.01 M 762:767 0.01 M 762:767 0.01 M 762:767 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27883907 3 54 theme critical 738:745 arg1 concentration 747:759 The found critical concentration 728:759 The found critical concentration (0.01 M) 728:768 The found critical concentration (0.01 M) separates two concentration ranges of reaction solutions corresponding to two types of solution structure that are featured by the presence of fundamentally different supramers of glycosyl donor, which have distinct chemical properties. 27020775 0 0 theme multiple 95:102 arg1 digestions 111:120 multiple enzyme digestions 95:120 multiple enzyme digestions 95:120 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 3 1 theme lactoferrin 781:791 arg1 sequence 793:800 donkey lactoferrin sequence 774:800 donkey lactoferrin sequence 774:800 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 3 2 theme genomic 867:873 arg1 sequence 883:890 the horse genomic deduced sequence 857:890 the horse genomic deduced sequence (UniProtKB Acc 857:905 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 5 3 from positions 959:967 arg1 located 948:954 located 948:954 located 948:954 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 3 4 dep sequence 883:890 arg1 Acc 903:905 UniProtKB Acc 893:905 UniProtKB Acc 893:905 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 5 5 theme point 928:932 arg1 substitutions 934:946 five point substitutions 923:946 five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala) 923:1057 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 3 6 theme deduced 875:881 arg1 sequence 883:890 the horse genomic deduced sequence 857:890 the horse genomic deduced sequence (UniProtKB Acc 857:905 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 1 7 theme potential 358:366 arg1 allergen 373:380 a potential milk allergen 356:380 a potential milk allergen 356:380 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 6 8 gly glycosylated 1076:1087 arg1 protein 1089:1095 the glycosylated protein 1072:1095 the glycosylated protein 1072:1095 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 2 9 theme SPITC 526:530 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 9 theme SPITC 526:530 arg1 derivatization 532:545 SPITC derivatization 526:545 SPITC derivatization of tryptic digest 526:563 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 10 theme milk 368:371 arg1 allergen 373:380 a potential milk allergen 356:380 a potential milk allergen 356:380 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 5 11 dep positions 959:967 arg1 466 1006:1008 466 1006:1008 466 1006:1008 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 Asn → Ser 1028:1036 Asn → Ser 1028:1036 Asn → Ser 1028:1036 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 328 985:987 328 985:987 328 985:987 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 Ala → Gly 1011:1019 Ala → Gly 1011:1019 Ala → Gly 1011:1019 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 Thr → Ile/Leu 990:1002 Thr → Ile/Leu 990:1002 Thr → Ile/Leu 990:1002 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 91 969:970 91 969:970 91 969:970 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 668 1043:1045 668 1043:1045 668 1043:1045 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 642 1023:1025 642 1023:1025 642 1023:1025 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 Arg → His 973:981 Arg → His 973:981 Arg → His 973:981 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 5 11 dep positions 959:967 arg1 Ser → Ala 1048:1056 Ser → Ala 1048:1056 Ser → Ala 1048:1056 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 3 12 theme horse 861:865 arg1 sequence 883:890 the horse genomic deduced sequence 857:890 the horse genomic deduced sequence (UniProtKB Acc 857:905 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 0 13 theme mass 126:129 arg1 spectrometry 131:142 mass spectrometry 126:142 mass spectrometry 126:142 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 3 14 theme sequence 793:800 arg1 characterization 754:769 the almost complete characterization 734:769 the almost complete characterization 734:769 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 2 15 theme mass 682:685 arg1 spectrometry 687:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 15 theme mass 682:685 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 6 16 attach linked 1143:1148 arg2 glycans 1109:1115 glycans 1109:1115 glycans in donkey lactoferrin 1109:1137 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 16 attach linked 1143:1148 arg1 backbone 1165:1172 the protein backbone 1153:1172 the protein backbone 1153:1172 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 16 attach linked 1143:1148 arg3 bond 1187:1190 an amide bond 1178:1190 an amide bond to asparagine residues located at the positions 137, 281 and 476 1178:1255 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 2 17 theme desorption/ionization 660:680 arg1 spectrometry 687:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 17 theme desorption/ionization 660:680 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 18 attach isolated 387:394 arg2 protein 160:166 a protein 158:166 a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen 158:380 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 18 attach isolated 387:394 arg1 milk 408:411 donkey milk 401:411 donkey milk 401:411 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 18 attach isolated 387:394 arg2 Lactoferrin 145:155 Lactoferrin 145:155 Lactoferrin 145:155 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 3 19 theme donkey 774:779 arg1 sequence 793:800 donkey lactoferrin sequence 774:800 donkey lactoferrin sequence 774:800 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 2 20 theme laser 654:658 arg1 spectrometry 687:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 20 theme laser 654:658 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 21 theme enzymatic 504:512 arg1 electrospray 621:632 electrospray 621:632 electrospray 621:632 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 21 theme enzymatic 504:512 arg1 spectrometry 687:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 21 theme enzymatic 504:512 arg1 chromatography 605:618 reversed-phase high performance liquid chromatography 566:618 reversed-phase high performance liquid chromatography 566:618 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 21 theme enzymatic 504:512 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 21 theme enzymatic 504:512 arg1 derivatization 532:545 SPITC derivatization 526:545 SPITC derivatization of tryptic digest 526:563 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 22 theme biochemical 188:198 arg1 properties 200:209 biochemical properties 188:209 biochemical properties 188:209 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 22 theme biochemical 188:198 arg1 ability 254:260 iron-binding ability 241:260 iron-binding ability 241:260 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 22 theme biochemical 188:198 arg1 antibacterial 287:299 antibacterial 287:299 antibacterial 287:299 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 22 theme biochemical 188:198 arg1 activities 315:324 antiviral activities 305:324 antiviral activities 305:324 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 22 theme biochemical 188:198 arg1 immuno-modulation 222:238 immuno-modulation 222:238 immuno-modulation 222:238 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 1 22 theme biochemical 188:198 arg1 antioxidant 274:284 antioxidant 274:284 antioxidant 274:284 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 0 23 theme Sequence 0:7 arg1 characterization 9:24 Sequence characterization 0:24 Sequence characterization 0:24 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 2 24 theme matrix-assisted 638:652 arg1 spectrometry 687:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 matrix-assisted laser desorption/ionization mass spectrometry 638:698 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 24 theme matrix-assisted 638:652 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 6 25 theme donkey 1120:1125 arg1 lactoferrin 1127:1137 donkey lactoferrin 1120:1137 donkey lactoferrin 1120:1137 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 1 26 theme properties 200:209 arg1 array 179:183 an array 176:183 an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen 176:380 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 0 27 theme glycosylation 30:42 arg1 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 27 theme glycosylation 30:42 arg1 sites 44:48 glycosylation sites 30:48 glycosylation sites 30:48 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 6 28 theme amide 1181:1185 arg1 bond 1187:1190 an amide bond 1178:1190 an amide bond to asparagine residues located at the positions 137, 281 and 476 1178:1255 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 29 from glycans 1109:1115 arg1 lactoferrin 1127:1137 donkey lactoferrin 1120:1137 donkey lactoferrin 1120:1137 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 1 30 theme antiviral 305:313 arg1 activities 315:324 antiviral activities 305:324 antiviral activities 305:324 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 2 31 theme digest 558:563 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 31 theme digest 558:563 arg1 derivatization 532:545 SPITC derivatization 526:545 SPITC derivatization of tryptic digest 526:563 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 6 32 dep positions 1230:1238 arg1 137 1240:1242 137 1240:1242 137 1240:1242 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 32 dep positions 1230:1238 arg1 476 1253:1255 476 1253:1255 476 1253:1255 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 32 dep positions 1230:1238 arg1 281 1245:1247 281 1245:1247 281 1245:1247 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 2 33 theme primary 473:479 arg1 structure 481:489 its primary structure 469:489 its primary structure 469:489 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 34 theme tryptic 550:556 arg1 digest 558:563 tryptic digest 550:563 tryptic digest 550:563 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 6 35 theme asparagine 1195:1204 arg1 residues 1206:1213 asparagine residues 1195:1213 asparagine residues located at the positions 137, 281 and 476 1195:1255 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 2 36 theme structure 481:489 arg1 characterization 449:464 The characterization 445:464 The characterization 445:464 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 37 theme donkey 401:406 arg1 milk 408:411 donkey milk 401:411 donkey milk 401:411 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 0 38 theme donkey 68:73 arg1 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 3 39 theme covered 826:832 arg1 sequence 834:841 the covered sequence 822:841 the covered sequence 822:841 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 2 40 theme liquid 598:603 arg1 chromatography 605:618 reversed-phase high performance liquid chromatography 566:618 reversed-phase high performance liquid chromatography 566:618 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 40 theme liquid 598:603 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 41 theme performance 586:596 arg1 chromatography 605:618 reversed-phase high performance liquid chromatography 566:618 reversed-phase high performance liquid chromatography 566:618 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 41 theme performance 586:596 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 0 42 gly glycosylation 30:42 arg2 sites 44:48 glycosylation sites 30:48 glycosylation sites 30:48 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 42 gly glycosylation 30:42 arg2 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 42 gly glycosylation 30:42 arg1 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 5 43 theme located 948:954 arg1 substitutions 934:946 five point substitutions 923:946 five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala) 923:1057 O77811) by five point substitutions located at positions 91 (Arg → His), 328 (Thr → Ile/Leu), 466 (Ala → Gly), 642 (Asn → Ser) and 668 (Ser → Ala). 27020775 3 44 theme UniProtKB 893:901 arg1 Acc 903:905 UniProtKB Acc 893:905 UniProtKB Acc 893:905 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 2 45 theme high 581:584 arg1 chromatography 605:618 reversed-phase high performance liquid chromatography 566:618 reversed-phase high performance liquid chromatography 566:618 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 45 theme high 581:584 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 46 theme ion 416:418 arg1 chromatography 429:442 ion exchange chromatography 416:442 ion exchange chromatography 416:442 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 6 47 theme located 1215:1221 arg1 residues 1206:1213 asparagine residues 1195:1213 asparagine residues located at the positions 137, 281 and 476 1195:1255 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 48 theme protein 1089:1095 arg1 Analysis 1060:1067 Analysis 1060:1067 Analysis of the glycosylated protein 1060:1095 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 1 49 theme iron-binding 241:252 arg1 ability 254:260 iron-binding ability 241:260 iron-binding ability 241:260 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 2 50 theme reversed-phase 566:579 arg1 chromatography 605:618 reversed-phase high performance liquid chromatography 566:618 reversed-phase high performance liquid chromatography 566:618 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 2 50 theme reversed-phase 566:579 arg1 digestions 514:523 enzymatic digestions 504:523 enzymatic digestions 504:523 The characterization of its primary structure, by means of enzymatic digestions, SPITC derivatization of tryptic digest, reversed-phase high performance liquid chromatography, electrospray and matrix-assisted laser desorption/ionization mass spectrometry, is reported. 27020775 1 51 theme exchange 420:427 arg1 chromatography 429:442 ion exchange chromatography 416:442 ion exchange chromatography 416:442 Lactoferrin, a protein showing an array of biochemical properties, including immuno-modulation, iron-binding ability, as well as antioxidant, antibacterial and antiviral activities, but which may also represent a potential milk allergen, was isolated from donkey milk by ion exchange chromatography. 27020775 6 52 theme glycosylated 1076:1087 arg1 protein 1089:1095 the glycosylated protein 1072:1095 the glycosylated protein 1072:1095 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 6 53 from positions 1230:1238 arg1 located 1215:1221 located 1215:1221 located 1215:1221 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 0 54 theme lactoferrin 80:90 arg1 characterization 9:24 Sequence characterization 0:24 Sequence characterization 0:24 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 54 theme lactoferrin 80:90 arg1 sites 44:48 glycosylation sites 30:48 glycosylation sites 30:48 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 54 theme lactoferrin 80:90 arg1 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 55 dep characterization 9:24 arg1 identification 50:63 identification 50:63 identification 50:63 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 0 56 theme milk 75:78 arg1 lactoferrin 80:90 donkey milk lactoferrin 68:90 donkey milk lactoferrin 68:90 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 27020775 3 57 theme complete 745:752 arg1 characterization 754:769 the almost complete characterization 734:769 the almost complete characterization 734:769 Our results allowed the almost complete characterization of donkey lactoferrin sequence, that, at least for the covered sequence, differs from the horse genomic deduced sequence (UniProtKB Acc. 27020775 6 58 theme protein 1157:1163 arg1 backbone 1165:1172 the protein backbone 1153:1172 the protein backbone 1153:1172 Analysis of the glycosylated protein showed that glycans in donkey lactoferrin are linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. 27020775 0 59 theme enzyme 104:109 arg1 digestions 111:120 multiple enzyme digestions 95:120 multiple enzyme digestions 95:120 Sequence characterization and glycosylation sites identification of donkey milk lactoferrin by multiple enzyme digestions and mass spectrometry. 28400496 6 0 theme cell 1091:1094 arg1 components 1101:1110 cell wall components 1091:1110 cell wall components 1091:1110 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 7 1 theme integrated 1124:1133 arg1 approach 1148:1155 this integrated glycobiology approach 1119:1155 this integrated glycobiology approach 1119:1155 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 6 2 theme monosaccharide 834:847 arg1 determination 861:873 monosaccharide composition determination 834:873 monosaccharide composition determination 834:873 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 7 3 theme root 1256:1259 arg1 growth 1265:1270 root cap growth 1256:1270 root cap growth 1256:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 1 4 theme wall 238:241 arg1 components 243:252 cell wall components 233:252 cell wall components 233:252 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 9 5 theme epitopes 1660:1667 arg1 distribution 1620:1631 the polarized distribution 1606:1631 the polarized distribution of xyloglucan and extensin epitopes 1606:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 0 6 theme Pea 0:2 arg1 Maturation 16:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 6 7 theme immunocarbohydrate 793:810 arg1 profiling 823:831 immunocarbohydrate microarray profiling 793:831 immunocarbohydrate microarray profiling 793:831 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 1 8 theme body 163:166 arg1 support 127:133 support 127:133 support 127:133 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 1 8 theme body 163:166 arg1 protection 139:148 protection 139:148 protection 139:148 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 4 9 theme cell 577:580 arg1 release 528:534 release 528:534 release 528:534 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 9 theme cell 577:580 arg1 maturation 513:522 maturation 513:522 maturation 513:522 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 9 theme cell 577:580 arg1 event 593:597 an important but poorly characterized cell separation event 539:597 an important but poorly characterized cell separation event 539:597 Their maturation and release is an important but poorly characterized cell separation event. 28400496 5 10 theme cellular 642:649 arg1 dynamics 651:658 the complex cellular dynamics 630:658 the complex cellular dynamics underlying this process 630:682 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 2 11 theme specialized 263:273 arg1 cases 275:279 some specialized cases 258:279 some specialized cases 258:279 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 5 12 theme tip 758:760 arg1 walls 767:771 pea (Pisum sativum) root tip cell walls 733:771 pea (Pisum sativum) root tip cell walls 733:771 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 6 13 theme quantitative 924:935 arg1 transcription-PCR 945:961 quantitative reverse transcription-PCR 924:961 quantitative reverse transcription-PCR of cell wall biosynthetic genes 924:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 8 14 theme process 1428:1434 arg1 model 1404:1408 a model 1402:1408 a model of the separation process 1402:1434 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 9 15 theme adhesion 1461:1468 arg1 loss 1453:1456 loss 1453:1456 loss of adhesion by the dissolution of homogalacturonan in the middle lamellae 1453:1530 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 1 16 theme cells 108:112 arg1 vital 117:121 vital 117:121 vital 117:121 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 1 16 theme cells 108:112 arg1 adhesion 90:97 The adhesion 86:97 The adhesion of plant cells 86:112 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 6 17 theme Fourier-transformed 876:894 arg1 microspectroscopy 905:921 Fourier-transformed infrared microspectroscopy 876:921 Fourier-transformed infrared microspectroscopy 876:921 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 0 18 theme Cell 55:58 arg1 Dynamics 76:83 Complex Cell Wall Structural Dynamics 47:83 Complex Cell Wall Structural Dynamics 47:83 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 0 19 theme Cell 11:14 arg1 Maturation 16:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 0 20 theme Structural 65:74 arg1 Dynamics 76:83 Complex Cell Wall Structural Dynamics 47:83 Complex Cell Wall Structural Dynamics 47:83 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 9 21 theme middle 1516:1521 arg1 lamellae 1523:1530 the middle lamellae 1512:1530 the middle lamellae 1512:1530 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 6 22 theme biosynthetic 976:987 arg1 genes 989:993 cell wall biosynthetic genes 966:993 cell wall biosynthetic genes 966:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 8 23 theme detail 1341:1346 arg1 level 1332:1336 a new level 1326:1336 a new level of detail about border cell maturation 1326:1375 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 7 24 theme structural 1206:1215 arg1 release 1280:1286 the release 1276:1286 the release of border cells 1276:1302 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 7 24 theme structural 1206:1215 arg1 modes 1187:1191 multiple novel modes 1172:1191 multiple novel modes of cell wall structural and compositional rearrangement during root cap growth 1172:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 6 25 theme cell 966:969 arg1 genes 989:993 cell wall biosynthetic genes 966:993 cell wall biosynthetic genes 966:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 8 26 theme border 1354:1359 arg1 maturation 1366:1375 border cell maturation 1354:1375 border cell maturation 1354:1375 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 2 27 theme border 349:354 arg1 examples 366:373 examples 366:373 examples of this 366:381 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 2 27 theme border 349:354 arg1 cells 356:360 root cap-derived border cells 332:360 root cap-derived border cells 332:360 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 0 28 theme Wall 60:63 arg1 Dynamics 76:83 Complex Cell Wall Structural Dynamics 47:83 Complex Cell Wall Structural Dynamics 47:83 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 2 29 theme root 332:335 arg1 examples 366:373 examples 366:373 examples of this 366:381 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 2 29 theme root 332:335 arg1 cells 356:360 root cap-derived border cells 332:360 root cap-derived border cells 332:360 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 5 30 theme Pisum 738:742 arg1 pea 733:735 pea 733:735 pea (Pisum sativum) root tip cell walls 733:771 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 5 30 theme Pisum 738:742 arg1 sativum 744:750 Pisum sativum 738:750 Pisum sativum 738:750 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 9 31 theme active 1551:1556 arg1 process 1570:1576 an active biophysical process 1548:1576 an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes 1548:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 7 32 theme multiple 1172:1179 arg1 modes 1187:1191 multiple novel modes 1172:1191 multiple novel modes of cell wall structural and compositional rearrangement during root cap growth 1172:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 6 33 theme transmission 1031:1042 arg1 microscopy 1053:1062 transmission electron microscopy 1031:1062 transmission electron microscopy 1031:1062 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 9 34 theme cell 1581:1584 arg1 curvature 1586:1594 cell curvature 1581:1594 cell curvature 1581:1594 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 6 35 theme activities 1019:1028 arg1 determination 861:873 monosaccharide composition determination 834:873 monosaccharide composition determination 834:873 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 transcription-PCR 945:961 quantitative reverse transcription-PCR 924:961 quantitative reverse transcription-PCR of cell wall biosynthetic genes 924:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 profiling 823:831 immunocarbohydrate microarray profiling 793:831 immunocarbohydrate microarray profiling 793:831 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 analysis 996:1003 analysis 996:1003 analysis of hydrolytic activities 996:1028 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 microscopy 1053:1062 transmission electron microscopy 1031:1062 transmission electron microscopy 1031:1062 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 microspectroscopy 905:921 Fourier-transformed infrared microspectroscopy 876:921 Fourier-transformed infrared microspectroscopy 876:921 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 35 theme activities 1019:1028 arg1 immunolocalization 1069:1086 immunolocalization 1069:1086 immunolocalization of cell wall components 1069:1110 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 2 36 link cap-derived 337:347 arg1 examples 366:373 examples 366:373 examples of this 366:381 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 2 36 link cap-derived 337:347 arg1 cells 356:360 root cap-derived border cells 332:360 root cap-derived border cells 332:360 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 3 37 theme border-like 423:433 arg1 cells 435:439 border-like cells 423:439 border-like cells 423:439 Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. 28400496 5 38 theme pea 733:735 arg1 walls 767:771 pea (Pisum sativum) root tip cell walls 733:771 pea (Pisum sativum) root tip cell walls 733:771 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 7 39 theme border 1291:1296 arg1 cells 1298:1302 border cells 1291:1302 border cells 1291:1302 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 2 40 theme plant 290:294 arg1 cells 296:300 plant cells 290:300 plant cells 290:300 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 8 41 theme separation 1417:1426 arg1 process 1428:1434 the separation process 1413:1434 the separation process 1413:1434 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 0 42 theme Complex 47:53 arg1 Dynamics 76:83 Complex Cell Wall Structural Dynamics 47:83 Complex Cell Wall Structural Dynamics 47:83 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 6 43 theme wall 1096:1099 arg1 components 1101:1110 cell wall components 1091:1110 cell wall components 1091:1110 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 3 44 from cells 391:395 arg1 species 406:412 some species 401:412 some species known as border-like cells 401:439 Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. 28400496 1 45 theme associations 212:223 arg1 variety 191:197 a variety 189:197 a variety of molecular associations between cell wall components 189:252 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 1 45 theme associations 212:223 arg1 associations 212:223 molecular associations 202:223 molecular associations between cell wall components 202:252 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 6 46 theme composition 849:859 arg1 determination 861:873 monosaccharide composition determination 834:873 monosaccharide composition determination 834:873 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 9 47 theme xyloglucan 1636:1645 arg1 epitopes 1660:1667 xyloglucan and extensin epitopes 1636:1667 xyloglucan and extensin epitopes 1636:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 7 48 theme cap 1261:1263 arg1 growth 1265:1270 root cap growth 1256:1270 root cap growth 1256:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 1 49 theme cell 233:236 arg1 components 243:252 cell wall components 233:252 cell wall components 233:252 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 9 50 theme extensin 1651:1658 arg1 epitopes 1660:1667 xyloglucan and extensin epitopes 1636:1667 xyloglucan and extensin epitopes 1636:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 6 51 theme microarray 812:821 arg1 profiling 823:831 immunocarbohydrate microarray profiling 793:831 immunocarbohydrate microarray profiling 793:831 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 9 52 from dissolution 1477:1487 arg1 lamellae 1523:1530 the middle lamellae 1512:1530 the middle lamellae 1512:1530 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 7 53 theme compositional 1221:1233 arg1 rearrangement 1235:1247 cell wall structural and compositional rearrangement 1196:1247 rearrangement 1235:1247 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 1 54 theme plant 157:161 arg1 body 163:166 the plant body 153:166 the plant body 153:166 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 5 55 theme complex 634:640 arg1 dynamics 651:658 the complex cellular dynamics 630:658 the complex cellular dynamics underlying this process 630:682 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 5 56 theme root 753:756 arg1 walls 767:771 pea (Pisum sativum) root tip cell walls 733:771 pea (Pisum sativum) root tip cell walls 733:771 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 6 57 theme reverse 937:943 arg1 transcription-PCR 945:961 quantitative reverse transcription-PCR 924:961 quantitative reverse transcription-PCR of cell wall biosynthetic genes 924:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 4 58 theme characterized 563:575 arg1 release 528:534 release 528:534 release 528:534 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 58 theme characterized 563:575 arg1 maturation 513:522 maturation 513:522 maturation 513:522 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 58 theme characterized 563:575 arg1 event 593:597 an important but poorly characterized cell separation event 539:597 an important but poorly characterized cell separation event 539:597 Their maturation and release is an important but poorly characterized cell separation event. 28400496 5 59 theme cell 762:765 arg1 walls 767:771 pea (Pisum sativum) root tip cell walls 733:771 pea (Pisum sativum) root tip cell walls 733:771 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 4 60 theme separation 582:591 arg1 release 528:534 release 528:534 release 528:534 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 60 theme separation 582:591 arg1 maturation 513:522 maturation 513:522 maturation 513:522 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 60 theme separation 582:591 arg1 event 593:597 an important but poorly characterized cell separation event 539:597 an important but poorly characterized cell separation event 539:597 Their maturation and release is an important but poorly characterized cell separation event. 28400496 5 61 theme deeper 610:615 arg1 insight 617:623 a deeper insight 608:623 a deeper insight into the complex cellular dynamics underlying this process 608:682 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 1 62 theme plant 102:106 arg1 cells 108:112 plant cells 102:112 plant cells 102:112 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 28400496 6 63 theme infrared 896:903 arg1 microspectroscopy 905:921 Fourier-transformed infrared microspectroscopy 876:921 Fourier-transformed infrared microspectroscopy 876:921 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 64 theme hydrolytic 1008:1017 arg1 activities 1019:1028 hydrolytic activities 1008:1028 hydrolytic activities 1008:1028 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 7 65 theme rearrangement 1235:1247 arg1 release 1280:1286 the release 1276:1286 the release of border cells 1276:1302 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 7 65 theme rearrangement 1235:1247 arg1 modes 1187:1191 multiple novel modes 1172:1191 multiple novel modes of cell wall structural and compositional rearrangement during root cap growth 1172:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 9 66 theme homogalacturonan 1492:1507 arg1 dissolution 1477:1487 the dissolution 1473:1487 the dissolution of homogalacturonan in the middle lamellae 1473:1530 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 5 67 theme systematic 700:709 arg1 analysis 721:728 a systematic, detailed analysis 698:728 analysis 721:728 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 6 68 theme genes 989:993 arg1 determination 861:873 monosaccharide composition determination 834:873 monosaccharide composition determination 834:873 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 transcription-PCR 945:961 quantitative reverse transcription-PCR 924:961 quantitative reverse transcription-PCR of cell wall biosynthetic genes 924:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 profiling 823:831 immunocarbohydrate microarray profiling 793:831 immunocarbohydrate microarray profiling 793:831 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 analysis 996:1003 analysis 996:1003 analysis of hydrolytic activities 996:1028 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 microscopy 1053:1062 transmission electron microscopy 1031:1062 transmission electron microscopy 1031:1062 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 microspectroscopy 905:921 Fourier-transformed infrared microspectroscopy 876:921 Fourier-transformed infrared microspectroscopy 876:921 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 68 theme genes 989:993 arg1 immunolocalization 1069:1086 immunolocalization 1069:1086 immunolocalization of cell wall components 1069:1110 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 8 69 theme new 1328:1330 arg1 level 1332:1336 a new level 1326:1336 a new level of detail about border cell maturation 1326:1375 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 6 70 theme wall 971:974 arg1 genes 989:993 cell wall biosynthetic genes 966:993 cell wall biosynthetic genes 966:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 2 71 theme cap-derived 337:347 arg1 examples 366:373 examples 366:373 examples of this 366:381 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 2 71 theme cap-derived 337:347 arg1 cells 356:360 root cap-derived border cells 332:360 root cap-derived border cells 332:360 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 0 72 theme Border 4:9 arg1 Maturation 16:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation 0:25 Pea Border Cell Maturation and Release Involve Complex Cell Wall Structural Dynamics. 28400496 8 73 theme cell 1361:1364 arg1 maturation 1366:1375 border cell maturation 1354:1375 border cell maturation 1354:1375 Our findings provide a new level of detail about border cell maturation and enable us to develop a model of the separation process. 28400496 9 74 theme biophysical 1558:1568 arg1 process 1570:1576 an active biophysical process 1548:1576 an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes 1548:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 6 75 theme electron 1044:1051 arg1 microscopy 1053:1062 transmission electron microscopy 1031:1062 transmission electron microscopy 1031:1062 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 2 76 theme this 378:381 arg1 examples 366:373 examples 366:373 examples of this 366:381 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 2 76 theme this 378:381 arg1 cells 356:360 root cap-derived border cells 332:360 root cap-derived border cells 332:360 In some specialized cases, though, plant cells are programmed to detach, and root cap-derived border cells are examples of this. 28400496 5 77 theme detailed 712:719 arg1 analysis 721:728 a systematic, detailed analysis 698:728 analysis 721:728 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 7 78 theme cells 1298:1302 arg1 release 1280:1286 the release 1276:1286 the release of border cells 1276:1302 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 7 78 theme cells 1298:1302 arg1 modes 1187:1191 multiple novel modes 1172:1191 multiple novel modes of cell wall structural and compositional rearrangement during root cap growth 1172:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 3 79 theme Border 384:389 arg1 cells 391:395 Border cells 384:395 Border cells (in some species known as border-like cells) 384:440 Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. 28400496 7 80 theme glycobiology 1135:1146 arg1 approach 1148:1155 this integrated glycobiology approach 1119:1155 this integrated glycobiology approach 1119:1155 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 9 81 theme curvature 1586:1594 arg1 process 1570:1576 an active biophysical process 1548:1576 an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes 1548:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 4 82 theme important 542:550 arg1 release 528:534 release 528:534 release 528:534 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 82 theme important 542:550 arg1 maturation 513:522 maturation 513:522 maturation 513:522 Their maturation and release is an important but poorly characterized cell separation event. 28400496 4 82 theme important 542:550 arg1 event 593:597 an important but poorly characterized cell separation event 539:597 an important but poorly characterized cell separation event 539:597 Their maturation and release is an important but poorly characterized cell separation event. 28400496 3 83 theme expendable 453:462 arg1 barrier 464:470 an expendable barrier 450:470 an expendable barrier between roots and the environment 450:504 Border cells (in some species known as border-like cells) provide an expendable barrier between roots and the environment. 28400496 5 84 theme walls 767:771 arg1 analysis 721:728 a systematic, detailed analysis 698:728 analysis 721:728 To gain a deeper insight into the complex cellular dynamics underlying this process, we undertook a systematic, detailed analysis of pea (Pisum sativum) root tip cell walls. 28400496 9 85 theme polarized 1610:1618 arg1 distribution 1620:1631 the polarized distribution 1606:1631 the polarized distribution of xyloglucan and extensin epitopes 1606:1667 We propose that loss of adhesion by the dissolution of homogalacturonan in the middle lamellae is augmented by an active biophysical process of cell curvature driven by the polarized distribution of xyloglucan and extensin epitopes. 28400496 6 86 theme components 1101:1110 arg1 determination 861:873 monosaccharide composition determination 834:873 monosaccharide composition determination 834:873 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 transcription-PCR 945:961 quantitative reverse transcription-PCR 924:961 quantitative reverse transcription-PCR of cell wall biosynthetic genes 924:993 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 profiling 823:831 immunocarbohydrate microarray profiling 793:831 immunocarbohydrate microarray profiling 793:831 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 analysis 996:1003 analysis 996:1003 analysis of hydrolytic activities 996:1028 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 microscopy 1053:1062 transmission electron microscopy 1031:1062 transmission electron microscopy 1031:1062 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 microspectroscopy 905:921 Fourier-transformed infrared microspectroscopy 876:921 Fourier-transformed infrared microspectroscopy 876:921 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 6 86 theme components 1101:1110 arg1 immunolocalization 1069:1086 immunolocalization 1069:1086 immunolocalization of cell wall components 1069:1110 Our study included immunocarbohydrate microarray profiling, monosaccharide composition determination, Fourier-transformed infrared microspectroscopy, quantitative reverse transcription-PCR of cell wall biosynthetic genes, analysis of hydrolytic activities, transmission electron microscopy, and immunolocalization of cell wall components. 28400496 7 87 theme novel 1181:1185 arg1 modes 1187:1191 multiple novel modes 1172:1191 multiple novel modes of cell wall structural and compositional rearrangement during root cap growth 1172:1270 Using this integrated glycobiology approach, we identified multiple novel modes of cell wall structural and compositional rearrangement during root cap growth and the release of border cells. 28400496 1 88 theme molecular 202:210 arg1 associations 212:223 molecular associations 202:223 molecular associations between cell wall components 202:252 The adhesion of plant cells is vital for support and protection of the plant body and is maintained by a variety of molecular associations between cell wall components. 27503803 0 0 theme mass 83:86 arg1 spectrometry 88:99 mass spectrometry 83:99 mass spectrometry 83:99 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 5 1 theme anti-cancer 771:781 arg1 derivative 783:792 the putative anti-cancer derivative 758:792 the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 758:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 6 2 attach removing 1484:1491 arg2 enzyme 1477:1482 this enzyme 1472:1482 this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo 1472:1550 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 6 2 attach removing 1484:1491 arg1 in vivo 1544:1550 cancer-patient DBP in vivo 1525:1550 cancer-patient DBP in vivo 1525:1550 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 1 3 theme human 167:171 arg1 protein 191:197 human vitamin D binding protein 167:197 human vitamin D binding protein (DBP) 167:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 3 theme human 167:171 arg1 DBP 200:202 DBP 200:202 DBP 200:202 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 6 4 theme cancer-patient 1525:1538 arg1 in vivo 1544:1550 cancer-patient DBP in vivo 1525:1550 cancer-patient DBP in vivo 1525:1550 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 6 5 theme endoglycosidic 1323:1336 arg1 function 1338:1345 endoglycosidic function 1323:1345 endoglycosidic function 1323:1345 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 1 6 theme vitamin 173:179 arg1 protein 191:197 human vitamin D binding protein 167:197 human vitamin D binding protein (DBP) 167:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 6 theme vitamin 173:179 arg1 DBP 200:202 DBP 200:202 DBP 200:202 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 5 7 theme glycan 1082:1087 arg1 structure 1089:1097 the glycan structure 1078:1097 the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied 1078:1226 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 3 8 theme structural 361:370 arg1 evidence 372:379 glycan structural evidence 354:379 glycan structural evidence 354:379 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 9 theme glycan 398:403 arg1 analysis 413:420 glycan linkage analysis 398:420 glycan linkage analysis 398:420 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 5 10 theme combined 889:896 arg1 action 898:903 the combined action 885:903 the combined action of β-galactosidase and neuraminidase upon DBP 885:949 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 11 theme GcMAF 1109:1113 arg1 structure 1089:1097 the glycan structure 1078:1097 the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied 1078:1226 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 2 12 theme product 282:288 arg1 O-glycan 254:261 the O-glycan 250:261 the O-glycan of the Gc1S allele product 250:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 2 12 theme product 282:288 arg1 trisaccharide 333:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 1 13 attach attached 155:162 arg1 protein 191:197 human vitamin D binding protein 167:197 human vitamin D binding protein (DBP) 167:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 13 attach attached 155:162 arg2 structure 145:153 the O-glycan structure 132:153 the O-glycan structure attached to human vitamin D binding protein (DBP) 132:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 13 attach attached 155:162 arg1 DBP 200:202 DBP 200:202 DBP 200:202 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 2 14 theme allele 275:280 arg1 product 282:288 the Gc1S allele product 266:288 the Gc1S allele product 266:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 1 15 theme D 181:181 arg1 protein 191:197 human vitamin D binding protein 167:197 human vitamin D binding protein (DBP) 167:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 15 theme D 181:181 arg1 DBP 200:202 DBP 200:202 DBP 200:202 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 3 16 theme glycan 354:359 arg1 evidence 372:379 glycan structural evidence 354:379 glycan structural evidence 354:379 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 2 17 theme Gc1S 270:273 arg1 product 282:288 the Gc1S allele product 266:288 the Gc1S allele product 266:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 1 18 theme binding 183:189 arg1 protein 191:197 human vitamin D binding protein 167:197 human vitamin D binding protein (DBP) 167:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 1 18 theme binding 183:189 arg1 DBP 200:202 DBP 200:202 DBP 200:202 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 5 19 theme Gc 810:811 arg1 Factor 851:856 Gc Protein-derived Macrophage Activating Factor 810:856 Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 810:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 20 theme Gal-GalNAc-Thr 1120:1133 arg1 disaccharide 1135:1146 a Gal-GalNAc-Thr disaccharide 1118:1146 a Gal-GalNAc-Thr disaccharide 1118:1146 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 4 21 theme Gc1S 690:693 arg1 protein 695:701 the Gc1S protein 686:701 the Gc1S protein 686:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 0 22 theme Glycan 0:5 arg1 structure 7:15 Glycan structure 0:15 Glycan structure of Gc Protein-derived Macrophage Activating Factor 0:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 2 23 theme core 304:307 arg1 O-glycan 254:261 the O-glycan 250:261 the O-glycan of the Gc1S allele product 250:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 2 23 theme core 304:307 arg1 trisaccharide 333:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 3 24 theme serial 434:439 arg1 experiments 463:473 over 30 serial glycosidase-digestion experiments 426:473 over 30 serial glycosidase-digestion experiments 426:473 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 0 25 theme Gc 20:21 arg1 Factor 61:66 Gc Protein-derived Macrophage Activating Factor 20:66 Gc Protein-derived Macrophage Activating Factor 20:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 5 26 theme Protein-derived 813:827 arg1 Factor 851:856 Gc Protein-derived Macrophage Activating Factor 810:856 Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 810:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 27 theme active 1102:1107 arg1 GcMAF 1109:1113 active GcMAF 1102:1113 active GcMAF 1102:1113 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 0 28 link Protein-derived 23:37 arg1 Factor 61:66 Gc Protein-derived Macrophage Activating Factor 20:66 Gc Protein-derived Macrophage Activating Factor 20:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 2 29 theme linear 297:302 arg1 O-glycan 254:261 the O-glycan 250:261 the O-glycan of the Gc1S allele product 250:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 2 29 theme linear 297:302 arg1 trisaccharide 333:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 5 30 dep formed 875:880 arg1 GcMAF 859:863 GcMAF 859:863 GcMAF 859:863 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 4 31 theme same 651:654 arg1 O-glycan 613:620 the O-glycan 609:620 the O-glycan from the Gc1F protein 609:642 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 31 theme same 651:654 arg1 trisaccharide 663:675 the same linear trisaccharide 647:675 the same linear trisaccharide found on the Gc1S protein 647:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 5 32 theme Activating 840:849 arg1 Factor 851:856 Gc Protein-derived Macrophage Activating Factor 810:856 Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 810:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 3 33 theme mass 556:559 arg1 ESI-MS 575:580 ESI-MS 575:580 ESI-MS 575:580 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 33 theme mass 556:559 arg1 spectrometry 561:572 electrospray ionization mass spectrometry 532:572 electrospray ionization mass spectrometry (ESI-MS) 532:581 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 4 34 located found 677:681 arg1 protein 695:701 the Gc1S protein 686:701 the Gc1S protein 686:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 34 located found 677:681 arg2 trisaccharide 663:675 the same linear trisaccharide 647:675 the same linear trisaccharide found on the Gc1S protein 647:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 34 located found 677:681 arg2 O-glycan 613:620 the O-glycan 609:620 the O-glycan from the Gc1F protein 609:642 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 6 35 theme enzyme 1477:1482 arg1 possibility 1457:1467 the possibility 1453:1467 the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo 1453:1550 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 0 36 theme Macrophage 39:48 arg1 Factor 61:66 Gc Protein-derived Macrophage Activating Factor 20:66 Gc Protein-derived Macrophage Activating Factor 20:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 4 37 theme hexose 716:721 arg1 residue 723:729 the hexose residue 712:729 the hexose residue 712:729 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 37 theme hexose 716:721 arg1 galactose 734:742 galactose 734:742 galactose 734:742 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 5 38 theme E. coli 1013:1019 arg1 β-galactosidase 1021:1035 the activating E. coli β-galactosidase 998:1035 the activating E. coli β-galactosidase 998:1035 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 6 39 theme DBP 1368:1370 arg1 O-glycan 1372:1379 the DBP O-glycan 1364:1379 the DBP O-glycan 1364:1379 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 0 40 theme Protein-derived 23:37 arg1 Factor 61:66 Gc Protein-derived Macrophage Activating Factor 20:66 Gc Protein-derived Macrophage Activating Factor 20:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 6 41 theme O-glycan 1497:1504 arg1 trisaccharide 1506:1518 the O-glycan trisaccharide 1493:1518 the O-glycan trisaccharide 1493:1518 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 5 42 theme DBP 797:799 arg1 derivative 783:792 the putative anti-cancer derivative 758:792 the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 758:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 43 theme Macrophage 829:838 arg1 Factor 851:856 Gc Protein-derived Macrophage Activating Factor 810:856 Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 810:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 4 44 theme linear 656:661 arg1 O-glycan 613:620 the O-glycan 609:620 the O-glycan from the Gc1F protein 609:642 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 44 theme linear 656:661 arg1 trisaccharide 663:675 the same linear trisaccharide 647:675 the same linear trisaccharide found on the Gc1S protein 647:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 3 45 theme linkage 405:411 arg1 analysis 413:420 glycan linkage analysis 398:420 glycan linkage analysis 398:420 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 46 theme protein 521:527 arg1 analysis 498:505 analysis 498:505 analysis of the intact protein 498:527 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 47 theme glycosidase-digestion 441:461 arg1 experiments 463:473 over 30 serial glycosidase-digestion experiments 426:473 over 30 serial glycosidase-digestion experiments 426:473 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 6 48 theme digestion 1251:1259 arg1 results 1261:1267 glycosidase digestion results 1239:1267 glycosidase digestion results 1239:1267 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 5 49 theme β-galactosidase 908:922 arg1 action 898:903 the combined action 885:903 the combined action of β-galactosidase and neuraminidase upon DBP 885:949 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 0 50 theme Activating 50:59 arg1 Factor 61:66 Gc Protein-derived Macrophage Activating Factor 20:66 Gc Protein-derived Macrophage Activating Factor 20:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 2 51 theme reported 217:224 arg1 evidence 226:233 Previously reported evidence 206:233 Previously reported evidence 206:233 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 3 52 theme intact 514:519 arg1 protein 521:527 the intact protein 510:527 the intact protein 510:527 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 6 53 theme glycosidase 1239:1249 arg1 results 1261:1267 glycosidase digestion results 1239:1267 glycosidase digestion results 1239:1267 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 27503803 5 54 theme activating 1002:1011 arg1 β-galactosidase 1021:1035 the activating E. coli β-galactosidase 998:1035 the activating E. coli β-galactosidase 998:1035 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 4 55 from protein 636:642 arg1 O-glycan 613:620 the O-glycan 609:620 the O-glycan from the Gc1F protein 609:642 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 4 55 from protein 636:642 arg1 trisaccharide 663:675 the same linear trisaccharide 647:675 the same linear trisaccharide found on the Gc1S protein 647:701 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 1 56 theme O-glycan 136:143 arg1 structure 145:153 the O-glycan structure 132:153 the O-glycan structure attached to human vitamin D binding protein (DBP) 132:203 Disagreement exists regarding the O-glycan structure attached to human vitamin D binding protein (DBP). 27503803 4 57 theme Gc1F 631:634 arg1 protein 636:642 the Gc1F protein 627:642 the Gc1F protein 627:642 Results demonstrate that the O-glycan from the Gc1F protein is the same linear trisaccharide found on the Gc1S protein and that the hexose residue is galactose. 27503803 2 58 theme NeuNAc-Gal-GalNAc-Thr 311:331 arg1 O-glycan 254:261 the O-glycan 250:261 the O-glycan of the Gc1S allele product 250:288 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 2 58 theme NeuNAc-Gal-GalNAc-Thr 311:331 arg1 trisaccharide 333:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide 293:345 Previously reported evidence indicated that the O-glycan of the Gc1S allele product is the linear core 1 NeuNAc-Gal-GalNAc-Thr trisaccharide. 27503803 5 59 theme neuraminidase 928:940 arg1 action 898:903 the combined action 885:903 the combined action of β-galactosidase and neuraminidase upon DBP 885:949 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 60 from protein-leaving 1062:1076 arg1 nothing 1045:1051 nothing 1045:1051 nothing 1045:1051 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 5 61 dep Factor 851:856 arg1 formed 875:880 formed 875:880 is formed by the combined action of β-galactosidase and neuraminidase upon DBP 872:949 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 3 62 theme electrospray 532:543 arg1 ESI-MS 575:580 ESI-MS 575:580 ESI-MS 575:580 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 62 theme electrospray 532:543 arg1 spectrometry 561:572 electrospray ionization mass spectrometry 532:572 electrospray ionization mass spectrometry (ESI-MS) 532:581 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 5 63 theme putative 762:769 arg1 derivative 783:792 the putative anti-cancer derivative 758:792 the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 758:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 0 64 theme Factor 61:66 arg1 structure 7:15 Glycan structure 0:15 Glycan structure of Gc Protein-derived Macrophage Activating Factor 0:66 Glycan structure of Gc Protein-derived Macrophage Activating Factor as revealed by mass spectrometry. 27503803 5 65 link Protein-derived 813:827 arg1 Factor 851:856 Gc Protein-derived Macrophage Activating Factor 810:856 Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) 810:950 In addition, the putative anti-cancer derivative of DBP known as Gc Protein-derived Macrophage Activating Factor (GcMAF, which is formed by the combined action of β-galactosidase and neuraminidase upon DBP) was analyzed intact by ESI-MS, revealing that the activating E. coli β-galactosidase cleaves nothing from the protein-leaving the glycan structure of active GcMAF as a Gal-GalNAc-Thr disaccharide, regardless of the order in which β-galactosidase and neuraminidase are applied. 27503803 3 66 theme ionization 545:554 arg1 ESI-MS 575:580 ESI-MS 575:580 ESI-MS 575:580 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 3 66 theme ionization 545:554 arg1 spectrometry 561:572 electrospray ionization mass spectrometry 532:572 electrospray ionization mass spectrometry (ESI-MS) 532:581 Here, glycan structural evidence is provided from glycan linkage analysis and over 30 serial glycosidase-digestion experiments which were followed by analysis of the intact protein by electrospray ionization mass spectrometry (ESI-MS). 27503803 6 67 theme DBP 1540:1542 arg1 in vivo 1544:1550 cancer-patient DBP in vivo 1525:1550 cancer-patient DBP in vivo 1525:1550 Moreover, glycosidase digestion results show that α-N-Acetylgalactosamindase (nagalase) lacks endoglycosidic function and only cleaves the DBP O-glycan once it has been trimmed down to a GalNAc-Thr monosaccharide-precluding the possibility of this enzyme removing the O-glycan trisaccharide from cancer-patient DBP in vivo. 24797640 3 0 link β-linked 454:461 arg1 disaccharides 490:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 4 1 theme glucuronyl 839:848 arg1 disaccharides 850:862 glucuronyl disaccharides 839:862 glucuronyl disaccharides 839:862 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 6 2 theme syn 1103:1105 arg1 addition 1107:1114 the syn addition 1099:1114 the syn addition of 2-furylzinc bromide 1099:1137 In some cases, the α-epoxypentenosides could be successfully converted into terminal l-iduronic acids via the syn addition of 2-furylzinc bromide. 24797640 7 3 theme stereochemical 1222:1235 arg1 configuration 1237:1249 the stereochemical configuration 1218:1249 the stereochemical configuration of the terminal 4'-DP unit 1218:1276 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 3 4 theme DMDO 575:578 arg1 sequence 603:610 the DMDO oxidation/ring-opening sequence 571:610 the DMDO oxidation/ring-opening sequence 571:610 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 4 5 theme 4'-DP 626:630 arg1 disaccharides 632:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 3 6 theme oxidation/ring-opening 580:601 arg1 sequence 603:610 the DMDO oxidation/ring-opening sequence 571:610 the DMDO oxidation/ring-opening sequence 571:610 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 2 7 with ring-openings 327:339 arg1 nucleophiles 354:365 various nucleophiles 346:365 various nucleophiles 346:365 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 2 8 theme DMDO 292:295 arg1 oxidation 297:305 stereocontrolled DMDO oxidation 275:305 stereocontrolled DMDO oxidation 275:305 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 5 9 theme high 958:961 arg1 stereoselectivity 963:979 high stereoselectivity 958:979 high stereoselectivity using DMDO 958:990 Both α- and β-linked 4'-DP disaccharides could be epoxidized with high stereoselectivity using DMDO. 24797640 3 10 theme α- 447:448 arg1 disaccharides 490:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 4 11 theme α-linked 617:624 arg1 disaccharides 632:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 2 12 theme stereocontrolled 275:290 arg1 oxidation 297:305 stereocontrolled DMDO oxidation 275:305 stereocontrolled DMDO oxidation 275:305 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 7 13 theme oligosaccharide 1182:1196 arg1 synthesis 1198:1206 oligosaccharide synthesis 1182:1206 oligosaccharide synthesis 1182:1206 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 1 14 theme versatile 94:102 arg1 synthons 104:111 versatile synthons 94:111 versatile synthons for rare or higher-order pyranosides 94:148 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 1 14 theme versatile 94:102 arg1 4-Deoxypentenosides 62:80 4-Deoxypentenosides 62:80 4-Deoxypentenosides (4-DPs) 62:88 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 1 15 theme structural 181:190 arg1 diversification 192:206 structural diversification 181:206 structural diversification at the C5 position 181:225 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 5 16 theme α- 897:898 arg1 disaccharides 919:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides could be epoxidized with high stereoselectivity using DMDO. 24797640 6 17 theme l-iduronic 1078:1087 arg1 acids 1089:1093 terminal l-iduronic acids 1069:1093 terminal l-iduronic acids 1069:1093 In some cases, the α-epoxypentenosides could be successfully converted into terminal l-iduronic acids via the syn addition of 2-furylzinc bromide. 24797640 4 18 theme glycosyl 703:710 arg1 acceptors 712:720 glycosyl acceptors 703:720 glycosyl acceptors using BSP/Tf2O activation 703:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 2 19 theme syn 397:399 arg1 selectivity 401:411 syn selectivity 397:411 syn selectivity 397:411 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 7 20 theme unit 1273:1276 arg1 configuration 1237:1249 the stereochemical configuration 1218:1249 the stereochemical configuration of the terminal 4'-DP unit 1218:1276 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 6 21 theme terminal 1069:1076 arg1 acids 1089:1093 terminal l-iduronic acids 1069:1093 terminal l-iduronic acids 1069:1093 In some cases, the α-epoxypentenosides could be successfully converted into terminal l-iduronic acids via the syn addition of 2-furylzinc bromide. 24797640 2 22 theme epoxide 319:325 arg1 ring-openings 327:339 subsequent epoxide ring-openings 308:339 subsequent epoxide ring-openings with various nucleophiles 308:365 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 4 23 theme thiophenyl 675:684 arg1 donors 691:696 coupling thiophenyl 4-DP donors 666:696 coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation 666:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 5 24 theme β-linked 904:911 arg1 disaccharides 919:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides could be epoxidized with high stereoselectivity using DMDO. 24797640 4 25 theme disaccharides 850:862 arg1 elimination 824:834 the decarboxylative elimination 804:834 the decarboxylative elimination of glucuronyl disaccharides 804:862 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 1 26 theme rare 117:120 arg1 pyranosides 138:148 rare or higher-order pyranosides 117:148 rare or higher-order pyranosides 117:148 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 4 27 theme coupling 666:673 arg1 donors 691:696 coupling thiophenyl 4-DP donors 666:696 coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation 666:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 7 28 theme terminal 1258:1265 arg1 unit 1273:1276 the terminal 4'-DP unit 1254:1276 the terminal 4'-DP unit 1254:1276 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 4 29 theme β-linked 757:764 arg1 disaccharides 772:784 β-linked 4'-DP disaccharides 757:784 β-linked 4'-DP disaccharides 757:784 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 1 30 theme C5 215:216 arg1 position 218:225 the C5 position 211:225 the C5 position 211:225 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 3 31 theme β-linked 454:461 arg1 disaccharides 490:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 0 32 theme 4'-deoxypentenosyl 28:45 arg1 reactivity 14:23 reactivity 14:23 reactivity 14:23 Synthesis and reactivity of 4'-deoxypentenosyl disaccharides. 24797640 0 32 theme 4'-deoxypentenosyl 28:45 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis 0:8 Synthesis and reactivity of 4'-deoxypentenosyl disaccharides. 24797640 7 33 theme 4'-DP 1267:1271 arg1 unit 1273:1276 the terminal 4'-DP unit 1254:1276 the terminal 4'-DP unit 1254:1276 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 3 34 theme 4'-deoxypentenosyl 463:480 arg1 disaccharides 490:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 447:502 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 2 35 theme Previous 228:235 arg1 studies 237:243 Previous studies 228:243 Previous studies 228:243 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 1 36 from position 218:225 arg1 diversification 192:206 structural diversification 181:206 structural diversification at the C5 position 181:225 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 4 37 theme 4'-DP 766:770 arg1 disaccharides 772:784 β-linked 4'-DP disaccharides 757:784 β-linked 4'-DP disaccharides 757:784 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 7 38 theme novel 1164:1168 arg1 approach 1170:1177 a novel approach 1162:1177 a novel approach 1162:1177 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 7 39 theme post-glycosylative 1298:1315 arg1 stage 1317:1321 a post-glycosylative stage 1296:1321 a post-glycosylative stage 1296:1321 These studies support a novel approach to oligosaccharide synthesis, in which the stereochemical configuration of the terminal 4'-DP unit is established at a post-glycosylative stage. 24797640 4 40 with donors 691:696 arg1 acceptors 712:720 glycosyl acceptors 703:720 glycosyl acceptors using BSP/Tf2O activation 703:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 4 41 dep synthesized 651:661 arg1 whereas 749:755 whereas 749:755 whereas 749:755 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 1 42 theme higher-order 125:136 arg1 pyranosides 138:148 rare or higher-order pyranosides 117:148 rare or higher-order pyranosides 117:148 4-Deoxypentenosides (4-DPs) are versatile synthons for rare or higher-order pyranosides, and they provide an entry for structural diversification at the C5 position. 24797640 2 43 theme subsequent 308:317 arg1 ring-openings 327:339 subsequent epoxide ring-openings 308:339 subsequent epoxide ring-openings with various nucleophiles 308:365 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 4 44 link α-linked 617:624 arg1 disaccharides 632:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides 613:644 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 4 45 theme microwave 870:878 arg1 conditions 880:889 microwave conditions 870:889 microwave conditions 870:889 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 4 46 theme BSP/Tf2O 728:735 arg1 activation 737:746 BSP/Tf2O activation 728:746 BSP/Tf2O activation 728:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 4 47 theme decarboxylative 808:822 arg1 elimination 824:834 the decarboxylative elimination 804:834 the decarboxylative elimination of glucuronyl disaccharides 804:862 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 6 48 theme bromide 1131:1137 arg1 addition 1107:1114 the syn addition 1099:1114 the syn addition of 2-furylzinc bromide 1099:1137 In some cases, the α-epoxypentenosides could be successfully converted into terminal l-iduronic acids via the syn addition of 2-furylzinc bromide. 24797640 4 49 link β-linked 757:764 arg1 disaccharides 772:784 β-linked 4'-DP disaccharides 757:784 β-linked 4'-DP disaccharides 757:784 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 5 50 link β-linked 904:911 arg1 disaccharides 919:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides could be epoxidized with high stereoselectivity using DMDO. 24797640 2 51 theme various 346:352 arg1 nucleophiles 354:365 various nucleophiles 346:365 various nucleophiles 346:365 Previous studies have shown that 4-DPs undergo stereocontrolled DMDO oxidation; subsequent epoxide ring-openings with various nucleophiles can proceed with both anti or syn selectivity. 24797640 6 52 theme 2-furylzinc 1119:1129 arg1 bromide 1131:1137 2-furylzinc bromide 1119:1137 2-furylzinc bromide 1119:1137 In some cases, the α-epoxypentenosides could be successfully converted into terminal l-iduronic acids via the syn addition of 2-furylzinc bromide. 24797640 4 53 theme 4-DP 686:689 arg1 donors 691:696 coupling thiophenyl 4-DP donors 666:696 coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation 666:746 The α-linked 4'-DP disaccharides were synthesized by coupling thiophenyl 4-DP donors with glycosyl acceptors using BSP/Tf2O activation, whereas β-linked 4'-DP disaccharides were generated by the decarboxylative elimination of glucuronyl disaccharides under microwave conditions. 24797640 5 54 theme 4'-DP 913:917 arg1 disaccharides 919:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides 892:931 Both α- and β-linked 4'-DP disaccharides could be epoxidized with high stereoselectivity using DMDO. 24797640 3 55 theme disaccharides 490:502 arg1 synthesis 434:442 the synthesis 430:442 the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides 430:502 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 24797640 3 56 theme post-glycosylational 530:549 arg1 additions 555:563 their post-glycosylational C5' additions 524:563 their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence 524:610 Here, we report the synthesis of α- and β-linked 4'-deoxypentenosyl (4'-DP) disaccharides, and we investigate their post-glycosylational C5' additions using the DMDO oxidation/ring-opening sequence. 26745022 3 0 theme WM266-4 311:317 arg1 cells 319:323 WM266-4 cells 311:323 WM266-4 cells 311:323 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 5 1 theme important 796:804 arg1 structures 784:793 overexpressed β1,6-branched structures 756:793 overexpressed β1,6-branched structures 756:793 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 4 2 theme focal 598:602 arg1 adhesions 604:612 focal adhesions 598:612 focal adhesions of melanoma cells growing on VN 598:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 3 3 theme V 383:383 arg1 overexpression 330:343 overexpression 330:343 overexpression of β1,6-acetylglucosaminyltransferase V 330:383 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 3 4 theme β1,6-branched 401:413 arg1 N-glycans 415:423 β1,6-branched N-glycans 401:423 β1,6-branched N-glycans 401:423 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 2 5 theme ECM 291:293 arg1 proteins 295:302 ECM proteins 291:302 ECM proteins 291:302 N-glycosylation of integrins affects their interactions with ECM proteins. 26745022 5 6 theme overexpressed 756:768 arg1 structures 784:793 overexpressed β1,6-branched structures 756:793 overexpressed β1,6-branched structures 756:793 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 1 7 theme key 162:164 arg1 activator 166:174 a key activator 160:174 a key activator of the focal adhesion kinase (FAK) signaling pathway 160:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 7 theme key 162:164 arg1 binding 107:113 Integrin-dependent binding 88:113 Integrin-dependent binding of the cell to extracellular matrix (ECM) 88:155 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 3 8 theme β1,6-acetylglucosaminyltransferase 348:381 arg1 V 383:383 β1,6-acetylglucosaminyltransferase V 348:383 β1,6-acetylglucosaminyltransferase V 348:383 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 5 9 theme FAK 810:812 arg1 upregulation 814:825 FAK upregulation 810:825 FAK upregulation 810:825 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 1 10 theme Integrin-dependent 88:105 arg1 activator 166:174 a key activator 160:174 a key activator of the focal adhesion kinase (FAK) signaling pathway 160:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 10 theme Integrin-dependent 88:105 arg1 binding 107:113 Integrin-dependent binding 88:113 Integrin-dependent binding of the cell to extracellular matrix (ECM) 88:155 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 3 11 theme metastatic 470:479 arg1 cells 490:494 metastatic melanoma cells 470:494 metastatic melanoma cells 470:494 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 0 12 theme complex-type 14:25 arg1 N-glycans 27:35 β1,6-branched complex-type N-glycans 0:35 β1,6-branched complex-type N-glycans 0:35 β1,6-branched complex-type N-glycans affect FAK signaling in metastatic melanoma cells. 26745022 0 13 theme β1,6-branched 0:12 arg1 N-glycans 27:35 β1,6-branched complex-type N-glycans 0:35 β1,6-branched complex-type N-glycans 0:35 β1,6-branched complex-type N-glycans affect FAK signaling in metastatic melanoma cells. 26745022 4 14 theme integrin 578:585 arg1 co-localization 554:568 The co-localization 550:568 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN 550:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 3 15 from migration 519:527 arg1 VN 545:546 VN 545:546 VN 545:546 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 3 15 from migration 519:527 arg1 vitronectin 532:542 vitronectin 532:542 vitronectin (VN) 532:547 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 4 16 from interaction 662:672 arg1 transduction 684:695 signal transduction 677:695 signal transduction 677:695 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 3 17 theme FAK 463:465 arg1 phosphorylation 444:458 tyrosine phosphorylation 435:458 tyrosine phosphorylation of FAK 435:465 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 4 18 theme αvβ3 573:576 arg1 integrin 578:585 αvβ3 integrin 573:585 αvβ3 integrin 573:585 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 1 19 theme focal 183:187 arg1 FAK 206:208 FAK 206:208 FAK 206:208 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 19 theme focal 183:187 arg1 kinase 198:203 focal adhesion kinase 183:203 the focal adhesion kinase (FAK) signaling pathway 179:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 2 20 gly N-glycosylation 230:244 arg1 integrins 249:257 integrins 249:257 integrins 249:257 N-glycosylation of integrins affects their interactions with ECM proteins. 26745022 4 21 theme FAK 591:593 arg1 co-localization 554:568 The co-localization 550:568 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN 550:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 1 22 theme cell 122:125 arg1 activator 166:174 a key activator 160:174 a key activator of the focal adhesion kinase (FAK) signaling pathway 160:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 22 theme cell 122:125 arg1 binding 107:113 Integrin-dependent binding 88:113 Integrin-dependent binding of the cell to extracellular matrix (ECM) 88:155 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 23 theme adhesion 189:196 arg1 FAK 206:208 FAK 206:208 FAK 206:208 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 23 theme adhesion 189:196 arg1 kinase 198:203 focal adhesion kinase 183:203 the focal adhesion kinase (FAK) signaling pathway 179:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 3 24 theme tyrosine 435:442 arg1 phosphorylation 444:458 tyrosine phosphorylation 435:458 tyrosine phosphorylation of FAK 435:465 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 1 25 theme kinase 198:203 arg1 pathway 221:227 the focal adhesion kinase (FAK) signaling pathway 179:227 the focal adhesion kinase (FAK) signaling pathway 179:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 3 26 theme melanoma 481:488 arg1 cells 490:494 metastatic melanoma cells 470:494 metastatic melanoma cells 470:494 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 0 27 theme FAK 44:46 arg1 signaling 48:56 FAK signaling 44:56 FAK signaling 44:56 β1,6-branched complex-type N-glycans affect FAK signaling in metastatic melanoma cells. 26745022 4 28 from co-localization 554:568 arg1 adhesions 604:612 focal adhesions 598:612 focal adhesions of melanoma cells growing on VN 598:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 5 29 theme β1,6-branched 770:782 arg1 structures 784:793 overexpressed β1,6-branched structures 756:793 overexpressed β1,6-branched structures 756:793 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 0 30 theme metastatic 61:70 arg1 cells 81:85 metastatic melanoma cells 61:85 metastatic melanoma cells 61:85 β1,6-branched complex-type N-glycans affect FAK signaling in metastatic melanoma cells. 26745022 5 31 theme Melanoma 698:705 arg1 migration 712:720 Melanoma cell migration 698:720 Melanoma cell migration on VN 698:726 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 5 32 theme cell 707:710 arg1 migration 712:720 Melanoma cell migration 698:720 Melanoma cell migration on VN 698:726 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 1 33 theme extracellular 130:142 arg1 matrix 144:149 extracellular matrix 130:149 extracellular matrix (ECM) 130:155 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 33 theme extracellular 130:142 arg1 ECM 152:154 ECM 152:154 ECM 152:154 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 2 34 theme integrins 249:257 arg1 N-glycosylation 230:244 N-glycosylation 230:244 N-glycosylation of integrins 230:257 N-glycosylation of integrins affects their interactions with ECM proteins. 26745022 4 35 theme cells 626:630 arg1 adhesions 604:612 focal adhesions 598:612 focal adhesions of melanoma cells growing on VN 598:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 3 36 theme enhanced 510:517 arg1 migration 519:527 enhanced migration 510:527 enhanced migration on vitronectin (VN) 510:547 Using WM266-4 cells with overexpression of β1,6-acetylglucosaminyltransferase V, we showed that β1,6-branched N-glycans increased tyrosine phosphorylation of FAK in metastatic melanoma cells, resulting in enhanced migration on vitronectin (VN). 26745022 4 37 theme melanoma 617:624 arg1 cells 626:630 melanoma cells 617:630 melanoma cells growing on VN 617:644 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 4 38 theme signal 677:682 arg1 transduction 684:695 signal transduction 677:695 signal transduction 677:695 The co-localization of αvβ3 integrin and FAK in focal adhesions of melanoma cells growing on VN indicates their interaction in signal transduction. 26745022 1 39 theme signaling 211:219 arg1 pathway 221:227 the focal adhesion kinase (FAK) signaling pathway 179:227 the focal adhesion kinase (FAK) signaling pathway 179:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 0 40 theme melanoma 72:79 arg1 cells 81:85 metastatic melanoma cells 61:85 metastatic melanoma cells 61:85 β1,6-branched complex-type N-glycans affect FAK signaling in metastatic melanoma cells. 26745022 1 41 theme pathway 221:227 arg1 activator 166:174 a key activator 160:174 a key activator of the focal adhesion kinase (FAK) signaling pathway 160:227 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 1 41 theme pathway 221:227 arg1 binding 107:113 Integrin-dependent binding 88:113 Integrin-dependent binding of the cell to extracellular matrix (ECM) 88:155 Integrin-dependent binding of the cell to extracellular matrix (ECM) is a key activator of the focal adhesion kinase (FAK) signaling pathway. 26745022 5 42 from migration 712:720 arg1 VN 725:726 VN 725:726 VN 725:726 Melanoma cell migration on VN was mediated by αvβ3 caring overexpressed β1,6-branched structures, important for FAK upregulation. 26745022 2 43 with interactions 273:284 arg1 proteins 295:302 ECM proteins 291:302 ECM proteins 291:302 N-glycosylation of integrins affects their interactions with ECM proteins. 26631508 10 0 theme extended 1867:1874 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 3 1 theme disease 498:504 arg1 implications 467:478 the implications 463:478 the implications of this prevalent disease 463:504 Despite the implications of this prevalent disease, diagnosis currently takes years, partly due to the diversity in patient presentation. 26631508 6 2 from rheology 1233:1240 arg1 saliva 1189:1194 residual mucosal saliva 1172:1194 residual mucosal saliva 1172:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 7 3 theme staining 1419:1426 arg1 comparison 1369:1378 a comparison 1367:1378 a comparison of protein Western blotting and glycan staining 1367:1426 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 2 4 theme function 276:283 arg1 loss 259:262 This loss 254:262 This loss of salivary function 254:283 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 7 5 theme mucin 1454:1458 arg1 glycosylation 1460:1472 mucin glycosylation 1454:1472 mucin glycosylation 1454:1472 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 10 6 theme dryness 2063:2069 arg1 perception 2044:2053 the perception 2040:2053 the perception of oral dryness 2040:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 8 7 theme global 1701:1706 arg1 decline 1708:1714 a global decline 1699:1714 a global decline of charged glycans 1699:1733 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 5 8 theme index 860:864 arg1 dryness 844:850 dryness 844:850 dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva 844:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 8 theme index 860:864 arg1 questionnaires 866:879 bother index questionnaires 853:879 bother index questionnaires 853:879 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 6 9 theme significant 1147:1157 arg1 reduction 1159:1167 a statistically significant reduction 1131:1167 a statistically significant reduction in residual mucosal saliva 1131:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 10 10 theme residual 1928:1935 arg1 wetness 1945:1951 residual mucosal wetness 1928:1951 residual mucosal wetness 1928:1951 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 4 11 theme biological 617:626 arg1 fluid 628:632 a complicated biological fluid 603:632 a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7 603:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 11 theme biological 617:626 arg1 Saliva 593:598 Saliva 593:598 Saliva 593:598 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 11 12 from changes 2085:2091 arg1 glycosylation 2101:2113 MUC7 glycosylation 2096:2113 MUC7 glycosylation 2096:2113 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 8 13 theme glycans 1727:1733 arg1 decline 1708:1714 a global decline 1699:1714 a global decline of charged glycans 1699:1733 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 6 14 theme extensional 1221:1231 arg1 spinnbarkeit 1243:1254 spinnbarkeit 1243:1254 spinnbarkeit (stringiness) 1243:1268 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 6 14 theme extensional 1221:1231 arg1 rheology 1233:1240 extensional rheology 1221:1240 extensional rheology 1221:1240 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 11 15 theme MUC7 2096:2099 arg1 glycosylation 2101:2113 MUC7 glycosylation 2096:2113 MUC7 glycosylation 2096:2113 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 9 16 theme extended 1778:1785 arg1 structure 1807:1815 the extended core 2 disialylated structure 1774:1815 the extended core 2 disialylated structure 1774:1815 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 5 17 from dryness 844:850 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 17 from dryness 844:850 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 4 18 gly glycosylated 677:688 arg1 mucins 690:695 heavily glycosylated mucins 669:695 heavily glycosylated mucins MUC5B and MUC7 669:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 18 gly glycosylated 677:688 arg1 MUC7 707:710 MUC7 707:710 MUC7 707:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 18 gly glycosylated 677:688 arg1 MUC5B 697:701 MUC5B 697:701 MUC5B 697:701 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 5 19 theme mucosal 1021:1027 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 19 theme mucosal 1021:1027 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 20 from perception 830:839 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 20 from perception 830:839 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 21 theme protein 910:916 arg1 composition 918:928 protein composition 910:928 protein composition 910:928 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 6 22 theme flow 1206:1209 arg1 rate 1211:1214 salivary flow rate 1197:1214 salivary flow rate 1197:1214 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 6 23 from rate 1211:1214 arg1 saliva 1189:1194 residual mucosal saliva 1172:1194 residual mucosal saliva 1172:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 10 24 theme flow 1977:1980 arg1 rate 1982:1985 whole mouth saliva flow rate 1958:1985 whole mouth saliva flow rate 1958:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 8 25 from increase 1616:1623 arg1 sulfation 1635:1643 core 1 sulfation 1628:1643 core 1 sulfation 1628:1643 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 6 26 theme mucosal 1181:1187 arg1 saliva 1189:1194 residual mucosal saliva 1172:1194 residual mucosal saliva 1172:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 4 27 theme glycosylated 677:688 arg1 mucins 690:695 heavily glycosylated mucins 669:695 heavily glycosylated mucins MUC5B and MUC7 669:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 27 theme glycosylated 677:688 arg1 MUC7 707:710 MUC7 707:710 MUC7 707:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 27 theme glycosylated 677:688 arg1 MUC5B 697:701 MUC5B 697:701 MUC5B 697:701 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 10 28 theme mouth 1964:1968 arg1 flow 1977:1980 whole mouth saliva flow 1958:1980 whole mouth saliva flow rate 1958:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 0 29 theme Saliva 47:52 arg1 Rheology 54:61 Altered Saliva Rheology 39:61 Altered Saliva Rheology 39:61 Reduced Mucin-7 (Muc7) Sialylation and Altered Saliva Rheology in Sjögren's Syndrome Associated Oral Dryness. 26631508 10 30 gly fucosylated 1877:1887 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 1 31 theme chronic 134:140 arg1 disorder 153:160 a chronic autoimmune disorder 132:160 a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands 132:251 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 1 31 theme chronic 134:140 arg1 syndrome 120:127 Sjögren's syndrome 110:127 Sjögren's syndrome 110:127 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 4 32 dep mucins 690:695 arg1 mucins 690:695 heavily glycosylated mucins 669:695 heavily glycosylated mucins MUC5B and MUC7 669:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 32 dep mucins 690:695 arg1 MUC7 707:710 MUC7 707:710 MUC7 707:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 32 dep mucins 690:695 arg1 MUC5B 697:701 MUC5B 697:701 MUC5B 697:701 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 6 33 from reduction 1159:1167 arg1 saliva 1189:1194 residual mucosal saliva 1172:1194 residual mucosal saliva 1172:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 5 34 theme patient 1083:1089 arg1 wellbeing 1091:1099 patient wellbeing 1083:1099 patient wellbeing 1083:1099 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 9 35 gly disialylated 1794:1805 arg1 structure 1807:1815 the extended core 2 disialylated structure 1774:1815 the extended core 2 disialylated structure 1774:1815 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 0 36 theme Reduced 0:6 arg1 Sialylation 23:33 Reduced Mucin-7 (Muc7) Sialylation 0:33 Reduced Mucin-7 (Muc7) Sialylation 0:33 Reduced Mucin-7 (Muc7) Sialylation and Altered Saliva Rheology in Sjögren's Syndrome Associated Oral Dryness. 26631508 5 37 theme glycan 935:940 arg1 analysis 942:949 glycan analysis 935:949 glycan analysis of whole mouth saliva 935:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 2 38 theme increased 420:428 arg1 risk 430:433 an increased risk 417:433 an increased risk of certain cancers 417:452 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 8 39 theme O-glycans 1531:1539 arg1 analysis 1519:1526 LC-MS/MS analysis 1510:1526 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination 1510:1575 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 10 40 contain have 1999:2002 arg1 decrease 1851:1858 A decrease 1849:1858 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate 1849:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 40 contain have 1999:2002 arg2 effect 2030:2035 a negative and cumulative effect 2004:2035 a negative and cumulative effect on the perception of oral dryness 2004:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 8 41 from reduction 1662:1670 arg1 sialylation 1675:1685 sialylation 1675:1685 sialylation 1675:1685 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 7 42 theme mucins 1301:1306 arg1 similar 1328:1334 similar 1328:1334 similar 1328:1334 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 7 42 theme mucins 1301:1306 arg1 concentration 1284:1296 the concentration 1280:1296 the concentration of mucins MUC5B and MUC7 1280:1321 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 11 43 theme diagnostic 2136:2145 arg1 changes 2085:2091 The observed changes 2072:2091 The observed changes in MUC7 glycosylation 2072:2113 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 11 43 theme diagnostic 2136:2145 arg1 tool 2147:2150 a potential diagnostic tool 2124:2150 a potential diagnostic tool for saliva quality 2124:2169 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 2 44 theme autoimmune 393:402 arg1 diseases 404:411 other autoimmune diseases 387:411 other autoimmune diseases 387:411 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 5 45 theme bother 853:858 arg1 dryness 844:850 dryness 844:850 dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva 844:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 45 theme bother 853:858 arg1 questionnaires 866:879 bother index questionnaires 853:879 bother index questionnaires 853:879 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 2 46 theme cancers 446:452 arg1 risk 430:433 an increased risk 417:433 an increased risk of certain cancers 417:452 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 2 46 theme cancers 446:452 arg1 diseases 404:411 other autoimmune diseases 387:411 other autoimmune diseases 387:411 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 8 47 theme charged 1719:1725 arg1 glycans 1727:1733 charged glycans 1719:1733 charged glycans 1719:1733 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 5 48 theme residual 1012:1019 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 48 theme residual 1012:1019 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 10 49 theme cumulative 2019:2028 arg1 effect 2030:2035 a negative and cumulative effect 2004:2035 a negative and cumulative effect on the perception of oral dryness 2004:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 4 50 with fluid 628:632 arg1 mucins 690:695 heavily glycosylated mucins 669:695 heavily glycosylated mucins MUC5B and MUC7 669:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 50 with fluid 628:632 arg1 constituents 645:656 major constituents 639:656 major constituents 639:656 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 5 51 theme saliva 981:986 arg1 perception 830:839 Sjögren's patient's perception 810:839 Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) 810:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 10 52 theme negative 2006:2013 arg1 effect 2030:2035 a negative and cumulative effect 2004:2035 a negative and cumulative effect on the perception of oral dryness 2004:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 11 53 theme future 2204:2209 arg1 therapies 2211:2219 future therapies 2204:2219 future therapies for this multifactorial syndrome 2204:2252 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 9 54 theme structure 1807:1815 arg1 This 1736:1739 This 1736:1739 This 1736:1739 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 9 54 theme structure 1807:1815 arg1 loss 1766:1769 the loss 1762:1769 the loss of the extended core 2 disialylated structure 1762:1815 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 7 55 dep mucins 1301:1306 arg1 mucins 1301:1306 mucins MUC5B and MUC7 1301:1321 mucins MUC5B and MUC7 1301:1321 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 7 55 dep mucins 1301:1306 arg1 MUC7 1318:1321 MUC7 1318:1321 MUC7 1318:1321 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 7 55 dep mucins 1301:1306 arg1 MUC5B 1308:1312 MUC5B 1308:1312 MUC5B 1308:1312 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 3 56 theme prevalent 488:496 arg1 disease 498:504 this prevalent disease 483:504 this prevalent disease 483:504 Despite the implications of this prevalent disease, diagnosis currently takes years, partly due to the diversity in patient presentation. 26631508 8 57 theme larger 1655:1660 arg1 reduction 1662:1670 the even larger reduction 1646:1670 the even larger reduction in sialylation 1646:1685 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 1 58 theme glands 246:251 arg1 infiltration 191:202 lymphocytic infiltration 179:202 lymphocytic infiltration 179:202 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 1 58 theme glands 246:251 arg1 hypofunction 208:219 hypofunction 208:219 hypofunction 208:219 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 7 59 theme Western 1391:1397 arg1 blotting 1399:1406 protein Western blotting 1383:1406 protein Western blotting 1383:1406 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 6 60 theme residual 1172:1179 arg1 saliva 1189:1194 residual mucosal saliva 1172:1194 residual mucosal saliva 1172:1194 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 8 61 theme core 1628:1631 arg1 sulfation 1635:1643 core 1 sulfation 1628:1643 core 1 sulfation 1628:1643 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 10 62 theme disialylated 1896:1907 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 2 63 theme salivary 267:274 arg1 function 276:283 salivary function 267:283 salivary function 267:283 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 1 64 theme autoimmune 142:151 arg1 disorder 153:160 a chronic autoimmune disorder 132:160 a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands 132:251 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 1 64 theme autoimmune 142:151 arg1 syndrome 120:127 Sjögren's syndrome 110:127 Sjögren's syndrome 110:127 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 10 65 from decrease 1851:1858 arg1 wetness 1945:1951 residual mucosal wetness 1928:1951 residual mucosal wetness 1928:1951 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 65 from decrease 1851:1858 arg1 MUC7 1922:1925 MUC7 1922:1925 MUC7 1922:1925 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 65 from decrease 1851:1858 arg1 rate 1982:1985 whole mouth saliva flow rate 1958:1985 whole mouth saliva flow rate 1958:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 65 from decrease 1851:1858 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 7 66 theme glycan 1412:1417 arg1 staining 1419:1426 glycan staining 1412:1426 glycan staining 1412:1426 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 10 67 theme oral 2058:2061 arg1 dryness 2063:2069 oral dryness 2058:2069 oral dryness 2058:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 4 68 theme major 639:643 arg1 mucins 690:695 heavily glycosylated mucins 669:695 heavily glycosylated mucins MUC5B and MUC7 669:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 68 theme major 639:643 arg1 constituents 645:656 major constituents 639:656 major constituents 639:656 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 10 69 theme core 1889:1892 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 5 70 from saliva 981:986 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 70 from saliva 981:986 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 7 71 from reduction 1441:1449 arg1 Sjögren 1477:1483 Sjögren 1477:1483 Sjögren 1477:1483 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 7 71 from reduction 1441:1449 arg1 glycosylation 1460:1472 mucin glycosylation 1454:1472 mucin glycosylation 1454:1472 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 11 72 theme observed 2076:2083 arg1 changes 2085:2091 The observed changes 2072:2091 The observed changes in MUC7 glycosylation 2072:2113 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 11 72 theme observed 2076:2083 arg1 tool 2147:2150 a potential diagnostic tool 2124:2150 a potential diagnostic tool for saliva quality 2124:2169 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 7 73 theme blotting 1399:1406 arg1 comparison 1369:1378 a comparison 1367:1378 a comparison of protein Western blotting and glycan staining 1367:1426 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 2 74 theme oral 294:297 arg1 dryness 299:305 oral dryness 294:305 oral dryness 294:305 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 1 75 theme lymphocytic 179:189 arg1 infiltration 191:202 lymphocytic infiltration 179:202 lymphocytic infiltration 179:202 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 7 76 theme protein 1383:1389 arg1 blotting 1399:1406 protein Western blotting 1383:1406 protein Western blotting 1383:1406 Although the concentration of mucins MUC5B and MUC7 were similar between patients and controls, a comparison of protein Western blotting and glycan staining identified a reduction in mucin glycosylation in Sjögren's, particularly on MUC7. 26631508 11 77 theme multifactorial 2230:2243 arg1 syndrome 2245:2252 this multifactorial syndrome 2225:2252 this multifactorial syndrome 2225:2252 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 4 78 theme complicated 605:615 arg1 fluid 628:632 a complicated biological fluid 603:632 a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7 603:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 78 theme complicated 605:615 arg1 Saliva 593:598 Saliva 593:598 Saliva 593:598 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 3 79 theme patient 571:577 arg1 presentation 579:590 patient presentation 571:590 patient presentation 571:590 Despite the implications of this prevalent disease, diagnosis currently takes years, partly due to the diversity in patient presentation. 26631508 5 80 from surface 1003:1009 arg1 perception 830:839 Sjögren's patient's perception 810:839 Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) 810:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 81 theme dryness 844:850 arg1 perception 830:839 Sjögren's patient's perception 810:839 Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) 810:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 11 82 theme potential 2126:2134 arg1 changes 2085:2091 The observed changes 2072:2091 The observed changes in MUC7 glycosylation 2072:2113 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 11 82 theme potential 2126:2134 arg1 tool 2147:2150 a potential diagnostic tool 2124:2150 a potential diagnostic tool for saliva quality 2124:2169 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 8 83 attach released 1541:1548 arg2 O-glycans 1531:1539 O-glycans 1531:1539 O-glycans released from MUC7 by β-elimination 1531:1575 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 8 83 attach released 1541:1548 arg1 MUC7 1555:1558 MUC7 1555:1558 MUC7 1555:1558 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 10 84 gly disialylated 1896:1907 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 85 theme mucosal 1937:1943 arg1 wetness 1945:1951 residual mucosal wetness 1928:1951 residual mucosal wetness 1928:1951 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 6 86 theme salivary 1197:1204 arg1 rate 1211:1214 salivary flow rate 1197:1214 salivary flow rate 1197:1214 Sjögren's patients exhibited a statistically significant reduction in residual mucosal saliva, salivary flow rate, and extensional rheology, spinnbarkeit (stringiness). 26631508 10 87 theme saliva 1970:1975 arg1 flow 1977:1980 whole mouth saliva flow 1958:1980 whole mouth saliva flow rate 1958:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 10 88 theme whole 1958:1962 arg1 flow 1977:1980 whole mouth saliva flow 1958:1980 whole mouth saliva flow rate 1958:1985 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 11 89 theme saliva 2156:2161 arg1 quality 2163:2169 saliva quality 2156:2169 saliva quality 2156:2169 The observed changes in MUC7 glycosylation could be a potential diagnostic tool for saliva quality and taken into consideration for future therapies for this multifactorial syndrome. 26631508 5 90 theme whole 954:958 arg1 saliva 966:971 whole mouth saliva 954:971 whole mouth saliva 954:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 8 91 theme LC-MS/MS 1510:1517 arg1 analysis 1519:1526 LC-MS/MS analysis 1510:1526 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination 1510:1575 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 5 92 theme mouth 960:964 arg1 saliva 966:971 whole mouth saliva 954:971 whole mouth saliva 954:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 4 93 theme hydrating 748:756 arg1 properties 774:783 its viscoelastic and hydrating and lubricating properties 727:783 its viscoelastic and hydrating and lubricating properties 727:783 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 0 94 theme Oral 96:99 arg1 Dryness 101:107 Oral Dryness 96:107 Oral Dryness 96:107 Reduced Mucin-7 (Muc7) Sialylation and Altered Saliva Rheology in Sjögren's Syndrome Associated Oral Dryness. 26631508 5 95 theme saliva 966:971 arg1 analysis 942:949 glycan analysis 935:949 glycan analysis of whole mouth saliva 935:971 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 8 96 contain had 1609:1611 arg1 patients 1600:1607 patients 1600:1607 patients 1600:1607 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 8 96 contain had 1609:1611 arg2 increase 1616:1623 an increase 1613:1623 an increase in core 1 sulfation 1613:1643 LC-MS/MS analysis of O-glycans released from MUC7 by β-elimination revealed that although patients had an increase in core 1 sulfation, the even larger reduction in sialylation resulted in a global decline of charged glycans. 26631508 4 97 theme lubricating 762:772 arg1 properties 774:783 its viscoelastic and hydrating and lubricating properties 727:783 its viscoelastic and hydrating and lubricating properties 727:783 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 0 98 theme Mucin-7 8:14 arg1 Sialylation 23:33 Reduced Mucin-7 (Muc7) Sialylation 0:33 Reduced Mucin-7 (Muc7) Sialylation 0:33 Reduced Mucin-7 (Muc7) Sialylation and Altered Saliva Rheology in Sjögren's Syndrome Associated Oral Dryness. 26631508 4 99 dep important 713:721 arg1 fluid 628:632 a complicated biological fluid 603:632 a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7 603:710 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 4 99 dep important 713:721 arg1 Saliva 593:598 Saliva 593:598 Saliva 593:598 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 10 100 from effect 2030:2035 arg1 perception 2044:2053 the perception 2040:2053 the perception of oral dryness 2040:2069 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 3 101 from diversity 558:566 arg1 presentation 579:590 patient presentation 571:590 patient presentation 571:590 Despite the implications of this prevalent disease, diagnosis currently takes years, partly due to the diversity in patient presentation. 26631508 9 102 theme core 1787:1790 arg1 structure 1807:1815 the extended core 2 disialylated structure 1774:1815 the extended core 2 disialylated structure 1774:1815 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 4 103 theme viscoelastic 731:742 arg1 properties 774:783 its viscoelastic and hydrating and lubricating properties 727:783 its viscoelastic and hydrating and lubricating properties 727:783 Saliva is a complicated biological fluid with major constituents, including heavily glycosylated mucins MUC5B and MUC7, important for its viscoelastic and hydrating and lubricating properties. 26631508 5 104 theme mucosal 995:1001 arg1 surface 1003:1009 the mucosal surface 991:1009 the mucosal surface (residual mucosal saliva) 991:1035 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 5 104 theme mucosal 995:1001 arg1 saliva 1029:1034 residual mucosal saliva 1012:1034 residual mucosal saliva 1012:1034 This study investigated Sjögren's patient's perception of dryness (bother index questionnaires) along with the rheological, protein composition, and glycan analysis of whole mouth saliva and the saliva on the mucosal surface (residual mucosal saliva) to understand the properties that most affect patient wellbeing. 26631508 9 105 theme disialylated 1794:1805 arg1 structure 1807:1815 the extended core 2 disialylated structure 1774:1815 the extended core 2 disialylated structure 1774:1815 This was primarily due to the loss of the extended core 2 disialylated structure, with and without fucosylation. 26631508 2 106 theme other 387:391 arg1 diseases 404:411 other autoimmune diseases 387:411 other autoimmune diseases 387:411 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 0 107 theme Altered 39:45 arg1 Rheology 54:61 Altered Saliva Rheology 39:61 Altered Saliva Rheology 39:61 Reduced Mucin-7 (Muc7) Sialylation and Altered Saliva Rheology in Sjögren's Syndrome Associated Oral Dryness. 26631508 2 108 theme certain 438:444 arg1 cancers 446:452 certain cancers 438:452 certain cancers 438:452 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 2 109 theme impaired 308:315 arg1 swallowing 317:326 impaired swallowing 308:326 impaired swallowing 308:326 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 1 110 theme salivary 224:231 arg1 glands 246:251 salivary and lacrimal glands 224:251 glands 246:251 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26631508 10 111 theme fucosylated 1877:1887 arg1 structure 1909:1917 the extended, fucosylated core 2 disialylated structure 1863:1917 structure 1909:1917 A decrease in the extended, fucosylated core 2 disialylated structure on MUC7, residual mucosal wetness, and whole mouth saliva flow rate appeared to have a negative and cumulative effect on the perception of oral dryness. 26631508 2 112 theme increased 344:352 arg1 infection 354:362 increased infection 344:362 increased infection 344:362 This loss of salivary function leads to oral dryness, impaired swallowing and speech, and increased infection and is associated with other autoimmune diseases and an increased risk of certain cancers. 26631508 1 113 theme lacrimal 237:244 arg1 glands 246:251 salivary and lacrimal glands 224:251 glands 246:251 Sjögren's syndrome is a chronic autoimmune disorder characterized by lymphocytic infiltration and hypofunction of salivary and lacrimal glands. 26171609 9 0 theme cytoplasmic 1527:1537 arg1 face 1539:1542 the cytoplasmic face 1523:1542 the cytoplasmic face of the plasma membrane of breast cancer cells 1523:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 6 1 theme cells 1018:1022 arg1 regions 988:994 the perinuclear regions 972:994 the perinuclear regions of mammary epithelial cells 972:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 3 2 theme extensive 610:618 arg1 branching 649:657 extensive terminal N-acetylglucosamine branching 610:657 extensive terminal N-acetylglucosamine branching 610:657 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 1 3 from regulation 288:297 arg1 track 346:350 human gastrointestinal track 323:350 human gastrointestinal track 323:350 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 9 4 theme plasma 1551:1556 arg1 membrane 1558:1565 the plasma membrane 1547:1565 the plasma membrane of breast cancer cells 1547:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 10 5 theme cell 1783:1786 arg1 surface 1788:1794 the cell surface 1779:1794 the cell surface 1779:1794 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 11 6 theme different 1863:1871 arg1 distribution 1885:1896 different subcellular distribution 1863:1896 different subcellular distribution 1863:1896 Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases. 26171609 3 7 with type 600:603 arg1 branching 649:657 extensive terminal N-acetylglucosamine branching 610:657 extensive terminal N-acetylglucosamine branching 610:657 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 1 8 theme transmembrane 160:172 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 8 theme transmembrane 160:172 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 9 theme Kunitz-type 175:185 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 9 theme Kunitz-type 175:185 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 6 10 theme immature 906:913 arg1 form 891:894 The 25-kDa form 880:894 The 25-kDa form 880:894 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 6 10 theme immature 906:913 arg1 HAI-2 915:919 likely immature HAI-2 899:919 likely immature HAI-2 899:919 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 9 11 theme cancer 1577:1582 arg1 cells 1584:1588 breast cancer cells 1570:1588 breast cancer cells 1570:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 7 12 theme 30-40-kDa 1170:1178 arg1 HAI-2 1180:1184 the 30-40-kDa HAI-2 1166:1184 the 30-40-kDa HAI-2 1166:1184 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 1 13 theme protease 194:201 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 13 theme protease 194:201 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 11 14 theme different 1925:1933 arg1 proteases 1942:1950 different target proteases 1925:1950 different target proteases 1925:1950 Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases. 26171609 3 15 theme oligomannose 537:548 arg1 type 550:553 the oligomannose type 533:553 the oligomannose type 533:553 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 5 16 theme 30-40-kDa 795:803 arg1 HAI-2 824:828 mature HAI-2 817:828 mature HAI-2 817:828 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 5 16 theme 30-40-kDa 795:803 arg1 form 805:808 The 30-40-kDa form 791:808 The 30-40-kDa form 791:808 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 6 17 theme 25-kDa 884:889 arg1 form 891:894 The 25-kDa form 880:894 The 25-kDa form 880:894 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 6 17 theme 25-kDa 884:889 arg1 HAI-2 915:919 likely immature HAI-2 899:919 likely immature HAI-2 899:919 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 10 18 theme membrane 1668:1675 arg1 face 1653:1656 the extracellular face 1635:1656 the extracellular face of plasma membrane 1635:1675 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 4 19 theme different 668:676 arg1 types 678:682 The two different types 660:682 The two different types of N-glycan 660:694 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 1 20 theme SPINT 136:140 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 20 theme SPINT 136:140 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 8 21 theme 30-40-kDa 1312:1320 arg1 HAI-2 1322:1326 the 30-40-kDa HAI-2 1308:1326 the 30-40-kDa HAI-2 1308:1326 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 8 22 theme cell 1377:1380 arg1 surface 1382:1388 the cell surface 1373:1388 the cell surface 1373:1388 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 7 23 theme cells 1253:1257 arg1 cells 1253:1257 cells 1253:1257 cells 1253:1257 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 7 23 theme cells 1253:1257 arg1 variety 1242:1248 a variety 1240:1248 a variety of cells 1240:1257 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 0 24 theme Inhibition 63:72 arg1 Distribution 43:54 the Subcellular Distribution 27:54 the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin 27:113 N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. 26171609 5 25 located localized 848:856 arg2 HAI-2 824:828 mature HAI-2 817:828 mature HAI-2 817:828 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 5 25 located localized 848:856 arg2 form 805:808 The 30-40-kDa form 791:808 The 30-40-kDa form 791:808 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 5 25 located localized 848:856 arg1 vesicles/granules 861:877 vesicles/granules 861:877 vesicles/granules 861:877 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 0 26 theme Matriptase 77:86 arg1 Inhibition 63:72 and Inhibition 59:72 Inhibition 63:72 N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. 26171609 1 27 theme human 323:327 arg1 track 346:350 human gastrointestinal track 323:350 human gastrointestinal track 323:350 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 2 28 dep size 421:424 arg1 25-kDa 441:446 25-kDa 441:446 25-kDa 441:446 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 2 28 dep size 421:424 arg1 30-40- 427:432 30-40- 427:432 30-40- 427:432 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 7 29 theme different 1033:1041 arg1 N-glycans 1043:1051 The two different N-glycans 1025:1051 The two different N-glycans 1025:1051 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 0 30 theme HAI-2/Placental 91:105 arg1 Bikunin 107:113 HAI-2/Placental Bikunin 91:113 HAI-2/Placental Bikunin 91:113 N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. 26171609 8 31 theme breast 1263:1268 arg1 cells 1277:1281 breast cancer cells 1263:1281 breast cancer cells 1263:1281 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 10 32 located detected 1623:1630 arg1 face 1653:1656 the extracellular face 1635:1656 the extracellular face of plasma membrane 1635:1675 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 10 32 located detected 1623:1630 arg2 HAI-2 1608:1612 the 25-kDa HAI-2 1597:1612 the 25-kDa HAI-2 1597:1612 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 10 32 located detected 1623:1630 arg1 levels 1689:1694 very low levels 1680:1694 very low levels 1680:1694 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 2 33 theme different 411:419 arg1 size 421:424 different size 411:424 different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57 411:484 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 8 34 theme matriptase-HAI-2 1419:1434 arg1 complex 1436:1442 the matriptase-HAI-2 complex 1415:1442 the matriptase-HAI-2 complex 1415:1442 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 0 35 theme N-Glycan 0:7 arg1 Branching 9:17 N-Glycan Branching 0:17 N-Glycan Branching 0:17 N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. 26171609 6 36 theme perinuclear 976:986 arg1 regions 988:994 the perinuclear regions 972:994 the perinuclear regions of mammary epithelial cells 972:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 7 37 theme maturation 1091:1100 arg1 stages 1102:1107 different maturation stages 1081:1107 different maturation stages of N-glycosylation 1081:1126 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 7 38 theme similar 1226:1232 arg1 levels 1211:1216 their levels 1205:1216 their levels roughly similar among a variety of cells 1205:1257 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 0 39 theme Subcellular 31:41 arg1 Distribution 43:54 the Subcellular Distribution 27:54 the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin 27:113 N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin. 26171609 7 40 theme levels 1211:1216 arg1 ratio 1196:1200 the ratio 1192:1200 the ratio of their levels roughly similar among a variety of cells 1192:1257 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 4 41 theme HAI-2 779:783 arg1 mAbs 785:788 two different HAI-2 mAbs 765:788 two different HAI-2 mAbs 765:788 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 1 42 theme absorption 309:318 arg1 regulation 288:297 regulation 288:297 regulation of sodium absorption in human gastrointestinal track 288:350 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 10 43 contain have 1710:1713 arg1 it 1696:1697 it 1696:1697 it 1696:1697 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 10 43 contain have 1710:1713 arg2 role 1718:1721 no role 1715:1721 no role 1715:1721 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 8 44 theme HAI-2 1322:1326 arg1 HAI-2 1322:1326 the 30-40-kDa HAI-2 1308:1326 the 30-40-kDa HAI-2 1308:1326 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 8 44 theme HAI-2 1322:1326 arg1 amount 1298:1303 a significant amount 1284:1303 a significant amount of the 30-40-kDa HAI-2 1284:1326 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 3 45 theme complex 592:598 arg1 type 600:603 complex type 592:603 complex type 592:603 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 10 46 theme 25-kDa 1601:1606 arg1 HAI-2 1608:1612 the 25-kDa HAI-2 1597:1612 the 25-kDa HAI-2 1597:1612 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 1 47 theme placenta 250:257 arg1 bikunin 259:265 placenta bikunin 250:265 placenta bikunin (PB) 250:270 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 47 theme placenta 250:257 arg1 PB 268:269 PB 268:269 PB 268:269 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 6 48 theme epithelial 1007:1016 arg1 cells 1018:1022 mammary epithelial cells 999:1022 mammary epithelial cells 999:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 3 49 theme terminal 620:627 arg1 branching 649:657 extensive terminal N-acetylglucosamine branching 610:657 extensive terminal N-acetylglucosamine branching 610:657 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 11 50 theme subcellular 1873:1883 arg1 distribution 1885:1896 different subcellular distribution 1863:1896 different subcellular distribution 1863:1896 Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases. 26171609 9 51 theme membrane 1558:1565 arg1 face 1539:1542 the cytoplasmic face 1523:1542 the cytoplasmic face of the plasma membrane of breast cancer cells 1523:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 7 52 theme a 1151:1151 arg1 precursor 1153:1161 the 25-kDa likely a precursor 1133:1161 the 25-kDa likely a precursor of the 30-40-kDa HAI-2 1133:1184 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 10 53 theme plasma 1661:1666 arg1 membrane 1668:1675 plasma membrane 1661:1675 plasma membrane 1661:1675 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 9 54 theme breast 1570:1575 arg1 cells 1584:1588 breast cancer cells 1570:1588 breast cancer cells 1570:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 3 55 theme 25-kDa 503:508 arg1 HAI-2 510:514 25-kDa HAI-2 503:514 25-kDa HAI-2 503:514 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 1 56 theme serine 187:192 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 56 theme serine 187:192 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 10 57 from paucity 1768:1774 arg1 surface 1788:1794 the cell surface 1779:1794 the cell surface 1779:1794 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 6 58 theme likely 899:904 arg1 form 891:894 The 25-kDa form 880:894 The 25-kDa form 880:894 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 6 58 theme likely 899:904 arg1 HAI-2 915:919 likely immature HAI-2 899:919 likely immature HAI-2 899:919 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 9 59 theme cells 1584:1588 arg1 membrane 1558:1565 the plasma membrane 1547:1565 the plasma membrane of breast cancer cells 1547:1588 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 10 60 theme low 1685:1687 arg1 levels 1689:1694 very low levels 1680:1694 very low levels 1680:1694 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 5 61 theme mature 817:822 arg1 HAI-2 824:828 mature HAI-2 817:828 mature HAI-2 817:828 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 5 61 theme mature 817:822 arg1 form 805:808 The 30-40-kDa form 791:808 The 30-40-kDa form 791:808 The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. 26171609 11 62 theme target 1935:1940 arg1 proteases 1942:1950 different target proteases 1925:1950 different target proteases 1925:1950 Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases. 26171609 1 63 theme gene 120:123 arg1 inhibitor 203:211 a transmembrane, Kunitz-type serine protease inhibitor 158:211 inhibitor 203:211 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 1 63 theme gene 120:123 arg1 product 125:131 The gene product 116:131 The gene product 116:131 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 2 64 from N-glycans 466:474 arg1 Asn-57 479:484 Asn-57 479:484 Asn-57 479:484 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 8 65 from matriptase 1359:1368 arg1 surface 1382:1388 the cell surface 1373:1388 the cell surface 1373:1388 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 11 66 theme N-glycan 1820:1827 arg1 branching 1829:1837 N-glycan branching 1820:1837 N-glycan branching 1820:1837 Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases. 26171609 6 67 theme endoplasmic 942:952 arg1 ER 965:966 ER 965:966 ER 965:966 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 6 67 theme endoplasmic 942:952 arg1 reticulum 954:962 the endoplasmic reticulum 938:962 the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells 938:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 2 68 theme due 449:451 arg1 size 421:424 different size 411:424 different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57 411:484 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 7 69 theme 25-kDa 1137:1142 arg1 precursor 1153:1161 the 25-kDa likely a precursor 1133:1161 the 25-kDa likely a precursor of the 30-40-kDa HAI-2 1133:1184 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 7 70 theme likely 1144:1149 arg1 precursor 1153:1161 the 25-kDa likely a precursor 1133:1161 the 25-kDa likely a precursor of the 30-40-kDa HAI-2 1133:1184 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 3 71 from N-glycan 491:498 arg1 HAI-2 510:514 25-kDa HAI-2 503:514 25-kDa HAI-2 503:514 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 1 72 theme gastrointestinal 329:344 arg1 track 346:350 human gastrointestinal track 323:350 human gastrointestinal track 323:350 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 4 73 from epitopes 720:727 arg1 polypeptide 738:748 HAI-2 polypeptide 732:748 HAI-2 polypeptide 732:748 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 10 74 theme extracellular 1639:1651 arg1 face 1653:1656 the extracellular face 1635:1656 the extracellular face of plasma membrane 1635:1675 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 6 75 from reticulum 954:962 arg1 regions 988:994 the perinuclear regions 972:994 the perinuclear regions of mammary epithelial cells 972:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 7 76 theme N-glycosylation 1112:1126 arg1 stages 1102:1107 different maturation stages 1081:1107 different maturation stages of N-glycosylation 1081:1126 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 9 77 theme 25-kDa 1449:1454 arg1 HAI-2 1456:1460 The 25-kDa HAI-2 1445:1460 The 25-kDa HAI-2 1445:1460 The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. 26171609 6 78 theme mammary 999:1005 arg1 cells 1018:1022 mammary epithelial cells 999:1022 mammary epithelial cells 999:1022 The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. 26171609 2 79 theme different 456:464 arg1 N-glycans 466:474 different N-glycans 456:474 different N-glycans on Asn-57 456:484 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 8 80 theme complex 1436:1442 arg1 shedding 1403:1410 shedding 1403:1410 shedding of the matriptase-HAI-2 complex 1403:1442 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 10 81 theme matriptase 1726:1735 arg1 inhibition 1737:1746 matriptase inhibition 1726:1746 matriptase inhibition 1726:1746 While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. 26171609 8 82 theme cancer 1270:1275 arg1 cells 1277:1281 breast cancer cells 1263:1281 breast cancer cells 1263:1281 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 7 83 theme different 1081:1089 arg1 stages 1102:1107 different maturation stages 1081:1107 different maturation stages of N-glycosylation 1081:1126 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 4 84 theme N-glycan 687:694 arg1 types 678:682 The two different types 660:682 The two different types of N-glycan 660:694 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 3 85 theme 30-40-kDa 567:575 arg1 HAI-2 577:581 30-40-kDa HAI-2 567:581 30-40-kDa HAI-2 567:581 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 26171609 8 86 theme significant 1286:1296 arg1 HAI-2 1322:1326 the 30-40-kDa HAI-2 1308:1326 the 30-40-kDa HAI-2 1308:1326 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 8 86 theme significant 1286:1296 arg1 amount 1298:1303 a significant amount 1284:1303 a significant amount of the 30-40-kDa HAI-2 1284:1326 In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. 26171609 2 87 theme size 421:424 arg1 SPINT 372:376 SPINT 2 372:378 SPINT 2 372:378 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 2 87 theme size 421:424 arg1 species 400:406 two species 396:406 two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57 396:484 Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. 26171609 4 88 theme different 769:777 arg1 mAbs 785:788 two different HAI-2 mAbs 765:788 two different HAI-2 mAbs 765:788 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 1 89 theme sodium 302:307 arg1 absorption 309:318 sodium absorption 302:318 sodium absorption 302:318 The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. 26171609 4 90 theme HAI-2 732:736 arg1 polypeptide 738:748 HAI-2 polypeptide 732:748 HAI-2 polypeptide 732:748 The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. 26171609 7 91 theme HAI-2 1180:1184 arg1 precursor 1153:1161 the 25-kDa likely a precursor 1133:1161 the 25-kDa likely a precursor of the 30-40-kDa HAI-2 1133:1184 The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. 26171609 3 92 theme N-acetylglucosamine 629:647 arg1 branching 649:657 extensive terminal N-acetylglucosamine branching 610:657 extensive terminal N-acetylglucosamine branching 610:657 The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. 25868116 5 0 theme protected 994:1002 arg1 ethylthioglycoside 1017:1034 the selectively protected disaccharide ethylthioglycoside 978:1034 the selectively protected disaccharide ethylthioglycoside in good overall yield 978:1056 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 1 1 theme 2,4,6-tri-O-benzoyl-3-O-benzyl 193:222 arg1 imidate 239:245 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate 193:245 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate 193:245 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 5 2 theme disaccharide 1004:1015 arg1 ethylthioglycoside 1017:1034 the selectively protected disaccharide ethylthioglycoside 978:1034 the selectively protected disaccharide ethylthioglycoside in good overall yield 978:1056 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 5 3 from ethylthioglycoside 1017:1034 arg1 yield 1052:1056 good overall yield 1039:1056 good overall yield 1039:1056 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 8 4 theme second 1571:1576 arg1 hydrogenolysis 1578:1591 second hydrogenolysis 1571:1591 second hydrogenolysis 1571:1591 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 7 5 theme terminal 1378:1385 arg1 ether 1398:1402 the terminal 3-O-benzyl ether 1374:1402 the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside 1374:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 1 6 theme glucopyranosyl 224:237 arg1 imidate 239:245 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate 193:245 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate 193:245 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 6 7 theme disaccharide 1141:1152 arg1 glycoside 1161:1169 the disaccharide tether glycoside 1137:1169 the disaccharide tether glycoside 1137:1169 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 0 8 theme cell 88:91 arg1 Dectin-1 103:110 Dectin-1 103:110 Dectin-1 103:110 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 0 8 theme cell 88:91 arg1 receptor 93:100 the dendritic cell receptor 74:100 the dendritic cell receptor 74:100 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 6 9 theme catalytic 1181:1189 arg1 removal 1191:1197 catalytic removal 1181:1197 catalytic removal of benzyl ether 1181:1213 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 3 10 from unreactive 659:668 arg1 reaction 687:694 glycosylation reaction 673:694 glycosylation reaction with the debenzylated di- and trisaccharide alcohols 673:747 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 8 11 theme antigen 1795:1801 arg1 processing 1803:1812 improved antigen processing 1786:1812 improved antigen processing by dendritic cells 1786:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 4 12 theme perbenzoylated 766:779 arg1 derivative 803:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 0 13 theme dendritic 78:86 arg1 Dectin-1 103:110 Dectin-1 103:110 Dectin-1 103:110 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 0 13 theme dendritic 78:86 arg1 receptor 93:100 the dendritic cell receptor 74:100 the dendritic cell receptor 74:100 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 8 14 theme Dectin-1 1675:1682 arg1 ligand 1684:1689 a Dectin-1 ligand 1673:1689 a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells 1673:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 3 15 theme debenzylated 705:716 arg1 di- 718:720 debenzylated di- 705:720 debenzylated di- 705:720 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 4 16 theme 3'-O-benzyl 754:764 arg1 derivative 803:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 6 17 from thioglycoside 1274:1286 arg1 reaction 1297:1304 a 2+2 reaction 1291:1304 a 2+2 reaction 1291:1304 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 3 18 theme terminated 611:620 arg1 products 636:643 the furanose terminated glycosylation products 598:643 the furanose terminated glycosylation products 598:643 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 8 19 theme residual 1605:1612 arg1 group 1621:1625 a residual benzyl group 1603:1625 a residual benzyl group 1603:1625 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 5 20 theme overall 1044:1050 arg1 yield 1052:1056 good overall yield 1039:1056 good overall yield 1039:1056 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 2 21 link linked 498:503 arg1 trisaccharide 505:517 the β1,3 linked trisaccharide 489:517 the β1,3 linked trisaccharide in excellent yield 489:536 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 3 22 with reaction 687:694 arg1 di- 718:720 debenzylated di- 705:720 debenzylated di- 705:720 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 3 22 with reaction 687:694 arg1 alcohols 740:747 trisaccharide alcohols 726:747 trisaccharide alcohols 726:747 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 6 23 gly glycosylated 1254:1265 arg1 alcohol 1242:1248 the resulting disaccharide alcohol 1215:1248 the resulting disaccharide alcohol 1215:1248 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 2 24 theme excellent 522:530 arg1 yield 532:536 excellent yield 522:536 excellent yield 522:536 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 8 25 theme glycoconjugate 1739:1752 arg1 vaccines 1754:1761 glycoconjugate vaccines 1739:1761 glycoconjugate vaccines 1739:1761 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 3 26 theme trisaccharide 726:738 arg1 alcohols 740:747 trisaccharide alcohols 726:747 trisaccharide alcohols 726:747 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 4 27 theme corresponding 870:882 arg1 derivative 926:935 6'-di-O-benzylidene derivative 906:935 6'-di-O-benzylidene derivative 906:935 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 4 27 theme corresponding 870:882 arg1 4,6:4 899:903 the corresponding perbenzoylated 4,6:4' 866:904 the corresponding perbenzoylated 4,6:4' 866:904 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 6 28 dep glycoside 1161:1169 arg1 tether 1154:1159 tether 1154:1159 tether 1154:1159 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 6 29 theme 2+2 1293:1295 arg1 reaction 1297:1304 a 2+2 reaction 1291:1304 a 2+2 reaction 1291:1304 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 7 30 theme selective 1348:1356 arg1 deprotection 1358:1369 selective deprotection 1348:1369 selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside 1348:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 7 31 theme 3-O-benzyl 1387:1396 arg1 ether 1398:1402 the terminal 3-O-benzyl ether 1374:1402 the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside 1374:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 2 32 from di-O-isopropylidene-α-D-glucofuranose 351:387 arg1 glycosylation 407:419 glycosylation 407:419 glycosylation 407:419 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 2 32 from di-O-isopropylidene-α-D-glucofuranose 351:387 arg1 yield 397:401 high yield 392:401 high yield 392:401 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 2 33 gly glycosylated 338:349 arg1 di-O-isopropylidene-α-D-glucofuranose 351:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose 325:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether 325:478 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 8 34 theme hexasaccharide 1643:1656 arg1 glycoside 1658:1666 the target hexasaccharide glycoside 1 1632:1668 the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells 1632:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 7 35 theme deprotection 1358:1369 arg1 Repetition 1334:1343 Repetition 1334:1343 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside 1334:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 8 36 theme hexasaccharide 1520:1533 arg1 Hydrogenolysis 1497:1510 Hydrogenolysis 1497:1510 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group 1497:1625 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 2 37 theme disaccharide 428:439 arg1 glycosylation 407:419 glycosylation 407:419 glycosylation 407:419 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 2 37 theme disaccharide 428:439 arg1 yield 397:401 high yield 392:401 high yield 392:401 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 7 38 theme ether 1398:1402 arg1 deprotection 1358:1369 selective deprotection 1348:1369 selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside 1348:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 1 39 theme crystalline 273:283 arg1 derivatives 312:322 crystalline benzyl and per-benzoylated derivatives 273:322 crystalline benzyl and per-benzoylated derivatives 273:322 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 6 40 with Glycosidation 1059:1071 arg1 5-methoxycarbonylpentanol 1106:1130 5-methoxycarbonylpentanol 1106:1130 5-methoxycarbonylpentanol 1106:1130 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 3 41 dep di- 718:720 arg1 the 701:703 the 701:703 the 701:703 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 5 42 theme Lewis 938:942 arg1 glycosidation 959:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 1 43 theme benzyl 285:290 arg1 derivatives 312:322 crystalline benzyl and per-benzoylated derivatives 273:322 crystalline benzyl and per-benzoylated derivatives 273:322 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 0 44 theme hexasaccharide 28:41 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of a 1,3 β-glucan hexasaccharide 0:41 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 4 45 theme 6'-di-O-benzylidene 906:924 arg1 derivative 926:935 6'-di-O-benzylidene derivative 906:935 6'-di-O-benzylidene derivative 906:935 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 4 45 theme 6'-di-O-benzylidene 906:924 arg1 4,6:4 899:903 the corresponding perbenzoylated 4,6:4' 866:904 the corresponding perbenzoylated 4,6:4' 866:904 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 2 46 theme imidate 330:336 arg1 di-O-isopropylidene-α-D-glucofuranose 351:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose 325:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether 325:478 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 0 47 theme β-glucan 19:26 arg1 hexasaccharide 28:41 a 1,3 β-glucan hexasaccharide 13:41 a 1,3 β-glucan hexasaccharide 13:41 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 6 48 theme disaccharide 1229:1240 arg1 alcohol 1242:1248 the resulting disaccharide alcohol 1215:1248 the resulting disaccharide alcohol 1215:1248 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 3 49 theme trisaccharides 547:560 arg1 imidates 562:569 Di- and trisaccharides imidates 539:569 Di- and trisaccharides imidates 539:569 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 0 50 theme 1,3 15:17 arg1 hexasaccharide 28:41 a 1,3 β-glucan hexasaccharide 13:41 a 1,3 β-glucan hexasaccharide 13:41 Synthesis of a 1,3 β-glucan hexasaccharide designed to target vaccines to the dendritic cell receptor, Dectin-1. 25868116 8 51 theme dendritic 1817:1825 arg1 cells 1827:1831 dendritic cells 1817:1831 dendritic cells 1817:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 1 52 theme per-benzoylated 296:310 arg1 derivatives 312:322 crystalline benzyl and per-benzoylated derivatives 273:322 crystalline benzyl and per-benzoylated derivatives 273:322 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 5 53 theme good 1039:1042 arg1 yield 1052:1056 good overall yield 1039:1056 good overall yield 1039:1056 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 2 54 gly glycosylation 407:419 arg1 disaccharide 428:439 the disaccharide 424:439 the disaccharide 424:439 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 8 55 theme covalent 1716:1723 arg1 attachment 1725:1734 covalent attachment 1716:1734 covalent attachment to glycoconjugate vaccines 1716:1761 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 6 56 theme donor 1095:1099 arg1 Glycosidation 1059:1071 Glycosidation 1059:1071 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol 1059:1130 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 2 57 from trisaccharide 505:517 arg1 yield 532:536 excellent yield 522:536 excellent yield 522:536 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 2 58 theme ether 474:478 arg1 removal 447:453 removal 447:453 removal of the 3'-O-benzyl ether 447:478 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 3 59 theme Di- 539:541 arg1 imidates 562:569 Di- and trisaccharides imidates 539:569 Di- and trisaccharides imidates 539:569 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 5 60 theme acid 944:947 arg1 glycosidation 959:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25868116 7 61 theme protected 1471:1479 arg1 hexasaccharide 1481:1494 a protected hexasaccharide 1469:1494 a protected hexasaccharide 1469:1494 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 2 62 theme 3'-O-benzyl 462:472 arg1 ether 474:478 the 3'-O-benzyl ether 458:478 the 3'-O-benzyl ether 458:478 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 8 63 theme improved 1786:1793 arg1 processing 1803:1812 improved antigen processing 1786:1812 improved antigen processing by dendritic cells 1786:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 6 64 theme ether 1209:1213 arg1 removal 1191:1197 catalytic removal 1181:1197 catalytic removal of benzyl ether 1181:1213 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 7 65 theme disaccharide 1437:1448 arg1 thioglycoside 1450:1462 the disaccharide thioglycoside 1433:1462 the disaccharide thioglycoside 1433:1462 Repetition of selective deprotection of the terminal 3-O-benzyl ether followed by glycosylation by the disaccharide thioglycoside gave a protected hexasaccharide. 25868116 1 66 theme 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose 131:186 arg1 Transformation 113:126 Transformation 113:126 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate 113:245 Transformation of 3-O-benzyl-1,2:5,6-di-O-isopropylidene-α-D-glucofuranose into 2,4,6-tri-O-benzoyl-3-O-benzyl glucopyranosyl imidate proceeded efficiently via crystalline benzyl and per-benzoylated derivatives. 25868116 2 67 theme high 392:395 arg1 yield 397:401 high yield 392:401 high yield 392:401 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 4 68 theme perbenzoylated 884:897 arg1 derivative 926:935 6'-di-O-benzylidene derivative 906:935 6'-di-O-benzylidene derivative 906:935 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 4 68 theme perbenzoylated 884:897 arg1 4,6:4 899:903 the corresponding perbenzoylated 4,6:4' 866:904 the corresponding perbenzoylated 4,6:4' 866:904 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 3 69 theme glycosylation 622:634 arg1 products 636:643 the furanose terminated glycosylation products 598:643 the furanose terminated glycosylation products 598:643 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 4 70 theme pyranose 794:801 arg1 derivative 803:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 6 71 theme resulting 1219:1227 arg1 alcohol 1242:1248 the resulting disaccharide alcohol 1215:1248 the resulting disaccharide alcohol 1215:1248 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 6 72 theme thioglycoside 1081:1093 arg1 donor 1095:1099 this thioglycoside donor 1076:1099 this thioglycoside donor 1076:1099 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 3 73 theme glycosylation 673:685 arg1 reaction 687:694 glycosylation reaction 673:694 glycosylation reaction with the debenzylated di- and trisaccharide alcohols 673:747 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 4 74 theme disaccharide 781:792 arg1 derivative 803:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative 750:812 The 3'-O-benzyl perbenzoylated disaccharide pyranose derivative could be selectively debenzoylated and converted to the corresponding perbenzoylated 4,6:4',6'-di-O-benzylidene derivative. 25868116 8 75 theme benzyl 1614:1619 arg1 group 1621:1625 a residual benzyl group 1603:1625 a residual benzyl group 1603:1625 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 3 76 theme furanose 602:609 arg1 products 636:643 the furanose terminated glycosylation products 598:643 the furanose terminated glycosylation products 598:643 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 2 77 theme linked 498:503 arg1 trisaccharide 505:517 the β1,3 linked trisaccharide 489:517 the β1,3 linked trisaccharide in excellent yield 489:536 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 8 78 theme target 1636:1641 arg1 glycoside 1658:1666 the target hexasaccharide glycoside 1 1632:1668 the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells 1632:1831 Hydrogenolysis of this hexasaccharide followed by transesterification and second hydrogenolysis to remove a residual benzyl group gave the target hexasaccharide glycoside 1 as a Dectin-1 ligand functionalized to permit covalent attachment to glycoconjugate vaccines and thereby facilitate improved antigen processing by dendritic cells. 25868116 6 79 theme benzyl 1202:1207 arg1 ether 1209:1213 benzyl ether 1202:1213 benzyl ether 1202:1213 Glycosidation of this thioglycoside donor with 5-methoxycarbonylpentanol gave the disaccharide tether glycoside and after catalytic removal of benzyl ether the resulting disaccharide alcohol was glycosylated by the thioglycoside in a 2+2 reaction to yield a tetrasaccharide. 25868116 2 80 theme glycosylated 338:349 arg1 di-O-isopropylidene-α-D-glucofuranose 351:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose 325:387 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether 325:478 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 2 81 theme β1,3 493:496 arg1 trisaccharide 505:517 the β1,3 linked trisaccharide 489:517 the β1,3 linked trisaccharide in excellent yield 489:536 This imidate glycosylated di-O-isopropylidene-α-D-glucofuranose in high yield and glycosylation of the disaccharide after removal of the 3'-O-benzyl ether afforded the β1,3 linked trisaccharide in excellent yield. 25868116 3 82 from reaction 687:694 arg1 unreactive 659:668 unreactive 659:668 unreactive 659:668 Di- and trisaccharides imidates were readily prepared from the furanose terminated glycosylation products but both were unreactive in glycosylation reaction with the debenzylated di- and trisaccharide alcohols. 25868116 5 83 theme catalyzed 949:957 arg1 glycosidation 959:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation 938:971 Lewis acid catalyzed glycosidation gave the selectively protected disaccharide ethylthioglycoside in good overall yield. 25983113 7 0 from populations 1275:1285 arg1 coats 1264:1268 jelly coats 1258:1268 jelly coats from populations not adapted to acidity 1258:1308 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 10 1 theme maternal 1852:1859 arg1 effects 1861:1867 adaptive maternal effects 1843:1867 adaptive maternal effects 1843:1867 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 2 2 theme populations 345:355 arg1 responses 324:332 The ecological and evolutionary responses 292:332 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages 292:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 1 3 theme powerful 237:244 arg1 stress 144:149 Environmental stress 130:149 Environmental stress 130:149 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 1 3 theme powerful 237:244 arg1 force 259:263 a powerful evolutionary force 235:263 a powerful evolutionary force at ecological time scales 235:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 7 4 theme jelly 1258:1262 arg1 coats 1264:1268 jelly coats 1258:1268 jelly coats from populations not adapted to acidity 1258:1308 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 1 5 from scales 284:289 arg1 stress 144:149 Environmental stress 130:149 Environmental stress 130:149 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 1 5 from scales 284:289 arg1 force 259:263 a powerful evolutionary force 235:263 a powerful evolutionary force at ecological time scales 235:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 0 6 theme Mechanistic 0:10 arg1 basis 12:16 Mechanistic basis 0:16 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 9 7 theme intra-specific 1613:1626 arg1 variation 1628:1636 extensive intra-specific variation 1603:1636 extensive intra-specific variation 1603:1636 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 2 8 theme ecological 296:305 arg1 responses 324:332 The ecological and evolutionary responses 292:332 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages 292:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 7 9 from population 1222:1231 arg1 coats 1195:1199 jelly coats 1189:1199 jelly coats from an acid-adapted population 1189:1231 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 8 10 from pH 1359:1360 arg1 survival 1347:1354 survival 1347:1354 survival at pH 4.0 1347:1364 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 0 11 theme adaptive 21:28 arg1 effects 39:45 adaptive maternal effects 21:45 adaptive maternal effects 21:45 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 1 12 theme natural 204:210 arg1 populations 212:222 natural populations 204:222 natural populations 204:222 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 4 13 theme moor 699:702 arg1 arvalis 714:720 the moor frog Rana arvalis 695:720 the moor frog Rana arvalis 695:720 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 9 14 theme strong 1672:1677 arg1 selection 1679:1687 strong selection 1672:1687 strong selection on water balance and glycosylation status of egg jelly coats 1672:1748 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 4 15 theme Rana 709:712 arg1 arvalis 714:720 the moor frog Rana arvalis 695:720 the moor frog Rana arvalis 695:720 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 9 16 from selection 1679:1687 arg1 balance 1698:1704 water balance 1692:1704 water balance 1692:1704 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 9 16 from selection 1679:1687 arg1 status 1724:1729 glycosylation status 1710:1729 glycosylation status 1710:1729 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 8 17 theme charge 1403:1408 arg1 state 1410:1414 charge state 1403:1414 charge state 1403:1414 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 3 18 theme maternal 469:476 arg1 effects 478:484 maternal effects 469:484 maternal effects 469:484 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 4 19 theme Previous 675:682 arg1 studies 684:690 Previous studies 675:690 Previous studies on the moor frog Rana arvalis 675:720 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 0 20 theme jelly 52:56 arg1 balance 64:70 egg jelly water balance 48:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 7 21 theme jelly 1168:1172 arg1 coat 1174:1177 the egg jelly coat 1160:1177 the egg jelly coat 1160:1177 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 5 22 theme maternal 899:906 arg1 effects 908:914 these adaptive maternal effects 884:914 these adaptive maternal effects 884:914 However, the exact mechanisms underlying these adaptive maternal effects remain unknown. 25983113 6 23 theme charge 984:989 arg1 state 991:995 charge state 984:995 charge state 984:995 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 9 24 theme glycosylation 1710:1722 arg1 status 1724:1729 glycosylation status 1710:1729 glycosylation status 1710:1729 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 8 25 theme jelly 1423:1427 arg1 loss 1394:1397 water loss 1388:1397 water loss 1388:1397 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 8 25 theme jelly 1423:1427 arg1 state 1410:1414 charge state 1403:1414 charge state 1403:1414 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 0 26 theme embryonic 81:89 arg1 adaptation 91:100 embryonic adaptation 81:100 embryonic adaptation to acidity in Rana arvalis 81:127 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 6 27 theme balance 972:978 arg1 role 958:961 the role 954:961 the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis 954:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 4 28 theme adaptive 755:762 arg1 divergence 764:773 adaptive divergence 755:773 adaptive divergence 755:773 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 9 29 theme jelly 1738:1742 arg1 coats 1744:1748 egg jelly coats 1734:1748 egg jelly coats 1734:1748 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 2 30 theme early 384:388 arg1 life-stages 390:400 early life-stages 384:400 early life-stages 384:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 7 31 theme severe 1139:1144 arg1 loss 1152:1155 severe water loss 1139:1155 severe water loss 1139:1155 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 6 32 from role 958:961 arg1 adaptation 1046:1055 embryonic adaptation 1036:1055 embryonic adaptation to acid stress in three populations of R. arvalis 1036:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 8 33 theme embryonic 1321:1329 arg1 tolerance 1336:1344 embryonic acid tolerance 1321:1344 embryonic acid tolerance (survival at pH 4.0) 1321:1365 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 2 34 theme maternal 425:432 arg1 effects 434:440 maternal effects 425:440 maternal effects 425:440 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 1 35 theme time 279:282 arg1 scales 284:289 ecological time scales 268:289 ecological time scales 268:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 6 36 theme jelly 1021:1025 arg1 coats 1027:1031 egg jelly coats 1017:1031 egg jelly coats 1017:1031 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 8 37 theme jelly 1483:1487 arg1 balance 1495:1501 jelly water balance 1483:1501 jelly water balance 1483:1501 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 0 38 theme maternal 30:37 arg1 effects 39:45 adaptive maternal effects 21:45 adaptive maternal effects 21:45 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 7 39 theme more 1242:1245 arg1 water 1247:1251 more water 1242:1251 more water 1242:1251 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 4 40 theme acid 820:823 arg1 tolerance 832:840 embryonic acid stress tolerance 810:840 embryonic acid stress tolerance 810:840 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 2 41 theme natural 337:343 arg1 populations 345:355 natural populations 337:355 natural populations 337:355 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 9 42 theme extensive 1603:1611 arg1 variation 1628:1636 extensive intra-specific variation 1603:1636 extensive intra-specific variation 1603:1636 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 6 43 theme R. 1096:1097 arg1 arvalis 1099:1105 R. arvalis 1096:1105 R. arvalis 1096:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 10 44 theme tolerance 1829:1837 arg1 mechanisms 1794:1803 the molecular mechanisms 1780:1803 the molecular mechanisms of environmental stress tolerance and adaptive maternal effects 1780:1867 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 1 45 theme populations 212:222 arg1 persistence 189:199 persistence 189:199 persistence of natural populations 189:222 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 8 46 theme embryonic 1521:1529 arg1 adaptation 1531:1540 embryonic adaptation 1521:1540 embryonic adaptation 1521:1540 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 10 47 theme environmental 1808:1820 arg1 tolerance 1829:1837 environmental stress tolerance 1808:1837 environmental stress tolerance 1808:1837 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 4 48 theme acidification 784:796 arg1 gradient 798:805 an acidification gradient 781:805 an acidification gradient in embryonic acid stress tolerance 781:840 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 10 49 theme effects 1861:1867 arg1 mechanisms 1794:1803 the molecular mechanisms 1780:1803 the molecular mechanisms of environmental stress tolerance and adaptive maternal effects 1780:1867 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 4 50 from studies 684:690 arg1 arvalis 714:720 the moor frog Rana arvalis 695:720 the moor frog Rana arvalis 695:720 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 9 51 theme egg 1582:1584 arg1 coats 1586:1590 egg coats 1582:1590 egg coats 1582:1590 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 6 52 theme acid 1060:1063 arg1 stress 1065:1070 acid stress 1060:1070 acid stress 1060:1070 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 10 53 theme adaptive 1843:1850 arg1 effects 1861:1867 adaptive maternal effects 1843:1867 adaptive maternal effects 1843:1867 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 1 54 theme evolutionary 246:257 arg1 stress 144:149 Environmental stress 130:149 Environmental stress 130:149 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 1 54 theme evolutionary 246:257 arg1 force 259:263 a powerful evolutionary force 235:263 a powerful evolutionary force at ecological time scales 235:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 8 55 dep tolerance 1336:1344 arg1 survival 1347:1354 survival 1347:1354 survival at pH 4.0 1347:1364 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 6 56 theme arvalis 1099:1105 arg1 populations 1081:1091 three populations 1075:1091 three populations of R. arvalis 1075:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 6 57 from adaptation 1046:1055 arg1 populations 1081:1091 three populations 1075:1091 three populations of R. arvalis 1075:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 4 58 theme stress 825:830 arg1 tolerance 832:840 embryonic acid stress tolerance 810:840 embryonic acid stress tolerance 810:840 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 5 59 theme exact 856:860 arg1 mechanisms 862:871 the exact mechanisms 852:871 the exact mechanisms underlying these adaptive maternal effects 852:914 However, the exact mechanisms underlying these adaptive maternal effects remain unknown. 25983113 6 60 theme water 966:970 arg1 balance 972:978 water balance 966:978 water balance 966:978 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 6 60 theme water 966:970 arg1 potential 1003:1011 zeta potential 998:1011 zeta potential 998:1011 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 0 61 theme Rana 116:119 arg1 arvalis 121:127 Rana arvalis 116:127 Rana arvalis 116:127 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 3 62 theme egg 587:589 arg1 coats 591:595 egg coats 587:595 egg coats 587:595 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 8 63 theme water 1388:1392 arg1 loss 1394:1397 water loss 1388:1397 water loss 1388:1397 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 4 64 from gradient 798:805 arg1 tolerance 832:840 embryonic acid stress tolerance 810:840 embryonic acid stress tolerance 810:840 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 2 65 from life-stages 390:400 arg1 stress 374:379 environmental stress 360:379 environmental stress at early life-stages 360:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 0 66 from adaptation 91:100 arg1 arvalis 121:127 Rana arvalis 116:127 Rana arvalis 116:127 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 4 67 theme frog 704:707 arg1 arvalis 714:720 the moor frog Rana arvalis 695:720 the moor frog Rana arvalis 695:720 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 0 68 theme water 58:62 arg1 balance 64:70 egg jelly water balance 48:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 1 69 theme Environmental 130:142 arg1 stress 144:149 Environmental stress 130:149 Environmental stress 130:149 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 1 69 theme Environmental 130:142 arg1 force 259:263 a powerful evolutionary force 235:263 a powerful evolutionary force at ecological time scales 235:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 6 70 theme zeta 998:1001 arg1 balance 972:978 water balance 966:978 water balance 966:978 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 6 70 theme zeta 998:1001 arg1 potential 1003:1011 zeta potential 998:1011 zeta potential 998:1011 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 9 71 theme water 1692:1696 arg1 balance 1698:1704 water balance 1692:1704 water balance 1692:1704 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 7 72 theme egg 1164:1166 arg1 coat 1174:1177 the egg jelly coat 1160:1177 the egg jelly coat 1160:1177 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 5 73 theme adaptive 890:897 arg1 effects 908:914 these adaptive maternal effects 884:914 these adaptive maternal effects 884:914 However, the exact mechanisms underlying these adaptive maternal effects remain unknown. 25983113 6 74 theme state 991:995 arg1 role 958:961 the role 954:961 the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis 954:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 10 75 theme stress 1822:1827 arg1 tolerance 1829:1837 environmental stress tolerance 1808:1837 environmental stress tolerance 1808:1837 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 4 76 theme egg 737:739 arg1 coat 741:744 the egg coat 733:744 the egg coat 733:744 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 7 77 theme water 1146:1150 arg1 loss 1152:1155 severe water loss 1139:1155 severe water loss 1139:1155 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 9 78 theme egg 1734:1736 arg1 coats 1744:1748 egg jelly coats 1734:1748 egg jelly coats 1734:1748 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 8 79 theme charged 1457:1463 arg1 glycans 1465:1471 negatively charged glycans 1446:1471 negatively charged glycans 1446:1471 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 3 80 theme early 450:454 arg1 life-stages 456:466 early life-stages 450:466 early life-stages 450:466 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 3 81 dep coats 510:514 arg1 structures 535:544 the extracellular structures 517:544 the extracellular structures surrounding the embryo 517:567 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 6 82 theme embryonic 1036:1044 arg1 adaptation 1046:1055 embryonic adaptation 1036:1055 embryonic adaptation to acid stress in three populations of R. arvalis 1036:1105 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 9 83 theme coats 1744:1748 arg1 selection 1679:1687 strong selection 1672:1687 strong selection on water balance and glycosylation status of egg jelly coats 1672:1748 These results indicate that egg coats can harbor extensive intra-specific variation, probably facilitated in part via strong selection on water balance and glycosylation status of egg jelly coats. 25983113 7 84 theme acidic 1122:1127 arg1 pH 1129:1130 acidic pH 1122:1130 acidic pH 1122:1130 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 1 85 theme ecological 268:277 arg1 scales 284:289 ecological time scales 268:289 ecological time scales 268:289 Environmental stress, such as acidification, can challenge persistence of natural populations and act as a powerful evolutionary force at ecological time scales. 25983113 6 86 theme coats 1027:1031 arg1 balance 972:978 water balance 966:978 water balance 966:978 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 6 86 theme coats 1027:1031 arg1 state 991:995 charge state 984:995 charge state 984:995 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 6 86 theme coats 1027:1031 arg1 potential 1003:1011 zeta potential 998:1011 zeta potential 998:1011 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 8 87 theme water 1489:1493 arg1 balance 1495:1501 jelly water balance 1483:1501 jelly water balance 1483:1501 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 0 88 theme effects 39:45 arg1 basis 12:16 Mechanistic basis 0:16 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 6 89 theme egg 1017:1019 arg1 coats 1027:1031 egg jelly coats 1017:1031 egg jelly coats 1017:1031 Here, we investigated the role of water balance and charge state (zeta potential) of egg jelly coats in embryonic adaptation to acid stress in three populations of R. arvalis. 25983113 10 90 theme molecular 1784:1792 arg1 mechanisms 1794:1803 the molecular mechanisms 1780:1803 the molecular mechanisms of environmental stress tolerance and adaptive maternal effects 1780:1867 These findings shed light on the molecular mechanisms of environmental stress tolerance and adaptive maternal effects. 25983113 8 91 theme acid 1331:1334 arg1 tolerance 1336:1344 embryonic acid tolerance 1321:1344 embryonic acid tolerance (survival at pH 4.0) 1321:1365 Moreover, embryonic acid tolerance (survival at pH 4.0) correlated with both water loss and charge state of the jelly, indicating that negatively charged glycans influence jelly water balance and contribute to embryonic adaptation to acidity. 25983113 0 92 theme egg 48:50 arg1 balance 64:70 egg jelly water balance 48:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 7 93 theme acid-adapted 1209:1220 arg1 population 1222:1231 an acid-adapted population 1206:1231 an acid-adapted population 1206:1231 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 2 94 theme environmental 360:372 arg1 stress 374:379 environmental stress 360:379 environmental stress at early life-stages 360:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 3 95 theme environmental 653:665 arg1 stress 667:672 environmental stress 653:672 environmental stress 653:672 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 4 96 theme embryonic 810:818 arg1 tolerance 832:840 embryonic acid stress tolerance 810:840 embryonic acid stress tolerance 810:840 Previous studies on the moor frog Rana arvalis found that the egg coat mediated adaptive divergence along an acidification gradient in embryonic acid stress tolerance. 25983113 3 97 theme egg 506:508 arg1 coats 510:514 egg coats 506:514 egg coats (the extracellular structures surrounding the embryo) 506:568 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 3 98 theme coats 591:595 arg1 role 579:582 the role 575:582 the role of egg coats 575:595 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 3 99 theme adaptation 639:648 arg1 context 628:634 the context 624:634 the context of adaptation to environmental stress 624:672 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25983113 2 100 theme evolutionary 311:322 arg1 responses 324:332 The ecological and evolutionary responses 292:332 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages 292:400 The ecological and evolutionary responses of natural populations to environmental stress at early life-stages are often mediated via maternal effects. 25983113 0 101 dep basis 12:16 arg1 balance 64:70 egg jelly water balance 48:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance 0:70 Mechanistic basis of adaptive maternal effects: egg jelly water balance mediates embryonic adaptation to acidity in Rana arvalis. 25983113 7 102 theme jelly 1189:1193 arg1 coats 1195:1199 jelly coats 1189:1199 jelly coats from an acid-adapted population 1189:1231 We found that acidic pH causes severe water loss in the egg jelly coat, but that jelly coats from an acid-adapted population retained more water than jelly coats from populations not adapted to acidity. 25983113 3 103 theme extracellular 521:533 arg1 structures 535:544 the extracellular structures 517:544 the extracellular structures surrounding the embryo 517:567 During early life-stages, maternal effects commonly arise from egg coats (the extracellular structures surrounding the embryo), but the role of egg coats has rarely been studied in the context of adaptation to environmental stress. 25674920 0 0 theme Streptococcus 60:72 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 3 1 theme target 618:623 arg1 molecule 625:632 The target molecule 614:632 The target molecule 614:632 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 2 2 theme main-chain 519:528 arg1 end 539:541 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 0 3 theme B 58:58 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 1 4 with pentasaccharide 274:288 arg1 branch 297:302 a branch 295:302 a branch 295:302 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 3 5 theme branched 744:751 arg1 acceptor 770:777 a branched trisaccharide as acceptor 742:777 a branched trisaccharide as acceptor 742:777 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 3 5 theme branched 744:751 arg1 sialo-disaccharide 710:727 a sialo-disaccharide 708:727 a sialo-disaccharide 708:727 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 0 6 theme polysaccharide 83:96 arg1 unit 36:39 the repeating unit 22:39 the repeating unit of type Ia group B Streptococcus capsular polysaccharide 22:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 3 7 theme glycosylation 675:687 arg1 strategy 689:696 3] glycosylation strategy 672:696 3] glycosylation strategy 672:696 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 0 8 theme capsular 74:81 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 2 9 theme Ia 443:444 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 10 theme further 553:559 arg1 elaboration 561:571 further elaboration 553:571 further elaboration 553:571 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 11 theme serotype 434:441 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 12 theme difficult 310:318 arg1 linkage 334:340 a difficult α-sialic acid linkage 308:340 a difficult α-sialic acid linkage 308:340 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 2 13 theme carrier 596:602 arg1 proteins 604:611 carrier proteins 596:611 carrier proteins 596:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 14 theme serotype 153:160 arg1 GBS 188:190 serotype Ia group B Streptococcus (GBS) 153:191 serotype Ia group B Streptococcus (GBS) 153:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 2 15 theme effective 392:400 arg1 synthesis 402:410 an effective synthesis 389:410 an effective synthesis 389:410 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 4 16 theme unit 858:861 arg1 oligomers 831:839 oligomers 831:839 oligomers of the repeating unit 831:861 The strategy was designed to suit the synthesis of oligomers of the repeating unit. 25674920 1 17 theme Ia 162:163 arg1 GBS 188:190 serotype Ia group B Streptococcus (GBS) 153:191 serotype Ia group B Streptococcus (GBS) 153:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 0 18 theme Chemical 0:7 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 0:97 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 2 19 theme reducing 530:537 arg1 end 539:541 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 20 contain had 476:478 arg2 functionality 491:503 a reactive functionality 480:503 a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 480:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 20 contain had 476:478 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 21 theme group 165:169 arg1 GBS 188:190 serotype Ia group B Streptococcus (GBS) 153:191 serotype Ia group B Streptococcus (GBS) 153:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 22 theme repeating 235:243 arg1 unit 245:248 its repeating unit 231:248 its repeating unit 231:248 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 22 theme repeating 235:243 arg1 pentasaccharide 274:288 a challenging pentasaccharide 260:288 a challenging pentasaccharide with a branch 260:302 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 4 23 theme oligomers 831:839 arg1 synthesis 818:826 the synthesis 814:826 the synthesis of oligomers of the repeating unit 814:861 The strategy was designed to suit the synthesis of oligomers of the repeating unit. 25674920 1 24 theme B 171:171 arg1 GBS 188:190 serotype Ia group B Streptococcus (GBS) 153:191 serotype Ia group B Streptococcus (GBS) 153:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 25 theme acid 329:332 arg1 linkage 334:340 a difficult α-sialic acid linkage 308:340 a difficult α-sialic acid linkage 308:340 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 26 theme Streptococcus 173:185 arg1 GBS 188:190 serotype Ia group B Streptococcus (GBS) 153:191 serotype Ia group B Streptococcus (GBS) 153:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 0 27 theme unit 36:39 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 0:97 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 3 28 theme convergent 656:665 arg1 [2 667:668 a convergent [2 654:668 a convergent [2 654:668 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 4 29 theme repeating 848:856 arg1 unit 858:861 the repeating unit 844:861 the repeating unit 844:861 The strategy was designed to suit the synthesis of oligomers of the repeating unit. 25674920 2 30 attach linked 505:510 arg2 functionality 491:503 a reactive functionality 480:503 a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 480:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 30 attach linked 505:510 arg1 end 539:541 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 515:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 31 with linkage 334:340 arg1 branch 297:302 a branch 295:302 a branch 295:302 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 0 32 theme repeating 26:34 arg1 unit 36:39 the repeating unit 22:39 the repeating unit of type Ia group B Streptococcus capsular polysaccharide 22:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 1 33 dep difficult 310:318 arg1 α-sialic 320:327 α-sialic 320:327 α-sialic 320:327 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 2 34 theme repeating 454:462 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 35 theme GBS 188:190 arg1 CPS 145:147 CPS 145:147 CPS 145:147 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 35 theme GBS 188:190 arg1 polysaccharide 129:142 the capsular polysaccharide 116:142 the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) 116:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 0 36 theme type 44:47 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 2 37 theme CPS 450:452 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 2 38 theme reactive 482:489 arg1 functionality 491:503 a reactive functionality 480:503 a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins 480:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 39 theme challenging 262:272 arg1 unit 245:248 its repeating unit 231:248 its repeating unit 231:248 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 39 theme challenging 262:272 arg1 pentasaccharide 274:288 a challenging pentasaccharide 260:288 a challenging pentasaccharide with a branch 260:302 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 2 40 with coupling 582:589 arg1 proteins 604:611 carrier proteins 596:611 carrier proteins 596:611 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 25674920 1 41 theme capsular 120:127 arg1 CPS 145:147 CPS 145:147 CPS 145:147 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 1 41 theme capsular 120:127 arg1 polysaccharide 129:142 the capsular polysaccharide 116:142 the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) 116:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 3 42 theme trisaccharide 753:765 arg1 acceptor 770:777 a branched trisaccharide as acceptor 742:777 a branched trisaccharide as acceptor 742:777 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 3 42 theme trisaccharide 753:765 arg1 sialo-disaccharide 710:727 a sialo-disaccharide 708:727 a sialo-disaccharide 708:727 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 0 43 theme group 52:56 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 1 44 theme polysaccharide 129:142 arg1 structure 103:111 The structure 99:111 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) 99:191 The structure of the capsular polysaccharide (CPS) of serotype Ia group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult α-sialic acid linkage, has not been synthesized yet. 25674920 3 45 theme as 767:768 arg1 acceptor 770:777 a branched trisaccharide as acceptor 742:777 a branched trisaccharide as acceptor 742:777 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 3 45 theme as 767:768 arg1 sialo-disaccharide 710:727 a sialo-disaccharide 708:727 a sialo-disaccharide 708:727 The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialo-disaccharide as donor and a branched trisaccharide as acceptor. 25674920 0 46 theme Ia 49:50 arg1 polysaccharide 83:96 type Ia group B Streptococcus capsular polysaccharide 44:96 type Ia group B Streptococcus capsular polysaccharide 44:96 Chemical synthesis of the repeating unit of type Ia group B Streptococcus capsular polysaccharide. 25674920 2 47 theme GBS 446:448 arg1 unit 464:467 the serotype Ia GBS CPS repeating unit 430:467 the serotype Ia GBS CPS repeating unit 430:467 In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. 28938401 9 0 theme metastasized 1688:1699 arg1 paragangliomas 1701:1714 metastasized paragangliomas 1688:1714 metastasized paragangliomas 1688:1714 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 6 1 theme tumor 1078:1082 arg1 tissue 1084:1089 tumor tissue 1078:1089 tumor tissue 1078:1089 Main Outcome Measure N-glycan profile of tumor tissue. 28938401 1 2 theme Context 109:115 arg1 methods 130:136 Context No effective methods 109:136 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential 109:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 7 3 from abundant 1150:1157 arg1 tumors 1175:1180 metastasized tumors 1162:1180 metastasized tumors 1162:1180 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 3 from abundant 1150:1157 arg1 tumors 1206:1211 nonmetastasized tumors 1190:1211 nonmetastasized tumors 1190:1211 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 4 gly multifucosylated 1295:1310 arg1 glycans 1312:1318 multifucosylated glycans 1295:1318 multifucosylated glycans (complex fucosylation) 1295:1341 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 4 gly multifucosylated 1295:1310 arg1 fucosylation 1329:1340 complex fucosylation 1321:1340 complex fucosylation 1321:1340 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 5 theme multifucosylated 1295:1310 arg1 glycans 1312:1318 multifucosylated glycans 1295:1318 multifucosylated glycans (complex fucosylation) 1295:1341 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 5 theme multifucosylated 1295:1310 arg1 fucosylation 1329:1340 complex fucosylation 1321:1340 complex fucosylation 1321:1340 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 9 6 theme nonmetastasized 1746:1760 arg1 tumors 1762:1767 nonmetastasized tumors 1746:1767 nonmetastasized tumors 1746:1767 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 8 7 theme acidic 1429:1434 arg1 N-glycans 1436:1444 acidic N-glycans 1429:1444 acidic N-glycans 1429:1444 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 10 8 theme MALDI-TOF 1892:1900 arg1 profiling 1905:1913 MALDI-TOF MS profiling 1892:1913 MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas 1892:1961 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 7 9 theme N-acetylglucosamine 1274:1292 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 2 10 theme metastasized 354:365 arg1 pheochromocytomas 367:383 metastasized pheochromocytomas 354:383 metastasized pheochromocytomas 354:383 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 4 11 theme metastasized 754:765 arg1 pheochromocytomas 767:783 8 primary metastasized pheochromocytomas 744:783 8 primary metastasized pheochromocytomas 744:783 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 7 12 theme cancer-associated 1247:1263 arg1 N-acetylglucosamine 1274:1292 cancer-associated terminal N-acetylglucosamine 1247:1292 cancer-associated terminal N-acetylglucosamine 1247:1292 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 0 13 theme Pheochromocytomas 24:40 arg1 Profiling 11:19 N-Glycomic Profiling 0:19 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas 0:59 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas Separates Metastatic and Nonmetastatic Disease. 28938401 7 14 theme nonmetastasized 1190:1204 arg1 tumors 1206:1211 nonmetastasized tumors 1190:1211 nonmetastasized tumors 1190:1211 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 5 15 theme tissue 1021:1026 arg1 samples 1028:1034 formalin-fixed, paraffin-embedded tissue samples 987:1034 formalin-fixed, paraffin-embedded tissue samples 987:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 10 16 theme primary 1918:1924 arg1 pheochromocytomas 1926:1942 primary pheochromocytomas 1918:1942 primary pheochromocytomas 1918:1942 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 8 17 gly multifucosylated 1489:1504 arg1 glycans 1506:1512 multifucosylated glycans 1489:1512 multifucosylated glycans 1489:1512 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 2 18 theme specific 271:278 arg1 structures 316:325 specific asparagine-linked glycan (N-glycan) structures 271:325 specific asparagine-linked glycan (N-glycan) structures 271:325 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 10 19 with diseases 1976:1983 arg1 potential 2001:2009 metastatic potential 1990:2009 metastatic potential 1990:2009 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 2 20 link asparagine-linked 280:296 arg1 structures 316:325 specific asparagine-linked glycan (N-glycan) structures 271:325 specific asparagine-linked glycan (N-glycan) structures 271:325 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 7 21 theme complex-type 1214:1225 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 4 22 dep pheochromocytomas 706:722 arg1 paragangliomas 788:801 paragangliomas 788:801 paragangliomas 788:801 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 4 22 dep pheochromocytomas 706:722 arg1 pheochromocytomas 767:783 8 primary metastasized pheochromocytomas 744:783 8 primary metastasized pheochromocytomas 744:783 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 2 23 theme structures 316:325 arg1 tool 431:434 a diagnostic tool 418:434 a diagnostic tool 418:434 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 2 23 theme structures 316:325 arg1 identification 253:266 The identification 249:266 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, 249:403 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 7 24 theme metastasized 1162:1173 arg1 tumors 1175:1180 metastasized tumors 1162:1180 metastasized tumors 1162:1180 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 4 25 theme nonmetastasized 809:823 arg1 tumors 825:830 8 nonmetastasized tumors 807:830 8 nonmetastasized tumors 807:830 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 11 26 theme N-glycan 2079:2086 arg1 structures 2088:2097 malignancy-linked N-glycan structures 2061:2097 malignancy-linked N-glycan structures 2061:2097 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 11 26 theme N-glycan 2079:2086 arg1 tools 2133:2137 potential diagnostic tools 2112:2137 potential diagnostic tools for pheochromocytomas and paragangliomas 2112:2178 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 2 27 theme N-glycan 306:313 arg1 structures 316:325 specific asparagine-linked glycan (N-glycan) structures 271:325 specific asparagine-linked glycan (N-glycan) structures 271:325 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 9 28 theme positivity 1814:1823 arg1 rate 1834:1837 the false positivity critical rate 1804:1837 the false positivity critical rate 1804:1837 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 11 29 theme diagnostic 2122:2131 arg1 structures 2088:2097 malignancy-linked N-glycan structures 2061:2097 malignancy-linked N-glycan structures 2061:2097 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 11 29 theme diagnostic 2122:2131 arg1 tools 2133:2137 potential diagnostic tools 2112:2137 potential diagnostic tools for pheochromocytomas and paragangliomas 2112:2178 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 5 30 theme MALDI-TOF 940:948 arg1 spectrometry 956:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry 880:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples 880:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 30 theme MALDI-TOF 940:948 arg1 MS 970:971 MS 970:971 MS 970:971 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 1 31 theme metastatic 203:212 arg1 potential 214:222 metastatic potential 203:222 metastatic potential 203:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 8 32 theme hybrid-type/monoantennary 1577:1601 arg1 N-glycans 1603:1611 hybrid-type/monoantennary N-glycans 1577:1611 hybrid-type/monoantennary N-glycans 1577:1611 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 1 33 with paragangliomas 183:196 arg1 potential 214:222 metastatic potential 203:222 metastatic potential 203:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 7 34 theme N-glycan 1123:1130 arg1 signals 1132:1138 neutral N-glycan signals 1115:1138 neutral N-glycan signals 1115:1138 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 5 35 theme laser 896:900 arg1 spectrometry 956:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry 880:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples 880:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 35 theme laser 896:900 arg1 MS 970:971 MS 970:971 MS 970:971 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 1 36 with pheochromocytomas 161:177 arg1 potential 214:222 metastatic potential 203:222 metastatic potential 203:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 0 37 theme Nonmetastatic 86:98 arg1 Disease 100:106 Metastatic and Nonmetastatic Disease 71:106 Metastatic and Nonmetastatic Disease 71:106 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas Separates Metastatic and Nonmetastatic Disease. 28938401 3 38 theme primary 497:503 arg1 metastasized 505:516 primary metastasized and nonmetastasized pheochromocytomas 497:554 metastasized 505:516 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 0 39 theme N-Glycomic 0:9 arg1 Profiling 11:19 N-Glycomic Profiling 0:19 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas 0:59 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas Separates Metastatic and Nonmetastatic Disease. 28938401 1 40 theme No 117:118 arg1 methods 130:136 Context No effective methods 109:136 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential 109:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 8 41 dep abundant 1456:1463 arg1 N-glycans 1603:1611 hybrid-type/monoantennary N-glycans 1577:1611 hybrid-type/monoantennary N-glycans 1577:1611 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 8 41 dep abundant 1456:1463 arg1 glycans 1506:1512 multifucosylated glycans 1489:1512 multifucosylated glycans 1489:1512 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 8 41 dep abundant 1456:1463 arg1 glycans 1564:1570 acid ester-modified (sulfated or phosphorylated) glycans 1515:1570 acid ester-modified (sulfated or phosphorylated) glycans 1515:1570 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 7 42 theme pauci-mannose-type 1383:1400 arg1 N-glycans 1402:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 6 43 theme Outcome 1042:1048 arg1 Measure 1050:1056 Main Outcome Measure 1037:1056 Main Outcome Measure 1037:1056 Main Outcome Measure N-glycan profile of tumor tissue. 28938401 3 44 theme pheochromocytomas 538:554 arg1 paragangliomas 560:573 paragangliomas 560:573 paragangliomas 560:573 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 3 44 theme pheochromocytomas 538:554 arg1 profiles 485:492 N-glycomic profiles 474:492 N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas 474:554 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 5 45 theme mass 951:954 arg1 spectrometry 956:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry 880:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples 880:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 45 theme mass 951:954 arg1 MS 970:971 MS 970:971 MS 970:971 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 7 46 theme N-glycans 1356:1364 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 9 47 theme complex 1631:1637 arg1 fucosylation 1639:1650 complex fucosylation 1631:1650 complex fucosylation 1631:1650 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 9 48 from abundant 1676:1683 arg1 pheochromocytomas 1720:1736 pheochromocytomas 1720:1736 pheochromocytomas 1720:1736 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 9 48 from abundant 1676:1683 arg1 paragangliomas 1701:1714 metastasized paragangliomas 1688:1714 metastasized paragangliomas 1688:1714 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 9 48 from abundant 1676:1683 arg1 tumors 1762:1767 nonmetastasized tumors 1746:1767 nonmetastasized tumors 1746:1767 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 9 49 from tumors 1762:1767 arg1 abundant 1676:1683 abundant 1676:1683 abundant 1676:1683 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 11 50 theme malignancy-linked 2061:2077 arg1 structures 2088:2097 malignancy-linked N-glycan structures 2061:2097 malignancy-linked N-glycan structures 2061:2097 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 11 50 theme malignancy-linked 2061:2077 arg1 tools 2133:2137 potential diagnostic tools 2112:2137 potential diagnostic tools for pheochromocytomas and paragangliomas 2112:2178 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 10 51 theme different 2026:2034 arg1 profiles 2045:2052 their different N-glycan profiles 2020:2052 their different N-glycan profiles 2020:2052 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 5 52 dep formalin-fixed 987:1000 arg1 paraffin-embedded 1003:1019 paraffin-embedded 1003:1019 paraffin-embedded 1003:1019 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 7 53 theme hybrid-type 1344:1354 arg1 N-glycans 1356:1364 hybrid-type N-glycans 1344:1364 hybrid-type N-glycans 1344:1364 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 8 54 dep ester-modified 1520:1533 arg1 sulfated 1536:1543 sulfated 1536:1543 sulfated 1536:1543 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 8 54 dep ester-modified 1520:1533 arg1 phosphorylated 1548:1561 phosphorylated 1548:1561 phosphorylated 1548:1561 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 9 55 from pheochromocytomas 1720:1736 arg1 abundant 1676:1683 abundant 1676:1683 abundant 1676:1683 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 2 56 theme diagnostic 420:429 arg1 identification 253:266 The identification 249:266 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, 249:403 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 2 56 theme diagnostic 420:429 arg1 tool 431:434 a diagnostic tool 418:434 a diagnostic tool 418:434 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 6 57 theme tissue 1084:1089 arg1 profile 1067:1073 profile 1067:1073 profile of tumor tissue 1067:1089 Main Outcome Measure N-glycan profile of tumor tissue. 28938401 7 58 theme complex 1321:1327 arg1 glycans 1312:1318 multifucosylated glycans 1295:1318 multifucosylated glycans (complex fucosylation) 1295:1341 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 58 theme complex 1321:1327 arg1 fucosylation 1329:1340 complex fucosylation 1321:1340 complex fucosylation 1321:1340 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 8 59 from abundant 1456:1463 arg1 tumors 1481:1486 metastasized tumors 1468:1486 metastasized tumors 1468:1486 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 7 60 theme glycans 1312:1318 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 61 dep Results 1092:1098 arg1 abundant 1150:1157 abundant 1150:1157 abundant 1150:1157 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 62 from tumors 1175:1180 arg1 abundant 1150:1157 abundant 1150:1157 abundant 1150:1157 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 10 63 theme MS 1902:1903 arg1 profiling 1905:1913 MALDI-TOF MS profiling 1892:1913 MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas 1892:1961 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 5 64 theme Methods 833:839 arg1 structures 850:859 Methods N-glycan structures 833:859 Methods N-glycan structures 833:859 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 65 theme formalin-fixed 987:1000 arg1 samples 1028:1034 formalin-fixed, paraffin-embedded tissue samples 987:1034 formalin-fixed, paraffin-embedded tissue samples 987:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 8 66 theme multifucosylated 1489:1504 arg1 glycans 1506:1512 multifucosylated glycans 1489:1512 multifucosylated glycans 1489:1512 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 10 67 theme paragangliomas 1948:1961 arg1 profiling 1905:1913 MALDI-TOF MS profiling 1892:1913 MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas 1892:1961 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 10 68 dep Conclusions 1880:1890 arg1 identify 1967:1974 identify 1967:1974 can identify diseases with metastatic potential based on their different N-glycan profiles. 1963:2053 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 9 69 theme individual 1773:1782 arg1 tests 1784:1788 individual tests 1773:1788 individual tests 1773:1788 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 8 70 theme N-glycans 1436:1444 arg1 groups 1419:1424 Three groups 1413:1424 Three groups of acidic N-glycans 1413:1444 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 8 70 theme N-glycans 1436:1444 arg1 N-glycans 1436:1444 acidic N-glycans 1429:1444 acidic N-glycans 1429:1444 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 4 71 theme primary 746:752 arg1 pheochromocytomas 767:783 8 primary metastasized pheochromocytomas 744:783 8 primary metastasized pheochromocytomas 744:783 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 10 72 theme pheochromocytomas 1926:1942 arg1 profiling 1905:1913 MALDI-TOF MS profiling 1892:1913 MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas 1892:1961 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 7 73 theme N-glycan 1227:1234 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 2 74 theme asparagine-linked 280:296 arg1 structures 316:325 specific asparagine-linked glycan (N-glycan) structures 271:325 specific asparagine-linked glycan (N-glycan) structures 271:325 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 8 75 from tumors 1481:1486 arg1 abundant 1456:1463 abundant 1456:1463 abundant 1456:1463 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 7 76 theme terminal 1265:1272 arg1 N-acetylglucosamine 1274:1292 cancer-associated terminal N-acetylglucosamine 1247:1292 cancer-associated terminal N-acetylglucosamine 1247:1292 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 8 77 theme ester-modified 1520:1533 arg1 glycans 1564:1570 acid ester-modified (sulfated or phosphorylated) glycans 1515:1570 acid ester-modified (sulfated or phosphorylated) glycans 1515:1570 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 1 78 theme primary 153:159 arg1 pheochromocytomas 161:177 primary pheochromocytomas 153:177 primary pheochromocytomas 153:177 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 5 79 theme desorption-ionization 902:922 arg1 spectrometry 956:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry 880:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples 880:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 79 theme desorption-ionization 902:922 arg1 MS 970:971 MS 970:971 MS 970:971 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 9 80 theme false 1808:1812 arg1 rate 1834:1837 the false positivity critical rate 1804:1837 the false positivity critical rate 1804:1837 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 9 81 theme critical 1825:1832 arg1 rate 1834:1837 the false positivity critical rate 1804:1837 the false positivity critical rate 1804:1837 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 5 82 theme N-glycan 841:848 arg1 structures 850:859 Methods N-glycan structures 833:859 Methods N-glycan structures 833:859 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 11 83 theme potential 2112:2120 arg1 structures 2088:2097 malignancy-linked N-glycan structures 2061:2097 malignancy-linked N-glycan structures 2061:2097 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 11 83 theme potential 2112:2120 arg1 tools 2133:2137 potential diagnostic tools 2112:2137 potential diagnostic tools for pheochromocytomas and paragangliomas 2112:2178 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 5 84 theme matrix-assisted 880:894 arg1 spectrometry 956:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry 880:967 matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples 880:1034 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 5 84 theme matrix-assisted 880:894 arg1 MS 970:971 MS 970:971 MS 970:971 Methods N-glycan structures were analyzed with matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) profiling of formalin-fixed, paraffin-embedded tissue samples. 28938401 7 85 theme signals 1132:1138 arg1 signals 1132:1138 neutral N-glycan signals 1115:1138 neutral N-glycan signals 1115:1138 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 7 85 theme signals 1132:1138 arg1 groups 1105:1110 Four groups 1100:1110 Four groups of neutral N-glycan signals 1100:1138 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 3 86 theme nonmetastasized 522:536 arg1 pheochromocytomas 538:554 primary metastasized and nonmetastasized pheochromocytomas 497:554 pheochromocytomas 538:554 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 7 87 theme N-glycans 1402:1410 arg1 signals 1236:1242 complex-type N-glycan signals 1214:1242 complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans 1214:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 3 88 from differences 459:469 arg1 paragangliomas 560:573 paragangliomas 560:573 paragangliomas 560:573 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 3 88 from differences 459:469 arg1 profiles 485:492 N-glycomic profiles 474:492 N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas 474:554 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 7 89 theme neutral 1115:1121 arg1 signals 1132:1138 neutral N-glycan signals 1115:1138 neutral N-glycan signals 1115:1138 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 9 90 theme multiplicity 1858:1869 arg1 testing 1871:1877 multiplicity testing 1858:1877 multiplicity testing 1858:1877 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 8 91 theme metastasized 1468:1479 arg1 tumors 1481:1486 metastasized tumors 1468:1486 metastasized tumors 1468:1486 Three groups of acidic N-glycans were more abundant in metastasized tumors: multifucosylated glycans, acid ester-modified (sulfated or phosphorylated) glycans, and hybrid-type/monoantennary N-glycans. 28938401 3 92 theme metastasized 505:516 arg1 paragangliomas 560:573 paragangliomas 560:573 paragangliomas 560:573 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 3 92 theme metastasized 505:516 arg1 profiles 485:492 N-glycomic profiles 474:492 N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas 474:554 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 7 93 theme fucosylated 1371:1381 arg1 N-glycans 1402:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 9 94 from paragangliomas 1701:1714 arg1 abundant 1676:1683 abundant 1676:1683 abundant 1676:1683 Fucosylation and complex fucosylation were significantly more abundant in metastasized paragangliomas and pheochromocytomas than in nonmetastasized tumors for individual tests but were over the false positivity critical rate, when adjusted for multiplicity testing. 28938401 1 95 theme effective 120:128 arg1 methods 130:136 Context No effective methods 109:136 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential 109:222 Context No effective methods for separating primary pheochromocytomas and paragangliomas with metastatic potential are currently available. 28938401 0 96 theme Paragangliomas 46:59 arg1 Profiling 11:19 N-Glycomic Profiling 0:19 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas 0:59 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas Separates Metastatic and Nonmetastatic Disease. 28938401 4 97 dep Setting 576:582 arg1 included 694:701 included 694:701 included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors 694:830 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 4 97 dep Setting 576:582 arg1 conducted 599:607 conducted 599:607 was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. 595:688 Setting This study was conducted at Helsinki University Hospital, University of Helsinki, and Glykos Finland Ltd. and included 16 pheochromocytomas and paragangliomas: 8 primary metastasized pheochromocytomas or paragangliomas and 8 nonmetastasized tumors. 28938401 6 98 theme Main 1037:1040 arg1 Measure 1050:1056 Main Outcome Measure 1037:1056 Main Outcome Measure 1037:1056 Main Outcome Measure N-glycan profile of tumor tissue. 28938401 10 99 theme metastatic 1990:1999 arg1 potential 2001:2009 metastatic potential 1990:2009 metastatic potential 1990:2009 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 0 100 theme Metastatic 71:80 arg1 Disease 100:106 Metastatic and Nonmetastatic Disease 71:106 Metastatic and Nonmetastatic Disease 71:106 N-Glycomic Profiling of Pheochromocytomas and Paragangliomas Separates Metastatic and Nonmetastatic Disease. 28938401 3 101 theme N-glycomic 474:483 arg1 profiles 485:492 N-glycomic profiles 474:492 N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas 474:554 Objective To identify differences in N-glycomic profiles of primary metastasized and nonmetastasized pheochromocytomas and paragangliomas. 28938401 10 102 theme N-glycan 2036:2043 arg1 profiles 2045:2052 their different N-glycan profiles 2020:2052 their different N-glycan profiles 2020:2052 Conclusions MALDI-TOF MS profiling of primary pheochromocytomas and paragangliomas can identify diseases with metastatic potential based on their different N-glycan profiles. 28938401 2 103 theme glycan 298:303 arg1 structures 316:325 specific asparagine-linked glycan (N-glycan) structures 271:325 specific asparagine-linked glycan (N-glycan) structures 271:325 The identification of specific asparagine-linked glycan (N-glycan) structures, which are associated with metastasized pheochromocytomas and paragangliomas, may serve as a diagnostic tool. 28938401 7 104 from tumors 1206:1211 arg1 abundant 1150:1157 abundant 1150:1157 abundant 1150:1157 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 28938401 11 105 link malignancy-linked 2061:2077 arg1 structures 2088:2097 malignancy-linked N-glycan structures 2061:2097 malignancy-linked N-glycan structures 2061:2097 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 11 105 link malignancy-linked 2061:2077 arg1 tools 2133:2137 potential diagnostic tools 2112:2137 potential diagnostic tools for pheochromocytomas and paragangliomas 2112:2178 Thus, malignancy-linked N-glycan structures may serve as potential diagnostic tools for pheochromocytomas and paragangliomas. 28938401 7 106 gly fucosylated 1371:1381 arg1 N-glycans 1402:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 fucosylated pauci-mannose-type N-glycans 1371:1410 Results Four groups of neutral N-glycan signals were more abundant in metastasized tumors than in nonmetastasized tumors: complex-type N-glycan signals of cancer-associated terminal N-acetylglucosamine, multifucosylated glycans (complex fucosylation), hybrid-type N-glycans, and fucosylated pauci-mannose-type N-glycans. 27259834 6 0 theme β-1,3-N-acetylglucosaminyltransferase 966:1002 arg1 gene 1016:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 10 1 theme 2+ 1524:1525 arg1 concentration 1528:1540 CONCLUSIONS Physiological Mn(2+) concentration 1495:1540 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae 1495:1557 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 11 2 theme Mn 1619:1620 arg1 supplementation 1626:1640 Mn(2+) supplementation 1619:1640 Mn(2+) supplementation 1619:1640 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 2 3 theme mucin-type 382:391 arg1 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 2 3 theme mucin-type 382:391 arg1 structure 400:408 mucin-type core 1 structure 382:408 mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 382:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 1 4 theme useful 293:298 arg1 glycoproteins 300:312 clinically useful glycoproteins 282:312 clinically useful glycoproteins 282:312 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 6 5 theme codon-optimized 934:948 arg1 UDP-GlcNAc 950:959 a codon-optimized UDP-GlcNAc 932:959 a codon-optimized UDP-GlcNAc 932:959 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 3 6 theme core 574:577 arg1 strain 591:596 core 3-producing strain 574:596 core 3-producing strain 574:596 METHODS To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. 27259834 10 7 theme optimal 1590:1596 arg1 synthesis 1598:1606 optimal synthesis 1590:1606 optimal synthesis of core 3 1590:1616 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 1 8 gly glycoproteins 233:245 arg1 glycoproteins 233:245 disease-related glycoproteins 217:245 disease-related glycoproteins 217:245 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 2 9 theme capable 349:355 arg1 strain 342:347 A Saccharomyces cerevisiae strain 315:347 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 315:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 1 10 theme specific 205:212 arg1 antibodies 194:203 antibodies 194:203 antibodies specific to disease-related glycoproteins 194:245 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 8 11 theme % 1283:1283 arg1 increases 1262:1270 increases 1262:1270 increases of up to 25% in the accumulation of core 3 on the MUC1ap 1262:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 1 12 theme disease-related 217:231 arg1 glycoproteins 233:245 disease-related glycoproteins 217:245 disease-related glycoproteins 217:245 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 8 13 theme culture 1239:1245 arg1 medium 1247:1252 the culture medium 1235:1252 the culture medium 1235:1252 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 7 14 theme highest 1113:1119 arg1 activity 1130:1137 The highest in vitro activity 1109:1137 The highest in vitro activity of β3Gn-T6 1109:1148 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 7 15 theme 10mM 1191:1194 arg1 concentrations 1173:1186 Mn(2+) concentrations 1166:1186 Mn(2+) concentrations of 10mM and above 1166:1204 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 9 16 theme MnCl2 1474:1478 arg1 addition 1462:1469 the addition 1458:1469 the addition of MnCl2 to the medium 1458:1492 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 1 17 theme various 126:132 arg1 glycoproteins 145:157 various mucin-like glycoproteins 126:157 various mucin-like glycoproteins 126:157 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 4 18 theme acceptor 785:792 arg1 MUC1ap 803:808 MUC1ap 803:808 MUC1ap 803:808 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 18 theme acceptor 785:792 arg1 peptide 794:800 A Mucin-1-derived acceptor peptide 767:800 A Mucin-1-derived acceptor peptide (MUC1ap) 767:809 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 18 theme acceptor 785:792 arg1 acceptor 831:838 an acceptor 828:838 an acceptor 828:838 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 3 19 theme 3-producing 579:589 arg1 strain 591:596 core 3-producing strain 574:596 core 3-producing strain 574:596 METHODS To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. 27259834 1 20 theme glycoproteins 145:157 arg1 Production 112:121 BACKGROUND Production 101:121 BACKGROUND Production of various mucin-like glycoproteins 101:157 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 2 21 dep Saccharomyces 317:329 arg1 cerevisiae 331:340 cerevisiae 331:340 cerevisiae 331:340 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 11 22 theme production 1739:1748 arg1 increases 1719:1727 increases 1719:1727 increases of core 3 production 1719:1748 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 9 23 from strain 1376:1381 arg1 invertase 1340:1348 The yeast invertase 1330:1348 The yeast invertase from the core 3-producing strain 1330:1381 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 9 23 from strain 1376:1381 arg1 N-glycosylated 1404:1417 N-glycosylated 1404:1417 N-glycosylated 1404:1417 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 7 24 located observed 1154:1161 arg1 concentrations 1173:1186 Mn(2+) concentrations 1166:1186 Mn(2+) concentrations of 10mM and above 1166:1204 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 7 24 located observed 1154:1161 arg2 activity 1130:1137 The highest in vitro activity 1109:1137 The highest in vitro activity of β3Gn-T6 1109:1148 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 11 25 theme core 1732:1735 arg1 production 1739:1748 core 3 production 1732:1748 core 3 production 1732:1748 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 8 26 theme core 1308:1311 arg1 accumulation 1292:1303 the accumulation 1288:1303 the accumulation of core 3 on the MUC1ap 1288:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 8 27 dep 25 1281:1282 arg1 to 1278:1279 to 1278:1279 to 1278:1279 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 2 28 theme core 480:483 arg1 structure 487:495 core 3 structure 480:495 core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) 480:526 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 2 28 theme core 480:483 arg1 GlcNAcβ1-3GalNAcα1-O-Ser/Thr 498:525 GlcNAcβ1-3GalNAcα1-O-Ser/Thr 498:525 GlcNAcβ1-3GalNAcα1-O-Ser/Thr 498:525 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 0 29 from production 36:45 arg1 cerevisiae 89:98 Saccharomyces cerevisiae 75:98 Saccharomyces cerevisiae 75:98 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 0 30 theme Manganese 0:8 arg1 concentration 14:26 Manganese ion concentration 0:26 Manganese ion concentration 0:26 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 1 31 theme glycoproteins 300:312 arg1 biosynthesis 266:277 the biosynthesis 262:277 the biosynthesis of clinically useful glycoproteins 262:312 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 12 32 theme specific 1859:1866 arg1 glycans 1883:1889 specific mammalian-type glycans 1859:1889 specific mammalian-type glycans 1859:1889 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 6 33 from increases 1029:1037 arg1 activity 1050:1057 β3Gn-T6 activity 1042:1057 β3Gn-T6 activity 1042:1057 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 5 34 theme modified 869:876 arg1 peptide 878:884 the resulting modified peptide 855:884 the resulting modified peptide 855:884 The amount of the resulting modified peptide was analyzed by HPLC. 27259834 12 35 theme concentration 1814:1826 arg1 control 1796:1802 control 1796:1802 control of Mn(2+) concentration 1796:1826 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 6 36 theme production 1097:1106 arg1 level 1081:1085 the level 1077:1085 the level of core 3 production 1077:1106 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 11 37 theme glycosylation 1678:1690 arg1 reaction 1666:1673 reaction 1666:1673 reaction of glycosylation 1666:1690 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 6 38 theme core 1090:1093 arg1 production 1097:1106 core 3 production 1090:1106 core 3 production 1090:1106 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 3 39 theme UDP-GlcNAc 681:690 arg1 transporter 692:702 UDP-GlcNAc transporter 681:702 UDP-GlcNAc transporter 681:702 METHODS To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. 27259834 9 40 gly N-glycosylated 1404:1417 arg1 invertase 1340:1348 The yeast invertase 1330:1348 The yeast invertase from the core 3-producing strain 1330:1381 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 9 40 gly N-glycosylated 1404:1417 arg1 N-glycosylated 1404:1417 N-glycosylated 1404:1417 N-glycosylated 1404:1417 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 0 41 theme human 50:54 arg1 O-glycan 63:70 human core 3 O-glycan 50:70 human core 3 O-glycan 50:70 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 9 42 theme 3-producing 1364:1374 arg1 strain 1376:1381 the core 3-producing strain 1355:1381 the core 3-producing strain 1355:1381 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 12 43 theme Mn 1807:1808 arg1 concentration 1814:1826 Mn(2+) concentration 1807:1826 Mn(2+) concentration 1807:1826 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 6 44 dep RESULTS 908:914 arg1 yielded 1021:1027 yielded 1021:1027 yielded increases in β3Gn-T6 activity 1021:1057 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 6 44 dep RESULTS 908:914 arg1 alter 1071:1075 alter 1071:1075 did not alter the level of core 3 production 1063:1106 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 6 44 dep RESULTS 908:914 arg1 Introduction 916:927 Introduction 916:927 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 908:1107 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 10 45 theme Physiological 1507:1519 arg1 concentration 1528:1540 CONCLUSIONS Physiological Mn(2+) concentration 1495:1540 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae 1495:1557 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 12 46 theme 2+ 1810:1811 arg1 concentration 1814:1826 Mn(2+) concentration 1807:1826 Mn(2+) concentration 1807:1826 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 2 47 theme structure 400:408 arg1 production 368:377 in vivo production 360:377 in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 360:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 7 48 dep in 1121:1122 arg1 vitro 1124:1128 vitro 1124:1128 vitro 1124:1128 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 7 49 theme Mn 1166:1167 arg1 concentrations 1173:1186 Mn(2+) concentrations 1166:1186 Mn(2+) concentrations of 10mM and above 1166:1204 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 2 50 theme core 393:396 arg1 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 Galβ1-3GalNAcα1-O-Ser/Thr 411:435 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 2 50 theme core 393:396 arg1 structure 400:408 mucin-type core 1 structure 382:408 mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 382:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 6 51 theme βGal 961:964 arg1 gene 1016:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 11 52 theme reaction 1666:1673 arg1 up-regulation 1649:1661 up-regulation 1649:1661 up-regulation of reaction of glycosylation in the Golgi 1649:1703 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 2 53 theme Saccharomyces 317:329 arg1 strain 342:347 A Saccharomyces cerevisiae strain 315:347 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 315:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 6 54 theme UDP-GlcNAc 950:959 arg1 Introduction 916:927 Introduction 916:927 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 908:1107 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 4 55 link Mucin-1-derived 769:783 arg1 MUC1ap 803:808 MUC1ap 803:808 MUC1ap 803:808 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 55 link Mucin-1-derived 769:783 arg1 peptide 794:800 A Mucin-1-derived acceptor peptide 767:800 A Mucin-1-derived acceptor peptide (MUC1ap) 767:809 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 55 link Mucin-1-derived 769:783 arg1 acceptor 831:838 an acceptor 828:838 an acceptor 828:838 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 7 56 theme β3Gn-T6 1142:1148 arg1 activity 1130:1137 The highest in vitro activity 1109:1137 The highest in vitro activity of β3Gn-T6 1109:1148 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 1 57 theme antibodies 194:203 arg1 development 179:189 development 179:189 development of antibodies specific to disease-related glycoproteins 179:245 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 6 58 theme β3Gn-T6 1042:1048 arg1 activity 1050:1057 β3Gn-T6 activity 1042:1057 β3Gn-T6 activity 1042:1057 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 12 59 theme glycans 1883:1889 arg1 production 1845:1854 production 1845:1854 production of specific mammalian-type glycans in S. cerevisiae 1845:1906 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 7 60 theme in 1121:1122 arg1 activity 1130:1137 The highest in vitro activity 1109:1137 The highest in vitro activity of β3Gn-T6 1109:1148 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 27259834 3 61 dep -Gal-4-epimerase 639:654 arg1 UDP-GalNAc 657:666 UDP-GalNAc 657:666 UDP-GalNAc 657:666 METHODS To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. 27259834 8 62 theme MnCl2 1226:1230 arg1 Supplementation 1207:1221 Supplementation 1207:1221 Supplementation of MnCl2 to the culture medium 1207:1252 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 1 63 gly glycoproteins 145:157 arg1 glycoproteins 145:157 various mucin-like glycoproteins 126:157 various mucin-like glycoproteins 126:157 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 10 64 theme core 1611:1614 arg1 synthesis 1598:1606 optimal synthesis 1590:1606 optimal synthesis of core 3 1590:1616 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 1 65 gly glycoproteins 300:312 arg1 glycoproteins 300:312 clinically useful glycoproteins 282:312 clinically useful glycoproteins 282:312 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 6 66 theme β3Gn-T6 1007:1013 arg1 gene 1016:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene 961:1019 RESULTS Introduction of a codon-optimized UDP-GlcNAc:βGal β-1,3-N-acetylglucosaminyltransferase 6 (β3Gn-T6) gene yielded increases in β3Gn-T6 activity but did not alter the level of core 3 production. 27259834 8 67 from increases 1262:1270 arg1 accumulation 1292:1303 the accumulation 1288:1303 the accumulation of core 3 on the MUC1ap 1288:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 8 67 from increases 1262:1270 arg1 MUC1ap 1322:1327 the MUC1ap 1318:1327 the MUC1ap 1318:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 1 68 theme mucin-like 134:143 arg1 glycoproteins 145:157 various mucin-like glycoproteins 126:157 various mucin-like glycoproteins 126:157 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 4 69 theme Mucin-1-derived 769:783 arg1 MUC1ap 803:808 MUC1ap 803:808 MUC1ap 803:808 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 69 theme Mucin-1-derived 769:783 arg1 peptide 794:800 A Mucin-1-derived acceptor peptide 767:800 A Mucin-1-derived acceptor peptide (MUC1ap) 767:809 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 4 69 theme Mucin-1-derived 769:783 arg1 acceptor 831:838 an acceptor 828:838 an acceptor 828:838 A Mucin-1-derived acceptor peptide (MUC1ap) was expressed as an acceptor. 27259834 0 70 theme O-glycan 63:70 arg1 production 36:45 production 36:45 production of human core 3 O-glycan in Saccharomyces cerevisiae 36:98 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 3 71 theme -Gal-4-epimerase 639:654 arg1 transporter 668:678 uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter 614:678 uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter 614:678 METHODS To generate core 3-producing strain, genes encoding uridine diphosphate (UDP)-Gal-4-epimerase, UDP-GalNAc transporter, UDP-GlcNAc transporter, and two glycosyltransferases were integrated into the genome. 27259834 5 72 theme peptide 878:884 arg1 peptide 878:884 the resulting modified peptide 855:884 the resulting modified peptide 855:884 The amount of the resulting modified peptide was analyzed by HPLC. 27259834 5 72 theme peptide 878:884 arg1 amount 845:850 The amount 841:850 The amount of the resulting modified peptide 841:884 The amount of the resulting modified peptide was analyzed by HPLC. 27259834 0 73 theme Saccharomyces 75:87 arg1 cerevisiae 89:98 Saccharomyces cerevisiae 75:98 Saccharomyces cerevisiae 75:98 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 8 74 from accumulation 1292:1303 arg1 MUC1ap 1322:1327 the MUC1ap 1318:1327 the MUC1ap 1318:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 12 75 from production 1845:1854 arg1 cerevisiae 1897:1906 S. cerevisiae 1894:1906 S. cerevisiae 1894:1906 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 12 76 dep SIGNIFICANCE 1759:1770 arg1 reveals 1783:1789 reveals 1783:1789 reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae 1783:1906 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 0 77 theme ion 10:12 arg1 concentration 14:26 Manganese ion concentration 0:26 Manganese ion concentration 0:26 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 12 78 theme mammalian-type 1868:1881 arg1 glycans 1883:1889 specific mammalian-type glycans 1859:1889 specific mammalian-type glycans 1859:1889 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 8 79 from MUC1ap 1322:1327 arg1 increases 1262:1270 increases 1262:1270 increases of up to 25% in the accumulation of core 3 on the MUC1ap 1262:1327 Supplementation of MnCl2 to the culture medium yielded increases of up to 25% in the accumulation of core 3 on the MUC1ap. 27259834 2 80 dep in 360:361 arg1 vivo 363:366 vivo 363:366 vivo 363:366 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 5 81 theme resulting 859:867 arg1 peptide 878:884 the resulting modified peptide 855:884 the resulting modified peptide 855:884 The amount of the resulting modified peptide was analyzed by HPLC. 27259834 9 82 theme yeast 1334:1338 arg1 invertase 1340:1348 The yeast invertase 1330:1348 The yeast invertase from the core 3-producing strain 1330:1381 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 9 82 theme yeast 1334:1338 arg1 N-glycosylated 1404:1417 N-glycosylated 1404:1417 N-glycosylated 1404:1417 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 1 83 theme BACKGROUND 101:110 arg1 Production 112:121 BACKGROUND Production 101:121 BACKGROUND Production of various mucin-like glycoproteins 101:157 BACKGROUND Production of various mucin-like glycoproteins could be useful for development of antibodies specific to disease-related glycoproteins as well as for the biosynthesis of clinically useful glycoproteins. 27259834 11 84 from up-regulation 1649:1661 arg1 Golgi 1699:1703 the Golgi 1695:1703 the Golgi 1695:1703 Mn(2+) supplementation led to up-regulation of reaction of glycosylation in the Golgi, resulting in increases of core 3 production. 27259834 0 85 theme core 56:59 arg1 O-glycan 63:70 human core 3 O-glycan 50:70 human core 3 O-glycan 50:70 Manganese ion concentration affects production of human core 3 O-glycan in Saccharomyces cerevisiae. 27259834 9 86 theme core 1359:1362 arg1 strain 1376:1381 the core 3-producing strain 1355:1381 the core 3-producing strain 1355:1381 The yeast invertase from the core 3-producing strain was less extensively N-glycosylated; however, it was partially restored by the addition of MnCl2 to the medium. 27259834 10 87 from concentration 1528:1540 arg1 cerevisiae 1548:1557 S. cerevisiae 1545:1557 S. cerevisiae 1545:1557 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 10 88 theme Mn 1521:1522 arg1 concentration 1528:1540 CONCLUSIONS Physiological Mn(2+) concentration 1495:1540 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae 1495:1557 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 2 89 theme production 368:377 arg1 capable 349:355 capable 349:355 capable 349:355 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 10 90 theme CONCLUSIONS 1495:1505 arg1 concentration 1528:1540 CONCLUSIONS Physiological Mn(2+) concentration 1495:1540 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae 1495:1557 CONCLUSIONS Physiological Mn(2+) concentration in S. cerevisiae was insufficient to facilitate optimal synthesis of core 3. 27259834 2 91 theme in 360:361 arg1 production 368:377 in vivo production 360:377 in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) 360:436 A Saccharomyces cerevisiae strain capable of in vivo production of mucin-type core 1 structure (Galβ1-3GalNAcα1-O-Ser/Thr) has been reported, but a strain producing core 3 structure (GlcNAcβ1-3GalNAcα1-O-Ser/Thr) has not been constructed. 27259834 12 92 theme GENERAL 1751:1757 arg1 SIGNIFICANCE 1759:1770 GENERAL SIGNIFICANCE 1751:1770 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 1751:1907 GENERAL SIGNIFICANCE This study reveals that control of Mn(2+) concentration is important for production of specific mammalian-type glycans in S. cerevisiae. 27259834 7 93 theme above 1200:1204 arg1 concentrations 1173:1186 Mn(2+) concentrations 1166:1186 Mn(2+) concentrations of 10mM and above 1166:1204 The highest in vitro activity of β3Gn-T6 was observed at Mn(2+) concentrations of 10mM and above. 26595660 2 0 theme system 490:495 arg1 protection 467:476 a temporary protection 455:476 a temporary protection of the diol system 455:495 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 0 theme system 490:495 arg1 glycosylation 295:307 highly regioselective glycosylation 273:307 highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 273:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 1 theme diol 485:488 arg1 system 490:495 the diol system 481:495 the diol system 481:495 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 3 2 theme deprotection 515:526 arg1 steps 546:550 the deprotection and glycosylation steps 511:550 steps 546:550 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 4 3 theme ones 734:737 arg1 presence 714:721 the presence 710:721 the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate 710:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 4 4 theme benzoyl 726:732 arg1 ones 734:737 benzoyl ones 726:737 benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate 726:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 2 5 theme synthetic 247:255 arg1 scheme 257:262 The synthetic scheme 243:262 The synthetic scheme 243:262 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 4 6 theme selective 672:680 arg1 removal 682:688 selective removal 672:688 selective removal of acetyl groups 672:705 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 2 7 theme temporary 457:465 arg1 protection 467:476 a temporary protection 455:476 a temporary protection of the diol system 455:495 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 7 theme temporary 457:465 arg1 glycosylation 295:307 highly regioselective glycosylation 273:307 highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 273:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 1 8 link -linked 139:145 arg1 → 135:135 1 → 3 133:137 1 → 3 133:137 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 1 8 link -linked 139:145 arg1 D-glucooligosaccharides 147:169 linear β-(1 → 3)-linked D-glucooligosaccharides 123:169 linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 123:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 3 9 theme steps 546:550 arg1 Iteration 498:506 Iteration 498:506 Iteration of the deprotection and glycosylation steps 498:550 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 2 10 theme chloroacetyl 416:427 arg1 groups 429:434 chloroacetyl groups 416:434 chloroacetyl groups 416:434 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 4 11 theme brief 755:759 arg1 treatment 761:769 a brief treatment 753:769 a brief treatment with a large excess of hydrazine hydrate 753:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 3 12 theme title 579:583 arg1 oligoglucosides 585:599 the title oligoglucosides 575:599 the title oligoglucosides differing in length by two monosaccharide units 575:647 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 4 13 from procedure 658:666 arg1 presence 714:721 the presence 710:721 the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate 710:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 1 14 theme -linked 139:145 arg1 → 135:135 1 → 3 133:137 1 → 3 133:137 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 1 14 theme -linked 139:145 arg1 D-glucooligosaccharides 147:169 linear β-(1 → 3)-linked D-glucooligosaccharides 123:169 linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 123:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 0 15 theme glycosides 27:36 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3) 0:56 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 2 16 theme disaccharide 375:386 arg1 donor 402:406 a disaccharide thioglycoside donor 373:406 a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 373:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 1 17 theme 3-Aminopropyl 83:95 arg1 glycosides 97:106 3-Aminopropyl glycosides 83:106 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 83:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 1 18 theme D-glucooligosaccharides 147:169 arg1 series 113:118 a series 111:118 a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 111:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 0 19 theme 3-aminopropyl 13:25 arg1 glycosides 27:36 3-aminopropyl glycosides 13:36 3-aminopropyl glycosides 13:36 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 3 20 theme glycosylation 532:544 arg1 steps 546:550 the deprotection and glycosylation steps 511:550 steps 546:550 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 1 21 theme linear 123:128 arg1 → 135:135 1 → 3 133:137 1 → 3 133:137 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 1 21 theme linear 123:128 arg1 D-glucooligosaccharides 147:169 linear β-(1 → 3)-linked D-glucooligosaccharides 123:169 linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 123:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 0 22 theme linear 41:46 arg1 β- 48:49 linear β- 41:49 linear β-(1 → 3) 41:56 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 4 23 with treatment 761:769 arg1 excess 784:789 a large excess 776:789 a large excess of hydrazine hydrate 776:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 4 24 theme groups 700:705 arg1 removal 682:688 selective removal 672:688 selective removal of acetyl groups 672:705 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 2 25 with acceptors 358:366 arg1 donor 402:406 a disaccharide thioglycoside donor 373:406 a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 373:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 0 26 dep β- 48:49 arg1 3 55:55 3 55:55 3 55:55 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 0 26 dep β- 48:49 arg1 1 51:51 1 51:51 1 51:51 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 4 27 theme novel 652:656 arg1 procedure 658:666 A novel procedure 650:666 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate 650:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 1 28 dep 13 192:193 arg1 to 189:190 to 189:190 to 189:190 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 3 29 theme oligoglucosides 585:599 arg1 series 565:570 the series 561:570 the series of the title oligoglucosides differing in length by two monosaccharide units 561:647 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 2 30 theme acceptors 358:366 arg1 protection 467:476 a temporary protection 455:476 a temporary protection of the diol system 455:495 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 30 theme acceptors 358:366 arg1 glycosylation 295:307 highly regioselective glycosylation 273:307 highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 273:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 0 31 theme β- 48:49 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3) 0:56 Synthesis of 3-aminopropyl glycosides of linear β-(1 → 3)-D-glucooligosaccharides. 26595660 2 32 theme glycosyl 349:356 arg1 acceptors 358:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors 312:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 312:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 3 33 theme monosaccharide 628:641 arg1 units 643:647 two monosaccharide units 624:647 two monosaccharide units 624:647 Iteration of the deprotection and glycosylation steps afforded the series of the title oligoglucosides differing in length by two monosaccharide units. 26595660 4 34 theme acetyl 693:698 arg1 groups 700:705 acetyl groups 693:705 acetyl groups 693:705 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 1 35 theme series 113:118 arg1 glycosides 97:106 3-Aminopropyl glycosides 83:106 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 83:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 2 36 theme regioselective 280:293 arg1 protection 467:476 a temporary protection 455:476 a temporary protection of the diol system 455:495 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 36 theme regioselective 280:293 arg1 glycosylation 295:307 highly regioselective glycosylation 273:307 highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 273:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 4 37 theme large 778:782 arg1 excess 784:789 a large excess 776:789 a large excess of hydrazine hydrate 776:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 2 38 theme 4,6-O-benzylidene-protected 312:338 arg1 acceptors 358:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors 312:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 312:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 39 theme 2,3-diol 340:347 arg1 acceptors 358:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors 312:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 312:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 2 40 gly glycosylation 295:307 arg1 acceptors 358:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors 312:366 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 312:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 4 41 theme hydrate 804:810 arg1 excess 784:789 a large excess 776:789 a large excess of hydrazine hydrate 776:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 2 42 theme thioglycoside 388:400 arg1 donor 402:406 a disaccharide thioglycoside donor 373:406 a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' 373:450 The synthetic scheme featured highly regioselective glycosylation of 4,6-O-benzylidene-protected 2,3-diol glycosyl acceptors with a disaccharide thioglycoside donor bearing chloroacetyl groups at O-2' and -3' as a temporary protection of the diol system. 26595660 1 43 theme monosaccharide 195:208 arg1 units 210:214 3 to 13 monosaccharide units 187:214 3 to 13 monosaccharide units 187:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 4 44 theme hydrazine 794:802 arg1 hydrate 804:810 hydrazine hydrate 794:810 hydrazine hydrate 794:810 A novel procedure for selective removal of acetyl groups in the presence of benzoyl ones consisting in a brief treatment with a large excess of hydrazine hydrate has been proposed. 26595660 1 45 theme β- 130:131 arg1 → 135:135 1 → 3 133:137 1 → 3 133:137 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 26595660 1 45 theme β- 130:131 arg1 D-glucooligosaccharides 147:169 linear β-(1 → 3)-linked D-glucooligosaccharides 123:169 linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units 123:214 3-Aminopropyl glycosides of a series of linear β-(1 → 3)-linked D-glucooligosaccharides containing from 3 to 13 monosaccharide units were efficiently prepared. 25638265 0 0 theme liquid 103:108 arg1 spectrometry 154:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 3 1 theme quantitative 733:744 arg1 analysis 746:753 the quantitative analysis 729:753 the quantitative analysis in combined samples 729:773 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 1 2 theme reductive 214:222 arg1 GRIL 242:245 GRIL 242:245 GRIL 242:245 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 1 2 theme reductive 214:222 arg1 labeling 232:239 Glycan reductive isotope labeling 207:239 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent 207:295 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 3 3 dep accuracy 703:710 arg1 the 690:692 the 690:692 the 690:692 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 0 4 theme performance 91:101 arg1 spectrometry 154:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 5 5 theme including 1197:1205 arg1 amounts 1175:1181 relative amounts 1166:1181 relative amounts of N-glycans (including isomers) 1166:1214 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 5 theme including 1197:1205 arg1 N-glycans 1186:1194 N-glycans 1186:1194 N-glycans 1186:1194 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 5 theme including 1197:1205 arg1 isomers 1207:1213 including isomers 1197:1213 including isomers 1197:1213 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 1 6 theme isotope 224:230 arg1 GRIL 242:245 GRIL 242:245 GRIL 242:245 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 1 6 theme isotope 224:230 arg1 labeling 232:239 Glycan reductive isotope labeling 207:239 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent 207:295 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 3 7 from accuracy 703:710 arg1 samples 767:773 combined samples 758:773 combined samples 758:773 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 3 8 theme identical 898:906 arg1 proteins 915:922 identical target proteins 898:922 identical target proteins 898:922 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 0 9 theme ionization-mass 138:152 arg1 spectrometry 154:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 6 10 with combination 1316:1326 arg1 PGC-ESI-TOF-MS 1333:1346 PGC-ESI-TOF-MS 1333:1346 PGC-ESI-TOF-MS 1333:1346 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 1 11 theme labeling 280:287 arg1 reagent 289:295 labeling reagent 280:295 labeling reagent 280:295 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 0 12 theme chromatography-electrospray 110:136 arg1 spectrometry 154:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 3 13 theme combined 758:765 arg1 samples 767:773 combined samples 758:773 combined samples 758:773 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 6 14 theme Chinese 1431:1437 arg1 cells 1453:1457 Chinese hamster ovary cells 1431:1457 Chinese hamster ovary cells (CHO) 1431:1463 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 14 theme Chinese 1431:1437 arg1 CHO 1460:1462 CHO 1460:1462 CHO 1460:1462 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 15 theme ovary 1447:1451 arg1 cells 1453:1457 Chinese hamster ovary cells 1431:1457 Chinese hamster ovary cells (CHO) 1431:1463 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 15 theme ovary 1447:1451 arg1 CHO 1460:1462 CHO 1460:1462 CHO 1460:1462 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 5 16 theme relative 1166:1173 arg1 amounts 1175:1181 relative amounts 1166:1181 relative amounts of N-glycans (including isomers) 1166:1214 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 16 theme relative 1166:1173 arg1 N-glycans 1186:1194 N-glycans 1186:1194 N-glycans 1186:1194 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 16 theme relative 1166:1173 arg1 isomers 1207:1213 including isomers 1197:1213 including isomers 1197:1213 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 6 17 gly glycosylation 1374:1386 arg1 antibodies 1408:1417 human monoclonal antibodies 1391:1417 human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines 1391:1488 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 18 theme GRIL 1308:1311 arg1 usefulness 1294:1303 usefulness 1294:1303 usefulness 1294:1303 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 18 theme GRIL 1308:1311 arg1 potential 1280:1288 analytical potential 1269:1288 analytical potential 1269:1288 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 19 theme hamster 1439:1445 arg1 cells 1453:1457 Chinese hamster ovary cells 1431:1457 Chinese hamster ovary cells (CHO) 1431:1463 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 19 theme hamster 1439:1445 arg1 CHO 1460:1462 CHO 1460:1462 CHO 1460:1462 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 3 20 theme analysis 746:753 arg1 accuracy 703:710 improved accuracy 694:710 improved accuracy 694:710 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 2 21 theme flight 494:499 arg1 analyzer 501:508 flight analyzer 494:508 flight analyzer 494:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 22 theme mass 463:466 arg1 spectrometry 468:479 electrospray mass spectrometry 450:479 electrospray mass spectrometry 450:479 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 0 23 theme isomer-specific 13:27 arg1 N-glycan 29:36 Quantitative isomer-specific N-glycan 0:36 Quantitative isomer-specific N-glycan 0:36 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 2 24 theme analyzer 501:508 arg1 time 486:489 time 486:489 time of flight analyzer 486:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 6 25 theme hybridoma 1469:1477 arg1 lines 1484:1488 hybridoma cell lines 1469:1488 hybridoma cell lines 1469:1488 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 3 26 theme main 598:601 arg1 benefit 603:609 The main benefit 594:609 The main benefit of using stable isotope-coding in the context of comparative glycomics 594:680 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 6 27 theme human 1391:1395 arg1 antibodies 1408:1417 human monoclonal antibodies 1391:1417 human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines 1391:1488 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 2 28 theme stationary 398:407 arg1 phase 409:413 stationary phase 398:413 stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 398:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 0 29 theme Quantitative 0:11 arg1 N-glycan 29:36 Quantitative isomer-specific N-glycan 0:36 Quantitative isomer-specific N-glycan 0:36 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 5 30 theme developed 1123:1131 arg1 capable 1143:1149 capable 1143:1149 capable 1143:1149 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 30 theme developed 1123:1131 arg1 method 1133:1138 the developed method 1119:1138 the developed method 1119:1138 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 2 31 theme electrospray 450:461 arg1 spectrometry 468:479 electrospray mass spectrometry 450:479 electrospray mass spectrometry 450:479 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 6 32 dep potential 1280:1288 arg1 The 1265:1267 The 1265:1267 The 1265:1267 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 0 33 theme carbon 182:187 arg1 phase 200:204 graphitic carbon stationary phase 172:204 graphitic carbon stationary phase 172:204 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 4 34 gly glycoproteins 1087:1099 arg1 glycoproteins 1087:1099 test glycoproteins 1082:1099 test glycoproteins 1082:1099 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 3 35 from analysis 746:753 arg1 samples 767:773 combined samples 758:773 combined samples 758:773 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 3 36 from samples 767:773 arg1 accuracy 703:710 improved accuracy 694:710 improved accuracy 694:710 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 5 37 theme single 1245:1250 arg1 run 1260:1262 one single HPLC-MS run 1241:1262 one single HPLC-MS run 1241:1262 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 0 38 theme graphitic 172:180 arg1 phase 200:204 graphitic carbon stationary phase 172:204 graphitic carbon stationary phase 172:204 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 0 39 theme isotope 59:65 arg1 labeling 73:80 isotope coded labeling 59:80 isotope coded labeling 59:80 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 5 40 theme HPLC-MS 1252:1258 arg1 run 1260:1262 one single HPLC-MS run 1241:1262 one single HPLC-MS run 1241:1262 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 3 41 theme stable 620:625 arg1 isotope-coding 627:640 stable isotope-coding 620:640 stable isotope-coding 620:640 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 2 42 attach released 565:572 arg2 N-glycans 555:563 labeled N-glycans 547:563 labeled N-glycans released from glycoproteins 547:591 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 42 attach released 565:572 arg1 glycoproteins 579:591 glycoproteins 579:591 glycoproteins 579:591 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 43 theme scale 428:432 arg1 HPLC 434:437 capillary scale HPLC 418:437 capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 418:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 4 44 theme detection/quantification 1041:1064 arg1 linearity 1018:1026 linearity 1018:1026 linearity 1018:1026 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 44 theme detection/quantification 1041:1064 arg1 accuracy 1008:1015 accuracy 1008:1015 accuracy 1008:1015 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 44 theme detection/quantification 1041:1064 arg1 reproducibility 991:1005 reproducibility 991:1005 reproducibility 991:1005 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 44 theme detection/quantification 1041:1064 arg1 parameters 979:988 mobile phase parameters 966:988 mobile phase parameters 966:988 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 44 theme detection/quantification 1041:1064 arg1 limit 1032:1036 limit 1032:1036 limit 1032:1036 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 0 45 theme stationary 189:198 arg1 phase 200:204 graphitic carbon stationary phase 172:204 graphitic carbon stationary phase 172:204 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 3 46 theme comparative 660:670 arg1 glycomics 672:680 comparative glycomics 660:680 comparative glycomics 660:680 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 2 47 theme N-glycans 555:563 arg1 determination 530:542 the determination 526:542 the determination of labeled N-glycans released from glycoproteins 526:591 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 48 theme capillary 418:426 arg1 HPLC 434:437 capillary scale HPLC 418:437 capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 418:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 4 49 theme test 1082:1085 arg1 glycoproteins 1087:1099 test glycoproteins 1082:1099 test glycoproteins 1082:1099 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 2 50 theme labeled 547:553 arg1 N-glycans 555:563 labeled N-glycans 547:563 labeled N-glycans released from glycoproteins 547:591 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 6 51 theme monoclonal 1397:1406 arg1 antibodies 1408:1417 human monoclonal antibodies 1391:1417 human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines 1391:1488 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 3 52 theme glycomics 672:680 arg1 context 649:655 the context 645:655 the context of comparative glycomics 645:680 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 2 53 theme graphitized 370:380 arg1 PGC 390:392 PGC 390:392 PGC 390:392 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 53 theme graphitized 370:380 arg1 carbon 382:387 Porous graphitized carbon 363:387 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 363:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 0 54 theme coded 67:71 arg1 labeling 73:80 isotope coded labeling 59:80 isotope coded labeling 59:80 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 1 55 theme quantitative 325:336 arg1 fingerprinting 347:360 quantitative N-glycan fingerprinting 325:360 quantitative N-glycan fingerprinting 325:360 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 5 56 theme N-glycans 1186:1194 arg1 amounts 1175:1181 relative amounts 1166:1181 relative amounts of N-glycans (including isomers) 1166:1214 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 56 theme N-glycans 1186:1194 arg1 N-glycans 1186:1194 N-glycans 1186:1194 N-glycans 1186:1194 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 5 56 theme N-glycans 1186:1194 arg1 isomers 1207:1213 including isomers 1197:1213 including isomers 1197:1213 It is shown that the developed method is capable of determining relative amounts of N-glycans (including isomers) comparing two samples in one single HPLC-MS run. 25638265 2 57 theme Porous 363:368 arg1 PGC 390:392 PGC 390:392 PGC 390:392 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 2 57 theme Porous 363:368 arg1 carbon 382:387 Porous graphitized carbon 363:387 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 363:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 1 58 theme N-glycan 338:345 arg1 fingerprinting 347:360 quantitative N-glycan fingerprinting 325:360 quantitative N-glycan fingerprinting 325:360 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 4 59 theme phase 973:977 arg1 linearity 1018:1026 linearity 1018:1026 linearity 1018:1026 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 59 theme phase 973:977 arg1 accuracy 1008:1015 accuracy 1008:1015 accuracy 1008:1015 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 59 theme phase 973:977 arg1 reproducibility 991:1005 reproducibility 991:1005 reproducibility 991:1005 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 59 theme phase 973:977 arg1 limit 1032:1036 limit 1032:1036 limit 1032:1036 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 59 theme phase 973:977 arg1 parameters 979:988 mobile phase parameters 966:988 mobile phase parameters 966:988 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 6 60 theme analytical 1269:1278 arg1 potential 1280:1288 analytical potential 1269:1288 analytical potential 1269:1288 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 2 61 from phase 409:413 arg1 HPLC 434:437 capillary scale HPLC 418:437 capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer 418:508 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 6 62 from potential 1280:1288 arg1 combination 1316:1326 combination 1316:1326 combination with PGC-ESI-TOF-MS 1316:1346 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 1 63 theme C6-/ 258:261 arg1 C6-aniline 266:275 (12)C6-/(13)C6-aniline 254:275 (12)C6-/(13)C6-aniline 254:275 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 4 64 theme mobile 966:971 arg1 linearity 1018:1026 linearity 1018:1026 linearity 1018:1026 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 64 theme mobile 966:971 arg1 accuracy 1008:1015 accuracy 1008:1015 accuracy 1008:1015 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 64 theme mobile 966:971 arg1 reproducibility 991:1005 reproducibility 991:1005 reproducibility 991:1005 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 64 theme mobile 966:971 arg1 limit 1032:1036 limit 1032:1036 limit 1032:1036 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 4 64 theme mobile 966:971 arg1 parameters 979:988 mobile phase parameters 966:988 mobile phase parameters 966:988 The method was validated with respect to mobile phase parameters, reproducibility, accuracy, linearity and limit of detection/quantification (LOD/LOQ) using test glycoproteins. 25638265 1 65 theme Glycan 207:212 arg1 GRIL 242:245 GRIL 242:245 GRIL 242:245 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 1 65 theme Glycan 207:212 arg1 labeling 232:239 Glycan reductive isotope labeling 207:239 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent 207:295 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 3 66 theme target 908:913 arg1 proteins 915:922 identical target proteins 898:922 identical target proteins 898:922 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 0 67 theme high 86:89 arg1 spectrometry 154:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 high performance liquid chromatography-electrospray ionization-mass spectrometry 86:165 Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography-electrospray ionization-mass spectrometry with graphitic carbon stationary phase. 25638265 3 68 theme improved 694:701 arg1 accuracy 703:710 improved accuracy 694:710 improved accuracy 694:710 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 6 69 theme cell 1479:1482 arg1 lines 1484:1488 hybridoma cell lines 1469:1488 hybridoma cell lines 1469:1488 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 6 70 from usefulness 1294:1303 arg1 combination 1316:1326 combination 1316:1326 combination with PGC-ESI-TOF-MS 1316:1346 The analytical potential and usefulness of GRIL in combination with PGC-ESI-TOF-MS is demonstrated comparing glycosylation in human monoclonal antibodies produced in Chinese hamster ovary cells (CHO) and hybridoma cell lines. 25638265 3 71 theme incomplete 834:843 arg1 release 855:861 structure-dependent incomplete enzymatic release 814:861 structure-dependent incomplete enzymatic release of oligosaccharides 814:881 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 3 72 theme oligosaccharides 866:881 arg1 release 855:861 structure-dependent incomplete enzymatic release 814:861 structure-dependent incomplete enzymatic release of oligosaccharides 814:881 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 1 73 theme fingerprinting 347:360 arg1 aim 318:320 the aim 314:320 the aim of quantitative N-glycan fingerprinting 314:360 Glycan reductive isotope labeling (GRIL) using (12)C6-/(13)C6-aniline as labeling reagent is reported with the aim of quantitative N-glycan fingerprinting. 25638265 3 74 theme enzymatic 845:853 arg1 release 855:861 structure-dependent incomplete enzymatic release 814:861 structure-dependent incomplete enzymatic release of oligosaccharides 814:881 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 25638265 2 75 gly glycoproteins 579:591 arg1 glycoproteins 579:591 glycoproteins 579:591 glycoproteins 579:591 Porous graphitized carbon (PGC) as stationary phase in capillary scale HPLC coupled to electrospray mass spectrometry with time of flight analyzer was applied for the determination of labeled N-glycans released from glycoproteins. 25638265 3 76 theme structure-dependent 814:832 arg1 release 855:861 structure-dependent incomplete enzymatic release 814:861 structure-dependent incomplete enzymatic release of oligosaccharides 814:881 The main benefit of using stable isotope-coding in the context of comparative glycomics lies in the improved accuracy and precision of the quantitative analysis in combined samples and in the potential of correcting for structure-dependent incomplete enzymatic release of oligosaccharides when comparing identical target proteins. 28318087 1 0 dep concise 105:111 arg1 diversity-oriented 114:131 diversity-oriented 114:131 diversity-oriented 114:131 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 2 1 from application 387:397 arg1 reactions 416:424 glycosylation reactions 402:424 glycosylation reactions 402:424 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 0 2 from 3-Aminodeoxypyranoses 0:20 arg1 Glycosylation 25:37 Glycosylation 25:37 Glycosylation 25:37 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 0 3 from Assembly 73:80 arg1 Oligosaccharides 85:100 Oligosaccharides 85:100 Oligosaccharides 85:100 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 1 4 theme 3-amino- 183:190 arg1 synthesis 150:158 the synthesis 146:158 the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars 146:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 1 5 theme 3-nitro-2,3,6-trideoxypyranose 196:225 arg1 derivatives 227:237 naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives 163:237 derivatives 227:237 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 0 6 from Synthesis 59:67 arg1 Oligosaccharides 85:100 Oligosaccharides 85:100 Oligosaccharides 85:100 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 1 7 theme derivatives 227:237 arg1 synthesis 150:158 the synthesis 146:158 the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars 146:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 1 8 theme concise 105:111 arg1 approach 133:140 A concise, diversity-oriented approach 103:140 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars 103:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 3 9 theme successful 452:461 arg1 synthesis 463:471 the successful synthesis 448:471 the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors 448:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 2 10 theme donors 370:375 arg1 synthesis 333:341 the synthesis 329:341 the synthesis of various 3-aminoglycosyl donors 329:375 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 2 10 theme donors 370:375 arg1 application 387:397 their application 381:397 their application in glycosylation reactions 381:424 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 1 11 theme analogues 243:251 arg1 synthesis 150:158 the synthesis 146:158 the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars 146:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 2 12 theme 3-aminoglycosyl 354:368 arg1 donors 370:375 various 3-aminoglycosyl donors 346:375 various 3-aminoglycosyl donors 346:375 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 3 13 theme ortho-alkynylbenzoate 550:570 arg1 donors 572:577 ortho-alkynylbenzoate donors 550:577 ortho-alkynylbenzoate donors 550:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 2 14 theme various 346:352 arg1 donors 370:375 various 3-aminoglycosyl donors 346:375 various 3-aminoglycosyl donors 346:375 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 1 15 theme simple 266:271 arg1 sugars 273:278 simple sugars 266:278 simple sugars 266:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 3 16 theme tetrasaccharide 478:492 arg1 synthesis 463:471 the successful synthesis 448:471 the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors 448:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 0 17 theme Diversity-Oriented 40:57 arg1 Synthesis 59:67 Synthesis 59:67 Synthesis 59:67 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 0 17 theme Diversity-Oriented 40:57 arg1 3-Aminodeoxypyranoses 0:20 3-Aminodeoxypyranoses 0:20 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 0:101 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 3 18 theme different 510:518 arg1 components 533:542 four different 3-aminosugar components 505:542 four different 3-aminosugar components using ortho-alkynylbenzoate donors 505:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 1 19 from sugars 273:278 arg1 synthesis 150:158 the synthesis 146:158 the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars 146:278 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 28318087 2 20 theme glycosylation 402:414 arg1 reactions 416:424 glycosylation reactions 402:424 glycosylation reactions 402:424 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 3 21 contain containing 494:503 arg2 components 533:542 four different 3-aminosugar components 505:542 four different 3-aminosugar components using ortho-alkynylbenzoate donors 505:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 3 21 contain containing 494:503 arg1 tetrasaccharide 478:492 a tetrasaccharide 476:492 a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors 476:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 3 22 theme 3-aminosugar 520:531 arg1 components 533:542 four different 3-aminosugar components 505:542 four different 3-aminosugar components using ortho-alkynylbenzoate donors 505:577 These studies led to the successful synthesis of a tetrasaccharide containing four different 3-aminosugar components using ortho-alkynylbenzoate donors. 28318087 2 23 from synthesis 333:341 arg1 reactions 416:424 glycosylation reactions 402:424 glycosylation reactions 402:424 In addition, we investigated the synthesis of various 3-aminoglycosyl donors and their application in glycosylation reactions. 28318087 0 24 dep 3-Aminodeoxypyranoses 0:20 arg1 Synthesis 59:67 Synthesis 59:67 Synthesis 59:67 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 0 24 dep 3-Aminodeoxypyranoses 0:20 arg1 3-Aminodeoxypyranoses 0:20 3-Aminodeoxypyranoses 0:20 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 0:101 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 0 24 dep 3-Aminodeoxypyranoses 0:20 arg1 Assembly 73:80 Assembly 73:80 Assembly 73:80 3-Aminodeoxypyranoses in Glycosylation: Diversity-Oriented Synthesis and Assembly in Oligosaccharides. 28318087 1 25 theme occurring 173:181 arg1 3-amino- 183:190 naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives 163:237 3-amino- 183:190 A concise, diversity-oriented approach for the synthesis of naturally occurring 3-amino- and 3-nitro-2,3,6-trideoxypyranose derivatives and analogues thereof from simple sugars has been developed. 29156420 2 0 theme orthogonal 426:435 arg1 coupling 437:444 orthogonal coupling 426:444 orthogonal coupling between thioglycosides of L-rhamnose 426:481 This synthesis was achieved by iterative α-glycosylation using disaccharide building blocks and through orthogonal coupling between thioglycosides of L-rhamnose. 29156420 0 1 theme iterative 122:130 arg1 α-glycosylation 132:146 iterative α-glycosylation 122:146 iterative α-glycosylation using disaccharide building blocks 122:181 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 2 theme sulfation 862:870 arg1 patterns 872:879 diverse sulfation patterns 854:879 diverse sulfation patterns 854:879 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 3 theme different 666:674 arg1 groups 698:703 different orthogonal protecting groups 666:703 different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) 666:769 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 0 4 dep -linkages 78:86 arg1 1→2 68:70 1→2 68:70 1→2 68:70 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 4 dep -linkages 78:86 arg1 1→3 74:76 1→3 74:76 1→3 74:76 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 5 theme structures 838:847 arg1 range 821:825 a range 819:825 a range of rhamnan structures with diverse sulfation patterns 819:879 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 1 6 theme rhamnan 213:219 arg1 oligosaccharides 221:236 rhamnan oligosaccharides 213:236 rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages 213:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 1 7 theme repeating 282:290 arg1 -linkages 299:307 repeating α-(1→3)-linkages 282:307 repeating α-(1→3)-linkages 282:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 2 8 theme L-rhamnose 472:481 arg1 thioglycosides 454:467 thioglycosides 454:467 thioglycosides of L-rhamnose 454:481 This synthesis was achieved by iterative α-glycosylation using disaccharide building blocks and through orthogonal coupling between thioglycosides of L-rhamnose. 29156420 1 9 theme α- 292:293 arg1 -linkages 299:307 repeating α-(1→3)-linkages 282:307 repeating α-(1→3)-linkages 282:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 3 10 theme detailed 503:510 arg1 relationship 531:542 the detailed structure-activity relationship 499:542 the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 499:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 0 11 theme building 167:174 arg1 blocks 176:181 disaccharide building blocks 154:181 disaccharide building blocks 154:181 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 12 dep Monostroma 568:577 arg1 nitidum 579:585 Monostroma nitidum 568:585 Monostroma nitidum 568:585 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 1 13 theme 1→3 295:297 arg1 -linkages 299:307 repeating α-(1→3)-linkages 282:307 repeating α-(1→3)-linkages 282:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 0 14 theme disaccharide 154:165 arg1 blocks 176:181 disaccharide building blocks 154:181 disaccharide building blocks 154:181 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 15 theme concise 2:8 arg1 synthesis 10:18 A concise synthesis 0:18 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3) 0:108 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 16 theme orthogonal 676:685 arg1 groups 698:703 different orthogonal protecting groups 666:703 different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) 666:769 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 1 17 theme alternating 243:253 arg1 -linkages 268:276 alternating α-(1→2)/(1→3)-linkages 243:276 alternating α-(1→2)/(1→3)-linkages 243:276 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 3 18 theme rhamnan 547:553 arg1 sulfate 555:561 rhamnan sulfate 547:561 rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 547:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 2 19 theme iterative 353:361 arg1 α-glycosylation 363:377 iterative α-glycosylation 353:377 iterative α-glycosylation using disaccharide building blocks 353:412 This synthesis was achieved by iterative α-glycosylation using disaccharide building blocks and through orthogonal coupling between thioglycosides of L-rhamnose. 29156420 3 20 theme structure-activity 512:529 arg1 relationship 531:542 the detailed structure-activity relationship 499:542 the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 499:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 21 from Monostroma 568:577 arg1 relationship 531:542 the detailed structure-activity relationship 499:542 the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 499:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 21 from Monostroma 568:577 arg1 sulfate 555:561 rhamnan sulfate 547:561 rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 547:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 0 22 theme oligosaccharides 31:46 arg1 synthesis 10:18 A concise synthesis 0:18 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3) 0:108 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 1 23 theme 1→2 258:260 arg1 -linkages 268:276 alternating α-(1→2)/(1→3)-linkages 243:276 alternating α-(1→2)/(1→3)-linkages 243:276 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 0 24 theme rhamnan 23:29 arg1 oligosaccharides 31:46 rhamnan oligosaccharides 23:46 rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3) 23:108 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 25 theme repeating 92:100 arg1 1→3 105:107 1→3 105:107 1→3 105:107 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 25 theme repeating 92:100 arg1 α- 102:103 repeating α- 92:103 repeating α-(1→3) 92:108 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 1 26 with oligosaccharides 221:236 arg1 -linkages 299:307 repeating α-(1→3)-linkages 282:307 repeating α-(1→3)-linkages 282:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 1 26 with oligosaccharides 221:236 arg1 -linkages 268:276 alternating α-(1→2)/(1→3)-linkages 243:276 alternating α-(1→2)/(1→3)-linkages 243:276 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 2 27 theme building 398:405 arg1 blocks 407:412 disaccharide building blocks 385:412 disaccharide building blocks 385:412 This synthesis was achieved by iterative α-glycosylation using disaccharide building blocks and through orthogonal coupling between thioglycosides of L-rhamnose. 29156420 0 28 theme α- 65:66 arg1 -linkages 78:86 alternating α-(1→2)/(1→3)-linkages 53:86 alternating α-(1→2)/(1→3)-linkages 53:86 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 29 theme sulfate 555:561 arg1 relationship 531:542 the detailed structure-activity relationship 499:542 the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2 499:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 2 30 theme disaccharide 385:396 arg1 blocks 407:412 disaccharide building blocks 385:412 disaccharide building blocks 385:412 This synthesis was achieved by iterative α-glycosylation using disaccharide building blocks and through orthogonal coupling between thioglycosides of L-rhamnose. 29156420 3 31 with structures 838:847 arg1 patterns 872:879 diverse sulfation patterns 854:879 diverse sulfation patterns 854:879 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 32 dep benzoyl 712:718 arg1 i.e. 706:709 i.e. 706:709 i.e. 706:709 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 33 theme simplex 602:608 arg1 type 616:619 herpes simplex virus type 2 595:621 herpes simplex virus type 2 595:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 34 theme rhamnan 830:836 arg1 structures 838:847 rhamnan structures 830:847 rhamnan structures with diverse sulfation patterns 830:879 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 35 theme synthesized 628:638 arg1 oligosaccharides 640:655 the synthesized oligosaccharides 624:655 the synthesized oligosaccharides 624:655 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 35 theme synthesized 628:638 arg1 suitable 790:797 suitable 790:797 suitable 790:797 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 36 theme virus 610:614 arg1 type 616:619 herpes simplex virus type 2 595:621 herpes simplex virus type 2 595:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 1 37 theme concise 186:192 arg1 route 204:208 A concise synthetic route 184:208 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages 184:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 3 38 from relationship 531:542 arg1 Monostroma 568:577 Monostroma 568:577 Monostroma 568:577 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 1 39 theme synthetic 194:202 arg1 route 204:208 A concise synthetic route 184:208 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages 184:307 A concise synthetic route to rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages is reported. 29156420 0 40 theme alternating 53:63 arg1 -linkages 78:86 alternating α-(1→2)/(1→3)-linkages 53:86 alternating α-(1→2)/(1→3)-linkages 53:86 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 41 theme protecting 687:696 arg1 groups 698:703 different orthogonal protecting groups 666:703 different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) 666:769 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 3 42 theme herpes 595:600 arg1 type 616:619 herpes simplex virus type 2 595:621 herpes simplex virus type 2 595:621 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 29156420 0 43 with oligosaccharides 31:46 arg1 1→3 105:107 1→3 105:107 1→3 105:107 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 43 with oligosaccharides 31:46 arg1 -linkages 78:86 alternating α-(1→2)/(1→3)-linkages 53:86 alternating α-(1→2)/(1→3)-linkages 53:86 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 0 43 with oligosaccharides 31:46 arg1 α- 102:103 repeating α- 92:103 repeating α-(1→3) 92:108 A concise synthesis of rhamnan oligosaccharides with alternating α-(1→2)/(1→3)-linkages and repeating α-(1→3)-linkages by iterative α-glycosylation using disaccharide building blocks. 29156420 3 44 theme diverse 854:860 arg1 patterns 872:879 diverse sulfation patterns 854:879 diverse sulfation patterns 854:879 To investigate the detailed structure-activity relationship of rhamnan sulfate from Monostroma nitidum against herpes simplex virus type 2, the synthesized oligosaccharides, bearing different orthogonal protecting groups (i.e., benzoyl, benzyl, 2-naphthylmethyl, and/or p-methoxybenzyl) are expected to be suitable for conversion into a range of rhamnan structures with diverse sulfation patterns. 28375048 4 0 theme mammalian 1178:1186 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 0 theme mammalian 1178:1186 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 1 1 theme therapeutics 314:325 arg1 properties 258:267 the physiochemical properties 239:267 the physiochemical properties 239:267 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 1 1 theme therapeutics 314:325 arg1 efficacy 277:284 the efficacy 273:284 the efficacy of recombinant glycoprotein therapeutics 273:325 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 2 theme culture 1193:1199 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 2 theme culture 1193:1199 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 8 3 theme N-glycan 1832:1839 arg1 expansion 1815:1823 an expansion 1812:1823 an expansion of the N-glycan code in mammalian expressed glycoproteins 1812:1881 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 2 4 theme immunoglobulins 351:365 arg1 Glycosylation 328:340 Glycosylation 328:340 Glycosylation of human immunoglobulins 328:365 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 5 5 theme adaptation 1349:1358 arg1 strategies 1360:1369 no cell line engineering or cellular adaptation strategies 1312:1369 no cell line engineering or cellular adaptation strategies 1312:1369 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 4 6 theme immunoglobulin 953:966 arg1 preparations 968:979 recombinant human immunoglobulin preparations 935:979 recombinant human immunoglobulin preparations 935:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 7 theme pharmacokinetic 809:823 arg1 profiles 825:832 less favorable pharmacokinetic profiles 794:832 less favorable pharmacokinetic profiles 794:832 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 5 8 theme engineering 1325:1335 arg1 strategies 1360:1369 no cell line engineering or cellular adaptation strategies 1312:1369 no cell line engineering or cellular adaptation strategies 1312:1369 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 7 9 theme aforementioned 1608:1621 arg1 effects 1623:1629 These aforementioned effects 1602:1629 These aforementioned effects 1602:1629 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 2 10 gly Glycosylation 328:340 arg1 immunoglobulins 351:365 human immunoglobulins 345:365 human immunoglobulins 345:365 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 6 11 theme Arabinosylated 1386:1399 arg1 IgGs 1413:1416 Arabinosylated recombinant IgGs 1386:1416 Arabinosylated recombinant IgGs 1386:1416 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 2 12 theme well-understood 388:402 arg1 pathway 414:420 a reasonably well-understood metabolic pathway 375:420 a reasonably well-understood metabolic pathway 375:420 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 1 13 theme physiochemical 243:256 arg1 properties 258:267 the physiochemical properties 239:267 the physiochemical properties 239:267 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 1 14 gly glycoproteins 175:187 arg1 glycoproteins 175:187 recombinant glycoproteins 163:187 recombinant glycoproteins 163:187 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 15 theme approximate 1007:1017 arg1 replacement 1024:1034 an approximate 100% replacement 1004:1034 an approximate 100% replacement of fucosylation with arabinosylation 1004:1071 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 16 theme cell-mediated 660:672 arg1 cytotoxicity 674:685 antibody-dependent cell-mediated cytotoxicity 641:685 antibody-dependent cell-mediated cytotoxicity 641:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 1 17 theme recombinant 163:173 arg1 glycoproteins 175:187 recombinant glycoproteins 163:187 recombinant glycoproteins 163:187 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 18 theme % 1022:1022 arg1 replacement 1024:1034 an approximate 100% replacement 1004:1034 an approximate 100% replacement of fucosylation with arabinosylation 1004:1071 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 6 19 theme high 1452:1455 arg1 glycans 1465:1471 high mannose glycans 1452:1471 high mannose glycans 1452:1471 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 4 20 theme cell 1098:1101 arg1 culture 1103:1109 Chinese hamster ovary cell culture 1076:1109 Chinese hamster ovary cell culture 1076:1109 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 8 21 dep N-glycan 1832:1839 arg1 code 1841:1844 code 1841:1844 code 1841:1844 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 3 22 theme high 692:695 arg1 levels 705:710 high mannose levels 692:710 high mannose levels 692:710 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 7 23 theme bispecific 1758:1767 arg1 DVD-Ig 1769:1774 a bispecific DVD-Ig 1756:1774 a bispecific DVD-Ig 1756:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 8 24 from expansion 1815:1823 arg1 glycoproteins 1869:1881 mammalian expressed glycoproteins 1849:1881 mammalian expressed glycoproteins 1849:1881 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 2 25 link serine/threonine-linked 495:517 arg1 glycans 530:536 asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 462:536 glycans 530:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 4 26 attach presence 899:906 arg1 preparations 968:979 recombinant human immunoglobulin preparations 935:979 recombinant human immunoglobulin preparations 935:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 26 attach presence 899:906 arg2 species 924:930 high-mannose species 911:930 high-mannose species 911:930 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 7 27 theme human 1735:1739 arg1 antibodies 1741:1750 multiple recombinant human antibodies 1714:1750 multiple recombinant human antibodies 1714:1750 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 4 28 theme hamster 1084:1090 arg1 ovary 1092:1096 Chinese hamster ovary 1076:1096 Chinese hamster ovary cell culture 1076:1109 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 2 29 link O-linked 520:527 arg1 glycans 530:536 asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 462:536 glycans 530:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 6 30 theme dendritic 1501:1509 arg1 uptake 1516:1521 dendritic cell uptake 1501:1521 dendritic cell uptake 1501:1521 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 5 31 theme line 1320:1323 arg1 engineering 1325:1335 cell line engineering 1315:1335 cell line engineering 1315:1335 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 6 32 theme ADCC 1596:1599 arg1 levels 1586:1591 the levels 1582:1591 the levels of ADCC 1582:1599 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 1 33 theme paramount 118:126 arg1 modification 147:158 the paramount post-translational modification 114:158 the paramount post-translational modification on recombinant glycoproteins 114:187 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 34 theme used 1173:1176 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 34 theme used 1173:1176 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 5 35 with replacement 1222:1232 arg1 arabinose 1249:1257 arabinose 1249:1257 arabinose 1249:1257 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 0 36 theme recombinant 19:29 arg1 therapeutics 66:77 recombinant human immunoglobulin-based protein therapeutics 19:77 recombinant human immunoglobulin-based protein therapeutics 19:77 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 28375048 5 37 theme cell 1315:1318 arg1 engineering 1325:1335 cell line engineering 1315:1335 cell line engineering 1315:1335 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 4 38 theme species 924:930 arg1 presence 899:906 the presence 895:906 the presence of high-mannose species in recombinant human immunoglobulin preparations 895:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 0 39 theme immunoglobulin-based 37:56 arg1 therapeutics 66:77 recombinant human immunoglobulin-based protein therapeutics 19:77 recombinant human immunoglobulin-based protein therapeutics 19:77 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 28375048 7 40 theme functional 1689:1698 arg1 attributes 1700:1709 various structural or functional attributes 1667:1709 various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig 1667:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 3 41 contain have 593:596 arg1 levels 567:572 fucose levels 560:572 fucose levels 560:572 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 3 41 contain have 593:596 arg2 relationship 609:620 an inverse relationship 598:620 an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity 598:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 2 42 link asparagine-linked 462:478 arg1 N-linked 481:488 N-linked 481:488 N-linked 481:488 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 4 43 theme recombinant 935:945 arg1 preparations 968:979 recombinant human immunoglobulin preparations 935:979 recombinant human immunoglobulin preparations 935:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 44 theme fucose 560:565 arg1 levels 567:572 fucose levels 560:572 fucose levels 560:572 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 3 45 with relationship 609:620 arg1 degree 631:636 the degree 627:636 the degree of antibody-dependent cell-mediated cytotoxicity 627:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 7 46 theme structural 1675:1684 arg1 attributes 1700:1709 various structural or functional attributes 1667:1709 various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig 1667:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 1 47 from modification 147:158 arg1 glycoproteins 175:187 recombinant glycoproteins 163:187 recombinant glycoproteins 163:187 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 8 48 theme Protein 1777:1783 arg1 arabinosylation 1785:1799 Protein arabinosylation 1777:1799 Protein arabinosylation 1777:1799 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 2 49 theme diverse 445:451 arg1 range 453:457 a diverse range 443:457 a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 443:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 8 50 theme mammalian 1849:1857 arg1 glycoproteins 1869:1881 mammalian expressed glycoproteins 1849:1881 mammalian expressed glycoproteins 1849:1881 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 7 51 theme adverse 1648:1654 arg1 changes 1656:1662 any adverse changes 1644:1662 any adverse changes 1644:1662 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 4 52 theme cell 1188:1191 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 52 theme cell 1188:1191 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 53 theme favorable 799:807 arg1 profiles 825:832 less favorable pharmacokinetic profiles 794:832 less favorable pharmacokinetic profiles 794:832 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 1 54 theme glycoprotein 301:312 arg1 therapeutics 314:325 recombinant glycoprotein therapeutics 289:325 recombinant glycoprotein therapeutics 289:325 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 55 theme sugar 1201:1205 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 55 theme sugar 1201:1205 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 6 56 theme accompanying 1426:1437 arg1 reduction 1439:1447 the accompanying reduction 1422:1447 the accompanying reduction in high mannose glycans 1422:1471 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 3 57 theme inverse 601:607 arg1 relationship 609:620 an inverse relationship 598:620 an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity 598:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 5 58 theme cellular 1340:1347 arg1 adaptation 1349:1358 cellular adaptation 1340:1358 cellular adaptation 1340:1358 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 3 59 link N-linked 542:549 arg1 glycans 551:557 N-linked glycans 542:557 N-linked glycans 542:557 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 2 60 theme human 345:349 arg1 immunoglobulins 351:365 human immunoglobulins 345:365 human immunoglobulins 345:365 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 0 61 theme therapeutics 66:77 arg1 Arabinosylation 0:14 Arabinosylation 0:14 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 0:78 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 28375048 4 62 theme human 947:951 arg1 preparations 968:979 recombinant human immunoglobulin preparations 935:979 recombinant human immunoglobulin preparations 935:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 8 63 gly glycoproteins 1869:1881 arg1 glycoproteins 1869:1881 mammalian expressed glycoproteins 1849:1881 mammalian expressed glycoproteins 1849:1881 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 6 64 theme recombinant 1401:1411 arg1 IgGs 1413:1416 Arabinosylated recombinant IgGs 1386:1416 Arabinosylated recombinant IgGs 1386:1416 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 2 65 theme metabolic 404:412 arg1 pathway 414:420 a reasonably well-understood metabolic pathway 375:420 a reasonably well-understood metabolic pathway 375:420 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 4 66 with supplementation 1125:1139 arg1 D-arabinose 1146:1156 D-arabinose 1146:1156 D-arabinose 1146:1156 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 66 with supplementation 1125:1139 arg1 substrate 1207:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 an uncommonly used mammalian cell culture sugar substrate 1159:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 67 theme cytotoxicity 674:685 arg1 degree 631:636 the degree 627:636 the degree of antibody-dependent cell-mediated cytotoxicity 627:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 4 68 theme fucosylation 1039:1050 arg1 replacement 1024:1034 an approximate 100% replacement 1004:1034 an approximate 100% replacement of fucosylation with arabinosylation 1004:1071 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 3 69 theme antibody-dependent 641:658 arg1 cytotoxicity 674:685 antibody-dependent cell-mediated cytotoxicity 641:685 antibody-dependent cell-mediated cytotoxicity 641:685 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 6 70 from IgGs 1413:1416 arg1 glycans 1465:1471 high mannose glycans 1452:1471 high mannose glycans 1452:1471 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 6 71 theme mannose 1457:1463 arg1 glycans 1465:1471 high mannose glycans 1452:1471 high mannose glycans 1452:1471 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 3 72 theme mannose 697:703 arg1 levels 705:710 high mannose levels 692:710 high mannose levels 692:710 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 7 73 theme DVD-Ig 1769:1774 arg1 attributes 1700:1709 various structural or functional attributes 1667:1709 various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig 1667:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 4 74 theme 100 1019:1021 arg1 % 1022:1022 % 1022:1022 % 1022:1022 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 1 75 gly glycosylation 88:100 arg1 literature 214:223 the literature 210:223 the literature 210:223 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 76 from presence 899:906 arg1 preparations 968:979 recombinant human immunoglobulin preparations 935:979 recombinant human immunoglobulin preparations 935:979 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 77 theme ovary 1092:1096 arg1 culture 1103:1109 Chinese hamster ovary cell culture 1076:1109 Chinese hamster ovary cell culture 1076:1109 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 6 78 theme FcγRIIIa 1534:1541 arg1 signaling 1543:1551 FcγRIIIa signaling 1534:1551 FcγRIIIa signaling 1534:1551 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 7 79 theme antibodies 1741:1750 arg1 attributes 1700:1709 various structural or functional attributes 1667:1709 various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig 1667:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 1 80 theme Protein 80:86 arg1 glycosylation 88:100 Protein glycosylation 80:100 Protein glycosylation 80:100 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 7 81 theme recombinant 1723:1733 arg1 antibodies 1741:1750 multiple recombinant human antibodies 1714:1750 multiple recombinant human antibodies 1714:1750 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 4 82 theme novel 858:862 arg1 approach 864:871 a novel approach 856:871 a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate 856:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 6 83 theme cell 1511:1514 arg1 uptake 1516:1521 dendritic cell uptake 1501:1521 dendritic cell uptake 1501:1521 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 5 84 theme fucose 1237:1242 arg1 effective 1268:1276 effective 1268:1276 effective 1268:1276 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 5 84 theme fucose 1237:1242 arg1 replacement 1222:1232 The replacement 1218:1232 The replacement of fucose with arabinose 1218:1257 The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. 28375048 4 85 theme Chinese 1076:1082 arg1 ovary 1092:1096 Chinese hamster ovary 1076:1096 Chinese hamster ovary cell culture 1076:1109 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 1 86 gly glycoprotein 301:312 arg1 glycoprotein 301:312 recombinant glycoprotein therapeutics 289:325 recombinant glycoprotein therapeutics 289:325 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 4 87 with replacement 1024:1034 arg1 arabinosylation 1057:1071 arabinosylation 1057:1071 arabinosylation 1057:1071 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 2 88 theme O-linked 520:527 arg1 glycans 530:536 asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 462:536 glycans 530:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 0 89 theme human 31:35 arg1 therapeutics 66:77 recombinant human immunoglobulin-based protein therapeutics 19:77 recombinant human immunoglobulin-based protein therapeutics 19:77 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 28375048 1 90 theme post-translational 128:145 arg1 modification 147:158 the paramount post-translational modification 114:158 the paramount post-translational modification on recombinant glycoproteins 114:187 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 2 91 theme serine/threonine-linked 495:517 arg1 glycans 530:536 asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 462:536 glycans 530:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 3 92 theme N-linked 542:549 arg1 glycans 551:557 N-linked glycans 542:557 N-linked glycans 542:557 In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. 28375048 0 93 theme protein 58:64 arg1 therapeutics 66:77 recombinant human immunoglobulin-based protein therapeutics 19:77 recombinant human immunoglobulin-based protein therapeutics 19:77 Arabinosylation of recombinant human immunoglobulin-based protein therapeutics. 28375048 6 94 from reduction 1488:1496 arg1 uptake 1516:1521 dendritic cell uptake 1501:1521 dendritic cell uptake 1501:1521 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 28375048 7 95 theme multiple 1714:1721 arg1 antibodies 1741:1750 multiple recombinant human antibodies 1714:1750 multiple recombinant human antibodies 1714:1750 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 4 96 theme media 1119:1123 arg1 supplementation 1125:1139 media supplementation 1119:1139 media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate 1119:1215 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 4 97 theme high-mannose 911:922 arg1 species 924:930 high-mannose species 911:930 high-mannose species 911:930 Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. 28375048 2 98 theme N-linked 481:488 arg1 range 453:457 a diverse range 443:457 a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 443:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 2 99 theme glycans 530:536 arg1 range 453:457 a diverse range 443:457 a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans 443:536 Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. 28375048 8 100 theme expressed 1859:1867 arg1 glycoproteins 1869:1881 mammalian expressed glycoproteins 1849:1881 mammalian expressed glycoproteins 1849:1881 Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins. 28375048 7 101 theme various 1667:1673 arg1 attributes 1700:1709 various structural or functional attributes 1667:1709 various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig 1667:1774 These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. 28375048 1 102 theme recombinant 289:299 arg1 therapeutics 314:325 recombinant glycoprotein therapeutics 289:325 recombinant glycoprotein therapeutics 289:325 Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. 28375048 6 103 from reduction 1439:1447 arg1 glycans 1465:1471 high mannose glycans 1452:1471 high mannose glycans 1452:1471 Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. 25378480 9 0 theme minimal 1833:1839 arg1 changes 1841:1847 minimal changes 1833:1847 minimal changes between phylogenetically distant protein structures 1833:1899 More generally, this work demonstrates that it is possible to exchange the specificities and catalytic mechanisms with minimal changes between phylogenetically distant protein structures. 25378480 0 1 from design 8:13 arg1 β-glycosidase 72:84 a GH1 β-glycosidase 66:84 a GH1 β-glycosidase by mechanism engineering 66:109 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 7 2 from 84 1472:1473 arg1 N-acetylglucosaminidases 1426:1449 the N-acetylglucosaminidases 1422:1449 the N-acetylglucosaminidases from Families 20 and 84 1422:1473 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 1 3 theme glycoconjugates 257:271 arg1 degradation 214:224 the degradation 210:224 the degradation of natural polysaccharides and glycoconjugates 210:271 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 5 4 with conflicts 997:1005 arg1 E338 1043:1046 E338 1043:1046 E338 1043:1046 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 4 with conflicts 997:1005 arg1 site 1019:1022 active site amino-acids 1012:1034 active site amino-acids (N163, E338) induced by the N-acetyl group 1012:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 5 theme N-acetyl 1064:1071 arg1 group 1073:1077 the N-acetyl group 1060:1077 the N-acetyl group 1060:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 1 6 from areas 166:170 arg1 abundant 150:157 abundant 150:157 abundant 150:157 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 3 7 theme enzymes 514:520 arg1 relationships 491:503 the structure/function relationships 468:503 the structure/function relationships of these enzymes and their evolution 468:540 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 8 8 theme double 1567:1572 arg1 donor 1633:1637 donor 1633:1637 donor 1633:1637 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 8 8 theme double 1567:1572 arg1 mutant 1574:1579 the double mutant 1563:1579 the double mutant 1563:1579 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 8 8 theme double 1567:1572 arg1 N-methyl-O-benzyl-N- 1643:1662 N-methyl-O-benzyl-N- 1643:1662 N-methyl-O-benzyl-N- 1643:1662 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 5 9 from subsite 905:911 arg1 thermophilus 950:961 Thermus thermophilus 942:961 Thermus thermophilus (TtβGly) 942:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 9 from subsite 905:911 arg1 TtβGly 964:969 TtβGly 964:969 TtβGly 964:969 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 4 10 theme CAZy 785:788 arg1 Families 790:797 CAZy Families 3, 20 and 84 785:810 CAZy Families 3, 20 and 84 785:810 This latter activity is totally absent in Family 1, while natural β-hexosaminidases belong to CAZy Families 3, 20 and 84. 25378480 3 11 theme robust 561:566 arg1 glycosidases 576:587 new robust evolved glycosidases 557:587 new robust evolved glycosidases 557:587 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 0 12 theme mechanism 89:97 arg1 engineering 99:109 mechanism engineering 89:109 mechanism engineering 89:109 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 5 13 theme β-glycosidase 923:935 arg1 subsite 905:911 the subsite -1 901:914 the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) 901:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 13 theme β-glycosidase 923:935 arg1 β-glycosidase 923:935 the β-glycosidase 919:935 the β-glycosidase from Thermus thermophilus (TtβGly) 919:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 9 14 theme distant 1874:1880 arg1 structures 1890:1899 phylogenetically distant protein structures 1857:1899 phylogenetically distant protein structures 1857:1899 More generally, this work demonstrates that it is possible to exchange the specificities and catalytic mechanisms with minimal changes between phylogenetically distant protein structures. 25378480 8 15 dep donor 1633:1637 arg1 a 1631:1631 a 1631:1631 a 1631:1631 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 4 16 theme natural 749:755 arg1 β-hexosaminidases 757:773 natural β-hexosaminidases 749:773 natural β-hexosaminidases 749:773 This latter activity is totally absent in Family 1, while natural β-hexosaminidases belong to CAZy Families 3, 20 and 84. 25378480 3 17 theme evolution 532:540 arg1 relationships 491:503 the structure/function relationships 468:503 the structure/function relationships of these enzymes and their evolution 468:540 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 7 18 from Families 1456:1463 arg1 N-acetylglucosaminidases 1426:1449 the N-acetylglucosaminidases 1422:1449 the N-acetylglucosaminidases from Families 20 and 84 1422:1473 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 1 19 theme metabolism 175:184 arg1 areas 166:170 all areas 162:170 all areas of metabolism 162:184 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 9 20 theme protein 1882:1888 arg1 structures 1890:1899 phylogenetically distant protein structures 1857:1899 phylogenetically distant protein structures 1857:1899 More generally, this work demonstrates that it is possible to exchange the specificities and catalytic mechanisms with minimal changes between phylogenetically distant protein structures. 25378480 7 21 theme mutant 1192:1197 arg1 N163D-E338G 1199:1209 The double mutant N163D-E338G 1181:1209 The double mutant N163D-E338G 1181:1209 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 7 21 theme mutant 1192:1197 arg1 active 1220:1225 active 1220:1225 active 1220:1225 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 5 22 theme several 982:988 arg1 conflicts 997:1005 several steric conflicts 982:1005 several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group 982:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 8 23 theme trans-N-acetylglucosaminidase 1506:1534 arg1 activity 1536:1543 a very efficient trans-N-acetylglucosaminidase activity 1489:1543 a very efficient trans-N-acetylglucosaminidase activity 1489:1543 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 2 24 theme family 381:386 arg1 members 365:371 members 365:371 members of each family 365:386 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 7 25 theme double 1185:1190 arg1 N163D-E338G 1199:1209 The double mutant N163D-E338G 1181:1209 The double mutant N163D-E338G 1181:1209 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 7 25 theme double 1185:1190 arg1 active 1220:1225 active 1220:1225 active 1220:1225 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 1 26 from abundant 150:157 arg1 areas 166:170 all areas 162:170 all areas of metabolism 162:184 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 3 27 theme evolved 568:574 arg1 glycosidases 576:587 new robust evolved glycosidases 557:587 new robust evolved glycosidases 557:587 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 7 28 theme mutated 1285:1291 arg1 enzyme 1293:1298 this mutated enzyme 1280:1298 this mutated enzyme 1280:1298 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 4 29 theme latter 696:701 arg1 activity 703:710 This latter activity 691:710 This latter activity 691:710 This latter activity is totally absent in Family 1, while natural β-hexosaminidases belong to CAZy Families 3, 20 and 84. 25378480 0 30 theme De 0:1 arg1 design 8:13 De novo design 0:13 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 0:110 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 2 31 theme CAZy 322:325 arg1 http 335:338 http 335:338 http 335:338 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 2 31 theme CAZy 322:325 arg1 server 327:332 CAZy server 322:332 CAZy server 322:332 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 7 32 theme substrate-assisted 1339:1356 arg1 catalysis 1358:1366 a substrate-assisted catalysis 1337:1366 a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84 1337:1473 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 5 33 theme molecular 819:827 arg1 modeling 829:836 molecular modeling 819:836 molecular modeling 819:836 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 7 34 theme noncovalent 1375:1385 arg1 intermediate 1397:1408 a noncovalent oxazoline intermediate 1373:1408 a noncovalent oxazoline intermediate 1373:1408 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 3 35 theme new 557:559 arg1 glycosidases 576:587 new robust evolved glycosidases 557:587 new robust evolved glycosidases 557:587 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 2 36 theme catalytic 417:425 arg1 mechanism 427:435 catalytic mechanism 417:435 catalytic mechanism 417:435 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 8 37 theme efficient 1496:1504 arg1 trans-N-acetylglucosaminidase 1506:1534 a very efficient trans-N-acetylglucosaminidase 1489:1534 a very efficient trans-N-acetylglucosaminidase activity 1489:1543 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 1 38 theme Glycoside 112:120 arg1 hydrolases 122:131 Glycoside hydrolases 112:131 Glycoside hydrolases 112:131 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 9 39 with exchange 1776:1783 arg1 changes 1841:1847 minimal changes 1833:1847 minimal changes between phylogenetically distant protein structures 1833:1899 More generally, this work demonstrates that it is possible to exchange the specificities and catalytic mechanisms with minimal changes between phylogenetically distant protein structures. 25378480 0 40 theme trans-β-N-acetylglucosaminidase 20:50 arg1 activity 52:59 a trans-β-N-acetylglucosaminidase activity 18:59 a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering 18:109 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 8 41 theme NAG-oxazoline 1614:1626 arg1 presence 1602:1609 the presence 1598:1609 the presence of NAG-oxazoline 1598:1626 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 0 42 from β-glycosidase 72:84 arg1 design 8:13 De novo design 0:13 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 0:110 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 0 42 from β-glycosidase 72:84 arg1 activity 52:59 a trans-β-N-acetylglucosaminidase activity 18:59 a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering 18:109 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 2 43 theme similar 395:401 arg1 structure 403:411 a similar structure 393:411 a similar structure 393:411 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 3 44 theme thermostable 625:636 arg1 β-glycosidase 638:650 a Family 1 thermostable β-glycosidase 614:650 a Family 1 thermostable β-glycosidase 614:650 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 5 45 theme amino-acids 1024:1034 arg1 E338 1043:1046 E338 1043:1046 E338 1043:1046 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 45 theme amino-acids 1024:1034 arg1 site 1019:1022 active site amino-acids 1012:1034 active site amino-acids (N163, E338) induced by the N-acetyl group 1012:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 7 46 theme catalytic 1307:1315 arg1 mechanism 1317:1325 a catalytic mechanism 1305:1325 a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84 1305:1473 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 2 47 contain have 388:391 arg2 structure 403:411 a similar structure 393:411 a similar structure 393:411 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 2 47 contain have 388:391 arg2 mechanism 427:435 catalytic mechanism 417:435 catalytic mechanism 417:435 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 2 47 contain have 388:391 arg1 members 365:371 members 365:371 members of each family 365:386 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 6 48 theme significant 1125:1135 arg1 activity 1161:1168 significant N-acetylglucosaminidase activity 1125:1168 significant N-acetylglucosaminidase activity 1125:1168 Both N163A and N163D-E338G mutations induced significant N-acetylglucosaminidase activity in TtβGly. 25378480 0 49 theme activity 52:59 arg1 design 8:13 De novo design 0:13 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 0:110 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 7 50 theme bicyclic 1234:1241 arg1 substrate 1253:1261 the bicyclic oxazoline substrate 1230:1261 the bicyclic oxazoline substrate 1230:1261 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 7 51 theme similar 1411:1417 arg1 intermediate 1397:1408 a noncovalent oxazoline intermediate 1373:1408 a noncovalent oxazoline intermediate 1373:1408 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 2 52 dep families 312:319 arg1 http 335:338 http 335:338 http 335:338 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 2 52 dep families 312:319 arg1 server 327:332 CAZy server 322:332 CAZy server 322:332 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 5 53 from thermophilus 950:961 arg1 subsite 905:911 the subsite -1 901:914 the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) 901:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 53 from thermophilus 950:961 arg1 β-glycosidase 923:935 the β-glycosidase 919:935 the β-glycosidase from Thermus thermophilus (TtβGly) 919:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 54 theme active 1012:1017 arg1 E338 1043:1046 E338 1043:1046 E338 1043:1046 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 54 theme active 1012:1017 arg1 site 1019:1022 active site amino-acids 1012:1034 active site amino-acids (N163, E338) induced by the N-acetyl group 1012:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 7 55 with catalysis 1358:1366 arg1 intermediate 1397:1408 a noncovalent oxazoline intermediate 1373:1408 a noncovalent oxazoline intermediate 1373:1408 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 5 56 theme steric 990:995 arg1 conflicts 997:1005 several steric conflicts 982:1005 several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group 982:1077 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 3 57 theme Family 616:621 arg1 β-glycosidase 638:650 a Family 1 thermostable β-glycosidase 614:650 a Family 1 thermostable β-glycosidase 614:650 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 2 58 dep server 327:332 arg1 //www.cazy.org 340:353 //www.cazy.org 340:353 //www.cazy.org 340:353 These enzymes are classified into 133 families (CAZy server, http://www.cazy.org) in which members of each family have a similar structure and catalytic mechanism. 25378480 6 59 theme N163D-E338G 1095:1105 arg1 mutations 1107:1115 N163D-E338G mutations 1095:1115 N163D-E338G mutations 1095:1115 Both N163A and N163D-E338G mutations induced significant N-acetylglucosaminidase activity in TtβGly. 25378480 9 60 theme catalytic 1807:1815 arg1 mechanisms 1817:1826 catalytic mechanisms 1807:1826 catalytic mechanisms 1807:1826 More generally, this work demonstrates that it is possible to exchange the specificities and catalytic mechanisms with minimal changes between phylogenetically distant protein structures. 25378480 7 61 theme oxazoline 1387:1395 arg1 intermediate 1397:1408 a noncovalent oxazoline intermediate 1373:1408 a noncovalent oxazoline intermediate 1373:1408 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 5 62 from docking 864:870 arg1 subsite 905:911 the subsite -1 901:914 the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) 901:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 5 62 from docking 864:870 arg1 β-glycosidase 923:935 the β-glycosidase 919:935 the β-glycosidase from Thermus thermophilus (TtβGly) 919:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 3 63 theme structure/function 472:489 arg1 relationships 491:503 the structure/function relationships 468:503 the structure/function relationships of these enzymes and their evolution 468:540 In order to understand better the structure/function relationships of these enzymes and their evolution and to develop new robust evolved glycosidases, we undertook to convert a Family 1 thermostable β-glycosidase into an exo-β-N-acetylglucosaminidase. 25378480 4 64 from Family 733:738 arg1 absent 723:728 absent 723:728 absent 723:728 This latter activity is totally absent in Family 1, while natural β-hexosaminidases belong to CAZy Families 3, 20 and 84. 25378480 1 65 theme natural 229:235 arg1 polysaccharides 237:251 natural polysaccharides 229:251 natural polysaccharides 229:251 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 0 66 theme GH1 68:70 arg1 β-glycosidase 72:84 a GH1 β-glycosidase 66:84 a GH1 β-glycosidase by mechanism engineering 66:109 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 8 67 theme -hydroxylamine 1683:1696 arg1 donor 1633:1637 donor 1633:1637 donor 1633:1637 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 8 67 theme -hydroxylamine 1683:1696 arg1 mutant 1574:1579 the double mutant 1563:1579 the double mutant 1563:1579 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 8 67 theme -hydroxylamine 1683:1696 arg1 β-d-glucopyranosyl 1664:1681 β-d-glucopyranosyl 1664:1681 β-d-glucopyranosyl 1664:1681 Furthermore, a very efficient trans-N-acetylglucosaminidase activity was observed when the double mutant was incubated in the presence of NAG-oxazoline as a donor and N-methyl-O-benzyl-N-(β-d-glucopyranosyl)-hydroxylamine as an acceptor. 25378480 5 68 theme N-acetyl-d-glucosamine 875:896 arg1 docking 864:870 the docking 860:870 the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) 860:970 Using molecular modeling, we first showed that the docking of N-acetyl-d-glucosamine in the subsite -1 of the β-glycosidase from Thermus thermophilus (TtβGly) suggested several steric conflicts with active site amino-acids (N163, E338) induced by the N-acetyl group. 25378480 1 69 theme polysaccharides 237:251 arg1 degradation 214:224 the degradation 210:224 the degradation of natural polysaccharides and glycoconjugates 210:271 Glycoside hydrolases are particularly abundant in all areas of metabolism as they are involved in the degradation of natural polysaccharides and glycoconjugates. 25378480 7 70 theme oxazoline 1243:1251 arg1 substrate 1253:1261 the bicyclic oxazoline substrate 1230:1261 the bicyclic oxazoline substrate 1230:1261 The double mutant N163D-E338G was also active on the bicyclic oxazoline substrate, suggesting that this mutated enzyme uses a catalytic mechanism involving a substrate-assisted catalysis with a noncovalent oxazoline intermediate, similar to the N-acetylglucosaminidases from Families 20 and 84. 25378480 6 71 theme N-acetylglucosaminidase 1137:1159 arg1 activity 1161:1168 significant N-acetylglucosaminidase activity 1125:1168 significant N-acetylglucosaminidase activity 1125:1168 Both N163A and N163D-E338G mutations induced significant N-acetylglucosaminidase activity in TtβGly. 25378480 0 72 dep De 0:1 arg1 novo 3:6 novo 3:6 novo 3:6 De novo design of a trans-β-N-acetylglucosaminidase activity from a GH1 β-glycosidase by mechanism engineering. 25378480 4 73 from absent 723:728 arg1 Family 733:738 Family 1 733:740 Family 1 733:740 This latter activity is totally absent in Family 1, while natural β-hexosaminidases belong to CAZy Families 3, 20 and 84. 26138399 9 0 theme prebiotic 1698:1706 arg1 N-glycans 1708:1716 prebiotic N-glycans 1698:1716 prebiotic N-glycans 1698:1716 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 8 1 theme glycans 1528:1534 arg1 release 1498:1504 large-scale release 1486:1504 large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1486:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 5 2 theme model 694:698 arg1 lactoferrin 737:747 bovine lactoferrin 730:747 bovine lactoferrin 730:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 2 theme model 694:698 arg1 substrates 700:709 the model substrates ribonuclease B and bovine lactoferrin 690:747 the model substrates ribonuclease B and bovine lactoferrin 690:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 2 theme model 694:698 arg1 B 724:724 ribonuclease B 711:724 ribonuclease B 711:724 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 2 3 theme gut 390:392 arg1 subsp 426:430 Bifidobacterium longum subsp 403:430 Bifidobacterium longum subsp 403:430 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 2 3 theme gut 390:392 arg1 microbe 394:400 a key infant gut microbe 377:400 a key infant gut microbe 377:400 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 1 4 theme mannose 284:290 arg1 N-glycans 312:320 high mannose, hybrid and complex N-glycans 279:320 N-glycans 312:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 9 5 theme N-glycans 1708:1716 arg1 source 1688:1693 a source 1686:1693 a source of prebiotic N-glycans 1686:1716 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 9 5 theme N-glycans 1708:1716 arg1 whey 1616:1619 whey 1616:1619 whey 1616:1619 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 0 6 from characterization 8:23 arg1 whey 100:103 concentrated bovine colostrum whey 70:103 concentrated bovine colostrum whey to release bioactive glycans 70:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 2 7 theme infant 383:388 arg1 subsp 426:430 Bifidobacterium longum subsp 403:430 Bifidobacterium longum subsp 403:430 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 2 7 theme infant 383:388 arg1 microbe 394:400 a key infant gut microbe 377:400 a key infant gut microbe 377:400 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 0 8 theme bioactive 116:124 arg1 glycans 126:132 release bioactive glycans 108:132 release bioactive glycans 108:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 5 9 theme ribonuclease 711:722 arg1 substrates 700:709 the model substrates ribonuclease B and bovine lactoferrin 690:747 the model substrates ribonuclease B and bovine lactoferrin 690:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 9 theme ribonuclease 711:722 arg1 B 724:724 ribonuclease B 711:724 ribonuclease B 711:724 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 10 theme kinetic 655:661 arg1 parameters 663:672 kinetic parameters 655:672 kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey 655:797 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 1 11 theme hybrid 293:298 arg1 N-glycans 312:320 high mannose, hybrid and complex N-glycans 279:320 N-glycans 312:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 0 12 theme release 108:114 arg1 glycans 126:132 release bioactive glycans 108:132 release bioactive glycans 108:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 8 13 theme large-scale 1486:1496 arg1 release 1498:1504 large-scale release 1486:1504 large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1486:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 4 14 theme broad 447:451 arg1 specificity 453:463 The broad specificity 443:463 The broad specificity of EndoBI-1 443:475 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 5 15 theme colostrum 784:792 arg1 whey 794:797 concentrated bovine colostrum whey 764:797 concentrated bovine colostrum whey 764:797 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 8 16 theme phenol-sulphuric 1263:1278 arg1 assay 1299:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 2 17 theme longum 419:424 arg1 subsp 426:430 Bifidobacterium longum subsp 403:430 Bifidobacterium longum subsp 403:430 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 2 17 theme longum 419:424 arg1 microbe 394:400 a key infant gut microbe 377:400 a key infant gut microbe 377:400 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 9 18 used used 1666:1669 arg2 source 1688:1693 a source 1686:1693 a source of prebiotic N-glycans 1686:1716 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 9 18 used used 1666:1669 arg2 whey 1616:1619 whey 1616:1619 whey 1616:1619 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 0 19 from endo-β-N-acetylglucosaminidase 36:65 arg1 whey 100:103 concentrated bovine colostrum whey 70:103 concentrated bovine colostrum whey to release bioactive glycans 70:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 8 20 from variety 1543:1549 arg1 release 1498:1504 large-scale release 1486:1504 large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1486:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 7 21 theme highest 1097:1103 arg1 affinity 1114:1121 the highest apparent affinity 1093:1121 the highest apparent affinity for ribonuclease B 1093:1140 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 6 22 dep 0.49 831:834 arg1 to 828:829 to 828:829 to 828:829 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 4 23 theme many 515:518 arg1 applications 520:531 many applications 515:531 many applications 515:531 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 4 24 theme milk 567:570 arg1 glycoproteins 572:584 milk glycoproteins 567:584 milk glycoproteins in dairy processing 567:604 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 5 25 theme commercial 625:634 arg1 use 636:638 its commercial use 621:638 its commercial use 621:638 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 2 26 theme prebiotic 354:362 arg1 activity 364:371 selective prebiotic activity 344:371 selective prebiotic activity 344:371 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 1 27 theme N-N'-diacetyl 217:229 arg1 moiety 242:247 the N-N'-diacetyl chitobiose moiety 213:247 the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans 213:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 0 28 theme Kinetic 0:6 arg1 characterization 8:23 Kinetic characterization 0:23 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 0:133 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 1 29 theme complex 304:310 arg1 N-glycans 312:320 high mannose, hybrid and complex N-glycans 279:320 N-glycans 312:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 6 30 dep -3 936:937 arg1 to 940:941 to 940:941 to 940:941 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 30 dep -3 936:937 arg1 -3 951:952 -3 951:952 -3 951:952 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 30 dep -3 936:937 arg1 1.9×10 959:964 1.9×10(-2) 959:968 1.9×10(-2) 959:968 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 30 dep -3 936:937 arg1 7.75×10 943:949 7.75×10 943:949 7.75×10 943:949 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 8 31 dep suggest 1454:1460 arg1 facilitate 1475:1484 facilitate 1475:1484 suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1454:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 2 32 theme selective 344:352 arg1 activity 364:371 selective prebiotic activity 344:371 selective prebiotic activity 344:371 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 5 33 theme concentrated 764:775 arg1 whey 794:797 concentrated bovine colostrum whey 764:797 concentrated bovine colostrum whey 764:797 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 1 34 theme chitobiose 231:240 arg1 moiety 242:247 the N-N'-diacetyl chitobiose moiety 213:247 the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans 213:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 7 35 theme apparent 1105:1112 arg1 affinity 1114:1121 the highest apparent affinity 1093:1121 the highest apparent affinity for ribonuclease B 1093:1140 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 1 36 theme N-glycans 312:320 arg1 core 271:274 the N-glycan core 258:274 the N-glycan core of high mannose, hybrid and complex N-glycans 258:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 8 37 dep complex 1509:1515 arg1 bioactive 1518:1526 bioactive 1518:1526 bioactive 1518:1526 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 4 38 from glycoproteins 572:584 arg1 processing 595:604 dairy processing 589:604 dairy processing 589:604 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 8 39 theme resultant 1313:1321 arg1 structures 1332:1341 the resultant N-glycan structures 1309:1341 the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1309:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 6 40 theme 5.2×10 972:977 arg1 mg/mL×min 983:991 5.2×10(-2) mg/mL×min 972:991 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B 929:1026 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 5 41 dep substrates 700:709 arg1 lactoferrin 737:747 bovine lactoferrin 730:747 bovine lactoferrin 730:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 41 dep substrates 700:709 arg1 substrates 700:709 the model substrates ribonuclease B and bovine lactoferrin 690:747 the model substrates ribonuclease B and bovine lactoferrin 690:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 41 dep substrates 700:709 arg1 B 724:724 ribonuclease B 711:724 ribonuclease B 711:724 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 6 42 dep 3.18 862:865 arg1 to 859:860 to 859:860 to 859:860 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 43 theme Vmax 877:880 arg1 values 882:887 Vmax values 877:887 Vmax values 877:887 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 8 44 theme kinetic 1383:1389 arg1 parameters 1391:1400 The kinetic parameters 1379:1400 The kinetic parameters 1379:1400 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 6 45 dep 1.00 845:848 arg1 to 842:843 to 842:843 to 842:843 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 46 theme ribonuclease 1013:1024 arg1 B 1026:1026 ribonuclease B 1013:1026 ribonuclease B 1013:1026 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 0 47 theme endo-β-N-acetylglucosaminidase 36:65 arg1 characterization 8:23 Kinetic characterization 0:23 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 0:133 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 9 48 theme waste 1644:1648 arg1 product 1650:1656 a waste product 1642:1656 a waste product 1642:1656 Moreover, these results suggest that whey, often considered as a waste product, can be used effectively as a source of prebiotic N-glycans. 26138399 0 49 from whey 100:103 arg1 characterization 8:23 Kinetic characterization 0:23 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 0:133 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 8 50 gly glycoprotein 1554:1565 arg1 glycoprotein 1554:1565 glycoprotein substrates 1554:1576 glycoprotein substrates 1554:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 0 51 theme novel 30:34 arg1 endo-β-N-acetylglucosaminidase 36:65 a novel endo-β-N-acetylglucosaminidase 28:65 a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans 28:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 5 52 theme bovine 777:782 arg1 whey 794:797 concentrated bovine colostrum whey 764:797 concentrated bovine colostrum whey 764:797 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 0 53 theme concentrated 70:81 arg1 whey 100:103 concentrated bovine colostrum whey 70:103 concentrated bovine colostrum whey to release bioactive glycans 70:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 2 54 contain have 339:342 arg2 activity 364:371 selective prebiotic activity 344:371 selective prebiotic activity 344:371 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 2 54 contain have 339:342 arg1 N-glycans 329:337 These N-glycans 323:337 These N-glycans 323:337 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 8 55 theme carbohydrate 1286:1297 arg1 assay 1299:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 56 theme complex 1509:1515 arg1 glycans 1528:1534 complex, bioactive glycans 1509:1534 complex, bioactive glycans 1509:1534 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 57 theme total 1280:1284 arg1 assay 1299:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 a phenol-sulphuric total carbohydrate assay 1261:1303 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 2 58 theme key 379:381 arg1 subsp 426:430 Bifidobacterium longum subsp 403:430 Bifidobacterium longum subsp 403:430 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 2 58 theme key 379:381 arg1 microbe 394:400 a key infant gut microbe 377:400 a key infant gut microbe 377:400 These N-glycans have selective prebiotic activity for a key infant gut microbe, Bifidobacterium longum subsp. 26138399 4 59 theme dairy 589:593 arg1 processing 595:604 dairy processing 589:604 dairy processing 589:604 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 5 60 theme bovine 730:735 arg1 lactoferrin 737:747 bovine lactoferrin 730:747 bovine lactoferrin 730:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 60 theme bovine 730:735 arg1 substrates 700:709 the model substrates ribonuclease B and bovine lactoferrin 690:747 the model substrates ribonuclease B and bovine lactoferrin 690:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 1 61 located found 249:253 arg1 core 271:274 the N-glycan core 258:274 the N-glycan core of high mannose, hybrid and complex N-glycans 258:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 1 61 located found 249:253 arg2 moiety 242:247 the N-N'-diacetyl chitobiose moiety 213:247 the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans 213:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 8 62 theme N-glycan 1323:1330 arg1 structures 1332:1341 the resultant N-glycan structures 1309:1341 the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates 1309:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 6 63 theme Km 800:801 arg1 values 803:808 Km values 800:808 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL 800:871 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 7 64 theme ribonuclease 1127:1138 arg1 B 1140:1140 ribonuclease B 1127:1140 ribonuclease B 1127:1140 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 8 65 theme structural 1406:1415 arg1 characterization 1417:1432 structural characterization 1406:1432 structural characterization 1406:1432 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 66 theme substrates 1567:1576 arg1 variety 1543:1549 a variety 1541:1549 a variety of glycoprotein substrates 1541:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 66 theme substrates 1567:1576 arg1 substrates 1567:1576 glycoprotein substrates 1554:1576 glycoprotein substrates 1554:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 7 67 theme reaction 1161:1168 arg1 rate 1170:1173 the maximum reaction rate 1149:1173 the maximum reaction rate 1149:1173 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 7 67 theme reaction 1161:1168 arg1 highest 1183:1189 highest 1183:1189 highest 1183:1189 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 1 68 theme N-glycan 262:269 arg1 core 271:274 the N-glycan core 258:274 the N-glycan core of high mannose, hybrid and complex N-glycans 258:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 8 69 theme glycans 1437:1443 arg1 parameters 1391:1400 The kinetic parameters 1379:1400 The kinetic parameters 1379:1400 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 69 theme glycans 1437:1443 arg1 characterization 1417:1432 structural characterization 1406:1432 structural characterization 1406:1432 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 8 70 theme EndoBI-1-released 1214:1230 arg1 N-glycans 1232:1240 EndoBI-1-released N-glycans 1214:1240 EndoBI-1-released N-glycans 1214:1240 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 7 71 theme maximum 1153:1159 arg1 rate 1170:1173 the maximum reaction rate 1149:1173 the maximum reaction rate 1149:1173 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 7 71 theme maximum 1153:1159 arg1 highest 1183:1189 highest 1183:1189 highest 1183:1189 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 8 72 theme glycoprotein 1554:1565 arg1 substrates 1567:1576 glycoprotein substrates 1554:1576 glycoprotein substrates 1554:1576 EndoBI-1-released N-glycans were quantified by a phenol-sulphuric total carbohydrate assay and the resultant N-glycan structures monitored by nano-LC-Chip-Q-TOF MS. The kinetic parameters and structural characterization of glycans released suggest EndoBI-1 can facilitate large-scale release of complex, bioactive glycans from a variety of glycoprotein substrates. 26138399 0 73 theme colostrum 90:98 arg1 whey 100:103 concentrated bovine colostrum whey 70:103 concentrated bovine colostrum whey to release bioactive glycans 70:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 4 74 theme EndoBI-1 468:475 arg1 specificity 453:463 The broad specificity 443:463 The broad specificity of EndoBI-1 443:475 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 4 75 gly glycoproteins 572:584 arg1 glycoproteins 572:584 milk glycoproteins 567:584 milk glycoproteins in dairy processing 567:604 The broad specificity of EndoBI-1 suggests the enzyme may be useful for many applications, particularly for deglycosylating milk glycoproteins in dairy processing. 26138399 1 76 theme isolated 158:165 arg1 endo-β-N-acetylglucosaminidase 167:196 a recently isolated endo-β-N-acetylglucosaminidase 147:196 a recently isolated endo-β-N-acetylglucosaminidase 147:196 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 1 76 theme isolated 158:165 arg1 EndoBI-1 135:142 EndoBI-1 135:142 EndoBI-1 135:142 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26138399 7 77 theme concentrated 1195:1206 arg1 whey 1208:1211 concentrated whey 1195:1211 concentrated whey 1195:1211 In general, EndoBI-1 showed the highest apparent affinity for ribonuclease B, while the maximum reaction rate was the highest for concentrated whey. 26138399 0 78 theme bovine 83:88 arg1 whey 100:103 concentrated bovine colostrum whey 70:103 concentrated bovine colostrum whey to release bioactive glycans 70:132 Kinetic characterization of a novel endo-β-N-acetylglucosaminidase on concentrated bovine colostrum whey to release bioactive glycans. 26138399 6 79 dep 7.75×10 943:949 arg1 to 969:970 to 969:970 to 969:970 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 79 dep 7.75×10 943:949 arg1 mg/mL×min 983:991 5.2×10(-2) mg/mL×min 972:991 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B 929:1026 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 to 913:914 to 913:914 to 913:914 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 5.09×10 916:922 5.09×10 916:922 5.09×10 916:922 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 -3 936:937 -3 936:937 -3 936:937 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 whey 1045:1048 whey 1045:1048 whey 1045:1048 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 -3 924:925 -3 924:925 -3 924:925 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 6 80 dep 3.5×10 902:907 arg1 lactoferrin 1029:1039 lactoferrin 1029:1039 lactoferrin 1029:1039 Km values ranging from 0.25 to 0.49, 0.43 to 1.00 and 0.90 to 3.18 mg/mL and Vmax values ranging from 3.5×10(-3) to 5.09×10(-3), 4.5×10(-3) to 7.75×10(-3) and 1.9×10(-2)to 5.2×10(-2) mg/mL×min were determined for ribonuclease B, lactoferrin and whey, respectively. 26138399 5 81 from parameters 663:672 arg1 lactoferrin 737:747 bovine lactoferrin 730:747 bovine lactoferrin 730:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 81 from parameters 663:672 arg1 substrates 700:709 the model substrates ribonuclease B and bovine lactoferrin 690:747 the model substrates ribonuclease B and bovine lactoferrin 690:747 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 81 from parameters 663:672 arg1 whey 794:797 concentrated bovine colostrum whey 764:797 concentrated bovine colostrum whey 764:797 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 5 81 from parameters 663:672 arg1 B 724:724 ribonuclease B 711:724 ribonuclease B 711:724 To facilitate its commercial use, we determined kinetic parameters for EndoBI-1 on the model substrates ribonuclease B and bovine lactoferrin, as well as on concentrated bovine colostrum whey. 26138399 1 82 theme high 279:282 arg1 N-glycans 312:320 high mannose, hybrid and complex N-glycans 279:320 N-glycans 312:320 EndoBI-1 is a recently isolated endo-β-N-acetylglucosaminidase, which cleaves the N-N'-diacetyl chitobiose moiety found in the N-glycan core of high mannose, hybrid and complex N-glycans. 26537503 7 0 theme flexible 1423:1430 arg1 loops 1467:1471 flexible complementarity determining region loops 1423:1471 flexible complementarity determining region loops that sample the bound conformations occasionally 1423:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 6 1 dep V1/V2 1179:1183 arg1 the 1175:1177 the 1175:1177 the 1175:1177 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 7 2 with shield 1411:1416 arg1 loops 1467:1471 flexible complementarity determining region loops 1423:1471 flexible complementarity determining region loops that sample the bound conformations occasionally 1423:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 2 3 theme some 347:350 arg1 epitopes 335:342 the epitopes 331:342 the epitopes of some of these antibodies 331:370 Structural studies reveal that the epitopes of some of these antibodies involve one or more crucial glycans, without which the binding is completely abolished. 26537503 4 4 theme glycan-only 888:898 arg1 systems 900:906 glycan-only systems 888:906 glycan-only systems 888:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 0 5 from Roles 0:4 arg1 interactions 20:31 interactions 20:31 interactions between gp120 and HIV 20:53 Roles of glycans in interactions between gp120 and HIV broadly neutralizing antibodies. 26537503 3 6 theme glycans 518:524 arg1 roles 509:513 the critical roles 496:513 the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates 496:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 5 7 theme simulation 913:922 arg1 results 924:930 The simulation results 909:930 The simulation results 909:930 The simulation results show that the complex structures are stabilized by the glycans, and the multivalent interactions between the antibody and gp120 promote cooperativities to further enhance the binding. 26537503 4 8 theme antibody-gp120 793:806 arg1 complex 808:814 antibody-gp120 complex 793:814 antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 793:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 6 9 theme free 1123:1126 arg1 gp120 1128:1132 the free gp120 1119:1132 the free gp120 1119:1132 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 6 10 theme loops 1192:1196 arg1 flexibility 1160:1170 the flexibility 1156:1170 the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition 1156:1265 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 7 11 theme preexisting 1351:1361 arg1 conformation 1370:1381 the preexisting glycan conformation 1347:1381 the preexisting glycan conformation 1347:1381 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 4 12 theme molecular 729:737 arg1 simulations 748:758 molecular dynamics simulations 729:758 molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 729:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 6 13 theme V3 1189:1190 arg1 loops 1192:1196 V3 loops 1189:1196 V3 loops 1189:1196 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 3 14 theme HIV-1 550:554 arg1 gp120 556:560 HIV-1 gp120 550:560 HIV-1 gp120 550:560 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 15 theme HIV-1 695:699 arg1 isolates 701:708 HIV-1 isolates 695:708 HIV-1 isolates 695:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 7 16 theme determining 1448:1458 arg1 loops 1467:1471 flexible complementarity determining region loops 1423:1471 flexible complementarity determining region loops that sample the bound conformations occasionally 1423:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 1 17 theme Many 88:91 arg1 antibodies 120:129 Many novel broadly neutralizing antibodies 88:129 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) 88:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 7 18 theme flexible 1313:1320 arg1 interface 1322:1330 the flexible interface 1309:1330 the flexible interface 1309:1330 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 3 19 theme isolates 701:708 arg1 % 690:690 more than 70% 678:690 more than 70% of HIV-1 isolates 678:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 19 theme isolates 701:708 arg1 isolates 701:708 HIV-1 isolates 695:708 HIV-1 isolates 695:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 2 20 theme Structural 300:309 arg1 studies 311:317 Structural studies 300:317 Structural studies 300:317 Structural studies reveal that the epitopes of some of these antibodies involve one or more crucial glycans, without which the binding is completely abolished. 26537503 6 21 theme V1/V2 1179:1183 arg1 flexibility 1160:1170 the flexibility 1156:1170 the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition 1156:1265 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 3 22 dep antibodies 591:600 arg1 PG9 602:604 PG9 602:604 PG9 (targeting V1/V2) 602:622 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 22 dep antibodies 591:600 arg1 V1/V2 617:621 targeting V1/V2 607:621 targeting V1/V2 607:621 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 22 dep antibodies 591:600 arg1 PGT128 628:633 PGT128 628:633 PGT128 (targeting V3) 628:648 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 22 dep antibodies 591:600 arg1 V3 646:647 targeting V3 636:647 targeting V3 636:647 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 22 dep antibodies 591:600 arg1 antibodies 591:600 two broadly neutralizing antibodies 566:600 two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates 566:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 0 23 theme glycans 9:15 arg1 Roles 0:4 Roles 0:4 Roles of glycans in interactions between gp120 and HIV 0:53 Roles of glycans in interactions between gp120 and HIV broadly neutralizing antibodies. 26537503 1 24 theme HIV-1 285:289 arg1 vaccine 291:297 an effective HIV-1 vaccine 272:297 an effective HIV-1 vaccine 272:297 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 5 25 theme multivalent 1004:1014 arg1 interactions 1016:1027 the multivalent interactions 1000:1027 the multivalent interactions between the antibody and gp120 1000:1058 The simulation results show that the complex structures are stabilized by the glycans, and the multivalent interactions between the antibody and gp120 promote cooperativities to further enhance the binding. 26537503 1 26 theme broadly 99:105 arg1 antibodies 120:129 Many novel broadly neutralizing antibodies 88:129 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) 88:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 1 27 theme vaccine 291:297 arg1 development 257:267 the development 253:267 the development of an effective HIV-1 vaccine 253:297 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 6 28 theme recognition 1255:1265 arg1 cost 1234:1237 the entropy cost 1222:1237 the entropy cost of the antibody recognition 1222:1265 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 7 29 theme bound 1489:1493 arg1 conformations 1495:1507 the bound conformations 1485:1507 the bound conformations 1485:1507 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 4 30 theme number 765:770 arg1 simulations 748:758 molecular dynamics simulations 729:758 molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 729:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 1 31 theme past 206:209 arg1 decade 211:216 the past decade 202:216 the past decade 202:216 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 6 32 theme antibody 1246:1253 arg1 recognition 1255:1265 the antibody recognition 1242:1265 the antibody recognition 1242:1265 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 3 33 theme neutralizing 578:589 arg1 PG9 602:604 PG9 602:604 PG9 (targeting V1/V2) 602:622 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 33 theme neutralizing 578:589 arg1 PGT128 628:633 PGT128 628:633 PGT128 (targeting V3) 628:648 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 33 theme neutralizing 578:589 arg1 antibodies 591:600 two broadly neutralizing antibodies 566:600 two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates 566:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 2 34 theme crucial 392:398 arg1 glycans 400:406 one or more crucial glycans 380:406 one or more crucial glycans 380:406 Structural studies reveal that the epitopes of some of these antibodies involve one or more crucial glycans, without which the binding is completely abolished. 26537503 1 35 theme effective 275:283 arg1 vaccine 291:297 an effective HIV-1 vaccine 272:297 an effective HIV-1 vaccine 272:297 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 4 36 theme systems 775:781 arg1 complex 808:814 antibody-gp120 complex 793:814 antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 793:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 4 36 theme systems 775:781 arg1 number 765:770 a number 763:770 a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 763:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 1 37 theme neutralizing 107:118 arg1 antibodies 120:129 Many novel broadly neutralizing antibodies 88:129 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) 88:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 7 38 theme shield 1411:1416 arg1 glycan 1404:1409 the glycan 1400:1409 the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally 1400:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 5 39 dep antibody 1041:1048 arg1 the 1037:1039 the 1037:1039 the 1037:1039 The simulation results show that the complex structures are stabilized by the glycans, and the multivalent interactions between the antibody and gp120 promote cooperativities to further enhance the binding. 26537503 7 40 theme region 1460:1465 arg1 loops 1467:1471 flexible complementarity determining region loops 1423:1471 flexible complementarity determining region loops that sample the bound conformations occasionally 1423:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 7 41 theme glycan 1363:1368 arg1 conformation 1370:1381 the preexisting glycan conformation 1347:1381 the preexisting glycan conformation 1347:1381 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 4 42 theme dynamics 739:746 arg1 simulations 748:758 molecular dynamics simulations 729:758 molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems 729:906 We have performed molecular dynamics simulations of a number of systems including antibody-gp120 complex with and without glycans, antibody, gp120 with and without glycans, and glycan-only systems. 26537503 1 43 theme novel 93:97 arg1 antibodies 120:129 Many novel broadly neutralizing antibodies 88:129 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) 88:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 6 44 theme entropy 1226:1232 arg1 cost 1234:1237 the entropy cost 1222:1237 the entropy cost of the antibody recognition 1222:1265 In the free gp120, the glycans increase the flexibility of the V1/V2 and V3 loops, which likely increases the entropy cost of the antibody recognition. 26537503 7 45 theme complementarity 1432:1446 arg1 loops 1467:1471 flexible complementarity determining region loops 1423:1471 flexible complementarity determining region loops that sample the bound conformations occasionally 1423:1520 However, the antibodies are able to bind the flexible interface by recognizing the preexisting glycan conformation, and penetrating the glycan shield with flexible complementarity determining region loops that sample the bound conformations occasionally. 26537503 3 46 theme targeting 607:615 arg1 PG9 602:604 PG9 602:604 PG9 (targeting V1/V2) 602:622 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 46 theme targeting 607:615 arg1 V1/V2 617:621 targeting V1/V2 607:621 targeting V1/V2 607:621 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 1 47 theme human 139:143 arg1 virus 162:166 human immunodeficiency virus 139:166 human immunodeficiency virus (HIV) 139:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 1 47 theme human 139:143 arg1 HIV 169:171 HIV 169:171 HIV 169:171 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 3 48 theme targeting 636:644 arg1 PGT128 628:633 PGT128 628:633 PGT128 (targeting V3) 628:648 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 48 theme targeting 636:644 arg1 V3 646:647 targeting V3 636:647 targeting V3 636:647 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 5 49 theme complex 946:952 arg1 structures 954:963 the complex structures 942:963 the complex structures 942:963 The simulation results show that the complex structures are stabilized by the glycans, and the multivalent interactions between the antibody and gp120 promote cooperativities to further enhance the binding. 26537503 1 50 theme immunodeficiency 145:160 arg1 virus 162:166 human immunodeficiency virus 139:166 human immunodeficiency virus (HIV) 139:172 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 1 50 theme immunodeficiency 145:160 arg1 HIV 169:171 HIV 169:171 HIV 169:171 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 1 51 theme promising 229:237 arg1 templates 239:247 promising templates 229:247 promising templates 229:247 Many novel broadly neutralizing antibodies against human immunodeficiency virus (HIV) have been identified during the past decade, providing promising templates for the development of an effective HIV-1 vaccine. 26537503 3 52 theme critical 500:507 arg1 roles 509:513 the critical roles 496:513 the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates 496:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26537503 3 53 from roles 509:513 arg1 interactions 529:540 interactions 529:540 interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates 529:708 In this study, we have investigated the critical roles of glycans in interactions between HIV-1 gp120 and two broadly neutralizing antibodies PG9 (targeting V1/V2) and PGT128 (targeting V3) that are able to neutralize more than 70% of HIV-1 isolates. 26503433 4 0 with sample 686:691 arg1 quantitation 729:740 quantitation 729:740 quantitation 729:740 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 4 0 with sample 686:691 arg1 reproducibility 709:723 excellent reproducibility 699:723 excellent reproducibility 699:723 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 4 1 theme excellent 699:707 arg1 reproducibility 709:723 excellent reproducibility 699:723 excellent reproducibility 699:723 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 5 2 theme fucosylated 824:834 arg1 peaks 836:840 the fucosylated peaks 820:840 the fucosylated peaks 820:840 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 0 3 theme Liver 87:91 arg1 Cirrhosis 93:101 Liver Cirrhosis 87:101 Liver Cirrhosis 87:101 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 0 4 from Method 10:15 arg1 Carcinoma 73:81 Hepatocellular Carcinoma 58:81 Hepatocellular Carcinoma 58:81 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 0 4 from Method 10:15 arg1 Cirrhosis 93:101 Liver Cirrhosis 87:101 Liver Cirrhosis 87:101 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 4 5 from separation 540:549 arg1 column 589:594 a HILIC column 581:594 a HILIC column 581:594 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 1 6 from haptoglobin 159:169 arg1 detection 121:129 the detection 117:129 the detection of fucosylated glycans from haptoglobin in patient serum 117:186 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 1 6 from haptoglobin 159:169 arg1 glycans 146:152 fucosylated glycans 134:152 fucosylated glycans from haptoglobin 134:169 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 2 7 theme HPLC-based 303:312 arg1 column 323:328 an HPLC-based affinity column 300:328 an HPLC-based affinity column followed by glycan removal, extraction, and desialylation 300:386 The workflow involves isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation. 26503433 5 8 theme samples 983:989 arg1 case 944:947 the case 940:947 the case 940:947 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 1 9 theme fucosylated 134:144 arg1 glycans 146:152 fucosylated glycans 134:152 fucosylated glycans from haptoglobin 134:169 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 1 10 theme patient 174:180 arg1 serum 182:186 patient serum 174:186 patient serum 174:186 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 5 11 theme good 1129:1132 arg1 marker 1134:1139 a particularly good marker 1114:1139 a particularly good marker for HCC 1114:1147 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 2 12 theme haptoglobin 282:292 arg1 isolation 265:273 isolation 265:273 isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation 265:386 The workflow involves isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation. 26503433 4 13 from haptoglobin 626:636 arg1 glycans 613:619 eight glycans 607:619 eight glycans from haptoglobin 607:636 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 3 14 gly fucosylated 393:403 arg1 glycans 405:411 The fucosylated glycans 389:411 The fucosylated glycans 389:411 The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. 26503433 5 15 theme hepatocellular 952:965 arg1 HCC 978:980 HCC 978:980 HCC 978:980 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 5 15 theme hepatocellular 952:965 arg1 carcinoma 967:975 hepatocellular carcinoma 952:975 hepatocellular carcinoma (HCC) samples 952:989 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 4 16 theme serum 680:684 arg1 sample 686:691 a serum sample 678:691 a serum sample 678:691 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 0 17 theme ESI-LC-MS 0:8 arg1 Method 10:15 ESI-LC-MS Method 0:15 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 0:102 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 4 18 theme HILIC 583:587 arg1 column 589:594 a HILIC column 581:594 a HILIC column 581:594 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 5 19 theme carcinoma 967:975 arg1 samples 983:989 hepatocellular carcinoma (HCC) samples 952:989 hepatocellular carcinoma (HCC) samples 952:989 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 0 20 theme Haptoglobin 21:31 arg1 Analysis 46:53 Haptoglobin Fucosylation Analysis 21:53 Haptoglobin Fucosylation Analysis 21:53 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 1 21 from detection 121:129 arg1 haptoglobin 159:169 haptoglobin 159:169 haptoglobin 159:169 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 1 21 from detection 121:129 arg1 serum 182:186 patient serum 174:186 patient serum 174:186 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 5 22 theme tetra-antennary 1066:1080 arg1 form 1082:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 3 23 theme fucosylated 393:403 arg1 glycans 405:411 The fucosylated glycans 389:411 The fucosylated glycans 389:411 The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. 26503433 2 24 theme affinity 314:321 arg1 column 323:328 an HPLC-based affinity column 300:328 an HPLC-based affinity column followed by glycan removal, extraction, and desialylation 300:386 The workflow involves isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation. 26503433 5 25 theme fucosylated 901:911 arg1 glycans 913:919 the fucosylated glycans 897:919 the fucosylated glycans 897:919 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 1 26 gly fucosylated 134:144 arg1 glycans 146:152 fucosylated glycans 134:152 fucosylated glycans from haptoglobin 134:169 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 5 27 theme corresponding 851:863 arg1 forms 880:884 their corresponding nonfucosylated forms 845:884 their corresponding nonfucosylated forms 845:884 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 0 28 theme Fucosylation 33:44 arg1 Analysis 46:53 Haptoglobin Fucosylation Analysis 21:53 Haptoglobin Fucosylation Analysis 21:53 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 4 29 theme derivatized 558:568 arg1 glycans 570:576 the derivatized glycans 554:576 the derivatized glycans 554:576 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 5 30 gly nonfucosylated 865:878 arg1 forms 880:884 their corresponding nonfucosylated forms 845:884 their corresponding nonfucosylated forms 845:884 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 5 31 gly bifucosylated 1052:1064 arg1 form 1082:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 1 32 theme enhanced 221:228 arg1 sensitivity 230:240 enhanced sensitivity 221:240 enhanced sensitivity 221:240 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 0 33 theme Hepatocellular 58:71 arg1 Carcinoma 73:81 Hepatocellular Carcinoma 58:81 Hepatocellular Carcinoma 58:81 ESI-LC-MS Method for Haptoglobin Fucosylation Analysis in Hepatocellular Carcinoma and Liver Cirrhosis. 26503433 5 34 theme abundance 1042:1050 arg1 form 1082:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 3 35 from detection 483:491 arg1 ionization 524:533 electrospray ionization 511:533 electrospray ionization 511:533 The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. 26503433 3 36 theme electrospray 511:522 arg1 ionization 524:533 electrospray ionization 511:533 electrospray ionization 511:533 The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. 26503433 5 37 gly fucosylated 901:911 arg1 glycans 913:919 the fucosylated glycans 897:919 the fucosylated glycans 897:919 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 5 38 theme bifucosylated 1052:1064 arg1 form 1082:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 5 39 gly fucosylated 824:834 arg1 peaks 836:840 the fucosylated peaks 820:840 the fucosylated peaks 820:840 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 4 40 theme glycans 570:576 arg1 separation 540:549 The separation 536:549 The separation of the derivatized glycans in a HILIC column 536:594 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 26503433 5 41 theme samples 1008:1014 arg1 case 944:947 the case 940:947 the case 940:947 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 5 42 theme low 1038:1040 arg1 abundance 1042:1050 the relatively low abundance 1023:1050 the relatively low abundance bifucosylated tetra-antennary form 1023:1085 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 3 43 theme glycans 500:506 arg1 detection 483:491 the detection 479:491 the detection of the glycans in electrospray ionization 479:533 The fucosylated glycans are then derivatized by Meladrazine, which significantly enhances the detection of the glycans in electrospray ionization. 26503433 5 44 theme cirrhosis 998:1006 arg1 samples 1008:1014 cirrhosis samples 998:1014 cirrhosis samples 998:1014 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 2 45 theme glycan 342:347 arg1 removal 349:355 glycan removal 342:355 glycan removal 342:355 The workflow involves isolation of the haptoglobin using an HPLC-based affinity column followed by glycan removal, extraction, and desialylation. 26503433 5 46 theme nonfucosylated 865:878 arg1 forms 880:884 their corresponding nonfucosylated forms 845:884 their corresponding nonfucosylated forms 845:884 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 1 47 theme glycans 146:152 arg1 detection 121:129 the detection 117:129 the detection of fucosylated glycans from haptoglobin in patient serum 117:186 A method for the detection of fucosylated glycans from haptoglobin in patient serum has been developed that provides enhanced sensitivity. 26503433 5 48 theme peaks 836:840 arg1 ratio 811:815 The ratio 807:815 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms 807:884 The ratio of the fucosylated peaks to their corresponding nonfucosylated forms shows that the fucosylated glycans are upregulated in the case of hepatocellular carcinoma (HCC) samples versus cirrhosis samples, where the relatively low abundance bifucosylated tetra-antennary form can be detected and may be a particularly good marker for HCC. 26503433 4 49 theme sample 686:691 arg1 μL 672:673 less than 1 μL 660:673 less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected 660:804 The separation of the derivatized glycans in a HILIC column shows that eight glycans from haptoglobin can be detected using less than 1 μL of a serum sample, with excellent reproducibility and quantitation, where without derivatization the glycans could not be detected. 25837443 6 0 theme certain 935:941 arg1 glycans 943:949 certain glycans 935:949 certain glycans 935:949 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 5 1 theme affinity 568:575 arg1 isolations 577:586 Lectin affinity isolations 561:586 Lectin affinity isolations 561:586 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 4 2 theme specific 543:550 arg1 glycans 552:558 specific glycans 543:558 specific glycans 543:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 3 3 gly glycoproteins 427:439 arg1 glycoproteins 427:439 their affected glycoproteins 412:439 their affected glycoproteins 412:439 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 5 4 theme captured 635:642 arg1 glycopeptides 644:656 the captured glycopeptides 631:656 the captured glycopeptides 631:656 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 4 5 theme novel 460:464 arg1 method 466:471 a novel method 458:471 a novel method for rapid and quantitative identification of glycoproteins containing specific glycans 458:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 3 6 gly heterogeneity 287:299 arg1 glycosylation 304:316 glycosylation 304:316 glycosylation 304:316 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 5 7 theme glycopeptides 644:656 arg1 immobilization 613:626 chemical immobilization 604:626 chemical immobilization of the captured glycopeptides 604:656 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 6 8 theme method 788:793 arg1 application 769:779 The application 765:779 The application of the method 765:793 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 8 theme method 788:793 arg1 discover 901:908 discover 901:908 to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets 898:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 8 theme method 788:793 arg1 useful 805:810 useful 805:810 useful 805:810 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 8 theme method 788:793 arg1 facilitate 815:824 facilitate 815:824 to facilitate our understanding of how changes in glycan associate with diseases 812:891 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 2 9 gly glycosylated 180:191 arg1 glycoprotein 160:171 Each glycoprotein 155:171 Each glycoprotein 155:171 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 2 9 gly glycosylated 180:191 arg2 glycosites 205:214 multiple glycosites 196:214 multiple glycosites 196:214 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 2 10 gly glycoprotein 160:171 arg1 glycoprotein 160:171 Each glycoprotein 155:171 Each glycoprotein 155:171 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 4 11 theme rapid 477:481 arg1 identification 500:513 rapid and quantitative identification 477:513 rapid and quantitative identification of glycoproteins containing specific glycans 477:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 4 12 gly glycoproteins 518:530 arg1 glycoproteins 518:530 glycoproteins 518:530 glycoproteins containing specific glycans 518:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 6 13 with glycoproteins 916:928 arg1 glycans 943:949 certain glycans 935:949 certain glycans 935:949 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 5 14 theme chemical 604:611 arg1 immobilization 613:626 chemical immobilization 604:626 chemical immobilization of the captured glycopeptides 604:656 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 6 15 theme therapeutic 985:995 arg1 glycoproteins 916:928 novel glycoproteins 910:928 novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets 910:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 15 theme therapeutic 985:995 arg1 targets 997:1003 therapeutic targets 985:1003 therapeutic targets 985:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 3 16 theme glycans 400:406 arg1 relationship 375:386 the structure-function relationship 352:386 the structure-function relationship of specific glycans and their affected glycoproteins 352:439 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 4 17 theme quantitative 487:498 arg1 identification 500:513 rapid and quantitative identification 477:513 rapid and quantitative identification of glycoproteins containing specific glycans 477:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 0 18 contain containing 32:41 arg1 glycoproteins 18:30 glycoproteins 18:30 glycoproteins containing specific glycans 18:58 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 0 18 contain containing 32:41 arg2 glycans 52:58 specific glycans 43:58 specific glycans 43:58 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 5 19 theme glycoproteins 690:702 arg1 identification 672:685 the identification 668:685 the identification of glycoproteins containing specific glycans by subsequent mass spectrometry 668:762 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 5 20 theme specific 715:722 arg1 glycans 724:730 specific glycans 715:730 specific glycans 715:730 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 3 21 theme specific 391:398 arg1 glycans 400:406 specific glycans 391:406 specific glycans 391:406 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 2 22 theme multiple 196:203 arg1 glycosites 205:214 multiple glycosites 196:214 multiple glycosites 196:214 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 3 23 theme glycoproteins 427:439 arg1 relationship 375:386 the structure-function relationship 352:386 the structure-function relationship of specific glycans and their affected glycoproteins 352:439 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 6 24 gly glycoproteins 916:928 arg1 glycoproteins 916:928 novel glycoproteins 910:928 novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets 910:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 24 gly glycoproteins 916:928 arg1 targets 997:1003 therapeutic targets 985:1003 therapeutic targets 985:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 24 gly glycoproteins 916:928 arg1 biomarkers 971:980 biomarkers 971:980 biomarkers 971:980 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 5 25 gly glycoproteins 690:702 arg1 glycoproteins 690:702 glycoproteins 690:702 glycoproteins containing specific glycans by subsequent mass spectrometry 690:762 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 0 26 theme specific 43:50 arg1 glycans 52:58 specific glycans 43:58 specific glycans 43:58 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 6 27 from changes 851:857 arg1 glycan 862:867 glycan 862:867 glycan 862:867 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 2 28 gly glycosites 226:235 arg2 glycosites 226:235 each glycosites 221:235 each glycosites 221:235 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 3 29 theme structure-function 356:373 arg1 relationship 375:386 the structure-function relationship 352:386 the structure-function relationship of specific glycans and their affected glycoproteins 352:439 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 5 30 theme subsequent 735:744 arg1 spectrometry 751:762 subsequent mass spectrometry 735:762 subsequent mass spectrometry 735:762 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 6 31 theme novel 910:914 arg1 glycoproteins 916:928 novel glycoproteins 910:928 novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets 910:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 31 theme novel 910:914 arg1 targets 997:1003 therapeutic targets 985:1003 therapeutic targets 985:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 31 theme novel 910:914 arg1 biomarkers 971:980 biomarkers 971:980 biomarkers 971:980 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 0 32 gly glycoproteins 18:30 arg1 glycoproteins 18:30 glycoproteins 18:30 glycoproteins containing specific glycans 18:58 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 3 33 theme affected 418:425 arg1 glycoproteins 427:439 their affected glycoproteins 412:439 their affected glycoproteins 412:439 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 5 34 theme mass 746:749 arg1 spectrometry 751:762 subsequent mass spectrometry 735:762 subsequent mass spectrometry 735:762 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 1 35 theme common 125:130 arg1 modifications 140:152 the most common protein modifications 116:152 the most common protein modifications 116:152 Glycosylation is one of the most common protein modifications. 25837443 6 36 dep useful 805:810 arg1 facilitate 815:824 facilitate 815:824 to facilitate our understanding of how changes in glycan associate with diseases 812:891 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 36 dep useful 805:810 arg1 discover 901:908 discover 901:908 to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets 898:1003 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 36 dep useful 805:810 arg1 useful 805:810 useful 805:810 useful 805:810 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 6 36 dep useful 805:810 arg1 application 769:779 The application 765:779 The application of the method 765:793 The application of the method should be useful to facilitate our understanding of how changes in glycan associate with diseases, and to discover novel glycoproteins with certain glycans that could serve as biomarkers or therapeutic targets. 25837443 2 37 mod modified 244:251 arg1 glycosites 226:235 each glycosites 221:235 each glycosites 221:235 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 2 37 mod modified 244:251 arg3 glycans 266:272 different glycans 256:272 different glycans 256:272 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 1 38 theme protein 132:138 arg1 modifications 140:152 the most common protein modifications 116:152 the most common protein modifications 116:152 Glycosylation is one of the most common protein modifications. 25837443 5 39 gly glycopeptides 644:656 arg2 glycopeptides 644:656 the captured glycopeptides 631:656 the captured glycopeptides 631:656 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 3 40 theme glycosylation 304:316 arg1 heterogeneity 287:299 this heterogeneity 282:299 this heterogeneity of glycosylation 282:316 Due to this heterogeneity of glycosylation, it has proven difficult to study the structure-function relationship of specific glycans and their affected glycoproteins. 25837443 1 41 theme modifications 140:152 arg1 one 109:111 one 109:111 one 109:111 Glycosylation is one of the most common protein modifications. 25837443 1 41 theme modifications 140:152 arg1 modifications 140:152 the most common protein modifications 116:152 the most common protein modifications 116:152 Glycosylation is one of the most common protein modifications. 25837443 2 42 gly glycosites 205:214 arg2 glycosites 205:214 multiple glycosites 196:214 multiple glycosites 196:214 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 2 43 theme different 256:264 arg1 glycans 266:272 different glycans 256:272 different glycans 256:272 Each glycoprotein can be glycosylated at multiple glycosites, and each glycosites can be modified by different glycans. 25837443 4 44 contain containing 532:541 arg1 glycoproteins 518:530 glycoproteins 518:530 glycoproteins containing specific glycans 518:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 4 44 contain containing 532:541 arg2 glycans 552:558 specific glycans 543:558 specific glycans 543:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 0 45 theme lectin-chemical 68:82 arg1 method 84:89 a lectin-chemical method 66:89 a lectin-chemical method 66:89 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 4 46 theme glycoproteins 518:530 arg1 identification 500:513 rapid and quantitative identification 477:513 rapid and quantitative identification of glycoproteins containing specific glycans 477:558 Here, we report a novel method for rapid and quantitative identification of glycoproteins containing specific glycans. 25837443 0 47 theme glycoproteins 18:30 arg1 Identification 0:13 Identification 0:13 Identification of glycoproteins containing specific glycans 0:58 Identification of glycoproteins containing specific glycans using a lectin-chemical method. 25837443 5 48 theme Lectin 561:566 arg1 isolations 577:586 Lectin affinity isolations 561:586 Lectin affinity isolations 561:586 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 5 49 contain containing 704:713 arg1 glycoproteins 690:702 glycoproteins 690:702 glycoproteins containing specific glycans by subsequent mass spectrometry 690:762 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 25837443 5 49 contain containing 704:713 arg2 glycans 724:730 specific glycans 715:730 specific glycans 715:730 Lectin affinity isolations are followed by chemical immobilization of the captured glycopeptides, allowing the identification of glycoproteins containing specific glycans by subsequent mass spectrometry. 28272765 0 0 theme 1,1'-Disaccharides 117:134 arg1 Synthesis 104:112 Synthesis 104:112 Synthesis 104:112 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 0 0 theme 1,1'-Disaccharides 117:134 arg1 1,1'-Glycosylation 81:98 Nonsymmetrical 1,1'-Glycosylation 66:98 Nonsymmetrical 1,1'-Glycosylation 66:98 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 4 1 theme class 609:613 arg1 forces 593:598 the joined forces 582:598 the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors 582:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 1 2 theme structural 234:243 arg1 components 245:254 structural components 234:254 structural components in various glycolipids and natural products 234:298 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 3 3 theme stereoselective 408:422 arg1 strategy 443:450 a direct yet stereoselective 1,1'-glycosylation strategy 395:450 a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components 395:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 4 4 theme stable 636:641 arg1 acceptors 653:661 configurationally stable glycoside acceptors 618:661 configurationally stable glycoside acceptors 618:661 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 5 5 theme remote 783:788 arg1 position 796:803 the remote C4/C3 position 779:803 the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions 779:876 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 0 6 theme Diverse 141:147 arg1 Configurations 149:162 Diverse Configurations 141:162 Diverse Configurations 141:162 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 5 7 theme C4/C3 790:794 arg1 position 796:803 the remote C4/C3 position 779:803 the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions 779:876 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 1 8 from components 245:254 arg1 glycolipids 267:277 various glycolipids 259:277 various glycolipids 259:277 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 1 8 from components 245:254 arg1 products 291:298 natural products 283:298 natural products 283:298 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 0 9 with Synthesis 104:112 arg1 Configurations 149:162 Diverse Configurations 141:162 Diverse Configurations 141:162 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 2 10 theme biological 362:371 arg1 properties 373:382 appealing biological properties 352:382 appealing biological properties 352:382 Some of these compounds have been shown to exhibit appealing biological properties. 28272765 4 11 theme new 605:607 arg1 class 609:613 a new class 603:613 a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors 603:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 5 12 theme new 709:711 arg1 acceptors 723:731 The new glycoside acceptors 705:731 The new glycoside acceptors 705:731 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 2 13 theme appealing 352:360 arg1 properties 373:382 appealing biological properties 352:382 appealing biological properties 352:382 Some of these compounds have been shown to exhibit appealing biological properties. 28272765 1 14 theme various 259:265 arg1 glycolipids 267:277 various glycolipids 259:277 various glycolipids 259:277 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 0 15 theme Stable 10:15 arg1 Glycosides 51:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides 0:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 0:163 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 5 16 theme glycoside 713:721 arg1 acceptors 723:731 The new glycoside acceptors 705:731 The new glycoside acceptors 705:731 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 3 17 theme diverse 512:518 arg1 configurations 520:533 diverse configurations 512:533 diverse configurations 512:533 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 4 18 theme donors 697:702 arg1 class 609:613 a new class 603:613 a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors 603:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 3 19 with 1,1'-disaccharides 488:505 arg1 configurations 520:533 diverse configurations 512:533 diverse configurations 512:533 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 3 19 with 1,1'-disaccharides 488:505 arg1 components 545:554 sugar components 539:554 sugar components 539:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 5 20 theme acidic 860:865 arg1 conditions 867:876 acidic conditions 860:876 acidic conditions 860:876 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 0 21 theme Trimethylsilyl 36:49 arg1 Glycosides 51:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides 0:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 0:163 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 4 22 theme thioglycoside 683:695 arg1 donors 697:702 stereodirecting thioglycoside donors 667:702 stereodirecting thioglycoside donors 667:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 1 23 theme natural 283:289 arg1 products 291:298 natural products 283:298 natural products 283:298 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 3 24 theme 1,1'-disaccharides 488:505 arg1 synthesis 460:468 the synthesis 456:468 the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components 456:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 5 25 theme TMS 839:841 arg1 glycosides 843:852 TMS glycosides 839:852 TMS glycosides 839:852 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 3 26 theme sugar 539:543 arg1 components 545:554 sugar components 539:554 sugar components 539:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 0 27 theme Picoloyl-Protected 17:34 arg1 Glycosides 51:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides 0:60 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 0:163 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 4 28 theme acceptors 653:661 arg1 class 609:613 a new class 603:613 a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors 603:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 4 29 theme joined 586:591 arg1 forces 593:598 the joined forces 582:598 the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors 582:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 3 30 theme nonsymmetrical 473:486 arg1 1,1'-disaccharides 488:505 nonsymmetrical 1,1'-disaccharides 473:505 nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components 473:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 4 31 theme glycoside 643:651 arg1 acceptors 653:661 configurationally stable glycoside acceptors 618:661 configurationally stable glycoside acceptors 618:661 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 1 32 theme Nonsymmetrical 165:178 arg1 1,1'-disaccharides 180:197 Nonsymmetrical 1,1'-disaccharides 165:197 Nonsymmetrical 1,1'-disaccharides 165:197 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 0 33 theme Nonsymmetrical 66:79 arg1 1,1'-Glycosylation 81:98 Nonsymmetrical 1,1'-Glycosylation 66:98 Nonsymmetrical 1,1'-Glycosylation 66:98 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 4 34 theme stereodirecting 667:681 arg1 donors 697:702 stereodirecting thioglycoside donors 667:702 stereodirecting thioglycoside donors 667:702 The strategy is based on the joined forces of a new class of configurationally stable glycoside acceptors and stereodirecting thioglycoside donors. 28272765 3 35 theme direct 397:402 arg1 strategy 443:450 a direct yet stereoselective 1,1'-glycosylation strategy 395:450 a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components 395:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 5 36 theme unusual 818:824 arg1 stability 826:834 unusual stability 818:834 unusual stability 818:834 The new glycoside acceptors feature a picoloyl (Pico) protecting group at the remote C4/C3 position that confers unusual stability on TMS glycosides under acidic conditions. 28272765 0 37 with 1,1'-Glycosylation 81:98 arg1 Configurations 149:162 Diverse Configurations 141:162 Diverse Configurations 141:162 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 28272765 1 38 theme related 203:209 arg1 derivatives 211:221 related derivatives 203:221 related derivatives 203:221 Nonsymmetrical 1,1'-disaccharides and related derivatives constitute structural components in various glycolipids and natural products. 28272765 3 39 theme 1,1'-glycosylation 424:441 arg1 strategy 443:450 a direct yet stereoselective 1,1'-glycosylation strategy 395:450 a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components 395:554 We report a direct yet stereoselective 1,1'-glycosylation strategy for the synthesis of nonsymmetrical 1,1'-disaccharides with diverse configurations and sugar components. 28272765 0 40 gly 1,1'-Glycosylation 81:98 arg1 1,1'-Disaccharides 117:134 1,1'-Disaccharides 117:134 1,1'-Disaccharides 117:134 Unusually Stable Picoloyl-Protected Trimethylsilyl Glycosides for Nonsymmetrical 1,1'-Glycosylation and Synthesis of 1,1'-Disaccharides with Diverse Configurations. 26383616 1 0 theme other 154:158 arg1 biopharmaceuticals 173:190 other glycoprotein biopharmaceuticals 154:190 other glycoprotein biopharmaceuticals 154:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 1 1 theme glycoprotein 160:171 arg1 biopharmaceuticals 173:190 other glycoprotein biopharmaceuticals 154:190 other glycoprotein biopharmaceuticals 154:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 3 2 theme interest 629:636 arg1 components 615:624 all sample components 604:624 all sample components of interest 604:636 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 4 3 theme N-glycans 781:789 arg1 GUs 749:751 CE GUs 746:751 CE GUs 746:751 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 4 3 theme N-glycans 781:789 arg1 structures 767:776 suggested structures 757:776 suggested structures of N-glycans released from human IgG 757:813 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 3 4 theme unit 583:586 arg1 values 593:598 the glucose unit (GU) values 571:598 the glucose unit (GU) values for all sample components of interest 571:636 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 2 5 theme choice 353:358 arg1 methods 342:348 the high-performance methods 321:348 the high-performance methods of choice 321:358 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 2 6 theme necessary 380:388 arg1 instrumentation 390:404 the necessary instrumentation 376:404 the necessary instrumentation 376:404 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 1 7 theme biopharmaceuticals 173:190 arg1 use 110:112 the use 106:112 the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals 106:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 4 8 theme human 805:809 arg1 IgG 811:813 human IgG 805:813 human IgG 805:813 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 1 9 theme Recent 86:91 arg1 emergence 93:101 Recent emergence 86:101 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals 86:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 1 10 theme high-throughput 201:215 arg1 techniques 240:249 high-throughput, robust, and automated techniques 201:249 high-throughput, robust, and automated techniques 201:249 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 3 11 theme concomitant 668:678 arg1 search 689:694 a concomitant database search 666:694 a concomitant database search for structural assignment 666:720 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 3 12 theme GU 589:590 arg1 values 593:598 the glucose unit (GU) values 571:598 the glucose unit (GU) values for all sample components of interest 571:636 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 5 13 theme CE-based 910:917 arg1 analysis 926:933 CE-based glycan analysis 910:933 CE-based glycan analysis 910:933 The app is freely available online (www.lendulet.uni-pannon.hu/gucal) and readily facilitates CE-based glycan analysis. 26383616 0 14 theme integrated 10:19 arg1 application 21:31 An integrated application 7:31 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 0:84 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 3 15 theme structural 700:709 arg1 assignment 711:720 structural assignment 700:720 structural assignment 700:720 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 2 16 theme methods 342:348 arg1 methods 342:348 the high-performance methods 321:348 the high-performance methods of choice 321:358 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 2 16 theme methods 342:348 arg1 one 314:316 one 314:316 one 314:316 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 1 17 theme robust 218:223 arg1 techniques 240:249 high-throughput, robust, and automated techniques 201:249 high-throughput, robust, and automated techniques 201:249 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 5 18 theme available 834:842 arg1 www.lendulet.uni-pannon.hu/gucal 852:883 www.lendulet.uni-pannon.hu/gucal 852:883 www.lendulet.uni-pannon.hu/gucal 852:883 The app is freely available online (www.lendulet.uni-pannon.hu/gucal) and readily facilitates CE-based glycan analysis. 26383616 5 18 theme available 834:842 arg1 online 844:849 freely available online 827:849 freely available online (www.lendulet.uni-pannon.hu/gucal) 827:884 The app is freely available online (www.lendulet.uni-pannon.hu/gucal) and readily facilitates CE-based glycan analysis. 26383616 5 18 theme available 834:842 arg1 app 820:822 The app 816:822 The app 816:822 The app is freely available online (www.lendulet.uni-pannon.hu/gucal) and readily facilitates CE-based glycan analysis. 26383616 4 19 attach released 791:798 arg2 N-glycans 781:789 N-glycans 781:789 N-glycans released from human IgG 781:813 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 4 19 attach released 791:798 arg1 IgG 811:813 human IgG 805:813 human IgG 805:813 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 2 20 theme high-performance 325:340 arg1 methods 342:348 the high-performance methods 321:348 the high-performance methods of choice 321:358 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 3 21 with electropherogram 644:659 arg1 search 689:694 a concomitant database search 666:694 a concomitant database search for structural assignment 666:720 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 5 22 theme glycan 919:924 arg1 analysis 926:933 CE-based glycan analysis 910:933 CE-based glycan analysis 910:933 The app is freely available online (www.lendulet.uni-pannon.hu/gucal) and readily facilitates CE-based glycan analysis. 26383616 3 23 theme dubbed 527:532 arg1 toolset 519:525 an integrated toolset 505:525 an integrated toolset dubbed as GUcal 505:541 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 2 24 theme bioinformatics 437:450 arg1 tools 452:456 the related bioinformatics tools 425:456 the related bioinformatics tools 425:456 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 1 25 theme automated 230:238 arg1 techniques 240:249 high-throughput, robust, and automated techniques 201:249 high-throughput, robust, and automated techniques 201:249 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 0 26 theme electrophoresis 47:61 arg1 analysis 76:83 capillary electrophoresis based glycan analysis 37:83 capillary electrophoresis based glycan analysis 37:83 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 3 27 theme integrated 508:517 arg1 toolset 519:525 an integrated toolset 505:525 an integrated toolset dubbed as GUcal 505:541 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 0 28 theme capillary 37:45 arg1 analysis 76:83 capillary electrophoresis based glycan analysis 37:83 capillary electrophoresis based glycan analysis 37:83 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 3 29 theme database 680:687 arg1 search 689:694 a concomitant database search 666:694 a concomitant database search for structural assignment 666:720 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 1 30 gly glycoprotein 160:171 arg1 glycoprotein 160:171 other glycoprotein biopharmaceuticals 154:190 other glycoprotein biopharmaceuticals 154:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 1 31 theme monoclonal 117:126 arg1 therapeutics 137:148 monoclonal antibody therapeutics 117:148 monoclonal antibody therapeutics 117:148 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 3 32 theme glucose 575:581 arg1 values 593:598 the glucose unit (GU) values 571:598 the glucose unit (GU) values for all sample components of interest 571:636 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 2 33 theme Capillary 285:293 arg1 electrophoresis 295:309 Capillary electrophoresis 285:309 Capillary electrophoresis 285:309 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 4 34 theme suggested 757:765 arg1 structures 767:776 suggested structures 757:776 suggested structures of N-glycans released from human IgG 757:813 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 1 35 theme antibody 128:135 arg1 therapeutics 137:148 monoclonal antibody therapeutics 117:148 monoclonal antibody therapeutics 117:148 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 4 36 theme CE 746:747 arg1 GUs 749:751 CE GUs 746:751 CE GUs 746:751 The database comprises CE GUs and suggested structures of N-glycans released from human IgG. 26383616 0 37 theme glycan 69:74 arg1 analysis 76:83 capillary electrophoresis based glycan analysis 37:83 capillary electrophoresis based glycan analysis 37:83 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 2 38 theme related 429:435 arg1 tools 452:456 the related bioinformatics tools 425:456 the related bioinformatics tools 425:456 Capillary electrophoresis is one of the high-performance methods of choice; however, while the necessary instrumentation is well developed, the related bioinformatics tools are lacked behind. 26383616 3 39 theme sample 608:613 arg1 components 615:624 all sample components 604:624 all sample components of interest 604:636 In this paper, we introduce an integrated toolset dubbed as GUcal, to automatically calculate the glucose unit (GU) values for all sample components of interest in an electropherogram with a concomitant database search for structural assignment. 26383616 1 40 theme therapeutics 137:148 arg1 use 110:112 the use 106:112 the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals 106:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 1 41 from emergence 93:101 arg1 use 110:112 the use 106:112 the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals 106:190 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26383616 0 42 theme based 63:67 arg1 analysis 76:83 capillary electrophoresis based glycan analysis 37:83 capillary electrophoresis based glycan analysis 37:83 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 0 43 dep GUcal 0:4 arg1 application 21:31 An integrated application 7:31 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 0:84 GUcal: An integrated application for capillary electrophoresis based glycan analysis. 26383616 1 44 theme glycosylation 261:273 arg1 analysis 275:282 their glycosylation analysis 255:282 their glycosylation analysis 255:282 Recent emergence in the use of monoclonal antibody therapeutics and other glycoprotein biopharmaceuticals requires high-throughput, robust, and automated techniques for their glycosylation analysis. 26482026 8 0 theme immobilization 1181:1194 arg1 consequence 1132:1142 the biophysical consequence 1116:1142 the biophysical consequence of protein glycosylation and protein immobilization 1116:1194 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 5 1 gly glycosylation 816:828 arg1 GO 852:853 GO 852:853 GO 852:853 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 2 from glycosylation 816:828 arg1 GO 852:853 GO 852:853 GO 852:853 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 2 3 dep provide 266:272 arg1 implemented 350:360 implemented 350:360 could be implemented when GO nanosheets is used as protein immobilization matrix 341:420 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 9 4 theme structural 1376:1385 arg1 dynamics 1387:1394 structural dynamics 1376:1394 structural dynamics that resulted in an increase in thermostability 1376:1442 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 0 5 theme biophysical 57:67 arg1 properties 69:78 protein biophysical properties 49:78 protein biophysical properties 49:78 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 1 6 dep BACKGROUND 81:90 arg1 provides 103:110 provides 103:110 provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability 103:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 5 7 theme catalysis 879:887 arg1 rate 864:867 the rate 860:867 the rate of enzyme catalysis 860:887 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 7 8 theme slight 1054:1059 arg1 changes 1061:1067 slight changes 1054:1067 slight changes in the secondary structure 1054:1094 However, GO produced slight changes in the secondary structure. 26482026 3 9 theme native 531:536 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 9 theme native 531:536 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 9 theme native 531:536 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 10 theme bilirubin 555:563 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 10 theme bilirubin 555:563 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 10 theme bilirubin 555:563 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 11 dep strategies 485:494 arg1 adsorbing 497:505 adsorbing 497:505 adsorbing 497:505 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 11 dep strategies 485:494 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 11 dep strategies 485:494 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 11 dep strategies 485:494 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 0 12 from influence 36:44 arg1 properties 69:78 protein biophysical properties 49:78 protein biophysical properties 49:78 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 6 13 gly glycosylation 956:968 arg1 BOD 1028:1030 BOD 1028:1030 BOD 1028:1030 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 8 14 theme thermal 1288:1294 arg1 inactivation 1296:1307 thermal inactivation 1288:1307 thermal inactivation 1288:1307 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 1 15 theme thermal 244:250 arg1 stability 252:260 thermal stability 244:260 thermal stability 244:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 4 16 theme spectroscopy 680:691 arg1 measurements 693:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 1 17 from influence 143:151 arg1 activity 220:227 protein catalytic activity 202:227 protein catalytic activity 202:227 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 1 17 from influence 143:151 arg1 stability 252:260 thermal stability 244:260 thermal stability 244:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 1 17 from influence 143:151 arg1 dynamics 230:237 dynamics 230:237 dynamics 230:237 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 10 18 theme protein 1611:1617 arg1 dynamics 1630:1637 protein structural dynamics 1611:1637 protein structural dynamics 1611:1637 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 1 19 theme oxide 165:169 arg1 influence 143:151 the influence 139:151 the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability 139:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 1 20 from information 124:134 arg1 influence 143:151 the influence 139:151 the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability 139:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 3 21 theme attaching 521:529 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 21 theme attaching 521:529 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 21 theme attaching 521:529 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 6 22 theme BOD 1028:1030 arg1 tertiary 989:996 the tertiary 985:996 the tertiary 985:996 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 6 22 theme BOD 1028:1030 arg1 structure 1015:1023 the secondary structure 1001:1023 the secondary structure of BOD 1001:1030 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 8 23 theme biophysical 1120:1130 arg1 consequence 1132:1142 the biophysical consequence 1116:1142 the biophysical consequence of protein glycosylation and protein immobilization 1116:1194 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 6 24 theme Structural 902:911 arg1 analysis 913:920 Structural analysis 902:920 Structural analysis by circular dichroism 902:942 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 10 25 theme chemical 1576:1583 arg1 glycosylation 1585:1597 chemical glycosylation 1576:1597 chemical glycosylation 1576:1597 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 2 26 theme stabilization 294:306 arg1 evidence 274:281 evidence 274:281 evidence of protein stabilization by glycans 274:317 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 0 27 theme Graphene 0:7 arg1 oxide 9:13 Graphene oxide 0:13 Graphene oxide as a protein matrix: influence on protein biophysical properties. 0:79 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 6 28 theme circular 925:932 arg1 dichroism 934:942 circular dichroism 925:942 circular dichroism 925:942 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 4 29 theme zeta-potential 640:653 arg1 measurements 693:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 2 30 theme protein 286:292 arg1 stabilization 294:306 protein stabilization 286:306 protein stabilization by glycans 286:317 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 5 31 from immobilization 834:847 arg1 GO 852:853 GO 852:853 GO 852:853 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 32 theme substrate 769:777 arg1 affinity 787:794 the substrate binding affinity 765:794 the substrate binding affinity 765:794 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 2 33 dep nanosheets 370:379 arg1 GO 367:368 GO 367:368 GO 367:368 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 10 34 theme glycosylation 1585:1597 arg1 usefulness 1562:1571 the usefulness 1558:1571 the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix 1558:1694 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 5 35 theme enzyme 872:877 arg1 catalysis 879:887 enzyme catalysis 872:887 enzyme catalysis 872:887 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 3 36 theme glycosylated 542:553 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 36 theme glycosylated 542:553 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 36 theme glycosylated 542:553 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 2 37 theme protein 392:398 arg1 nanosheets 370:379 GO nanosheets 367:379 GO nanosheets 367:379 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 2 37 theme protein 392:398 arg1 matrix 415:420 protein immobilization matrix 392:420 protein immobilization matrix 392:420 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 2 38 theme immobilization 400:413 arg1 nanosheets 370:379 GO nanosheets 367:379 GO nanosheets 367:379 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 2 38 theme immobilization 400:413 arg1 matrix 415:420 protein immobilization matrix 392:420 protein immobilization matrix 392:420 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 4 39 theme RESULTS 586:592 arg1 formation 607:615 RESULTS Bioconjugate formation 586:615 RESULTS Bioconjugate formation 586:615 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 9 40 from decrease 1450:1457 arg1 dynamics 1387:1394 structural dynamics 1376:1394 structural dynamics that resulted in an increase in thermostability 1376:1442 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 9 40 from decrease 1450:1457 arg1 activity 1476:1483 the catalytic activity 1462:1483 the catalytic activity for both, glycoconjugate and immobilized enzyme 1462:1531 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 1 41 theme graphene 156:163 arg1 oxide 165:169 graphene oxide 156:169 graphene oxide (GO) 156:174 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 10 42 theme structural 1619:1628 arg1 dynamics 1630:1637 protein structural dynamics 1611:1637 protein structural dynamics 1611:1637 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 3 43 theme different 475:483 arg1 adsorbing 497:505 adsorbing 497:505 adsorbing 497:505 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 43 theme different 475:483 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 43 theme different 475:483 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 5 44 theme kinetic 714:720 arg1 k 743:743 k 743:743 k(cat) 743:748 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 44 theme kinetic 714:720 arg1 k 734:734 k 734:734 k(m) 734:737 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 44 theme kinetic 714:720 arg1 parameters 722:731 Enzyme kinetic parameters 707:731 Enzyme kinetic parameters (k(m) and k(cat)) 707:749 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 4 45 theme FT-IR 633:637 arg1 measurements 693:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements 633:704 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 10 46 theme stable 1671:1676 arg1 matrix 1689:1694 a more stable GO-protein matrix 1664:1694 a more stable GO-protein matrix 1664:1694 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 1 47 theme glycans 191:197 arg1 influence 143:151 the influence 139:151 the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability 139:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 0 48 theme protein 20:26 arg1 matrix 28:33 a protein matrix 18:33 a protein matrix 18:33 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 8 49 theme glycosylation 1155:1167 arg1 consequence 1132:1142 the biophysical consequence 1116:1142 the biophysical consequence of protein glycosylation and protein immobilization 1116:1194 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 8 50 theme protein 1236:1242 arg1 changes 1254:1260 structural protein dynamical changes 1225:1260 structural protein dynamical changes 1225:1260 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 8 51 theme protein 1147:1153 arg1 glycosylation 1155:1167 protein glycosylation 1147:1167 protein glycosylation 1147:1167 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 8 52 theme structural 1225:1234 arg1 changes 1254:1260 structural protein dynamical changes 1225:1260 structural protein dynamical changes 1225:1260 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 7 53 theme secondary 1076:1084 arg1 structure 1086:1094 the secondary structure 1072:1094 the secondary structure 1072:1094 However, GO produced slight changes in the secondary structure. 26482026 7 54 from changes 1061:1067 arg1 structure 1086:1094 the secondary structure 1072:1094 the secondary structure 1072:1094 However, GO produced slight changes in the secondary structure. 26482026 9 55 from reduction 1363:1371 arg1 dynamics 1387:1394 structural dynamics 1376:1394 structural dynamics that resulted in an increase in thermostability 1376:1442 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 9 55 from reduction 1363:1371 arg1 activity 1476:1483 the catalytic activity 1462:1483 the catalytic activity for both, glycoconjugate and immobilized enzyme 1462:1531 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 1 56 dep oxide 165:169 arg1 GO 172:173 GO 172:173 GO 172:173 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 1 56 dep oxide 165:169 arg1 nanosheets 176:185 nanosheets 176:185 nanosheets 176:185 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 2 57 used used 384:387 arg2 nanosheets 370:379 GO nanosheets 367:379 GO nanosheets 367:379 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 2 57 used used 384:387 arg2 matrix 415:420 protein immobilization matrix 392:420 protein immobilization matrix 392:420 We provide evidence of protein stabilization by glycans and how this strategy could be implemented when GO nanosheets is used as protein immobilization matrix. 26482026 8 58 theme H/D 1271:1273 arg1 exchange 1275:1282 FT-IR H/D exchange 1265:1282 FT-IR H/D exchange 1265:1282 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 8 59 theme protein 1173:1179 arg1 immobilization 1181:1194 protein immobilization 1173:1194 protein immobilization 1173:1194 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 9 60 theme catalytic 1466:1474 arg1 activity 1476:1483 the catalytic activity 1462:1483 the catalytic activity for both, glycoconjugate and immobilized enzyme 1462:1531 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 1 61 theme fundamental 112:122 arg1 information 124:134 fundamental information 112:134 fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability 112:260 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 8 62 dep nanosheets 1202:1211 arg1 GO 1199:1200 GO 1199:1200 GO 1199:1200 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 10 63 theme GO-protein 1678:1687 arg1 matrix 1689:1694 a more stable GO-protein matrix 1664:1694 a more stable GO-protein matrix 1664:1694 These results establish the usefulness of chemical glycosylation to modulate protein structural dynamics and stability to develop a more stable GO-protein matrix. 26482026 8 64 theme FT-IR 1265:1269 arg1 exchange 1275:1282 FT-IR H/D exchange 1265:1282 FT-IR H/D exchange 1265:1282 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 1 65 theme protein 202:208 arg1 activity 220:227 protein catalytic activity 202:227 protein catalytic activity 202:227 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 6 66 theme secondary 1005:1013 arg1 structure 1015:1023 the secondary structure 1001:1023 the secondary structure of BOD 1001:1030 Structural analysis by circular dichroism showed that glycosylation did not affect the tertiary or the secondary structure of BOD. 26482026 9 67 from increase 1416:1423 arg1 thermostability 1428:1442 thermostability 1428:1442 thermostability 1428:1442 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 3 68 gly glycosylated 542:553 arg1 BOD 574:576 BOD 574:576 BOD 574:576 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 68 gly glycosylated 542:553 arg1 oxidase 565:571 covalently attaching native and glycosylated bilirubin oxidase 510:571 covalently attaching native and glycosylated bilirubin oxidase (BOD) 510:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 3 68 gly glycosylated 542:553 arg1 strategies 485:494 two different strategies 471:494 two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) 471:577 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 5 69 theme Enzyme 707:712 arg1 k 743:743 k 743:743 k(cat) 743:748 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 69 theme Enzyme 707:712 arg1 k 734:734 k 734:734 k(m) 734:737 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 69 theme Enzyme 707:712 arg1 parameters 722:731 Enzyme kinetic parameters 707:731 Enzyme kinetic parameters (k(m) and k(cat)) 707:749 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 1 70 theme catalytic 210:218 arg1 activity 220:227 protein catalytic activity 202:227 protein catalytic activity 202:227 BACKGROUND This study provides fundamental information on the influence of graphene oxide (GO) nanosheets and glycans on protein catalytic activity, dynamics, and thermal stability. 26482026 0 71 theme protein 49:55 arg1 properties 69:78 protein biophysical properties 49:78 protein biophysical properties 49:78 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 0 72 dep oxide 9:13 arg1 influence 36:44 influence 36:44 Graphene oxide as a protein matrix: influence on protein biophysical properties. 0:79 Graphene oxide as a protein matrix: influence on protein biophysical properties. 26482026 3 73 theme bioconjugates 435:447 arg1 series 425:430 A series 423:430 A series of bioconjugates 423:447 A series of bioconjugates was constructed using two different strategies: adsorbing or covalently attaching native and glycosylated bilirubin oxidase (BOD) to GO. 26482026 4 74 theme photoelectron 666:678 arg1 spectroscopy 680:691 X-ray photoelectron spectroscopy 660:691 X-ray photoelectron spectroscopy 660:691 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 9 75 theme immobilized 1514:1524 arg1 enzyme 1526:1531 immobilized enzyme 1514:1531 immobilized enzyme 1514:1531 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 5 76 dep parameters 722:731 arg1 cat 745:747 cat 745:747 cat 745:747 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 76 dep parameters 722:731 arg1 m 736:736 m 736:736 m 736:736 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 76 dep parameters 722:731 arg1 k 743:743 k 743:743 k(cat) 743:748 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 76 dep parameters 722:731 arg1 k 734:734 k 734:734 k(m) 734:737 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 5 76 dep parameters 722:731 arg1 parameters 722:731 Enzyme kinetic parameters 707:731 Enzyme kinetic parameters (k(m) and k(cat)) 707:749 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 9 77 dep CONCLUSIONS 1310:1320 arg1 found 1329:1333 found 1329:1333 was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme 1325:1531 CONCLUSIONS It was found that glycosylation caused a reduction in structural dynamics that resulted in an increase in thermostability and a decrease in the catalytic activity for both, glycoconjugate and immobilized enzyme. 26482026 5 78 theme binding 779:785 arg1 affinity 787:794 the substrate binding affinity 765:794 the substrate binding affinity 765:794 Enzyme kinetic parameters (k(m) and k(cat)) revealed that the substrate binding affinity was not affected by glycosylation and immobilization on GO, but the rate of enzyme catalysis was reduced. 26482026 4 79 theme X-ray 660:664 arg1 spectroscopy 680:691 X-ray photoelectron spectroscopy 660:691 X-ray photoelectron spectroscopy 660:691 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 26482026 8 80 theme dynamical 1244:1252 arg1 changes 1254:1260 structural protein dynamical changes 1225:1260 structural protein dynamical changes 1225:1260 To shed light into the biophysical consequence of protein glycosylation and protein immobilization on GO nanosheets, we studied structural protein dynamical changes by FT-IR H/D exchange and thermal inactivation. 26482026 4 81 theme Bioconjugate 594:605 arg1 formation 607:615 RESULTS Bioconjugate formation 586:615 RESULTS Bioconjugate formation 586:615 RESULTS Bioconjugate formation was followed by FT-IR, zeta-potential, and X-ray photoelectron spectroscopy measurements. 27159528 5 0 theme ER- 722:724 arg1 Accumulation 706:717 Accumulation 706:717 Accumulation of ER- and vac-Abs 706:736 Accumulation of ER- and vac-Abs was 10- to 15-fold higher than sec-Ab. 27159528 6 1 theme GnGnXF 883:888 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 9 2 dep pathways 1302:1309 arg1 transport 1319:1327 direct transport 1312:1327 direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) 1312:1400 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 2 dep pathways 1302:1309 arg1 trafficking 1406:1416 trafficking 1406:1416 trafficking 1406:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 2 dep pathways 1302:1309 arg1 pathways 1302:1309 two different pathways 1288:1309 two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking 1288:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 8 3 theme microscopy 1080:1089 arg1 analysis 1091:1098 Confocal microscopy analysis 1071:1098 Confocal microscopy analysis using RFP fusions 1071:1116 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 1 4 theme monoclonal 168:177 arg1 antibodies 179:188 monoclonal antibodies 168:188 monoclonal antibodies 168:188 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 11 5 theme efficient 1641:1649 arg1 production 1651:1660 the efficient production 1637:1660 the efficient production of Abs with appropriate post-translational modifications 1637:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 11 6 theme current 1759:1765 arg1 concepts 1767:1774 current concepts 1759:1774 current concepts 1759:1774 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 0 7 from targeting 9:17 arg1 benthamiana 58:68 Nicotiana benthamiana 48:68 Nicotiana benthamiana 48:68 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana. 27159528 11 8 theme post-translational 1686:1703 arg1 modifications 1705:1717 appropriate post-translational modifications 1674:1717 appropriate post-translational modifications 1674:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 4 9 theme ER-retained 654:664 arg1 variants 666:673 ER-retained variants 654:673 ER-retained variants 654:673 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 9 10 theme Man 1386:1388 arg1 structures 1390:1399 Man structures 1386:1399 Man structures 1386:1399 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 3 11 from targeting 410:418 arg1 benthamiana 485:495 Nicotiana benthamiana 475:495 Nicotiana benthamiana leaves 475:502 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 5 12 dep 15-fold 749:755 arg1 higher 757:762 higher 757:762 higher 757:762 Accumulation of ER- and vac-Abs was 10- to 15-fold higher than sec-Ab. 27159528 5 12 dep 15-fold 749:755 arg1 to 746:747 to 746:747 to 746:747 Accumulation of ER- and vac-Abs was 10- to 15-fold higher than sec-Ab. 27159528 7 13 dep glycans 1004:1010 arg1 assigned 1022:1029 assigned 1022:1029 assigned as typical vacuolar 1022:1049 Paucimannosidic glycans (commonly assigned as typical vacuolar) were not detected. 27159528 6 14 theme fucosylated 855:865 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 9 15 from ER 1338:1339 arg1 transport 1319:1327 direct transport 1312:1327 direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) 1312:1400 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 15 from ER 1338:1339 arg1 trafficking 1406:1416 trafficking 1406:1416 trafficking 1406:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 15 from ER 1338:1339 arg1 pathways 1302:1309 two different pathways 1288:1309 two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking 1288:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 1 16 theme Plant-based 71:81 arg1 platforms 83:91 Plant-based platforms 71:91 Plant-based platforms 71:91 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 9 17 dep Golgi 1355:1359 arg1 molecules 1365:1373 Ab molecules 1362:1373 Ab molecules containing Man structures 1362:1399 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 1 18 theme proteins 148:155 arg1 expression 122:131 the expression 118:131 the expression of recombinant proteins, including monoclonal antibodies 118:188 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 6 19 theme complex 847:853 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 2 20 theme better 277:282 arg1 understanding 284:296 a better understanding 275:296 a better understanding of intracellular processes that affect protein properties 275:354 However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. 27159528 8 21 theme central 1218:1224 arg1 vacuole 1226:1232 the central vacuole 1214:1232 the central vacuole 1214:1232 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 0 22 theme Nicotiana 48:56 arg1 benthamiana 58:68 Nicotiana benthamiana 48:68 Nicotiana benthamiana 48:68 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana. 27159528 6 23 theme typical 839:845 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 0 24 theme Vacuolar 0:7 arg1 targeting 9:17 Vacuolar targeting 0:17 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana 0:68 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana. 27159528 11 25 theme appropriate 1609:1619 arg1 compartment 1621:1631 an appropriate compartment 1606:1631 an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications 1606:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 11 25 theme appropriate 1609:1619 arg1 vacuole 1595:1601 the central vacuole 1583:1601 the central vacuole 1583:1601 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 6 26 theme plant 833:837 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 8 27 theme RFP 1106:1108 arg1 fusions 1110:1116 RFP fusions 1106:1116 RFP fusions 1106:1116 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 3 28 dep benthamiana 485:495 arg1 leaves 497:502 leaves 497:502 leaves 497:502 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 3 29 theme vacuolar 395:402 arg1 targeting 410:418 vacuolar (vac) targeting 395:418 vacuolar (vac) targeting 395:418 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 11 30 theme glycan 1785:1790 arg1 processing 1792:1801 plant glycan processing 1779:1801 plant glycan processing 1779:1801 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 11 31 theme appropriate 1674:1684 arg1 modifications 1705:1717 appropriate post-translational modifications 1674:1717 appropriate post-translational modifications 1674:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 0 32 theme recombinant 22:32 arg1 antibodies 34:43 recombinant antibodies 22:43 recombinant antibodies 22:43 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana. 27159528 8 33 located detected 1202:1209 arg1 vacuole 1226:1232 the central vacuole 1214:1232 the central vacuole 1214:1232 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 8 33 located detected 1202:1209 arg2 vac-Abs-RFP 1173:1183 vac-Abs-RFP 1173:1183 vac-Abs-RFP 1173:1183 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 11 34 from reconsideration 1740:1754 arg1 processing 1792:1801 plant glycan processing 1779:1801 plant glycan processing 1779:1801 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 9 35 contain containing 1375:1384 arg2 structures 1390:1399 Man structures 1386:1399 Man structures 1386:1399 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 35 contain containing 1375:1384 arg1 molecules 1365:1373 Ab molecules 1362:1373 Ab molecules containing Man structures 1362:1399 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 2 36 theme processes 315:323 arg1 understanding 284:296 a better understanding 275:296 a better understanding of intracellular processes that affect protein properties 275:354 However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. 27159528 4 37 theme 14D9 608:611 arg1 chain 599:603 the heavy chain 589:603 the heavy chain of 14D9 (vac-Abs) 589:621 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 6 38 from presence 821:828 arg1 sec-Ab 904:909 sec-Ab 904:909 sec-Ab 904:909 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 9 39 theme complex 1465:1471 arg1 N-glycans 1473:1481 complex N-glycans 1465:1481 complex N-glycans 1465:1481 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 40 theme Ab 1441:1442 arg1 molecules 1444:1452 Ab molecules 1441:1452 Ab molecules containing complex N-glycans 1441:1481 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 2 41 theme intracellular 301:313 arg1 processes 315:323 intracellular processes 301:323 intracellular processes that affect protein properties 301:354 However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. 27159528 5 42 theme vac-Abs 730:736 arg1 Accumulation 706:717 Accumulation 706:717 Accumulation of ER- and vac-Abs 706:736 Accumulation of ER- and vac-Abs was 10- to 15-fold higher than sec-Ab. 27159528 11 43 theme plant 1779:1783 arg1 processing 1792:1801 plant glycan processing 1779:1801 plant glycan processing 1779:1801 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 9 44 contain containing 1454:1463 arg1 molecules 1444:1452 Ab molecules 1441:1452 Ab molecules containing complex N-glycans 1441:1481 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 44 contain containing 1454:1463 arg2 N-glycans 1473:1481 complex N-glycans 1465:1481 complex N-glycans 1465:1481 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 4 45 dep C-terminal 569:578 arg1 fused 580:584 fused 580:584 fused to the heavy chain of 14D9 (vac-Abs) 580:621 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 45 dep C-terminal 569:578 arg1 compared 627:634 compared 627:634 compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively) 627:703 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 11 46 with production 1651:1660 arg1 modifications 1705:1717 appropriate post-translational modifications 1674:1717 appropriate post-translational modifications 1674:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 3 47 theme monoclonal 442:451 arg1 antibody 453:460 monoclonal antibody 442:460 the monoclonal antibody (Ab) 14D9 438:470 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 3 47 theme monoclonal 442:451 arg1 Ab 463:464 Ab 463:464 Ab 463:464 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 0 48 theme antibodies 34:43 arg1 targeting 9:17 Vacuolar targeting 0:17 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana 0:68 Vacuolar targeting of recombinant antibodies in Nicotiana benthamiana. 27159528 4 49 theme vacuolar 518:525 arg1 KISIA 546:550 KISIA 546:550 KISIA 546:550 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 49 theme vacuolar 518:525 arg1 signals 537:543 Two distinct vacuolar targeting signals 505:543 Two distinct vacuolar targeting signals (KISIA and NIFRGF) 505:562 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 49 theme vacuolar 518:525 arg1 NIFRGF 556:561 NIFRGF 556:561 NIFRGF 556:561 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 11 50 theme concepts 1767:1774 arg1 reconsideration 1740:1754 a reconsideration 1738:1754 a reconsideration of current concepts in plant glycan processing 1738:1801 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 6 51 theme predominant 809:819 arg1 presence 821:828 the predominant presence 805:828 the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab 805:909 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 9 52 theme Ab 1362:1363 arg1 molecules 1365:1373 Ab molecules 1362:1373 Ab molecules containing Man structures 1362:1399 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 3 53 theme antibody 453:460 arg1 14D9 467:470 the monoclonal antibody (Ab) 14D9 438:470 the monoclonal antibody (Ab) 14D9 438:470 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 4 54 theme distinct 509:516 arg1 KISIA 546:550 KISIA 546:550 KISIA 546:550 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 54 theme distinct 509:516 arg1 signals 537:543 Two distinct vacuolar targeting signals 505:543 Two distinct vacuolar targeting signals (KISIA and NIFRGF) 505:562 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 54 theme distinct 509:516 arg1 NIFRGF 556:561 NIFRGF 556:561 NIFRGF 556:561 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 8 55 theme Confocal 1071:1078 arg1 microscopy 1080:1089 Confocal microscopy 1071:1089 Confocal microscopy analysis using RFP fusions 1071:1116 Confocal microscopy analysis using RFP fusions showed that sec-Ab-RFP localized in the apoplast while vac-Abs-RFP were exclusively detected in the central vacuole. 27159528 9 56 theme direct 1312:1317 arg1 transport 1319:1327 direct transport 1312:1327 direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) 1312:1400 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 56 theme direct 1312:1317 arg1 pathways 1302:1309 two different pathways 1288:1309 two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking 1288:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 57 theme different 1292:1300 arg1 transport 1319:1327 direct transport 1312:1327 direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) 1312:1400 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 57 theme different 1292:1300 arg1 trafficking 1406:1416 trafficking 1406:1416 trafficking 1406:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 9 57 theme different 1292:1300 arg1 pathways 1302:1309 two different pathways 1288:1309 two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking 1288:1416 The data suggest that vac-Abs reached the vacuole by two different pathways: direct transport from the ER bypassing the Golgi (Ab molecules containing Man structures) and trafficking through the Golgi (for Ab molecules containing complex N-glycans). 27159528 3 58 theme vac 405:407 arg1 targeting 410:418 vacuolar (vac) targeting 395:418 vacuolar (vac) targeting 395:418 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 6 59 theme structures 890:899 arg1 presence 821:828 the predominant presence 805:828 the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab 805:909 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 6 60 theme xylosylated 871:881 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 3 61 theme Nicotiana 475:483 arg1 benthamiana 485:495 Nicotiana benthamiana 475:495 Nicotiana benthamiana leaves 475:502 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 4 62 theme targeting 527:535 arg1 KISIA 546:550 KISIA 546:550 KISIA 546:550 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 62 theme targeting 527:535 arg1 signals 537:543 Two distinct vacuolar targeting signals 505:543 Two distinct vacuolar targeting signals (KISIA and NIFRGF) 505:562 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 62 theme targeting 527:535 arg1 NIFRGF 556:561 NIFRGF 556:561 NIFRGF 556:561 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 7 63 theme Paucimannosidic 988:1002 arg1 glycans 1004:1010 Paucimannosidic glycans 988:1010 Paucimannosidic glycans (commonly assigned as typical vacuolar) 988:1050 Paucimannosidic glycans (commonly assigned as typical vacuolar) were not detected. 27159528 1 64 theme recombinant 136:146 arg1 proteins 148:155 recombinant proteins 136:155 recombinant proteins 136:155 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 1 64 theme recombinant 136:146 arg1 antibodies 179:188 monoclonal antibodies 168:188 monoclonal antibodies 168:188 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 6 65 gly fucosylated 855:865 arg1 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 4 66 dep secreted 641:648 arg1 sec-Ab 676:681 sec-Ab 676:681 sec-Ab 676:681 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 66 dep secreted 641:648 arg1 ER-Ab 684:688 ER-Ab 684:688 ER-Ab 684:688 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 6 67 theme N-glycan 777:784 arg1 profiling 786:794 N-glycan profiling 777:794 N-glycan profiling 777:794 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 11 68 theme central 1587:1593 arg1 compartment 1621:1631 an appropriate compartment 1606:1631 an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications 1606:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 11 68 theme central 1587:1593 arg1 vacuole 1595:1601 the central vacuole 1583:1601 the central vacuole 1583:1601 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 2 69 theme protein 337:343 arg1 properties 345:354 protein properties 337:354 protein properties 337:354 However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. 27159528 1 70 used used 109:112 arg2 platforms 83:91 Plant-based platforms 71:91 Plant-based platforms 71:91 Plant-based platforms are extensively used for the expression of recombinant proteins, including monoclonal antibodies. 27159528 7 71 dep assigned 1022:1029 arg1 commonly 1013:1020 commonly 1013:1020 commonly 1013:1020 Paucimannosidic glycans (commonly assigned as typical vacuolar) were not detected. 27159528 6 72 dep oligomannosidic 940:954 arg1 Man 957:959 Man 7-9 957:963 Man 7-9 957:963 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 2 73 theme full 259:262 arg1 potential 264:272 its full potential 255:272 its full potential 255:272 However, to harness the approach effectively and leverage it to its full potential, a better understanding of intracellular processes that affect protein properties is required. 27159528 3 74 from deposition 424:433 arg1 benthamiana 485:495 Nicotiana benthamiana 475:495 Nicotiana benthamiana leaves 475:502 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 6 75 attach presence 821:828 arg2 structures 890:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 plant typical complex fucosylated and xylosylated GnGnXF structures 833:899 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 6 75 attach presence 821:828 arg1 sec-Ab 904:909 sec-Ab 904:909 sec-Ab 904:909 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 3 76 theme 14D9 467:470 arg1 deposition 424:433 deposition 424:433 deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves 424:502 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 3 76 theme 14D9 467:470 arg1 targeting 410:418 vacuolar (vac) targeting 395:418 vacuolar (vac) targeting 395:418 In this work, we examined vacuolar (vac) targeting and deposition of the monoclonal antibody (Ab) 14D9 in Nicotiana benthamiana leaves. 27159528 6 77 theme GnGnXF 974:979 arg1 forms 981:985 GnGnXF forms 974:985 GnGnXF forms 974:985 N-glycan profiling revealed the predominant presence of plant typical complex fucosylated and xylosylated GnGnXF structures on sec-Ab while vac-Abs carried mainly oligomannosidic (Man 7-9) next to GnGnXF forms. 27159528 4 78 dep signals 537:543 arg1 KISIA 546:550 KISIA 546:550 KISIA 546:550 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 78 dep signals 537:543 arg1 signals 537:543 Two distinct vacuolar targeting signals 505:543 Two distinct vacuolar targeting signals (KISIA and NIFRGF) 505:562 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 4 78 dep signals 537:543 arg1 NIFRGF 556:561 NIFRGF 556:561 NIFRGF 556:561 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 27159528 11 79 theme Abs 1665:1667 arg1 production 1651:1660 the efficient production 1637:1660 the efficient production of Abs with appropriate post-translational modifications 1637:1717 Collectively, we show that the central vacuole is an appropriate compartment for the efficient production of Abs with appropriate post-translational modifications, but also point to a reconsideration of current concepts in plant glycan processing. 27159528 4 80 theme heavy 593:597 arg1 chain 599:603 the heavy chain 589:603 the heavy chain of 14D9 (vac-Abs) 589:621 Two distinct vacuolar targeting signals (KISIA and NIFRGF) were C-terminal fused to the heavy chain of 14D9 (vac-Abs) and compared with secreted and ER-retained variants (sec-Ab, ER-Ab, respectively). 25530549 7 0 theme oligosaccharidic 1148:1163 arg1 structures 1165:1174 oligosaccharidic structures 1148:1174 oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1148:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 4 1 theme electrospray 617:628 arg1 source 647:652 an electrospray ionization (ESI) source 614:652 an electrospray ionization (ESI) source 614:652 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 2 2 theme compounds 279:287 arg1 removal 259:265 removal 259:265 removal of phenolic compounds 259:287 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 4 3 theme mass 670:673 arg1 analyzer 675:682 an ion trap mass analyzer 658:682 an ion trap mass analyzer 658:682 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 0 4 theme Complex 0:6 arg1 carbohydrates 8:20 Complex carbohydrates 0:20 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 0:114 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 1 5 theme wine 169:172 arg1 oligosaccharides 134:149 The major neutral oligosaccharides 116:149 The major neutral oligosaccharides of a Carignan red wine 116:172 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 8 6 theme wine 1272:1275 arg1 oligosaccharides 1277:1292 The wine oligosaccharides 1268:1292 The wine oligosaccharides identified 1268:1303 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 7 7 theme oligosaccharide 1103:1117 arg1 fractions 1119:1127 the oligosaccharide fractions 1099:1127 the oligosaccharide fractions 1099:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 2 8 theme polyamide 292:300 arg1 chromatography 302:315 polyamide chromatography 292:315 polyamide chromatography 292:315 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 5 9 theme predominant 879:889 arg1 presence 891:898 the predominant presence 875:898 the predominant presence of arabinose 875:911 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 4 10 theme ion 661:663 arg1 analyzer 675:682 an ion trap mass analyzer 658:682 an ion trap mass analyzer 658:682 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 11 theme ion 1087:1089 arg1 mode 1091:1094 the negative ion mode 1074:1094 the negative ion mode of the oligosaccharide fractions 1074:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 1 12 theme first 206:210 arg1 time 212:215 the first time 202:215 the first time 202:215 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 4 13 theme ionization 630:639 arg1 source 647:652 an electrospray ionization (ESI) source 614:652 an electrospray ionization (ESI) source 614:652 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 1 14 theme neutral 126:132 arg1 oligosaccharides 134:149 The major neutral oligosaccharides 116:149 The major neutral oligosaccharides of a Carignan red wine 116:172 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 1 15 theme major 120:124 arg1 oligosaccharides 134:149 The major neutral oligosaccharides 116:149 The major neutral oligosaccharides of a Carignan red wine 116:172 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 8 16 theme different 1382:1390 arg1 rhamnogalacturonan-arabino-oligosaccharides 1392:1434 different rhamnogalacturonan-arabino-oligosaccharides 1382:1434 different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49 1382:1463 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 4 17 theme interaction 716:726 arg1 chromatography 735:748 hydrophilic interaction liquid chromatography 704:748 hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 704:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 0 18 theme diversity 67:75 arg1 characterization 35:50 characterization 35:50 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 0:114 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 1 19 theme Carignan 156:163 arg1 wine 169:172 a Carignan red wine 154:172 a Carignan red wine 154:172 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 6 20 theme mg/L 1037:1040 arg1 concentration 1016:1028 a concentration 1014:1028 a concentration of 185 mg/L in this wine 1014:1053 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 4 21 theme sulfoalkyl 792:801 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 21 theme sulfoalkyl 792:801 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 0 22 theme neutral 80:86 arg1 oligosaccharides 88:103 neutral oligosaccharides 80:103 neutral oligosaccharides 80:103 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 4 23 theme stationary 811:820 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 23 theme stationary 811:820 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 24 theme unit 1235:1238 arg1 -α-d-GalpA 1256:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 4 25 theme HILIC 765:769 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 25 theme HILIC 765:769 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 2 26 theme anion 395:399 arg1 exchange 401:408 anion exchange 395:408 anion exchange 395:408 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 5 27 theme composition 847:857 arg1 analysis 859:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 5 28 from mannose 944:950 arg1 proportion 961:970 lower proportion 955:970 lower proportion 955:970 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 4 29 theme mass 586:589 arg1 spectrometry 591:602 mass spectrometry 586:602 mass spectrometry (MS) 586:607 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 29 theme mass 586:589 arg1 MS 605:606 MS 605:606 MS 605:606 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 0 30 dep carbohydrates 8:20 arg1 characterization 35:50 characterization 35:50 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 0:114 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 6 31 from concentration 1016:1028 arg1 wine 1050:1053 this wine 1045:1053 this wine 1045:1053 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 6 31 from concentration 1016:1028 arg1 present 1003:1009 present 1003:1009 present 1003:1009 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 8 32 with rhamno-arabino-oligosaccharides 1345:1375 arg1 DP 1441:1442 DP 1441:1442 DP ranging from 5 to 49 1441:1463 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 1 33 theme red 165:167 arg1 wine 169:172 a Carignan red wine 154:172 a Carignan red wine 154:172 The major neutral oligosaccharides of a Carignan red wine have been characterized for the first time. 25530549 0 34 theme wine 29:32 arg1 carbohydrates 8:20 Complex carbohydrates 0:20 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 0:114 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 8 35 with arabino-oligosaccharides 1319:1342 arg1 DP 1441:1442 DP 1441:1442 DP ranging from 5 to 49 1441:1463 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 3 36 theme wine 504:507 arg1 oligosaccharides 509:524 wine oligosaccharides 504:524 wine oligosaccharides 504:524 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 5 37 theme Glycosyl 830:837 arg1 analysis 859:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 8 38 with rhamnogalacturonan-arabino-oligosaccharides 1392:1434 arg1 DP 1441:1442 DP 1441:1442 DP ranging from 5 to 49 1441:1463 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 8 39 theme cell 1506:1509 arg1 pectins 1516:1522 grape cell wall pectins 1500:1522 grape cell wall pectins 1500:1522 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 7 40 from spectra 1063:1069 arg1 mode 1091:1094 the negative ion mode 1074:1094 the negative ion mode of the oligosaccharide fractions 1074:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 2 41 theme size-exclusion 414:427 arg1 chromatography 429:442 size-exclusion chromatography 414:442 size-exclusion chromatography 414:442 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 8 42 theme pectins 1516:1522 arg1 degradation 1485:1495 the degradation 1481:1495 the degradation of grape cell wall pectins 1481:1522 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 4 43 theme ESI 642:644 arg1 source 647:652 an electrospray ionization (ESI) source 614:652 an electrospray ionization (ESI) source 614:652 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 44 theme fractions 1119:1127 arg1 mode 1091:1094 the negative ion mode 1074:1094 the negative ion mode of the oligosaccharide fractions 1074:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 2 45 theme phenolic 270:277 arg1 compounds 279:287 phenolic compounds 270:287 phenolic compounds 270:287 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 5 46 theme arabinose 903:911 arg1 presence 891:898 the predominant presence 875:898 the predominant presence of arabinose 875:911 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 4 47 theme trap 665:668 arg1 analyzer 675:682 an ion trap mass analyzer 658:682 an ion trap mass analyzer 658:682 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 48 theme → 1252:1252 arg1 -α-d-GalpA 1256:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 8 49 dep 49 1462:1463 arg1 to 1459:1460 to 1459:1460 to 1459:1460 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 9 50 theme extreme 1550:1556 arg1 diversity 1558:1566 an extreme diversity 1547:1566 an extreme diversity 1547:1566 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 2 51 theme alcohol 343:349 arg1 precipitation 351:363 alcohol precipitation 343:363 alcohol precipitation 343:363 The oligosaccharides were prepared after removal of phenolic compounds by polyamide chromatography and of polysaccharides by alcohol precipitation and then were fractionated by anion exchange and size-exclusion chromatography. 25530549 7 52 theme negative 1078:1085 arg1 mode 1091:1094 the negative ion mode 1074:1094 the negative ion mode of the oligosaccharide fractions 1074:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 7 53 theme α-l-Rhap- 1240:1248 arg1 -α-d-GalpA 1256:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 6 54 theme Neutral 973:979 arg1 oligosaccharides 981:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 9 55 located detected 1594:1601 arg2 peaks 1588:1592 more than 100 peaks 1574:1592 more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure 1574:1686 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 9 55 located detected 1594:1601 arg1 spectra 1618:1624 HPLC-ESI-MS spectra 1606:1624 HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure 1606:1686 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 4 56 theme liquid 728:733 arg1 chromatography 735:748 hydrophilic interaction liquid chromatography 704:748 hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 704:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 57 theme MS 1060:1061 arg1 spectra 1063:1069 The MS spectra 1056:1069 The MS spectra in the negative ion mode of the oligosaccharide fractions 1056:1127 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 5 58 theme lower 955:959 arg1 proportion 961:970 lower proportion 955:970 lower proportion 955:970 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 0 59 theme extreme 59:65 arg1 diversity 67:75 the extreme diversity 55:75 the extreme diversity of neutral oligosaccharides 55:103 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 6 60 attach present 1003:1009 arg1 concentration 1016:1028 a concentration 1014:1028 a concentration of 185 mg/L in this wine 1014:1053 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 6 60 attach present 1003:1009 arg2 oligosaccharides 981:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 5 61 from galactose 919:927 arg1 proportion 961:970 lower proportion 955:970 lower proportion 955:970 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 9 62 theme HPLC-ESI-MS 1606:1616 arg1 spectra 1618:1624 HPLC-ESI-MS spectra 1606:1624 HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure 1606:1686 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 4 63 theme Oligosaccharide 543:557 arg1 fractions 559:567 Oligosaccharide fractions 543:567 Oligosaccharide fractions 543:567 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 9 64 contain have 1542:1545 arg2 diversity 1558:1566 an extreme diversity 1547:1566 an extreme diversity 1547:1566 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 9 64 contain have 1542:1545 arg1 Oligosaccharides 1525:1540 Oligosaccharides 1525:1540 Oligosaccharides 1525:1540 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 4 65 theme hydrophilic 704:714 arg1 chromatography 735:748 hydrophilic interaction liquid chromatography 704:748 hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 704:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 0 66 theme oligosaccharides 88:103 arg1 diversity 67:75 the extreme diversity 55:75 the extreme diversity of neutral oligosaccharides 55:103 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 4 67 theme zwitterionic 779:790 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 67 theme zwitterionic 779:790 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 7 68 theme basic 1229:1233 arg1 -α-d-GalpA 1256:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 5 69 from rhamnose 930:937 arg1 proportion 961:970 lower proportion 955:970 lower proportion 955:970 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 4 70 theme betaine 803:809 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 70 theme betaine 803:809 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 5 71 theme residue 839:845 arg1 analysis 859:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis 830:866 Glycosyl residue composition analysis showed the predominant presence of arabinose, with galactose, rhamnose, and mannose in lower proportion. 25530549 7 72 dep -α-d-GalpA 1256:1265 arg1 4 1254:1254 4 1254:1254 4 1254:1254 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 7 73 attach linked 1215:1220 arg1 -α-d-GalpA 1256:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1225:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 7 73 attach linked 1215:1220 arg2 oligo-arabinans 1193:1207 oligo-arabinans 1193:1207 oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1193:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 4 74 theme Nucleodur 755:763 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 74 theme Nucleodur 755:763 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 3 75 theme second 450:455 arg1 step 457:460 a second step 448:460 a second step 448:460 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 0 76 theme red 25:27 arg1 wine 29:32 red wine 25:32 red wine 25:32 Complex carbohydrates of red wine: characterization of the extreme diversity of neutral oligosaccharides by ESI-MS. 25530549 6 77 located present 1003:1009 arg2 oligosaccharides 981:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides 973:996 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 6 77 located present 1003:1009 arg1 concentration 1016:1028 a concentration 1014:1028 a concentration of 185 mg/L in this wine 1014:1053 Neutral oligosaccharides were present at a concentration of 185 mg/L in this wine. 25530549 9 78 theme oligosaccharidic 1661:1676 arg1 structure 1678:1686 at least one oligosaccharidic structure 1648:1686 at least one oligosaccharidic structure 1648:1686 Oligosaccharides have an extreme diversity, with more than 100 peaks detected in HPLC-ESI-MS spectra corresponding each to at least one oligosaccharidic structure. 25530549 3 79 theme glycosyl 467:474 arg1 composition 476:486 glycosyl composition 467:486 glycosyl composition 467:486 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 7 80 theme structures 1165:1174 arg1 series 1138:1143 a series 1136:1143 a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA 1136:1265 The MS spectra in the negative ion mode of the oligosaccharide fractions showed a series of oligosaccharidic structures corresponding to oligo-arabinans often linked to the basic unit α-l-Rhap-(1 → 4)-α-d-GalpA. 25530549 3 81 theme oligosaccharides 509:524 arg1 linkages 492:499 linkages 492:499 linkages 492:499 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 3 81 theme oligosaccharides 509:524 arg1 composition 476:486 glycosyl composition 467:486 glycosyl composition 467:486 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 8 82 theme grape 1500:1504 arg1 pectins 1516:1522 grape cell wall pectins 1500:1522 grape cell wall pectins 1500:1522 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 25530549 4 83 from chromatography 735:748 arg1 column 771:776 a Nucleodur HILIC column 753:776 a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase) 753:827 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 4 83 from chromatography 735:748 arg1 phase 822:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 zwitterionic sulfoalkyl betaine stationary phase 779:826 Oligosaccharide fractions were analyzed by mass spectrometry (MS) with an electrospray ionization (ESI) source and an ion trap mass analyzer after separation by hydrophilic interaction liquid chromatography on a Nucleodur HILIC column (zwitterionic sulfoalkyl betaine stationary phase). 25530549 3 84 dep composition 476:486 arg1 the 463:465 the 463:465 the 463:465 In a second step, the glycosyl composition and linkages of wine oligosaccharides were determined. 25530549 8 85 theme wall 1511:1514 arg1 pectins 1516:1522 grape cell wall pectins 1500:1522 grape cell wall pectins 1500:1522 The wine oligosaccharides identified correspond to arabino-oligosaccharides, rhamno-arabino-oligosaccharides, and different rhamnogalacturonan-arabino-oligosaccharides with DP ranging from 5 to 49, resulting from the degradation of grape cell wall pectins. 28849661 1 0 theme chondroitin 79:89 arg1 glycopeptide 99:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 1 1 theme first 136:140 arg1 time 142:145 the first time 132:145 the first time using the cassette approach 132:173 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 2 2 theme glycosylation 192:204 arg1 critical 253:260 critical 253:260 critical 253:260 The sequence of glycosylation to form the octasaccharide serine cassette was critical. 28849661 2 2 theme glycosylation 192:204 arg1 sequence 180:187 The sequence 176:187 The sequence of glycosylation to form the octasaccharide serine cassette 176:247 The sequence of glycosylation to form the octasaccharide serine cassette was critical. 28849661 3 3 theme 2+ 313:314 arg1 strategy 338:345 a 2+ (3 + 3) glycosylation strategy 311:345 a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation 311:382 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 0 4 theme Bearing 35:41 arg1 Glycopeptide 54:65 A Bearing Syndecan-1 Glycopeptide 33:65 A Bearing Syndecan-1 Glycopeptide 33:65 Synthesis of Chondroitin Sulfate A Bearing Syndecan-1 Glycopeptide. 28849661 1 5 theme sulfate 91:97 arg1 glycopeptide 99:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 3 6 theme chain 367:371 arg1 elongation 373:382 peptide chain elongation 359:382 peptide chain elongation 359:382 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 0 7 dep Glycopeptide 54:65 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Chondroitin Sulfate 0:31 Synthesis of Chondroitin Sulfate A Bearing Syndecan-1 Glycopeptide. 28849661 2 8 theme serine 233:238 arg1 cassette 240:247 the octasaccharide serine cassette 214:247 the octasaccharide serine cassette 214:247 The sequence of glycosylation to form the octasaccharide serine cassette was critical. 28849661 3 9 theme peptide 359:365 arg1 elongation 373:382 peptide chain elongation 359:382 peptide chain elongation 359:382 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 3 10 dep strategy 338:345 arg1 3 321:321 3 321:321 3 321:321 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 3 10 dep strategy 338:345 arg1 3 317:317 3 317:317 3 317:317 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 3 11 gly glycopeptide 267:278 arg2 glycopeptide 267:278 The glycopeptide 263:278 The glycopeptide 263:278 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 3 12 theme glycosylation 324:336 arg1 strategy 338:345 a 2+ (3 + 3) glycosylation strategy 311:345 a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation 311:382 The glycopeptide was successfully assembled via a 2+ (3 + 3) glycosylation strategy followed by peptide chain elongation. 28849661 1 13 gly glycopeptide 99:110 arg2 glycopeptide 99:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 0 14 theme Sulfate 25:31 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Chondroitin Sulfate 0:31 Synthesis of Chondroitin Sulfate A Bearing Syndecan-1 Glycopeptide. 28849661 1 15 theme Syndecan-1 68:77 arg1 glycopeptide 99:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide 68:110 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 0 16 theme Syndecan-1 43:52 arg1 Glycopeptide 54:65 A Bearing Syndecan-1 Glycopeptide 33:65 A Bearing Syndecan-1 Glycopeptide 33:65 Synthesis of Chondroitin Sulfate A Bearing Syndecan-1 Glycopeptide. 28849661 1 17 theme cassette 157:164 arg1 approach 166:173 the cassette approach 153:173 the cassette approach 153:173 Syndecan-1 chondroitin sulfate glycopeptide was synthesized for the first time using the cassette approach. 28849661 2 18 theme octasaccharide 218:231 arg1 cassette 240:247 the octasaccharide serine cassette 214:247 the octasaccharide serine cassette 214:247 The sequence of glycosylation to form the octasaccharide serine cassette was critical. 28849661 0 19 theme Chondroitin 13:23 arg1 Sulfate 25:31 Chondroitin Sulfate 13:31 Chondroitin Sulfate 13:31 Synthesis of Chondroitin Sulfate A Bearing Syndecan-1 Glycopeptide. 26747427 0 0 theme nonfucosylated 93:106 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 6 1 theme higher 982:987 arg1 affinity 1005:1012 higher heparin-binding affinity 982:1012 higher heparin-binding affinity 982:1012 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 4 2 theme AT 754:755 arg1 activities 740:749 biological activities 729:749 biological activities of AT 729:755 However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. 26747427 0 3 link N-linked 108:115 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 3 4 theme α-form 524:529 arg1 mixture 509:515 a mixture 507:515 a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135 507:611 In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. 26747427 6 5 theme high-mannose 957:968 arg1 type 970:973 the high-mannose type 953:973 the high-mannose type 953:973 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 0 6 theme complex-type 80:91 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 9 7 theme plasma 1383:1388 arg1 half-life 1390:1398 a prolonged plasma half-life 1371:1398 a prolonged plasma half-life 1371:1398 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 26747427 10 8 theme AT 1573:1574 arg1 activities 1590:1599 AT physiological activities 1573:1599 AT physiological activities 1573:1599 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 8 9 theme high-mannose 1262:1273 arg1 type 1275:1278 the high-mannose type 1258:1278 the high-mannose type 1258:1278 In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. 26747427 0 10 theme N-linked 108:115 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 1 11 attach attached 182:189 arg1 AT 208:209 AT 208:209 AT 208:209 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 1 11 attach attached 182:189 arg2 oligosaccharides 165:180 the N-linked oligosaccharides 152:180 the N-linked oligosaccharides attached to antithrombin (AT) 152:210 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 1 11 attach attached 182:189 arg1 antithrombin 194:205 antithrombin 194:205 antithrombin (AT) 194:210 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 7 12 theme high-mannose 1209:1220 arg1 type 1222:1225 the high-mannose type 1205:1225 the high-mannose type 1205:1225 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 10 13 theme high-mannose-type 1687:1703 arg1 structures 1705:1714 the high-mannose-type structures 1683:1714 the high-mannose-type structures at the nonreducing end 1683:1737 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 2 14 theme complex-type 311:322 arg1 oligosaccharides 324:339 biantennary complex-type oligosaccharides 299:339 biantennary complex-type oligosaccharides 299:339 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 4 15 theme immature 650:657 arg1 oligosaccharides 677:692 the immature high-mannose-type oligosaccharides 646:692 the immature high-mannose-type oligosaccharides produced by mammalian cells 646:720 However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. 26747427 2 16 theme biantennary 299:309 arg1 oligosaccharides 324:339 biantennary complex-type oligosaccharides 299:339 biantennary complex-type oligosaccharides 299:339 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 10 17 theme nonreducing 1723:1733 arg1 end 1735:1737 the nonreducing end 1719:1737 the nonreducing end 1719:1737 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 3 18 theme human 476:480 arg1 plasma 482:487 human plasma 476:487 human plasma 476:487 In human plasma, AT circulates as a mixture of the α-form bearing four oligosaccharides and the β-form lacking an oligosaccharide at Asn135. 26747427 5 19 theme high-mannose 826:837 arg1 types 851:855 the high-mannose and complex types 822:855 types 851:855 Here, we succeeded in directly comparing the activities between the high-mannose and complex types. 26747427 0 20 gly nonfucosylated 93:106 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 10 21 theme core 1639:1642 arg1 fucose 1644:1649 a core fucose 1637:1649 a core fucose at the reducing end 1637:1669 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 10 22 theme present 1543:1549 arg1 study 1551:1555 The present study 1539:1555 The present study 1539:1555 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 7 23 theme anticoagulant 1019:1031 arg1 activities 1033:1042 The anticoagulant activities 1015:1042 The anticoagulant activities 1015:1042 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 7 24 theme strongest 1137:1145 arg1 activity 1161:1168 the strongest anticoagulant activity 1133:1168 the strongest anticoagulant activity 1133:1168 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 2 25 theme biological 409:418 arg1 activities 420:429 its biological activities 405:429 its biological activities 405:429 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 6 26 from differences 908:918 arg1 activity 943:950 thrombin inhibitory activity 923:950 thrombin inhibitory activity 923:950 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 2 27 contain has 295:297 arg2 oligosaccharides 324:339 biantennary complex-type oligosaccharides 299:339 biantennary complex-type oligosaccharides 299:339 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 2 27 contain has 295:297 arg1 AT 292:293 Human AT 286:293 Human AT 286:293 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 2 28 theme unique 350:355 arg1 feature 357:363 the unique feature 346:363 the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity 346:470 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 0 29 theme activities 25:34 arg1 Comparison 0:9 Comparison 0:9 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 0:133 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 4 30 theme high-mannose-type 659:675 arg1 oligosaccharides 677:692 the immature high-mannose-type oligosaccharides 646:692 the immature high-mannose-type oligosaccharides produced by mammalian cells 646:720 However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. 26747427 0 31 theme biological 14:23 arg1 activities 25:34 biological activities 14:34 biological activities of human antithrombins 14:57 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 9 32 theme high-mannose 1433:1444 arg1 type 1446:1449 the high-mannose type 1429:1449 the high-mannose type 1429:1449 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 26747427 10 33 theme physiological 1576:1588 arg1 activities 1590:1599 AT physiological activities 1573:1599 AT physiological activities 1573:1599 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 6 34 theme inhibitory 932:941 arg1 activity 943:950 thrombin inhibitory activity 923:950 thrombin inhibitory activity 923:950 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 2 35 theme heparin-binding 447:461 arg1 affinity 463:470 its heparin-binding affinity 443:470 its heparin-binding affinity 443:470 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 0 36 theme human 39:43 arg1 antithrombins 45:57 human antithrombins 39:57 human antithrombins 39:57 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 11 37 theme human 1869:1873 arg1 plasma 1875:1880 human plasma 1869:1880 human plasma 1869:1880 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 7 38 theme anticoagulant 1147:1159 arg1 activity 1161:1168 the strongest anticoagulant activity 1133:1168 the strongest anticoagulant activity 1133:1168 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 6 39 theme thrombin 923:930 arg1 activity 943:950 thrombin inhibitory activity 923:950 thrombin inhibitory activity 923:950 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 4 40 theme mammalian 706:714 arg1 cells 716:720 mammalian cells 706:720 mammalian cells 706:720 However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. 26747427 11 41 theme immature 1760:1767 arg1 type 1782:1785 the immature high-mannose type 1756:1785 the immature high-mannose type 1756:1785 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 8 42 theme plasma 1302:1307 arg1 half-life 1309:1317 a much shorter plasma half-life 1287:1317 a much shorter plasma half-life than the complex type 1287:1339 In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. 26747427 11 43 located found 1860:1864 arg1 plasma 1875:1880 human plasma 1869:1880 human plasma 1869:1880 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 11 43 located found 1860:1864 arg2 AT 1847:1848 the AT 1843:1848 the AT typically found in human plasma 1843:1880 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 5 44 theme complex 843:849 arg1 types 851:855 the high-mannose and complex types 822:855 types 851:855 Here, we succeeded in directly comparing the activities between the high-mannose and complex types. 26747427 10 45 theme reducing 1658:1665 arg1 end 1667:1669 the reducing end 1654:1669 the reducing end 1654:1669 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 11 46 theme high-mannose 1769:1780 arg1 type 1782:1785 the immature high-mannose type 1756:1785 the immature high-mannose type 1756:1785 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 2 47 theme core 378:381 arg1 fucose 383:388 a core fucose 376:388 a core fucose 376:388 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 0 48 theme antithrombins 45:57 arg1 activities 25:34 biological activities 14:34 biological activities of human antithrombins 14:57 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 1 49 link N-linked 156:163 arg1 oligosaccharides 165:180 the N-linked oligosaccharides 152:180 the N-linked oligosaccharides attached to antithrombin (AT) 152:210 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 6 50 theme substantial 896:906 arg1 differences 908:918 no substantial differences 893:918 no substantial differences in thrombin inhibitory activity 893:950 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 9 51 theme prolonged 1373:1381 arg1 half-life 1390:1398 a prolonged plasma half-life 1371:1398 a prolonged plasma half-life 1371:1398 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 26747427 10 52 from end 1667:1669 arg1 fucose 1644:1649 a core fucose 1637:1649 a core fucose at the reducing end 1637:1669 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 0 53 with Comparison 0:9 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 7 54 with β-form 1193:1198 arg1 type 1222:1225 the high-mannose type 1205:1225 the high-mannose type 1205:1225 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 11 55 with β-form 1744:1749 arg1 type 1782:1785 the immature high-mannose type 1756:1785 the immature high-mannose type 1756:1785 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 7 56 theme heparin-binding 1094:1108 arg1 affinity 1110:1117 the heparin-binding affinity 1090:1117 the heparin-binding affinity 1090:1117 The anticoagulant activities were increased by heparin and correlated with the heparin-binding affinity, resulting in the strongest anticoagulant activity being displayed in the β-form with the high-mannose type. 26747427 2 57 theme Human 286:290 arg1 AT 292:293 Human AT 286:293 Human AT 286:293 Human AT has biantennary complex-type oligosaccharides with the unique feature of lacking a core fucose, which affects its biological activities by changing its heparin-binding affinity. 26747427 1 58 theme N-linked 156:163 arg1 oligosaccharides 165:180 the N-linked oligosaccharides 152:180 the N-linked oligosaccharides attached to antithrombin (AT) 152:210 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 8 59 theme complex 1328:1334 arg1 type 1336:1339 the complex type 1324:1339 the complex type 1324:1339 In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. 26747427 1 60 theme anticoagulant 241:253 arg1 activity 255:262 its anticoagulant activity 237:262 its anticoagulant activity 237:262 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 8 61 theme pharmacokinetic 1231:1245 arg1 profiling 1247:1255 pharmacokinetic profiling 1231:1255 pharmacokinetic profiling 1231:1255 In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. 26747427 11 62 theme potent 1817:1822 arg1 anticoagulant 1824:1836 a more potent anticoagulant 1810:1836 a more potent anticoagulant than the AT typically found in human plasma 1810:1880 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 11 62 theme potent 1817:1822 arg1 β-form 1744:1749 The β-form 1740:1749 The β-form with the immature high-mannose type 1740:1785 The β-form with the immature high-mannose type appears to function as a more potent anticoagulant than the AT typically found in human plasma, once it emerges in the blood. 26747427 1 63 theme oligosaccharides 165:180 arg1 structure 139:147 The structure 135:147 The structure of the N-linked oligosaccharides attached to antithrombin (AT) 135:210 The structure of the N-linked oligosaccharides attached to antithrombin (AT) has been shown to affect its anticoagulant activity and pharmacokinetics. 26747427 10 64 from end 1735:1737 arg1 structures 1705:1714 the high-mannose-type structures 1683:1714 the high-mannose-type structures at the nonreducing end 1683:1737 The present study highlights that AT physiological activities are strictly controlled not only by a core fucose at the reducing end but also by the high-mannose-type structures at the nonreducing end. 26747427 0 65 theme high-mannose 64:75 arg1 oligosaccharides 117:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 high-mannose or complex-type nonfucosylated N-linked oligosaccharides 64:132 Comparison of biological activities of human antithrombins with high-mannose or complex-type nonfucosylated N-linked oligosaccharides. 26747427 6 66 theme heparin-binding 989:1003 arg1 affinity 1005:1012 higher heparin-binding affinity 982:1012 higher heparin-binding affinity 982:1012 Interestingly, although there were no substantial differences in thrombin inhibitory activity, the high-mannose type showed higher heparin-binding affinity. 26747427 8 67 theme shorter 1294:1300 arg1 half-life 1309:1317 a much shorter plasma half-life 1287:1317 a much shorter plasma half-life than the complex type 1287:1339 In pharmacokinetic profiling, the high-mannose type showed a much shorter plasma half-life than the complex type. 26747427 4 68 theme biological 729:738 arg1 activities 740:749 biological activities 729:749 biological activities of AT 729:755 However, it remains unclear how the immature high-mannose-type oligosaccharides produced by mammalian cells affect biological activities of AT. 26747427 9 69 contain have 1366:1369 arg1 β-form 1346:1351 The β-form 1342:1351 The β-form 1342:1351 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 26747427 9 69 contain have 1366:1369 arg2 half-life 1390:1398 a prolonged plasma half-life 1371:1398 a prolonged plasma half-life 1371:1398 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 26747427 9 70 theme longer 1484:1489 arg1 half-life 1491:1499 a longer half-life 1482:1499 a longer half-life than the β-form for the complex-type 1482:1536 The β-form was found to have a prolonged plasma half-life compared with the α-form for the high-mannose type; conversely, the α-form showed a longer half-life than the β-form for the complex-type. 25664729 3 0 theme marine 370:375 arg1 member 377:382 This marine member 365:382 This marine member of the Bacteroidetes 365:403 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 10 1 theme initial 1391:1397 arg1 step 1399:1402 the initial step 1387:1402 the initial step of degradation of branched laminarin 1387:1439 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 3 2 dep β-1,3-glucanases 429:444 arg1 β-1,3-glucanases 429:444 five putative β-1,3-glucanases 415:444 five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures 415:551 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 2 dep β-1,3-glucanases 429:444 arg1 GH64 512:515 GH64 512:515 GH64 512:515 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 2 dep β-1,3-glucanases 429:444 arg1 [four 446:450 [four 446:450 [four 446:450 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 9 3 theme moiety 1228:1233 arg1 group 1206:1210 the O6 hydroxyl group 1190:1210 the O6 hydroxyl group of each glucose moiety 1190:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 3 theme moiety 1228:1233 arg1 moiety 1228:1233 each glucose moiety 1215:1233 each glucose moiety 1215:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 7 4 theme mutant 866:871 arg1 structure 841:849 The structure 837:849 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate 837:935 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 1 5 theme occasional 200:209 arg1 branches 217:224 occasional β-1,6 branches 200:224 occasional β-1,6 branches 200:224 Laminarin is a β-1,3-D-glucan displaying occasional β-1,6 branches. 25664729 7 6 theme thio-β-1,3-hexaglucan 905:925 arg1 substrate 927:935 a thio-β-1,3-hexaglucan substrate 903:935 a thio-β-1,3-hexaglucan substrate 903:935 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 8 7 theme a 1078:1078 arg1 glycerol 1080:1087 a glycerol 1078:1087 a glycerol 1078:1087 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 8 7 theme a 1078:1078 arg1 ion 1120:1122 a chloride ion 1109:1122 a chloride ion 1109:1122 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 8 7 theme a 1078:1078 arg1 ion 1101:1103 an acetate ion 1090:1103 an acetate ion 1090:1103 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 8 8 theme lateral 1046:1052 arg1 pockets 1054:1060 These lateral pockets 1040:1060 These lateral pockets 1040:1060 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 9 9 theme hydroxyl 1197:1204 arg1 group 1206:1210 the O6 hydroxyl group 1190:1210 the O6 hydroxyl group of each glucose moiety 1190:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 9 theme hydroxyl 1197:1204 arg1 moiety 1228:1233 each glucose moiety 1215:1233 each glucose moiety 1215:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 0 10 theme preferred 114:122 arg1 recognition 124:134 preferred recognition 114:134 preferred recognition of branched laminarin 114:156 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 9 11 theme group 1206:1210 arg1 vicinity 1178:1185 the vicinity 1174:1185 the vicinity of the O6 hydroxyl group of each glucose moiety 1174:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 10 12 theme secreted 1332:1339 arg1 ZgLamC 1320:1325 ZgLamC 1320:1325 ZgLamC 1320:1325 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 10 12 theme secreted 1332:1339 arg1 laminarinase 1341:1352 a secreted laminarinase 1330:1352 a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins 1330:1545 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 10 12 theme secreted 1332:1339 arg1 likely 1362:1367 likely 1362:1367 likely 1362:1367 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 0 13 theme branched 139:146 arg1 laminarin 148:156 branched laminarin 139:156 branched laminarin 139:156 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 8 14 theme acetate 1093:1099 arg1 glycerol 1080:1087 a glycerol 1078:1087 a glycerol 1078:1087 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 8 14 theme acetate 1093:1099 arg1 ion 1101:1103 an acetate ion 1090:1103 an acetate ion 1090:1103 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 1 15 theme β-1,6 211:215 arg1 branches 217:224 occasional β-1,6 branches 200:224 occasional β-1,6 branches 200:224 Laminarin is a β-1,3-D-glucan displaying occasional β-1,6 branches. 25664729 0 16 theme laminarinase 51:62 arg1 ZgLamCGH16 64:73 the laminarinase ZgLamCGH16 47:73 the laminarinase ZgLamCGH16 from Zobellia galactanivorans 47:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 9 17 attach presence 1143:1150 arg1 vicinity 1178:1185 the vicinity 1174:1185 the vicinity of the O6 hydroxyl group of each glucose moiety 1174:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 17 attach presence 1143:1150 arg2 molecules 1161:1169 these molecules 1155:1169 these molecules 1155:1169 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 7 18 from structure 841:849 arg1 complex 890:896 complex 890:896 complex with a thio-β-1,3-hexaglucan substrate 890:935 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 2 19 theme marine 315:320 arg1 galactanivorans 348:362 Zobellia galactanivorans 339:362 Zobellia galactanivorans 339:362 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 2 19 theme marine 315:320 arg1 bacteria 322:329 marine bacteria 315:329 marine bacteria such as Zobellia galactanivorans 315:362 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 3 20 theme Bacteroidetes 391:403 arg1 member 377:382 This marine member 365:382 This marine member of the Bacteroidetes 365:403 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 6 21 contain has 730:732 arg2 specificity 749:759 a preferential specificity 734:759 a preferential specificity for laminarin but also a significant activity 734:805 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 6 21 contain has 730:732 arg1 enzyme 723:728 This recombinant enzyme 706:728 This recombinant enzyme 706:728 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 5 22 theme catalytic 624:632 arg1 ZgLamCGH16 647:656 ZgLamCGH16 647:656 ZgLamCGH16 647:656 The catalytic GH16 module (ZgLamCGH16) was produced in Escherichia coli and purified. 25664729 5 22 theme catalytic 624:632 arg1 module 639:644 The catalytic GH16 module 620:644 The catalytic GH16 module (ZgLamCGH16) 620:657 The catalytic GH16 module (ZgLamCGH16) was produced in Escherichia coli and purified. 25664729 10 23 theme laminarin 1431:1439 arg1 degradation 1407:1417 degradation 1407:1417 degradation of branched laminarin 1407:1439 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 6 24 theme preferential 736:747 arg1 specificity 749:759 a preferential specificity 734:759 a preferential specificity for laminarin but also a significant activity 734:805 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 0 25 theme Structural 0:9 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans 0:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 5 26 theme GH16 634:637 arg1 ZgLamCGH16 647:656 ZgLamCGH16 647:656 ZgLamCGH16 647:656 The catalytic GH16 module (ZgLamCGH16) was produced in Escherichia coli and purified. 25664729 5 26 theme GH16 634:637 arg1 module 639:644 The catalytic GH16 module 620:644 The catalytic GH16 module (ZgLamCGH16) 620:657 The catalytic GH16 module (ZgLamCGH16) was produced in Escherichia coli and purified. 25664729 10 27 theme characterized 1463:1475 arg1 ZgLamA 1477:1482 the previously characterized ZgLamA 1448:1482 the previously characterized ZgLamA 1448:1482 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 4 28 theme ZgLamC 600:605 arg1 characterization 564:579 the characterization 560:579 the characterization of the β-glucanase ZgLamC 560:605 Here, the characterization of the β-glucanase ZgLamC is reported. 25664729 8 29 theme chloride 1111:1118 arg1 ion 1120:1122 a chloride ion 1109:1122 a chloride ion 1109:1122 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 8 29 theme chloride 1111:1118 arg1 glycerol 1080:1087 a glycerol 1078:1087 a glycerol 1078:1087 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 7 30 theme additional 988:997 arg1 pockets 999:1005 three additional pockets 982:1005 three additional pockets flanking subsites -1, -2 and -3 982:1037 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 7 31 theme ZgLamCGH16 876:885 arg1 mutant 866:871 an inactive mutant 854:871 an inactive mutant of ZgLamCGH16 854:885 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 0 32 theme biochemical 15:25 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans 0:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 0 33 theme laminarin 148:156 arg1 recognition 124:134 preferred recognition 114:134 preferred recognition of branched laminarin 114:156 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 2 34 theme Zobellia 339:346 arg1 galactanivorans 348:362 Zobellia galactanivorans 339:362 Zobellia galactanivorans 339:362 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 7 35 theme inactive 857:864 arg1 mutant 866:871 an inactive mutant 854:871 an inactive mutant of ZgLamCGH16 854:885 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 0 36 from galactanivorans 89:103 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans 0:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 0 36 from galactanivorans 89:103 arg1 ZgLamCGH16 64:73 the laminarinase ZgLamCGH16 47:73 the laminarinase ZgLamCGH16 from Zobellia galactanivorans 47:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 2 37 theme abundant 285:292 arg1 source 294:299 an abundant source 282:299 an abundant source of carbon for marine bacteria such as Zobellia galactanivorans 282:362 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 6 38 theme recombinant 711:721 arg1 enzyme 723:728 This recombinant enzyme 706:728 This recombinant enzyme 706:728 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 9 39 theme O6 1194:1195 arg1 group 1206:1210 the O6 hydroxyl group 1190:1210 the O6 hydroxyl group of each glucose moiety 1190:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 39 theme O6 1194:1195 arg1 moiety 1228:1233 each glucose moiety 1215:1233 each glucose moiety 1215:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 40 from presence 1143:1150 arg1 vicinity 1178:1185 the vicinity 1174:1185 the vicinity of the O6 hydroxyl group of each glucose moiety 1174:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 3 41 contain possesses 405:413 arg2 β-1,3-glucanases 429:444 five putative β-1,3-glucanases 415:444 five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures 415:551 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 41 contain possesses 405:413 arg2 GH64 512:515 GH64 512:515 GH64 512:515 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 41 contain possesses 405:413 arg1 member 377:382 This marine member 365:382 This marine member of the Bacteroidetes 365:403 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 41 contain possesses 405:413 arg2 [four 446:450 [four 446:450 [four 446:450 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 10 42 theme unbranched 1505:1514 arg1 laminarin 1516:1524 unbranched laminarin 1505:1524 unbranched laminarin 1505:1524 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 0 43 theme ZgLamCGH16 64:73 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans 0:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 9 44 theme branched 1273:1280 arg1 substrates 1296:1305 substrates 1296:1305 substrates 1296:1305 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 9 44 theme branched 1273:1280 arg1 laminarins 1282:1291 branched laminarins 1273:1291 branched laminarins 1273:1291 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 6 45 theme mixed-linked 810:821 arg1 MLG 831:833 MLG 831:833 MLG 831:833 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 6 45 theme mixed-linked 810:821 arg1 glucan 823:828 mixed-linked glucan 810:828 mixed-linked glucan (MLG) 810:834 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 3 46 theme modular 531:537 arg1 architectures 539:551 various modular architectures 523:551 various modular architectures 523:551 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 47 theme hydrolase 474:482 arg1 family 484:489 glycosyl hydrolase family 16 465:492 glycosyl hydrolase family 16 (GH16) 465:499 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 47 theme hydrolase 474:482 arg1 GH16 495:498 GH16 495:498 GH16 495:498 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 4 48 theme β-glucanase 588:598 arg1 ZgLamC 600:605 the β-glucanase ZgLamC 584:605 the β-glucanase ZgLamC 584:605 Here, the characterization of the β-glucanase ZgLamC is reported. 25664729 2 49 theme brown 258:262 arg1 algae 264:268 brown algae 258:268 brown algae 258:268 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 2 50 theme algae 264:268 arg1 polysaccharide 240:253 This storage polysaccharide 227:253 This storage polysaccharide of brown algae 227:268 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 9 51 theme molecules 1161:1169 arg1 presence 1143:1150 The presence 1139:1150 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety 1139:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 10 52 theme branched 1422:1429 arg1 laminarin 1431:1439 branched laminarin 1422:1439 branched laminarin 1422:1439 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 7 53 with complex 890:896 arg1 substrate 927:935 a thio-β-1,3-hexaglucan substrate 903:935 a thio-β-1,3-hexaglucan substrate 903:935 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 10 54 theme degradation 1407:1417 arg1 step 1399:1402 the initial step 1387:1402 the initial step of degradation of branched laminarin 1387:1439 Altogether, ZgLamC is a secreted laminarinase that is likely to be involved in the initial step of degradation of branched laminarin, while the previously characterized ZgLamA efficiently degrades unbranched laminarin and oligo-laminarins. 25664729 2 55 theme carbon 304:309 arg1 source 294:299 an abundant source 282:299 an abundant source of carbon for marine bacteria such as Zobellia galactanivorans 282:362 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 6 56 theme significant 786:796 arg1 activity 798:805 a significant activity 784:805 laminarin but also a significant activity 765:805 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 3 57 theme various 523:529 arg1 architectures 539:551 various modular architectures 523:551 various modular architectures 523:551 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 7 58 theme straight 950:957 arg1 cleft 971:975 a straight active-site cleft 948:975 a straight active-site cleft 948:975 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 0 59 theme Zobellia 80:87 arg1 galactanivorans 89:103 Zobellia galactanivorans 80:103 Zobellia galactanivorans 80:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 2 60 theme storage 232:238 arg1 polysaccharide 240:253 This storage polysaccharide 227:253 This storage polysaccharide of brown algae 227:268 This storage polysaccharide of brown algae constitutes an abundant source of carbon for marine bacteria such as Zobellia galactanivorans. 25664729 3 61 dep GH64 512:515 arg1 to 509:510 to 509:510 to 509:510 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 7 62 theme active-site 959:969 arg1 cleft 971:975 a straight active-site cleft 948:975 a straight active-site cleft 948:975 The structure of an inactive mutant of ZgLamCGH16 in complex with a thio-β-1,3-hexaglucan substrate unravelled a straight active-site cleft with three additional pockets flanking subsites -1, -2 and -3. 25664729 9 63 theme glucose 1220:1226 arg1 moiety 1228:1233 each glucose moiety 1215:1233 each glucose moiety 1215:1233 The presence of these molecules in the vicinity of the O6 hydroxyl group of each glucose moiety suggests that ZgLamCGH16 accommodates branched laminarins as substrates. 25664729 3 64 theme glycosyl 465:472 arg1 family 484:489 glycosyl hydrolase family 16 465:492 glycosyl hydrolase family 16 (GH16) 465:499 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 64 theme glycosyl 465:472 arg1 GH16 495:498 GH16 495:498 GH16 495:498 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 0 65 from characterization 27:42 arg1 galactanivorans 89:103 Zobellia galactanivorans 80:103 Zobellia galactanivorans 80:103 Structural and biochemical characterization of the laminarinase ZgLamCGH16 from Zobellia galactanivorans suggests preferred recognition of branched laminarin. 25664729 6 66 link mixed-linked 810:821 arg1 MLG 831:833 MLG 831:833 MLG 831:833 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 6 66 link mixed-linked 810:821 arg1 glucan 823:828 mixed-linked glucan 810:828 mixed-linked glucan (MLG) 810:834 This recombinant enzyme has a preferential specificity for laminarin but also a significant activity on mixed-linked glucan (MLG). 25664729 8 67 used occupied 1066:1073 arg2 pockets 1054:1060 These lateral pockets 1040:1060 These lateral pockets 1040:1060 These lateral pockets are occupied by a glycerol, an acetate ion and a chloride ion, respectively. 25664729 3 68 theme putative 420:427 arg1 β-1,3-glucanases 429:444 five putative β-1,3-glucanases 415:444 five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures 415:551 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 68 theme putative 420:427 arg1 GH64 512:515 GH64 512:515 GH64 512:515 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 25664729 3 68 theme putative 420:427 arg1 [four 446:450 [four 446:450 [four 446:450 This marine member of the Bacteroidetes possesses five putative β-1,3-glucanases [four belonging to glycosyl hydrolase family 16 (GH16) and one to GH64] with various modular architectures. 24997456 3 0 theme gross 489:493 arg1 structure 495:503 gross structure 489:503 gross structure 489:503 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 5 1 theme Human 915:919 arg1 corneas 921:927 Human corneas 915:927 Human corneas 915:927 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 1 2 gly glycoprotein 169:180 arg1 glycoprotein 169:180 a lubricating mucin-like glycoprotein 144:180 a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role 144:304 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 1 2 gly glycoprotein 169:180 arg1 Proteoglycan 106:117 Proteoglycan 4 106:119 Proteoglycan 4 (PRG4, or lubricin) 106:139 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 3 3 theme rhPRG4 539:544 arg1 glycosylations 509:522 glycosylations 509:522 glycosylations 509:522 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 3 3 theme rhPRG4 539:544 arg1 structure 495:503 gross structure 489:503 gross structure 489:503 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 11 4 contain have 1759:1762 arg2 utility 1773:1779 clinical utility 1764:1779 clinical utility 1764:1779 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 4 contain have 1759:1762 arg2 treatment 1794:1802 a topical treatment 1784:1802 a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear 1784:1890 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 4 contain have 1759:1762 arg1 rhPRG4 1748:1753 Purified rhPRG4 1739:1753 Purified rhPRG4 1739:1753 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 10 5 theme boundary 1717:1724 arg1 lubricating 1726:1736 ocular surface boundary lubricating 1702:1736 ocular surface boundary lubricating 1702:1736 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 1 6 theme mucin-like 158:167 arg1 glycoprotein 169:180 a lubricating mucin-like glycoprotein 144:180 a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role 144:304 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 1 6 theme mucin-like 158:167 arg1 Proteoglycan 106:117 Proteoglycan 4 106:119 Proteoglycan 4 (PRG4, or lubricin) 106:139 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 11 7 theme Purified 1739:1746 arg1 rhPRG4 1748:1753 Purified rhPRG4 1739:1753 Purified rhPRG4 1739:1753 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 7 8 contain had 1249:1251 arg1 rhPRG4 1242:1247 rhPRG4 1242:1247 rhPRG4 1242:1247 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 7 8 contain had 1249:1251 arg2 immunoreactivity 1253:1268 immunoreactivity 1253:1268 immunoreactivity 1253:1268 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 7 9 theme consistent 1353:1362 arg1 glycosylation 1339:1351 O-linked glycosylation 1330:1351 O-linked glycosylation consistent with that of PRG4 1330:1380 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 11 10 theme dry 1807:1809 arg1 disease 1815:1821 dry eye disease 1807:1821 dry eye disease 1807:1821 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 3 11 theme surface 572:578 arg1 ability 601:607 the ocular surface boundary lubricating ability 561:607 the ocular surface boundary lubricating ability of rhPRG4 561:617 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 1 12 theme ocular 209:214 arg1 surface 216:222 the ocular surface 205:222 the ocular surface 205:222 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 4 13 theme tandem 881:886 arg1 MS/MS 907:911 MS/MS 907:911 MS/MS 907:911 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 4 13 theme tandem 881:886 arg1 spectrometry 893:904 tandem mass spectrometry 881:904 tandem mass spectrometry (MS/MS) 881:912 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 7 14 theme O-linked 1330:1337 arg1 glycosylation 1339:1351 O-linked glycosylation 1330:1351 O-linked glycosylation consistent with that of PRG4 1330:1380 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 10 15 link O-linked 1673:1680 arg1 glycosylations 1682:1695 O-linked glycosylations 1673:1695 O-linked glycosylations 1673:1695 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 1 16 dep Proteoglycan 106:117 arg1 PRG4 122:125 PRG4 122:125 PRG4 122:125 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 1 16 dep Proteoglycan 106:117 arg1 lubricin 131:138 lubricin 131:138 lubricin 131:138 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 5 17 theme lubricating 1095:1105 arg1 ability 1107:1113 the ocular lubricating ability 1084:1113 the ocular lubricating ability of rhPRG4 to PRG4 1084:1131 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 3 18 theme lubricating 589:599 arg1 ability 601:607 the ocular surface boundary lubricating ability 561:607 the ocular surface boundary lubricating ability of rhPRG4 561:617 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 0 19 theme surface 78:84 arg1 boundary 86:93 an ocular surface boundary lubricant 68:103 an ocular surface boundary lubricant 68:103 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 0 20 theme lubricant 95:103 arg1 boundary 86:93 an ocular surface boundary lubricant 68:103 an ocular surface boundary lubricant 68:103 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 2 21 theme human 397:401 arg1 rhPRG4 409:414 rhPRG4 409:414 rhPRG4 409:414 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 2 21 theme human 397:401 arg1 PRG4 403:406 full-length recombinant human PRG4 373:406 full-length recombinant human PRG4 (rhPRG4) 373:415 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 5 22 theme human 962:966 arg1 eyelids 968:974 human eyelids 962:974 human eyelids 962:974 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 7 23 theme molecular 1298:1306 arg1 weight 1308:1313 the appropriate apparent molecular weight 1273:1313 the appropriate apparent molecular weight 1273:1313 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 4 24 theme Chinese 729:735 arg1 line 756:759 a Chinese hamster ovary cell line 727:759 a Chinese hamster ovary cell line 727:759 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 2 25 theme full-length 373:383 arg1 rhPRG4 409:414 rhPRG4 409:414 rhPRG4 409:414 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 2 25 theme full-length 373:383 arg1 PRG4 403:406 full-length recombinant human PRG4 373:406 full-length recombinant human PRG4 (rhPRG4) 373:415 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 1 26 theme lubricant 258:266 arg1 it 231:232 it 231:232 it 231:232 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 1 26 theme lubricant 258:266 arg1 boundary 249:256 a boundary lubricant 247:266 a boundary lubricant 247:266 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 7 27 theme appropriate 1277:1287 arg1 weight 1308:1313 the appropriate apparent molecular weight 1273:1313 the appropriate apparent molecular weight 1273:1313 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 4 28 theme ovary 745:749 arg1 line 756:759 a Chinese hamster ovary cell line 727:759 a Chinese hamster ovary cell line 727:759 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 10 29 theme order 1656:1660 arg1 structure 1662:1670 appropriate higher order structure 1637:1670 appropriate higher order structure 1637:1670 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 0 30 theme full-length 20:30 arg1 Proteoglycan 50:61 full-length recombinant human Proteoglycan 4 20:63 full-length recombinant human Proteoglycan 4 20:63 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 7 31 link O-linked 1330:1337 arg1 glycosylation 1339:1351 O-linked glycosylation 1330:1351 O-linked glycosylation consistent with that of PRG4 1330:1380 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 3 32 theme human 627:631 arg1 cornea-eyelid 633:645 human cornea-eyelid 627:645 human cornea-eyelid 627:645 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 10 33 theme appropriate 1637:1647 arg1 structure 1662:1670 appropriate higher order structure 1637:1670 appropriate higher order structure 1637:1670 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 3 34 dep assess 554:559 arg1 2 551:551 2 551:551 2 551:551 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 0 35 theme human 44:48 arg1 Proteoglycan 50:61 full-length recombinant human Proteoglycan 4 20:63 full-length recombinant human Proteoglycan 4 20:63 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 9 36 theme human 1550:1554 arg1 biointerfaces 1570:1582 human cornea-eyelid biointerfaces 1550:1582 human cornea-eyelid biointerfaces 1550:1582 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 4 37 theme SDS-PAGE 817:824 arg1 blotting 834:841 SDS-PAGE western blotting 817:841 SDS-PAGE western blotting 817:841 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 11 38 theme contact 1826:1832 arg1 lens 1834:1837 contact lens 1826:1837 contact lens 1826:1837 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 1 39 theme protective 290:299 arg1 role 301:304 a protective role 288:304 a protective role 288:304 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 5 40 theme 0.3-30 mm/s 1012:1022 arg1 velocities 998:1007 effective sliding velocities 980:1007 effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa 980:1059 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 3 41 theme cornea-polydimethylsiloxane 657:683 arg1 biointerfaces 692:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 8 42 theme protein 1390:1396 arg1 identity 1398:1405 rhPRG4 protein identity 1383:1405 rhPRG4 protein identity 1383:1405 rhPRG4 protein identity was confirmed by MS/MS. 24997456 2 43 theme Recent 307:312 arg1 advances 328:335 Recent technological advances 307:335 Recent technological advances 307:335 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 2 44 theme PRG4 403:406 arg1 expression 359:368 abundant expression 350:368 abundant expression of full-length recombinant human PRG4 (rhPRG4) 350:415 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 4 45 theme native 795:800 arg1 PRG4 809:812 native bovine PRG4 795:812 native bovine PRG4 795:812 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 10 46 theme surface 1709:1715 arg1 lubricating 1726:1736 ocular surface boundary lubricating 1702:1736 ocular surface boundary lubricating 1702:1736 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 7 47 contain possessed 1320:1328 arg1 rhPRG4 1242:1247 rhPRG4 1242:1247 rhPRG4 1242:1247 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 7 47 contain possessed 1320:1328 arg2 glycosylation 1339:1351 O-linked glycosylation 1330:1351 O-linked glycosylation consistent with that of PRG4 1330:1380 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 5 48 theme effective 980:988 arg1 velocities 998:1007 effective sliding velocities 980:1007 effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa 980:1059 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 11 49 theme biomaterial 1839:1849 arg1 coating 1851:1857 biomaterial coating 1839:1857 biomaterial coating 1839:1857 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 4 50 theme protein 848:854 arg1 identity 856:863 protein identity 848:863 protein identity 848:863 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 3 51 theme PDMS 686:689 arg1 biointerfaces 692:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 3 52 theme full-length 527:537 arg1 rhPRG4 539:544 full-length rhPRG4 527:544 full-length rhPRG4 527:544 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 1 53 theme lubricating 146:156 arg1 glycoprotein 169:180 a lubricating mucin-like glycoprotein 144:180 a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role 144:304 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 1 53 theme lubricating 146:156 arg1 Proteoglycan 106:117 Proteoglycan 4 106:119 Proteoglycan 4 (PRG4, or lubricin) 106:139 Proteoglycan 4 (PRG4, or lubricin) is a lubricating mucin-like glycoprotein recently discovered at the ocular surface, where it functions as a boundary lubricant and appears to play a protective role. 24997456 4 54 theme western 826:832 arg1 blotting 834:841 SDS-PAGE western blotting 817:841 SDS-PAGE western blotting 817:841 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 11 55 theme topical 1786:1792 arg1 utility 1773:1779 clinical utility 1764:1779 clinical utility 1764:1779 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 55 theme topical 1786:1792 arg1 treatment 1794:1802 a topical treatment 1784:1802 a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear 1784:1890 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 6 56 dep μg/ml 1189:1193 arg1 both 1180:1183 both 1180:1183 both 1180:1183 Samples were tested serially in PRG4, rhPRG4 (both 300 μg/ml), then saline. 24997456 2 57 theme technological 314:326 arg1 advances 328:335 Recent technological advances 307:335 Recent technological advances 307:335 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 3 58 theme ocular 565:570 arg1 ability 601:607 the ocular surface boundary lubricating ability 561:607 the ocular surface boundary lubricating ability of rhPRG4 561:617 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 11 59 theme clinical 1764:1771 arg1 utility 1773:1779 clinical utility 1764:1779 clinical utility 1764:1779 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 59 theme clinical 1764:1771 arg1 treatment 1794:1802 a topical treatment 1784:1802 a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear 1784:1890 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 60 theme eye 1811:1813 arg1 disease 1815:1821 dry eye disease 1807:1821 dry eye disease 1807:1821 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 5 61 theme physiological 1030:1042 arg1 loads 1044:1048 physiological loads 1030:1048 physiological loads of ∼15 kPa 1030:1059 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 9 62 from PDMS 1541:1544 arg1 saline 1511:1516 saline 1511:1516 saline at both human cornea - PDMS and human cornea-eyelid biointerfaces 1511:1582 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 4 63 theme mass 888:891 arg1 MS/MS 907:911 MS/MS 907:911 MS/MS 907:911 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 4 63 theme mass 888:891 arg1 spectrometry 893:904 tandem mass spectrometry 881:904 tandem mass spectrometry (MS/MS) 881:912 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 11 64 theme disease 1815:1821 arg1 utility 1773:1779 clinical utility 1764:1779 clinical utility 1764:1779 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 64 theme disease 1815:1821 arg1 treatment 1794:1802 a topical treatment 1784:1802 a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear 1784:1890 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 0 65 theme ocular 71:76 arg1 boundary 86:93 an ocular surface boundary lubricant 68:103 an ocular surface boundary lubricant 68:103 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 3 66 theme boundary 580:587 arg1 ability 601:607 the ocular surface boundary lubricating ability 561:607 the ocular surface boundary lubricating ability of rhPRG4 561:617 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 9 67 from cornea 1532:1537 arg1 saline 1511:1516 saline 1511:1516 saline at both human cornea - PDMS and human cornea-eyelid biointerfaces 1511:1582 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 3 68 gly glycosylations 509:522 arg1 rhPRG4 539:544 full-length rhPRG4 527:544 full-length rhPRG4 527:544 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 5 69 theme ∼15 kPa 1053:1059 arg1 loads 1044:1048 physiological loads 1030:1048 physiological loads of ∼15 kPa 1030:1059 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 5 70 theme rhPRG4 1118:1123 arg1 ability 1107:1113 the ocular lubricating ability 1084:1113 the ocular lubricating ability of rhPRG4 to PRG4 1084:1131 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 11 71 dep disease 1815:1821 arg1 coating 1851:1857 biomaterial coating 1839:1857 biomaterial coating 1839:1857 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 4 72 theme cell 751:754 arg1 line 756:759 a Chinese hamster ovary cell line 727:759 a Chinese hamster ovary cell line 727:759 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 7 73 with consistent 1353:1362 arg1 that 1369:1372 that 1369:1372 that 1369:1372 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 7 74 theme apparent 1289:1296 arg1 weight 1308:1313 the appropriate apparent molecular weight 1273:1313 the appropriate apparent molecular weight 1273:1313 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 2 75 theme recombinant 385:395 arg1 rhPRG4 409:414 rhPRG4 409:414 rhPRG4 409:414 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 2 75 theme recombinant 385:395 arg1 PRG4 403:406 full-length recombinant human PRG4 373:406 full-length recombinant human PRG4 (rhPRG4) 373:415 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 5 76 theme ocular 1088:1093 arg1 ability 1107:1113 the ocular lubricating ability 1084:1113 the ocular lubricating ability of rhPRG4 to PRG4 1084:1131 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 9 77 theme human 1526:1530 arg1 cornea 1532:1537 human cornea 1526:1537 human cornea 1526:1537 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 0 78 theme recombinant 32:42 arg1 Proteoglycan 50:61 full-length recombinant human Proteoglycan 4 20:63 full-length recombinant human Proteoglycan 4 20:63 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 3 79 dep structure 495:503 arg1 the 485:487 the 485:487 the 485:487 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 9 80 from biointerfaces 1570:1582 arg1 saline 1511:1516 saline 1511:1516 saline at both human cornea - PDMS and human cornea-eyelid biointerfaces 1511:1582 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 4 81 theme hamster 737:743 arg1 line 756:759 a Chinese hamster ovary cell line 727:759 a Chinese hamster ovary cell line 727:759 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 10 82 theme higher 1649:1654 arg1 structure 1662:1670 appropriate higher order structure 1637:1670 appropriate higher order structure 1637:1670 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 0 83 theme Proteoglycan 50:61 arg1 Characterization 0:15 Characterization 0:15 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 0:104 Characterization of full-length recombinant human Proteoglycan 4 as an ocular surface boundary lubricant. 24997456 5 84 theme sliding 990:996 arg1 velocities 998:1007 effective sliding velocities 980:1007 effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa 980:1059 Human corneas were articulated against PDMS or human eyelids, at effective sliding velocities of 0.3-30 mm/s under physiological loads of ∼15 kPa, to assess and compare the ocular lubricating ability of rhPRG4 to PRG4. 24997456 3 85 theme rhPRG4 612:617 arg1 ability 601:607 the ocular surface boundary lubricating ability 561:607 the ocular surface boundary lubricating ability of rhPRG4 561:617 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 4 86 theme bovine 802:807 arg1 PRG4 809:812 native bovine PRG4 795:812 native bovine PRG4 795:812 rhPRG4 expressed by a Chinese hamster ovary cell line was characterized and compared to native bovine PRG4 by SDS-PAGE western blotting, and protein identity was assessed by tandem mass spectrometry (MS/MS). 24997456 3 87 theme study 441:445 arg1 objectives 422:431 The objectives 418:431 The objectives of this study 418:445 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 9 88 theme cornea-eyelid 1556:1568 arg1 biointerfaces 1570:1582 human cornea-eyelid biointerfaces 1550:1582 human cornea-eyelid biointerfaces 1550:1582 Both PRG4 and rhPRG4 significantly, and similarly, reduced friction compared to saline at both human cornea - PDMS and human cornea-eyelid biointerfaces. 24997456 3 89 theme human 651:655 arg1 biointerfaces 692:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 human cornea-polydimethylsiloxane (PDMS) biointerfaces 651:704 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 10 90 theme O-linked 1673:1680 arg1 glycosylations 1682:1695 O-linked glycosylations 1673:1695 O-linked glycosylations 1673:1695 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24997456 11 91 theme lens 1834:1837 arg1 utility 1773:1779 clinical utility 1764:1779 clinical utility 1764:1779 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 11 91 theme lens 1834:1837 arg1 treatment 1794:1802 a topical treatment 1784:1802 a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear 1784:1890 Purified rhPRG4 may have clinical utility as a topical treatment of dry eye disease or contact lens biomaterial coating to promote more comfortable wear. 24997456 7 92 theme Western 1210:1216 arg1 blotting 1218:1225 Western blotting 1210:1225 Western blotting 1210:1225 Western blotting indicated that rhPRG4 had immunoreactivity at the appropriate apparent molecular weight, and possessed O-linked glycosylation consistent with that of PRG4. 24997456 3 93 dep characterize 472:483 arg1 1 455:455 1 455:455 1 455:455 The objectives of this study were to 1) biochemically characterize the gross structure and glycosylations of full-length rhPRG4, and 2) assess the ocular surface boundary lubricating ability of rhPRG4 at both human cornea-eyelid and human cornea-polydimethylsiloxane (PDMS) biointerfaces. 24997456 8 94 theme rhPRG4 1383:1388 arg1 identity 1398:1405 rhPRG4 protein identity 1383:1405 rhPRG4 protein identity 1383:1405 rhPRG4 protein identity was confirmed by MS/MS. 24997456 2 95 theme abundant 350:357 arg1 expression 359:368 abundant expression 350:368 abundant expression of full-length recombinant human PRG4 (rhPRG4) 350:415 Recent technological advances have enabled abundant expression of full-length recombinant human PRG4 (rhPRG4). 24997456 10 96 theme ocular 1702:1707 arg1 lubricating 1726:1736 ocular surface boundary lubricating 1702:1736 ocular surface boundary lubricating 1702:1736 In conclusion, the rhPRG4 studied here demonstrated appropriate higher order structure, O-linked glycosylations, and ocular surface boundary lubricating. 24927272 0 0 theme glycosaminoglycans 91:108 arg1 attachment 77:86 attachment 77:86 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 0:182 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 3 1 theme linker 657:662 arg1 repeat 664:669 the common (G4S) 4 linker repeat 638:669 the common (G4S) 4 linker repeat 638:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 4 2 contain containing 750:759 arg2 Xyl 777:779 at least a core Xyl 761:779 at least a core Xyl 761:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 4 2 contain containing 750:759 arg1 glycans 738:744 xylose-based glycans 725:744 xylose-based glycans all containing at least a core Xyl 725:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 6 3 with glycans 1080:1086 arg1 addition 1100:1107 a mass addition 1093:1107 a mass addition of 79.9 Da 1093:1118 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 3 4 theme G4S 650:652 arg1 repeat 664:669 the common (G4S) 4 linker repeat 638:669 the common (G4S) 4 linker repeat 638:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 0 5 with glycosaminoglycans 91:108 arg1 phosphorylation 139:153 phosphorylation 139:153 phosphorylation 139:153 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 5 6 dep Xyl-Gal 886:892 arg1 e.g. 880:883 e.g. 880:883 e.g. 880:883 Commonly observed glycan structures include GAG-related di-, tri-, tetra-, and penta-saccharides (e.g., Xyl-Gal, Xyl-Gal-Gal, Xyl-Gal-Gal-GlcA, and Xyl-Gal-Gal-GlcA-HexNAc), as well as Xyl-Gal-Neu5Ac. 24927272 0 7 theme other 114:118 arg1 intermediates 120:132 other intermediates 114:132 other intermediates 114:132 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 0 8 from O-glycosylation 0:14 arg1 proteins 67:74 recombinant Fc-fusion proteins 45:74 recombinant Fc-fusion proteins 45:74 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 7 9 theme due 1300:1302 arg1 moiety 1293:1298 the HexNAc moiety 1282:1298 the HexNAc moiety due to non-reactivity to alkaline phosphatase 1282:1344 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 6 10 theme alkaline 993:1000 arg1 phosphatase 1002:1012 alkaline phosphatase 993:1012 alkaline phosphatase 993:1012 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 0 11 from subunit 175:181 arg1 attachment 77:86 attachment 77:86 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 0:182 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 8 12 theme linker 1445:1450 arg1 peptide 1452:1458 the linker peptide 1441:1458 the linker peptide 1441:1458 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 1 13 theme linker 276:281 arg1 sequence 283:290 the linker sequence 272:290 the linker sequence of a recombinant Fc fusion protein 272:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 2 14 theme minor 514:518 arg1 intermediates 520:532 other minor intermediates 508:532 other minor intermediates 508:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 7 15 theme phosphorylated 1197:1210 arg1 pentasaccharides 1216:1231 phosphorylated GAG pentasaccharides 1197:1231 phosphorylated GAG pentasaccharides 1197:1231 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 6 16 theme phosphatase 1002:1012 arg1 treatment 1027:1035 alkaline phosphatase or sialidase treatment 993:1035 alkaline phosphatase or sialidase treatment combined with CID fragmentation 993:1067 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 4 17 theme glycans 738:744 arg1 population 711:720 a heterogenous population 696:720 a heterogenous population of xylose-based glycans all containing at least a core Xyl 696:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 1 18 theme xylose-based 186:197 arg1 GAG 218:220 GAG 218:220 GAG 218:220 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 1 18 theme xylose-based 186:197 arg1 glycosaminoglycan 199:215 A xylose-based glycosaminoglycan 184:215 A xylose-based glycosaminoglycan (GAG) core 184:226 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 0 19 theme intermediates 120:132 arg1 attachment 77:86 attachment 77:86 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 0:182 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 8 20 theme low 1516:1518 arg1 levels 1520:1525 very low levels 1511:1525 very low levels 1511:1525 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 4 21 theme xylose-based 725:736 arg1 glycans 738:744 xylose-based glycans 725:744 xylose-based glycans all containing at least a core Xyl 725:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 1 22 theme glycosaminoglycan 199:215 arg1 core 223:226 A xylose-based glycosaminoglycan (GAG) core 184:226 A xylose-based glycosaminoglycan (GAG) core 184:226 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 8 23 theme multiple 1479:1486 arg1 additions 1488:1496 multiple additions 1479:1496 multiple additions of xylose at very low levels 1479:1525 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 0 24 with intermediates 120:132 arg1 phosphorylation 139:153 phosphorylation 139:153 phosphorylation 139:153 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 6 25 theme Da 1117:1118 arg1 addition 1100:1107 a mass addition 1093:1107 a mass addition of 79.9 Da 1093:1118 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 2 26 theme acidic 378:383 arg1 residue 385:391 an upstream acidic residue 366:391 an upstream acidic residue 366:391 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 26 theme acidic 378:383 arg1 sequence 339:346 The linker sequence 328:346 The linker sequence 328:346 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 26 theme acidic 378:383 arg1 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 26 theme acidic 378:383 arg1 substrate 411:419 a substrate 409:419 a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates 409:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 1 27 from residue 261:267 arg1 sequence 283:290 the linker sequence 272:290 the linker sequence of a recombinant Fc fusion protein 272:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 2 28 theme upstream 369:376 arg1 residue 385:391 an upstream acidic residue 366:391 an upstream acidic residue 366:391 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 28 theme upstream 369:376 arg1 sequence 339:346 The linker sequence 328:346 The linker sequence 328:346 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 28 theme upstream 369:376 arg1 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 28 theme upstream 369:376 arg1 substrate 411:419 a substrate 409:419 a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates 409:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 0 29 theme linkers 34:40 arg1 O-glycosylation 0:14 O-glycosylation 0:14 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 0:182 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 6 30 theme xylose 1168:1173 arg1 result 1143:1148 a result 1141:1148 a result of phosphorylated xylose 1141:1173 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 6 30 theme xylose 1168:1173 arg1 glycans 1080:1086 low-level glycans 1070:1086 low-level glycans with a mass addition of 79.9 Da 1070:1118 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 4 31 theme heterogenous 698:709 arg1 population 711:720 a heterogenous population 696:720 a heterogenous population of xylose-based glycans all containing at least a core Xyl 696:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 4 32 theme core 772:775 arg1 Xyl 777:779 at least a core Xyl 761:779 at least a core Xyl 761:779 This linker resulted in a heterogenous population of xylose-based glycans all containing at least a core Xyl. 24927272 0 33 theme glycine-serine 19:32 arg1 linkers 34:40 glycine-serine linkers 19:40 glycine-serine linkers 19:40 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 3 34 theme Fc 622:623 arg1 domain 625:630 an IgG Fc domain 615:630 an IgG Fc domain using the common (G4S) 4 linker repeat 615:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 6 35 theme CID 1051:1053 arg1 fragmentation 1055:1067 CID fragmentation 1051:1067 CID fragmentation 1051:1067 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 5 36 theme observed 791:798 arg1 structures 807:816 Commonly observed glycan structures 782:816 Commonly observed glycan structures 782:816 Commonly observed glycan structures include GAG-related di-, tri-, tetra-, and penta-saccharides (e.g., Xyl-Gal, Xyl-Gal-Gal, Xyl-Gal-Gal-GlcA, and Xyl-Gal-Gal-GlcA-HexNAc), as well as Xyl-Gal-Neu5Ac. 24927272 0 37 theme recombinant 45:55 arg1 proteins 67:74 recombinant Fc-fusion proteins 45:74 recombinant Fc-fusion proteins 45:74 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 1 38 theme recombinant 297:307 arg1 protein 319:325 a recombinant Fc fusion protein 295:325 a recombinant Fc fusion protein 295:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 0 39 theme xylose 162:167 arg1 subunit 175:181 the xylose sugar subunit 158:181 the xylose sugar subunit 158:181 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 3 40 theme domain 625:630 arg1 C-terminus 601:610 the C-terminus 597:610 the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat 597:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 0 41 gly O-glycosylation 0:14 arg1 linkers 34:40 glycine-serine linkers 19:40 glycine-serine linkers 19:40 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 0 41 gly O-glycosylation 0:14 arg1 proteins 67:74 recombinant Fc-fusion proteins 45:74 recombinant Fc-fusion proteins 45:74 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 6 42 theme sialidase 1017:1025 arg1 treatment 1027:1035 alkaline phosphatase or sialidase treatment 993:1035 alkaline phosphatase or sialidase treatment combined with CID fragmentation 993:1067 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 5 43 theme glycan 800:805 arg1 structures 807:816 Commonly observed glycan structures 782:816 Commonly observed glycan structures 782:816 Commonly observed glycan structures include GAG-related di-, tri-, tetra-, and penta-saccharides (e.g., Xyl-Gal, Xyl-Gal-Gal, Xyl-Gal-Gal-GlcA, and Xyl-Gal-Gal-GlcA-HexNAc), as well as Xyl-Gal-Neu5Ac. 24927272 2 44 theme major 462:466 arg1 glycan 468:473 a major glycan 460:473 a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates 460:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 1 45 theme Fc 309:310 arg1 protein 319:325 a recombinant Fc fusion protein 295:325 a recombinant Fc fusion protein 295:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 8 46 from levels 1520:1525 arg1 additions 1488:1496 multiple additions 1479:1496 multiple additions of xylose at very low levels 1479:1525 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 2 47 theme other 508:512 arg1 intermediates 520:532 other minor intermediates 508:532 other minor intermediates 508:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 1 48 theme fusion 312:317 arg1 protein 319:325 a recombinant Fc fusion protein 295:325 a recombinant Fc fusion protein 295:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 3 49 theme IgG 618:620 arg1 domain 625:630 an IgG Fc domain 615:630 an IgG Fc domain using the common (G4S) 4 linker repeat 615:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 3 50 theme unrelated 566:574 arg1 protein 576:582 an unrelated protein 563:582 an unrelated protein 563:582 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 0 51 attach attachment 77:86 arg2 intermediates 120:132 other intermediates 114:132 other intermediates 114:132 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 0 51 attach attachment 77:86 arg2 glycosaminoglycans 91:108 glycosaminoglycans 91:108 glycosaminoglycans 91:108 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 0 51 attach attachment 77:86 arg3 subunit 175:181 the xylose sugar subunit 158:181 the xylose sugar subunit 158:181 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 6 52 theme mass 1095:1098 arg1 addition 1100:1107 a mass addition 1093:1107 a mass addition of 79.9 Da 1093:1118 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 0 53 theme Fc-fusion 57:65 arg1 proteins 67:74 recombinant Fc-fusion proteins 45:74 recombinant Fc-fusion proteins 45:74 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 1 54 theme protein 319:325 arg1 sequence 283:290 the linker sequence 272:290 the linker sequence of a recombinant Fc fusion protein 272:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 24927272 0 55 theme sugar 169:173 arg1 subunit 175:181 the xylose sugar subunit 158:181 the xylose sugar subunit 158:181 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 3 56 theme common 642:647 arg1 repeat 664:669 the common (G4S) 4 linker repeat 638:669 the common (G4S) 4 linker repeat 638:669 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 3 57 theme protein 576:582 arg1 portion 552:558 a portion 550:558 a portion of an unrelated protein 550:582 In this paper, a portion of an unrelated protein was fused to the C-terminus of an IgG Fc domain using the common (G4S) 4 linker repeat. 24927272 7 58 theme HexNAc 1286:1291 arg1 moiety 1293:1298 the HexNAc moiety 1282:1298 the HexNAc moiety due to non-reactivity to alkaline phosphatase 1282:1344 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 6 59 theme phosphorylated 1153:1166 arg1 xylose 1168:1173 phosphorylated xylose 1153:1173 phosphorylated xylose 1153:1173 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 2 60 theme linker 332:337 arg1 sequence 339:346 The linker sequence 328:346 The linker sequence 328:346 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 60 theme linker 332:337 arg1 residue 385:391 an upstream acidic residue 366:391 an upstream acidic residue 366:391 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 60 theme linker 332:337 arg1 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 G-S-G-G-G-G 349:359 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 2 60 theme linker 332:337 arg1 substrate 411:419 a substrate 409:419 a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates 409:532 The linker sequence, G-S-G-G-G-G, and an upstream acidic residue were serving as a substrate for O-xylosyltransferase, resulting in a major glycan composed of Xyl-Gal-Gal-GlcA and other minor intermediates. 24927272 7 61 theme alkaline 1325:1332 arg1 phosphatase 1334:1344 alkaline phosphatase 1325:1344 alkaline phosphatase 1325:1344 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 8 62 theme G-S-G 1414:1418 arg1 motifs 1429:1434 the three G-S-G sequence motifs 1404:1434 the three G-S-G sequence motifs 1404:1434 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 8 63 theme xylose 1351:1356 arg1 moiety 1358:1363 The xylose moiety 1347:1363 The xylose moiety 1347:1363 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 8 64 theme sequence 1420:1427 arg1 motifs 1429:1434 the three G-S-G sequence motifs 1404:1434 the three G-S-G sequence motifs 1404:1434 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 7 65 dep sulfated 1245:1252 arg1 Da 1265:1266 79.9 Da 1260:1266 also 79.9 Da 1255:1266 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 7 66 theme pentasaccharides 1216:1231 arg1 quantity 1185:1192 A minute quantity 1176:1192 A minute quantity of phosphorylated GAG pentasaccharides 1176:1231 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 6 67 theme low-level 1070:1078 arg1 result 1143:1148 a result 1141:1148 a result of phosphorylated xylose 1141:1173 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 6 67 theme low-level 1070:1078 arg1 glycans 1080:1086 low-level glycans 1070:1086 low-level glycans with a mass addition of 79.9 Da 1070:1118 Following alkaline phosphatase or sialidase treatment combined with CID fragmentation, low-level glycans with a mass addition of 79.9 Da were confirmed to be a result of phosphorylated xylose. 24927272 5 68 theme GAG-related 826:836 arg1 di- 838:840 di- 838:840 di- 838:840 Commonly observed glycan structures include GAG-related di-, tri-, tetra-, and penta-saccharides (e.g., Xyl-Gal, Xyl-Gal-Gal, Xyl-Gal-Gal-GlcA, and Xyl-Gal-Gal-GlcA-HexNAc), as well as Xyl-Gal-Neu5Ac. 24927272 8 69 theme xylose 1501:1506 arg1 additions 1488:1496 multiple additions 1479:1496 multiple additions of xylose at very low levels 1479:1525 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 7 70 theme minute 1178:1183 arg1 quantity 1185:1192 A minute quantity 1176:1192 A minute quantity of phosphorylated GAG pentasaccharides 1176:1231 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 8 71 theme motifs 1429:1434 arg1 one 1397:1399 one 1397:1399 one 1397:1399 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 8 71 theme motifs 1429:1434 arg1 motifs 1429:1434 the three G-S-G sequence motifs 1404:1434 the three G-S-G sequence motifs 1404:1434 The xylose moiety may be randomly incorporated in one of the three G-S-G sequence motifs; and the linker peptide shows evidence for multiple additions of xylose at very low levels. 24927272 0 72 dep O-glycosylation 0:14 arg1 attachment 77:86 attachment 77:86 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 0:182 O-glycosylation of glycine-serine linkers in recombinant Fc-fusion proteins: attachment of glycosaminoglycans and other intermediates with phosphorylation at the xylose sugar subunit. 24927272 7 73 theme GAG 1212:1214 arg1 pentasaccharides 1216:1231 phosphorylated GAG pentasaccharides 1197:1231 phosphorylated GAG pentasaccharides 1197:1231 A minute quantity of phosphorylated GAG pentasaccharides may also be sulfated (also 79.9 Da), possibly at the HexNAc moiety due to non-reactivity to alkaline phosphatase. 24927272 1 74 theme Ser 257:259 arg1 residue 261:267 a Ser residue 255:267 a Ser residue in the linker sequence of a recombinant Fc fusion protein 255:325 A xylose-based glycosaminoglycan (GAG) core was recently identified at a Ser residue in the linker sequence of a recombinant Fc fusion protein. 27286768 4 0 theme N-linked 1024:1031 arg1 glycans 1033:1039 the N-linked glycans 1020:1039 the N-linked glycans analysis 1020:1048 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 3 1 theme graphitized 738:748 arg1 pore 757:760 graphitized carbon pore 738:760 graphitized carbon pore 738:760 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 5 2 theme @ 1159:1159 arg1 ZIF-8 1160:1164 the C-magG@ZIF-8 1149:1164 the C-magG@ZIF-8 1149:1164 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 2 3 theme C-magG 579:584 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 3 theme C-magG 579:584 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 3 4 theme large 725:729 arg1 sum 731:733 a large sum 723:733 a large sum of graphitized carbon pore 723:760 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 4 5 theme excellent 900:908 arg1 selectivity 910:920 excellent selectivity 900:920 excellent selectivity 900:920 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 1 6 theme selective 188:196 arg1 enrichment 198:207 The highly selective enrichment 177:207 The highly selective enrichment of N-linked glycans from complex biological sample 177:258 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 1 6 theme selective 188:196 arg1 important 274:282 important 274:282 important 274:282 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 1 7 theme challenging 288:298 arg1 task 300:303 challenging task 288:303 challenging task due to the ultra-low abundance 288:334 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 4 8 theme incredible 962:971 arg1 exclusion 978:986 incredible size exclusion 962:986 incredible size exclusion 962:986 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 6 9 theme mass 1347:1350 arg1 analysis 1365:1372 mass spectrometry analysis 1347:1372 mass spectrometry analysis 1347:1372 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 6 10 theme MOFs-functionalized 1261:1279 arg1 composites 1281:1290 the MOFs-functionalized composites 1257:1290 the MOFs-functionalized composites for large-scale characterization of glycoproteomics 1257:1342 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 0 11 theme high-effective 109:122 arg1 extraction 124:133 high-effective extraction 109:133 high-effective extraction 109:133 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 2 12 theme smart 615:619 arg1 process 621:627 a smart process 613:627 a smart process 613:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 4 13 theme glycans 1033:1039 arg1 analysis 1041:1048 the N-linked glycans analysis 1020:1048 the N-linked glycans analysis 1020:1048 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 1 14 theme due 305:307 arg1 task 300:303 challenging task 288:303 challenging task due to the ultra-low abundance 288:334 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 2 15 theme nanoporous 540:549 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 15 theme nanoporous 540:549 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 3 16 theme remarkable 763:772 arg1 biocompatibility 774:789 remarkable biocompatibility 763:789 remarkable biocompatibility 763:789 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 1 17 link N-linked 212:219 arg1 glycans 221:227 N-linked glycans 212:227 N-linked glycans 212:227 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 6 18 theme smart 1203:1207 arg1 strategy 1209:1216 our smart strategy 1199:1216 our smart strategy 1199:1216 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 0 19 link N-linked 159:166 arg1 glycans 168:174 the N-linked glycans 155:174 the N-linked glycans 155:174 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 2 20 theme magnetic 531:538 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 20 theme magnetic 531:538 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 3 21 theme carbon 750:755 arg1 pore 757:760 graphitized carbon pore 738:760 graphitized carbon pore 738:760 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 4 22 theme @ 872:872 arg1 composites 879:888 the C-magG@ZIF-8 composites 862:888 the C-magG@ZIF-8 composites 862:888 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 0 23 theme metal-organic 10:22 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 24 theme pore 757:760 arg1 responsiveness 707:720 strong magnetic responsiveness 691:720 strong magnetic responsiveness 691:720 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 24 theme pore 757:760 arg1 biocompatibility 774:789 remarkable biocompatibility 763:789 remarkable biocompatibility 763:789 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 24 theme pore 757:760 arg1 sum 731:733 a large sum 723:733 a large sum of graphitized carbon pore 723:760 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 24 theme pore 757:760 arg1 area 818:821 large specific surface area 795:821 large specific surface area 795:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 2 25 theme -functionalized 515:529 arg1 MOFs 510:513 MOFs 510:513 MOFs 510:513 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 25 theme -functionalized 515:529 arg1 frameworks 498:507 a novel metal-organic frameworks 476:507 a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 476:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 6 26 theme glycoproteomics 1328:1342 arg1 characterization 1308:1323 large-scale characterization 1296:1323 large-scale characterization of glycoproteomics 1296:1342 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 1 27 theme N-linked 212:219 arg1 glycans 221:227 N-linked glycans 212:227 N-linked glycans 212:227 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 0 28 theme Versatile 0:8 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 29 theme large 795:799 arg1 area 818:821 large specific surface area 795:821 large specific surface area 795:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 6 30 theme large-scale 1296:1306 arg1 characterization 1308:1323 large-scale characterization 1296:1323 large-scale characterization of glycoproteomics 1296:1342 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 4 31 theme ZIF-8 873:877 arg1 composites 879:888 the C-magG@ZIF-8 composites 862:888 the C-magG@ZIF-8 composites 862:888 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 1 32 theme glycans 221:227 arg1 enrichment 198:207 The highly selective enrichment 177:207 The highly selective enrichment of N-linked glycans from complex biological sample 177:258 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 1 32 theme glycans 221:227 arg1 important 274:282 important 274:282 important 274:282 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 0 33 theme magnetic 49:56 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 34 theme specific 801:808 arg1 area 818:821 large specific surface area 795:821 large specific surface area 795:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 4 35 link N-linked 1024:1031 arg1 glycans 1033:1039 the N-linked glycans 1020:1039 the N-linked glycans analysis 1020:1048 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 6 36 theme spectrometry 1352:1363 arg1 analysis 1365:1372 mass spectrometry analysis 1347:1372 mass spectrometry analysis 1347:1372 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 4 37 dep sensitivity 926:936 arg1 ability 988:994 ability 988:994 ability 988:994 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 0 38 theme framework-functionalized 24:47 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 1 39 theme ultra-low 316:324 arg1 abundance 326:334 the ultra-low abundance 312:334 the ultra-low abundance 312:334 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 2 40 theme @ 585:585 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 40 theme @ 585:585 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 5 41 theme normal 1117:1122 arg1 serum 1130:1134 the normal human serum 1113:1134 the normal human serum treated with the C-magG@ZIF-8 1113:1164 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 2 42 theme metal-organic 484:496 arg1 MOFs 510:513 MOFs 510:513 MOFs 510:513 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 42 theme metal-organic 484:496 arg1 frameworks 498:507 a novel metal-organic frameworks 476:507 a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 476:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 1 43 theme complex 234:240 arg1 sample 253:258 complex biological sample 234:258 complex biological sample 234:258 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 0 44 theme nanoporous 67:76 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 0 45 theme glycans 168:174 arg1 extraction 124:133 high-effective extraction 109:133 high-effective extraction 109:133 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 0 45 theme glycans 168:174 arg1 purification 139:150 purification 139:150 purification 139:150 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 46 theme strong 691:696 arg1 responsiveness 707:720 strong magnetic responsiveness 691:720 strong magnetic responsiveness 691:720 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 1 47 theme biological 242:251 arg1 sample 253:258 complex biological sample 234:258 complex biological sample 234:258 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 0 48 theme graphene 58:65 arg1 composites 78:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites 0:87 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 0 49 theme N-linked 159:166 arg1 glycans 168:174 the N-linked glycans 155:174 the N-linked glycans 155:174 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 50 theme unique 663:668 arg1 responsiveness 707:720 strong magnetic responsiveness 691:720 strong magnetic responsiveness 691:720 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 50 theme unique 663:668 arg1 biocompatibility 774:789 remarkable biocompatibility 763:789 remarkable biocompatibility 763:789 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 50 theme unique 663:668 arg1 properties 670:679 the unique properties 659:679 the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area 659:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 50 theme unique 663:668 arg1 sum 731:733 a large sum 723:733 a large sum of graphitized carbon pore 723:760 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 3 50 theme unique 663:668 arg1 area 818:821 large specific surface area 795:821 large specific surface area 795:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 0 51 dep composites 78:87 arg1 matrix 98:103 As deft matrix 90:103 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 2 52 theme novel 478:482 arg1 MOFs 510:513 MOFs 510:513 MOFs 510:513 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 52 theme novel 478:482 arg1 frameworks 498:507 a novel metal-organic frameworks 476:507 a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 476:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 1 53 theme complicated 337:347 arg1 structures 349:358 complicated structures 337:358 complicated structures 337:358 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 4 54 theme good 939:942 arg1 recyclability 944:956 good recyclability 939:956 good recyclability 939:956 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 4 55 theme C-magG 866:871 arg1 composites 879:888 the C-magG@ZIF-8 composites 862:888 the C-magG@ZIF-8 composites 862:888 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 1 56 from sample 253:258 arg1 enrichment 198:207 The highly selective enrichment 177:207 The highly selective enrichment of N-linked glycans from complex biological sample 177:258 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 1 56 from sample 253:258 arg1 important 274:282 important 274:282 important 274:282 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 5 57 link 48 N-linked 1064:1074 arg1 glycans 1076:1082 48 N-linked glycans 1064:1082 48 N-linked glycans 1064:1082 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 3 58 theme surface 810:816 arg1 area 818:821 large specific surface area 795:821 large specific surface area 795:821 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 4 59 theme size 973:976 arg1 exclusion 978:986 incredible size exclusion 962:986 incredible size exclusion 962:986 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 4 60 theme unique 843:848 arg1 properties 850:859 these unique properties 837:859 these unique properties 837:859 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 2 61 dep -functionalized 515:529 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 61 dep -functionalized 515:529 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 5 62 theme 48 N-linked 1064:1074 arg1 glycans 1076:1082 48 N-linked glycans 1064:1082 48 N-linked glycans 1064:1082 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 4 63 theme properties 850:859 arg1 virtue 827:832 virtue 827:832 virtue of these unique properties 827:859 By virtue of these unique properties, the C-magG@ZIF-8 composites displayed excellent selectivity and sensitivity, good recyclability and incredible size exclusion ability (roughly 2000 times) in the N-linked glycans analysis. 27286768 3 64 theme magnetic 698:705 arg1 responsiveness 707:720 strong magnetic responsiveness 691:720 strong magnetic responsiveness 691:720 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 5 65 theme human 1124:1128 arg1 serum 1130:1134 the normal human serum 1113:1134 the normal human serum treated with the C-magG@ZIF-8 1113:1164 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 0 66 theme deft 93:96 arg1 matrix 98:103 As deft matrix 90:103 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 3 67 theme obtained 634:641 arg1 materials 643:651 The obtained materials 630:651 The obtained materials 630:651 The obtained materials enjoy the unique properties including strong magnetic responsiveness, a large sum of graphitized carbon pore, remarkable biocompatibility and large specific surface area. 27286768 0 68 theme As 90:91 arg1 matrix 98:103 As deft matrix 90:103 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 0:175 Versatile metal-organic framework-functionalized magnetic graphene nanoporous composites: As deft matrix for high-effective extraction and purification of the N-linked glycans. 27286768 1 69 theme strong 364:369 arg1 ion 371:373 strong ion 364:373 strong ion 364:373 The highly selective enrichment of N-linked glycans from complex biological sample is still very important but challenging task due to the ultra-low abundance, complicated structures and strong ion suppress effect caused by distractors such as proteins, peptides and salts. 27286768 6 70 theme new 1225:1227 arg1 possibilities 1229:1241 new possibilities 1225:1241 new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis 1225:1372 There is reason to believe that our smart strategy offers new possibilities for preparing the MOFs-functionalized composites for large-scale characterization of glycoproteomics by mass spectrometry analysis. 27286768 5 71 theme C-magG 1153:1158 arg1 ZIF-8 1160:1164 the C-magG@ZIF-8 1149:1164 the C-magG@ZIF-8 1149:1164 Furthermore, 48 N-linked glycans were clearly identified from the normal human serum treated with the C-magG@ZIF-8. 27286768 2 72 theme carbon-graphene 551:565 arg1 ZIF-8 586:590 C-magG@ZIF-8 579:590 C-magG@ZIF-8 579:590 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 27286768 2 72 theme carbon-graphene 551:565 arg1 composites 567:576 magnetic nanoporous carbon-graphene composites 531:576 magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process 531:627 Here, we firstly present a novel metal-organic frameworks (MOFs)-functionalized magnetic nanoporous carbon-graphene composites (C-magG@ZIF-8) synthesized through a smart process. 28934626 3 0 gly glycosylation 539:551 arg1 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 0 gly glycosylation 539:551 arg1 group 577:581 the primary hydroxyl group 556:581 the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 556:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 5 1 theme promising 819:827 arg1 results 829:835 promising results 819:835 promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1) 819:933 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 1 2 theme various 175:181 arg1 positions 183:191 various positions 175:191 various positions 175:191 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 6 3 theme sialyl 1006:1011 arg1 donor 1013:1017 sialyl donor 1006:1017 sialyl donor 1006:1017 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 4 from choice 214:219 arg1 donors 252:257 sialyl donors 245:257 sialyl donors 245:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 1 5 contain containing 130:139 arg2 groups 165:170 O-trifluoroacetyl (TFA) groups 141:170 O-trifluoroacetyl (TFA) groups 141:170 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 1 5 contain containing 130:139 arg1 donors 123:128 novel sialyl donors 110:128 novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions 110:191 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 2 6 theme bond 354:357 arg1 acceptors 359:367 hydrogen bond acceptors 345:367 hydrogen bond acceptors 345:367 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 4 7 theme sialylation 769:779 arg1 outcome 758:764 the outcome 754:764 the outcome of sialylation 754:779 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 6 8 theme related 1217:1223 arg1 effects 1209:1215 intramolecular substituent effects 1182:1215 intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups 1182:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 3 9 theme hydroxyl 568:575 arg1 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 9 theme hydroxyl 568:575 arg1 group 577:581 the primary hydroxyl group 556:581 the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 556:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 10 theme group 577:581 arg1 glycosylation 539:551 the model glycosylation 529:551 the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 529:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 11 theme primary 560:566 arg1 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 11 theme primary 560:566 arg1 group 577:581 the primary hydroxyl group 556:581 the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 556:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 6 12 theme effects 1209:1215 arg1 influence 1133:1141 the influence 1129:1141 the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups 1129:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 3 13 with comparison 637:646 arg1 donors 660:665 sialyl donors 653:665 sialyl donors without O-TFA groups 653:686 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 14 theme glycosyl 496:503 arg1 donors 505:510 These glycosyl donors 490:510 These glycosyl donors 490:510 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 6 15 theme further 1089:1095 arg1 studies 1097:1103 further studies 1089:1103 further studies 1089:1103 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 3 16 theme model 533:537 arg1 glycosylation 539:551 the model glycosylation 529:551 the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 529:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 2 17 from groups 235:240 arg1 donors 252:257 sialyl donors 245:257 sialyl donors 245:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 4 18 theme O-TFA 705:709 arg1 groups 711:716 O-TFA groups 705:716 O-TFA groups 705:716 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 5 19 theme sialyl 790:795 arg1 donors 797:802 Several sialyl donors 782:802 Several sialyl donors studied 782:810 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 6 20 theme acyl 978:981 arg1 groups 996:1001 varying acyl O-protecting groups 970:1001 varying acyl O-protecting groups in sialyl donor 970:1017 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 21 theme hydrogen 1161:1168 arg1 bonding 1170:1176 intermolecular hydrogen bonding 1146:1176 intermolecular hydrogen bonding 1146:1176 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 5 22 dep 15:1 929:932 arg1 to 926:927 to 926:927 to 926:927 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 2 23 from donors 252:257 arg1 choice 214:219 The choice 210:219 The choice of protecting groups in sialyl donors 210:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 5 24 theme Several 782:788 arg1 donors 797:802 Several sialyl donors 782:802 Several sialyl donors studied 782:810 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 5 25 theme disaccharides 848:860 arg1 yields 838:843 yields 838:843 yields of disaccharides 838:860 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 5 26 dep results 829:835 arg1 high 872:875 high 872:875 high 872:875 Several sialyl donors studied showed promising results: yields of disaccharides can be as high as 86% as can be the stereoselectivities (α/β up to 15:1). 28934626 6 27 from changes 1042:1048 arg1 outcome 1057:1063 the outcome 1053:1063 the outcome of sialylation 1053:1078 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 28 theme groups 328:333 arg1 ability 302:308 ability 302:308 ability of different acyl groups to act as hydrogen bond acceptors 302:367 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 29 theme supramolecular 389:402 arg1 structure 404:412 the supramolecular structure 385:412 the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation 385:487 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 0 30 theme Stereoselective 0:14 arg1 sialylation 16:26 Stereoselective sialylation 0:26 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 0:96 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 28934626 6 31 theme O-protecting 983:994 arg1 groups 996:1001 varying acyl O-protecting groups 970:1001 varying acyl O-protecting groups in sialyl donor 970:1017 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 32 theme groups 1292:1297 arg1 properties 1263:1272 electron-withdrawing properties 1242:1272 electron-withdrawing properties of different acyl groups 1242:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 1 33 theme novel 110:114 arg1 donors 123:128 novel sialyl donors 110:128 novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions 110:191 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 4 34 from presence 693:700 arg1 donor 730:734 a sialyl donor 721:734 a sialyl donor 721:734 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 4 35 theme groups 711:716 arg1 presence 693:700 The presence 689:700 The presence of O-TFA groups in a sialyl donor 689:734 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 0 36 theme O-trifluoroacetylated 33:53 arg1 thiosialosides 55:68 O-trifluoroacetylated thiosialosides 33:68 O-trifluoroacetylated thiosialosides 33:68 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 28934626 3 37 theme sialyl 653:658 arg1 donors 660:665 sialyl donors 653:665 sialyl donors without O-TFA groups 653:686 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 6 38 theme properties 1263:1272 arg1 variations 1228:1237 variations 1228:1237 variations of electron-withdrawing properties of different acyl groups 1228:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 1 39 theme sialyl 116:121 arg1 donors 123:128 novel sialyl donors 110:128 novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions 110:191 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 2 40 theme solution 435:442 arg1 mixture 426:432 reaction mixture 417:432 reaction mixture (solution structure) 417:453 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 40 theme solution 435:442 arg1 structure 444:452 solution structure 435:452 solution structure 435:452 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 41 theme acyl 323:326 arg1 groups 328:333 different acyl groups 313:333 different acyl groups 313:333 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 6 42 theme varying 970:976 arg1 groups 996:1001 varying acyl O-protecting groups 970:1001 varying acyl O-protecting groups in sialyl donor 970:1017 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 43 theme substituent 1197:1207 arg1 effects 1209:1215 intramolecular substituent effects 1182:1215 intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups 1182:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 1 44 theme donors 123:128 arg1 series 100:105 A series 98:105 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions 98:191 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 6 45 theme bonding 1170:1176 arg1 influence 1133:1141 the influence 1129:1141 the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups 1129:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 46 theme sialylation 477:487 arg1 outcome 466:472 the outcome 462:472 the outcome of sialylation 462:487 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 46 theme sialylation 477:487 arg1 mixture 426:432 reaction mixture 417:432 reaction mixture (solution structure) 417:453 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 6 47 theme acyl 1287:1290 arg1 groups 1292:1297 different acyl groups 1277:1297 different acyl groups 1277:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 48 theme mixture 426:432 arg1 structure 404:412 the supramolecular structure 385:412 the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation 385:487 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 6 49 theme sialylation 1068:1078 arg1 outcome 1057:1063 the outcome 1053:1063 the outcome of sialylation 1053:1078 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 50 theme sialyl 245:250 arg1 donors 252:257 sialyl donors 245:257 sialyl donors 245:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 51 theme reaction 417:424 arg1 mixture 426:432 reaction mixture 417:432 reaction mixture (solution structure) 417:453 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 51 theme reaction 417:424 arg1 structure 444:452 solution structure 435:452 solution structure 435:452 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 51 theme reaction 417:424 arg1 outcome 466:472 the outcome 462:472 the outcome of sialylation 462:487 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 52 dep hypothesis 272:281 arg1 influence 375:383 influence 375:383 would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation 369:487 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 3 53 theme 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 arg1 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 3 53 theme 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 586:632 arg1 group 577:581 the primary hydroxyl group 556:581 the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose 556:632 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 2 54 theme hydrogen 345:352 arg1 bond 354:357 hydrogen bond 345:357 hydrogen bond acceptors 345:367 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 0 55 theme hydrogen 71:78 arg1 bonding 80:86 hydrogen bonding 71:86 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 0:96 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 28934626 2 56 theme protecting 224:233 arg1 groups 235:240 protecting groups 224:240 protecting groups in sialyl donors 224:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 2 57 theme groups 235:240 arg1 choice 214:219 The choice 210:219 The choice of protecting groups in sialyl donors 210:257 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 0 58 with sialylation 16:26 arg1 thiosialosides 55:68 O-trifluoroacetylated thiosialosides 33:68 O-trifluoroacetylated thiosialosides 33:68 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 28934626 1 59 theme O-trifluoroacetyl 141:157 arg1 groups 165:170 O-trifluoroacetyl (TFA) groups 141:170 O-trifluoroacetyl (TFA) groups 141:170 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 2 60 from variations 288:297 arg1 ability 302:308 ability 302:308 ability of different acyl groups to act as hydrogen bond acceptors 302:367 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 6 61 theme electron-withdrawing 1242:1261 arg1 properties 1263:1272 electron-withdrawing properties 1242:1272 electron-withdrawing properties of different acyl groups 1242:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 62 theme different 1277:1285 arg1 groups 1292:1297 different acyl groups 1277:1297 different acyl groups 1277:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 2 63 theme different 313:321 arg1 groups 328:333 different acyl groups 313:333 different acyl groups 313:333 The choice of protecting groups in sialyl donors was based on hypothesis that variations in ability of different acyl groups to act as hydrogen bond acceptors would influence the supramolecular structure of reaction mixture (solution structure), hence the outcome of sialylation. 28934626 6 64 theme dramatic 1033:1040 arg1 changes 1042:1048 dramatic changes 1033:1048 dramatic changes in the outcome of sialylation 1033:1078 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 65 theme intramolecular 1182:1195 arg1 effects 1209:1215 intramolecular substituent effects 1182:1215 intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups 1182:1297 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 6 66 from groups 996:1001 arg1 donor 1013:1017 sialyl donor 1006:1017 sialyl donor 1006:1017 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 28934626 3 67 theme O-TFA 675:679 arg1 groups 681:686 O-TFA groups 675:686 O-TFA groups 675:686 These glycosyl donors were examined in the model glycosylation of the primary hydroxyl group of 1,2:3,4-di-O-isopropylidene-α-D-galactopyranose in comparison with sialyl donors without O-TFA groups. 28934626 1 68 theme TFA 160:162 arg1 groups 165:170 O-trifluoroacetyl (TFA) groups 141:170 O-trifluoroacetyl (TFA) groups 141:170 A series of novel sialyl donors containing O-trifluoroacetyl (TFA) groups at various positions was synthesized. 28934626 4 69 attach presence 693:700 arg1 donor 730:734 a sialyl donor 721:734 a sialyl donor 721:734 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 4 69 attach presence 693:700 arg2 groups 711:716 O-TFA groups 705:716 O-TFA groups 705:716 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 0 70 dep sialylation 16:26 arg1 bonding 80:86 hydrogen bonding 71:86 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 0:96 Stereoselective sialylation with O-trifluoroacetylated thiosialosides: hydrogen bonding involved? 28934626 4 71 theme sialyl 723:728 arg1 donor 730:734 a sialyl donor 721:734 a sialyl donor 721:734 The presence of O-TFA groups in a sialyl donor strongly affected the outcome of sialylation. 28934626 6 72 theme intermolecular 1146:1159 arg1 bonding 1170:1176 intermolecular hydrogen bonding 1146:1176 intermolecular hydrogen bonding 1146:1176 The results obtained suggest that varying acyl O-protecting groups in sialyl donor may result in dramatic changes in the outcome of sialylation although further studies are required to dissect the influence of intermolecular hydrogen bonding and intramolecular substituent effects related to variations of electron-withdrawing properties of different acyl groups. 26899925 0 0 theme family 96:101 arg1 primers 116:122 glycosyl hydrolase family 28 pectinase primers 77:122 glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi 77:149 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 1 1 from structure 303:311 arg1 polymer 236:242 a prevalent polymer 224:242 a prevalent polymer in plant cell walls that is important in plant defense and structure 224:311 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 1 from structure 303:311 arg1 pectin 216:221 down pectin 211:221 down pectin 211:221 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 1 from structure 303:311 arg1 important 272:280 important 272:280 important 272:280 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 3 2 theme coding 464:469 arg1 genes 471:475 GH28 coding genes 459:475 GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes 459:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 1 3 theme plant 247:251 arg1 walls 258:262 plant cell walls 247:262 plant cell walls 247:262 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 0 4 theme hydrolase 86:94 arg1 primers 116:122 glycosyl hydrolase family 28 pectinase primers 77:122 glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi 77:149 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 7 5 theme GH28 1073:1076 arg1 primers 1078:1084 GH28 primers 1073:1084 GH28 primers 1073:1084 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 1 6 theme wide 175:178 arg1 assortment 180:189 a wide assortment 173:189 a wide assortment of enzymes 173:200 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 7 theme cell 253:256 arg1 walls 258:262 plant cell walls 247:262 plant cell walls 247:262 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 0 8 theme pectinase 106:114 arg1 primers 116:122 glycosyl hydrolase family 28 pectinase primers 77:122 glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi 77:149 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 0 9 from Comparison 0:9 arg1 forests 63:69 temperate forests 53:69 temperate forests 53:69 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 1 10 from defense 291:297 arg1 polymer 236:242 a prevalent polymer 224:242 a prevalent polymer in plant cell walls that is important in plant defense and structure 224:311 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 10 from defense 291:297 arg1 pectin 216:221 down pectin 211:221 down pectin 211:221 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 10 from defense 291:297 arg1 important 272:280 important 272:280 important 272:280 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 7 11 dep % 1191:1191 arg1 39 1189:1190 39 1189:1190 39 1189:1190 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 5 12 from primers 725:731 arg1 PCRs 736:739 PCRs 736:739 PCRs 736:739 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 5 12 from primers 725:731 arg1 DNA 756:758 metagenomic DNA 744:758 metagenomic DNA extracted from senesced tree leaves from different forest ecosystems 744:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 7 13 theme fungal 1159:1164 arg1 communities 1166:1176 fungal communities 1159:1176 fungal communities 1159:1176 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 6 14 dep spacer 989:994 arg1 ITS 997:999 ITS 997:999 ITS 997:999 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 7 15 from Basidiomycetes 1193:1206 arg1 contrast 1120:1127 contrast 1120:1127 contrast 1120:1127 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 3 16 theme GH28 500:503 arg1 sequences 505:513 293 GH28 sequences 496:513 293 GH28 sequences from 40 fungal genomes 496:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 5 17 theme senesced 775:782 arg1 tree 784:787 senesced tree leaves 775:794 senesced tree leaves from different forest ecosystems 775:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 2 18 theme glycosyl 362:369 arg1 family 325:330 One enzyme family 314:330 One enzyme family used to degrade pectin 314:353 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 2 18 theme glycosyl 362:369 arg1 family 381:386 the glycosyl hydrolase family 28 358:389 the glycosyl hydrolase family 28 (GH28) 358:396 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 2 18 theme glycosyl 362:369 arg1 GH28 392:395 GH28 392:395 GH28 392:395 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 4 19 theme Ascomycota 610:619 arg1 cultures 621:628 all Ascomycota cultures 606:628 all Ascomycota cultures tested 606:635 The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. 26899925 9 20 theme ecosystem 1416:1424 arg1 type 1426:1429 ecosystem type 1416:1429 ecosystem type 1416:1429 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 9 21 theme prominent 1361:1369 arg1 role 1371:1374 the prominent role 1357:1374 the prominent role 1357:1374 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 6 22 theme internal 968:975 arg1 spacer 989:994 internal transcribed spacer 968:994 internal transcribed spacer (ITS) amplicon composition using pyrosequencing 968:1042 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 2 23 theme hydrolase 371:379 arg1 family 325:330 One enzyme family 314:330 One enzyme family used to degrade pectin 314:353 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 2 23 theme hydrolase 371:379 arg1 family 381:386 the glycosyl hydrolase family 28 358:389 the glycosyl hydrolase family 28 (GH28) 358:396 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 2 23 theme hydrolase 371:379 arg1 GH28 392:395 GH28 392:395 GH28 392:395 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 9 24 theme possible 1482:1489 arg1 drift 1499:1503 possible genetic drift 1482:1503 possible genetic drift between populations of fungi 1482:1532 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 3 25 from amplification 442:454 arg1 database 484:491 a database 482:491 a database of 293 GH28 sequences from 40 fungal genomes 482:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 7 26 dep 39 1189:1190 arg1 to 1186:1187 to 1186:1187 to 1186:1187 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 1 27 theme enzymes 194:200 arg1 assortment 180:189 a wide assortment 173:189 a wide assortment of enzymes 173:200 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 5 28 theme metagenomic 744:754 arg1 DNA 756:758 metagenomic DNA 744:758 metagenomic DNA extracted from senesced tree leaves from different forest ecosystems 744:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 9 29 theme fungi 1528:1532 arg1 populations 1513:1523 populations 1513:1523 populations of fungi 1513:1532 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 1 30 from important 272:280 arg1 structure 303:311 structure 303:311 structure 303:311 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 30 from important 272:280 arg1 defense 291:297 plant defense 285:297 plant defense 285:297 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 3 31 theme fungal 523:528 arg1 genomes 530:536 40 fungal genomes 520:536 40 fungal genomes 520:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 3 32 from genomes 530:536 arg1 database 484:491 a database 482:491 a database of 293 GH28 sequences from 40 fungal genomes 482:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 3 32 from genomes 530:536 arg1 sequences 505:513 293 GH28 sequences 496:513 293 GH28 sequences from 40 fungal genomes 496:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 6 33 theme amplicon 1002:1009 arg1 composition 1011:1021 internal transcribed spacer (ITS) amplicon composition 968:1021 internal transcribed spacer (ITS) amplicon composition using pyrosequencing 968:1042 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 0 34 theme fungal 31:36 arg1 communities 38:48 pectin-degrading fungal communities 14:48 pectin-degrading fungal communities 14:48 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 8 35 theme forest 1254:1259 arg1 stand 1261:1265 forest stand 1254:1265 forest stand 1254:1265 Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. 26899925 9 36 theme GH28 1390:1393 arg1 composition 1395:1405 GH28 composition 1390:1405 GH28 composition 1390:1405 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 3 37 theme sequences 505:513 arg1 database 484:491 a database 482:491 a database of 293 GH28 sequences from 40 fungal genomes 482:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 4 38 theme GH28 585:588 arg1 pectinases 590:599 GH28 pectinases 585:599 GH28 pectinases from all Ascomycota cultures tested 585:635 The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. 26899925 0 39 theme pectin-degrading 14:29 arg1 communities 38:48 pectin-degrading fungal communities 14:48 pectin-degrading fungal communities 14:48 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 10 40 theme community 1622:1630 arg1 composition 1632:1642 fungal community composition 1615:1642 fungal community composition 1615:1642 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 0 41 theme Ascomycete 134:143 arg1 fungi 145:149 Ascomycete fungi 134:149 Ascomycete fungi 134:149 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 3 42 theme genes 471:475 arg1 amplification 442:454 the amplification 438:454 the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes 438:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 9 43 theme genetic 1491:1497 arg1 drift 1499:1503 possible genetic drift 1482:1503 possible genetic drift between populations of fungi 1482:1532 However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. 26899925 1 44 theme down 211:214 arg1 polymer 236:242 a prevalent polymer 224:242 a prevalent polymer in plant cell walls that is important in plant defense and structure 224:311 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 44 theme down 211:214 arg1 pectin 216:221 down pectin 211:221 down pectin 211:221 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 44 theme down 211:214 arg1 important 272:280 important 272:280 important 272:280 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 45 theme plant 285:289 arg1 defense 291:297 plant defense 285:297 plant defense 285:297 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 10 46 theme ecosystem 1648:1656 arg1 processes 1658:1666 ecosystem processes 1648:1666 ecosystem processes 1648:1666 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 8 47 theme samples 1226:1232 arg1 Analysis 1209:1216 Analysis 1209:1216 Analysis of leaf samples 1209:1232 Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. 26899925 0 48 theme communities 38:48 arg1 Comparison 0:9 Comparison 0:9 Comparison of pectin-degrading fungal communities in temperate forests 0:69 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 6 49 theme GH28 903:906 arg1 genes 908:912 Ascomycota GH28 genes 892:912 Ascomycota GH28 genes 892:912 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 4 50 used used 556:559 arg2 primers 543:549 The primers 539:549 The primers 539:549 The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. 26899925 6 51 theme spacer 989:994 arg1 composition 1011:1021 internal transcribed spacer (ITS) amplicon composition 968:1021 internal transcribed spacer (ITS) amplicon composition using pyrosequencing 968:1042 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 5 52 theme forest 811:816 arg1 ecosystems 818:827 different forest ecosystems 801:827 different forest ecosystems 801:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 1 53 from polymer 236:242 arg1 walls 258:262 plant cell walls 247:262 plant cell walls 247:262 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 8 54 theme fungal 1316:1321 arg1 communities 1323:1333 fungal communities 1316:1333 fungal communities 1316:1333 Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. 26899925 0 55 theme temperate 53:61 arg1 forests 63:69 temperate forests 53:69 temperate forests 53:69 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 4 56 from cultures 621:628 arg1 pectinases 590:599 GH28 pectinases 585:599 GH28 pectinases from all Ascomycota cultures tested 585:635 The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. 26899925 8 57 theme ecosystem 1271:1279 arg1 type 1281:1284 ecosystem type 1271:1284 ecosystem type 1271:1284 Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. 26899925 10 58 contain have 1563:1566 arg1 primers 1550:1556 these primers 1544:1556 these primers 1544:1556 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 10 58 contain have 1563:1566 arg2 utility 1568:1574 utility 1568:1574 utility 1568:1574 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 3 59 theme GH28 459:462 arg1 genes 471:475 GH28 coding genes 459:475 GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes 459:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 5 60 dep tree 784:787 arg1 leaves 789:794 leaves 789:794 leaves 789:794 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 2 61 theme enzyme 318:323 arg1 family 325:330 One enzyme family 314:330 One enzyme family used to degrade pectin 314:353 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 2 61 theme enzyme 318:323 arg1 family 381:386 the glycosyl hydrolase family 28 358:389 the glycosyl hydrolase family 28 (GH28) 358:396 One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). 26899925 7 62 dep Basidiomycetes 1193:1206 arg1 % 1191:1191 % 1191:1191 % 1191:1191 All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communities were up to 39% Basidiomycetes. 26899925 6 63 theme transcribed 977:987 arg1 spacer 989:994 internal transcribed spacer 968:994 internal transcribed spacer (ITS) amplicon composition using pyrosequencing 968:1042 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 10 64 theme fungal 1615:1620 arg1 composition 1632:1642 fungal community composition 1615:1642 fungal community composition 1615:1642 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 4 65 theme Basidiomycota 666:678 arg1 cultures 680:687 three out of seven Basidiomycota cultures 647:687 three out of seven Basidiomycota cultures 647:687 The primers were used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. 26899925 1 66 theme prevalent 226:234 arg1 polymer 236:242 a prevalent polymer 224:242 a prevalent polymer in plant cell walls that is important in plant defense and structure 224:311 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 66 theme prevalent 226:234 arg1 pectin 216:221 down pectin 211:221 down pectin 211:221 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 1 66 theme prevalent 226:234 arg1 important 272:280 important 272:280 important 272:280 Fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. 26899925 0 67 theme glycosyl 77:84 arg1 primers 116:122 glycosyl hydrolase family 28 pectinase primers 77:122 glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi 77:149 Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi. 26899925 8 68 theme leaf 1221:1224 arg1 samples 1226:1232 leaf samples 1221:1232 leaf samples 1221:1232 Analysis of leaf samples indicated that both forest stand and ecosystem type were important in structuring fungal communities. 26899925 5 69 from ecosystems 818:827 arg1 tree 784:787 senesced tree leaves 775:794 senesced tree leaves from different forest ecosystems 775:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 10 70 theme Ascomycetes 1716:1726 arg1 detection 1680:1688 detection 1680:1688 detection of potentially pathogenic Ascomycetes 1680:1726 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 10 70 theme Ascomycetes 1716:1726 arg1 relationships 1593:1605 relationships 1593:1605 relationships between fungal community composition and ecosystem processes 1593:1666 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 5 71 theme different 801:809 arg1 ecosystems 818:827 different forest ecosystems 801:827 different forest ecosystems 801:827 In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senesced tree leaves from different forest ecosystems, followed by cloning and sequencing. 26899925 6 72 theme GH28 938:941 arg1 composition 943:953 GH28 composition 938:953 GH28 composition 938:953 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 6 73 theme Taxonomic 866:874 arg1 specificity 876:886 Taxonomic specificity 866:886 Taxonomic specificity for Ascomycota GH28 genes 866:912 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 26899925 10 74 theme pathogenic 1705:1714 arg1 Ascomycetes 1716:1726 potentially pathogenic Ascomycetes 1693:1726 potentially pathogenic Ascomycetes 1693:1726 Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes. 26899925 3 75 from database 484:491 arg1 genes 471:475 GH28 coding genes 459:475 GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes 459:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 3 75 from database 484:491 arg1 genomes 530:536 40 fungal genomes 520:536 40 fungal genomes 520:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 3 75 from database 484:491 arg1 amplification 442:454 the amplification 438:454 the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes 438:536 In this study we developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from 40 fungal genomes. 26899925 6 76 theme Ascomycota 892:901 arg1 genes 908:912 Ascomycota GH28 genes 892:912 Ascomycota GH28 genes 892:912 Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. 28277614 9 0 theme O-glycan 1802:1809 arg1 composition 1811:1821 this novel O-glycan composition 1791:1821 this novel O-glycan composition 1791:1821 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 6 1 theme novel 1221:1225 arg1 composition 1236:1246 this novel O-glycan composition 1216:1246 this novel O-glycan composition 1216:1246 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 4 2 theme ion 670:672 arg1 mode 674:677 negative ion mode 661:677 negative ion mode 661:677 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 10 3 theme significant 2049:2059 arg1 implications 2061:2072 significant implications 2049:2072 significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system 2049:2197 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 7 4 theme mode 1258:1261 arg1 C18-LC/ESI-MS/MS 1263:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 7 5 theme Positive 1249:1256 arg1 C18-LC/ESI-MS/MS 1263:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 10 6 theme I 2096:2096 arg1 structure 2098:2106 cellobiohydrolase I structure 2078:2106 cellobiohydrolase I structure 2078:2106 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 4 7 theme ion 770:772 arg1 mode 774:777 positive ion mode 761:777 positive ion mode 761:777 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 1 8 theme glycosylation 230:242 arg1 machinery 244:252 eukaryotic glycosylation machinery 219:252 eukaryotic glycosylation machinery 219:252 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 4 9 theme mass 722:725 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 0 10 theme mass 147:150 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 2 11 theme secreted 353:360 arg1 proteins 362:369 secreted proteins 353:369 secreted proteins of RUT-C30 353:380 The N-glycosylation of secreted proteins of RUT-C30 is known to vary depending on culture nutrients but O-glycosylation has been less extensively studied. 28277614 8 12 theme hexuronic 1600:1608 arg1 substituent 1615:1625 the hexuronic acid substituent 1596:1625 the hexuronic acid substituent 1596:1625 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 12 theme hexuronic 1600:1608 arg1 possible 1665:1672 possible 1665:1672 possible 1665:1672 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 10 13 theme recombinant 2148:2158 arg1 proteins 2160:2167 recombinant proteins 2148:2167 recombinant proteins expressed in this host system 2148:2197 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 1 14 theme reesei 262:267 arg1 RUT-C30 269:275 T. reesei RUT-C30 259:275 T. reesei RUT-C30 259:275 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 6 15 theme targeted 1081:1088 arg1 experiments 1094:1104 targeted MS3 experiments 1081:1104 targeted MS3 experiments 1081:1104 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 9 16 theme cellobiohydrolase 1874:1890 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 16 theme cellobiohydrolase 1874:1890 arg1 protein 1922:1928 the most abundant secreted protein 1895:1928 the most abundant secreted protein by T. reesei 1895:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 4 17 theme quadrupole-orbitrap 813:831 arg1 spectrometry 838:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 5 18 theme trap 1048:1051 arg1 fragmentation 1053:1065 standard ion trap fragmentation 1035:1065 standard ion trap fragmentation 1035:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 8 19 theme O-glycan 1630:1637 arg1 structures 1639:1648 O-glycan structures 1630:1648 O-glycan structures 1630:1648 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 1 20 theme RATIONALE 166:174 arg1 production 189:198 RATIONALE High protein production 166:198 RATIONALE High protein production 166:198 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 7 21 theme O-glycans 1443:1451 arg1 I 1413:1413 cellobiohydrolase I 1395:1413 cellobiohydrolase I 1395:1413 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 7 21 theme O-glycans 1443:1451 arg1 carrier 1420:1426 a carrier 1418:1426 a carrier of these novel O-glycans 1418:1451 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 8 22 theme Negative 1462:1469 arg1 trap 1480:1483 Negative mode ion trap 1462:1483 Negative mode ion trap 1462:1483 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 7 23 theme O-glycans 1372:1380 arg1 class 1363:1367 this class 1358:1367 this class of O-glycans 1358:1380 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 5 24 theme detectable 1019:1028 arg1 fragments 1005:1013 glycan fragments 998:1013 glycan fragments not detectable with standard ion trap fragmentation 998:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 9 25 theme secreted 1913:1920 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 25 theme secreted 1913:1920 arg1 protein 1922:1928 the most abundant secreted protein 1895:1928 the most abundant secreted protein by T. reesei 1895:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 10 26 theme proteins 2160:2167 arg1 activity 2136:2143 the activity 2132:2143 the activity of recombinant proteins expressed in this host system 2132:2197 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 10 26 theme proteins 2160:2167 arg1 activity 2112:2119 activity 2112:2119 activity 2112:2119 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 10 26 theme proteins 2160:2167 arg1 structure 2098:2106 cellobiohydrolase I structure 2078:2106 cellobiohydrolase I structure 2078:2106 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 4 27 theme ionisation 795:804 arg1 spectrometry 838:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 0 28 theme ion 93:95 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 5 29 theme standard 1035:1042 arg1 fragmentation 1053:1065 standard ion trap fragmentation 1035:1065 standard ion trap fragmentation 1035:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 8 30 theme higher 1485:1490 arg1 dissociation 1517:1528 Negative mode ion trap higher energy collision-induced dissociation 1462:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation 1454:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 0 31 theme higher 102:107 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 8 32 theme ion 1679:1681 arg1 CID 1688:1690 ion trap CID 1679:1690 ion trap CID 1679:1690 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 33 theme collision-induced 1499:1515 arg1 dissociation 1517:1528 Negative mode ion trap higher energy collision-induced dissociation 1462:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation 1454:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 0 34 theme collision-induced 116:132 arg1 dissociation 134:145 energy collision-induced dissociation 109:145 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 5 35 theme higher 936:941 arg1 tHCD 982:985 tHCD 982:985 tHCD 982:985 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 35 theme higher 936:941 arg1 dissociation 968:979 higher energy collision-induced dissociation 936:979 higher energy collision-induced dissociation (tHCD) 936:986 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 6 36 theme trap 1195:1198 arg1 CID 1200:1202 ion trap CID 1191:1202 ion trap CID 1191:1202 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 6 37 contain containing 1136:1145 arg1 structures 1125:1134 structures 1125:1134 structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition 1125:1246 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 6 37 contain containing 1136:1145 arg2 acid 1157:1160 hexuronic acid 1147:1160 hexuronic acid 1147:1160 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 5 38 theme collision-induced 950:966 arg1 tHCD 982:985 tHCD 982:985 tHCD 982:985 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 38 theme collision-induced 950:966 arg1 dissociation 968:979 higher energy collision-induced dissociation 936:979 higher energy collision-induced dissociation (tHCD) 936:986 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 8 39 theme novel 1708:1712 arg1 composition 1723:1733 the novel O-glycan composition 1704:1733 the novel O-glycan composition to include hexuronic acid 1704:1759 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 40 theme MS3 1561:1563 arg1 experiments 1565:1575 targeted MS3 experiments 1552:1575 targeted MS3 experiments 1552:1575 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 0 41 theme glucuronidated 41:54 arg1 glycans 65:71 glucuronidated O-linked glycans 41:71 glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry 41:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 7 42 theme novel 1437:1441 arg1 O-glycans 1443:1451 these novel O-glycans 1431:1451 these novel O-glycans 1431:1451 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 0 43 theme negative 79:86 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 3 44 theme porous 562:567 arg1 carbon 581:586 porous graphitised carbon and C-18 liquid chromatography 562:617 carbon 581:586 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 5 45 theme ion 927:929 arg1 trap 931:934 ion trap 927:934 ion trap 927:934 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 6 46 with possible 1177:1184 arg1 CID 1200:1202 ion trap CID 1191:1202 ion trap CID 1191:1202 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 8 47 dep RESULTS 1454:1460 arg1 dissociation 1517:1528 Negative mode ion trap higher energy collision-induced dissociation 1462:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation 1454:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 5 48 gly glycopeptides 903:915 arg2 trap 931:934 ion trap 927:934 ion trap 927:934 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 48 gly glycopeptides 903:915 arg2 glycopeptides 903:915 glycopeptides 903:915 glycopeptides including ion trap 903:934 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 1 49 theme protein 181:187 arg1 production 189:198 RATIONALE High protein production 166:198 RATIONALE High protein production 166:198 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 4 50 theme electrospray 682:693 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 11 51 dep & 2228:2228 arg1 Sons 2230:2233 Sons 2230:2233 Sons 2230:2233 Copyright © 2017 John Wiley & Sons, Ltd. 28277614 9 52 attach present 1839:1845 arg1 domain 1864:1869 the catalytic domain 1850:1869 the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei 1850:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 52 attach present 1839:1845 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 52 attach present 1839:1845 arg2 composition 1811:1821 this novel O-glycan composition 1791:1821 this novel O-glycan composition 1791:1821 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 3 53 theme C-18 592:595 arg1 chromatography 604:617 porous graphitised carbon and C-18 liquid chromatography 562:617 chromatography 604:617 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 1 54 theme suitable 279:286 arg1 host 299:302 a suitable expression host 277:302 a suitable expression host for recombinant proteins 277:327 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 9 55 theme novel 1796:1800 arg1 composition 1811:1821 this novel O-glycan composition 1791:1821 this novel O-glycan composition 1791:1821 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 6 56 theme O-glycan 1227:1234 arg1 composition 1236:1246 this novel O-glycan composition 1216:1246 this novel O-glycan composition 1216:1246 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 4 57 theme linear 706:711 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 4 58 gly glycopeptides 744:756 arg2 glycopeptides 744:756 glycopeptides 744:756 glycopeptides 744:756 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 10 59 from sugars 2013:2018 arg1 fungi 2023:2027 fungi 2023:2027 fungi 2023:2027 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 4 60 theme negative 661:668 arg1 mode 674:677 negative ion mode 661:677 negative ion mode 661:677 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 2 61 gly N-glycosylation 334:348 arg1 proteins 362:369 secreted proteins 353:369 secreted proteins of RUT-C30 353:380 The N-glycosylation of secreted proteins of RUT-C30 is known to vary depending on culture nutrients but O-glycosylation has been less extensively studied. 28277614 1 62 theme recombinant 308:318 arg1 proteins 320:327 recombinant proteins 308:327 recombinant proteins 308:327 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 8 63 theme ion 1476:1478 arg1 trap 1480:1483 Negative mode ion trap 1462:1483 Negative mode ion trap 1462:1483 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 10 64 contain have 2044:2047 arg1 they 2033:2036 they 2033:2036 they 2033:2036 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 10 64 contain have 2044:2047 arg2 implications 2061:2072 significant implications 2049:2072 significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system 2049:2197 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 5 65 theme glycan 998:1003 arg1 fragments 1005:1013 glycan fragments 998:1013 glycan fragments not detectable with standard ion trap fragmentation 998:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 0 66 theme glycans 65:71 arg1 characterisation 21:36 structural characterisation 10:36 structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry 10:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 10 67 theme cellobiohydrolase 2078:2094 arg1 I 2096:2096 cellobiohydrolase I 2078:2096 cellobiohydrolase I structure 2078:2106 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 2 68 theme RUT-C30 374:380 arg1 proteins 362:369 secreted proteins 353:369 secreted proteins of RUT-C30 353:380 The N-glycosylation of secreted proteins of RUT-C30 is known to vary depending on culture nutrients but O-glycosylation has been less extensively studied. 28277614 1 69 theme eukaryotic 219:228 arg1 machinery 244:252 eukaryotic glycosylation machinery 219:252 eukaryotic glycosylation machinery 219:252 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 0 70 link O-linked 56:63 arg1 glycans 65:71 glucuronidated O-linked glycans 41:71 glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry 41:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 4 71 theme positive 761:768 arg1 mode 774:777 positive ion mode 761:777 positive ion mode 761:777 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 9 72 theme glycopeptide 1768:1779 arg1 analysis 1781:1788 glycopeptide analysis 1768:1788 glycopeptide analysis 1768:1788 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 2 73 theme proteins 362:369 arg1 N-glycosylation 334:348 The N-glycosylation 330:348 The N-glycosylation of secreted proteins of RUT-C30 330:380 The N-glycosylation of secreted proteins of RUT-C30 is known to vary depending on culture nutrients but O-glycosylation has been less extensively studied. 28277614 5 74 dep O-glycans 889:897 arg1 tHCD 982:985 tHCD 982:985 tHCD 982:985 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 74 dep O-glycans 889:897 arg1 dissociation 968:979 higher energy collision-induced dissociation 936:979 higher energy collision-induced dissociation (tHCD) 936:986 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 8 75 theme acid 1610:1613 arg1 substituent 1615:1625 the hexuronic acid substituent 1596:1625 the hexuronic acid substituent 1596:1625 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 75 theme acid 1610:1613 arg1 possible 1665:1672 possible 1665:1672 possible 1665:1672 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 76 theme mode 1471:1474 arg1 trap 1480:1483 Negative mode ion trap 1462:1483 Negative mode ion trap 1462:1483 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 4 77 theme trap 717:720 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 1 78 theme T. 259:260 arg1 RUT-C30 269:275 T. reesei RUT-C30 259:275 T. reesei RUT-C30 259:275 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 6 79 theme MS3 1090:1092 arg1 experiments 1094:1104 targeted MS3 experiments 1081:1104 targeted MS3 experiments 1081:1104 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 5 80 theme ion 1044:1046 arg1 fragmentation 1053:1065 standard ion trap fragmentation 1035:1065 standard ion trap fragmentation 1035:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 9 81 theme I 1892:1892 arg1 domain 1864:1869 the catalytic domain 1850:1869 the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei 1850:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 81 theme I 1892:1892 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 8 82 theme structures 1639:1648 arg1 substituent 1615:1625 the hexuronic acid substituent 1596:1625 the hexuronic acid substituent 1596:1625 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 8 82 theme structures 1639:1648 arg1 possible 1665:1672 possible 1665:1672 possible 1665:1672 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 4 83 theme hybrid 806:811 arg1 spectrometry 838:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 3 84 gly glycopeptides 507:519 arg1 proteins 535:542 secreted proteins 526:542 secreted proteins 526:542 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 3 84 gly glycopeptides 507:519 arg2 glycopeptides 507:519 glycopeptides 507:519 glycopeptides 507:519 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 4 85 theme mass 833:836 arg1 spectrometry 838:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 0 86 theme mode 88:91 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 10 87 theme reported 1976:1983 arg1 CONCLUSIONS 1944:1954 CONCLUSIONS 1944:1954 CONCLUSIONS These 1944:1960 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 10 87 theme reported 1976:1983 arg1 O-glycans 1985:1993 the first reported O-glycans 1966:1993 the first reported O-glycans to contain acidic sugars in fungi 1966:2027 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 1 88 theme High 176:179 arg1 production 189:198 RATIONALE High protein production 166:198 RATIONALE High protein production 166:198 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 9 89 theme abundant 1904:1911 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 89 theme abundant 1904:1911 arg1 protein 1922:1928 the most abundant secreted protein 1895:1928 the most abundant secreted protein by T. reesei 1895:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 5 90 theme Tandem 852:857 arg1 spectrometry 864:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 0 91 theme trap 97:100 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 1 92 with production 189:198 arg1 machinery 244:252 eukaryotic glycosylation machinery 219:252 eukaryotic glycosylation machinery 219:252 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 6 93 theme hexuronic 1147:1155 arg1 acid 1157:1160 hexuronic acid 1147:1160 hexuronic acid 1147:1160 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 9 94 theme catalytic 1854:1862 arg1 domain 1864:1869 the catalytic domain 1850:1869 the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei 1850:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 94 theme catalytic 1854:1862 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 4 95 theme electrospray 782:793 arg1 spectrometry 838:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry 782:849 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 0 96 theme energy 109:114 arg1 dissociation 134:145 energy collision-induced dissociation 109:145 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 8 97 theme trap 1683:1686 arg1 CID 1688:1690 ion trap CID 1679:1690 ion trap CID 1679:1690 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 5 98 with detectable 1019:1028 arg1 fragmentation 1053:1065 standard ion trap fragmentation 1035:1065 standard ion trap fragmentation 1035:1065 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 10 99 theme host 2187:2190 arg1 system 2192:2197 this host system 2182:2197 this host system 2182:2197 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 0 100 theme dissociation 134:145 arg1 spectrometry 152:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 negative mode ion trap higher energy collision-induced dissociation mass spectrometry 79:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 10 101 dep CONCLUSIONS 1944:1954 arg1 These 1956:1960 These 1956:1960 These 1956:1960 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 8 102 theme energy 1492:1497 arg1 dissociation 1517:1528 Negative mode ion trap higher energy collision-induced dissociation 1462:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation 1454:1528 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 5 103 theme mass 859:862 arg1 spectrometry 864:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 0 104 theme structural 10:19 arg1 characterisation 21:36 structural characterisation 10:36 structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry 10:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 5 105 used used 881:884 arg2 spectrometry 864:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry 852:875 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 106 theme energy 943:948 arg1 tHCD 982:985 tHCD 982:985 tHCD 982:985 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 5 106 theme energy 943:948 arg1 dissociation 968:979 higher energy collision-induced dissociation 936:979 higher energy collision-induced dissociation (tHCD) 936:986 Tandem mass spectrometry was used on O-glycans and glycopeptides including ion trap higher energy collision-induced dissociation (tHCD) to detect glycan fragments not detectable with standard ion trap fragmentation. 28277614 6 107 theme ion 1191:1193 arg1 CID 1200:1202 ion trap CID 1191:1202 ion trap CID 1191:1202 tHCD allowed targeted MS3 experiments to be performed on structures containing hexuronic acid, which was not possible with ion trap CID, validating this novel O-glycan composition. 28277614 8 108 theme O-glycan 1714:1721 arg1 composition 1723:1733 the novel O-glycan composition 1704:1733 the novel O-glycan composition to include hexuronic acid 1704:1759 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 9 109 gly glycopeptide 1768:1779 arg2 glycopeptide 1768:1779 glycopeptide analysis 1768:1788 glycopeptide analysis 1768:1788 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 0 110 theme O-linked 56:63 arg1 glycans 65:71 glucuronidated O-linked glycans 41:71 glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry 41:163 Enhancing structural characterisation of glucuronidated O-linked glycans using negative mode ion trap higher energy collision-induced dissociation mass spectrometry. 28277614 3 111 dep METHODS 485:491 arg1 O-Glycans 493:501 O-Glycans 493:501 O-Glycans 493:501 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 10 112 theme acidic 2006:2011 arg1 sugars 2013:2018 acidic sugars 2006:2018 acidic sugars in fungi 2006:2027 CONCLUSIONS These are the first reported O-glycans to contain acidic sugars in fungi and they could have significant implications for cellobiohydrolase I structure and activity as well as the activity of recombinant proteins expressed in this host system. 28277614 8 113 theme targeted 1552:1559 arg1 experiments 1565:1575 targeted MS3 experiments 1552:1575 targeted MS3 experiments 1552:1575 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 3 114 theme secreted 526:533 arg1 proteins 535:542 secreted proteins 526:542 secreted proteins 526:542 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 7 115 used used 1284:1287 arg2 C18-LC/ESI-MS/MS 1263:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS 1249:1278 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 8 116 theme hexuronic 1746:1754 arg1 acid 1756:1759 hexuronic acid 1746:1759 hexuronic acid 1746:1759 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 3 117 from proteins 535:542 arg1 METHODS 485:491 METHODS 485:491 METHODS 485:491 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 3 117 from proteins 535:542 arg1 glycopeptides 507:519 glycopeptides 507:519 glycopeptides 507:519 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 3 118 theme graphitised 569:579 arg1 carbon 581:586 porous graphitised carbon and C-18 liquid chromatography 562:617 carbon 581:586 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 1 119 with secretion 204:212 arg1 machinery 244:252 eukaryotic glycosylation machinery 219:252 eukaryotic glycosylation machinery 219:252 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 28277614 4 120 theme ionisation 695:704 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 7 121 gly glycopeptides 1318:1330 arg2 glycopeptides 1318:1330 glycopeptides 1318:1330 glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans 1318:1451 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 2 122 theme culture 412:418 arg1 nutrients 420:428 culture nutrients 412:428 culture nutrients 412:428 The N-glycosylation of secreted proteins of RUT-C30 is known to vary depending on culture nutrients but O-glycosylation has been less extensively studied. 28277614 9 123 located present 1839:1845 arg1 domain 1864:1869 the catalytic domain 1850:1869 the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei 1850:1941 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 123 located present 1839:1845 arg1 I 1892:1892 cellobiohydrolase I 1874:1892 cellobiohydrolase I 1874:1892 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 9 123 located present 1839:1845 arg2 composition 1811:1821 this novel O-glycan composition 1791:1821 this novel O-glycan composition 1791:1821 Using glycopeptide analysis, this novel O-glycan composition was found to be present on the catalytic domain of cellobiohydrolase I, the most abundant secreted protein by T. reesei. 28277614 7 124 theme cellobiohydrolase 1395:1411 arg1 I 1413:1413 cellobiohydrolase I 1395:1413 cellobiohydrolase I 1395:1413 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 7 124 theme cellobiohydrolase 1395:1411 arg1 carrier 1420:1426 a carrier 1418:1426 a carrier of these novel O-glycans 1418:1451 Positive mode C18-LC/ESI-MS/MS was used to identify and characterise glycopeptides found to be modified with this class of O-glycans, identifying cellobiohydrolase I as a carrier of these novel O-glycans. 28277614 8 125 with possible 1665:1672 arg1 CID 1688:1690 ion trap CID 1679:1690 ion trap CID 1679:1690 RESULTS Negative mode ion trap higher energy collision-induced dissociation allowed detection and targeted MS3 experiments to be performed on the hexuronic acid substituent of O-glycan structures, which was not possible with ion trap CID, validating the novel O-glycan composition to include hexuronic acid. 28277614 4 126 theme ion 713:715 arg1 spectrometry 727:738 electrospray ionisation linear ion trap mass spectrometry 682:738 electrospray ionisation linear ion trap mass spectrometry 682:738 O-Glycans were analysed in negative ion mode by electrospray ionisation linear ion trap mass spectrometry and glycopeptides in positive ion mode by electrospray ionisation hybrid quadrupole-orbitrap mass spectrometry. 28277614 3 127 theme liquid 597:602 arg1 chromatography 604:617 porous graphitised carbon and C-18 liquid chromatography 562:617 chromatography 604:617 METHODS O-Glycans and glycopeptides from secreted proteins were separated by porous graphitised carbon and C-18 liquid chromatography, respectively. 28277614 1 128 theme expression 288:297 arg1 host 299:302 a suitable expression host 277:302 a suitable expression host for recombinant proteins 277:327 RATIONALE High protein production and secretion with eukaryotic glycosylation machinery make T. reesei RUT-C30 a suitable expression host for recombinant proteins. 26322380 10 0 theme targeting 1728:1736 arg1 fields 1704:1709 the fields 1700:1709 the fields of imaging, drug targeting, and biomarkers for lung cancer 1700:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 0 1 theme Differential 0:11 arg1 Patterns 29:36 Differential N-Glycosylation Patterns 0:36 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue 0:66 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue. 26322380 8 2 from tissue 1394:1399 arg1 data 1364:1367 publicly available gene expression data 1329:1367 publicly available gene expression data from lung adenocarcinoma tissue 1329:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 4 3 theme tumor 667:671 arg1 tissue 673:678 lung tumor tissue 662:678 lung tumor tissue 662:678 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 4 4 gly glycosylation 636:648 arg1 tissue 673:678 lung tumor tissue 662:678 lung tumor tissue 662:678 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 4 4 gly glycosylation 636:648 arg1 tissue 697:702 nonmalignant tissue 684:702 nonmalignant tissue 684:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 5 5 theme glycan 872:877 arg1 structures 879:888 29 differentially expressed (FDR < 0.05) glycan structures 831:888 29 differentially expressed (FDR < 0.05) glycan structures 831:888 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 6 6 theme tumor 975:979 arg1 tissue 981:986 tumor tissue 975:986 tumor tissue 975:986 The levels of several oligomannose type glycans were upregulated in tumor tissue. 26322380 4 7 theme individual 720:729 arg1 structures 738:747 individual glycan structures 720:747 individual glycan structures using nLC-chip-TOF-MS 720:769 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 7 8 theme hybrid 1068:1073 arg1 type 1075:1078 the hybrid type 1064:1078 the hybrid type 1064:1078 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 3 9 theme biological 543:552 arg1 mechanisms 554:563 the underlying biological mechanisms 528:563 the underlying biological mechanisms of aberrant glycosylation in lung cancer 528:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 5 10 theme expressed 849:857 arg1 structures 879:888 29 differentially expressed (FDR < 0.05) glycan structures 831:888 29 differentially expressed (FDR < 0.05) glycan structures 831:888 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 4 11 theme glycosylation 636:648 arg1 patterns 650:657 differential glycosylation patterns 623:657 differential glycosylation patterns of lung tumor tissue and nonmalignant tissue 623:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 3 12 theme lung 594:597 arg1 cancer 599:604 lung cancer 594:604 lung cancer 594:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 7 13 theme galactosylated 1018:1031 arg1 glycans 1033:1039 fully galactosylated glycans 1012:1039 fully galactosylated glycans 1012:1039 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 5 14 theme FDR 860:862 arg1 < 864:864 FDR < 0.05 860:869 FDR < 0.05 860:869 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 6 15 theme glycans 947:953 arg1 levels 911:916 The levels 907:916 The levels of several oligomannose type glycans 907:953 The levels of several oligomannose type glycans were upregulated in tumor tissue. 26322380 10 16 used used 1612:1615 arg2 results 1596:1602 These results 1590:1602 These results 1590:1602 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 10 16 used used 1612:1615 arg2 point 1631:1635 a starting point 1620:1635 a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1620:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 7 17 with glycans 1188:1194 arg1 fucose 1208:1213 fucose 1208:1213 fucose 1208:1213 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 8 18 theme glycosylation 1279:1291 arg1 regulation 1253:1262 the regulation 1249:1262 the regulation of the altered glycosylation 1249:1291 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 3 19 theme glycosylation 577:589 arg1 mechanisms 554:563 the underlying biological mechanisms 528:563 the underlying biological mechanisms of aberrant glycosylation in lung cancer 528:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 9 20 theme observed 1496:1503 arg1 changes 1514:1520 the observed N-glycan changes 1492:1520 the observed N-glycan changes 1492:1520 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 6 21 theme oligomannose 929:940 arg1 glycans 947:953 several oligomannose type glycans 921:953 several oligomannose type glycans 921:953 The levels of several oligomannose type glycans were upregulated in tumor tissue. 26322380 1 22 theme diagnostic 132:141 arg1 tools 143:147 better screening and diagnostic tools 111:147 tools 143:147 To decrease the mortality of lung cancer, better screening and diagnostic tools as well as treatment options are needed. 26322380 2 23 theme post-translational 232:249 arg1 modifications 251:263 the major post-translational modifications 222:263 the major post-translational modifications that is altered in cancer 222:289 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 0 24 from Patterns 29:36 arg1 Tissue 61:66 Lung Adenocarcinoma Tissue 41:66 Lung Adenocarcinoma Tissue 41:66 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue. 26322380 4 25 theme tissue 697:702 arg1 patterns 650:657 differential glycosylation patterns 623:657 differential glycosylation patterns of lung tumor tissue and nonmalignant tissue 623:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 5 26 theme tissue 778:783 arg1 samples 785:791 tissue samples 778:791 tissue samples from 42 lung adenocarcinoma patients 778:828 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 9 27 theme expressed 1558:1566 arg1 glycosyltransferases 1568:1587 differentially expressed glycosyltransferases 1543:1587 differentially expressed glycosyltransferases 1543:1587 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 8 28 theme gene 1348:1351 arg1 data 1364:1367 publicly available gene expression data 1329:1367 publicly available gene expression data from lung adenocarcinoma tissue 1329:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 10 29 theme glycan 1677:1682 arg1 applications 1684:1695 clinical glycan applications 1668:1695 clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1668:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 7 30 theme low-galactosylated 1169:1186 arg1 glycans 1188:1194 non- or low-galactosylated glycans 1161:1194 non- or low-galactosylated glycans mostly with fucose 1161:1213 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 4 31 theme glycan 731:736 arg1 structures 738:747 individual glycan structures 720:747 individual glycan structures using nLC-chip-TOF-MS 720:769 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 7 32 theme non- 1161:1164 arg1 glycans 1188:1194 non- or low-galactosylated glycans 1161:1194 non- or low-galactosylated glycans mostly with fucose 1161:1213 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 1 33 theme lung 98:101 arg1 cancer 103:108 lung cancer 98:108 lung cancer 98:108 To decrease the mortality of lung cancer, better screening and diagnostic tools as well as treatment options are needed. 26322380 8 34 theme lung 1374:1377 arg1 tissue 1394:1399 lung adenocarcinoma tissue 1374:1399 lung adenocarcinoma tissue 1374:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 3 35 from glycosylation 577:589 arg1 cancer 599:604 lung cancer 594:604 lung cancer 594:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 10 36 theme lung 1758:1761 arg1 cancer 1763:1768 lung cancer 1758:1768 lung cancer 1758:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 3 37 theme structures 395:404 arg1 understanding 367:379 A better understanding 358:379 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue 358:459 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 3 37 theme structures 395:404 arg1 desirable 471:479 desirable 471:479 desirable 471:479 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 3 38 theme lung 443:446 arg1 tissue 454:459 lung tumor tissue 443:459 lung tumor tissue 443:459 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 0 39 theme Lung 41:44 arg1 Tissue 61:66 Lung Adenocarcinoma Tissue 41:66 Lung Adenocarcinoma Tissue 41:66 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue. 26322380 2 40 theme glycan 326:331 arg1 structures 333:342 glycan structures 326:342 glycan structures 326:342 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 5 41 theme lung 801:804 arg1 patients 821:828 42 lung adenocarcinoma patients 798:828 42 lung adenocarcinoma patients 798:828 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 8 42 theme lung 1426:1429 arg1 tissue 1431:1436 nonmalignant lung tissue 1413:1436 nonmalignant lung tissue 1413:1436 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 10 43 theme imaging 1714:1720 arg1 fields 1704:1709 the fields 1700:1709 the fields of imaging, drug targeting, and biomarkers for lung cancer 1700:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 4 44 theme tissue 673:678 arg1 patterns 650:657 differential glycosylation patterns 623:657 differential glycosylation patterns of lung tumor tissue and nonmalignant tissue 623:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 3 45 theme tumor 448:452 arg1 tissue 454:459 lung tumor tissue 443:459 lung tumor tissue 443:459 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 10 46 theme biomarkers 1743:1752 arg1 fields 1704:1709 the fields 1700:1709 the fields of imaging, drug targeting, and biomarkers for lung cancer 1700:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 6 47 theme several 921:927 arg1 glycans 947:953 several oligomannose type glycans 921:953 several oligomannose type glycans 921:953 The levels of several oligomannose type glycans were upregulated in tumor tissue. 26322380 10 48 from fields 1704:1709 arg1 development 1653:1663 the further development 1641:1663 the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1641:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 9 49 from origin 1533:1538 arg1 glycosyltransferases 1568:1587 differentially expressed glycosyltransferases 1543:1587 differentially expressed glycosyltransferases 1543:1587 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 10 50 theme drug 1723:1726 arg1 targeting 1728:1736 drug targeting 1723:1736 drug targeting 1723:1736 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 3 51 theme greater 507:513 arg1 insight 515:521 greater insight 507:521 greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer 507:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 1 52 theme treatment 160:168 arg1 options 170:176 treatment options 160:176 better screening and diagnostic tools as well as treatment options 111:176 To decrease the mortality of lung cancer, better screening and diagnostic tools as well as treatment options are needed. 26322380 7 53 theme cancerous 1125:1133 arg1 tissue 1135:1140 cancerous tissue 1125:1140 cancerous tissue 1125:1140 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 10 54 theme starting 1622:1629 arg1 results 1596:1602 These results 1590:1602 These results 1590:1602 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 10 54 theme starting 1622:1629 arg1 point 1631:1635 a starting point 1620:1635 a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1620:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 4 55 theme lung 662:665 arg1 tissue 673:678 lung tumor tissue 662:678 lung tumor tissue 662:678 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 3 56 theme underlying 532:541 arg1 mechanisms 554:563 the underlying biological mechanisms 528:563 the underlying biological mechanisms of aberrant glycosylation in lung cancer 528:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 10 57 theme further 1645:1651 arg1 development 1653:1663 the further development 1641:1663 the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1641:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 2 58 theme major 226:230 arg1 modifications 251:263 the major post-translational modifications 222:263 the major post-translational modifications that is altered in cancer 222:289 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 4 59 theme differential 623:634 arg1 patterns 650:657 differential glycosylation patterns 623:657 differential glycosylation patterns of lung tumor tissue and nonmalignant tissue 623:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 3 60 theme aberrant 568:575 arg1 glycosylation 577:589 aberrant glycosylation 568:589 aberrant glycosylation in lung cancer 568:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 7 61 theme glycans 1033:1039 arg1 levels 1002:1007 levels 1002:1007 levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, 1002:1105 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 1 62 theme better 111:116 arg1 screening 118:126 better screening and diagnostic tools 111:147 screening 118:126 To decrease the mortality of lung cancer, better screening and diagnostic tools as well as treatment options are needed. 26322380 6 63 theme type 942:945 arg1 glycans 947:953 several oligomannose type glycans 921:953 several oligomannose type glycans 921:953 The levels of several oligomannose type glycans were upregulated in tumor tissue. 26322380 8 64 theme altered 1271:1277 arg1 glycosylation 1279:1291 the altered glycosylation 1267:1291 the altered glycosylation 1267:1291 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 9 65 theme N-glycan 1505:1512 arg1 changes 1514:1520 the observed N-glycan changes 1492:1520 the observed N-glycan changes 1492:1520 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 3 66 from mechanisms 554:563 arg1 cancer 599:604 lung cancer 594:604 lung cancer 594:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 2 67 theme modifications 251:263 arg1 modifications 251:263 the major post-translational modifications 222:263 the major post-translational modifications that is altered in cancer 222:289 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 2 67 theme modifications 251:263 arg1 one 215:217 one 215:217 one 215:217 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 7 68 dep decreased 1112:1120 arg1 whereas 1143:1149 whereas 1143:1149 whereas 1143:1149 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 8 69 theme glycomics 1298:1306 arg1 data 1308:1311 the glycomics data 1294:1311 the glycomics data 1294:1311 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 4 70 theme nonmalignant 684:695 arg1 tissue 697:702 nonmalignant tissue 684:702 nonmalignant tissue 684:702 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 7 71 theme glycans 1188:1194 arg1 levels 1151:1156 levels 1151:1156 levels of non- or low-galactosylated glycans mostly with fucose 1151:1213 Furthermore, levels of fully galactosylated glycans, some of which were of the hybrid type and mostly without fucose, were decreased in cancerous tissue, whereas levels of non- or low-galactosylated glycans mostly with fucose were increased. 26322380 3 72 from cancer 599:604 arg1 mechanisms 554:563 the underlying biological mechanisms 528:563 the underlying biological mechanisms of aberrant glycosylation in lung cancer 528:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 8 73 theme available 1338:1346 arg1 data 1364:1367 publicly available gene expression data 1329:1367 publicly available gene expression data from lung adenocarcinoma tissue 1329:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 10 74 theme clinical 1668:1675 arg1 applications 1684:1695 clinical glycan applications 1668:1695 clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1668:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 5 75 theme adenocarcinoma 806:819 arg1 patients 821:828 42 lung adenocarcinoma patients 798:828 42 lung adenocarcinoma patients 798:828 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 5 76 from patients 821:828 arg1 samples 785:791 tissue samples 778:791 tissue samples from 42 lung adenocarcinoma patients 778:828 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 8 77 theme expression 1353:1362 arg1 data 1364:1367 publicly available gene expression data 1329:1367 publicly available gene expression data from lung adenocarcinoma tissue 1329:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 1 78 theme cancer 103:108 arg1 mortality 85:93 the mortality 81:93 the mortality of lung cancer 81:108 To decrease the mortality of lung cancer, better screening and diagnostic tools as well as treatment options are needed. 26322380 3 79 theme better 360:365 arg1 understanding 367:379 A better understanding 358:379 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue 358:459 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 3 79 theme better 360:365 arg1 desirable 471:479 desirable 471:479 desirable 471:479 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 4 80 theme structures 738:747 arg1 level 711:715 the level 707:715 the level of individual glycan structures using nLC-chip-TOF-MS 707:769 Here, we assess differential glycosylation patterns of lung tumor tissue and nonmalignant tissue at the level of individual glycan structures using nLC-chip-TOF-MS. 26322380 0 81 theme Adenocarcinoma 46:59 arg1 Tissue 61:66 Lung Adenocarcinoma Tissue 41:66 Lung Adenocarcinoma Tissue 41:66 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue. 26322380 3 82 gly glycosylation 577:589 arg1 cancer 599:604 lung cancer 594:604 lung cancer 594:604 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 8 83 theme adenocarcinoma 1379:1392 arg1 tissue 1394:1399 lung adenocarcinoma tissue 1374:1399 lung adenocarcinoma tissue 1374:1399 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 10 84 theme applications 1684:1695 arg1 development 1653:1663 the further development 1641:1663 the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer 1641:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 10 85 from development 1653:1663 arg1 fields 1704:1709 the fields 1700:1709 the fields of imaging, drug targeting, and biomarkers for lung cancer 1700:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 3 86 theme glycan 388:393 arg1 structures 395:404 the glycan structures 384:404 the glycan structures that are differentially regulated in lung tumor tissue 384:459 A better understanding of the glycan structures that are differentially regulated in lung tumor tissue is highly desirable and will allow us to gain greater insight into the underlying biological mechanisms of aberrant glycosylation in lung cancer. 26322380 9 87 with consistent 1455:1464 arg1 possibility 1475:1485 the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases 1471:1587 the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases 1471:1587 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 9 88 contain have 1522:1525 arg1 changes 1514:1520 the observed N-glycan changes 1492:1520 the observed N-glycan changes 1492:1520 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 9 88 contain have 1522:1525 arg2 origin 1533:1538 their origin 1527:1538 their origin in differentially expressed glycosyltransferases 1527:1587 The results are consistent with the possibility that the observed N-glycan changes have their origin in differentially expressed glycosyltransferases. 26322380 2 89 theme Protein 190:196 arg1 glycosylation 198:210 Protein glycosylation 190:210 Protein glycosylation 190:210 Protein glycosylation is one of the major post-translational modifications that is altered in cancer, but it is not exactly clear which glycan structures are affected. 26322380 8 90 theme nonmalignant 1413:1424 arg1 tissue 1431:1436 nonmalignant lung tissue 1413:1436 nonmalignant lung tissue 1413:1436 To further assess the regulation of the altered glycosylation, the glycomics data was compared to publicly available gene expression data from lung adenocarcinoma tissue compared to nonmalignant lung tissue. 26322380 10 91 from applications 1684:1695 arg1 fields 1704:1709 the fields 1700:1709 the fields of imaging, drug targeting, and biomarkers for lung cancer 1700:1768 These results will be used as a starting point for the further development of clinical glycan applications in the fields of imaging, drug targeting, and biomarkers for lung cancer. 26322380 5 92 dep expressed 849:857 arg1 < 864:864 FDR < 0.05 860:869 FDR < 0.05 860:869 Using tissue samples from 42 lung adenocarcinoma patients, 29 differentially expressed (FDR < 0.05) glycan structures were identified. 26322380 0 93 theme N-Glycosylation 13:27 arg1 Patterns 29:36 Differential N-Glycosylation Patterns 0:36 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue 0:66 Differential N-Glycosylation Patterns in Lung Adenocarcinoma Tissue. 25658811 1 0 theme protected 152:160 arg1 candidate 264:272 an attractive candidate 250:272 an attractive candidate for the development of immunotherapies 250:311 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 0 theme protected 152:160 arg1 trisaccharide 162:174 an orthogonally protected trisaccharide 136:174 an orthogonally protected trisaccharide 136:174 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 1 theme trisaccharide 162:174 arg1 synthesis 123:131 The chemical synthesis 110:131 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, 110:312 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 2 theme attractive 253:262 arg1 candidate 264:272 an attractive candidate 250:272 an attractive candidate for the development of immunotherapies 250:311 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 2 theme attractive 253:262 arg1 trisaccharide 162:174 an orthogonally protected trisaccharide 136:174 an orthogonally protected trisaccharide 136:174 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 4 3 theme repeating 646:654 arg1 unit 656:659 a spacer-containing repeating unit 626:659 a spacer-containing repeating unit suitable for immunological investigations 626:701 Versatility of the trisaccharide was demonstrated by its transformation into a spacer-containing repeating unit suitable for immunological investigations. 25658811 3 4 theme oxidation 529:537 arg1 approach 539:546 a late stage oxidation approach 516:546 a late stage oxidation approach 516:546 Lactamization of a β-D-ManpNAcA moiety during deprotection was avoided by a late stage oxidation approach. 25658811 4 5 theme spacer-containing 628:644 arg1 unit 656:659 a spacer-containing repeating unit 626:659 a spacer-containing repeating unit suitable for immunological investigations 626:701 Versatility of the trisaccharide was demonstrated by its transformation into a spacer-containing repeating unit suitable for immunological investigations. 25658811 3 6 theme β-D-ManpNAcA 461:472 arg1 moiety 474:479 a β-D-ManpNAcA moiety 459:479 a β-D-ManpNAcA moiety 459:479 Lactamization of a β-D-ManpNAcA moiety during deprotection was avoided by a late stage oxidation approach. 25658811 0 7 theme lactamization 95:107 arg1 problem 84:90 the problem 80:90 the problem of lactamization 80:107 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 1 8 theme aureus 226:231 arg1 Type 233:236 Staphylococcus aureus Type 5 211:238 Staphylococcus aureus Type 5 211:238 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 3 9 theme moiety 474:479 arg1 Lactamization 442:454 Lactamization 442:454 Lactamization of a β-D-ManpNAcA moiety during deprotection 442:499 Lactamization of a β-D-ManpNAcA moiety during deprotection was avoided by a late stage oxidation approach. 25658811 1 10 theme immunotherapies 297:311 arg1 development 282:292 the development 278:292 the development of immunotherapies 278:311 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 2 11 theme careful 405:411 arg1 choice 413:418 the careful choice 401:418 the careful choice of protecting groups 401:439 The challenging α-fucosylation and β-mannosylation are addressed through the careful choice of protecting groups. 25658811 0 12 theme aureus 28:33 arg1 unit 66:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 0 13 theme Staphylococcus 13:26 arg1 unit 66:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 1 14 theme Staphylococcus 211:224 arg1 Type 233:236 Staphylococcus aureus Type 5 211:238 Staphylococcus aureus Type 5 211:238 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 15 theme chemical 114:121 arg1 synthesis 123:131 The chemical synthesis 110:131 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, 110:312 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 16 attach derived 176:182 arg2 trisaccharide 162:174 an orthogonally protected trisaccharide 136:174 an orthogonally protected trisaccharide 136:174 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 16 attach derived 176:182 arg1 polysaccharide 193:206 the polysaccharide 189:206 the polysaccharide of Staphylococcus aureus Type 5 189:238 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 1 16 attach derived 176:182 arg2 candidate 264:272 an attractive candidate 250:272 an attractive candidate for the development of immunotherapies 250:311 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 2 17 theme groups 434:439 arg1 choice 413:418 the careful choice 401:418 the careful choice of protecting groups 401:439 The challenging α-fucosylation and β-mannosylation are addressed through the careful choice of protecting groups. 25658811 0 18 theme Type 35:38 arg1 unit 66:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 1 19 theme Type 233:236 arg1 polysaccharide 193:206 the polysaccharide 189:206 the polysaccharide of Staphylococcus aureus Type 5 189:238 The chemical synthesis of an orthogonally protected trisaccharide derived from the polysaccharide of Staphylococcus aureus Type 5, which is an attractive candidate for the development of immunotherapies, is described. 25658811 0 20 theme repeating 56:64 arg1 unit 66:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 3 21 theme late 518:521 arg1 approach 539:546 a late stage oxidation approach 516:546 a late stage oxidation approach 516:546 Lactamization of a β-D-ManpNAcA moiety during deprotection was avoided by a late stage oxidation approach. 25658811 0 22 theme trisaccharide 42:54 arg1 unit 66:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Staphylococcus aureus Type 5 trisaccharide repeating unit 13:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 25658811 3 23 theme stage 523:527 arg1 approach 539:546 a late stage oxidation approach 516:546 a late stage oxidation approach 516:546 Lactamization of a β-D-ManpNAcA moiety during deprotection was avoided by a late stage oxidation approach. 25658811 4 24 theme trisaccharide 568:580 arg1 Versatility 549:559 Versatility 549:559 Versatility of the trisaccharide 549:580 Versatility of the trisaccharide was demonstrated by its transformation into a spacer-containing repeating unit suitable for immunological investigations. 25658811 2 25 theme challenging 332:342 arg1 α-fucosylation 344:357 α-fucosylation 344:357 α-fucosylation 344:357 The challenging α-fucosylation and β-mannosylation are addressed through the careful choice of protecting groups. 25658811 4 26 theme suitable 661:668 arg1 unit 656:659 a spacer-containing repeating unit 626:659 a spacer-containing repeating unit suitable for immunological investigations 626:701 Versatility of the trisaccharide was demonstrated by its transformation into a spacer-containing repeating unit suitable for immunological investigations. 25658811 2 27 theme protecting 423:432 arg1 groups 434:439 protecting groups 423:439 protecting groups 423:439 The challenging α-fucosylation and β-mannosylation are addressed through the careful choice of protecting groups. 25658811 4 28 theme immunological 674:686 arg1 investigations 688:701 immunological investigations 674:701 immunological investigations 674:701 Versatility of the trisaccharide was demonstrated by its transformation into a spacer-containing repeating unit suitable for immunological investigations. 25658811 0 29 theme unit 66:69 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit 0:69 Synthesis of Staphylococcus aureus Type 5 trisaccharide repeating unit: solving the problem of lactamization. 26966177 6 0 theme microcolony 1041:1051 arg1 formation 1053:1061 microcolony formation 1041:1061 microcolony formation 1041:1061 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 9 1 gly glycosylation 1535:1547 arg1 pilins 1594:1599 paralogous pilins 1583:1599 paralogous pilins 1583:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 1 gly glycosylation 1535:1547 arg1 pilus 1552:1556 pilus assembly 1552:1565 pilus assembly 1552:1565 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 10 2 from growth 1606:1611 arg1 condition 1653:1661 a condition 1651:1661 a condition that decreases AglB glycosylation 1651:1695 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 10 2 from growth 1606:1611 arg1 media 1644:1648 low salt media 1635:1648 low salt media 1635:1648 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 4 3 theme abundant 578:585 arg1 pilins 587:592 the most abundant pilins 569:592 the most abundant pilins 569:592 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 7 4 theme surface 1185:1191 arg1 adhesion 1193:1200 strong surface adhesion 1178:1200 strong surface adhesion 1178:1200 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 9 5 theme paralogous 1583:1592 arg1 pilins 1594:1599 paralogous pilins 1583:1599 paralogous pilins 1583:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 6 6 from PilA6 1005:1009 arg1 ΔaglB 1024:1028 ΔpilA[1-6]ΔaglB 1014:1028 ΔpilA[1-6]ΔaglB 1014:1028 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 4 7 mod modified 638:645 arg3 pentasaccharide 715:729 the same pentasaccharide 706:729 the same pentasaccharide detected on H. volcanii flagellins 706:764 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 7 mod modified 638:645 arg1 pilins 587:592 the most abundant pilins 569:592 the most abundant pilins 569:592 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 10 8 theme cell 1889:1892 arg1 states 1894:1899 sessile cell states 1881:1899 sessile cell states 1881:1899 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 0 9 theme Microcolony 109:119 arg1 Formation 121:129 Microcolony Formation 109:129 Microcolony Formation 109:129 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 2 10 theme flagellins 304:313 arg1 N-glycosylation 285:299 the oligosacharyltransferase (AglB)-dependent N-glycosylation 239:299 the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins 239:313 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 4 11 theme strains 625:631 arg1 pili 597:600 pili 597:600 pili of wild-type and ΔaglB strains 597:631 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 10 12 theme microcolony 1714:1724 arg1 formation 1726:1734 microcolony formation 1714:1734 microcolony formation 1714:1734 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 5 13 theme discrete 835:842 arg1 pili 844:847 discrete pili 835:847 discrete pili 835:847 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 0 14 from Roles 66:70 arg1 Adhesion 95:102 Adhesion 95:102 Adhesion 95:102 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 0 14 from Roles 66:70 arg1 Biosynthesis 81:92 Pilus Biosynthesis 75:92 Pilus Biosynthesis 75:92 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 0 14 from Roles 66:70 arg1 Formation 121:129 Microcolony Formation 109:129 Microcolony Formation 109:129 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 7 15 theme strong 1178:1183 arg1 adhesion 1193:1200 strong surface adhesion 1178:1200 strong surface adhesion 1178:1200 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 0 16 theme Haloferax 18:26 arg1 volcanii 28:35 Haloferax volcanii 18:35 Haloferax volcanii Pilin N-Glycans 18:51 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 0 17 theme Pilus 75:79 arg1 Biosynthesis 81:92 Pilus Biosynthesis 75:92 Pilus Biosynthesis 75:92 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 5 18 theme cells 879:883 arg1 layer 870:874 a dispersed layer 858:874 a dispersed layer of cells 858:883 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 5 19 theme pili 930:933 arg1 bundles 935:941 thick pili bundles 924:941 thick pili bundles 924:941 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 3 20 theme archaeal 456:463 arg1 pili 473:476 the structurally related archaeal type IV pili 431:476 the structurally related archaeal type IV pili 431:476 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 7 21 theme ΔpilA[1-6 1283:1291 arg1 cells 1299:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 5 22 theme ΔaglB 907:911 arg1 cells 913:917 ΔaglB cells 907:917 ΔaglB cells 907:917 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 7 23 theme expressing 1121:1130 arg1 PilA3 1132:1136 expressing PilA3 1121:1136 expressing PilA3 1121:1136 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 3 24 gly N-glycosylation 367:381 arg1 pili 473:476 the structurally related archaeal type IV pili 431:476 the structurally related archaeal type IV pili 431:476 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 3 25 theme IV 470:471 arg1 pili 473:476 the structurally related archaeal type IV pili 431:476 the structurally related archaeal type IV pili 431:476 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 0 26 theme Pilin 37:41 arg1 N-Glycans 43:51 Haloferax volcanii Pilin N-Glycans 18:51 Haloferax volcanii Pilin N-Glycans 18:51 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 4 27 theme Haloferax 516:524 arg1 PilA1 552:556 PilA1 552:556 PilA1 552:556 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 27 theme Haloferax 516:524 arg1 pilins 544:549 six Haloferax volcanii adhesion pilins 512:549 six Haloferax volcanii adhesion pilins 512:549 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 27 theme Haloferax 516:524 arg1 PilA2 562:566 PilA2 562:566 PilA2 562:566 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 8 28 theme ΔpilA[1-6 1348:1356 arg1 cells 1364:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 Although PilA4 assembles into pili in the ΔpilA[1-6]ΔaglB cells, these pili are, unlike wild-type pili, curled, perhaps rendering them non-functional. 26966177 10 29 theme cells 1626:1630 arg1 growth 1606:1611 The growth 1602:1611 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, 1602:1696 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 5 30 theme wild-type 783:791 arg1 cells 793:797 wild-type cells 783:797 wild-type cells 783:797 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 9 31 theme effect 1525:1530 arg1 this 1475:1478 this 1475:1478 this 1475:1478 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 31 theme effect 1525:1530 arg1 demonstration 1493:1505 the first demonstration 1483:1505 the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1483:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 4 32 theme adhesion 535:542 arg1 PilA1 552:556 PilA1 552:556 PilA1 552:556 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 32 theme adhesion 535:542 arg1 pilins 544:549 six Haloferax volcanii adhesion pilins 512:549 six Haloferax volcanii adhesion pilins 512:549 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 32 theme adhesion 535:542 arg1 PilA2 562:566 PilA2 562:566 PilA2 562:566 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 8 33 theme ΔaglB 1358:1362 arg1 cells 1364:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 Although PilA4 assembles into pili in the ΔpilA[1-6]ΔaglB cells, these pili are, unlike wild-type pili, curled, perhaps rendering them non-functional. 26966177 0 34 with Identification 0:13 arg1 Roles 66:70 Diverse Roles 58:70 Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation 58:129 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 9 35 theme glycosylation 1535:1547 arg1 effect 1525:1530 a differential effect 1510:1530 a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1510:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 7 36 theme microcolony 1211:1221 arg1 formation 1223:1231 microcolony formation 1211:1231 microcolony formation 1211:1231 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 9 37 theme pilus 1552:1556 arg1 assembly 1558:1565 pilus assembly 1552:1565 pilus assembly 1552:1565 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 10 38 theme low 1635:1637 arg1 condition 1653:1661 a condition 1651:1661 a condition that decreases AglB glycosylation 1651:1695 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 10 38 theme low 1635:1637 arg1 media 1644:1648 low salt media 1635:1648 low salt media 1635:1648 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 2 39 gly N-glycosylation 285:299 arg1 archaea 230:236 many archaea 225:236 many archaea 225:236 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 2 39 gly N-glycosylation 285:299 arg1 flagella 331:338 flagella assembly 331:347 flagella assembly 331:347 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 2 39 gly N-glycosylation 285:299 arg1 flagellins 304:313 flagellins 304:313 flagellins 304:313 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 2 40 theme -dependent 274:283 arg1 N-glycosylation 285:299 the oligosacharyltransferase (AglB)-dependent N-glycosylation 239:299 the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins 239:313 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 1 41 theme common 185:190 arg1 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 1 41 theme common 185:190 arg1 modification 172:183 a post-translational modification 151:183 a post-translational modification common to all three domains of life 151:219 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 9 42 theme pilins 1594:1599 arg1 function 1571:1578 function 1571:1578 function 1571:1578 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 42 theme pilins 1594:1599 arg1 assembly 1558:1565 pilus assembly 1552:1565 pilus assembly 1552:1565 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 4 43 from pilins 587:592 arg1 pili 597:600 pili 597:600 pili of wild-type and ΔaglB strains 597:631 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 6 44 from PilA2 995:999 arg1 ΔaglB 1024:1028 ΔpilA[1-6]ΔaglB 1014:1028 ΔpilA[1-6]ΔaglB 1014:1028 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 8 45 from pili 1336:1339 arg1 cells 1364:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 the ΔpilA[1-6]ΔaglB cells 1344:1368 Although PilA4 assembles into pili in the ΔpilA[1-6]ΔaglB cells, these pili are, unlike wild-type pili, curled, perhaps rendering them non-functional. 26966177 5 46 contain have 919:922 arg1 cells 913:917 ΔaglB cells 907:917 ΔaglB cells 907:917 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 5 46 contain have 919:922 arg2 bundles 935:941 thick pili bundles 924:941 thick pili bundles 924:941 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 10 47 theme AglB 1678:1681 arg1 glycosylation 1683:1695 AglB glycosylation 1678:1695 AglB glycosylation 1678:1695 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 7 48 from PilA4 1141:1145 arg1 cells 1161:1165 ΔpilA[1-6] cells 1150:1165 ΔpilA[1-6] cells 1150:1165 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 4 49 theme wild-type 605:613 arg1 strains 625:631 wild-type and ΔaglB strains 605:631 wild-type and ΔaglB strains 605:631 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 10 50 theme sessile 1881:1887 arg1 states 1894:1899 sessile cell states 1881:1899 sessile cell states 1881:1899 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 1 51 theme life 216:219 arg1 life 216:219 life 216:219 life 216:219 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 1 51 theme life 216:219 arg1 domains 205:211 all three domains 195:211 all three domains of life 195:219 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 9 52 from function 1571:1578 arg1 this 1475:1478 this 1475:1478 this 1475:1478 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 52 from function 1571:1578 arg1 demonstration 1493:1505 the first demonstration 1483:1505 the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1483:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 4 53 theme ΔaglB 619:623 arg1 strains 625:631 wild-type and ΔaglB strains 605:631 wild-type and ΔaglB strains 605:631 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 2 54 theme many 225:228 arg1 archaea 230:236 many archaea 225:236 many archaea 225:236 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 9 55 from assembly 1558:1565 arg1 this 1475:1478 this 1475:1478 this 1475:1478 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 55 from assembly 1558:1565 arg1 demonstration 1493:1505 the first demonstration 1483:1505 the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1483:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 8 56 theme wild-type 1394:1402 arg1 pili 1404:1407 wild-type pili 1394:1407 wild-type pili 1394:1407 Although PilA4 assembles into pili in the ΔpilA[1-6]ΔaglB cells, these pili are, unlike wild-type pili, curled, perhaps rendering them non-functional. 26966177 4 57 theme AglB-dependent 681:694 arg1 manner 696:701 an AglB-dependent manner 678:701 an AglB-dependent manner 678:701 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 5 58 theme plastic 890:896 arg1 surface 898:904 a plastic surface 888:904 a plastic surface 888:904 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 9 59 from demonstration 1493:1505 arg1 function 1571:1578 function 1571:1578 function 1571:1578 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 59 from demonstration 1493:1505 arg1 assembly 1558:1565 pilus assembly 1552:1565 pilus assembly 1552:1565 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 6 60 from expression 1083:1092 arg1 ΔpilA[1-6 1097:1105 ΔpilA[1-6 1097:1105 ΔpilA[1-6 1097:1105 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 5 61 theme dispersed 860:868 arg1 layer 870:874 a dispersed layer 858:874 a dispersed layer of cells 858:883 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 6 62 theme expressing 977:986 arg1 PilA1 988:992 expressing PilA1 977:992 expressing PilA1 977:992 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 7 63 theme ΔaglB 1293:1297 arg1 cells 1299:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 4 64 theme planktonic 653:662 arg1 conditions 664:673 planktonic conditions 653:673 planktonic conditions 653:673 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 1 65 theme post-translational 153:170 arg1 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation 132:146 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 1 65 theme post-translational 153:170 arg1 modification 172:183 a post-translational modification 151:183 a post-translational modification common to all three domains of life 151:219 N-Glycosylation is a post-translational modification common to all three domains of life. 26966177 6 66 from PilA1 988:992 arg1 ΔaglB 1024:1028 ΔpilA[1-6]ΔaglB 1014:1028 ΔpilA[1-6]ΔaglB 1014:1028 Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. 26966177 5 67 contain have 806:809 arg2 surfaces 811:818 surfaces 811:818 surfaces decorated with discrete pili 811:847 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 5 67 contain have 806:809 arg1 cells 793:797 wild-type cells 783:797 wild-type cells 783:797 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 7 68 theme ΔpilA[1-6 1150:1158 arg1 cells 1161:1165 ΔpilA[1-6] cells 1150:1165 ΔpilA[1-6] cells 1150:1165 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 5 69 theme thick 924:928 arg1 bundles 935:941 thick pili bundles 924:941 thick pili bundles 924:941 However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. 26966177 3 70 dep assembly 403:410 arg1 the 399:401 the 399:401 the 399:401 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 4 71 theme H. 743:744 arg1 flagellins 755:764 H. volcanii flagellins 743:764 H. volcanii flagellins 743:764 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 7 72 from PilA3 1132:1136 arg1 cells 1161:1165 ΔpilA[1-6] cells 1150:1165 ΔpilA[1-6] cells 1150:1165 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 3 73 theme type 465:468 arg1 pili 473:476 the structurally related archaeal type IV pili 431:476 the structurally related archaeal type IV pili 431:476 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 7 74 theme surface 1263:1269 arg1 adhesion 1271:1278 surface adhesion 1263:1278 surface adhesion in ΔpilA[1-6]ΔaglB cells 1263:1303 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 4 75 theme same 710:713 arg1 pentasaccharide 715:729 the same pentasaccharide 706:729 the same pentasaccharide detected on H. volcanii flagellins 706:764 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 3 76 theme related 448:454 arg1 pili 473:476 the structurally related archaeal type IV pili 431:476 the structurally related archaeal type IV pili 431:476 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 0 77 theme volcanii 28:35 arg1 N-Glycans 43:51 Haloferax volcanii Pilin N-Glycans 18:51 Haloferax volcanii Pilin N-Glycans 18:51 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 9 78 theme first 1487:1491 arg1 this 1475:1478 this 1475:1478 this 1475:1478 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 78 theme first 1487:1491 arg1 demonstration 1493:1505 the first demonstration 1483:1505 the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1483:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 79 from effect 1525:1530 arg1 function 1571:1578 function 1571:1578 function 1571:1578 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 9 79 from effect 1525:1530 arg1 assembly 1558:1565 pilus assembly 1552:1565 pilus assembly 1552:1565 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 0 80 theme N-Glycans 43:51 arg1 Identification 0:13 Identification 0:13 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 0:130 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 4 81 theme volcanii 526:533 arg1 PilA1 552:556 PilA1 552:556 PilA1 552:556 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 81 theme volcanii 526:533 arg1 pilins 544:549 six Haloferax volcanii adhesion pilins 512:549 six Haloferax volcanii adhesion pilins 512:549 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 81 theme volcanii 526:533 arg1 PilA2 562:566 PilA2 562:566 PilA2 562:566 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 3 82 theme pili 473:476 arg1 assembly 403:410 assembly 403:410 assembly 403:410 However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. 26966177 0 83 theme Diverse 58:64 arg1 Roles 66:70 Diverse Roles 58:70 Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation 58:129 Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation. 26966177 9 84 theme differential 1512:1523 arg1 effect 1525:1530 a differential effect 1510:1530 a differential effect of glycosylation on pilus assembly and function of paralogous pilins 1510:1599 To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. 26966177 4 85 located detected 731:738 arg1 flagellins 755:764 H. volcanii flagellins 743:764 H. volcanii flagellins 743:764 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 4 85 located detected 731:738 arg2 pentasaccharide 715:729 the same pentasaccharide 706:729 the same pentasaccharide detected on H. volcanii flagellins 706:764 Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. 26966177 10 86 theme important 1815:1823 arg1 role 1825:1828 an important role 1812:1828 an important role 1812:1828 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 10 87 theme wild-type 1616:1624 arg1 cells 1626:1630 wild-type cells 1616:1630 wild-type cells 1616:1630 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 2 88 theme flagella 331:338 arg1 assembly 340:347 flagella assembly 331:347 flagella assembly 331:347 In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. 26966177 7 89 from adhesion 1271:1278 arg1 cells 1299:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 ΔpilA[1-6]ΔaglB cells 1283:1303 Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. 26966177 10 90 theme salt 1639:1642 arg1 condition 1653:1661 a condition 1651:1661 a condition that decreases AglB glycosylation 1651:1695 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 26966177 10 90 theme salt 1639:1642 arg1 media 1644:1648 low salt media 1635:1648 low salt media 1635:1648 The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress. 24275651 0 0 theme oligosaccharyltransferase 68:92 arg1 PglB 94:97 the bacterial oligosaccharyltransferase PglB 54:97 the bacterial oligosaccharyltransferase PglB 54:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 7 1 from roles 1160:1164 arg1 glycosylation 1190:1202 OST-catalyzed glycosylation 1176:1202 OST-catalyzed glycosylation 1176:1202 Our findings define two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation. 24275651 6 2 theme essential 1007:1015 arg1 disengagement 947:959 disengagement 947:959 disengagement of the structurally ordered part of EL5 947:999 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 6 2 theme essential 1007:1015 arg1 step 1017:1020 an essential step 1004:1020 an essential step of the glycosylation reaction 1004:1050 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 0 3 theme bacterial 58:66 arg1 PglB 94:97 the bacterial oligosaccharyltransferase PglB 54:97 the bacterial oligosaccharyltransferase PglB 54:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 7 4 theme distinct 1139:1146 arg1 roles 1160:1164 two distinct mechanistic roles 1135:1164 two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation 1135:1202 Our findings define two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation. 24275651 6 5 theme glyco-product 1092:1104 arg1 release 1106:1112 glyco-product release 1092:1112 glyco-product release 1092:1112 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 3 6 from role 547:550 arg1 catalysis 555:563 catalysis 555:563 catalysis 555:563 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 8 7 theme slight 1339:1344 arg1 reduction 1346:1354 only slight reduction 1334:1354 only slight reduction in activity 1334:1366 These functions, exerted by the two halves of EL5, are independent, because the loop can be cleaved by specific proteolysis with only slight reduction in activity. 24275651 4 8 theme conserved 660:668 arg1 Tyr293 688:693 Tyr293 688:693 Tyr293 688:693 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 8 theme conserved 660:668 arg1 residue 679:685 a conserved tyrosine residue 658:685 a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis 658:747 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 9 theme new 594:596 arg1 motif 634:638 a new and functionally important sequence motif 592:638 a new and functionally important sequence motif 592:638 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 0 10 theme PglB 94:97 arg1 loop 44:47 external loop 5 35:49 external loop 5 of the bacterial oligosaccharyltransferase PglB 35:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 6 11 theme disulfide 909:917 arg1 cross-linking 919:931 site-directed disulfide cross-linking 895:931 site-directed disulfide cross-linking 895:931 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 2 12 theme membrane 335:342 arg1 oligosaccharyltransferase 352:376 the integral membrane protein oligosaccharyltransferase 322:376 the integral membrane protein oligosaccharyltransferase (OST) 322:382 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 2 12 theme membrane 335:342 arg1 OST 379:381 OST 379:381 OST 379:381 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 4 13 theme side 711:714 arg1 chain 716:720 chain 716:720 chain 716:720 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 2 14 theme acceptor 285:292 arg1 asparagines 294:304 acceptor asparagines 285:304 acceptor asparagines 285:304 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 3 15 theme Campylobacter 455:467 arg1 PglB 482:485 the Campylobacter lari protein PglB 451:485 the Campylobacter lari protein PglB 451:485 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 15 theme Campylobacter 455:467 arg1 enzyme 443:448 a single-subunit OST enzyme 422:448 a single-subunit OST enzyme 422:448 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 6 16 theme EL5 997:999 arg1 part 989:992 the structurally ordered part 964:992 the structurally ordered part of EL5 964:999 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 0 17 theme external 35:42 arg1 loop 44:47 external loop 5 35:49 external loop 5 of the bacterial oligosaccharyltransferase PglB 35:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 3 18 theme lari 469:472 arg1 PglB 482:485 the Campylobacter lari protein PglB 451:485 the Campylobacter lari protein PglB 451:485 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 18 theme lari 469:472 arg1 enzyme 443:448 a single-subunit OST enzyme 422:448 a single-subunit OST enzyme 422:448 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 2 19 theme integral 326:333 arg1 oligosaccharyltransferase 352:376 the integral membrane protein oligosaccharyltransferase 322:376 the integral membrane protein oligosaccharyltransferase (OST) 322:382 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 2 19 theme integral 326:333 arg1 OST 379:381 OST 379:381 OST 379:381 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 6 20 theme site-directed 895:907 arg1 cross-linking 919:931 site-directed disulfide cross-linking 895:931 site-directed disulfide cross-linking 895:931 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 6 21 theme sequon 1074:1079 arg1 binding 1081:1087 sequon binding 1074:1087 sequon binding 1074:1087 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 7 22 theme EL5 1169:1171 arg1 roles 1160:1164 two distinct mechanistic roles 1135:1164 two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation 1135:1202 Our findings define two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation. 24275651 8 23 theme EL5 1251:1253 arg1 halves 1241:1246 the two halves 1233:1246 the two halves of EL5 1233:1253 These functions, exerted by the two halves of EL5, are independent, because the loop can be cleaved by specific proteolysis with only slight reduction in activity. 24275651 5 24 theme mutated 860:866 arg1 PglB 868:871 mutated PglB 860:871 mutated PglB lacking Tyr293 860:886 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 4 25 theme tyrosine 670:677 arg1 Tyr293 688:693 Tyr293 688:693 Tyr293 688:693 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 25 theme tyrosine 670:677 arg1 residue 679:685 a conserved tyrosine residue 658:685 a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis 658:747 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 6 26 theme part 989:992 arg1 disengagement 947:959 disengagement 947:959 disengagement of the structurally ordered part of EL5 947:999 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 6 26 theme part 989:992 arg1 step 1017:1020 an essential step 1004:1020 an essential step of the glycosylation reaction 1004:1050 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 1 27 theme Asparagine-linked 100:116 arg1 modification 164:175 a post-translational protein modification 135:175 a post-translational protein modification that is conserved in all domains of life 135:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 27 theme Asparagine-linked 100:116 arg1 glycosylation 118:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 28 theme life 213:216 arg1 domains 202:208 all domains 198:208 all domains of life 198:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 28 theme life 213:216 arg1 life 213:216 life 213:216 life 213:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 0 29 theme essential 16:24 arg1 motif 26:30 A catalytically essential motif 0:30 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB 0:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 6 30 theme ordered 981:987 arg1 part 989:992 the structurally ordered part 964:992 the structurally ordered part of EL5 964:999 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 3 31 theme reported 400:407 arg1 structure 409:417 The previously reported structure 385:417 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, 385:486 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 32 theme OST 439:441 arg1 PglB 482:485 the Campylobacter lari protein PglB 451:485 the Campylobacter lari protein PglB 451:485 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 32 theme OST 439:441 arg1 enzyme 443:448 a single-subunit OST enzyme 422:448 a single-subunit OST enzyme 422:448 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 4 33 theme aromatic 702:709 arg1 chain 716:720 chain 716:720 chain 716:720 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 34 contain containing 647:656 arg1 EL5 643:645 EL5 643:645 EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis 643:747 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 34 contain containing 647:656 arg2 residue 679:685 a conserved tyrosine residue 658:685 a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis 658:747 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 4 34 contain containing 647:656 arg2 Tyr293 688:693 Tyr293 688:693 Tyr293 688:693 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 2 35 theme lipid-linked 245:256 arg1 LLO 275:277 LLO 275:277 LLO 275:277 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 2 35 theme lipid-linked 245:256 arg1 oligosaccharide 258:272 a lipid-linked oligosaccharide 243:272 a lipid-linked oligosaccharide (LLO) 243:278 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 3 36 theme protein 474:480 arg1 PglB 482:485 the Campylobacter lari protein PglB 451:485 the Campylobacter lari protein PglB 451:485 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 36 theme protein 474:480 arg1 enzyme 443:448 a single-subunit OST enzyme 422:448 a single-subunit OST enzyme 422:448 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 37 theme enzyme 443:448 arg1 structure 409:417 The previously reported structure 385:417 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, 385:486 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 5 38 theme conserved 785:793 arg1 motif 795:799 the conserved motif 781:799 the conserved motif 781:799 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 4 39 theme sequence 625:632 arg1 motif 634:638 a new and functionally important sequence motif 592:638 a new and functionally important sequence motif 592:638 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 24275651 6 40 theme reaction 1043:1050 arg1 disengagement 947:959 disengagement 947:959 disengagement of the structurally ordered part of EL5 947:999 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 6 40 theme reaction 1043:1050 arg1 step 1017:1020 an essential step 1004:1020 an essential step of the glycosylation reaction 1004:1050 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 7 41 theme mechanistic 1148:1158 arg1 roles 1160:1164 two distinct mechanistic roles 1135:1164 two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation 1135:1202 Our findings define two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation. 24275651 3 42 theme disordered 509:518 arg1 EL5 535:537 EL5 535:537 EL5 535:537 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 42 theme disordered 509:518 arg1 loop 529:532 a partially disordered external loop 497:532 a partially disordered external loop (EL5) 497:538 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 43 theme single-subunit 424:437 arg1 PglB 482:485 the Campylobacter lari protein PglB 451:485 the Campylobacter lari protein PglB 451:485 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 43 theme single-subunit 424:437 arg1 enzyme 443:448 a single-subunit OST enzyme 422:448 a single-subunit OST enzyme 422:448 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 5 44 contain containing 770:779 arg1 peptide 762:768 A synthetic peptide 750:768 A synthetic peptide containing the conserved motif 750:799 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 5 44 contain containing 770:779 arg2 motif 795:799 the conserved motif 781:799 the conserved motif 781:799 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 2 45 theme initial 223:229 arg1 transfer 231:238 The initial transfer 219:238 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines 219:304 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 3 46 theme external 520:527 arg1 EL5 535:537 EL5 535:537 EL5 535:537 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 3 46 theme external 520:527 arg1 loop 529:532 a partially disordered external loop 497:532 a partially disordered external loop (EL5) 497:538 The previously reported structure of a single-subunit OST enzyme, the Campylobacter lari protein PglB, revealed a partially disordered external loop (EL5), whose role in catalysis was unclear. 24275651 8 47 with proteolysis 1317:1327 arg1 reduction 1346:1354 only slight reduction 1334:1354 only slight reduction in activity 1334:1366 These functions, exerted by the two halves of EL5, are independent, because the loop can be cleaved by specific proteolysis with only slight reduction in activity. 24275651 1 48 theme post-translational 137:154 arg1 modification 164:175 a post-translational protein modification 135:175 a post-translational protein modification that is conserved in all domains of life 135:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 48 theme post-translational 137:154 arg1 glycosylation 118:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 8 49 from reduction 1346:1354 arg1 activity 1359:1366 activity 1359:1366 activity 1359:1366 These functions, exerted by the two halves of EL5, are independent, because the loop can be cleaved by specific proteolysis with only slight reduction in activity. 24275651 6 50 theme glycosylation 1029:1041 arg1 reaction 1043:1050 the glycosylation reaction 1025:1050 the glycosylation reaction 1025:1050 Using site-directed disulfide cross-linking, we show that disengagement of the structurally ordered part of EL5 is an essential step of the glycosylation reaction, probably by allowing sequon binding or glyco-product release. 24275651 1 51 theme protein 156:162 arg1 modification 164:175 a post-translational protein modification 135:175 a post-translational protein modification that is conserved in all domains of life 135:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 51 theme protein 156:162 arg1 glycosylation 118:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 2 52 theme protein 344:350 arg1 oligosaccharyltransferase 352:376 the integral membrane protein oligosaccharyltransferase 322:376 the integral membrane protein oligosaccharyltransferase (OST) 322:382 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 2 52 theme protein 344:350 arg1 OST 379:381 OST 379:381 OST 379:381 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 1 53 link Asparagine-linked 100:116 arg1 modification 164:175 a post-translational protein modification 135:175 a post-translational protein modification that is conserved in all domains of life 135:216 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 1 53 link Asparagine-linked 100:116 arg1 glycosylation 118:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation 100:130 Asparagine-linked glycosylation is a post-translational protein modification that is conserved in all domains of life. 24275651 5 54 dep in 839:840 arg1 vitro 842:846 vitro 842:846 vitro 842:846 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 5 55 theme in 839:840 arg1 activity 848:855 in vitro activity 839:855 in vitro activity of mutated PglB lacking Tyr293 839:886 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 8 56 theme specific 1308:1315 arg1 proteolysis 1317:1327 specific proteolysis 1308:1327 specific proteolysis with only slight reduction in activity 1308:1366 These functions, exerted by the two halves of EL5, are independent, because the loop can be cleaved by specific proteolysis with only slight reduction in activity. 24275651 5 57 theme PglB 868:871 arg1 activity 848:855 in vitro activity 839:855 in vitro activity of mutated PglB lacking Tyr293 839:886 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 2 58 link lipid-linked 245:256 arg1 LLO 275:277 LLO 275:277 LLO 275:277 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 2 58 link lipid-linked 245:256 arg1 oligosaccharide 258:272 a lipid-linked oligosaccharide 243:272 a lipid-linked oligosaccharide (LLO) 243:278 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 5 59 theme synthetic 752:760 arg1 peptide 762:768 A synthetic peptide 750:768 A synthetic peptide containing the conserved motif 750:799 A synthetic peptide containing the conserved motif can partially but specifically rescue in vitro activity of mutated PglB lacking Tyr293. 24275651 0 60 from motif 26:30 arg1 loop 44:47 external loop 5 35:49 external loop 5 of the bacterial oligosaccharyltransferase PglB 35:97 A catalytically essential motif in external loop 5 of the bacterial oligosaccharyltransferase PglB. 24275651 2 61 theme oligosaccharide 258:272 arg1 transfer 231:238 The initial transfer 219:238 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines 219:304 The initial transfer of a lipid-linked oligosaccharide (LLO) onto acceptor asparagines is catalyzed by the integral membrane protein oligosaccharyltransferase (OST). 24275651 7 62 theme OST-catalyzed 1176:1188 arg1 glycosylation 1190:1202 OST-catalyzed glycosylation 1176:1202 OST-catalyzed glycosylation 1176:1202 Our findings define two distinct mechanistic roles of EL5 in OST-catalyzed glycosylation. 24275651 4 63 theme important 615:623 arg1 motif 634:638 a new and functionally important sequence motif 592:638 a new and functionally important sequence motif 592:638 We identified a new and functionally important sequence motif in EL5 containing a conserved tyrosine residue (Tyr293) whose aromatic side chain is essential for catalysis. 25746926 2 0 theme kills 512:516 arg1 parasites 518:526 kills parasites 512:526 kills parasites 512:526 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 3 1 theme paucimannose 740:751 arg1 enrichment 714:723 an enrichment 711:723 an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures 711:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 4 2 from modifications 893:905 arg1 composition 919:929 N-glycan composition 910:929 N-glycan composition 910:929 Two molecular mechanisms involved in resistance were identified: VSG switching and modifications in N-glycan composition. 25746926 1 3 theme surface 181:187 arg1 VSG 204:206 VSG 204:206 VSG 204:206 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 3 theme surface 181:187 arg1 glycoproteins 189:201 Trypanosoma brucei variant surface glycoproteins 154:201 Trypanosoma brucei variant surface glycoproteins (VSG) 154:207 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 2 4 theme impaired 578:585 arg1 cytokinesis 587:597 impaired cytokinesis 578:597 impaired cytokinesis 578:597 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 4 5 theme VSG 875:877 arg1 switching 879:887 VSG switching 875:887 VSG switching 875:887 Two molecular mechanisms involved in resistance were identified: VSG switching and modifications in N-glycan composition. 25746926 2 6 with lectin 403:408 arg1 specificity 462:472 predominant N-acetylglucosamine-binding specificity 422:472 predominant N-acetylglucosamine-binding specificity 422:472 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 5 7 theme responsible 1093:1103 arg1 genes 1025:1029 the TbSTT3A and/or TbSTT3B genes 998:1029 the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity 998:1126 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 4 8 theme molecular 814:822 arg1 mechanisms 824:833 Two molecular mechanisms 810:833 Two molecular mechanisms involved in resistance 810:856 Two molecular mechanisms involved in resistance were identified: VSG switching and modifications in N-glycan composition. 25746926 6 9 theme substantial 1259:1269 arg1 cost 1279:1282 a substantial fitness cost 1257:1282 a substantial fitness cost 1257:1282 Furthermore, UDA-resistant trypanosomes exhibited severely impaired infectivity indicating that the resistant phenotype entails a substantial fitness cost. 25746926 0 10 theme glycosylation 98:110 arg1 defects 112:118 VSG switching and glycosylation defects 80:118 defects 112:118 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 3 11 theme reduction 759:767 arg1 enrichment 714:723 an enrichment 711:723 an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures 711:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 6 12 theme resistant 1229:1237 arg1 phenotype 1239:1247 the resistant phenotype 1225:1247 the resistant phenotype 1225:1247 Furthermore, UDA-resistant trypanosomes exhibited severely impaired infectivity indicating that the resistant phenotype entails a substantial fitness cost. 25746926 6 13 theme impaired 1188:1195 arg1 infectivity 1197:1207 severely impaired infectivity 1179:1207 severely impaired infectivity 1179:1207 Furthermore, UDA-resistant trypanosomes exhibited severely impaired infectivity indicating that the resistant phenotype entails a substantial fitness cost. 25746926 1 14 gly glycosylated 213:224 arg1 VSG 204:206 VSG 204:206 VSG 204:206 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 14 gly glycosylated 213:224 arg1 glycoproteins 189:201 Trypanosoma brucei variant surface glycoproteins 154:201 Trypanosoma brucei variant surface glycoproteins (VSG) 154:207 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 7 15 theme carbohydrate-binding 1522:1541 arg1 agents 1543:1548 carbohydrate-binding agents 1522:1548 carbohydrate-binding agents 1522:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 5 16 theme Glycosylation 932:944 arg1 defects 946:952 Glycosylation defects 932:952 Glycosylation defects 932:952 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 3 17 theme oligomannose 785:796 arg1 structures 798:807 triantennary oligomannose structures 772:807 triantennary oligomannose structures 772:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 7 18 theme glycan 1350:1355 arg1 composition 1357:1367 surface glycan composition 1342:1367 surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1342:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 7 19 theme composition 1357:1367 arg1 modification 1326:1337 the modification 1322:1337 the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1322:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 5 20 theme TbSTT3B 1017:1023 arg1 genes 1025:1029 the TbSTT3A and/or TbSTT3B genes 998:1029 the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity 998:1126 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 2 21 theme nettle 396:401 arg1 lectin 403:408 the stinging nettle lectin 383:408 the stinging nettle lectin (UDA) 383:414 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 2 21 theme nettle 396:401 arg1 UDA 411:413 UDA 411:413 UDA 411:413 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 3 22 theme structures 798:807 arg1 paucimannose 740:751 biantennary paucimannose 728:751 biantennary paucimannose 728:751 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 3 22 theme structures 798:807 arg1 reduction 759:767 a reduction 757:767 a reduction of triantennary oligomannose structures 757:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 4 23 theme N-glycan 910:917 arg1 composition 919:929 N-glycan composition 910:929 N-glycan composition 910:929 Two molecular mechanisms involved in resistance were identified: VSG switching and modifications in N-glycan composition. 25746926 5 24 theme genes 1025:1029 arg1 down-regulation 979:993 the down-regulation 975:993 the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity 975:1126 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 2 25 theme stinging 387:394 arg1 lectin 403:408 the stinging nettle lectin 383:408 the stinging nettle lectin (UDA) 383:414 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 2 25 theme stinging 387:394 arg1 UDA 411:413 UDA 411:413 UDA 411:413 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 0 26 theme brucei 24:29 arg1 Exposure 0:7 Exposure 0:7 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin 0:70 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 6 27 theme fitness 1271:1277 arg1 cost 1279:1282 a substantial fitness cost 1257:1282 a substantial fitness cost 1257:1282 Furthermore, UDA-resistant trypanosomes exhibited severely impaired infectivity indicating that the resistant phenotype entails a substantial fitness cost. 25746926 3 28 theme parasite 634:641 arg1 resistance 643:652 parasite resistance 634:652 parasite resistance 634:652 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 6 29 theme UDA-resistant 1142:1154 arg1 trypanosomes 1156:1167 UDA-resistant trypanosomes 1142:1167 UDA-resistant trypanosomes 1142:1167 Furthermore, UDA-resistant trypanosomes exhibited severely impaired infectivity indicating that the resistant phenotype entails a substantial fitness cost. 25746926 2 30 theme predominant 422:432 arg1 specificity 462:472 predominant N-acetylglucosamine-binding specificity 422:472 predominant N-acetylglucosamine-binding specificity 422:472 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 3 31 theme triantennary 772:783 arg1 structures 798:807 triantennary oligomannose structures 772:807 triantennary oligomannose structures 772:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 0 32 theme reduced 133:139 arg1 infectivity 141:151 reduced infectivity 133:151 reduced infectivity 133:151 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 3 33 theme Prolonged 600:608 arg1 exposure 610:617 Prolonged exposure 600:617 Prolonged exposure to UDA 600:624 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 7 34 theme resistance 1464:1473 arg1 mechanism 1475:1483 a general resistance mechanism 1454:1483 a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1454:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 1 35 theme protective 316:325 arg1 barrier 327:333 a protective barrier 314:333 a protective barrier against the immune system 314:359 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 2 36 gly glycosylated 490:501 arg1 VSGs 503:506 glycosylated VSGs 490:506 glycosylated VSGs 490:506 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 2 37 theme glycosylated 490:501 arg1 VSGs 503:506 glycosylated VSGs 490:506 glycosylated VSGs 490:506 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 1 38 theme barrier 327:333 arg1 formation 301:309 the formation 297:309 the formation of a protective barrier against the immune system 297:359 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 2 39 theme N-acetylglucosamine-binding 434:460 arg1 specificity 462:472 predominant N-acetylglucosamine-binding specificity 422:472 predominant N-acetylglucosamine-binding specificity 422:472 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 5 40 theme glycan 1109:1114 arg1 specificity 1116:1126 glycan specificity 1109:1126 glycan specificity 1109:1126 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 4 41 from switching 879:887 arg1 composition 919:929 N-glycan composition 910:929 N-glycan composition 910:929 Two molecular mechanisms involved in resistance were identified: VSG switching and modifications in N-glycan composition. 25746926 0 42 theme N-acetylglucosamine-binding 37:63 arg1 lectin 65:70 an N-acetylglucosamine-binding lectin 34:70 an N-acetylglucosamine-binding lectin 34:70 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 2 43 from defects 541:547 arg1 endocytosis 552:562 endocytosis 552:562 endocytosis 552:562 Here, we report that the stinging nettle lectin (UDA), with predominant N-acetylglucosamine-binding specificity, interacts with glycosylated VSGs and kills parasites by provoking defects in endocytosis together with impaired cytokinesis. 25746926 1 44 theme immune 347:352 arg1 system 354:359 the immune system 343:359 the immune system 343:359 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 45 gly glycoproteins 189:201 arg1 VSG 204:206 VSG 204:206 VSG 204:206 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 45 gly glycoproteins 189:201 arg1 glycoproteins 189:201 Trypanosoma brucei variant surface glycoproteins 154:201 Trypanosoma brucei variant surface glycoproteins (VSG) 154:207 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 5 46 theme TbSTT3A 1002:1008 arg1 genes 1025:1029 the TbSTT3A and/or TbSTT3B genes 998:1029 the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity 998:1126 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 7 47 theme surface 1342:1348 arg1 composition 1357:1367 surface glycan composition 1342:1367 surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1342:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 3 48 theme diminished 665:674 arg1 capacity 676:683 a diminished capacity 663:683 a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures 663:807 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 7 49 from mechanism 1475:1483 arg1 response 1488:1495 response 1488:1495 response to prolonged exposure to carbohydrate-binding agents 1488:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 5 50 dep oligosaccharyltransferases 1043:1068 arg1 oligosaccharyltransferases 1043:1068 oligosaccharyltransferases A and B 1043:1076 oligosaccharyltransferases A and B 1043:1076 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 5 50 dep oligosaccharyltransferases 1043:1068 arg1 B 1076:1076 B 1076:1076 B 1076:1076 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 5 50 dep oligosaccharyltransferases 1043:1068 arg1 A 1070:1070 A 1070:1070 A 1070:1070 Glycosylation defects were correlated with the down-regulation of the TbSTT3A and/or TbSTT3B genes (coding for oligosaccharyltransferases A and B, respectively) responsible for glycan specificity. 25746926 1 51 theme oligomannose 251:262 arg1 structures 264:273 oligomannose structures 251:273 oligomannose structures 251:273 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 0 52 theme VSG 80:82 arg1 switching 84:92 VSG switching and glycosylation defects 80:118 switching 84:92 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 7 53 theme prolonged 1500:1508 arg1 exposure 1510:1517 prolonged exposure 1500:1517 prolonged exposure to carbohydrate-binding agents 1500:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 7 54 theme general 1456:1462 arg1 mechanism 1475:1483 a general resistance mechanism 1454:1483 a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1454:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 1 55 theme brucei 166:171 arg1 VSG 204:206 VSG 204:206 VSG 204:206 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 55 theme brucei 166:171 arg1 glycoproteins 189:201 Trypanosoma brucei variant surface glycoproteins 154:201 Trypanosoma brucei variant surface glycoproteins (VSG) 154:207 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 3 56 theme biantennary 728:738 arg1 paucimannose 740:751 biantennary paucimannose 728:751 biantennary paucimannose 728:751 Prolonged exposure to UDA induced parasite resistance based on a diminished capacity to bind the lectin due to an enrichment of biantennary paucimannose and a reduction of triantennary oligomannose structures. 25746926 0 57 theme Trypanosoma 12:22 arg1 brucei 24:29 Trypanosoma brucei 12:29 Trypanosoma brucei 12:29 Exposure of Trypanosoma brucei to an N-acetylglucosamine-binding lectin induces VSG switching and glycosylation defects resulting in reduced infectivity. 25746926 7 58 theme genes 1407:1411 arg1 down-regulation 1384:1398 down-regulation 1384:1398 down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents 1384:1548 The results obtained further support the modification of surface glycan composition resulting from down-regulation of the genes coding for oligosaccharyltransferases as a general resistance mechanism in response to prolonged exposure to carbohydrate-binding agents. 25746926 1 59 theme variant 173:179 arg1 VSG 204:206 VSG 204:206 VSG 204:206 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25746926 1 59 theme variant 173:179 arg1 glycoproteins 189:201 Trypanosoma brucei variant surface glycoproteins 154:201 Trypanosoma brucei variant surface glycoproteins (VSG) 154:207 Trypanosoma brucei variant surface glycoproteins (VSG) are glycosylated by both paucimannose and oligomannose structures which are involved in the formation of a protective barrier against the immune system. 25325938 1 0 located found 136:140 arg2 Glycosaminoglycans 61:78 Glycosaminoglycans 61:78 Glycosaminoglycans (GAGs) 61:85 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 1 0 located found 136:140 arg2 carbohydrates 110:122 important sulfated carbohydrates 91:122 important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions 91:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 1 0 located found 136:140 arg1 matrix 163:168 the extracellular matrix 145:168 the extracellular matrix that serve many biological functions 145:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 1 1 theme many 181:184 arg1 functions 197:205 many biological functions 181:205 many biological functions 181:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 4 2 theme photolabile 504:514 arg1 linker 516:521 a photolabile linker 502:521 a photolabile linker 502:521 The oligosaccharides are prepared on a solid support that is equipped with a photolabile linker. 25325938 3 3 theme automated 363:371 arg1 synthesis 373:381 the automated synthesis 359:381 the automated synthesis of two chondroitin sulfate hexasaccharides 359:424 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 3 3 theme automated 363:371 arg1 Described 346:354 Described 346:354 Described 346:354 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 5 4 theme linker 528:533 arg1 cleavage 535:542 The linker cleavage 524:542 The linker cleavage from the resin 524:557 The linker cleavage from the resin is performed in a continuous-flow photoreactor under chemically mild conditions. 25325938 1 5 theme biological 186:195 arg1 functions 197:205 many biological functions 181:205 many biological functions 181:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 6 6 theme described 644:652 arg1 scheme 687:692 a general scheme 677:692 a general scheme 677:692 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 25325938 6 6 theme described 644:652 arg1 approach 654:661 The described approach 640:661 The described approach 640:661 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 25325938 6 7 theme families 747:754 arg1 oligosaccharides 719:734 oligosaccharides 719:734 oligosaccharides of all GAG families 719:754 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 25325938 3 8 theme hexasaccharides 410:424 arg1 synthesis 373:381 the automated synthesis 359:381 the automated synthesis of two chondroitin sulfate hexasaccharides 359:424 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 3 8 theme hexasaccharides 410:424 arg1 Described 346:354 Described 346:354 Described 346:354 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 2 9 theme sulfation 326:334 arg1 patterns 336:343 the various sulfation patterns 314:343 the various sulfation patterns 314:343 The synthesis of structurally diverse but defined GAGs is extremely challenging as one has to account for the various sulfation patterns. 25325938 1 10 theme important 91:99 arg1 carbohydrates 110:122 important sulfated carbohydrates 91:122 important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions 91:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 1 10 theme important 91:99 arg1 Glycosaminoglycans 61:78 Glycosaminoglycans 61:78 Glycosaminoglycans (GAGs) 61:85 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 6 11 theme GAG 743:745 arg1 families 747:754 all GAG families 739:754 all GAG families 739:754 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 25325938 0 12 theme Automated 0:8 arg1 synthesis 10:18 Automated synthesis 0:18 Automated synthesis of chondroitin sulfate 0:41 Automated synthesis of chondroitin sulfate oligosaccharides. 25325938 0 13 theme chondroitin 23:33 arg1 sulfate 35:41 chondroitin sulfate 23:41 chondroitin sulfate 23:41 Automated synthesis of chondroitin sulfate oligosaccharides. 25325938 5 14 theme mild 623:626 arg1 conditions 628:637 chemically mild conditions 612:637 chemically mild conditions 612:637 The linker cleavage from the resin is performed in a continuous-flow photoreactor under chemically mild conditions. 25325938 1 15 theme sulfated 101:108 arg1 carbohydrates 110:122 important sulfated carbohydrates 91:122 important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions 91:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 1 15 theme sulfated 101:108 arg1 Glycosaminoglycans 61:78 Glycosaminoglycans 61:78 Glycosaminoglycans (GAGs) 61:85 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 5 16 from resin 553:557 arg1 cleavage 535:542 The linker cleavage 524:542 The linker cleavage from the resin 524:557 The linker cleavage from the resin is performed in a continuous-flow photoreactor under chemically mild conditions. 25325938 3 17 theme chondroitin 390:400 arg1 hexasaccharides 410:424 two chondroitin sulfate hexasaccharides 386:424 two chondroitin sulfate hexasaccharides 386:424 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 2 18 theme various 318:324 arg1 patterns 336:343 the various sulfation patterns 314:343 the various sulfation patterns 314:343 The synthesis of structurally diverse but defined GAGs is extremely challenging as one has to account for the various sulfation patterns. 25325938 3 19 theme sulfate 402:408 arg1 hexasaccharides 410:424 two chondroitin sulfate hexasaccharides 386:424 two chondroitin sulfate hexasaccharides 386:424 Described is the automated synthesis of two chondroitin sulfate hexasaccharides. 25325938 0 20 theme sulfate 35:41 arg1 synthesis 10:18 Automated synthesis 0:18 Automated synthesis of chondroitin sulfate 0:41 Automated synthesis of chondroitin sulfate oligosaccharides. 25325938 2 21 theme GAGs 258:261 arg1 synthesis 212:220 The synthesis 208:220 The synthesis of structurally diverse but defined GAGs 208:261 The synthesis of structurally diverse but defined GAGs is extremely challenging as one has to account for the various sulfation patterns. 25325938 2 22 theme defined 250:256 arg1 GAGs 258:261 structurally diverse but defined GAGs 225:261 structurally diverse but defined GAGs 225:261 The synthesis of structurally diverse but defined GAGs is extremely challenging as one has to account for the various sulfation patterns. 25325938 2 23 theme diverse 238:244 arg1 GAGs 258:261 structurally diverse but defined GAGs 225:261 structurally diverse but defined GAGs 225:261 The synthesis of structurally diverse but defined GAGs is extremely challenging as one has to account for the various sulfation patterns. 25325938 5 24 theme continuous-flow 577:591 arg1 photoreactor 593:604 a continuous-flow photoreactor 575:604 a continuous-flow photoreactor 575:604 The linker cleavage from the resin is performed in a continuous-flow photoreactor under chemically mild conditions. 25325938 1 25 theme extracellular 149:161 arg1 matrix 163:168 the extracellular matrix 145:168 the extracellular matrix that serve many biological functions 145:205 Glycosaminoglycans (GAGs) are important sulfated carbohydrates prevalently found in the extracellular matrix that serve many biological functions. 25325938 4 26 theme solid 466:470 arg1 support 472:478 a solid support 464:478 a solid support that is equipped with a photolabile linker 464:521 The oligosaccharides are prepared on a solid support that is equipped with a photolabile linker. 25325938 6 27 theme general 679:685 arg1 scheme 687:692 a general scheme 677:692 a general scheme 677:692 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 25325938 6 27 theme general 679:685 arg1 approach 654:661 The described approach 640:661 The described approach 640:661 The described approach will serve as a general scheme to systematically access oligosaccharides of all GAG families. 28461410 4 0 theme 34-103 819:824 arg1 structures 835:844 34-103 O-glycan structures 819:844 34-103 O-glycan structures 819:844 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 9 1 from tissue 1638:1643 arg1 O-glycans 1613:1621 gastric O-glycans 1605:1621 gastric O-glycans from cancerous tissue than from healthy stomachs 1605:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 8 2 theme gastric 1311:1317 arg1 O-glycome 1319:1327 the human gastric O-glycome 1301:1327 the human gastric O-glycome 1301:1327 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 6 3 theme Blood 960:964 arg1 antigens 974:981 Blood group I antigens 960:981 Blood group I antigens 960:981 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 1 4 gly O-glycosylation 63:77 arg1 individuals 85:95 10 individuals 82:95 10 individuals 82:95 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 1 4 gly O-glycosylation 63:77 arg1 depth 146:150 depth 146:150 depth 146:150 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 4 5 from individual 730:739 arg1 mucins 709:714 mucins 709:714 mucins from a single individual 709:739 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 8 6 theme human 1305:1309 arg1 O-glycome 1319:1327 the human gastric O-glycome 1301:1327 the human gastric O-glycome 1301:1327 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 4 7 contain carried 811:817 arg2 structures 835:844 34-103 O-glycan structures 819:844 34-103 O-glycan structures 819:844 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 4 7 contain carried 811:817 arg1 individual 800:809 each individual 795:809 each individual 795:809 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 9 8 theme cancerous 1628:1636 arg1 tissue 1638:1643 cancerous tissue 1628:1643 cancerous tissue 1628:1643 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 8 9 theme cancer 1362:1367 arg1 research 1369:1376 gastric cancer research 1354:1376 gastric cancer research 1354:1376 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 9 10 theme gastric 1605:1611 arg1 O-glycans 1613:1621 gastric O-glycans 1605:1621 gastric O-glycans from cancerous tissue than from healthy stomachs 1605:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 5 11 theme gastric 912:918 arg1 O-glycans 920:928 gastric O-glycans 912:928 gastric O-glycans 912:928 The majority of gastric O-glycans were neutral and fucosylated. 28461410 4 12 theme enormous 613:620 arg1 diversity 622:630 An enormous diversity 610:630 An enormous diversity in glycosylation 610:647 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 8 13 dep diversity 1219:1227 arg1 The 1215:1217 The 1215:1217 The 1215:1217 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 4 14 theme O-glycan 826:833 arg1 structures 835:844 34-103 O-glycan structures 819:844 34-103 O-glycan structures 819:844 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 1 15 theme mucin 57:61 arg1 O-glycosylation 63:77 The mucin O-glycosylation 53:77 The mucin O-glycosylation of 10 individuals with and without gastric disease 53:128 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 2 16 theme gastric 485:491 arg1 cancer 493:498 gastric cancer 485:498 gastric cancer 485:498 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 6 17 theme human 1093:1097 arg1 O-glycans 1107:1115 human gastric O-glycans 1093:1115 human gastric O-glycans 1093:1115 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 9 18 theme healthy 1655:1661 arg1 stomachs 1663:1670 healthy stomachs 1655:1670 healthy stomachs 1655:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 4 19 from diversity 622:630 arg1 glycosylation 635:647 glycosylation 635:647 glycosylation 635:647 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 2 20 theme interaction 369:379 arg1 point 360:364 the first point 350:364 the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer 350:498 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 7 21 theme glycan 1159:1164 arg1 structures 1166:1175 1-14 glycan structures 1154:1175 1-14 glycan structures that were unique for that individual 1154:1212 Furthemore, each individual carried 1-14 glycan structures that were unique for that individual. 28461410 4 22 theme single 723:728 arg1 individual 730:739 a single individual 721:739 a single individual 721:739 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 6 23 dep α1,4-GlcNAc-like 1004:1019 arg1 LacdiNAc-like 1048:1060 LacdiNAc-like 1048:1060 LacdiNAc-like 1048:1060 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 2 24 theme gastric 453:459 arg1 ulcers 474:479 gastric and duodenal ulcers 453:479 gastric and duodenal ulcers 453:479 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 2 24 theme gastric 453:459 arg1 pylori 416:421 Helicobacter pylori 403:421 Helicobacter pylori 403:421 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 2 25 from juice 332:336 arg1 surface 300:306 the epithelial surface 285:306 the epithelial surface from the acidic gastric juice 285:336 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 8 26 theme gastric 1354:1360 arg1 research 1369:1376 gastric cancer research 1354:1376 gastric cancer research 1354:1376 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 0 27 theme Structural 0:9 arg1 Diversity 11:19 Structural Diversity 0:19 Structural Diversity of Human Gastric Mucin Glycans 0:50 Structural Diversity of Human Gastric Mucin Glycans. 28461410 9 28 theme higher 1560:1565 arg1 level 1567:1571 a higher level 1558:1571 a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs 1558:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 7 29 contain carried 1146:1152 arg1 individual 1135:1144 each individual 1130:1144 each individual 1130:1144 Furthemore, each individual carried 1-14 glycan structures that were unique for that individual. 28461410 7 29 contain carried 1146:1152 arg2 structures 1166:1175 1-14 glycan structures 1154:1175 1-14 glycan structures that were unique for that individual 1154:1212 Furthemore, each individual carried 1-14 glycan structures that were unique for that individual. 28461410 2 30 theme epithelial 289:298 arg1 surface 300:306 the epithelial surface 285:306 the epithelial surface from the acidic gastric juice 285:336 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 2 31 theme first 354:358 arg1 point 360:364 the first point 350:364 the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer 350:498 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 0 32 theme Human 24:28 arg1 Glycans 44:50 Human Gastric Mucin Glycans 24:50 Human Gastric Mucin Glycans 24:50 Structural Diversity of Human Gastric Mucin Glycans. 28461410 9 33 theme sulfation 1592:1600 arg1 level 1567:1571 a higher level 1558:1571 a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs 1558:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 3 34 theme present 521:527 arg1 study 529:533 the present study 517:533 the present study 517:533 The rational of the present study was to map the O-glycosylation that the pathogen may come in contact with. 28461410 1 35 theme structural 175:184 arg1 map 186:188 a structural map 173:188 a structural map of human gastric glycosylation 173:219 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 1 36 theme individuals 85:95 arg1 O-glycosylation 63:77 The mucin O-glycosylation 53:77 The mucin O-glycosylation of 10 individuals with and without gastric disease 53:128 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 3 37 theme study 529:533 arg1 rational 505:512 rational 505:512 rational 505:512 The rational of the present study was to map the O-glycosylation that the pathogen may come in contact with. 28461410 0 38 theme Mucin 38:42 arg1 Glycans 44:50 Human Gastric Mucin Glycans 24:50 Human Gastric Mucin Glycans 24:50 Structural Diversity of Human Gastric Mucin Glycans. 28461410 2 39 theme acidic 317:322 arg1 juice 332:336 the acidic gastric juice 313:336 the acidic gastric juice 313:336 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 7 40 theme 1-14 1154:1157 arg1 structures 1166:1175 1-14 glycan structures 1154:1175 1-14 glycan structures that were unique for that individual 1154:1212 Furthemore, each individual carried 1-14 glycan structures that were unique for that individual. 28461410 8 41 theme gastric 1248:1254 arg1 O-glycosylation 1256:1270 gastric O-glycosylation 1248:1270 gastric O-glycosylation 1248:1270 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 0 42 theme Gastric 30:36 arg1 Glycans 44:50 Human Gastric Mucin Glycans 24:50 Human Gastric Mucin Glycans 24:50 Structural Diversity of Human Gastric Mucin Glycans. 28461410 9 43 theme sialylation 1576:1586 arg1 level 1567:1571 a higher level 1558:1571 a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs 1558:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 8 44 from diversity 1219:1227 arg1 O-glycosylation 1256:1270 gastric O-glycosylation 1248:1270 gastric O-glycosylation 1248:1270 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 1 45 theme human 193:197 arg1 glycosylation 207:219 human gastric glycosylation 193:219 human gastric glycosylation 193:219 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 9 46 from stomachs 1663:1670 arg1 O-glycans 1613:1621 gastric O-glycans 1605:1621 gastric O-glycans from cancerous tissue than from healthy stomachs 1605:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 5 47 theme O-glycans 920:928 arg1 majority 900:907 The majority 896:907 The majority of gastric O-glycans 896:928 The majority of gastric O-glycans were neutral and fucosylated. 28461410 5 47 theme O-glycans 920:928 arg1 neutral 935:941 neutral 935:941 neutral 935:941 The majority of gastric O-glycans were neutral and fucosylated. 28461410 2 48 theme gastric 324:330 arg1 juice 332:336 the acidic gastric juice 313:336 the acidic gastric juice 313:336 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 0 49 theme Glycans 44:50 arg1 Diversity 11:19 Structural Diversity 0:19 Structural Diversity of Human Gastric Mucin Glycans 0:50 Structural Diversity of Human Gastric Mucin Glycans. 28461410 8 50 theme cancer 1466:1471 arg1 structures 1482:1491 gastric cancer specific structures 1458:1491 gastric cancer specific structures 1458:1491 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 8 51 theme gastric 1458:1464 arg1 structures 1482:1491 gastric cancer specific structures 1458:1491 gastric cancer specific structures 1458:1491 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 1 52 theme gastric 114:120 arg1 disease 122:128 gastric disease 114:128 gastric disease 114:128 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 4 53 theme glycan 748:753 arg1 chain 755:759 mucin glycan chain 742:759 mucin glycan chain length 742:766 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 8 54 theme individual 1406:1415 arg1 variation 1417:1425 the high individual variation 1397:1425 the high individual variation 1397:1425 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 4 55 theme mucin 742:746 arg1 chain 755:759 mucin glycan chain 742:759 mucin glycan chain length 742:766 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 9 56 theme low 1515:1517 arg1 number 1519:1524 the low number 1511:1524 the low number of individuals 1511:1539 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 6 57 theme O-glycans 1107:1115 arg1 modifications 1076:1088 common modifications 1069:1088 common modifications of human gastric O-glycans 1069:1115 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 1 58 theme gastric 199:205 arg1 glycosylation 207:219 human gastric glycosylation 193:219 human gastric glycosylation 193:219 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 9 59 theme individuals 1529:1539 arg1 number 1519:1524 the low number 1511:1524 the low number of individuals 1511:1539 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 8 60 theme high 1401:1404 arg1 variation 1417:1425 the high individual variation 1397:1425 the high individual variation 1397:1425 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 6 61 theme common 1069:1074 arg1 modifications 1076:1088 common modifications 1069:1088 common modifications of human gastric O-glycans 1069:1115 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 8 62 theme O-glycome 1319:1327 arg1 understanding 1284:1296 our understanding 1280:1296 our understanding of the human gastric O-glycome 1280:1327 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 8 62 theme O-glycome 1319:1327 arg1 implications 1337:1348 its implications 1333:1348 its implications for gastric cancer research 1333:1376 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 1 63 theme glycosylation 207:219 arg1 map 186:188 a structural map 173:188 a structural map of human gastric glycosylation 173:219 The mucin O-glycosylation of 10 individuals with and without gastric disease was examined in depth in order to generate a structural map of human gastric glycosylation. 28461410 8 64 theme specific 1473:1480 arg1 structures 1482:1491 gastric cancer specific structures 1458:1491 gastric cancer specific structures 1458:1491 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 4 65 theme chain 755:759 arg1 length 761:766 mucin glycan chain length 742:766 mucin glycan chain length 742:766 An enormous diversity in glycosylation was found, which varied both between individuals and within mucins from a single individual: mucin glycan chain length ranged from 2-13 residues, each individual carried 34-103 O-glycan structures and in total over 258 structures were identified. 28461410 9 66 from level 1567:1571 arg1 O-glycans 1613:1621 gastric O-glycans 1605:1621 gastric O-glycans from cancerous tissue than from healthy stomachs 1605:1670 However, despite the low number of individuals, we could verify a higher level of sialylation and sulfation on gastric O-glycans from cancerous tissue than from healthy stomachs. 28461410 2 67 theme duodenal 465:472 arg1 ulcers 474:479 gastric and duodenal ulcers 453:479 gastric and duodenal ulcers 453:479 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 2 67 theme duodenal 465:472 arg1 pylori 416:421 Helicobacter pylori 403:421 Helicobacter pylori 403:421 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 6 68 theme gastric 1099:1105 arg1 O-glycans 1107:1115 human gastric O-glycans 1093:1115 human gastric O-glycans 1093:1115 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 28461410 2 69 theme Helicobacter 403:414 arg1 pylori 416:421 Helicobacter pylori 403:421 Helicobacter pylori 403:421 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 8 70 from alterations 1233:1243 arg1 O-glycosylation 1256:1270 gastric O-glycosylation 1248:1270 gastric O-glycosylation 1248:1270 The diversity and alterations in gastric O-glycosylation broaden our understanding of the human gastric O-glycome and its implications for gastric cancer research and emphasize that the high individual variation makes it difficult to identify gastric cancer specific structures. 28461410 2 71 dep mucins 244:249 arg1 these 238:242 these 238:242 these 238:242 In the stomach, these mucins and their O-glycosylation protect the epithelial surface from the acidic gastric juice and provide the first point of interaction for pathogens such as Helicobacter pylori, reported to cause gastritis, gastric and duodenal ulcers and gastric cancer. 28461410 6 72 theme group 966:970 arg1 antigens 974:981 Blood group I antigens 960:981 Blood group I antigens 960:981 Blood group I antigens, as well as terminal α1,4-GlcNAc-like and GalNAcβ1-4GlcNAc-like (LacdiNAc-like), were common modifications of human gastric O-glycans. 24571125 0 0 theme β-KDO-containing 93:108 arg1 oligosaccharides 110:125 β-KDO-containing oligosaccharides 93:125 β-KDO-containing oligosaccharides 93:125 Preparation of a protected 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides. 24571125 1 1 theme glycal 172:177 arg1 donors 179:184 KDO glycal donors 168:184 KDO glycal donors 168:184 A practical method for the synthesis of KDO glycal donors was developed. 24571125 1 2 theme donors 179:184 arg1 synthesis 155:163 the synthesis 151:163 the synthesis of KDO glycal donors 151:184 A practical method for the synthesis of KDO glycal donors was developed. 24571125 0 3 theme oligosaccharides 110:125 arg1 synthesis 80:88 the synthesis 76:88 the synthesis of β-KDO-containing oligosaccharides 76:125 Preparation of a protected 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides. 24571125 3 4 theme oligosaccharides 508:523 arg1 preparation 476:486 the preparation 472:486 the preparation of β-KDO-containing oligosaccharides 472:523 The synthetic use of the KDO donor was demonstrated in the preparation of β-KDO-containing oligosaccharides. 24571125 3 5 theme KDO 442:444 arg1 donor 446:450 the KDO donor 438:450 the KDO donor 438:450 The synthetic use of the KDO donor was demonstrated in the preparation of β-KDO-containing oligosaccharides. 24571125 3 6 theme donor 446:450 arg1 use 431:433 The synthetic use 417:433 The synthetic use of the KDO donor 417:450 The synthetic use of the KDO donor was demonstrated in the preparation of β-KDO-containing oligosaccharides. 24571125 0 7 theme a 15:15 arg1 Preparation 0:10 Preparation 0:10 Preparation of a 0:15 Preparation of a protected 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides. 24571125 1 8 theme practical 130:138 arg1 method 140:145 A practical method 128:145 A practical method for the synthesis of KDO glycal donors 128:184 A practical method for the synthesis of KDO glycal donors was developed. 24571125 0 9 theme glycal 59:64 arg1 donor 66:70 3-deoxy-D-manno-oct-2-ulosonate glycal donor 27:70 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides 27:125 Preparation of a protected 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides. 24571125 2 10 theme glycosylation 270:282 arg1 disastereoselectivity 245:265 excellent disastereoselectivity 235:265 excellent disastereoselectivity of glycosylation 235:282 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 2 11 theme C-5 402:404 arg1 hydroxyls 406:414 the C-4 and C-5 hydroxyls 390:414 hydroxyls 406:414 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 0 12 theme 3-deoxy-D-manno-oct-2-ulosonate 27:57 arg1 donor 66:70 3-deoxy-D-manno-oct-2-ulosonate glycal donor 27:70 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides 27:125 Preparation of a protected 3-deoxy-D-manno-oct-2-ulosonate glycal donor for the synthesis of β-KDO-containing oligosaccharides. 24571125 2 13 from hydroxyls 406:414 arg1 protection 376:385 the isopropylidene protection 357:385 the isopropylidene protection at the C-4 and C-5 hydroxyls 357:414 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 3 14 theme synthetic 421:429 arg1 use 431:433 The synthetic use 417:433 The synthetic use of the KDO donor 417:450 The synthetic use of the KDO donor was demonstrated in the preparation of β-KDO-containing oligosaccharides. 24571125 2 15 theme excellent 235:243 arg1 disastereoselectivity 245:265 excellent disastereoselectivity 235:265 excellent disastereoselectivity of glycosylation 235:282 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 2 16 theme isopropylidene 361:374 arg1 protection 376:385 the isopropylidene protection 357:385 the isopropylidene protection at the C-4 and C-5 hydroxyls 357:414 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 2 17 theme solvent 302:308 arg1 mixture 310:316 a CH2Cl2-CH3CN solvent mixture 287:316 a CH2Cl2-CH3CN solvent mixture 287:316 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 3 18 theme β-KDO-containing 491:506 arg1 oligosaccharides 508:523 β-KDO-containing oligosaccharides 491:523 β-KDO-containing oligosaccharides 491:523 The synthetic use of the KDO donor was demonstrated in the preparation of β-KDO-containing oligosaccharides. 24571125 2 19 theme KDO 214:216 arg1 donors 218:223 The prepared KDO donors 201:223 The prepared KDO donors 201:223 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 2 20 theme CH2Cl2-CH3CN 289:300 arg1 mixture 310:316 a CH2Cl2-CH3CN solvent mixture 287:316 a CH2Cl2-CH3CN solvent mixture 287:316 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 1 21 theme KDO 168:170 arg1 donors 179:184 KDO glycal donors 168:184 KDO glycal donors 168:184 A practical method for the synthesis of KDO glycal donors was developed. 24571125 2 22 theme prepared 205:212 arg1 donors 218:223 The prepared KDO donors 201:223 The prepared KDO donors 201:223 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 24571125 2 23 theme C-4 394:396 arg1 hydroxyls 406:414 the C-4 and C-5 hydroxyls 390:414 hydroxyls 406:414 The prepared KDO donors exhibited excellent disastereoselectivity of glycosylation in a CH2Cl2-CH3CN solvent mixture, which was found to be associated with the isopropylidene protection at the C-4 and C-5 hydroxyls. 29096187 4 0 theme biotinylation 642:654 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 1 from removal 537:543 arg1 procedures 620:629 deprotection procedures 607:629 deprotection procedures 607:629 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 1 from removal 537:543 arg1 sulfation 592:600 sulfation 592:600 sulfation 592:600 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 1 from removal 537:543 arg1 O-6 572:574 O-6 572:574 O-6 of two GlcNAcs 572:589 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 2 theme GlcNAcs 583:589 arg1 procedures 620:629 deprotection procedures 607:629 deprotection procedures 607:629 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 2 theme GlcNAcs 583:589 arg1 sulfation 592:600 sulfation 592:600 sulfation 592:600 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 2 theme GlcNAcs 583:589 arg1 O-6 572:574 O-6 572:574 O-6 of two GlcNAcs 572:589 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 3 3 from Optimization 341:352 arg1 coupling 363:370 2 + 2 coupling 357:370 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature 357:434 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 4 4 theme protecting 552:561 arg1 group 563:567 the protecting group 548:567 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 5 from O-6 572:574 arg1 group 563:567 the protecting group 548:567 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 5 from O-6 572:574 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 5 from O-6 572:574 arg1 biotinylation 642:654 biotinylation 642:654 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 1 6 theme stereocontrolled 217:232 arg1 manner 234:239 a stereocontrolled manner 215:239 a stereocontrolled manner 215:239 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 4 7 theme group 563:567 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 3 8 theme glycal 489:494 arg1 formation 496:504 glycal formation 489:504 glycal formation 489:504 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 3 9 theme low 420:422 arg1 temperature 424:434 a low temperature 418:434 a low temperature 418:434 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 4 10 theme deprotection 607:618 arg1 procedures 620:629 deprotection procedures 607:629 deprotection procedures 607:629 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 3 11 theme 2 + 2 357:361 arg1 coupling 363:370 2 + 2 coupling 357:370 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature 357:434 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 4 12 theme chemoselective 522:535 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 0 13 theme sulfate 36:42 arg1 tetrasaccharide 44:58 a biotinylated keratan sulfate tetrasaccharide 13:58 a biotinylated keratan sulfate tetrasaccharide 13:58 Synthesis of a biotinylated keratan sulfate tetrasaccharide composed of dimeric Galβ1-4GlcNAc6Sβ. 29096187 4 14 theme subsequent 511:520 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 0 15 theme keratan 28:34 arg1 tetrasaccharide 44:58 a biotinylated keratan sulfate tetrasaccharide 13:58 a biotinylated keratan sulfate tetrasaccharide 13:58 Synthesis of a biotinylated keratan sulfate tetrasaccharide composed of dimeric Galβ1-4GlcNAc6Sβ. 29096187 0 16 theme biotinylated 15:26 arg1 tetrasaccharide 44:58 a biotinylated keratan sulfate tetrasaccharide 13:58 a biotinylated keratan sulfate tetrasaccharide 13:58 Synthesis of a biotinylated keratan sulfate tetrasaccharide composed of dimeric Galβ1-4GlcNAc6Sβ. 29096187 4 17 from procedures 620:629 arg1 group 563:567 the protecting group 548:567 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 17 from procedures 620:629 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 17 from procedures 620:629 arg1 biotinylation 642:654 biotinylation 642:654 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 3 18 theme desired 449:455 arg1 tetrasaccharide 457:471 the desired tetrasaccharide 445:471 the desired tetrasaccharide that suppressed glycal formation 445:504 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 1 19 theme biotinylated 130:141 arg1 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 1 19 theme biotinylated 130:141 arg1 tetrasaccharide 159:173 the biotinylated keratan sulfate tetrasaccharide 126:173 the biotinylated keratan sulfate tetrasaccharide 126:173 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 0 20 theme tetrasaccharide 44:58 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of a biotinylated keratan sulfate tetrasaccharide 0:58 Synthesis of a biotinylated keratan sulfate tetrasaccharide composed of dimeric Galβ1-4GlcNAc6Sβ. 29096187 3 21 from CuBr2 385:389 arg1 CH3NO2 408:413 CH3NO2 408:413 CH3NO2 408:413 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 1 22 theme keratan 143:149 arg1 sulfate 151:157 keratan sulfate 143:157 the biotinylated keratan sulfate tetrasaccharide 126:173 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 2 23 theme Galβ1-4GlcNPhth 265:279 arg1 unit 281:284 The suitably protected Galβ1-4GlcNPhth unit 242:284 The suitably protected Galβ1-4GlcNPhth unit 242:284 The suitably protected Galβ1-4GlcNPhth unit was converted to the corresponding donor and acceptor. 29096187 1 24 theme sulfate 151:157 arg1 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ 176:210 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 1 24 theme sulfate 151:157 arg1 tetrasaccharide 159:173 the biotinylated keratan sulfate tetrasaccharide 126:173 the biotinylated keratan sulfate tetrasaccharide 126:173 We successfully synthesized the biotinylated keratan sulfate tetrasaccharide, Galβ1-4GlcNAc6Sβ1-3Galβ1-4GlcNAc6Sβ in a stereocontrolled manner. 29096187 2 25 theme protected 255:263 arg1 unit 281:284 The suitably protected Galβ1-4GlcNPhth unit 242:284 The suitably protected Galβ1-4GlcNPhth unit 242:284 The suitably protected Galβ1-4GlcNPhth unit was converted to the corresponding donor and acceptor. 29096187 4 26 theme target 665:670 arg1 compound 672:679 the target compound 661:679 the target compound 661:679 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 0 27 theme dimeric 72:78 arg1 Galβ1-4GlcNAc6Sβ 80:95 dimeric Galβ1-4GlcNAc6Sβ 72:95 dimeric Galβ1-4GlcNAc6Sβ 72:95 Synthesis of a biotinylated keratan sulfate tetrasaccharide composed of dimeric Galβ1-4GlcNAc6Sβ. 29096187 3 28 from n-Bu4NBr 396:403 arg1 CH3NO2 408:413 CH3NO2 408:413 CH3NO2 408:413 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 3 29 from AgOTf 378:382 arg1 CH3NO2 408:413 CH3NO2 408:413 CH3NO2 408:413 Optimization in 2 + 2 coupling using AgOTf, CuBr2, and n-Bu4NBr in CH3NO2 at a low temperature afforded the desired tetrasaccharide that suppressed glycal formation. 29096187 4 30 from sulfation 592:600 arg1 group 563:567 the protecting group 548:567 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 30 from sulfation 592:600 arg1 removal 537:543 The subsequent chemoselective removal 507:543 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 507:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 4 30 from sulfation 592:600 arg1 biotinylation 642:654 biotinylation 642:654 the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation 548:654 The subsequent chemoselective removal of the protecting group at O-6 of two GlcNAcs, sulfation, and deprotection procedures as well as biotinylation gave the target compound. 29096187 2 31 theme corresponding 307:319 arg1 donor 321:325 the corresponding donor 303:325 the corresponding donor 303:325 The suitably protected Galβ1-4GlcNPhth unit was converted to the corresponding donor and acceptor. 26891276 5 0 gly glycosylated 973:984 arg1 capable 999:1005 capable 999:1005 capable 999:1005 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 5 0 gly glycosylated 973:984 arg1 peptides 986:993 the glycosylated peptides 969:993 the glycosylated peptides 969:993 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 4 1 theme glycosylated 607:618 arg1 variants 620:627 β-O-linked glycosylated variants 596:627 β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model 596:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 5 2 theme scaffold 1075:1082 arg1 aggregation 1021:1031 aggregation 1021:1031 aggregation of the non-modified corresponding amyloid scaffold 1021:1082 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 4 3 link Tau-derived 651:661 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 3 link Tau-derived 651:661 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 3 4 theme glycans 372:378 arg1 role 364:367 the role 360:367 the role of glycans in protein aggregation 360:401 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 2 5 theme many 315:318 arg1 diseases 330:337 many prevalent diseases 315:337 many prevalent diseases 315:337 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 0 6 theme Glycosylated 84:95 arg1 Analogues 97:105 Its Glycosylated Analogues 80:105 Its Glycosylated Analogues 80:105 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 4 7 theme hexapeptide 663:673 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 7 theme hexapeptide 663:673 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 3 8 theme misfolding 488:497 arg1 diseases 499:506 protein misfolding diseases 480:506 protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes 480:587 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 5 9 theme first 904:908 arg1 time 910:913 the first time 900:913 the first time 900:913 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 5 10 dep hinder 940:945 arg1 only 924:927 only 924:927 only 924:927 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 5 11 theme glycosylated 973:984 arg1 capable 999:1005 capable 999:1005 capable 999:1005 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 5 11 theme glycosylated 973:984 arg1 peptides 986:993 the glycosylated peptides 969:993 the glycosylated peptides 969:993 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 4 12 gly glycosylated 607:618 arg1 variants 620:627 β-O-linked glycosylated variants 596:627 β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model 596:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 0 13 theme Selective 0:8 arg1 Inhibition 10:19 Selective Inhibition 0:19 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide 0:72 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 0 14 theme Aggregation 24:34 arg1 Inhibition 10:19 Selective Inhibition 0:19 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide 0:72 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 2 15 theme diseases 330:337 arg1 hallmark 303:310 the hallmark 299:310 the hallmark of many prevalent diseases 299:337 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 15 theme diseases 330:337 arg1 Misfolding 236:245 Misfolding 236:245 Misfolding of protein monomers 236:265 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 15 theme diseases 330:337 arg1 aggregation 283:293 their toxic aggregation 271:293 their toxic aggregation 271:293 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 3 16 theme protein 383:389 arg1 aggregation 391:401 protein aggregation 383:401 protein aggregation 383:401 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 2 17 theme prevalent 320:328 arg1 diseases 330:337 many prevalent diseases 315:337 many prevalent diseases 315:337 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 3 18 theme protein 480:486 arg1 diseases 499:506 protein misfolding diseases 480:506 protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes 480:587 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 3 19 from role 364:367 arg1 aggregation 391:401 protein aggregation 383:401 protein aggregation 383:401 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 1 20 theme Protein 108:114 arg1 modification 165:176 a ubiquitous post-translational modification 133:176 a ubiquitous post-translational modification that regulates the folding and function of many proteins 133:233 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 1 20 theme Protein 108:114 arg1 glycosylation 116:128 Protein glycosylation 108:128 Protein glycosylation 108:128 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 1 21 theme many 221:224 arg1 proteins 226:233 many proteins 221:233 many proteins 221:233 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 0 22 theme Toxicity 40:47 arg1 Inhibition 10:19 Selective Inhibition 0:19 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide 0:72 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 2 23 theme monomers 258:265 arg1 hallmark 303:310 the hallmark 299:310 the hallmark of many prevalent diseases 299:337 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 23 theme monomers 258:265 arg1 Misfolding 236:245 Misfolding 236:245 Misfolding of protein monomers 236:265 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 23 theme monomers 258:265 arg1 aggregation 283:293 their toxic aggregation 271:293 their toxic aggregation 271:293 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 1 24 theme proteins 226:233 arg1 function 209:216 function 209:216 function 209:216 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 1 24 theme proteins 226:233 arg1 folding 197:203 folding 197:203 folding 197:203 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 4 25 theme amyloid 718:724 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 25 theme amyloid 718:724 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 26 theme β-O-linked 596:605 arg1 variants 620:627 β-O-linked glycosylated variants 596:627 β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model 596:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 2 27 theme protein 250:256 arg1 monomers 258:265 protein monomers 250:265 protein monomers 250:265 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 4 28 theme VQIVYK 681:686 arg1 variants 620:627 β-O-linked glycosylated variants 596:627 β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model 596:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 29 theme glycan 860:865 arg1 nature 846:851 the nature 842:851 the nature of the glycan itself 842:872 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 5 30 theme non-modified 1040:1051 arg1 scaffold 1075:1082 the non-modified corresponding amyloid scaffold 1036:1082 the non-modified corresponding amyloid scaffold 1036:1082 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 1 31 dep folding 197:203 arg1 the 193:195 the 193:195 the 193:195 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 5 32 theme corresponding 1053:1065 arg1 scaffold 1075:1082 the non-modified corresponding amyloid scaffold 1036:1082 the non-modified corresponding amyloid scaffold 1036:1082 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 4 33 theme glycan 816:821 arg1 unit 823:826 a glycan unit 814:826 a glycan unit 814:826 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 3 34 theme diseases 499:506 arg1 pathology 467:475 the pathology 463:475 the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes 463:587 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 3 35 theme therapeutic 568:578 arg1 purposes 580:587 therapeutic purposes 568:587 therapeutic purposes 568:587 Thus, understanding the role of glycans in protein aggregation is highly important and could contribute both to unraveling the pathology of protein misfolding diseases as well as providing a means for modifying their course for therapeutic purposes. 26891276 5 36 theme amyloid 1067:1073 arg1 scaffold 1075:1082 the non-modified corresponding amyloid scaffold 1036:1082 the non-modified corresponding amyloid scaffold 1036:1082 Importantly, we show for the first time that not only do glycans hinder self-aggregation, but the glycosylated peptides are capable of inhibiting aggregation of the non-modified corresponding amyloid scaffold. 26891276 2 37 theme toxic 277:281 arg1 hallmark 303:310 the hallmark 299:310 the hallmark of many prevalent diseases 299:337 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 37 theme toxic 277:281 arg1 Misfolding 236:245 Misfolding 236:245 Misfolding of protein monomers 236:265 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 2 37 theme toxic 277:281 arg1 aggregation 283:293 their toxic aggregation 271:293 their toxic aggregation 271:293 Misfolding of protein monomers and their toxic aggregation are the hallmark of many prevalent diseases. 26891276 1 38 theme ubiquitous 135:144 arg1 modification 165:176 a ubiquitous post-translational modification 133:176 a ubiquitous post-translational modification that regulates the folding and function of many proteins 133:233 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 1 38 theme ubiquitous 135:144 arg1 glycosylation 116:128 Protein glycosylation 108:128 Protein glycosylation 108:128 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 4 39 theme amyloid 753:759 arg1 formation 761:769 amyloid formation 753:769 amyloid formation 753:769 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 0 40 theme Peptide 66:72 arg1 Toxicity 40:47 Toxicity 40:47 Toxicity 40:47 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 0 40 theme Peptide 66:72 arg1 Aggregation 24:34 Aggregation 24:34 Aggregation 24:34 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 4 41 link β-O-linked 596:605 arg1 variants 620:627 β-O-linked glycosylated variants 596:627 β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model 596:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 42 theme motif 675:679 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 42 theme motif 675:679 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 1 43 theme post-translational 146:163 arg1 modification 165:176 a ubiquitous post-translational modification 133:176 a ubiquitous post-translational modification that regulates the folding and function of many proteins 133:233 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 1 43 theme post-translational 146:163 arg1 glycosylation 116:128 Protein glycosylation 108:128 Protein glycosylation 108:128 Protein glycosylation is a ubiquitous post-translational modification that regulates the folding and function of many proteins. 26891276 4 44 theme Tau-derived 651:661 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 44 theme Tau-derived 651:661 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 0 45 theme Tau-Derived 54:64 arg1 Peptide 66:72 a Tau-Derived Peptide 52:72 a Tau-Derived Peptide 52:72 Selective Inhibition of Aggregation and Toxicity of a Tau-Derived Peptide using Its Glycosylated Analogues. 26891276 4 46 theme simplified 707:716 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 46 theme simplified 707:716 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 47 theme studied 643:649 arg1 VQIVYK 681:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 the highly studied Tau-derived hexapeptide motif VQIVYK 632:686 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 26891276 4 47 theme studied 643:649 arg1 model 726:730 a simplified amyloid model 705:730 a simplified amyloid model 705:730 Using β-O-linked glycosylated variants of the highly studied Tau-derived hexapeptide motif VQIVYK, which served as a simplified amyloid model, we demonstrate that amyloid formation and toxicity can be strongly attenuated by a glycan unit, depending on the nature of the glycan itself. 27842516 0 0 theme thermophile 88:98 arg1 Geobacillus 100:110 the Gram-positive thermophile Geobacillus 70:110 the Gram-positive thermophile Geobacillus 70:110 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 5 1 theme thermophilic 831:842 arg1 Geobacillus 850:860 the Gram-positive thermophilic genus Geobacillus 813:860 the Gram-positive thermophilic genus Geobacillus 813:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 7 2 theme modified 1203:1210 arg1 sugars 1212:1217 distinct and heavily modified sugars 1182:1217 distinct and heavily modified sugars 1182:1217 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 1 3 theme sugar 192:196 arg1 chains 198:203 sugar chains 192:203 sugar chains 192:203 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 4 theme life 270:273 arg1 domains 259:265 all three domains 249:265 all three domains of life 249:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 4 theme life 270:273 arg1 life 270:273 life 270:273 life 270:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 0 5 theme Gram-positive 74:86 arg1 Geobacillus 100:110 the Gram-positive thermophile Geobacillus 70:110 the Gram-positive thermophile Geobacillus 70:110 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 5 6 theme Geobacillus 850:860 arg1 strains 802:808 multiple strains 793:808 multiple strains of the Gram-positive thermophilic genus Geobacillus 793:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 1 7 theme chains 198:203 arg1 attachment 178:187 the post-translational attachment 155:187 the post-translational attachment of sugar chains to target proteins 155:222 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 2 8 theme main 327:330 arg1 protein 343:349 the main structural protein 323:349 the main structural protein of the flagellum 323:366 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 8 theme main 327:330 arg1 flagellin 312:320 flagellin 312:320 flagellin 312:320 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 0 9 theme Geobacillus 100:110 arg1 island 60:65 the flagellin glycosylation island 32:65 the flagellin glycosylation island of the Gram-positive thermophile Geobacillus 32:110 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 5 10 theme flagellin 761:769 arg1 loci 785:788 the flagellin glycosylation loci 757:788 the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus 757:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 3 11 theme surface 594:600 arg1 recognition 602:612 surface recognition 594:612 surface recognition 594:612 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 7 12 theme glycosylation 1041:1053 arg1 islands 1055:1061 The Geobacillus flagellin glycosylation islands 1015:1061 The Geobacillus flagellin glycosylation islands (FGIs) 1015:1068 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 7 12 theme glycosylation 1041:1053 arg1 FGIs 1064:1067 FGIs 1064:1067 FGIs 1064:1067 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 2 13 theme common 374:379 arg1 characteristic 381:394 a common characteristic 372:394 a common characteristic among many Gram-negative bacteria and Archaea 372:440 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 13 theme common 374:379 arg1 glycosylation 295:307 Post-translational glycosylation 276:307 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, 276:367 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 5 14 theme strains 802:808 arg1 loci 785:788 the flagellin glycosylation loci 757:788 the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus 757:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 1 15 attach attachment 178:187 arg1 proteins 215:222 target proteins 208:222 target proteins 208:222 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 15 attach attachment 178:187 arg2 chains 198:203 sugar chains 192:203 sugar chains 192:203 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 5 16 dep identified 867:876 arg1 Using 720:724 Using 720:724 Using comparative genomic approaches 720:755 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 2 17 theme flagellin 312:320 arg1 characteristic 381:394 a common characteristic 372:394 a common characteristic among many Gram-negative bacteria and Archaea 372:440 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 17 theme flagellin 312:320 arg1 glycosylation 295:307 Post-translational glycosylation 276:307 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, 276:367 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 9 18 theme members 1540:1546 arg1 lifestyles 1526:1535 the lifestyles 1522:1535 the lifestyles of members of this thermophilic genus 1522:1573 This suggests that flagellin glycosylation plays an important role in the lifestyles of members of this thermophilic genus. 27842516 8 19 theme glycosylation 1309:1321 arg1 islands 1323:1329 flagellin glycosylation islands 1299:1329 flagellin glycosylation islands 1299:1329 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 3 20 theme Several 443:449 arg1 functions 460:468 Several distinct functions 443:468 Several distinct functions 443:468 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 7 21 theme Geobacillus 1019:1029 arg1 islands 1055:1061 The Geobacillus flagellin glycosylation islands 1015:1061 The Geobacillus flagellin glycosylation islands (FGIs) 1015:1068 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 7 21 theme Geobacillus 1019:1029 arg1 FGIs 1064:1067 FGIs 1064:1067 FGIs 1064:1067 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 0 22 theme Comparative 0:10 arg1 analysis 20:27 Comparative genomic analysis 0:27 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 0:111 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 1 23 theme target 208:213 arg1 proteins 215:222 target proteins 208:222 target proteins 208:222 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 0 24 theme genomic 12:18 arg1 analysis 20:27 Comparative genomic analysis 0:27 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 0:111 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 5 25 theme genus 844:848 arg1 Geobacillus 850:860 the Gram-positive thermophilic genus Geobacillus 813:860 the Gram-positive thermophilic genus Geobacillus 813:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 8 26 dep CONCLUSIONS 1220:1230 arg1 showed 1265:1270 showed 1265:1270 showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable 1265:1449 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 7 27 theme flagellin 1031:1039 arg1 islands 1055:1061 The Geobacillus flagellin glycosylation islands 1015:1061 The Geobacillus flagellin glycosylation islands (FGIs) 1015:1068 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 7 27 theme flagellin 1031:1039 arg1 FGIs 1064:1067 FGIs 1064:1067 FGIs 1064:1067 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 8 28 dep not 1284:1286 arg1 universal 1288:1296 universal 1288:1296 universal 1288:1296 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 2 29 theme Gram-negative 407:419 arg1 bacteria 421:428 many Gram-negative bacteria 402:428 many Gram-negative bacteria 402:428 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 30 gly glycosylation 295:307 arg1 protein 343:349 the main structural protein 323:349 the main structural protein of the flagellum 323:366 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 30 gly glycosylation 295:307 arg1 flagellin 312:320 flagellin 312:320 flagellin 312:320 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 6 31 theme thirty-six 909:918 arg1 strains 929:935 thirty-six compared strains 909:935 thirty-six compared strains of the genus 909:948 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 8 32 theme comparative 1236:1246 arg1 analyses 1256:1263 Our comparative genomic analyses 1232:1263 Our comparative genomic analyses 1232:1263 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 2 33 theme many 402:405 arg1 bacteria 421:428 many Gram-negative bacteria 402:428 many Gram-negative bacteria 402:428 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 1 34 theme BACKGROUND 113:122 arg1 glycosylation 132:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 9 35 theme genus 1569:1573 arg1 members 1540:1546 members 1540:1546 members of this thermophilic genus 1540:1573 This suggests that flagellin glycosylation plays an important role in the lifestyles of members of this thermophilic genus. 27842516 8 36 theme genomic 1248:1254 arg1 analyses 1256:1263 Our comparative genomic analyses 1232:1263 Our comparative genomic analyses 1232:1263 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 1 37 gly glycosylation 132:144 arg1 domains 259:265 all three domains 249:265 all three domains of life 249:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 37 gly glycosylation 132:144 arg1 life 270:273 life 270:273 life 270:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 38 theme Protein 124:130 arg1 glycosylation 132:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 6 39 theme horizontal 991:1000 arg1 acquisition 1002:1012 horizontal acquisition 991:1012 horizontal acquisition 991:1012 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 8 40 theme flagellin 1413:1421 arg1 glycans 1423:1429 the encoded flagellin glycans 1401:1429 the encoded flagellin glycans 1401:1429 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 8 40 theme flagellin 1413:1421 arg1 variable 1442:1449 variable 1442:1449 variable 1442:1449 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 0 41 theme flagellin 36:44 arg1 island 60:65 the flagellin glycosylation island 32:65 the flagellin glycosylation island of the Gram-positive thermophile Geobacillus 32:110 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 8 42 theme Geobacillus 1380:1390 arg1 members 1359:1365 members 1359:1365 members of the genus Geobacillus 1359:1390 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 5 43 theme glycosylation 771:783 arg1 loci 785:788 the flagellin glycosylation loci 757:788 the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus 757:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 9 44 theme thermophilic 1556:1567 arg1 genus 1569:1573 this thermophilic genus 1551:1573 this thermophilic genus 1551:1573 This suggests that flagellin glycosylation plays an important role in the lifestyles of members of this thermophilic genus. 27842516 8 45 theme genus 1374:1378 arg1 Geobacillus 1380:1390 the genus Geobacillus 1370:1390 the genus Geobacillus 1370:1390 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 3 46 theme distinct 451:458 arg1 functions 460:468 Several distinct functions 443:468 Several distinct functions 443:468 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 6 47 theme genus 944:948 arg1 strains 929:935 thirty-six compared strains 909:935 thirty-six compared strains of the genus 909:948 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 7 48 theme distinct 1097:1104 arg1 glycans 1159:1165 highly variable glycans 1143:1165 highly variable glycans decorated with distinct and heavily modified sugars 1143:1217 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 7 48 theme distinct 1097:1104 arg1 types 1106:1110 five distinct types 1092:1110 five distinct types 1092:1110 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 3 49 theme flagellin 492:500 arg1 glycosylation 502:514 flagellin glycosylation 492:514 flagellin glycosylation 492:514 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 0 50 theme island 60:65 arg1 analysis 20:27 Comparative genomic analysis 0:27 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 0:111 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 8 51 theme encoded 1405:1411 arg1 glycans 1423:1429 the encoded flagellin glycans 1401:1429 the encoded flagellin glycans 1401:1429 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 8 51 theme encoded 1405:1411 arg1 variable 1442:1449 variable 1442:1449 variable 1442:1449 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 0 52 theme glycosylation 46:58 arg1 island 60:65 the flagellin glycosylation island 32:65 the flagellin glycosylation island of the Gram-positive thermophile Geobacillus 32:110 Comparative genomic analysis of the flagellin glycosylation island of the Gram-positive thermophile Geobacillus. 27842516 9 53 theme flagellin 1471:1479 arg1 glycosylation 1481:1493 flagellin glycosylation 1471:1493 flagellin glycosylation 1471:1493 This suggests that flagellin glycosylation plays an important role in the lifestyles of members of this thermophilic genus. 27842516 5 54 theme comparative 726:736 arg1 approaches 746:755 comparative genomic approaches 726:755 comparative genomic approaches 726:755 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 7 55 theme distinct 1182:1189 arg1 sugars 1212:1217 distinct and heavily modified sugars 1182:1217 distinct and heavily modified sugars 1182:1217 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 8 56 theme flagellin 1299:1307 arg1 islands 1323:1329 flagellin glycosylation islands 1299:1329 flagellin glycosylation islands 1299:1329 CONCLUSIONS Our comparative genomic analyses showed that, while not universal, flagellin glycosylation islands are relatively common among members of the genus Geobacillus and that the encoded flagellin glycans are highly variable. 27842516 5 57 theme genomic 738:744 arg1 approaches 746:755 comparative genomic approaches 726:755 comparative genomic approaches 726:755 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 6 58 theme strains 929:935 arg1 strains 929:935 thirty-six compared strains 909:935 thirty-six compared strains of the genus 909:948 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 6 58 theme strains 929:935 arg1 Eighteen 897:904 Eighteen 897:904 Eighteen 897:904 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 2 59 theme flagellum 358:366 arg1 protein 343:349 the main structural protein 323:349 the main structural protein of the flagellum 323:366 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 59 theme flagellum 358:366 arg1 flagellin 312:320 flagellin 312:320 flagellin 312:320 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 60 theme structural 332:341 arg1 protein 343:349 the main structural protein 323:349 the main structural protein of the flagellum 323:366 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 60 theme structural 332:341 arg1 flagellin 312:320 flagellin 312:320 flagellin 312:320 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 7 61 theme variable 1150:1157 arg1 glycans 1159:1165 highly variable glycans 1143:1165 highly variable glycans decorated with distinct and heavily modified sugars 1143:1217 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 7 61 theme variable 1150:1157 arg1 types 1106:1110 five distinct types 1092:1110 five distinct types 1092:1110 The Geobacillus flagellin glycosylation islands (FGIs) can be clustered into five distinct types, which are predicted to encode highly variable glycans decorated with distinct and heavily modified sugars. 27842516 6 62 theme compared 920:927 arg1 strains 929:935 thirty-six compared strains 909:935 thirty-six compared strains of the genus 909:948 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 2 63 theme Post-translational 276:293 arg1 characteristic 381:394 a common characteristic 372:394 a common characteristic among many Gram-negative bacteria and Archaea 372:440 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 2 63 theme Post-translational 276:293 arg1 glycosylation 295:307 Post-translational glycosylation 276:307 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, 276:367 Post-translational glycosylation of flagellin, the main structural protein of the flagellum, is a common characteristic among many Gram-negative bacteria and Archaea. 27842516 1 64 theme post-translational 159:176 arg1 attachment 178:187 the post-translational attachment 155:187 the post-translational attachment of sugar chains to target proteins 155:222 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 6 65 contain carry 950:954 arg1 strains 929:935 thirty-six compared strains 909:935 thirty-six compared strains of the genus 909:948 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 6 65 contain carry 950:954 arg2 loci 962:965 these loci 956:965 these loci 956:965 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 6 65 contain carry 950:954 arg1 Eighteen 897:904 Eighteen 897:904 Eighteen 897:904 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 1 66 located observed 237:244 arg1 domains 259:265 all three domains 249:265 all three domains of life 249:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 66 located observed 237:244 arg2 glycosylation 132:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation 113:144 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 1 66 located observed 237:244 arg1 life 270:273 life 270:273 life 270:273 BACKGROUND Protein glycosylation involves the post-translational attachment of sugar chains to target proteins and has been observed in all three domains of life. 27842516 3 67 theme flagellar 564:572 arg1 filament 574:581 the flagellar filament 560:581 the flagellar filament 560:581 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 4 68 theme Gram-positive 688:700 arg1 bacteria 702:709 Gram-positive bacteria 688:709 Gram-positive bacteria 688:709 However, little is known about this trait among Gram-positive bacteria. 27842516 9 69 theme important 1504:1512 arg1 role 1514:1517 an important role 1501:1517 an important role 1501:1517 This suggests that flagellin glycosylation plays an important role in the lifestyles of members of this thermophilic genus. 27842516 5 70 theme Gram-positive 817:829 arg1 Geobacillus 850:860 the Gram-positive thermophilic genus Geobacillus 813:860 the Gram-positive thermophilic genus Geobacillus 813:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 27842516 6 71 theme acquisition 1002:1012 arg1 evidence 979:986 evidence 979:986 evidence of horizontal acquisition 979:1012 Eighteen of thirty-six compared strains of the genus carry these loci, which show evidence of horizontal acquisition. 27842516 3 72 theme filament 574:581 arg1 recognition 602:612 surface recognition 594:612 surface recognition 594:612 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 3 72 theme filament 574:581 arg1 adhesion 615:622 adhesion 615:622 adhesion 615:622 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 3 72 theme filament 574:581 arg1 stabilisation 527:539 stabilisation 527:539 stabilisation 527:539 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 3 72 theme filament 574:581 arg1 maintenance 545:555 maintenance 545:555 maintenance 545:555 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 3 72 theme filament 574:581 arg1 motility 584:591 motility 584:591 motility 584:591 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 3 72 theme filament 574:581 arg1 virulence 629:637 virulence 629:637 virulence 629:637 Several distinct functions have been ascribed to flagellin glycosylation, including stabilisation and maintenance of the flagellar filament, motility, surface recognition, adhesion, and virulence. 27842516 5 73 theme multiple 793:800 arg1 strains 802:808 multiple strains 793:808 multiple strains of the Gram-positive thermophilic genus Geobacillus 793:860 RESULTS Using comparative genomic approaches the flagellin glycosylation loci of multiple strains of the Gram-positive thermophilic genus Geobacillus were identified and characterized. 26232512 0 0 theme glycans 87:93 arg1 expression 56:65 cell surface expression 43:65 cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells 43:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 1 1 theme pancreatic 177:186 arg1 tumors 188:193 pancreatic tumors 177:193 pancreatic tumors 177:193 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 0 2 theme α1,2-fucosylated 70:85 arg1 glycans 87:93 α1,2-fucosylated glycans 70:93 α1,2-fucosylated glycans 70:93 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 4 3 theme FUT1/2 736:741 arg1 expression 743:752 FUT1/2 expression 736:752 FUT1/2 expression 736:752 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 0 4 theme human 98:102 arg1 cells 130:134 human pancreatic adenocarcinoma cells 98:134 human pancreatic adenocarcinoma cells 98:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 3 5 theme structures 618:627 arg1 levels 556:561 elevated FUT1 and FUT2 transcript levels 522:561 elevated FUT1 and FUT2 transcript levels 522:561 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 5 theme structures 618:627 arg1 expression 580:589 an increased expression 567:589 an increased expression of α1,2-fucosylated glycan structures on the surface of these cells 567:657 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 6 theme increased 570:578 arg1 expression 580:589 an increased expression 567:589 an increased expression of α1,2-fucosylated glycan structures on the surface of these cells 567:657 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 1 7 from activity 165:172 arg1 tumors 188:193 pancreatic tumors 177:193 pancreatic tumors 177:193 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 2 8 dep lines 357:361 arg1 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 8 dep lines 357:361 arg1 lines 357:361 the pancreatic cancer cell lines 330:361 the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T 330:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 8 dep lines 357:361 arg1 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 4 9 from first 692:696 arg1 conclusion 663:672 conclusion 663:672 conclusion 663:672 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 0 10 theme adenocarcinoma 115:128 arg1 cells 130:134 human pancreatic adenocarcinoma cells 98:134 human pancreatic adenocarcinoma cells 98:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 2 11 theme α1,2-fucosyltransferase 428:450 arg1 FUT2 467:470 FUT2 467:470 FUT2 467:470 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 11 theme α1,2-fucosyltransferase 428:450 arg1 FUT1 458:461 FUT1 458:461 FUT1 458:461 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 11 theme α1,2-fucosyltransferase 428:450 arg1 genes 452:456 the α1,2-fucosyltransferase genes 424:456 the α1,2-fucosyltransferase genes FUT1 and FUT2 424:470 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 0 12 gly α1,2-fucosylated 70:85 arg1 glycans 87:93 α1,2-fucosylated glycans 70:93 α1,2-fucosylated glycans 70:93 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 0 13 theme pancreatic 104:113 arg1 cells 130:134 human pancreatic adenocarcinoma cells 98:134 human pancreatic adenocarcinoma cells 98:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 4 14 theme cell-surface 795:806 arg1 glycans 808:814 cell-surface glycans 795:814 cell-surface glycans 795:814 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 0 15 theme inducible 8:16 arg1 down 28:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 3 16 theme α1,2-fucosylated 594:609 arg1 structures 618:627 α1,2-fucosylated glycan structures 594:627 α1,2-fucosylated glycan structures 594:627 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 17 theme glycan 611:616 arg1 structures 618:627 α1,2-fucosylated glycan structures 594:627 α1,2-fucosylated glycan structures 594:627 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 0 18 theme 1α 25:26 arg1 down 28:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 4 19 theme expression 743:752 arg1 HIF-1α 710:715 HIF-1α 710:715 HIF-1α 710:715 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 4 19 theme expression 743:752 arg1 suppressor 722:731 a suppressor 720:731 a suppressor of FUT1/2 expression 720:752 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 2 20 theme genes 452:456 arg1 expression 410:419 the expression 406:419 the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2 406:470 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 0 21 theme factor 18:23 arg1 down 28:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down 0:31 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 4 22 theme glycans 808:814 arg1 α1,2-fucosylation 774:790 α1,2-fucosylation 774:790 α1,2-fucosylation of cell-surface glycans 774:814 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 3 23 theme elevated 522:529 arg1 levels 556:561 elevated FUT1 and FUT2 transcript levels 522:561 elevated FUT1 and FUT2 transcript levels 522:561 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 4 24 from conclusion 663:672 arg1 data 679:682 our data 675:682 our data 675:682 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 4 24 from conclusion 663:672 arg1 first 692:696 first 692:696 first 692:696 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 1 25 theme normal 221:226 arg1 tissue 239:244 normal pancreatic tissue 221:244 normal pancreatic tissue 221:244 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 3 26 theme FUT1 531:534 arg1 levels 556:561 elevated FUT1 and FUT2 transcript levels 522:561 elevated FUT1 and FUT2 transcript levels 522:561 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 27 gly α1,2-fucosylated 594:609 arg1 structures 618:627 α1,2-fucosylated glycan structures 594:627 α1,2-fucosylated glycan structures 594:627 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 28 theme Down 473:476 arg1 regulation 478:487 Down regulation 473:487 Down regulation of HIF-1α expression 473:508 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 1 29 theme pancreatic 228:237 arg1 tissue 239:244 normal pancreatic tissue 221:244 normal pancreatic tissue 221:244 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 0 30 theme surface 48:54 arg1 expression 56:65 cell surface expression 43:65 cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells 43:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 0 31 from expression 56:65 arg1 cells 130:134 human pancreatic adenocarcinoma cells 98:134 human pancreatic adenocarcinoma cells 98:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 2 32 dep genes 452:456 arg1 FUT2 467:470 FUT2 467:470 FUT2 467:470 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 32 dep genes 452:456 arg1 FUT1 458:461 FUT1 458:461 FUT1 458:461 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 32 dep genes 452:456 arg1 genes 452:456 the α1,2-fucosyltransferase genes 424:456 the α1,2-fucosyltransferase genes FUT1 and FUT2 424:470 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 33 theme cell 352:355 arg1 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 33 theme cell 352:355 arg1 lines 357:361 the pancreatic cancer cell lines 330:361 the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T 330:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 33 theme cell 352:355 arg1 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 0 34 theme cell 43:46 arg1 expression 56:65 cell surface expression 43:65 cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells 43:134 Hypoxia inducible factor 1α down regulates cell surface expression of α1,2-fucosylated glycans in human pancreatic adenocarcinoma cells. 26232512 2 35 theme cancer 345:350 arg1 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 35 theme cancer 345:350 arg1 lines 357:361 the pancreatic cancer cell lines 330:361 the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T 330:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 35 theme cancer 345:350 arg1 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 3 36 from expression 580:589 arg1 surface 636:642 the surface 632:642 the surface of these cells 632:657 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 37 theme cells 653:657 arg1 surface 636:642 the surface 632:642 the surface of these cells 632:657 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 2 38 theme pancreatic 334:343 arg1 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Pa-Tu-8988S 363:373 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 38 theme pancreatic 334:343 arg1 lines 357:361 the pancreatic cancer cell lines 330:361 the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T 330:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 38 theme pancreatic 334:343 arg1 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Pa-Tu-8988T 379:389 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 39 theme inducible 273:281 arg1 HIF 291:293 HIF 291:293 HIF 291:293 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 2 39 theme inducible 273:281 arg1 factor 283:288 hypoxia inducible factor 265:288 hypoxia inducible factor (HIF) 265:294 Here we show that hypoxia inducible factor (HIF) 1α is constitutively expressed in the pancreatic cancer cell lines Pa-Tu-8988S and Pa-Tu-8988T and suppresses the expression of the α1,2-fucosyltransferase genes FUT1 and FUT2. 26232512 1 40 theme α1,2-fucosyltransferase 141:163 arg1 lower 203:207 lower 203:207 lower 203:207 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 1 40 theme α1,2-fucosyltransferase 141:163 arg1 activity 165:172 The α1,2-fucosyltransferase activity 137:172 The α1,2-fucosyltransferase activity in pancreatic tumors 137:193 The α1,2-fucosyltransferase activity in pancreatic tumors is much lower compared to normal pancreatic tissue. 26232512 3 41 theme HIF-1α 492:497 arg1 expression 499:508 HIF-1α expression 492:508 HIF-1α expression 492:508 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 42 theme expression 499:508 arg1 regulation 478:487 Down regulation 473:487 Down regulation of HIF-1α expression 473:508 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 4 43 gly α1,2-fucosylation 774:790 arg1 glycans 808:814 cell-surface glycans 795:814 cell-surface glycans 795:814 In conclusion, our data are the first to identify HIF-1α as a suppressor of FUT1/2 expression, thereby regulating α1,2-fucosylation of cell-surface glycans. 26232512 3 44 theme FUT2 540:543 arg1 levels 556:561 elevated FUT1 and FUT2 transcript levels 522:561 elevated FUT1 and FUT2 transcript levels 522:561 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 45 theme transcript 545:554 arg1 levels 556:561 elevated FUT1 and FUT2 transcript levels 522:561 elevated FUT1 and FUT2 transcript levels 522:561 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26232512 3 46 from levels 556:561 arg1 surface 636:642 the surface 632:642 the surface of these cells 632:657 Down regulation of HIF-1α expression resulted in elevated FUT1 and FUT2 transcript levels and an increased expression of α1,2-fucosylated glycan structures on the surface of these cells. 26597529 6 0 theme Vmax 938:941 arg1 values 952:957 Vmax and kcat values 938:957 values 952:957 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 9 1 dep P. 1316:1317 arg1 pastoris 1319:1326 pastoris 1319:1326 pastoris 1319:1326 This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA. 26597529 0 2 from Expression 13:22 arg1 oryzae 104:109 Aspergillus oryzae 92:109 Aspergillus oryzae 92:109 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 9 3 theme P. 1316:1317 arg1 strain 1328:1333 This AorCsxA-overexpressing P. pastoris strain 1288:1333 This AorCsxA-overexpressing P. pastoris strain 1288:1333 This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA. 26597529 6 4 theme acid 807:810 arg1 F769W 822:826 F769W 822:826 F769W 822:826 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 6 4 theme acid 807:810 arg1 mutation 812:819 A single amino acid mutation 792:819 A single amino acid mutation (F769W) in AorCsxA 792:838 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 3 5 theme exo-β-D-glucosaminidase 424:446 arg1 activity 455:462 exo-β-D-glucosaminidase (CsxA) activity 424:462 exo-β-D-glucosaminidase (CsxA) activity 424:462 AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. 26597529 9 6 theme large-scale 1351:1361 arg1 production 1363:1372 large-scale production 1351:1372 large-scale production of AorCsxA 1351:1383 This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA. 26597529 4 7 theme optimal 483:489 arg1 activity 491:498 optimal activity 483:498 optimal activity 483:498 AorCsxA exhibited optimal activity at pH 5.5 and 50°C; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50°C than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. 26597529 7 8 dep suggested 1027:1035 arg1 enabled 1086:1092 enabled 1086:1092 suggested the tightened pocket at the active site of F769W enabled efficient substrate binding 1027:1120 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 8 9 theme high-cell-density 1256:1272 arg1 fermentation 1274:1285 the high-cell-density fermentation 1252:1285 the high-cell-density fermentation 1252:1285 The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. 26597529 5 10 theme tested 773:778 arg1 ions 786:789 the tested metal ions 769:789 the tested metal ions 769:789 AorCsxA activity was inhibited by EDTA and most of the tested metal ions. 26597529 6 11 theme wild-type 977:985 arg1 enzyme 987:992 the wild-type enzyme 973:992 the wild-type enzyme 973:992 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 9 12 theme AorCsxA 1377:1383 arg1 production 1363:1372 large-scale production 1351:1372 large-scale production of AorCsxA 1351:1383 This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA. 26597529 7 13 theme substrate 1104:1112 arg1 binding 1114:1120 efficient substrate binding 1094:1120 efficient substrate binding 1094:1120 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 1 14 from oryzae 166:171 arg1 exo-β-D-glucosaminidase 115:137 An exo-β-D-glucosaminidase 112:137 An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 112:177 An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. 26597529 1 14 from oryzae 166:171 arg1 AorCsxA 140:146 AorCsxA 140:146 AorCsxA 140:146 An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. 26597529 5 15 theme AorCsxA 718:724 arg1 activity 726:733 AorCsxA activity 718:733 AorCsxA activity 718:733 AorCsxA activity was inhibited by EDTA and most of the tested metal ions. 26597529 0 16 theme Heterologous 0:11 arg1 Expression 13:22 Expression 13:22 Expression 13:22 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 8 17 theme AorCsxA 1127:1133 arg1 gene 1135:1138 The AorCsxA gene 1123:1138 The AorCsxA gene 1123:1138 The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. 26597529 6 18 theme amino 801:805 arg1 F769W 822:826 F769W 822:826 F769W 822:826 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 6 18 theme amino 801:805 arg1 mutation 812:819 A single amino acid mutation 792:819 A single amino acid mutation (F769W) in AorCsxA 792:838 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 4 19 theme stronger 593:600 arg1 thermostability 602:616 much stronger thermostability 588:616 much stronger thermostability 588:616 AorCsxA exhibited optimal activity at pH 5.5 and 50°C; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50°C than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. 26597529 6 20 theme single 794:799 arg1 F769W 822:826 F769W 822:826 F769W 822:826 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 6 20 theme single 794:799 arg1 mutation 812:819 A single amino acid mutation 792:819 A single amino acid mutation (F769W) in AorCsxA 792:838 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 3 21 theme CsxA 449:452 arg1 activity 455:462 exo-β-D-glucosaminidase (CsxA) activity 424:462 exo-β-D-glucosaminidase (CsxA) activity 424:462 AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. 26597529 7 22 theme efficient 1094:1102 arg1 binding 1114:1120 efficient substrate binding 1094:1120 efficient substrate binding 1094:1120 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 6 23 theme hydrolysis 889:898 arg1 velocity 900:907 hydrolysis velocity 889:907 hydrolysis velocity 889:907 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 8 24 theme U/ml 1231:1234 arg1 activity 1236:1243 222 U/ml activity 1227:1243 222 U/ml activity 1227:1243 The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. 26597529 0 25 from oryzae 104:109 arg1 Characterization 28:43 Characterization 28:43 Characterization 28:43 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 0 25 from oryzae 104:109 arg1 Expression 13:22 Expression 13:22 Expression 13:22 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 0 25 from oryzae 104:109 arg1 Exo-β-D-Glucosaminidase 63:85 a Thermostable Exo-β-D-Glucosaminidase 48:85 a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae 48:109 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 6 26 from mutation 812:819 arg1 AorCsxA 832:838 AorCsxA 832:838 AorCsxA 832:838 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 5 27 theme metal 780:784 arg1 ions 786:789 the tested metal ions 769:789 the tested metal ions 769:789 AorCsxA activity was inhibited by EDTA and most of the tested metal ions. 26597529 7 28 theme F769W 1080:1084 arg1 F769W 1080:1084 F769W 1080:1084 F769W 1080:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 7 28 theme F769W 1080:1084 arg1 site 1072:1075 the active site 1061:1075 the active site of F769W 1061:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 7 29 theme tightened 1041:1049 arg1 pocket 1051:1056 the tightened pocket 1037:1056 the tightened pocket at the active site of F769W 1037:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 0 30 theme Exo-β-D-Glucosaminidase 63:85 arg1 Characterization 28:43 Characterization 28:43 Characterization 28:43 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 0 30 theme Exo-β-D-Glucosaminidase 63:85 arg1 Expression 13:22 Expression 13:22 Expression 13:22 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 7 31 theme active 1065:1070 arg1 F769W 1080:1084 F769W 1080:1084 F769W 1080:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 7 31 theme active 1065:1070 arg1 site 1072:1075 the active site 1061:1075 the active site of F769W 1061:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 6 32 theme specific 867:874 arg1 activity 876:883 the specific activity 863:883 the specific activity 863:883 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 0 33 theme Thermostable 50:61 arg1 Exo-β-D-Glucosaminidase 63:85 a Thermostable Exo-β-D-Glucosaminidase 48:85 a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae 48:109 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 6 34 theme values 952:957 arg1 comparison 924:933 comparison 924:933 comparison of Vmax and kcat values with those of the wild-type enzyme 924:992 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 4 35 theme Pichia 553:558 arg1 PpAorCsxA 570:578 PpAorCsxA 570:578 PpAorCsxA 570:578 AorCsxA exhibited optimal activity at pH 5.5 and 50°C; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50°C than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. 26597529 4 35 theme Pichia 553:558 arg1 pastoris 560:567 Pichia pastoris 553:567 Pichia pastoris (PpAorCsxA) 553:579 AorCsxA exhibited optimal activity at pH 5.5 and 50°C; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50°C than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. 26597529 9 36 theme AorCsxA-overexpressing 1293:1314 arg1 strain 1328:1333 This AorCsxA-overexpressing P. pastoris strain 1288:1333 This AorCsxA-overexpressing P. pastoris strain 1288:1333 This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA. 26597529 6 37 with comparison 924:933 arg1 those 964:968 those 964:968 those 964:968 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 6 38 theme kcat 947:950 arg1 values 952:957 Vmax and kcat values 938:957 values 952:957 A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. 26597529 3 39 theme colloid 336:342 arg1 chitosan 344:351 colloid chitosan 336:351 colloid chitosan 336:351 AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. 26597529 2 40 theme acid 240:243 arg1 sequence 245:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. 26597529 1 41 theme Aspergillus 154:164 arg1 oryzae 166:171 Aspergillus oryzae FL402 154:177 Aspergillus oryzae FL402 154:177 An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. 26597529 2 42 theme amino 234:238 arg1 sequence 245:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. 26597529 2 43 theme hydrolase 299:307 arg1 family 309:314 glycoside hydrolase family 2 289:316 glycoside hydrolase family 2 289:316 The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. 26597529 7 44 theme three-dimensional 999:1015 arg1 structure 1017:1025 The three-dimensional structure 995:1025 The three-dimensional structure 995:1025 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 26597529 8 45 theme 222 1227:1229 arg1 U/ml 1231:1234 U/ml 1231:1234 U/ml 1231:1234 The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. 26597529 0 46 theme Aspergillus 92:102 arg1 oryzae 104:109 Aspergillus oryzae 92:109 Aspergillus oryzae 92:109 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 2 47 theme deduced 226:232 arg1 sequence 245:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence 222:252 The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. 26597529 2 48 theme glycoside 289:297 arg1 family 309:314 glycoside hydrolase family 2 289:316 glycoside hydrolase family 2 289:316 The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. 26597529 3 49 theme chitosan 383:390 arg1 oligosaccharides 392:407 chitosan oligosaccharides 383:407 chitosan oligosaccharides 383:407 AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. 26597529 0 50 from Characterization 28:43 arg1 oryzae 104:109 Aspergillus oryzae 92:109 Aspergillus oryzae 92:109 Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae. 26597529 7 51 from site 1072:1075 arg1 pocket 1051:1056 the tightened pocket 1037:1056 the tightened pocket at the active site of F769W 1037:1084 The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. 27351377 1 0 link monocyte-derived 269:284 arg1 macrophages 286:296 native monocyte-derived macrophages 262:296 native monocyte-derived macrophages 262:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 2 1 theme N- 348:349 arg1 alteration 334:343 the alteration 330:343 the alteration of N- and O-linked glycans as well as glycosphingolipids 330:400 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 1 2 theme immune 233:238 arg1 properties 240:249 several immune properties 225:249 several immune properties similar to native monocyte-derived macrophages 225:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 0 3 theme Line 79:82 arg1 Differentiation 39:53 the Differentiation 35:53 the Differentiation of Monocytic THP-1 Cell Line into Macrophages 35:99 Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages. 27351377 3 4 theme triantennary 695:706 arg1 structures 670:679 complex-type structures 657:679 complex-type structures 657:679 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 4 theme triantennary 695:706 arg1 glycans 708:714 triantennary glycans 695:714 particularly triantennary glycans 682:714 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 4 5 theme slight 793:798 arg1 decrease 800:807 a slight decrease 791:807 a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation 791:876 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 7 6 theme macrophagic 1507:1517 arg1 THP-1 1519:1523 monocytic and macrophagic THP-1 1493:1523 THP-1 1519:1523 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 3 7 theme Mass 510:513 arg1 spectrometry 515:526 Mass spectrometry 510:526 Mass spectrometry 510:526 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 8 theme macrophage 542:551 arg1 differentiation 553:567 macrophage differentiation 542:567 macrophage differentiation 542:567 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 5 9 from changes 893:899 arg1 glycosylation 904:916 glycosylation 904:916 glycosylation 904:916 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 1 10 theme similar 251:257 arg1 properties 240:249 several immune properties 225:249 several immune properties similar to native monocyte-derived macrophages 225:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 6 11 theme cell 1269:1272 arg1 surface 1274:1280 the cell surface 1265:1280 the cell surface of THP-1 macrophage-like cells 1265:1311 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 6 12 theme THP-1 1285:1289 arg1 cells 1307:1311 THP-1 macrophage-like cells 1285:1311 THP-1 macrophage-like cells 1285:1311 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 5 13 theme relevant 967:974 arg1 glycosyltransferases 976:995 relevant glycosyltransferases 967:995 relevant glycosyltransferases 967:995 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 3 14 from increase 645:652 arg1 structures 670:679 complex-type structures 657:679 complex-type structures 657:679 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 14 from increase 645:652 arg1 glycans 708:714 triantennary glycans 695:714 particularly triantennary glycans 682:714 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 5 15 theme gene 948:951 arg1 expression 953:962 gene expression 948:962 gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase 948:1117 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 4 16 theme α2,6-sialylation 861:876 arg1 proportion 816:825 the proportion 812:825 the proportion of multifucosylated N-glycans and α2,6-sialylation 812:876 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 5 17 theme uncovered 883:891 arg1 changes 893:899 The uncovered changes 879:899 The uncovered changes in glycosylation 879:916 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 2 18 link O-linked 355:362 arg1 glycans 364:370 O-linked glycans 355:370 O-linked glycans 355:370 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 6 19 theme glycan 1186:1191 arg1 structures 1193:1202 glycan structures 1186:1202 glycan structures 1186:1202 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 2 20 theme mass 443:446 arg1 spectrometry 448:459 mass spectrometry 443:459 mass spectrometry 443:459 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 2 21 theme real-time 495:503 arg1 PCR 505:507 quantitative real-time PCR 482:507 quantitative real-time PCR 482:507 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 5 22 theme glycosyltransferases 976:995 arg1 expression 953:962 gene expression 948:962 gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase 948:1117 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 0 23 theme Glycosylation 0:12 arg1 Changes 14:20 Glycosylation Changes 0:20 Glycosylation Changes 0:20 Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages. 27351377 1 24 theme native 262:267 arg1 macrophages 286:296 native monocyte-derived macrophages 262:296 native monocyte-derived macrophages 262:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 3 25 theme marked 578:583 arg1 upregulation 585:596 a marked upregulation 576:596 a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans 576:766 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 26 from upregulation 585:596 arg1 structures 670:679 complex-type structures 657:679 complex-type structures 657:679 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 26 from upregulation 585:596 arg1 glycans 708:714 triantennary glycans 695:714 particularly triantennary glycans 682:714 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 1 27 theme in 174:175 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 27 theme in 174:175 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 4 28 gly multifucosylated 830:845 arg1 N-glycans 847:855 multifucosylated N-glycans 830:855 multifucosylated N-glycans 830:855 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 1 29 theme monocyte-derived 269:284 arg1 macrophages 286:296 native monocyte-derived macrophages 262:296 native monocyte-derived macrophages 262:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 7 30 theme THP-1 1372:1376 arg1 maturation 1358:1367 macrophagic maturation 1346:1367 macrophagic maturation of THP-1 1346:1376 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 5 31 theme glycosidases 1001:1012 arg1 expression 953:962 gene expression 948:962 gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase 948:1117 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 2 32 theme THP-1 410:414 arg1 differentiation 416:430 THP-1 differentiation 410:430 THP-1 differentiation 410:430 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 5 33 theme expression 953:962 arg1 variations 934:943 variations 934:943 variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase 934:1117 The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. 27351377 3 34 theme high-mannose 745:756 arg1 N-glycans 758:766 high-mannose N-glycans 745:766 high-mannose N-glycans 745:766 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 2 35 theme O-linked 355:362 arg1 glycans 364:370 O-linked glycans 355:370 O-linked glycans 355:370 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 6 36 theme cells 1307:1311 arg1 surface 1274:1280 the cell surface 1265:1280 the cell surface of THP-1 macrophage-like cells 1265:1311 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 1 37 theme human 106:110 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 37 theme human 106:110 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 38 theme phagocytic 183:192 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 38 theme phagocytic 183:192 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 7 39 theme glycosylation 1424:1436 arg1 pattern 1438:1444 the surface glycosylation pattern 1412:1444 the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1412:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 3 40 theme N-glycans 758:766 arg1 expense 734:740 the expense 730:740 the expense of high-mannose N-glycans 730:766 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 7 41 theme surface 1416:1422 arg1 pattern 1438:1444 the surface glycosylation pattern 1412:1444 the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1412:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 3 42 theme expression 601:610 arg1 increase 645:652 an increase 642:652 a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans 576:766 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 42 theme expression 601:610 arg1 upregulation 585:596 a marked upregulation 576:596 a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans 576:766 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 2 43 theme glycans 364:370 arg1 alteration 334:343 the alteration 330:343 the alteration of N- and O-linked glycans as well as glycosphingolipids 330:400 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 1 44 theme cell 194:197 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 44 theme cell 194:197 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 2 45 theme flow 462:465 arg1 cytometry 467:475 flow cytometry 462:475 flow cytometry 462:475 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 1 46 dep in 174:175 arg1 vitro 177:181 vitro 177:181 vitro 177:181 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 7 47 theme THP-1 1519:1523 arg1 interaction 1478:1488 differential interaction 1465:1488 differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1465:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 2 48 theme glycosphingolipids 383:400 arg1 alteration 334:343 the alteration 330:343 the alteration of N- and O-linked glycans as well as glycosphingolipids 330:400 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 0 49 theme Monocytic 58:66 arg1 Line 79:82 Monocytic THP-1 Cell Line 58:82 Monocytic THP-1 Cell Line 58:82 Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages. 27351377 2 50 theme quantitative 482:493 arg1 PCR 505:507 quantitative real-time PCR 482:507 quantitative real-time PCR 482:507 In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. 27351377 4 51 theme N-glycans 847:855 arg1 proportion 816:825 the proportion 812:825 the proportion of multifucosylated N-glycans and α2,6-sialylation 812:876 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 1 52 theme acute 112:116 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 52 theme acute 112:116 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 4 53 theme multifucosylated 830:845 arg1 N-glycans 847:855 multifucosylated N-glycans 830:855 multifucosylated N-glycans 830:855 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 7 54 theme differential 1465:1476 arg1 interaction 1478:1488 differential interaction 1465:1488 differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1465:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 1 55 theme monocytic 118:126 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 55 theme monocytic 118:126 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 3 56 theme ganglioside 619:629 arg1 expression 601:610 expression 601:610 expression of GM3 ganglioside 601:629 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 7 57 theme monocytic 1493:1501 arg1 THP-1 1519:1523 monocytic and macrophagic THP-1 1493:1523 THP-1 1519:1523 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 7 58 theme immune 1530:1535 arg1 lectins 1550:1556 immune or bacterial lectins 1530:1556 immune or bacterial lectins 1530:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 1 59 theme leukemia 128:135 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 59 theme leukemia 128:135 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 6 60 theme macrophage-like 1291:1305 arg1 cells 1307:1311 THP-1 macrophage-like cells 1285:1311 THP-1 macrophage-like cells 1285:1311 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 7 61 with interaction 1478:1488 arg1 lectins 1550:1556 immune or bacterial lectins 1530:1556 immune or bacterial lectins 1530:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 3 62 theme complex-type 657:668 arg1 structures 670:679 complex-type structures 657:679 complex-type structures 657:679 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 3 62 theme complex-type 657:668 arg1 glycans 708:714 triantennary glycans 695:714 particularly triantennary glycans 682:714 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 1 63 dep model 199:203 arg1 exhibits 216:223 exhibits 216:223 exhibits several immune properties similar to native monocyte-derived macrophages 216:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 64 theme cell 137:140 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 64 theme cell 137:140 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 6 65 theme glycosylation 1241:1253 arg1 alteration 1227:1236 the alteration 1223:1236 the alteration of glycosylation 1223:1253 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 7 66 theme dramatic 1386:1393 arg1 modifications 1395:1407 dramatic modifications 1386:1407 dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1386:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 0 67 theme Cell 74:77 arg1 Line 79:82 Monocytic THP-1 Cell Line 58:82 Monocytic THP-1 Cell Line 58:82 Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages. 27351377 6 68 theme flow 1139:1142 arg1 cytometry 1144:1152 flow cytometry 1139:1152 flow cytometry 1139:1152 Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. 27351377 3 69 theme GM3 615:617 arg1 ganglioside 619:629 GM3 ganglioside 615:629 GM3 ganglioside 615:629 Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. 27351377 7 70 theme pattern 1438:1444 arg1 modifications 1395:1407 dramatic modifications 1386:1407 dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins 1386:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 1 71 theme line 142:145 arg1 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 71 theme line 142:145 arg1 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 7 72 theme macrophagic 1346:1356 arg1 maturation 1358:1367 macrophagic maturation 1346:1367 macrophagic maturation of THP-1 1346:1376 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 0 73 theme THP-1 68:72 arg1 Line 79:82 Monocytic THP-1 Cell Line 58:82 Monocytic THP-1 Cell Line 58:82 Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages. 27351377 1 74 used used 163:166 arg2 model 199:203 an in vitro phagocytic cell model 171:203 an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages 171:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 1 74 used used 163:166 arg2 THP-1 147:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 102:151 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 27351377 4 75 from decrease 800:807 arg1 proportion 816:825 the proportion 812:825 the proportion of multifucosylated N-glycans and α2,6-sialylation 812:876 Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. 27351377 7 76 theme bacterial 1540:1548 arg1 lectins 1550:1556 immune or bacterial lectins 1530:1556 immune or bacterial lectins 1530:1556 Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins. 27351377 1 77 theme several 225:231 arg1 properties 240:249 several immune properties 225:249 several immune properties similar to native monocyte-derived macrophages 225:296 The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. 26355101 1 0 theme several 356:362 arg1 diseases 364:371 several diseases 356:371 several diseases 356:371 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 0 1 theme Enterocytes 191:201 arg1 Glycome 163:169 the Cell Surface Glycome 146:169 the Cell Surface Glycome of Undifferentiated Enterocytes 146:201 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 3 2 theme present 550:556 arg1 Glycans 542:548 Glycans 542:548 Glycans present on the surface of intestinal epithelial cells (IEC) 542:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 6 3 theme border 1028:1033 arg1 membrane 1035:1042 a brush border membrane 1020:1042 a brush border membrane 1020:1042 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 2 4 theme cellular 516:523 arg1 transformations 525:539 cellular transformations 516:539 cellular transformations 516:539 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 0 5 from Changes 15:21 arg1 Glycosylation 39:51 Cell Surface Glycosylation 26:51 Cell Surface Glycosylation 26:51 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 0 6 theme Undifferentiated 174:189 arg1 Enterocytes 191:201 Undifferentiated Enterocytes 174:201 Undifferentiated Enterocytes 174:201 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 4 7 theme IECs 740:743 arg1 feature 729:735 A distinct feature 718:735 A distinct feature of IECs 718:743 A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. 26355101 6 8 theme brush 1022:1026 arg1 membrane 1035:1042 a brush border membrane 1020:1042 a brush border membrane 1020:1042 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 3 9 with interactions 618:629 arg1 billions 636:643 billions 636:643 billions 636:643 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 9 10 theme GI 1664:1665 arg1 tract 1667:1671 the GI tract 1660:1671 the GI tract 1660:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 6 11 from decrease 1047:1054 arg1 fucosylated 1114:1124 fucosylated 1114:1124 fucosylated 1114:1124 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 6 11 from decrease 1047:1054 arg1 glycans 1077:1083 high mannose type glycans 1059:1083 high mannose type glycans 1059:1083 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 6 11 from decrease 1047:1054 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 3 12 theme epithelial 587:596 arg1 IEC 605:607 IEC 605:607 IEC 605:607 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 3 12 theme epithelial 587:596 arg1 cells 598:602 intestinal epithelial cells 576:602 intestinal epithelial cells (IEC) 576:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 6 13 theme high 1059:1062 arg1 glycans 1077:1083 high mannose type glycans 1059:1083 high mannose type glycans 1059:1083 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 9 14 theme IEC 1559:1561 arg1 glycans 1571:1577 IEC surface glycans 1559:1577 IEC surface glycans 1559:1577 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 3 15 theme cells 598:602 arg1 surface 565:571 the surface 561:571 the surface of intestinal epithelial cells (IEC) 561:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 1 16 theme cells 294:298 arg1 differentiation 275:289 differentiation 275:289 differentiation 275:289 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 1 16 theme cells 294:298 arg1 development 259:269 development 259:269 development 259:269 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 0 17 theme High 113:116 arg1 Structures 126:135 High Mannose Structures 113:135 High Mannose Structures 113:135 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 7 18 theme cell 1264:1267 arg1 glycome 1277:1283 the cell surface glycome 1260:1283 the cell surface glycome 1260:1283 At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. 26355101 5 19 theme QTOF 862:865 arg1 profiling 870:878 nano-LC/ESI QTOF MS profiling 850:878 nano-LC/ESI QTOF MS profiling 850:878 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 2 20 theme extracellular 442:454 arg1 conditions 456:465 intra- and extracellular conditions 431:465 intra- and extracellular conditions 431:465 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 8 21 theme functional 1372:1381 arg1 role 1383:1386 a key functional role 1366:1386 a key functional role 1366:1386 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 6 22 theme mannose 1064:1070 arg1 glycans 1077:1083 high mannose type glycans 1059:1083 high mannose type glycans 1059:1083 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 9 23 theme Developing 1503:1512 arg1 methodologies 1514:1526 Developing methodologies 1503:1526 Developing methodologies to rapidly identify changes in IEC surface glycans 1503:1577 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 8 24 theme key 1368:1370 arg1 role 1383:1386 a key functional role 1366:1386 a key functional role 1366:1386 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 9 25 theme surface 1563:1569 arg1 glycans 1571:1577 IEC surface glycans 1559:1577 IEC surface glycans 1559:1577 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 0 26 theme Characteristic 0:13 arg1 Changes 15:21 Characteristic Changes 0:21 Characteristic Changes in Cell Surface Glycosylation 0:51 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 2 27 theme sensitivity 416:426 arg1 tool 497:500 an indispensable tool 480:500 an indispensable tool for analyzing cellular transformations 480:539 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 2 27 theme sensitivity 416:426 arg1 glycans 468:474 glycans 468:474 glycans 468:474 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 3 28 theme mammalian 703:711 arg1 gut 713:715 the mammalian gut 699:715 the mammalian gut 699:715 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 6 29 gly sialylated 1130:1139 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 9 30 theme screening 1599:1607 arg1 approach 1609:1616 a rapid screening approach 1591:1616 a rapid screening approach for a variety of disease states affecting the GI tract 1591:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 5 31 theme cell 960:963 arg1 differentiation 965:979 Caco-2 cell differentiation 953:979 Caco-2 cell differentiation 953:979 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 4 32 theme distinct 720:727 arg1 feature 729:735 A distinct feature 718:735 A distinct feature of IECs 718:743 A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. 26355101 1 33 theme diseases 364:371 arg1 progression 341:351 the progression 337:351 the progression of several diseases 337:371 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 6 34 theme type 1072:1075 arg1 glycans 1077:1083 high mannose type glycans 1059:1083 high mannose type glycans 1059:1083 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 2 35 theme diversity 402:410 arg1 tool 497:500 an indispensable tool 480:500 an indispensable tool for analyzing cellular transformations 480:539 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 2 35 theme diversity 402:410 arg1 glycans 468:474 glycans 468:474 glycans 468:474 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 0 36 theme Cell 26:29 arg1 Glycosylation 39:51 Cell Surface Glycosylation 26:51 Cell Surface Glycosylation 26:51 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 5 37 theme MS 867:868 arg1 profiling 870:878 nano-LC/ESI QTOF MS profiling 850:878 nano-LC/ESI QTOF MS profiling 850:878 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 8 38 theme glycans 1320:1326 arg1 expression 1306:1315 Differential expression 1293:1315 Differential expression of glycans during IEC maturation 1293:1348 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 5 39 from changes 909:915 arg1 glycosylation 920:932 glycosylation 920:932 glycosylation 920:932 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 5 40 used used 884:887 arg2 profiling 870:878 nano-LC/ESI QTOF MS profiling 850:878 nano-LC/ESI QTOF MS profiling 850:878 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 5 41 theme nano-LC/ESI 850:860 arg1 profiling 870:878 nano-LC/ESI QTOF MS profiling 850:878 nano-LC/ESI QTOF MS profiling 850:878 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 6 42 theme sialylated 1130:1139 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 8 43 theme membrane-associated 1406:1424 arg1 hydrolases 1426:1435 the membrane-associated hydrolases 1402:1435 the membrane-associated hydrolases 1402:1435 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 0 44 theme Mannose 118:124 arg1 Structures 126:135 High Mannose Structures 113:135 High Mannose Structures 113:135 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 8 45 theme defense 1483:1489 arg1 mechanisms 1491:1500 innate defense mechanisms 1476:1500 innate defense mechanisms 1476:1500 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 1 46 theme cell 215:218 arg1 glycosylation 228:240 cell surface glycosylation 215:240 cell surface glycosylation 215:240 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 6 47 theme Caco-2 985:990 arg1 cells 992:996 Caco-2 cells 985:996 Caco-2 cells 985:996 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 1 48 dep development 259:269 arg1 the 255:257 the 255:257 the 255:257 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 9 49 theme disease 1635:1641 arg1 states 1643:1648 disease states 1635:1648 disease states affecting the GI tract 1635:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 1 50 theme surface 220:226 arg1 glycosylation 228:240 cell surface glycosylation 215:240 cell surface glycosylation 215:240 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 0 51 theme Surface 31:37 arg1 Glycosylation 39:51 Cell Surface Glycosylation 26:51 Cell Surface Glycosylation 26:51 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 9 52 dep methodologies 1514:1526 arg1 identify 1539:1546 identify 1539:1546 to rapidly identify changes in IEC surface glycans 1528:1577 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 8 53 theme Differential 1293:1304 arg1 expression 1306:1315 Differential expression 1293:1315 Differential expression of glycans during IEC maturation 1293:1348 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 6 54 theme concurrent 1091:1100 arg1 increase 1102:1109 a concurrent increase 1089:1109 a concurrent increase in fucosylated and sialylated complex/hybrid type glycans 1089:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 9 55 from changes 1548:1554 arg1 glycans 1571:1577 IEC surface glycans 1559:1577 IEC surface glycans 1559:1577 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 2 56 theme intra- 431:436 arg1 conditions 456:465 intra- and extracellular conditions 431:465 intra- and extracellular conditions 431:465 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 0 57 theme Intestinal 63:72 arg1 IEC 91:93 IEC 91:93 IEC 91:93 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 0 57 theme Intestinal 63:72 arg1 Cell 85:88 Intestinal Epithelial Cell 63:88 Intestinal Epithelial Cell (IEC) Differentiation 63:110 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 0 58 theme Surface 155:161 arg1 Glycome 163:169 the Cell Surface Glycome 146:169 the Cell Surface Glycome of Undifferentiated Enterocytes 146:201 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 9 59 theme states 1643:1648 arg1 states 1643:1648 disease states 1635:1648 disease states affecting the GI tract 1635:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 9 59 theme states 1643:1648 arg1 variety 1624:1630 a variety 1622:1630 a variety of disease states affecting the GI tract 1622:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 8 60 theme IEC 1335:1337 arg1 maturation 1339:1348 IEC maturation 1335:1348 IEC maturation 1335:1348 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 2 61 theme indispensable 483:495 arg1 tool 497:500 an indispensable tool 480:500 an indispensable tool for analyzing cellular transformations 480:539 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 2 61 theme indispensable 483:495 arg1 glycans 468:474 glycans 468:474 glycans 468:474 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 0 62 theme Cell 150:153 arg1 Glycome 163:169 the Cell Surface Glycome 146:169 the Cell Surface Glycome of Undifferentiated Enterocytes 146:201 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 8 63 theme mucosal 1460:1466 arg1 surface 1468:1474 the mucosal surface 1456:1474 the mucosal surface 1456:1474 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 6 64 theme type 1156:1159 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 8 65 theme innate 1476:1481 arg1 mechanisms 1491:1500 innate defense mechanisms 1476:1500 innate defense mechanisms 1476:1500 Differential expression of glycans during IEC maturation appears to play a key functional role in regulating the membrane-associated hydrolases and contributes to the mucosal surface innate defense mechanisms. 26355101 7 66 theme surface 1269:1275 arg1 glycome 1277:1283 the cell surface glycome 1260:1283 the cell surface glycome 1260:1283 At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. 26355101 4 67 theme villus 823:828 arg1 tip 830:832 the villus tip 819:832 the villus tip 819:832 A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. 26355101 3 68 attach present 550:556 arg2 Glycans 542:548 Glycans 542:548 Glycans present on the surface of intestinal epithelial cells (IEC) 542:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 3 68 attach present 550:556 arg1 surface 565:571 the surface 561:571 the surface of intestinal epithelial cells (IEC) 561:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 6 69 theme complex/hybrid 1141:1154 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 3 70 theme intestinal 576:585 arg1 IEC 605:607 IEC 605:607 IEC 605:607 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 3 70 theme intestinal 576:585 arg1 cells 598:602 intestinal epithelial cells 576:602 intestinal epithelial cells (IEC) 576:608 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 4 71 theme crypt 805:809 arg1 base 811:814 the crypt base 801:814 the crypt base 801:814 A distinct feature of IECs is that they differentiate as they migrate upwards from the crypt base to the villus tip. 26355101 5 72 theme Caco-2 953:958 arg1 differentiation 965:979 Caco-2 cell differentiation 953:979 Caco-2 cell differentiation 953:979 In this study, nano-LC/ESI QTOF MS profiling was used to characterize the changes in glycosylation that correspond to Caco-2 cell differentiation. 26355101 7 73 theme glycome 1277:1283 arg1 remodeling 1246:1255 remodeling 1246:1255 remodeling of the cell surface glycome 1246:1283 At day 21, when cells appear to be completely differentiated, remodeling of the cell surface glycome ceases. 26355101 0 74 theme Cell 85:88 arg1 Differentiation 96:110 Intestinal Epithelial Cell (IEC) Differentiation 63:110 Intestinal Epithelial Cell (IEC) Differentiation 63:110 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 3 75 theme native 648:653 arg1 microorganisms 655:668 native microorganisms 648:668 native microorganisms 648:668 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 1 76 from Changes 204:210 arg1 glycosylation 228:240 cell surface glycosylation 215:240 cell surface glycosylation 215:240 Changes in cell surface glycosylation occur during the development and differentiation of cells and have been widely correlated with the progression of several diseases. 26355101 2 77 theme structural 391:400 arg1 diversity 402:410 their structural diversity 385:410 their structural diversity 385:410 Because of their structural diversity and sensitivity to intra- and extracellular conditions, glycans are an indispensable tool for analyzing cellular transformations. 26355101 0 78 theme Epithelial 74:83 arg1 IEC 91:93 IEC 91:93 IEC 91:93 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 0 78 theme Epithelial 74:83 arg1 Cell 85:88 Intestinal Epithelial Cell 63:88 Intestinal Epithelial Cell (IEC) Differentiation 63:110 Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. 26355101 3 79 theme microorganisms 655:668 arg1 billions 636:643 billions 636:643 billions 636:643 Glycans present on the surface of intestinal epithelial cells (IEC) mediate interactions with billions of native microorganisms, which continuously populate the mammalian gut. 26355101 9 80 theme rapid 1593:1597 arg1 approach 1609:1616 a rapid screening approach 1591:1616 a rapid screening approach for a variety of disease states affecting the GI tract 1591:1671 Developing methodologies to rapidly identify changes in IEC surface glycans may lead to a rapid screening approach for a variety of disease states affecting the GI tract. 26355101 6 81 from increase 1102:1109 arg1 fucosylated 1114:1124 fucosylated 1114:1124 fucosylated 1114:1124 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 6 81 from increase 1102:1109 arg1 glycans 1077:1083 high mannose type glycans 1059:1083 high mannose type glycans 1059:1083 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 26355101 6 81 from increase 1102:1109 arg1 glycans 1161:1167 sialylated complex/hybrid type glycans 1130:1167 sialylated complex/hybrid type glycans 1130:1167 As Caco-2 cells differentiate to form a brush border membrane, a decrease in high mannose type glycans and a concurrent increase in fucosylated and sialylated complex/hybrid type glycans were observed. 27591901 8 0 theme Con 1036:1038 arg1 A 1040:1040 Con A 1036:1040 Con A 1036:1040 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 5 1 theme such 509:512 arg1 microscopy 545:554 the atomic force microscopy 528:554 the atomic force microscopy (AFM) working 528:568 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 1 theme such 509:512 arg1 method 514:519 One such method 505:519 One such method 505:519 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 8 2 theme cell-dependent 1180:1193 arg1 manner 1195:1200 a cell-dependent manner 1178:1200 a cell-dependent manner 1178:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 0 3 theme cells 55:59 arg1 distinction 31:41 the distinction 27:41 the distinction of melanoma cells with a different metastatic potential 27:97 AFM and QCM-D as tools for the distinction of melanoma cells with a different metastatic potential. 27591901 4 4 theme tumour 485:490 arg1 progression 492:502 tumour progression 485:502 tumour progression 485:502 In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. 27591901 8 5 attach presence 1130:1137 arg1 manner 1195:1200 a cell-dependent manner 1178:1200 a cell-dependent manner 1178:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 8 5 attach presence 1130:1137 arg2 groups 1168:1173 various glycan structural groups 1142:1173 various glycan structural groups 1142:1173 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 6 6 theme type 760:763 arg1 glycans 765:771 mannose type glycans 752:771 mannose type glycans 752:771 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 7 theme melanoma 802:809 arg1 cells 811:815 melanoma cells 802:815 melanoma cells 802:815 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 0 8 theme different 68:76 arg1 potential 89:97 a different metastatic potential 66:97 a different metastatic potential 66:97 AFM and QCM-D as tools for the distinction of melanoma cells with a different metastatic potential. 27591901 6 9 theme radial 838:843 arg1 WM35 859:862 WM35 859:862 WM35 859:862 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 9 theme radial 838:843 arg1 phase 852:856 the radial growth phase 834:856 the radial growth phase (WM35) 834:863 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 5 10 theme quartz 615:620 arg1 QCM 644:646 QCM 644:646 QCM 644:646 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 10 theme quartz 615:620 arg1 microbalance 630:641 quartz crystal microbalance 615:641 quartz crystal microbalance (QCM) 615:647 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 6 11 theme mannose 752:758 arg1 glycans 765:771 mannose type glycans 752:771 mannose type glycans 752:771 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 9 12 theme glucose 1378:1384 arg1 types 1386:1390 glucose types 1378:1390 glucose types of oligosaccharides 1378:1410 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 8 13 theme surface 1047:1053 arg1 glycans 1055:1061 surface glycans 1047:1061 surface glycans 1047:1061 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 5 14 theme crystal 622:628 arg1 QCM 644:646 QCM 644:646 QCM 644:646 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 14 theme crystal 622:628 arg1 microbalance 630:641 quartz crystal microbalance 615:641 quartz crystal microbalance (QCM) 615:647 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 2 15 theme therapy 264:270 arg1 efficiency 272:281 the therapy efficiency 260:281 the therapy efficiency 260:281 Thus, an early and proper melanoma diagnosis influences significantly the therapy efficiency. 27591901 9 16 theme A375-P 1322:1327 arg1 cells 1329:1333 A375-P cells 1322:1333 A375-P cells 1322:1333 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 8 17 theme structural 1157:1166 arg1 groups 1168:1173 various glycan structural groups 1142:1173 various glycan structural groups 1142:1173 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 5 18 theme molecular 679:687 arg1 interactions 689:700 the molecular interactions 675:700 the molecular interactions 675:700 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 4 19 theme melanoma 441:448 arg1 diagnosis 450:458 melanoma diagnosis 441:458 melanoma diagnosis 441:458 In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. 27591901 8 20 theme AFM 1089:1091 arg1 techniques 1101:1110 both AFM and QCM techniques 1084:1110 both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner 1084:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 1 21 theme dangerous 138:146 arg1 cancer 153:158 the most dangerous skin cancer 129:158 the most dangerous skin cancer originating from melanocytes 129:187 Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. 27591901 0 22 theme metastatic 78:87 arg1 potential 89:97 a different metastatic potential 66:97 a different metastatic potential 66:97 AFM and QCM-D as tools for the distinction of melanoma cells with a different metastatic potential. 27591901 5 23 theme force 577:581 arg1 spectroscopy 583:594 force spectroscopy 577:594 the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions 573:700 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 24 theme atomic 532:537 arg1 microscopy 545:554 the atomic force microscopy 528:554 the atomic force microscopy (AFM) working 528:568 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 24 theme atomic 532:537 arg1 method 514:519 One such method 505:519 One such method 505:519 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 24 theme atomic 532:537 arg1 AFM 557:559 AFM 557:559 AFM 557:559 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 9 25 theme oligosaccharides 1395:1410 arg1 mannose 1366:1372 long, branched mannose 1351:1372 long, branched mannose 1351:1372 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 9 25 theme oligosaccharides 1395:1410 arg1 types 1386:1390 glucose types 1378:1390 glucose types of oligosaccharides 1378:1410 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 1 26 theme skin 148:151 arg1 cancer 153:158 the most dangerous skin cancer 129:158 the most dangerous skin cancer originating from melanocytes 129:187 Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. 27591901 8 27 theme A 1040:1040 arg1 interactions 1020:1031 The interactions 1016:1031 The interactions of Con A with surface glycans 1016:1061 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 5 28 theme spectroscopy 583:594 arg1 mode 596:599 the force spectroscopy mode 573:599 the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions 573:700 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 29 theme force 539:543 arg1 microscopy 545:554 the atomic force microscopy 528:554 the atomic force microscopy (AFM) working 528:568 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 29 theme force 539:543 arg1 method 514:519 One such method 505:519 One such method 505:519 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 5 29 theme force 539:543 arg1 AFM 557:559 AFM 557:559 AFM 557:559 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 9 30 theme Con 1207:1209 arg1 A 1211:1211 The Con A 1203:1211 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 1203:1411 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 1 31 theme cancer 153:158 arg1 cancer 153:158 the most dangerous skin cancer 129:158 the most dangerous skin cancer originating from melanocytes 129:187 Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. 27591901 1 31 theme cancer 153:158 arg1 one 122:124 one 122:124 one 122:124 Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. 27591901 6 32 theme growth 845:850 arg1 WM35 859:862 WM35 859:862 WM35 859:862 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 32 theme growth 845:850 arg1 phase 852:856 the radial growth phase 834:856 the radial growth phase (WM35) 834:863 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 5 33 dep combined 601:608 arg1 both 650:653 both 650:653 both 650:653 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 7 34 from level 918:922 arg1 surfaces 933:940 their surfaces 927:940 their surfaces 927:940 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 3 35 theme subjective 354:363 arg1 assessments 365:375 subjective assessments 354:375 subjective assessments 354:375 The melanoma recognition is still difficult, and generally, relies on subjective assessments. 27591901 7 36 theme Con 982:984 arg1 A 979:979 lectin concanavalin A 959:979 lectin concanavalin A (Con A) from Canavalia ensiformis 959:1013 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 7 36 theme Con 982:984 arg1 A 986:986 Con A 982:986 Con A 982:986 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 4 37 theme methods 425:431 arg1 lack 404:407 a lack 402:407 a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression 402:502 In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. 27591901 8 38 theme glycan 1150:1155 arg1 groups 1168:1173 various glycan structural groups 1142:1173 various glycan structural groups 1142:1173 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 6 39 theme lung 874:877 arg1 metastasis 879:888 lung metastasis 874:888 lung metastasis (A375-P) 874:897 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 39 theme lung 874:877 arg1 A375-P 891:896 A375-P 891:896 A375-P 891:896 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 7 40 theme concanavalin 966:977 arg1 A 979:979 lectin concanavalin A 959:979 lectin concanavalin A (Con A) from Canavalia ensiformis 959:1013 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 7 40 theme concanavalin 966:977 arg1 A 986:986 Con A 982:986 Con A 982:986 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 4 41 theme quantitative 412:423 arg1 methods 425:431 quantitative methods 412:431 quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression 412:502 In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. 27591901 5 42 dep mode 596:599 arg1 combined 601:608 combined 601:608 combined with quartz crystal microbalance (QCM) 601:647 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 7 43 from ensiformis 1004:1013 arg1 A 979:979 lectin concanavalin A 959:979 lectin concanavalin A (Con A) from Canavalia ensiformis 959:1013 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 7 43 from ensiformis 1004:1013 arg1 A 986:986 Con A 982:986 Con A 982:986 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 8 44 theme QCM 1097:1099 arg1 techniques 1101:1110 both AFM and QCM techniques 1084:1110 both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner 1084:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 9 45 theme Con 1336:1338 arg1 A 1340:1340 Con A 1336:1340 Con A 1336:1340 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 2 46 theme early 199:203 arg1 diagnosis 225:233 an early and proper melanoma diagnosis 196:233 an early and proper melanoma diagnosis 196:233 Thus, an early and proper melanoma diagnosis influences significantly the therapy efficiency. 27591901 8 47 theme groups 1168:1173 arg1 presence 1130:1137 the presence 1126:1137 the presence of various glycan structural groups in a cell-dependent manner 1126:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 4 48 theme progression 492:502 arg1 monitoring 471:480 the monitoring 467:480 the monitoring of tumour progression 467:502 In particular, there is a lack of quantitative methods used in melanoma diagnosis and in the monitoring of tumour progression. 27591901 3 49 theme melanoma 288:295 arg1 difficult 318:326 difficult 318:326 difficult 318:326 The melanoma recognition is still difficult, and generally, relies on subjective assessments. 27591901 3 49 theme melanoma 288:295 arg1 recognition 297:307 The melanoma recognition 284:307 The melanoma recognition 284:307 The melanoma recognition is still difficult, and generally, relies on subjective assessments. 27591901 9 50 dep long 1351:1354 arg1 branched 1357:1364 branched 1357:1364 branched 1357:1364 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 5 51 from microscopy 545:554 arg1 mode 596:599 the force spectroscopy mode 573:599 the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions 573:700 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 7 52 theme lectin 959:964 arg1 A 979:979 lectin concanavalin A 959:979 lectin concanavalin A (Con A) from Canavalia ensiformis 959:1013 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 7 52 theme lectin 959:964 arg1 A 986:986 Con A 982:986 Con A 982:986 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 7 53 theme Canavalia 994:1002 arg1 ensiformis 1004:1013 Canavalia ensiformis 994:1013 Canavalia ensiformis 994:1013 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 8 54 from presence 1130:1137 arg1 manner 1195:1200 a cell-dependent manner 1178:1200 a cell-dependent manner 1178:1200 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 1 55 theme Malignant 100:108 arg1 melanoma 110:117 Malignant melanoma 100:117 Malignant melanoma 100:117 Malignant melanoma is one of the most dangerous skin cancer originating from melanocytes. 27591901 9 56 theme present 1249:1255 arg1 mannose 1215:1221 mannose (or glucose) type glycans 1215:1247 mannose 1215:1221 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 9 57 theme glucose 1227:1233 arg1 glycans 1241:1247 mannose (or glucose) type glycans 1215:1247 glycans 1241:1247 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 8 58 theme various 1142:1148 arg1 groups 1168:1173 various glycan structural groups 1142:1173 various glycan structural groups 1142:1173 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 9 59 dep A 1211:1211 arg1 short 1289:1293 short 1289:1293 short 1289:1293 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 9 59 dep A 1211:1211 arg1 ramified 1304:1311 ramified 1304:1311 ramified 1304:1311 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 0 60 with distinction 31:41 arg1 potential 89:97 a different metastatic potential 66:97 a different metastatic potential 66:97 AFM and QCM-D as tools for the distinction of melanoma cells with a different metastatic potential. 27591901 9 61 theme long 1351:1354 arg1 mannose 1366:1372 long, branched mannose 1351:1372 long, branched mannose 1351:1372 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 0 62 theme melanoma 46:53 arg1 cells 55:59 melanoma cells 46:59 melanoma cells 46:59 AFM and QCM-D as tools for the distinction of melanoma cells with a different metastatic potential. 27591901 6 63 theme glycans 765:771 arg1 recognition 737:747 the recognition 733:747 the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P) 733:897 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 2 64 theme melanoma 216:223 arg1 diagnosis 225:233 an early and proper melanoma diagnosis 196:233 an early and proper melanoma diagnosis 196:233 Thus, an early and proper melanoma diagnosis influences significantly the therapy efficiency. 27591901 7 65 theme glycosylation 904:916 arg1 level 918:922 The glycosylation level 900:922 The glycosylation level on their surfaces 900:940 The glycosylation level on their surfaces was probed using lectin concanavalin A (Con A) from Canavalia ensiformis. 27591901 8 66 with interactions 1020:1031 arg1 glycans 1055:1061 surface glycans 1047:1061 surface glycans 1047:1061 The interactions of Con A with surface glycans were quantified with both AFM and QCM techniques that revealed the presence of various glycan structural groups in a cell-dependent manner. 27591901 6 67 from recognition 737:747 arg1 melanocytes 776:786 melanocytes 776:786 melanocytes (HEMa-LP) 776:796 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 67 from recognition 737:747 arg1 cells 811:815 melanoma cells 802:815 melanoma cells 802:815 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 6 67 from recognition 737:747 arg1 HEMa-LP 789:795 HEMa-LP 789:795 HEMa-LP 789:795 In our study we have compared the recognition of mannose type glycans in melanocytes (HEMa-LP) and melanoma cells originating from the radial growth phase (WM35) and from lung metastasis (A375-P). 27591901 9 68 theme cell 1265:1268 arg1 surface 1270:1276 WM35 cell surface 1260:1276 WM35 cell surface 1260:1276 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 2 69 theme proper 209:214 arg1 diagnosis 225:233 an early and proper melanoma diagnosis 196:233 an early and proper melanoma diagnosis 196:233 Thus, an early and proper melanoma diagnosis influences significantly the therapy efficiency. 27591901 9 70 attach present 1249:1255 arg2 mannose 1215:1221 mannose (or glucose) type glycans 1215:1247 mannose 1215:1221 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 9 70 attach present 1249:1255 arg1 surface 1270:1276 WM35 cell surface 1260:1276 WM35 cell surface 1260:1276 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 27591901 5 71 dep microscopy 545:554 arg1 working 562:568 working 562:568 working 562:568 One such method can be the atomic force microscopy (AFM) working in the force spectroscopy mode combined with quartz crystal microbalance (QCM), both applied to quantify the molecular interactions. 27591901 9 72 theme type 1236:1239 arg1 glycans 1241:1247 mannose (or glucose) type glycans 1215:1247 glycans 1241:1247 The Con A - mannose (or glucose) type glycans present on WM35 cell surface are rather short and less ramified while in A375-P cells, Con A binds to long, branched mannose and glucose types of oligosaccharides. 26509638 0 0 theme Arabinomannan 84:96 arg1 Fragments 98:106 Mycobacterial Arabinomannan Fragments 70:106 Mycobacterial Arabinomannan Fragments 70:106 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 3 1 theme mycobacterial 494:506 arg1 fragments 522:530 mycobacterial arabinomannan fragments 494:530 mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method 494:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 4 2 from core 685:688 arg1 present 670:676 present 670:676 present 670:676 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 0 3 theme Mycobacterial 70:82 arg1 Fragments 98:106 Mycobacterial Arabinomannan Fragments 70:106 Mycobacterial Arabinomannan Fragments 70:106 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 2 4 from patients 341:348 arg1 present 330:336 present 330:336 present 330:336 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 3 5 theme arabinomannan 508:520 arg1 fragments 522:530 mycobacterial arabinomannan fragments 494:530 mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method 494:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 2 6 from present 330:336 arg1 patients 341:348 patients 341:348 patients infected by mycobacteria 341:373 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 1 7 theme immunogenic 252:262 arg1 antigens 264:271 immunogenic antigens 252:271 immunogenic antigens 252:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 7 8 theme selective 1154:1162 arg1 removal 1164:1170 selective removal 1154:1170 selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue 1154:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 9 theme mannopyranose 1338:1350 arg1 trichloroacetimidate 1352:1371 a mannopyranose trichloroacetimidate 1336:1371 a mannopyranose trichloroacetimidate 1336:1371 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 1 10 theme antigens 264:271 arg1 number 242:247 a number 240:247 a number of immunogenic antigens 240:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 0 11 theme Fragments 98:106 arg1 Synthesis 57:65 the Synthesis 53:65 the Synthesis of Mycobacterial Arabinomannan Fragments 53:106 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 4 12 from present 670:676 arg1 core 685:688 the core 681:688 the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 681:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 7 13 theme protecting 1179:1188 arg1 group 1190:1194 the protecting group 1175:1194 the protecting group present at the O-2 position of a single mannopyranoside residue 1175:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 4 14 theme key 697:699 arg1 LAM 747:749 LAM 747:749 LAM 747:749 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 14 theme key 697:699 arg1 lipoarabinomannan 728:744 the key immunogenic polysaccharide lipoarabinomannan 693:744 the key immunogenic polysaccharide lipoarabinomannan (LAM) 693:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 15 theme present 670:676 arg1 motifs 663:668 structural motifs 652:668 structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 652:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 16 theme structural 652:661 arg1 motifs 663:668 structural motifs 652:668 structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 652:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 7 17 theme group 1190:1194 arg1 removal 1164:1170 selective removal 1154:1170 selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue 1154:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 2 18 theme Circulating 274:284 arg1 antibodies 286:295 Circulating antibodies 274:295 Circulating antibodies that recognize these glycans 274:324 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 7 19 theme O-2 1211:1213 arg1 position 1215:1222 the O-2 position 1207:1222 the O-2 position of a single mannopyranoside residue 1207:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 2 20 theme antibodies 395:404 arg1 basis 419:423 the basis 415:423 the basis for new TB diagnostics 415:446 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 2 20 theme antibodies 395:404 arg1 detection 376:384 detection 376:384 detection of these antibodies 376:404 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 5 21 theme efficient 796:804 arg1 strategy 828:835 an efficient orthogonal protection strategy 793:835 an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7) 793:901 To access these compounds, we developed an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7). 26509638 7 22 with glycosylation 1273:1285 arg1 thioglycoside 1315:1327 a pentaarabinofuranose thioglycoside 1292:1327 a pentaarabinofuranose thioglycoside 1292:1327 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 22 with glycosylation 1273:1285 arg1 trichloroacetimidate 1352:1371 a mannopyranose trichloroacetimidate 1336:1371 a mannopyranose trichloroacetimidate 1336:1371 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 4 23 theme lipoarabinomannan 728:744 arg1 core 685:688 the core 681:688 the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 681:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 0 24 theme Orthogonal 18:27 arg1 Strategy 40:47 an Orthogonal Protection Strategy 15:47 an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments 15:106 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 2 25 located present 330:336 arg1 patients 341:348 patients 341:348 patients infected by mycobacteria 341:373 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 2 25 located present 330:336 arg2 antibodies 286:295 Circulating antibodies 274:295 Circulating antibodies that recognize these glycans 274:324 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 7 26 theme mannopyranoside 1236:1250 arg1 residue 1252:1258 a single mannopyranoside residue 1227:1258 a single mannopyranoside residue 1227:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 1 27 theme Mycobacterium 109:121 arg1 organism 141:148 the organism 137:148 the organism that causes tuberculosis (TB) 137:178 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 1 27 theme Mycobacterium 109:121 arg1 tuberculosis 123:134 Mycobacterium tuberculosis 109:134 Mycobacterium tuberculosis 109:134 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 2 28 theme new 429:431 arg1 diagnostics 436:446 new TB diagnostics 429:446 new TB diagnostics 429:446 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 2 29 theme TB 433:434 arg1 diagnostics 436:446 new TB diagnostics 429:446 new TB diagnostics 429:446 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 7 30 theme single 1229:1234 arg1 residue 1252:1258 a single mannopyranoside residue 1227:1258 a single mannopyranoside residue 1227:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 1 31 theme cell 205:208 arg1 structure 215:223 a carbohydrate-rich cell wall structure 185:223 a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens 185:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 0 32 theme Strategy 40:47 arg1 Development 0:10 Development 0:10 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 0:107 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 1 33 contain has 181:183 arg2 structure 215:223 a carbohydrate-rich cell wall structure 185:223 a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens 185:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 1 33 contain has 181:183 arg1 organism 141:148 the organism 137:148 the organism that causes tuberculosis (TB) 137:178 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 1 33 contain has 181:183 arg1 tuberculosis 123:134 Mycobacterium tuberculosis 109:134 Mycobacterium tuberculosis 109:134 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 1 34 theme wall 210:213 arg1 structure 215:223 a carbohydrate-rich cell wall structure 185:223 a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens 185:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 0 35 theme Protection 29:38 arg1 Strategy 40:47 an Orthogonal Protection Strategy 15:47 an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments 15:106 Development of an Orthogonal Protection Strategy for the Synthesis of Mycobacterial Arabinomannan Fragments. 26509638 5 36 theme arabinomannan 866:878 arg1 fragments 880:888 seven arabinomannan fragments 860:888 seven arabinomannan fragments of LAM (1-7) 860:901 To access these compounds, we developed an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7). 26509638 3 37 theme diagnostic 604:613 arg1 method 615:620 such a diagnostic method 597:620 such a diagnostic method 597:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 5 38 theme orthogonal 806:815 arg1 strategy 828:835 an efficient orthogonal protection strategy 793:835 an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7) 793:901 To access these compounds, we developed an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7). 26509638 7 39 theme present 1196:1202 arg1 group 1190:1194 the protecting group 1175:1194 the protecting group present at the O-2 position of a single mannopyranoside residue 1175:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 40 theme protected 1046:1054 arg1 9 1092:1092 9 1092:1092 9 1092:1092 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 40 theme protected 1046:1054 arg1 8 1087:1087 8 1087:1087 8 1087:1087 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 40 theme protected 1046:1054 arg1 derivative 1075:1084 a differentially protected trimannopyranoside derivative 1029:1084 a differentially protected trimannopyranoside derivative (8 or 9) 1029:1093 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 2 41 attach present 330:336 arg1 patients 341:348 patients 341:348 patients infected by mycobacteria 341:373 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 2 41 attach present 330:336 arg2 antibodies 286:295 Circulating antibodies 274:295 Circulating antibodies that recognize these glycans 274:324 Circulating antibodies that recognize these glycans are present in patients infected by mycobacteria; detection of these antibodies could be the basis for new TB diagnostics. 26509638 1 42 contain possesses 230:238 arg2 number 242:247 a number 240:247 a number of immunogenic antigens 240:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 1 42 contain possesses 230:238 arg1 structure 215:223 a carbohydrate-rich cell wall structure 185:223 a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens 185:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 7 43 attach present 1196:1202 arg1 position 1215:1222 the O-2 position 1207:1222 the O-2 position of a single mannopyranoside residue 1207:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 43 attach present 1196:1202 arg2 group 1190:1194 the protecting group 1175:1194 the protecting group present at the O-2 position of a single mannopyranoside residue 1175:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 3 44 theme panel 485:489 arg1 synthesis 470:478 the synthesis 466:478 the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method 466:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 7 45 theme trimannopyranoside 1056:1073 arg1 9 1092:1092 9 1092:1092 9 1092:1092 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 45 theme trimannopyranoside 1056:1073 arg1 8 1087:1087 8 1087:1087 8 1087:1087 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 45 theme trimannopyranoside 1056:1073 arg1 derivative 1075:1084 a differentially protected trimannopyranoside derivative 1029:1084 a differentially protected trimannopyranoside derivative (8 or 9) 1029:1093 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 7 46 theme residue 1252:1258 arg1 position 1215:1222 the O-2 position 1207:1222 the O-2 position of a single mannopyranoside residue 1207:1258 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 4 47 theme polysaccharide 713:726 arg1 LAM 747:749 LAM 747:749 LAM 747:749 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 47 theme polysaccharide 713:726 arg1 lipoarabinomannan 728:744 the key immunogenic polysaccharide lipoarabinomannan 693:744 the key immunogenic polysaccharide lipoarabinomannan (LAM) 693:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 7 48 from position 1215:1222 arg1 present 1196:1202 present 1196:1202 present 1196:1202 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 4 49 theme immunogenic 701:711 arg1 LAM 747:749 LAM 747:749 LAM 747:749 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 49 theme immunogenic 701:711 arg1 lipoarabinomannan 728:744 the key immunogenic polysaccharide lipoarabinomannan 693:744 the key immunogenic polysaccharide lipoarabinomannan (LAM) 693:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 50 attach present 670:676 arg1 core 685:688 the core 681:688 the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 681:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 4 50 attach present 670:676 arg2 motifs 663:668 structural motifs 652:668 structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM) 652:750 In this study, we focused on structural motifs present in the core of the key immunogenic polysaccharide lipoarabinomannan (LAM). 26509638 3 51 theme method 615:620 arg1 feasibility 582:592 the feasibility 578:592 the feasibility of such a diagnostic method 578:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 1 52 theme carbohydrate-rich 187:203 arg1 structure 215:223 a carbohydrate-rich cell wall structure 185:223 a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens 185:271 Mycobacterium tuberculosis, the organism that causes tuberculosis (TB), has a carbohydrate-rich cell wall structure that possesses a number of immunogenic antigens. 26509638 3 53 theme fragments 522:530 arg1 panel 485:489 a panel 483:489 a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method 483:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 26509638 7 54 theme pentaarabinofuranose 1294:1313 arg1 thioglycoside 1315:1327 a pentaarabinofuranose thioglycoside 1292:1327 a pentaarabinofuranose thioglycoside 1292:1327 Starting from a differentially protected trimannopyranoside derivative (8 or 9), the targets were obtained using an approach that involved selective removal of the protecting group present at the O-2 position of a single mannopyranoside residue, followed by glycosylation with a pentaarabinofuranose thioglycoside and/or a mannopyranose trichloroacetimidate. 26509638 5 55 theme protection 817:826 arg1 strategy 828:835 an efficient orthogonal protection strategy 793:835 an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7) 793:901 To access these compounds, we developed an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7). 26509638 5 56 theme LAM 893:895 arg1 fragments 880:888 seven arabinomannan fragments 860:888 seven arabinomannan fragments of LAM (1-7) 860:901 To access these compounds, we developed an efficient orthogonal protection strategy that allowed access to seven arabinomannan fragments of LAM (1-7). 26509638 3 57 from use 536:538 arg1 investigations 543:556 investigations 543:556 investigations directed at testing the feasibility of such a diagnostic method 543:620 We describe here the synthesis of a panel of mycobacterial arabinomannan fragments for use in investigations directed at testing the feasibility of such a diagnostic method. 24726881 2 0 theme important 128:136 arg1 role 138:141 An important role 125:141 An important role in cancer pathogenesis 125:164 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 5 1 theme GnT-V 713:717 arg1 overexpression 684:697 induced overexpression 676:697 induced overexpression of GnT-III or GnT-V 676:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 7 2 dep 2-aminobenzamide 960:975 arg1 adapted 978:984 adapted 978:984 adapted from Taniguchi et al., 1989 978:1012 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 13 3 from mechanisms 1695:1704 arg1 progression 1778:1788 cancer progression 1771:1788 cancer progression 1771:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 11 4 theme bisected 1436:1443 arg1 N-glycans 1446:1454 "bisected" N-glycans 1435:1454 "bisected" N-glycans 1435:1454 To the best of our knowledge, the result of our study is the first to demonstrate that "bisected" N-glycans can be carried by MCAM. 24726881 9 5 located present 1156:1162 arg1 molecule 1190:1197 melanoma cell adhesion molecule 1167:1197 melanoma cell adhesion molecule (known as MCAM/MUC18) 1167:1219 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 5 located present 1156:1162 arg2 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 5 located present 1156:1162 arg2 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 5 located present 1156:1162 arg2 amounts 1098:1104 Increased amounts 1088:1104 Increased amounts of "bisected" and beta1-6 branched N-glycans 1088:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 5 6 theme GnT-III 702:708 arg1 overexpression 684:697 induced overexpression 676:697 induced overexpression of GnT-III or GnT-V 676:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 7 7 theme novel 898:902 arg1 method 909:914 a novel HPLC method 896:914 a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989) 896:1013 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 8 8 dep Higher 1016:1021 arg1 activities 1038:1047 activities 1038:1047 activities 1038:1047 Higher expression and activities of glycosyltransferases were detected. 24726881 8 8 dep Higher 1016:1021 arg1 expression 1023:1032 expression 1023:1032 expression 1023:1032 Higher expression and activities of glycosyltransferases were detected. 24726881 11 9 theme study 1396:1400 arg1 result 1382:1387 the result 1378:1387 the result of our study 1378:1400 To the best of our knowledge, the result of our study is the first to demonstrate that "bisected" N-glycans can be carried by MCAM. 24726881 11 9 theme study 1396:1400 arg1 first 1409:1413 first 1409:1413 first 1409:1413 To the best of our knowledge, the result of our study is the first to demonstrate that "bisected" N-glycans can be carried by MCAM. 24726881 5 10 theme study 618:622 arg1 aim 606:608 The aim 602:608 The aim of this study 602:622 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 14 11 theme based 1914:1918 arg1 systems 1925:1931 2-aminopyridine based HPLC systems 1898:1931 2-aminopyridine based HPLC systems 1898:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 10 12 from differences 1265:1275 arg1 viability 1280:1288 viability 1280:1288 viability 1280:1288 However, cells did not display significant differences in viability and capabilities to migrate through an endothelial layer. 24726881 3 13 theme beta-1,4-mannosyl-glycoprotein 342:371 arg1 EC 413:414 EC 413:414 EC = 2.4.1.144 413:426 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 13 theme beta-1,4-mannosyl-glycoprotein 342:371 arg1 4-beta-N-acetylglucosaminyltransferase 373:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase 336:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) 336:436 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 0 14 theme melanoma 70:77 arg1 cells 79:83 human melanoma cells 64:83 human melanoma cells 64:83 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 2 15 theme exact 259:263 arg1 mechanisms 265:274 the exact mechanisms 255:274 the exact mechanisms 255:274 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 14 16 theme good 1877:1880 arg1 alternative 1882:1892 a good alternative 1875:1892 a good alternative for 2-aminopyridine based HPLC systems 1875:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 14 16 theme good 1877:1880 arg1 modification 1804:1815 The proposed modification 1791:1815 The proposed modification of the glycosyltransferase activity assay 1791:1857 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 13 17 theme better 1733:1738 arg1 understanding 1740:1752 better understanding 1733:1752 better understanding of their role in cancer progression 1733:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 5 18 theme functional 748:757 arg1 characterization 759:774 a preliminary functional characterization 734:774 a preliminary functional characterization 734:774 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 9 19 theme " 1118:1118 arg1 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 19 theme " 1118:1118 arg1 amounts 1098:1104 Increased amounts 1088:1104 Increased amounts of "bisected" and beta1-6 branched N-glycans 1088:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 19 theme " 1118:1118 arg1 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 13 20 from role 1763:1766 arg1 progression 1778:1788 cancer progression 1771:1788 cancer progression 1771:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 5 21 theme melanoma 651:658 arg1 lines 665:669 two human melanoma cell lines 641:669 two human melanoma cell lines with induced overexpression of GnT-III or GnT-V 641:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 3 22 theme = 416:416 arg1 EC 413:414 EC 413:414 EC = 2.4.1.144 413:426 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 22 theme = 416:416 arg1 4-beta-N-acetylglucosaminyltransferase 373:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase 336:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) 336:436 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 0 23 from Overexpression 0:13 arg1 cells 79:83 human melanoma cells 64:83 human melanoma cells 64:83 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 3 24 gly alpha-1,6-mannosylglycoprotein 442:471 arg1 alpha-1,6-mannosylglycoprotein 442:471 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A 442:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) 442:536 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 12 25 theme WM-266-4-GnT-III 1564:1579 arg1 cells 1581:1585 WM-266-4-GnT-III cells 1564:1585 WM-266-4-GnT-III cells 1564:1585 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 6 26 theme WM-266-4 777:784 arg1 cells 786:790 WM-266-4 cells 777:790 WM-266-4 cells 777:790 WM-266-4 cells were stably transfected with human MGAT3 or MGAT5 cDNAs. 24726881 9 27 dep molecule 1190:1197 arg1 known 1200:1204 known 1200:1204 known as MCAM/MUC18 1200:1218 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 14 28 theme assay 1853:1857 arg1 alternative 1882:1892 a good alternative 1875:1892 a good alternative for 2-aminopyridine based HPLC systems 1875:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 14 28 theme assay 1853:1857 arg1 modification 1804:1815 The proposed modification 1791:1815 The proposed modification of the glycosyltransferase activity assay 1791:1857 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 13 29 theme branching 1719:1727 arg1 understanding 1740:1752 better understanding 1733:1752 better understanding of their role in cancer progression 1733:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 13 29 theme branching 1719:1727 arg1 mechanisms 1695:1704 mechanisms 1695:1704 mechanisms of N-glycans branching 1695:1727 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 12 30 from glycosyltransferases 1540:1559 arg1 cells 1581:1585 WM-266-4-GnT-III cells 1564:1585 WM-266-4-GnT-III cells 1564:1585 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 1 31 theme MCAM 103:106 arg1 N-glycosylation 108:122 MCAM N-glycosylation 103:122 MCAM N-glycosylation 103:122 Implications for MCAM N-glycosylation. 24726881 9 32 theme melanoma 1167:1174 arg1 molecule 1190:1197 melanoma cell adhesion molecule 1167:1197 melanoma cell adhesion molecule (known as MCAM/MUC18) 1167:1219 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 0 33 theme N-acetylglucosaminyltransferases 18:49 arg1 Overexpression 0:13 Overexpression 0:13 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells 0:83 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 7 34 theme GnT-V 861:865 arg1 activities 867:876 GnT-III and GnT-V activities 849:876 GnT-III and GnT-V activities 849:876 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 9 35 theme adhesion 1181:1188 arg1 molecule 1190:1197 melanoma cell adhesion molecule 1167:1197 melanoma cell adhesion molecule (known as MCAM/MUC18) 1167:1219 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 7 36 theme GnT-III 849:855 arg1 activities 867:876 GnT-III and GnT-V activities 849:876 GnT-III and GnT-V activities 849:876 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 3 37 theme alpha-1,6-mannosylglycoprotein 442:471 arg1 EC 515:516 EC = 2.4.1.155 515:528 EC = 2.4.1.155 515:528 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 37 theme alpha-1,6-mannosylglycoprotein 442:471 arg1 A 512:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A 442:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) 442:536 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 14 38 theme activity 1844:1851 arg1 assay 1853:1857 the glycosyltransferase activity assay 1820:1857 the glycosyltransferase activity assay 1820:1857 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 13 39 theme obtained 1653:1660 arg1 model 1662:1666 The obtained model 1649:1666 The obtained model 1649:1666 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 13 39 theme obtained 1653:1660 arg1 useful 1675:1680 useful 1675:1680 useful 1675:1680 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 3 40 gly beta-1,4-mannosyl-glycoprotein 342:371 arg1 beta-1,4-mannosyl-glycoprotein 342:371 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase 336:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) 336:436 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 41 dep = 416:416 arg1 2.4.1.144 418:426 2.4.1.144 418:426 2.4.1.144 418:426 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 6 42 theme human 821:825 arg1 MGAT3 827:831 human MGAT3 or MGAT5 cDNAs 821:846 MGAT3 827:831 WM-266-4 cells were stably transfected with human MGAT3 or MGAT5 cDNAs. 24726881 0 43 dep N-acetylglucosaminyltransferases 18:49 arg1 N-acetylglucosaminyltransferases 18:49 N-acetylglucosaminyltransferases III and V 18:59 N-acetylglucosaminyltransferases III and V 18:59 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 0 43 dep N-acetylglucosaminyltransferases 18:49 arg1 V 59:59 V 59:59 V 59:59 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 0 43 dep N-acetylglucosaminyltransferases 18:49 arg1 III 51:53 III 51:53 III 51:53 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 2 44 theme bisecting 199:207 arg1 N-acetylglucosamine 210:228 "bisecting" N-acetylglucosamine 198:228 "bisecting" N-acetylglucosamine 198:228 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 9 45 theme bisected 1110:1117 arg1 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 14 46 theme glycosyltransferase 1824:1842 arg1 assay 1853:1857 the glycosyltransferase activity assay 1820:1857 the glycosyltransferase activity assay 1820:1857 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 11 47 theme " 1444:1444 arg1 N-glycans 1446:1454 "bisected" N-glycans 1435:1454 "bisected" N-glycans 1435:1454 To the best of our knowledge, the result of our study is the first to demonstrate that "bisected" N-glycans can be carried by MCAM. 24726881 7 48 theme N-glycan 937:944 arg1 acceptor 946:953 N-glycan acceptor 937:953 N-glycan acceptor 937:953 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 10 49 theme significant 1253:1263 arg1 differences 1265:1275 significant differences 1253:1275 significant differences in viability 1253:1288 However, cells did not display significant differences in viability and capabilities to migrate through an endothelial layer. 24726881 5 50 theme induced 676:682 arg1 overexpression 684:697 induced overexpression 676:697 induced overexpression of GnT-III or GnT-V 676:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 10 51 from capabilities 1294:1305 arg1 viability 1280:1288 viability 1280:1288 viability 1280:1288 However, cells did not display significant differences in viability and capabilities to migrate through an endothelial layer. 24726881 2 52 theme cancer 146:151 arg1 pathogenesis 153:164 cancer pathogenesis 146:164 cancer pathogenesis 146:164 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 12 53 theme enzyme 1641:1646 arg1 overexpression 1614:1627 the overexpression 1610:1627 the overexpression of only one enzyme 1610:1646 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 9 54 attach present 1156:1162 arg1 molecule 1190:1197 melanoma cell adhesion molecule 1167:1197 melanoma cell adhesion molecule (known as MCAM/MUC18) 1167:1219 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 54 attach present 1156:1162 arg2 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 54 attach present 1156:1162 arg2 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 54 attach present 1156:1162 arg2 amounts 1098:1104 Increased amounts 1088:1104 Increased amounts of "bisected" and beta1-6 branched N-glycans 1088:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 2 55 with branches 242:249 arg1 N-acetylglucosamine 210:228 "bisecting" N-acetylglucosamine 198:228 "bisecting" N-acetylglucosamine 198:228 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 7 56 theme HPLC 904:907 arg1 method 909:914 a novel HPLC method 896:914 a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989) 896:1013 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 2 57 from role 138:141 arg1 pathogenesis 153:164 cancer pathogenesis 146:164 cancer pathogenesis 146:164 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 13 58 theme role 1763:1766 arg1 understanding 1740:1752 better understanding 1733:1752 better understanding of their role in cancer progression 1733:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 13 58 theme role 1763:1766 arg1 mechanisms 1695:1704 mechanisms 1695:1704 mechanisms of N-glycans branching 1695:1727 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 12 59 theme overexpression 1614:1627 arg1 consequence 1595:1605 the consequence 1591:1605 the consequence of the overexpression of only one enzyme 1591:1646 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 12 59 theme overexpression 1614:1627 arg1 modification 1500:1511 increased modification 1490:1511 increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells 1490:1585 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 13 60 theme cancer 1771:1776 arg1 progression 1778:1788 cancer progression 1771:1788 cancer progression 1771:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 14 61 theme HPLC 1920:1923 arg1 systems 1925:1931 2-aminopyridine based HPLC systems 1898:1931 2-aminopyridine based HPLC systems 1898:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 9 62 theme Increased 1088:1096 arg1 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 62 theme Increased 1088:1096 arg1 amounts 1098:1104 Increased amounts 1088:1104 Increased amounts of "bisected" and beta1-6 branched N-glycans 1088:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 62 theme Increased 1088:1096 arg1 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 5 63 theme preliminary 736:746 arg1 characterization 759:774 a preliminary functional characterization 734:774 a preliminary functional characterization 734:774 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 14 64 theme 2-aminopyridine 1898:1912 arg1 systems 1925:1931 2-aminopyridine based HPLC systems 1898:1931 2-aminopyridine based HPLC systems 1898:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 10 65 theme endothelial 1329:1339 arg1 layer 1341:1345 an endothelial layer 1326:1345 an endothelial layer 1326:1345 However, cells did not display significant differences in viability and capabilities to migrate through an endothelial layer. 24726881 13 66 from progression 1778:1788 arg1 understanding 1740:1752 better understanding 1733:1752 better understanding of their role in cancer progression 1733:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 13 66 from progression 1778:1788 arg1 mechanisms 1695:1704 mechanisms 1695:1704 mechanisms of N-glycans branching 1695:1727 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 3 67 theme Golgi 336:340 arg1 EC 413:414 EC 413:414 EC = 2.4.1.144 413:426 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 67 theme Golgi 336:340 arg1 4-beta-N-acetylglucosaminyltransferase 373:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase 336:410 Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) 336:436 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 4 68 theme MGAT5 589:593 arg1 genes 595:599 MGAT5 genes 589:599 MGAT5 genes 589:599 The enzymes are encoded by MGAT3 and MGAT5 genes. 24726881 5 69 theme cell 660:663 arg1 lines 665:669 two human melanoma cell lines 641:669 two human melanoma cell lines with induced overexpression of GnT-III or GnT-V 641:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 9 70 theme branched 1132:1139 arg1 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 13 71 theme N-glycans 1709:1717 arg1 branching 1719:1727 N-glycans branching 1709:1727 N-glycans branching 1709:1727 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 3 72 theme = 518:518 arg1 EC 515:516 EC = 2.4.1.155 515:528 EC = 2.4.1.155 515:528 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 72 theme = 518:518 arg1 A 512:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A 442:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) 442:536 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 5 73 theme human 645:649 arg1 lines 665:669 two human melanoma cell lines 641:669 two human melanoma cell lines with induced overexpression of GnT-III or GnT-V 641:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 9 74 theme cell 1176:1179 arg1 molecule 1190:1197 melanoma cell adhesion molecule 1167:1197 melanoma cell adhesion molecule (known as MCAM/MUC18) 1167:1219 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 75 theme N-glycans 1141:1149 arg1 " 1118:1118 "bisected" 1109:1118 "bisected" 1109:1118 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 75 theme N-glycans 1141:1149 arg1 amounts 1098:1104 Increased amounts 1088:1104 Increased amounts of "bisected" and beta1-6 branched N-glycans 1088:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 9 75 theme N-glycans 1141:1149 arg1 N-glycans 1141:1149 beta1-6 branched N-glycans 1124:1149 beta1-6 branched N-glycans 1124:1149 Increased amounts of "bisected" and beta1-6 branched N-glycans were present on melanoma cell adhesion molecule (known as MCAM/MUC18). 24726881 12 76 theme protein 1521:1527 arg1 consequence 1595:1605 the consequence 1591:1605 the consequence of the overexpression of only one enzyme 1591:1646 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 12 76 theme protein 1521:1527 arg1 modification 1500:1511 increased modification 1490:1511 increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells 1490:1585 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 3 77 dep EC 515:516 arg1 GnT-V 531:535 GnT-V 531:535 GnT-V 531:535 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 6 78 theme MGAT5 836:840 arg1 cDNAs 842:846 human MGAT3 or MGAT5 cDNAs 821:846 cDNAs 842:846 WM-266-4 cells were stably transfected with human MGAT3 or MGAT5 cDNAs. 24726881 0 79 theme human 64:68 arg1 cells 79:83 human melanoma cells 64:83 human melanoma cells 64:83 Overexpression of N-acetylglucosaminyltransferases III and V in human melanoma cells. 24726881 5 80 with lines 665:669 arg1 overexpression 684:697 induced overexpression 676:697 induced overexpression of GnT-III or GnT-V 676:717 The aim of this study was to establish two human melanoma cell lines with induced overexpression of GnT-III or GnT-V and to perform a preliminary functional characterization. 24726881 2 81 with N-glycans 183:191 arg1 N-acetylglucosamine 210:228 "bisecting" N-acetylglucosamine 198:228 "bisecting" N-acetylglucosamine 198:228 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 3 82 theme 6-beta-N-acetylglucosaminyltransferase 473:510 arg1 EC 515:516 EC = 2.4.1.155 515:528 EC = 2.4.1.155 515:528 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 82 theme 6-beta-N-acetylglucosaminyltransferase 473:510 arg1 A 512:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A 442:512 alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) 442:536 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 3 83 dep EC 413:414 arg1 GnT-III 429:435 GnT-III 429:435 GnT-III 429:435 Two structures are formed by Golgi beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase (EC = 2.4.1.144, GnT-III) and alpha-1,6-mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A (EC = 2.4.1.155, GnT-V) respectively. 24726881 13 84 from understanding 1740:1752 arg1 progression 1778:1788 cancer progression 1771:1788 cancer progression 1771:1788 The obtained model can be useful for studying mechanisms of N-glycans branching and better understanding of their role in cancer progression. 24726881 12 85 theme increased 1490:1498 arg1 consequence 1595:1605 the consequence 1591:1605 the consequence of the overexpression of only one enzyme 1591:1646 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 12 85 theme increased 1490:1498 arg1 modification 1500:1511 increased modification 1490:1511 increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells 1490:1585 Moreover, increased modification of this protein by the two glycosyltransferases in WM-266-4-GnT-III cells was the consequence of the overexpression of only one enzyme. 24726881 2 86 theme " 208:208 arg1 N-acetylglucosamine 210:228 "bisecting" N-acetylglucosamine 198:228 "bisecting" N-acetylglucosamine 198:228 An important role in cancer pathogenesis is attributed to N-glycans with "bisecting" N-acetylglucosamine and beta1-6 branches but the exact mechanisms still remain to be elucidated. 24726881 14 87 theme proposed 1795:1802 arg1 alternative 1882:1892 a good alternative 1875:1892 a good alternative for 2-aminopyridine based HPLC systems 1875:1931 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 14 87 theme proposed 1795:1802 arg1 modification 1804:1815 The proposed modification 1791:1815 The proposed modification of the glycosyltransferase activity assay 1791:1857 The proposed modification of the glycosyltransferase activity assay has shown to be a good alternative for 2-aminopyridine based HPLC systems. 24726881 7 88 with labeling 925:932 arg1 2-aminobenzamide 960:975 2-aminobenzamide 960:975 2-aminobenzamide (adapted from Taniguchi et al., 1989) 960:1013 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 24726881 7 89 theme acceptor 946:953 arg1 labeling 925:932 labeling 925:932 labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989) 925:1013 GnT-III and GnT-V activities were assayed with a novel HPLC method based on labeling of N-glycan acceptor with 2-aminobenzamide (adapted from Taniguchi et al., 1989). 26075384 6 0 dep stages 895:900 arg1 benign 903:908 benign 903:908 benign 903:908 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 9 1 theme potential 1424:1432 arg1 carcinomas 1434:1443 mucinous low malignant potential carcinomas 1401:1443 mucinous low malignant potential carcinomas (LMPs, borderline) 1401:1462 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 6 2 theme different 885:893 arg1 stages 895:900 different stages 885:900 different stages (benign, borderline, malignant) 885:932 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 6 3 dep benign 903:908 arg1 malignant 923:931 malignant 923:931 malignant 923:931 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 6 3 dep benign 903:908 arg1 borderline 911:920 borderline 911:920 borderline 911:920 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 7 4 theme Mass 1107:1110 arg1 ESI-MSn 1126:1132 ESI-MSn 1126:1132 ESI-MSn 1126:1132 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 4 theme Mass 1107:1110 arg1 Spectrometry 1112:1123 Electrospray Ionization tandem Mass Spectrometry 1076:1123 Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1076:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 13 5 theme high-grade 2162:2171 arg1 carcinomas 2182:2191 serous and high-grade mucinous carcinomas 2151:2191 serous and high-grade mucinous carcinomas based on their O-glycan profiles 2151:2224 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 8 6 theme oligosaccharides 1158:1173 arg1 LC-ESI-MSn 1140:1149 The LC-ESI-MSn 1136:1149 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids 1136:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 8 7 theme fucose 1239:1244 arg1 structures 1257:1266 fucose containing structures 1239:1266 fucose containing structures such as blood group ABO antigens 1239:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 8 7 theme fucose 1239:1244 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 7 8 theme Ionization 1089:1098 arg1 ESI-MSn 1126:1132 ESI-MSn 1126:1132 ESI-MSn 1126:1132 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 8 theme Ionization 1089:1098 arg1 Spectrometry 1112:1123 Electrospray Ionization tandem Mass Spectrometry 1076:1123 Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1076:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 2 9 theme epithelial 265:274 arg1 cells 284:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 8 10 theme ovarian 1180:1186 arg1 fluids 1193:1198 ovarian cyst fluids 1180:1198 ovarian cyst fluids 1180:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 6 11 theme mucinous 858:865 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 8 12 theme structures 1257:1266 arg1 expression 1225:1234 expression 1225:1234 expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes 1225:1323 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 10 13 theme fucosylated 1583:1593 arg1 structures 1595:1604 fucosylated structures 1583:1604 fucosylated structures 1583:1604 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 2 14 theme secreted 305:312 arg1 mucins 314:319 secreted mucins 305:319 secreted mucins 305:319 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 6 15 theme grades 938:943 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 4 16 theme effective 614:622 arg1 biomarkers 624:633 more effective biomarkers 609:633 more effective biomarkers for ovarian cancer 609:652 The study of these proteins can potentially lead to the identification of more effective biomarkers for ovarian cancer. 26075384 11 17 theme group 1767:1771 arg1 antigens 1773:1780 blood group antigens 1761:1780 blood group antigens 1761:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 0 18 theme Ovarian 112:118 arg1 Tumors 120:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 9 19 theme mucinous 1468:1475 arg1 carcinomas 1487:1496 mucinous low-grade carcinomas 1468:1496 mucinous low-grade carcinomas 1468:1496 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 8 20 theme group 1282:1286 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 13 21 theme serous 2151:2156 arg1 carcinomas 2182:2191 serous and high-grade mucinous carcinomas 2151:2191 serous and high-grade mucinous carcinomas based on their O-glycan profiles 2151:2224 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 1 22 gly O-glycosylated 158:171 arg1 proteins 173:180 heavily O-glycosylated proteins 150:180 heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer 150:255 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 1 22 gly O-glycosylated 158:171 arg1 Mucins 139:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 7 23 theme Liquid 1038:1043 arg1 LC 1061:1062 LC 1061:1062 LC 1061:1062 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 23 theme Liquid 1038:1043 arg1 Chromatography 1045:1058 negative-ion graphitized carbon Liquid Chromatography 1006:1058 negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1006:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 8 24 theme Lewis-type 1305:1314 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 8 24 theme Lewis-type 1305:1314 arg1 epitopes 1316:1323 Lewis-type epitopes 1305:1323 Lewis-type epitopes 1305:1323 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 11 25 theme high 1733:1736 arg1 level 1738:1742 a high level 1731:1742 a high level of expression of blood group antigens 1731:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 7 26 theme graphitized 1019:1029 arg1 LC 1061:1062 LC 1061:1062 LC 1061:1062 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 26 theme graphitized 1019:1029 arg1 Chromatography 1045:1058 negative-ion graphitized carbon Liquid Chromatography 1006:1058 negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1006:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 13 27 theme low-grade 2102:2110 arg1 carcinomas 2112:2121 mucinous low-grade carcinomas 2093:2121 mucinous low-grade carcinomas 2093:2121 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 1 28 theme BACKGROUND 128:137 arg1 proteins 173:180 heavily O-glycosylated proteins 150:180 heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer 150:255 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 1 28 theme BACKGROUND 128:137 arg1 Mucins 139:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 13 29 dep serous 2151:2156 arg1 mucinous 2173:2180 mucinous 2173:2180 mucinous 2173:2180 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 13 30 theme benign 2066:2071 arg1 CONCLUSION 2046:2055 CONCLUSION 2046:2055 CONCLUSION Mucinous benign 2046:2071 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 9 31 theme blood 1519:1523 arg1 groups 1525:1530 blood groups 1519:1530 blood groups 1519:1530 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 5 32 theme acidic 746:751 arg1 glycoproteins 753:765 acidic glycoproteins 746:765 acidic glycoproteins secreted into ovarian cyst fluids 746:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 12 33 theme ovarian 1902:1908 arg1 tumors 1910:1915 serous and mucinous ovarian tumors 1882:1915 serous and mucinous ovarian tumors 1882:1915 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 1 34 theme O-glycosylated 158:171 arg1 proteins 173:180 heavily O-glycosylated proteins 150:180 heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer 150:255 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 1 34 theme O-glycosylated 158:171 arg1 Mucins 139:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins 128:144 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 5 35 gly O-glycosylation 727:741 arg1 study 671:675 this study 666:675 this study 666:675 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 5 35 gly O-glycosylation 727:741 arg1 MUC5AC 712:717 the MUC5AC 708:717 the MUC5AC 708:717 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 5 35 gly O-glycosylation 727:741 arg1 glycoproteins 753:765 acidic glycoproteins 746:765 acidic glycoproteins secreted into ovarian cyst fluids 746:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 0 36 theme Mucinous 81:88 arg1 Tumors 120:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 9 37 theme type 1542:1545 arg1 epitopes 1547:1554 Lewis type epitopes 1536:1554 Lewis type epitopes 1536:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 10 38 theme serous 1668:1673 arg1 carcinomas 1675:1684 serous carcinomas 1668:1684 serous carcinomas 1668:1684 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 2 39 theme ovarian 396:402 arg1 cancer 404:409 epithelial ovarian cancer 385:409 epithelial ovarian cancer 385:409 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 0 40 theme Serous 94:99 arg1 Tumors 120:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 0 41 theme Group 31:35 arg1 Antigens 37:44 Blood Group Antigens 25:44 Blood Group Antigens 25:44 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 6 42 with patients 833:840 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 10 43 theme high-grade 1631:1640 arg1 carcinomas 1651:1660 the high-grade mucinous carcinomas 1627:1660 the high-grade mucinous carcinomas 1627:1660 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 9 44 theme obtained 1338:1345 arg1 data 1347:1350 The obtained data 1334:1350 The obtained data 1334:1350 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 2 45 theme tumor 429:433 arg1 formation 435:443 tumor formation 429:443 tumor formation 429:443 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 12 46 dep spectrometry 2004:2015 arg1 compositions 2025:2036 average compositions 2017:2036 mass spectrometry average compositions (MSAC) 1999:2043 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 12 46 dep spectrometry 2004:2015 arg1 MSAC 2039:2042 MSAC 2039:2042 MSAC 2039:2042 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 0 47 theme Glycome 13:19 arg1 ABO 46:48 The O-Linked Glycome and Blood Group Antigens ABO 0:48 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 0:126 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 0 48 theme Blood 25:29 arg1 Antigens 37:44 Blood Group Antigens 25:44 Blood Group Antigens 25:44 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 9 49 theme serous 1364:1369 arg1 adenomas 1391:1398 benign adenomas 1384:1398 benign adenomas 1384:1398 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 12 50 theme mass 1999:2002 arg1 spectrometry 2004:2015 mass spectrometry average compositions (MSAC) 1999:2043 mass spectrometry average compositions (MSAC) 1999:2043 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 8 51 from differences 1210:1220 arg1 expression 1225:1234 expression 1225:1234 expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes 1225:1323 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 2 52 theme abnormal 332:339 arg1 expression 341:350 their abnormal expression 326:350 their abnormal expression 326:350 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 5 53 theme ovarian 781:787 arg1 fluids 794:799 ovarian cyst fluids 781:799 ovarian cyst fluids 781:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 9 54 theme mucinous 1375:1382 arg1 adenomas 1391:1398 benign adenomas 1384:1398 benign adenomas 1384:1398 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 10 55 from contrast 1560:1567 arg1 low 1611:1613 low 1611:1613 low 1611:1613 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 10 56 from carcinomas 1651:1660 arg1 abundant 1615:1622 abundant 1615:1622 abundant 1615:1622 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 3 57 theme acidic 513:518 arg1 glycoproteins 520:532 acidic glycoproteins 513:532 acidic glycoproteins 513:532 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 3 58 gly glycoproteins 520:532 arg1 glycoproteins 520:532 acidic glycoproteins 513:532 acidic glycoproteins 513:532 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 11 59 theme ovarian 1704:1710 arg1 tumors 1712:1717 the ovarian tumors 1700:1717 the ovarian tumors that showed a high level of expression of blood group antigens 1700:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 3 60 theme rich 497:500 arg1 fluids 471:476 The cyst fluids 462:476 The cyst fluids 462:476 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 3 60 theme rich 497:500 arg1 source 502:507 a rich source 495:507 a rich source for acidic glycoproteins 495:532 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 9 61 theme mucinous 1401:1408 arg1 carcinomas 1434:1443 mucinous low malignant potential carcinomas 1401:1443 mucinous low malignant potential carcinomas (LMPs, borderline) 1401:1462 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 12 62 theme average 2017:2023 arg1 compositions 2025:2036 average compositions 2017:2036 mass spectrometry average compositions (MSAC) 1999:2043 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 12 62 theme average 2017:2023 arg1 MSAC 2039:2042 MSAC 2039:2042 MSAC 2039:2042 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 8 63 from fluids 1193:1198 arg1 LC-ESI-MSn 1140:1149 The LC-ESI-MSn 1136:1149 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids 1136:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 8 63 from fluids 1193:1198 arg1 oligosaccharides 1158:1173 the oligosaccharides 1154:1173 the oligosaccharides from ovarian cyst fluids 1154:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 12 64 theme cluster 1962:1968 arg1 analysis 1970:1977 a hierarchical cluster analysis 1947:1977 a hierarchical cluster analysis 1947:1977 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 10 65 from abundant 1615:1622 arg1 carcinomas 1651:1660 the high-grade mucinous carcinomas 1627:1660 the high-grade mucinous carcinomas 1627:1660 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 9 66 theme malignant 1414:1422 arg1 carcinomas 1434:1443 mucinous low malignant potential carcinomas 1401:1443 mucinous low malignant potential carcinomas (LMPs, borderline) 1401:1462 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 6 67 theme stages 895:900 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 4 68 theme proteins 554:561 arg1 study 539:543 The study 535:543 The study of these proteins 535:561 The study of these proteins can potentially lead to the identification of more effective biomarkers for ovarian cancer. 26075384 7 69 theme tandem 1100:1105 arg1 ESI-MSn 1126:1132 ESI-MSn 1126:1132 ESI-MSn 1126:1132 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 69 theme tandem 1100:1105 arg1 Spectrometry 1112:1123 Electrospray Ionization tandem Mass Spectrometry 1076:1123 Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1076:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 10 70 from low 1611:1613 arg1 contrast 1560:1567 contrast 1560:1567 contrast 1560:1567 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 7 71 theme Electrospray 1076:1087 arg1 ESI-MSn 1126:1132 ESI-MSn 1126:1132 ESI-MSn 1126:1132 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 71 theme Electrospray 1076:1087 arg1 Spectrometry 1112:1123 Electrospray Ionization tandem Mass Spectrometry 1076:1123 Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1076:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 2 72 theme ovarian 276:282 arg1 cells 284:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 0 73 theme Antigens 37:44 arg1 ABO 46:48 The O-Linked Glycome and Blood Group Antigens ABO 0:48 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 0:126 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 6 74 theme ovarian 867:873 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 4 75 theme ovarian 639:645 arg1 cancer 647:652 ovarian cancer 639:652 ovarian cancer 639:652 The study of these proteins can potentially lead to the identification of more effective biomarkers for ovarian cancer. 26075384 10 76 theme structures 1595:1604 arg1 type 1575:1578 this type 1570:1578 this type of fucosylated structures 1570:1604 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 12 77 theme hierarchical 1949:1960 arg1 analysis 1970:1977 a hierarchical cluster analysis 1947:1977 a hierarchical cluster analysis 1947:1977 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 8 78 theme containing 1246:1255 arg1 structures 1257:1266 fucose containing structures 1239:1266 fucose containing structures such as blood group ABO antigens 1239:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 8 78 theme containing 1246:1255 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 7 79 link O-linked 950:957 arg1 oligosaccharides 959:974 The O-linked oligosaccharides 946:974 The O-linked oligosaccharides 946:974 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 8 80 theme cyst 1188:1191 arg1 fluids 1193:1198 ovarian cyst fluids 1180:1198 ovarian cyst fluids 1180:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 11 81 theme antigens 1773:1780 arg1 expression 1747:1756 expression 1747:1756 expression of blood group antigens 1747:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 9 82 theme low-grade 1477:1485 arg1 carcinomas 1487:1496 mucinous low-grade carcinomas 1468:1496 mucinous low-grade carcinomas 1468:1496 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 4 83 theme biomarkers 624:633 arg1 identification 591:604 the identification 587:604 the identification of more effective biomarkers for ovarian cancer 587:652 The study of these proteins can potentially lead to the identification of more effective biomarkers for ovarian cancer. 26075384 7 84 theme carbon 1031:1036 arg1 LC 1061:1062 LC 1061:1062 LC 1061:1062 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 84 theme carbon 1031:1036 arg1 Chromatography 1045:1058 negative-ion graphitized carbon Liquid Chromatography 1006:1058 negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1006:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 9 85 dep carcinomas 1434:1443 arg1 borderline 1452:1461 borderline 1452:1461 borderline 1452:1461 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 9 85 dep carcinomas 1434:1443 arg1 LMPs 1446:1449 LMPs 1446:1449 LMPs 1446:1449 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 8 86 theme ABO 1288:1290 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 11 87 theme strong 1798:1803 arg1 reactivity 1805:1814 a strong reactivity 1796:1814 a strong reactivity towards the MUC5AC antibody 1796:1842 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 7 88 theme negative-ion 1006:1017 arg1 LC 1061:1062 LC 1061:1062 LC 1061:1062 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 7 88 theme negative-ion 1006:1017 arg1 Chromatography 1045:1058 negative-ion graphitized carbon Liquid Chromatography 1006:1058 negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn) 1006:1133 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 13 89 theme mucinous 2093:2100 arg1 carcinomas 2112:2121 mucinous low-grade carcinomas 2093:2121 mucinous low-grade carcinomas 2093:2121 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 2 90 theme Normal 258:263 arg1 cells 284:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells 258:288 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 9 91 theme high 1505:1508 arg1 level 1510:1514 a high level 1503:1514 a high level of blood groups and Lewis type epitopes 1503:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 0 92 theme Mucin-Type 53:62 arg1 Glycoproteins 64:76 Mucin-Type Glycoproteins 53:76 Mucin-Type Glycoproteins 53:76 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 8 93 theme epitopes 1316:1323 arg1 expression 1225:1234 expression 1225:1234 expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes 1225:1323 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 7 94 theme O-linked 950:957 arg1 oligosaccharides 959:974 The O-linked oligosaccharides 946:974 The O-linked oligosaccharides 946:974 The O-linked oligosaccharides were released and analyzed by negative-ion graphitized carbon Liquid Chromatography (LC) coupled to Electrospray Ionization tandem Mass Spectrometry (ESI-MSn). 26075384 9 95 theme groups 1525:1530 arg1 level 1510:1514 a high level 1503:1514 a high level of blood groups and Lewis type epitopes 1503:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 11 96 theme blood 1761:1765 arg1 antigens 1773:1780 blood group antigens 1761:1780 blood group antigens 1761:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 9 97 theme Lewis 1536:1540 arg1 epitopes 1547:1554 Lewis type epitopes 1536:1554 Lewis type epitopes 1536:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 0 98 from ABO 46:48 arg1 Glycoproteins 64:76 Mucin-Type Glycoproteins 53:76 Mucin-Type Glycoproteins 53:76 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 0 98 from ABO 46:48 arg1 Tumors 120:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 11 99 theme expression 1747:1756 arg1 level 1738:1742 a high level 1731:1742 a high level of expression of blood group antigens 1731:1780 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 0 100 theme Epithelial 101:110 arg1 Tumors 120:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 Mucinous and Serous Epithelial Ovarian Tumors 81:125 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 12 101 theme serous 1882:1887 arg1 tumors 1910:1915 serous and mucinous ovarian tumors 1882:1915 serous and mucinous ovarian tumors 1882:1915 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 9 102 theme epitopes 1547:1554 arg1 level 1510:1514 a high level 1503:1514 a high level of blood groups and Lewis type epitopes 1503:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 10 103 gly fucosylated 1583:1593 arg1 structures 1595:1604 fucosylated structures 1583:1604 fucosylated structures 1583:1604 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 2 104 theme epithelial 385:394 arg1 cancer 404:409 epithelial ovarian cancer 385:409 epithelial ovarian cancer 385:409 Normal epithelial ovarian cells do not express secreted mucins, but their abnormal expression has previously been described in epithelial ovarian cancer and may relate to tumor formation and progression. 26075384 5 105 theme cyst 789:792 arg1 fluids 794:799 ovarian cyst fluids 781:799 ovarian cyst fluids 781:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 12 106 theme mucinous 1893:1900 arg1 tumors 1910:1915 serous and mucinous ovarian tumors 1882:1915 serous and mucinous ovarian tumors 1882:1915 To visualize the differences between serous and mucinous ovarian tumors based on the O-glycosylation, a hierarchical cluster analysis was performed using mass spectrometry average compositions (MSAC). 26075384 5 107 dep METHODS 655:661 arg1 analyzed 681:688 analyzed 681:688 analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids 681:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 8 108 from LC-ESI-MSn 1140:1149 arg1 fluids 1193:1198 ovarian cyst fluids 1180:1198 ovarian cyst fluids 1180:1198 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 3 109 theme cyst 466:469 arg1 fluids 471:476 The cyst fluids 462:476 The cyst fluids 462:476 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 3 109 theme cyst 466:469 arg1 source 502:507 a rich source 495:507 a rich source for acidic glycoproteins 495:532 The cyst fluids were shown to be a rich source for acidic glycoproteins. 26075384 5 110 theme glycoproteins 753:765 arg1 O-glycosylation 727:741 the O-glycosylation 723:741 the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids 723:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 5 110 theme glycoproteins 753:765 arg1 expression 694:703 the expression 690:703 the expression of the MUC5AC 690:717 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 6 111 theme serous 847:852 arg1 tumors 875:880 serous and mucinous ovarian tumors 847:880 serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades 847:943 The samples were obtained from patients with serous and mucinous ovarian tumors of different stages (benign, borderline, malignant) and grades. 26075384 9 112 dep RESULTS 1326:1332 arg1 showed 1352:1357 showed 1352:1357 showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes 1352:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 9 113 contain have 1498:1501 arg2 level 1510:1514 a high level 1503:1514 a high level of blood groups and Lewis type epitopes 1503:1554 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 9 113 contain have 1498:1501 arg1 adenomas 1391:1398 benign adenomas 1384:1398 benign adenomas 1384:1398 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 9 113 contain have 1498:1501 arg1 carcinomas 1434:1443 mucinous low malignant potential carcinomas 1401:1443 mucinous low malignant potential carcinomas (LMPs, borderline) 1401:1462 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 9 113 contain have 1498:1501 arg1 carcinomas 1487:1496 mucinous low-grade carcinomas 1468:1496 mucinous low-grade carcinomas 1468:1496 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 1 114 theme important 232:240 arg1 role 242:245 an important role 229:245 an important role 229:245 BACKGROUND Mucins are heavily O-glycosylated proteins where the glycosylation has been shown to play an important role in cancer. 26075384 13 115 theme O-glycan 2208:2215 arg1 profiles 2217:2224 their O-glycan profiles 2202:2224 their O-glycan profiles 2202:2224 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 9 116 theme benign 1384:1389 arg1 adenomas 1391:1398 benign adenomas 1384:1398 benign adenomas 1384:1398 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 26075384 5 117 theme MUC5AC 712:717 arg1 O-glycosylation 727:741 the O-glycosylation 723:741 the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids 723:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 5 117 theme MUC5AC 712:717 arg1 expression 694:703 the expression 690:703 the expression of the MUC5AC 690:717 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 13 118 from carcinomas 2182:2191 arg1 different 2136:2144 different 2136:2144 different 2136:2144 CONCLUSION Mucinous benign and LMPs along with mucinous low-grade carcinomas appear to be different from serous and high-grade mucinous carcinomas based on their O-glycan profiles. 26075384 5 119 gly glycoproteins 753:765 arg1 glycoproteins 753:765 acidic glycoproteins 746:765 acidic glycoproteins secreted into ovarian cyst fluids 746:799 METHODS In this study, we analyzed the expression of the MUC5AC and the O-glycosylation of acidic glycoproteins secreted into ovarian cyst fluids. 26075384 11 120 theme MUC5AC 1828:1833 arg1 antibody 1835:1842 the MUC5AC antibody 1824:1842 the MUC5AC antibody 1824:1842 In addition, the ovarian tumors that showed a high level of expression of blood group antigens also revealed a strong reactivity towards the MUC5AC antibody. 26075384 8 121 theme blood 1276:1280 arg1 antigens 1292:1299 blood group ABO antigens 1276:1299 blood group ABO antigens 1276:1299 The LC-ESI-MSn of the oligosaccharides from ovarian cyst fluids displayed differences in expression of fucose containing structures such as blood group ABO antigens and Lewis-type epitopes. 26075384 10 122 theme mucinous 1642:1649 arg1 carcinomas 1651:1660 the high-grade mucinous carcinomas 1627:1660 the high-grade mucinous carcinomas 1627:1660 In contrast, this type of fucosylated structures were low abundant in the high-grade mucinous carcinomas or in serous carcinomas. 26075384 0 123 theme O-Linked 4:11 arg1 Glycome 13:19 O-Linked Glycome 4:19 O-Linked Glycome 4:19 The O-Linked Glycome and Blood Group Antigens ABO on Mucin-Type Glycoproteins in Mucinous and Serous Epithelial Ovarian Tumors. 26075384 9 124 theme low 1410:1412 arg1 carcinomas 1434:1443 mucinous low malignant potential carcinomas 1401:1443 mucinous low malignant potential carcinomas (LMPs, borderline) 1401:1462 RESULTS The obtained data showed that serous and mucinous benign adenomas, mucinous low malignant potential carcinomas (LMPs, borderline) and mucinous low-grade carcinomas have a high level of blood groups and Lewis type epitopes. 25369125 4 0 theme C-type 481:486 arg1 self-association 488:503 a C-type self-association 479:503 a C-type self-association 479:503 Two types of self-association have been described for this lectin: a C-type self-association and a N-type self-association. 25369125 7 1 theme affinity 896:903 arg1 variations 905:914 no significant specificity or affinity variations 866:914 variations 905:914 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 5 2 theme full 655:658 arg1 lectin 667:672 the full length lectin 651:672 the full length lectin 651:672 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 5 3 theme Light 599:603 arg1 Scattering 605:614 Dynamic Light Scattering 591:614 Dynamic Light Scattering using both the recombinant CRD and the full length lectin 591:672 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 7 4 theme significant 869:879 arg1 specificity 881:891 no significant specificity or affinity variations 866:914 specificity 881:891 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 3 5 theme quaternary 336:345 arg1 structure 347:355 the quaternary structure 332:355 the quaternary structure of Galectin-3 332:369 There are still uncertainties about the relationship between the quaternary structure of Galectin-3 and its carbohydrate binding properties. 25369125 5 6 theme length 660:665 arg1 lectin 667:672 the full length lectin 651:672 the full length lectin 651:672 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 9 7 theme N-terminal 1218:1227 arg1 interactions 1236:1247 N-terminal domain interactions 1218:1247 N-terminal domain interactions 1218:1247 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 6 8 theme N-type 712:717 arg1 self-association 719:734 N-type self-association 712:734 N-type self-association of full length Galectin-3 712:760 Our results proved that LNnT induces N-type self-association of full length Galectin-3. 25369125 1 9 from surface 133:139 arg1 nucleus 94:100 the nucleus 90:100 the nucleus 90:100 Human Galectin-3 is found in the nucleus, the cytoplasm and at the cell surface. 25369125 9 10 theme Galectin-3 1199:1208 arg1 self-association 1179:1194 the glycan-dependent self-association 1158:1194 the glycan-dependent self-association of Galectin-3 1158:1208 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 5 11 theme recombinant 631:641 arg1 CRD 643:645 the recombinant CRD 627:645 the recombinant CRD 627:645 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 5 12 theme Galectin-3 561:570 arg1 oligomerization 572:586 Galectin-3 oligomerization 561:586 Galectin-3 oligomerization 561:586 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 0 13 theme Glycan 0:5 arg1 dependence 7:16 Glycan dependence 0:16 Glycan dependence of Galectin-3 self-association properties 0:58 Glycan dependence of Galectin-3 self-association properties. 25369125 1 14 from cytoplasm 107:115 arg1 nucleus 94:100 the nucleus 90:100 the nucleus 90:100 Human Galectin-3 is found in the nucleus, the cytoplasm and at the cell surface. 25369125 7 15 theme Resonance 831:839 arg1 experiments 841:851 Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments 778:851 Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments 778:851 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 8 16 theme N-terminal 1039:1048 arg1 domain 1050:1055 the N-terminal domain 1035:1055 the N-terminal domain 1035:1055 NMR mapping clearly established that the N-terminal domain interacts with the CRD. 25369125 0 17 theme Galectin-3 21:30 arg1 properties 49:58 Galectin-3 self-association properties 21:58 Galectin-3 self-association properties 21:58 Glycan dependence of Galectin-3 self-association properties. 25369125 5 18 theme Dynamic 591:597 arg1 Scattering 605:614 Dynamic Light Scattering 591:614 Dynamic Light Scattering using both the recombinant CRD and the full length lectin 591:672 Herein, we have analyzed Galectin-3 oligomerization by Dynamic Light Scattering using both the recombinant CRD and the full length lectin. 25369125 7 19 theme domain 976:981 arg1 presence 949:956 the presence 945:956 the presence of the N-terminal domain of Galectin-3 945:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 3 20 theme Galectin-3 360:369 arg1 properties 400:409 its carbohydrate binding properties 375:409 its carbohydrate binding properties 375:409 There are still uncertainties about the relationship between the quaternary structure of Galectin-3 and its carbohydrate binding properties. 25369125 3 20 theme Galectin-3 360:369 arg1 structure 347:355 the quaternary structure 332:355 the quaternary structure of Galectin-3 332:369 There are still uncertainties about the relationship between the quaternary structure of Galectin-3 and its carbohydrate binding properties. 25369125 7 21 theme Surface 815:821 arg1 Resonance 831:839 Surface Plasmon Resonance 815:839 Surface Plasmon Resonance 815:839 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 0 22 theme properties 49:58 arg1 dependence 7:16 Glycan dependence 0:16 Glycan dependence of Galectin-3 self-association properties 0:58 Glycan dependence of Galectin-3 self-association properties. 25369125 7 23 theme N-terminal 965:974 arg1 Galectin-3 986:995 Galectin-3 986:995 Galectin-3 986:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 23 theme N-terminal 965:974 arg1 domain 976:981 the N-terminal domain 961:981 the N-terminal domain of Galectin-3 961:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 24 theme Plasmon 823:829 arg1 Resonance 831:839 Surface Plasmon Resonance 815:839 Surface Plasmon Resonance 815:839 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 0 25 theme self-association 32:47 arg1 properties 49:58 Galectin-3 self-association properties 21:58 Galectin-3 self-association properties 21:58 Glycan dependence of Galectin-3 self-association properties. 25369125 7 26 theme Magnetic 786:793 arg1 NMR 806:808 NMR 806:808 NMR 806:808 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 26 theme Magnetic 786:793 arg1 Resonance 795:803 Nuclear Magnetic Resonance 778:803 Nuclear Magnetic Resonance (NMR) 778:809 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 9 27 theme N-terminal 1127:1136 arg1 domain 1138:1143 the N-terminal domain 1123:1143 the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 1123:1208 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 9 28 theme glycan-dependent 1162:1177 arg1 self-association 1179:1194 the glycan-dependent self-association 1158:1194 the glycan-dependent self-association of Galectin-3 1158:1208 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 3 29 theme carbohydrate 379:390 arg1 properties 400:409 its carbohydrate binding properties 375:409 its carbohydrate binding properties 375:409 There are still uncertainties about the relationship between the quaternary structure of Galectin-3 and its carbohydrate binding properties. 25369125 4 30 theme N-type 511:516 arg1 self-association 518:533 a N-type self-association 509:533 a N-type self-association 509:533 Two types of self-association have been described for this lectin: a C-type self-association and a N-type self-association. 25369125 2 31 theme N-terminal 197:206 arg1 domain 208:213 an unfolded N-terminal domain 185:213 an unfolded N-terminal domain 185:213 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 31 theme N-terminal 197:206 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 9 32 theme domain 1138:1143 arg1 release 1112:1118 a release 1110:1118 a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 1110:1208 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 3 33 theme binding 392:398 arg1 properties 400:409 its carbohydrate binding properties 375:409 its carbohydrate binding properties 375:409 There are still uncertainties about the relationship between the quaternary structure of Galectin-3 and its carbohydrate binding properties. 25369125 2 34 theme unfolded 188:195 arg1 domain 208:213 an unfolded N-terminal domain 185:213 an unfolded N-terminal domain 185:213 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 34 theme unfolded 188:195 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 7 35 theme Nuclear 778:784 arg1 NMR 806:808 NMR 806:808 NMR 806:808 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 35 theme Nuclear 778:784 arg1 Resonance 795:803 Nuclear Magnetic Resonance 778:803 Nuclear Magnetic Resonance (NMR) 778:809 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 2 36 theme Recognition 245:255 arg1 CRD 265:267 CRD 265:267 CRD 265:267 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 36 theme Recognition 245:255 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 36 theme Recognition 245:255 arg1 Domain 257:262 a C-terminal Carbohydrate Recognition Domain 219:262 a C-terminal Carbohydrate Recognition Domain (CRD) 219:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 6 37 theme Galectin-3 751:760 arg1 self-association 719:734 N-type self-association 712:734 N-type self-association of full length Galectin-3 712:760 Our results proved that LNnT induces N-type self-association of full length Galectin-3. 25369125 8 38 theme NMR 998:1000 arg1 mapping 1002:1008 NMR mapping 998:1008 NMR mapping 998:1008 NMR mapping clearly established that the N-terminal domain interacts with the CRD. 25369125 7 39 theme related 934:940 arg1 carbohydrates 920:932 carbohydrates 920:932 carbohydrates related to the presence of the N-terminal domain of Galectin-3 920:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 2 40 dep domains 176:182 arg1 domain 208:213 an unfolded N-terminal domain 185:213 an unfolded N-terminal domain 185:213 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 40 dep domains 176:182 arg1 CRD 265:267 CRD 265:267 CRD 265:267 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 40 dep domains 176:182 arg1 Domain 257:262 a C-terminal Carbohydrate Recognition Domain 219:262 a C-terminal Carbohydrate Recognition Domain (CRD) 219:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 40 dep domains 176:182 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 4 41 theme self-association 425:440 arg1 types 416:420 Two types 412:420 Two types of self-association 412:440 Two types of self-association have been described for this lectin: a C-type self-association and a N-type self-association. 25369125 2 42 theme Carbohydrate 232:243 arg1 CRD 265:267 CRD 265:267 CRD 265:267 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 42 theme Carbohydrate 232:243 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 42 theme Carbohydrate 232:243 arg1 Domain 257:262 a C-terminal Carbohydrate Recognition Domain 219:262 a C-terminal Carbohydrate Recognition Domain (CRD) 219:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 6 43 theme length 744:749 arg1 Galectin-3 751:760 full length Galectin-3 739:760 full length Galectin-3 739:760 Our results proved that LNnT induces N-type self-association of full length Galectin-3. 25369125 9 44 theme domain 1229:1234 arg1 interactions 1236:1247 N-terminal domain interactions 1218:1247 N-terminal domain interactions 1218:1247 We propose that LNnT induces a release of the N-terminal domain resulting in the glycan-dependent self-association of Galectin-3 through N-terminal domain interactions. 25369125 2 45 theme C-terminal 221:230 arg1 CRD 265:267 CRD 265:267 CRD 265:267 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 45 theme C-terminal 221:230 arg1 domains 176:182 two domains 172:182 two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD) 172:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 2 45 theme C-terminal 221:230 arg1 Domain 257:262 a C-terminal Carbohydrate Recognition Domain 219:262 a C-terminal Carbohydrate Recognition Domain (CRD) 219:268 This lectin is constituted of two domains: an unfolded N-terminal domain and a C-terminal Carbohydrate Recognition Domain (CRD). 25369125 6 46 theme full 739:742 arg1 Galectin-3 751:760 full length Galectin-3 739:760 full length Galectin-3 739:760 Our results proved that LNnT induces N-type self-association of full length Galectin-3. 25369125 1 47 theme cell 128:131 arg1 surface 133:139 the cell surface 124:139 the cell surface 124:139 Human Galectin-3 is found in the nucleus, the cytoplasm and at the cell surface. 25369125 1 48 theme Human 61:65 arg1 Galectin-3 67:76 Human Galectin-3 61:76 Human Galectin-3 61:76 Human Galectin-3 is found in the nucleus, the cytoplasm and at the cell surface. 25369125 7 49 theme Galectin-3 986:995 arg1 Galectin-3 986:995 Galectin-3 986:995 Galectin-3 986:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 49 theme Galectin-3 986:995 arg1 domain 976:981 the N-terminal domain 961:981 the N-terminal domain of Galectin-3 961:995 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 25369125 7 50 theme Resonance 795:803 arg1 experiments 841:851 Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments 778:851 Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments 778:851 Moreover, from Nuclear Magnetic Resonance (NMR) and Surface Plasmon Resonance experiments, we observed no significant specificity or affinity variations for carbohydrates related to the presence of the N-terminal domain of Galectin-3. 28752594 6 0 theme Multi-parameter 890:904 arg1 models 906:911 Multi-parameter models 890:911 Multi-parameter models 890:911 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 7 1 theme triantennary 1114:1125 arg1 N-glycan 1127:1134 the bifucosylated triantennary N-glycan 1096:1134 the bifucosylated triantennary N-glycan (peak10, NA3F2) 1096:1150 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 5 2 theme glycoproteins 767:779 arg1 N-glycome 748:756 the N-glycome 744:756 the N-glycome of serum glycoproteins 744:779 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 6 3 theme antigen 974:980 arg1 19-9 982:985 carbohydrate antigen 19-9 961:985 carbohydrate antigen 19-9 961:985 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 1 4 theme biomarkers 256:265 arg1 diagnosis 134:142 the diagnosis 130:142 the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character 130:221 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 1 4 theme biomarkers 256:265 arg1 lack 227:230 lack 227:230 lack of effective diagnostic biomarkers 227:265 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 10 5 theme ECCA 1489:1492 arg1 sera 1481:1484 sera 1481:1484 sera of ECCA 1481:1492 In conclusion, sera of ECCA showed relatively specific N-glycome profiling patterns. 28752594 11 6 theme valuable 1596:1603 arg1 markers 1566:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 6 theme valuable 1596:1603 arg1 alternatives 1621:1632 novel, valuable and noninvasive alternatives 1589:1632 novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring 1589:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 6 theme valuable 1596:1603 arg1 models 1578:1583 models 1578:1583 models 1578:1583 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 6 7 used used 1015:1018 arg2 N-glycans 947:955 the N-glycans 943:955 the N-glycans 943:955 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 6 7 used used 1015:1018 arg2 CA19-9 988:993 CA19-9 988:993 CA19-9 988:993 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 6 7 used used 1015:1018 arg2 19-9 982:985 carbohydrate antigen 19-9 961:985 carbohydrate antigen 19-9 961:985 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 4 8 dep controls 633:640 arg1 n = 89 647:652 n = 89 647:652 n = 89 647:652 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 8 dep controls 633:640 arg1 HC 643:644 HC 643:644 HC 643:644 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 2 9 from component 346:354 arg1 progression 366:376 cancer progression 359:376 cancer progression 359:376 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 2 10 theme glycoproteins 311:323 arg1 N-glycosylation 292:306 N-glycosylation 292:306 N-glycosylation of glycoproteins 292:323 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 4 11 theme benign 581:586 arg1 ECCA 565:568 ECCA (n = 106) 565:578 ECCA (n = 106) 565:578 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 11 theme benign 581:586 arg1 disease 599:605 benign bile tract disease 581:605 benign bile tract disease (BBD, n = 60) 581:619 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 2 12 gly glycoproteins 311:323 arg1 glycoproteins 311:323 glycoproteins 311:323 glycoproteins 311:323 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 9 13 dep structures 1361:1370 arg1 NA3Fb 1380:1384 NA3Fb 1380:1384 NA3Fb 1380:1384 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 9 13 dep structures 1361:1370 arg1 NA4Fb 1395:1399 NA4Fb 1395:1399 NA4Fb 1395:1399 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 7 14 theme bifucosylated 1100:1112 arg1 N-glycan 1127:1134 the bifucosylated triantennary N-glycan 1096:1134 the bifucosylated triantennary N-glycan (peak10, NA3F2) 1096:1150 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 9 15 theme lymph 1426:1430 arg1 node 1432:1435 lymph node 1426:1435 lymph node metastasis in ECCA patients 1426:1463 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 5 16 theme DNA 797:799 arg1 technology 878:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 17 theme serum 684:688 arg1 2 μL 676:679 Only 2 μL 671:679 Only 2 μL of serum from individual patients 671:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 9 18 theme ECCA 1451:1454 arg1 patients 1456:1463 ECCA patients 1451:1463 ECCA patients 1451:1463 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 5 19 used used 719:722 arg2 2 μL 676:679 Only 2 μL 671:679 Only 2 μL of serum from individual patients 671:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 2 20 theme cancer 433:438 arg1 detection 440:448 cancer detection 433:448 cancer detection 433:448 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 1 21 theme Extensive 83:91 arg1 efforts 93:99 Extensive efforts 83:99 Extensive efforts 83:99 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 4 22 from controls 633:640 arg1 samples 538:544 255 serum samples 528:544 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) 528:653 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 2 23 theme molecular 409:417 arg1 component 346:354 a key component 340:354 a key component 340:354 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 2 23 theme molecular 409:417 arg1 signature 419:427 a distinct molecular signature 398:427 a distinct molecular signature for cancer detection 398:448 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 2 24 gly N-glycosylation 292:306 arg1 glycoproteins 311:323 glycoproteins 311:323 glycoproteins 311:323 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 5 25 gly glycoproteins 767:779 arg1 glycoproteins 767:779 serum glycoproteins 761:779 serum glycoproteins 761:779 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 4 26 with patients 551:558 arg1 disease 599:605 benign bile tract disease 581:605 benign bile tract disease (BBD, n = 60) 581:619 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 26 with patients 551:558 arg1 ECCA 565:568 ECCA (n = 106) 565:578 ECCA (n = 106) 565:578 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 9 27 from metastasis 1437:1446 arg1 patients 1456:1463 ECCA patients 1451:1463 ECCA patients 1451:1463 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 2 28 theme distinct 400:407 arg1 component 346:354 a key component 340:354 a key component 340:354 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 2 28 theme distinct 400:407 arg1 signature 419:427 a distinct molecular signature 398:427 a distinct molecular signature for cancer detection 398:448 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 11 29 theme ECCA 1637:1640 arg1 diagnosis 1642:1650 ECCA diagnosis 1637:1650 ECCA diagnosis 1637:1650 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 10 30 theme specific 1512:1519 arg1 patterns 1541:1548 relatively specific N-glycome profiling patterns 1501:1548 relatively specific N-glycome profiling patterns 1501:1548 In conclusion, sera of ECCA showed relatively specific N-glycome profiling patterns. 28752594 2 31 theme key 342:344 arg1 component 346:354 a key component 340:354 a key component 340:354 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 2 31 theme key 342:344 arg1 signature 419:427 a distinct molecular signature 398:427 a distinct molecular signature for cancer detection 398:448 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 11 32 theme N-glycan 1557:1564 arg1 markers 1566:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 32 theme N-glycan 1557:1564 arg1 alternatives 1621:1632 novel, valuable and noninvasive alternatives 1589:1632 novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring 1589:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 32 theme N-glycan 1557:1564 arg1 models 1578:1583 models 1578:1583 models 1578:1583 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 5 33 theme fluorophore-assisted 820:839 arg1 DSA-FACE 868:875 DSA-FACE 868:875 DSA-FACE 868:875 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 33 theme fluorophore-assisted 820:839 arg1 electrophoresis 851:865 fluorophore-assisted capillary electrophoresis 820:865 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 0 34 theme Serum 0:4 arg1 N-glycans 6:14 Serum N-glycans 0:14 Serum N-glycans 0:14 Serum N-glycans outperform CA19-9 in diagnosis of extrahepatic cholangiocarcinoma. 28752594 6 35 theme carbohydrate 961:972 arg1 19-9 982:985 carbohydrate antigen 19-9 961:985 carbohydrate antigen 19-9 961:985 Multi-parameter models were constructed by combining the N-glycans and carbohydrate antigen 19-9 (CA19-9) which is currently used clinically. 28752594 7 36 theme N-glycan 1075:1082 arg1 structures 1084:1093 13 N-glycan structures 1072:1093 13 N-glycan structures 1072:1093 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 5 37 theme capillary 841:849 arg1 DSA-FACE 868:875 DSA-FACE 868:875 DSA-FACE 868:875 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 37 theme capillary 841:849 arg1 electrophoresis 851:865 fluorophore-assisted capillary electrophoresis 820:865 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 1 38 theme silent 197:202 arg1 character 213:221 its silent clinical character 193:221 its silent clinical character 193:221 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 4 39 from patients 551:558 arg1 samples 538:544 255 serum samples 528:544 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) 528:653 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 11 40 theme Serum 1551:1555 arg1 markers 1566:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 40 theme Serum 1551:1555 arg1 alternatives 1621:1632 novel, valuable and noninvasive alternatives 1589:1632 novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring 1589:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 40 theme Serum 1551:1555 arg1 models 1578:1583 models 1578:1583 models 1578:1583 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 9 41 dep NA3Fb 1380:1384 arg1 peak12 1387:1392 peak12 1387:1392 peak12 1387:1392 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 2 42 from alterations 277:287 arg1 N-glycosylation 292:306 N-glycosylation 292:306 N-glycosylation of glycoproteins 292:323 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 1 43 theme clinical 204:211 arg1 character 213:221 its silent clinical character 193:221 its silent clinical character 193:221 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 10 44 theme profiling 1531:1539 arg1 patterns 1541:1548 relatively specific N-glycome profiling patterns 1501:1548 relatively specific N-glycome profiling patterns 1501:1548 In conclusion, sera of ECCA showed relatively specific N-glycome profiling patterns. 28752594 4 45 with controls 633:640 arg1 disease 599:605 benign bile tract disease 581:605 benign bile tract disease (BBD, n = 60) 581:619 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 45 with controls 633:640 arg1 ECCA 565:568 ECCA (n = 106) 565:578 ECCA (n = 106) 565:578 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 8 46 theme single 1307:1312 arg1 N-glycan 1314:1321 single N-glycan 1307:1321 single N-glycan 1307:1321 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 7 47 gly bifucosylated 1100:1112 arg1 N-glycan 1127:1134 the bifucosylated triantennary N-glycan 1096:1134 the bifucosylated triantennary N-glycan (peak10, NA3F2) 1096:1150 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 4 48 theme healthy 625:631 arg1 controls 633:640 healthy controls 625:640 healthy controls (HC, n = 89) 625:653 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 49 theme serum 532:536 arg1 samples 538:544 255 serum samples 528:544 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) 528:653 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 5 50 theme Only 671:674 arg1 2 μL 676:679 Only 2 μL 671:679 Only 2 μL of serum from individual patients 671:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 51 from 2 μL 676:679 arg1 patients 706:713 individual patients 695:713 individual patients 695:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 9 52 theme N-glycan 1352:1359 arg1 structures 1361:1370 two N-glycan structures 1348:1370 two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) 1348:1400 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 10 53 theme N-glycome 1521:1529 arg1 patterns 1541:1548 relatively specific N-glycome profiling patterns 1501:1548 relatively specific N-glycome profiling patterns 1501:1548 In conclusion, sera of ECCA showed relatively specific N-glycome profiling patterns. 28752594 0 54 theme extrahepatic 50:61 arg1 cholangiocarcinoma 63:80 extrahepatic cholangiocarcinoma 50:80 extrahepatic cholangiocarcinoma 50:80 Serum N-glycans outperform CA19-9 in diagnosis of extrahepatic cholangiocarcinoma. 28752594 5 55 theme electrophoresis 851:865 arg1 technology 878:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 7 56 dep N-glycan 1127:1134 arg1 NA3F2 1145:1149 NA3F2 1145:1149 NA3F2 1145:1149 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 7 56 dep N-glycan 1127:1134 arg1 peak10 1137:1142 peak10 1137:1142 peak10 1137:1142 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 3 57 theme serum 485:489 arg1 markers 500:506 potential serum N-glycan markers 475:506 potential serum N-glycan markers for ECCA 475:515 This study aims to find potential serum N-glycan markers for ECCA. 28752594 11 58 theme noninvasive 1609:1619 arg1 markers 1566:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 58 theme noninvasive 1609:1619 arg1 alternatives 1621:1632 novel, valuable and noninvasive alternatives 1589:1632 novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring 1589:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 58 theme noninvasive 1609:1619 arg1 models 1578:1583 models 1578:1583 models 1578:1583 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 7 59 theme Quantitative 1032:1043 arg1 analyses 1045:1052 Quantitative analyses 1032:1052 Quantitative analyses 1032:1052 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 9 60 theme node 1432:1435 arg1 metastasis 1437:1446 lymph node metastasis 1426:1446 lymph node metastasis in ECCA patients 1426:1463 Additionally, two N-glycan structures (peak9, NA3Fb; peak12, NA4Fb) were tightly related to lymph node metastasis in ECCA patients. 28752594 2 61 theme cancer 359:364 arg1 progression 366:376 cancer progression 359:376 cancer progression 359:376 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 11 62 theme progression 1656:1666 arg1 monitoring 1668:1677 progression monitoring 1656:1677 progression monitoring 1656:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 4 63 dep disease 599:605 arg1 n = 60 613:618 n = 60 613:618 n = 60 613:618 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 63 dep disease 599:605 arg1 BBD 608:610 BBD 608:610 BBD 608:610 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 8 64 theme diagnostic 1239:1248 arg1 Glycotest2 1273:1282 Glycotest2 1273:1282 Glycotest2 1273:1282 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 8 64 theme diagnostic 1239:1248 arg1 Glycotest1 1258:1267 Glycotest1 1258:1267 Glycotest1 1258:1267 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 8 64 theme diagnostic 1239:1248 arg1 models 1250:1255 Two diagnostic models 1235:1255 Two diagnostic models (Glycotest1 and Glycotest2) 1235:1283 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 3 65 theme N-glycan 491:498 arg1 markers 500:506 potential serum N-glycan markers 475:506 potential serum N-glycan markers for ECCA 475:515 This study aims to find potential serum N-glycan markers for ECCA. 28752594 4 66 theme tract 593:597 arg1 ECCA 565:568 ECCA (n = 106) 565:578 ECCA (n = 106) 565:578 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 66 theme tract 593:597 arg1 disease 599:605 benign bile tract disease 581:605 benign bile tract disease (BBD, n = 60) 581:619 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 8 67 dep models 1250:1255 arg1 Glycotest2 1273:1282 Glycotest2 1273:1282 Glycotest2 1273:1282 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 8 67 dep models 1250:1255 arg1 Glycotest1 1258:1267 Glycotest1 1258:1267 Glycotest1 1258:1267 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 8 67 dep models 1250:1255 arg1 models 1250:1255 Two diagnostic models 1235:1255 Two diagnostic models (Glycotest1 and Glycotest2) 1235:1283 Two diagnostic models (Glycotest1 and Glycotest2) performed better than single N-glycan or CA19-9. 28752594 2 68 theme Specific 268:275 arg1 alterations 277:287 Specific alterations 268:287 Specific alterations in N-glycosylation of glycoproteins 268:323 Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. 28752594 11 69 theme novel 1589:1593 arg1 markers 1566:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers 1551:1572 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 69 theme novel 1589:1593 arg1 alternatives 1621:1632 novel, valuable and noninvasive alternatives 1589:1632 novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring 1589:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 69 theme novel 1589:1593 arg1 models 1578:1583 models 1578:1583 models 1578:1583 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 4 70 theme bile 588:591 arg1 ECCA 565:568 ECCA (n = 106) 565:578 ECCA (n = 106) 565:578 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 4 70 theme bile 588:591 arg1 disease 599:605 benign bile tract disease 581:605 benign bile tract disease (BBD, n = 60) 581:619 In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy controls (HC, n = 89) were recruited. 28752594 5 71 theme individual 695:704 arg1 patients 706:713 individual patients 695:713 individual patients 695:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 72 from patients 706:713 arg1 serum 684:688 serum 684:688 serum from individual patients 684:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 5 72 from patients 706:713 arg1 2 μL 676:679 Only 2 μL 671:679 Only 2 μL of serum from individual patients 671:713 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 3 73 theme potential 475:483 arg1 markers 500:506 potential serum N-glycan markers 475:506 potential serum N-glycan markers for ECCA 475:515 This study aims to find potential serum N-glycan markers for ECCA. 28752594 1 74 theme extrahepatic 147:158 arg1 cholangiocarcinoma 160:177 extrahepatic cholangiocarcinoma 147:177 extrahepatic cholangiocarcinoma (ECCA) 147:184 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 1 74 theme extrahepatic 147:158 arg1 ECCA 180:183 ECCA 180:183 ECCA 180:183 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 5 75 theme sequencer-assisted 801:818 arg1 technology 878:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology 797:887 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 0 76 theme cholangiocarcinoma 63:80 arg1 diagnosis 37:45 diagnosis 37:45 diagnosis of extrahepatic cholangiocarcinoma 37:80 Serum N-glycans outperform CA19-9 in diagnosis of extrahepatic cholangiocarcinoma. 28752594 7 77 theme diagnostic 1171:1180 arg1 performance 1182:1192 the best diagnostic performance 1162:1192 the best diagnostic performance for distinguishing ECCA from BBD and HC 1162:1232 Quantitative analyses showed that among 13 N-glycan structures, the bifucosylated triantennary N-glycan (peak10, NA3F2) presented the best diagnostic performance for distinguishing ECCA from BBD and HC. 28752594 5 78 theme serum 761:765 arg1 glycoproteins 767:779 serum glycoproteins 761:779 serum glycoproteins 761:779 Only 2 μL of serum from individual patients was used in this assay where the N-glycome of serum glycoproteins was profiled by DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) technology. 28752594 1 79 theme cholangiocarcinoma 160:177 arg1 diagnosis 134:142 the diagnosis 130:142 the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character 130:221 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 1 79 theme cholangiocarcinoma 160:177 arg1 lack 227:230 lack 227:230 lack of effective diagnostic biomarkers 227:265 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 28752594 11 80 from alternatives 1621:1632 arg1 diagnosis 1642:1650 ECCA diagnosis 1637:1650 ECCA diagnosis 1637:1650 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 11 80 from alternatives 1621:1632 arg1 monitoring 1668:1677 progression monitoring 1656:1677 progression monitoring 1656:1677 Serum N-glycan markers and models are novel, valuable and noninvasive alternatives in ECCA diagnosis and progression monitoring. 28752594 1 81 theme diagnostic 245:254 arg1 biomarkers 256:265 effective diagnostic biomarkers 235:265 effective diagnostic biomarkers 235:265 Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. 26051934 2 0 dep glycan 370:375 arg1 shield 377:382 shield 377:382 shield 377:382 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 3 1 from role 432:435 arg1 recognition 445:455 bNAb recognition 440:455 bNAb recognition 440:455 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 1 2 theme cell 153:156 arg1 entry 158:162 HIV-1 cell entry 147:162 HIV-1 cell entry 147:162 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 6 3 from glycans 883:889 arg1 proteins 933:940 uncleaved, non-native oligomeric gp140 proteins 894:940 uncleaved, non-native oligomeric gp140 proteins 894:940 The glycans on uncleaved, non-native oligomeric gp140 proteins are also highly processed. 26051934 5 4 contain carry 842:846 arg2 glycans 870:876 more highly processed glycans 848:876 more highly processed glycans 848:876 In contrast, gp41 subunits carry more highly processed glycans. 26051934 5 4 contain carry 842:846 arg1 subunits 833:840 gp41 subunits 828:840 gp41 subunits 828:840 In contrast, gp41 subunits carry more highly processed glycans. 26051934 5 4 contain carry 842:846 arg3 contrast 818:825 contrast 818:825 contrast 818:825 In contrast, gp41 subunits carry more highly processed glycans. 26051934 7 5 theme potential 1081:1089 arg1 heel 1101:1104 a potential Achilles' heel 1079:1104 a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design 1079:1166 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 2 6 dep density 174:180 arg1 The 165:167 The 165:167 The 165:167 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 1 7 gly glycosylated 87:98 arg1 Env 133:135 Env 133:135 Env 133:135 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 1 7 gly glycosylated 87:98 arg1 glycoprotein 119:130 A highly glycosylated, trimeric envelope glycoprotein 78:130 A highly glycosylated, trimeric envelope glycoprotein (Env) 78:136 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 7 8 theme Achilles 1091:1098 arg1 heel 1101:1104 a potential Achilles' heel 1079:1104 a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design 1079:1166 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 1 9 dep glycosylated 87:98 arg1 trimeric 101:108 trimeric 101:108 trimeric 101:108 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 7 10 dep homogeneous 971:981 arg1 oligomannose-dominated 984:1005 oligomannose-dominated 984:1005 oligomannose-dominated 984:1005 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 7 11 theme vaccine 1153:1159 arg1 design 1161:1166 vaccine design 1153:1166 vaccine design 1153:1166 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 4 12 theme trimer 710:715 arg1 components 692:701 the gp120 components 682:701 the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification 682:812 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 4 13 theme cell 748:751 arg1 system 764:769 the mammalian cell expression system 734:769 the mammalian cell expression system 734:769 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 7 14 theme native 1051:1056 arg1 conformation 1062:1073 a native Env conformation 1049:1073 a native Env conformation 1049:1073 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 3 15 dep soluble 477:483 arg1 cleaved 486:492 cleaved 486:492 cleaved 486:492 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 7 16 theme conformation 1062:1073 arg1 profile 1014:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 7 16 theme conformation 1062:1073 arg1 hallmark 1037:1044 a hallmark 1035:1044 a hallmark of a native Env conformation 1035:1073 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 7 16 theme conformation 1062:1073 arg1 heel 1101:1104 a potential Achilles' heel 1079:1104 a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design 1079:1166 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 5 17 theme gp41 828:831 arg1 subunits 833:840 gp41 subunits 828:840 gp41 subunits 828:840 In contrast, gp41 subunits carry more highly processed glycans. 26051934 7 18 theme Env 1058:1060 arg1 conformation 1062:1073 a native Env conformation 1049:1073 a native Env conformation 1049:1073 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 1 19 theme glycosylated 87:98 arg1 Env 133:135 Env 133:135 Env 133:135 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 1 19 theme glycosylated 87:98 arg1 glycoprotein 119:130 A highly glycosylated, trimeric envelope glycoprotein 78:130 A highly glycosylated, trimeric envelope glycoprotein (Env) 78:136 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 0 20 theme Structural 0:9 arg1 Constraints 11:21 Structural Constraints 0:21 Structural Constraints 0:21 Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. 26051934 6 21 theme uncleaved 894:902 arg1 proteins 933:940 uncleaved, non-native oligomeric gp140 proteins 894:940 uncleaved, non-native oligomeric gp140 proteins 894:940 The glycans on uncleaved, non-native oligomeric gp140 proteins are also highly processed. 26051934 2 22 theme potent 289:294 arg1 bNAbs 329:333 bNAbs 329:333 bNAbs 329:333 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 2 22 theme potent 289:294 arg1 antibodies 317:326 many potent broadly neutralizing antibodies 284:326 many potent broadly neutralizing antibodies (bNAbs) 284:334 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 3 23 dep mimic 572:576 arg1 a 539:539 a 539:539 a 539:539 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 4 24 dep Large 593:597 arg1 unprocessed 600:610 unprocessed 600:610 unprocessed 600:610 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 2 25 theme many 284:287 arg1 bNAbs 329:333 bNAbs 329:333 bNAbs 329:333 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 2 25 theme many 284:287 arg1 antibodies 317:326 many potent broadly neutralizing antibodies 284:326 many potent broadly neutralizing antibodies (bNAbs) 284:334 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 7 26 theme bNAb 1132:1135 arg1 recognition 1137:1147 bNAb recognition 1132:1147 bNAb recognition 1132:1147 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 1 27 gly glycoprotein 119:130 arg1 Env 133:135 Env 133:135 Env 133:135 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 1 27 gly glycoprotein 119:130 arg1 glycoprotein 119:130 A highly glycosylated, trimeric envelope glycoprotein 78:130 A highly glycosylated, trimeric envelope glycoprotein (Env) 78:136 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 1 28 theme envelope 110:117 arg1 Env 133:135 Env 133:135 Env 133:135 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 1 28 theme envelope 110:117 arg1 glycoprotein 119:130 A highly glycosylated, trimeric envelope glycoprotein 78:130 A highly glycosylated, trimeric envelope glycoprotein (Env) 78:136 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 26051934 4 29 theme Large 593:597 arg1 Man8-9GlcNAc2 642:654 Man8-9GlcNAc2 642:654 Man8-9GlcNAc2 642:654 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 4 29 theme Large 593:597 arg1 structures 630:639 Large, unprocessed oligomannose-type structures 593:639 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) 593:655 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 6 30 theme gp140 927:931 arg1 proteins 933:940 uncleaved, non-native oligomeric gp140 proteins 894:940 uncleaved, non-native oligomeric gp140 proteins 894:940 The glycans on uncleaved, non-native oligomeric gp140 proteins are also highly processed. 26051934 2 31 theme high 169:172 arg1 density 174:180 high density 169:180 high density 169:180 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 5 32 theme processed 860:868 arg1 glycans 870:876 more highly processed glycans 848:876 more highly processed glycans 848:876 In contrast, gp41 subunits carry more highly processed glycans. 26051934 4 33 theme oligomannose-type 612:628 arg1 Man8-9GlcNAc2 642:654 Man8-9GlcNAc2 642:654 Man8-9GlcNAc2 642:654 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 4 33 theme oligomannose-type 612:628 arg1 structures 630:639 Large, unprocessed oligomannose-type structures 593:639 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) 593:655 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 3 34 theme Env 406:408 arg1 glycosylation 410:422 Env glycosylation 406:422 Env glycosylation 406:422 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 6 35 theme oligomeric 916:925 arg1 proteins 933:940 uncleaved, non-native oligomeric gp140 proteins 894:940 uncleaved, non-native oligomeric gp140 proteins 894:940 The glycans on uncleaved, non-native oligomeric gp140 proteins are also highly processed. 26051934 4 36 theme bNAb 778:781 arg1 irrespective 718:729 irrespective 718:729 irrespective 718:729 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 3 37 theme bNAb 440:443 arg1 recognition 445:455 bNAb recognition 440:455 bNAb recognition 440:455 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 0 38 theme Envelope 60:67 arg1 Trimers 69:75 HIV-1 Envelope Trimers 54:75 HIV-1 Envelope Trimers 54:75 Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. 26051934 6 39 dep uncleaved 894:902 arg1 non-native 905:914 non-native 905:914 non-native 905:914 The glycans on uncleaved, non-native oligomeric gp140 proteins are also highly processed. 26051934 4 40 theme affinity 792:799 arg1 purification 801:812 affinity purification 792:812 affinity purification 792:812 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 7 41 theme glycan 1007:1012 arg1 profile 1014:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 7 41 theme glycan 1007:1012 arg1 hallmark 1037:1044 a hallmark 1035:1044 a hallmark of a native Env conformation 1035:1073 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 4 42 theme gp120 686:690 arg1 components 692:701 the gp120 components 682:701 the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification 682:812 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 0 43 theme HIV-1 54:58 arg1 Trimers 69:75 HIV-1 Envelope Trimers 54:75 HIV-1 Envelope Trimers 54:75 Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. 26051934 2 44 theme glycans 207:213 arg1 heterogeneity 186:198 heterogeneity 186:198 heterogeneity 186:198 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 2 44 theme glycans 207:213 arg1 density 174:180 high density 169:180 high density 169:180 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 7 45 theme homogeneous 971:981 arg1 profile 1014:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile 969:1020 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 7 45 theme homogeneous 971:981 arg1 hallmark 1037:1044 a hallmark 1035:1044 a hallmark of a native Env conformation 1035:1073 A homogeneous, oligomannose-dominated glycan profile is therefore a hallmark of a native Env conformation and a potential Achilles' heel that can be exploited for bNAb recognition and vaccine design. 26051934 3 46 theme soluble 477:483 arg1 SOSIP.664 520:528 BG505 SOSIP.664 514:528 BG505 SOSIP.664 514:528 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 3 46 theme soluble 477:483 arg1 mimic 572:576 mimic 572:576 mimic 572:576 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 3 46 theme soluble 477:483 arg1 trimer 506:511 a soluble, cleaved recombinant trimer 475:511 a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env 475:590 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 3 47 theme native 581:586 arg1 Env 588:590 native Env 581:590 native Env 581:590 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 0 48 gly Glycosylation 37:49 arg1 Trimers 69:75 HIV-1 Envelope Trimers 54:75 HIV-1 Envelope Trimers 54:75 Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. 26051934 2 49 theme immune 250:255 arg1 system 257:262 the immune system 246:262 the immune system 246:262 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 4 50 theme irrespective 718:729 arg1 trimer 710:715 the trimer 706:715 the trimer 706:715 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 4 51 theme mammalian 738:746 arg1 system 764:769 the mammalian cell expression system 734:769 the mammalian cell expression system 734:769 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 4 52 theme expression 753:762 arg1 system 764:769 the mammalian cell expression system 734:769 the mammalian cell expression system 734:769 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 3 53 from glycosylation 410:422 arg1 recognition 445:455 bNAb recognition 440:455 bNAb recognition 440:455 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 2 54 dep potent 289:294 arg1 neutralizing 304:315 neutralizing 304:315 neutralizing 304:315 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 4 55 theme system 764:769 arg1 irrespective 718:729 irrespective 718:729 irrespective 718:729 Large, unprocessed oligomannose-type structures (Man8-9GlcNAc2) are notably prevalent on the gp120 components of the trimer, irrespective of the mammalian cell expression system or the bNAb used for affinity purification. 26051934 3 56 theme recombinant 494:504 arg1 SOSIP.664 520:528 BG505 SOSIP.664 514:528 BG505 SOSIP.664 514:528 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 3 56 theme recombinant 494:504 arg1 mimic 572:576 mimic 572:576 mimic 572:576 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 3 56 theme recombinant 494:504 arg1 trimer 506:511 a soluble, cleaved recombinant trimer 475:511 a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env 475:590 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 2 57 gly heterogeneity 186:198 arg1 glycans 207:213 the glycans 203:213 the glycans 203:213 The high density and heterogeneity of the glycans shield Env from recognition by the immune system, but paradoxically, many potent broadly neutralizing antibodies (bNAbs) recognize epitopes involving this glycan shield. 26051934 0 58 theme Trimers 69:75 arg1 Glycosylation 37:49 the Glycosylation 33:49 the Glycosylation of HIV-1 Envelope Trimers 33:75 Structural Constraints Determine the Glycosylation of HIV-1 Envelope Trimers. 26051934 3 59 gly glycosylation 410:422 arg1 recognition 445:455 bNAb recognition 440:455 bNAb recognition 440:455 To better understand Env glycosylation and its role in bNAb recognition, we characterized a soluble, cleaved recombinant trimer (BG505 SOSIP.664) that is a close structural and antigenic mimic of native Env. 26051934 1 60 theme HIV-1 147:151 arg1 entry 158:162 HIV-1 cell entry 147:162 HIV-1 cell entry 147:162 A highly glycosylated, trimeric envelope glycoprotein (Env) mediates HIV-1 cell entry. 28890404 6 0 theme similar 1137:1143 arg1 patterns 1145:1152 similar patterns 1137:1152 similar patterns of secretion and activity 1137:1178 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 5 1 theme non-glycosylated 1001:1016 arg1 enzyme 1018:1023 the non-glycosylated enzyme 997:1023 the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) 997:1053 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 2 2 from biomass 374:380 arg1 production 352:361 biofuel production 344:361 biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants 344:471 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 1 3 theme key 181:183 arg1 Enzymes 169:175 Carbohydrate-Active Enzymes 149:175 Carbohydrate-Active Enzymes 149:175 Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. 28890404 1 3 theme key 181:183 arg1 enzymes 185:191 key enzymes 181:191 key enzymes for biomass-to-bioproducts conversion 181:229 Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. 28890404 0 4 theme Aspergillus 100:110 arg1 nidulans 112:119 Aspergillus nidulans 100:119 Aspergillus nidulans grown on sugarcane bagasse 100:146 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 10 5 theme catalytic 1867:1875 arg1 efficiency 1877:1886 catalytic efficiency 1867:1886 catalytic efficiency 1867:1886 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 9 6 theme energy 1681:1686 arg1 barriers 1688:1695 moderately low energy barriers 1666:1695 moderately low energy barriers 1666:1695 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 10 7 theme bond 1830:1833 arg1 vicinity 1793:1800 the vicinity 1789:1800 the vicinity of the arabinose glycosidic bond 1789:1833 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 3 8 theme Aspergillus 576:586 arg1 nidulans 588:595 Aspergillus nidulans 576:595 Aspergillus nidulans 576:595 In this work, we identified a GH62 α-l-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse. 28890404 10 9 theme conformations 1711:1723 arg1 One 1698:1700 One 1698:1700 One 1698:1700 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 10 9 theme conformations 1711:1723 arg1 conformations 1711:1723 these conformations 1705:1723 these conformations 1705:1723 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 2 10 theme important 318:326 arg1 applications 328:339 important applications 318:339 important applications 318:339 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 3 11 theme GH62 504:507 arg1 α-l-arabinofuranosidase 509:531 a GH62 α-l-arabinofuranosidase 502:531 a GH62 α-l-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse 502:631 In this work, we identified a GH62 α-l-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse. 28890404 3 11 theme GH62 504:507 arg1 AnAbf62Awt 534:543 AnAbf62Awt 534:543 AnAbf62Awt 534:543 In this work, we identified a GH62 α-l-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse. 28890404 4 12 gly N-glycosylated 769:782 arg1 N-glycosylated 769:782 N-glycosylated 769:782 N-glycosylated 769:782 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 4 12 gly N-glycosylated 769:782 arg1 enzyme 759:764 the enzyme 755:764 the enzyme 755:764 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 4 12 gly N-glycosylated 769:782 arg2 asparagine 787:796 asparagine 83 787:799 asparagine 83 787:799 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 4 13 from asparagine 787:796 arg1 N-glycosylated 769:782 N-glycosylated 769:782 N-glycosylated 769:782 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 4 13 from asparagine 787:796 arg1 enzyme 759:764 the enzyme 755:764 the enzyme 755:764 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 4 14 theme mass 804:807 arg1 analysis 822:829 mass spectrometry analysis 804:829 mass spectrometry analysis 804:829 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 6 15 theme designed 1093:1100 arg1 AnAbf62AN83Q 1114:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 7 16 theme five-bladed 1379:1389 arg1 fold 1403:1406 a five-bladed β-propeller fold 1377:1406 a five-bladed β-propeller fold 1377:1406 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 5 17 dep temperature 944:954 arg1 the 932:934 the 932:934 the 932:934 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 2 18 theme primary 430:436 arg1 walls 457:461 both the primary and secondary cell walls 421:461 walls 457:461 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 10 19 theme glycosidic 1819:1828 arg1 bond 1830:1833 the arabinose glycosidic bond 1805:1833 the arabinose glycosidic bond 1805:1833 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 5 20 from coli 1033:1036 arg1 AnAbf62Awt 982:991 AnAbf62Awt 982:991 AnAbf62Awt 982:991 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 20 from coli 1033:1036 arg1 temperature 944:954 melting temperature 936:954 melting temperature 936:954 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 20 from coli 1033:1036 arg1 profile 971:977 structural profile 960:977 structural profile 960:977 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 20 from coli 1033:1036 arg1 enzyme 1018:1023 the non-glycosylated enzyme 997:1023 the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) 997:1053 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 1 21 theme biomass-to-bioproducts 197:218 arg1 conversion 220:229 biomass-to-bioproducts conversion 197:229 biomass-to-bioproducts conversion 197:229 Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. 28890404 0 22 theme Structural 0:9 arg1 characterization 26:41 Structural and functional characterization 0:41 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 0:147 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 9 23 theme low 1677:1679 arg1 barriers 1688:1695 moderately low energy barriers 1666:1695 moderately low energy barriers 1666:1695 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 4 24 theme homologous 696:705 arg1 production 707:716 homologous production 696:716 homologous production of AnAbf62Awt 696:730 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 8 25 theme substrate-binding 1518:1534 arg1 residue 1536:1542 an important substrate-binding residue 1505:1542 an important substrate-binding residue 1505:1542 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 8 25 theme substrate-binding 1518:1534 arg1 Y312 1496:1499 Y312 1496:1499 Y312 1496:1499 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 2 26 contain have 313:316 arg1 α-l-Arabinofuranosidases 232:255 α-l-Arabinofuranosidases 232:255 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) 232:311 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 2 26 contain have 313:316 arg2 applications 328:339 important applications 318:339 important applications 318:339 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 0 27 theme sugarcane 130:138 arg1 bagasse 140:146 sugarcane bagasse 130:146 sugarcane bagasse 130:146 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 5 28 theme structural 960:969 arg1 profile 971:977 structural profile 960:977 structural profile 960:977 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 2 29 theme biofuel 344:350 arg1 production 352:361 biofuel production 344:361 biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants 344:471 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 7 30 theme family 1431:1436 arg1 GH62 1438:1441 family GH62 1431:1441 family GH62 1431:1441 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 7 31 theme resolution 1353:1362 arg1 model 1364:1368 the 1.7Å resolution model 1344:1368 the 1.7Å resolution model 1344:1368 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 2 32 theme plants 466:471 arg1 walls 457:461 both the primary and secondary cell walls 421:461 walls 457:461 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 5 33 theme enzyme 1018:1023 arg1 temperature 944:954 melting temperature 936:954 melting temperature 936:954 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 33 theme enzyme 1018:1023 arg1 profile 971:977 structural profile 960:977 structural profile 960:977 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 9 34 theme dynamics 1555:1562 arg1 simulations 1564:1574 Molecular dynamics simulations 1545:1574 Molecular dynamics simulations 1545:1574 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 7 35 theme crystallographic 1282:1297 arg1 structure 1299:1307 The crystallographic structure 1278:1307 The crystallographic structure of AnAbf62Adeglyc 1278:1325 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 4 36 theme AnAbf62Awt 721:730 arg1 production 707:716 homologous production 696:716 homologous production of AnAbf62Awt 696:730 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 9 37 theme Molecular 1545:1553 arg1 simulations 1564:1574 Molecular dynamics simulations 1545:1574 Molecular dynamics simulations 1545:1574 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 2 38 theme Hydrolase 286:294 arg1 family 296:301 the Glycoside Hydrolase family 62 272:304 the Glycoside Hydrolase family 62 (GH62) 272:311 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 2 38 theme Hydrolase 286:294 arg1 GH62 307:310 GH62 307:310 GH62 307:310 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 5 39 theme circular 901:908 arg1 dichroism 910:918 circular dichroism 901:918 circular dichroism 901:918 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 2 40 theme cell 452:455 arg1 walls 457:461 both the primary and secondary cell walls 421:461 walls 457:461 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 6 41 attach presented 1127:1135 arg2 AnAbf62AN83Q 1114:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 6 41 attach presented 1127:1135 arg1 addition 1079:1086 addition 1079:1086 addition 1079:1086 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 2 42 theme Glycoside 276:284 arg1 family 296:301 the Glycoside Hydrolase family 62 272:304 the Glycoside Hydrolase family 62 (GH62) 272:311 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 2 42 theme Glycoside 276:284 arg1 GH62 307:310 GH62 307:310 GH62 307:310 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 2 43 theme secondary 442:450 arg1 walls 457:461 both the primary and secondary cell walls 421:461 walls 457:461 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 0 44 theme secreted 55:62 arg1 GH62 89:92 GH62 89:92 GH62 89:92 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 0 44 theme secreted 55:62 arg1 α-l-arabinofuranosidase 64:86 a highly secreted α-l-arabinofuranosidase 46:86 a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse 46:146 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 5 45 theme dichroism 910:918 arg1 studies 890:896 the studies 886:896 the studies of circular dichroism 886:918 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 8 46 theme important 1508:1516 arg1 residue 1536:1542 an important substrate-binding residue 1505:1542 an important substrate-binding residue 1505:1542 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 8 46 theme important 1508:1516 arg1 Y312 1496:1499 Y312 1496:1499 Y312 1496:1499 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 3 47 theme sugarcane 615:623 arg1 bagasse 625:631 sugarcane bagasse 615:631 sugarcane bagasse 615:631 In this work, we identified a GH62 α-l-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse. 28890404 5 48 theme melting 936:942 arg1 temperature 944:954 melting temperature 936:954 melting temperature 936:954 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 10 49 theme arabinose 1809:1817 arg1 bond 1830:1833 the arabinose glycosidic bond 1805:1833 the arabinose glycosidic bond 1805:1833 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 0 50 from characterization 26:41 arg1 nidulans 112:119 Aspergillus nidulans 100:119 Aspergillus nidulans grown on sugarcane bagasse 100:146 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 2 51 dep walls 457:461 arg1 both 421:424 both 421:424 both 421:424 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 7 52 theme β-propeller 1391:1401 arg1 fold 1403:1406 a five-bladed β-propeller fold 1377:1406 a five-bladed β-propeller fold 1377:1406 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 6 53 theme activity 1171:1178 arg1 patterns 1145:1152 similar patterns 1137:1152 similar patterns of secretion and activity 1137:1178 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 5 54 from temperature 944:954 arg1 AnAbf62Adeglyc 1039:1052 AnAbf62Adeglyc 1039:1052 AnAbf62Adeglyc 1039:1052 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 54 from temperature 944:954 arg1 coli 1033:1036 E. coli 1030:1036 E. coli (AnAbf62Adeglyc) 1030:1053 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 0 55 from nidulans 112:119 arg1 GH62 89:92 GH62 89:92 GH62 89:92 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 0 55 from nidulans 112:119 arg1 α-l-arabinofuranosidase 64:86 a highly secreted α-l-arabinofuranosidase 46:86 a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse 46:146 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 0 55 from nidulans 112:119 arg1 characterization 26:41 Structural and functional characterization 0:41 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 0:147 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 6 56 theme enzyme 1270:1275 arg1 properties 1251:1260 the properties 1247:1260 the properties of this enzyme 1247:1275 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 7 57 theme AnAbf62Adeglyc 1312:1325 arg1 structure 1299:1307 The crystallographic structure 1278:1307 The crystallographic structure of AnAbf62Adeglyc 1278:1325 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 5 58 theme AnAbf62Awt 982:991 arg1 temperature 944:954 melting temperature 936:954 melting temperature 936:954 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 58 theme AnAbf62Awt 982:991 arg1 profile 971:977 structural profile 960:977 structural profile 960:977 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 6 59 theme secretion 1157:1165 arg1 patterns 1145:1152 similar patterns 1137:1152 similar patterns of secretion and activity 1137:1178 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 8 60 dep Mutants 1444:1450 arg1 AnAbf62AY312F 1452:1464 AnAbf62AY312F 1452:1464 AnAbf62AY312F 1452:1464 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 8 60 dep Mutants 1444:1450 arg1 Mutants 1444:1450 Mutants AnAbf62AY312F and AnAbf62AY312S 1444:1482 Mutants AnAbf62AY312F and AnAbf62AY312S 1444:1482 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 8 60 dep Mutants 1444:1450 arg1 AnAbf62AY312S 1470:1482 AnAbf62AY312S 1470:1482 AnAbf62AY312S 1470:1482 Mutants AnAbf62AY312F and AnAbf62AY312S showed that Y312 was an important substrate-binding residue. 28890404 4 61 theme spectrometry 809:820 arg1 analysis 822:829 mass spectrometry analysis 804:829 mass spectrometry analysis 804:829 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 2 62 theme plant 368:372 arg1 biomass 374:380 plant biomass 368:380 plant biomass 368:380 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 5 63 from profile 971:977 arg1 AnAbf62Adeglyc 1039:1052 AnAbf62Adeglyc 1039:1052 AnAbf62Adeglyc 1039:1052 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 5 63 from profile 971:977 arg1 coli 1033:1036 E. coli 1030:1036 E. coli (AnAbf62Adeglyc) 1030:1053 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 4 64 theme gene 638:641 arg1 AN7908 643:648 The gene AN7908 634:648 The gene AN7908 634:648 The gene AN7908 was cloned and transformed in A. nidulans for homologous production of AnAbf62Awt, and we confirmed that the enzyme is N-glycosylated at asparagine 83 by mass spectrometry analysis. 28890404 5 65 gly non-glycosylated 1001:1016 arg1 enzyme 1018:1023 the non-glycosylated enzyme 997:1023 the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) 997:1053 The enzyme was also expressed in Escherichia coli and the studies of circular dichroism showed that the melting temperature and structural profile of AnAbf62Awt and the non-glycosylated enzyme from E. coli (AnAbf62Adeglyc) were highly similar. 28890404 7 66 theme 1.7Å 1348:1351 arg1 model 1364:1368 the 1.7Å resolution model 1344:1368 the 1.7Å resolution model 1344:1368 The crystallographic structure of AnAbf62Adeglyc was obtained and the 1.7Å resolution model showed a five-bladed β-propeller fold, which is conserved in family GH62. 28890404 10 67 theme local 1739:1743 arg1 minimum 1745:1751 a local minimum 1737:1751 a local minimum 1737:1751 One of these conformations, comprising a local minimum, is responsible for placing Y312 in the vicinity of the arabinose glycosidic bond, and thus, may be important for catalytic efficiency. 28890404 9 68 theme loop 1595:1598 arg1 Y312 1611:1614 the loop containing Y312 1591:1614 the loop containing Y312 1591:1614 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 1 69 theme Carbohydrate-Active 149:167 arg1 Enzymes 169:175 Carbohydrate-Active Enzymes 149:175 Carbohydrate-Active Enzymes 149:175 Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. 28890404 1 69 theme Carbohydrate-Active 149:167 arg1 enzymes 185:191 key enzymes 181:191 key enzymes for biomass-to-bioproducts conversion 181:229 Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. 28890404 6 70 theme glycomutant 1102:1112 arg1 AnAbf62AN83Q 1114:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 the designed glycomutant AnAbf62AN83Q 1089:1125 In addition, the designed glycomutant AnAbf62AN83Q presented similar patterns of secretion and activity to the AnAbf62Awt, indicating that the N-glycan does not influence the properties of this enzyme. 28890404 9 71 theme different 1629:1637 arg1 conformations 1639:1651 different conformations 1629:1651 different conformations separated by moderately low energy barriers 1629:1695 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 28890404 0 72 theme α-l-arabinofuranosidase 64:86 arg1 characterization 26:41 Structural and functional characterization 0:41 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 0:147 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 2 73 located found 412:416 arg1 walls 457:461 both the primary and secondary cell walls 421:461 walls 457:461 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 2 73 located found 412:416 arg2 arabinoxylans 397:409 arabinoxylans 397:409 arabinoxylans 397:409 α-l-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. 28890404 0 74 theme functional 15:24 arg1 characterization 26:41 Structural and functional characterization 0:41 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 0:147 Structural and functional characterization of a highly secreted α-l-arabinofuranosidase (GH62) from Aspergillus nidulans grown on sugarcane bagasse. 28890404 9 75 theme containing 1600:1609 arg1 Y312 1611:1614 the loop containing Y312 1591:1614 the loop containing Y312 1591:1614 Molecular dynamics simulations indicated that the loop containing Y312 could access different conformations separated by moderately low energy barriers. 24668806 0 0 theme each 83:86 arg1 each 83:86 each 83:86 each 83:86 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 0 0 theme each 83:86 arg1 subdomains 69:78 subdomains 69:78 subdomains of each that contribute to deglycosylation 69:121 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 7 1 theme carbohydrate 922:933 arg1 CBM 951:953 CBM 951:953 CBM 951:953 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 7 1 theme carbohydrate 922:933 arg1 module 943:948 a putative carbohydrate binding module 911:948 a putative carbohydrate binding module (CBM) followed by a C-terminal region 911:986 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 5 2 theme Fc 648:649 arg1 domain 651:656 the Fc domain 644:656 the Fc domain 644:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 0 3 theme bacterial 13:21 arg1 s 39:39 bacterial endoglycosidase s 13:39 bacterial endoglycosidase s 13:39 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 10 4 theme IgG 1465:1467 arg1 glycans 1469:1475 IgG glycans 1465:1475 IgG glycans 1465:1475 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 5 5 theme domain 651:656 arg1 domain 651:656 the Fc domain 644:656 the Fc domain 644:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 5 5 theme domain 651:656 arg1 domain 634:639 the isolated CH2 domain 617:639 the isolated CH2 domain of the Fc domain 617:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 7 6 theme repeat 886:891 arg1 predictions 835:845 the previous predictions 822:845 the previous predictions of a chitinase domain and leucine-rich repeat 822:891 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 2 7 link N-linked 261:268 arg1 glycans 270:276 the N-linked glycans 257:276 the N-linked glycans from the Fc region of IgG 257:302 EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. 24668806 8 8 theme helical 1151:1157 arg1 structure 1159:1167 β sheet and α helical structure 1137:1167 structure 1159:1167 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 7 9 theme C-terminal 970:979 arg1 region 981:986 a C-terminal region 968:986 a C-terminal region 968:986 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 2 10 theme IgG 300:302 arg1 region 290:295 the Fc region 283:295 the Fc region of IgG 283:302 EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. 24668806 8 11 theme α 1149:1149 arg1 structure 1159:1167 β sheet and α helical structure 1137:1167 structure 1159:1167 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 4 12 theme deactivation 480:491 arg1 mechanism 451:459 the mechanism 447:459 the mechanism of human serum IgG deactivation by EndoS 447:500 Here, we investigate the mechanism of human serum IgG deactivation by EndoS. 24668806 8 13 theme expressed 995:1003 arg1 fragments 1005:1013 expressed fragments 995:1013 expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion 995:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 9 14 with CBM 1227:1229 arg1 monosaccharides 1236:1250 monosaccharides 1236:1250 monosaccharides 1236:1250 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 4 15 theme IgG 476:478 arg1 deactivation 480:491 human serum IgG deactivation 464:491 human serum IgG deactivation 464:491 Here, we investigate the mechanism of human serum IgG deactivation by EndoS. 24668806 5 16 theme CH2 630:632 arg1 domain 651:656 the Fc domain 644:656 the Fc domain 644:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 5 16 theme CH2 630:632 arg1 domain 634:639 the isolated CH2 domain 617:639 the isolated CH2 domain of the Fc domain 617:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 5 17 theme isolated 621:628 arg1 domain 651:656 the Fc domain 644:656 the Fc domain 644:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 5 17 theme isolated 621:628 arg1 domain 634:639 the isolated CH2 domain 617:639 the isolated CH2 domain of the Fc domain 617:656 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 7 18 theme leucine-rich 873:884 arg1 repeat 886:891 leucine-rich repeat 873:891 leucine-rich repeat 873:891 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 0 19 theme immunoglobulin 45:58 arg1 g 60:60 immunoglobulin g 45:60 immunoglobulin g 45:60 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 8 20 theme isolated 1051:1058 arg1 CBM 1060:1062 the isolated CBM 1047:1062 the isolated CBM 1047:1062 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 9 21 theme resonance 1201:1209 arg1 analysis 1211:1218 Nuclear magnetic resonance analysis 1184:1218 Nuclear magnetic resonance analysis of the CBM with monosaccharides 1184:1250 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 2 22 theme Fc 287:288 arg1 region 290:295 the Fc region 283:295 the Fc region of IgG 283:302 EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. 24668806 7 23 theme chitinase 852:860 arg1 domain 862:867 a chitinase domain 850:867 a chitinase domain 850:867 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 8 24 theme β 1137:1137 arg1 sheet 1139:1143 β sheet and α helical structure 1137:1167 sheet 1139:1143 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 7 25 theme domain 862:867 arg1 predictions 835:845 the previous predictions 822:845 the previous predictions of a chitinase domain and leucine-rich repeat 822:891 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 10 26 theme glycans 1469:1475 arg1 hydrolysis 1451:1460 the hydrolysis 1447:1460 the hydrolysis of IgG glycans 1447:1475 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 8 27 theme CBM 1060:1062 arg1 fusion 1093:1098 a CBM-C-terminal region fusion 1069:1098 a CBM-C-terminal region fusion 1069:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 8 27 theme CBM 1060:1062 arg1 EndoS 1018:1022 EndoS 1018:1022 EndoS 1018:1022 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 8 27 theme CBM 1060:1062 arg1 dichroism 1034:1042 circular dichroism 1025:1042 circular dichroism of the isolated CBM 1025:1062 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 10 28 theme EndoS 1344:1348 arg1 truncations 1329:1339 truncations 1329:1339 truncations of EndoS 1329:1348 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 8 29 theme circular 1025:1032 arg1 dichroism 1034:1042 circular dichroism 1025:1042 circular dichroism of the isolated CBM 1025:1062 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 3 30 theme immune 386:391 arg1 strategy 401:408 the immune evasion strategy 382:408 the immune evasion strategy of S. pyogenes 382:423 This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. 24668806 10 31 theme Functional 1306:1315 arg1 analysis 1317:1324 Functional analysis 1306:1324 Functional analysis of truncations of EndoS 1306:1348 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 8 32 theme CBM-C-terminal 1071:1084 arg1 fusion 1093:1098 a CBM-C-terminal region fusion 1069:1098 a CBM-C-terminal region fusion 1069:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 8 33 theme EndoS 1018:1022 arg1 fragments 1005:1013 expressed fragments 995:1013 expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion 995:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 10 34 theme truncations 1329:1339 arg1 analysis 1317:1324 Functional analysis 1306:1324 Functional analysis of truncations of EndoS 1306:1348 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 1 35 theme bacterium 191:199 arg1 pyogenes 215:222 the bacterium Streptococcus pyogenes 187:222 the bacterium Streptococcus pyogenes 187:222 Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. 24668806 3 36 theme evasion 393:399 arg1 strategy 401:408 the immune evasion strategy 382:408 the immune evasion strategy of S. pyogenes 382:423 This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. 24668806 9 37 theme CBM 1227:1229 arg1 analysis 1211:1218 Nuclear magnetic resonance analysis 1184:1218 Nuclear magnetic resonance analysis of the CBM with monosaccharides 1184:1250 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 9 38 theme Nuclear 1184:1190 arg1 resonance 1201:1209 Nuclear magnetic resonance 1184:1209 Nuclear magnetic resonance analysis of the CBM with monosaccharides 1184:1250 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 7 39 theme previous 826:833 arg1 predictions 835:845 the previous predictions 822:845 the previous predictions of a chitinase domain and leucine-rich repeat 822:891 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 1 40 theme Streptococcus 201:213 arg1 pyogenes 215:222 the bacterium Streptococcus pyogenes 187:222 the bacterium Streptococcus pyogenes 187:222 Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. 24668806 7 41 theme EndoS 806:810 arg1 organization 790:801 the domain organization 779:801 the domain organization of EndoS 779:810 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 8 42 theme fusion 1093:1098 arg1 fragments 1005:1013 expressed fragments 995:1013 expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion 995:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 0 43 theme s 39:39 arg1 Fragments 0:8 Fragments 0:8 Fragments of bacterial endoglycosidase s and immunoglobulin g 0:60 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 7 44 theme domain 783:788 arg1 organization 790:801 the domain organization 779:801 the domain organization of EndoS 779:810 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 8 45 theme region 1086:1091 arg1 fusion 1093:1098 a CBM-C-terminal region fusion 1069:1098 a CBM-C-terminal region fusion 1069:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 4 46 theme serum 470:474 arg1 deactivation 480:491 human serum IgG deactivation 464:491 human serum IgG deactivation 464:491 Here, we investigate the mechanism of human serum IgG deactivation by EndoS. 24668806 0 47 theme endoglycosidase 23:37 arg1 s 39:39 bacterial endoglycosidase s 13:39 bacterial endoglycosidase s 13:39 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 0 48 theme g 60:60 arg1 Fragments 0:8 Fragments 0:8 Fragments of bacterial endoglycosidase s and immunoglobulin g 0:60 Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation. 24668806 7 49 theme organization 790:801 arg1 analysis 767:774 Bioinformatics analysis 752:774 Bioinformatics analysis of the domain organization of EndoS 752:810 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 8 50 theme folded 1109:1114 arg1 domains 1116:1122 folded domains 1109:1122 folded domains dominated by β sheet and α helical structure 1109:1167 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 4 51 theme human 464:468 arg1 deactivation 480:491 human serum IgG deactivation 464:491 human serum IgG deactivation 464:491 Here, we investigate the mechanism of human serum IgG deactivation by EndoS. 24668806 3 52 theme pyogenes 416:423 arg1 strategy 401:408 the immune evasion strategy 382:408 the immune evasion strategy of S. pyogenes 382:423 This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. 24668806 8 53 theme dichroism 1034:1042 arg1 fragments 1005:1013 expressed fragments 995:1013 expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion 995:1098 Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. 24668806 1 54 theme Endoglycosidase 124:138 arg1 S 140:140 Endoglycosidase S 124:140 Endoglycosidase S (EndoS) 124:148 Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. 24668806 1 54 theme Endoglycosidase 124:138 arg1 glycoside-hydrolase 155:173 a glycoside-hydrolase 153:173 a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes 153:222 Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. 24668806 1 54 theme Endoglycosidase 124:138 arg1 EndoS 143:147 EndoS 143:147 EndoS 143:147 Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. 24668806 2 55 from region 290:295 arg1 glycans 270:276 the N-linked glycans 257:276 the N-linked glycans from the Fc region of IgG 257:302 EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. 24668806 7 56 theme binding 935:941 arg1 CBM 951:953 CBM 951:953 CBM 951:953 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 7 56 theme binding 935:941 arg1 module 943:948 a putative carbohydrate binding module 911:948 a putative carbohydrate binding module (CBM) followed by a C-terminal region 911:986 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 7 57 theme Bioinformatics 752:765 arg1 analysis 767:774 Bioinformatics analysis 752:774 Bioinformatics analysis of the domain organization of EndoS 752:810 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 9 58 theme carbohydrate 1270:1281 arg1 functionality 1291:1303 carbohydrate binding functionality 1270:1303 carbohydrate binding functionality 1270:1303 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 10 59 theme EndoS 1391:1395 arg1 C-terminal 1377:1386 C-terminal 1377:1386 C-terminal 1377:1386 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 10 59 theme EndoS 1391:1395 arg1 dispensable 1400:1410 dispensable 1400:1410 dispensable 1400:1410 Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans. 24668806 9 60 theme magnetic 1192:1199 arg1 resonance 1201:1209 Nuclear magnetic resonance 1184:1209 Nuclear magnetic resonance analysis of the CBM with monosaccharides 1184:1250 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 3 61 theme Fc 346:347 arg1 functionality 349:361 Fc functionality 346:361 Fc functionality 346:361 This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. 24668806 9 62 theme binding 1283:1289 arg1 functionality 1291:1303 carbohydrate binding functionality 1270:1303 carbohydrate binding functionality 1270:1303 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 5 63 theme IgG1 529:532 arg1 fragments 516:524 fragments 516:524 fragments of IgG1 516:532 We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. 24668806 7 64 theme putative 913:920 arg1 CBM 951:953 CBM 951:953 CBM 951:953 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 7 64 theme putative 913:920 arg1 module 943:948 a putative carbohydrate binding module 911:948 a putative carbohydrate binding module (CBM) followed by a C-terminal region 911:986 Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. 24668806 3 65 theme S. 413:414 arg1 pyogenes 416:423 S. pyogenes 413:423 S. pyogenes 413:423 This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. 24668806 9 66 theme functionality 1291:1303 arg1 suggestive 1256:1265 suggestive 1256:1265 suggestive 1256:1265 Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. 24668806 2 67 theme N-linked 261:268 arg1 glycans 270:276 the N-linked glycans 257:276 the N-linked glycans from the Fc region of IgG 257:302 EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. 24797265 14 0 from located 2287:2293 arg1 region 2313:2318 the N-terminal region 2298:2318 the N-terminal region of the protein 2298:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 2 1 theme Srr 366:368 arg1 proteins 370:377 Srr proteins 366:377 Srr proteins 366:377 Lectin-binding experiments have previously shown that Srr proteins are heavily glycosylated. 24797265 7 2 theme source 924:929 arg1 SpectrumFinder 946:959 SpectrumFinder 946:959 SpectrumFinder 946:959 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 7 2 theme source 924:929 arg1 tool 940:943 a new open source software tool 913:943 a new open source software tool (SpectrumFinder) developed for this work 913:984 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 8 3 gly glycopeptides 1021:1033 arg2 glycopeptides 1021:1033 27 glycopeptides 1018:1033 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] 1018:1115 By combining these approaches, 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] were identified. 24797265 7 4 theme new 915:917 arg1 SpectrumFinder 946:959 SpectrumFinder 946:959 SpectrumFinder 946:959 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 7 4 theme new 915:917 arg1 tool 940:943 a new open source software tool 913:943 a new open source software tool (SpectrumFinder) developed for this work 913:984 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 6 5 gly glycopeptides 868:880 arg2 glycopeptides 868:880 glycopeptides 868:880 glycopeptides 868:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 6 6 theme backbone 841:848 arg1 fragmentation 850:862 backbone fragmentation 841:862 backbone fragmentation for glycopeptides 841:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 2 7 theme Lectin-binding 312:325 arg1 experiments 327:337 Lectin-binding experiments 312:337 Lectin-binding experiments 312:337 Lectin-binding experiments have previously shown that Srr proteins are heavily glycosylated. 24797265 13 8 theme initial 1937:1943 arg1 step 1945:1948 an initial step 1934:1948 an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex 1934:2029 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 13 8 theme initial 1937:1943 arg1 O-GlcNAcylation 1915:1929 O-GlcNAcylation 1915:1929 O-GlcNAcylation 1915:1929 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 1 9 theme bacteria 199:206 arg1 surface 174:180 the surface 170:180 the surface of Gram-positive bacteria 170:206 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 14 10 gly glycosylation 2267:2279 arg2 sites 2281:2285 six glycosylation sites 2263:2285 six glycosylation sites located in the N-terminal region of the protein 2263:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 10 gly glycosylation 2267:2279 arg2 six 2263:2265 six 2263:2265 six 2263:2265 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 11 11 theme structural 1606:1615 arg1 diversity 1617:1625 a wide structural diversity 1599:1625 a wide structural diversity of Srr1 1599:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 13 12 from step 1945:1948 arg1 glycosylation 1958:1970 Srr1 glycosylation 1953:1970 Srr1 glycosylation 1953:1970 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 14 13 from region 2313:2318 arg1 located 2287:2293 located 2287:2293 located 2287:2293 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 6 14 theme collisional 706:716 arg1 fragmentation 731:743 higher energy collisional dissociation fragmentation 692:743 higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides 692:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 15 gly heterogeneity 2186:2198 arg1 Srr1 2203:2206 Srr1 2203:2206 Srr1 2203:2206 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 11 16 theme wide 1601:1604 arg1 diversity 1617:1625 a wide structural diversity 1599:1625 a wide structural diversity of Srr1 1599:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 0 17 theme adhesin 60:66 arg1 Srr1 68:71 the serine-rich adhesin Srr1 44:71 the serine-rich adhesin Srr1 of Streptococcus agalactiae 44:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 12 18 theme glycosylation 1686:1698 arg1 sites 1700:1704 glycosylation sites 1686:1704 glycosylation sites leading to the unambiguous identification of six serines and one threonine residues 1686:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 14 19 theme N-terminal 2302:2311 arg1 region 2313:2318 the N-terminal region 2298:2318 the N-terminal region of the protein 2298:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 8 20 theme region 1093:1098 arg1 segments 1066:1073 six different segments 1052:1073 six different segments of the N-terminal region of Srr1 [93-639 1052:1114 By combining these approaches, 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] were identified. 24797265 12 21 theme transfer 1645:1652 arg1 dissociation 1654:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 13 22 theme accessory 1852:1860 arg1 genes 1891:1895 accessory glycosyltransferase encoding genes 1852:1895 accessory glycosyltransferase encoding genes 1852:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 11 23 theme Srr1 1630:1633 arg1 diversity 1617:1625 a wide structural diversity 1599:1625 a wide structural diversity of Srr1 1599:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 0 24 theme agalactiae 90:99 arg1 Srr1 68:71 the serine-rich adhesin Srr1 44:71 the serine-rich adhesin Srr1 of Streptococcus agalactiae 44:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 8 25 theme Srr1 1103:1106 arg1 region 1093:1098 the N-terminal region 1078:1098 the N-terminal region of Srr1 [93-639 1078:1114 By combining these approaches, 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] were identified. 24797265 12 26 used used 1671:1674 arg2 dissociation 1654:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 13 27 theme Srr1 1812:1815 arg1 Analysis 1791:1798 Analysis 1791:1798 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes 1791:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 10 28 theme monosaccharide 1385:1398 arg1 analysis 1412:1419 monosaccharide composition analysis 1385:1419 monosaccharide composition analysis 1385:1419 Lectin binding and monosaccharide composition analysis strongly suggested that HexNAc and Hex correspond to N-acetylglucosamine and glucose, respectively. 24797265 14 29 theme glycosylation 2172:2184 arg1 heterogeneity 2186:2198 glycosylation heterogeneity 2172:2198 glycosylation heterogeneity of Srr1 2172:2206 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 11 30 theme same 1525:1528 arg1 segment 1538:1544 The same protein segment 1521:1544 The same protein segment 1521:1544 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 4 31 theme Srr1 518:521 arg1 enrichment 523:532 Srr1 enrichment 518:532 Srr1 enrichment 518:532 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 2 32 gly glycosylated 391:402 arg1 proteins 370:377 Srr proteins 366:377 Srr proteins 366:377 Lectin-binding experiments have previously shown that Srr proteins are heavily glycosylated. 24797265 12 33 theme one 1767:1769 arg1 residues 1781:1788 one threonine residues 1767:1788 one threonine residues 1767:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 13 34 theme subsequent 2000:2009 arg1 decoration 2011:2020 subsequent decoration 2000:2020 subsequent decoration with Hex 2000:2029 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 11 35 mod modified 1553:1560 arg3 variety 1569:1575 a variety 1567:1575 a variety of glycans generating a wide structural diversity of Srr1 1567:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 11 35 mod modified 1553:1560 arg1 segment 1538:1544 The same protein segment 1521:1544 The same protein segment 1521:1544 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 11 35 mod modified 1553:1560 arg3 glycans 1580:1586 glycans 1580:1586 glycans generating a wide structural diversity of Srr1 1580:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 1 36 theme staphylococci 280:292 arg1 virulence 256:264 the virulence 252:264 the virulence of pathogenic staphylococci and streptococci 252:309 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 14 37 theme software 2145:2152 arg1 tool 2154:2157 a new software tool 2139:2157 a new software tool 2139:2157 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 12 38 theme residues 1781:1788 arg1 identification 1733:1746 the unambiguous identification 1717:1746 the unambiguous identification of six serines and one threonine residues 1717:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 3 39 theme first 424:428 arg1 analysis 448:455 the first mass-spectrometry analysis 420:455 the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1 420:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 4 40 theme induced 610:616 arg1 spectra 631:637 collision induced dissociation spectra 600:637 collision induced dissociation spectra using X! 600:646 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 9 41 mod modified 1203:1210 arg3 modification 1297:1308 a novel modification 1289:1308 a novel modification that was identified as O-acetylated-N-acetylhexosamine 1289:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 41 mod modified 1203:1210 arg3 combinations 1234:1245 different glycan combinations 1217:1245 different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine 1217:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 41 mod modified 1203:1210 arg1 residue 1188:1194 the same protein residue 1171:1194 the same protein residue 1171:1194 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 41 mod modified 1203:1210 arg3 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 41 mod modified 1203:1210 arg3 hexose 1277:1282 hexose 1277:1282 hexose 1277:1282 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 42 theme protein 1180:1186 arg1 residue 1188:1194 the same protein residue 1171:1194 the same protein residue 1171:1194 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 0 43 theme region 34:39 arg1 O-Glycosylation 0:14 O-Glycosylation 0:14 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae 0:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 1 44 theme streptococci 298:309 arg1 virulence 256:264 the virulence 252:264 the virulence of pathogenic staphylococci and streptococci 252:309 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 12 45 theme Electron 1636:1643 arg1 dissociation 1654:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation 1636:1665 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 11 46 theme protein 1530:1536 arg1 segment 1538:1544 The same protein segment 1521:1544 The same protein segment 1521:1544 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 4 47 theme potential 557:565 arg1 glycopeptides 567:579 potential glycopeptides 557:579 potential glycopeptides 557:579 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 14 48 theme new 2224:2226 arg1 modification 2236:2247 a new protein modification 2222:2247 a new protein modification 2222:2247 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 6 49 theme oxonium 824:830 arg1 ions 832:835 diagnostic oxonium ions 813:835 diagnostic oxonium ions 813:835 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 50 theme protein 2228:2234 arg1 modification 2236:2247 a new protein modification 2222:2247 a new protein modification 2222:2247 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 51 theme new 2141:2143 arg1 tool 2154:2157 a new software tool 2139:2157 a new software tool 2139:2157 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 3 52 gly glycosylation 464:476 arg1 Srr1 506:509 Streptococcus agalactiae Srr1 481:509 Streptococcus agalactiae Srr1 481:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 3 53 dep Streptococcus 481:493 arg1 agalactiae 495:504 agalactiae 495:504 agalactiae 495:504 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 9 54 theme different 1217:1225 arg1 modification 1297:1308 a novel modification 1289:1308 a novel modification that was identified as O-acetylated-N-acetylhexosamine 1289:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 54 theme different 1217:1225 arg1 hexose 1277:1282 hexose 1277:1282 hexose 1277:1282 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 54 theme different 1217:1225 arg1 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 54 theme different 1217:1225 arg1 combinations 1234:1245 different glycan combinations 1217:1245 different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine 1217:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 1 55 theme adhesins 224:231 arg1 family 214:219 a family 212:219 a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci 212:309 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 1 55 theme adhesins 224:231 arg1 proteins 150:157 Serine-rich (Srr) proteins 132:157 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria 132:206 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 14 56 theme fragmentation 2082:2094 arg1 techniques 2114:2123 fragmentation mass spectrometry techniques 2082:2123 fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein 2082:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 6 57 theme sugar 783:787 arg1 residues 789:796 sugar residues 783:796 sugar residues 783:796 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 58 dep tool 2154:2157 arg1 identify 2254:2261 identify 2254:2261 identify six glycosylation sites located in the N-terminal region of the protein 2254:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 58 dep tool 2154:2157 arg1 demonstrate 2160:2170 demonstrate 2160:2170 demonstrate glycosylation heterogeneity of Srr1 2160:2206 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 58 dep tool 2154:2157 arg1 characterize 2209:2220 characterize 2209:2220 characterize a new protein modification 2209:2247 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 8 59 theme different 1056:1064 arg1 segments 1066:1073 six different segments 1052:1073 six different segments of the N-terminal region of Srr1 [93-639 1052:1114 By combining these approaches, 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] were identified. 24797265 4 60 theme X 645:645 arg1 ! 646:646 X! 645:646 X! 645:646 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 13 61 theme Srr1 1953:1956 arg1 glycosylation 1958:1970 Srr1 glycosylation 1953:1970 Srr1 glycosylation 1953:1970 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 7 62 theme software 931:938 arg1 SpectrumFinder 946:959 SpectrumFinder 946:959 SpectrumFinder 946:959 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 7 62 theme software 931:938 arg1 tool 940:943 a new open source software tool 913:943 a new open source software tool (SpectrumFinder) developed for this work 913:984 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 7 63 theme open 919:922 arg1 SpectrumFinder 946:959 SpectrumFinder 946:959 SpectrumFinder 946:959 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 7 63 theme open 919:922 arg1 tool 940:943 a new open source software tool 913:943 a new open source software tool (SpectrumFinder) developed for this work 913:984 This feature was exploited in a new open source software tool (SpectrumFinder) developed for this work. 24797265 8 64 theme N-terminal 1082:1091 arg1 region 1093:1098 the N-terminal region 1078:1098 the N-terminal region of Srr1 [93-639 1078:1114 By combining these approaches, 27 glycopeptides corresponding to six different segments of the N-terminal region of Srr1 [93-639] were identified. 24797265 9 65 theme novel 1291:1295 arg1 O-acetylated-N-acetylhexosamine 1333:1363 O-acetylated-N-acetylhexosamine 1333:1363 O-acetylated-N-acetylhexosamine 1333:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 65 theme novel 1291:1295 arg1 modification 1297:1308 a novel modification 1289:1308 a novel modification that was identified as O-acetylated-N-acetylhexosamine 1289:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 1 66 theme Gram-positive 185:197 arg1 bacteria 199:206 Gram-positive bacteria 185:206 Gram-positive bacteria 185:206 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 12 67 theme serines 1755:1761 arg1 identification 1733:1746 the unambiguous identification 1717:1746 the unambiguous identification of six serines and one threonine residues 1717:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 13 68 with decoration 2011:2020 arg1 Hex 2027:2029 Hex 2027:2029 Hex 2027:2029 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 13 69 theme encoding 1882:1889 arg1 genes 1891:1895 accessory glycosyltransferase encoding genes 1852:1895 accessory glycosyltransferase encoding genes 1852:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 12 70 gly glycosylation 1686:1698 arg2 sites 1700:1704 glycosylation sites 1686:1704 glycosylation sites leading to the unambiguous identification of six serines and one threonine residues 1686:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 12 71 theme unambiguous 1721:1731 arg1 identification 1733:1746 the unambiguous identification 1717:1746 the unambiguous identification of six serines and one threonine residues 1717:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 6 72 theme dissociation 718:729 arg1 fragmentation 731:743 higher energy collisional dissociation fragmentation 692:743 higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides 692:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 73 gly glycosylation 2172:2184 arg1 Srr1 2203:2206 Srr1 2203:2206 Srr1 2203:2206 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 1 74 theme Serine-rich 132:142 arg1 family 214:219 a family 212:219 a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci 212:309 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 1 74 theme Serine-rich 132:142 arg1 proteins 150:157 Serine-rich (Srr) proteins 132:157 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria 132:206 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 1 74 theme Serine-rich 132:142 arg1 Srr 145:147 Srr 145:147 Srr 145:147 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 6 75 theme energy 699:704 arg1 fragmentation 731:743 higher energy collisional dissociation fragmentation 692:743 higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides 692:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 0 76 theme Srr1 68:71 arg1 region 34:39 the N-terminal region 19:39 the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae 19:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 14 77 theme protein 2327:2333 arg1 region 2313:2318 the N-terminal region 2298:2318 the N-terminal region of the protein 2298:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 4 78 gly glycopeptides 567:579 arg2 glycopeptides 567:579 potential glycopeptides 557:579 potential glycopeptides 557:579 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 4 79 theme trypsin 538:544 arg1 digestion 546:554 trypsin digestion 538:554 trypsin digestion 538:554 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 6 80 theme higher 692:697 arg1 fragmentation 731:743 higher energy collisional dissociation fragmentation 692:743 higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides 692:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 10 81 theme Lectin 1366:1371 arg1 binding 1373:1379 Lectin binding 1366:1379 Lectin binding 1366:1379 Lectin binding and monosaccharide composition analysis strongly suggested that HexNAc and Hex correspond to N-acetylglucosamine and glucose, respectively. 24797265 0 82 theme Streptococcus 76:88 arg1 agalactiae 90:99 Streptococcus agalactiae 76:99 Streptococcus agalactiae 76:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 14 83 theme located 2287:2293 arg1 sites 2281:2285 six glycosylation sites 2263:2285 six glycosylation sites located in the N-terminal region of the protein 2263:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 84 theme spectrometry 2101:2112 arg1 techniques 2114:2123 fragmentation mass spectrometry techniques 2082:2123 fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein 2082:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 14 85 theme Srr1 2203:2206 arg1 heterogeneity 2186:2198 glycosylation heterogeneity 2172:2198 glycosylation heterogeneity of Srr1 2172:2206 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 13 86 theme glycosyltransferase 1862:1880 arg1 genes 1891:1895 accessory glycosyltransferase encoding genes 1852:1895 accessory glycosyltransferase encoding genes 1852:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 10 87 theme composition 1400:1410 arg1 analysis 1412:1419 monosaccharide composition analysis 1385:1419 monosaccharide composition analysis 1385:1419 Lectin binding and monosaccharide composition analysis strongly suggested that HexNAc and Hex correspond to N-acetylglucosamine and glucose, respectively. 24797265 0 88 theme mass 113:116 arg1 spectrometry 118:129 mass spectrometry 113:129 mass spectrometry 113:129 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 4 89 dep induced 610:616 arg1 collision 600:608 collision 600:608 collision 600:608 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 14 90 theme glycosylation 2267:2279 arg1 sites 2281:2285 six glycosylation sites 2263:2285 six glycosylation sites located in the N-terminal region of the protein 2263:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 6 91 theme simultaneous 762:773 arg1 loss 775:778 the simultaneous loss 758:778 the simultaneous loss of sugar residues 758:796 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 3 92 theme glycosylation 464:476 arg1 analysis 448:455 the first mass-spectrometry analysis 420:455 the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1 420:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 13 93 theme mutant 1829:1834 arg1 strains 1836:1842 mutant strains 1829:1842 mutant strains lacking accessory glycosyltransferase encoding genes 1829:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 11 94 theme glycans 1580:1586 arg1 glycans 1580:1586 glycans 1580:1586 glycans generating a wide structural diversity of Srr1 1580:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 11 94 theme glycans 1580:1586 arg1 variety 1569:1575 a variety 1567:1575 a variety of glycans generating a wide structural diversity of Srr1 1567:1633 The same protein segment can be modified with a variety of glycans generating a wide structural diversity of Srr1. 24797265 0 95 theme N-terminal 23:32 arg1 region 34:39 the N-terminal region 19:39 the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae 19:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 1 96 theme pathogenic 269:278 arg1 staphylococci 280:292 pathogenic staphylococci 269:292 pathogenic staphylococci 269:292 Serine-rich (Srr) proteins exposed at the surface of Gram-positive bacteria are a family of adhesins that contribute to the virulence of pathogenic staphylococci and streptococci. 24797265 4 97 theme dissociation 618:629 arg1 spectra 631:637 collision induced dissociation spectra 600:637 collision induced dissociation spectra using X! 600:646 After Srr1 enrichment and trypsin digestion, potential glycopeptides were identified in collision induced dissociation spectra using X! 24797265 9 98 theme same 1175:1178 arg1 residue 1188:1194 the same protein residue 1171:1194 the same protein residue 1171:1194 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 3 99 theme Srr1 506:509 arg1 glycosylation 464:476 the glycosylation 460:476 the glycosylation of Streptococcus agalactiae Srr1 460:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 0 100 gly O-Glycosylation 0:14 arg1 region 34:39 the N-terminal region 19:39 the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae 19:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 12 101 theme threonine 1771:1779 arg1 residues 1781:1788 one threonine residues 1767:1788 one threonine residues 1767:1788 Electron transfer dissociation was used to assign glycosylation sites leading to the unambiguous identification of six serines and one threonine residues. 24797265 3 102 theme mass-spectrometry 430:446 arg1 analysis 448:455 the first mass-spectrometry analysis 420:455 the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1 420:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 13 103 theme purified 1803:1810 arg1 Srr1 1812:1815 purified Srr1 1803:1815 purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes 1803:1895 Analysis of purified Srr1 produced in mutant strains lacking accessory glycosyltransferase encoding genes demonstrates that O-GlcNAcylation is an initial step in Srr1 glycosylation that is likely required for subsequent decoration with Hex. 24797265 6 104 theme ions 832:835 arg1 fragmentation 850:862 backbone fragmentation 841:862 backbone fragmentation for glycopeptides 841:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 6 104 theme ions 832:835 arg1 production 799:808 production 799:808 production of diagnostic oxonium ions 799:835 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 6 104 theme ions 832:835 arg1 loss 775:778 the simultaneous loss 758:778 the simultaneous loss of sugar residues 758:796 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 3 105 theme Streptococcus 481:493 arg1 Srr1 506:509 Streptococcus agalactiae Srr1 481:509 Streptococcus agalactiae Srr1 481:509 We report here the first mass-spectrometry analysis of the glycosylation of Streptococcus agalactiae Srr1. 24797265 0 106 theme serine-rich 48:58 arg1 Srr1 68:71 the serine-rich adhesin Srr1 44:71 the serine-rich adhesin Srr1 of Streptococcus agalactiae 44:99 O-Glycosylation of the N-terminal region of the serine-rich adhesin Srr1 of Streptococcus agalactiae explored by mass spectrometry. 24797265 6 107 theme diagnostic 813:822 arg1 ions 832:835 diagnostic oxonium ions 813:835 diagnostic oxonium ions 813:835 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 108 theme techniques 2114:2123 arg1 combination 2067:2077 a combination 2065:2077 a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein 2065:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 6 109 theme residues 789:796 arg1 fragmentation 850:862 backbone fragmentation 841:862 backbone fragmentation for glycopeptides 841:880 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 6 109 theme residues 789:796 arg1 production 799:808 production 799:808 production of diagnostic oxonium ions 799:835 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 6 109 theme residues 789:796 arg1 loss 775:778 the simultaneous loss 758:778 the simultaneous loss of sugar residues 758:796 The approach was then refined using higher energy collisional dissociation fragmentation which led to the simultaneous loss of sugar residues, production of diagnostic oxonium ions and backbone fragmentation for glycopeptides. 24797265 14 110 theme mass 2096:2099 arg1 techniques 2114:2123 fragmentation mass spectrometry techniques 2082:2123 fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein 2082:2333 In summary, our data obtained by a combination of fragmentation mass spectrometry techniques associated to a new software tool, demonstrate glycosylation heterogeneity of Srr1, characterize a new protein modification, and identify six glycosylation sites located in the N-terminal region of the protein. 24797265 9 111 theme glycan 1227:1232 arg1 modification 1297:1308 a novel modification 1289:1308 a novel modification that was identified as O-acetylated-N-acetylhexosamine 1289:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 111 theme glycan 1227:1232 arg1 hexose 1277:1282 hexose 1277:1282 hexose 1277:1282 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 111 theme glycan 1227:1232 arg1 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 N-acetylhexosamine 1257:1274 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 24797265 9 111 theme glycan 1227:1232 arg1 combinations 1234:1245 different glycan combinations 1217:1245 different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine 1217:1363 Our data unambiguously indicate that the same protein residue can be modified with different glycan combinations including N-acetylhexosamine, hexose, and a novel modification that was identified as O-acetylated-N-acetylhexosamine. 29131431 0 0 theme New 78:80 arg1 Agents 96:101 New Anticoagulant Agents 78:101 New Anticoagulant Agents 78:101 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 4 1 theme facile 528:533 arg1 method 535:540 facile method 528:540 facile method 528:540 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 6 2 theme assay 1089:1093 arg1 activities 1095:1104 assay activities 1089:1104 assay activities 1089:1104 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 8 3 theme most 1387:1390 arg1 potency 1392:1398 potency 1392:1398 potency 1392:1398 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 8 3 theme most 1387:1390 arg1 consistent 1414:1423 consistent 1414:1423 consistent 1414:1423 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 4 4 theme FuCS 611:614 arg1 unit 603:606 the repeating trisaccharide unit 575:606 the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers 575:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 6 5 dep time 1056:1059 arg1 activities 1095:1104 assay activities 1089:1104 assay activities 1089:1104 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 8 6 with glycoclusters 1316:1328 arg1 residue 1365:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 7 7 theme intrinsic 1199:1207 arg1 activities 1223:1232 intrinsic anticoagulant activities 1199:1232 intrinsic anticoagulant activities 1199:1232 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 7 8 theme Further 1107:1113 arg1 tests 1132:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 9 9 theme novel 1588:1592 arg1 agents 1608:1613 novel anticoagulant agents 1588:1613 novel anticoagulant agents 1588:1613 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 9 10 theme linear 1513:1518 arg1 glycosaminoglycans 1520:1537 linear glycosaminoglycans 1513:1537 linear glycosaminoglycans 1513:1537 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 5 11 theme branches 766:773 arg1 series 676:681 A series 674:681 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs 674:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 7 12 contain had 1239:1241 arg2 effect 1250:1255 little effect 1243:1255 little effect 1243:1255 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 7 12 contain had 1239:1241 arg1 33 b-40 b 1167:1175 glycoclusters 33 b-40 b 1153:1175 glycoclusters 33 b-40 b 1153:1175 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 5 13 theme simplified 686:695 arg1 glycomimetics 702:714 simplified FuCS glycomimetics 686:714 simplified FuCS glycomimetics that have highly tunable structures 686:750 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 7 14 theme coagulation 1285:1295 arg1 pathways 1297:1304 the extrinsic and common coagulation pathways 1260:1304 the extrinsic and common coagulation pathways 1260:1304 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 0 15 theme Anticoagulant 82:94 arg1 Agents 96:101 New Anticoagulant Agents 78:101 New Anticoagulant Agents 78:101 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 5 16 theme FuCS 697:700 arg1 glycomimetics 702:714 simplified FuCS glycomimetics 686:714 simplified FuCS glycomimetics that have highly tunable structures 686:750 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 6 17 theme Remarkable 868:877 arg1 branches 976:983 the branches 972:983 the branches increased 972:993 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 6 17 theme Remarkable 868:877 arg1 improvement 879:889 Remarkable improvement 868:889 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities 868:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 2 18 theme anticoagulant 302:314 arg1 mechanism 316:324 a different anticoagulant mechanism 290:324 a different anticoagulant mechanism 290:324 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 8 19 theme fucosyl 1357:1363 arg1 residue 1365:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 2 20 theme decreased 365:373 arg1 risks 375:379 decreased risks 365:379 decreased risks of adverse effects and bleeding 365:411 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 7 21 theme little 1243:1248 arg1 effect 1250:1255 little effect 1243:1255 little effect 1243:1255 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 4 22 theme sulfate 656:662 arg1 polymers 664:671 chondroitin sulfate polymers 644:671 chondroitin sulfate polymers 644:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 2 23 theme different 292:300 arg1 mechanism 316:324 a different anticoagulant mechanism 290:324 a different anticoagulant mechanism 290:324 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 9 24 theme new 1551:1553 arg1 framework 1555:1563 a new framework 1549:1563 a new framework for the development of novel anticoagulant agents 1549:1613 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 1 25 theme distinct 161:168 arg1 glycosaminoglycan 170:186 a structurally distinct glycosaminoglycan 146:186 a structurally distinct glycosaminoglycan with excellent anticoagulant activity 146:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 1 25 theme distinct 161:168 arg1 sulfate 128:134 Fucosylated chondroitin sulfate 104:134 Fucosylated chondroitin sulfate (FuCS) 104:141 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 5 26 theme defined 780:786 arg1 motifs 798:803 defined sulfation motifs 780:803 defined sulfation motifs 780:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 7 27 theme glycoclusters 1153:1165 arg1 33 b-40 b 1167:1175 glycoclusters 33 b-40 b 1153:1175 glycoclusters 33 b-40 b 1153:1175 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 6 28 theme prothrombin 1044:1054 arg1 PT 1062:1063 PT 1062:1063 PT 1062:1063 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 6 28 theme prothrombin 1044:1054 arg1 time 1056:1059 prothrombin time 1044:1059 prothrombin time (PT) 1044:1064 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 2 29 theme heparin 339:345 arg1 derivatives 347:357 heparin derivatives 339:357 heparin derivatives 339:357 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 2 30 theme bleeding 404:411 arg1 risks 375:379 decreased risks 365:379 decreased risks of adverse effects and bleeding 365:411 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 4 31 theme repeating 579:587 arg1 unit 603:606 the repeating trisaccharide unit 575:606 the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers 575:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 8 32 with consistent 1414:1423 arg1 that 1430:1433 that 1430:1433 that 1430:1433 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 8 33 theme 2,4-di-O-sulfated 1339:1355 arg1 residue 1365:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 the 2,4-di-O-sulfated fucosyl residue 1335:1371 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 1 34 theme excellent 193:201 arg1 activity 217:224 excellent anticoagulant activity 193:224 excellent anticoagulant activity 193:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 0 35 theme Chondroitin 25:35 arg1 Sulfate 37:43 Fucosylated Chondroitin Sulfate 13:43 Fucosylated Chondroitin Sulfate 13:43 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 5 36 theme tunable 733:739 arg1 structures 741:750 highly tunable structures 726:750 highly tunable structures 726:750 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 1 37 theme anticoagulant 203:215 arg1 activity 217:224 excellent anticoagulant activity 193:224 excellent anticoagulant activity 193:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 0 38 theme Fucosylated 13:23 arg1 Sulfate 37:43 Fucosylated Chondroitin Sulfate 13:43 Fucosylated Chondroitin Sulfate 13:43 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 4 39 theme unit 603:606 arg1 synthesis 562:570 the synthesis 558:570 the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers 558:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 7 40 theme inhibition 1121:1130 arg1 tests 1132:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 6 41 theme assay 939:943 arg1 activities 945:954 activated partial thromboplastin time (APTT) assay activities 894:954 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 1 42 theme Fucosylated 104:114 arg1 FuCS 137:140 FuCS 137:140 FuCS 137:140 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 1 42 theme Fucosylated 104:114 arg1 sulfate 128:134 Fucosylated chondroitin sulfate 104:134 Fucosylated chondroitin sulfate (FuCS) 104:141 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 1 42 theme Fucosylated 104:114 arg1 glycosaminoglycan 170:186 a structurally distinct glycosaminoglycan 146:186 a structurally distinct glycosaminoglycan with excellent anticoagulant activity 146:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 3 43 theme defined 494:500 arg1 oligosaccharides 502:517 structurally defined oligosaccharides 481:517 structurally defined oligosaccharides 481:517 However, further exploitation has been hindered by the scarcity of structurally defined oligosaccharides. 29131431 4 44 theme trisaccharide 589:601 arg1 unit 603:606 the repeating trisaccharide unit 575:606 the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers 575:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 6 45 from improvement 879:889 arg1 activities 945:954 activated partial thromboplastin time (APTT) assay activities 894:954 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 7 46 theme anticoagulant 1209:1221 arg1 activities 1223:1232 intrinsic anticoagulant activities 1199:1232 intrinsic anticoagulant activities 1199:1232 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 0 47 theme Sulfate 37:43 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Fucosylated Chondroitin Sulfate 0:43 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 5 48 theme controllable 753:764 arg1 branches 766:773 controllable branches 753:773 controllable branches 753:773 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 6 49 theme thrombin 1070:1077 arg1 TT 1085:1086 TT 1085:1086 TT 1085:1086 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 6 49 theme thrombin 1070:1077 arg1 time 1079:1082 thrombin time 1070:1082 thrombin time (TT) 1070:1087 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 7 50 theme extrinsic 1264:1272 arg1 pathways 1297:1304 the extrinsic and common coagulation pathways 1260:1304 the extrinsic and common coagulation pathways 1260:1304 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 5 51 theme alkyne-azide 840:851 arg1 cycloaddition 853:865 copper-catalyzed alkyne-azide cycloaddition 823:865 copper-catalyzed alkyne-azide cycloaddition 823:865 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 52 contain have 721:724 arg1 motifs 798:803 defined sulfation motifs 780:803 defined sulfation motifs 780:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 52 contain have 721:724 arg2 structures 741:750 highly tunable structures 726:750 highly tunable structures 726:750 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 52 contain have 721:724 arg1 glycomimetics 702:714 simplified FuCS glycomimetics 686:714 simplified FuCS glycomimetics that have highly tunable structures 686:750 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 52 contain have 721:724 arg1 branches 766:773 controllable branches 753:773 controllable branches 753:773 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 53 theme glycomimetics 702:714 arg1 series 676:681 A series 674:681 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs 674:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 5 54 theme motifs 798:803 arg1 series 676:681 A series 674:681 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs 674:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 0 55 dep Glycoclusters 45:57 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Fucosylated Chondroitin Sulfate 0:43 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 0 55 dep Glycoclusters 45:57 arg1 Route 69:73 A Robust Route 60:73 A Robust Route to New Anticoagulant Agents 60:101 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 1 56 theme chondroitin 116:126 arg1 FuCS 137:140 FuCS 137:140 FuCS 137:140 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 1 56 theme chondroitin 116:126 arg1 sulfate 128:134 Fucosylated chondroitin sulfate 104:134 Fucosylated chondroitin sulfate (FuCS) 104:141 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 1 56 theme chondroitin 116:126 arg1 glycosaminoglycan 170:186 a structurally distinct glycosaminoglycan 146:186 a structurally distinct glycosaminoglycan with excellent anticoagulant activity 146:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 3 57 theme further 423:429 arg1 exploitation 431:442 further exploitation 423:442 further exploitation 423:442 However, further exploitation has been hindered by the scarcity of structurally defined oligosaccharides. 29131431 7 58 theme FXase 1115:1119 arg1 tests 1132:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests 1107:1136 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 6 59 theme time 927:930 arg1 activities 945:954 activated partial thromboplastin time (APTT) assay activities 894:954 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 7 60 theme common 1278:1283 arg1 pathways 1297:1304 the extrinsic and common coagulation pathways 1260:1304 the extrinsic and common coagulation pathways 1260:1304 Further FXase inhibition tests suggested that glycoclusters 33 b-40 b selectively inhibited intrinsic anticoagulant activities, but had little effect on the extrinsic and common coagulation pathways. 29131431 9 61 theme anticoagulant 1594:1606 arg1 agents 1608:1613 novel anticoagulant agents 1588:1613 novel anticoagulant agents 1588:1613 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 9 62 theme FuCS 1469:1472 arg1 clusters 1474:1481 These FuCS clusters 1463:1481 These FuCS clusters 1463:1481 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 6 63 theme thromboplastin 912:925 arg1 APTT 933:936 APTT 933:936 APTT 933:936 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 6 63 theme thromboplastin 912:925 arg1 time 927:930 partial thromboplastin time 904:930 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 3 64 theme oligosaccharides 502:517 arg1 scarcity 469:476 the scarcity 465:476 the scarcity of structurally defined oligosaccharides 465:517 However, further exploitation has been hindered by the scarcity of structurally defined oligosaccharides. 29131431 8 65 theme natural 1438:1444 arg1 polysaccharides 1446:1460 natural polysaccharides 1438:1460 natural polysaccharides 1438:1460 Notably, glycoclusters with the 2,4-di-O-sulfated fucosyl residue displayed the most potency, which was in consistent with that of natural polysaccharides. 29131431 1 66 with glycosaminoglycan 170:186 arg1 activity 217:224 excellent anticoagulant activity 193:224 excellent anticoagulant activity 193:224 Fucosylated chondroitin sulfate (FuCS) is a structurally distinct glycosaminoglycan with excellent anticoagulant activity. 29131431 6 67 theme partial 904:910 arg1 APTT 933:936 APTT 933:936 APTT 933:936 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 6 67 theme partial 904:910 arg1 time 927:930 partial thromboplastin time 904:930 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 9 68 theme agents 1608:1613 arg1 development 1573:1583 the development 1569:1583 the development of novel anticoagulant agents 1569:1613 These FuCS clusters demonstrated potency to mimic linear glycosaminoglycans and offer a new framework for the development of novel anticoagulant agents. 29131431 5 69 theme sulfation 788:796 arg1 motifs 798:803 defined sulfation motifs 780:803 defined sulfation motifs 780:803 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 6 70 theme activated 894:902 arg1 activities 945:954 activated partial thromboplastin time (APTT) assay activities 894:954 activated partial thromboplastin time (APTT) assay activities 894:954 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29131431 2 71 theme depolymerized 258:270 arg1 fragments 272:280 its depolymerized fragments 254:280 its depolymerized fragments 254:280 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 4 72 theme polymers 664:671 arg1 degradation 629:639 the degradation 625:639 the degradation of chondroitin sulfate polymers 625:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 2 73 theme effects 392:398 arg1 risks 375:379 decreased risks 365:379 decreased risks of adverse effects and bleeding 365:411 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 4 74 theme chondroitin 644:654 arg1 polymers 664:671 chondroitin sulfate polymers 644:671 chondroitin sulfate polymers 644:671 Herein, facile method is reported for the synthesis of the repeating trisaccharide unit of FuCS based on the degradation of chondroitin sulfate polymers. 29131431 0 75 theme Robust 62:67 arg1 Route 69:73 A Robust Route 60:73 A Robust Route to New Anticoagulant Agents 60:101 Synthesis of Fucosylated Chondroitin Sulfate Glycoclusters: A Robust Route to New Anticoagulant Agents. 29131431 5 76 theme copper-catalyzed 823:838 arg1 cycloaddition 853:865 copper-catalyzed alkyne-azide cycloaddition 823:865 copper-catalyzed alkyne-azide cycloaddition 823:865 A series of simplified FuCS glycomimetics that have highly tunable structures, controllable branches, and defined sulfation motifs were generated by copper-catalyzed alkyne-azide cycloaddition. 29131431 2 77 theme adverse 384:390 arg1 effects 392:398 adverse effects 384:398 adverse effects 384:398 Studies show that FuCS and its depolymerized fragments exhibit a different anticoagulant mechanism from that of heparin derivatives, with decreased risks of adverse effects and bleeding. 29131431 6 78 theme significant 1003:1013 arg1 influences 1015:1024 no significant influences 1000:1024 no significant influences 1000:1024 Remarkable improvement in activated partial thromboplastin time (APTT) assay activities was observed as the branches increased, but no significant influences were observed for prothrombin time (PT) and thrombin time (TT) assay activities. 29065342 5 0 link O-linked 873:880 arg1 LeX 882:884 O-linked LeX 873:884 O-linked LeX 873:884 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 1 1 theme critical 219:226 arg1 roles 228:232 critical roles 219:232 critical roles 219:232 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 0 2 theme LewisX 70:75 arg1 glycopeptides 77:89 O-linked LewisX glycopeptides 61:89 O-linked LewisX glycopeptides 61:89 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 1 3 gly glycoproteins 195:207 arg1 glycoproteins 195:207 many cell surface glycoproteins 177:207 many cell surface glycoproteins 177:207 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 1 4 theme Fucα1→3 141:147 arg1 LewisX 92:97 LewisX 92:97 LewisX (LeX) 92:103 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 1 4 theme Fucα1→3 141:147 arg1 GlcNAc 149:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc 108:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses 108:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 4 5 gly glycopeptides 747:759 arg2 glycopeptides 747:759 diverse O-linked LeX glycopeptides 726:759 diverse O-linked LeX glycopeptides 726:759 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 3 6 theme regioselective 483:496 arg1 anhydride 535:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride 481:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation 481:574 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 4 7 theme Fmoc-threonine 596:609 arg1 acid 617:620 an Fmoc-threonine amino acid 593:620 an Fmoc-threonine amino acid 593:620 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 4 8 link O-linked 734:741 arg1 glycopeptides 747:759 diverse O-linked LeX glycopeptides 726:759 diverse O-linked LeX glycopeptides 726:759 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 1 9 theme innate 237:242 arg1 responses 264:272 innate and adaptive immune responses 237:272 innate and adaptive immune responses 237:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 4 10 theme diverse 726:732 arg1 glycopeptides 747:759 diverse O-linked LeX glycopeptides 726:759 diverse O-linked LeX glycopeptides 726:759 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 3 11 theme piperidine/triflic 516:533 arg1 anhydride 535:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride 481:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation 481:574 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 4 12 theme phase 680:684 arg1 synthesis 694:702 solid phase peptide synthesis 674:702 solid phase peptide synthesis 674:702 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 5 13 theme described 766:774 arg1 approach 776:783 The described approach 762:783 The described approach 762:783 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 13 theme described 766:774 arg1 applicable 796:805 applicable 796:805 applicable 796:805 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 14 gly glycopeptides 848:860 arg2 glycopeptides 848:860 complex glycopeptides 840:860 complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 840:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 0 15 gly glycopeptides 77:89 arg2 glycopeptides 77:89 O-linked LewisX glycopeptides 61:89 O-linked LewisX glycopeptides 61:89 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 4 16 theme acid 617:620 arg1 presence 581:588 The presence 577:588 The presence of an Fmoc-threonine amino acid 577:620 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 2 17 theme structure-function 364:381 arg1 studies 383:389 structure-function studies 364:389 structure-function studies of the LeX determinant 364:412 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 3 18 theme 1-benzenesulfinyl 498:514 arg1 anhydride 535:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride 481:543 a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation 481:574 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 4 19 theme solid 674:678 arg1 synthesis 694:702 solid phase peptide synthesis 674:702 solid phase peptide synthesis 674:702 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 2 20 theme efficient 284:292 arg1 synthesis 294:302 efficient synthesis 284:302 efficient synthesis of glycopeptides bearing LeX 284:331 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 5 21 theme variety 829:835 arg1 synthesis 814:822 the synthesis 810:822 the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 810:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 4 22 theme O-linked 734:741 arg1 glycopeptides 747:759 diverse O-linked LeX glycopeptides 726:759 diverse O-linked LeX glycopeptides 726:759 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 5 23 contain containing 862:871 arg2 sLeX 904:907 sLeX 904:907 sLeX 904:907 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 23 contain containing 862:871 arg1 glycopeptides 848:860 complex glycopeptides 840:860 complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 840:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 23 contain containing 862:871 arg2 LewisX 896:901 sialyl LewisX 889:901 sialyl LewisX (sLeX) 889:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 23 contain containing 862:871 arg2 LeX 882:884 O-linked LeX 873:884 O-linked LeX 873:884 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 24 theme sialyl 889:894 arg1 sLeX 904:907 sLeX 904:907 sLeX 904:907 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 24 theme sialyl 889:894 arg1 LewisX 896:901 sialyl LewisX 889:901 sialyl LewisX (sLeX) 889:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 4 25 theme peptide 686:692 arg1 synthesis 694:702 solid phase peptide synthesis 674:702 solid phase peptide synthesis 674:702 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 0 26 theme threonine 29:37 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 0:90 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 1 27 theme adaptive 248:255 arg1 responses 264:272 innate and adaptive immune responses 237:272 innate and adaptive immune responses 237:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 2 28 theme determinant 402:412 arg1 studies 383:389 structure-function studies 364:389 structure-function studies of the LeX determinant 364:412 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 0 29 theme LewisX-O-Core-1 13:27 arg1 threonine 29:37 LewisX-O-Core-1 threonine 13:37 LewisX-O-Core-1 threonine 13:37 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 1 30 theme immune 257:262 arg1 responses 264:272 innate and adaptive immune responses 237:272 innate and adaptive immune responses 237:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 2 31 theme major 343:347 arg1 limitation 349:358 a major limitation 341:358 a major limitation for structure-function studies of the LeX determinant 341:412 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 2 32 theme LeX 398:400 arg1 determinant 402:412 the LeX determinant 394:412 the LeX determinant 394:412 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 5 33 theme O-linked 873:880 arg1 LeX 882:884 O-linked LeX 873:884 O-linked LeX 873:884 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 4 34 theme pentasaccharide 655:669 arg1 incorporation 634:646 incorporation 634:646 incorporation of the pentasaccharide in solid phase peptide synthesis 634:702 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 1 35 theme many 177:180 arg1 glycoproteins 195:207 many cell surface glycoproteins 177:207 many cell surface glycoproteins 177:207 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 4 36 from incorporation 634:646 arg1 synthesis 694:702 solid phase peptide synthesis 674:702 solid phase peptide synthesis 674:702 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 1 37 theme cell 182:185 arg1 glycoproteins 195:207 many cell surface glycoproteins 177:207 many cell surface glycoproteins 177:207 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 2 38 gly glycopeptides 307:319 arg2 glycopeptides 307:319 glycopeptides 307:319 glycopeptides bearing LeX 307:331 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 3 39 theme pentasaccharide 457:471 arg1 synthesis 438:446 a total synthesis 430:446 a total synthesis of a LeX pentasaccharide 1 430:473 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 0 40 theme building 42:49 arg1 block 51:55 A building block 40:55 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 0:90 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 0 41 link O-linked 61:68 arg1 glycopeptides 77:89 O-linked LewisX glycopeptides 61:89 O-linked LewisX glycopeptides 61:89 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 2 42 theme glycopeptides 307:319 arg1 synthesis 294:302 efficient synthesis 284:302 efficient synthesis of glycopeptides bearing LeX 284:331 However, efficient synthesis of glycopeptides bearing LeX remains a major limitation for structure-function studies of the LeX determinant. 29065342 4 43 theme LeX 743:745 arg1 glycopeptides 747:759 diverse O-linked LeX glycopeptides 726:759 diverse O-linked LeX glycopeptides 726:759 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 1 44 theme surface 187:193 arg1 glycoproteins 195:207 many cell surface glycoproteins 177:207 many cell surface glycoproteins 177:207 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 5 45 theme complex 840:846 arg1 glycopeptides 848:860 complex glycopeptides 840:860 complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 840:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 1 46 theme branched 110:117 arg1 LewisX 92:97 LewisX 92:97 LewisX (LeX) 92:103 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 1 46 theme branched 110:117 arg1 GlcNAc 149:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc 108:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses 108:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 4 47 theme amino 611:615 arg1 acid 617:620 an Fmoc-threonine amino acid 593:620 an Fmoc-threonine amino acid 593:620 The presence of an Fmoc-threonine amino acid facilitates incorporation of the pentasaccharide in solid phase peptide synthesis, providing a route to diverse O-linked LeX glycopeptides. 29065342 0 48 theme O-linked 61:68 arg1 glycopeptides 77:89 O-linked LewisX glycopeptides 61:89 O-linked LewisX glycopeptides 61:89 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 3 49 theme total 432:436 arg1 synthesis 438:446 a total synthesis 430:446 a total synthesis of a LeX pentasaccharide 1 430:473 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 5 50 theme glycopeptides 848:860 arg1 variety 829:835 a variety 827:835 a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 827:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 5 50 theme glycopeptides 848:860 arg1 glycopeptides 848:860 complex glycopeptides 840:860 complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX) 840:908 The described approach is broadly applicable to the synthesis of a variety of complex glycopeptides containing O-linked LeX or sialyl LewisX (sLeX). 29065342 1 51 theme trisaccharide 119:131 arg1 LewisX 92:97 LewisX 92:97 LewisX (LeX) 92:103 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 1 51 theme trisaccharide 119:131 arg1 GlcNAc 149:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc 108:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses 108:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 3 52 theme LeX 453:455 arg1 pentasaccharide 457:471 a LeX pentasaccharide 1 451:473 a LeX pentasaccharide 1 451:473 Here we report a total synthesis of a LeX pentasaccharide 1 using a regioselective 1-benzenesulfinyl piperidine/triflic anhydride promoted [3 + 2] glycosylation. 29065342 0 53 dep Synthesis 0:8 arg1 block 51:55 A building block 40:55 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 0:90 Synthesis of LewisX-O-Core-1 threonine: A building block for O-linked LewisX glycopeptides. 29065342 1 54 theme Galβ1→4 133:139 arg1 LewisX 92:97 LewisX 92:97 LewisX (LeX) 92:103 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 29065342 1 54 theme Galβ1→4 133:139 arg1 GlcNAc 149:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc 108:154 a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses 108:272 LewisX (LeX) is a branched trisaccharide Galβ1→4(Fucα1→3)GlcNAc that is expressed on many cell surface glycoproteins and plays critical roles in innate and adaptive immune responses. 25851037 4 0 theme intravenous 995:1005 arg1 immunoglobulin 1007:1020 intravenous immunoglobulin 995:1020 intravenous immunoglobulin (IVIG) 995:1027 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 0 theme intravenous 995:1005 arg1 product 1038:1044 a blood product 1030:1044 a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1030:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 0 theme intravenous 995:1005 arg1 IVIG 1023:1026 IVIG 1023:1026 IVIG 1023:1026 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 1 contain containing 1046:1055 arg1 immunoglobulin 1007:1020 intravenous immunoglobulin 995:1020 intravenous immunoglobulin (IVIG) 995:1027 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 1 contain containing 1046:1055 arg2 antibodies 1079:1088 pooled polyvalent IgG antibodies 1057:1088 pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1057:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 1 contain containing 1046:1055 arg1 product 1038:1044 a blood product 1030:1044 a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1030:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 2 theme glycoforms 1300:1309 arg1 subset 1290:1295 a subset 1288:1295 a subset of glycoforms 1288:1309 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 2 3 theme detailed 354:361 arg1 description 363:373 detailed description 354:373 detailed description of glycan structures 354:394 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 4 4 theme blood 1032:1036 arg1 immunoglobulin 1007:1020 intravenous immunoglobulin 995:1020 intravenous immunoglobulin (IVIG) 995:1027 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 4 theme blood 1032:1036 arg1 product 1038:1044 a blood product 1030:1044 a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1030:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 5 from lectins 1267:1273 arg1 readouts 1244:1251 readouts 1244:1251 readouts from multiple lectins 1244:1273 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 6 theme intact 978:983 arg1 IgGs 985:988 intact IgGs 978:988 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 5 7 with findings 1158:1165 arg1 profiling 1196:1204 industry-standard LC-MS profiling 1172:1204 industry-standard LC-MS profiling 1172:1204 We corroborate our findings with industry-standard LC-MS profiling. 25851037 4 8 theme terminal 876:883 arg1 moieties 892:899 terminal glycan moieties 876:899 terminal glycan moieties 876:899 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 9 theme pooled 1057:1062 arg1 antibodies 1079:1088 pooled polyvalent IgG antibodies 1057:1088 pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1057:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 10 theme Fc 955:956 arg1 proteins 958:965 in vitro and in-cell glycan-engineered Fc proteins 916:965 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 11 theme glycan 885:890 arg1 moieties 892:899 terminal glycan moieties 876:899 terminal glycan moieties 876:899 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 12 theme immunosorbent 835:847 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 12 theme immunosorbent 835:847 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 3 13 from probes 533:538 arg1 microarrays 543:553 microarrays 543:553 microarrays 543:553 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 4 14 theme glycan-engineered 937:953 arg1 proteins 958:965 in vitro and in-cell glycan-engineered Fc proteins 916:965 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 3 15 theme readers 722:728 arg1 capabilities 685:696 the capabilities 681:696 the capabilities of automated microarray readers 681:728 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 1 16 with biotherapeutics 145:159 arg1 carbohydrates 167:179 carbohydrates 167:179 carbohydrates playing an important role for therapeutic efficacy 167:230 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 6 17 theme glycan 1561:1566 arg1 analysis 1568:1575 quantitative glycan analysis 1548:1575 quantitative glycan analysis 1548:1575 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 6 18 theme epitopes 1411:1418 arg1 levels 1401:1406 levels 1401:1406 levels of epitopes 1401:1418 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 19 theme human 1125:1129 arg1 donors 1131:1136 healthy human donors 1117:1136 healthy human donors 1117:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 20 dep in 916:917 arg1 vitro 919:923 vitro 919:923 vitro 919:923 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 3 21 theme automated 701:709 arg1 readers 722:728 automated microarray readers 701:728 automated microarray readers 701:728 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 0 22 theme Sialylated 36:45 arg1 Analyses 57:64 Sialylated Glycoform Analyses 36:64 Sialylated Glycoform Analyses Using Lectin Complexes 36:87 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes. 25851037 6 23 theme comparable 1437:1446 arg1 sensitivities 1423:1435 sensitivities 1423:1435 sensitivities comparable to MS. Extendable to other biologics 1423:1483 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 24 link lectin-enzyme-linked 814:833 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 24 link lectin-enzyme-linked 814:833 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 0 25 theme Quantitative 2:13 arg1 Assay 26:30 A Quantitative Microtiter Assay 0:30 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes 0:87 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes. 25851037 3 26 theme microarray 711:720 arg1 readers 722:728 automated microarray readers 701:728 automated microarray readers 701:728 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 2 27 theme structures 385:394 arg1 description 363:373 detailed description 354:373 detailed description of glycan structures 354:394 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 5 28 theme industry-standard 1172:1188 arg1 profiling 1196:1204 industry-standard LC-MS profiling 1172:1204 industry-standard LC-MS profiling 1172:1204 We corroborate our findings with industry-standard LC-MS profiling. 25851037 6 29 theme multiple 1258:1265 arg1 lectins 1267:1273 multiple lectins 1258:1273 multiple lectins 1258:1273 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 2 30 theme glycan 378:383 arg1 structures 385:394 glycan structures 378:394 glycan structures 378:394 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 6 31 theme MS. 1451:1453 arg1 Extendable 1455:1464 MS. Extendable 1451:1464 MS. Extendable to other biologics 1451:1483 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 32 theme microtiter 796:805 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 32 theme microtiter 796:805 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 33 theme single- 1348:1354 arg1 glycosylation 1372:1384 single- versus dual-arm glycosylation 1348:1384 single- versus dual-arm glycosylation 1348:1384 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 34 theme lectin-enzyme-linked 814:833 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 34 theme lectin-enzyme-linked 814:833 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 2 35 theme mass 314:317 arg1 MS 333:334 MS 333:334 MS 333:334 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 35 theme mass 314:317 arg1 spectrometry 319:330 mass spectrometry 314:330 mass spectrometry (MS) 314:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 0 36 theme Microtiter 15:24 arg1 Assay 26:30 A Quantitative Microtiter Assay 0:30 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes 0:87 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes. 25851037 3 37 theme data 734:737 arg1 analysis 739:746 data analysis 734:746 data analysis 734:746 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 4 38 theme in 916:917 arg1 proteins 958:965 in vitro and in-cell glycan-engineered Fc proteins 916:965 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 1 39 theme structures 109:118 arg1 requirement 129:139 a key requirement 123:139 a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy 123:230 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 1 39 theme structures 109:118 arg1 Fidelity 90:97 Fidelity 90:97 Fidelity of glycan structures 90:118 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 4 40 theme healthy 1117:1123 arg1 donors 1131:1136 healthy human donors 1117:1136 healthy human donors 1117:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 2 41 theme data 478:481 arg1 sets 483:486 the considerable data sets 461:486 the considerable data sets they generate 461:500 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 6 42 theme quantitative 1548:1559 arg1 analysis 1568:1575 quantitative glycan analysis 1548:1575 quantitative glycan analysis 1548:1575 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 4 43 theme in-cell 929:935 arg1 proteins 958:965 in vitro and in-cell glycan-engineered Fc proteins 916:965 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 3 44 theme analyte 643:649 arg1 labeling 651:658 analyte labeling 643:658 analyte labeling to take advantage of the capabilities of automated microarray readers 643:728 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 2 45 theme profiling 254:262 arg1 chromatography 290:303 liquid chromatography 283:303 liquid chromatography (LC) 283:308 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 45 theme profiling 254:262 arg1 spectrometry 319:330 mass spectrometry 314:330 mass spectrometry (MS) 314:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 45 theme profiling 254:262 arg1 techniques 264:273 Comprehensive glycan profiling techniques 233:273 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS) 233:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 1 46 theme important 192:200 arg1 role 202:205 an important role 189:205 an important role for therapeutic efficacy 189:230 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 1 47 theme therapeutic 211:221 arg1 efficacy 223:230 therapeutic efficacy 211:230 therapeutic efficacy 211:230 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 2 48 theme considerable 465:476 arg1 sets 483:486 the considerable data sets 461:486 the considerable data sets they generate 461:500 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 4 49 from donors 1131:1136 arg1 plasma 1105:1110 plasma 1105:1110 plasma from healthy human donors 1105:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 2 50 theme glycan 247:252 arg1 chromatography 290:303 liquid chromatography 283:303 liquid chromatography (LC) 283:308 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 50 theme glycan 247:252 arg1 spectrometry 319:330 mass spectrometry 314:330 mass spectrometry (MS) 314:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 50 theme glycan 247:252 arg1 techniques 264:273 Comprehensive glycan profiling techniques 233:273 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS) 233:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 51 theme glycan 405:410 arg1 cleavage 412:419 glycan cleavage 405:419 glycan cleavage 405:419 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 4 52 from immunoglobulin 1007:1020 arg1 proteins 958:965 in vitro and in-cell glycan-engineered Fc proteins 916:965 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 52 from immunoglobulin 1007:1020 arg1 IgGs 985:988 intact IgGs 978:988 in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 916:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 5 53 theme LC-MS 1190:1194 arg1 profiling 1196:1204 industry-standard LC-MS profiling 1172:1204 industry-standard LC-MS profiling 1172:1204 We corroborate our findings with industry-standard LC-MS profiling. 25851037 2 54 theme Comprehensive 233:245 arg1 chromatography 290:303 liquid chromatography 283:303 liquid chromatography (LC) 283:308 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 54 theme Comprehensive 233:245 arg1 spectrometry 319:330 mass spectrometry 314:330 mass spectrometry (MS) 314:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 54 theme Comprehensive 233:245 arg1 techniques 264:273 Comprehensive glycan profiling techniques 233:273 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS) 233:335 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 3 55 used used 574:577 arg2 lectins 522:528 lectins 522:528 lectins as probes on microarrays 522:553 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 0 56 theme Glycoform 47:55 arg1 Analyses 57:64 Sialylated Glycoform Analyses 36:64 Sialylated Glycoform Analyses Using Lectin Complexes 36:87 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes. 25851037 3 57 dep in 608:609 arg1 situ 611:614 situ 611:614 situ 611:614 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 2 58 dep sets 483:486 arg1 generate 493:500 generate 493:500 generate 493:500 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 4 59 theme assay 849:853 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 59 theme assay 849:853 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 60 theme IgG 1075:1077 arg1 antibodies 1079:1088 pooled polyvalent IgG antibodies 1057:1088 pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1057:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 61 used used 1504:1507 arg2 ELISA 1491:1495 this ELISA 1486:1495 this ELISA 1486:1495 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 3 62 theme in 608:609 arg1 glycoprofiling 616:629 in situ glycoprofiling 608:629 in situ glycoprofiling 608:629 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 2 63 theme liquid 283:288 arg1 LC 306:307 LC 306:307 LC 306:307 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 2 63 theme liquid 283:288 arg1 chromatography 290:303 liquid chromatography 283:303 liquid chromatography (LC) 283:308 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 3 64 theme orthogonal 582:591 arg1 approaches 593:602 orthogonal approaches 582:602 orthogonal approaches for in situ glycoprofiling 582:629 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 1 65 theme key 125:127 arg1 requirement 129:139 a key requirement 123:139 a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy 123:230 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 1 65 theme key 125:127 arg1 Fidelity 90:97 Fidelity 90:97 Fidelity of glycan structures 90:118 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 1 66 theme glycan 102:107 arg1 structures 109:118 glycan structures 102:118 glycan structures 102:118 Fidelity of glycan structures is a key requirement for biotherapeutics, with carbohydrates playing an important role for therapeutic efficacy. 25851037 4 67 theme applicable 902:911 arg1 moieties 892:899 terminal glycan moieties 876:899 terminal glycan moieties 876:899 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 68 theme customizable 1213:1224 arg1 ELISA 1227:1231 This "customizable" ELISA 1207:1231 This "customizable" ELISA 1207:1231 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 0 69 theme Lectin 72:77 arg1 Complexes 79:87 Lectin Complexes 72:87 Lectin Complexes 72:87 A Quantitative Microtiter Assay for Sialylated Glycoform Analyses Using Lectin Complexes. 25851037 6 70 theme other 1469:1473 arg1 biologics 1475:1483 other biologics 1469:1483 other biologics 1469:1483 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 2 71 dep require 397:403 arg1 providing 344:352 providing 344:352 providing detailed description of glycan structures 344:394 Comprehensive glycan profiling techniques such as liquid chromatography (LC) and mass spectrometry (MS), while providing detailed description of glycan structures, require glycan cleavage, labeling, and paradigms to deconvolute the considerable data sets they generate. 25851037 3 72 theme other 510:514 arg1 hand 516:519 the other hand 506:519 the other hand 506:519 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 4 73 theme polyvalent 1064:1073 arg1 antibodies 1079:1088 pooled polyvalent IgG antibodies 1057:1088 pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 1057:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 3 74 dep require 635:641 arg1 afford 753:758 afford 753:758 afford 753:758 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 3 75 theme capabilities 685:696 arg1 advantage 668:676 advantage 668:676 advantage of the capabilities of automated microarray readers 668:728 On the other hand, lectins as probes on microarrays have recently been used in orthogonal approaches for in situ glycoprofiling but require analyte labeling to take advantage of the capabilities of automated microarray readers and data analysis they afford. 25851037 4 76 theme lectin-based 783:794 arg1 [ELISA 855:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 lectin-enzyme-linked immunosorbent assay [ELISA 814:860 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 4 76 theme lectin-based 783:794 arg1 assay 807:811 a lectin-based microtiter assay 781:811 a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors 781:1136 Herein, we describe a lectin-based microtiter assay (lectin-enzyme-linked immunosorbent assay [ELISA]) to quantify terminal glycan moieties, applicable to in vitro and in-cell glycan-engineered Fc proteins as well as intact IgGs from intravenous immunoglobulin (IVIG), a blood product containing pooled polyvalent IgG antibodies extracted from plasma from healthy human donors. 25851037 6 77 theme " 1225:1225 arg1 ELISA 1227:1231 This "customizable" ELISA 1207:1231 This "customizable" ELISA 1207:1231 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 25851037 6 78 theme dual-arm 1363:1370 arg1 glycosylation 1372:1384 single- versus dual-arm glycosylation 1348:1384 single- versus dual-arm glycosylation 1348:1384 This "customizable" ELISA juxtaposes readouts from multiple lectins, focusing on a subset of glycoforms, and provides the ability to discern single- versus dual-arm glycosylation while defining levels of epitopes at sensitivities comparable to MS. Extendable to other biologics, this ELISA can be used stand-alone or complementary to MS for quantitative glycan analysis. 28754360 5 0 theme production 1176:1185 arg1 rate 1187:1190 the IgG production rate 1168:1190 the IgG production rate 1168:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 4 1 theme constraint-based 702:717 arg1 modeling 719:726 a constraint-based modeling 700:726 a constraint-based modeling of the glycosylation network 700:755 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 8 2 theme global 1754:1759 arg1 factors 1790:1796 local (enzyme-specific) and global (cell specific productivity) factors 1726:1796 local (enzyme-specific) and global (cell specific productivity) factors 1726:1796 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 6 3 theme CHO 1208:1210 arg1 cultivations 1222:1233 the CHO fed-batch cultivations 1204:1233 the CHO fed-batch cultivations 1204:1233 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 0 4 theme distribution 88:99 arg1 changes 44:50 dynamic changes 36:50 dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures 36:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 8 5 theme specific 1767:1774 arg1 productivity 1776:1787 cell specific productivity 1762:1787 cell specific productivity 1762:1787 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 8 6 dep local 1726:1730 arg1 productivity 1776:1787 cell specific productivity 1762:1787 cell specific productivity 1762:1787 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 8 6 dep local 1726:1730 arg1 enzyme-specific 1733:1747 enzyme-specific 1733:1747 enzyme-specific 1733:1747 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 0 7 theme hamster 112:118 arg1 ovary 120:124 Chinese hamster ovary 104:124 Chinese hamster ovary fed-batch cultures 104:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 5 8 theme enzyme-specific 963:977 arg1 factor 979:984 an enzyme-specific factor 960:984 (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme 956:1078 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 8 9 theme dynamic 1675:1681 arg1 alterations 1683:1693 the dynamic alterations 1671:1693 the dynamic alterations of the glycosylation fluxes 1671:1721 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 5 10 theme temporal 1004:1011 arg1 changes 1013:1019 the temporal changes 1000:1019 the temporal changes among glycosylation reactions catalysed by the same enzyme 1000:1078 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 5 11 theme factors 947:953 arg1 way 928:930 way 928:930 way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate 928:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 5 12 theme IgG 1172:1174 arg1 rate 1187:1190 the IgG production rate 1168:1190 the IgG production rate 1168:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 0 13 theme Chinese 104:110 arg1 ovary 120:124 Chinese hamster ovary 104:124 Chinese hamster ovary fed-batch cultures 104:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 7 14 theme period 1597:1602 arg1 beginning 1568:1576 the beginning 1564:1576 the beginning of the cultivation period 1564:1602 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 6 15 theme cultivations 1222:1233 arg1 GFA 1197:1199 The GFA 1193:1199 The GFA of the CHO fed-batch cultivations 1193:1233 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 4 16 theme state 784:788 arg1 assumption 790:799 a pseudo steady state assumption 768:799 a pseudo steady state assumption 768:799 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 4 17 theme network 749:755 arg1 modeling 719:726 a constraint-based modeling 700:726 a constraint-based modeling of the glycosylation network 700:755 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 0 18 theme fed-batch 126:134 arg1 cultures 136:143 Chinese hamster ovary fed-batch cultures 104:143 Chinese hamster ovary fed-batch cultures 104:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 1 19 contain has 181:183 arg1 glycosylation 155:167 N-linked glycosylation 146:167 N-linked glycosylation of proteins 146:179 N-linked glycosylation of proteins has both functional and structural significance. 28754360 1 19 contain has 181:183 arg2 significance 216:227 both functional and structural significance 185:227 both functional and structural significance 185:227 N-linked glycosylation of proteins has both functional and structural significance. 28754360 6 20 theme fed-batch 1212:1220 arg1 cultivations 1222:1233 the CHO fed-batch cultivations 1204:1233 the CHO fed-batch cultivations 1204:1233 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 7 21 contain has 1492:1494 arg1 addition 1439:1446 the addition 1435:1446 the addition of Mn, a cofactor of galactosyltransferase, 1435:1490 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 7 21 contain has 1492:1494 arg2 effect 1500:1505 the effect 1496:1505 the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period 1496:1602 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 3 22 theme immunoglobulin 559:572 arg1 IgG 577:579 IgG 577:579 IgG 577:579 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 22 theme immunoglobulin 559:572 arg1 G 574:574 CHO fed-batch immunoglobulin G 545:574 CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding 545:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 1 23 theme functional 190:199 arg1 significance 216:227 both functional and structural significance 185:227 both functional and structural significance 185:227 N-linked glycosylation of proteins has both functional and structural significance. 28754360 0 24 theme ovary 120:124 arg1 cultures 136:143 Chinese hamster ovary fed-batch cultures 104:143 Chinese hamster ovary fed-batch cultures 104:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 0 25 from changes 44:50 arg1 cultures 136:143 Chinese hamster ovary fed-batch cultures 104:143 Chinese hamster ovary fed-batch cultures 104:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 5 26 from changes 1157:1163 arg1 rate 1187:1190 the IgG production rate 1168:1190 the IgG production rate 1168:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 7 27 theme galactosylation 1525:1539 arg1 fluxes 1541:1546 the galactosylation fluxes 1521:1546 the galactosylation fluxes 1521:1546 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 3 28 theme G 574:574 arg1 production 582:591 CHO fed-batch immunoglobulin G (IgG) production 545:591 CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding 545:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 0 29 theme flux 14:17 arg1 analysis 19:26 Glycosylation flux analysis 0:26 Glycosylation flux analysis 0:26 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 5 30 dep factor 1120:1125 arg1 2 1086:1086 2 1086:1086 2 1086:1086 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 6 31 theme media 1265:1269 arg1 composition 1271:1281 the media composition 1261:1281 the media composition 1261:1281 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 1 32 theme structural 205:214 arg1 significance 216:227 both functional and structural significance 185:227 both functional and structural significance 185:227 N-linked glycosylation of proteins has both functional and structural significance. 28754360 0 33 theme Glycosylation 0:12 arg1 analysis 19:26 Glycosylation flux analysis 0:26 Glycosylation flux analysis 0:26 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 3 34 theme consumption 474:484 arg1 rates 486:490 consumption rates 474:490 consumption rates (fluxes) 474:499 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 34 theme consumption 474:484 arg1 fluxes 493:498 fluxes 493:498 fluxes 493:498 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 5 35 theme factor 979:984 arg1 way 928:930 way 928:930 way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate 928:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 3 36 theme media 613:617 arg1 compositions 619:630 two different media compositions 599:630 two different media compositions 599:630 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 4 37 theme steady 777:782 arg1 assumption 790:799 a pseudo steady state assumption 768:799 a pseudo steady state assumption 768:799 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 5 38 dep factor 979:984 arg1 1 957:957 1 957:957 1 957:957 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 5 39 theme dynamic 1149:1155 arg1 changes 1157:1163 the dynamic changes 1145:1163 the dynamic changes in the IgG production rate 1145:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 6 40 theme other 1371:1375 arg1 reactions 1391:1399 the other glycosylation reactions 1367:1399 the other glycosylation reactions 1367:1399 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 4 41 theme pseudo 770:775 arg1 assumption 790:799 a pseudo steady state assumption 768:799 a pseudo steady state assumption 768:799 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 8 42 theme fluxes 1716:1721 arg1 alterations 1683:1693 the dynamic alterations 1671:1693 the dynamic alterations of the glycosylation fluxes 1671:1721 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 3 43 theme glycoforms 504:513 arg1 rates 486:490 consumption rates 474:490 consumption rates (fluxes) 474:499 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 43 theme glycoforms 504:513 arg1 fluxes 493:498 fluxes 493:498 fluxes 493:498 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 43 theme glycoforms 504:513 arg1 production 459:468 intracellular production 445:468 intracellular production 445:468 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 44 theme intracellular 445:457 arg1 production 459:468 intracellular production 445:468 intracellular production 445:468 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 1 45 gly glycosylation 155:167 arg1 proteins 172:179 proteins 172:179 proteins 172:179 N-linked glycosylation of proteins has both functional and structural significance. 28754360 8 46 theme glycosylation 1702:1714 arg1 fluxes 1716:1721 the glycosylation fluxes 1698:1721 the glycosylation fluxes 1698:1721 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 5 47 theme time 869:872 arg1 point 874:878 each time point 864:878 each time point 864:878 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 0 48 theme dynamic 36:42 arg1 changes 44:50 dynamic changes 36:50 dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures 36:143 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 5 49 theme productivity 1107:1118 arg1 factor 1120:1125 the cell specific productivity factor 1089:1125 (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate 1085:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 5 50 theme glycosylation 812:824 arg1 fluxes 826:831 the glycosylation fluxes 808:831 the glycosylation fluxes in the network 808:846 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 3 51 theme manganese 661:669 arg1 feeding 671:677 additional manganese feeding 650:677 additional manganese feeding 650:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 8 52 theme local 1726:1730 arg1 factors 1790:1796 local (enzyme-specific) and global (cell specific productivity) factors 1726:1796 local (enzyme-specific) and global (cell specific productivity) factors 1726:1796 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 0 53 theme intracellular 55:67 arg1 distribution 88:99 intracellular glycosylation flux distribution 55:99 intracellular glycosylation flux distribution 55:99 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 3 54 theme glycosylation 396:408 arg1 GFA 425:427 GFA 425:427 GFA 425:427 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 54 theme glycosylation 396:408 arg1 analysis 415:422 glycosylation flux analysis 396:422 glycosylation flux analysis (GFA) 396:428 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 1 55 link N-linked 146:153 arg1 glycosylation 155:167 N-linked glycosylation 146:167 N-linked glycosylation of proteins 146:179 N-linked glycosylation of proteins has both functional and structural significance. 28754360 6 56 theme galactosylation 1284:1298 arg1 fluxes 1300:1305 galactosylation fluxes 1284:1305 galactosylation fluxes 1284:1305 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 3 57 theme different 603:611 arg1 compositions 619:630 two different media compositions 599:630 two different media compositions 599:630 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 5 58 from point 874:878 arg1 balanced 852:859 balanced 852:859 balanced 852:859 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 4 59 theme glycosylation 735:747 arg1 network 749:755 the glycosylation network 731:755 the glycosylation network 731:755 The GFA is based on a constraint-based modeling of the glycosylation network, employing a pseudo steady state assumption. 28754360 5 60 theme specific 1098:1105 arg1 factor 1120:1125 the cell specific productivity factor 1089:1125 (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate 1085:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 6 61 theme glycosylation 1377:1389 arg1 reactions 1391:1399 the other glycosylation reactions 1367:1399 the other glycosylation reactions 1367:1399 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 5 62 from fluxes 826:831 arg1 network 840:846 the network 836:846 the network 836:846 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 7 63 theme galactosyltransferase 1469:1489 arg1 Mn 1451:1452 Mn 1451:1452 Mn 1451:1452 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 7 63 theme galactosyltransferase 1469:1489 arg1 cofactor 1457:1464 a cofactor 1455:1464 a cofactor of galactosyltransferase 1455:1489 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 2 64 theme glycan 247:252 arg1 structure 254:262 the glycan structure 243:262 the glycan structure of a therapeutic protein 243:287 Importantly, the glycan structure of a therapeutic protein influences its efficacy, pharmacokinetics, pharmacodynamics and immunogenicity. 28754360 5 65 theme same 1068:1071 arg1 enzyme 1073:1078 the same enzyme 1064:1078 the same enzyme 1064:1078 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 7 66 theme Mn 1451:1452 arg1 addition 1439:1446 the addition 1435:1446 the addition of Mn, a cofactor of galactosyltransferase, 1435:1490 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 5 67 theme glycosylation 1027:1039 arg1 reactions 1041:1049 glycosylation reactions 1027:1049 glycosylation reactions catalysed by the same enzyme 1027:1078 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 8 68 theme cell 1762:1765 arg1 productivity 1776:1787 cell specific productivity 1762:1787 cell specific productivity 1762:1787 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 1 69 theme N-linked 146:153 arg1 glycosylation 155:167 N-linked glycosylation 146:167 N-linked glycosylation of proteins 146:179 N-linked glycosylation of proteins has both functional and structural significance. 28754360 5 70 theme scaling 939:945 arg1 factors 947:953 two scaling factors 935:953 two scaling factors 935:953 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 3 71 theme additional 650:659 arg1 feeding 671:677 additional manganese feeding 650:677 additional manganese feeding 650:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 7 72 theme cultivation 1585:1595 arg1 period 1597:1602 the cultivation period 1581:1602 the cultivation period 1581:1602 Furthermore, the GFA showed that the addition of Mn, a cofactor of galactosyltransferase, has the effect of increasing the galactosylation fluxes but only during the beginning of the cultivation period. 28754360 3 73 theme fed-batch 549:557 arg1 IgG 577:579 IgG 577:579 IgG 577:579 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 73 theme fed-batch 549:557 arg1 G 574:574 CHO fed-batch immunoglobulin G 545:574 CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding 545:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 6 74 theme cultivation 1326:1336 arg1 time 1338:1341 the cultivation time 1322:1341 the cultivation time 1322:1341 The GFA of the CHO fed-batch cultivations showed that regardless of the media composition, galactosylation fluxes decreased with the cultivation time more significantly than the other glycosylation reactions. 28754360 0 75 theme flux 83:86 arg1 distribution 88:99 intracellular glycosylation flux distribution 55:99 intracellular glycosylation flux distribution 55:99 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 8 76 theme GFA 1652:1654 arg1 power 1639:1643 the power 1635:1643 the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors 1635:1796 The results thus demonstrated the power of the GFA in delineating the dynamic alterations of the glycosylation fluxes by local (enzyme-specific) and global (cell specific productivity) factors. 28754360 2 77 theme protein 281:287 arg1 structure 254:262 the glycan structure 243:262 the glycan structure of a therapeutic protein 243:287 Importantly, the glycan structure of a therapeutic protein influences its efficacy, pharmacokinetics, pharmacodynamics and immunogenicity. 28754360 3 78 theme CHO 545:547 arg1 IgG 577:579 IgG 577:579 IgG 577:579 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 78 theme CHO 545:547 arg1 G 574:574 CHO fed-batch immunoglobulin G 545:574 CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding 545:677 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 0 79 theme glycosylation 69:81 arg1 distribution 88:99 intracellular glycosylation flux distribution 55:99 intracellular glycosylation flux distribution 55:99 Glycosylation flux analysis reveals dynamic changes of intracellular glycosylation flux distribution in Chinese hamster ovary fed-batch cultures. 28754360 5 80 theme factor 1120:1125 arg1 way 928:930 way 928:930 way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate 928:1190 While the glycosylation fluxes in the network are balanced at each time point, the GFA allows the fluxes to vary with time by way of two scaling factors: (1) an enzyme-specific factor that captures the temporal changes among glycosylation reactions catalysed by the same enzyme, and (2) the cell specific productivity factor that accounts for the dynamic changes in the IgG production rate. 28754360 2 81 theme therapeutic 269:279 arg1 protein 281:287 a therapeutic protein 267:287 a therapeutic protein 267:287 Importantly, the glycan structure of a therapeutic protein influences its efficacy, pharmacokinetics, pharmacodynamics and immunogenicity. 28754360 1 82 theme proteins 172:179 arg1 glycosylation 155:167 N-linked glycosylation 146:167 N-linked glycosylation of proteins 146:179 N-linked glycosylation of proteins has both functional and structural significance. 28754360 3 83 theme flux 410:413 arg1 GFA 425:427 GFA 425:427 GFA 425:427 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 28754360 3 83 theme flux 410:413 arg1 analysis 415:422 glycosylation flux analysis 396:422 glycosylation flux analysis (GFA) 396:428 In this work, we developed glycosylation flux analysis (GFA) for predicting intracellular production and consumption rates (fluxes) of glycoforms, and applied this analysis to CHO fed-batch immunoglobulin G (IgG) production using two different media compositions, with and without additional manganese feeding. 24575722 6 0 theme fucosylated 1150:1160 arg1 haptoglobin 1162:1172 fucosylated haptoglobin 1150:1172 fucosylated haptoglobin 1150:1172 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 6 0 theme fucosylated 1150:1160 arg1 AUC 1185:1187 AUC = 0.88 1185:1194 AUC = 0.88 1185:1194 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 4 1 attach removed 754:760 arg2 acid 749:752 sialic acid 742:752 sialic acid removed from the glycoproteins 742:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 4 1 attach removed 754:760 arg1 glycoproteins 771:783 the glycoproteins 767:783 the glycoproteins 767:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 6 2 theme stage-III 1244:1252 arg1 cancer 1262:1267 stage-III ovarian cancer 1244:1267 stage-III ovarian cancer 1244:1267 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 4 3 theme abundance 891:899 arg1 variation 901:909 protein abundance variation 883:909 glycosylation changes of proteins rather than protein abundance variation 837:909 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 6 4 theme = 1314:1314 arg1 0.86 1316:1319 AUC = 0.86 1310:1319 AUC = 0.86 1310:1319 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 6 4 theme = 1314:1314 arg1 CA125 1297:1301 CA125 1297:1301 CA125 alone (AUC = 0.86) 1297:1320 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 2 5 theme diagnostic 288:297 arg1 markers 309:315 potential diagnostic candidate markers 278:315 potential diagnostic candidate markers 278:315 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 1 6 theme certain 176:182 arg1 glycoproteins 206:218 glycoproteins 206:218 glycoproteins 206:218 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 6 theme certain 176:182 arg1 markers 235:241 well-known markers 224:241 well-known markers for tumor progression 224:263 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 6 theme certain 176:182 arg1 structures 191:200 certain glycan structures 176:200 certain glycan structures 176:200 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 6 7 theme improved 1204:1211 arg1 performance 1213:1223 improved performance 1204:1223 improved performance for distinguishing stage-III ovarian cancer from noncases 1204:1281 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 5 8 with patients 1014:1021 arg1 cancer 1036:1041 ovarian cancer 1028:1041 ovarian cancer 1028:1041 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 5 9 theme lectin-based 930:941 arg1 assay 949:953 our reverse lectin-based ELISA assay 918:953 our reverse lectin-based ELISA assay 918:953 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 2 10 from changes 381:387 arg1 samples 423:429 crude serum samples 411:429 crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 411:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 1 11 gly glycosylation 106:118 arg1 glycoproteins 123:135 glycoproteins 123:135 glycoproteins 123:135 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 3 12 theme low 608:610 arg1 background 612:621 low background 608:621 low background 608:621 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 1 13 gly glycoproteins 123:135 arg1 glycoproteins 123:135 glycoproteins 123:135 glycoproteins 123:135 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 14 theme well-known 224:233 arg1 glycoproteins 206:218 glycoproteins 206:218 glycoproteins 206:218 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 14 theme well-known 224:233 arg1 markers 235:241 well-known markers 224:241 well-known markers for tumor progression 224:263 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 14 theme well-known 224:233 arg1 structures 191:200 certain glycan structures 176:200 certain glycan structures 176:200 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 8 15 theme 0.855 1523:1527 arg1 AUC 1516:1518 an AUC 1513:1518 an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases 1513:1600 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 24575722 2 16 with detection 483:491 arg1 antibodies 520:529 biotin/HRP-conjugated antibodies 498:529 biotin/HRP-conjugated antibodies 498:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 4 17 theme AAL 730:732 arg1 sugars 713:718 sugars 713:718 sugars of lectin AAL or with sialic acid removed from the glycoproteins 713:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 3 18 theme lectin 662:667 arg1 methods 675:681 current lectin ELISA methods 654:681 current lectin ELISA methods 654:681 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 0 19 theme reverse 64:70 arg1 assay 91:95 the reverse lectin-based ELISA assay 60:95 the reverse lectin-based ELISA assay 60:95 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 5 20 located observed 994:1001 arg2 haptoglobin 978:988 increased fucosylated haptoglobin 956:988 increased fucosylated haptoglobin 956:988 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 5 20 located observed 994:1001 arg1 sera 1006:1009 sera 1006:1009 sera of patients with ovarian cancer 1006:1041 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 5 21 theme increased 956:964 arg1 haptoglobin 978:988 increased fucosylated haptoglobin 956:988 increased fucosylated haptoglobin 956:988 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 0 22 from Analysis 0:7 arg1 glycoproteins 32:44 glycoproteins 32:44 glycoproteins from serum 32:55 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 1 23 theme Altered 98:104 arg1 glycosylation 106:118 Altered glycosylation 98:118 Altered glycosylation in glycoproteins 98:135 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 0 24 theme ELISA 85:89 arg1 assay 91:95 the reverse lectin-based ELISA assay 60:95 the reverse lectin-based ELISA assay 60:95 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 5 25 theme ovarian 1028:1034 arg1 cancer 1036:1041 ovarian cancer 1028:1041 ovarian cancer 1028:1041 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 2 26 theme serum 417:421 arg1 samples 423:429 crude serum samples 411:429 crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 411:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 7 27 gly fucosylated 1384:1394 arg1 haptoglobin 1396:1406 fucosylated haptoglobin 1384:1406 fucosylated haptoglobin 1384:1406 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 9 28 from samples 1701:1707 arg1 proteins 1681:1688 proteins 1681:1688 proteins from serum samples, which will be essential for biomarker discovery and validation studies 1681:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 9 28 from samples 1701:1707 arg1 changes 1670:1676 glycosylation changes 1656:1676 glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies 1656:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 2 29 gly glycoproteins 392:404 arg1 glycoproteins 392:404 glycoproteins 392:404 glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 392:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 1 30 from glycosylation 106:118 arg1 glycoproteins 123:135 glycoproteins 123:135 glycoproteins 123:135 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 4 31 theme proteins 862:869 arg1 changes 851:857 glycosylation changes 837:857 glycosylation changes of proteins rather than protein abundance variation 837:909 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 2 32 theme glycoprotein 450:461 arg1 capture 463:469 lectin-based glycoprotein capture 437:469 lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 437:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 0 33 theme glycan 12:17 arg1 variation 19:27 glycan variation 12:27 glycan variation on glycoproteins from serum 12:55 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 5 34 theme patients 1014:1021 arg1 sera 1006:1009 sera 1006:1009 sera of patients with ovarian cancer 1006:1041 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 4 35 with sugars 713:718 arg1 acid 749:752 sialic acid 742:752 sialic acid removed from the glycoproteins 742:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 8 36 theme CA125 1467:1471 arg1 combination 1452:1462 The combination 1448:1462 The combination of CA125 and fucosylated haptoglobin 1448:1499 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 24575722 5 37 theme haptoglobin 1071:1081 arg1 level 1062:1066 the protein level 1050:1066 the protein level of haptoglobin 1050:1081 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 7 38 theme ovarian 1354:1360 arg1 cancer 1362:1367 early-stage ovarian cancer 1342:1367 early-stage ovarian cancer from noncases 1342:1381 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 2 39 theme biotin/HRP-conjugated 498:518 arg1 antibodies 520:529 biotin/HRP-conjugated antibodies 498:529 biotin/HRP-conjugated antibodies 498:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 8 40 theme fucosylated 1477:1487 arg1 haptoglobin 1489:1499 fucosylated haptoglobin 1477:1499 fucosylated haptoglobin 1477:1499 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 24575722 8 41 theme early-stage 1569:1579 arg1 cancer 1581:1586 early-stage cancer 1569:1586 early-stage cancer 1569:1586 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 24575722 9 42 theme serum 1695:1699 arg1 samples 1701:1707 serum samples 1695:1707 serum samples 1695:1707 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 6 43 theme ovarian 1254:1260 arg1 cancer 1262:1267 stage-III ovarian cancer 1244:1267 stage-III ovarian cancer 1244:1267 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 9 44 theme proteins 1681:1688 arg1 changes 1670:1676 glycosylation changes 1656:1676 glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies 1656:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 0 45 gly glycoproteins 32:44 arg1 glycoproteins 32:44 glycoproteins 32:44 glycoproteins from serum 32:55 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 7 46 from noncases 1374:1381 arg1 cancer 1362:1367 early-stage ovarian cancer 1342:1367 early-stage ovarian cancer from noncases 1342:1381 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 6 47 theme CA125 1178:1182 arg1 combination 1135:1145 The combination 1131:1145 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) 1131:1195 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 1 48 theme tumor 247:251 arg1 progression 253:263 tumor progression 247:263 tumor progression 247:263 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 6 49 theme haptoglobin 1162:1172 arg1 combination 1135:1145 The combination 1131:1145 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) 1131:1195 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 2 50 theme glycosylation 367:379 arg1 changes 381:387 glycosylation changes 367:387 glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 367:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 9 51 theme biomarker 1738:1746 arg1 discovery 1748:1756 biomarker discovery 1738:1756 biomarker discovery 1738:1756 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 1 52 gly glycoproteins 206:218 arg1 glycoproteins 206:218 glycoproteins 206:218 glycoproteins 206:218 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 52 gly glycoproteins 206:218 arg1 markers 235:241 well-known markers 224:241 well-known markers for tumor progression 224:263 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 52 gly glycoproteins 206:218 arg1 structures 191:200 certain glycan structures 176:200 certain glycan structures 176:200 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 2 53 theme candidate 299:307 arg1 markers 309:315 potential diagnostic candidate markers 278:315 potential diagnostic candidate markers 278:315 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 1 54 theme glycan 184:189 arg1 glycoproteins 206:218 glycoproteins 206:218 glycoproteins 206:218 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 54 theme glycan 184:189 arg1 markers 235:241 well-known markers 224:241 well-known markers for tumor progression 224:263 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 1 54 theme glycan 184:189 arg1 structures 191:200 certain glycan structures 176:200 certain glycan structures 176:200 Altered glycosylation in glycoproteins is associated with carcinogenesis, and certain glycan structures and glycoproteins are well-known markers for tumor progression. 24575722 4 55 theme protein 883:889 arg1 variation 901:909 protein abundance variation 883:909 glycosylation changes of proteins rather than protein abundance variation 837:909 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 6 56 theme AUC 1310:1312 arg1 0.86 1316:1319 AUC = 0.86 1310:1319 AUC = 0.86 1310:1319 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 6 56 theme AUC 1310:1312 arg1 CA125 1297:1301 CA125 1297:1301 CA125 alone (AUC = 0.86) 1297:1320 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 5 57 theme reverse 922:928 arg1 assay 949:953 our reverse lectin-based ELISA assay 918:953 our reverse lectin-based ELISA assay 918:953 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 5 58 gly fucosylated 966:976 arg1 haptoglobin 978:988 increased fucosylated haptoglobin 956:988 increased fucosylated haptoglobin 956:988 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 3 59 theme improved 627:634 arg1 S/N 636:638 improved S/N 627:638 improved S/N 627:638 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 5 60 theme fucosylated 966:976 arg1 haptoglobin 978:988 increased fucosylated haptoglobin 956:988 increased fucosylated haptoglobin 956:988 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 4 61 theme sialic 742:747 arg1 acid 749:752 sialic acid 742:752 sialic acid removed from the glycoproteins 742:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 3 62 theme ELISA 669:673 arg1 methods 675:681 current lectin ELISA methods 654:681 current lectin ELISA methods 654:681 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 9 63 from changes 1670:1676 arg1 samples 1701:1707 serum samples 1695:1707 serum samples 1695:1707 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 3 64 theme current 654:660 arg1 methods 675:681 current lectin ELISA methods 654:681 current lectin ELISA methods 654:681 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 5 65 theme ELISA 943:947 arg1 assay 949:953 our reverse lectin-based ELISA assay 918:953 our reverse lectin-based ELISA assay 918:953 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 0 66 from variation 19:27 arg1 glycoproteins 32:44 glycoproteins 32:44 glycoproteins from serum 32:55 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 7 67 theme comparable 1415:1424 arg1 performance 1426:1436 comparable performance 1415:1436 comparable performance 1415:1436 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 4 68 theme lectin 723:728 arg1 AAL 730:732 lectin AAL 723:732 lectin AAL 723:732 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 4 69 gly glycoproteins 771:783 arg1 glycoproteins 771:783 the glycoproteins 767:783 the glycoproteins 767:783 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 0 70 theme lectin-based 72:83 arg1 assay 91:95 the reverse lectin-based ELISA assay 60:95 the reverse lectin-based ELISA assay 60:95 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 2 71 gly glycoprotein 450:461 arg1 glycoprotein 450:461 lectin-based glycoprotein capture 437:469 lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 437:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 0 72 from serum 51:55 arg1 glycoproteins 32:44 glycoproteins 32:44 glycoproteins from serum 32:55 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 6 73 gly fucosylated 1150:1160 arg1 haptoglobin 1162:1172 fucosylated haptoglobin 1150:1172 fucosylated haptoglobin 1150:1172 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 6 73 gly fucosylated 1150:1160 arg1 AUC 1185:1187 AUC = 0.88 1185:1194 AUC = 0.88 1185:1194 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 9 74 theme alternative 1625:1635 arg1 method 1637:1642 an alternative method 1622:1642 an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies 1622:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 7 75 theme fucosylated 1384:1394 arg1 haptoglobin 1396:1406 fucosylated haptoglobin 1384:1406 fucosylated haptoglobin 1384:1406 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 0 76 from glycoproteins 32:44 arg1 Analysis 0:7 Analysis 0:7 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 0:96 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 2 77 theme crude 411:415 arg1 samples 423:429 crude serum samples 411:429 crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 411:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 2 78 theme glycoproteins 392:404 arg1 changes 381:387 glycosylation changes 367:387 glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 367:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 0 79 theme variation 19:27 arg1 Analysis 0:7 Analysis 0:7 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 0:96 Analysis of glycan variation on glycoproteins from serum by the reverse lectin-based ELISA assay. 24575722 4 80 dep competing 703:711 arg1 the 687:689 the 687:689 the 687:689 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 4 80 dep competing 703:711 arg1 presence 691:698 presence 691:698 presence 691:698 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 9 81 theme glycosylation 1656:1668 arg1 changes 1670:1676 glycosylation changes 1656:1676 glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies 1656:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 2 82 theme lectin-based 437:448 arg1 capture 463:469 lectin-based glycoprotein capture 437:469 lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 437:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 7 83 theme early-stage 1342:1352 arg1 cancer 1362:1367 early-stage ovarian cancer 1342:1367 early-stage ovarian cancer from noncases 1342:1381 In differentiating early-stage ovarian cancer from noncases, fucosylated haptoglobin showed comparable performance to CA125. 24575722 9 84 dep discovery 1748:1756 arg1 studies 1773:1779 studies 1773:1779 studies 1773:1779 Our study provides an alternative method to quantify glycosylation changes of proteins from serum samples, which will be essential for biomarker discovery and validation studies. 24575722 6 85 theme = 1189:1189 arg1 haptoglobin 1162:1172 fucosylated haptoglobin 1150:1172 fucosylated haptoglobin 1150:1172 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 6 85 theme = 1189:1189 arg1 AUC 1185:1187 AUC = 0.88 1185:1194 AUC = 0.88 1185:1194 The combination of fucosylated haptoglobin and CA125 (AUC = 0.88) showed improved performance for distinguishing stage-III ovarian cancer from noncases compared with CA125 alone (AUC = 0.86). 24575722 2 86 theme changes 381:387 arg1 analysis 355:362 analysis 355:362 analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 355:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 3 87 theme lectin 546:551 arg1 lectin 546:551 lectin 546:551 lectin 546:551 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 3 87 theme lectin 546:551 arg1 amount 536:541 The amount 532:541 The amount of lectin coated on the microplate well 532:581 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 8 88 theme haptoglobin 1489:1499 arg1 combination 1452:1462 The combination 1448:1462 The combination of CA125 and fucosylated haptoglobin 1448:1499 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 24575722 2 89 theme novel 338:342 arg1 method 344:349 a novel method 336:349 a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 336:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 5 90 theme protein 1054:1060 arg1 level 1062:1066 the protein level 1050:1066 the protein level of haptoglobin 1050:1081 Using our reverse lectin-based ELISA assay, increased fucosylated haptoglobin was observed in sera of patients with ovarian cancer, while the protein level of haptoglobin remained the same between cancers and noncases. 24575722 2 91 from samples 423:429 arg1 glycoproteins 392:404 glycoproteins 392:404 glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 392:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 2 91 from samples 423:429 arg1 changes 381:387 glycosylation changes 367:387 glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies 367:529 To identify potential diagnostic candidate markers, we have developed a novel method for analysis of glycosylation changes of glycoproteins from crude serum samples using lectin-based glycoprotein capture followed by detection with biotin/HRP-conjugated antibodies. 24575722 4 92 theme glycosylation 837:849 arg1 changes 851:857 glycosylation changes 837:857 glycosylation changes of proteins rather than protein abundance variation 837:909 In the presence of competing sugars of lectin AAL or with sialic acid removed from the glycoproteins, we confirmed that this method specifically detects glycosylation changes of proteins rather than protein abundance variation. 24575722 3 93 theme microplate 567:576 arg1 well 578:581 the microplate well 563:581 the microplate well 563:581 The amount of lectin coated on the microplate well was optimized to achieve low background and improved S/N compared with current lectin ELISA methods. 24575722 8 94 gly fucosylated 1477:1487 arg1 haptoglobin 1489:1499 fucosylated haptoglobin 1477:1499 fucosylated haptoglobin 1477:1499 The combination of CA125 and fucosylated haptoglobin resulted in an AUC of 0.855, which outperforms CA125 to distinguish early-stage cancer from noncases. 27485625 4 0 link N-linked 696:703 arg1 glycans 705:711 the N-linked glycans 692:711 the N-linked glycans from standard glycoprotein or complex human serum proteins 692:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 5 1 theme enrichment 858:867 arg1 efficiency 869:878 good enrichment efficiency 853:878 The simple synthesis method as well as good enrichment efficiency 814:878 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 1 2 theme key 176:178 arg1 role 180:183 a key role 174:183 a key role 174:183 Protein glycosylation plays a key role in many biological processes. 27485625 0 3 theme selective 104:112 arg1 enrichment 114:123 highly selective enrichment 97:123 highly selective enrichment of N-linked glycans 97:143 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 5 4 theme good 853:856 arg1 efficiency 869:878 good enrichment efficiency 853:878 The simple synthesis method as well as good enrichment efficiency 814:878 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 4 5 theme standard 718:725 arg1 glycoprotein 727:738 standard glycoprotein 718:738 standard glycoprotein 718:738 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 4 6 from glycoprotein 727:738 arg1 glycans 705:711 the N-linked glycans 692:711 the N-linked glycans from standard glycoprotein or complex human serum proteins 692:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 3 7 with ions 381:384 arg1 linkers 408:414 bridging organic linkers 391:414 bridging organic linkers 391:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 1 8 theme Protein 146:152 arg1 glycosylation 154:166 Protein glycosylation 146:166 Protein glycosylation 146:166 Protein glycosylation plays a key role in many biological processes. 27485625 4 9 theme complex 743:749 arg1 proteins 763:770 complex human serum proteins 743:770 complex human serum proteins 743:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 3 10 theme organic 400:406 arg1 linkers 408:414 bridging organic linkers 391:414 bridging organic linkers 391:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 5 11 theme synthesis 825:833 arg1 method 835:840 The simple synthesis method 814:840 The simple synthesis method as well as good enrichment efficiency 814:878 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 3 12 theme many 420:423 arg1 amount 533:538 especially a large amount 514:538 especially a large amount of carbon 514:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 12 theme many 420:423 arg1 properties 437:446 many fascinating properties 420:446 many fascinating properties 420:446 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 12 theme many 420:423 arg1 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 12 theme many 420:423 arg1 carbon 543:548 carbon 543:548 carbon 543:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 12 theme many 420:423 arg1 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 0 13 theme novel 2:6 arg1 material 15:22 A novel carbon material 0:22 A novel carbon material with nanopores prepared 0:46 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 4 14 theme N-linked 696:703 arg1 glycans 705:711 the N-linked glycans 692:711 the N-linked glycans from standard glycoprotein or complex human serum proteins 692:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 0 15 link N-linked 128:135 arg1 glycans 137:143 N-linked glycans 128:143 N-linked glycans 128:143 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 1 16 theme many 188:191 arg1 processes 204:212 many biological processes 188:212 many biological processes 188:212 Protein glycosylation plays a key role in many biological processes. 27485625 2 17 theme framework 316:324 arg1 Mil-101 332:338 metal-organic framework (MOF) Mil-101 302:338 metal-organic framework (MOF) Mil-101(Cr) 302:342 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 2 17 theme framework 316:324 arg1 Cr 340:341 Cr 340:341 Cr 340:341 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 1 18 theme biological 193:202 arg1 processes 204:212 many biological processes 188:212 many biological processes 188:212 Protein glycosylation plays a key role in many biological processes. 27485625 3 19 theme parent 349:354 arg1 MOF 356:358 The parent MOF 345:358 The parent MOF assembled from metal ions with bridging organic linkers 345:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 0 20 theme glycans 137:143 arg1 enrichment 114:123 highly selective enrichment 97:123 highly selective enrichment of N-linked glycans 97:143 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 5 21 theme carbon 896:901 arg1 material 903:910 this novel carbon material 885:910 this novel carbon material 885:910 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 2 22 theme metal-organic 302:314 arg1 Mil-101 332:338 metal-organic framework (MOF) Mil-101 302:338 metal-organic framework (MOF) Mil-101(Cr) 302:342 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 2 22 theme metal-organic 302:314 arg1 Cr 340:341 Cr 340:341 Cr 340:341 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 0 23 theme carbon 8:13 arg1 material 15:22 A novel carbon material 0:22 A novel carbon material with nanopores prepared 0:46 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 0 24 theme N-linked 128:135 arg1 glycans 137:143 N-linked glycans 128:143 N-linked glycans 128:143 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 3 25 theme suitable 481:488 arg1 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 4 26 theme size-exclusion 645:658 arg1 effect 660:665 the size-exclusion effect 641:665 the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein 579:689 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 4 27 theme pore 670:673 arg1 effect 660:665 the size-exclusion effect 641:665 the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein 579:689 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 4 27 theme pore 670:673 arg1 interactions 590:601 the strong interactions 579:601 the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein 579:689 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 2 28 theme novel 232:236 arg1 material 245:252 a novel carbon material 230:252 a novel carbon material with nanopores 230:267 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 3 29 theme fascinating 425:435 arg1 amount 533:538 especially a large amount 514:538 especially a large amount of carbon 514:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 29 theme fascinating 425:435 arg1 properties 437:446 many fascinating properties 420:446 many fascinating properties 420:446 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 29 theme fascinating 425:435 arg1 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 29 theme fascinating 425:435 arg1 carbon 543:548 carbon 543:548 carbon 543:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 29 theme fascinating 425:435 arg1 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 30 theme bridging 391:398 arg1 linkers 408:414 bridging organic linkers 391:414 bridging organic linkers 391:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 2 31 with material 245:252 arg1 nanopores 259:267 nanopores 259:267 nanopores 259:267 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 0 32 with material 15:22 arg1 nanopores prepared 29:46 nanopores prepared 29:46 nanopores prepared 29:46 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 5 33 theme simple 818:823 arg1 method 835:840 The simple synthesis method 814:840 The simple synthesis method as well as good enrichment efficiency 814:878 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 0 34 theme metal-organic 56:68 arg1 framework 70:78 metal-organic framework 56:78 metal-organic framework 56:78 A novel carbon material with nanopores prepared using a metal-organic framework as precursor for highly selective enrichment of N-linked glycans. 27485625 4 35 gly glycoprotein 727:738 arg1 glycoprotein 727:738 standard glycoprotein 718:738 standard glycoprotein 718:738 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 4 36 theme high 797:800 arg1 efficiency 802:811 high efficiency 797:811 high efficiency 797:811 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 3 37 theme surface 467:473 arg1 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 5 38 theme promising 914:922 arg1 tool 924:927 a promising tool 912:927 a promising tool for glycosylation research 912:954 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 2 39 theme Mil-101 332:338 arg1 carbonization 285:297 carbonization 285:297 carbonization of metal-organic framework (MOF) Mil-101(Cr) 285:342 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 4 40 theme serum 757:761 arg1 proteins 763:770 complex human serum proteins 743:770 complex human serum proteins 743:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 5 41 theme novel 890:894 arg1 material 903:910 this novel carbon material 885:910 this novel carbon material 885:910 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 3 42 theme large 527:531 arg1 amount 533:538 especially a large amount 514:538 especially a large amount of carbon 514:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 42 theme large 527:531 arg1 carbon 543:548 carbon 543:548 carbon 543:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 4 43 theme human 751:755 arg1 proteins 763:770 complex human serum proteins 743:770 complex human serum proteins 743:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 2 44 theme MOF 327:329 arg1 Mil-101 332:338 metal-organic framework (MOF) Mil-101 302:338 metal-organic framework (MOF) Mil-101(Cr) 302:342 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 2 44 theme MOF 327:329 arg1 Cr 340:341 Cr 340:341 Cr 340:341 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 3 45 contain had 416:418 arg2 properties 437:446 many fascinating properties 420:446 many fascinating properties 420:446 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 45 contain had 416:418 arg2 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 45 contain had 416:418 arg2 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 45 contain had 416:418 arg2 amount 533:538 especially a large amount 514:538 especially a large amount of carbon 514:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 45 contain had 416:418 arg2 carbon 543:548 carbon 543:548 carbon 543:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 45 contain had 416:418 arg1 MOF 356:358 The parent MOF 345:358 The parent MOF assembled from metal ions with bridging organic linkers 345:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 4 46 from proteins 763:770 arg1 glycans 705:711 the N-linked glycans 692:711 the N-linked glycans from standard glycoprotein or complex human serum proteins 692:770 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 3 47 theme nanopore 490:497 arg1 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 48 theme ultrahigh 457:465 arg1 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 4 49 theme strong 583:588 arg1 interactions 590:601 the strong interactions 579:601 the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein 579:689 Due to the strong interactions between carbon and glycans as well as the size-exclusion effect of pore against protein, the N-linked glycans from standard glycoprotein or complex human serum proteins could be identified with high efficiency. 27485625 3 50 theme metal 375:379 arg1 ions 381:384 metal ions 375:384 metal ions with bridging organic linkers 375:414 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 5 51 theme glycosylation 933:945 arg1 research 947:954 glycosylation research 933:954 glycosylation research 933:954 The simple synthesis method as well as good enrichment efficiency made this novel carbon material a promising tool for glycosylation research. 27485625 2 52 theme carbon 238:243 arg1 material 245:252 a novel carbon material 230:252 a novel carbon material with nanopores 230:267 In this study, a novel carbon material with nanopores was prepared by carbonization of metal-organic framework (MOF) Mil-101(Cr). 27485625 3 53 theme carbon 543:548 arg1 amount 533:538 especially a large amount 514:538 especially a large amount of carbon 514:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 53 theme carbon 543:548 arg1 area 475:478 ultrahigh surface area 457:478 ultrahigh surface area 457:478 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 53 theme carbon 543:548 arg1 carbon 543:548 carbon 543:548 carbon 543:548 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 27485625 3 53 theme carbon 543:548 arg1 structure 499:507 suitable nanopore structure 481:507 suitable nanopore structure 481:507 The parent MOF assembled from metal ions with bridging organic linkers had many fascinating properties, such as ultrahigh surface area, suitable nanopore structure, and especially a large amount of carbon after being calcined. 24828102 0 0 theme antibody 97:104 arg1 drugs 106:110 manufactured recombinant monoclonal antibody drugs 61:110 manufactured recombinant monoclonal antibody drugs 61:110 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 3 1 theme N-glycan 509:516 arg1 library 518:524 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library 459:524 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs 459:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 0 2 theme monoclonal 86:95 arg1 drugs 106:110 manufactured recombinant monoclonal antibody drugs 61:110 manufactured recombinant monoclonal antibody drugs 61:110 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 0 3 from method 9:14 arg1 drugs 106:110 manufactured recombinant monoclonal antibody drugs 61:110 manufactured recombinant monoclonal antibody drugs 61:110 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 8 4 theme retention 1215:1223 arg1 times 1225:1229 LC retention times 1212:1229 LC retention times 1212:1229 The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses. 24828102 7 5 theme different 1051:1059 arg1 antibodies 1061:1070 different antibodies 1051:1070 different antibodies 1051:1070 The results showed that most of the N-glycans between different antibodies are nearly the same with different abundances. 24828102 3 6 theme rMab 552:555 arg1 drugs 557:561 eight commercial rMab drugs 535:561 eight commercial rMab drugs 535:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 1 7 theme many 257:260 arg1 attributes 262:271 the drugs' many attributes 246:271 the drugs' many attributes 246:271 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 2 8 theme intricate 327:335 arg1 structures 337:346 the intricate structures 323:346 the intricate structures 323:346 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 1 9 gly glycosylation 117:129 arg1 drugs 173:177 recombinant monoclonal antibody (rMab) drugs 134:177 recombinant monoclonal antibody (rMab) drugs 134:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 7 10 theme different 1097:1105 arg1 abundances 1107:1116 different abundances 1097:1116 different abundances 1097:1116 The results showed that most of the N-glycans between different antibodies are nearly the same with different abundances. 24828102 1 11 theme major 184:188 arg1 concern 190:196 a major concern 182:196 a major concern in the biopharmaceutical industry 182:230 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 1 11 theme major 184:188 arg1 glycosylation 117:129 The glycosylation 113:129 The glycosylation in recombinant monoclonal antibody (rMab) drugs 113:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 8 12 theme accurate 1235:1242 arg1 masses 1244:1249 accurate masses 1235:1249 accurate masses 1235:1249 The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses. 24828102 4 13 theme structures 585:594 arg1 library 566:572 A library 564:572 A library of over 70 structures 564:594 A library of over 70 structures was developed for the rapid characterization of rMab. 24828102 3 14 theme commercial 541:550 arg1 drugs 557:561 eight commercial rMab drugs 535:561 eight commercial rMab drugs 535:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 2 15 theme tools 400:404 arg1 microheterogeneity 349:366 microheterogeneity 349:366 microheterogeneity 349:366 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 2 15 theme tools 400:404 arg1 structures 337:346 the intricate structures 323:346 the intricate structures 323:346 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 2 15 theme tools 400:404 arg1 limitations 377:387 the limitations 373:387 the limitations of current tools for structural analysis 373:428 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 5 16 theme quadrupole 797:806 arg1 time-of-flight 808:821 an electrospray ionization hybrid quadrupole time-of-flight 763:821 an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time 763:856 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 16 theme quadrupole 797:806 arg1 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 8 17 theme library 1139:1145 arg1 utility 1123:1129 The utility 1119:1129 The utility of this library 1119:1145 The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses. 24828102 5 18 theme porous 680:685 arg1 column 712:717 a porous graphitized carbon (PGC) column 678:717 a porous graphitized carbon (PGC) column incorporated on a chip 678:740 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 2 19 theme current 392:398 arg1 tools 400:404 current tools 392:404 current tools for structural analysis 392:428 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 0 20 theme In-depth 0:7 arg1 method 9:14 In-depth method 0:14 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 0:111 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 6 21 theme exoglycosidase 970:983 arg1 sequencing 985:994 exoglycosidase sequencing 970:994 exoglycosidase sequencing 970:994 The complete structures were obtained through exoglycosidase sequencing. 24828102 3 22 theme liquid 461:466 arg1 LC-MS 502:506 LC-MS 502:506 LC-MS 502:506 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 3 22 theme liquid 461:466 arg1 spectrometry 488:499 liquid chromatography-mass spectrometry 461:499 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs 459:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 5 23 theme PGC 707:709 arg1 column 712:717 a porous graphitized carbon (PGC) column 678:717 a porous graphitized carbon (PGC) column incorporated on a chip 678:740 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 3 24 theme chromatography-mass 468:486 arg1 LC-MS 502:506 LC-MS 502:506 LC-MS 502:506 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 3 24 theme chromatography-mass 468:486 arg1 spectrometry 488:499 liquid chromatography-mass spectrometry 461:499 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs 459:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 8 25 theme LC 1212:1213 arg1 times 1225:1229 LC retention times 1212:1229 LC retention times 1212:1229 The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses. 24828102 1 26 theme biopharmaceutical 205:221 arg1 industry 223:230 the biopharmaceutical industry 201:230 the biopharmaceutical industry 201:230 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 5 27 theme electrospray 766:777 arg1 time-of-flight 808:821 an electrospray ionization hybrid quadrupole time-of-flight 763:821 an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time 763:856 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 27 theme electrospray 766:777 arg1 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 28 theme accurate 862:869 arg1 mass 871:874 accurate mass 862:874 accurate mass for each N-glycan 862:892 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 29 theme retention 843:851 arg1 time 853:856 The retention time 839:856 The retention time 839:856 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 2 30 theme structural 410:419 arg1 analysis 421:428 structural analysis 410:428 structural analysis 410:428 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 5 31 theme ionization 779:788 arg1 time-of-flight 808:821 an electrospray ionization hybrid quadrupole time-of-flight 763:821 an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time 763:856 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 31 theme ionization 779:788 arg1 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 6 32 theme complete 928:935 arg1 structures 937:946 The complete structures 924:946 The complete structures 924:946 The complete structures were obtained through exoglycosidase sequencing. 24828102 5 33 theme hybrid 790:795 arg1 time-of-flight 808:821 an electrospray ionization hybrid quadrupole time-of-flight 763:821 an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time 763:856 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 5 33 theme hybrid 790:795 arg1 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 ESI-Q-TOF 824:832 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 7 34 with same 1087:1090 arg1 abundances 1107:1116 different abundances 1097:1116 different abundances 1097:1116 The results showed that most of the N-glycans between different antibodies are nearly the same with different abundances. 24828102 0 35 theme glycosylation 44:56 arg1 characterization 24:39 the characterization 20:39 the characterization of glycosylation 20:56 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 2 36 gly microheterogeneity 349:366 arg1 tools 400:404 current tools 392:404 current tools for structural analysis 392:428 Characterization is important but complicated by the intricate structures, microheterogeneity, and the limitations of current tools for structural analysis. 24828102 1 37 theme recombinant 134:144 arg1 drugs 173:177 recombinant monoclonal antibody (rMab) drugs 134:177 recombinant monoclonal antibody (rMab) drugs 134:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 5 38 theme graphitized 687:697 arg1 column 712:717 a porous graphitized carbon (PGC) column 678:717 a porous graphitized carbon (PGC) column incorporated on a chip 678:740 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 1 39 theme monoclonal 146:155 arg1 rMab 167:170 rMab 167:170 rMab 167:170 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 1 39 theme monoclonal 146:155 arg1 antibody 157:164 monoclonal antibody 146:164 recombinant monoclonal antibody (rMab) drugs 134:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 1 40 from glycosylation 117:129 arg1 drugs 173:177 recombinant monoclonal antibody (rMab) drugs 134:177 recombinant monoclonal antibody (rMab) drugs 134:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 5 41 theme carbon 699:704 arg1 column 712:717 a porous graphitized carbon (PGC) column 678:717 a porous graphitized carbon (PGC) column incorporated on a chip 678:740 N-Glycans were separated on a porous graphitized carbon (PGC) column incorporated on a chip and then analyzed by an electrospray ionization hybrid quadrupole time-of-flight (ESI-Q-TOF) MS. The retention time and accurate mass for each N-glycan were recorded in the library. 24828102 1 42 theme antibody 157:164 arg1 drugs 173:177 recombinant monoclonal antibody (rMab) drugs 134:177 recombinant monoclonal antibody (rMab) drugs 134:177 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 3 43 theme spectrometry 488:499 arg1 library 518:524 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library 459:524 a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs 459:561 In this study, we developed a liquid chromatography-mass spectrometry (LC-MS) N-glycan library based on eight commercial rMab drugs. 24828102 0 44 theme recombinant 74:84 arg1 drugs 106:110 manufactured recombinant monoclonal antibody drugs 61:110 manufactured recombinant monoclonal antibody drugs 61:110 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 4 45 theme rMab 644:647 arg1 characterization 624:639 the rapid characterization 614:639 the rapid characterization of rMab 614:647 A library of over 70 structures was developed for the rapid characterization of rMab. 24828102 1 46 from concern 190:196 arg1 industry 223:230 the biopharmaceutical industry 201:230 the biopharmaceutical industry 201:230 The glycosylation in recombinant monoclonal antibody (rMab) drugs is a major concern in the biopharmaceutical industry as it impacts the drugs' many attributes. 24828102 4 47 theme rapid 618:622 arg1 characterization 624:639 the rapid characterization 614:639 the rapid characterization of rMab 614:647 A library of over 70 structures was developed for the rapid characterization of rMab. 24828102 0 48 theme manufactured 61:72 arg1 drugs 106:110 manufactured recombinant monoclonal antibody drugs 61:110 manufactured recombinant monoclonal antibody drugs 61:110 In-depth method for the characterization of glycosylation in manufactured recombinant monoclonal antibody drugs. 24828102 8 49 theme rapid 1187:1191 arg1 manner 1193:1198 a rapid manner 1185:1198 a rapid manner 1185:1198 The utility of this library enables one to identify structures in a rapid manner by matching LC retention times and accurate masses. 26082218 3 0 gly glycoprotein 443:454 arg1 glycoprotein 443:454 the glycoprotein 439:454 the glycoprotein 439:454 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 5 1 dep relationships.This 789:806 arg1 describes 816:824 describes 816:824 describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 816:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 5 2 theme glycoproteins 719:731 arg1 glycoforms 680:689 the appropriate glycoforms 664:689 the appropriate glycoforms of therapeutically relevant glycoproteins 664:731 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 6 3 theme glycosyltransferase 982:1000 arg1 disruption 954:963 the disruption 950:963 the disruption of an endogenous glycosyltransferase 950:1000 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 6 3 theme glycosyltransferase 982:1000 arg1 expression 1023:1032 the heterologous expression 1006:1032 the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host 1006:1135 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 7 4 theme transformation 1184:1197 arg1 procedure 1199:1207 the transformation procedure 1180:1207 the transformation procedure 1180:1207 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 7 4 theme transformation 1184:1197 arg1 screening 1222:1230 small-scale screening 1210:1230 small-scale screening 1210:1230 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 6 5 theme Endoplasmic 1090:1100 arg1 Reticulum 1102:1110 the Endoplasmic Reticulum 1086:1110 the Endoplasmic Reticulum 1086:1110 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 5 6 theme structure-function 770:787 arg1 relationships.This 789:806 structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 770:921 structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 770:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 4 7 theme N-glycosylation 531:545 arg1 pathway 547:553 the N-glycosylation pathway 527:553 the N-glycosylation pathway of Pichia pastoris 527:572 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 5 8 theme relationships.This 789:806 arg1 understanding 753:765 a better understanding 744:765 a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 744:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 5 9 theme relevant 710:717 arg1 glycoproteins 719:731 therapeutically relevant glycoproteins 694:731 therapeutically relevant glycoproteins 694:731 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 5 10 theme pivotal 633:639 arg1 importance 641:650 pivotal importance 633:650 pivotal importance 633:650 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 8 11 theme humanized 1574:1582 arg1 N-glycans 1584:1592 humanized N-glycans 1574:1592 humanized N-glycans 1574:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 5 12 theme glyco-engineered 857:872 arg1 strains 890:896 such glyco-engineered Pichia pastoris strains 852:896 such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 852:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 7 13 theme small-scale 1210:1220 arg1 procedure 1199:1207 the transformation procedure 1180:1207 the transformation procedure 1180:1207 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 7 13 theme small-scale 1210:1220 arg1 screening 1222:1230 small-scale screening 1210:1230 small-scale screening 1210:1230 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 1 14 theme Pichia 90:95 arg1 host 122:125 an important host 109:125 an important host for recombinant protein production 109:160 Pichia pastoris is an important host for recombinant protein production. 26082218 1 14 theme Pichia 90:95 arg1 pastoris 97:104 Pichia pastoris 90:104 Pichia pastoris 90:104 Pichia pastoris is an important host for recombinant protein production. 26082218 5 15 gly glycoforms 680:689 arg1 glycoproteins 719:731 therapeutically relevant glycoproteins 694:731 therapeutically relevant glycoproteins 694:731 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 6 16 theme glycosidase 1039:1049 arg1 disruption 954:963 the disruption 950:963 the disruption of an endogenous glycosyltransferase 950:1000 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 6 16 theme glycosidase 1039:1049 arg1 expression 1023:1032 the heterologous expression 1006:1032 the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host 1006:1135 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 8 17 theme iterative 1471:1479 arg1 fashion 1481:1487 an iterative fashion 1468:1487 an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans 1468:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 3 18 theme rapid 420:424 arg1 clearance 426:434 the rapid clearance 416:434 the rapid clearance of the glycoprotein from circulation 416:471 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 5 19 theme appropriate 668:678 arg1 glycoforms 680:689 the appropriate glycoforms 664:689 the appropriate glycoforms of therapeutically relevant glycoproteins 664:731 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 8 20 theme heterologous 1547:1558 arg1 proteins 1560:1567 heterologous proteins 1547:1567 heterologous proteins with humanized N-glycans 1547:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 0 21 theme pastoris 23:30 arg1 Pathway 48:54 the Pichia pastoris N-Glycosylation Pathway 12:54 the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology 12:87 Engineering the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology. 26082218 6 22 theme glycosyltransferase 1054:1072 arg1 disruption 954:963 the disruption 950:963 the disruption of an endogenous glycosyltransferase 950:1000 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 6 22 theme glycosyltransferase 1054:1072 arg1 expression 1023:1032 the heterologous expression 1006:1032 the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host 1006:1135 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 0 23 theme Pichia 16:21 arg1 pastoris 23:30 Pichia pastoris 16:30 the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology 12:87 Engineering the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology. 26082218 4 24 theme N-glycosylation 593:607 arg1 pathway 609:615 the human N-glycosylation pathway 583:615 the human N-glycosylation pathway 583:615 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 5 25 theme better 746:751 arg1 understanding 753:765 a better understanding 744:765 a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 744:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 1 26 theme important 112:120 arg1 host 122:125 an important host 109:125 an important host for recombinant protein production 109:160 Pichia pastoris is an important host for recombinant protein production. 26082218 1 26 theme important 112:120 arg1 pastoris 97:104 Pichia pastoris 90:104 Pichia pastoris 90:104 Pichia pastoris is an important host for recombinant protein production. 26082218 5 27 theme such 852:855 arg1 strains 890:896 such glyco-engineered Pichia pastoris strains 852:896 such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 852:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 4 28 theme human 587:591 arg1 pathway 609:615 the human N-glycosylation pathway 583:615 the human N-glycosylation pathway 583:615 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 4 29 theme recent 477:482 arg1 years 484:488 recent years 477:488 recent years 477:488 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 7 30 with clones 1360:1365 arg1 profile 1404:1410 the appropriate N-glycosylation profile 1372:1410 the appropriate N-glycosylation profile 1372:1410 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 6 31 theme host 1132:1135 arg1 Reticulum 1102:1110 the Endoplasmic Reticulum 1086:1110 the Endoplasmic Reticulum 1086:1110 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 6 31 theme host 1132:1135 arg1 Golgi 1119:1123 the Golgi 1115:1123 the Golgi of the host 1115:1135 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 0 32 theme N-Glycosylation 32:46 arg1 Pathway 48:54 the Pichia pastoris N-Glycosylation Pathway 12:54 the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology 12:87 Engineering the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology. 26082218 4 33 theme Pichia 558:563 arg1 pastoris 565:572 Pichia pastoris 558:572 Pichia pastoris 558:572 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 3 34 theme glycoprotein 443:454 arg1 clearance 426:434 the rapid clearance 416:434 the rapid clearance of the glycoprotein from circulation 416:471 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 2 35 theme production 176:185 arg1 platform 187:194 a protein production platform 166:194 a protein production platform 166:194 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 2 35 theme production 176:185 arg1 development 205:215 further development 197:215 further development for therapeutic glycoproteins 197:245 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 6 36 theme heterologous 1010:1021 arg1 expression 1023:1032 the heterologous expression 1006:1032 the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host 1006:1135 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 2 37 gly glycoproteins 233:245 arg1 glycoproteins 233:245 therapeutic glycoproteins 221:245 therapeutic glycoproteins 221:245 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 7 38 theme Fluorophore-Assisted 1288:1307 arg1 DSA-FACE 1336:1343 DSA-FACE 1336:1343 DSA-FACE 1336:1343 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 7 38 theme Fluorophore-Assisted 1288:1307 arg1 Electrophoresis 1319:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis 1268:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) 1268:1344 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 3 39 theme Such 333:336 arg1 N-glycans 338:346 Such N-glycans 333:346 Such N-glycans 333:346 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 2 40 theme protein 168:174 arg1 platform 187:194 a protein production platform 166:194 a protein production platform 166:194 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 2 40 theme protein 168:174 arg1 development 205:215 further development 197:215 further development for therapeutic glycoproteins 197:245 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 7 41 theme appropriate 1376:1386 arg1 profile 1404:1410 the appropriate N-glycosylation profile 1372:1410 the appropriate N-glycosylation profile 1372:1410 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 5 42 theme pastoris 881:888 arg1 strains 890:896 such glyco-engineered Pichia pastoris strains 852:896 such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®) 852:921 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 7 43 theme DNA-Sequencer-Aided 1268:1286 arg1 DSA-FACE 1336:1343 DSA-FACE 1336:1343 DSA-FACE 1336:1343 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 7 43 theme DNA-Sequencer-Aided 1268:1286 arg1 Electrophoresis 1319:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis 1268:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) 1268:1344 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 2 44 theme therapeutic 221:231 arg1 glycoproteins 233:245 therapeutic glycoproteins 221:245 therapeutic glycoproteins 221:245 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 8 45 theme pastoris 1527:1534 arg1 clones 1510:1515 clones 1510:1515 clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans 1510:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 4 46 theme much 491:494 arg1 effort 496:501 much effort 491:501 much effort 491:501 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 4 47 theme pastoris 565:572 arg1 pathway 547:553 the N-glycosylation pathway 527:553 the N-glycosylation pathway of Pichia pastoris 527:572 In recent years, much effort has gone to engineering the N-glycosylation pathway of Pichia pastoris to mimic the human N-glycosylation pathway. 26082218 6 48 dep glycosidase 1039:1049 arg1 a 1037:1037 a 1037:1037 a 1037:1037 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 8 49 theme Pichia 1520:1525 arg1 pastoris 1527:1534 Pichia pastoris 1520:1534 Pichia pastoris expressing heterologous proteins with humanized N-glycans 1520:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 1 50 theme recombinant 131:141 arg1 production 151:160 recombinant protein production 131:160 recombinant protein production 131:160 Pichia pastoris is an important host for recombinant protein production. 26082218 2 51 theme common 306:311 arg1 N-glycosylation 290:304 the high-mannose-type N-glycosylation 268:304 the high-mannose-type N-glycosylation common to yeast and fungi 268:330 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 2 52 theme high-mannose-type 272:288 arg1 N-glycosylation 290:304 the high-mannose-type N-glycosylation 268:304 the high-mannose-type N-glycosylation common to yeast and fungi 268:330 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 1 53 theme protein 143:149 arg1 production 151:160 recombinant protein production 131:160 recombinant protein production 131:160 Pichia pastoris is an important host for recombinant protein production. 26082218 8 54 with proteins 1560:1567 arg1 N-glycans 1584:1592 humanized N-glycans 1574:1592 humanized N-glycans 1574:1592 The steps described in this chapter can be followed in an iterative fashion in order to generate clones of Pichia pastoris expressing heterologous proteins with humanized N-glycans. 26082218 3 55 theme downstream 363:372 arg1 processing 374:383 downstream processing 363:383 downstream processing 363:383 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 2 56 theme further 197:203 arg1 platform 187:194 a protein production platform 166:194 a protein production platform 166:194 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 2 56 theme further 197:203 arg1 development 205:215 further development 197:215 further development for therapeutic glycoproteins 197:245 As a protein production platform, further development for therapeutic glycoproteins has been hindered by the high-mannose-type N-glycosylation common to yeast and fungi. 26082218 7 57 from step 1147:1150 arg1 process 1159:1165 the process 1155:1165 the process 1155:1165 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 3 58 from circulation 461:471 arg1 clearance 426:434 the rapid clearance 416:434 the rapid clearance of the glycoprotein from circulation 416:471 Such N-glycans can complicate downstream processing, might be immunogenic or cause the rapid clearance of the glycoprotein from circulation. 26082218 6 59 theme endogenous 971:980 arg1 glycosyltransferase 982:1000 an endogenous glycosyltransferase 968:1000 an endogenous glycosyltransferase 968:1000 This strategy consists of the disruption of an endogenous glycosyltransferase and the heterologous expression of a glycosidase or glycosyltransferase targeted to the Endoplasmic Reticulum or the Golgi of the host. 26082218 0 60 theme GlycoSwitch 66:76 arg1 Technology 78:87 the GlycoSwitch Technology 62:87 the GlycoSwitch Technology 62:87 Engineering the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology. 26082218 7 61 theme N-glycosylation 1388:1402 arg1 profile 1404:1410 the appropriate N-glycosylation profile 1372:1410 the appropriate N-glycosylation profile 1372:1410 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 5 62 gly glycoproteins 719:731 arg1 glycoproteins 719:731 therapeutically relevant glycoproteins 694:731 therapeutically relevant glycoproteins 694:731 This can be of pivotal importance to generate the appropriate glycoforms of therapeutically relevant glycoproteins or to gain a better understanding of structure-function relationships.This chapter describes the methodology to create such glyco-engineered Pichia pastoris strains using the GlycoSwitch(®). 26082218 7 63 theme Capillary 1309:1317 arg1 DSA-FACE 1336:1343 DSA-FACE 1336:1343 DSA-FACE 1336:1343 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 26082218 7 63 theme Capillary 1309:1317 arg1 Electrophoresis 1319:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis 1268:1333 DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) 1268:1344 For each step in the process, we describe the transformation procedure, small-scale screening and we also describe how to perform DNA-Sequencer-Aided Fluorophore-Assisted Capillary Electrophoresis (DSA-FACE) to select for clones with the appropriate N-glycosylation profile. 25124522 5 0 theme core 851:854 arg1 oxazoline 867:875 the N-glycan core Man3GlcNAc oxazoline 838:875 the N-glycan core Man3GlcNAc oxazoline 838:875 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 6 1 theme chemoenzymatic 1162:1175 arg1 synthesis 1177:1185 endo-glycosidase-enabled chemoenzymatic synthesis 1137:1185 endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1137:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 6 2 theme glycoconjugates 1210:1224 arg1 synthesis 1177:1185 endo-glycosidase-enabled chemoenzymatic synthesis 1137:1185 endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1137:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 4 3 theme sophisticated 759:771 arg1 chromatography 773:786 sophisticated chromatography 759:786 sophisticated chromatography 759:786 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 5 4 theme Man3GlcNAc 856:865 arg1 oxazoline 867:875 the N-glycan core Man3GlcNAc oxazoline 838:875 the N-glycan core Man3GlcNAc oxazoline 838:875 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 1 5 theme glycan 164:169 arg1 structures 171:180 glycan structures 164:180 glycan structures 164:180 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 6 6 theme homogeneous 1198:1208 arg1 glycoconjugates 1210:1224 various homogeneous glycoconjugates 1190:1224 various homogeneous glycoconjugates 1190:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 4 7 theme egg 661:663 arg1 powder 670:675 the egg yolk powder 657:675 the egg yolk powder 657:675 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 0 8 theme egg 82:84 arg1 yolks 86:90 egg yolks 82:90 egg yolks 82:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 2 9 theme natural 332:338 arg1 resource 340:347 natural resource 332:347 natural resource of homogeneous glycans 332:370 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 4 10 theme reported 791:798 arg1 methods 800:806 reported methods 791:806 reported methods 791:806 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 0 11 theme subsequent 96:105 arg1 semi-synthesis 107:120 subsequent semi-synthesis 96:120 subsequent semi-synthesis of Man3GlcNAc oxazoline 96:144 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 1 12 from Heterogeneity 147:159 arg1 glycoconjugates 192:206 native glycoconjugates 185:206 native glycoconjugates 185:206 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 3 13 theme gram-scale 471:480 arg1 production 482:491 gram-scale production 471:491 gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks 471:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 1 14 theme structures 171:180 arg1 Heterogeneity 147:159 Heterogeneity 147:159 Heterogeneity of glycan structures in native glycoconjugates 147:206 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 4 15 theme yolk 665:668 arg1 powder 670:675 the egg yolk powder 657:675 the egg yolk powder 657:675 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 3 16 theme sialylglycopeptide 496:513 arg1 production 482:491 gram-scale production 471:491 gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks 471:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 0 17 theme simplified 2:11 arg1 procedure 13:21 A simplified procedure 0:21 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks 0:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 4 18 theme extraction 626:635 arg1 process 637:643 the extraction process 622:643 the extraction process 622:643 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 1 19 theme native 185:190 arg1 glycoconjugates 192:206 native glycoconjugates 185:206 native glycoconjugates 185:206 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 6 20 theme chemical 1043:1050 arg1 approach 1052:1059 the pure chemical approach 1034:1059 the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1034:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 5 21 theme oxazoline 956:964 arg1 formation 966:974 successive oxazoline formation 945:974 successive oxazoline formation 945:974 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 2 22 theme glycans 364:370 arg1 resource 340:347 natural resource 332:347 natural resource of homogeneous glycans 332:370 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 2 23 theme homogeneous 352:362 arg1 glycans 364:370 homogeneous glycans 352:370 homogeneous glycans 352:370 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 6 24 theme total 1086:1090 arg1 synthesis 1092:1100 multi-step total synthesis 1075:1100 multi-step total synthesis 1075:1100 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 6 25 theme endo-glycosidase-enabled 1137:1160 arg1 synthesis 1177:1185 endo-glycosidase-enabled chemoenzymatic synthesis 1137:1185 endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1137:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 4 26 theme % 684:684 arg1 acetone 686:692 40% acetone 682:692 40% acetone 682:692 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 5 27 theme oxazoline 867:875 arg1 semi-synthesis 820:833 Subsequent semi-synthesis 809:833 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP 809:884 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 3 28 theme complex-type 555:566 arg1 N-glycan 568:575 a disialyl biantennary complex-type N-glycan 532:575 a disialyl biantennary complex-type N-glycan 532:575 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 0 29 theme oxazoline 136:144 arg1 procedure 13:21 A simplified procedure 0:21 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks 0:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 0 29 theme oxazoline 136:144 arg1 semi-synthesis 107:120 subsequent semi-synthesis 96:120 subsequent semi-synthesis of Man3GlcNAc oxazoline 96:144 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 6 30 theme efficient 982:990 arg1 semi-synthesis 992:1005 This efficient semi-synthesis 977:1005 This efficient semi-synthesis 977:1005 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 6 31 theme multi-step 1075:1084 arg1 synthesis 1092:1100 multi-step total synthesis 1075:1100 multi-step total synthesis 1075:1100 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 1 32 theme precise 223:229 arg1 studies 231:237 precise studies 223:237 precise studies on carbohydrate-involved biological functions 223:283 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 0 33 theme gram-scale 27:36 arg1 production 38:47 gram-scale production 27:47 gram-scale production of sialylglycopeptide (SGP) from egg yolks 27:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 0 34 theme Man3GlcNAc 125:134 arg1 oxazoline 136:144 Man3GlcNAc oxazoline 125:144 Man3GlcNAc oxazoline 125:144 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 3 35 contain containing 521:530 arg1 SGP 516:518 SGP 516:518 SGP 516:518 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 3 35 contain containing 521:530 arg1 sialylglycopeptide 496:513 sialylglycopeptide 496:513 sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks 496:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 3 35 contain containing 521:530 arg2 N-glycan 568:575 a disialyl biantennary complex-type N-glycan 532:575 a disialyl biantennary complex-type N-glycan 532:575 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 4 36 theme 40 682:683 arg1 % 684:684 % 684:684 % 684:684 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 5 37 theme successive 945:954 arg1 formation 966:974 successive oxazoline formation 945:974 successive oxazoline formation 945:974 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 0 38 theme sialylglycopeptide 52:69 arg1 production 38:47 gram-scale production 27:47 gram-scale production of sialylglycopeptide (SGP) from egg yolks 27:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 4 39 theme high-speed 728:737 arg1 centrifugation 739:752 high-speed centrifugation 728:752 high-speed centrifugation 728:752 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 1 40 gly Heterogeneity 147:159 arg1 structures 171:180 glycan structures 164:180 glycan structures 164:180 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 1 41 from studies 231:237 arg1 functions 275:283 carbohydrate-involved biological functions 242:283 carbohydrate-involved biological functions 242:283 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 3 42 theme biantennary 543:553 arg1 N-glycan 568:575 a disialyl biantennary complex-type N-glycan 532:575 a disialyl biantennary complex-type N-glycan 532:575 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 3 43 theme optimized 447:455 arg1 procedure 457:465 an optimized procedure 444:465 an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks 444:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 6 44 theme pure 1038:1041 arg1 approach 1052:1059 the pure chemical approach 1034:1059 the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1034:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 2 45 theme practical 387:395 arg1 approach 397:404 a practical approach 385:404 a practical approach to solve this problem 385:426 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 4 46 theme new 597:599 arg1 procedure 601:609 Our new procedure 593:609 Our new procedure 593:609 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 3 47 gly sialylglycopeptide 496:513 arg2 SGP 516:518 SGP 516:518 SGP 516:518 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 3 47 gly sialylglycopeptide 496:513 arg2 sialylglycopeptide 496:513 sialylglycopeptide 496:513 sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks 496:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 2 48 theme homogeneous 299:309 arg1 glycoconjugates 311:325 homogeneous glycoconjugates 299:325 homogeneous glycoconjugates from natural resource of homogeneous glycans 299:370 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 6 49 theme synthesis 1177:1185 arg1 application 1122:1132 the application 1118:1132 the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates 1118:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 0 50 from yolks 86:90 arg1 production 38:47 gram-scale production 27:47 gram-scale production of sialylglycopeptide (SGP) from egg yolks 27:90 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 3 51 theme disialyl 534:541 arg1 N-glycan 568:575 a disialyl biantennary complex-type N-glycan 532:575 a disialyl biantennary complex-type N-glycan 532:575 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 2 52 from resource 340:347 arg1 glycoconjugates 311:325 homogeneous glycoconjugates 299:325 homogeneous glycoconjugates from natural resource of homogeneous glycans 299:370 To construct homogeneous glycoconjugates from natural resource of homogeneous glycans is therefore a practical approach to solve this problem. 25124522 1 53 theme carbohydrate-involved 242:262 arg1 functions 275:283 carbohydrate-involved biological functions 242:283 carbohydrate-involved biological functions 242:283 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 6 54 theme various 1190:1196 arg1 glycoconjugates 1210:1224 various homogeneous glycoconjugates 1190:1224 various homogeneous glycoconjugates 1190:1224 This efficient semi-synthesis provides an alternative to the pure chemical approach that involves multi-step total synthesis and facilitates the application of endo-glycosidase-enabled chemoenzymatic synthesis of various homogeneous glycoconjugates. 25124522 3 55 theme egg 582:584 arg1 yolks 586:590 egg yolks 582:590 egg yolks 582:590 We report here an optimized procedure for gram-scale production of sialylglycopeptide (SGP) containing a disialyl biantennary complex-type N-glycan from egg yolks. 25124522 4 56 theme massive 704:710 arg1 emulsification 712:725 massive emulsification 704:725 massive emulsification 704:725 Our new procedure simplified the extraction process by treating the egg yolk powder with 40% acetone, avoiding massive emulsification, high-speed centrifugation, and sophisticated chromatography in reported methods. 25124522 1 57 theme biological 264:273 arg1 functions 275:283 carbohydrate-involved biological functions 242:283 carbohydrate-involved biological functions 242:283 Heterogeneity of glycan structures in native glycoconjugates always hampers precise studies on carbohydrate-involved biological functions. 25124522 5 58 theme N-glycan 842:849 arg1 oxazoline 867:875 the N-glycan core Man3GlcNAc oxazoline 838:875 the N-glycan core Man3GlcNAc oxazoline 838:875 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 5 59 from SGP 882:884 arg1 semi-synthesis 820:833 Subsequent semi-synthesis 809:833 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP 809:884 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25124522 0 60 gly sialylglycopeptide 52:69 arg2 sialylglycopeptide 52:69 sialylglycopeptide 52:69 sialylglycopeptide (SGP) 52:75 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 0 60 gly sialylglycopeptide 52:69 arg2 SGP 72:74 SGP 72:74 SGP 72:74 A simplified procedure for gram-scale production of sialylglycopeptide (SGP) from egg yolks and subsequent semi-synthesis of Man3GlcNAc oxazoline. 25124522 5 61 theme Subsequent 809:818 arg1 semi-synthesis 820:833 Subsequent semi-synthesis 809:833 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP 809:884 Subsequent semi-synthesis of the N-glycan core Man3GlcNAc oxazoline from SGP was accomplished for the first-time via glyco-trimming and successive oxazoline formation. 25698222 11 0 theme glycans 1605:1611 arg1 quantitation 1589:1600 quantitation 1589:1600 quantitation 1589:1600 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 0 theme glycans 1605:1611 arg1 identification 1570:1583 identification 1570:1583 identification 1570:1583 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 10 1 theme N-glycans 1358:1366 arg1 expression 1344:1353 the expression 1340:1353 the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231) 1340:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 11 2 attach derived 1613:1619 arg2 glycans 1605:1611 glycans 1605:1611 glycans derived from glycoproteins purified or present in complex biological samples 1605:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 2 attach derived 1613:1619 arg1 glycoproteins 1626:1638 glycoproteins 1626:1638 glycoproteins purified or present in complex biological samples 1626:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 1 3 theme methods 275:281 arg1 development 227:237 the development 223:237 the development of sensitive quantitative glycomics methods 223:281 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 11 4 theme present 1652:1658 arg1 glycoproteins 1626:1638 glycoproteins 1626:1638 glycoproteins purified or present in complex biological samples 1626:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 9 5 theme level 1149:1153 arg1 100th 1135:1139 the 100th 1131:1139 the 100th of a μL level 1131:1153 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 11 6 theme purified 1640:1647 arg1 glycoproteins 1626:1638 glycoproteins 1626:1638 glycoproteins purified or present in complex biological samples 1626:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 7 theme N-glycan 1532:1539 arg1 method 1508:1513 the described MRM method 1490:1513 the described MRM method of permethylated N-glycan 1490:1539 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 6 8 theme reliable 851:858 arg1 quantitation 860:871 reliable quantitation 851:871 reliable quantitation 851:871 Additionally, the use of three transitions was shown to facilitate reliable quantitation. 25698222 8 9 theme Reliable 972:979 arg1 detection 981:989 detection 981:989 detection 981:989 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 11 10 theme MRM 1504:1506 arg1 method 1508:1513 the described MRM method 1490:1513 the described MRM method of permethylated N-glycan 1490:1539 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 11 gly glycoproteins 1626:1638 arg1 glycoproteins 1626:1638 glycoproteins 1626:1638 glycoproteins purified or present in complex biological samples 1626:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 9 12 theme pooled 1185:1190 arg1 serum 1204:1208 pooled human blood serum 1185:1208 pooled human blood serum 1185:1208 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 2 13 theme low 320:322 arg1 abundance 324:332 low abundance 320:332 low abundance 320:332 Quantitation of glycans existing at low abundance is still analytically challenging. 25698222 9 14 theme blood 1198:1202 arg1 serum 1204:1208 pooled human blood serum 1185:1208 pooled human blood serum 1185:1208 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 11 15 from present 1652:1658 arg1 samples 1682:1688 complex biological samples 1663:1688 complex biological samples 1663:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 7 16 from compositions 897:908 arg1 serum 925:929 human blood serum 913:929 human blood serum 913:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 1 17 theme glycans 127:133 arg1 implications 145:156 their implications 139:156 their implications in disease development and progression 139:195 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 1 17 theme glycans 127:133 arg1 roles 118:122 The important biological roles 93:122 The important biological roles of glycans 93:133 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 7 18 from serum 925:929 arg1 total 876:880 A total 874:880 A total of 88 N-glycan compositions in human blood serum 874:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 10 19 attach derived 1368:1374 arg1 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 19 attach derived 1368:1374 arg1 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 19 attach derived 1368:1374 arg2 N-glycans 1358:1366 N-glycans 1358:1366 N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231) 1358:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 19 attach derived 1368:1374 arg1 cells 1414:1418 brain-targeting breast carcinoma cells 1381:1418 brain-targeting breast carcinoma cells (MDA-MB-231BR) 1381:1433 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 19 attach derived 1368:1374 arg1 cells 1464:1468 metastatic breast cancer cells 1439:1468 metastatic breast cancer cells (MDA-MB-231) 1439:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 9 20 theme concentration 1230:1242 arg1 range 1244:1248 a dynamic concentration range 1220:1248 a dynamic concentration range of three orders of magnitude 1220:1277 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 4 21 theme collision 533:541 arg1 energy 543:548 Optimum normalized collision energy 514:548 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan 514:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 21 theme collision 533:541 arg1 CE 551:552 CE 551:552 CE 551:552 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 9 22 theme orders 1259:1264 arg1 range 1244:1248 a dynamic concentration range 1220:1248 a dynamic concentration range of three orders of magnitude 1220:1277 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 8 23 theme glycans 1017:1023 arg1 detection 981:989 detection 981:989 detection 981:989 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 8 23 theme glycans 1017:1023 arg1 quantitation 995:1006 quantitation 995:1006 quantitation 995:1006 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 1 24 from implications 145:156 arg1 progression 185:195 progression 185:195 progression 185:195 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 1 24 from implications 145:156 arg1 development 169:179 disease development 161:179 disease development 161:179 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 11 25 from samples 1682:1688 arg1 present 1652:1658 present 1652:1658 present 1652:1658 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 10 26 theme breast 1450:1455 arg1 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 26 theme breast 1450:1455 arg1 cells 1464:1468 metastatic breast cancer cells 1439:1468 metastatic breast cancer cells (MDA-MB-231) 1439:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 5 27 theme complex 736:742 arg1 N-glycan 749:756 complex type N-glycan 736:756 complex type N-glycan 736:756 The optimum CE for mannose and complex type N-glycan was determined to be 35%. 25698222 9 28 theme magnitude 1269:1277 arg1 orders 1259:1264 three orders 1253:1264 three orders of magnitude 1253:1277 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 7 29 theme human 913:917 arg1 serum 925:929 human blood serum 913:929 human blood serum 913:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 3 30 theme quantitation 404:415 arg1 method 417:422 an N-linked glycans quantitation method 384:422 an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument 384:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 4 31 theme Optimum 514:520 arg1 energy 543:548 Optimum normalized collision energy 514:548 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan 514:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 31 theme Optimum 514:520 arg1 CE 551:552 CE 551:552 CE 551:552 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 7 32 from total 876:880 arg1 serum 925:929 human blood serum 913:929 human blood serum 913:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 7 33 theme compositions 897:908 arg1 total 876:880 A total 874:880 A total of 88 N-glycan compositions in human blood serum 874:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 1 34 theme important 97:105 arg1 roles 118:122 The important biological roles 93:122 The important biological roles of glycans 93:133 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 3 35 theme N-linked 387:394 arg1 glycans 396:402 N-linked glycans 387:402 an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument 384:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 0 36 theme permethylated 16:28 arg1 N-glycans 30:38 permethylated N-glycans 16:38 permethylated N-glycans 16:38 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 25698222 3 37 theme triple 470:475 arg1 instrument 488:497 a triple quadrupole instrument 468:497 a triple quadrupole instrument 468:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 11 38 theme biological 1671:1680 arg1 samples 1682:1688 complex biological samples 1663:1688 complex biological samples 1663:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 39 theme reliable 1561:1568 arg1 identification 1570:1583 identification 1570:1583 identification 1570:1583 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 0 40 theme monitoring 66:75 arg1 LC-MS/MS 83:90 multiple-reaction monitoring (MRM) LC-MS/MS 48:90 multiple-reaction monitoring (MRM) LC-MS/MS 48:90 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 25698222 8 41 theme serum 1080:1084 arg1 0.005 μL 1062:1069 0.005 μL 1062:1069 0.005 μL of blood serum 1062:1084 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 6 42 theme transitions 815:825 arg1 use 802:804 the use 798:804 the use of three transitions 798:825 Additionally, the use of three transitions was shown to facilitate reliable quantitation. 25698222 10 43 used utilized 1305:1312 arg2 MRM 1280:1282 MRM 1280:1282 MRM 1280:1282 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 44 theme breast 1397:1402 arg1 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 44 theme breast 1397:1402 arg1 cells 1414:1418 brain-targeting breast carcinoma cells 1381:1418 brain-targeting breast carcinoma cells (MDA-MB-231BR) 1381:1433 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 11 45 theme rapid 1551:1555 arg1 identification 1570:1583 identification 1570:1583 identification 1570:1583 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 1 46 from roles 118:122 arg1 progression 185:195 progression 185:195 progression 185:195 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 1 46 from roles 118:122 arg1 development 169:179 disease development 161:179 disease development 161:179 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 1 47 theme sensitive 242:250 arg1 methods 275:281 sensitive quantitative glycomics methods 242:281 sensitive quantitative glycomics methods 242:281 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 1 48 theme glycomics 265:273 arg1 methods 275:281 sensitive quantitative glycomics methods 242:281 sensitive quantitative glycomics methods 242:281 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 9 49 theme μL 1146:1147 arg1 level 1149:1153 a μL level 1144:1153 a μL level 1144:1153 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 7 50 theme MRM 958:960 arg1 approach 962:969 this MRM approach 953:969 this MRM approach 953:969 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 11 51 theme described 1494:1502 arg1 method 1508:1513 the described MRM method 1490:1513 the described MRM method of permethylated N-glycan 1490:1539 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 2 52 theme glycans 300:306 arg1 Quantitation 284:295 Quantitation 284:295 Quantitation of glycans existing at low abundance 284:332 Quantitation of glycans existing at low abundance is still analytically challenging. 25698222 10 53 theme brain-targeting 1381:1395 arg1 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 53 theme brain-targeting 1381:1395 arg1 cells 1414:1418 brain-targeting breast carcinoma cells 1381:1418 brain-targeting breast carcinoma cells (MDA-MB-231BR) 1381:1433 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 11 54 dep identification 1570:1583 arg1 a 1549:1549 a 1549:1549 a 1549:1549 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 55 theme permethylated 1518:1530 arg1 N-glycan 1532:1539 permethylated N-glycan 1518:1539 permethylated N-glycan 1518:1539 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 5 56 theme optimum 709:715 arg1 % 781:781 35% 779:781 35% 779:781 The optimum CE for mannose and complex type N-glycan was determined to be 35%. 25698222 5 56 theme optimum 709:715 arg1 CE 717:718 The optimum CE 705:718 The optimum CE for mannose and complex type N-glycan 705:756 The optimum CE for mannose and complex type N-glycan was determined to be 35%. 25698222 9 57 theme human 1192:1196 arg1 serum 1204:1208 pooled human blood serum 1185:1208 pooled human blood serum 1185:1208 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 4 58 gly fucosylated 668:678 arg1 N-glycans 694:702 either fucosylated or sialylated N-glycans 661:702 either fucosylated or sialylated N-glycans 661:702 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 59 gly fucosylated 579:589 arg1 N-glycan 591:598 both sialylated and fucosylated N-glycan 559:598 both sialylated and fucosylated N-glycan 559:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 60 gly sialylated 683:692 arg1 N-glycans 694:702 either fucosylated or sialylated N-glycans 661:702 either fucosylated or sialylated N-glycans 661:702 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 3 61 link N-linked 387:394 arg1 glycans 396:402 N-linked glycans 387:402 an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument 384:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 9 62 theme down 1123:1126 arg1 N-glycans 1113:1121 N-glycans 1113:1121 N-glycans down to the 100th of a μL level 1113:1153 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 5 63 theme type 744:747 arg1 N-glycan 749:756 complex type N-glycan 736:756 complex type N-glycan 736:756 The optimum CE for mannose and complex type N-glycan was determined to be 35%. 25698222 9 64 theme dynamic 1222:1228 arg1 range 1244:1248 a dynamic concentration range 1220:1248 a dynamic concentration range of three orders of magnitude 1220:1277 Accordingly, N-glycans down to the 100th of a μL level can be reliably quantified in pooled human blood serum, spanning a dynamic concentration range of three orders of magnitude. 25698222 11 65 attach present 1652:1658 arg1 samples 1682:1688 complex biological samples 1663:1688 complex biological samples 1663:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 11 65 attach present 1652:1658 arg2 glycoproteins 1626:1638 glycoproteins 1626:1638 glycoproteins purified or present in complex biological samples 1626:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 4 66 theme fucosylated 668:678 arg1 N-glycans 694:702 either fucosylated or sialylated N-glycans 661:702 either fucosylated or sialylated N-glycans 661:702 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 67 gly sialylated 564:573 arg1 N-glycan 591:598 both sialylated and fucosylated N-glycan 559:598 both sialylated and fucosylated N-glycan 559:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 1 68 theme disease 161:167 arg1 development 169:179 disease development 161:179 disease development 161:179 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 4 69 theme sialylated 683:692 arg1 N-glycans 694:702 either fucosylated or sialylated N-glycans 661:702 either fucosylated or sialylated N-glycans 661:702 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 10 70 theme carcinoma 1404:1412 arg1 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MDA-MB-231BR 1421:1432 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 70 theme carcinoma 1404:1412 arg1 cells 1414:1418 brain-targeting breast carcinoma cells 1381:1418 brain-targeting breast carcinoma cells (MDA-MB-231BR) 1381:1433 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 7 71 theme N-glycan 888:895 arg1 compositions 897:908 88 N-glycan compositions 885:908 88 N-glycan compositions in human blood serum 885:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 4 72 theme normalized 522:531 arg1 energy 543:548 Optimum normalized collision energy 514:548 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan 514:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 4 72 theme normalized 522:531 arg1 CE 551:552 CE 551:552 CE 551:552 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 10 73 theme metastatic 1439:1448 arg1 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 73 theme metastatic 1439:1448 arg1 cells 1464:1468 metastatic breast cancer cells 1439:1468 metastatic breast cancer cells (MDA-MB-231) 1439:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 7 74 theme blood 919:923 arg1 serum 925:929 human blood serum 913:929 human blood serum 913:929 A total of 88 N-glycan compositions in human blood serum were quantified using this MRM approach. 25698222 4 75 theme fucosylated 579:589 arg1 N-glycan 591:598 both sialylated and fucosylated N-glycan 559:598 both sialylated and fucosylated N-glycan 559:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 0 76 theme N-glycans 30:38 arg1 Quantitation 0:11 Quantitation 0:11 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 0:91 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 25698222 8 77 theme 0.005 μL 1062:1069 arg1 equivalence 1047:1057 the equivalence 1043:1057 the equivalence of 0.005 μL of blood serum 1043:1084 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 1 78 theme biological 107:116 arg1 roles 118:122 The important biological roles 93:122 The important biological roles of glycans 93:133 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 25698222 3 79 theme glycans 396:402 arg1 method 417:422 an N-linked glycans quantitation method 384:422 an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument 384:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 0 80 theme multiple-reaction 48:64 arg1 MRM 78:80 MRM 78:80 MRM 78:80 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 25698222 0 80 theme multiple-reaction 48:64 arg1 monitoring 66:75 multiple-reaction monitoring 48:75 multiple-reaction monitoring (MRM) LC-MS/MS 48:90 Quantitation of permethylated N-glycans through multiple-reaction monitoring (MRM) LC-MS/MS. 25698222 8 81 theme blood 1074:1078 arg1 serum 1080:1084 blood serum 1074:1084 blood serum 1074:1084 Reliable detection and quantitation of these glycans was achieved when the equivalence of 0.005 μL of blood serum was analyzed. 25698222 10 82 theme cancer 1457:1462 arg1 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MDA-MB-231 1471:1480 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 10 82 theme cancer 1457:1462 arg1 cells 1464:1468 metastatic breast cancer cells 1439:1468 metastatic breast cancer cells (MDA-MB-231) 1439:1481 MRM was also effectively utilized to quantitatively compare the expression of N-glycans derived from brain-targeting breast carcinoma cells (MDA-MB-231BR) and metastatic breast cancer cells (MDA-MB-231). 25698222 11 83 theme complex 1663:1669 arg1 samples 1682:1688 complex biological samples 1663:1688 complex biological samples 1663:1688 Thus, the described MRM method of permethylated N-glycan enables a rapid and reliable identification and quantitation of glycans derived from glycoproteins purified or present in complex biological samples. 25698222 3 84 theme quadrupole 477:486 arg1 instrument 488:497 a triple quadrupole instrument 468:497 a triple quadrupole instrument 468:497 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 3 85 theme multiple-reaction 430:446 arg1 MRM 460:462 MRM 460:462 MRM 460:462 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 3 85 theme multiple-reaction 430:446 arg1 monitoring 448:457 multiple-reaction monitoring 430:457 multiple-reaction monitoring (MRM) 430:463 In this study, an N-linked glycans quantitation method using multiple-reaction monitoring (MRM) on a triple quadrupole instrument was developed. 25698222 4 86 theme sialylated 564:573 arg1 N-glycan 591:598 both sialylated and fucosylated N-glycan 559:598 both sialylated and fucosylated N-glycan 559:598 Optimum normalized collision energy (CE) for both sialylated and fucosylated N-glycan was determined to be 30%, whereas it was found to be 35% for either fucosylated or sialylated N-glycans. 25698222 1 87 theme quantitative 252:263 arg1 methods 275:281 sensitive quantitative glycomics methods 242:281 sensitive quantitative glycomics methods 242:281 The important biological roles of glycans and their implications in disease development and progression have created a demand for the development of sensitive quantitative glycomics methods. 26800543 7 0 from spectra 994:1000 arg1 structures 968:977 glycan structures 961:977 glycan structures from HCD MS/MS spectra 961:1000 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 0 1 theme HCD 67:69 arg1 Spectra 71:77 HCD Spectra 67:77 HCD Spectra 67:77 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 4 2 theme structures 524:533 arg1 identification 499:512 the identification 495:512 the identification of glycan structures from mass spectral data 495:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 5 3 theme heuristic 659:667 arg1 algorithm 669:677 a heuristic algorithm 657:677 a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides 657:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 1 4 theme proteomics 148:157 arg1 study 159:163 proteomics study 148:163 proteomics study 148:163 Mass spectrometry has become a widely used analytical technique for proteomics study because of its high throughput and sensitivity. 26800543 5 5 from spectra 724:730 arg1 sequencing 698:707 glycan de novo sequencing 683:707 glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides 683:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 3 6 theme proteins 365:372 arg1 Glycosylation 291:303 Glycosylation 291:303 Glycosylation 291:303 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 6 theme proteins 365:372 arg1 relevant 383:390 relevant 383:390 relevant 383:390 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 6 theme proteins 365:372 arg1 modification 349:360 a frequently occurred post-translational modification 308:360 a frequently occurred post-translational modification of proteins which is relevant to humans' health 308:408 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 4 7 theme glycan 517:522 arg1 structures 524:533 glycan structures 517:533 glycan structures from mass spectral data 517:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 5 8 theme glycan 610:615 arg1 problem 636:642 the glycan de novo sequencing problem 606:642 the glycan de novo sequencing problem 606:642 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 2 9 theme glycan 273:278 arg1 structure 280:288 glycan structure 273:288 glycan structure 273:288 Among those applications, a specific one is to characterize glycan structure. 26800543 5 10 theme N-linked 735:742 arg1 glycopeptides 744:756 N-linked glycopeptides 735:756 N-linked glycopeptides 735:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 4 11 from identification 499:512 arg1 data 554:557 mass spectral data 540:557 mass spectral data 540:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 5 12 theme de 617:618 arg1 problem 636:642 the glycan de novo sequencing problem 606:642 the glycan de novo sequencing problem 606:642 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 5 13 theme glycopeptides 744:756 arg1 spectra 724:730 HCD MS/MS spectra 714:730 HCD MS/MS spectra of N-linked glycopeptides 714:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 5 14 theme sequencing 625:634 arg1 problem 636:642 the glycan de novo sequencing problem 606:642 the glycan de novo sequencing problem 606:642 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 7 15 theme MS/MS 988:992 arg1 spectra 994:1000 HCD MS/MS spectra 984:1000 HCD MS/MS spectra 984:1000 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 7 16 theme Experimental 881:892 arg1 results 894:900 Experimental results 881:900 Experimental results 881:900 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 5 17 dep de 690:691 arg1 novo 693:696 novo 693:696 novo 693:696 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 6 18 theme MS/MS 865:869 arg1 spectrum 871:878 MS/MS spectrum 865:878 MS/MS spectrum 865:878 The algorithm proceeds in a carefully designate pathway to construct the best matched tree structure from MS/MS spectrum. 26800543 5 19 link N-linked 735:742 arg1 glycopeptides 744:756 N-linked glycopeptides 735:756 N-linked glycopeptides 735:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 6 20 dep pathway 807:813 arg1 designate 797:805 designate 797:805 designate 797:805 The algorithm proceeds in a carefully designate pathway to construct the best matched tree structure from MS/MS spectrum. 26800543 0 21 theme Effective 3:11 arg1 Approach 13:20 An Effective Approach 0:20 An Effective Approach for Glycan Structure 0:41 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 1 22 theme Mass 80:83 arg1 spectrometry 85:96 Mass spectrometry 80:96 Mass spectrometry 80:96 Mass spectrometry has become a widely used analytical technique for proteomics study because of its high throughput and sensitivity. 26800543 7 23 theme HCD 984:986 arg1 spectra 994:1000 HCD MS/MS spectra 984:1000 HCD MS/MS spectra 984:1000 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 7 24 theme glycan 961:966 arg1 structures 968:977 glycan structures 961:977 glycan structures from HCD MS/MS spectra 961:1000 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 1 25 theme high 180:183 arg1 throughput 185:194 its high throughput 176:194 its high throughput 176:194 Mass spectrometry has become a widely used analytical technique for proteomics study because of its high throughput and sensitivity. 26800543 5 26 theme HCD 714:716 arg1 spectra 724:730 HCD MS/MS spectra 714:730 HCD MS/MS spectra of N-linked glycopeptides 714:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 0 27 from Spectra 71:77 arg1 Sequencing 51:60 De Novo Sequencing 43:60 De Novo Sequencing 43:60 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 5 28 gly glycopeptides 744:756 arg2 glycopeptides 744:756 N-linked glycopeptides 735:756 N-linked glycopeptides 735:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 4 29 from data 554:557 arg1 identification 499:512 the identification 495:512 the identification of glycan structures from mass spectral data 495:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 4 29 from data 554:557 arg1 structures 524:533 glycan structures 517:533 glycan structures from mass spectral data 517:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 5 30 theme MS/MS 718:722 arg1 spectra 724:730 HCD MS/MS spectra 714:730 HCD MS/MS spectra of N-linked glycopeptides 714:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 0 31 theme Glycan 26:31 arg1 Structure 33:41 Glycan Structure 26:41 Glycan Structure 26:41 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 4 32 theme mass 540:543 arg1 data 554:557 mass spectral data 540:557 mass spectral data 540:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 4 33 theme computational 461:473 arg1 methods 475:481 effective computational methods 451:481 effective computational methods 451:481 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 0 34 dep De 43:44 arg1 Novo 46:49 Novo 46:49 Novo 46:49 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 5 35 theme glycan 683:688 arg1 sequencing 698:707 glycan de novo sequencing 683:707 glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides 683:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 4 36 theme effective 451:459 arg1 methods 475:481 effective computational methods 451:481 effective computational methods 451:481 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 0 37 theme De 43:44 arg1 Sequencing 51:60 De Novo Sequencing 43:60 De Novo Sequencing 43:60 An Effective Approach for Glycan Structure De Novo Sequencing From HCD Spectra. 26800543 5 38 theme de 690:691 arg1 sequencing 698:707 glycan de novo sequencing 683:707 glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides 683:756 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 5 39 dep de 617:618 arg1 novo 620:623 novo 620:623 novo 620:623 In our research, we mathematically formulated the glycan de novo sequencing problem and proposed a heuristic algorithm for glycan de novo sequencing from HCD MS/MS spectra of N-linked glycopeptides. 26800543 4 40 theme spectral 545:552 arg1 data 554:557 mass spectral data 540:557 mass spectral data 540:557 Therefore, it is significant to develop effective computational methods to automate the identification of glycan structures from mass spectral data. 26800543 6 41 theme tree 845:848 arg1 structure 850:858 the best matched tree structure 828:858 the best matched tree structure from MS/MS spectrum 828:878 The algorithm proceeds in a carefully designate pathway to construct the best matched tree structure from MS/MS spectrum. 26800543 1 42 theme used 118:121 arg1 technique 134:142 a widely used analytical technique 109:142 a widely used analytical technique for proteomics study 109:163 Mass spectrometry has become a widely used analytical technique for proteomics study because of its high throughput and sensitivity. 26800543 7 43 theme proposed 918:925 arg1 approach 927:934 our proposed approach 914:934 our proposed approach 914:934 Experimental results showed that our proposed approach can effectively identify glycan structures from HCD MS/MS spectra. 26800543 3 44 theme occurred 321:328 arg1 Glycosylation 291:303 Glycosylation 291:303 Glycosylation 291:303 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 44 theme occurred 321:328 arg1 relevant 383:390 relevant 383:390 relevant 383:390 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 44 theme occurred 321:328 arg1 modification 349:360 a frequently occurred post-translational modification 308:360 a frequently occurred post-translational modification of proteins which is relevant to humans' health 308:408 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 6 45 theme matched 837:843 arg1 structure 850:858 the best matched tree structure 828:858 the best matched tree structure from MS/MS spectrum 828:878 The algorithm proceeds in a carefully designate pathway to construct the best matched tree structure from MS/MS spectrum. 26800543 1 46 theme analytical 123:132 arg1 technique 134:142 a widely used analytical technique 109:142 a widely used analytical technique for proteomics study 109:163 Mass spectrometry has become a widely used analytical technique for proteomics study because of its high throughput and sensitivity. 26800543 3 47 theme post-translational 330:347 arg1 Glycosylation 291:303 Glycosylation 291:303 Glycosylation 291:303 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 47 theme post-translational 330:347 arg1 relevant 383:390 relevant 383:390 relevant 383:390 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 3 47 theme post-translational 330:347 arg1 modification 349:360 a frequently occurred post-translational modification 308:360 a frequently occurred post-translational modification of proteins which is relevant to humans' health 308:408 Glycosylation is a frequently occurred post-translational modification of proteins which is relevant to humans' health. 26800543 6 48 from spectrum 871:878 arg1 structure 850:858 the best matched tree structure 828:858 the best matched tree structure from MS/MS spectrum 828:878 The algorithm proceeds in a carefully designate pathway to construct the best matched tree structure from MS/MS spectrum. 26330553 6 0 theme synthesis 988:996 arg1 genes 998:1002 the synthesis genes 984:1002 the synthesis genes 984:1002 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 1 theme O16 1143:1145 arg1 antigen 1147:1153 the original O16 antigen 1130:1153 the original O16 antigen 1130:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 2 theme O-antigen 901:909 arg1 production 911:920 O-antigen production 901:920 O-antigen production 901:920 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 10 3 theme genes 1813:1817 arg1 deletion 1783:1790 deletion 1783:1790 deletion of the glucosylation genes from the E. coli chromosome 1783:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 1 4 theme bacteriophage 307:319 arg1 receptor 321:328 the bacteriophage receptor 303:328 the bacteriophage receptor 303:328 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 10 5 from deletion 1783:1790 arg1 chromosome 1836:1845 the E. coli chromosome 1824:1845 the E. coli chromosome 1824:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 7 6 theme terrigena 1167:1175 arg1 ATCC 1177:1180 Raoultella terrigena ATCC 33257 1156:1186 Raoultella terrigena ATCC 33257 1156:1186 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 7 7 contain possessing 1210:1219 arg1 O-antigen 1200:1208 an O-antigen 1197:1208 an O-antigen possessing the same disaccharide motif 1197:1247 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 7 7 contain possessing 1210:1219 arg2 motif 1243:1247 the same disaccharide motif 1221:1247 the same disaccharide motif 1221:1247 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 11 8 theme O-antigen 1988:1996 arg1 glucosylation 1998:2010 bacteriophage-mediated O-antigen glucosylation 1965:2010 bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways 1965:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 11 9 theme transporter-dependent 2019:2039 arg1 pathways 2041:2048 ABC transporter-dependent pathways 2015:2048 ABC transporter-dependent pathways 2015:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 10 10 theme coli 1831:1834 arg1 chromosome 1836:1845 the E. coli chromosome 1824:1845 the E. coli chromosome 1824:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 11 theme ABC 1697:1699 arg1 transporter 1701:1711 a functional ABC transporter 1684:1711 a functional ABC transporter 1684:1711 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 9 12 theme Structural 1543:1552 arg1 determination 1554:1566 Structural determination 1543:1566 Structural determination using NMR 1543:1576 Structural determination using NMR revealed the addition of glucose side chains to the repeat units. 26330553 4 13 theme synthesis 703:711 arg1 systems 713:719 the two most prevalent O-antigen synthesis systems 670:719 the two most prevalent O-antigen synthesis systems 670:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 6 14 from K-12 953:956 arg1 cryptic 925:931 cryptic 925:931 cryptic 925:931 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 5 15 theme glucosylation 775:787 arg1 potential 789:797 the glucosylation potential 771:797 the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system 771:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 10 16 theme native 1858:1863 arg1 antisera 1871:1878 native level antisera sensitivity and structure 1858:1904 native level antisera sensitivity and structure 1858:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 16 theme native 1858:1863 arg1 structure 1896:1904 structure 1896:1904 structure 1896:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 16 theme native 1858:1863 arg1 sensitivity 1880:1890 sensitivity 1880:1890 sensitivity 1880:1890 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 1 17 theme O-antigen 258:266 arg1 glycans 268:274 lipopolysaccharide O-antigen glycans 239:274 lipopolysaccharide O-antigen glycans 239:274 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 6 18 theme prophage 1020:1027 arg1 cluster 1043:1049 a prophage glucosylation cluster 1018:1049 a prophage glucosylation cluster 1018:1049 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 8 19 theme chemical 1523:1530 arg1 structure 1532:1540 an altered chemical structure 1512:1540 an altered chemical structure 1512:1540 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 2 20 theme vaccine 444:450 arg1 development 452:462 vaccine development 444:462 vaccine development 444:462 This can enhance virulence and has implications for antigenic diversity and vaccine development. 26330553 8 21 theme terrigena 1334:1342 arg1 biosynthesis 1354:1365 the R. terrigena O-antigen biosynthesis 1327:1365 the R. terrigena O-antigen biosynthesis genes 1327:1371 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 8 22 theme biosynthesis 1354:1365 arg1 genes 1367:1371 the R. terrigena O-antigen biosynthesis genes 1327:1371 the R. terrigena O-antigen biosynthesis genes 1327:1371 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 6 23 from cryptic 925:931 arg1 K-12 953:956 Escherichia coli K-12 936:956 Escherichia coli K-12 936:956 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 10 24 from chromosome 1836:1845 arg1 deletion 1783:1790 deletion 1783:1790 deletion of the glucosylation genes from the E. coli chromosome 1783:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 24 from chromosome 1836:1845 arg1 genes 1813:1817 the glucosylation genes 1795:1817 the glucosylation genes from the E. coli chromosome 1795:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 8 25 theme R. 1467:1468 arg1 structure 1487:1495 the native R. terrigena repeat structure 1456:1495 the native R. terrigena repeat structure 1456:1495 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 9 26 theme glucose 1603:1609 arg1 chains 1616:1621 glucose side chains 1603:1621 glucose side chains 1603:1621 Structural determination using NMR revealed the addition of glucose side chains to the repeat units. 26330553 7 27 theme transporter-dependent 1279:1299 arg1 system 1301:1306 an ABC transporter-dependent system 1272:1306 an ABC transporter-dependent system 1272:1306 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 9 28 theme chains 1616:1621 arg1 addition 1591:1598 the addition 1587:1598 the addition of glucose side chains to the repeat units 1587:1641 Structural determination using NMR revealed the addition of glucose side chains to the repeat units. 26330553 8 29 theme reduced 1406:1412 arg1 reactivity 1414:1423 reduced reactivity 1406:1423 reduced reactivity 1406:1423 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 0 30 theme Cassette 108:115 arg1 Mechanism 154:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 2 31 contain has 399:401 arg1 This 368:371 This 368:371 This 368:371 This can enhance virulence and has implications for antigenic diversity and vaccine development. 26330553 2 31 contain has 399:401 arg2 implications 403:414 implications 403:414 implications for antigenic diversity and vaccine development 403:462 This can enhance virulence and has implications for antigenic diversity and vaccine development. 26330553 11 32 from glucosylation 1998:2010 arg1 pathways 2041:2048 ABC transporter-dependent pathways 2015:2048 ABC transporter-dependent pathways 2015:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 1 33 dep encode 194:199 arg1 resulting 334:342 resulting 334:342 resulting in serotype conversion 334:365 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 1 33 dep encode 194:199 arg1 altering 277:284 altering 277:284 altering the structure of the bacteriophage receptor 277:328 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 5 34 theme model 804:808 arg1 O-antigen 810:818 a model O-antigen 802:818 a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system 802:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 0 35 theme Transporter-dependent 123:143 arg1 Mechanism 154:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 3 36 located found 524:528 arg2 glucosylation 476:488 Side chain glucosylation 465:488 Side chain glucosylation 465:488 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 36 located found 524:528 arg1 number 535:540 a number 533:540 a number of bacterial species 533:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 36 located found 524:528 arg2 strategy 515:522 a common modification strategy 493:522 a common modification strategy found in a number of bacterial species 493:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 6 37 located found 1121:1125 arg1 antigen 1147:1153 the original O16 antigen 1130:1153 the original O16 antigen 1130:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 37 located found 1121:1125 arg2 motif 1115:1119 an α-l-Rha-(1→3)-d-GlcNAc motif 1089:1119 an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen 1089:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 0 38 theme ATP-binding 96:106 arg1 Cassette 108:115 ATP-binding Cassette 96:115 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 0 38 theme ATP-binding 96:106 arg1 ABC 118:120 ABC 118:120 ABC 118:120 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 3 39 theme common 495:500 arg1 strategy 515:522 a common modification strategy 493:522 a common modification strategy found in a number of bacterial species 493:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 39 theme common 495:500 arg1 glucosylation 476:488 Side chain glucosylation 465:488 Side chain glucosylation 465:488 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 0 40 theme Bacteriophage-mediated 0:21 arg1 Glucosylation 23:35 Bacteriophage-mediated Glucosylation 0:35 Bacteriophage-mediated Glucosylation 0:35 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 4 41 located observed 601:608 arg2 glucosylation 573:585 glucosylation 573:585 glucosylation 573:585 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 4 41 located observed 601:608 arg1 O-antigens 613:622 O-antigens 613:622 O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems 613:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 7 42 theme same 1225:1228 arg1 motif 1243:1247 the same disaccharide motif 1221:1247 the same disaccharide motif 1221:1247 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 1 43 theme serotype 347:354 arg1 conversion 356:365 serotype conversion 347:365 serotype conversion 347:365 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 8 44 theme repeat 1480:1485 arg1 structure 1487:1495 the native R. terrigena repeat structure 1456:1495 the native R. terrigena repeat structure 1456:1495 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 0 45 theme Lipopolysaccharide 48:65 arg1 O-Antigens 67:76 Lipopolysaccharide O-Antigens 48:76 Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 48:162 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 3 46 theme chain 470:474 arg1 strategy 515:522 a common modification strategy 493:522 a common modification strategy found in a number of bacterial species 493:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 46 theme chain 470:474 arg1 glucosylation 476:488 Side chain glucosylation 465:488 Side chain glucosylation 465:488 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 6 47 dep Escherichia 936:946 arg1 coli 948:951 coli 948:951 coli 948:951 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 10 48 theme glucosylation 1799:1811 arg1 genes 1813:1817 the glucosylation genes 1795:1817 the glucosylation genes from the E. coli chromosome 1795:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 5 49 theme transporter-dependent 862:882 arg1 system 884:889 an ATP-binding cassette (ABC) transporter-dependent system 832:889 an ATP-binding cassette (ABC) transporter-dependent system 832:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 3 50 theme bacterial 545:553 arg1 species 555:561 bacterial species 545:561 bacterial species 545:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 10 51 with consistent 1714:1723 arg1 modification 1730:1741 modification 1730:1741 modification in the periplasm 1730:1758 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 52 theme O-antigen 1644:1652 arg1 modification 1654:1665 O-antigen modification 1644:1665 O-antigen modification 1644:1665 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 8 53 theme altered 1515:1521 arg1 structure 1532:1540 an altered chemical structure 1512:1540 an altered chemical structure 1512:1540 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 10 54 theme E. 1828:1829 arg1 chromosome 1836:1845 the E. coli chromosome 1824:1845 the E. coli chromosome 1824:1845 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 2 55 theme antigenic 420:428 arg1 diversity 430:438 antigenic diversity 420:438 antigenic diversity 420:438 This can enhance virulence and has implications for antigenic diversity and vaccine development. 26330553 6 56 theme original 1134:1141 arg1 antigen 1147:1153 the original O16 antigen 1130:1153 the original O16 antigen 1130:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 7 57 theme Raoultella 1156:1165 arg1 ATCC 1177:1180 Raoultella terrigena ATCC 33257 1156:1186 Raoultella terrigena ATCC 33257 1156:1186 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 1 58 theme receptor 321:328 arg1 structure 290:298 the structure 286:298 the structure of the bacteriophage receptor 286:328 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 6 59 from mutation 972:979 arg1 genes 998:1002 the synthesis genes 984:1002 the synthesis genes 984:1002 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 10 60 from modification 1730:1741 arg1 periplasm 1750:1758 the periplasm 1746:1758 the periplasm 1746:1758 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 5 61 theme cassette 847:854 arg1 system 884:889 an ATP-binding cassette (ABC) transporter-dependent system 832:889 an ATP-binding cassette (ABC) transporter-dependent system 832:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 10 62 theme functional 1686:1695 arg1 transporter 1701:1711 a functional ABC transporter 1684:1711 a functional ABC transporter 1684:1711 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 63 theme level 1865:1869 arg1 antisera 1871:1878 native level antisera sensitivity and structure 1858:1904 native level antisera sensitivity and structure 1858:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 63 theme level 1865:1869 arg1 structure 1896:1904 structure 1896:1904 structure 1896:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 63 theme level 1865:1869 arg1 sensitivity 1880:1890 sensitivity 1880:1890 sensitivity 1880:1890 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 1 64 theme lipopolysaccharide 239:256 arg1 glycans 268:274 lipopolysaccharide O-antigen glycans 239:274 lipopolysaccharide O-antigen glycans 239:274 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 6 65 theme glucosylation 1029:1041 arg1 cluster 1043:1049 a prophage glucosylation cluster 1018:1049 a prophage glucosylation cluster 1018:1049 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 8 66 theme structure 1532:1540 arg1 indicative 1498:1507 indicative 1498:1507 indicative 1498:1507 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 4 67 theme O-antigen 693:701 arg1 systems 713:719 the two most prevalent O-antigen synthesis systems 670:719 the two most prevalent O-antigen synthesis systems 670:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 1 68 theme glycans 268:274 arg1 structures 225:234 the structures 221:234 the structures of lipopolysaccharide O-antigen glycans 221:274 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 4 69 theme systems 713:719 arg1 one 663:665 one 663:665 one 663:665 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 4 69 theme systems 713:719 arg1 systems 713:719 the two most prevalent O-antigen synthesis systems 670:719 the two most prevalent O-antigen synthesis systems 670:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 4 69 theme systems 713:719 arg1 pathways 653:660 Wzy-dependent pathways 639:660 Wzy-dependent pathways 639:660 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 10 70 theme consistent 1714:1723 arg1 transporter 1701:1711 a functional ABC transporter 1684:1711 a functional ABC transporter 1684:1711 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 11 71 theme intrinsic 1930:1938 arg1 barriers 1952:1959 no intrinsic mechanistic barriers 1927:1959 no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways 1927:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 5 72 theme heterologous 742:753 arg1 system 755:760 a heterologous system 740:760 a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system 740:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 8 73 theme O-antigen 1344:1352 arg1 biosynthesis 1354:1365 the R. terrigena O-antigen biosynthesis 1327:1365 the R. terrigena O-antigen biosynthesis genes 1327:1371 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 1 74 theme Lysogenic 165:173 arg1 bacteriophages 175:188 Lysogenic bacteriophages 165:188 Lysogenic bacteriophages 165:188 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 1 74 theme Lysogenic 165:173 arg1 enzymes 201:207 enzymes 201:207 enzymes that modify the structures of lipopolysaccharide O-antigen glycans 201:274 Lysogenic bacteriophages may encode enzymes that modify the structures of lipopolysaccharide O-antigen glycans, altering the structure of the bacteriophage receptor and resulting in serotype conversion. 26330553 10 75 dep antisera 1871:1878 arg1 antisera 1871:1878 native level antisera sensitivity and structure 1858:1904 native level antisera sensitivity and structure 1858:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 75 dep antisera 1871:1878 arg1 structure 1896:1904 structure 1896:1904 structure 1896:1904 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 10 75 dep antisera 1871:1878 arg1 sensitivity 1880:1890 sensitivity 1880:1890 sensitivity 1880:1890 O-antigen modification was dependent on a functional ABC transporter, consistent with modification in the periplasm, and was eliminated by deletion of the glucosylation genes from the E. coli chromosome, restoring native level antisera sensitivity and structure. 26330553 11 76 theme mechanistic 1940:1950 arg1 barriers 1952:1959 no intrinsic mechanistic barriers 1927:1959 no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways 1927:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 7 77 theme ABC 1275:1277 arg1 system 1301:1306 an ABC transporter-dependent system 1272:1306 an ABC transporter-dependent system 1272:1306 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 9 78 theme side 1611:1614 arg1 chains 1616:1621 glucose side chains 1603:1621 glucose side chains 1603:1621 Structural determination using NMR revealed the addition of glucose side chains to the repeat units. 26330553 5 79 theme ATP-binding 835:845 arg1 ABC 857:859 ABC 857:859 ABC 857:859 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 5 79 theme ATP-binding 835:845 arg1 cassette 847:854 an ATP-binding cassette 832:854 an ATP-binding cassette (ABC) transporter-dependent system 832:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 6 80 theme GlcNAc 1071:1076 arg1 residue 1078:1084 the GlcNAc residue 1067:1084 the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen 1067:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 81 from residue 1078:1084 arg1 motif 1115:1119 an α-l-Rha-(1→3)-d-GlcNAc motif 1089:1119 an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen 1089:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 9 82 theme repeat 1630:1635 arg1 units 1637:1641 the repeat units 1626:1641 the repeat units 1626:1641 Structural determination using NMR revealed the addition of glucose side chains to the repeat units. 26330553 0 83 theme Assembly 145:152 arg1 Mechanism 154:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism 93:162 Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism. 26330553 5 84 theme O-antigen 810:818 arg1 potential 789:797 the glucosylation potential 771:797 the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system 771:889 Here we exploited a heterologous system to study the glucosylation potential of a model O-antigen produced in an ATP-binding cassette (ABC) transporter-dependent system. 26330553 7 85 theme disaccharide 1230:1241 arg1 motif 1243:1247 the same disaccharide motif 1221:1247 the same disaccharide motif 1221:1247 Raoultella terrigena ATCC 33257 produces an O-antigen possessing the same disaccharide motif, but its assembly uses an ABC transporter-dependent system. 26330553 3 86 theme Side 465:468 arg1 strategy 515:522 a common modification strategy 493:522 a common modification strategy found in a number of bacterial species 493:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 86 theme Side 465:468 arg1 glucosylation 476:488 Side chain glucosylation 465:488 Side chain glucosylation 465:488 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 87 theme modification 502:513 arg1 strategy 515:522 a common modification strategy 493:522 a common modification strategy found in a number of bacterial species 493:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 3 87 theme modification 502:513 arg1 glucosylation 476:488 Side chain glucosylation 465:488 Side chain glucosylation 465:488 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 6 88 contain possesses 1008:1016 arg2 cluster 1043:1049 a prophage glucosylation cluster 1018:1049 a prophage glucosylation cluster 1018:1049 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 88 contain possesses 1008:1016 arg1 it 1005:1006 it 1005:1006 it 1005:1006 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 6 89 theme -d-GlcNAc 1105:1113 arg1 motif 1115:1119 an α-l-Rha-(1→3)-d-GlcNAc motif 1089:1119 an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen 1089:1153 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 4 90 theme Wzy-dependent 639:651 arg1 one 663:665 one 663:665 one 663:665 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 4 90 theme Wzy-dependent 639:651 arg1 systems 713:719 the two most prevalent O-antigen synthesis systems 670:719 the two most prevalent O-antigen synthesis systems 670:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 4 90 theme Wzy-dependent 639:651 arg1 pathways 653:660 Wzy-dependent pathways 639:660 Wzy-dependent pathways 639:660 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 8 91 theme native 1460:1465 arg1 structure 1487:1495 the native R. terrigena repeat structure 1456:1495 the native R. terrigena repeat structure 1456:1495 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 8 92 theme terrigena 1470:1478 arg1 structure 1487:1495 the native R. terrigena repeat structure 1456:1495 the native R. terrigena repeat structure 1456:1495 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 6 93 theme Escherichia 936:946 arg1 K-12 953:956 Escherichia coli K-12 936:956 Escherichia coli K-12 936:956 Although O-antigen production is cryptic in Escherichia coli K-12, because of a mutation in the synthesis genes, it possesses a prophage glucosylation cluster, which modifies the GlcNAc residue in an α-l-Rha-(1→3)-d-GlcNAc motif found in the original O16 antigen. 26330553 11 94 theme bacteriophage-mediated 1965:1986 arg1 glucosylation 1998:2010 bacteriophage-mediated O-antigen glucosylation 1965:2010 bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways 1965:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 26330553 8 95 theme indicative 1498:1507 arg1 reactivity 1414:1423 reduced reactivity 1406:1423 reduced reactivity 1406:1423 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 3 96 theme species 555:561 arg1 number 535:540 a number 533:540 a number of bacterial species 533:561 Side chain glucosylation is a common modification strategy found in a number of bacterial species. 26330553 8 97 theme R. 1331:1332 arg1 biosynthesis 1354:1365 the R. terrigena O-antigen biosynthesis 1327:1365 the R. terrigena O-antigen biosynthesis genes 1327:1371 E. coli harboring the R. terrigena O-antigen biosynthesis genes produced an O-antigen displaying reduced reactivity toward antisera raised against the native R. terrigena repeat structure, indicative of an altered chemical structure. 26330553 4 98 theme prevalent 683:691 arg1 systems 713:719 the two most prevalent O-antigen synthesis systems 670:719 the two most prevalent O-antigen synthesis systems 670:719 To date, glucosylation has only been observed in O-antigens synthesized by Wzy-dependent pathways, one of the two most prevalent O-antigen synthesis systems. 26330553 11 99 theme ABC 2015:2017 arg1 pathways 2041:2048 ABC transporter-dependent pathways 2015:2048 ABC transporter-dependent pathways 2015:2048 There are therefore no intrinsic mechanistic barriers for bacteriophage-mediated O-antigen glucosylation in ABC transporter-dependent pathways. 28915952 3 0 theme pathologic 474:483 arg1 features 485:492 pathologic features 474:492 pathologic features 474:492 The link between TNF-alpha blockers and IgAN may be hypothesized examining diseases which share pathologic features. 28915952 1 1 theme Factor 219:224 arg1 -alpha 231:236 anti-Tumor Necrosis Factor (TNF)-alpha 199:236 anti-Tumor Necrosis Factor (TNF)-alpha agents 199:243 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 4 2 from object 571:576 arg1 respect 503:509 this respect 498:509 this respect 498:509 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 1 3 dep -alpha 231:236 arg1 agents 238:243 agents 238:243 anti-Tumor Necrosis Factor (TNF)-alpha agents 199:243 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 4 4 gly glycosylation 631:643 arg1 molecules 653:661 IgA1 molecules 648:661 IgA1 molecules 648:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 2 5 from deposition 291:300 arg1 walls 347:351 vessel walls 340:351 vessel walls in the kidney mesangium 340:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 2 6 theme immune 320:325 arg1 complexes 327:335 IgA-containing immune complexes 305:335 IgA-containing immune complexes 305:335 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 1 7 theme TNF 227:229 arg1 -alpha 231:236 anti-Tumor Necrosis Factor (TNF)-alpha 199:236 anti-Tumor Necrosis Factor (TNF)-alpha agents 199:243 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 5 8 theme glycan 718:723 arg1 structures 725:734 glycan structures 718:734 glycan structures of TNF-alpha inhibitors 718:758 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 2 9 theme complexes 327:335 arg1 feature 278:284 The characteristic pathological feature 246:284 The characteristic pathological feature 246:284 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 2 9 theme complexes 327:335 arg1 deposition 291:300 a deposition 289:300 a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium 289:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 8 10 theme alpha 1189:1193 arg1 agents 1195:1200 anti-TNF alpha agents 1180:1200 anti-TNF alpha agents 1180:1200 Such a test would be useful also before administration of anti-TNF alpha agents. 28915952 7 11 theme blockers 1112:1119 arg1 introduction 1086:1097 introduction 1086:1097 introduction of TNF-alpha blockers 1086:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 2 12 theme kidney 360:365 arg1 mesangium 367:375 the kidney mesangium 356:375 the kidney mesangium 356:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 8 13 theme anti-TNF 1180:1187 arg1 agents 1195:1200 anti-TNF alpha agents 1180:1200 anti-TNF alpha agents 1180:1200 Such a test would be useful also before administration of anti-TNF alpha agents. 28915952 8 14 theme agents 1195:1200 arg1 administration 1162:1175 administration 1162:1175 administration of anti-TNF alpha agents 1162:1200 Such a test would be useful also before administration of anti-TNF alpha agents. 28915952 9 15 with treatment 1270:1278 arg1 blockers 1289:1296 TNF blockers 1285:1296 TNF blockers 1285:1296 The presence of aberrant IgA1 may represent a contraindication for treatment with TNF blockers. 28915952 4 16 theme aberrant 622:629 arg1 glycosylation 631:643 aberrant glycosylation 622:643 aberrant glycosylation of IgA1 molecules 622:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 9 17 theme aberrant 1219:1226 arg1 IgA1 1228:1231 aberrant IgA1 1219:1231 aberrant IgA1 1219:1231 The presence of aberrant IgA1 may represent a contraindication for treatment with TNF blockers. 28915952 4 18 theme IgA1 648:651 arg1 molecules 653:661 IgA1 molecules 648:661 IgA1 molecules 648:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 0 19 theme IgA 0:2 arg1 nephropathy 4:14 IgA nephropathy 0:14 IgA nephropathy during treatment with TNF-alpha blockers: 0:56 IgA nephropathy during treatment with TNF-alpha blockers: Could it be predicted? 28915952 7 20 theme TNF-alpha 1102:1110 arg1 blockers 1112:1119 TNF-alpha blockers 1102:1119 TNF-alpha blockers 1102:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 9 21 theme IgA1 1228:1231 arg1 presence 1207:1214 The presence 1203:1214 The presence of aberrant IgA1 1203:1231 The presence of aberrant IgA1 may represent a contraindication for treatment with TNF blockers. 28915952 1 22 theme Immunoglobulin 81:94 arg1 IgA 99:101 IgA 99:101 IgA 99:101 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 1 22 theme Immunoglobulin 81:94 arg1 A 96:96 Immunoglobulin A 81:96 Immunoglobulin A (IgA) nephropathy (IgAN) 81:121 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 3 23 theme TNF-alpha 395:403 arg1 blockers 405:412 TNF-alpha blockers 395:412 TNF-alpha blockers 395:412 The link between TNF-alpha blockers and IgAN may be hypothesized examining diseases which share pathologic features. 28915952 4 24 theme IgAN 523:526 arg1 Purpura 549:555 IgAN and Henoch Schonlein Purpura 523:555 Purpura 549:555 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 24 theme IgAN 523:526 arg1 object 571:576 the object 567:576 the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules 567:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 2 25 theme pathological 265:276 arg1 feature 278:284 The characteristic pathological feature 246:284 The characteristic pathological feature 246:284 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 2 25 theme pathological 265:276 arg1 deposition 291:300 a deposition 289:300 a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium 289:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 7 26 theme Such 964:967 arg1 hypothesis 969:978 Such hypothesis 964:978 Such hypothesis 964:978 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 4 27 theme pathogenetic 601:612 arg1 role 614:617 a pathogenetic role 599:617 a pathogenetic role of aberrant glycosylation of IgA1 molecules 599:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 5 28 theme large 815:819 arg1 complexes 838:846 large antigen-antibody complexes 815:846 large antigen-antibody complexes 815:846 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 4 29 theme Schonlein 539:547 arg1 Purpura 549:555 IgAN and Henoch Schonlein Purpura 523:555 Purpura 549:555 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 29 theme Schonlein 539:547 arg1 object 571:576 the object 567:576 the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules 567:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 2 30 theme characteristic 250:263 arg1 feature 278:284 The characteristic pathological feature 246:284 The characteristic pathological feature 246:284 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 2 30 theme characteristic 250:263 arg1 deposition 291:300 a deposition 289:300 a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium 289:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 1 31 theme pharmacological 163:177 arg1 agents 179:184 pharmacological agents 163:184 pharmacological agents 163:184 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 0 32 theme TNF-alpha 38:46 arg1 blockers 48:55 TNF-alpha blockers 38:55 TNF-alpha blockers 38:55 IgA nephropathy during treatment with TNF-alpha blockers: Could it be predicted? 28915952 4 33 theme Henoch 532:537 arg1 Purpura 549:555 IgAN and Henoch Schonlein Purpura 523:555 Purpura 549:555 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 33 theme Henoch 532:537 arg1 object 571:576 the object 567:576 the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules 567:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 5 34 theme anti-drug 689:697 arg1 antibodies 699:708 anti-drug antibodies 689:708 anti-drug antibodies against glycan structures of TNF-alpha inhibitors 689:758 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 6 35 theme IgA 871:873 arg1 complexes 875:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes are then able to deposit in the mesangium and activate the complement cascade. 28915952 4 36 theme glycosylation 631:643 arg1 role 614:617 a pathogenetic role 599:617 a pathogenetic role of aberrant glycosylation of IgA1 molecules 599:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 6 37 theme complement 944:953 arg1 cascade 955:961 the complement cascade 940:961 the complement cascade 940:961 These large polymeric IgA complexes are then able to deposit in the mesangium and activate the complement cascade. 28915952 6 38 theme polymeric 861:869 arg1 complexes 875:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes are then able to deposit in the mesangium and activate the complement cascade. 28915952 0 39 with treatment 23:31 arg1 blockers 48:55 TNF-alpha blockers 38:55 TNF-alpha blockers 38:55 IgA nephropathy during treatment with TNF-alpha blockers: Could it be predicted? 28915952 5 40 theme serum 784:788 arg1 IgA1 799:802 serum aberrant IgA1 784:802 serum aberrant IgA1 leading to large antigen-antibody complexes 784:846 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 6 41 theme large 855:859 arg1 complexes 875:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes 849:883 These large polymeric IgA complexes are then able to deposit in the mesangium and activate the complement cascade. 28915952 2 42 from walls 347:351 arg1 mesangium 367:375 the kidney mesangium 356:375 the kidney mesangium 356:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 2 43 theme IgA-containing 305:318 arg1 complexes 327:335 IgA-containing immune complexes 305:335 IgA-containing immune complexes 305:335 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 5 44 theme aberrant 790:797 arg1 IgA1 799:802 serum aberrant IgA1 784:802 serum aberrant IgA1 leading to large antigen-antibody complexes 784:846 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 4 45 theme idiopathic 512:521 arg1 Purpura 549:555 IgAN and Henoch Schonlein Purpura 523:555 Purpura 549:555 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 45 theme idiopathic 512:521 arg1 object 571:576 the object 567:576 the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules 567:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 7 46 theme patients 1051:1058 arg1 IgA1 1043:1046 galactose-deficient IgA1 1023:1046 galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers 1023:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 7 47 theme galactose-deficient 1023:1041 arg1 IgA1 1043:1046 galactose-deficient IgA1 1023:1046 galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers 1023:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 7 48 theme serum 1007:1011 arg1 levels 1013:1018 serum levels 1007:1018 serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers 1007:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 1 49 theme A 96:96 arg1 nephropathy 104:114 Immunoglobulin A (IgA) nephropathy 81:114 Immunoglobulin A (IgA) nephropathy (IgAN) 81:121 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 1 49 theme A 96:96 arg1 IgAN 117:120 IgAN 117:120 IgAN 117:120 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 7 50 theme IgA1 1043:1046 arg1 levels 1013:1018 serum levels 1007:1018 serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers 1007:1119 Such hypothesis may be tested by measuring serum levels of galactose-deficient IgA1 of patients developing IgAN following introduction of TNF-alpha blockers. 28915952 9 51 theme TNF 1285:1287 arg1 blockers 1289:1296 TNF blockers 1285:1296 TNF blockers 1285:1296 The presence of aberrant IgA1 may represent a contraindication for treatment with TNF blockers. 28915952 5 52 theme TNF-alpha 739:747 arg1 inhibitors 749:758 TNF-alpha inhibitors 739:758 TNF-alpha inhibitors 739:758 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 5 53 theme antigen-antibody 821:836 arg1 complexes 838:846 large antigen-antibody complexes 815:846 large antigen-antibody complexes 815:846 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 2 54 theme vessel 340:345 arg1 walls 347:351 vessel walls 340:351 vessel walls in the kidney mesangium 340:375 The characteristic pathological feature is a deposition of IgA-containing immune complexes in vessel walls in the kidney mesangium. 28915952 1 55 theme anti-Tumor 199:208 arg1 -alpha 231:236 anti-Tumor Necrosis Factor (TNF)-alpha 199:236 anti-Tumor Necrosis Factor (TNF)-alpha agents 199:243 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 28915952 5 56 theme inhibitors 749:758 arg1 structures 725:734 glycan structures 718:734 glycan structures of TNF-alpha inhibitors 718:758 The Authors suggest that anti-drug antibodies against glycan structures of TNF-alpha inhibitors may cross react against serum aberrant IgA1 leading to large antigen-antibody complexes. 28915952 4 57 theme studies 581:587 arg1 Purpura 549:555 IgAN and Henoch Schonlein Purpura 523:555 Purpura 549:555 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 57 theme studies 581:587 arg1 object 571:576 the object 567:576 the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules 567:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 4 58 theme molecules 653:661 arg1 glycosylation 631:643 aberrant glycosylation 622:643 aberrant glycosylation of IgA1 molecules 622:661 In this respect, idiopathic IgAN and Henoch Schonlein Purpura have been the object of studies revealing a pathogenetic role of aberrant glycosylation of IgA1 molecules. 28915952 1 59 theme Necrosis 210:217 arg1 -alpha 231:236 anti-Tumor Necrosis Factor (TNF)-alpha 199:236 anti-Tumor Necrosis Factor (TNF)-alpha agents 199:243 Immunoglobulin A (IgA) nephropathy (IgAN) may sometimes be related to exposure to pharmacological agents, among which anti-Tumor Necrosis Factor (TNF)-alpha agents. 27780868 0 0 theme Active 81:86 arg1 Site 88:91 Active Site 81:91 Active Site of Cellobiohydrolase Cel7A 81:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 0 0 theme Active 81:86 arg1 Cel7A 114:118 Cellobiohydrolase Cel7A 96:118 Cellobiohydrolase Cel7A 96:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 5 1 theme them 729:732 arg1 Trp-38 746:751 Trp-38 746:751 Trp-38 746:751 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 1 theme them 729:732 arg1 Trp-40 735:740 Trp-40 735:740 Trp-40 735:740 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 1 theme them 729:732 arg1 them 729:732 them 729:732 them 729:732 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 1 theme them 729:732 arg1 two 722:724 two 722:724 two 722:724 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 9 2 dep substitution 1240:1251 arg1 to 1233:1234 to 1233:1234 to 1233:1234 The Trp-38 to Ala substitution resulted in increased off-rates and decreased processivity. 27780868 3 3 theme carbohydrate-binding 485:504 arg1 module 506:511 a smaller carbohydrate-binding module 475:511 a smaller carbohydrate-binding module (CBM) 475:517 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 3 3 theme carbohydrate-binding 485:504 arg1 CBM 514:516 CBM 514:516 CBM 514:516 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 4 4 theme tunnel-shaped 572:584 arg1 site 593:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 10 5 theme CBM-linker 1416:1425 arg1 presence 1400:1407 the presence 1396:1407 the presence of the CBM-linker 1396:1425 The effect of the Trp-38 to Ala substitution on on-rates was strongly dependent on the presence of the CBM-linker. 27780868 2 6 theme reesei 362:367 arg1 Cel7A 341:345 cellobiohydrolase Cel7A 323:345 cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) 323:377 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 1 7 theme green 262:266 arg1 chemistry 268:276 green chemistry 262:276 green chemistry 262:276 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 5 8 theme Trp 704:706 arg1 residues 708:715 four Trp residues 699:715 four Trp residues with two of them, Trp-40 and Trp-38, 699:752 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 9 9 theme Trp-38 1226:1231 arg1 substitution 1240:1251 The Trp-38 to Ala substitution 1222:1251 The Trp-38 to Ala substitution 1222:1251 The Trp-38 to Ala substitution resulted in increased off-rates and decreased processivity. 27780868 7 10 theme TrCel7A 1038:1044 arg1 Trp-38 1028:1033 Trp-38 1028:1033 Trp-38 1028:1033 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 7 10 theme TrCel7A 1038:1044 arg1 CBM-linker 1013:1022 CBM-linker 1013:1022 CBM-linker 1013:1022 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 3 11 gly glycosylated 541:552 arg1 peptide 561:567 the glycosylated linker peptide 537:567 the glycosylated linker peptide 537:567 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 0 12 theme Cel7A 114:118 arg1 Site 88:91 Active Site 81:91 Active Site of Cellobiohydrolase Cel7A 81:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 0 12 theme Cel7A 114:118 arg1 Cel7A 114:118 Cellobiohydrolase Cel7A 96:118 Cellobiohydrolase Cel7A 96:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 4 13 theme glucose 630:636 arg1 sites 651:655 10 glucose unit binding sites 627:655 10 glucose unit binding sites 627:655 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 4 14 theme active 586:591 arg1 site 593:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 2 15 theme glycoside 290:298 arg1 hydrolases 300:309 Processive glycoside hydrolases 279:309 Processive glycoside hydrolases (GHs) 279:315 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 15 theme glycoside 290:298 arg1 components 387:396 key components 383:396 key components of efficient enzyme systems 383:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 15 theme glycoside 290:298 arg1 GHs 312:314 GHs 312:314 GHs 312:314 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 8 16 theme on-rate 1176:1182 arg1 affinity 1198:1205 the on-rate and substrate affinity 1172:1205 the on-rate and substrate affinity of the enzyme 1172:1219 The CBM-linker increased the on-rate and substrate affinity of the enzyme. 27780868 4 17 theme binding 643:649 arg1 sites 651:655 10 glucose unit binding sites 627:655 10 glucose unit binding sites 627:655 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 0 18 theme Cellobiohydrolase 96:112 arg1 Cel7A 114:118 Cellobiohydrolase Cel7A 96:118 Cellobiohydrolase Cel7A 96:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 6 19 theme numerous 833:840 arg1 studies 842:848 numerous studies 833:848 numerous studies 833:848 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 6 20 theme complex 934:940 arg1 mechanism 952:960 a complex multistep mechanism 932:960 a complex multistep mechanism of processive GHs 932:978 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 12 21 theme endoglucanase 1682:1694 arg1 presence 1670:1677 the presence 1666:1677 the presence of endoglucanase 1666:1694 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 2 22 theme Processive 279:288 arg1 hydrolases 300:309 Processive glycoside hydrolases 279:309 Processive glycoside hydrolases (GHs) 279:315 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 22 theme Processive 279:288 arg1 components 387:396 key components 383:396 key components of efficient enzyme systems 383:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 22 theme Processive 279:288 arg1 GHs 312:314 GHs 312:314 GHs 312:314 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 4 23 theme unit 638:641 arg1 sites 651:655 10 glucose unit binding sites 627:655 10 glucose unit binding sites 627:655 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 6 24 theme site 896:899 arg1 aromatics 901:909 CBM and active site aromatics 881:909 CBM and active site aromatics 881:909 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 10 25 theme Ala 1341:1343 arg1 substitution 1345:1356 Ala substitution 1341:1356 Ala substitution 1341:1356 The effect of the Trp-38 to Ala substitution on on-rates was strongly dependent on the presence of the CBM-linker. 27780868 7 26 with on- 1080:1082 arg1 endoglucanase 1132:1144 endoglucanase 1132:1144 endoglucanase 1132:1144 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 5 27 theme active 662:667 arg1 site 669:672 The active site 658:672 The active site of TrCel7A 658:683 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 27 theme active 662:667 arg1 TrCel7A 677:683 TrCel7A 677:683 TrCel7A 677:683 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 2 28 theme key 383:385 arg1 components 387:396 key components 383:396 key components of efficient enzyme systems 383:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 28 theme key 383:385 arg1 hydrolases 300:309 Processive glycoside hydrolases 279:309 Processive glycoside hydrolases (GHs) 279:315 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 0 29 theme Inter-domain 0:11 arg1 Synergism 13:21 Inter-domain Synergism 0:21 Inter-domain Synergism 0:21 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 7 30 theme CBM-linker 1013:1022 arg1 role 1001:1004 the role 997:1004 the role of the CBM-linker and Trp-38 of TrCel7A 997:1044 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 1 31 theme enzymatic 197:205 arg1 route 253:257 an important route 240:257 an important route in green chemistry 240:276 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 1 31 theme enzymatic 197:205 arg1 degradation 207:217 their enzymatic degradation 191:217 their enzymatic degradation to soluble sugars 191:235 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 5 32 theme substrate 761:769 arg1 sites 779:783 the substrate binding sites 757:783 the substrate binding sites near the tunnel entrance 757:808 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 33 theme TrCel7A 677:683 arg1 site 669:672 The active site 658:672 The active site of TrCel7A 658:683 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 33 theme TrCel7A 677:683 arg1 TrCel7A 677:683 TrCel7A 677:683 TrCel7A 677:683 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 7 34 with synergism 1117:1125 arg1 endoglucanase 1132:1144 endoglucanase 1132:1144 endoglucanase 1132:1144 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 3 35 theme glycosylated 541:552 arg1 peptide 561:567 the glycosylated linker peptide 537:567 the glycosylated linker peptide 537:567 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 7 36 with processivity 1099:1110 arg1 endoglucanase 1132:1144 endoglucanase 1132:1144 endoglucanase 1132:1144 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 8 37 theme substrate 1188:1196 arg1 affinity 1198:1205 the on-rate and substrate affinity 1172:1205 the on-rate and substrate affinity of the enzyme 1172:1219 The CBM-linker increased the on-rate and substrate affinity of the enzyme. 27780868 5 38 theme binding 771:777 arg1 sites 779:783 the substrate binding sites 757:783 the substrate binding sites near the tunnel entrance 757:808 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 6 39 theme aromatics 901:909 arg1 role 873:876 the role 869:876 the role of CBM and active site aromatics 869:909 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 7 40 with affinity 1070:1077 arg1 endoglucanase 1132:1144 endoglucanase 1132:1144 endoglucanase 1132:1144 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 3 41 theme linker 554:559 arg1 peptide 561:567 the glycosylated linker peptide 537:567 the glycosylated linker peptide 537:567 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 12 42 theme inter-domain 1579:1590 arg1 pre-requisite 1606:1618 pre-requisite 1606:1618 pre-requisite 1606:1618 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 12 42 theme inter-domain 1579:1590 arg1 synergism 1592:1600 The inter-domain synergism 1575:1600 The inter-domain synergism 1575:1600 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 2 43 theme systems 418:424 arg1 components 387:396 key components 383:396 key components of efficient enzyme systems 383:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 2 43 theme systems 418:424 arg1 hydrolases 300:309 Processive glycoside hydrolases 279:309 Processive glycoside hydrolases (GHs) 279:315 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 0 44 theme Efficient 39:47 arg1 Feeding 49:55 Efficient Feeding 39:55 Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A 39:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 3 45 theme smaller 477:483 arg1 module 506:511 a smaller carbohydrate-binding module 475:511 a smaller carbohydrate-binding module (CBM) 475:517 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 3 45 theme smaller 477:483 arg1 CBM 514:516 CBM 514:516 CBM 514:516 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 1 46 theme soluble 222:228 arg1 sugars 230:235 soluble sugars 222:235 soluble sugars 222:235 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 5 47 with residues 708:715 arg1 Trp-38 746:751 Trp-38 746:751 Trp-38 746:751 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 47 with residues 708:715 arg1 Trp-40 735:740 Trp-40 735:740 Trp-40 735:740 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 47 with residues 708:715 arg1 them 729:732 them 729:732 them 729:732 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 5 47 with residues 708:715 arg1 two 722:724 two 722:724 two 722:724 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 2 48 theme enzyme 411:416 arg1 systems 418:424 efficient enzyme systems 401:424 efficient enzyme systems 401:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 7 49 dep CBM-linker 1013:1022 arg1 the 1009:1011 the 1009:1011 the 1009:1011 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 6 50 theme active 889:894 arg1 site 896:899 active site 889:899 active site 889:899 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 2 51 theme efficient 401:409 arg1 systems 418:424 efficient enzyme systems 401:424 efficient enzyme systems 401:424 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 9 52 theme decreased 1289:1297 arg1 processivity 1299:1310 decreased processivity 1289:1310 decreased processivity 1289:1310 The Trp-38 to Ala substitution resulted in increased off-rates and decreased processivity. 27780868 6 53 theme GHs 976:978 arg1 mechanism 952:960 a complex multistep mechanism 932:960 a complex multistep mechanism of processive GHs 932:978 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 4 54 contain contains 618:625 arg1 site 593:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site 570:596 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 4 54 contain contains 618:625 arg2 sites 651:655 10 glucose unit binding sites 627:655 10 glucose unit binding sites 627:655 A tunnel-shaped active site rests in the CD and contains 10 glucose unit binding sites. 27780868 9 55 theme Ala 1236:1238 arg1 substitution 1240:1251 The Trp-38 to Ala substitution 1222:1251 The Trp-38 to Ala substitution 1222:1251 The Trp-38 to Ala substitution resulted in increased off-rates and decreased processivity. 27780868 8 56 theme enzyme 1214:1219 arg1 affinity 1198:1205 the on-rate and substrate affinity 1172:1205 the on-rate and substrate affinity of the enzyme 1172:1219 The CBM-linker increased the on-rate and substrate affinity of the enzyme. 27780868 7 57 theme binding 1062:1068 arg1 affinity 1070:1077 binding affinity 1062:1077 binding affinity 1062:1077 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 6 58 theme CBM 881:883 arg1 aromatics 901:909 CBM and active site aromatics 881:909 CBM and active site aromatics 881:909 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 1 59 from route 253:257 arg1 chemistry 268:276 green chemistry 262:276 green chemistry 262:276 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 7 60 theme Trp-38 1028:1033 arg1 role 1001:1004 the role 997:1004 the role of the CBM-linker and Trp-38 of TrCel7A 997:1044 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 10 61 from effect 1317:1322 arg1 on-rates 1361:1368 on-rates 1361:1368 on-rates 1361:1368 The effect of the Trp-38 to Ala substitution on on-rates was strongly dependent on the presence of the CBM-linker. 27780868 1 62 theme Structural 121:130 arg1 polysaccharides 132:146 Structural polysaccharides 121:146 Structural polysaccharides like cellulose and chitin 121:172 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 10 63 theme Trp-38 1331:1336 arg1 effect 1317:1322 The effect 1313:1322 The effect of the Trp-38 to Ala substitution on on-rates 1313:1368 The effect of the Trp-38 to Ala substitution on on-rates was strongly dependent on the presence of the CBM-linker. 27780868 10 63 theme Trp-38 1331:1336 arg1 dependent 1383:1391 dependent 1383:1391 dependent 1383:1391 The effect of the Trp-38 to Ala substitution on on-rates was strongly dependent on the presence of the CBM-linker. 27780868 9 64 theme increased 1265:1273 arg1 off-rates 1275:1283 increased off-rates 1265:1283 increased off-rates 1265:1283 The Trp-38 to Ala substitution resulted in increased off-rates and decreased processivity. 27780868 7 65 with off-rates 1088:1096 arg1 endoglucanase 1132:1144 endoglucanase 1132:1144 endoglucanase 1132:1144 Here we studied the role of the CBM-linker and Trp-38 of TrCel7A with respect to binding affinity, on- and off-rates, processivity, and synergism with endoglucanase. 27780868 6 66 theme processive 965:974 arg1 GHs 976:978 processive GHs 965:978 processive GHs 965:978 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 12 67 from degradation 1638:1648 arg1 presence 1670:1677 the presence 1666:1677 the presence of endoglucanase 1666:1694 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 2 68 theme cellobiohydrolase 323:339 arg1 Cel7A 341:345 cellobiohydrolase Cel7A 323:345 cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) 323:377 Processive glycoside hydrolases (GHs), like cellobiohydrolase Cel7A of Trichoderma reesei (TrCel7A) are key components of efficient enzyme systems. 27780868 11 69 theme cellulose 1537:1545 arg1 chain 1547:1551 the cellulose chain 1533:1551 the cellulose chain into the active site 1533:1572 This compensation between CBM-linker and Trp-38 indicates synergism between CBM-linker and CD in feeding the cellulose chain into the active site. 27780868 0 70 theme Chain 70:74 arg1 Feeding 49:55 Efficient Feeding 39:55 Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A 39:118 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 5 71 theme tunnel 794:799 arg1 entrance 801:808 the tunnel entrance 790:808 the tunnel entrance 790:808 The active site of TrCel7A is lined with four Trp residues with two of them, Trp-40 and Trp-38, in the substrate binding sites near the tunnel entrance. 27780868 3 72 theme catalytic 449:457 arg1 domain 459:464 a catalytic domain 447:464 a catalytic domain (CD) 447:469 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 3 72 theme catalytic 449:457 arg1 CD 467:468 CD 467:468 CD 467:468 TrCel7A consists of a catalytic domain (CD) and a smaller carbohydrate-binding module (CBM) connected through the glycosylated linker peptide. 27780868 11 73 theme active 1562:1567 arg1 site 1569:1572 the active site 1558:1572 the active site 1558:1572 This compensation between CBM-linker and Trp-38 indicates synergism between CBM-linker and CD in feeding the cellulose chain into the active site. 27780868 12 74 theme cellulose 1653:1661 arg1 degradation 1638:1648 the efficient degradation 1624:1648 the efficient degradation of cellulose in the presence of endoglucanase 1624:1694 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 1 75 theme important 243:251 arg1 route 253:257 an important route 240:257 an important route in green chemistry 240:276 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 1 75 theme important 243:251 arg1 degradation 207:217 their enzymatic degradation 191:217 their enzymatic degradation to soluble sugars 191:235 Structural polysaccharides like cellulose and chitin are abundant and their enzymatic degradation to soluble sugars is an important route in green chemistry. 27780868 0 76 theme Cellulose 60:68 arg1 Chain 70:74 Cellulose Chain 60:74 Cellulose Chain 60:74 Inter-domain Synergism Is Required for Efficient Feeding of Cellulose Chain into Active Site of Cellobiohydrolase Cel7A. 27780868 6 77 theme role 873:876 arg1 elucidation 854:864 the elucidation 850:864 the elucidation of the role of CBM and active site aromatics 850:909 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 27780868 12 78 theme efficient 1628:1636 arg1 degradation 1638:1648 the efficient degradation 1624:1648 the efficient degradation of cellulose in the presence of endoglucanase 1624:1694 The inter-domain synergism was pre-requisite for the efficient degradation of cellulose in the presence of endoglucanase. 27780868 6 79 theme multistep 942:950 arg1 mechanism 952:960 a complex multistep mechanism 932:960 a complex multistep mechanism of processive GHs 932:978 Although addressed in numerous studies the elucidation of the role of CBM and active site aromatics has been obscured by a complex multistep mechanism of processive GHs. 26075387 9 0 theme healthy 1394:1400 arg1 controls 1402:1409 healthy controls 1394:1409 healthy controls 1394:1409 Serum SA level was significantly higher in patients with LCMM than that in healthy controls. 26075387 4 1 theme core 519:522 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 2 2 theme IgG 281:283 arg1 myeloma 285:291 42 IgG myeloma 278:291 42 IgG myeloma 278:291 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. 26075387 6 3 theme N-glycan 792:799 arg1 structures 801:810 two N-glycan structures 788:810 two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) 788:840 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 6 4 dep structures 801:810 arg1 Peak7 835:839 Peak7 835:839 Peak7 835:839 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 6 4 dep structures 801:810 arg1 Peak3 821:825 Peak3 821:825 Peak3 821:825 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 5 5 theme sialic 722:727 arg1 SA 735:736 SA 735:736 SA 735:736 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 5 5 theme sialic 722:727 arg1 acid 729:732 serum sialic acid 716:732 serum sialic acid (SA) level 716:743 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 1 6 theme diagnostic 148:157 arg1 power 159:163 the diagnostic and differential diagnostic power 116:163 the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM) 116:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 7 7 dep sensitivity 971:981 arg1 The 967:969 The 967:969 The 967:969 The sensitivity and specificity of Peak3 were 88.1% and 92.9%, while Peak7 were 92.9% and 97.6%, respectively. 26075387 8 8 used used 1218:1221 arg2 Peak7 1203:1207 Peak7 1203:1207 Peak7 1203:1207 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 10 9 theme increased 1469:1477 arg1 sialylation 1479:1489 increased sialylation 1469:1489 increased sialylation 1469:1489 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 1 10 theme N-glycans 174:182 arg1 power 159:163 the diagnostic and differential diagnostic power 116:163 the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM) 116:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 11 11 from target 1741:1746 arg1 LCMM 1751:1754 LCMM 1751:1754 LCMM 1751:1754 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 1 12 theme light 188:192 arg1 chain 194:198 light chain multiple myeloma (LCMM) 188:222 light chain multiple myeloma (LCMM) 188:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 8 13 theme Peak3 1095:1099 arg1 abundance 1082:1090 The abundance 1078:1090 The abundance of Peak3 1078:1099 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 11 14 theme serum 1580:1584 arg1 N-glycan 1586:1593 serum N-glycan 1580:1593 serum N-glycan 1580:1593 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 11 15 theme potential 1719:1727 arg1 target 1741:1746 a new potential therapeutic target 1713:1746 a new potential therapeutic target in LCMM 1713:1754 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 1 16 theme study 94:98 arg1 aim 82:84 The aim 78:84 The aim of this study 78:98 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 3 17 theme quantitive 474:483 arg1 abundance 485:493 the quantitive abundance 470:493 the quantitive abundance of serum N-glycans 470:512 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 3 18 theme sequencer-assisted 368:385 arg1 DSA-FACE 435:442 DSA-FACE 435:442 DSA-FACE 435:442 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 3 18 theme sequencer-assisted 368:385 arg1 electrophoresis 418:432 sequencer-assisted fluorophore-assisted capillary electrophoresis 368:432 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) 364:443 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 0 19 theme Light 48:52 arg1 Myeloma 69:75 Light Chain Multiple Myeloma 48:75 Light Chain Multiple Myeloma 48:75 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 8 20 theme AUC 1267:1269 arg1 0.922 1271:1275 AUC 0.922 1267:1275 AUC 0.922 1267:1275 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 4 21 theme healthy 627:633 arg1 n=20 645:648 n=20 645:648 n=20 645:648 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 4 21 theme healthy 627:633 arg1 controls 635:642 healthy controls 627:642 healthy controls (n=20) 627:649 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 0 22 theme Multiple 60:67 arg1 Myeloma 69:75 Light Chain Multiple Myeloma 48:75 Light Chain Multiple Myeloma 48:75 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 6 23 theme diagnostic 861:870 arg1 efficacy 872:879 the optimum diagnostic efficacy 849:879 the optimum diagnostic efficacy 849:879 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 6 24 dep curve 905:909 arg1 0.939 917:921 0.939 917:921 0.939 917:921 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 6 24 dep curve 905:909 arg1 0.940 927:931 0.940 927:931 0.940 927:931 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 8 25 from myeloma 1254:1260 arg1 LCMM 1240:1243 LCMM 1240:1243 LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9% 1240:1316 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 4 26 gly sialylated 551:560 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 3 27 theme fluorophore-assisted 387:406 arg1 DSA-FACE 435:442 DSA-FACE 435:442 DSA-FACE 435:442 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 3 27 theme fluorophore-assisted 387:406 arg1 electrophoresis 418:432 sequencer-assisted fluorophore-assisted capillary electrophoresis 368:432 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) 364:443 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 4 28 theme lectin 693:698 arg1 blot 700:703 lectin blot 693:703 lectin blot 693:703 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 8 29 from myeloma 1135:1141 arg1 % 1196:1196 specificity 76.2% 1180:1196 specificity 76.2% 1180:1196 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 29 from myeloma 1135:1141 arg1 % 1174:1174 sensitivity 100% 1159:1174 sensitivity 100% 1159:1174 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 29 from myeloma 1135:1141 arg1 LCMM 1121:1124 LCMM 1121:1124 LCMM from IgG myeloma with AUC 0.899 1121:1156 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 9 30 theme Serum 1319:1323 arg1 level 1328:1332 Serum SA level 1319:1332 Serum SA level 1319:1332 Serum SA level was significantly higher in patients with LCMM than that in healthy controls. 26075387 0 31 theme Serum 0:4 arg1 N-Glycans 6:14 Serum N-Glycans 0:14 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 0:76 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 4 32 gly fucosylated 524:534 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 10 33 theme core 1436:1439 arg1 fucosylation 1441:1452 core fucosylation 1436:1452 core fucosylation 1436:1452 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 2 34 theme healthy 317:323 arg1 controls 325:332 42 healthy controls 314:332 42 healthy controls 314:332 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. 26075387 7 35 theme Peak3 1002:1006 arg1 specificity 987:997 specificity 987:997 specificity 987:997 The sensitivity and specificity of Peak3 were 88.1% and 92.9%, while Peak7 were 92.9% and 97.6%, respectively. 26075387 7 35 theme Peak3 1002:1006 arg1 sensitivity 971:981 sensitivity 971:981 sensitivity 971:981 The sensitivity and specificity of Peak3 were 88.1% and 92.9%, while Peak7 were 92.9% and 97.6%, respectively. 26075387 6 36 theme ROC 901:903 arg1 AUC 912:914 AUC 912:914 AUC 912:914 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 6 36 theme ROC 901:903 arg1 curve 905:909 the ROC curve 897:909 the ROC curve (AUC) 0.939 and 0.940 897:931 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 8 37 with LCMM 1240:1243 arg1 % 1294:1294 sensitivity 92.9% 1278:1294 sensitivity 92.9% 1278:1294 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 37 with LCMM 1240:1243 arg1 0.922 1271:1275 AUC 0.922 1267:1275 AUC 0.922 1267:1275 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 37 with LCMM 1240:1243 arg1 % 1316:1316 specificity 82.9% 1300:1316 specificity 82.9% 1300:1316 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 3 38 theme capillary 408:416 arg1 DSA-FACE 435:442 DSA-FACE 435:442 DSA-FACE 435:442 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 3 38 theme capillary 408:416 arg1 electrophoresis 418:432 sequencer-assisted fluorophore-assisted capillary electrophoresis 368:432 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) 364:443 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 10 39 theme serum 1505:1509 arg1 glycoproteins 1511:1523 serum glycoproteins 1505:1523 serum glycoproteins 1505:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 3 40 dep DNA 364:366 arg1 DSA-FACE 435:442 DSA-FACE 435:442 DSA-FACE 435:442 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 3 40 dep DNA 364:366 arg1 electrophoresis 418:432 sequencer-assisted fluorophore-assisted capillary electrophoresis 368:432 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) 364:443 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 4 41 theme bisecting 537:545 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 8 42 with LCMM 1121:1124 arg1 AUC 1148:1150 AUC 0.899 1148:1156 AUC 0.899 1148:1156 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 11 43 dep potential 1719:1727 arg1 therapeutic 1729:1739 therapeutic 1729:1739 therapeutic 1729:1739 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 6 44 dep found 782:786 arg1 showed 842:847 showed 842:847 found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control 782:964 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 10 45 theme sialylation 1479:1489 arg1 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 9 46 with patients 1362:1369 arg1 LCMM 1376:1379 LCMM 1376:1379 LCMM 1376:1379 Serum SA level was significantly higher in patients with LCMM than that in healthy controls. 26075387 2 47 theme cases 240:244 arg1 total 227:231 A total 225:231 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls 225:332 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. 26075387 4 48 theme fucosylated 524:534 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 10 49 theme glycoproteins 1511:1523 arg1 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 1 50 theme differential 135:146 arg1 power 159:163 the diagnostic and differential diagnostic power 116:163 the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM) 116:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 3 51 theme N-glycans 504:512 arg1 abundance 485:493 the quantitive abundance 470:493 the quantitive abundance of serum N-glycans 470:512 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 5 52 theme enzymatic 761:769 arg1 method 771:776 enzymatic method 761:776 enzymatic method 761:776 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 4 53 theme patients 607:614 arg1 serum 593:597 serum 593:597 serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort 593:688 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 2 54 theme subjects 249:256 arg1 cases 240:244 167 cases 236:244 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls 236:332 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. 26075387 1 55 theme serum 168:172 arg1 N-glycans 174:182 serum N-glycans 168:182 serum N-glycans 168:182 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 6 56 dep Peak3 821:825 arg1 NA2FB 828:832 NA2FB 828:832 NA2FB 828:832 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 10 57 theme bisecting 1455:1463 arg1 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 1 58 dep chain 194:198 arg1 myeloma 209:215 multiple myeloma 200:215 light chain multiple myeloma (LCMM) 188:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 1 58 dep chain 194:198 arg1 LCMM 218:221 LCMM 218:221 LCMM 218:221 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 11 59 theme new 1715:1717 arg1 target 1741:1746 a new potential therapeutic target 1713:1746 a new potential therapeutic target in LCMM 1713:1754 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 8 60 theme IgA 1250:1252 arg1 myeloma 1254:1260 IgA myeloma 1250:1260 IgA myeloma 1250:1260 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 4 61 theme controls 635:642 arg1 serum 593:597 serum 593:597 serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort 593:688 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 0 62 theme Chain 54:58 arg1 Myeloma 69:75 Light Chain Multiple Myeloma 48:75 Light Chain Multiple Myeloma 48:75 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 10 63 located observed 1530:1537 arg1 patients 1542:1549 patients 1542:1549 patients with LCMM 1542:1559 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 10 63 located observed 1530:1537 arg2 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 11 64 theme non-invasive 1632:1643 arg1 biomarker 1645:1653 a simple, reliable and non-invasive biomarker 1609:1653 a simple, reliable and non-invasive biomarker for LCMM diagnosis 1609:1672 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 1 65 theme diagnostic 120:129 arg1 power 159:163 the diagnostic and differential diagnostic power 116:163 the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM) 116:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 10 66 with patients 1542:1549 arg1 LCMM 1556:1559 LCMM 1556:1559 LCMM 1556:1559 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 8 67 theme sensitivity 1278:1288 arg1 % 1294:1294 sensitivity 92.9% 1278:1294 sensitivity 92.9% 1278:1294 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 68 theme IgG 1131:1133 arg1 myeloma 1135:1141 IgG myeloma 1131:1141 IgG myeloma 1131:1141 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 3 69 theme serum 498:502 arg1 N-glycans 504:512 serum N-glycans 498:512 serum N-glycans 498:512 DNA sequencer-assisted fluorophore-assisted capillary electrophoresis (DSA-FACE) was applied to determine the quantitive abundance of serum N-glycans. 26075387 6 70 theme optimum 853:859 arg1 efficacy 872:879 the optimum diagnostic efficacy 849:879 the optimum diagnostic efficacy 849:879 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 4 71 theme same 678:681 arg1 cohort 683:688 the same cohort 674:688 the same cohort 674:688 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26075387 10 72 theme fucosylation 1441:1452 arg1 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 8 73 theme specificity 1300:1310 arg1 % 1316:1316 specificity 82.9% 1300:1316 specificity 82.9% 1300:1316 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 11 74 theme abnormal 1678:1685 arg1 glycosylation 1687:1699 abnormal glycosylation 1678:1699 abnormal glycosylation 1678:1699 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 5 75 theme serum 716:720 arg1 SA 735:736 SA 735:736 SA 735:736 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 5 75 theme serum 716:720 arg1 acid 729:732 serum sialic acid 716:732 serum sialic acid (SA) level 716:743 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 9 76 theme SA 1325:1326 arg1 level 1328:1332 Serum SA level 1319:1332 Serum SA level 1319:1332 Serum SA level was significantly higher in patients with LCMM than that in healthy controls. 26075387 8 77 theme sensitivity 1159:1169 arg1 % 1174:1174 sensitivity 100% 1159:1174 sensitivity 100% 1159:1174 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 77 theme sensitivity 1159:1169 arg1 LCMM 1121:1124 LCMM 1121:1124 LCMM from IgG myeloma with AUC 0.899 1121:1156 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 2 78 theme IgA 297:299 arg1 myeloma 301:307 41 IgA myeloma 294:307 41 IgA myeloma 294:307 A total of 167 cases of subjects, including 42 LCMM, 42 IgG myeloma, 41 IgA myeloma, and 42 healthy controls were recruited in this study. 26075387 5 79 theme acid 729:732 arg1 level 739:743 serum sialic acid (SA) level 716:743 serum sialic acid (SA) level 716:743 Moreover, serum sialic acid (SA) level was measured by enzymatic method. 26075387 10 80 theme decreased 1426:1434 arg1 characters 1491:1500 the decreased core fucosylation, bisecting and increased sialylation characters 1422:1500 the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins 1422:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 0 81 theme Diagnostic 23:32 arg1 Biomarker 34:42 A New Diagnostic Biomarker 17:42 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 0:76 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 8 82 theme specificity 1180:1190 arg1 % 1196:1196 specificity 76.2% 1180:1196 specificity 76.2% 1180:1196 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 8 82 theme specificity 1180:1190 arg1 LCMM 1121:1124 LCMM 1121:1124 LCMM from IgG myeloma with AUC 0.899 1121:1156 The abundance of Peak3 could differentiate LCMM from IgG myeloma with AUC 0.899, sensitivity 100% and specificity 76.2%, and Peak7 could be used to differentiate LCMM from IgA myeloma with AUC 0.922, sensitivity 92.9% and specificity 82.9%. 26075387 11 83 theme reliable 1619:1626 arg1 biomarker 1645:1653 a simple, reliable and non-invasive biomarker 1609:1653 a simple, reliable and non-invasive biomarker for LCMM diagnosis 1609:1672 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 10 84 gly glycoproteins 1511:1523 arg1 glycoproteins 1511:1523 serum glycoproteins 1505:1523 serum glycoproteins 1505:1523 Moreover, the decreased core fucosylation, bisecting and increased sialylation characters of serum glycoproteins were observed in patients with LCMM. 26075387 11 85 theme simple 1611:1616 arg1 biomarker 1645:1653 a simple, reliable and non-invasive biomarker 1609:1653 a simple, reliable and non-invasive biomarker for LCMM diagnosis 1609:1672 We concluded that serum N-glycan could provide a simple, reliable and non-invasive biomarker for LCMM diagnosis and abnormal glycosylation might imply a new potential therapeutic target in LCMM. 26075387 1 86 theme multiple 200:207 arg1 myeloma 209:215 multiple myeloma 200:215 light chain multiple myeloma (LCMM) 188:222 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 1 86 theme multiple 200:207 arg1 LCMM 218:221 LCMM 218:221 LCMM 218:221 The aim of this study was to evaluate the diagnostic and differential diagnostic power of serum N-glycans for light chain multiple myeloma (LCMM). 26075387 0 87 dep N-Glycans 6:14 arg1 Biomarker 34:42 A New Diagnostic Biomarker 17:42 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 0:76 Serum N-Glycans: A New Diagnostic Biomarker for Light Chain Multiple Myeloma. 26075387 6 88 theme healthy 950:956 arg1 control 958:964 healthy control 950:964 healthy control 950:964 We found two N-glycan structures (NG1A2F, Peak3; NA2FB, Peak7) showed the optimum diagnostic efficacy with area under the ROC curve (AUC) 0.939 and 0.940 between LCMM and healthy control. 26075387 4 89 theme sialylated 551:560 arg1 modifications 562:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications 515:574 The core fucosylated, bisecting and sialylated modifications were analyzed in serum of LCMM patients (n=20) and healthy controls (n=20) randomly selected from the same cohort by lectin blot. 26162743 2 0 theme regioselective 400:413 arg1 glycosylation 415:427 the regioselective glycosylation 396:427 the regioselective glycosylation of a GlcNAc acceptor 396:448 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 1 theme free 530:533 arg1 hydroxyl 542:549 the remaining free GlcNAc hydroxyl 516:549 the remaining free GlcNAc hydroxyl 516:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 2 theme groups 730:735 arg1 nature 705:710 the orthogonal nature 690:710 the orthogonal nature of the protecting groups employed 690:744 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 0 3 theme crystalline 47:57 arg1 intermediate 59:70 a late-stage crystalline intermediate 34:70 a late-stage crystalline intermediate 34:70 Total synthesis of Lewis(X) using a late-stage crystalline intermediate. 26162743 2 4 theme remaining 520:528 arg1 hydroxyl 542:549 the remaining free GlcNAc hydroxyl 516:549 the remaining free GlcNAc hydroxyl 516:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 5 theme acceptor 441:448 arg1 glycosylation 415:427 the regioselective glycosylation 396:427 the regioselective glycosylation of a GlcNAc acceptor 396:448 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 6 theme protecting 719:728 arg1 groups 730:735 the protecting groups 715:735 the protecting groups employed 715:744 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 2 7 theme GlcNAc 434:439 arg1 acceptor 441:448 a GlcNAc acceptor 432:448 a GlcNAc acceptor 432:448 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 8 theme key 554:556 arg1 strategy 373:380 a strategy 371:380 a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl 371:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 8 theme key 554:556 arg1 steps 558:562 key steps 554:562 key steps 554:562 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 9 theme linear 321:326 arg1 sequence 328:335 longest linear sequence 313:335 longest linear sequence 313:335 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 9 theme linear 321:326 arg1 steps 306:310 seven steps 300:310 seven steps 300:310 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 1 10 theme crystalline 128:138 arg1 trisaccharide 159:171 a crystalline protected Lewis(X) trisaccharide 126:171 a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 126:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 1 11 theme protected 140:148 arg1 trisaccharide 159:171 a crystalline protected Lewis(X) trisaccharide 126:171 a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 126:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 3 12 theme other 630:634 arg1 members 636:642 other members 630:642 other members of the Type-2 Lewis family of antigens 630:681 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 2 13 theme hydroxyl 542:549 arg1 fucosylation 500:511 fucosylation 500:511 fucosylation of the remaining free GlcNAc hydroxyl 500:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 14 theme Lewis 658:662 arg1 family 664:669 the Type-2 Lewis family 647:669 the Type-2 Lewis family of antigens 647:681 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 2 15 gly glycosylation 415:427 arg1 acceptor 441:448 a GlcNAc acceptor 432:448 a GlcNAc acceptor 432:448 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 1 16 theme Lewis 150:154 arg1 trisaccharide 159:171 a crystalline protected Lewis(X) trisaccharide 126:171 a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 126:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 0 17 theme Total 0:4 arg1 synthesis 6:14 Total synthesis 0:14 Total synthesis of Lewis(X) 0:26 Total synthesis of Lewis(X) using a late-stage crystalline intermediate. 26162743 1 18 theme global 214:219 arg1 deprotection 221:232 global deprotection 214:232 global deprotection 214:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 2 19 theme GlcNAc 535:540 arg1 hydroxyl 542:549 the remaining free GlcNAc hydroxyl 516:549 the remaining free GlcNAc hydroxyl 516:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 20 theme longest 313:319 arg1 sequence 328:335 longest linear sequence 313:335 longest linear sequence 313:335 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 20 theme longest 313:319 arg1 steps 306:310 seven steps 300:310 seven steps 300:310 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 0 21 theme Lewis 19:23 arg1 synthesis 6:14 Total synthesis 0:14 Total synthesis of Lewis(X) 0:26 Total synthesis of Lewis(X) using a late-stage crystalline intermediate. 26162743 3 22 theme family 664:669 arg1 members 636:642 other members 630:642 other members of the Type-2 Lewis family of antigens 630:681 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 2 23 dep synthesis 289:297 arg1 sequence 328:335 longest linear sequence 313:335 longest linear sequence 313:335 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 23 dep synthesis 289:297 arg1 steps 306:310 seven steps 300:310 seven steps 300:310 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 24 theme 38 347:348 arg1 % 349:349 % 349:349 % 349:349 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 25 theme % 349:349 arg1 yield 359:363 a 38% overall yield 345:363 a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps 345:562 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 26 theme thioglycoside 467:479 arg1 donor 481:485 a galactose thioglycoside donor 455:485 a galactose thioglycoside donor 455:485 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 27 theme convergent 278:287 arg1 synthesis 289:297 a highly convergent synthesis 269:297 a highly convergent synthesis (seven steps, longest linear sequence) 269:336 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 2 28 theme galactose 457:465 arg1 donor 481:485 a galactose thioglycoside donor 455:485 a galactose thioglycoside donor 455:485 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 29 theme core 569:572 arg1 trisaccharide 574:586 The core trisaccharide 565:586 The core trisaccharide 565:586 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 3 30 contain has 593:595 arg2 potential 601:609 the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed 597:744 the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed 597:744 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 3 30 contain has 593:595 arg1 trisaccharide 574:586 The core trisaccharide 565:586 The core trisaccharide 565:586 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 1 31 theme trisaccharide 159:171 arg1 synthesis 113:121 a highly efficient synthesis 94:121 a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 94:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 2 32 theme overall 351:357 arg1 yield 359:363 a 38% overall yield 345:363 a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps 345:562 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 33 theme Type-2 651:656 arg1 family 664:669 the Type-2 Lewis family 647:669 the Type-2 Lewis family of antigens 647:681 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 2 34 gly fucosylation 500:511 arg1 hydroxyl 542:549 the remaining free GlcNAc hydroxyl 516:549 the remaining free GlcNAc hydroxyl 516:549 The trisaccharide was prepared in a highly convergent synthesis (seven steps, longest linear sequence) and in a 38% overall yield using a strategy that involved the regioselective glycosylation of a GlcNAc acceptor with a galactose thioglycoside donor, followed by fucosylation of the remaining free GlcNAc hydroxyl as key steps. 26162743 3 35 theme antigens 674:681 arg1 family 664:669 the Type-2 Lewis family 647:669 the Type-2 Lewis family of antigens 647:681 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26162743 0 36 theme late-stage 36:45 arg1 intermediate 59:70 a late-stage crystalline intermediate 34:70 a late-stage crystalline intermediate 34:70 Total synthesis of Lewis(X) using a late-stage crystalline intermediate. 26162743 1 37 theme X 156:156 arg1 trisaccharide 159:171 a crystalline protected Lewis(X) trisaccharide 126:171 a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 126:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 1 38 theme efficient 103:111 arg1 synthesis 113:121 a highly efficient synthesis 94:121 a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection 94:232 Herein, we report on a highly efficient synthesis of a crystalline protected Lewis(X) trisaccharide that was converted to Lewis(X) following global deprotection. 26162743 3 39 theme orthogonal 694:703 arg1 nature 705:710 the orthogonal nature 690:710 the orthogonal nature of the protecting groups employed 690:744 The core trisaccharide also has the potential to be converted to other members of the Type-2 Lewis family of antigens due to the orthogonal nature of the protecting groups employed. 26626878 0 0 theme biofilm 80:86 arg1 matrix 88:93 the Listeria monocytogenes biofilm matrix 53:93 the Listeria monocytogenes biofilm matrix 53:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 2 1 from matrix 433:438 arg1 embedded 404:411 embedded 404:411 embedded 404:411 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 1 2 from biofilm 165:171 arg1 matrix 251:256 the biofilm extracellular matrix 225:256 the biofilm extracellular matrix 225:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 3 theme extracellular 237:249 arg1 matrix 251:256 the biofilm extracellular matrix 225:256 the biofilm extracellular matrix 225:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 0 4 theme monocytogenes 66:78 arg1 matrix 88:93 the Listeria monocytogenes biofilm matrix 53:93 the Listeria monocytogenes biofilm matrix 53:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 1 5 theme Listeria 142:149 arg1 nature 194:199 the nature 190:199 the nature of the carbohydrates in the biofilm extracellular matrix 190:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 5 theme Listeria 142:149 arg1 monocytogenes 151:163 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 5 theme Listeria 142:149 arg1 biofilm 165:171 biofilm 165:171 biofilm 165:171 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 5 6 contain have 863:866 arg1 acids 828:832 the extracellular teichoic acids 801:832 the extracellular teichoic acids in L. monocytogenes biofilms 801:861 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 5 6 contain have 863:866 arg2 origin 877:882 the same origin 868:882 the same origin 868:882 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 5 7 theme teichoic 897:904 arg1 acid 906:909 cell wall teichoic acid 887:909 cell wall teichoic acid 887:909 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 3 8 theme cell 586:589 arg1 acids 605:609 cell wall teichoic acids 586:609 cell wall teichoic acids 586:609 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 9 theme wall 591:594 arg1 acids 605:609 cell wall teichoic acids 586:609 cell wall teichoic acids 586:609 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 10 theme identical 573:581 arg1 acids 553:557 teichoic acids 544:557 teichoic acids 544:557 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 10 theme identical 573:581 arg1 carbohydrates 453:465 The soluble carbohydrates 441:465 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant 441:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 1 11 theme culture 262:268 arg1 supernatant 270:280 culture supernatant 262:280 culture supernatant 262:280 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 4 12 theme glucosamine 701:711 arg1 glycosylation 713:725 N-acetyl glucosamine glycosylation 692:725 N-acetyl glucosamine glycosylation 692:725 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 0 13 from present 42:48 arg1 matrix 88:93 the Listeria monocytogenes biofilm matrix 53:93 the Listeria monocytogenes biofilm matrix 53:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 4 14 theme BFA2 638:641 arg1 strain 643:648 the DSS 1130 BFA2 strain 625:648 the DSS 1130 BFA2 strain 625:648 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 4 15 theme 1/2a 665:668 arg1 acid 679:682 a serotype 1/2a teichoic acid 654:682 a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene 654:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 5 16 theme extracellular 805:817 arg1 acids 828:832 the extracellular teichoic acids 801:832 the extracellular teichoic acids in L. monocytogenes biofilms 801:861 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 5 17 theme monocytogenes 840:852 arg1 biofilms 854:861 L. monocytogenes biofilms 837:861 L. monocytogenes biofilms 837:861 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 3 18 from matrix 494:499 arg1 acids 553:557 teichoic acids 544:557 teichoic acids 544:557 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 18 from matrix 494:499 arg1 carbohydrates 453:465 The soluble carbohydrates 441:465 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant 441:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 2 19 theme complex 387:393 arg1 biofilms 395:402 complex biofilms 387:402 complex biofilms embedded in an extracellular matrix 387:438 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 5 20 theme teichoic 819:826 arg1 acids 828:832 the extracellular teichoic acids 801:832 the extracellular teichoic acids in L. monocytogenes biofilms 801:861 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 5 21 from acids 828:832 arg1 biofilms 854:861 L. monocytogenes biofilms 837:861 L. monocytogenes biofilms 837:861 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 4 22 theme serotype 656:663 arg1 acid 679:682 a serotype 1/2a teichoic acid 654:682 a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene 654:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 2 23 theme Listeria 317:324 arg1 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains 317:367 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 0 24 theme Teichoic 0:7 arg1 polysaccharide 27:40 the major polysaccharide 17:40 the major polysaccharide present in the Listeria monocytogenes biofilm matrix 17:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 24 theme Teichoic 0:7 arg1 acid 9:12 Teichoic acid 0:12 Teichoic acid 0:12 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 25 attach present 42:48 arg2 polysaccharide 27:40 the major polysaccharide 17:40 the major polysaccharide present in the Listeria monocytogenes biofilm matrix 17:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 25 attach present 42:48 arg2 acid 9:12 Teichoic acid 0:12 Teichoic acid 0:12 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 25 attach present 42:48 arg1 matrix 88:93 the Listeria monocytogenes biofilm matrix 53:93 the Listeria monocytogenes biofilm matrix 53:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 3 26 theme teichoic 596:603 arg1 acids 605:609 cell wall teichoic acids 586:609 cell wall teichoic acids 586:609 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 4 27 theme teichoic 670:677 arg1 acid 679:682 a serotype 1/2a teichoic acid 654:682 a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene 654:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 5 28 theme same 872:875 arg1 origin 877:882 the same origin 868:882 the same origin 868:882 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 3 29 from supernatant 513:523 arg1 acids 553:557 teichoic acids 544:557 teichoic acids 544:557 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 29 from supernatant 513:523 arg1 carbohydrates 453:465 The soluble carbohydrates 441:465 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant 441:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 2 30 theme extracellular 419:431 arg1 matrix 433:438 an extracellular matrix 416:438 an extracellular matrix 416:438 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 3 31 theme biofilm 472:478 arg1 matrix 494:499 biofilm extracellular matrix 472:499 biofilm extracellular matrix 472:499 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 4 32 contain had 650:652 arg2 acid 679:682 a serotype 1/2a teichoic acid 654:682 a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene 654:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 4 32 contain had 650:652 arg1 strain 643:648 the DSS 1130 BFA2 strain 625:648 the DSS 1130 BFA2 strain 625:648 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 2 33 theme 4b 358:359 arg1 strains 361:367 Listeria monocytogenes serotype 1/2a and 4b strains 317:367 strains 361:367 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 3 34 theme extracellular 480:492 arg1 matrix 494:499 biofilm extracellular matrix 472:499 biofilm extracellular matrix 472:499 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 1 35 theme cell 292:295 arg1 carbohydrates 302:314 cell wall carbohydrates 292:314 cell wall carbohydrates 292:314 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 5 36 theme cell 887:890 arg1 acid 906:909 cell wall teichoic acid 887:909 cell wall teichoic acid 887:909 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 0 37 theme major 21:25 arg1 polysaccharide 27:40 the major polysaccharide 17:40 the major polysaccharide present in the Listeria monocytogenes biofilm matrix 17:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 37 theme major 21:25 arg1 acid 9:12 Teichoic acid 0:12 Teichoic acid 0:12 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 1 38 theme wall 297:300 arg1 carbohydrates 302:314 cell wall carbohydrates 292:314 cell wall carbohydrates 292:314 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 39 from matrix 251:256 arg1 nature 194:199 the nature 190:199 the nature of the carbohydrates in the biofilm extracellular matrix 190:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 39 from matrix 251:256 arg1 monocytogenes 151:163 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 39 from matrix 251:256 arg1 biofilm 165:171 biofilm 165:171 biofilm 165:171 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 2 40 theme embedded 404:411 arg1 biofilms 395:402 complex biofilms 387:402 complex biofilms embedded in an extracellular matrix 387:438 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 5 41 theme wall 892:895 arg1 acid 906:909 cell wall teichoic acid 887:909 cell wall teichoic acid 887:909 Consequently, we hypothesized that the extracellular teichoic acids in L. monocytogenes biofilms have the same origin as cell wall teichoic acid. 26626878 1 42 dep monocytogenes 151:163 arg1 nature 194:199 the nature 190:199 the nature of the carbohydrates in the biofilm extracellular matrix 190:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 42 dep monocytogenes 151:163 arg1 monocytogenes 151:163 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 42 dep monocytogenes 151:163 arg1 biofilm 165:171 biofilm 165:171 biofilm 165:171 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 3 43 theme culture 505:511 arg1 supernatant 513:523 culture supernatant 505:523 culture supernatant 505:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 0 44 theme present 42:48 arg1 polysaccharide 27:40 the major polysaccharide 17:40 the major polysaccharide present in the Listeria monocytogenes biofilm matrix 17:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 44 theme present 42:48 arg1 acid 9:12 Teichoic acid 0:12 Teichoic acid 0:12 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 4 45 from mutation 736:743 arg1 gene 760:763 the lmo2550 gene 748:763 the lmo2550 gene 748:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 1 46 from carbohydrates 208:220 arg1 matrix 251:256 the biofilm extracellular matrix 225:256 the biofilm extracellular matrix 225:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 47 from nature 194:199 arg1 matrix 251:256 the biofilm extracellular matrix 225:256 the biofilm extracellular matrix 225:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 48 theme study 112:116 arg1 aim 100:102 The aim 96:102 The aim of this study 96:116 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 49 theme carbohydrates 208:220 arg1 nature 194:199 the nature 190:199 the nature of the carbohydrates in the biofilm extracellular matrix 190:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 49 theme carbohydrates 208:220 arg1 monocytogenes 151:163 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix 138:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 1 49 theme carbohydrates 208:220 arg1 biofilm 165:171 biofilm 165:171 biofilm 165:171 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 26626878 4 50 theme lmo2550 752:758 arg1 gene 760:763 the lmo2550 gene 748:763 the lmo2550 gene 748:763 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 2 51 theme monocytogenes 326:338 arg1 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains 317:367 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 2 52 theme serotype 340:347 arg1 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains 317:367 1/2a 349:352 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 0 53 theme Listeria 57:64 arg1 monocytogenes 66:78 Listeria monocytogenes 57:78 the Listeria monocytogenes biofilm matrix 53:93 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 0 54 from matrix 88:93 arg1 present 42:48 present 42:48 present 42:48 Teichoic acid is the major polysaccharide present in the Listeria monocytogenes biofilm matrix. 26626878 2 55 from embedded 404:411 arg1 matrix 433:438 an extracellular matrix 416:438 an extracellular matrix 416:438 Listeria monocytogenes serotype 1/2a and 4b strains were able to form complex biofilms embedded in an extracellular matrix. 26626878 4 56 theme N-acetyl 692:699 arg1 glycosylation 713:725 N-acetyl glucosamine glycosylation 692:725 N-acetyl glucosamine glycosylation 692:725 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 3 57 theme soluble 445:451 arg1 acids 553:557 teichoic acids 544:557 teichoic acids 544:557 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 57 theme soluble 445:451 arg1 carbohydrates 453:465 The soluble carbohydrates 441:465 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant 441:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 4 58 theme DSS 629:631 arg1 strain 643:648 the DSS 1130 BFA2 strain 625:648 the DSS 1130 BFA2 strain 625:648 In addition, the DSS 1130 BFA2 strain had a serotype 1/2a teichoic acid lacking N-acetyl glucosamine glycosylation due to a mutation in the lmo2550 gene. 26626878 3 59 theme teichoic 544:551 arg1 acids 553:557 teichoic acids 544:557 teichoic acids 544:557 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 3 59 theme teichoic 544:551 arg1 carbohydrates 453:465 The soluble carbohydrates 441:465 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant 441:523 The soluble carbohydrates from biofilm extracellular matrix and culture supernatant were identified as teichoic acids, structurally identical to cell wall teichoic acids. 26626878 1 60 theme biofilm 229:235 arg1 matrix 251:256 the biofilm extracellular matrix 225:256 the biofilm extracellular matrix 225:256 The aim of this study was to characterize the Listeria monocytogenes biofilm and particularly the nature of the carbohydrates in the biofilm extracellular matrix and culture supernatant versus to cell wall carbohydrates. 28097217 3 0 theme glycoside 296:304 arg1 GHs 318:320 GHs 318:320 GHs 318:320 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 3 0 theme glycoside 296:304 arg1 hydrolases 306:315 glycoside hydrolases 296:315 glycoside hydrolases (GHs) 296:321 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 7 1 dep a 1023:1023 arg1 mimic 1025:1029 mimic 1025:1029 mimic 1025:1029 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 8 2 with crevice 1146:1152 arg1 length 1161:1166 a length 1159:1166 a length of more than 40 Å 1159:1184 In total, 15 glucose residues interact with the protein in an extended crevice with a length of more than 40 Å. 28097217 6 3 from data 677:680 arg1 binding 707:713 short oligosaccharide binding 685:713 short oligosaccharide binding to GHs 685:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 9 4 theme amino 1322:1326 arg1 acid 1328:1331 this nonstandard amino acid 1305:1331 this nonstandard amino acid 1305:1331 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 8 5 theme extended 1137:1144 arg1 crevice 1146:1152 an extended crevice 1134:1152 an extended crevice with a length of more than 40 Å 1134:1184 In total, 15 glucose residues interact with the protein in an extended crevice with a length of more than 40 Å. 28097217 7 6 theme AM 919:920 arg1 structure 888:896 the crystal structure 876:896 the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 876:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 10 7 theme minimal 1510:1516 arg1 determination 1528:1540 minimal ring-size determination 1510:1540 minimal ring-size determination of CA products 1510:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 7 8 theme crystal 880:886 arg1 structure 888:896 the crystal structure 876:896 the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 876:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 7 9 theme amylose 1056:1062 arg1 substrate 1064:1072 the natural polymeric amylose substrate 1034:1072 the natural polymeric amylose substrate 1034:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 5 10 theme polymeric 570:578 arg1 rings 586:590 polymeric sugar rings 570:590 polymeric sugar rings 570:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 3 11 theme great 329:333 arg1 importance 359:368 great biological and societal importance 329:368 great biological and societal importance 329:368 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 4 12 theme sugar 445:449 arg1 chains 451:456 α-1,4-linked sugar chains 432:456 α-1,4-linked sugar chains such as amylose 432:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 4 12 theme sugar 445:449 arg1 amylose 466:472 amylose 466:472 amylose 466:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 6 13 theme oligosaccharide 691:705 arg1 binding 707:713 short oligosaccharide binding 685:713 short oligosaccharide binding to GHs 685:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 5 14 theme sugar 580:584 arg1 rings 586:590 polymeric sugar rings 570:590 polymeric sugar rings 570:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 9 15 attach derived 1211:1217 arg2 succinimide 1198:1208 A modified succinimide 1187:1208 A modified succinimide 1187:1208 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 9 15 attach derived 1211:1217 arg1 aspartate 1224:1232 aspartate 1224:1232 aspartate 1224:1232 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 4 16 theme α-1,4-linked 432:443 arg1 chains 451:456 α-1,4-linked sugar chains 432:456 α-1,4-linked sugar chains such as amylose 432:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 4 16 theme α-1,4-linked 432:443 arg1 amylose 466:472 amylose 466:472 amylose 466:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 7 17 theme natural 1038:1044 arg1 substrate 1064:1072 the natural polymeric amylose substrate 1034:1072 the natural polymeric amylose substrate 1034:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 6 18 theme structural 726:735 arg1 evidence 737:744 no structural evidence 723:744 no structural evidence 723:744 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 10 19 theme ruler 1490:1494 arg1 mechanism 1496:1504 a molecular ruler mechanism 1478:1504 a molecular ruler mechanism for minimal ring-size determination of CA products 1478:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 7 20 theme polymeric 1046:1054 arg1 substrate 1064:1072 the natural polymeric amylose substrate 1034:1072 the natural polymeric amylose substrate 1034:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 1 21 theme major 77:81 arg1 Starch 65:70 Starch 65:70 Starch 65:70 Starch is a major carbon and energy source throughout all kingdoms of life. 28097217 1 21 theme major 77:81 arg1 carbon 83:88 a major carbon and energy source 75:106 carbon 83:88 Starch is a major carbon and energy source throughout all kingdoms of life. 28097217 3 22 theme societal 350:357 arg1 importance 359:368 great biological and societal importance 329:368 great biological and societal importance 329:368 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 4 23 theme chains 451:456 arg1 hydrolysis 418:427 the hydrolysis 414:427 the hydrolysis of α-1,4-linked sugar chains such as amylose 414:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 10 24 theme unique 1391:1396 arg1 insights 1398:1405 unique insights 1391:1405 unique insights into the interaction of GHs with their polymeric substrate 1391:1464 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 5 25 theme bound 543:547 arg1 chain 555:559 a bound sugar chain 541:559 a bound sugar chain yielding polymeric sugar rings 541:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 6 26 theme natural 833:839 arg1 polysaccharide 841:854 the natural polysaccharide 829:854 the natural polysaccharide 829:854 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 0 27 theme Amylose 0:6 arg1 recognition 8:18 Amylose recognition 0:18 Amylose recognition 0:18 Amylose recognition and ring-size determination of amylomaltase. 28097217 10 28 theme products 1548:1555 arg1 determination 1528:1540 minimal ring-size determination 1510:1540 minimal ring-size determination of CA products 1510:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 9 29 theme modified 1189:1196 arg1 succinimide 1198:1208 A modified succinimide 1187:1208 A modified succinimide 1187:1208 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 5 30 theme sugar 549:553 arg1 chain 555:559 a bound sugar chain 541:559 a bound sugar chain yielding polymeric sugar rings 541:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 8 31 theme glucose 1088:1094 arg1 residues 1096:1103 15 glucose residues 1085:1103 15 glucose residues 1085:1103 In total, 15 glucose residues interact with the protein in an extended crevice with a length of more than 40 Å. 28097217 3 32 theme biological 335:344 arg1 importance 359:368 great biological and societal importance 329:368 great biological and societal importance 329:368 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 7 33 theme Thermus 901:907 arg1 AM 919:920 Thermus aquaticus AM 901:920 Thermus aquaticus AM 901:920 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 5 34 theme glucose 642:648 arg1 units 650:654 20 to 100 glucose units 632:654 20 to 100 glucose units 632:654 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 5 35 theme chain 555:559 arg1 cycloamyloses 597:609 the cycloamyloses 593:609 the cycloamyloses (CAs) 593:615 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 5 35 theme chain 555:559 arg1 transglycosylation 519:536 an intramolecular transglycosylation 501:536 an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings 501:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 1 36 theme energy 94:99 arg1 source 101:106 a major carbon and energy source 75:106 source 101:106 Starch is a major carbon and energy source throughout all kingdoms of life. 28097217 7 37 theme largest 963:969 arg1 carbohydrates 971:983 the largest carbohydrates 959:983 the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 959:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 10 38 theme ring-size 1518:1526 arg1 determination 1528:1540 minimal ring-size determination 1510:1540 minimal ring-size determination of CA products 1510:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 6 39 from wealth 667:672 arg1 binding 707:713 short oligosaccharide binding 685:713 short oligosaccharide binding to GHs 685:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 5 40 gly transglycosylation 519:536 arg1 chain 555:559 a bound sugar chain 541:559 a bound sugar chain yielding polymeric sugar rings 541:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 2 41 theme carbohydrate 160:171 arg1 polymers 173:180 two carbohydrate polymers 156:180 two carbohydrate polymers 156:180 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 2 41 theme carbohydrate 160:171 arg1 amylopectin 192:202 branched amylopectin 183:202 branched amylopectin 183:202 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 2 41 theme carbohydrate 160:171 arg1 amylose 215:221 linear amylose 208:221 linear amylose 208:221 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 8 42 theme Å 1184:1184 arg1 length 1161:1166 a length 1159:1166 a length of more than 40 Å 1159:1184 In total, 15 glucose residues interact with the protein in an extended crevice with a length of more than 40 Å. 28097217 2 43 from soluble 244:250 arg1 water 255:259 water 255:259 water 255:259 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 9 44 theme nonstandard 1310:1320 arg1 acid 1328:1331 this nonstandard amino acid 1305:1331 this nonstandard amino acid 1305:1331 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 9 45 theme biological 1285:1294 arg1 role 1296:1299 a biological role 1283:1299 a biological role for this nonstandard amino acid 1283:1331 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 10 46 theme GHs 1431:1433 arg1 interaction 1416:1426 the interaction 1412:1426 the interaction of GHs with their polymeric substrate 1412:1464 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 0 47 theme ring-size 24:32 arg1 determination 34:46 ring-size determination 24:46 ring-size determination 24:46 Amylose recognition and ring-size determination of amylomaltase. 28097217 6 48 with interaction 769:779 arg1 substrates 796:805 polymeric substrates 786:805 polymeric substrates that better represent the natural polysaccharide 786:854 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 6 49 theme polymeric 786:794 arg1 substrates 796:805 polymeric substrates 786:805 polymeric substrates that better represent the natural polysaccharide 786:854 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 5 50 theme intramolecular 504:517 arg1 cycloamyloses 597:609 the cycloamyloses 593:609 the cycloamyloses (CAs) 593:615 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 5 50 theme intramolecular 504:517 arg1 transglycosylation 519:536 an intramolecular transglycosylation 501:536 an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings 501:590 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 28097217 10 51 theme polymeric 1446:1454 arg1 substrate 1456:1464 their polymeric substrate 1440:1464 their polymeric substrate 1440:1464 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 0 52 theme amylomaltase 51:62 arg1 recognition 8:18 Amylose recognition 0:18 Amylose recognition 0:18 Amylose recognition and ring-size determination of amylomaltase. 28097217 0 52 theme amylomaltase 51:62 arg1 determination 34:46 ring-size determination 24:46 ring-size determination 24:46 Amylose recognition and ring-size determination of amylomaltase. 28097217 7 53 theme x-ray 997:1001 arg1 crystallography 1003:1017 x-ray crystallography 997:1017 x-ray crystallography 997:1017 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 2 54 theme linear 208:213 arg1 polymers 173:180 two carbohydrate polymers 156:180 two carbohydrate polymers 156:180 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 2 54 theme linear 208:213 arg1 amylose 215:221 linear amylose 208:221 linear amylose 208:221 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 6 55 theme short 685:689 arg1 binding 707:713 short oligosaccharide binding 685:713 short oligosaccharide binding to GHs 685:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 10 56 theme functional 1363:1372 arg1 assays 1374:1379 functional assays 1363:1379 functional assays 1363:1379 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 7 57 dep Thermus 901:907 arg1 aquaticus 909:917 aquaticus 909:917 aquaticus 909:917 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 4 58 link α-1,4-linked 432:443 arg1 chains 451:456 α-1,4-linked sugar chains 432:456 α-1,4-linked sugar chains such as amylose 432:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 4 58 link α-1,4-linked 432:443 arg1 amylose 466:472 amylose 466:472 amylose 466:472 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 7 59 theme carbohydrates 971:983 arg1 CA-one 949:954 a 34-meric CA-one 938:954 a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 938:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 10 60 with interaction 1416:1426 arg1 substrate 1456:1464 their polymeric substrate 1440:1464 their polymeric substrate 1440:1464 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 6 61 from binding 707:713 arg1 wealth 667:672 a wealth 665:672 a wealth of data on short oligosaccharide binding to GHs 665:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 2 62 from water 255:259 arg1 soluble 244:250 soluble 244:250 soluble 244:250 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 4 63 dep specialized 399:409 arg1 GHs 395:397 GHs 395:397 GHs 395:397 Amylomaltases (AMs) are GHs specialized in the hydrolysis of α-1,4-linked sugar chains such as amylose. 28097217 3 64 theme enzymatic 273:281 arg1 breakdown 283:291 the enzymatic breakdown 269:291 the enzymatic breakdown by glycoside hydrolases (GHs) 269:321 Hence, the enzymatic breakdown by glycoside hydrolases (GHs) is of great biological and societal importance. 28097217 2 65 theme branched 183:190 arg1 polymers 173:180 two carbohydrate polymers 156:180 two carbohydrate polymers 156:180 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 2 65 theme branched 183:190 arg1 amylopectin 192:202 branched amylopectin 183:202 branched amylopectin 183:202 It consists of two carbohydrate polymers, branched amylopectin and linear amylose, which are sparingly soluble in water. 28097217 6 66 theme data 677:680 arg1 wealth 667:672 a wealth 665:672 a wealth of data on short oligosaccharide binding to GHs 665:720 Despite a wealth of data on short oligosaccharide binding to GHs, no structural evidence is available for their interaction with polymeric substrates that better represent the natural polysaccharide. 28097217 7 67 from structure 888:896 arg1 complex 925:931 complex 925:931 complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 925:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 10 68 theme molecular 1480:1488 arg1 mechanism 1496:1504 a molecular ruler mechanism 1478:1504 a molecular ruler mechanism for minimal ring-size determination of CA products 1478:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 9 69 theme protein-sugar 1244:1256 arg1 interactions 1258:1269 protein-sugar interactions 1244:1269 protein-sugar interactions 1244:1269 A modified succinimide, derived from aspartate, mediates protein-sugar interactions, suggesting a biological role for this nonstandard amino acid. 28097217 7 70 theme 34-meric 940:947 arg1 CA-one 949:954 a 34-meric CA-one 938:954 a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 938:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 1 71 theme life 135:138 arg1 kingdoms 123:130 all kingdoms 119:130 all kingdoms of life 119:138 Starch is a major carbon and energy source throughout all kingdoms of life. 28097217 10 72 theme CA 1545:1546 arg1 products 1548:1555 CA products 1545:1555 CA products 1545:1555 The structure, together with functional assays, provides unique insights into the interaction of GHs with their polymeric substrate and reveals a molecular ruler mechanism for minimal ring-size determination of CA products. 28097217 7 73 with complex 925:931 arg1 CA-one 949:954 a 34-meric CA-one 938:954 a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate 938:1072 We have determined the crystal structure of Thermus aquaticus AM in complex with a 34-meric CA-one of the largest carbohydrates resolved by x-ray crystallography and a mimic of the natural polymeric amylose substrate. 28097217 5 74 dep 100 638:640 arg1 to 635:636 to 635:636 to 635:636 They are able to catalyze an intramolecular transglycosylation of a bound sugar chain yielding polymeric sugar rings, the cycloamyloses (CAs), consisting of 20 to 100 glucose units. 26306702 2 0 theme IMS-MS 355:360 arg1 analyses 362:369 IMS-MS analyses 355:369 IMS-MS analyses of underivatized carbohydrates 355:400 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 4 1 theme disaccharide 827:838 arg1 isomers 840:846 ten disaccharide isomers 823:846 ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose) 823:960 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 2 2 contain have 443:446 arg2 ccs 482:484 ccs 482:484 ccs 482:484 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 2 contain have 443:446 arg2 sections 472:479 similar collision cross sections 448:479 similar collision cross sections (ccs) 448:485 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 2 contain have 443:446 arg1 ions 438:441 most isobaric precursor ions 414:441 most isobaric precursor ions 414:441 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 1 3 theme collision-induced 166:182 arg1 CID 198:200 CID 198:200 CID 198:200 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 1 3 theme collision-induced 166:182 arg1 dissociation 184:195 collision-induced dissociation 166:195 collision-induced dissociation (CID) 166:201 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 4 4 theme associated-fragment 730:748 arg1 ions 750:753 precursor- and associated-fragment ions 715:753 precursor- and associated-fragment ions 715:753 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 3 5 theme ccs 501:503 arg1 values 505:510 ccs values 501:510 ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis 501:573 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 4 6 theme ccs 699:701 arg1 values 689:694 values 689:694 values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose) 689:1033 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 7 dep isomers 987:993 arg1 maltotriose 1022:1032 maltotriose 1022:1032 maltotriose 1022:1032 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 7 dep isomers 987:993 arg1 melezitose 1007:1016 melezitose 1007:1016 melezitose 1007:1016 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 7 dep isomers 987:993 arg1 raffinose 996:1004 raffinose 996:1004 raffinose 996:1004 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 7 dep isomers 987:993 arg1 isomers 987:993 three trisaccharide isomers 967:993 three trisaccharide isomers (raffinose, melezitose and maltotriose) 967:1033 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 2 8 theme precursor 428:436 arg1 ions 438:441 most isobaric precursor ions 414:441 most isobaric precursor ions 414:441 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 3 9 theme fragment 525:532 arg1 ions 534:537 isomeric fragment ions 516:537 isomeric fragment ions 516:537 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 4 10 dep isomers 780:786 arg1 isomers 780:786 three monosaccharide isomers 759:786 three monosaccharide isomers (glucose, galactose and fructose) 759:820 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 10 dep isomers 780:786 arg1 fructose 812:819 fructose 812:819 fructose 812:819 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 10 dep isomers 780:786 arg1 galactose 798:806 galactose 798:806 galactose 798:806 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 10 dep isomers 780:786 arg1 glucose 789:795 glucose 789:795 glucose 789:795 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 0 11 theme carbohydrate 14:25 arg1 isomers 27:33 carbohydrate isomers 14:33 carbohydrate isomers 14:33 Investigating carbohydrate isomers by IMS-CID-IMS-MS: precursor and fragment ion cross-sections. 26306702 3 12 theme isomeric 516:523 arg1 ions 534:537 isomeric fragment ions 516:537 isomeric fragment ions 516:537 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 1 13 theme Ion 97:99 arg1 techniques 123:132 Ion mobility spectrometry techniques 97:132 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) 97:228 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 2 14 theme carbohydrates 388:400 arg1 analyses 362:369 IMS-MS analyses 355:369 IMS-MS analyses of underivatized carbohydrates 355:400 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 15 theme collision 456:464 arg1 ccs 482:484 ccs 482:484 ccs 482:484 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 15 theme collision 456:464 arg1 sections 472:479 similar collision cross sections 448:479 similar collision cross sections (ccs) 448:485 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 3 16 from contrast 491:498 arg1 different 585:593 different 585:593 different 585:593 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 1 17 theme mobility 101:108 arg1 techniques 123:132 Ion mobility spectrometry techniques 97:132 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) 97:228 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 3 18 from different 585:593 arg1 contrast 491:498 contrast 491:498 contrast 491:498 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 2 19 theme underivatized 374:386 arg1 carbohydrates 388:400 underivatized carbohydrates 374:400 underivatized carbohydrates 374:400 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 20 theme similar 448:454 arg1 ccs 482:484 ccs 482:484 ccs 482:484 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 20 theme similar 448:454 arg1 sections 472:479 similar collision cross sections 448:479 similar collision cross sections (ccs) 448:485 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 1 21 theme singly-lithiated 272:287 arg1 isomers 302:308 singly-lithiated carbohydrate isomers 272:308 singly-lithiated carbohydrate isomers 272:308 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 4 22 from He 707:708 arg1 values 689:694 values 689:694 values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose) 689:1033 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 1 23 theme spectrometry 110:121 arg1 techniques 123:132 Ion mobility spectrometry techniques 97:132 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) 97:228 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 4 24 theme trisaccharide 973:985 arg1 maltotriose 1022:1032 maltotriose 1022:1032 maltotriose 1022:1032 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 24 theme trisaccharide 973:985 arg1 melezitose 1007:1016 melezitose 1007:1016 melezitose 1007:1016 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 24 theme trisaccharide 973:985 arg1 raffinose 996:1004 raffinose 996:1004 raffinose 996:1004 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 24 theme trisaccharide 973:985 arg1 isomers 987:993 three trisaccharide isomers 967:993 three trisaccharide isomers (raffinose, melezitose and maltotriose) 967:1033 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 1 25 theme carbohydrate 289:300 arg1 isomers 302:308 singly-lithiated carbohydrate isomers 272:308 singly-lithiated carbohydrate isomers 272:308 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 4 26 theme precursor- 715:724 arg1 ions 750:753 precursor- and associated-fragment ions 715:753 precursor- and associated-fragment ions 715:753 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 1 27 theme mass 207:210 arg1 MS 226:227 MS 226:227 MS 226:227 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 1 27 theme mass 207:210 arg1 spectrometry 212:223 mass spectrometry 207:223 mass spectrometry (MS) 207:228 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 1 28 theme isomers 302:308 arg1 structures 258:267 the structures 254:267 the structures of singly-lithiated carbohydrate isomers 254:308 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 2 29 theme isobaric 419:426 arg1 ions 438:441 most isobaric precursor ions 414:441 most isobaric precursor ions 414:441 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 5 30 theme differentiating 1077:1091 arg1 carbohydrates 1103:1115 differentiating precursor carbohydrates 1077:1115 differentiating precursor carbohydrates 1077:1115 These values are discussed as a means of differentiating precursor carbohydrates. 26306702 0 31 dep precursor 54:62 arg1 cross-sections 81:94 ion cross-sections 77:94 ion cross-sections 77:94 Investigating carbohydrate isomers by IMS-CID-IMS-MS: precursor and fragment ion cross-sections. 26306702 2 32 theme most 414:417 arg1 ions 438:441 most isobaric precursor ions 414:441 most isobaric precursor ions 414:441 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 4 33 dep isomers 840:846 arg1 trehalose 880:888 trehalose 880:888 trehalose 880:888 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 cellobiose 891:900 cellobiose 891:900 cellobiose 891:900 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 maltose 930:936 maltose 930:936 maltose 930:936 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 leucrose 858:865 leucrose 858:865 leucrose 858:865 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 lactose 939:945 lactose 939:945 lactose 939:945 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 palatinose 868:877 palatinose 868:877 palatinose 868:877 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 β-gentiobiose 903:915 β-gentiobiose 903:915 β-gentiobiose 903:915 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 isomaltose 918:927 isomaltose 918:927 isomaltose 918:927 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 melibiose 951:959 melibiose 951:959 melibiose 951:959 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 33 dep isomers 840:846 arg1 sucrose 849:855 sucrose 849:855 sucrose 849:855 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 5 34 theme precursor 1093:1101 arg1 carbohydrates 1103:1115 differentiating precursor carbohydrates 1077:1115 differentiating precursor carbohydrates 1077:1115 These values are discussed as a means of differentiating precursor carbohydrates. 26306702 2 35 theme cases 348:352 arg1 exception 320:328 the exception 316:328 the exception of some favorable cases 316:352 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 5 36 theme carbohydrates 1103:1115 arg1 means 1068:1072 a means 1066:1072 a means of differentiating precursor carbohydrates 1066:1115 These values are discussed as a means of differentiating precursor carbohydrates. 26306702 5 36 theme carbohydrates 1103:1115 arg1 values 1042:1047 These values 1036:1047 These values 1036:1047 These values are discussed as a means of differentiating precursor carbohydrates. 26306702 1 37 used used 234:237 arg2 techniques 123:132 Ion mobility spectrometry techniques 97:132 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) 97:228 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 2 38 theme favorable 338:346 arg1 cases 348:352 some favorable cases 333:352 some favorable cases 333:352 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 4 39 theme monosaccharide 765:778 arg1 isomers 780:786 three monosaccharide isomers 759:786 three monosaccharide isomers (glucose, galactose and fructose) 759:820 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 39 theme monosaccharide 765:778 arg1 fructose 812:819 fructose 812:819 fructose 812:819 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 39 theme monosaccharide 765:778 arg1 galactose 798:806 galactose 798:806 galactose 798:806 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 4 39 theme monosaccharide 765:778 arg1 glucose 789:795 glucose 789:795 glucose 789:795 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 3 40 theme IMS-CID-IMS-MS 551:564 arg1 analysis 566:573 IMS-CID-IMS-MS analysis 551:573 IMS-CID-IMS-MS analysis 551:573 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 1 41 dep techniques 123:132 arg1 IMS-IMS 143:149 IMS-IMS 143:149 IMS-IMS 143:149 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 1 41 dep techniques 123:132 arg1 IMS 135:137 IMS 135:137 IMS 135:137 Ion mobility spectrometry techniques (IMS and IMS-IMS) combined with collision-induced dissociation (CID) and mass spectrometry (MS) are used to investigate the structures of singly-lithiated carbohydrate isomers. 26306702 4 42 from ccs 699:701 arg1 He 707:708 He 707:708 He 707:708 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 26306702 0 43 theme ion 77:79 arg1 cross-sections 81:94 ion cross-sections 77:94 ion cross-sections 77:94 Investigating carbohydrate isomers by IMS-CID-IMS-MS: precursor and fragment ion cross-sections. 26306702 3 44 theme isomeric 658:665 arg1 precursors 667:676 the isomeric precursors 654:676 the isomeric precursors 654:676 In contrast, ccs values for isomeric fragment ions obtained by IMS-CID-IMS-MS analysis are often different, and thus appear to be useful as a means of distinguishing the isomeric precursors. 26306702 0 45 dep Investigating 0:12 arg1 fragment 68:75 fragment 68:75 fragment 68:75 Investigating carbohydrate isomers by IMS-CID-IMS-MS: precursor and fragment ion cross-sections. 26306702 0 45 dep Investigating 0:12 arg1 precursor 54:62 precursor 54:62 precursor 54:62 Investigating carbohydrate isomers by IMS-CID-IMS-MS: precursor and fragment ion cross-sections. 26306702 2 46 theme cross 466:470 arg1 ccs 482:484 ccs 482:484 ccs 482:484 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 2 46 theme cross 466:470 arg1 sections 472:479 similar collision cross sections 448:479 similar collision cross sections (ccs) 448:485 With the exception of some favorable cases, IMS-MS analyses of underivatized carbohydrates reveal that most isobaric precursor ions have similar collision cross sections (ccs). 26306702 4 47 from values 689:694 arg1 He 707:708 He 707:708 He 707:708 We report values of ccs (in He) for precursor- and associated-fragment ions for three monosaccharide isomers (glucose, galactose and fructose), ten disaccharide isomers (sucrose, leucrose, palatinose, trehalose, cellobiose, β-gentiobiose, isomaltose, maltose, lactose and melibiose), and three trisaccharide isomers (raffinose, melezitose and maltotriose). 25212784 3 0 theme bovine 990:995 arg1 FF 997:998 bovine FF 990:998 bovine FF 990:998 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 1 theme dairy 915:919 arg1 heifers 921:927 dairy heifers 915:927 dairy heifers 915:927 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 2 theme profile 320:326 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 2 theme profile 320:326 arg1 analysis 288:295 an initial analysis 277:295 an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 277:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 3 gly nonsialylated 1227:1239 arg1 glycans 1241:1247 nonsialylated glycans 1227:1247 nonsialylated glycans 1227:1247 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 4 from cows 943:946 arg1 preovulatory 862:873 preovulatory 862:873 preovulatory 862:873 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 3 5 theme hypersialylated 1025:1039 arg1 structures 1041:1050 biantennary hypersialylated structures 1013:1050 biantennary hypersialylated structures 1013:1050 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 6 from differences 589:599 arg1 profile 617:623 the N-glycan profile 604:623 the N-glycan profile 604:623 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 7 theme dominant 641:648 arg1 follicles 666:674 dominant and subordinate follicles 641:674 dominant and subordinate follicles 641:674 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 8 theme N-glycan 760:767 arg1 profile 769:775 the N-glycan profile 756:775 the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 756:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 9 from cows 715:718 arg1 FF 633:634 FF 633:634 FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows 633:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 10 theme initial 280:286 arg1 analysis 288:295 an initial analysis 277:295 an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 277:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 11 from heifers 921:927 arg1 preovulatory 862:873 preovulatory 862:873 preovulatory 862:873 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 1 12 theme many 187:190 arg1 proteins 192:199 many proteins 187:199 many proteins that are glycosylated with N-linked glycans 187:243 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 5 13 from structures 1473:1482 arg1 N-glycans 1490:1498 FF N-glycans 1487:1498 FF N-glycans 1487:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 2 14 from alterations 741:751 arg1 profile 769:775 the N-glycan profile 756:775 the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 756:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 15 theme dairy 681:685 arg1 heifers 687:693 dairy heifers 681:693 dairy heifers 681:693 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 16 theme dairy 709:713 arg1 cows 715:718 lactating dairy cows 699:718 lactating dairy cows 699:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 17 theme higher 1210:1215 arg1 levels 1217:1222 higher levels 1210:1222 higher levels of nonsialylated glycans 1210:1247 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 18 theme N-glycans 1168:1176 arg1 comparison 1151:1160 A comparison 1149:1160 A comparison of FF N-glycans from cows and heifers 1149:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 19 from heifers 687:693 arg1 FF 633:634 FF 633:634 FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows 633:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 20 theme dominant 53:60 arg1 stages 85:90 key dominant follicle developmental stages 49:90 key dominant follicle developmental stages 49:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 2 21 theme -based 470:475 arg1 separations 477:487 -based separations 470:487 -based separations 470:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 22 theme developmental 71:83 arg1 stages 85:90 key dominant follicle developmental stages 49:90 key dominant follicle developmental stages 49:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 1 23 gly glycosylated 210:221 arg1 proteins 192:199 many proteins 187:199 many proteins that are glycosylated with N-linked glycans 187:243 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 2 24 theme spectrometry/mass 531:547 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 25 from proportion 1262:1271 arg1 cows 1298:1301 cows 1298:1301 cows 1298:1301 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 26 theme liquid 504:509 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 5 27 theme sialylated 1462:1471 arg1 structures 1473:1482 sialylated structures 1462:1482 sialylated structures on FF N-glycans 1462:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 5 28 gly sialylated 1462:1471 arg1 structures 1473:1482 sialylated structures 1462:1482 sialylated structures on FF N-glycans 1462:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 2 29 theme liquid 371:376 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 29 theme liquid 371:376 arg1 chromatography 409:422 anion exchange chromatography 394:422 anion exchange chromatography 394:422 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 29 theme liquid 371:376 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 30 theme performance 430:440 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 30 theme performance 430:440 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 30 theme performance 430:440 arg1 HPLC 465:468 HPLC 465:468 HPLC 465:468 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 31 theme hydrophilic 347:357 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 31 theme hydrophilic 347:357 arg1 chromatography 409:422 anion exchange chromatography 394:422 anion exchange chromatography 394:422 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 31 theme hydrophilic 347:357 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 32 theme N-glycan 0:7 arg1 profiling 9:17 N-glycan profiling 0:17 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages 0:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 1 33 theme Follicular 93:102 arg1 microenvironment 129:144 an important microenvironment 116:144 an important microenvironment for the development of oocytes 116:175 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 33 theme Follicular 93:102 arg1 FF 111:112 FF 111:112 FF 111:112 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 33 theme Follicular 93:102 arg1 fluid 104:108 Follicular fluid 93:108 Follicular fluid (FF) 93:113 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 2 34 theme lactating 933:941 arg1 cows 943:946 lactating cows 933:946 lactating cows 933:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 35 theme glycans 1241:1247 arg1 levels 1217:1222 higher levels 1210:1222 higher levels of nonsialylated glycans 1210:1247 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 36 theme exchange 400:407 arg1 chromatography 409:422 anion exchange chromatography 394:422 anion exchange chromatography 394:422 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 36 theme exchange 400:407 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 3 37 from majority 967:974 arg1 FF 997:998 bovine FF 990:998 bovine FF 990:998 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 0 38 theme follicular 29:38 arg1 fluid 40:44 bovine follicular fluid 22:44 bovine follicular fluid 22:44 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 3 39 theme terminal 1132:1139 arg1 sugars 1141:1146 terminal sugars 1132:1146 terminal sugars 1132:1146 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 3 40 from glycans 979:985 arg1 FF 997:998 bovine FF 990:998 bovine FF 990:998 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 41 theme glycan 313:318 arg1 profile 320:326 the N-linked glycan profile 300:326 the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 300:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 42 from heifers 1192:1198 arg1 comparison 1151:1160 A comparison 1149:1160 A comparison of FF N-glycans from cows and heifers 1149:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 42 from heifers 1192:1198 arg1 N-glycans 1168:1176 FF N-glycans 1165:1176 FF N-glycans from cows and heifers 1165:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 43 theme sialylated 1276:1285 arg1 glycans 1287:1293 sialylated glycans 1276:1293 sialylated glycans 1276:1293 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 44 from cows 1183:1186 arg1 comparison 1151:1160 A comparison 1149:1160 A comparison of FF N-glycans from cows and heifers 1149:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 44 from cows 1183:1186 arg1 N-glycans 1168:1176 FF N-glycans 1165:1176 FF N-glycans from cows and heifers 1165:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 1 45 contain contains 178:185 arg1 microenvironment 129:144 an important microenvironment 116:144 an important microenvironment for the development of oocytes 116:175 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 45 contain contains 178:185 arg1 FF 111:112 FF 111:112 FF 111:112 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 45 contain contains 178:185 arg2 proteins 192:199 many proteins 187:199 many proteins that are glycosylated with N-linked glycans 187:243 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 45 contain contains 178:185 arg1 fluid 104:108 Follicular fluid 93:108 Follicular fluid (FF) 93:113 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 2 46 theme FF 338:339 arg1 profile 320:326 the N-linked glycan profile 300:326 the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 300:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 3 47 dep galactose 1098:1106 arg1 sugars 1141:1146 terminal sugars 1132:1146 terminal sugars 1132:1146 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 3 47 dep galactose 1098:1106 arg1 the 1094:1096 the 1094:1096 the 1094:1096 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 3 48 gly hypersialylated 1025:1039 arg1 structures 1041:1050 biantennary hypersialylated structures 1013:1050 biantennary hypersialylated structures 1013:1050 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 49 theme N-glycan 608:615 arg1 profile 617:623 the N-glycan profile 604:623 the N-glycan profile 604:623 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 3 50 theme biantennary 1013:1023 arg1 structures 1041:1050 biantennary hypersialylated structures 1013:1050 biantennary hypersialylated structures 1013:1050 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 51 link N-linked 304:311 arg1 profile 320:326 the N-linked glycan profile 300:326 the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 300:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 5 52 theme overall 1442:1448 arg1 decrease 1450:1457 an overall decrease 1439:1457 an overall decrease in sialylated structures on FF N-glycans 1439:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 2 53 theme dominant 891:898 arg1 follicles 900:908 newly selected, differentiated (preovulatory) and luteinised dominant follicles 830:908 follicles 900:908 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 54 from comparison 1151:1160 arg1 cows 1183:1186 cows 1183:1186 cows 1183:1186 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 54 from comparison 1151:1160 arg1 heifers 1192:1198 heifers 1192:1198 heifers 1192:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 55 theme FF 1165:1166 arg1 N-glycans 1168:1176 FF N-glycans 1165:1176 FF N-glycans from cows and heifers 1165:1198 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 1 56 link N-linked 228:235 arg1 glycans 237:243 N-linked glycans 228:243 N-linked glycans 228:243 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 57 theme oocytes 169:175 arg1 development 154:164 the development 150:164 the development of oocytes 150:175 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 2 58 theme preovulatory 790:801 arg1 development 812:822 preovulatory follicle development 790:822 preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 790:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 59 from stages 85:90 arg1 profiling 9:17 N-glycan profiling 0:17 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages 0:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 2 60 theme FF 780:781 arg1 profile 769:775 the N-glycan profile 756:775 the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 756:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 61 theme spectrometry 549:560 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 5 62 from N-glycans 1490:1498 arg1 decrease 1450:1457 an overall decrease 1439:1457 an overall decrease in sialylated structures on FF N-glycans 1439:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 2 63 dep present 269:275 arg1 identify 732:739 identify 732:739 iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 724:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 63 dep present 269:275 arg1 ii 572:573 ii 572:573 ii 572:573 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 63 dep present 269:275 arg1 determine 579:587 determine 579:587 to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows 576:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 63 dep present 269:275 arg1 i 263:263 i 263:263 i 263:263 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 64 from follicles 666:674 arg1 FF 633:634 FF 633:634 FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows 633:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 65 theme follicle 62:69 arg1 stages 85:90 key dominant follicle developmental stages 49:90 key dominant follicle developmental stages 49:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 1 66 theme important 119:127 arg1 microenvironment 129:144 an important microenvironment 116:144 an important microenvironment for the development of oocytes 116:175 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 1 66 theme important 119:127 arg1 fluid 104:108 Follicular fluid 93:108 Follicular fluid (FF) 93:113 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 25212784 4 67 gly sialylated 1276:1285 arg1 glycans 1287:1293 sialylated glycans 1276:1293 sialylated glycans 1276:1293 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 4 68 theme lower 1256:1260 arg1 proportion 1262:1271 a lower proportion 1254:1271 a lower proportion of sialylated glycans in cows 1254:1301 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 69 theme chromatography-mass 511:529 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 70 theme subordinate 654:664 arg1 follicles 666:674 dominant and subordinate follicles 641:674 dominant and subordinate follicles 641:674 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 71 theme subsequent 493:502 arg1 analysis 562:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis 493:569 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 72 theme lactating 699:707 arg1 cows 715:718 lactating dairy cows 699:718 lactating dairy cows 699:718 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 73 theme interaction 359:369 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 73 theme interaction 359:369 arg1 chromatography 409:422 anion exchange chromatography 394:422 anion exchange chromatography 394:422 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 73 theme interaction 359:369 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 74 dep identify 732:739 arg1 iii 724:726 iii 724:726 iii 724:726 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 3 75 from FF 997:998 arg1 majority 967:974 the majority 963:974 the majority of glycans on bovine FF 963:998 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 76 theme high 425:428 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 76 theme high 425:428 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 76 theme high 425:428 arg1 HPLC 465:468 HPLC 465:468 HPLC 465:468 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 5 77 theme FF 1487:1488 arg1 N-glycans 1490:1498 FF N-glycans 1487:1498 FF N-glycans 1487:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 0 78 theme bovine 22:27 arg1 fluid 40:44 bovine follicular fluid 22:44 bovine follicular fluid 22:44 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 2 79 theme anion 394:398 arg1 chromatography 409:422 anion exchange chromatography 394:422 anion exchange chromatography 394:422 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 79 theme anion 394:398 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 80 theme fluid 40:44 arg1 profiling 9:17 N-glycan profiling 0:17 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages 0:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 3 81 gly sialylated 1075:1084 arg1 glycans 1063:1069 the glycans 1059:1069 the glycans 1059:1069 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 82 dep chromatography 378:391 arg1 separations 477:487 -based separations 470:487 -based separations 470:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 0 83 theme key 49:51 arg1 stages 85:90 key dominant follicle developmental stages 49:90 key dominant follicle developmental stages 49:90 N-glycan profiling of bovine follicular fluid at key dominant follicle developmental stages. 25212784 3 84 theme glycans 979:985 arg1 majority 967:974 the majority 963:974 the majority of glycans on bovine FF 963:998 We found that the majority of glycans on bovine FF are based on biantennary hypersialylated structures, where the glycans are sialylated on both the galactose and N-acetylglucosamine terminal sugars. 25212784 2 85 theme liquid 442:447 arg1 chromatography 449:462 high performance liquid chromatography 425:462 high performance liquid chromatography (HPLC) 425:469 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 85 theme liquid 442:447 arg1 chromatography 378:391 hydrophilic interaction liquid chromatography 347:391 hydrophilic interaction liquid chromatography 347:391 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 85 theme liquid 442:447 arg1 HPLC 465:468 HPLC 465:468 HPLC 465:468 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 86 theme glycans 1287:1293 arg1 proportion 1262:1271 a lower proportion 1254:1271 a lower proportion of sialylated glycans in cows 1254:1301 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 87 theme N-linked 304:311 arg1 profile 320:326 the N-linked glycan profile 300:326 the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 300:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 4 88 theme nonsialylated 1227:1239 arg1 glycans 1241:1247 nonsialylated glycans 1227:1247 nonsialylated glycans 1227:1247 A comparison of FF N-glycans from cows and heifers indicated higher levels of nonsialylated glycans with a lower proportion of sialylated glycans in cows than in heifers. 25212784 2 89 theme luteinised 880:889 arg1 follicles 900:908 newly selected, differentiated (preovulatory) and luteinised dominant follicles 830:908 follicles 900:908 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 5 90 from decrease 1450:1457 arg1 structures 1473:1482 sialylated structures 1462:1482 sialylated structures on FF N-glycans 1462:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 5 90 from decrease 1450:1457 arg1 N-glycans 1490:1498 FF N-glycans 1487:1498 FF N-glycans 1487:1498 Overall, as the follicle develops from Selection, Differentiation and Luteinisation in both cows and heifers, there is an overall decrease in sialylated structures on FF N-glycans. 25212784 2 91 theme follicle 803:810 arg1 development 812:822 preovulatory follicle development 790:822 preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows 790:946 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 2 92 theme bovine 331:336 arg1 FF 338:339 bovine FF 331:339 bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations 331:487 This study aimed i) to present an initial analysis of the N-linked glycan profile of bovine FF using hydrophilic interaction liquid chromatography, anion exchange chromatography, high performance liquid chromatography (HPLC)-based separations and subsequent liquid chromatography-mass spectrometry/mass spectrometry analysis; ii) to determine differences in the N-glycan profile between FF from dominant and subordinate follicles from dairy heifers and lactating dairy cows and iii) to identify alterations in the N-glycan profile of FF during preovulatory follicle development using newly selected, differentiated (preovulatory) and luteinised dominant follicles from dairy heifers and lactating cows. 25212784 1 93 theme N-linked 228:235 arg1 glycans 237:243 N-linked glycans 228:243 N-linked glycans 228:243 Follicular fluid (FF), an important microenvironment for the development of oocytes, contains many proteins that are glycosylated with N-linked glycans. 28424265 8 0 theme GNE-deficient 1151:1163 arg1 cells 1165:1169 GNE-deficient cells 1151:1169 GNE-deficient cells 1151:1169 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 10 1 theme N-linked 1595:1602 arg1 branching 1611:1619 N-linked glycan branching 1595:1619 N-linked glycan branching 1595:1619 Furthermore, GNE deficiency and glucose supplementation acted independently and additively to increase N-linked glycan branching. 28424265 5 2 theme sialic 642:647 arg1 production 654:663 sialic acid production 642:663 sialic acid production 642:663 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 2 3 theme sporadic 328:335 arg1 myositis 352:359 sporadic inclusion body myositis 328:359 sporadic inclusion body myositis 328:359 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 9 4 link N-linked 1465:1472 arg1 structure 1481:1489 N-linked glycan structure 1465:1489 N-linked glycan structure 1465:1489 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 11 5 theme enhanced 1690:1697 arg1 binding 1699:1705 enhanced binding 1690:1705 enhanced binding to galectin-1 1690:1719 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 4 6 theme GNE 565:567 arg1 myopathy 569:576 GNE myopathy 565:576 GNE myopathy 565:576 Mutations to both GNE domains are linked to GNE myopathy. 28424265 11 7 theme GNE 1749:1751 arg1 activity 1753:1760 GNE activity 1749:1760 GNE activity 1749:1760 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 6 8 theme cell 795:798 arg1 lines 800:804 cell lines 795:804 cell lines expressing wild type or mutant forms of GNE 795:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 9 9 theme N-linked 1465:1472 arg1 structure 1481:1489 N-linked glycan structure 1465:1489 N-linked glycan structure 1465:1489 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 8 10 theme N-linked 1189:1196 arg1 structures 1205:1214 distinct N-linked glycan structures 1180:1214 distinct N-linked glycan structures 1180:1214 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 11 11 theme N-linked 1631:1638 arg1 glycans 1640:1646 N-linked glycans 1631:1646 N-linked glycans produced by GNE-deficient cells 1631:1678 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 1 12 theme muscle 166:171 arg1 disorder 173:180 a rare muscle disorder 159:180 a rare muscle disorder associated with aging 159:202 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 12 theme muscle 166:171 arg1 myopathy 147:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 6 13 theme sialic 769:774 arg1 acid 776:779 sialic acid 769:779 sialic acid production in cell lines expressing wild type or mutant forms of GNE 769:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 8 14 theme increased 1221:1229 arg1 branching 1231:1239 increased branching 1221:1239 increased branching 1221:1239 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 9 15 theme glycan 1474:1479 arg1 structure 1481:1489 N-linked glycan structure 1465:1489 N-linked glycan structure 1465:1489 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 7 16 theme cell-surface 1076:1087 arg1 glycans 1089:1095 cell-surface glycans 1076:1095 cell-surface glycans 1076:1095 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 3 17 theme sialic 489:494 arg1 acid 496:499 sialic acid 489:499 sialic acid 489:499 GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. 28424265 5 18 from reductions 628:637 arg1 severity 677:684 disease severity 669:684 disease severity 669:684 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 5 18 from reductions 628:637 arg1 production 654:663 sialic acid production 642:663 sialic acid production 642:663 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 5 19 theme disease 669:675 arg1 severity 677:684 disease severity 669:684 disease severity 669:684 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 7 20 theme GNE 1000:1002 arg1 deficiency 1004:1013 GNE deficiency 1000:1013 GNE deficiency 1000:1013 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 1 21 theme UDP-GlcNAc 109:118 arg1 kinase 139:144 UDP-GlcNAc 2-epimerase/ManNAc kinase 109:144 UDP-GlcNAc 2-epimerase/ManNAc kinase 109:144 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 21 theme UDP-GlcNAc 109:118 arg1 GNE 104:106 GNE 104:106 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 11 22 gly glycoproteins 1797:1809 arg1 glycoproteins 1797:1809 cell-surface glycoproteins 1784:1809 cell-surface glycoproteins 1784:1809 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 7 23 theme unanticipated 1034:1046 arg1 effects 1048:1054 unanticipated effects 1034:1054 unanticipated effects on the structure of cell-surface glycans 1034:1095 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 4 24 attach linked 555:560 arg2 Mutations 521:529 Mutations 521:529 Mutations to both GNE domains 521:549 Mutations to both GNE domains are linked to GNE myopathy. 28424265 4 24 attach linked 555:560 arg1 myopathy 569:576 GNE myopathy 565:576 GNE myopathy 565:576 Mutations to both GNE domains are linked to GNE myopathy. 28424265 8 25 link N-linked 1189:1196 arg1 structures 1205:1214 distinct N-linked glycan structures 1180:1214 distinct N-linked glycan structures 1180:1214 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 11 26 theme galectin 1819:1826 arg1 lattice 1828:1834 the galectin lattice 1815:1834 the galectin lattice 1815:1834 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 0 27 theme glycan 56:61 arg1 branching 63:71 N-linked glycan branching 47:71 N-linked glycan branching 47:71 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 7 28 theme spectrometry 964:975 arg1 analysis 977:984 mass spectrometry analysis 959:984 mass spectrometry analysis 959:984 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 2 29 from mutations 405:413 arg1 GNE 418:420 GNE 418:420 GNE 418:420 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 9 30 theme key 1330:1332 arg1 metabolite 1334:1343 a key metabolite 1328:1343 a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway 1328:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 9 30 theme key 1330:1332 arg1 UDP-GlcNAc 1316:1325 UDP-GlcNAc 1316:1325 UDP-GlcNAc 1316:1325 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 0 31 from Effects 0:6 arg1 interactions 90:101 cell surface interactions 77:101 cell surface interactions 77:101 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 0 31 from Effects 0:6 arg1 branching 63:71 N-linked glycan branching 47:71 N-linked glycan branching 47:71 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 0 32 theme surface 82:88 arg1 interactions 90:101 cell surface interactions 77:101 cell surface interactions 77:101 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 7 33 theme lectin 940:945 arg1 binding 947:953 lectin binding 940:953 lectin binding 940:953 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 2 34 theme GNE 373:375 arg1 myopathy 377:384 GNE myopathy 373:384 GNE myopathy 373:384 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 9 35 theme nutrient-sensing 1352:1367 arg1 pathway 1393:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 11 36 from changes 1738:1744 arg1 activity 1753:1760 GNE activity 1749:1760 GNE activity 1749:1760 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 10 37 theme GNE 1505:1507 arg1 deficiency 1509:1518 GNE deficiency 1505:1518 GNE deficiency 1505:1518 Furthermore, GNE deficiency and glucose supplementation acted independently and additively to increase N-linked glycan branching. 28424265 9 38 theme biosynthetic 1380:1391 arg1 pathway 1393:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 0 39 theme altered 11:17 arg1 biosynthesis 31:42 altered sialic acid biosynthesis 11:42 altered sialic acid biosynthesis 11:42 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 1 40 theme acquired 272:279 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 40 theme acquired 272:279 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 41 theme GNE 104:106 arg1 disorder 173:180 a rare muscle disorder 159:180 a rare muscle disorder associated with aging 159:202 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 41 theme GNE 104:106 arg1 myopathy 147:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 4 42 theme GNE 539:541 arg1 domains 543:549 both GNE domains 534:549 both GNE domains 534:549 Mutations to both GNE domains are linked to GNE myopathy. 28424265 0 43 theme acid 26:29 arg1 biosynthesis 31:42 altered sialic acid biosynthesis 11:42 altered sialic acid biosynthesis 11:42 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 9 44 from change 1455:1460 arg1 structure 1481:1489 N-linked glycan structure 1465:1489 N-linked glycan structure 1465:1489 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 10 45 link N-linked 1595:1602 arg1 branching 1611:1619 N-linked glycan branching 1595:1619 N-linked glycan branching 1595:1619 Furthermore, GNE deficiency and glucose supplementation acted independently and additively to increase N-linked glycan branching. 28424265 1 46 theme aging 299:303 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 46 theme aging 299:303 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 6 47 theme mutant 830:835 arg1 forms 837:841 mutant forms 830:841 mutant forms of GNE 830:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 6 48 theme GNE 742:744 arg1 mutations 746:754 GNE mutations 742:754 GNE mutations 742:754 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 7 49 theme attributable 894:905 arg1 differences 882:892 any differences 878:892 any differences attributable to disease-associated mutations 878:937 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 8 50 theme low 1124:1126 arg1 levels 1128:1133 low levels 1124:1133 low levels of sialylation 1124:1148 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 11 51 theme glycoproteins 1797:1809 arg1 affinity 1772:1779 affinity 1772:1779 affinity of cell-surface glycoproteins for the galectin lattice 1772:1834 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 3 52 theme acid 496:499 arg1 biosynthesis 473:484 biosynthesis 473:484 biosynthesis of sialic acid in mammalian cells 473:518 GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. 28424265 1 53 theme sporadic 222:229 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 53 theme sporadic 222:229 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 12 54 theme GNE 1896:1898 arg1 activity 1900:1907 GNE activity 1896:1907 GNE activity 1896:1907 These findings suggest an unanticipated mechanism by which GNE activity might affect signaling through cell-surface receptors. 28424265 10 55 theme glycan 1604:1609 arg1 branching 1611:1619 N-linked glycan branching 1595:1619 N-linked glycan branching 1595:1619 Furthermore, GNE deficiency and glucose supplementation acted independently and additively to increase N-linked glycan branching. 28424265 6 56 theme other 715:719 arg1 effects 731:737 other potential effects 715:737 other potential effects of GNE mutations 715:754 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 2 57 theme inclusion 337:345 arg1 myositis 352:359 sporadic inclusion body myositis 328:359 sporadic inclusion body myositis 328:359 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 1 58 theme body 241:244 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 58 theme body 241:244 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 7 59 theme glycans 1089:1095 arg1 structure 1063:1071 the structure 1059:1071 the structure of cell-surface glycans 1059:1095 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 5 60 theme acid 649:652 arg1 production 654:663 sialic acid production 642:663 sialic acid production 642:663 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 12 61 theme unanticipated 1863:1875 arg1 mechanism 1877:1885 an unanticipated mechanism 1860:1885 an unanticipated mechanism by which GNE activity might affect signaling through cell-surface receptors 1860:1961 These findings suggest an unanticipated mechanism by which GNE activity might affect signaling through cell-surface receptors. 28424265 3 62 theme mammalian 504:512 arg1 cells 514:518 mammalian cells 504:518 mammalian cells 504:518 GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. 28424265 1 63 theme rare 161:164 arg1 disorder 173:180 a rare muscle disorder 159:180 a rare muscle disorder associated with aging 159:202 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 63 theme rare 161:164 arg1 myopathy 147:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 8 64 theme distinct 1180:1187 arg1 structures 1205:1214 distinct N-linked glycan structures 1180:1214 distinct N-linked glycan structures 1180:1214 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 0 65 link N-linked 47:54 arg1 branching 63:71 N-linked glycan branching 47:71 N-linked glycan branching 47:71 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 5 66 theme mutation-associated 608:626 arg1 reductions 628:637 mutation-associated reductions 608:637 mutation-associated reductions in sialic acid production and disease severity 608:684 However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect. 28424265 6 67 theme acid 776:779 arg1 production 781:790 sialic acid production 769:790 sialic acid production in cell lines expressing wild type or mutant forms of GNE 769:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 8 68 theme glycan 1198:1203 arg1 structures 1205:1214 distinct N-linked glycan structures 1180:1214 distinct N-linked glycan structures 1180:1214 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 8 69 dep exhibiting 1113:1122 arg1 addition 1101:1108 addition 1101:1108 addition 1101:1108 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 3 70 from biosynthesis 473:484 arg1 cells 514:518 mammalian cells 504:518 mammalian cells 504:518 GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. 28424265 2 71 theme myositis 352:359 arg1 cause 319:323 the cause 315:323 the cause of sporadic inclusion body myositis 315:359 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 2 71 theme myositis 352:359 arg1 unknown 364:370 unknown 364:370 unknown 364:370 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 9 72 theme GNE 1280:1282 arg1 deficiency 1284:1293 GNE deficiency 1280:1293 GNE deficiency 1280:1293 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 9 73 from metabolite 1334:1343 arg1 pathway 1393:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 8 74 theme extended 1245:1252 arg1 poly-N-acetyllactosamine 1254:1277 extended poly-N-acetyllactosamine 1245:1277 extended poly-N-acetyllactosamine 1245:1277 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 1 75 theme 2-epimerase/ManNAc 120:137 arg1 kinase 139:144 UDP-GlcNAc 2-epimerase/ManNAc kinase 109:144 UDP-GlcNAc 2-epimerase/ManNAc kinase 109:144 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 75 theme 2-epimerase/ManNAc 120:137 arg1 GNE 104:106 GNE 104:106 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy 104:154 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 9 76 theme UDP-GlcNAc 1316:1325 arg1 levels 1306:1311 levels 1306:1311 levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway 1306:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 0 77 theme cell 77:80 arg1 interactions 90:101 cell surface interactions 77:101 cell surface interactions 77:101 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 7 78 theme mass 959:962 arg1 analysis 977:984 mass spectrometry analysis 959:984 mass spectrometry analysis 959:984 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 9 79 theme hexosamine 1369:1378 arg1 pathway 1393:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 the nutrient-sensing hexosamine biosynthetic pathway 1348:1399 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 0 80 theme sialic 19:24 arg1 biosynthesis 31:42 altered sialic acid biosynthesis 11:42 altered sialic acid biosynthesis 11:42 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 1 81 theme common 265:270 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 81 theme common 265:270 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 7 82 theme disease-associated 910:927 arg1 mutations 929:937 disease-associated mutations 910:937 disease-associated mutations 910:937 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 3 83 theme enzymatic 439:447 arg1 activities 449:458 two enzymatic activities 435:458 two enzymatic activities required for biosynthesis of sialic acid in mammalian cells 435:518 GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. 28424265 0 84 theme biosynthesis 31:42 arg1 Effects 0:6 Effects 0:6 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 0:102 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 1 85 theme muscle 281:286 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 85 theme muscle 281:286 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 6 86 theme GNE 846:848 arg1 type 822:825 wild type 817:825 wild type 817:825 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 6 86 theme GNE 846:848 arg1 forms 837:841 mutant forms 830:841 mutant forms of GNE 830:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 6 87 from production 781:790 arg1 lines 800:804 cell lines 795:804 cell lines expressing wild type or mutant forms of GNE 795:848 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 10 88 theme glucose 1524:1530 arg1 supplementation 1532:1546 glucose supplementation 1524:1546 glucose supplementation 1524:1546 Furthermore, GNE deficiency and glucose supplementation acted independently and additively to increase N-linked glycan branching. 28424265 11 89 link N-linked 1631:1638 arg1 glycans 1640:1646 N-linked glycans 1631:1646 N-linked glycans produced by GNE-deficient cells 1631:1678 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 11 90 theme GNE-deficient 1660:1672 arg1 cells 1674:1678 GNE-deficient cells 1660:1678 GNE-deficient cells 1660:1678 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 0 91 theme N-linked 47:54 arg1 branching 63:71 N-linked glycan branching 47:71 N-linked glycan branching 47:71 Effects of altered sialic acid biosynthesis on N-linked glycan branching and cell surface interactions. 28424265 9 92 theme modest 1411:1416 arg1 effect 1418:1423 this modest effect 1406:1423 this modest effect 1406:1423 GNE deficiency may affect levels of UDP-GlcNAc, a key metabolite in the nutrient-sensing hexosamine biosynthetic pathway, but this modest effect did not fully account for the change in N-linked glycan structure. 28424265 12 93 theme cell-surface 1940:1951 arg1 receptors 1953:1961 cell-surface receptors 1940:1961 cell-surface receptors 1940:1961 These findings suggest an unanticipated mechanism by which GNE activity might affect signaling through cell-surface receptors. 28424265 6 94 theme mutations 746:754 arg1 effects 731:737 other potential effects 715:737 other potential effects of GNE mutations 715:754 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 11 95 theme cell-surface 1784:1795 arg1 glycoproteins 1797:1809 cell-surface glycoproteins 1784:1809 cell-surface glycoproteins 1784:1809 Notably, N-linked glycans produced by GNE-deficient cells displayed enhanced binding to galectin-1, indicating that changes in GNE activity can alter affinity of cell-surface glycoproteins for the galectin lattice. 28424265 8 96 theme sialylation 1138:1148 arg1 levels 1128:1133 low levels 1124:1133 low levels of sialylation 1124:1148 In addition to exhibiting low levels of sialylation, GNE-deficient cells produced distinct N-linked glycan structures with increased branching and extended poly-N-acetyllactosamine. 28424265 6 97 theme wild 817:820 arg1 type 822:825 wild type 817:825 wild type 817:825 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 7 98 from effects 1048:1054 arg1 structure 1063:1071 the structure 1059:1071 the structure of cell-surface glycans 1059:1095 Although we did not detect any differences attributable to disease-associated mutations, lectin binding and mass spectrometry analysis revealed that GNE deficiency is associated with unanticipated effects on the structure of cell-surface glycans. 28424265 6 99 theme potential 721:729 arg1 effects 731:737 other potential effects 715:737 other potential effects of GNE mutations 715:754 To investigate other potential effects of GNE mutations, we compared sialic acid production in cell lines expressing wild type or mutant forms of GNE. 28424265 2 100 theme body 347:350 arg1 myositis 352:359 sporadic inclusion body myositis 328:359 sporadic inclusion body myositis 328:359 Although the cause of sporadic inclusion body myositis is unknown, GNE myopathy is associated with mutations in GNE. 28424265 1 101 theme inclusion 231:239 arg1 myositis 246:253 sporadic inclusion body myositis 222:253 sporadic inclusion body myositis 222:253 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 28424265 1 101 theme inclusion 231:239 arg1 disease 288:294 the most common acquired muscle disease 256:294 the most common acquired muscle disease of aging 256:303 GNE (UDP-GlcNAc 2-epimerase/ManNAc kinase) myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis, the most common acquired muscle disease of aging. 25818946 6 0 theme bacterial 1333:1341 arg1 lattices 1364:1371 bacterial and archaeal S-layer lattices 1333:1371 bacterial and archaeal S-layer lattices 1333:1371 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 6 1 theme latter 1263:1268 arg1 features 1270:1277 The latter features 1259:1277 The latter features 1259:1277 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 6 1 theme latter 1263:1268 arg1 ones 1322:1325 the most general ones 1305:1325 the most general ones 1305:1325 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 6 1 theme latter 1263:1268 arg1 one 1298:1300 one 1298:1300 one 1298:1300 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 5 2 theme present 1076:1082 arg1 results 1084:1090 The present results 1072:1090 The present results 1072:1090 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 1 3 theme antifouling 264:274 arg1 property 276:283 accompanied antifouling property 252:283 accompanied antifouling property 252:283 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 4 theme nowadays 339:346 arg1 task 369:372 nowadays a highly challenging task 339:372 nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences 339:451 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 5 theme bio 431:433 arg1 sciences 444:451 (bio)material sciences 430:451 (bio)material sciences 430:451 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 4 6 theme non-glycosylated 986:1001 arg1 rSgsE 1003:1007 the non-glycosylated rSgsE 982:1007 the non-glycosylated rSgsE with the preferred region being the extending glycan residues 982:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 4 7 theme extending 1045:1053 arg1 residues 1062:1069 the extending glycan residues 1041:1069 the extending glycan residues 1041:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 6 8 theme ones 1322:1325 arg1 features 1270:1277 The latter features 1259:1277 The latter features 1259:1277 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 6 8 theme ones 1322:1325 arg1 ones 1322:1325 the most general ones 1305:1325 the most general ones 1305:1325 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 6 8 theme ones 1322:1325 arg1 one 1298:1300 one 1298:1300 one 1298:1300 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 5 9 from effect 1122:1127 arg1 behavior 1249:1256 fluidic behavior 1241:1256 fluidic behavior 1241:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 9 from effect 1122:1127 arg1 roughness 1227:1235 the nanoscale roughness 1213:1235 the nanoscale roughness 1213:1235 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 9 from effect 1122:1127 arg1 layer 1202:1206 the adjoining hydration layer 1178:1206 the adjoining hydration layer 1178:1206 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 9 from effect 1122:1127 arg1 recrystallization 1159:1175 the recrystallization 1155:1175 the recrystallization 1155:1175 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 3 10 dep soft 838:841 arg1 dissipative 851:861 dissipative 851:861 dissipative 851:861 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 5 11 theme fluidic 1241:1247 arg1 behavior 1249:1256 fluidic behavior 1241:1256 fluidic behavior 1241:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 2 12 from stearothermophilus 571:588 arg1 recrystallization 491:507 the recrystallization 487:507 the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus 487:588 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 6 13 theme general 1314:1320 arg1 ones 1322:1325 the most general ones 1305:1325 the most general ones 1305:1325 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 3 14 theme S-layer 729:735 arg1 proteins 737:744 the S-layer proteins 725:744 the S-layer proteins 725:744 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 1 15 theme material 435:442 arg1 sciences 444:451 (bio)material sciences 430:451 (bio)material sciences 430:451 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 5 16 theme hydration 1192:1200 arg1 layer 1202:1206 the adjoining hydration layer 1178:1206 the adjoining hydration layer 1178:1206 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 6 17 theme most 1309:1312 arg1 ones 1322:1325 the most general ones 1305:1325 the most general ones 1305:1325 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 1 18 theme challenging 357:367 arg1 task 369:372 nowadays a highly challenging task 339:372 nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences 339:451 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 4 19 with rSgsE 1003:1007 arg1 region 1028:1033 the preferred region 1014:1033 the preferred region being the extending glycan residues 1014:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 2 20 gly non-glycosylated 633:648 arg1 rSgsE 656:660 rSgsE 656:660 rSgsE 656:660 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 2 20 gly non-glycosylated 633:648 arg1 form 650:653 its recombinantly produced non-glycosylated form 606:653 its recombinantly produced non-glycosylated form 606:653 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 4 21 theme water 933:937 arg1 water 933:937 water 933:937 water 933:937 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 4 21 theme water 933:937 arg1 amount 923:928 almost twice the amount 906:928 almost twice the amount of water 906:937 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 2 22 theme non-glycosylated 633:648 arg1 rSgsE 656:660 rSgsE 656:660 rSgsE 656:660 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 2 22 theme non-glycosylated 633:648 arg1 form 650:653 its recombinantly produced non-glycosylated form 606:653 its recombinantly produced non-glycosylated form 606:653 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 5 23 from roughness 1227:1235 arg1 terms 1109:1113 terms 1109:1113 terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1109:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 1 24 theme surface 188:194 arg1 roughness 196:204 surface roughness 188:204 surface roughness in the nanometer scale, surface hydration and accompanied antifouling property 188:283 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 24 theme surface 188:194 arg1 etc. 286:289 etc. 286:289 etc. 286:289 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 25 from task 369:372 arg1 fields 377:382 fields 377:382 fields like microbiology, biomimetic engineering and (bio)material sciences 377:451 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 2 26 theme produced 624:631 arg1 rSgsE 656:660 rSgsE 656:660 rSgsE 656:660 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 2 26 theme produced 624:631 arg1 form 650:653 its recombinantly produced non-glycosylated form 606:653 its recombinantly produced non-glycosylated form 606:653 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 5 27 theme residues 1143:1150 arg1 effect 1122:1127 the effect 1118:1127 the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1118:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 2 28 from form 650:653 arg1 surfaces 678:685 gold sensor surfaces 666:685 gold sensor surfaces 666:685 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 2 29 from 2004/3a 594:600 arg1 surfaces 678:685 gold sensor surfaces 666:685 gold sensor surfaces 666:685 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 6 30 dep turn 1283:1286 arg1 out 1288:1290 out 1288:1290 out 1288:1290 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 2 31 theme sensor 671:676 arg1 surfaces 678:685 gold sensor surfaces 666:685 gold sensor surfaces 666:685 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 1 32 theme building 92:99 arg1 principles 101:110 the building principles 88:110 the building principles 88:110 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 4 33 theme preferred 1018:1026 arg1 region 1028:1033 the preferred region 1014:1033 the preferred region being the extending glycan residues 1014:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 3 34 theme rigid 759:763 arg1 layer 765:769 a rigid layer 757:769 a rigid layer 757:769 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 2 35 theme glycoprotein 534:545 arg1 wtSgsE 547:552 the wild-type S-layer glycoprotein wtSgsE 512:552 the wild-type S-layer glycoprotein wtSgsE 512:552 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 2 36 theme gold 666:669 arg1 surfaces 678:685 gold sensor surfaces 666:685 gold sensor surfaces 666:685 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 5 37 from behavior 1249:1256 arg1 terms 1109:1113 terms 1109:1113 terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1109:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 38 from layer 1202:1206 arg1 terms 1109:1113 terms 1109:1113 terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1109:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 0 39 theme glycosylation 13:25 arg1 Relevance 0:8 Relevance 0:8 Relevance of glycosylation of S-layer proteins for cell surface properties. 0:74 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 1 40 theme surface 295:301 arg1 structures 303:312 surface structures 295:312 surface structures 295:312 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 2 41 theme S-layer 526:532 arg1 wtSgsE 547:552 the wild-type S-layer glycoprotein wtSgsE 512:552 the wild-type S-layer glycoprotein wtSgsE 512:552 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 1 42 theme nanometer 213:221 arg1 scale 223:227 the nanometer scale 209:227 the nanometer scale 209:227 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 0 43 theme proteins 38:45 arg1 glycosylation 13:25 glycosylation 13:25 glycosylation of S-layer proteins 13:45 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 1 44 theme structures 303:312 arg1 processes 171:179 certain water-associated processes 146:179 certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures 146:312 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 2 45 theme wild-type 516:524 arg1 wtSgsE 547:552 the wild-type S-layer glycoprotein wtSgsE 512:552 the wild-type S-layer glycoprotein wtSgsE 512:552 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 1 46 dep roughness 196:204 arg1 e.g. 182:185 e.g. 182:185 e.g. 182:185 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 0 47 theme S-layer 30:36 arg1 proteins 38:45 S-layer proteins 30:45 S-layer proteins 30:45 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 5 48 theme effect 1122:1127 arg1 terms 1109:1113 terms 1109:1113 terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1109:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 49 theme glycan 1136:1141 arg1 residues 1143:1150 the glycan residues 1132:1150 the glycan residues 1132:1150 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 0 50 theme cell 51:54 arg1 properties 64:73 cell surface properties 51:73 cell surface properties 51:73 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 3 51 theme sensor 778:783 arg1 surface 785:791 the sensor surface 774:791 the sensor surface 774:791 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 0 52 gly glycosylation 13:25 arg1 proteins 38:45 S-layer proteins 30:45 S-layer proteins 30:45 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 4 53 theme wtSgsE 891:896 arg1 lattice 898:904 the wtSgsE lattice 887:904 the wtSgsE lattice 887:904 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 1 54 theme intrinsic 116:124 arg1 features 126:133 intrinsic features 116:133 intrinsic features 116:133 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 5 55 from terms 1109:1113 arg1 behavior 1249:1256 fluidic behavior 1241:1256 fluidic behavior 1241:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 55 from terms 1109:1113 arg1 roughness 1227:1235 the nanoscale roughness 1213:1235 the nanoscale roughness 1213:1235 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 55 from terms 1109:1113 arg1 layer 1202:1206 the adjoining hydration layer 1178:1206 the adjoining hydration layer 1178:1206 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 5 55 from terms 1109:1113 arg1 recrystallization 1159:1175 the recrystallization 1155:1175 the recrystallization 1155:1175 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 1 56 theme micro 320:324 arg1 organisms 326:334 (micro)organisms 319:334 (micro)organisms 319:334 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 2 57 theme Geobacillus 559:569 arg1 stearothermophilus 571:588 Geobacillus stearothermophilus 559:588 Geobacillus stearothermophilus 559:588 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 1 58 theme biomimetic 403:412 arg1 engineering 414:424 biomimetic engineering 403:424 biomimetic engineering 403:424 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 5 59 from recrystallization 1159:1175 arg1 terms 1109:1113 terms 1109:1113 terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior 1109:1256 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 4 60 with comparison 966:975 arg1 rSgsE 1003:1007 the non-glycosylated rSgsE 982:1007 the non-glycosylated rSgsE with the preferred region being the extending glycan residues 982:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 3 61 theme proteins 737:744 arg1 lattice 714:720 the proteinaceous lattice 696:720 the proteinaceous lattice of the S-layer proteins 696:744 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 5 62 theme adjoining 1182:1190 arg1 layer 1202:1206 the adjoining hydration layer 1178:1206 the adjoining hydration layer 1178:1206 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 4 63 theme glycan 1055:1060 arg1 residues 1062:1069 the extending glycan residues 1041:1069 the extending glycan residues 1041:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 1 64 theme surface 230:236 arg1 hydration 238:246 surface hydration 230:246 surface hydration 230:246 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 2 65 theme wtSgsE 547:552 arg1 recrystallization 491:507 the recrystallization 487:507 the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus 487:588 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 6 66 theme archaeal 1347:1354 arg1 lattices 1364:1371 bacterial and archaeal S-layer lattices 1333:1371 bacterial and archaeal S-layer lattices 1333:1371 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 1 67 from roughness 196:204 arg1 property 276:283 accompanied antifouling property 252:283 accompanied antifouling property 252:283 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 67 from roughness 196:204 arg1 scale 223:227 the nanometer scale 209:227 the nanometer scale 209:227 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 67 from roughness 196:204 arg1 hydration 238:246 surface hydration 230:246 surface hydration 230:246 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 1 68 theme certain 146:152 arg1 processes 171:179 certain water-associated processes 146:179 certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures 146:312 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 3 69 theme overall 830:836 arg1 film 863:866 an overall soft, highly dissipative film 827:866 an overall soft, highly dissipative film 827:866 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 3 70 theme glycan 798:803 arg1 chains 805:810 the glycan chains 794:810 the glycan chains 794:810 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 2 71 theme first 476:480 arg1 time 482:485 the first time 472:485 the first time 472:485 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 1 72 theme water-associated 154:169 arg1 processes 171:179 certain water-associated processes 146:179 certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures 146:312 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 25818946 3 73 theme soft 838:841 arg1 film 863:866 an overall soft, highly dissipative film 827:866 an overall soft, highly dissipative film 827:866 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 0 74 theme surface 56:62 arg1 properties 64:73 cell surface properties 51:73 cell surface properties 51:73 Relevance of glycosylation of S-layer proteins for cell surface properties. 25818946 3 75 theme proteinaceous 700:712 arg1 lattice 714:720 the proteinaceous lattice 696:720 the proteinaceous lattice of the S-layer proteins 696:744 Whereas the proteinaceous lattice of the S-layer proteins is forming a rigid layer on the sensor surface, the glycan chains are developing an overall soft, highly dissipative film. 25818946 4 76 gly non-glycosylated 986:1001 arg1 rSgsE 1003:1007 the non-glycosylated rSgsE 982:1007 the non-glycosylated rSgsE with the preferred region being the extending glycan residues 982:1069 Interestingly, to the wtSgsE lattice almost twice the amount of water is bound and/or coupled in comparison with the non-glycosylated rSgsE with the preferred region being the extending glycan residues. 25818946 6 77 theme S-layer 1356:1362 arg1 lattices 1364:1371 bacterial and archaeal S-layer lattices 1333:1371 bacterial and archaeal S-layer lattices 1333:1371 The latter features may turn out to be one of the most general ones among bacterial and archaeal S-layer lattices. 25818946 2 78 gly glycoprotein 534:545 arg1 glycoprotein 534:545 the wild-type S-layer glycoprotein wtSgsE 512:552 the wild-type S-layer glycoprotein wtSgsE 512:552 Here, we show for the first time the recrystallization of the wild-type S-layer glycoprotein wtSgsE from Geobacillus stearothermophilus NRS 2004/3a and its recombinantly produced non-glycosylated form, rSgsE, on gold sensor surfaces. 25818946 5 79 theme nanoscale 1217:1225 arg1 roughness 1227:1235 the nanoscale roughness 1213:1235 the nanoscale roughness 1213:1235 The present results are discussed in terms of the effect of the glycan residues on the recrystallization, the adjoining hydration layer, and the nanoscale roughness and fluidic behavior. 25818946 1 80 theme accompanied 252:262 arg1 property 276:283 accompanied antifouling property 252:283 accompanied antifouling property 252:283 Elucidating the building principles and intrinsic features modulating certain water-associated processes (e.g., surface roughness in the nanometer scale, surface hydration and accompanied antifouling property, etc.) of surface structures from (micro)organisms is nowadays a highly challenging task in fields like microbiology, biomimetic engineering and (bio)material sciences. 28434712 1 0 theme unknown 153:159 arg1 by-product 173:182 a previously unknown imidazoline by-product 140:182 a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1 140:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 1 1 theme N-linked 275:282 arg1 glycans 284:290 enzymatically released N-linked glycans 252:290 enzymatically released N-linked glycans of a human IgG1 252:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 6 2 theme fragment 1332:1339 arg1 ions 1341:1344 high resolution mass fragment ions 1311:1344 high resolution mass fragment ions 1311:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 1 3 theme imidazoline 161:171 arg1 by-product 173:182 a previously unknown imidazoline by-product 140:182 a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1 140:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 6 4 theme reducing 1268:1275 arg1 end 1277:1279 the reducing end 1264:1279 the reducing end of the glycan as confirmed by high resolution mass fragment ions 1264:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 7 5 theme reductant 1386:1394 arg1 concentration 1396:1408 the reductant concentration 1382:1408 the reductant concentration 1382:1408 Imidazoline content decreased when the reductant concentration was increased. 28434712 6 6 theme mass 1327:1330 arg1 ions 1341:1344 high resolution mass fragment ions 1311:1344 high resolution mass fragment ions 1311:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 5 7 theme molecular 1006:1014 arg1 location 1016:1023 the molecular location 1002:1023 the molecular location of loss of water 1002:1040 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 5 7 theme molecular 1006:1014 arg1 straightforward 1050:1064 straightforward 1050:1064 straightforward 1050:1064 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 3 8 theme -labeled 603:610 arg1 well-resolved 625:637 well-resolved 625:637 well-resolved 625:637 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 8 theme -labeled 603:610 arg1 glycans 612:618 the 2-aminobenzoic acid (2-AA)-labeled glycans 573:618 the 2-aminobenzoic acid (2-AA)-labeled glycans 573:618 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 6 9 theme glycan 1288:1293 arg1 end 1277:1279 the reducing end 1264:1279 the reducing end of the glycan as confirmed by high resolution mass fragment ions 1264:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 6 10 theme resolution 1316:1325 arg1 mass 1327:1330 high resolution mass 1311:1330 high resolution mass fragment ions 1311:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 1 11 link N-linked 275:282 arg1 glycans 284:290 enzymatically released N-linked glycans 252:290 enzymatically released N-linked glycans of a human IgG1 252:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 6 12 theme ring 1238:1241 arg1 impurity 1211:1218 the impurity 1207:1218 the impurity 1207:1218 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 6 12 theme ring 1238:1241 arg1 structure 1243:1251 an imidazoline ring structure 1223:1251 an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions 1223:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 6 13 theme standard 1162:1169 arg1 A2G0F 1156:1160 A2G0F standard 1156:1169 A2G0F standard 1156:1169 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 0 14 from glycans 44:50 arg1 by-product 28:37 an imidazoline by-product 13:37 an imidazoline by-product from glycans 13:50 Evidence for an imidazoline by-product from glycans using tandem mass spectrometry. 28434712 6 15 theme high 1311:1314 arg1 mass 1327:1330 high resolution mass 1311:1330 high resolution mass fragment ions 1311:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 1 16 dep separation 107:116 arg1 the 103:105 the 103:105 the 103:105 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 6 17 theme Model 1119:1123 arg1 studies 1125:1131 Model studies 1119:1131 Model studies 1119:1131 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 1 18 theme by-product 173:182 arg1 identification 122:135 identification 122:135 identification 122:135 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 1 18 theme by-product 173:182 arg1 separation 107:116 separation 107:116 separation 107:116 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 5 19 theme loss 1028:1031 arg1 location 1016:1023 the molecular location 1002:1023 the molecular location of loss of water 1002:1040 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 5 19 theme loss 1028:1031 arg1 straightforward 1050:1064 straightforward 1050:1064 straightforward 1050:1064 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 8 20 theme imidazoline 1455:1465 arg1 structure 1467:1475 the imidazoline structure 1451:1475 the imidazoline structure 1451:1475 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 2 21 theme 2-picoline 427:436 arg1 borane 438:443 2-picoline borane 427:443 2-picoline borane 427:443 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 5 22 theme water 986:990 arg1 molecule 992:999 a water molecule 984:999 a water molecule 984:999 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 8 23 theme chemical 1583:1590 arg1 susceptibility 1592:1605 chemical susceptibility 1583:1605 chemical susceptibility 1583:1605 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 3 24 theme liquid 470:475 arg1 UPLC 493:496 UPLC 493:496 UPLC 493:496 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 24 theme liquid 470:475 arg1 chromatography 477:490 ultra performance liquid chromatography 452:490 ultra performance liquid chromatography (UPLC) 452:497 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 9 25 link N-linked 1746:1753 arg1 glycans 1755:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 This study is the first to identify these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics. 28434712 1 26 theme human 297:301 arg1 IgG1 303:306 a human IgG1 295:306 a human IgG1 295:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 4 27 theme minor 808:812 arg1 species 814:820 this minor species 803:820 this minor species (∼2%) 803:826 It was found that this minor species (∼2%) was 18.0105u less than the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak, abbreviated as A2G0F at 1582.5899u. 28434712 4 27 theme minor 808:812 arg1 % 825:825 ∼2% 823:825 ∼2% 823:825 It was found that this minor species (∼2%) was 18.0105u less than the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak, abbreviated as A2G0F at 1582.5899u. 28434712 5 28 from consideration 1069:1081 arg1 location 1016:1023 the molecular location 1002:1023 the molecular location of loss of water 1002:1040 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 5 28 from consideration 1069:1081 arg1 straightforward 1050:1064 straightforward 1050:1064 straightforward 1050:1064 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 9 29 theme N-acetylglucosamine-type 1721:1744 arg1 glycans 1755:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 This study is the first to identify these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics. 28434712 0 30 theme imidazoline 16:26 arg1 by-product 28:37 an imidazoline by-product 13:37 an imidazoline by-product from glycans 13:50 Evidence for an imidazoline by-product from glycans using tandem mass spectrometry. 28434712 1 31 theme IgG1 303:306 arg1 glycans 284:290 enzymatically released N-linked glycans 252:290 enzymatically released N-linked glycans of a human IgG1 252:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 8 32 theme accuracy 1524:1531 arg1 HRAM 1552:1555 HRAM 1552:1555 HRAM 1552:1555 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 8 32 theme accuracy 1524:1531 arg1 spectrometry 1538:1549 high accuracy mass spectrometry 1519:1549 high accuracy mass spectrometry (HRAM) 1519:1556 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 9 33 theme minor 1683:1687 arg1 species 1689:1695 these minor species 1677:1695 these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics 1677:1776 This study is the first to identify these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics. 28434712 4 34 theme GlcNAc2Man3GlcNAc2Fuc 872:892 arg1 peak 894:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 It was found that this minor species (∼2%) was 18.0105u less than the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak, abbreviated as A2G0F at 1582.5899u. 28434712 1 35 theme fluorescent 205:215 arg1 labeling 217:224 fluorescent labeling 205:224 the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1 201:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 5 36 theme molecule 992:999 arg1 mass 976:979 the mass 972:979 the mass of a water molecule 972:999 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 3 37 theme 2-aminobenzoic 577:590 arg1 acid 592:595 2-aminobenzoic acid 577:595 2-aminobenzoic acid 577:595 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 38 theme ionization 727:736 arg1 ESI-MS 757:762 ESI-MS 757:762 ESI-MS 757:762 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 38 theme ionization 727:736 arg1 spectrometry 743:754 electrospray ionization mass spectrometry 714:754 electrospray ionization mass spectrometry (ESI-MS) 714:763 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 2 39 theme imidazoline 313:323 arg1 by-product 325:334 The imidazoline by-product 309:334 The imidazoline by-product 309:334 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 5 40 from straightforward 1050:1064 arg1 consideration 1069:1081 consideration 1069:1081 consideration of the biantennary A2G0F structure 1069:1116 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 1 41 theme labeling 217:224 arg1 procedure 226:234 the fluorescent labeling procedure 201:234 the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1 201:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 28434712 3 42 theme ultra 452:456 arg1 UPLC 493:496 UPLC 493:496 UPLC 493:496 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 42 theme ultra 452:456 arg1 chromatography 477:490 ultra performance liquid chromatography 452:490 ultra performance liquid chromatography (UPLC) 452:497 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 9 43 from biologics 1768:1776 arg1 impact 1710:1715 impact 1710:1715 impact 1710:1715 This study is the first to identify these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics. 28434712 3 44 with conjunction 502:512 arg1 chromatography 549:562 hydrophilic interaction-based chromatography 519:562 hydrophilic interaction-based chromatography (HILIC) 519:570 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 44 with conjunction 502:512 arg1 HILIC 565:569 HILIC 565:569 HILIC 565:569 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 45 theme mass 738:741 arg1 ESI-MS 757:762 ESI-MS 757:762 ESI-MS 757:762 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 45 theme mass 738:741 arg1 spectrometry 743:754 electrospray ionization mass spectrometry 714:754 electrospray ionization mass spectrometry (ESI-MS) 714:763 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 46 theme imidazoline 644:654 arg1 by-products 656:666 imidazoline by-products 644:666 imidazoline by-products 644:666 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 47 from by-products 656:666 arg1 well-resolved 625:637 well-resolved 625:637 well-resolved 625:637 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 47 from by-products 656:666 arg1 glycans 612:618 the 2-aminobenzoic acid (2-AA)-labeled glycans 573:618 the 2-aminobenzoic acid (2-AA)-labeled glycans 573:618 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 48 theme performance 458:468 arg1 UPLC 493:496 UPLC 493:496 UPLC 493:496 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 48 theme performance 458:468 arg1 chromatography 477:490 ultra performance liquid chromatography 452:490 ultra performance liquid chromatography (UPLC) 452:497 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 8 49 theme mass 1533:1536 arg1 HRAM 1552:1555 HRAM 1552:1555 HRAM 1552:1555 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 8 49 theme mass 1533:1536 arg1 spectrometry 1538:1549 high accuracy mass spectrometry 1519:1549 high accuracy mass spectrometry (HRAM) 1519:1556 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 4 50 theme peak 867:870 arg1 peak 894:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 It was found that this minor species (∼2%) was 18.0105u less than the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak, abbreviated as A2G0F at 1582.5899u. 28434712 8 51 theme high 1519:1522 arg1 HRAM 1552:1555 HRAM 1552:1555 HRAM 1552:1555 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 8 51 theme high 1519:1522 arg1 spectrometry 1538:1549 high accuracy mass spectrometry 1519:1549 high accuracy mass spectrometry (HRAM) 1519:1556 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 7 52 theme Imidazoline 1347:1357 arg1 content 1359:1365 Imidazoline content 1347:1365 Imidazoline content 1347:1365 Imidazoline content decreased when the reductant concentration was increased. 28434712 5 53 theme mass 947:950 arg1 loss 952:955 this mass loss 942:955 this mass loss 942:955 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 3 54 theme electrospray 714:725 arg1 ESI-MS 757:762 ESI-MS 757:762 ESI-MS 757:762 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 54 theme electrospray 714:725 arg1 spectrometry 743:754 electrospray ionization mass spectrometry 714:754 electrospray ionization mass spectrometry (ESI-MS) 714:763 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 2 55 theme amination 368:376 arg1 procedure 378:386 the reductive amination procedure 354:386 the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane 354:443 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 5 56 theme biantennary 1090:1100 arg1 structure 1108:1116 the biantennary A2G0F structure 1086:1116 the biantennary A2G0F structure 1086:1116 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 2 57 theme reductive 358:366 arg1 amination 368:376 the reductive amination 354:376 the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane 354:443 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 8 58 theme high 1502:1505 arg1 resolution 1507:1516 high resolution 1502:1516 high resolution 1502:1516 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 2 59 with procedure 378:386 arg1 cyanoborohydride 407:422 sodium cyanoborohydride 400:422 sodium cyanoborohydride 400:422 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 2 59 with procedure 378:386 arg1 borane 438:443 2-picoline borane 427:443 2-picoline borane 427:443 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 3 60 theme direct 682:687 arg1 identification 689:702 direct identification 682:702 direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation 682:782 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 61 theme hydrophilic 519:529 arg1 chromatography 549:562 hydrophilic interaction-based chromatography 519:562 hydrophilic interaction-based chromatography (HILIC) 519:570 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 61 theme hydrophilic 519:529 arg1 HILIC 565:569 HILIC 565:569 HILIC 565:569 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 6 62 from end 1277:1279 arg1 located 1253:1259 located 1253:1259 located 1253:1259 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 5 63 theme A2G0F 1102:1106 arg1 structure 1108:1116 the biantennary A2G0F structure 1086:1116 the biantennary A2G0F structure 1086:1116 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 5 64 theme water 1036:1040 arg1 loss 1028:1031 loss 1028:1031 loss of water 1028:1040 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 3 65 theme interaction-based 531:547 arg1 chromatography 549:562 hydrophilic interaction-based chromatography 519:562 hydrophilic interaction-based chromatography (HILIC) 519:570 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 3 65 theme interaction-based 531:547 arg1 HILIC 565:569 HILIC 565:569 HILIC 565:569 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 9 66 theme N-linked 1746:1753 arg1 glycans 1755:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 all N-acetylglucosamine-type N-linked glycans 1717:1761 This study is the first to identify these minor species which likely impact all N-acetylglucosamine-type N-linked glycans from biologics. 28434712 0 67 theme mass 65:68 arg1 spectrometry 70:81 tandem mass spectrometry 58:81 tandem mass spectrometry 58:81 Evidence for an imidazoline by-product from glycans using tandem mass spectrometry. 28434712 5 68 theme structure 1108:1116 arg1 consideration 1069:1081 consideration 1069:1081 consideration of the biantennary A2G0F structure 1069:1116 While this mass loss corresponds to the mass of a water molecule, the molecular location of loss of water was not straightforward in consideration of the biantennary A2G0F structure. 28434712 4 69 theme neighboring 855:865 arg1 peak 894:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak 851:897 It was found that this minor species (∼2%) was 18.0105u less than the neighboring peak GlcNAc2Man3GlcNAc2Fuc peak, abbreviated as A2G0F at 1582.5899u. 28434712 8 70 theme labeled 1624:1630 arg1 tracers 1632:1638 isotopically labeled tracers 1611:1638 isotopically labeled tracers 1611:1638 To conclude, evidence for the imidazoline structure was accomplished through high resolution, high accuracy mass spectrometry (HRAM), and experiments showing chemical susceptibility and isotopically labeled tracers. 28434712 2 71 theme sodium 400:405 arg1 cyanoborohydride 407:422 sodium cyanoborohydride 400:422 sodium cyanoborohydride 400:422 The imidazoline by-product was generated via the reductive amination procedure with either sodium cyanoborohydride or 2-picoline borane. 28434712 6 72 theme imidazoline 1226:1236 arg1 impurity 1211:1218 the impurity 1207:1218 the impurity 1207:1218 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 6 72 theme imidazoline 1226:1236 arg1 structure 1243:1251 an imidazoline ring structure 1223:1251 an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions 1223:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 0 73 theme tandem 58:63 arg1 spectrometry 70:81 tandem mass spectrometry 58:81 tandem mass spectrometry 58:81 Evidence for an imidazoline by-product from glycans using tandem mass spectrometry. 28434712 6 74 theme located 1253:1259 arg1 impurity 1211:1218 the impurity 1207:1218 the impurity 1207:1218 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 6 74 theme located 1253:1259 arg1 structure 1243:1251 an imidazoline ring structure 1223:1251 an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions 1223:1344 Model studies were carried out using A2G0F standard and N-acetyllactosamine to identify the impurity as an imidazoline ring structure located at the reducing end of the glycan as confirmed by high resolution mass fragment ions. 28434712 3 75 dep -labeled 603:610 arg1 acid 592:595 2-aminobenzoic acid 577:595 2-aminobenzoic acid 577:595 Using ultra performance liquid chromatography (UPLC) in conjunction with hydrophilic interaction-based chromatography (HILIC), the 2-aminobenzoic acid (2-AA)-labeled glycans were well-resolved from imidazoline by-products to facilitate direct identification utilizing electrospray ionization mass spectrometry (ESI-MS) with fragmentation. 28434712 1 76 theme released 266:273 arg1 glycans 284:290 enzymatically released N-linked glycans 252:290 enzymatically released N-linked glycans of a human IgG1 252:306 Herein is reported the separation and identification of a previously unknown imidazoline by-product originating from the fluorescent labeling procedure when applied to enzymatically released N-linked glycans of a human IgG1. 26457517 2 0 theme Clostridium 358:368 arg1 cellulovorans 370:382 Clostridium cellulovorans 358:382 Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules 358:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 4 1 from subsites 744:751 arg1 structure 769:777 the structure 765:777 the structure of the ExgS-triethylene glycol complex shown here 765:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 4 1 from subsites 744:751 arg1 located 721:727 located 721:727 located 721:727 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 1 2 theme reducing 185:192 arg1 end 194:196 the reducing end 181:196 the reducing end of cellulose 181:209 Exoglucanase/cellobiohydrolase (EC 3.2.1.176) hydrolyzes a β-1,4-glycosidic bond from the reducing end of cellulose and releases cellobiose as the major product. 26457517 5 3 from Modelling 830:838 arg1 site 881:884 the active site 870:884 the active site of the ExgS-cellobiose structure 870:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 3 from Modelling 830:838 arg1 structure 909:917 the ExgS-cellobiose structure 889:917 the ExgS-cellobiose structure 889:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 3 4 theme complex 570:576 arg1 structure 578:586 the enzyme-cellotetraose complex structure 545:586 the enzyme-cellotetraose complex structure 545:586 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 5 5 theme nucleophilic 981:992 arg1 donor 956:960 a proton donor 947:960 a proton donor 947:960 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 5 theme nucleophilic 981:992 arg1 Glu50 933:937 Glu50 933:937 Glu50 933:937 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 5 theme nucleophilic 981:992 arg1 role 994:997 a nucleophilic role 979:997 a nucleophilic role 979:997 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 3 6 theme structure 578:586 arg1 cleft 536:540 the open active-site cleft 515:540 the open active-site cleft of the enzyme-cellotetraose complex structure 515:586 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 2 7 theme hydrolase 306:314 arg1 ExgS 347:350 ExgS 347:350 ExgS 347:350 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 2 7 theme hydrolase 306:314 arg1 S 344:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S 293:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) 293:351 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 1 8 theme β-1,4-glycosidic 154:169 arg1 bond 171:174 a β-1,4-glycosidic bond 152:174 a β-1,4-glycosidic bond from the reducing end of cellulose 152:209 Exoglucanase/cellobiohydrolase (EC 3.2.1.176) hydrolyzes a β-1,4-glycosidic bond from the reducing end of cellulose and releases cellobiose as the major product. 26457517 5 9 theme active 874:879 arg1 site 881:884 the active site 870:884 the active site of the ExgS-cellobiose structure 870:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 9 theme active 874:879 arg1 structure 909:917 the ExgS-cellobiose structure 889:917 the ExgS-cellobiose structure 889:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 2 10 theme glycosyl 297:304 arg1 ExgS 347:350 ExgS 347:350 ExgS 347:350 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 2 10 theme glycosyl 297:304 arg1 S 344:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S 293:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) 293:351 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 5 11 theme structure 909:917 arg1 site 881:884 the active site 870:884 the active site of the ExgS-cellobiose structure 870:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 11 theme structure 909:917 arg1 structure 909:917 the ExgS-cellobiose structure 889:917 the ExgS-cellobiose structure 889:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 0 12 from cellulovorans 44:56 arg1 cleavage 85:92 cleavage 85:92 cleavage 85:92 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 12 from cellulovorans 44:56 arg1 Structures 0:9 Structures 0:9 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 0:93 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 12 from cellulovorans 44:56 arg1 binding 73:79 cellotetraose binding 59:79 cellotetraose binding 59:79 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 3 13 theme enzyme-cellotetraose 549:568 arg1 structure 578:586 the enzyme-cellotetraose complex structure 545:586 the enzyme-cellotetraose complex structure 545:586 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 2 14 theme cellotetraose 401:413 arg1 molecules 438:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 4 15 theme glycol 701:706 arg1 molecule 708:715 one pentaethylene glycol molecule 683:715 one pentaethylene glycol molecule 683:715 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 2 16 theme cellobiohydrolase 326:342 arg1 ExgS 347:350 ExgS 347:350 ExgS 347:350 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 2 16 theme cellobiohydrolase 326:342 arg1 S 344:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S 293:344 the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) 293:351 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 1 17 theme cellulose 201:209 arg1 end 194:196 the reducing end 181:196 the reducing end of cellulose 181:209 Exoglucanase/cellobiohydrolase (EC 3.2.1.176) hydrolyzes a β-1,4-glycosidic bond from the reducing end of cellulose and releases cellobiose as the major product. 26457517 3 18 theme enzymatic 603:611 arg1 function 624:631 an enzymatic hydrolysis function 600:631 an enzymatic hydrolysis function 600:631 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 2 19 theme cellobiose 389:398 arg1 molecules 438:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 3 20 dep subsites 493:500 arg1 +2 509:510 +2 509:510 +2 509:510 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 3 20 dep subsites 493:500 arg1 +1 502:503 +1 502:503 +1 502:503 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 2 21 from cellulovorans 370:382 arg1 structures 279:288 Three complex crystal structures 257:288 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules 257:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 4 22 theme ExgS-triethylene 786:801 arg1 complex 810:816 the ExgS-triethylene glycol complex 782:816 the ExgS-triethylene glycol complex shown here 782:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 0 23 theme exoglucanase 14:25 arg1 cleavage 85:92 cleavage 85:92 cleavage 85:92 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 23 theme exoglucanase 14:25 arg1 Structures 0:9 Structures 0:9 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 0:93 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 23 theme exoglucanase 14:25 arg1 binding 73:79 cellotetraose binding 59:79 cellotetraose binding 59:79 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 3 24 theme active-site 524:534 arg1 cleft 536:540 the open active-site cleft 515:540 the open active-site cleft of the enzyme-cellotetraose complex structure 515:586 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 5 25 theme ExgS-cellobiose 893:907 arg1 structure 909:917 the ExgS-cellobiose structure 889:917 the ExgS-cellobiose structure 889:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 3 26 theme open 519:522 arg1 cleft 536:540 the open active-site cleft 515:540 the open active-site cleft of the enzyme-cellotetraose complex structure 515:586 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 4 27 theme pentaethylene 687:699 arg1 molecule 708:715 one pentaethylene glycol molecule 683:715 one pentaethylene glycol molecule 683:715 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 2 28 theme glycol 431:436 arg1 molecules 438:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 0 29 theme Clostridium 32:42 arg1 cellulovorans 44:56 Clostridium cellulovorans 32:56 Clostridium cellulovorans 32:56 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 4 30 theme glycol 803:808 arg1 complex 810:816 the ExgS-triethylene glycol complex 782:816 the ExgS-triethylene glycol complex shown here 782:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 2 31 theme triethylene 419:429 arg1 molecules 438:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 0 32 theme cellotetraose 59:71 arg1 binding 73:79 cellotetraose binding 59:79 cellotetraose binding 59:79 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 32 theme cellotetraose 59:71 arg1 Structures 0:9 Structures 0:9 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 0:93 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 4 33 located located 721:727 arg1 subsites 744:751 active-site subsites -2 to -6 732:760 active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here 732:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 4 33 located located 721:727 arg2 molecules 669:677 three triethylene glycol molecules 644:677 three triethylene glycol molecules 644:677 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 4 33 located located 721:727 arg2 molecule 708:715 one pentaethylene glycol molecule 683:715 one pentaethylene glycol molecule 683:715 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 3 34 theme hydrolysis 613:622 arg1 function 624:631 an enzymatic hydrolysis function 600:631 an enzymatic hydrolysis function 600:631 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 5 35 theme glucose 843:849 arg1 Modelling 830:838 Modelling 830:838 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure 830:917 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 36 theme proton 949:954 arg1 donor 956:960 a proton donor 947:960 a proton donor 947:960 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 36 theme proton 949:954 arg1 Glu50 933:937 Glu50 933:937 Glu50 933:937 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 5 36 theme proton 949:954 arg1 role 994:997 a nucleophilic role 979:997 a nucleophilic role 979:997 Modelling of glucose into subsite -1 in the active site of the ExgS-cellobiose structure revealed that Glu50 acts as a proton donor and Asp222 plays a nucleophilic role. 26457517 0 37 dep Structures 0:9 arg1 cleavage 85:92 cleavage 85:92 cleavage 85:92 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 37 dep Structures 0:9 arg1 binding 73:79 cellotetraose binding 59:79 cellotetraose binding 59:79 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 0 37 dep Structures 0:9 arg1 Structures 0:9 Structures 0:9 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 0:93 Structures of exoglucanase from Clostridium cellulovorans: cellotetraose binding and cleavage. 26457517 4 38 dep -6 759:760 arg1 to 756:757 to 756:757 to 756:757 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 1 39 theme major 242:246 arg1 product 248:254 the major product 238:254 the major product 238:254 Exoglucanase/cellobiohydrolase (EC 3.2.1.176) hydrolyzes a β-1,4-glycosidic bond from the reducing end of cellulose and releases cellobiose as the major product. 26457517 4 40 theme complex 810:816 arg1 structure 769:777 the structure 765:777 the structure of the ExgS-triethylene glycol complex shown here 765:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 2 41 theme S 344:344 arg1 structures 279:288 Three complex crystal structures 257:288 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules 257:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 2 42 with cellulovorans 370:382 arg1 molecules 438:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 cellobiose, cellotetraose and triethylene glycol molecules 389:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 2 43 theme crystal 271:277 arg1 structures 279:288 Three complex crystal structures 257:288 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules 257:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 4 44 theme active-site 732:742 arg1 subsites 744:751 active-site subsites -2 to -6 732:760 active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here 732:827 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 3 45 theme product 465:471 arg1 cellobiose 473:482 The product cellobiose 461:482 The product cellobiose 461:482 The product cellobiose occupies subsites +1 and +2 in the open active-site cleft of the enzyme-cellotetraose complex structure, indicating an enzymatic hydrolysis function. 26457517 4 46 theme glycol 662:667 arg1 molecules 669:677 three triethylene glycol molecules 644:677 three triethylene glycol molecules 644:677 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 26457517 2 47 theme complex 263:269 arg1 structures 279:288 Three complex crystal structures 257:288 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules 257:446 Three complex crystal structures of the glycosyl hydrolase 48 (GH48) cellobiohydrolase S (ExgS) from Clostridium cellulovorans with cellobiose, cellotetraose and triethylene glycol molecules were solved. 26457517 1 48 from end 194:196 arg1 bond 171:174 a β-1,4-glycosidic bond 152:174 a β-1,4-glycosidic bond from the reducing end of cellulose 152:209 Exoglucanase/cellobiohydrolase (EC 3.2.1.176) hydrolyzes a β-1,4-glycosidic bond from the reducing end of cellulose and releases cellobiose as the major product. 26457517 4 49 theme triethylene 650:660 arg1 molecules 669:677 three triethylene glycol molecules 644:677 three triethylene glycol molecules 644:677 Moreover, three triethylene glycol molecules and one pentaethylene glycol molecule are located at active-site subsites -2 to -6 in the structure of the ExgS-triethylene glycol complex shown here. 29019687 0 0 theme Strong 81:86 arg1 Coupling 96:103 Strong Dynamic Coupling 81:103 Strong Dynamic Coupling between Glycans and Protein Core 81:136 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 5 1 from glycans 787:793 arg1 simulations 802:812 our simulations 798:812 our simulations 798:812 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 5 2 from locations 886:894 arg1 dynamics 853:860 the dynamics 849:860 the dynamics of glycans at different locations on NMDARs 849:904 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 5 2 from locations 886:894 arg1 NMDARs 899:904 NMDARs 899:904 NMDARs 899:904 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 5 2 from locations 886:894 arg1 different 923:931 different 923:931 different 923:931 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 9 3 theme receptors 1635:1643 arg1 structure 1599:1607 the structure 1595:1607 the structure 1595:1607 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 9 3 theme receptors 1635:1643 arg1 dynamics 1617:1624 the dynamics 1613:1624 the dynamics of these receptors 1613:1643 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 0 4 from Dynamics 10:17 arg1 Domain 48:53 Ligand-Binding Domain 33:53 Ligand-Binding Domain of NMDA Receptors 33:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 0 4 from Dynamics 10:17 arg1 Receptors 63:71 NMDA Receptors 58:71 NMDA Receptors 58:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 4 5 theme aggregate 644:652 arg1 μs 659:660 aggregate 86.5 μs long 644:665 aggregate 86.5 μs long 644:665 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 4 6 theme glycosylated 675:686 arg1 LBD 688:690 the glycosylated LBD 671:690 the glycosylated LBD of the GluN1 subunit of the NMDAR 671:724 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 6 7 theme other 1095:1099 arg1 cases 1101:1105 other cases 1095:1105 other cases 1095:1105 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 7 8 from glycans 1216:1222 arg1 scales 1180:1185 time scales 1175:1185 time scales of conformational changes in glycans on the GluN1 LBD of NMDARs 1175:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 3 9 theme NMDA 486:489 arg1 NMDARs 502:507 NMDARs 502:507 NMDARs 502:507 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 9 theme NMDA 486:489 arg1 receptors 491:499 NMDA receptors 486:499 NMDA receptors (NMDARs) 486:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 4 10 theme GluN1 699:703 arg1 subunit 705:711 the GluN1 subunit 695:711 the GluN1 subunit of the NMDAR 695:724 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 4 11 theme LBD 688:690 arg1 trajectories 627:638 molecular dynamics trajectories 608:638 molecular dynamics trajectories 608:638 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 0 12 theme Dynamic 88:94 arg1 Coupling 96:103 Strong Dynamic Coupling 81:103 Strong Dynamic Coupling between Glycans and Protein Core 81:136 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 8 13 theme clamshell-like 1407:1420 arg1 opening 1422:1428 the physiologically important clamshell-like opening 1377:1428 the physiologically important clamshell-like opening 1377:1428 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 6 14 theme glycan 1151:1156 arg1 bending 1138:1144 a head-to-tail bending 1123:1144 a head-to-tail bending of a glycan 1123:1156 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 7 15 from scales 1180:1185 arg1 LBD 1237:1239 the GluN1 LBD 1227:1239 the GluN1 LBD of NMDARs 1227:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 7 15 from scales 1180:1185 arg1 glycans 1216:1222 glycans 1216:1222 glycans 1216:1222 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 4 16 from trajectories 627:638 arg1 μs 659:660 aggregate 86.5 μs long 644:665 aggregate 86.5 μs long 644:665 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 8 17 theme important 1397:1405 arg1 opening 1422:1428 the physiologically important clamshell-like opening 1377:1428 the physiologically important clamshell-like opening 1377:1428 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 3 18 theme ligand 455:460 arg1 LBD 478:480 LBD 478:480 LBD 478:480 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 18 theme ligand 455:460 arg1 domain 470:475 the ligand binding domain 451:475 the ligand binding domain (LBD) of NMDA receptors (NMDARs) 451:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 18 theme ligand 455:460 arg1 receptors 491:499 NMDA receptors 486:499 NMDA receptors (NMDARs) 486:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 5 19 theme different 876:884 arg1 locations 886:894 different locations 876:894 different locations on NMDARs 876:904 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 7 20 theme conformational 1190:1203 arg1 changes 1205:1211 conformational changes 1190:1211 conformational changes in glycans 1190:1222 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 4 21 theme molecular 608:616 arg1 trajectories 627:638 molecular dynamics trajectories 608:638 molecular dynamics trajectories 608:638 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 8 22 from changes 1339:1345 arg1 glycans 1354:1360 the glycans 1350:1360 the glycans 1350:1360 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 9 23 theme NMDARs 1541:1546 arg1 models 1531:1536 computational models 1517:1536 computational models of NMDARs 1517:1546 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 4 24 theme subunit 705:711 arg1 LBD 688:690 the glycosylated LBD 671:690 the glycosylated LBD of the GluN1 subunit of the NMDAR 671:724 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 2 25 theme glycans 376:382 arg1 structure 324:332 structure 324:332 structure 324:332 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 2 25 theme glycans 376:382 arg1 relevance 363:371 physiological relevance 349:371 physiological relevance 349:371 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 2 25 theme glycans 376:382 arg1 dynamics 335:342 dynamics 335:342 dynamics 335:342 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 6 26 theme relative 1062:1069 arg1 glycans 1054:1060 glycans 1054:1060 glycans relative to each other 1054:1083 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 3 27 theme intramolecular 523:536 arg1 glycans 440:446 certain glycans 432:446 certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) 432:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 27 theme intramolecular 523:536 arg1 potentiators 538:549 intramolecular potentiators 523:549 intramolecular potentiators 523:549 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 1 28 theme key 178:180 arg1 receptors 167:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 1 28 theme key 178:180 arg1 receptors 191:199 key neuronal receptors 178:199 key neuronal receptors playing the central role in learning and memory 178:247 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 0 29 theme Simulated 0:8 arg1 Dynamics 10:17 Simulated Dynamics 0:17 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors 0:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 7 30 theme microseconds 1298:1309 arg1 hundreds 1286:1293 hundreds 1286:1293 at least hundreds of microseconds 1277:1309 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 6 31 theme head-to-tail 1125:1136 arg1 bending 1138:1144 a head-to-tail bending 1123:1144 a head-to-tail bending of a glycan 1123:1156 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 6 32 theme glycans 1054:1060 arg1 parts 1045:1049 parts 1045:1049 parts of glycans relative to each other 1045:1083 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 1 33 theme neuronal 182:189 arg1 receptors 167:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 1 33 theme neuronal 182:189 arg1 receptors 191:199 key neuronal receptors 178:199 key neuronal receptors playing the central role in learning and memory 178:247 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 0 34 theme Glycans 22:28 arg1 Dynamics 10:17 Simulated Dynamics 0:17 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors 0:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 6 35 theme various 985:991 arg1 glycans 993:999 various glycans 985:999 various glycans 985:999 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 0 36 theme Protein 125:131 arg1 Core 133:136 Protein Core 125:136 Protein Core 125:136 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 9 37 theme integral 1488:1495 arg1 part 1497:1500 an integral part 1485:1500 an integral part of NMDARs 1485:1510 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 9 37 theme integral 1488:1495 arg1 glycans 1473:1479 glycans 1473:1479 glycans 1473:1479 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 8 38 theme GluN1 1449:1453 arg1 domain 1459:1464 the GluN1 LBD domain 1445:1464 the GluN1 LBD domain 1445:1464 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 5 39 dep same 836:839 arg1 Man5 842:845 Man5 842:845 Man5 842:845 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 2 40 dep structure 324:332 arg1 the 320:322 the 320:322 the 320:322 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 1 41 theme N-Methyl-d-aspartate 139:158 arg1 receptors 167:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 1 41 theme N-Methyl-d-aspartate 139:158 arg1 receptors 191:199 key neuronal receptors 178:199 key neuronal receptors playing the central role in learning and memory 178:247 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 0 42 theme Ligand-Binding 33:46 arg1 Domain 48:53 Ligand-Binding Domain 33:53 Ligand-Binding Domain of NMDA Receptors 33:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 0 42 theme Ligand-Binding 33:46 arg1 Receptors 63:71 NMDA Receptors 58:71 NMDA Receptors 58:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 7 43 theme time 1175:1178 arg1 scales 1180:1185 time scales 1175:1185 time scales of conformational changes in glycans on the GluN1 LBD of NMDARs 1175:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 4 44 theme dynamics 618:625 arg1 trajectories 627:638 molecular dynamics trajectories 608:638 molecular dynamics trajectories 608:638 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 8 45 theme domain 1459:1464 arg1 closing 1434:1440 closing 1434:1440 closing 1434:1440 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 8 45 theme domain 1459:1464 arg1 opening 1422:1428 the physiologically important clamshell-like opening 1377:1428 the physiologically important clamshell-like opening 1377:1428 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 8 46 theme conformational 1324:1337 arg1 changes 1339:1345 the conformational changes 1320:1345 the conformational changes in the glycans 1320:1360 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 3 47 from glycans 440:446 arg1 LBD 478:480 LBD 478:480 LBD 478:480 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 47 from glycans 440:446 arg1 domain 470:475 the ligand binding domain 451:475 the ligand binding domain (LBD) of NMDA receptors (NMDARs) 451:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 47 from glycans 440:446 arg1 receptors 491:499 NMDA receptors 486:499 NMDA receptors (NMDARs) 486:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 1 48 theme central 213:219 arg1 role 221:224 the central role 209:224 the central role 209:224 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 8 49 theme LBD 1455:1457 arg1 domain 1459:1464 the GluN1 LBD domain 1445:1464 the GluN1 LBD domain 1445:1464 Some of the conformational changes in the glycans correlate with the physiologically important clamshell-like opening and closing of the GluN1 LBD domain. 29019687 7 50 theme GluN1 1231:1235 arg1 LBD 1237:1239 the GluN1 LBD 1227:1239 the GluN1 LBD of NMDARs 1227:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 4 51 theme glycans 757:763 arg1 behavior 745:752 the behavior 741:752 the behavior of glycans on NMDARs 741:773 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 1 52 gly glycosylated 262:273 arg1 receptors 167:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors 139:175 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 1 52 gly glycosylated 262:273 arg1 receptors 191:199 key neuronal receptors 178:199 key neuronal receptors playing the central role in learning and memory 178:247 N-Methyl-d-aspartate (NMDA) receptors, key neuronal receptors playing the central role in learning and memory, are heavily glycosylated in vivo. 29019687 4 53 theme long 662:665 arg1 μs 659:660 aggregate 86.5 μs long 644:665 aggregate 86.5 μs long 644:665 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 7 54 theme changes 1205:1211 arg1 scales 1180:1185 time scales 1175:1185 time scales of conformational changes in glycans on the GluN1 LBD of NMDARs 1175:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 7 55 theme NMDARs 1244:1249 arg1 LBD 1237:1239 the GluN1 LBD 1227:1239 the GluN1 LBD of NMDARs 1227:1249 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 4 56 from behavior 745:752 arg1 NMDARs 768:773 NMDARs 768:773 NMDARs 768:773 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 9 57 theme computational 1517:1529 arg1 models 1531:1536 computational models 1517:1536 computational models of NMDARs 1517:1546 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 7 58 from nanoseconds 1262:1272 arg1 range 1251:1255 range 1251:1255 range 1251:1255 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 9 59 theme NMDARs 1505:1510 arg1 part 1497:1500 an integral part 1485:1500 an integral part of NMDARs 1485:1510 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 9 59 theme NMDARs 1505:1510 arg1 glycans 1473:1479 glycans 1473:1479 glycans 1473:1479 Thus, glycans are an integral part of NMDARs, and computational models of NMDARs should include glycans to faithfully represent the structure and the dynamics of these receptors. 29019687 2 60 attach attached 384:391 arg1 them 396:399 them 396:399 them 396:399 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 2 60 attach attached 384:391 arg2 glycans 376:382 glycans 376:382 glycans attached to them 376:399 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 4 61 theme NMDAR 720:724 arg1 subunit 705:711 the GluN1 subunit 695:711 the GluN1 subunit of the NMDAR 695:724 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 29019687 3 62 theme binding 462:468 arg1 LBD 478:480 LBD 478:480 LBD 478:480 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 62 theme binding 462:468 arg1 domain 470:475 the ligand binding domain 451:475 the ligand binding domain (LBD) of NMDA receptors (NMDARs) 451:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 62 theme binding 462:468 arg1 receptors 491:499 NMDA receptors 486:499 NMDA receptors (NMDARs) 486:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 6 63 theme slowest-time 938:949 arg1 motions 957:963 The slowest-time scale motions 934:963 The slowest-time scale motions that we detected in various glycans in some cases 934:1013 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 5 64 theme glycans 865:871 arg1 dynamics 853:860 the dynamics 849:860 the dynamics of glycans at different locations on NMDARs 849:904 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 5 64 theme glycans 865:871 arg1 different 923:931 different 923:931 different 923:931 Though all glycans in our simulations were structurally the same (Man5), the dynamics of glycans at different locations on NMDARs was surprisingly different. 29019687 0 65 theme Receptors 63:71 arg1 Domain 48:53 Ligand-Binding Domain 33:53 Ligand-Binding Domain of NMDA Receptors 33:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 0 65 theme Receptors 63:71 arg1 Receptors 63:71 NMDA Receptors 58:71 NMDA Receptors 58:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 6 66 theme scale 951:955 arg1 motions 957:963 The slowest-time scale motions 934:963 The slowest-time scale motions that we detected in various glycans in some cases 934:1013 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 3 67 theme NMDAR 569:573 arg1 coagonists 575:584 NMDAR coagonists 569:584 NMDAR coagonists 569:584 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 7 68 from changes 1205:1211 arg1 glycans 1216:1222 glycans 1216:1222 glycans 1216:1222 We predict that time scales of conformational changes in glycans on the GluN1 LBD of NMDARs range from nanoseconds to at least hundreds of microseconds. 29019687 2 69 theme physiological 349:361 arg1 relevance 363:371 physiological relevance 349:371 physiological relevance 349:371 Astonishingly little is known about the structure, dynamics, and physiological relevance of glycans attached to them. 29019687 3 70 theme certain 432:438 arg1 glycans 440:446 certain glycans 432:446 certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) 432:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 70 theme certain 432:438 arg1 potentiators 538:549 intramolecular potentiators 523:549 intramolecular potentiators 523:549 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 0 71 theme NMDA 58:61 arg1 Receptors 63:71 NMDA Receptors 58:71 NMDA Receptors 58:71 Simulated Dynamics of Glycans on Ligand-Binding Domain of NMDA Receptors Reveals Strong Dynamic Coupling between Glycans and Protein Core. 29019687 6 72 theme parts 1045:1049 arg1 flipping 1033:1040 a flipping 1031:1040 a flipping of parts of glycans relative to each other 1031:1083 The slowest-time scale motions that we detected in various glycans in some cases corresponded to a flipping of parts of glycans relative to each other, while in other cases they reduced to a head-to-tail bending of a glycan. 29019687 3 73 theme coagonists 575:584 arg1 EC50 561:564 EC50 561:564 EC50 of NMDAR coagonists 561:584 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 74 theme receptors 491:499 arg1 LBD 478:480 LBD 478:480 LBD 478:480 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 74 theme receptors 491:499 arg1 domain 470:475 the ligand binding domain 451:475 the ligand binding domain (LBD) of NMDA receptors (NMDARs) 451:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 3 74 theme receptors 491:499 arg1 receptors 491:499 NMDA receptors 486:499 NMDA receptors (NMDARs) 486:508 We recently demonstrated that certain glycans on the ligand binding domain (LBD) of NMDA receptors (NMDARs) can serve as intramolecular potentiators, changing EC50 of NMDAR coagonists. 29019687 4 75 gly glycosylated 675:686 arg1 LBD 688:690 the glycosylated LBD 671:690 the glycosylated LBD of the GluN1 subunit of the NMDAR 671:724 In this work, we use molecular dynamics trajectories, in aggregate 86.5 μs long, of the glycosylated LBD of the GluN1 subunit of the NMDAR to investigate the behavior of glycans on NMDARs. 28888453 0 0 theme antioxidant 75:85 arg1 properties 87:96 structural, functional and antioxidant properties 48:96 structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 48:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 1 1 theme functional 308:317 arg1 properties 335:344 structural, functional and antioxidant properties 296:344 structural, functional and antioxidant properties of glycated protein 296:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 6 2 from 420nm 984:988 arg1 reduction 923:931 the reduction 919:931 the reduction of free amino groups 919:952 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 2 from 420nm 984:988 arg1 absorbance 970:979 absorbance 970:979 absorbance at 420nm more than CH (p<0.05) 970:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 2 from 420nm 984:988 arg1 increase 958:965 increase 958:965 increase in absorbance at 420nm more than CH (p<0.05) 958:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 3 dep more 990:993 arg1 p<0.05 1004:1009 p<0.05 1004:1009 p<0.05 1004:1009 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 5 4 dep structural 710:719 arg1 hydrophobicity 730:743 surface hydrophobicity 722:743 surface hydrophobicity 722:743 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 5 4 dep structural 710:719 arg1 dichroism 758:766 circular dichroism 749:766 circular dichroism 749:766 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 4 5 theme K-means 576:582 arg1 clustering 584:593 K-means clustering 576:593 K-means clustering 576:593 Afterwards, K-means clustering was applied to classify all treatments into three levels of glycation (low, medium, high). 28888453 5 6 theme circular 749:756 arg1 dichroism 758:766 circular dichroism 749:766 circular dichroism 749:766 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 7 7 theme emulsion 1061:1068 arg1 activity 1070:1077 emulsion activity 1061:1077 emulsion activity 1061:1077 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 1 8 theme present 176:182 arg1 study 184:188 present study 176:188 present study 176:188 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 9 theme antioxidant 323:333 arg1 properties 335:344 structural, functional and antioxidant properties 296:344 structural, functional and antioxidant properties of glycated protein 296:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 6 10 theme groups 947:952 arg1 reduction 923:931 the reduction 919:931 the reduction of free amino groups 919:952 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 10 theme groups 947:952 arg1 increase 958:965 increase 958:965 increase in absorbance at 420nm more than CH (p<0.05) 958:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 5 11 dep antioxidant 785:795 arg1 scavenging 811:820 DPPH radical scavenging 798:820 DPPH radical scavenging 798:820 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 7 12 theme foam 1083:1086 arg1 capacity 1088:1095 foam capacity 1083:1095 foam capacity 1083:1095 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 0 13 theme serum 108:112 arg1 albumin-maltodextrin 114:133 bovine serum albumin-maltodextrin 101:133 bovine serum albumin-maltodextrin 101:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 6 14 from increase 958:965 arg1 420nm 984:988 420nm 984:988 420nm more than CH (p<0.05) 984:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 14 from increase 958:965 arg1 absorbance 970:979 absorbance 970:979 absorbance at 420nm more than CH (p<0.05) 970:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 4 15 theme glycation 655:663 arg1 levels 645:650 three levels 639:650 three levels of glycation (low, medium, high) 639:683 Afterwards, K-means clustering was applied to classify all treatments into three levels of glycation (low, medium, high). 28888453 0 16 theme bovine 101:106 arg1 albumin-maltodextrin 114:133 bovine serum albumin-maltodextrin 101:133 bovine serum albumin-maltodextrin 101:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 1 17 theme glycated 349:356 arg1 protein 358:364 glycated protein 349:364 glycated protein 349:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 2 18 theme albumin-maltodextrin 447:466 arg1 system 468:473 bovine serum albumin-maltodextrin system 434:473 bovine serum albumin-maltodextrin system 434:473 The glycation process was carried out at 90°C, in bovine serum albumin-maltodextrin system. 28888453 7 19 theme other 1020:1024 arg1 hand 1026:1029 the other hand 1016:1029 the other hand 1016:1029 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 7 20 theme MH 1118:1119 arg1 heating 1121:1127 MH heating 1118:1127 MH heating which could be attributed to different structural changes of proteins under MH and CH 1118:1213 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 5 21 theme structural 710:719 arg1 properties 823:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 1 22 theme protein 358:364 arg1 progression 280:290 Maillard reaction (MR) progression 257:290 Maillard reaction (MR) progression 257:290 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 22 theme protein 358:364 arg1 properties 335:344 structural, functional and antioxidant properties 296:344 structural, functional and antioxidant properties of glycated protein 296:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 5 23 theme surface 722:728 arg1 hydrophobicity 730:743 surface hydrophobicity 722:743 surface hydrophobicity 722:743 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 5 24 theme antioxidant 785:795 arg1 properties 823:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 1 25 theme Maillard 257:264 arg1 MR 276:277 MR 276:277 MR 276:277 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 25 theme Maillard 257:264 arg1 reaction 266:273 Maillard reaction 257:273 Maillard reaction (MR) progression 257:290 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 7 26 theme higher 1032:1037 arg1 values 1039:1044 higher values 1032:1044 higher values in solubility, emulsion activity and foam capacity 1032:1095 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 5 27 theme chosen 855:860 arg1 treatments 862:871 chosen treatments 855:871 chosen treatments 855:871 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 1 28 theme reaction 266:273 arg1 progression 280:290 Maillard reaction (MR) progression 257:290 Maillard reaction (MR) progression 257:290 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 29 theme conventional 201:212 arg1 heating 214:220 conventional heating 201:220 conventional heating (CH) 201:225 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 29 theme conventional 201:212 arg1 CH 223:224 CH 223:224 CH 223:224 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 0 30 theme microwave 10:18 arg1 heating 37:43 microwave and conventional heating 10:43 microwave and conventional heating 10:43 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 6 31 theme free 936:939 arg1 groups 947:952 free amino groups 936:952 free amino groups 936:952 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 0 32 theme albumin-maltodextrin 114:133 arg1 properties 87:96 structural, functional and antioxidant properties 48:96 structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 48:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 3 33 theme glycation 490:498 arg1 extent 480:485 The extent 476:485 The extent of glycation 476:498 The extent of glycation was assessed by ortho-phthaldialdehyde and absorbance at 420nm. 28888453 1 34 theme heating 214:220 arg1 effect 191:196 effect 191:196 effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein 191:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 0 35 theme heating 37:43 arg1 Effect 0:5 Effect 0:5 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 0:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 7 36 from values 1039:1044 arg1 solubility 1049:1058 solubility 1049:1058 solubility 1049:1058 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 7 36 from values 1039:1044 arg1 activity 1070:1077 emulsion activity 1061:1077 emulsion activity 1061:1077 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 7 36 from values 1039:1044 arg1 capacity 1088:1095 foam capacity 1083:1095 foam capacity 1083:1095 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 0 37 theme Maillard 154:161 arg1 reaction 163:170 Maillard reaction 154:170 Maillard reaction 154:170 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 4 38 dep levels 645:650 arg1 high 679:682 high 679:682 high 679:682 Afterwards, K-means clustering was applied to classify all treatments into three levels of glycation (low, medium, high). 28888453 4 38 dep levels 645:650 arg1 medium 671:676 medium 671:676 medium 671:676 Afterwards, K-means clustering was applied to classify all treatments into three levels of glycation (low, medium, high). 28888453 4 38 dep levels 645:650 arg1 low 666:668 low 666:668 low 666:668 Afterwards, K-means clustering was applied to classify all treatments into three levels of glycation (low, medium, high). 28888453 0 39 theme conventional 24:35 arg1 heating 37:43 microwave and conventional heating 10:43 microwave and conventional heating 10:43 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 2 40 theme serum 441:445 arg1 system 468:473 bovine serum albumin-maltodextrin system 434:473 bovine serum albumin-maltodextrin system 434:473 The glycation process was carried out at 90°C, in bovine serum albumin-maltodextrin system. 28888453 2 41 theme glycation 388:396 arg1 process 398:404 The glycation process 384:404 The glycation process 384:404 The glycation process was carried out at 90°C, in bovine serum albumin-maltodextrin system. 28888453 0 42 theme structural 48:57 arg1 properties 87:96 structural, functional and antioxidant properties 48:96 structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 48:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 5 43 theme functional 770:779 arg1 properties 823:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties 710:832 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 6 44 theme amino 941:945 arg1 groups 947:952 free amino groups 936:952 free amino groups 936:952 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 0 45 from Effect 0:5 arg1 properties 87:96 structural, functional and antioxidant properties 48:96 structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 48:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 7 46 theme different 1158:1166 arg1 changes 1179:1185 different structural changes 1158:1185 different structural changes of proteins under MH and CH 1158:1213 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 1 47 theme structural 296:305 arg1 properties 335:344 structural, functional and antioxidant properties 296:344 structural, functional and antioxidant properties of glycated protein 296:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 6 48 from reduction 923:931 arg1 420nm 984:988 420nm 984:988 420nm more than CH (p<0.05) 984:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 6 48 from reduction 923:931 arg1 absorbance 970:979 absorbance 970:979 absorbance at 420nm more than CH (p<0.05) 970:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 1 49 from effect 191:196 arg1 progression 280:290 Maillard reaction (MR) progression 257:290 Maillard reaction (MR) progression 257:290 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 49 from effect 191:196 arg1 properties 335:344 structural, functional and antioxidant properties 296:344 structural, functional and antioxidant properties of glycated protein 296:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 6 50 theme more 990:993 arg1 420nm 984:988 420nm 984:988 420nm more than CH (p<0.05) 984:1010 Results showed that MH was more effective on the reduction of free amino groups and increase in absorbance at 420nm more than CH (p<0.05). 28888453 0 51 theme functional 60:69 arg1 properties 87:96 structural, functional and antioxidant properties 48:96 structural, functional and antioxidant properties of bovine serum albumin-maltodextrin 48:133 Effect of microwave and conventional heating on structural, functional and antioxidant properties of bovine serum albumin-maltodextrin conjugates through Maillard reaction. 28888453 5 52 theme radical 803:809 arg1 scavenging 811:820 DPPH radical scavenging 798:820 DPPH radical scavenging 798:820 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 7 53 theme proteins 1190:1197 arg1 changes 1179:1185 different structural changes 1158:1185 different structural changes of proteins under MH and CH 1158:1213 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 7 54 theme structural 1168:1177 arg1 changes 1179:1185 different structural changes 1158:1185 different structural changes of proteins under MH and CH 1158:1213 On the other hand, higher values in solubility, emulsion activity and foam capacity were obtained during MH heating which could be attributed to different structural changes of proteins under MH and CH. 28888453 1 55 theme microwave 231:239 arg1 MH 250:251 MH 250:251 MH 250:251 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 1 55 theme microwave 231:239 arg1 heating 241:247 microwave heating 231:247 microwave heating (MH) 231:252 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 28888453 2 56 theme bovine 434:439 arg1 system 468:473 bovine serum albumin-maltodextrin system 434:473 bovine serum albumin-maltodextrin system 434:473 The glycation process was carried out at 90°C, in bovine serum albumin-maltodextrin system. 28888453 5 57 theme DPPH 798:801 arg1 scavenging 811:820 DPPH radical scavenging 798:820 DPPH radical scavenging 798:820 Subsequently, SDS-PAGE, structural (surface hydrophobicity and circular dichroism), functional and antioxidant (DPPH radical scavenging) properties were measured within chosen treatments. 28888453 1 58 theme heating 241:247 arg1 effect 191:196 effect 191:196 effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein 191:364 In present study, effect of conventional heating (CH) and microwave heating (MH) on Maillard reaction (MR) progression and structural, functional and antioxidant properties of glycated protein was investigated. 27932526 4 0 theme plasma 925:930 arg1 N-glycans 932:940 plasma N-glycans 925:940 plasma N-glycans 925:940 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 5 1 from Study 1319:1323 arg1 individuals 1281:1291 2144 healthy middle-aged individuals 1256:1291 2144 healthy middle-aged individuals from the Leiden Longevity Study 1256:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 1 from Study 1319:1323 arg1 N-glycome 1243:1251 the total plasma N-glycome 1226:1251 the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study 1226:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 9 2 theme plasma 2360:2365 arg1 characteristics 2367:2381 important plasma characteristics 2350:2381 important plasma characteristics associated with inflammation and metabolic health 2350:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 2 theme plasma 2360:2365 arg1 sialylation 2261:2271 the sialylation 2257:2271 the sialylation of tetraantennary species 2257:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 2 theme plasma 2360:2365 arg1 size 2308:2311 the size 2304:2311 the size of high-mannose species 2304:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 2 theme plasma 2360:2365 arg1 bisection 2189:2197 bisection 2189:2197 bisection 2189:2197 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 6 3 theme pressure 1616:1623 arg1 MALDI-FTICR-MS 1625:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 5 4 theme healthy 1261:1267 arg1 individuals 1281:1291 2144 healthy middle-aged individuals 1256:1291 2144 healthy middle-aged individuals from the Leiden Longevity Study 1256:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 9 5 theme species 2329:2335 arg1 sialylation 2261:2271 the sialylation 2257:2271 the sialylation of tetraantennary species 2257:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 5 theme species 2329:2335 arg1 size 2308:2311 the size 2304:2311 the size of high-mannose species 2304:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 5 theme species 2329:2335 arg1 sialylation 2221:2231 sialylation 2221:2231 sialylation 2221:2231 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 5 theme species 2329:2335 arg1 bisection 2189:2197 bisection 2189:2197 bisection 2189:2197 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 5 theme species 2329:2335 arg1 characteristics 2367:2381 important plasma characteristics 2350:2381 important plasma characteristics associated with inflammation and metabolic health 2350:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 5 theme species 2329:2335 arg1 galactosylation 2200:2214 galactosylation 2200:2214 galactosylation 2200:2214 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 8 6 theme C-reactive 1959:1968 arg1 insulin 2088:2094 insulin 2088:2094 insulin 2088:2094 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 cholesterol 2055:2065 HDL cholesterol 2051:2065 HDL cholesterol 2051:2065 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 leptin 2030:2035 leptin 2030:2035 leptin 2030:2035 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 interleukin 1985:1995 interleukin 6 1985:1997 interleukin 6 (IL-6) 1985:2004 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 smoking 2167:2173 smoking 2167:2173 smoking 2167:2173 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 triglycerides 2068:2080 triglycerides 2068:2080 triglycerides (TG) 2068:2085 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 index 2017:2021 body mass index 2007:2021 body mass index (BMI) 2007:2027 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 transferase 2112:2122 gamma-glutamyl transferase 2097:2122 gamma-glutamyl transferase (GGT) 2097:2128 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 adiponectin 2038:2048 adiponectin 2038:2048 adiponectin 2038:2048 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 aminotransferase 2139:2154 alanine aminotransferase 2131:2154 alanine aminotransferase (ALT) 2131:2160 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 6 theme C-reactive 1959:1968 arg1 CRP 1979:1981 CRP 1979:1981 CRP 1979:1981 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 3 7 with association 620:630 arg1 levels 652:657 plasma protein levels 637:657 plasma protein levels 637:657 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 3 7 with association 620:630 arg1 processes 672:680 systemic processes 663:680 systemic processes 663:680 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 6 8 theme mode 1598:1601 arg1 MALDI-FTICR-MS 1625:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 1 9 theme abundant 215:222 arg1 Glycosylation 195:207 Glycosylation 195:207 Glycosylation 195:207 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 9 theme abundant 215:222 arg1 modification 259:270 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 5 10 used used 1107:1110 arg2 we 1099:1100 we 1099:1100 we 1099:1100 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 3 11 theme plasma 637:642 arg1 levels 652:657 plasma protein levels 637:657 plasma protein levels 637:657 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 6 12 theme negative 1585:1592 arg1 mode 1598:1601 negative ion mode 1585:1601 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 4 13 theme large-scale 693:703 arg1 studies 705:711 large-scale studies 693:711 large-scale studies (n >200) of the total plasma N-glycome 693:750 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 4 13 theme large-scale 693:703 arg1 n 714:714 n >200 714:719 n >200 714:719 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 5 14 theme matrix-assisted 1112:1126 arg1 MALDI 1157:1161 MALDI 1157:1161 MALDI 1157:1161 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 14 theme matrix-assisted 1112:1126 arg1 desorption/ionization 1134:1154 matrix-assisted laser desorption/ionization 1112:1154 matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS 1112:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 9 15 theme metabolic 2416:2424 arg1 health 2426:2431 metabolic health 2416:2431 metabolic health 2416:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 4 16 dep limited 875:881 arg1 separation 824:833 chromatographic and electrophoretic separation 788:833 chromatographic and electrophoretic separation 788:833 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 7 17 theme glycan 1700:1705 arg1 compositions 1707:1718 61 glycan compositions 1697:1718 61 glycan compositions 1697:1718 In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. 27932526 5 18 theme desorption/ionization 1134:1154 arg1 -Fourier 1163:1170 matrix-assisted laser desorption/ionization (MALDI)-Fourier 1112:1170 matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS 1112:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 3 19 theme systemic 663:670 arg1 processes 672:680 systemic processes 663:680 systemic processes 663:680 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 0 20 theme Health 187:192 arg1 Markers 149:155 Markers 149:155 Markers of Inflammation and Metabolic Health 149:192 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 8 21 theme confirming 1842:1851 arg1 associations 1859:1870 confirming known associations 1842:1870 confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS 1842:1922 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 7 22 theme relative 1670:1677 arg1 quantification 1679:1692 the relative quantification 1666:1692 the relative quantification of 61 glycan compositions 1666:1718 In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. 27932526 7 23 theme glycosylation 1796:1808 arg1 traits 1810:1815 39 glycosylation traits 1793:1815 39 glycosylation traits derived thereof 1793:1831 In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. 27932526 9 24 theme tetraantennary 2276:2289 arg1 species 2291:2297 tetraantennary species 2276:2297 tetraantennary species 2276:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 4 25 theme total 729:733 arg1 N-glycome 742:750 the total plasma N-glycome 725:750 the total plasma N-glycome 725:750 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 0 26 theme Desorption/Ionization-Fourier 66:94 arg1 Resonance-MS 120:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 5 27 theme ion 1182:1184 arg1 FTICR 1207:1211 FTICR 1207:1211 FTICR 1207:1211 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 27 theme ion 1182:1184 arg1 resonance 1196:1204 ion cyclotron resonance 1182:1204 ion cyclotron resonance (FTICR)-MS 1182:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 8 28 theme glycosylation 1875:1887 arg1 associations 1859:1870 confirming known associations 1842:1870 confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS 1842:1922 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 5 29 gly sialylation 1034:1044 arg1 species.Here 1085:1096 high-mannose type species.Here 1067:1096 high-mannose type species.Here 1067:1096 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 4 30 theme N-glycome 742:750 arg1 studies 705:711 large-scale studies 693:711 large-scale studies (n >200) of the total plasma N-glycome 693:750 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 4 30 theme N-glycome 742:750 arg1 n 714:714 n >200 714:719 n >200 714:719 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 5 31 theme Longevity 1309:1317 arg1 Study 1319:1323 the Leiden Longevity Study 1298:1323 the Leiden Longevity Study 1298:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 0 32 theme Ion 106:108 arg1 Resonance-MS 120:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 5 33 theme resonance 1196:1204 arg1 -MS 1213:1215 ion cyclotron resonance (FTICR)-MS 1182:1215 ion cyclotron resonance (FTICR)-MS 1182:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 4 34 theme separation 824:833 arg1 methods 777:783 methods 777:783 methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans 777:940 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 9 35 theme diantennary 2236:2246 arg1 species 2248:2254 diantennary species 2236:2254 diantennary species 2236:2254 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 0 36 dep Resonance-MS 120:131 arg1 Transform 96:104 Transform 96:104 Transform 96:104 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 6 37 attach released 1453:1460 arg1 backbones 1481:1489 their protein backbones 1467:1489 their protein backbones 1467:1489 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 6 37 attach released 1453:1460 arg2 N-glycans 1424:1432 N-glycans 1424:1432 N-glycans 1424:1432 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 0 38 theme Human 0:4 arg1 N-glycosylation 13:27 Human Plasma N-glycosylation 0:27 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 0:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 3 39 theme informative 526:536 arg1 sample 538:543 a complex but informative sample 512:543 a complex but informative sample like the total plasma N-glycome 512:575 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 5 40 theme total 1230:1234 arg1 N-glycome 1243:1251 the total plasma N-glycome 1226:1251 the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study 1226:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 41 theme metabolic 1372:1380 arg1 health 1382:1387 metabolic health 1372:1387 metabolic health 1372:1387 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 3 42 theme complex 514:520 arg1 sample 538:543 a complex but informative sample 512:543 a complex but informative sample like the total plasma N-glycome 512:575 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 6 43 theme protein 1473:1479 arg1 backbones 1481:1489 their protein backbones 1467:1489 their protein backbones 1467:1489 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 5 44 theme high-mannose 1067:1078 arg1 species.Here 1085:1096 high-mannose type species.Here 1067:1096 high-mannose type species.Here 1067:1096 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 8 45 theme body 2007:2010 arg1 index 2017:2021 body mass index 2007:2021 body mass index (BMI) 2007:2027 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 45 theme body 2007:2010 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 45 theme body 2007:2010 arg1 BMI 2024:2026 BMI 2024:2026 BMI 2024:2026 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 3 46 theme total 554:558 arg1 N-glycome 567:575 the total plasma N-glycome 550:575 the total plasma N-glycome 550:575 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 0 47 theme Matrix-Assisted 44:58 arg1 Resonance-MS 120:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 5 48 theme species.Here 1085:1096 arg1 others 1013:1018 others 1013:1018 others 1013:1018 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 48 theme species.Here 1085:1096 arg1 antennarity 1021:1031 antennarity 1021:1031 antennarity 1021:1031 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 48 theme species.Here 1085:1096 arg1 identity 1055:1062 the identity 1051:1062 the identity of high-mannose type species.Here 1051:1096 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 48 theme species.Here 1085:1096 arg1 sialylation 1034:1044 sialylation 1034:1044 sialylation 1034:1044 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 49 theme middle-aged 1269:1279 arg1 individuals 1281:1291 2144 healthy middle-aged individuals 1256:1291 2144 healthy middle-aged individuals from the Leiden Longevity Study 1256:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 0 50 dep N-glycosylation 13:27 arg1 Analyzed 32:39 Analyzed 32:39 Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 32:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 2 51 dep not 331:333 arg1 template-defined 335:350 template-defined 335:350 template-defined 335:350 Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. 27932526 2 52 theme high-potential 425:438 arg1 biomarker 440:448 a high-potential biomarker 423:448 a high-potential biomarker for disease and patient stratification 423:487 Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. 27932526 1 53 mod modification 259:270 arg1 importance 275:284 importance 275:284 importance 275:284 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 53 mod modification 259:270 arg3 protein 251:257 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 8 54 theme novel 1935:1939 arg1 associations 1941:1952 novel associations 1935:1952 novel associations 1935:1952 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 9 55 theme important 2350:2358 arg1 characteristics 2367:2381 important plasma characteristics 2350:2381 important plasma characteristics associated with inflammation and metabolic health 2350:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 55 theme important 2350:2358 arg1 sialylation 2261:2271 the sialylation 2257:2271 the sialylation of tetraantennary species 2257:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 55 theme important 2350:2358 arg1 size 2308:2311 the size 2304:2311 the size of high-mannose species 2304:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 55 theme important 2350:2358 arg1 bisection 2189:2197 bisection 2189:2197 bisection 2189:2197 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 6 56 theme reducing 1507:1514 arg1 end 1516:1518 the reducing end 1503:1518 the reducing end 1503:1518 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 4 57 theme N-glycans 932:940 arg1 complexity 911:920 the structural complexity 896:920 the structural complexity of plasma N-glycans 896:940 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 2 58 theme organism 407:414 arg1 state 395:399 the biological state 380:399 the biological state of an organism 380:414 Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. 27932526 6 59 theme intermediate 1603:1614 arg1 MALDI-FTICR-MS 1625:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 2 60 theme biological 384:393 arg1 state 395:399 the biological state 380:399 the biological state of an organism 380:414 Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. 27932526 6 61 theme ion 1594:1596 arg1 mode 1598:1601 negative ion mode 1585:1601 negative ion mode intermediate pressure MALDI-FTICR-MS 1585:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 1 62 theme co- 224:226 arg1 Glycosylation 195:207 Glycosylation 195:207 Glycosylation 195:207 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 62 theme co- 224:226 arg1 modification 259:270 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 7 63 theme compositions 1707:1718 arg1 quantification 1679:1692 the relative quantification 1666:1692 the relative quantification of 61 glycan compositions 1666:1718 In doing so, we achieved the relative quantification of 61 glycan compositions, ranging from Hex4HexNAc2 to Hex7HexNAc6dHex1Neu5Ac4, as well as that of 39 glycosylation traits derived thereof. 27932526 4 64 theme structural 900:909 arg1 complexity 911:920 the structural complexity 896:920 the structural complexity of plasma N-glycans 896:940 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 0 65 theme Inflammation 160:171 arg1 Markers 149:155 Markers 149:155 Markers of Inflammation and Metabolic Health 149:192 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 3 66 theme baseline 611:618 arg1 association 620:630 its baseline association 607:630 its baseline association with plasma protein levels and systemic processes 607:680 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 1 67 theme post-translational 232:249 arg1 Glycosylation 195:207 Glycosylation 195:207 Glycosylation 195:207 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 67 theme post-translational 232:249 arg1 modification 259:270 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 68 dep protein 289:295 arg1 processing 297:306 processing 297:306 processing 297:306 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 8 69 with glycosylation 1875:1887 arg1 sex 1902:1904 sex 1902:1904 sex 1902:1904 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 69 with glycosylation 1875:1887 arg1 age 1894:1896 age 1894:1896 age 1894:1896 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 5 70 from N-glycome 1243:1251 arg1 Study 1319:1323 the Leiden Longevity Study 1298:1323 the Leiden Longevity Study 1298:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 71 theme laser 1128:1132 arg1 MALDI 1157:1161 MALDI 1157:1161 MALDI 1157:1161 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 71 theme laser 1128:1132 arg1 desorption/ionization 1134:1154 matrix-assisted laser desorption/ionization 1112:1154 matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS 1112:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 3 72 theme protein 644:650 arg1 levels 652:657 plasma protein levels 637:657 plasma protein levels 637:657 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 5 73 theme additional 980:989 arg1 information 991:1001 additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation 980:1404 additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation 980:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 9 74 gly sialylation 2221:2231 arg1 species 2291:2297 tetraantennary species 2276:2297 tetraantennary species 2276:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 74 gly sialylation 2221:2231 arg1 species 2329:2335 high-mannose species 2316:2335 high-mannose species 2316:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 74 gly sialylation 2221:2231 arg1 species 2248:2254 diantennary species 2236:2254 diantennary species 2236:2254 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 5 75 theme association 1335:1345 arg1 analysis 1347:1354 association analysis 1335:1354 association analysis with markers of metabolic health and inflammation 1335:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 8 76 theme mass 2012:2015 arg1 index 2017:2021 body mass index 2007:2021 body mass index (BMI) 2007:2027 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 76 theme mass 2012:2015 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 76 theme mass 2012:2015 arg1 BMI 2024:2026 BMI 2024:2026 BMI 2024:2026 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 77 theme known 1853:1857 arg1 associations 1859:1870 confirming known associations 1842:1870 confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS 1842:1922 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 5 78 contain has 946:948 arg1 MS 943:944 MS 943:944 MS 943:944 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 78 contain has 946:948 arg2 opportunity 954:964 the opportunity 950:964 the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation 950:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 1 79 theme importance 275:284 arg1 Glycosylation 195:207 Glycosylation 195:207 Glycosylation 195:207 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 79 theme importance 275:284 arg1 modification 259:270 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 5 80 theme Leiden 1302:1307 arg1 Study 1319:1323 the Leiden Longevity Study 1298:1323 the Leiden Longevity Study 1298:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 0 81 theme Cyclotron 110:118 arg1 Resonance-MS 120:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 5 82 theme cyclotron 1186:1194 arg1 FTICR 1207:1211 FTICR 1207:1211 FTICR 1207:1211 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 82 theme cyclotron 1186:1194 arg1 resonance 1196:1204 ion cyclotron resonance 1182:1204 ion cyclotron resonance (FTICR)-MS 1182:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 4 83 theme plasma 735:740 arg1 N-glycome 742:750 the total plasma N-glycome 725:750 the total plasma N-glycome 725:750 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 4 84 theme electrophoretic 808:822 arg1 separation 824:833 chromatographic and electrophoretic separation 788:833 chromatographic and electrophoretic separation 788:833 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 5 85 theme health 1382:1387 arg1 markers 1361:1367 markers 1361:1367 markers of metabolic health and inflammation 1361:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 9 86 theme species 2248:2254 arg1 sialylation 2261:2271 the sialylation 2257:2271 the sialylation of tetraantennary species 2257:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 86 theme species 2248:2254 arg1 size 2308:2311 the size 2304:2311 the size of high-mannose species 2304:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 86 theme species 2248:2254 arg1 sialylation 2221:2231 sialylation 2221:2231 sialylation 2221:2231 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 86 theme species 2248:2254 arg1 bisection 2189:2197 bisection 2189:2197 bisection 2189:2197 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 86 theme species 2248:2254 arg1 characteristics 2367:2381 important plasma characteristics 2350:2381 important plasma characteristics associated with inflammation and metabolic health 2350:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 86 theme species 2248:2254 arg1 galactosylation 2200:2214 galactosylation 2200:2214 galactosylation 2200:2214 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 5 87 with analysis 1347:1354 arg1 markers 1361:1367 markers 1361:1367 markers of metabolic health and inflammation 1361:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 1 88 theme protein 251:257 arg1 Glycosylation 195:207 Glycosylation 195:207 Glycosylation 195:207 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 1 88 theme protein 251:257 arg1 modification 259:270 an abundant co- and post-translational protein modification 212:270 an abundant co- and post-translational protein modification of importance to protein processing and activity 212:319 Glycosylation is an abundant co- and post-translational protein modification of importance to protein processing and activity. 27932526 5 89 theme inflammation 1393:1404 arg1 markers 1361:1367 markers 1361:1367 markers of metabolic health and inflammation 1361:1404 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 0 90 theme Plasma 6:11 arg1 N-glycosylation 13:27 Human Plasma N-glycosylation 0:27 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 0:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 6 91 dep released 1453:1460 arg1 analyzed 1573:1580 analyzed 1573:1580 analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS 1573:1638 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27932526 0 92 theme Metabolic 177:185 arg1 Health 187:192 Metabolic Health 177:192 Metabolic Health 177:192 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 8 93 theme HDL 2051:2053 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 93 theme HDL 2051:2053 arg1 cholesterol 2055:2065 HDL cholesterol 2051:2065 HDL cholesterol 2051:2065 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 94 theme gamma-glutamyl 2097:2110 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 94 theme gamma-glutamyl 2097:2110 arg1 GGT 2125:2127 GGT 2125:2127 GGT 2125:2127 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 94 theme gamma-glutamyl 2097:2110 arg1 transferase 2112:2122 gamma-glutamyl transferase 2097:2122 gamma-glutamyl transferase (GGT) 2097:2128 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 4 95 theme chromatographic 788:802 arg1 separation 824:833 chromatographic and electrophoretic separation 788:833 chromatographic and electrophoretic separation 788:833 Thus far, large-scale studies (n >200) of the total plasma N-glycome have been performed with methods of chromatographic and electrophoretic separation, which, although being informative, are limited in resolving the structural complexity of plasma N-glycans. 27932526 9 96 theme species 2291:2297 arg1 sialylation 2261:2271 the sialylation 2257:2271 the sialylation of tetraantennary species 2257:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 96 theme species 2291:2297 arg1 size 2308:2311 the size 2304:2311 the size of high-mannose species 2304:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 96 theme species 2291:2297 arg1 sialylation 2221:2231 sialylation 2221:2231 sialylation 2221:2231 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 96 theme species 2291:2297 arg1 bisection 2189:2197 bisection 2189:2197 bisection 2189:2197 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 96 theme species 2291:2297 arg1 characteristics 2367:2381 important plasma characteristics 2350:2381 important plasma characteristics associated with inflammation and metabolic health 2350:2431 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 96 theme species 2291:2297 arg1 galactosylation 2200:2214 galactosylation 2200:2214 galactosylation 2200:2214 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 5 97 theme plasma 1236:1241 arg1 N-glycome 1243:1251 the total plasma N-glycome 1226:1251 the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study 1226:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 5 98 dep -Fourier 1163:1170 arg1 transform 1172:1180 transform 1172:1180 transform ion cyclotron resonance (FTICR)-MS 1172:1215 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 0 99 theme Laser 60:64 arg1 Resonance-MS 120:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS 44:131 Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health. 27932526 9 100 gly sialylation 2261:2271 arg1 species 2291:2297 tetraantennary species 2276:2297 tetraantennary species 2276:2297 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 100 gly sialylation 2261:2271 arg1 species 2329:2335 high-mannose species 2316:2335 high-mannose species 2316:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 9 100 gly sialylation 2261:2271 arg1 species 2248:2254 diantennary species 2236:2254 diantennary species 2236:2254 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 5 101 theme type 1080:1083 arg1 species.Here 1085:1096 high-mannose type species.Here 1067:1096 high-mannose type species.Here 1067:1096 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 3 102 theme plasma 560:565 arg1 N-glycome 567:575 the total plasma N-glycome 550:575 the total plasma N-glycome 550:575 However, to interpret a complex but informative sample like the total plasma N-glycome, it is important to establish its baseline association with plasma protein levels and systemic processes. 27932526 8 103 theme alanine 2131:2137 arg1 protein 1970:1976 C-reactive protein 1959:1976 C-reactive protein (CRP) 1959:1982 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 103 theme alanine 2131:2137 arg1 ALT 2157:2159 ALT 2157:2159 ALT 2157:2159 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 8 103 theme alanine 2131:2137 arg1 aminotransferase 2139:2154 alanine aminotransferase 2131:2154 alanine aminotransferase (ALT) 2131:2160 Next to confirming known associations of glycosylation with age and sex by MALDI-FTICR-MS, we report novel associations with C-reactive protein (CRP), interleukin 6 (IL-6), body mass index (BMI), leptin, adiponectin, HDL cholesterol, triglycerides (TG), insulin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and smoking. 27932526 9 104 theme high-mannose 2316:2327 arg1 species 2329:2335 high-mannose species 2316:2335 high-mannose species 2316:2335 Overall, the bisection, galactosylation, and sialylation of diantennary species, the sialylation of tetraantennary species, and the size of high-mannose species proved to be important plasma characteristics associated with inflammation and metabolic health. 27932526 5 105 theme individuals 1281:1291 arg1 N-glycome 1243:1251 the total plasma N-glycome 1226:1251 the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study 1226:1323 MS has the opportunity to contribute additional information on, among others, antennarity, sialylation, and the identity of high-mannose type species.Here, we have used matrix-assisted laser desorption/ionization (MALDI)-Fourier transform ion cyclotron resonance (FTICR)-MS to study the total plasma N-glycome of 2144 healthy middle-aged individuals from the Leiden Longevity Study, to allow association analysis with markers of metabolic health and inflammation. 27932526 2 106 dep disease 454:460 arg1 stratification 474:487 stratification 474:487 stratification 474:487 Although not template-defined, glycosylation does reflect the biological state of an organism and is a high-potential biomarker for disease and patient stratification. 27932526 6 107 theme 2-aminobenzoic 1525:1538 arg1 acid 1540:1543 2-aminobenzoic acid 1525:1543 2-aminobenzoic acid 1525:1543 To achieve this, N-glycans were enzymatically released from their protein backbones, labeled at the reducing end with 2-aminobenzoic acid, and following purification analyzed by negative ion mode intermediate pressure MALDI-FTICR-MS. 27751954 8 0 theme first 1577:1581 arg1 time 1583:1586 the first time 1573:1586 the first time in a parasitic nematode 1573:1610 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 2 1 theme sexual 352:357 arg1 organism 359:366 an obligate sexual organism 340:366 an obligate sexual organism 340:366 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 2 1 theme sexual 352:357 arg1 dentatum 328:335 the parasitic O. dentatum 311:335 the parasitic O. dentatum 311:335 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 7 2 located observed 1364:1371 arg1 studies 1389:1395 our previous studies 1376:1395 our previous studies on other nematodes 1376:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 7 2 located observed 1364:1371 arg2 motifs 1353:1358 extended HexNAc2-4PC2-4 motifs 1329:1358 extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1329:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 2 3 theme C. 299:300 arg1 elegans 302:308 the non-parasitic C. elegans 281:308 the non-parasitic C. elegans 281:308 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 7 4 theme other 1400:1404 arg1 nematodes 1406:1414 other nematodes 1400:1414 other nematodes 1400:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 5 5 theme wide 843:846 arg1 range 848:852 a wide range 841:852 a wide range of core fucosylated N-glycans with up to three fucose residues 841:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 3 6 theme L3 572:573 arg1 larvae 582:587 L3 and L4 larvae 572:587 larvae 582:587 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 1 7 theme related 203:209 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 7 theme related 203:209 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 6 8 theme paucimannosidic 947:961 arg1 structures 963:972 simple methylated paucimannosidic structures 929:972 simple methylated paucimannosidic structures 929:972 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 5 9 theme female 820:825 arg1 worms 827:831 the male and female worms 807:831 the male and female worms 807:831 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 6 10 theme simple 929:934 arg1 structures 963:972 simple methylated paucimannosidic structures 929:972 simple methylated paucimannosidic structures 929:972 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 7 11 theme paucimannosidic 1122:1136 arg1 glycans 1138:1144 the fucosylated paucimannosidic glycans 1106:1144 the fucosylated paucimannosidic glycans 1106:1144 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 1 12 theme organism 224:231 arg1 elegans 248:254 the model organism Caenorhabditis elegans 214:254 the model organism Caenorhabditis elegans 214:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 5 13 gly fucosylated 862:872 arg1 N-glycans 874:882 core fucosylated N-glycans 857:882 core fucosylated N-glycans with up to three fucose residues 857:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 4 14 theme core 687:690 arg1 RP-HPLC 692:698 fused core RP-HPLC 681:698 fused core RP-HPLC 681:698 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 8 15 from time 1583:1586 arg1 nematode 1603:1610 a parasitic nematode 1591:1610 a parasitic nematode 1591:1610 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 8 16 dep O. 1529:1530 arg1 dentatum 1532:1539 O. dentatum 1529:1539 O. dentatum 1529:1539 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 3 17 theme L4 579:580 arg1 larvae 582:587 L3 and L4 larvae 572:587 larvae 582:587 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 3 18 dep O. 544:545 arg1 dentatum 547:554 O. dentatum 544:554 O. dentatum 544:554 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 5 19 dep RESULTS 701:707 arg1 more 750:753 more 750:753 more 750:753 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 5 19 dep RESULTS 701:707 arg1 simpler 738:744 simpler 738:744 simpler 738:744 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 5 19 dep RESULTS 701:707 arg1 N-glycome 724:732 the L3 N-glycome 717:732 the L3 N-glycome 717:732 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 8 20 theme class 1457:1461 arg1 nematodes 1465:1473 class V nematodes 1457:1473 class V nematodes 1457:1473 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 7 21 theme extended 1329:1336 arg1 motifs 1353:1358 extended HexNAc2-4PC2-4 motifs 1329:1358 extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1329:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 3 22 dep male 528:531 arg1 O. 544:545 O. 544:545 O. 544:545 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 5 23 theme fucose 901:906 arg1 residues 908:915 up to three fucose residues 889:915 up to three fucose residues 889:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 24 from studies 1389:1395 arg1 nematodes 1406:1414 other nematodes 1400:1414 other nematodes 1400:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 9 25 theme methylated 1706:1715 arg1 SIGNIFICANCE 1685:1696 GENERAL SIGNIFICANCE 1677:1696 GENERAL SIGNIFICANCE Unusual methylated 1677:1715 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 8 26 theme nematodes 1465:1473 arg1 tendency 1445:1452 The interspecies tendency 1428:1452 The interspecies tendency of class V nematodes 1428:1473 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 7 27 theme glycans 1316:1322 arg1 range 1280:1284 a wide range 1273:1284 a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1273:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 0 28 theme parasite 60:67 arg1 dentatum 85:92 the porcine parasite Oesophagostomum dentatum 48:92 the porcine parasite Oesophagostomum dentatum 48:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 1 29 theme class 171:175 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 29 theme class 171:175 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 7 30 theme glycans 1138:1144 arg1 glycans 1138:1144 the fucosylated paucimannosidic glycans 1106:1144 the fucosylated paucimannosidic glycans 1106:1144 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 7 30 theme glycans 1138:1144 arg1 many 1098:1101 many 1098:1101 many 1098:1101 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 5 31 theme fucosylated 862:872 arg1 N-glycans 874:882 core fucosylated N-glycans 857:882 core fucosylated N-glycans with up to three fucose residues 857:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 32 from species 1194:1200 arg1 examples 1165:1172 examples 1165:1172 examples from other nematode species 1165:1200 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 9 33 theme core 1718:1721 arg1 fucosylated 1723:1733 core fucosylated 1718:1733 core fucosylated 1718:1733 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 5 34 with N-glycans 874:882 arg1 residues 908:915 up to three fucose residues 889:915 up to three fucose residues 889:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 1 35 dep BACKGROUND 95:104 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 35 dep BACKGROUND 95:104 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 0 36 theme dentatum 85:92 arg1 Gender 0:5 Gender 0:5 Gender 0:5 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 0 36 theme dentatum 85:92 arg1 N-glycomes 34:43 developmental specific N-glycomes 11:43 developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum 11:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 1 37 theme V 177:177 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 37 theme V 177:177 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 5 38 dep three 895:899 arg1 to 892:893 to 892:893 to 892:893 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 39 theme other 1179:1183 arg1 species 1194:1200 other nematode species 1179:1200 other nematode species 1179:1200 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 5 40 theme L3 721:722 arg1 simpler 738:744 simpler 738:744 simpler 738:744 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 5 40 theme L3 721:722 arg1 N-glycome 724:732 the L3 N-glycome 717:732 the L3 N-glycome 717:732 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 41 theme phosphorylcholine-modified 1289:1314 arg1 glycans 1316:1322 phosphorylcholine-modified glycans 1289:1322 phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1289:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 0 42 theme Oesophagostomum 69:83 arg1 dentatum 85:92 the porcine parasite Oesophagostomum dentatum 48:92 the porcine parasite Oesophagostomum dentatum 48:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 6 43 gly fucosylated 1021:1031 arg1 glycans 1033:1039 fucosylated glycans 1021:1039 fucosylated glycans 1021:1039 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 3 44 used used 497:500 arg2 METHODS 443:449 METHODS 443:449 METHODS Different enzymatic and chemical methods 443:490 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 3 45 theme chemical 475:482 arg1 methods 484:490 Different enzymatic and chemical methods 451:490 METHODS Different enzymatic and chemical methods 443:490 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 0 46 theme developmental 11:23 arg1 N-glycomes 34:43 developmental specific N-glycomes 11:43 developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum 11:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 1 47 theme nodule 118:123 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 47 theme nodule 118:123 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 6 48 theme fucosylated 1021:1031 arg1 glycans 1033:1039 fucosylated glycans 1021:1039 fucosylated glycans 1021:1039 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 3 49 theme enzymatic 461:469 arg1 methods 484:490 Different enzymatic and chemical methods 451:490 METHODS Different enzymatic and chemical methods 443:490 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 7 50 gly fucosylated 1110:1120 arg1 glycans 1138:1144 the fucosylated paucimannosidic glycans 1106:1144 the fucosylated paucimannosidic glycans 1106:1144 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 2 51 theme developmental 399:411 arg1 comparison 422:431 developmental glycomic comparison 399:431 developmental glycomic comparison 399:431 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 7 52 theme fucosylated 1110:1120 arg1 glycans 1138:1144 the fucosylated paucimannosidic glycans 1106:1144 the fucosylated paucimannosidic glycans 1106:1144 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 9 53 theme parasitic 1815:1823 arg1 nematode 1825:1832 a parasitic nematode 1813:1832 a parasitic nematode 1813:1832 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 2 54 theme obligate 343:350 arg1 organism 359:366 an obligate sexual organism 340:366 an obligate sexual organism 340:366 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 2 54 theme obligate 343:350 arg1 dentatum 328:335 the parasitic O. dentatum 311:335 the parasitic O. dentatum 311:335 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 5 55 theme phosphorylcholine-modified 768:793 arg1 structures 795:804 phosphorylcholine-modified structures 768:804 phosphorylcholine-modified structures 768:804 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 56 theme elegans 1255:1261 arg1 glycans 1241:1247 the tetrafucosylated glycans 1220:1247 the tetrafucosylated glycans of C. elegans 1220:1261 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 1 57 theme Caenorhabditis 233:246 arg1 elegans 248:254 the model organism Caenorhabditis elegans 214:254 the model organism Caenorhabditis elegans 214:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 5 58 theme male 811:814 arg1 worms 827:831 the male and female worms 807:831 the male and female worms 807:831 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 1 59 theme strongylid 160:169 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 59 theme strongylid 160:169 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 6 60 from females 1064:1070 arg1 pronounced 1050:1059 pronounced 1050:1059 pronounced 1050:1059 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 7 61 theme tetrafucosylated 1224:1239 arg1 glycans 1241:1247 the tetrafucosylated glycans 1220:1247 the tetrafucosylated glycans of C. elegans 1220:1261 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 7 62 theme other 1080:1084 arg1 hand 1086:1089 the other hand 1076:1089 the other hand 1076:1089 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 2 63 dep elegans 302:308 arg1 contrast 269:276 contrast 269:276 contrast 269:276 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 6 64 from pronounced 1050:1059 arg1 females 1064:1070 females 1064:1070 females 1064:1070 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 2 65 theme non-parasitic 285:297 arg1 elegans 302:308 the non-parasitic C. elegans 281:308 the non-parasitic C. elegans 281:308 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 2 66 theme O. 325:326 arg1 organism 359:366 an obligate sexual organism 340:366 an obligate sexual organism 340:366 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 2 66 theme O. 325:326 arg1 dentatum 328:335 the parasitic O. dentatum 311:335 the parasitic O. dentatum 311:335 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 8 67 theme sexual 1653:1658 arg1 differentiation 1660:1674 sexual differentiation 1653:1674 sexual differentiation 1653:1674 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 4 68 theme normal 650:655 arg1 phase 671:675 normal then reversed phase 650:675 normal then reversed phase 650:675 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 4 68 theme normal 650:655 arg1 2D-HPLC 641:647 2D-HPLC 641:647 2D-HPLC (normal then reversed phase) 641:676 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 2 69 theme parasitic 315:323 arg1 organism 359:366 an obligate sexual organism 340:366 an obligate sexual organism 340:366 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 2 69 theme parasitic 315:323 arg1 dentatum 328:335 the parasitic O. dentatum 311:335 the parasitic O. dentatum 311:335 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 6 70 theme methylated 936:945 arg1 structures 963:972 simple methylated paucimannosidic structures 929:972 simple methylated paucimannosidic structures 929:972 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 8 71 theme interspecies 1432:1443 arg1 tendency 1445:1452 The interspecies tendency 1428:1452 The interspecies tendency of class V nematodes 1428:1473 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 7 72 with identical 1150:1158 arg1 examples 1165:1172 examples 1165:1172 examples from other nematode species 1165:1200 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 4 73 theme MALDI-TOF 615:623 arg1 MS 625:626 MALDI-TOF MS 615:626 MALDI-TOF MS 615:626 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 9 74 dep methylated 1706:1715 arg1 Unusual 1698:1704 Unusual 1698:1704 Unusual 1698:1704 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 1 75 theme model 218:222 arg1 elegans 248:254 the model organism Caenorhabditis elegans 214:254 the model organism Caenorhabditis elegans 214:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 76 theme worm 125:128 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 76 theme worm 125:128 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 9 77 theme GENERAL 1677:1683 arg1 SIGNIFICANCE 1685:1696 GENERAL SIGNIFICANCE 1677:1696 GENERAL SIGNIFICANCE Unusual methylated 1677:1715 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 8 78 theme V 1463:1463 arg1 nematodes 1465:1473 class V nematodes 1457:1473 class V nematodes 1457:1473 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 27751954 4 79 theme fused 681:685 arg1 RP-HPLC 692:698 fused core RP-HPLC 681:698 fused core RP-HPLC 681:698 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 7 80 theme HexNAc2-4PC2-4 1338:1351 arg1 motifs 1353:1358 extended HexNAc2-4PC2-4 motifs 1329:1358 extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1329:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 7 81 with glycans 1316:1322 arg1 motifs 1353:1358 extended HexNAc2-4PC2-4 motifs 1329:1358 extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1329:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 5 82 theme core 857:860 arg1 N-glycans 874:882 core fucosylated N-glycans 857:882 core fucosylated N-glycans with up to three fucose residues 857:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 1 83 theme Oesophagostomum 130:144 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 83 theme Oesophagostomum 130:144 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 4 84 theme reversed 662:669 arg1 phase 671:675 normal then reversed phase 650:675 normal then reversed phase 650:675 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 4 84 theme reversed 662:669 arg1 2D-HPLC 641:647 2D-HPLC 641:647 2D-HPLC (normal then reversed phase) 641:676 Glycans were analysed by MALDI-TOF MS after either 2D-HPLC (normal then reversed phase) or fused core RP-HPLC. 27751954 7 85 theme nematode 1185:1192 arg1 species 1194:1200 other nematode species 1179:1200 other nematode species 1179:1200 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 5 86 theme N-glycans 874:882 arg1 range 848:852 a wide range 841:852 a wide range of core fucosylated N-glycans with up to three fucose residues 841:915 RESULTS Whereas the L3 N-glycome was simpler and more dominated by phosphorylcholine-modified structures, the male and female worms express a wide range of core fucosylated N-glycans with up to three fucose residues. 27751954 7 87 theme previous 1380:1387 arg1 studies 1389:1395 our previous studies 1376:1395 our previous studies on other nematodes 1376:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 9 88 theme phosphorylcholine-containing 1739:1766 arg1 N-glycans 1768:1776 phosphorylcholine-containing N-glycans 1739:1776 phosphorylcholine-containing N-glycans 1739:1776 GENERAL SIGNIFICANCE Unusual methylated, core fucosylated and phosphorylcholine-containing N-glycans vary between stages and genders in a parasitic nematode. 27751954 3 89 theme Different 451:459 arg1 methods 484:490 Different enzymatic and chemical methods 451:490 METHODS Different enzymatic and chemical methods 443:490 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 7 90 theme wide 1275:1278 arg1 range 1280:1284 a wide range 1273:1284 a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes 1273:1414 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 0 91 theme specific 25:32 arg1 N-glycomes 34:43 developmental specific N-glycomes 11:43 developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum 11:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 7 92 gly tetrafucosylated 1224:1239 arg1 glycans 1241:1247 the tetrafucosylated glycans 1220:1247 the tetrafucosylated glycans of C. elegans 1220:1261 On the other hand, while many of the fucosylated paucimannosidic glycans are identical with examples from other nematode species, but simpler than the tetrafucosylated glycans of C. elegans, there is a wide range of phosphorylcholine-modified glycans with extended HexNAc2-4PC2-4 motifs not observed in our previous studies on other nematodes. 27751954 1 93 theme porcine 110:116 arg1 dentatum 146:153 The porcine nodule worm Oesophagostomum dentatum 106:153 The porcine nodule worm Oesophagostomum dentatum 106:153 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 1 93 theme porcine 110:116 arg1 nematode 179:186 a strongylid class V nematode 158:186 a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans 158:254 BACKGROUND The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. 27751954 6 94 theme glycans 1033:1039 arg1 methylation 1006:1016 methylation 1006:1016 methylation of fucosylated glycans 1006:1039 Seemingly, simple methylated paucimannosidic structures can be considered 'male', while methylation of fucosylated glycans was more pronounced in females. 27751954 2 95 theme glycomic 413:420 arg1 comparison 422:431 developmental glycomic comparison 399:431 developmental glycomic comparison 399:431 However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. 27751954 0 96 theme porcine 52:58 arg1 dentatum 85:92 the porcine parasite Oesophagostomum dentatum 48:92 the porcine parasite Oesophagostomum dentatum 48:92 Gender and developmental specific N-glycomes of the porcine parasite Oesophagostomum dentatum. 27751954 3 97 dep METHODS 443:449 arg1 methods 484:490 Different enzymatic and chemical methods 451:490 METHODS Different enzymatic and chemical methods 443:490 METHODS Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae. 27751954 8 98 theme parasitic 1593:1601 arg1 nematode 1603:1610 a parasitic nematode 1591:1610 a parasitic nematode 1591:1610 CONCLUSION The interspecies tendency of class V nematodes to share most, but not all, N-glycans applies also to O. dentatum; furthermore, we establish, for the first time in a parasitic nematode, that glycomes vary upon development and sexual differentiation. 25129715 2 0 theme new 637:639 arg1 types 641:645 new types 637:645 new types of glycosidic linkages in all thermally treated mixtures 637:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 1 theme polymerization 557:570 arg1 degree 547:552 a degree 545:552 a degree of polymerization up to 18 residues 545:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 5 2 theme roasted 1144:1150 arg1 samples 1174:1180 roasted coffee polysaccharide samples 1144:1180 roasted coffee polysaccharide samples 1144:1180 These structural features were also found in roasted coffee polysaccharide samples. 25129715 1 3 theme → 281:281 arg1 -D-mannotriose 285:298 1 → 4)-D-mannotriose 279:298 1 → 4)-D-mannotriose 279:298 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 4 4 theme starting 1021:1028 arg1 mixtures 1030:1037 the starting mixtures 1017:1037 the starting mixtures 1017:1037 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 1 5 dep β- 276:277 arg1 α- 304:305 α-(1 → 5 304:311 α-(1 → 5 304:311 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 5 dep β- 276:277 arg1 -D-mannotriose 285:298 1 → 4)-D-mannotriose 279:298 1 → 4)-D-mannotriose 279:298 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 6 theme side 208:211 arg1 chains 213:218 side chains 208:218 side chains 208:218 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 4 7 dep arabinan 1053:1060 arg1 polysaccharides 1082:1096 chimeric polysaccharides 1073:1096 chimeric polysaccharides 1073:1096 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 1 8 theme dry 348:350 arg1 treatments 360:369 dry thermal treatments 348:369 dry thermal treatments 348:369 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 2 9 theme terminally 734:743 arg1 ribose 758:763 terminally and 5-linked ribose 734:763 terminally and 5-linked ribose 734:763 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 10 theme thermal 352:358 arg1 treatments 360:369 dry thermal treatments 348:369 dry thermal treatments 348:369 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 11 dep residues 581:588 arg1 up 572:573 up 572:573 up 572:573 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 5 12 theme coffee 1152:1157 arg1 samples 1174:1180 roasted coffee polysaccharide samples 1144:1180 roasted coffee polysaccharide samples 1144:1180 These structural features were also found in roasted coffee polysaccharide samples. 25129715 2 13 theme linkages 661:668 arg1 types 641:645 new types 637:645 new types of glycosidic linkages in all thermally treated mixtures 637:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 2 14 link 5-linked 749:756 arg1 ribose 758:763 terminally and 5-linked ribose 734:763 terminally and 5-linked ribose 734:763 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 5 15 theme polysaccharide 1159:1172 arg1 samples 1174:1180 roasted coffee polysaccharide samples 1144:1180 roasted coffee polysaccharide samples 1144:1180 These structural features were also found in roasted coffee polysaccharide samples. 25129715 1 16 with residues 531:538 arg1 degree 547:552 a degree 545:552 a degree of polymerization up to 18 residues 545:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 17 theme 200 374:376 arg1 °C. 378:380 °C. 378:380 °C. 378:380 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 18 theme arabinogalactans 223:238 arg1 occurrence 138:147 the possible occurrence 125:147 the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting 125:261 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 0 19 theme Transglycosylation 0:17 arg1 reactions 19:27 Transglycosylation reactions 0:27 Transglycosylation reactions between galactomannans and arabinogalactans 0:71 Transglycosylation reactions between galactomannans and arabinogalactans during dry thermal treatment. 25129715 4 20 from oligosaccharides 997:1012 arg1 mixtures 1030:1037 the starting mixtures 1017:1037 the starting mixtures 1017:1037 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 2 21 theme types 641:645 arg1 occurrence 720:729 the occurrence 716:729 the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization 716:809 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 2 21 theme types 641:645 arg1 occurrence 623:632 the occurrence 619:632 the occurrence of new types of glycosidic linkages in all thermally treated mixtures 619:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 2 22 theme glycosidic 650:659 arg1 linkages 661:668 glycosidic linkages 650:668 glycosidic linkages in all thermally treated mixtures 650:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 5 23 located found 1135:1139 arg1 samples 1174:1180 roasted coffee polysaccharide samples 1144:1180 roasted coffee polysaccharide samples 1144:1180 These structural features were also found in roasted coffee polysaccharide samples. 25129715 5 23 located found 1135:1139 arg2 features 1116:1123 These structural features 1099:1123 These structural features 1099:1123 These structural features were also found in roasted coffee polysaccharide samples. 25129715 4 24 theme reactions 951:959 arg1 occurrence 918:927 the occurrence 914:927 the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures 914:1037 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 1 25 theme pentose 512:518 arg1 residues 531:538 pentose and hexose residues 512:538 residues 531:538 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 2 26 theme treated 687:693 arg1 mixtures 695:702 all thermally treated mixtures 673:702 all thermally treated mixtures 673:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 27 theme coffee 247:252 arg1 roasting 254:261 coffee roasting 247:261 coffee roasting 247:261 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 4 28 theme chimeric 1073:1080 arg1 polysaccharides 1082:1096 chimeric polysaccharides 1073:1096 chimeric polysaccharides 1073:1096 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 4 29 theme transglycosylation 932:949 arg1 reactions 951:959 transglycosylation reactions 932:959 transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures 932:1037 These results support the occurrence of transglycosylation reactions promoted by roasting involving both oligosaccharides in the starting mixtures, resulting in arabinan and mannan chimeric polysaccharides. 25129715 1 30 theme °C. 378:380 arg1 analysis 455:462 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis 374:462 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 31 theme possible 129:136 arg1 occurrence 138:147 the possible occurrence 125:147 the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting 125:261 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 32 theme Matrix-assisted 382:396 arg1 spectrometry 431:442 Matrix-assisted laser desorption/ionization mass spectrometry 382:442 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 32 theme Matrix-assisted 382:396 arg1 MALDI-MS 445:452 MALDI-MS 445:452 MALDI-MS 445:452 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 2 33 theme arabinose 787:795 arg1 isomerization 797:809 arabinose isomerization 787:809 arabinose isomerization 787:809 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 34 theme laser 398:402 arg1 spectrometry 431:442 Matrix-assisted laser desorption/ionization mass spectrometry 382:442 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 34 theme laser 398:402 arg1 MALDI-MS 445:452 MALDI-MS 445:452 MALDI-MS 445:452 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 0 35 theme dry 80:82 arg1 treatment 92:100 dry thermal treatment 80:100 dry thermal treatment 80:100 Transglycosylation reactions between galactomannans and arabinogalactans during dry thermal treatment. 25129715 2 36 from types 641:645 arg1 mixtures 695:702 all thermally treated mixtures 673:702 all thermally treated mixtures 673:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 37 theme desorption/ionization 404:424 arg1 spectrometry 431:442 Matrix-assisted laser desorption/ionization mass spectrometry 382:442 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 37 theme desorption/ionization 404:424 arg1 MALDI-MS 445:452 MALDI-MS 445:452 MALDI-MS 445:452 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 38 theme hexose 524:529 arg1 residues 531:538 pentose and hexose residues 512:538 residues 531:538 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 2 39 from linkages 661:668 arg1 mixtures 695:702 all thermally treated mixtures 673:702 all thermally treated mixtures 673:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 40 theme mass 426:429 arg1 spectrometry 431:442 Matrix-assisted laser desorption/ionization mass spectrometry 382:442 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 40 theme mass 426:429 arg1 MALDI-MS 445:452 MALDI-MS 445:452 MALDI-MS 445:452 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 41 dep -D-mannotriose 285:298 arg1 4 283:283 4 283:283 4 283:283 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 42 theme -L-arabinotriose 313:328 arg1 mixtures 264:271 mixtures 264:271 mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose 264:328 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 2 43 theme Methylation 591:601 arg1 analysis 603:610 Methylation analysis 591:610 Methylation analysis 591:610 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 44 theme spectrometry 431:442 arg1 analysis 455:462 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis 374:462 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues 374:588 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 45 theme transglycosylation 152:169 arg1 reactions 171:179 transglycosylation reactions 152:179 transglycosylation reactions between galactomannans and side chains 152:218 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 5 46 theme structural 1105:1114 arg1 features 1116:1123 These structural features 1099:1123 These structural features 1099:1123 These structural features were also found in roasted coffee polysaccharide samples. 25129715 2 47 theme ribose 758:763 arg1 occurrence 720:729 the occurrence 716:729 the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization 716:809 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 2 47 theme ribose 758:763 arg1 occurrence 623:632 the occurrence 619:632 the occurrence of new types of glycosidic linkages in all thermally treated mixtures 619:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 1 48 theme reactions 171:179 arg1 occurrence 138:147 the possible occurrence 125:147 the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting 125:261 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 1 49 theme β- 276:277 arg1 -L-arabinotriose 313:328 β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose 276:328 β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose 276:328 Aiming to investigate the possible occurrence of transglycosylation reactions between galactomannans and side chains of arabinogalactans during coffee roasting, mixtures of β-(1 → 4)-D-mannotriose and α-(1 → 5)-L-arabinotriose were subjected to dry thermal treatments at 200 °C. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis allowed identifying polysaccharides composed by pentose and hexose residues with a degree of polymerization up to 18 residues. 25129715 0 50 dep reactions 19:27 arg1 treatment 92:100 dry thermal treatment 80:100 dry thermal treatment 80:100 Transglycosylation reactions between galactomannans and arabinogalactans during dry thermal treatment. 25129715 2 51 theme 5-linked 749:756 arg1 ribose 758:763 terminally and 5-linked ribose 734:763 terminally and 5-linked ribose 734:763 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 25129715 0 52 theme thermal 84:90 arg1 treatment 92:100 dry thermal treatment 80:100 dry thermal treatment 80:100 Transglycosylation reactions between galactomannans and arabinogalactans during dry thermal treatment. 25129715 2 53 from mixtures 695:702 arg1 types 641:645 new types 637:645 new types of glycosidic linkages in all thermally treated mixtures 637:702 Methylation analysis showed the occurrence of new types of glycosidic linkages in all thermally treated mixtures, as well as the occurrence of terminally and 5-linked ribose, possibly formed from arabinose isomerization. 28370937 0 0 theme Cells 84:88 arg1 N-Glycome 17:25 The Cell-Surface N-Glycome 0:25 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells 0:88 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 7 1 gly trisialylated 1000:1012 arg1 structures 1027:1036 trisialylated triantennary structures 1000:1036 trisialylated triantennary structures predominant 1000:1048 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 5 2 theme high-mannose 613:624 arg1 N-glycans 626:634 high-mannose N-glycans 613:634 high-mannose N-glycans 613:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 6 3 theme monosialylated 696:709 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 1 4 theme pluripotent 138:148 arg1 cells 155:159 pluripotent stem cells 138:159 pluripotent stem cells that offer a wide range of applications in regenerative medicine 138:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 4 theme pluripotent 138:148 arg1 cells 120:124 Human embryonic stem cells 99:124 Human embryonic stem cells (hESCs) 99:132 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 7 5 theme PHHs 910:913 arg1 N-glycosylation 891:905 The cell-surface N-glycosylation 874:905 The cell-surface N-glycosylation of PHHs 874:913 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 0 6 theme Hepatic 76:82 arg1 Cells 84:88 Differentiated Hepatic Cells 61:88 Differentiated Hepatic Cells 61:88 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 3 7 dep differentiated 342:355 arg1 followed 398:405 followed 398:405 followed by maturation into hepatocyte-like cells (HLCs) 398:453 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 7 8 theme bisialylated 971:982 arg1 biantennary 984:994 bisialylated biantennary 971:994 bisialylated biantennary 971:994 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 1 9 theme stem 150:153 arg1 cells 155:159 pluripotent stem cells 138:159 pluripotent stem cells that offer a wide range of applications in regenerative medicine 138:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 9 theme stem 150:153 arg1 cells 120:124 Human embryonic stem cells 99:124 Human embryonic stem cells (hESCs) 99:132 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 7 10 theme cell-surface 878:889 arg1 N-glycosylation 891:905 The cell-surface N-glycosylation 874:905 The cell-surface N-glycosylation of PHHs 874:913 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 4 11 theme human 537:541 arg1 PHHs 556:559 PHHs 556:559 PHHs 556:559 Their cell-surface N-glycome was profiled and also compared with that of primary human hepatocytes (PHHs). 28370937 4 11 theme human 537:541 arg1 hepatocytes 543:553 primary human hepatocytes 529:553 primary human hepatocytes (PHHs) 529:560 Their cell-surface N-glycome was profiled and also compared with that of primary human hepatocytes (PHHs). 28370937 7 12 theme trisialylated 1000:1012 arg1 structures 1027:1036 trisialylated triantennary structures 1000:1036 trisialylated triantennary structures predominant 1000:1048 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 7 13 theme triantennary 1014:1025 arg1 structures 1027:1036 trisialylated triantennary structures 1000:1036 trisialylated triantennary structures predominant 1000:1048 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 5 14 theme Undifferentiated 563:578 arg1 hESCs 580:584 Undifferentiated hESCs 563:584 Undifferentiated hESCs 563:584 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 8 15 theme first 1063:1067 arg1 report 1069:1074 the first report 1059:1074 the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation 1059:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 15 theme first 1063:1067 arg1 This 1051:1054 This 1051:1054 This 1051:1054 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 5 16 contain contained 586:594 arg2 N-glycans 626:634 high-mannose N-glycans 613:634 high-mannose N-glycans 613:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 5 16 contain contained 586:594 arg2 amounts 602:608 large amounts 596:608 large amounts of high-mannose N-glycans 596:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 5 16 contain contained 586:594 arg1 hESCs 580:584 Undifferentiated hESCs 563:584 Undifferentiated hESCs 563:584 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 8 17 theme cell 1083:1086 arg1 N-glycome 1096:1104 the cell surface N-glycome 1079:1104 the cell surface N-glycome 1079:1104 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 6 18 theme galactosylated 783:796 arg1 structures 798:807 fully galactosylated structures 777:807 fully galactosylated structures 777:807 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 19 theme asialylated 681:691 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 5 20 theme large 596:600 arg1 amounts 602:608 large amounts 596:608 large amounts of high-mannose N-glycans 596:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 5 20 theme large 596:600 arg1 N-glycans 626:634 high-mannose N-glycans 613:634 high-mannose N-glycans 613:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 0 21 theme Cell-Surface 4:15 arg1 N-Glycome 17:25 The Cell-Surface N-Glycome 0:25 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells 0:88 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 8 22 theme N-glycome 1096:1104 arg1 report 1069:1074 the first report 1059:1074 the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation 1059:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 22 theme N-glycome 1096:1104 arg1 This 1051:1054 This 1051:1054 This 1051:1054 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 3 23 theme definitive 362:371 arg1 cells 384:388 definitive endodermal cells 362:388 definitive endodermal cells (DECs) 362:395 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 3 23 theme definitive 362:371 arg1 DECs 391:394 DECs 391:394 DECs 391:394 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 8 24 theme surface 1088:1094 arg1 N-glycome 1096:1104 the cell surface N-glycome 1079:1104 the cell surface N-glycome 1079:1104 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 2 25 theme alternative 282:292 arg1 they 240:243 they 240:243 they 240:243 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 2 25 theme alternative 282:292 arg1 source 294:299 an appropriate alternative source 267:299 an appropriate alternative source of hepatocytes 267:314 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 8 26 theme embryoid 1170:1177 arg1 formation 1184:1192 embryoid body formation 1170:1192 embryoid body formation 1170:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 5 27 theme N-glycans 626:634 arg1 amounts 602:608 large amounts 596:608 large amounts of high-mannose N-glycans 596:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 5 27 theme N-glycans 626:634 arg1 N-glycans 626:634 high-mannose N-glycans 613:634 high-mannose N-glycans 613:634 Undifferentiated hESCs contained large amounts of high-mannose N-glycans. 28370937 8 28 from report 1069:1074 arg1 hESCs 1156:1160 hESCs 1156:1160 hESCs without embryoid body formation 1156:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 29 theme PHHs 1109:1112 arg1 report 1069:1074 the first report 1059:1074 the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation 1059:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 29 theme PHHs 1109:1112 arg1 This 1051:1054 This 1051:1054 This 1051:1054 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 2 30 theme appropriate 270:280 arg1 they 240:243 they 240:243 they 240:243 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 2 30 theme appropriate 270:280 arg1 source 294:299 an appropriate alternative source 267:299 an appropriate alternative source of hepatocytes 267:314 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 6 31 theme complex-type 650:661 arg1 N-glycans 663:671 complex-type N-glycans 650:671 complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans 650:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 31 theme complex-type 650:661 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 1 32 theme wide 174:177 arg1 range 179:183 a wide range 172:183 a wide range of applications in regenerative medicine 172:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 6 33 from contrast 640:647 arg1 dominant 755:762 dominant 755:762 dominant 755:762 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 4 34 theme primary 529:535 arg1 PHHs 556:559 PHHs 556:559 PHHs 556:559 Their cell-surface N-glycome was profiled and also compared with that of primary human hepatocytes (PHHs). 28370937 4 34 theme primary 529:535 arg1 hepatocytes 543:553 primary human hepatocytes 529:553 primary human hepatocytes (PHHs) 529:560 Their cell-surface N-glycome was profiled and also compared with that of primary human hepatocytes (PHHs). 28370937 1 35 theme embryonic 105:113 arg1 cells 155:159 pluripotent stem cells 138:159 pluripotent stem cells that offer a wide range of applications in regenerative medicine 138:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 35 theme embryonic 105:113 arg1 hESCs 127:131 hESCs 127:131 hESCs 127:131 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 35 theme embryonic 105:113 arg1 cells 120:124 Human embryonic stem cells 99:124 Human embryonic stem cells (hESCs) 99:132 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 6 36 from HLCs 767:770 arg1 dominant 755:762 dominant 755:762 dominant 755:762 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 0 37 theme Embryonic 36:44 arg1 Cells 51:55 Human Embryonic Stem Cells 30:55 Human Embryonic Stem Cells 30:55 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 0 38 theme Human 30:34 arg1 Cells 51:55 Human Embryonic Stem Cells 30:55 Human Embryonic Stem Cells 30:55 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 7 39 gly bisialylated 971:982 arg1 biantennary 984:994 bisialylated biantennary 971:994 bisialylated biantennary 971:994 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 7 40 gly N-glycosylation 891:905 arg1 PHHs 910:913 PHHs 910:913 PHHs 910:913 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 1 41 theme applications 188:199 arg1 range 179:183 a wide range 172:183 a wide range of applications in regenerative medicine 172:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 0 42 theme Cells 51:55 arg1 N-Glycome 17:25 The Cell-Surface N-Glycome 0:25 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells 0:88 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 1 43 from medicine 217:224 arg1 range 179:183 a wide range 172:183 a wide range of applications in regenerative medicine 172:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 6 44 theme undifferentiated 850:865 arg1 hESCs 867:871 undifferentiated hESCs 850:871 undifferentiated hESCs 850:871 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 1 45 from applications 188:199 arg1 medicine 217:224 regenerative medicine 204:224 regenerative medicine 204:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 46 theme stem 115:118 arg1 cells 155:159 pluripotent stem cells 138:159 pluripotent stem cells that offer a wide range of applications in regenerative medicine 138:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 46 theme stem 115:118 arg1 hESCs 127:131 hESCs 127:131 hESCs 127:131 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 46 theme stem 115:118 arg1 cells 120:124 Human embryonic stem cells 99:124 Human embryonic stem cells (hESCs) 99:132 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 0 47 theme Stem 46:49 arg1 Cells 51:55 Human Embryonic Stem Cells 30:55 Human Embryonic Stem Cells 30:55 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 6 48 gly asialylated 681:691 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 7 49 theme predominant 1038:1048 arg1 structures 1027:1036 trisialylated triantennary structures 1000:1036 trisialylated triantennary structures predominant 1000:1048 The cell-surface N-glycosylation of PHHs was more biologically processed than that of HLCs, with bisialylated biantennary and trisialylated triantennary structures predominant. 28370937 8 50 from hESCs 1156:1160 arg1 report 1069:1074 the first report 1059:1074 the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation 1059:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 50 from hESCs 1156:1160 arg1 This 1051:1054 This 1051:1054 This 1051:1054 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 51 theme generated 1141:1149 arg1 report 1069:1074 the first report 1059:1074 the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation 1059:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 8 51 theme generated 1141:1149 arg1 This 1051:1054 This 1051:1054 This 1051:1054 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 28370937 1 52 from range 179:183 arg1 medicine 217:224 regenerative medicine 204:224 regenerative medicine 204:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 2 53 theme hepatocytes 304:314 arg1 they 240:243 they 240:243 they 240:243 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 2 53 theme hepatocytes 304:314 arg1 source 294:299 an appropriate alternative source 267:299 an appropriate alternative source of hepatocytes 267:314 In addition, they have been proposed as an appropriate alternative source of hepatocytes. 28370937 1 54 theme Human 99:103 arg1 cells 155:159 pluripotent stem cells 138:159 pluripotent stem cells that offer a wide range of applications in regenerative medicine 138:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 54 theme Human 99:103 arg1 hESCs 127:131 hESCs 127:131 hESCs 127:131 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 1 54 theme Human 99:103 arg1 cells 120:124 Human embryonic stem cells 99:124 Human embryonic stem cells (hESCs) 99:132 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 6 55 theme triantennary 727:738 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 56 from dominant 755:762 arg1 contrast 640:647 contrast 640:647 contrast 640:647 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 56 from dominant 755:762 arg1 HLCs 767:770 HLCs 767:770 HLCs 767:770 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 1 57 theme regenerative 204:215 arg1 medicine 217:224 regenerative medicine 204:224 regenerative medicine 204:224 Human embryonic stem cells (hESCs) are pluripotent stem cells that offer a wide range of applications in regenerative medicine. 28370937 0 58 theme Differentiated 61:74 arg1 Cells 84:88 Differentiated Hepatic Cells 61:88 Differentiated Hepatic Cells 61:88 The Cell-Surface N-Glycome of Human Embryonic Stem Cells and Differentiated Hepatic Cells thereof. 28370937 3 59 theme hepatocyte-like 426:440 arg1 HLCs 449:452 HLCs 449:452 HLCs 449:452 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 3 59 theme hepatocyte-like 426:440 arg1 cells 442:446 hepatocyte-like cells 426:446 hepatocyte-like cells (HLCs) 426:453 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 6 60 gly monosialylated 696:709 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 3 61 theme endodermal 373:382 arg1 cells 384:388 definitive endodermal cells 362:388 definitive endodermal cells (DECs) 362:395 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 3 61 theme endodermal 373:382 arg1 DECs 391:394 DECs 391:394 DECs 391:394 In this work, hESCs were differentiated into definitive endodermal cells (DECs), followed by maturation into hepatocyte-like cells (HLCs). 28370937 4 62 theme cell-surface 462:473 arg1 N-glycome 475:483 Their cell-surface N-glycome 456:483 Their cell-surface N-glycome 456:483 Their cell-surface N-glycome was profiled and also compared with that of primary human hepatocytes (PHHs). 28370937 6 63 from abundant 833:840 arg1 hESCs 867:871 undifferentiated hESCs 850:871 undifferentiated hESCs 850:871 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 64 theme biantennary 711:721 arg1 N-glycans 740:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 asialylated or monosialylated biantennary and triantennary N-glycans 681:748 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 6 65 from hESCs 867:871 arg1 abundant 833:840 abundant 833:840 abundant 833:840 In contrast, complex-type N-glycans such as asialylated or monosialylated biantennary and triantennary N-glycans were dominant in HLCs, and fully galactosylated structures were significantly more abundant than in undifferentiated hESCs. 28370937 8 66 theme body 1179:1182 arg1 formation 1184:1192 embryoid body formation 1170:1192 embryoid body formation 1170:1192 This is the first report of the cell surface N-glycome of PHHs and of HLCs being directly generated from hESCs without embryoid body formation. 29085225 12 0 theme genes 1460:1464 arg1 Six 1447:1449 Six 1447:1449 Six 1447:1449 Six of these genes are involved in glycosylation-related processes. 29085225 12 0 theme genes 1460:1464 arg1 genes 1460:1464 these genes 1454:1464 these genes 1454:1464 Six of these genes are involved in glycosylation-related processes. 29085225 15 1 theme ontology 1715:1722 arg1 analysis 1724:1731 Gene ontology analysis 1710:1731 Gene ontology analysis 1710:1731 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 9 2 theme bacteria 1000:1007 arg1 isolation 982:990 the isolation 978:990 the isolation of live bacteria after 24 h incubation 978:1029 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 10 3 theme host 1239:1242 arg1 cell 1244:1247 the host cell 1235:1247 the host cell 1235:1247 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 3 4 theme host 432:435 arg1 cells 437:441 both infected and non-infected host cells 401:441 both infected and non-infected host cells 401:441 After infection RNA was isolated from both infected and non-infected host cells. 29085225 9 5 theme 24 1015:1016 arg1 h 1018:1018 h 1018:1018 h 1018:1018 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 2 6 theme Polarised 282:290 arg1 cells 305:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. 29085225 16 7 theme glycan 1885:1890 arg1 synthesis 1892:1900 glycan synthesis 1885:1900 glycan synthesis 1885:1900 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 8 8 dep structure 821:829 arg1 glycosyltransferases 832:851 glycosyltransferases 832:851 glycosyltransferases 832:851 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 8 8 dep structure 821:829 arg1 mucins 875:880 mucins 875:880 mucins 875:880 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 8 8 dep structure 821:829 arg1 lectins 866:872 lectins 866:872 lectins 866:872 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 8 8 dep structure 821:829 arg1 hydrolases 854:863 hydrolases 854:863 hydrolases 854:863 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 11 9 theme pylori 1370:1375 arg1 infection 1377:1385 H. pylori infection 1367:1385 H. pylori infection 1367:1385 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 13 10 theme mucin 1601:1605 arg1 MUC20 1607:1611 the mucin MUC20 1597:1611 the mucin MUC20 1597:1611 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 1 11 theme differential 199:210 arg1 expression 212:221 differential expression 199:221 differential expression on infection with Helicobacter pylori (H. pylori) 199:271 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 5 12 theme Affymetrix 598:607 arg1 arrays 609:614 Affymetrix arrays 598:614 Affymetrix arrays 598:614 RNA was isolated and hybridised to Affymetrix arrays. 29085225 7 13 theme ontology 746:753 arg1 categories 766:775 gene ontology functional categories 741:775 gene ontology functional categories 741:775 Genes were grouped into gene ontology functional categories. 29085225 1 14 dep H. 262:263 arg1 pylori 265:270 H. pylori 262:270 H. pylori 262:270 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 9 15 theme genes 1077:1081 arg1 amplification 1042:1054 PCR amplification 1038:1054 PCR amplification of bacteria-specific genes from the host cell RNA 1038:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 16 16 with infection 1839:1847 arg1 H. 1854:1855 H. 1854:1855 H. 1854:1855 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 9 17 theme bacteria-specific 1059:1075 arg1 genes 1077:1081 bacteria-specific genes 1059:1081 bacteria-specific genes from the host cell RNA 1059:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 8 18 theme glycan 814:819 arg1 structure 821:829 host glycan structure 809:829 host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) 809:881 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 1 19 from expression 212:221 arg1 infection 226:234 infection 226:234 infection with Helicobacter pylori (H. pylori) 226:271 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 3 20 attach isolated 387:394 arg2 RNA 379:381 RNA 379:381 RNA 379:381 After infection RNA was isolated from both infected and non-infected host cells. 29085225 3 20 attach isolated 387:394 arg1 cells 437:441 both infected and non-infected host cells 401:441 both infected and non-infected host cells 401:441 After infection RNA was isolated from both infected and non-infected host cells. 29085225 15 21 theme pylori 1776:1781 arg1 infection 1783:1791 H. pylori infection 1773:1791 H. pylori infection 1773:1791 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 1 22 theme HT29 150:153 arg1 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 1 22 theme HT29 150:153 arg1 line 171:174 the HT29 derivative cell line 146:174 the HT29 derivative cell line 146:174 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 9 23 theme host 1092:1095 arg1 RNA 1102:1104 the host cell RNA 1088:1104 the host cell RNA 1088:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 13 24 theme P 1584:1584 arg1 < 1586:1586 P < 0.05 1584:1591 P < 0.05 1584:1591 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 1 25 theme cell 166:169 arg1 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 1 25 theme cell 166:169 arg1 line 171:174 the HT29 derivative cell line 146:174 the HT29 derivative cell line 146:174 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 8 26 theme real-time 901:909 arg1 analysis 919:926 real-time qRT-PCR analysis 901:926 real-time qRT-PCR analysis 901:926 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 0 27 from expression 27:36 arg1 cells 54:58 HT29-MTX-E12 cells 41:58 HT29-MTX-E12 cells 41:58 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 29085225 15 28 theme previous 1753:1760 arg1 studies 1762:1768 previous studies 1753:1768 previous studies on H. pylori infection 1753:1791 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 9 29 from amplification 1042:1054 arg1 RNA 1102:1104 the host cell RNA 1088:1104 the host cell RNA 1088:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 6 30 theme data 640:643 arg1 Analysis 617:624 Analysis 617:624 Analysis of microarray data 617:643 Analysis of microarray data identified genes significantly differentially expressed upon infection. 29085225 10 31 theme adopted 1153:1159 arg1 conditions 1169:1178 the adopted culture conditions 1149:1178 the adopted culture conditions 1149:1178 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 16 32 theme Gene 1805:1808 arg1 data 1821:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 4 33 theme triplicate 494:503 arg1 samples 505:511 triplicate samples 494:511 triplicate samples for microarray analysis and for qRT-PCR analysis 494:560 Sufficient infections were carried out to provide triplicate samples for microarray analysis and for qRT-PCR analysis. 29085225 13 34 theme significant 1546:1556 arg1 1.8-fold 1574:1581 1.8-fold 1574:1581 1.8-fold 1574:1581 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 13 34 theme significant 1546:1556 arg1 downregulation 1558:1571 significant downregulation 1546:1571 significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20 1546:1611 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 0 35 theme Glycosylation-related 0:20 arg1 expression 27:36 Glycosylation-related gene expression 0:36 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 0:97 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 29085225 1 36 from genes 137:141 arg1 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 1 36 from genes 137:141 arg1 line 171:174 the HT29 derivative cell line 146:174 the HT29 derivative cell line 146:174 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 11 37 theme Microarray 1250:1259 arg1 analysis 1261:1268 Microarray analysis 1250:1268 Microarray analysis 1250:1268 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 15 38 with consistent 1737:1746 arg1 studies 1762:1768 previous studies 1753:1768 previous studies on H. pylori infection 1753:1791 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 1 39 theme glycosylation-related 115:135 arg1 genes 137:141 glycosylation-related genes 115:141 glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori) 115:271 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 0 40 theme HT29-MTX-E12 41:52 arg1 cells 54:58 HT29-MTX-E12 cells 41:58 HT29-MTX-E12 cells 41:58 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 29085225 10 41 theme adherent 1211:1218 arg1 layer 1226:1230 the adherent mucus layer 1207:1230 the adherent mucus layer of the host cell 1207:1247 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 16 42 theme glycan 1936:1941 arg1 structures 1943:1952 shorter and simpler glycan structures 1916:1952 shorter and simpler glycan structures 1916:1952 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 13 43 dep 1.8-fold 1574:1581 arg1 < 1586:1586 P < 0.05 1584:1591 P < 0.05 1584:1591 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 11 44 theme genes 1296:1300 arg1 total 1283:1287 a total 1281:1287 a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2 1281:1444 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 9 45 theme h 1018:1018 arg1 incubation 1020:1029 24 h incubation 1015:1029 24 h incubation 1015:1029 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 2 46 theme H. 339:340 arg1 strain 349:354 H. pylori strain 26695 339:360 H. pylori strain 26695 339:360 METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. 29085225 16 47 theme CONCLUSION 1794:1803 arg1 data 1821:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 6 48 theme microarray 629:638 arg1 data 640:643 microarray data 629:643 microarray data 629:643 Analysis of microarray data identified genes significantly differentially expressed upon infection. 29085225 11 49 theme fold 1401:1404 arg1 greater 1431:1437 greater 1431:1437 greater 1431:1437 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 11 49 theme fold 1401:1404 arg1 change 1406:1411 the fold change 1397:1411 the fold change in expression 1397:1425 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 9 50 theme host 950:953 arg1 cells 955:959 host cells 950:959 host cells 950:959 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 9 51 dep RESULTS 929:935 arg1 confirmed 965:973 confirmed 965:973 was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA 961:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 9 52 theme live 995:998 arg1 bacteria 1000:1007 live bacteria 995:1007 live bacteria 995:1007 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 14 53 dep downregulated 1659:1671 arg1 3.1-fold 1674:1681 3.1-fold 1674:1681 3.1-fold 1674:1681 REG4 was heavily expressed and significantly downregulated (3.1-fold, P < 0.05) upon infection. 29085225 10 54 theme cell 1244:1247 arg1 layer 1226:1230 the adherent mucus layer 1207:1230 the adherent mucus layer of the host cell 1207:1247 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 14 55 dep 3.1-fold 1674:1681 arg1 < 1686:1686 P < 0.05 1684:1691 P < 0.05 1684:1691 REG4 was heavily expressed and significantly downregulated (3.1-fold, P < 0.05) upon infection. 29085225 15 56 theme Gene 1710:1713 arg1 analysis 1724:1731 Gene ontology analysis 1710:1731 Gene ontology analysis 1710:1731 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 2 57 theme HT29-MTX-E12 292:303 arg1 cells 305:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. 29085225 11 58 theme H. 1367:1368 arg1 infection 1377:1385 H. pylori infection 1367:1385 H. pylori infection 1367:1385 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 16 59 from decrease 1873:1880 arg1 synthesis 1892:1900 glycan synthesis 1885:1900 glycan synthesis 1885:1900 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 15 60 from studies 1762:1768 arg1 infection 1783:1791 H. pylori infection 1773:1791 H. pylori infection 1773:1791 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 13 61 theme MUC20 1607:1611 arg1 1.8-fold 1574:1581 1.8-fold 1574:1581 1.8-fold 1574:1581 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 13 61 theme MUC20 1607:1611 arg1 downregulation 1558:1571 significant downregulation 1546:1571 significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20 1546:1611 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 16 62 theme shorter 1916:1922 arg1 structures 1943:1952 shorter and simpler glycan structures 1916:1952 shorter and simpler glycan structures 1916:1952 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 9 63 theme PCR 1038:1040 arg1 amplification 1042:1054 PCR amplification 1038:1054 PCR amplification of bacteria-specific genes from the host cell RNA 1038:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 8 64 theme Selected 778:785 arg1 genes 787:791 Selected genes 778:791 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) 778:881 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 7 65 theme functional 755:764 arg1 categories 766:775 gene ontology functional categories 741:775 gene ontology functional categories 741:775 Genes were grouped into gene ontology functional categories. 29085225 16 66 theme simpler 1928:1934 arg1 structures 1943:1952 shorter and simpler glycan structures 1916:1952 shorter and simpler glycan structures 1916:1952 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 7 67 theme gene 741:744 arg1 categories 766:775 gene ontology functional categories 741:775 gene ontology functional categories 741:775 Genes were grouped into gene ontology functional categories. 29085225 15 68 theme H. 1773:1774 arg1 infection 1783:1791 H. pylori infection 1773:1791 H. pylori infection 1773:1791 Gene ontology analysis was consistent with previous studies on H. pylori infection. 29085225 8 69 theme host 809:812 arg1 structure 821:829 host glycan structure 809:829 host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) 809:881 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 11 70 from change 1406:1411 arg1 expression 1416:1425 expression 1416:1425 expression 1416:1425 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 2 71 theme METHODS 274:280 arg1 cells 305:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells 274:309 METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. 29085225 4 72 theme Sufficient 444:453 arg1 infections 455:464 Sufficient infections 444:464 Sufficient infections 444:464 Sufficient infections were carried out to provide triplicate samples for microarray analysis and for qRT-PCR analysis. 29085225 0 73 theme Helicobacter 78:89 arg1 pylori 91:96 Helicobacter pylori 78:96 Helicobacter pylori 78:96 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 29085225 1 74 theme derivative 155:164 arg1 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 HT29-MTX-E12 177:188 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 1 74 theme derivative 155:164 arg1 line 171:174 the HT29 derivative cell line 146:174 the HT29 derivative cell line 146:174 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 10 75 dep H. 1107:1108 arg1 pylori 1110:1115 H. pylori 1107:1115 H. pylori 1107:1115 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 4 76 theme microarray 517:526 arg1 analysis 528:535 microarray analysis 517:535 microarray analysis 517:535 Sufficient infections were carried out to provide triplicate samples for microarray analysis and for qRT-PCR analysis. 29085225 9 77 theme cell 1097:1100 arg1 RNA 1102:1104 the host cell RNA 1088:1104 the host cell RNA 1088:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 14 78 theme P 1684:1684 arg1 < 1686:1686 P < 0.05 1684:1691 P < 0.05 1684:1691 REG4 was heavily expressed and significantly downregulated (3.1-fold, P < 0.05) upon infection. 29085225 8 79 theme qRT-PCR 911:917 arg1 analysis 919:926 real-time qRT-PCR analysis 901:926 real-time qRT-PCR analysis 901:926 Selected genes associated with host glycan structure (glycosyltransferases, hydrolases, lectins, mucins) were validated by real-time qRT-PCR analysis. 29085225 9 80 from RNA 1102:1104 arg1 amplification 1042:1054 PCR amplification 1038:1054 PCR amplification of bacteria-specific genes from the host cell RNA 1038:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 9 80 from RNA 1102:1104 arg1 genes 1077:1081 bacteria-specific genes 1059:1081 bacteria-specific genes from the host cell RNA 1059:1104 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 0 81 theme gene 22:25 arg1 expression 27:36 Glycosylation-related gene expression 0:36 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 0:97 Glycosylation-related gene expression in HT29-MTX-E12 cells upon infection by Helicobacter pylori. 29085225 11 82 theme P 1352:1352 arg1 < 1354:1354 P < 0.05 1352:1359 P < 0.05 1352:1359 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 1 83 with infection 226:234 arg1 H. 262:263 H. 262:263 H. 262:263 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 1 83 with infection 226:234 arg1 pylori 254:259 pylori 254:259 pylori 254:259 AIM To identify glycosylation-related genes in the HT29 derivative cell line, HT29-MTX-E12, showing differential expression on infection with Helicobacter pylori (H. pylori). 29085225 11 84 dep expressed 1341:1349 arg1 < 1354:1354 P < 0.05 1352:1359 P < 0.05 1352:1359 Microarray analysis identified a total of 276 genes that were significantly differentially expressed (P < 0.05) upon H. pylori infection and where the fold change in expression was greater than 2. 29085225 10 85 theme culture 1161:1167 arg1 conditions 1169:1178 the adopted culture conditions 1149:1178 the adopted culture conditions 1149:1178 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 29085225 3 86 theme non-infected 419:430 arg1 cells 437:441 both infected and non-infected host cells 401:441 both infected and non-infected host cells 401:441 After infection RNA was isolated from both infected and non-infected host cells. 29085225 4 87 theme qRT-PCR 545:551 arg1 analysis 553:560 qRT-PCR analysis 545:560 qRT-PCR analysis 545:560 Sufficient infections were carried out to provide triplicate samples for microarray analysis and for qRT-PCR analysis. 29085225 13 88 theme Real-time 1515:1523 arg1 qRT-PCR 1525:1531 Real-time qRT-PCR 1515:1531 Real-time qRT-PCR 1515:1531 Real-time qRT-PCR demonstrated significant downregulation (1.8-fold, P < 0.05) of the mucin MUC20. 29085225 16 89 theme expression 1810:1819 arg1 data 1821:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data 1794:1824 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 3 90 theme infected 406:413 arg1 cells 437:441 both infected and non-infected host cells 401:441 both infected and non-infected host cells 401:441 After infection RNA was isolated from both infected and non-infected host cells. 29085225 9 91 theme cells 955:959 arg1 Infection 937:945 Infection 937:945 Infection of host cells 937:959 RESULTS Infection of host cells was confirmed by the isolation of live bacteria after 24 h incubation and by PCR amplification of bacteria-specific genes from the host cell RNA. 29085225 2 92 theme pylori 342:347 arg1 strain 349:354 H. pylori strain 26695 339:360 H. pylori strain 26695 339:360 METHODS Polarised HT29-MTX-E12 cells were infected for 24 h with H. pylori strain 26695. 29085225 12 93 theme glycosylation-related 1482:1502 arg1 processes 1504:1512 glycosylation-related processes 1482:1512 glycosylation-related processes 1482:1512 Six of these genes are involved in glycosylation-related processes. 29085225 16 94 dep H. 1854:1855 arg1 pylori 1857:1862 H. pylori 1854:1862 H. pylori 1854:1862 CONCLUSION Gene expression data suggest that infection with H. pylori causes a decrease in glycan synthesis, resulting in shorter and simpler glycan structures. 29085225 10 95 theme mucus 1220:1224 arg1 layer 1226:1230 the adherent mucus layer 1207:1230 the adherent mucus layer of the host cell 1207:1247 H. pylori do not survive incubation under the adopted culture conditions unless they associate with the adherent mucus layer of the host cell. 28039139 9 0 theme glycoprotein 1744:1755 arg1 folding 1757:1763 S-layer glycoprotein folding 1736:1763 S-layer glycoprotein folding 1736:1763 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 12 1 theme archaeal 2366:2373 arg1 N-glycosylation 2375:2389 archaeal N-glycosylation 2366:2389 archaeal N-glycosylation 2366:2389 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 2 from Relying 2507:2513 arg1 strains 2525:2531 mutant strains 2518:2531 mutant strains defective in N-glycosylation 2518:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 5 3 from cells 1011:1015 arg1 vesicles 986:993 vesicles 986:993 vesicles from the mutant cells 986:1015 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 11 4 theme N-glycosylation 2199:2213 arg1 importance 2185:2194 the importance 2181:2194 the importance of N-glycosylation in eukaryotes and bacteria 2181:2240 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 12 5 theme species 2480:2486 arg1 Relying 2507:2513 the halophilic species Haloferax volcanii Relying 2465:2513 the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2465:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 6 6 theme mutant 1237:1242 arg1 strains 1244:1250 the parent and N-glycosylation mutant strains 1206:1250 strains 1244:1250 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 1 7 theme proteins 173:180 arg1 residues 154:161 select Asn residues 143:161 select Asn residues of target proteins 143:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 7 theme proteins 173:180 arg1 proteins 173:180 target proteins 166:180 target proteins 166:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 10 8 from changes 1821:1827 arg1 surroundings 1836:1847 its surroundings 1832:1847 its surroundings 1832:1847 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 4 9 used used 555:558 arg2 microscopy 540:549 Cryo-electron microscopy 526:549 Cryo-electron microscopy 526:549 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 10 link N-linked 731:738 arg1 pentasaccharide 740:754 the N-linked pentasaccharide 727:754 the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells 727:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 12 11 theme compromised 2590:2600 arg1 N-glycosylation 2602:2616 compromised N-glycosylation 2590:2616 compromised N-glycosylation 2590:2616 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 10 12 theme sugars 2098:2103 arg1 attachment 2084:2093 the covalent attachment 2071:2093 the covalent attachment of sugars to select asparagine residues of target proteins 2071:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 12 theme sugars 2098:2103 arg1 N-glycosylation 2046:2060 N-glycosylation 2046:2060 N-glycosylation 2046:2060 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 12 13 from N-glycosylation 2546:2560 arg1 defective 2533:2541 defective 2533:2541 defective 2533:2541 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 1 14 theme posttranslational 206:222 arg1 modification 224:235 an almost universal posttranslational modification 186:235 an almost universal posttranslational modification in archaea 186:246 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 14 theme posttranslational 206:222 arg1 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 12 15 from strains 2525:2531 arg1 cells 2456:2460 cells 2456:2460 cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2456:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 4 16 theme surface 771:777 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 16 theme surface 771:777 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 12 17 from defective 2533:2541 arg1 N-glycosylation 2546:2560 N-glycosylation 2546:2560 N-glycosylation 2546:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 5 18 theme strain 972:977 arg1 cells 979:983 parent strain cells 965:983 parent strain cells 965:983 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 12 19 theme S-layer 2771:2777 arg1 component 2802:2810 the sole component 2793:2810 the sole component of the S-layer 2793:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 19 theme S-layer 2771:2777 arg1 glycoprotein 2779:2790 the S-layer glycoprotein 2767:2790 the S-layer glycoprotein 2767:2790 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 20 gly N-glycosylation 2602:2616 arg1 S-layer 2627:2633 S-layer integrity 2627:2643 S-layer integrity 2627:2643 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 20 gly N-glycosylation 2602:2616 arg1 component 2802:2810 the sole component 2793:2810 the sole component of the S-layer 2793:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 20 gly N-glycosylation 2602:2616 arg1 protein 2685:2691 a secreted reporter protein 2665:2691 a secreted reporter protein 2665:2691 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 20 gly N-glycosylation 2602:2616 arg1 glycoprotein 2779:2790 the S-layer glycoprotein 2767:2790 the S-layer glycoprotein 2767:2790 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 6 21 theme N-glycosylation-related 1072:1094 arg1 effects 1096:1102 such N-glycosylation-related effects 1067:1102 such N-glycosylation-related effects on S-layer assembly 1067:1122 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 4 22 theme N-linked 731:738 arg1 pentasaccharide 740:754 the N-linked pentasaccharide 727:754 the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells 727:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 10 23 theme select 2108:2113 arg1 residues 2126:2133 select asparagine residues 2108:2133 select asparagine residues of target proteins 2108:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 23 theme select 2108:2113 arg1 proteins 2145:2152 target proteins 2138:2152 target proteins 2138:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 1 24 theme Asn 150:152 arg1 residues 154:161 select Asn residues 143:161 select Asn residues of target proteins 143:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 24 theme Asn 150:152 arg1 proteins 173:180 target proteins 166:180 target proteins 166:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 8 25 theme pentasaccharide 1564:1578 arg1 sugar 1580:1584 the final pentasaccharide sugar 1554:1584 the final pentasaccharide sugar 1554:1584 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 12 26 gly glycoprotein 2779:2790 arg1 component 2802:2810 the sole component 2793:2810 the sole component of the S-layer 2793:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 26 gly glycoprotein 2779:2790 arg1 glycoprotein 2779:2790 the S-layer glycoprotein 2767:2790 the S-layer glycoprotein 2767:2790 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 4 27 theme Cryo-electron 526:538 arg1 microscopy 540:549 Cryo-electron microscopy 526:549 Cryo-electron microscopy 526:549 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 6 28 from effects 1096:1102 arg1 assembly 1115:1122 S-layer assembly 1107:1122 S-layer assembly 1107:1122 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 12 29 theme growth 2721:2726 arg1 medium 2728:2733 the growth medium 2717:2733 the growth medium 2717:2733 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 4 30 theme sole 813:816 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 30 theme sole 813:816 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 1 31 theme covalent 112:119 arg1 linkage 121:127 the covalent linkage 108:127 the covalent linkage of glycans to select Asn residues of target proteins 108:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 31 theme covalent 112:119 arg1 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 10 32 theme target 2138:2143 arg1 proteins 2145:2152 target proteins 2138:2152 target proteins 2138:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 8 33 theme distinct 1589:1596 arg1 conformation 1619:1630 a distinct S-layer glycoprotein conformation 1587:1630 a distinct S-layer glycoprotein conformation 1587:1630 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 8 34 theme susceptibility 1440:1453 arg1 assessment 1405:1414 an assessment 1402:1414 an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants 1402:1487 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 7 35 theme S-layer 1265:1271 arg1 N-glycosylation 1286:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 8 36 from assessment 1405:1414 arg1 mutants 1481:1487 the mutants 1477:1487 the mutants 1477:1487 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 8 37 theme glycoprotein 1606:1617 arg1 conformation 1619:1630 a distinct S-layer glycoprotein conformation 1587:1630 a distinct S-layer glycoprotein conformation 1587:1630 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 8 38 theme added 1458:1462 arg1 proteases 1464:1472 added proteases 1458:1472 added proteases 1458:1472 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 3 39 theme N-glycosylation 423:437 arg1 impact 403:408 the impact 399:408 the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii 399:508 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 4 40 theme right-side-out 571:584 arg1 vesicles 595:602 right-side-out membrane vesicles 571:602 right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells 571:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 8 41 gly glycoprotein 1427:1438 arg1 glycoprotein 1427:1438 S-layer glycoprotein susceptibility 1419:1453 S-layer glycoprotein susceptibility to added proteases 1419:1472 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 5 42 theme mutant 1004:1009 arg1 cells 1011:1015 the mutant cells 1000:1015 the mutant cells 1000:1015 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 1 43 theme select 143:148 arg1 residues 154:161 select Asn residues 143:161 select Asn residues of target proteins 143:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 43 theme select 143:148 arg1 proteins 173:180 target proteins 166:180 target proteins 166:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 12 44 theme such 2563:2566 arg1 efforts 2568:2574 such efforts 2563:2574 such efforts 2563:2574 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 0 45 theme Proper 33:38 arg1 volcanii 50:57 Proper Haloferax volcanii 33:57 Proper Haloferax volcanii S-Layer Stability 33:75 N-Glycosylation Is Important for Proper Haloferax volcanii S-Layer Stability and Function. 28039139 11 46 from importance 2185:2194 arg1 eukaryotes 2218:2227 eukaryotes 2218:2227 eukaryotes 2218:2227 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 11 46 from importance 2185:2194 arg1 bacteria 2233:2240 bacteria 2233:2240 bacteria 2233:2240 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 7 47 theme growth 1378:1383 arg1 medium 1385:1390 the growth medium 1374:1390 the growth medium 1374:1390 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 11 48 theme posttranslational 2273:2289 arg1 modification 2291:2302 this posttranslational modification 2268:2302 this posttranslational modification 2268:2302 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 12 49 theme mutant 2518:2523 arg1 strains 2525:2531 mutant strains 2518:2531 mutant strains defective in N-glycosylation 2518:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 3 50 theme volcanii 501:508 arg1 physiology 460:469 physiology 460:469 physiology 460:469 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 3 50 theme volcanii 501:508 arg1 structure 446:454 structure 446:454 structure 446:454 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 0 51 theme volcanii 50:57 arg1 Stability 67:75 Proper Haloferax volcanii S-Layer Stability 33:75 Proper Haloferax volcanii S-Layer Stability 33:75 N-Glycosylation Is Important for Proper Haloferax volcanii S-Layer Stability and Function. 28039139 3 52 dep structure 446:454 arg1 the 442:444 the 442:444 the 442:444 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 12 53 theme reporter 2676:2683 arg1 protein 2685:2691 a secreted reporter protein 2665:2691 a secreted reporter protein 2665:2691 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 54 theme defective 2533:2541 arg1 strains 2525:2531 mutant strains 2518:2531 mutant strains defective in N-glycosylation 2518:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 4 55 theme S-layer 835:841 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 55 theme S-layer 835:841 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 10 56 attach attachment 2084:2093 arg2 sugars 2098:2103 sugars 2098:2103 sugars 2098:2103 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 56 attach attachment 2084:2093 arg1 residues 2126:2133 select asparagine residues 2108:2133 select asparagine residues of target proteins 2108:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 56 attach attachment 2084:2093 arg1 proteins 2145:2152 target proteins 2138:2152 target proteins 2138:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 3 57 theme haloarchaeon 478:489 arg1 volcanii 501:508 the haloarchaeon Haloferax volcanii 474:508 the haloarchaeon Haloferax volcanii 474:508 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 9 58 gly glycoprotein 1744:1755 arg1 glycoprotein 1744:1755 S-layer glycoprotein folding 1736:1763 S-layer glycoprotein folding 1736:1763 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 7 59 theme S-layer 1357:1363 arg1 transfer 1323:1330 impaired transfer 1314:1330 impaired transfer of the reporter past the S-layer 1314:1363 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 4 60 theme parent 629:634 arg1 strain 636:641 a parent strain 627:641 a parent strain 627:641 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 61 theme H. 855:856 arg1 cells 867:871 H. volcanii cells 855:871 H. volcanii cells 855:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 13 62 theme volcanii 2867:2874 arg1 cells 2876:2880 H. volcanii cells 2864:2880 H. volcanii cells 2864:2880 Thus, by modifying N-glycosylation, H. volcanii cells can change how they interact with their surroundings. 28039139 9 63 theme S-layer 1736:1742 arg1 folding 1757:1763 S-layer glycoprotein folding 1736:1763 S-layer glycoprotein folding 1736:1763 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 7 64 theme past 1348:1351 arg1 S-layer 1357:1363 the reporter past the S-layer 1335:1363 the reporter past the S-layer 1335:1363 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 12 65 theme Relying 2507:2513 arg1 cells 2456:2460 cells 2456:2460 cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2456:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 2 66 theme archaeal 351:358 arg1 N-glycosylation 360:374 archaeal N-glycosylation 351:374 archaeal N-glycosylation 351:374 However, whereas roles for N-glycosylation have been defined in eukarya and bacteria, the function of archaeal N-glycosylation remains unclear. 28039139 9 67 gly N-glycosylation 1707:1721 arg1 glycoprotein 1744:1755 S-layer glycoprotein folding 1736:1763 S-layer glycoprotein folding 1736:1763 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 9 67 gly N-glycosylation 1707:1721 arg1 S-layer 1736:1742 S-layer glycoprotein folding 1736:1763 S-layer glycoprotein folding 1736:1763 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 12 68 theme S-layer 2627:2633 arg1 integrity 2635:2643 S-layer integrity 2627:2643 S-layer integrity 2627:2643 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 69 theme halophilic 2469:2478 arg1 species 2480:2486 the halophilic species 2465:2486 the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2465:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 11 70 from role 2260:2263 arg1 archaea 2307:2313 archaea 2307:2313 archaea 2307:2313 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 1 71 theme target 166:171 arg1 proteins 173:180 target proteins 166:180 target proteins 166:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 6 72 theme N-glycosylation 1221:1235 arg1 strains 1244:1250 the parent and N-glycosylation mutant strains 1206:1250 strains 1244:1250 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 10 73 theme S-layer 1895:1901 arg1 architecture 1903:1914 S-layer architecture 1895:1914 S-layer architecture 1895:1914 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 5 74 theme entire 924:929 arg1 surface 931:937 the entire surface 920:937 the entire surface of vesicles prepared from parent strain cells 920:983 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 9 75 theme Perturbed 1697:1705 arg1 N-glycosylation 1707:1721 Perturbed N-glycosylation 1697:1721 Perturbed N-glycosylation 1697:1721 Perturbed N-glycosylation thus affects S-layer glycoprotein folding. 28039139 5 76 theme repeating 894:902 arg1 S-layer 904:910 a regularly repeating S-layer 882:910 a regularly repeating S-layer 882:910 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 12 77 theme S-layer 2819:2825 arg1 component 2802:2810 the sole component 2793:2810 the sole component of the S-layer 2793:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 77 theme S-layer 2819:2825 arg1 glycoprotein 2779:2790 the S-layer glycoprotein 2767:2790 the S-layer glycoprotein 2767:2790 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 10 78 dep function.IMPORTANCE 1920:1938 arg1 held 1945:1948 held 1945:1948 function.IMPORTANCE Long held to be a process unique to eukaryotes 1920:1985 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 5 79 theme parent 965:970 arg1 cells 979:983 parent strain cells 965:983 parent strain cells 965:983 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 12 80 theme surface 2420:2426 arg1 layer 2428:2432 the surface layer 2416:2432 the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2416:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 80 theme surface 2420:2426 arg1 S-layer 2435:2441 S-layer 2435:2441 S-layer 2435:2441 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 81 theme glycoprotein 2779:2790 arg1 conformation 2751:2762 the conformation 2747:2762 the conformation of the S-layer glycoprotein, the sole component of the S-layer 2747:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 81 theme glycoprotein 2779:2790 arg1 integrity 2635:2643 S-layer integrity 2627:2643 S-layer integrity 2627:2643 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 81 theme glycoprotein 2779:2790 arg1 transfer 2653:2660 the transfer 2649:2660 the transfer of a secreted reporter protein across the S-layer into the growth medium 2649:2733 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 10 82 theme covalent 2075:2082 arg1 attachment 2084:2093 the covalent attachment 2071:2093 the covalent attachment of sugars to select asparagine residues of target proteins 2071:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 82 theme covalent 2075:2082 arg1 N-glycosylation 2046:2060 N-glycosylation 2046:2060 N-glycosylation 2046:2060 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 1 83 from modification 224:235 arg1 archaea 240:246 archaea 240:246 archaea 240:246 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 84 theme glycans 132:138 arg1 linkage 121:127 the covalent linkage 108:127 the covalent linkage of glycans to select Asn residues of target proteins 108:180 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 84 theme glycans 132:138 arg1 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 12 85 theme sole 2797:2800 arg1 component 2802:2810 the sole component 2793:2810 the sole component of the S-layer 2793:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 85 theme sole 2797:2800 arg1 glycoprotein 2779:2790 the S-layer glycoprotein 2767:2790 the S-layer glycoprotein 2767:2790 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 8 86 theme final 1558:1562 arg1 sugar 1580:1584 the final pentasaccharide sugar 1554:1584 the final pentasaccharide sugar 1554:1584 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 8 87 theme S-layer 1419:1425 arg1 susceptibility 1440:1453 S-layer glycoprotein susceptibility 1419:1453 S-layer glycoprotein susceptibility to added proteases 1419:1472 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 4 88 theme layer 779:783 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 88 theme layer 779:783 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 10 89 theme asparagine 2115:2124 arg1 residues 2126:2133 select asparagine residues 2108:2133 select asparagine residues of target proteins 2108:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 89 theme asparagine 2115:2124 arg1 proteins 2145:2152 target proteins 2138:2152 target proteins 2138:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 7 90 theme impaired 1314:1321 arg1 transfer 1323:1330 impaired transfer 1314:1330 impaired transfer of the reporter past the S-layer 1314:1363 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 5 91 theme vesicles 942:949 arg1 surface 931:937 the entire surface 920:937 the entire surface of vesicles prepared from parent strain cells 920:983 Whereas a regularly repeating S-layer covered the entire surface of vesicles prepared from parent strain cells, vesicles from the mutant cells were only partially covered. 28039139 6 92 theme such 1067:1070 arg1 effects 1096:1102 such N-glycosylation-related effects 1067:1102 such N-glycosylation-related effects on S-layer assembly 1067:1122 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 12 93 theme volcanii 2498:2505 arg1 Relying 2507:2513 the halophilic species Haloferax volcanii Relying 2465:2513 the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2465:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 3 94 from impact 403:408 arg1 physiology 460:469 physiology 460:469 physiology 460:469 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 3 94 from impact 403:408 arg1 structure 446:454 structure 446:454 structure 446:454 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 8 95 gly glycoprotein 1606:1617 arg1 glycoprotein 1606:1617 a distinct S-layer glycoprotein conformation 1587:1630 a distinct S-layer glycoprotein conformation 1587:1630 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 10 96 theme proteins 2145:2152 arg1 residues 2126:2133 select asparagine residues 2108:2133 select asparagine residues of target proteins 2108:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 10 96 theme proteins 2145:2152 arg1 proteins 2145:2152 target proteins 2138:2152 target proteins 2138:2152 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 7 97 theme glycoprotein 1273:1284 arg1 N-glycosylation 1286:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 8 98 theme S-layer 1598:1604 arg1 conformation 1619:1630 a distinct S-layer glycoprotein conformation 1587:1630 a distinct S-layer glycoprotein conformation 1587:1630 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 7 99 gly glycoprotein 1273:1284 arg1 glycoprotein 1273:1284 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 6 100 theme reporter 1172:1179 arg1 protein 1181:1187 a reporter protein 1170:1187 a reporter protein 1170:1187 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 4 101 gly glycoprotein 795:806 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 101 gly glycoprotein 795:806 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 11 102 from information 2166:2176 arg1 importance 2185:2194 the importance 2181:2194 the importance of N-glycosylation in eukaryotes and bacteria 2181:2240 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 12 103 theme N-glycosylation 2375:2389 arg1 purpose 2355:2361 the purpose 2351:2361 the purpose of archaeal N-glycosylation 2351:2389 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 8 104 theme glycoprotein 1427:1438 arg1 susceptibility 1440:1453 S-layer glycoprotein susceptibility 1419:1453 S-layer glycoprotein susceptibility to added proteases 1419:1472 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 7 105 theme Compromised 1253:1263 arg1 N-glycosylation 1286:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation 1253:1300 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 12 106 from cells 2456:2460 arg1 strains 2525:2531 mutant strains 2518:2531 mutant strains defective in N-glycosylation 2518:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 6 107 theme cell 1138:1141 arg1 function 1143:1150 cell function 1138:1150 cell function 1138:1150 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 4 108 theme membrane 586:593 arg1 vesicles 595:602 right-side-out membrane vesicles 571:602 right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells 571:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 109 theme S-layer 786:792 arg1 glycoprotein 795:806 the surface layer (S-layer) glycoprotein 767:806 the surface layer (S-layer) glycoprotein 767:806 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 4 109 theme S-layer 786:792 arg1 component 818:826 the sole component 809:826 the sole component of the S-layer surrounding H. volcanii cells 809:871 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 12 110 theme Haloferax 2488:2496 arg1 Relying 2507:2513 the halophilic species Haloferax volcanii Relying 2465:2513 the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation 2465:2560 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 0 111 theme Haloferax 40:48 arg1 volcanii 50:57 Proper Haloferax volcanii 33:57 Proper Haloferax volcanii S-Layer Stability 33:75 N-Glycosylation Is Important for Proper Haloferax volcanii S-Layer Stability and Function. 28039139 1 112 theme universal 196:204 arg1 modification 224:235 an almost universal posttranslational modification 186:235 an almost universal posttranslational modification in archaea 186:246 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 1 112 theme universal 196:204 arg1 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation 91:105 N-Glycosylation, the covalent linkage of glycans to select Asn residues of target proteins, is an almost universal posttranslational modification in archaea. 28039139 6 113 theme S-layer 1107:1113 arg1 assembly 1115:1122 S-layer assembly 1107:1122 S-layer assembly 1107:1122 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 3 114 theme perturbed 413:421 arg1 N-glycosylation 423:437 perturbed N-glycosylation 413:437 perturbed N-glycosylation 413:437 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 8 115 theme N-terminal 1657:1666 arg1 region 1668:1673 the N-terminal region 1653:1673 the N-terminal region 1653:1673 Finally, an assessment of S-layer glycoprotein susceptibility to added proteases in the mutants revealed that in cells lacking AglD, which is involved in adding the final pentasaccharide sugar, a distinct S-layer glycoprotein conformation was assumed in which the N-terminal region was readily degraded. 28039139 10 116 theme unique 1966:1971 arg1 process 1958:1964 a process 1956:1964 a process unique to eukaryotes 1956:1985 These findings suggest that H. volcanii could adapt to changes in its surroundings by modulating N-glycosylation so as to affect S-layer architecture and function.IMPORTANCE Long held to be a process unique to eukaryotes, it is now accepted that bacteria and archaea also perform N-glycosylation, namely, the covalent attachment of sugars to select asparagine residues of target proteins. 28039139 0 117 theme S-Layer 59:65 arg1 Stability 67:75 Proper Haloferax volcanii S-Layer Stability 33:75 Proper Haloferax volcanii S-Layer Stability 33:75 N-Glycosylation Is Important for Proper Haloferax volcanii S-Layer Stability and Function. 28039139 11 118 theme modification 2291:2302 arg1 role 2260:2263 the role 2256:2263 the role of this posttranslational modification in archaea 2256:2313 Yet, while information on the importance of N-glycosylation in eukaryotes and bacteria is available, the role of this posttranslational modification in archaea remains unclear. 28039139 6 119 theme protein 1181:1187 arg1 secretion 1157:1165 the secretion 1153:1165 the secretion of a reporter protein 1153:1187 To determine whether such N-glycosylation-related effects on S-layer assembly also affected cell function, the secretion of a reporter protein was addressed in the parent and N-glycosylation mutant strains. 28039139 12 120 theme secreted 2667:2674 arg1 protein 2685:2691 a secreted reporter protein 2665:2691 a secreted reporter protein 2665:2691 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 4 121 theme strain 636:641 arg1 cells 618:622 cells 618:622 cells of a parent strain 618:641 Cryo-electron microscopy was used to examine right-side-out membrane vesicles prepared from cells of a parent strain and from strains lacking genes encoding glycosyltransferases involved in assembling the N-linked pentasaccharide decorating the surface layer (S-layer) glycoprotein, the sole component of the S-layer surrounding H. volcanii cells. 28039139 3 122 theme Haloferax 491:499 arg1 volcanii 501:508 the haloarchaeon Haloferax volcanii 474:508 the haloarchaeon Haloferax volcanii 474:508 Here, the impact of perturbed N-glycosylation on the structure and physiology of the haloarchaeon Haloferax volcanii was considered. 28039139 13 123 theme H. 2864:2865 arg1 cells 2876:2880 H. volcanii cells 2864:2880 H. volcanii cells 2864:2880 Thus, by modifying N-glycosylation, H. volcanii cells can change how they interact with their surroundings. 28039139 7 124 theme reporter 1339:1346 arg1 S-layer 1357:1363 the reporter past the S-layer 1335:1363 the reporter past the S-layer 1335:1363 Compromised S-layer glycoprotein N-glycosylation resulted in impaired transfer of the reporter past the S-layer and into the growth medium. 28039139 12 125 theme protein 2685:2691 arg1 conformation 2751:2762 the conformation 2747:2762 the conformation of the S-layer glycoprotein, the sole component of the S-layer 2747:2825 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 125 theme protein 2685:2691 arg1 integrity 2635:2643 S-layer integrity 2627:2643 S-layer integrity 2627:2643 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 12 125 theme protein 2685:2691 arg1 transfer 2653:2660 the transfer 2649:2660 the transfer of a secreted reporter protein across the S-layer into the growth medium 2649:2733 Here, insight into the purpose of archaeal N-glycosylation was gained by addressing the surface layer (S-layer) surrounding cells of the halophilic species Haloferax volcanii Relying on mutant strains defective in N-glycosylation, such efforts revealed that compromised N-glycosylation affected S-layer integrity and the transfer of a secreted reporter protein across the S-layer into the growth medium, as well as the conformation of the S-layer glycoprotein, the sole component of the S-layer. 28039139 2 126 theme N-glycosylation 360:374 arg1 function 339:346 the function 335:346 the function of archaeal N-glycosylation 335:374 However, whereas roles for N-glycosylation have been defined in eukarya and bacteria, the function of archaeal N-glycosylation remains unclear. 25482815 3 0 theme glycan 673:678 arg1 residues 680:687 glycan residues 673:687 glycan residues 673:687 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 4 1 theme N1141 830:834 arg1 flagellin 836:844 N1141 flagellin 830:844 N1141 flagellin 830:844 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 6 2 theme immature 1046:1053 arg1 flagellin 1071:1079 The immature glycan-attached flagellin 1042:1079 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, 1042:1120 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 5 3 from modification 1002:1013 arg1 flagellin 1031:1039 K1 flagellin 1028:1039 K1 flagellin 1028:1039 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 0 4 from moiety 7:12 arg1 K1 48:49 Acidovorax avenae K1 30:49 Acidovorax avenae K1 30:49 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 3 5 from K1 735:736 arg1 flagellins 709:718 the flagellins 705:718 the flagellins from N1141 and K1 705:736 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 6 theme Mass 601:604 arg1 analyses 619:626 Mass spectrometry analyses 601:626 Mass spectrometry analyses 601:626 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 2 7 theme avenae 554:559 arg1 flagellin 505:513 flagellin 505:513 flagellin from the rice virulent K1 strain of A. avenae 505:559 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 4 8 theme epitope 898:904 arg1 recognition 906:916 epitope recognition 898:916 epitope recognition 898:916 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 2 9 theme A. 551:552 arg1 avenae 554:559 A. avenae 551:559 A. avenae 551:559 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 10 from strain 541:546 arg1 flagellin 505:513 flagellin 505:513 flagellin from the rice virulent K1 strain of A. avenae 505:559 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 1 11 theme bacterial 245:253 arg1 flagellum 255:263 the bacterial flagellum 241:263 the bacterial flagellum 241:263 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 6 12 theme H2O2 1130:1133 arg1 generation 1135:1144 H2O2 generation 1130:1144 H2O2 generation 1130:1144 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 1 13 theme pathogen-associated 155:173 arg1 patterns 185:192 pathogen-associated molecular patterns 155:192 pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, 155:264 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 13 theme pathogen-associated 155:173 arg1 flagellin 210:218 flagellin 210:218 flagellin 210:218 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 13 theme pathogen-associated 155:173 arg1 PAMPs 195:199 PAMPs 195:199 PAMPs 195:199 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 2 14 theme H2O2 482:485 arg1 generation 487:496 H2O2 generation 482:496 H2O2 generation 482:496 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 1 15 theme PAMP-triggered 334:347 arg1 Recognition 140:150 Abstract Recognition 131:150 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, 131:264 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 15 theme PAMP-triggered 334:347 arg1 PTI 359:361 PTI 359:361 PTI 359:361 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 15 theme PAMP-triggered 334:347 arg1 immunity 349:356 PAMP-triggered immunity 334:356 PAMP-triggered immunity (PTI) 334:362 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 2 16 theme immune 583:588 arg1 responses 590:598 these immune responses 577:598 these immune responses 577:598 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 17 theme gram-negative 400:412 arg1 bacterium 430:438 gram-negative phytopathogenic bacterium 400:438 gram-negative phytopathogenic bacterium 400:438 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 17 theme gram-negative 400:412 arg1 avenae 452:457 Acidovorax avenae 441:457 Acidovorax avenae 441:457 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 5 18 from genes 947:951 arg1 operon 968:973 K1 flagella operon 956:973 K1 flagella operon 956:973 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 1 19 theme molecular 175:183 arg1 patterns 185:192 pathogen-associated molecular patterns 155:192 pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, 155:264 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 19 theme molecular 175:183 arg1 flagellin 210:218 flagellin 210:218 flagellin 210:218 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 19 theme molecular 175:183 arg1 PAMPs 195:199 PAMPs 195:199 PAMPs 195:199 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 2 20 theme phytopathogenic 414:428 arg1 bacterium 430:438 gram-negative phytopathogenic bacterium 400:438 gram-negative phytopathogenic bacterium 400:438 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 20 theme phytopathogenic 414:428 arg1 avenae 452:457 Acidovorax avenae 441:457 Acidovorax avenae 441:457 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 7 21 from recognition 1346:1356 arg1 rice 1361:1364 rice 1361:1364 rice 1361:1364 The data indicate that the immature glycan of flagellin from KΔ3FG cannot prevent the epitope recognition in rice. 25482815 1 22 theme patterns 185:192 arg1 Recognition 140:150 Abstract Recognition 131:150 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, 131:264 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 22 theme patterns 185:192 arg1 immunity 349:356 PAMP-triggered immunity 334:356 PAMP-triggered immunity (PTI) 334:362 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 0 23 theme Glycan 0:5 arg1 moiety 7:12 Glycan moiety 0:12 Glycan moiety of flagellin in Acidovorax avenae K1 0:49 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 1 24 theme flagellum 255:263 arg1 flagellin 210:218 flagellin 210:218 flagellin 210:218 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 24 theme flagellum 255:263 arg1 component 228:236 a main component 221:236 a main component of the bacterial flagellum 221:263 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 3 25 from N1141 725:729 arg1 flagellins 709:718 the flagellins 705:718 the flagellins from N1141 and K1 705:736 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 26 theme total 642:646 arg1 1,600-Da 648:655 total 1,600-Da 642:655 total 1,600-Da 642:655 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 7 27 from KΔ3FG 1313:1317 arg1 glycan 1288:1293 the immature glycan 1275:1293 the immature glycan of flagellin from KΔ3FG 1275:1317 The data indicate that the immature glycan of flagellin from KΔ3FG cannot prevent the epitope recognition in rice. 25482815 0 28 theme flagellin 17:25 arg1 moiety 7:12 Glycan moiety 0:12 Glycan moiety of flagellin in Acidovorax avenae K1 0:49 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 5 29 theme K1 956:957 arg1 operon 968:973 K1 flagella operon 956:973 K1 flagella operon 956:973 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 2 30 theme rice 524:527 arg1 strain 541:546 the rice virulent K1 strain 520:546 the rice virulent K1 strain 520:546 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 4 31 from glycan 867:872 arg1 flagellin 880:888 K1 flagellin 877:888 K1 flagellin 877:888 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 6 32 theme mature-type 1185:1195 arg1 flagellin 1197:1205 the K1 mature-type flagellin 1178:1205 the K1 mature-type flagellin 1178:1205 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 2 33 theme N1141 384:388 arg1 strain 390:395 The rice avirulent N1141 strain 365:395 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, 365:458 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 6 34 from mutant 1107:1112 arg1 flagellin 1071:1079 The immature glycan-attached flagellin 1042:1079 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, 1042:1120 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 3 35 theme spectrometry 606:617 arg1 analyses 619:626 Mass spectrometry analyses 601:626 Mass spectrometry analyses 601:626 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 6 36 theme rice 1158:1161 arg1 cells 1163:1167 cultured rice cells 1149:1167 cultured rice cells 1149:1167 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 2 37 theme avirulent 374:382 arg1 strain 390:395 The rice avirulent N1141 strain 365:395 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, 365:458 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 0 38 theme Acidovorax 30:39 arg1 K1 48:49 Acidovorax avenae K1 30:49 Acidovorax avenae K1 30:49 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 5 39 theme flagella 959:966 arg1 operon 968:973 K1 flagella operon 956:973 K1 flagella operon 956:973 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 0 40 theme responses 120:128 arg1 induction 100:108 the induction 96:108 the induction of immune responses 96:128 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 5 41 theme K1 1028:1029 arg1 flagellin 1031:1039 K1 flagellin 1028:1039 K1 flagellin 1028:1039 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 5 42 theme structural 991:1000 arg1 modification 1002:1013 structural modification 991:1013 structural modification of glycan in K1 flagellin 991:1039 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 6 43 theme cultured 1149:1156 arg1 cells 1163:1167 cultured rice cells 1149:1167 cultured rice cells 1149:1167 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 2 44 theme rice 369:372 arg1 strain 390:395 The rice avirulent N1141 strain 365:395 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, 365:458 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 45 theme Acidovorax 441:450 arg1 bacterium 430:438 gram-negative phytopathogenic bacterium 400:438 gram-negative phytopathogenic bacterium 400:438 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 2 45 theme Acidovorax 441:450 arg1 avenae 452:457 Acidovorax avenae 441:457 Acidovorax avenae 441:457 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 1 46 theme first 282:286 arg1 layer 288:292 the first layer 278:292 the first layer of plant immunity 278:310 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 0 47 theme immune 113:118 arg1 responses 120:128 immune responses 113:128 immune responses 113:128 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 6 48 theme K1 1182:1183 arg1 flagellin 1197:1205 the K1 mature-type flagellin 1178:1205 the K1 mature-type flagellin 1178:1205 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 4 49 gly deglycosylated 755:768 arg1 flagellin 773:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 3 50 located present 694:700 arg2 1,600-Da 648:655 total 1,600-Da 642:655 total 1,600-Da 642:655 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 50 located present 694:700 arg1 flagellins 709:718 the flagellins 705:718 the flagellins from N1141 and K1 705:736 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 50 located present 694:700 arg2 2,150-Da 661:668 2,150-Da 661:668 2,150-Da 661:668 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 51 theme residues 680:687 arg1 2,150-Da 661:668 2,150-Da 661:668 2,150-Da 661:668 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 51 theme residues 680:687 arg1 1,600-Da 648:655 total 1,600-Da 642:655 total 1,600-Da 642:655 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 6 52 theme deletion 1098:1105 arg1 KΔ3FG 1115:1119 KΔ3FG 1115:1119 KΔ3FG 1115:1119 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 6 52 theme deletion 1098:1105 arg1 mutant 1107:1112 three genes deletion mutant 1086:1112 three genes deletion mutant 1086:1112 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 0 53 theme avenae 41:46 arg1 K1 48:49 Acidovorax avenae K1 30:49 Acidovorax avenae K1 30:49 Glycan moiety of flagellin in Acidovorax avenae K1 prevents the recognition by rice that causes the induction of immune responses. 25482815 4 54 theme K1 877:878 arg1 flagellin 880:888 K1 flagellin 877:888 K1 flagellin 877:888 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 2 55 theme bacterium 430:438 arg1 strain 390:395 The rice avirulent N1141 strain 365:395 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, 365:458 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 4 56 theme immune 791:796 arg1 responses 798:806 immune responses 791:806 immune responses 791:806 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 7 57 theme immature 1279:1286 arg1 glycan 1288:1293 the immature glycan 1275:1293 the immature glycan of flagellin from KΔ3FG 1275:1317 The data indicate that the immature glycan of flagellin from KΔ3FG cannot prevent the epitope recognition in rice. 25482815 6 58 theme genes 1092:1096 arg1 KΔ3FG 1115:1119 KΔ3FG 1115:1119 KΔ3FG 1115:1119 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 6 58 theme genes 1092:1096 arg1 mutant 1107:1112 three genes deletion mutant 1086:1112 three genes deletion mutant 1086:1112 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 1 59 theme plant 297:301 arg1 immunity 303:310 plant immunity 297:310 plant immunity 297:310 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 2 60 theme K1 538:539 arg1 strain 541:546 the rice virulent K1 strain 520:546 the rice virulent K1 strain 520:546 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 7 61 theme flagellin 1298:1306 arg1 glycan 1288:1293 the immature glycan 1275:1293 the immature glycan of flagellin from KΔ3FG 1275:1317 The data indicate that the immature glycan of flagellin from KΔ3FG cannot prevent the epitope recognition in rice. 25482815 7 62 theme epitope 1338:1344 arg1 recognition 1346:1356 the epitope recognition 1334:1356 the epitope recognition in rice 1334:1364 The data indicate that the immature glycan of flagellin from KΔ3FG cannot prevent the epitope recognition in rice. 25482815 2 63 theme virulent 529:536 arg1 strain 541:546 the rice virulent K1 strain 520:546 the rice virulent K1 strain 520:546 The rice avirulent N1141 strain of gram-negative phytopathogenic bacterium, Acidovorax avenae, induces PTI including H2O2 generation, while flagellin from the rice virulent K1 strain of A. avenae does not induce these immune responses. 25482815 6 64 theme glycan-attached 1055:1069 arg1 flagellin 1071:1079 The immature glycan-attached flagellin 1042:1079 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, 1042:1120 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 5 65 theme glycan 1018:1023 arg1 modification 1002:1013 structural modification 991:1013 structural modification of glycan in K1 flagellin 991:1039 We identified three genes in K1 flagella operon, which regulate structural modification of glycan in K1 flagellin. 25482815 4 66 theme K1 770:771 arg1 flagellin 773:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 6 67 from increase 1234:1241 arg1 H2O2 1246:1249 H2O2 1246:1249 H2O2 1246:1249 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 3 68 attach present 694:700 arg2 1,600-Da 648:655 total 1,600-Da 642:655 total 1,600-Da 642:655 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 68 attach present 694:700 arg1 flagellins 709:718 the flagellins 705:718 the flagellins from N1141 and K1 705:736 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 3 68 attach present 694:700 arg2 2,150-Da 661:668 2,150-Da 661:668 2,150-Da 661:668 Mass spectrometry analyses revealed that total 1,600-Da and 2,150-Da of glycan residues were present on the flagellins from N1141 and K1, respectively. 25482815 1 69 theme immunity 303:310 arg1 layer 288:292 the first layer 278:292 the first layer of plant immunity 278:310 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 4 70 theme deglycosylated 755:768 arg1 flagellin 773:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin 753:781 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 4 71 theme same 815:818 arg1 manner 820:825 the same manner 811:825 the same manner as N1141 flagellin 811:844 A deglycosylated K1 flagellin induced immune responses in the same manner as N1141 flagellin, suggesting that the glycan in K1 flagellin prevent epitope recognition in rice. 25482815 1 72 theme Abstract 131:138 arg1 Recognition 140:150 Abstract Recognition 131:150 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, 131:264 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 72 theme Abstract 131:138 arg1 immunity 349:356 PAMP-triggered immunity 334:356 PAMP-triggered immunity (PTI) 334:362 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 6 73 theme detectable 1223:1232 arg1 increase 1234:1241 a detectable increase 1221:1241 a detectable increase in H2O2 1221:1249 The immature glycan-attached flagellin from three genes deletion mutant, KΔ3FG, induced H2O2 generation in cultured rice cells, whereas the K1 mature-type flagellin did not cause a detectable increase in H2O2. 25482815 1 74 theme main 223:226 arg1 flagellin 210:218 flagellin 210:218 flagellin 210:218 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 25482815 1 74 theme main 223:226 arg1 component 228:236 a main component 221:236 a main component of the bacterial flagellum 221:263 Abstract Recognition of pathogen-associated molecular patterns (PAMPs) such as flagellin, a main component of the bacterial flagellum, constitutes the first layer of plant immunity and is referred to as PAMP-triggered immunity (PTI). 29162910 1 0 theme major 171:175 arg1 bottleneck 177:186 a major bottleneck 169:186 a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards 169:310 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 0 1 theme oligosaccharide 75:89 arg1 precursors 91:100 microbial oligosaccharide precursors 65:100 microbial oligosaccharide precursors 65:100 A library of chemically defined human N-glycans synthesized from microbial oligosaccharide precursors. 29162910 5 2 theme high-throughput 1485:1499 arg1 screening 1521:1529 high-throughput, fluorescence-based screening 1485:1529 high-throughput, fluorescence-based screening of glycan-binding proteins 1485:1556 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 3 theme hybrid 1331:1336 arg1 N-glycans 1355:1363 hybrid and complex-type N-glycans 1331:1363 N-glycans 1355:1363 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 3 theme hybrid 1331:1336 arg1 structures 1402:1411 asymmetric multi-antennary structures 1375:1411 asymmetric multi-antennary structures 1375:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 4 4 theme biosynthetic 891:902 arg1 precursors 904:913 these biosynthetic precursors 885:913 these biosynthetic precursors 885:913 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 1 5 theme affinity 271:278 arg1 resins 280:285 affinity resins 271:285 affinity resins 271:285 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 4 6 theme key 1136:1138 arg1 N-acetylglucosaminyltransferases 1140:1171 key N-acetylglucosaminyltransferases 1136:1171 key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1136:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 4 7 theme structural 1027:1036 arg1 remodeling 1038:1047 structural remodeling 1027:1047 structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1027:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 3 8 theme direct 569:574 arg1 isolation 576:584 direct isolation 569:584 direct isolation 569:584 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 4 9 gly deglycosylation 866:880 arg1 precursors 904:913 these biosynthetic precursors 885:913 these biosynthetic precursors 885:913 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 2 10 theme biological/enzymatic 342:361 arg1 synthesis 363:371 a combined biological/enzymatic synthesis 331:371 a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans 331:507 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 2 10 theme biological/enzymatic 342:361 arg1 capable 381:387 capable 381:387 capable 381:387 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 4 11 theme simplified 984:993 arg1 scheme 1008:1013 a greatly simplified purification scheme 974:1013 a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 974:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 2 12 theme microbially-derived 415:433 arg1 oligosaccharides 445:460 microbially-derived precursor oligosaccharides 415:460 microbially-derived precursor oligosaccharides 415:460 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 3 13 attach derived 693:699 arg2 precursors 682:691 precursors 682:691 precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli 682:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 13 attach derived 693:699 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 13 attach derived 693:699 arg1 sources 716:722 renewable sources 706:722 renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli 706:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 13 attach derived 693:699 arg1 oligosaccharides 800:815 lipid-linked oligosaccharides 787:815 lipid-linked oligosaccharides from glycoengineered Escherichia coli 787:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 5 14 theme preparative 1305:1315 arg1 quantities 1317:1326 preparative quantities 1305:1326 preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures 1305:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 15 dep high-throughput 1485:1499 arg1 fluorescence-based 1502:1519 fluorescence-based 1502:1519 fluorescence-based 1502:1519 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 4 16 theme precursors 904:913 arg1 deglycosylation 866:880 deglycosylation 866:880 deglycosylation of these biosynthetic precursors 866:913 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 6 17 theme combined 1601:1608 arg1 strategy 1620:1627 our combined synthesis strategy 1597:1627 our combined synthesis strategy 1597:1627 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 6 17 theme combined 1601:1608 arg1 route 1653:1657 a new, user-friendly route 1632:1657 a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling 1632:1786 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 5 18 theme complex-type 1342:1353 arg1 N-glycans 1355:1363 hybrid and complex-type N-glycans 1331:1363 N-glycans 1355:1363 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 18 theme complex-type 1342:1353 arg1 structures 1402:1411 asymmetric multi-antennary structures 1375:1411 asymmetric multi-antennary structures 1375:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 19 theme asymmetric 1375:1384 arg1 structures 1402:1411 asymmetric multi-antennary structures 1375:1411 asymmetric multi-antennary structures 1375:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 6 20 theme human 1692:1696 arg1 glycans 1698:1704 chemically defined human glycans 1673:1704 chemically defined human glycans 1673:1704 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 4 21 from remodeling 1222:1231 arg1 pathway 1275:1281 the mammalian glycosylation pathway 1247:1281 the mammalian glycosylation pathway 1247:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 3 22 theme wild-type 734:742 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 5 23 theme proteins 1549:1556 arg1 screening 1521:1529 high-throughput, fluorescence-based screening 1485:1529 high-throughput, fluorescence-based screening of glycan-binding proteins 1485:1556 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 2 24 theme human-type 488:497 arg1 N-glycans 499:507 structurally uniform human-type N-glycans 467:507 structurally uniform human-type N-glycans 467:507 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 5 25 theme multi-antennary 1386:1400 arg1 structures 1402:1411 asymmetric multi-antennary structures 1375:1411 asymmetric multi-antennary structures 1375:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 4 26 theme produced 1096:1103 arg1 glycosyltransferases 1105:1124 commercially available and recombinantly produced glycosyltransferases 1055:1124 commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1055:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 4 26 theme produced 1096:1103 arg1 N-acetylglucosaminyltransferases 1140:1171 key N-acetylglucosaminyltransferases 1136:1171 key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1136:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 6 27 dep new 1634:1636 arg1 user-friendly 1639:1651 user-friendly 1639:1651 user-friendly 1639:1651 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 1 28 theme molecular 209:217 arg1 tools 219:223 molecular tools 209:223 molecular tools 209:223 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 1 28 theme molecular 209:217 arg1 standards 302:310 reference standards 292:310 reference standards 292:310 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 1 28 theme molecular 209:217 arg1 microarrays 258:268 glycan microarrays 251:268 glycan microarrays 251:268 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 1 28 theme molecular 209:217 arg1 resins 280:285 affinity resins 271:285 affinity resins 271:285 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 2 29 theme uniform 480:486 arg1 N-glycans 499:507 structurally uniform human-type N-glycans 467:507 structurally uniform human-type N-glycans 467:507 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 1 30 theme reference 292:300 arg1 standards 302:310 reference standards 292:310 reference standards 292:310 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 3 31 theme renewable 706:714 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 31 theme renewable 706:714 arg1 sources 716:722 renewable sources 706:722 renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli 706:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 31 theme renewable 706:714 arg1 oligosaccharides 800:815 lipid-linked oligosaccharides 787:815 lipid-linked oligosaccharides from glycoengineered Escherichia coli 787:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 1 32 theme homogenous 116:125 arg1 glycans 127:133 homogenous glycans 116:133 homogenous glycans 116:133 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 3 33 link lipid-linked 787:798 arg1 oligosaccharides 800:815 lipid-linked oligosaccharides 787:815 lipid-linked oligosaccharides from glycoengineered Escherichia coli 787:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 1 34 theme tools 219:223 arg1 production 195:204 the production 191:204 the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards 191:310 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 0 35 theme human 32:36 arg1 N-glycans 38:46 chemically defined human N-glycans 13:46 chemically defined human N-glycans 13:46 A library of chemically defined human N-glycans synthesized from microbial oligosaccharide precursors. 29162910 4 36 theme microbial 930:938 arg1 oligosaccharides 940:955 the resulting microbial oligosaccharides 916:955 the resulting microbial oligosaccharides 916:955 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 5 37 theme glycan-binding 1534:1547 arg1 proteins 1549:1556 glycan-binding proteins 1534:1556 glycan-binding proteins 1534:1556 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 0 38 theme defined 24:30 arg1 N-glycans 38:46 chemically defined human N-glycans 13:46 chemically defined human N-glycans 13:46 A library of chemically defined human N-glycans synthesized from microbial oligosaccharide precursors. 29162910 2 39 link microbially-derived 415:433 arg1 oligosaccharides 445:460 microbially-derived precursor oligosaccharides 415:460 microbially-derived precursor oligosaccharides 415:460 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 6 40 theme enzymatic 1767:1775 arg1 remodeling 1777:1786 enzymatic remodeling 1767:1786 enzymatic remodeling 1767:1786 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 2 41 theme precursor 435:443 arg1 oligosaccharides 445:460 microbially-derived precursor oligosaccharides 415:460 microbially-derived precursor oligosaccharides 415:460 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 4 42 theme early 1216:1220 arg1 remodeling 1222:1231 early remodeling 1216:1231 early remodeling of glycans in the mammalian glycosylation pathway 1216:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 4 43 dep GnTII 1186:1190 arg1 e.g. 1174:1177 e.g. 1174:1177 e.g. 1174:1177 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 4 44 theme purification 995:1006 arg1 scheme 1008:1013 a greatly simplified purification scheme 974:1013 a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 974:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 1 45 theme glycans 127:133 arg1 Synthesis 103:111 Synthesis 103:111 Synthesis of homogenous glycans in quantitative yields 103:156 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 0 46 theme N-glycans 38:46 arg1 library 2:8 A library 0:8 A library of chemically defined human N-glycans 0:46 A library of chemically defined human N-glycans synthesized from microbial oligosaccharide precursors. 29162910 5 47 theme N-glycans 1355:1363 arg1 quantities 1317:1326 preparative quantities 1305:1326 preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures 1305:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 5 48 used used 1445:1448 arg2 quantities 1317:1326 preparative quantities 1305:1326 preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures 1305:1411 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 2 49 theme combined 333:340 arg1 synthesis 363:371 a combined biological/enzymatic synthesis 331:371 a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans 331:507 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 2 49 theme combined 333:340 arg1 capable 381:387 capable 381:387 capable 381:387 Here, we describe a combined biological/enzymatic synthesis that is capable of efficiently converting microbially-derived precursor oligosaccharides into structurally uniform human-type N-glycans. 29162910 4 50 theme glycosylation 1261:1273 arg1 pathway 1275:1281 the mammalian glycosylation pathway 1247:1281 the mammalian glycosylation pathway 1247:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 3 51 gly glycoproteins 769:781 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 6 52 link biosynthetically-derived 1726:1749 arg1 precursors 1751:1760 biosynthetically-derived precursors 1726:1760 biosynthetically-derived precursors 1726:1760 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 3 53 theme cerevisiae 758:767 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 4 54 theme resulting 920:928 arg1 oligosaccharides 940:955 the resulting microbial oligosaccharides 916:955 the resulting microbial oligosaccharides 916:955 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 1 55 theme quantitative 138:149 arg1 yields 151:156 quantitative yields 138:156 quantitative yields 138:156 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 4 56 theme mammalian 1251:1259 arg1 pathway 1275:1281 the mammalian glycosylation pathway 1247:1281 the mammalian glycosylation pathway 1247:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 6 57 theme defined 1684:1690 arg1 glycans 1698:1704 chemically defined human glycans 1673:1704 chemically defined human glycans 1673:1704 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 4 58 theme available 1068:1076 arg1 glycosyltransferases 1105:1124 commercially available and recombinantly produced glycosyltransferases 1055:1124 commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1055:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 4 58 theme available 1068:1076 arg1 N-acetylglucosaminyltransferases 1140:1171 key N-acetylglucosaminyltransferases 1136:1171 key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway 1136:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 3 59 theme chemical 547:554 arg1 synthesis 556:564 chemical synthesis 547:564 chemical synthesis 547:564 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 6 60 theme synthesis 1610:1618 arg1 strategy 1620:1627 our combined synthesis strategy 1597:1627 our combined synthesis strategy 1597:1627 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 6 60 theme synthesis 1610:1618 arg1 route 1653:1657 a new, user-friendly route 1632:1657 a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling 1632:1786 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 0 61 theme microbial 65:73 arg1 precursors 91:100 microbial oligosaccharide precursors 65:100 microbial oligosaccharide precursors 65:100 A library of chemically defined human N-glycans synthesized from microbial oligosaccharide precursors. 29162910 3 62 theme starting 517:524 arg1 material 526:533 starting material 517:533 starting material 517:533 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 6 63 theme biosynthetically-derived 1726:1749 arg1 precursors 1751:1760 biosynthetically-derived precursors 1726:1760 biosynthetically-derived precursors 1726:1760 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 5 64 theme glycan 1463:1468 arg1 microarray 1470:1479 a glycan microarray 1461:1479 a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins 1461:1556 Using this approach, preparative quantities of hybrid and complex-type N-glycans including asymmetric multi-antennary structures were generated and subsequently used to develop a glycan microarray for high-throughput, fluorescence-based screening of glycan-binding proteins. 29162910 1 65 from Synthesis 103:111 arg1 yields 151:156 quantitative yields 138:156 quantitative yields 138:156 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 29162910 6 66 theme new 1634:1636 arg1 strategy 1620:1627 our combined synthesis strategy 1597:1627 our combined synthesis strategy 1597:1627 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 6 66 theme new 1634:1636 arg1 route 1653:1657 a new, user-friendly route 1632:1657 a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling 1632:1786 Taken together, these results confirm our combined synthesis strategy as a new, user-friendly route for supplying chemically defined human glycans simply by combining biosynthetically-derived precursors with enzymatic remodeling. 29162910 3 67 theme lipid-linked 787:798 arg1 oligosaccharides 800:815 lipid-linked oligosaccharides 787:815 lipid-linked oligosaccharides from glycoengineered Escherichia coli 787:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 68 from coli 850:853 arg1 glycoproteins 769:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 wild-type Saccharomyces cerevisiae glycoproteins 734:781 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 68 from coli 850:853 arg1 oligosaccharides 800:815 lipid-linked oligosaccharides 787:815 lipid-linked oligosaccharides from glycoengineered Escherichia coli 787:853 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 3 69 theme natural 591:597 arg1 sources 599:605 natural sources 591:605 natural sources 591:605 Unlike starting material obtained by chemical synthesis or direct isolation from natural sources, which can be time consuming and costly to generate, our approach involves precursors derived from renewable sources including wild-type Saccharomyces cerevisiae glycoproteins and lipid-linked oligosaccharides from glycoengineered Escherichia coli. 29162910 4 70 theme glycans 1236:1242 arg1 remodeling 1222:1231 early remodeling 1216:1231 early remodeling of glycans in the mammalian glycosylation pathway 1216:1281 Following deglycosylation of these biosynthetic precursors, the resulting microbial oligosaccharides are subjected to a greatly simplified purification scheme followed by structural remodeling using commercially available and recombinantly produced glycosyltransferases including key N-acetylglucosaminyltransferases (e.g., GnTI, GnTII, and GnTIV) involved in early remodeling of glycans in the mammalian glycosylation pathway. 29162910 1 71 theme glycan 251:256 arg1 microarrays 258:268 glycan microarrays 251:268 glycan microarrays 251:268 Synthesis of homogenous glycans in quantitative yields represents a major bottleneck to the production of molecular tools for glycoscience, such as glycan microarrays, affinity resins, and reference standards. 25361541 9 0 from presentation 1857:1868 arg1 brain 1907:1911 the brain 1903:1911 the brain 1903:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 4 1 theme X 585:585 arg1 epitope 587:593 The Lewis X epitope 575:593 The Lewis X epitope 575:593 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 3 2 theme developing 506:515 arg1 brain 517:521 a developing brain 504:521 a developing brain 504:521 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 9 3 from brain 1907:1911 arg1 presentation 1857:1868 the presentation 1853:1868 the presentation of functional glycan epitopes in the brain 1853:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 9 4 gly O-mannosylated 1627:1640 arg1 glycans 1642:1648 O-mannosylated glycans 1627:1648 O-mannosylated glycans on phosphacan/RPTPβ 1627:1668 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 5 5 theme scaffolding 750:760 arg1 structure 769:777 the scaffolding glycan structure 746:777 the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein 746:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 3 6 theme important 533:541 arg1 roles 543:547 important roles 533:547 important roles 533:547 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 9 7 theme human 1689:1693 arg1 killer-1 1703:1710 human natural killer-1 1689:1710 human natural killer-1 1689:1710 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 9 7 theme human 1689:1693 arg1 epitope 1728:1734 another glycan epitope 1713:1734 another glycan epitope specifically expressed in the nervous system 1713:1779 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 4 8 theme Galβ1-4 628:634 arg1 GlcNAc 646:651 Galβ1-4 (Fucα1-3) GlcNAc 628:651 Galβ1-4 (Fucα1-3) GlcNAc 628:651 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 4 8 theme Galβ1-4 628:634 arg1 trisaccharide 613:625 a trisaccharide 611:625 a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc) 611:652 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 8 9 theme protein 1375:1381 arg1 β1,2-N-acetylglucosaminyltransferase 1393:1428 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 9 theme protein 1375:1381 arg1 N-acetylglucosaminyltransferase 1436:1466 an N-acetylglucosaminyltransferase 1433:1466 an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans 1433:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 7 10 theme Lewis 1276:1280 arg1 epitope 1284:1290 the Lewis X epitope 1272:1290 the Lewis X epitope 1272:1290 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 4 11 theme Fucα1-3 637:643 arg1 GlcNAc 646:651 Galβ1-4 (Fucα1-3) GlcNAc 628:651 Galβ1-4 (Fucα1-3) GlcNAc 628:651 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 4 11 theme Fucα1-3 637:643 arg1 trisaccharide 613:625 a trisaccharide 611:625 a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc) 611:652 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 5 12 theme X 822:822 arg1 epitope 824:830 the Lewis X epitope 812:830 the Lewis X epitope as well as its major carrier protein 812:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 8 13 theme β1,2-N-acetylglucosaminyltransferase 1393:1428 arg1 knockout 1363:1370 knockout 1363:1370 knockout 1363:1370 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 7 14 theme gene-deficient 1176:1189 arg1 mice 1191:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 5 15 theme major 847:851 arg1 protein 861:867 its major carrier protein 843:867 the Lewis X epitope as well as its major carrier protein 812:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 6 16 theme X 955:955 arg1 epitope 957:963 the Lewis X epitope 945:963 the Lewis X epitope 945:963 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 7 17 theme β4GalT2 1167:1173 arg1 mice 1191:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 1 18 theme major 160:164 arg1 modification 174:185 a major protein modification 158:185 a major protein modification 158:185 Glycosylation is a major protein modification. 25361541 1 18 theme major 160:164 arg1 Glycosylation 141:153 Glycosylation 141:153 Glycosylation 141:153 Glycosylation is a major protein modification. 25361541 2 19 located detected 386:393 arg1 number 408:413 a limited number 398:413 a limited number of proteins 398:425 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 2 19 located detected 386:393 arg2 epitope 364:370 a certain glycan epitope 347:370 a certain glycan epitope 347:370 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 0 20 theme developing 75:84 arg1 brain 86:90 the developing brain 71:90 the developing brain 71:90 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 8 21 theme O-mannosylated 1499:1512 arg1 glycans 1514:1520 O-mannosylated glycans 1499:1520 O-mannosylated glycans 1499:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 2 22 theme proteins 418:425 arg1 number 408:413 a limited number 398:413 a limited number of proteins 398:425 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 8 23 gly O-mannosylated 1548:1561 arg1 responsible 1573:1583 responsible 1573:1583 responsible 1573:1583 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 23 gly O-mannosylated 1548:1561 arg1 glycan 1563:1568 the O-mannosylated glycan 1544:1568 the O-mannosylated glycan 1544:1568 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 0 24 from expression 34:43 arg1 brain 86:90 the developing brain 71:90 the developing brain 71:90 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 4 25 theme epitope 699:705 arg1 expression 680:689 the expression 676:689 the expression of this epitope 676:705 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 0 26 theme Major 0:4 arg1 structure 13:21 Major glycan structure 0:21 Major glycan structure underlying expression of the Lewis X epitope in the developing brain 0:90 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 6 27 theme tyrosine 1017:1024 arg1 phosphatase 1026:1036 phosphacan/receptor protein tyrosine phosphatase β 989:1038 phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) 989:1046 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 6 27 theme tyrosine 1017:1024 arg1 RPTPβ 1041:1045 RPTPβ 1041:1045 RPTPβ 1041:1045 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 9 28 theme O-mannosylated 1821:1834 arg1 chains 1843:1848 O-mannosylated glycan chains 1821:1848 O-mannosylated glycan chains 1821:1848 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 8 29 theme X 1323:1323 arg1 epitope 1325:1331 the Lewis X epitope 1313:1331 the Lewis X epitope 1313:1331 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 7 30 theme epitope 1105:1111 arg1 Expression 1079:1088 Expression 1079:1088 Expression of the Lewis X epitope 1079:1111 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 5 31 theme nervous 909:915 arg1 system 917:922 the nervous system 905:922 the nervous system 905:922 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 6 32 theme phosphacan/receptor 989:1007 arg1 phosphatase 1026:1036 phosphacan/receptor protein tyrosine phosphatase β 989:1038 phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) 989:1046 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 6 32 theme phosphacan/receptor 989:1007 arg1 RPTPβ 1041:1045 RPTPβ 1041:1045 RPTPβ 1041:1045 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 9 33 theme chains 1843:1848 arg1 importance 1807:1816 the importance 1803:1816 the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain 1803:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 7 34 theme Lewis 1097:1101 arg1 epitope 1105:1111 the Lewis X epitope 1093:1111 the Lewis X epitope 1093:1111 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 3 35 theme glycan 437:442 arg1 epitopes 444:451 these glycan epitopes 431:451 these glycan epitopes 431:451 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 0 36 theme X 58:58 arg1 epitope 60:66 the Lewis X epitope 48:66 the Lewis X epitope 48:66 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 0 37 link O-mannose-linked 95:110 arg1 glycans 112:118 O-mannose-linked glycans 95:118 O-mannose-linked glycans on phosphacan/RPTPβ 95:138 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 6 38 theme developing 1055:1064 arg1 brain 1072:1076 the developing mouse brain 1051:1076 the developing mouse brain 1051:1076 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 9 39 theme glycan 1884:1889 arg1 epitopes 1891:1898 functional glycan epitopes 1873:1898 functional glycan epitopes in the brain 1873:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 3 40 theme brain 517:521 arg1 stage 495:499 the early stage 485:499 the early stage of a developing brain 485:521 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 5 41 theme glycan 762:767 arg1 structure 769:777 the scaffolding glycan structure 746:777 the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein 746:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 9 42 theme O-mannosylated 1627:1640 arg1 glycans 1642:1648 O-mannosylated glycans 1627:1648 O-mannosylated glycans on phosphacan/RPTPβ 1627:1668 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 8 43 theme Lewis 1604:1608 arg1 epitope 1612:1618 the Lewis X epitope 1600:1618 the Lewis X epitope 1600:1618 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 4 44 theme Lewis 579:583 arg1 epitope 587:593 The Lewis X epitope 575:593 The Lewis X epitope 575:593 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 2 45 theme Golgi 330:334 arg1 apparatus 336:344 the Golgi apparatus 326:344 the Golgi apparatus 326:344 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 1 46 theme protein 166:172 arg1 modification 174:185 a major protein modification 158:185 a major protein modification 158:185 Glycosylation is a major protein modification. 25361541 1 46 theme protein 166:172 arg1 Glycosylation 141:153 Glycosylation 141:153 Glycosylation 141:153 Glycosylation is a major protein modification. 25361541 9 47 from importance 1807:1816 arg1 presentation 1857:1868 the presentation 1853:1868 the presentation of functional glycan epitopes in the brain 1853:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 2 48 theme certain 349:355 arg1 epitope 364:370 a certain glycan epitope 347:370 a certain glycan epitope 347:370 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 9 49 theme natural 1695:1701 arg1 killer-1 1703:1710 human natural killer-1 1689:1710 human natural killer-1 1689:1710 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 9 49 theme natural 1695:1701 arg1 epitope 1728:1734 another glycan epitope 1713:1734 another glycan epitope specifically expressed in the nervous system 1713:1779 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 9 50 from epitopes 1891:1898 arg1 brain 1907:1911 the brain 1903:1911 the brain 1903:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 3 51 theme cell-cell 552:560 arg1 interaction 562:572 cell-cell interaction 552:572 cell-cell interaction 552:572 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 8 52 theme O-mannose 1383:1391 arg1 β1,2-N-acetylglucosaminyltransferase 1393:1428 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 52 theme O-mannose 1383:1391 arg1 N-acetylglucosaminyltransferase 1436:1466 an N-acetylglucosaminyltransferase 1433:1466 an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans 1433:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 5 53 theme protein 861:867 arg1 formation 799:807 the formation 795:807 the formation of the Lewis X epitope as well as its major carrier protein 795:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 5 54 theme Lewis 816:820 arg1 epitope 824:830 the Lewis X epitope 812:830 the Lewis X epitope as well as its major carrier protein 812:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 9 55 theme glycan 1721:1726 arg1 killer-1 1703:1710 human natural killer-1 1689:1710 human natural killer-1 1689:1710 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 9 55 theme glycan 1721:1726 arg1 epitope 1728:1734 another glycan epitope 1713:1734 another glycan epitope specifically expressed in the nervous system 1713:1779 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 2 56 theme several 244:250 arg1 glycosyltransferases 252:271 several glycosyltransferases 244:271 several glycosyltransferases 244:271 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 9 57 from glycans 1642:1648 arg1 phosphacan/RPTPβ 1653:1668 phosphacan/RPTPβ 1653:1668 phosphacan/RPTPβ 1653:1668 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 2 58 theme endoplasmic 301:311 arg1 reticulum 313:321 the endoplasmic reticulum 297:321 the endoplasmic reticulum 297:321 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 2 59 from reticulum 313:321 arg1 trafficking 280:290 trafficking 280:290 trafficking from the endoplasmic reticulum to the Golgi apparatus 280:344 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 8 60 theme essential 1468:1476 arg1 β1,2-N-acetylglucosaminyltransferase 1393:1428 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 1375:1430 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 60 theme essential 1468:1476 arg1 N-acetylglucosaminyltransferase 1436:1466 an N-acetylglucosaminyltransferase 1433:1466 an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans 1433:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 0 61 theme epitope 60:66 arg1 expression 34:43 expression 34:43 expression of the Lewis X epitope in the developing brain 34:90 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 8 62 gly O-mannosylated 1499:1512 arg1 glycans 1514:1520 O-mannosylated glycans 1499:1520 O-mannosylated glycans 1499:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 9 63 theme nervous 1766:1772 arg1 system 1774:1779 the nervous system 1762:1779 the nervous system 1762:1779 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 5 64 theme carrier 853:859 arg1 protein 861:867 its major carrier protein 843:867 the Lewis X epitope as well as its major carrier protein 812:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 6 65 theme Lewis 949:953 arg1 epitope 957:963 the Lewis X epitope 945:963 the Lewis X epitope 945:963 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 4 66 theme key 661:663 arg1 enzyme 665:670 a key enzyme 659:670 a key enzyme for the expression of this epitope 659:705 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 4 66 theme key 661:663 arg1 α1,3-fucosyltransferase 710:732 α1,3-fucosyltransferase 9 710:734 α1,3-fucosyltransferase 9 710:734 The Lewis X epitope is comprised of a trisaccharide (Galβ1-4 (Fucα1-3) GlcNAc), and a key enzyme for the expression of this epitope is α1,3-fucosyltransferase 9. 25361541 7 67 theme β1,4-galactosyltransferase 1137:1162 arg1 mice 1191:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice 1137:1194 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 8 68 theme glycans 1514:1520 arg1 synthesis 1486:1494 the synthesis 1482:1494 the synthesis of O-mannosylated glycans 1482:1520 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 2 69 theme glycan 357:362 arg1 epitope 364:370 a certain glycan epitope 347:370 a certain glycan epitope 347:370 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 5 70 theme epitope 824:830 arg1 formation 799:807 the formation 795:807 the formation of the Lewis X epitope as well as its major carrier protein 795:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 25361541 0 71 theme O-mannose-linked 95:110 arg1 glycans 112:118 O-mannose-linked glycans 95:118 O-mannose-linked glycans on phosphacan/RPTPβ 95:138 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 9 72 gly O-mannosylated 1821:1834 arg1 chains 1843:1848 O-mannosylated glycan chains 1821:1848 O-mannosylated glycan chains 1821:1848 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 7 73 theme X 1282:1282 arg1 epitope 1284:1290 the Lewis X epitope 1272:1290 the Lewis X epitope 1272:1290 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 0 74 theme glycan 6:11 arg1 structure 13:21 Major glycan structure 0:21 Major glycan structure underlying expression of the Lewis X epitope in the developing brain 0:90 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 2 75 theme limited 400:406 arg1 number 408:413 a limited number 398:413 a limited number of proteins 398:425 Although proteins are glycosylated/further modulated by several glycosyltransferases during trafficking from the endoplasmic reticulum to the Golgi apparatus, a certain glycan epitope has only been detected on a limited number of proteins. 25361541 7 76 theme X 1103:1103 arg1 epitope 1105:1111 the Lewis X epitope 1093:1111 the Lewis X epitope 1093:1111 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 6 77 theme protein 1009:1015 arg1 phosphatase 1026:1036 phosphacan/receptor protein tyrosine phosphatase β 989:1038 phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) 989:1046 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 6 77 theme protein 1009:1015 arg1 RPTPβ 1041:1045 RPTPβ 1041:1045 RPTPβ 1041:1045 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 9 78 theme glycan 1836:1841 arg1 chains 1843:1848 O-mannosylated glycan chains 1821:1848 O-mannosylated glycan chains 1821:1848 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 8 79 theme O-mannosylated 1548:1561 arg1 responsible 1573:1583 responsible 1573:1583 responsible 1573:1583 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 79 theme O-mannosylated 1548:1561 arg1 glycan 1563:1568 the O-mannosylated glycan 1544:1568 the O-mannosylated glycan 1544:1568 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 8 80 theme Lewis 1317:1321 arg1 epitope 1325:1331 the Lewis X epitope 1313:1331 the Lewis X epitope 1313:1331 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 7 81 theme major 1231:1235 arg1 β4GalT2 1218:1224 β4GalT2 1218:1224 β4GalT2 1218:1224 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 7 81 theme major 1231:1235 arg1 galactosyltransferase 1237:1257 a major galactosyltransferase 1229:1257 a major galactosyltransferase required for the Lewis X epitope 1229:1290 Expression of the Lewis X epitope was markedly reduced in β1,4-galactosyltransferase 2 (β4GalT2) gene-deficient mice, which indicated that β4GalT2 is a major galactosyltransferase required for the Lewis X epitope. 25361541 3 82 theme Lewis 454:458 arg1 X 460:460 Lewis X 454:460 Lewis X 454:460 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 0 83 theme Lewis 52:56 arg1 epitope 60:66 the Lewis X epitope 48:66 the Lewis X epitope 48:66 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 6 84 theme mouse 1066:1070 arg1 brain 1072:1076 the developing mouse brain 1051:1076 the developing mouse brain 1051:1076 Here we showed that the Lewis X epitope was mainly expressed on phosphacan/receptor protein tyrosine phosphatase β (RPTPβ) in the developing mouse brain. 25361541 9 85 theme functional 1873:1882 arg1 epitopes 1891:1898 functional glycan epitopes 1873:1898 functional glycan epitopes in the brain 1873:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 3 86 theme early 489:493 arg1 stage 495:499 the early stage 485:499 the early stage of a developing brain 485:521 Of these glycan epitopes, Lewis X is highly expressed in the early stage of a developing brain and plays important roles in cell-cell interaction. 25361541 9 87 theme epitopes 1891:1898 arg1 presentation 1857:1868 the presentation 1853:1868 the presentation of functional glycan epitopes in the brain 1853:1911 Since O-mannosylated glycans on phosphacan/RPTPβ could also present human natural killer-1, another glycan epitope specifically expressed in the nervous system, our results revealed the importance of O-mannosylated glycan chains in the presentation of functional glycan epitopes in the brain. 25361541 8 88 theme X 1610:1610 arg1 epitope 1612:1618 the Lewis X epitope 1600:1618 the Lewis X epitope 1600:1618 We also showed that the Lewis X epitope almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for the synthesis of O-mannosylated glycans, which indicated that the O-mannosylated glycan is responsible for presenting the Lewis X epitope. 25361541 0 89 from glycans 112:118 arg1 phosphacan/RPTPβ 123:138 phosphacan/RPTPβ 123:138 phosphacan/RPTPβ 123:138 Major glycan structure underlying expression of the Lewis X epitope in the developing brain is O-mannose-linked glycans on phosphacan/RPTPβ. 25361541 5 90 theme responsible 779:789 arg1 structure 769:777 the scaffolding glycan structure 746:777 the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein 746:867 However, the scaffolding glycan structure responsible for the formation of the Lewis X epitope as well as its major carrier protein has not been fully characterized in the nervous system. 28025252 5 0 theme mass 728:731 arg1 spectrometry 733:744 mass spectrometry 728:744 mass spectrometry 728:744 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 4 1 from Comparison 470:479 arg1 controls 513:520 20 healthy controls 502:520 20 healthy controls 502:520 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 7 2 theme 6-sulfo 924:930 arg1 lactosamine 938:948 6-sulfo type2 lactosamine 924:948 6-sulfo type2 lactosamine 924:948 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 7 2 theme 6-sulfo 924:930 arg1 A. 1031:1032 A. 1031:1032 A. 1031:1032 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 3 theme type1-type2 1419:1429 arg1 backbones 1438:1446 type1-type2 hybrid backbones 1419:1446 type1-type2 hybrid backbones with 6-sulfo Lewis X 1419:1467 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 1 4 theme sulfated 124:131 arg1 O-glycans 133:141 sulfated O-glycans 124:141 sulfated O-glycans 124:141 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 1 5 theme tumor 194:198 arg1 markers 200:206 novel serum carbohydrate tumor markers 169:206 novel serum carbohydrate tumor markers 169:206 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 7 6 theme various 895:901 arg1 lactosamine 938:948 6-sulfo type2 lactosamine 924:948 6-sulfo type2 lactosamine 924:948 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 7 6 theme various 895:901 arg1 determinants 903:914 various determinants 895:914 various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone 895:1142 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 7 theme 6-sulfo 1453:1459 arg1 X 1467:1467 6-sulfo Lewis X 1453:1467 6-sulfo Lewis X 1453:1467 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 8 theme candidates 1659:1668 arg1 analysis 1638:1645 analysis 1638:1645 analysis of multiple candidates 1638:1668 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 8 theme candidates 1659:1668 arg1 means 1691:1695 an effective means 1678:1695 an effective means of screening various cancers 1678:1724 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 4 9 from controls 513:520 arg1 profiles 488:495 the profiles 484:495 the profiles from 20 healthy controls 484:520 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 4 9 from controls 513:520 arg1 Comparison 470:479 Comparison 470:479 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients 470:578 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 2 10 theme anion 328:332 arg1 chromatography 343:356 anion exchange chromatography 328:356 anion exchange chromatography 328:356 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 8 11 theme reaction 1249:1256 arg1 assay 1269:1273 a selected reaction monitoring assay 1238:1273 a selected reaction monitoring assay 1238:1273 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 9 12 theme 6-sulfo 1470:1476 arg1 lactosamine 1484:1494 6-sulfo type2 lactosamine or 3'-sulfo Lewis A 1470:1514 lactosamine 1484:1494 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 13 theme effective 1681:1689 arg1 means 1691:1695 an effective means 1678:1695 an effective means of screening various cancers 1678:1724 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 13 theme effective 1681:1689 arg1 analysis 1638:1645 analysis 1638:1645 analysis of multiple candidates 1638:1668 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 10 14 theme further 1789:1795 arg1 study 1810:1814 a further verification study 1787:1814 a further verification study 1787:1814 To fully evaluate the clinical utility of these candidates, a further verification study is required. 28025252 9 15 theme 3'-sulfo 1499:1506 arg1 A 1514:1514 6-sulfo type2 lactosamine or 3'-sulfo Lewis A 1470:1514 A 1514:1514 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 16 theme various 1710:1716 arg1 cancers 1718:1724 screening various cancers 1700:1724 screening various cancers 1700:1724 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 4 17 theme profiles 488:495 arg1 Comparison 470:479 Comparison 470:479 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients 470:578 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 0 18 theme focused 68:74 arg1 analyses 85:92 focused glycomic analyses 68:92 focused glycomic analyses 68:92 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 2 19 theme pyridylaminated 272:286 arg1 glycans 288:294 pyridylaminated glycans 272:294 pyridylaminated glycans prepared from sera 272:313 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 3 20 theme Sulfated 359:366 arg1 profiles 377:384 Sulfated O-glycan profiles 359:384 Sulfated O-glycan profiles 359:384 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 1 21 theme serum 175:179 arg1 markers 200:206 novel serum carbohydrate tumor markers 169:206 novel serum carbohydrate tumor markers 169:206 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 4 22 theme pancreatic 553:562 arg1 patients 571:578 11 gastric and 9 pancreatic cancer patients 536:578 11 gastric and 9 pancreatic cancer patients 536:578 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 9 23 theme candidates 1323:1332 arg1 levels 1309:1314 the levels 1305:1314 the levels of all candidates 1305:1332 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 8 24 from levels 1149:1154 arg1 sera 1190:1193 the sera 1186:1193 the sera from all 40 subjects 1186:1214 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 8 24 from levels 1149:1154 arg1 subjects 1207:1214 all 40 subjects 1200:1214 all 40 subjects 1200:1214 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 7 25 theme 3'-sulfo 985:992 arg1 lactosamine 1000:1010 3'-sulfo type1 lactosamine 985:1010 3'-sulfo type1 lactosamine 985:1010 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 26 theme variety 1541:1547 arg1 sera 1533:1536 sera 1533:1536 sera of variety of patients 1533:1559 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 4 27 with Comparison 470:479 arg1 those 527:531 those 527:531 those 527:531 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 0 28 theme Various 0:6 arg1 candidates 43:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 9 29 dep candidates 1401:1410 arg1 ii 1391:1392 ii 1391:1392 ii 1391:1392 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 5 30 theme candidates 637:646 arg1 structures 617:626 The structures 613:626 The structures of these candidates 613:646 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 9 31 theme patients 1552:1559 arg1 patients 1552:1559 patients 1552:1559 patients 1552:1559 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 31 theme patients 1552:1559 arg1 variety 1541:1547 variety 1541:1547 variety of patients 1541:1559 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 0 32 theme carbohydrate 17:28 arg1 candidates 43:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 9 33 theme hybrid 1431:1436 arg1 backbones 1438:1446 type1-type2 hybrid backbones 1419:1446 type1-type2 hybrid backbones with 6-sulfo Lewis X 1419:1467 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 0 34 theme marker 36:41 arg1 candidates 43:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 3 35 theme chromatography 443:456 arg1 separation 458:467 high performance liquid chromatography separation 419:467 high performance liquid chromatography separation 419:467 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 8 36 from subjects 1207:1214 arg1 sera 1190:1193 the sera 1186:1193 the sera from all 40 subjects 1186:1214 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 8 36 from subjects 1207:1214 arg1 levels 1149:1154 The levels 1145:1154 The levels of these candidate glycans in the sera from all 40 subjects 1145:1214 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 7 37 theme Lewis 976:980 arg1 Y 982:982 6-sulfo Lewis Y 968:982 6-sulfo Lewis Y 968:982 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 2 38 theme Sulfated 209:216 arg1 glycans 218:224 Sulfated glycans 209:224 Sulfated glycans 209:224 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 8 39 theme glycans 1175:1181 arg1 levels 1149:1154 The levels 1145:1154 The levels of these candidate glycans in the sera from all 40 subjects 1145:1214 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 7 40 theme Lewis 959:963 arg1 X 965:965 6-sulfo Lewis X 951:965 6-sulfo Lewis X 951:965 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 41 theme core1 1395:1399 arg1 candidates 1401:1410 (ii) core1 candidates 1390:1410 (ii) core1 candidates 1390:1410 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 7 42 theme hybrid 1128:1133 arg1 backbone 1135:1142 type2 lactosamine hybrid backbone 1110:1142 type2 lactosamine hybrid backbone 1110:1142 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 7 43 theme type2 932:936 arg1 lactosamine 938:948 6-sulfo type2 lactosamine 924:948 6-sulfo type2 lactosamine 924:948 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 7 43 theme type2 932:936 arg1 A. 1031:1032 A. 1031:1032 A. 1031:1032 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 1 44 theme carbohydrate 181:192 arg1 markers 200:206 novel serum carbohydrate tumor markers 169:206 novel serum carbohydrate tumor markers 169:206 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 7 45 theme lactosamine 1116:1126 arg1 backbone 1135:1142 type2 lactosamine hybrid backbone 1110:1142 type2 lactosamine hybrid backbone 1110:1142 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 46 dep levels 1572:1577 arg1 elevated 1339:1346 elevated 1339:1346 were elevated in sera of at least one or more patients 1334:1387 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 46 dep levels 1572:1577 arg1 iii 1567:1569 iii 1567:1569 iii 1567:1569 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 46 dep levels 1572:1577 arg1 elevated 1521:1528 elevated 1521:1528 were elevated in sera of variety of patients 1516:1559 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 5 47 theme enzymatic 704:712 arg1 digestion 714:722 enzymatic digestion 704:722 enzymatic digestion 704:722 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 10 48 theme clinical 1749:1756 arg1 utility 1758:1764 the clinical utility 1745:1764 the clinical utility of these candidates 1745:1784 To fully evaluate the clinical utility of these candidates, a further verification study is required. 28025252 9 49 theme multiple 1650:1657 arg1 candidates 1659:1668 multiple candidates 1650:1668 multiple candidates 1650:1668 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 7 50 theme type2 1110:1114 arg1 backbone 1135:1142 type2 lactosamine hybrid backbone 1110:1142 type2 lactosamine hybrid backbone 1110:1142 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 2 51 theme exchange 334:341 arg1 chromatography 343:356 anion exchange chromatography 328:356 anion exchange chromatography 328:356 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 9 52 theme Lewis 1461:1465 arg1 X 1467:1467 6-sulfo Lewis X 1453:1467 6-sulfo Lewis X 1453:1467 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 4 53 theme healthy 505:511 arg1 controls 513:520 20 healthy controls 502:520 20 healthy controls 502:520 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 10 54 theme candidates 1775:1784 arg1 utility 1758:1764 the clinical utility 1745:1764 the clinical utility of these candidates 1745:1784 To fully evaluate the clinical utility of these candidates, a further verification study is required. 28025252 8 55 theme selected 1240:1247 arg1 assay 1269:1273 a selected reaction monitoring assay 1238:1273 a selected reaction monitoring assay 1238:1273 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 2 56 theme α-neuraminidase 243:257 arg1 digestion 259:267 α-neuraminidase digestion 243:267 α-neuraminidase digestion of pyridylaminated glycans prepared from sera 243:313 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 9 57 theme type2 1478:1482 arg1 lactosamine 1484:1494 6-sulfo type2 lactosamine or 3'-sulfo Lewis A 1470:1514 lactosamine 1484:1494 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 8 58 theme monitoring 1258:1267 arg1 assay 1269:1273 a selected reaction monitoring assay 1238:1273 a selected reaction monitoring assay 1238:1273 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 10 59 theme verification 1797:1808 arg1 study 1810:1814 a further verification study 1787:1814 a further verification study 1787:1814 To fully evaluate the clinical utility of these candidates, a further verification study is required. 28025252 7 60 theme core1 1057:1061 arg1 glycans 1063:1069 the core1 glycans 1053:1069 the core1 glycans 1053:1069 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 61 theme screening 1700:1708 arg1 cancers 1718:1724 screening various cancers 1700:1724 screening various cancers 1700:1724 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 0 62 theme glycomic 76:83 arg1 analyses 85:92 focused glycomic analyses 68:92 focused glycomic analyses 68:92 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 2 63 theme glycans 288:294 arg1 digestion 259:267 α-neuraminidase digestion 243:267 α-neuraminidase digestion of pyridylaminated glycans prepared from sera 243:313 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 9 64 theme Lewis 1508:1512 arg1 A 1514:1514 6-sulfo type2 lactosamine or 3'-sulfo Lewis A 1470:1514 A 1514:1514 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 65 theme cancers 1718:1724 arg1 means 1691:1695 an effective means 1678:1695 an effective means of screening various cancers 1678:1724 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 65 theme cancers 1718:1724 arg1 analysis 1638:1645 analysis 1638:1645 analysis of multiple candidates 1638:1668 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 1 66 theme novel 169:173 arg1 markers 200:206 novel serum carbohydrate tumor markers 169:206 novel serum carbohydrate tumor markers 169:206 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 3 67 theme O-glycan 368:375 arg1 profiles 377:384 Sulfated O-glycan profiles 359:384 Sulfated O-glycan profiles 359:384 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 4 68 theme cancer 564:569 arg1 patients 571:578 11 gastric and 9 pancreatic cancer patients 536:578 11 gastric and 9 pancreatic cancer patients 536:578 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 6 69 theme core2 786:790 arg1 glycans 792:798 9 core1 and 5 core2 glycans 772:798 glycans 792:798 The candidates comprised 9 core1 and 5 core2 glycans. 28025252 5 70 theme various 678:684 arg1 methods 686:692 various methods 678:692 various methods including enzymatic digestion and mass spectrometry 678:744 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 5 70 theme various 678:684 arg1 digestion 714:722 enzymatic digestion 704:722 enzymatic digestion 704:722 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 5 70 theme various 678:684 arg1 spectrometry 733:744 mass spectrometry 728:744 mass spectrometry 728:744 The structures of these candidates were precisely analyzed using various methods including enzymatic digestion and mass spectrometry. 28025252 4 71 theme gastric 539:545 arg1 patients 571:578 11 gastric and 9 pancreatic cancer patients 536:578 11 gastric and 9 pancreatic cancer patients 536:578 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 0 72 theme sulfated 8:15 arg1 candidates 43:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 7 73 theme type1 994:998 arg1 lactosamine 1000:1010 3'-sulfo type1 lactosamine 985:1010 3'-sulfo type1 lactosamine 985:1010 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 1 74 theme Glycomic 95:102 arg1 analysis 104:111 Glycomic analysis 95:111 Glycomic analysis focused on sulfated O-glycans 95:141 Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. 28025252 3 75 theme liquid 436:441 arg1 chromatography 443:456 high performance liquid chromatography 419:456 high performance liquid chromatography separation 419:467 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 7 76 dep A. 1031:1032 arg1 displayed 1088:1096 displayed 1088:1096 displayed a type1 and type2 lactosamine hybrid backbone 1088:1142 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 0 77 theme tumor 30:34 arg1 candidates 43:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates 0:52 Various sulfated carbohydrate tumor marker candidates identified by focused glycomic analyses. 28025252 3 78 theme high 419:422 arg1 chromatography 443:456 high performance liquid chromatography 419:456 high performance liquid chromatography separation 419:467 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 9 79 dep elevated 1339:1346 arg1 i 1302:1302 i 1302:1302 i 1302:1302 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 3 80 theme separation 458:467 arg1 types 410:414 two types 406:414 two types of high performance liquid chromatography separation 406:467 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 4 81 theme marker 594:599 arg1 candidates 601:610 14 marker candidates 591:610 14 marker candidates 591:610 Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates. 28025252 7 82 theme 6-sulfo 968:974 arg1 Y 982:982 6-sulfo Lewis Y 968:982 6-sulfo Lewis Y 968:982 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28025252 9 83 theme patients 1380:1387 arg1 sera 1351:1354 sera 1351:1354 sera of at least one or more patients 1351:1387 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 8 84 theme candidate 1165:1173 arg1 glycans 1175:1181 these candidate glycans 1159:1181 these candidate glycans 1159:1181 The levels of these candidate glycans in the sera from all 40 subjects were quantified using a selected reaction monitoring assay. 28025252 9 85 with backbones 1438:1446 arg1 X 1467:1467 6-sulfo Lewis X 1453:1467 6-sulfo Lewis X 1453:1467 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 9 86 theme candidates 1586:1595 arg1 levels 1572:1577 : (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels 1299:1577 : (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients 1299:1624 These analyses revealed: (i) the levels of all candidates were elevated in sera of at least one or more patients; (ii) core1 candidates having type1-type2 hybrid backbones with 6-sulfo Lewis X, 6-sulfo type2 lactosamine or 3'-sulfo Lewis A were elevated in sera of variety of patients; and (iii) levels of the candidates varied widely among patients, suggesting analysis of multiple candidates will be an effective means of screening various cancers. 28025252 2 87 dep enriched 231:238 arg1 followed 316:323 followed 316:323 followed by anion exchange chromatography 316:356 Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. 28025252 3 88 theme performance 424:434 arg1 chromatography 443:456 high performance liquid chromatography 419:456 high performance liquid chromatography separation 419:467 Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. 28025252 7 89 theme 6-sulfo 951:957 arg1 X 965:965 6-sulfo Lewis X 951:965 6-sulfo Lewis X 951:965 All these candidates were monosulfated, and 11 were also mono- or difucosylated, and included various determinants such as 6-sulfo type2 lactosamine, 6-sulfo Lewis X, 6-sulfo Lewis Y, 3'-sulfo type1 lactosamine and 3'-sulfo Lewis A. Furthermore, among the core1 glycans, five candidates displayed a type1 and type2 lactosamine hybrid backbone. 28054185 5 0 theme sialic 720:725 arg1 NANA 761:764 NANA 761:764 NANA 761:764 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 5 0 theme sialic 720:725 arg1 acid 727:730 sialic acid 720:730 sialic acid 720:730 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 4 1 theme CHO 667:669 arg1 cells 671:675 CHO cells 667:675 CHO cells (37%) 667:681 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 4 1 theme CHO 667:669 arg1 % 680:680 37% 678:680 37% 678:680 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 5 2 theme acid 727:730 arg1 residues 767:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 6 3 theme linked 855:860 arg1 forms 874:878 only α-2,3 linked sialic acid forms 844:878 only α-2,3 linked sialic acid forms 844:878 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 8 4 theme specific 1125:1132 arg1 N-glycan 1134:1141 human specific N-glycan 1119:1141 human specific N-glycan 1119:1141 CONCLUSIONS F2N78 cells stably produced antibodies with human specific N-glycan. 28054185 2 5 theme culture 329:335 arg1 performance 337:347 The culture performance 325:347 The culture performance 325:347 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 1 6 theme physicochemical 227:241 arg1 properties 243:252 physicochemical properties 227:252 physicochemical properties 227:252 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 0 7 theme culture 75:81 arg1 performances 83:94 culture performances 75:94 culture performances 75:94 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 5 8 link α-2,6-linked 707:718 arg1 residues 767:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 5 9 theme sialic 688:693 arg1 forms 700:704 sialic acid forms 688:704 sialic acid forms 688:704 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 9 10 theme novel 1148:1152 arg1 system 1165:1170 The novel expression system 1144:1170 The novel expression system based on human cells 1144:1191 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 5 11 theme acid 695:698 arg1 forms 700:704 sialic acid forms 688:704 sialic acid forms 688:704 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 9 12 theme recombinant 1270:1280 arg1 proteins 1282:1289 recombinant proteins 1270:1289 recombinant proteins 1270:1289 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 1 13 with comparison 290:299 arg1 cells 310:314 CHO cells 306:314 CHO cells 306:314 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 3 14 theme acidic 521:526 arg1 variants 535:542 a greater acidic charge variants 511:542 a greater acidic charge variants 511:542 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 6 15 link linked 855:860 arg1 forms 874:878 only α-2,3 linked sialic acid forms 844:878 only α-2,3 linked sialic acid forms 844:878 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 4 16 theme F2N78 643:647 arg1 cells 649:653 F2N78 cells 643:653 F2N78 cells 643:653 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 0 17 theme performances 83:94 arg1 comparison 61:70 a comparison 59:70 a comparison of culture performances and protein qualities 59:116 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 9 18 theme proteins 1282:1289 arg1 production 1256:1265 production 1256:1265 production of recombinant proteins 1256:1289 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 7 19 theme F2N78 1050:1054 arg1 cells 1056:1060 F2N78 cells 1050:1060 F2N78 cells 1050:1060 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 3 20 theme charge 528:533 arg1 variants 535:542 a greater acidic charge variants 511:542 a greater acidic charge variants 511:542 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 2 21 theme antibodies 383:392 arg1 performance 337:347 The culture performance 325:347 The culture performance 325:347 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 2 21 theme antibodies 383:392 arg1 properties 369:378 physicochemical properties 353:378 physicochemical properties of antibodies produced from F2N78 and CHO cells 353:426 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 1 22 theme novel 167:171 arg1 line 184:187 a novel human cell line 165:187 a novel human cell line 165:187 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 22 theme novel 167:171 arg1 F2N78 190:194 F2N78 190:194 F2N78 190:194 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 6 23 theme CHO 922:924 arg1 cells 926:930 CHO cells 922:930 CHO cells 922:930 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 9 24 theme host 1242:1245 arg1 cell 1247:1250 an alternative host cell 1227:1250 an alternative host cell 1227:1250 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 1 25 theme human 173:177 arg1 line 184:187 a novel human cell line 165:187 a novel human cell line 165:187 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 25 theme human 173:177 arg1 F2N78 190:194 F2N78 190:194 F2N78 190:194 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 0 26 theme hybrid 26:31 arg1 line 38:41 human hybrid cell line 20:41 human hybrid cell line 20:41 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 0 26 theme hybrid 26:31 arg1 F2N78 44:48 F2N78 44:48 F2N78 44:48 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 1 27 theme biological 259:268 arg1 activity 270:277 biological activity 259:277 biological activity 259:277 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 0 28 theme qualities 108:116 arg1 comparison 61:70 a comparison 59:70 a comparison of culture performances and protein qualities 59:116 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 9 29 theme expression 1154:1163 arg1 system 1165:1170 The novel expression system 1144:1170 The novel expression system based on human cells 1144:1191 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 3 30 theme CHO 549:551 arg1 cells 553:557 CHO cells 549:557 CHO cells 549:557 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 8 31 with antibodies 1103:1112 arg1 N-glycan 1134:1141 human specific N-glycan 1119:1141 human specific N-glycan 1119:1141 CONCLUSIONS F2N78 cells stably produced antibodies with human specific N-glycan. 28054185 2 32 theme CHO 418:420 arg1 cells 422:426 CHO cells 418:426 CHO cells 418:426 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 0 33 theme human 20:24 arg1 line 38:41 human hybrid cell line 20:41 human hybrid cell line 20:41 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 0 33 theme human 20:24 arg1 F2N78 44:48 F2N78 44:48 F2N78 44:48 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 0 34 theme protein 100:106 arg1 qualities 108:116 protein qualities 100:116 protein qualities 100:116 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 2 35 dep RESULTS 317:323 arg1 performance 337:347 The culture performance 325:347 The culture performance 325:347 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 2 35 dep RESULTS 317:323 arg1 properties 369:378 physicochemical properties 353:378 physicochemical properties of antibodies produced from F2N78 and CHO cells 353:426 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 6 36 theme sialic 862:867 arg1 forms 874:878 only α-2,3 linked sialic acid forms 844:878 only α-2,3 linked sialic acid forms 844:878 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 1 37 theme line 184:187 arg1 activity 270:277 biological activity 259:277 biological activity 259:277 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 37 theme line 184:187 arg1 performance 214:224 growth performance 207:224 growth performance 207:224 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 37 theme line 184:187 arg1 properties 243:252 physicochemical properties 227:252 physicochemical properties 227:252 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 37 theme line 184:187 arg1 characteristics 146:160 the characteristics 142:160 the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity 142:277 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 0 38 theme line 38:41 arg1 Characterization 0:15 Characterization 0:15 Characterization of human hybrid cell line, F2N78, 0:49 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 6 39 located detected 885:892 arg2 forms 874:878 only α-2,3 linked sialic acid forms 844:878 only α-2,3 linked sialic acid forms 844:878 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 6 39 located detected 885:892 arg1 contrast 834:841 contrast 834:841 contrast 834:841 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 6 39 located detected 885:892 arg1 antibodies 897:906 antibodies 897:906 antibodies produced from CHO cells 897:930 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 4 40 from % 619:619 arg1 CT-A 624:627 CT-A 624:627 CT-A 624:627 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 0 41 theme cell 33:36 arg1 line 38:41 human hybrid cell line 20:41 human hybrid cell line 20:41 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 0 41 theme cell 33:36 arg1 F2N78 44:48 F2N78 44:48 F2N78 44:48 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 1 42 theme direct 283:288 arg1 comparison 290:299 direct comparison 283:299 direct comparison with CHO cells 283:314 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 4 43 theme main 570:573 arg1 glycoforms 575:584 main glycoforms 570:584 main glycoforms 570:584 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 6 44 theme N-glycolylneuraminic 946:965 arg1 acid 967:970 N-glycolylneuraminic acid 946:970 N-glycolylneuraminic acid 946:970 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 3 45 theme charge 447:452 arg1 variants 454:461 charge variants 447:461 charge variants 447:461 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 5 46 theme acid 755:758 arg1 residues 767:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 8 47 theme human 1119:1123 arg1 N-glycan 1134:1141 human specific N-glycan 1119:1141 human specific N-glycan 1119:1141 CONCLUSIONS F2N78 cells stably produced antibodies with human specific N-glycan. 28054185 7 48 theme Hybrid 987:992 arg1 structure 994:1002 Hybrid structure 987:1002 Hybrid structure 987:1002 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 9 49 theme human 1181:1185 arg1 cells 1187:1191 human cells 1181:1191 human cells 1181:1191 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 7 50 theme bisecting 1008:1016 arg1 structure 1018:1026 bisecting structure 1008:1026 bisecting structure 1008:1026 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 6 51 theme acid 869:872 arg1 forms 874:878 only α-2,3 linked sialic acid forms 844:878 only α-2,3 linked sialic acid forms 844:878 In contrast, only α-2,3 linked sialic acid forms were detected in antibodies produced from CHO cells, and NANA and N-glycolylneuraminic acid were detected. 28054185 4 52 dep % 619:619 arg1 compared 655:662 compared 655:662 compared to CHO cells (37%) 655:681 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 9 53 theme cell 1247:1250 arg1 development 1212:1222 the development 1208:1222 the development of an alternative host cell for production of recombinant proteins 1208:1289 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 3 54 theme F2N78 489:493 arg1 cells 495:499 F2N78 cells 489:499 F2N78 cells 489:499 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 1 55 theme CHO 306:308 arg1 cells 310:314 CHO cells 306:314 CHO cells 306:314 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 8 56 theme F2N78 1075:1079 arg1 cells 1081:1085 CONCLUSIONS F2N78 cells 1063:1085 CONCLUSIONS F2N78 cells 1063:1085 CONCLUSIONS F2N78 cells stably produced antibodies with human specific N-glycan. 28054185 5 57 located observed 781:788 arg1 antibodies 793:802 antibodies 793:802 antibodies produced from F2N78 cells 793:828 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 5 57 located observed 781:788 arg2 residues 767:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 5 58 theme N-acetylneuraminic 736:753 arg1 acid 755:758 N-acetylneuraminic acid 736:758 N-acetylneuraminic acid 736:758 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 4 59 theme galactosylation 597:611 arg1 degree 587:592 degree 587:592 degree of galactosylation 587:611 Regarding main glycoforms, degree of galactosylation was 52% in CT-A produced from F2N78 cells compared to CHO cells (37%). 28054185 8 60 theme CONCLUSIONS 1063:1073 arg1 cells 1081:1085 CONCLUSIONS F2N78 cells 1063:1085 CONCLUSIONS F2N78 cells 1063:1085 CONCLUSIONS F2N78 cells stably produced antibodies with human specific N-glycan. 28054185 7 61 located observed 1038:1045 arg2 structure 1018:1026 bisecting structure 1008:1026 bisecting structure 1008:1026 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 7 61 located observed 1038:1045 arg2 structure 994:1002 Hybrid structure 987:1002 Hybrid structure 987:1002 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 7 61 located observed 1038:1045 arg1 cells 1056:1060 F2N78 cells 1050:1060 F2N78 cells 1050:1060 Hybrid structure and bisecting structure were only observed in F2N78 cells. 28054185 9 62 theme alternative 1230:1240 arg1 cell 1247:1250 an alternative host cell 1227:1250 an alternative host cell 1227:1250 The novel expression system based on human cells may facilitate the development of an alternative host cell for production of recombinant proteins. 28054185 1 63 theme cell 179:182 arg1 line 184:187 a novel human cell line 165:187 a novel human cell line 165:187 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 1 63 theme cell 179:182 arg1 F2N78 190:194 F2N78 190:194 F2N78 190:194 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 2 64 theme physicochemical 353:367 arg1 properties 369:378 physicochemical properties 353:378 physicochemical properties of antibodies produced from F2N78 and CHO cells 353:426 RESULTS The culture performance and physicochemical properties of antibodies produced from F2N78 and CHO cells were compared. 28054185 5 65 theme F2N78 818:822 arg1 cells 824:828 F2N78 cells 818:828 F2N78 cells 818:828 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 1 66 theme growth 207:212 arg1 performance 214:224 growth performance 207:224 growth performance 207:224 OBJECTIVES To evaluate the characteristics of a novel human cell line, F2N78, including growth performance, physicochemical properties, and biological activity via direct comparison with CHO cells. 28054185 5 67 theme α-2,6-linked 707:718 arg1 residues 767:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues 707:774 For sialic acid forms, α-2,6-linked sialic acid and N-acetylneuraminic acid (NANA) residues were observed in antibodies produced from F2N78 cells. 28054185 3 68 contain contained 501:509 arg1 antibodies 464:473 antibodies 464:473 antibodies produced from F2N78 cells 464:499 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 3 68 contain contained 501:509 arg2 variants 535:542 a greater acidic charge variants 511:542 a greater acidic charge variants 511:542 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28054185 0 69 dep Characterization 0:15 arg1 comparison 61:70 a comparison 59:70 a comparison of culture performances and protein qualities 59:116 Characterization of human hybrid cell line, F2N78, through a comparison of culture performances and protein qualities. 28054185 3 70 theme greater 513:519 arg1 variants 535:542 a greater acidic charge variants 511:542 a greater acidic charge variants 511:542 For charge variants, antibodies produced from F2N78 cells contained a greater acidic charge variants than CHO cells. 28905148 2 0 theme glycan 489:494 arg1 assignments 506:516 glycan structure assignments 489:516 glycan structure assignments 489:516 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 3 1 theme Ribonuclease 703:714 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 1 theme Ribonuclease 703:714 arg1 B 716:716 Ribonuclease B 703:716 Ribonuclease B (RNase B) 703:726 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 1 theme Ribonuclease 703:714 arg1 B 725:725 RNase B 719:725 RNase B 719:725 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 5 2 theme fragmentation 1188:1200 arg1 spectra 1202:1208 supporting fragmentation spectra 1177:1208 supporting fragmentation spectra 1177:1208 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 5 2 theme fragmentation 1188:1200 arg1 products 1245:1252 exoglycosidase digestion products 1220:1252 exoglycosidase digestion products 1220:1252 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 2 3 dep evaluate 288:295 arg1 provide 354:360 provide 354:360 provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily 354:528 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 4 4 theme tetra-antennary 1045:1059 arg1 classes 1061:1067 sialylated tetra-antennary classes 1034:1067 sialylated tetra-antennary classes 1034:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 2 5 theme separation 401:410 arg1 properties 412:421 chromatographic separation properties 385:421 chromatographic separation properties 385:421 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 3 6 theme α1-acid 681:687 arg1 glycoprotein 689:700 α1-acid glycoprotein 681:700 α1-acid glycoprotein 681:700 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 6 theme α1-acid 681:687 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 5 7 theme N-glycans 1316:1324 arg1 properties 1302:1311 the elution properties 1290:1311 the elution properties of N-glycans by PGC-LC 1290:1334 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 1 8 theme isobaric 252:259 arg1 structures 268:277 isomeric and isobaric glycan structures 239:277 isomeric and isobaric glycan structures 239:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 4 9 theme retention 931:939 arg1 library 941:947 a N-glycan PGC retention library 916:947 a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 916:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 3 10 from profile 753:759 arg1 PGC-LC-MS 790:798 capillary PGC-LC-MS 780:798 capillary PGC-LC-MS 780:798 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 4 11 theme glycan 810:815 arg1 structures 817:826 Over 100 glycan structures 801:826 Over 100 glycan structures 801:826 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 3 12 theme reported 550:557 arg1 structures 568:577 previously reported N-glycan structures 539:577 previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 539:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 2 13 from behaviour 309:317 arg1 PGC-LC 335:340 PGC-LC 335:340 PGC-LC 335:340 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 1 14 theme glycan 261:266 arg1 structures 268:277 isomeric and isobaric glycan structures 239:277 isomeric and isobaric glycan structures 239:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 4 15 theme PGC 927:929 arg1 library 941:947 a N-glycan PGC retention library 916:947 a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 916:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 3 16 theme RNase 719:723 arg1 B 716:716 Ribonuclease B 703:716 Ribonuclease B (RNase B) 703:726 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 16 theme RNase 719:723 arg1 B 725:725 RNase B 719:725 RNase B 719:725 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 5 17 theme elution 1294:1300 arg1 properties 1302:1311 the elution properties 1290:1311 the elution properties of N-glycans by PGC-LC 1290:1334 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 3 18 theme N-glycan 559:566 arg1 structures 568:577 previously reported N-glycan structures 539:577 previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 539:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 1 19 theme structures 268:277 arg1 mixtures 227:234 complex mixtures 219:234 complex mixtures of isomeric and isobaric glycan structures 219:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 6 20 theme glycoproteins 1516:1528 arg1 analysis 1431:1438 the detailed analysis 1418:1438 the detailed analysis of the glycosylation of both purified recombinant 1418:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 6 20 theme glycoproteins 1516:1528 arg1 mixtures 1503:1510 complex mixtures 1495:1510 complex mixtures of, glycoproteins using established workflows 1495:1556 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 1 21 theme powerful 159:166 arg1 chromatography 111:124 Porous graphitised carbon-liquid chromatography 78:124 Porous graphitised carbon-liquid chromatography (PGC-LC) 78:133 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 21 theme powerful 159:166 arg1 technique 168:176 a powerful technique 157:176 a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures 157:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 5 22 theme detailed 1268:1275 arg1 knowledge 1277:1285 detailed knowledge 1268:1285 detailed knowledge on the elution properties of N-glycans by PGC-LC 1268:1334 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 6 23 theme detailed 1422:1429 arg1 analysis 1431:1438 the detailed analysis 1418:1438 the detailed analysis of the glycosylation of both purified recombinant 1418:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 6 24 theme recombinant 1478:1488 arg1 glycosylation 1447:1459 the glycosylation 1443:1459 the glycosylation of both purified recombinant 1443:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 1 25 theme Porous 78:83 arg1 PGC-LC 127:132 PGC-LC 127:132 PGC-LC 127:132 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 25 theme Porous 78:83 arg1 chromatography 111:124 Porous graphitised carbon-liquid chromatography 78:124 Porous graphitised carbon-liquid chromatography (PGC-LC) 78:133 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 25 theme Porous 78:83 arg1 technique 168:176 a powerful technique 157:176 a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures 157:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 3 26 attach released 579:586 arg1 A 659:659 Immunoglobulin A 644:659 Immunoglobulin A (IgA) 644:665 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 ovalbumin 740:748 ovalbumin 740:748 ovalbumin 740:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 glycoprotein 689:700 α1-acid glycoprotein 681:700 α1-acid glycoprotein 681:700 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg2 structures 568:577 previously reported N-glycan structures 539:577 previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 539:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 fetuin 729:734 fetuin 729:734 fetuin 729:734 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 lactoferrin 668:678 lactoferrin 668:678 lactoferrin 668:678 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 26 attach released 579:586 arg1 B 716:716 Ribonuclease B 703:716 Ribonuclease B (RNase B) 703:726 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 2 27 theme chromatographic 385:399 arg1 properties 412:421 chromatographic separation properties 385:421 chromatographic separation properties 385:421 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 5 28 theme digestion 1235:1243 arg1 products 1245:1252 exoglycosidase digestion products 1220:1252 exoglycosidase digestion products 1220:1252 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 6 29 theme glycosylation 1447:1459 arg1 analysis 1431:1438 the detailed analysis 1418:1438 the detailed analysis of the glycosylation of both purified recombinant 1418:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 6 29 theme glycosylation 1447:1459 arg1 mixtures 1503:1510 complex mixtures 1495:1510 complex mixtures of, glycoproteins using established workflows 1495:1556 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 1 30 theme graphitised 85:95 arg1 PGC-LC 127:132 PGC-LC 127:132 PGC-LC 127:132 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 30 theme graphitised 85:95 arg1 chromatography 111:124 Porous graphitised carbon-liquid chromatography 78:124 Porous graphitised carbon-liquid chromatography (PGC-LC) 78:133 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 30 theme graphitised 85:95 arg1 technique 168:176 a powerful technique 157:176 a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures 157:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 4 31 theme pauci 1017:1021 arg1 mannose 1023:1029 pauci mannose 1017:1029 pauci mannose 1017:1029 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 1 32 theme carbon-liquid 97:109 arg1 PGC-LC 127:132 PGC-LC 127:132 PGC-LC 127:132 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 32 theme carbon-liquid 97:109 arg1 chromatography 111:124 Porous graphitised carbon-liquid chromatography 78:124 Porous graphitised carbon-liquid chromatography (PGC-LC) 78:133 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 32 theme carbon-liquid 97:109 arg1 technique 168:176 a powerful technique 157:176 a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures 157:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 0 33 theme N-glycan 21:28 arg1 library 40:46 a PGC-LC-MS N-glycan retention library 9:46 a PGC-LC-MS N-glycan retention library 9:46 Building a PGC-LC-MS N-glycan retention library and elution mapping resource. 28905148 5 34 theme exoglycosidase 1220:1233 arg1 products 1245:1252 exoglycosidase digestion products 1220:1252 exoglycosidase digestion products 1220:1252 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 2 35 theme elution 301:307 arg1 behaviour 309:317 the elution behaviour 297:317 the elution behaviour of N-glycans on PGC-LC 297:340 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 2 36 theme mass 438:441 arg1 MS 457:458 MS 457:458 MS 457:458 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 2 36 theme mass 438:441 arg1 spectrometry 443:454 mass spectrometry 438:454 mass spectrometry (MS) fragmentation 438:473 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 4 37 from mannose 1023:1029 arg1 N-glycans 1002:1010 biologically significant N-glycans 977:1010 biologically significant N-glycans from pauci mannose 977:1029 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 4 37 from mannose 1023:1029 arg1 spectrum 965:972 a full spectrum 958:972 a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 958:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 6 38 theme valuable 1383:1390 arg1 platform 1356:1363 this platform 1351:1363 this platform 1351:1363 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 6 38 theme valuable 1383:1390 arg1 resource 1392:1399 a valuable resource 1381:1399 a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows 1381:1556 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 5 39 theme resultant 1074:1082 arg1 library 1098:1104 The resultant PGC retention library 1070:1104 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) 1070:1142 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 4 40 theme significant 990:1000 arg1 N-glycans 1002:1010 biologically significant N-glycans 977:1010 biologically significant N-glycans from pauci mannose 977:1029 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 4 41 theme N-glycan 918:925 arg1 library 941:947 a N-glycan PGC retention library 916:947 a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 916:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 5 42 theme PGC 1084:1086 arg1 library 1098:1104 The resultant PGC retention library 1070:1104 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) 1070:1142 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 3 43 theme Immunoglobulin 620:633 arg1 IgG 638:640 IgG 638:640 IgG 638:640 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 43 theme Immunoglobulin 620:633 arg1 G 635:635 Immunoglobulin G 620:635 Immunoglobulin G (IgG) 620:641 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 4 44 theme sialylated 1034:1043 arg1 classes 1061:1067 sialylated tetra-antennary classes 1034:1067 sialylated tetra-antennary classes 1034:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 3 45 gly glycoproteins 606:618 arg1 A 659:659 Immunoglobulin A 644:659 Immunoglobulin A (IgA) 644:665 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 ovalbumin 740:748 ovalbumin 740:748 ovalbumin 740:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 glycoprotein 689:700 α1-acid glycoprotein 681:700 α1-acid glycoprotein 681:700 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 fetuin 729:734 fetuin 729:734 fetuin 729:734 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 lactoferrin 668:678 lactoferrin 668:678 lactoferrin 668:678 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 45 gly glycoproteins 606:618 arg1 B 716:716 Ribonuclease B 703:716 Ribonuclease B (RNase B) 703:726 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 0 46 theme retention 30:38 arg1 library 40:46 a PGC-LC-MS N-glycan retention library 9:46 a PGC-LC-MS N-glycan retention library 9:46 Building a PGC-LC-MS N-glycan retention library and elution mapping resource. 28905148 6 47 theme purified 1469:1476 arg1 recombinant 1478:1488 both purified recombinant 1464:1488 both purified recombinant 1464:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 3 48 theme Immunoglobulin 644:657 arg1 IgA 662:664 IgA 662:664 IgA 662:664 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 48 theme Immunoglobulin 644:657 arg1 A 659:659 Immunoglobulin A 644:659 Immunoglobulin A (IgA) 644:665 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 48 theme Immunoglobulin 644:657 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 2 49 theme spectrometry 443:454 arg1 fragmentation 461:473 mass spectrometry (MS) fragmentation 438:473 mass spectrometry (MS) fragmentation 438:473 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 5 50 dep http 1108:1111 arg1 //www.glycostore.org/showPgc 1113:1140 //www.glycostore.org/showPgc 1113:1140 http://www.glycostore.org/showPgc 1108:1140 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 0 51 theme elution 52:58 arg1 resource 68:75 elution mapping resource 52:75 elution mapping resource 52:75 Building a PGC-LC-MS N-glycan retention library and elution mapping resource. 28905148 3 52 dep glycoproteins 606:618 arg1 IgG 638:640 IgG 638:640 IgG 638:640 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 52 dep glycoproteins 606:618 arg1 G 635:635 Immunoglobulin G 620:635 Immunoglobulin G (IgG) 620:641 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 4 53 theme targeted 872:879 arg1 digestions 896:905 targeted exoglycosidase digestions 872:905 targeted exoglycosidase digestions 872:905 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 0 54 theme mapping 60:66 arg1 resource 68:75 elution mapping resource 52:75 elution mapping resource 52:75 Building a PGC-LC-MS N-glycan retention library and elution mapping resource. 28905148 3 55 theme purified 597:604 arg1 A 659:659 Immunoglobulin A 644:659 Immunoglobulin A (IgA) 644:665 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 ovalbumin 740:748 ovalbumin 740:748 ovalbumin 740:748 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 glycoprotein 689:700 α1-acid glycoprotein 681:700 α1-acid glycoprotein 681:700 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 fetuin 729:734 fetuin 729:734 fetuin 729:734 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 lactoferrin 668:678 lactoferrin 668:678 lactoferrin 668:678 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 55 theme purified 597:604 arg1 B 716:716 Ribonuclease B 703:716 Ribonuclease B (RNase B) 703:726 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 6 56 theme complex 1495:1501 arg1 mixtures 1503:1510 complex mixtures 1495:1510 complex mixtures of, glycoproteins using established workflows 1495:1556 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 5 57 theme retention 1157:1165 arg1 products 1245:1252 exoglycosidase digestion products 1220:1252 exoglycosidase digestion products 1220:1252 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 5 57 theme retention 1157:1165 arg1 times 1167:1171 retention times 1157:1171 retention times 1157:1171 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 4 58 theme N-glycans 1002:1010 arg1 spectrum 965:972 a full spectrum 958:972 a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 958:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 3 59 gly glycoprotein 689:700 arg1 glycoprotein 689:700 α1-acid glycoprotein 681:700 α1-acid glycoprotein 681:700 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 3 59 gly glycoprotein 689:700 arg1 glycoproteins 606:618 the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin 593:748 glycoproteins 606:618 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 5 60 dep library 1098:1104 arg1 http 1108:1111 http 1108:1111 http://www.glycostore.org/showPgc 1108:1140 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 4 61 gly sialylated 1034:1043 arg1 classes 1061:1067 sialylated tetra-antennary classes 1034:1067 sialylated tetra-antennary classes 1034:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 5 62 theme retention 1088:1096 arg1 library 1098:1104 The resultant PGC retention library 1070:1104 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) 1070:1142 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 4 63 from spectrum 965:972 arg1 mannose 1023:1029 pauci mannose 1017:1029 pauci mannose 1017:1029 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 5 64 from knowledge 1277:1285 arg1 properties 1302:1311 the elution properties 1290:1311 the elution properties of N-glycans by PGC-LC 1290:1334 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 1 65 theme complex 219:225 arg1 mixtures 227:234 complex mixtures 219:234 complex mixtures of isomeric and isobaric glycan structures 219:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 4 66 theme exoglycosidase 881:894 arg1 digestions 896:905 targeted exoglycosidase digestions 872:905 targeted exoglycosidase digestions 872:905 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 6 67 gly glycosylation 1447:1459 arg1 recombinant 1478:1488 both purified recombinant 1464:1488 both purified recombinant 1464:1488 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 2 68 theme N-glycans 322:330 arg1 behaviour 309:317 the elution behaviour 297:317 the elution behaviour of N-glycans on PGC-LC 297:340 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 1 69 theme mixtures 227:234 arg1 characterisation 199:214 characterisation 199:214 characterisation 199:214 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 1 69 theme mixtures 227:234 arg1 analysis 186:193 analysis 186:193 analysis 186:193 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 4 70 theme full 960:963 arg1 spectrum 965:972 a full spectrum 958:972 a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes 958:1067 Over 100 glycan structures were determined by MS/MS, and together with targeted exoglycosidase digestions, created a N-glycan PGC retention library covering a full spectrum of biologically significant N-glycans from pauci mannose to sialylated tetra-antennary classes. 28905148 1 71 dep analysis 186:193 arg1 the 182:184 the 182:184 the 182:184 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 28905148 6 72 theme established 1536:1546 arg1 workflows 1548:1556 established workflows 1536:1556 established workflows 1536:1556 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 3 73 used used 534:537 arg2 We 531:532 We 531:532 We 531:532 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 2 74 theme structure 496:504 arg1 assignments 506:516 glycan structure assignments 489:516 glycan structure assignments 489:516 Here we evaluate the elution behaviour of N-glycans on PGC-LC and thereby provide the potential of using chromatographic separation properties, together with mass spectrometry (MS) fragmentation, to determine glycan structure assignments more easily. 28905148 5 75 theme supporting 1177:1186 arg1 spectra 1202:1208 supporting fragmentation spectra 1177:1208 supporting fragmentation spectra 1177:1208 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 5 75 theme supporting 1177:1186 arg1 products 1245:1252 exoglycosidase digestion products 1220:1252 exoglycosidase digestion products 1220:1252 The resultant PGC retention library ( http://www.glycostore.org/showPgc ) incorporates retention times and supporting fragmentation spectra including exoglycosidase digestion products, and provides detailed knowledge on the elution properties of N-glycans by PGC-LC. 28905148 0 76 theme PGC-LC-MS 11:19 arg1 library 40:46 a PGC-LC-MS N-glycan retention library 9:46 a PGC-LC-MS N-glycan retention library 9:46 Building a PGC-LC-MS N-glycan retention library and elution mapping resource. 28905148 3 77 theme capillary 780:788 arg1 PGC-LC-MS 790:798 capillary PGC-LC-MS 780:798 capillary PGC-LC-MS 780:798 We used previously reported N-glycan structures released from the purified glycoproteins Immunoglobulin G (IgG), Immunoglobulin A (IgA), lactoferrin, α1-acid glycoprotein, Ribonuclease B (RNase B), fetuin and ovalbumin to profile their behaviour on capillary PGC-LC-MS. 28905148 6 78 gly glycoproteins 1516:1528 arg1 glycoproteins 1516:1528 glycoproteins 1516:1528 glycoproteins 1516:1528 Consequently, this platform should serve as a valuable resource for facilitating the detailed analysis of the glycosylation of both purified recombinant, and complex mixtures of, glycoproteins using established workflows. 28905148 1 79 theme isomeric 239:246 arg1 structures 268:277 isomeric and isobaric glycan structures 239:277 isomeric and isobaric glycan structures 239:277 Porous graphitised carbon-liquid chromatography (PGC-LC) has been proven to be a powerful technique for the analysis and characterisation of complex mixtures of isomeric and isobaric glycan structures. 24302562 6 0 theme acid 1183:1186 arg1 derivatives 1188:1198 2-aminobenzoic acid derivatives 1168:1198 2-aminobenzoic acid derivatives (2AA) 1168:1204 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 0 theme acid 1183:1186 arg1 2AA 1201:1203 2AA 1201:1203 2AA 1201:1203 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 1 theme class 1278:1282 arg1 separation 1284:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 4 2 theme quantitative 612:623 arg1 profiling 625:633 quantitative profiling 612:633 quantitative profiling 612:633 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 2 3 theme protein 283:289 arg1 support 257:263 the support 253:263 the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization 253:379 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 8 4 theme specific 1773:1780 arg1 moieties 1782:1789 specific moieties 1773:1789 specific moieties 1773:1789 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 4 theme specific 1773:1780 arg1 sulfation 1819:1827 sulfation 1819:1827 sulfation 1819:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 4 theme specific 1773:1780 arg1 O-acetylation 1801:1813 O-acetylation 1801:1813 O-acetylation 1801:1813 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 4 5 theme structural 677:686 arg1 characterization 688:703 -based structural characterization 670:703 -based structural characterization of sialylated RFP N-glycans 670:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 5 6 theme ion 936:938 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 6 theme ion 936:938 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 2 7 theme stage 316:320 arg1 development 330:340 early stage process development 310:340 early stage process development 310:340 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 1 8 theme protein 176:182 arg1 quality 184:190 protein quality 176:190 protein quality 176:190 Glycosylation is a critical parameter used to evaluate protein quality and consistency. 24302562 6 9 theme anion-exchange 1234:1247 arg1 chromatography 1249:1262 hydrophilic interaction anion-exchange chromatography 1210:1262 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 9 theme anion-exchange 1234:1247 arg1 HIAX 1265:1268 HIAX 1265:1268 HIAX 1265:1268 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 0 10 theme offline 108:114 arg1 MSn 116:118 offline MSn 108:118 offline MSn 108:118 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 5 11 theme online 907:912 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 11 theme online 907:912 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 8 12 theme chromatographic 1548:1562 arg1 dimension 1564:1572 the second chromatographic dimension 1537:1572 the second chromatographic dimension 1537:1572 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 7 13 theme MS/MS 1455:1459 arg1 advantages 1403:1412 the advantages 1399:1412 the advantages of high mass accuracy and data-dependent MS/MS 1399:1459 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 2 14 theme N-linked 209:216 arg1 profiling 225:233 N-linked glycan profiling 209:233 N-linked glycan profiling 209:233 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 5 15 theme static 986:991 arg1 NSI-MS 1044:1049 NSI-MS 1044:1049 NSI-MS(n) 1044:1052 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 15 theme static 986:991 arg1 spectrometry 1030:1041 offline static nanospray ionization-sequential mass spectrometry 978:1041 offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)) 978:1053 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 8 16 theme orthogonal 1585:1594 arg1 eluants 1661:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants 1585:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation 1585:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 16 theme orthogonal 1585:1594 arg1 investigations 1716:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 7 17 theme mass 1422:1425 arg1 accuracy 1427:1434 high mass accuracy 1417:1434 high mass accuracy 1417:1434 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 4 18 theme RFP 719:721 arg1 N-glycans 723:731 sialylated RFP N-glycans 708:731 sialylated RFP N-glycans 708:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 7 19 theme fractions 1334:1342 arg1 characterization 1304:1319 Further characterization 1296:1319 Further characterization of HIAX peak fractions 1296:1342 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 6 20 theme routine 1060:1066 arg1 use 1068:1070 routine use 1060:1070 routine use 1060:1070 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 7 21 theme HIAX 1324:1327 arg1 fractions 1334:1342 HIAX peak fractions 1324:1342 HIAX peak fractions 1324:1342 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 3 22 theme Sialylated 382:391 arg1 glycans 393:399 Sialylated glycans 382:399 Sialylated glycans 382:399 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 8 23 theme ion 1600:1602 arg1 spectra 1610:1616 ion MS/MS spectra 1600:1616 ion MS/MS spectra 1600:1616 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 0 24 theme online 82:87 arg1 MS/MS 98:102 online orbitrap MS/MS 82:102 online orbitrap MS/MS 82:102 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 8 25 theme spectra 1610:1616 arg1 eluants 1661:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants 1585:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation 1585:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 25 theme spectra 1610:1616 arg1 investigations 1716:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 6 26 dep employs 1122:1128 arg1 established 1130:1140 established 1130:1140 employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1122:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 7 27 theme high 1417:1420 arg1 accuracy 1427:1434 high mass accuracy 1417:1434 high mass accuracy 1417:1434 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 3 28 theme serum 412:416 arg1 half-life 418:426 the serum half-life 408:426 the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins 408:553 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 8 29 theme buffer-free 1622:1632 arg1 chromatography 1641:1654 buffer-free liquid chromatography 1622:1654 buffer-free liquid chromatography 1622:1654 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 5 30 theme detailed 796:803 arg1 characterization 816:831 detailed structural characterization 796:831 detailed structural characterization 796:831 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 31 theme ionization-sequential 1003:1023 arg1 NSI-MS 1044:1049 NSI-MS 1044:1049 NSI-MS(n) 1044:1052 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 31 theme ionization-sequential 1003:1023 arg1 spectrometry 1030:1041 offline static nanospray ionization-sequential mass spectrometry 978:1041 offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)) 978:1053 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 32 theme routine 761:767 arg1 studies 783:789 routine comparability studies 761:789 routine comparability studies 761:789 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 8 33 theme chromatography 1641:1654 arg1 eluants 1661:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants 1585:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation 1585:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 33 theme chromatography 1641:1654 arg1 investigations 1716:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 3 34 theme proteins 546:553 arg1 production 525:534 the production 521:534 the production of fusion proteins 521:553 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 3 35 theme fusion 443:448 arg1 RFPs 460:463 RFPs 460:463 RFPs 460:463 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 3 35 theme fusion 443:448 arg1 proteins 450:457 receptor-Fc fusion proteins 431:457 receptor-Fc fusion proteins (RFPs) 431:464 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 6 36 theme chromatography 1097:1110 arg1 profiling 1112:1120 high-performance liquid chromatography profiling 1073:1120 high-performance liquid chromatography profiling 1073:1120 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 8 37 theme additional 1475:1484 arg1 characterization 1486:1501 additional characterization 1475:1501 additional characterization 1475:1501 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 0 38 theme sialylated 34:43 arg1 N-glycans 45:53 sialylated N-glycans 34:53 sialylated N-glycans 34:53 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 5 39 theme integrated 837:846 arg1 workflow 848:855 an integrated workflow 834:855 an integrated workflow 834:855 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 6 40 theme liquid 1090:1095 arg1 chromatography 1097:1110 high-performance liquid chromatography 1073:1110 high-performance liquid chromatography profiling 1073:1120 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 5 41 theme structural 805:814 arg1 characterization 816:831 detailed structural characterization 796:831 detailed structural characterization 796:831 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 2 42 theme product 355:361 arg1 commercialization 363:379 drug product commercialization 350:379 drug product commercialization 350:379 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 7 43 theme ion 1370:1372 arg1 MS/MS 1383:1387 online (-) ion orbitrap MS/MS 1359:1387 online (-) ion orbitrap MS/MS 1359:1387 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 8 44 theme graphitized 1515:1525 arg1 carbon 1527:1532 porous graphitized carbon 1508:1532 porous graphitized carbon in the second chromatographic dimension 1508:1572 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 6 45 theme derivatives 1188:1198 arg1 separation 1284:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 45 theme derivatives 1188:1198 arg1 detection 1155:1163 fluorescence detection 1142:1163 fluorescence detection of 2-aminobenzoic acid derivatives (2AA) 1142:1204 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 4 46 theme in-depth 639:646 arg1 MS 667:668 MS 667:668 MS 667:668 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 4 46 theme in-depth 639:646 arg1 spectrometry 653:664 in-depth mass spectrometry 639:664 in-depth mass spectrometry (MS) 639:669 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 6 47 theme 2-aminobenzoic 1168:1181 arg1 derivatives 1188:1198 2-aminobenzoic acid derivatives 1168:1198 2-aminobenzoic acid derivatives (2AA) 1168:1204 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 47 theme 2-aminobenzoic 1168:1181 arg1 2AA 1201:1203 2AA 1201:1203 2AA 1201:1203 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 8 48 theme low-abundance 1747:1759 arg1 sulfation 1819:1827 sulfation 1819:1827 sulfation 1819:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 48 theme low-abundance 1747:1759 arg1 O-acetylation 1801:1813 O-acetylation 1801:1813 O-acetylation 1801:1813 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 48 theme low-abundance 1747:1759 arg1 species 1761:1767 low-abundance species 1747:1767 low-abundance species 1747:1767 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 6 49 theme charge 1271:1276 arg1 separation 1284:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 2 50 theme biotherapeutic 268:281 arg1 protein 283:289 biotherapeutic protein 268:289 biotherapeutic protein manufacturing from early stage process development through drug product commercialization 268:379 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 5 51 theme negative 927:934 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 51 theme negative 927:934 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 4 52 theme -based 670:675 arg1 characterization 688:703 -based structural characterization 670:703 -based structural characterization of sialylated RFP N-glycans 670:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 2 53 theme process 322:328 arg1 development 330:340 early stage process development 310:340 early stage process development 310:340 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 5 54 theme tandem 940:945 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 54 theme tandem 940:945 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 6 55 theme chromatography 1249:1262 arg1 separation 1284:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 2 56 theme early 310:314 arg1 development 330:340 early stage process development 310:340 early stage process development 310:340 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 8 57 from carbon 1527:1532 arg1 dimension 1564:1572 the second chromatographic dimension 1537:1572 the second chromatographic dimension 1537:1572 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 58 theme second 1541:1546 arg1 dimension 1564:1572 the second chromatographic dimension 1537:1572 the second chromatographic dimension 1537:1572 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 6 59 theme interaction 1222:1232 arg1 chromatography 1249:1262 hydrophilic interaction anion-exchange chromatography 1210:1262 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 59 theme interaction 1222:1232 arg1 HIAX 1265:1268 HIAX 1265:1268 HIAX 1265:1268 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 5 60 theme mass 947:950 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 60 theme mass 947:950 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 61 theme positive 914:921 arg1 MS/MS 966:970 MS/MS 966:970 MS/MS 966:970 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 61 theme positive 914:921 arg1 spectrometry 952:963 online positive and negative ion tandem mass spectrometry 907:963 online positive and negative ion tandem mass spectrometry (MS/MS) 907:971 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 0 62 theme orbitrap 89:96 arg1 MS/MS 98:102 online orbitrap MS/MS 82:102 online orbitrap MS/MS 82:102 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 4 63 gly sialylated 708:717 arg1 N-glycans 723:731 sialylated RFP N-glycans 708:731 sialylated RFP N-glycans 708:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 2 64 theme glycan 218:223 arg1 profiling 225:233 N-linked glycan profiling 209:233 N-linked glycan profiling 209:233 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 0 65 gly sialylated 34:43 arg1 N-glycans 45:53 sialylated N-glycans 34:53 sialylated N-glycans 34:53 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 5 66 theme offline 978:984 arg1 NSI-MS 1044:1049 NSI-MS 1044:1049 NSI-MS(n) 1044:1052 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 66 theme offline 978:984 arg1 spectrometry 1030:1041 offline static nanospray ionization-sequential mass spectrometry 978:1041 offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)) 978:1053 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 6 67 theme high-performance 1073:1088 arg1 chromatography 1097:1110 high-performance liquid chromatography 1073:1110 high-performance liquid chromatography profiling 1073:1120 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 7 68 theme data-dependent 1440:1453 arg1 MS/MS 1455:1459 data-dependent MS/MS 1440:1459 data-dependent MS/MS 1440:1459 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 2 69 link N-linked 209:216 arg1 profiling 225:233 N-linked glycan profiling 209:233 N-linked glycan profiling 209:233 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 7 70 theme peak 1329:1332 arg1 fractions 1334:1342 HIAX peak fractions 1324:1342 HIAX peak fractions 1324:1342 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 4 71 dep profiling 625:633 arg1 characterization 688:703 -based structural characterization 670:703 -based structural characterization of sialylated RFP N-glycans 670:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 7 72 theme accuracy 1427:1434 arg1 advantages 1403:1412 the advantages 1399:1412 the advantages of high mass accuracy and data-dependent MS/MS 1399:1459 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 4 73 theme sialylated 708:717 arg1 N-glycans 723:731 sialylated RFP N-glycans 708:731 sialylated RFP N-glycans 708:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 5 74 theme comparability 769:781 arg1 studies 783:789 routine comparability studies 761:789 routine comparability studies 761:789 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 1 75 theme critical 140:147 arg1 Glycosylation 121:133 Glycosylation 121:133 Glycosylation 121:133 Glycosylation is a critical parameter used to evaluate protein quality and consistency. 24302562 1 75 theme critical 140:147 arg1 parameter 149:157 a critical parameter 138:157 a critical parameter used to evaluate protein quality and consistency 138:206 Glycosylation is a critical parameter used to evaluate protein quality and consistency. 24302562 8 76 theme MS/MS 1604:1608 arg1 spectra 1610:1616 ion MS/MS spectra 1600:1616 ion MS/MS spectra 1600:1616 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 4 77 theme N-glycans 723:731 arg1 characterization 688:703 -based structural characterization 670:703 -based structural characterization of sialylated RFP N-glycans 670:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 7 78 theme Further 1296:1302 arg1 characterization 1304:1319 Further characterization 1296:1319 Further characterization of HIAX peak fractions 1296:1342 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 8 79 theme liquid 1634:1639 arg1 chromatography 1641:1654 buffer-free liquid chromatography 1622:1654 buffer-free liquid chromatography 1622:1654 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 5 80 theme nanospray 993:1001 arg1 NSI-MS 1044:1049 NSI-MS 1044:1049 NSI-MS(n) 1044:1052 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 80 theme nanospray 993:1001 arg1 spectrometry 1030:1041 offline static nanospray ionization-sequential mass spectrometry 978:1041 offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)) 978:1053 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 6 81 theme fluorescence 1142:1153 arg1 detection 1155:1163 fluorescence detection 1142:1163 fluorescence detection of 2-aminobenzoic acid derivatives (2AA) 1142:1204 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 8 82 theme peak 1656:1659 arg1 eluants 1661:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants 1585:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation 1585:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 82 theme peak 1656:1659 arg1 investigations 1716:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 3 83 dep proteins 450:457 arg1 making 467:472 making 467:472 making their quality and consistency a concern during the production of fusion proteins 467:553 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 5 84 theme fluorescence 883:894 arg1 detection 896:904 fluorescence detection 883:904 fluorescence detection 883:904 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 85 theme mass 1025:1028 arg1 NSI-MS 1044:1049 NSI-MS 1044:1049 NSI-MS(n) 1044:1052 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 5 85 theme mass 1025:1028 arg1 spectrometry 1030:1041 offline static nanospray ionization-sequential mass spectrometry 978:1041 offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)) 978:1053 Aiming to efficiently link routine comparability studies with detailed structural characterization, an integrated workflow was implemented employing fluorescence detection, online positive and negative ion tandem mass spectrometry (MS/MS), and offline static nanospray ionization-sequential mass spectrometry (NSI-MS(n)). 24302562 4 86 theme analytical 577:586 arg1 approach 588:595 an analytical approach 574:595 an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans 574:731 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 0 87 theme N-glycans 45:53 arg1 characterization 14:29 characterization 14:29 characterization 14:29 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 0 87 theme N-glycans 45:53 arg1 Profiling 0:8 Profiling 0:8 Profiling 0:8 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 7 88 theme orbitrap 1374:1381 arg1 MS/MS 1383:1387 online (-) ion orbitrap MS/MS 1359:1387 online (-) ion orbitrap MS/MS 1359:1387 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 3 89 theme proteins 450:457 arg1 half-life 418:426 the serum half-life 408:426 the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins 408:553 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 8 90 theme optimum 1678:1684 arg1 eluants 1661:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants 1585:1667 orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation 1585:1827 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 90 theme optimum 1678:1684 arg1 investigations 1716:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 optimum for offline (+)/(-) NSI-MS(n) investigations 1678:1729 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 6 91 theme hydrophilic 1210:1220 arg1 chromatography 1249:1262 hydrophilic interaction anion-exchange chromatography 1210:1262 hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation 1210:1293 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 6 91 theme hydrophilic 1210:1220 arg1 HIAX 1265:1268 HIAX 1265:1268 HIAX 1265:1268 For routine use, high-performance liquid chromatography profiling employs established fluorescence detection of 2-aminobenzoic acid derivatives (2AA) and hydrophilic interaction anion-exchange chromatography (HIAX) charge class separation. 24302562 3 92 theme receptor-Fc 431:441 arg1 RFPs 460:463 RFPs 460:463 RFPs 460:463 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 3 92 theme receptor-Fc 431:441 arg1 proteins 450:457 receptor-Fc fusion proteins 431:457 receptor-Fc fusion proteins (RFPs) 431:464 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 8 93 theme offline 1690:1696 arg1 n 1713:1713 n 1713:1713 n 1713:1713 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 8 93 theme offline 1690:1696 arg1 NSI-MS 1706:1711 offline (+)/(-) NSI-MS 1690:1711 offline (+)/(-) NSI-MS(n) 1690:1714 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 2 94 theme drug 350:353 arg1 commercialization 363:379 drug product commercialization 350:379 drug product commercialization 350:379 N-linked glycan profiling is fundamental to the support of biotherapeutic protein manufacturing from early stage process development through drug product commercialization. 24302562 3 95 dep making 467:472 arg1 consistency 492:502 consistency 492:502 consistency 492:502 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 3 95 dep making 467:472 arg1 quality 480:486 quality 480:486 quality 480:486 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 24302562 8 96 theme porous 1508:1513 arg1 carbon 1527:1532 porous graphitized carbon 1508:1532 porous graphitized carbon in the second chromatographic dimension 1508:1572 As required, additional characterization uses porous graphitized carbon in the second chromatographic dimension to provide orthogonal (+) ion MS/MS spectra and buffer-free liquid chromatography peak eluants that are optimum for offline (+)/(-) NSI-MS(n) investigations to characterize low-abundance species and specific moieties including O-acetylation and sulfation. 24302562 0 97 with 2D-HPLC 58:64 arg1 MS/MS 98:102 online orbitrap MS/MS 82:102 online orbitrap MS/MS 82:102 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 0 97 with 2D-HPLC 58:64 arg1 MSn 116:118 offline MSn 108:118 offline MSn 108:118 Profiling and characterization of sialylated N-glycans by 2D-HPLC (HIAX/PGC) with online orbitrap MS/MS and offline MSn. 24302562 7 98 theme online 1359:1364 arg1 MS/MS 1383:1387 online (-) ion orbitrap MS/MS 1359:1387 online (-) ion orbitrap MS/MS 1359:1387 Further characterization of HIAX peak fractions is achieved by online (-) ion orbitrap MS/MS, offering the advantages of high mass accuracy and data-dependent MS/MS. 24302562 4 99 theme mass 648:651 arg1 MS 667:668 MS 667:668 MS 667:668 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 4 99 theme mass 648:651 arg1 spectrometry 653:664 in-depth mass spectrometry 639:664 in-depth mass spectrometry (MS) 639:669 Here, we describe an analytical approach providing both quantitative profiling and in-depth mass spectrometry (MS)-based structural characterization of sialylated RFP N-glycans. 24302562 3 100 theme fusion 539:544 arg1 proteins 546:553 fusion proteins 539:553 fusion proteins 539:553 Sialylated glycans impact the serum half-life of receptor-Fc fusion proteins (RFPs), making their quality and consistency a concern during the production of fusion proteins. 28434916 0 0 theme Specific 80:87 arg1 Glycans 95:101 Specific Viral Glycans 80:101 Specific Viral Glycans 80:101 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 7 1 dep glycan 1138:1143 arg1 shield 1145:1150 shield 1145:1150 shield 1145:1150 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 1 2 contain have 119:122 arg1 lectins 111:117 Select lectins 104:117 Select lectins 104:117 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 1 2 contain have 119:122 arg2 properties 144:153 powerful anti-viral properties 124:153 powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus 124:237 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 3 3 theme BanLec 486:491 arg1 monomer 493:499 each BanLec monomer 481:499 each BanLec monomer that were previously considered separate carbohydrate recognition domains 481:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 5 4 from stoichiometry 725:737 arg1 solution 742:749 solution 742:749 solution 742:749 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 3 5 theme recognition 555:565 arg1 domains 567:573 separate carbohydrate recognition domains 533:573 separate carbohydrate recognition domains 533:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 0 6 theme Viral 89:93 arg1 Glycans 95:101 Specific Viral Glycans 80:101 Specific Viral Glycans 80:101 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 3 7 theme separate 533:540 arg1 domains 567:573 separate carbohydrate recognition domains 533:573 separate carbohydrate recognition domains 533:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 1 8 theme anti-viral 133:142 arg1 properties 144:153 powerful anti-viral properties 124:153 powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus 124:237 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 7 9 theme HIV 1155:1157 arg1 glycan 1138:1143 the dense glycan shield 1128:1150 the dense glycan shield of HIV 1128:1157 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 4 10 theme glycan 631:636 arg1 binding 638:644 high-affinity glycan binding 617:644 high-affinity glycan binding 617:644 We show that both sites are required for high-affinity glycan binding and virus neutralization. 28434916 3 11 theme bidentate 416:424 arg1 interactions 426:437 bidentate interactions 416:437 bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains 416:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 0 12 theme Tetrameric 4:13 arg1 BanLec 28:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 6 13 theme individual 910:919 arg1 glycans 921:927 individual glycans 910:927 individual glycans 910:927 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 2 14 theme potent 295:300 arg1 lectins 302:308 the most potent lectins 286:308 the most potent lectins 286:308 Here, we reveal the mechanism by which one of the most potent lectins, BanLec, achieves its inhibition. 28434916 3 15 theme high-mannose 391:402 arg1 glycans 404:410 high-mannose glycans 391:410 high-mannose glycans 391:410 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 3 16 theme glycans 404:410 arg1 subset 381:386 a subset 379:386 a subset of high-mannose glycans 379:410 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 0 17 theme Lectin 21:26 arg1 BanLec 28:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 6 18 theme avidity 1011:1017 arg1 effects 1019:1025 enhanced avidity effects 1002:1025 enhanced avidity effects 1002:1025 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 7 19 theme dense 1132:1136 arg1 glycan 1138:1143 the dense glycan shield 1128:1150 the dense glycan shield of HIV 1128:1157 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 3 20 theme carbohydrate 542:553 arg1 domains 567:573 separate carbohydrate recognition domains 533:573 separate carbohydrate recognition domains 533:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 1 21 theme dense 206:210 arg1 glycan 212:217 the dense glycan shield 202:224 the dense glycan shield on the virus 202:237 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 0 22 theme Plant 15:19 arg1 BanLec 28:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec 0:33 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 6 23 theme enhanced 1002:1009 arg1 effects 1019:1025 enhanced avidity effects 1002:1025 enhanced avidity effects 1002:1025 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 1 24 theme Select 104:109 arg1 lectins 111:117 Select lectins 104:117 Select lectins 104:117 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 4 25 theme high-affinity 617:629 arg1 binding 638:644 high-affinity glycan binding 617:644 high-affinity glycan binding 617:644 We show that both sites are required for high-affinity glycan binding and virus neutralization. 28434916 5 26 theme tetrameric 714:723 arg1 stoichiometry 725:737 a tetrameric stoichiometry 712:737 a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes 712:843 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 6 27 theme bidentate 889:897 arg1 binding 899:905 bidentate binding 889:905 bidentate binding to individual glycans 889:927 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 6 28 theme viral 973:977 arg1 neutralization 979:992 viral neutralization 973:992 viral neutralization 973:992 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 7 29 theme lectin 1098:1103 arg1 therapeutics 1105:1116 successful lectin therapeutics 1087:1116 successful lectin therapeutics targeting the dense glycan shield of HIV 1087:1157 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 3 30 theme present 470:476 arg1 sites 464:468 the two binding sites 448:468 the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains 448:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 5 31 theme glycosylated 819:830 arg1 spikes 838:843 glycosylated viral spikes 819:843 glycosylated viral spikes 819:843 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 1 32 from glycan 212:217 arg1 virus 233:237 the virus 229:237 the virus 229:237 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 1 33 theme powerful 124:131 arg1 properties 144:153 powerful anti-viral properties 124:153 powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus 124:237 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 7 34 theme successful 1087:1096 arg1 therapeutics 1105:1116 successful lectin therapeutics 1087:1116 successful lectin therapeutics targeting the dense glycan shield of HIV 1087:1157 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 7 35 theme structural 1034:1043 arg1 insights 1045:1052 These structural insights 1028:1052 These structural insights 1028:1052 These structural insights will prove useful in engineering successful lectin therapeutics targeting the dense glycan shield of HIV. 28434916 2 36 theme lectins 302:308 arg1 BanLec 311:316 BanLec 311:316 BanLec 311:316 Here, we reveal the mechanism by which one of the most potent lectins, BanLec, achieves its inhibition. 28434916 2 36 theme lectins 302:308 arg1 lectins 302:308 the most potent lectins 286:308 the most potent lectins 286:308 Here, we reveal the mechanism by which one of the most potent lectins, BanLec, achieves its inhibition. 28434916 2 36 theme lectins 302:308 arg1 one 279:281 one 279:281 one 279:281 Here, we reveal the mechanism by which one of the most potent lectins, BanLec, achieves its inhibition. 28434916 3 37 theme binding 456:462 arg1 sites 464:468 the two binding sites 448:468 the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains 448:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 5 38 theme glycan-binding 763:776 arg1 sites 778:782 the glycan-binding sites 759:782 the glycan-binding sites 759:782 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 5 38 theme glycan-binding 763:776 arg1 positioned 788:797 positioned 788:797 positioned 788:797 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 6 39 theme tetrameric 850:859 arg1 architecture 861:872 The tetrameric architecture 846:872 The tetrameric architecture 846:872 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 5 40 gly glycosylated 819:830 arg1 spikes 838:843 glycosylated viral spikes 819:843 glycosylated viral spikes 819:843 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 28434916 3 41 attach present 470:476 arg1 monomer 493:499 each BanLec monomer 481:499 each BanLec monomer that were previously considered separate carbohydrate recognition domains 481:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 3 41 attach present 470:476 arg2 sites 464:468 the two binding sites 448:468 the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains 448:573 We identify that BanLec recognizes a subset of high-mannose glycans via bidentate interactions spanning the two binding sites present on each BanLec monomer that were previously considered separate carbohydrate recognition domains. 28434916 4 42 theme virus 650:654 arg1 neutralization 656:669 virus neutralization 650:669 virus neutralization 650:669 We show that both sites are required for high-affinity glycan binding and virus neutralization. 28434916 0 43 theme Bidentate 59:67 arg1 Binding 69:75 Bidentate Binding 59:75 Bidentate Binding to Specific Viral Glycans 59:101 The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. 28434916 6 44 theme multivalency 949:960 arg1 layers 939:944 layers 939:944 layers of multivalency that drive viral neutralization through enhanced avidity effects 939:1025 The tetrameric architecture, together with bidentate binding to individual glycans, leads to layers of multivalency that drive viral neutralization through enhanced avidity effects. 28434916 1 45 dep glycan 212:217 arg1 shield 219:224 shield 219:224 shield 219:224 Select lectins have powerful anti-viral properties that effectively neutralize HIV-1 by targeting the dense glycan shield on the virus. 28434916 5 46 theme viral 832:836 arg1 spikes 838:843 glycosylated viral spikes 819:843 glycosylated viral spikes 819:843 Unexpectedly we find that BanLec adopts a tetrameric stoichiometry in solution whereby the glycan-binding sites are positioned to optimally target glycosylated viral spikes. 27311591 0 0 theme IgG 102:104 arg1 Samples 106:112 Human IgG Samples 96:112 Human IgG Samples 96:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 6 1 theme enzyme-linked 1244:1256 arg1 assays 1273:1278 enzyme-linked lectin binding assays 1244:1278 enzyme-linked lectin binding assays (ELLBAs) 1244:1287 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 1 theme enzyme-linked 1244:1256 arg1 ELLBAs 1281:1286 ELLBAs 1281:1286 ELLBAs 1281:1286 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 4 2 theme bioreceptive 694:705 arg1 layer 707:711 a bioreceptive layer 692:711 a bioreceptive layer 692:711 The process of building a bioreceptive layer was optimized and characterized using a diverse range of techniques. 27311591 7 3 theme glycosylation 1498:1510 arg1 changes 1512:1518 glycosylation changes 1498:1518 glycosylation changes in human IgGs 1498:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 0 4 theme Human 96:100 arg1 Samples 106:112 Human IgG Samples 96:112 Human IgG Samples 96:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 6 5 theme immunoglobulins 1065:1079 arg1 IgGs 1084:1087 immunoglobulins G (IgGs) 1065:1088 immunoglobulins G (IgGs) isolated from serum samples 1065:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 2 6 theme nonspecific 516:526 arg1 interactions 528:539 nonspecific interactions 516:539 nonspecific interactions 516:539 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 5 7 theme serum 985:989 arg1 samples 991:997 human serum samples 979:997 human serum samples 979:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 6 8 theme other 1150:1154 arg1 methods 1179:1185 two other standard bioanalytical methods 1146:1185 two other standard bioanalytical methods employing lectins 1146:1203 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 7 9 theme sensitive 1485:1493 arg1 procedure 1475:1483 a new diagnostic procedure 1458:1483 a new diagnostic procedure sensitive to glycosylation changes in human IgGs 1458:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 0 10 theme Samples 106:112 arg1 Analyses 84:91 Impedimetric Glycomic Analyses 62:91 Impedimetric Glycomic Analyses of Human IgG Samples 62:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 6 11 theme G 1081:1081 arg1 IgGs 1084:1087 immunoglobulins G (IgGs) 1065:1088 immunoglobulins G (IgGs) isolated from serum samples 1065:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 7 12 theme RA 1354:1355 arg1 score 1374:1378 RA disease activity score 28 1354:1381 RA disease activity score 28 1354:1381 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 12 theme RA 1354:1355 arg1 index 1347:1351 the DAS28 index 1337:1351 the DAS28 index (RA disease activity score 28) 1337:1382 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 0 13 theme Array 120:124 arg1 Format 126:131 an Array Format 117:131 an Array Format 117:131 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 7 14 theme impedimetric 1401:1412 arg1 assays 1414:1419 impedimetric assays 1401:1419 impedimetric assays 1401:1419 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 15 theme disease 1357:1363 arg1 score 1374:1378 RA disease activity score 28 1354:1381 RA disease activity score 28 1354:1381 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 15 theme disease 1357:1363 arg1 index 1347:1351 the DAS28 index 1337:1351 the DAS28 index (RA disease activity score 28) 1337:1382 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 16 theme activity 1365:1372 arg1 score 1374:1378 RA disease activity score 28 1354:1381 RA disease activity score 28 1354:1381 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 16 theme activity 1365:1372 arg1 index 1347:1351 the DAS28 index 1337:1351 the DAS28 index (RA disease activity score 28) 1337:1382 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 2 17 theme mixed 364:368 arg1 SAM 396:398 SAM 396:398 SAM 396:398 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 17 theme mixed 364:368 arg1 monolayer 385:393 a mixed self-assembled monolayer 362:393 a mixed self-assembled monolayer (SAM) 362:399 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 0 18 theme Impedimetric 62:73 arg1 Analyses 84:91 Impedimetric Glycomic Analyses 62:91 Impedimetric Glycomic Analyses of Human IgG Samples 62:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 4 19 theme techniques 770:779 arg1 range 761:765 a diverse range 751:765 a diverse range of techniques 751:779 The process of building a bioreceptive layer was optimized and characterized using a diverse range of techniques. 27311591 6 20 link enzyme-linked 1244:1256 arg1 assays 1273:1278 enzyme-linked lectin binding assays 1244:1278 enzyme-linked lectin binding assays (ELLBAs) 1244:1287 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 20 link enzyme-linked 1244:1256 arg1 ELLBAs 1281:1286 ELLBAs 1281:1286 ELLBAs 1281:1286 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 2 21 theme self-assembled 370:383 arg1 SAM 396:398 SAM 396:398 SAM 396:398 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 21 theme self-assembled 370:383 arg1 monolayer 385:393 a mixed self-assembled monolayer 362:393 a mixed self-assembled monolayer (SAM) 362:399 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 0 22 theme Zwitterion-Based 6:21 arg1 Interface 48:56 Zwitterion-Based Self-Assembled Monolayer Interface 6:56 Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples 6:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 6 23 theme lectin 1215:1220 arg1 microarrays 1222:1232 lectin microarrays 1215:1232 lectin microarrays (MAs) 1215:1238 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 23 theme lectin 1215:1220 arg1 MAs 1235:1237 MAs 1235:1237 MAs 1235:1237 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 5 24 theme moderate 928:935 arg1 level 937:941 a moderate level 926:941 a moderate level of multiplexing 926:957 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 6 25 theme impedimetric 1040:1051 arg1 analysis 1053:1060 the impedimetric analysis 1036:1060 the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples 1036:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 1 26 theme changes 188:194 arg1 detection 175:183 the detection 171:183 the detection of changes in the glycan structure of antibodies isolated from human serum 171:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 7 27 theme RA 1543:1544 arg1 progression 1546:1556 RA progression 1543:1556 RA progression 1543:1556 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 5 28 theme level 937:941 arg1 achievement 911:921 the achievement 907:921 the achievement of a moderate level of multiplexing for the analysis of human serum samples 907:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 0 29 theme Monolayer 38:46 arg1 Interface 48:56 Zwitterion-Based Self-Assembled Monolayer Interface 6:56 Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples 6:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 6 30 theme bioanalytical 1165:1177 arg1 methods 1179:1185 two other standard bioanalytical methods 1146:1185 two other standard bioanalytical methods employing lectins 1146:1203 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 4 31 theme diverse 753:759 arg1 range 761:765 a diverse range 751:765 a diverse range of techniques 751:779 The process of building a bioreceptive layer was optimized and characterized using a diverse range of techniques. 27311591 3 32 theme covalent 595:602 arg1 immobilization 604:617 the covalent immobilization 591:617 the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I) 591:665 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 0 33 theme Self-Assembled 23:36 arg1 Interface 48:56 Zwitterion-Based Self-Assembled Monolayer Interface 6:56 Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples 6:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 7 34 theme diagnostic 1464:1473 arg1 procedure 1475:1483 a new diagnostic procedure 1458:1483 a new diagnostic procedure sensitive to glycosylation changes in human IgGs 1458:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 5 35 theme multiplexing 946:957 arg1 level 937:941 a moderate level 926:941 a moderate level of multiplexing 926:957 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 1 36 theme glycan 203:208 arg1 structure 210:218 the glycan structure 199:218 the glycan structure of antibodies isolated from human serum 199:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 5 37 theme Impedimetric 782:793 arg1 assays 795:800 Impedimetric assays 782:800 Impedimetric assays 782:800 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 7 38 theme procedure 1475:1483 arg1 development 1443:1453 the development 1439:1453 the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs 1439:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 38 theme procedure 1475:1483 arg1 progression 1546:1556 RA progression 1543:1556 RA progression 1543:1556 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 1 39 attach isolated 234:241 arg2 antibodies 223:232 antibodies 223:232 antibodies isolated from human serum 223:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 1 39 attach isolated 234:241 arg1 serum 254:258 human serum 248:258 human serum 248:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 5 40 theme gold 848:851 arg1 step 895:898 an important step 882:898 an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples 882:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 5 40 theme gold 848:851 arg1 electrodes 861:870 eight gold working electrodes 842:870 eight gold working electrodes 842:870 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 2 41 theme zwitterionic 447:458 arg1 sulfobetaine 492:503 sulfobetaine 492:503 sulfobetaine 492:503 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 41 theme zwitterionic 447:458 arg1 carboxybetaine 473:486 carboxybetaine 473:486 carboxybetaine 473:486 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 41 theme zwitterionic 447:458 arg1 derivatives 460:470 two different thiolated zwitterionic derivatives 423:470 two different thiolated zwitterionic derivatives 423:470 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 1 42 theme rheumatoid 303:312 arg1 arthritis 314:322 rheumatoid arthritis 303:322 rheumatoid arthritis (RA) 303:327 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 1 42 theme rheumatoid 303:312 arg1 RA 325:326 RA 325:326 RA 325:326 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 7 43 from changes 1512:1518 arg1 IgGs 1529:1532 human IgGs 1523:1532 human IgGs 1523:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 3 44 theme agglutinin 646:655 arg1 immobilization 604:617 the covalent immobilization 591:617 the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I) 591:665 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 7 45 theme DAS28 1341:1345 arg1 score 1374:1378 RA disease activity score 28 1354:1381 RA disease activity score 28 1354:1381 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 7 45 theme DAS28 1341:1345 arg1 index 1347:1351 the DAS28 index 1337:1351 the DAS28 index (RA disease activity score 28) 1337:1382 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 1 46 from structure 210:218 arg1 detection 175:183 the detection 171:183 the detection of changes in the glycan structure of antibodies isolated from human serum 171:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 5 47 theme working 853:859 arg1 step 895:898 an important step 882:898 an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples 882:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 5 47 theme working 853:859 arg1 electrodes 861:870 eight gold working electrodes 842:870 eight gold working electrodes 842:870 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 0 48 theme Glycomic 75:82 arg1 Analyses 84:91 Impedimetric Glycomic Analyses 62:91 Impedimetric Glycomic Analyses of Human IgG Samples 62:112 Mixed Zwitterion-Based Self-Assembled Monolayer Interface for Impedimetric Glycomic Analyses of Human IgG Samples in an Array Format. 27311591 1 49 theme arthritis 314:322 arg1 progression 288:298 the progression 284:298 the progression of rheumatoid arthritis (RA) 284:327 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 7 50 theme impedimetric 1294:1305 arg1 results 1307:1313 The impedimetric results 1290:1313 The impedimetric results 1290:1313 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 6 51 theme IgGs 1084:1087 arg1 analysis 1053:1060 the impedimetric analysis 1036:1060 the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples 1036:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 52 theme serum 1104:1108 arg1 samples 1110:1116 serum samples 1104:1116 serum samples 1104:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 2 53 theme different 427:435 arg1 sulfobetaine 492:503 sulfobetaine 492:503 sulfobetaine 492:503 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 53 theme different 427:435 arg1 carboxybetaine 473:486 carboxybetaine 473:486 carboxybetaine 473:486 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 53 theme different 427:435 arg1 derivatives 460:470 two different thiolated zwitterionic derivatives 423:470 two different thiolated zwitterionic derivatives 423:470 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 6 54 theme standard 1156:1163 arg1 methods 1179:1185 two other standard bioanalytical methods 1146:1185 two other standard bioanalytical methods employing lectins 1146:1203 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 55 theme binding 1265:1271 arg1 assays 1273:1278 enzyme-linked lectin binding assays 1244:1278 enzyme-linked lectin binding assays (ELLBAs) 1244:1287 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 55 theme binding 1265:1271 arg1 ELLBAs 1281:1286 ELLBAs 1281:1286 ELLBAs 1281:1286 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 5 56 theme human 979:983 arg1 samples 991:997 human serum samples 979:997 human serum samples 979:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 3 57 theme Ricinus 629:635 arg1 agglutinin 646:655 lectin Ricinus communis agglutinin I 622:657 lectin Ricinus communis agglutinin I (RCA-I) 622:665 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 3 57 theme Ricinus 629:635 arg1 RCA-I 660:664 RCA-I 660:664 RCA-I 660:664 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 1 58 theme antibodies 223:232 arg1 structure 210:218 the glycan structure 199:218 the glycan structure of antibodies isolated from human serum 199:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 6 59 theme lectin 1258:1263 arg1 assays 1273:1278 enzyme-linked lectin binding assays 1244:1278 enzyme-linked lectin binding assays (ELLBAs) 1244:1287 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 59 theme lectin 1258:1263 arg1 ELLBAs 1281:1286 ELLBAs 1281:1286 ELLBAs 1281:1286 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 3 60 theme communis 637:644 arg1 agglutinin 646:655 lectin Ricinus communis agglutinin I 622:657 lectin Ricinus communis agglutinin I (RCA-I) 622:665 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 3 60 theme communis 637:644 arg1 RCA-I 660:664 RCA-I 660:664 RCA-I 660:664 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 1 61 from changes 188:194 arg1 structure 210:218 the glycan structure 199:218 the glycan structure of antibodies isolated from human serum 199:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 1 62 theme impedimetric 137:148 arg1 biosensor 157:165 An impedimetric lectin biosensor 134:165 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum 134:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 6 63 attach isolated 1090:1097 arg1 samples 1110:1116 serum samples 1104:1116 serum samples 1104:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 6 63 attach isolated 1090:1097 arg2 IgGs 1084:1087 immunoglobulins G (IgGs) 1065:1088 immunoglobulins G (IgGs) isolated from serum samples 1065:1116 At the end, the results obtained by the impedimetric analysis of immunoglobulins G (IgGs) isolated from serum samples were compared with those of two other standard bioanalytical methods employing lectins, that is, lectin microarrays (MAs) and enzyme-linked lectin binding assays (ELLBAs). 27311591 5 64 theme samples 991:997 arg1 analysis 967:974 the analysis 963:974 the analysis of human serum samples 963:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 7 65 theme human 1523:1527 arg1 IgGs 1529:1532 human IgGs 1523:1532 human IgGs 1523:1532 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 1 66 theme lectin 150:155 arg1 biosensor 157:165 An impedimetric lectin biosensor 134:165 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum 134:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 7 67 used used 1430:1433 arg2 assays 1414:1419 impedimetric assays 1401:1419 impedimetric assays 1401:1419 The impedimetric results agreed very well with the DAS28 index (RA disease activity score 28), suggesting that impedimetric assays could be used for the development of a new diagnostic procedure sensitive to glycosylation changes in human IgGs and thus RA progression. 27311591 2 68 theme thiolated 437:445 arg1 sulfobetaine 492:503 sulfobetaine 492:503 sulfobetaine 492:503 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 68 theme thiolated 437:445 arg1 carboxybetaine 473:486 carboxybetaine 473:486 carboxybetaine 473:486 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 2 68 theme thiolated 437:445 arg1 derivatives 460:470 two different thiolated zwitterionic derivatives 423:470 two different thiolated zwitterionic derivatives 423:470 The biosensor was built up from a mixed self-assembled monolayer (SAM) on gold consisting of two different thiolated zwitterionic derivatives, carboxybetaine and sulfobetaine, to resist nonspecific interactions. 27311591 3 69 theme lectin 622:627 arg1 agglutinin 646:655 lectin Ricinus communis agglutinin I 622:657 lectin Ricinus communis agglutinin I (RCA-I) 622:665 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 3 69 theme lectin 622:627 arg1 RCA-I 660:664 RCA-I 660:664 RCA-I 660:664 The carboxyl-terminated one was applied also for the covalent immobilization of lectin Ricinus communis agglutinin I (RCA-I). 27311591 1 70 from detection 175:183 arg1 structure 210:218 the glycan structure 199:218 the glycan structure of antibodies isolated from human serum 199:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 27311591 5 71 theme important 885:893 arg1 step 895:898 an important step 882:898 an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples 882:997 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 5 71 theme important 885:893 arg1 electrodes 861:870 eight gold working electrodes 842:870 eight gold working electrodes 842:870 Impedimetric assays were integrated on a chip consisting of eight gold working electrodes, which is an important step toward the achievement of a moderate level of multiplexing for the analysis of human serum samples. 27311591 1 72 theme human 248:252 arg1 serum 254:258 human serum 248:258 human serum 248:258 An impedimetric lectin biosensor for the detection of changes in the glycan structure of antibodies isolated from human serum is here correlated with the progression of rheumatoid arthritis (RA). 28818784 3 0 theme imprinted 311:319 arg1 cavities 321:328 The eluted imprinted cavities 300:328 The eluted imprinted cavities 300:328 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 5 1 theme better 561:566 arg1 performance 575:585 better sensor performance 561:585 better sensor performance including selectivity and sensitivity 561:623 More complex template structures can lead to better sensor performance including selectivity and sensitivity. 28818784 7 2 theme complex 901:907 arg1 structure 909:917 The complex structure 897:917 The complex structure on the template surface 897:941 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 8 3 theme 103.5 1170:1174 arg1 % 1165:1165 % 1165:1165 % 1165:1165 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 7 4 theme sensor 1116:1121 arg1 capability 1073:1082 the significant anti-interference capability 1039:1082 the significant anti-interference capability of the polysaccharide imprinted sensor 1039:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 6 5 theme imprinted 829:837 arg1 sensor 839:844 the monosaccharide imprinted sensor 810:844 the monosaccharide imprinted sensor 810:844 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 4 6 theme imprinted 476:484 arg1 sensors 486:492 glycosyl imprinted sensors 467:492 glycosyl imprinted sensors 467:492 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 6 7 theme imprinted 658:666 arg1 sensor 668:673 the polysaccharide imprinted sensor 639:673 the polysaccharide imprinted sensor 639:673 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 3 8 theme template 355:362 arg1 glycosyls 364:372 template glycosyls 355:372 template glycosyls 355:372 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 6 9 theme monosaccharide 814:827 arg1 sensor 839:844 the monosaccharide imprinted sensor 810:844 the monosaccharide imprinted sensor 810:844 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 2 10 theme imprinted 282:290 arg1 sensor 292:297 a glycosyl imprinted sensor 271:297 a glycosyl imprinted sensor 271:297 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 6 11 theme polysaccharide 643:656 arg1 sensor 668:673 the polysaccharide imprinted sensor 639:673 the polysaccharide imprinted sensor 639:673 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 7 12 from structure 909:917 arg1 surface 935:941 the template surface 922:941 the template surface 922:941 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 2 13 theme glycosyl 273:280 arg1 sensor 292:297 a glycosyl imprinted sensor 271:297 a glycosyl imprinted sensor 271:297 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 2 14 used used 241:244 arg2 complex 213:219 The glycosylated complex 196:219 The glycosylated complex of glycoprotein 196:235 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 2 14 used used 241:244 arg2 template 249:256 template 249:256 template 249:256 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 7 15 theme template 926:933 arg1 surface 935:941 the template surface 922:941 the template surface 922:941 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 8 16 theme human 1196:1200 arg1 samples 1208:1214 human serum samples 1196:1214 human serum samples 1196:1214 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 7 17 dep provides 943:950 arg1 led 1032:1034 led 1032:1034 led to the significant anti-interference capability of the polysaccharide imprinted sensor 1032:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 5 18 theme complex 521:527 arg1 structures 538:547 More complex template structures 516:547 More complex template structures 516:547 More complex template structures can lead to better sensor performance including selectivity and sensitivity. 28818784 0 19 theme template 31:38 arg1 selection 40:48 an idea template selection 23:48 an idea template selection for glycosyl imprinting 23:72 Does polysaccharide is an idea template selection for glycosyl imprinting? 28818784 0 19 theme template 31:38 arg1 polysaccharide 5:18 polysaccharide 5:18 polysaccharide 5:18 Does polysaccharide is an idea template selection for glycosyl imprinting? 28818784 7 20 theme more 952:955 arg1 possibilities 957:969 more possibilities 952:969 more possibilities 952:969 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 8 21 theme serum 1202:1206 arg1 samples 1208:1214 human serum samples 1196:1214 human serum samples 1196:1214 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 5 22 theme template 529:536 arg1 structures 538:547 More complex template structures 516:547 More complex template structures 516:547 More complex template structures can lead to better sensor performance including selectivity and sensitivity. 28818784 3 23 gly glycoproteins 378:390 arg1 glycoproteins 378:390 glycoproteins 378:390 glycoproteins carrying template glycosyl 378:417 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 8 24 dep % 1165:1165 arg1 to 1167:1168 to 1167:1168 to 1167:1168 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 1 25 theme novel 77:81 arg1 strategy 107:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 1 26 theme carbohydrate 169:180 arg1 19-9 190:193 the carbohydrate antigen 19-9 165:193 the carbohydrate antigen 19-9 165:193 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 3 27 theme template 401:408 arg1 glycosyl 410:417 template glycosyl 401:417 template glycosyl 401:417 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 6 28 theme sensor 839:844 arg1 1-60U/mL 850:857 1-60U/mL 850:857 1-60U/mL 850:857 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 6 28 theme sensor 839:844 arg1 linear 800:805 linear 800:805 linear 800:805 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 4 29 theme template 434:441 arg1 structure 454:462 template saccharide structure 434:462 template saccharide structure 434:462 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 1 30 theme glycoprotein 83:94 arg1 strategy 107:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 1 31 theme antigen 182:188 arg1 19-9 190:193 the carbohydrate antigen 19-9 165:193 the carbohydrate antigen 19-9 165:193 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 7 32 theme polysaccharide 1091:1104 arg1 sensor 1116:1121 the polysaccharide imprinted sensor 1087:1121 the polysaccharide imprinted sensor 1087:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 3 33 contain carrying 392:399 arg1 glycoproteins 378:390 glycoproteins 378:390 glycoproteins carrying template glycosyl 378:417 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 3 33 contain carrying 392:399 arg2 glycosyl 410:417 template glycosyl 401:417 template glycosyl 401:417 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 1 34 theme imprinting 96:105 arg1 strategy 107:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 1 35 theme 19-9 190:193 arg1 detection 152:160 the detection 148:160 the detection of the carbohydrate antigen 19-9 148:193 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 7 36 theme imprinted 1106:1114 arg1 sensor 1116:1121 the polysaccharide imprinted sensor 1087:1121 the polysaccharide imprinted sensor 1087:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 4 37 theme glycosyl 467:474 arg1 sensors 486:492 glycosyl imprinted sensors 467:492 glycosyl imprinted sensors 467:492 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 2 38 gly glycoprotein 224:235 arg1 glycoprotein 224:235 glycoprotein 224:235 glycoprotein 224:235 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 0 39 theme idea 26:29 arg1 selection 40:48 an idea template selection 23:48 an idea template selection for glycosyl imprinting 23:72 Does polysaccharide is an idea template selection for glycosyl imprinting? 28818784 0 39 theme idea 26:29 arg1 polysaccharide 5:18 polysaccharide 5:18 polysaccharide 5:18 Does polysaccharide is an idea template selection for glycosyl imprinting? 28818784 6 40 theme detection 867:875 arg1 0.17U/mL 887:894 0.17U/mL 887:894 0.17U/mL 887:894 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 6 40 theme detection 867:875 arg1 limit 877:881 the detection limit 863:881 the detection limit 863:881 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 2 41 theme glycoprotein 224:235 arg1 complex 213:219 The glycosylated complex 196:219 The glycosylated complex of glycoprotein 196:235 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 2 41 theme glycoprotein 224:235 arg1 template 249:256 template 249:256 template 249:256 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 5 42 theme sensor 568:573 arg1 performance 575:585 better sensor performance 561:585 better sensor performance including selectivity and sensitivity 561:623 More complex template structures can lead to better sensor performance including selectivity and sensitivity. 28818784 8 43 theme imprinted 1254:1262 arg1 sensor 1264:1269 the polysaccharide imprinted sensor 1235:1269 the polysaccharide imprinted sensor 1235:1269 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 1 44 gly glycoprotein 83:94 arg1 glycoprotein 83:94 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy 75:114 A novel glycoprotein imprinting strategy was proposed and was applied to the detection of the carbohydrate antigen 19-9. 28818784 7 45 theme anti-interference 1055:1071 arg1 capability 1073:1082 the significant anti-interference capability 1039:1082 the significant anti-interference capability of the polysaccharide imprinted sensor 1039:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 3 46 theme good 337:340 arg1 affinity 342:349 good affinity 337:349 good affinity for template glycosyls and glycoproteins carrying template glycosyl 337:417 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 8 47 theme polysaccharide 1239:1252 arg1 sensor 1264:1269 the polysaccharide imprinted sensor 1235:1269 the polysaccharide imprinted sensor 1235:1269 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 7 48 theme template 998:1005 arg1 molecules 1007:1015 the template molecules 994:1015 the template molecules 994:1015 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 3 49 theme eluted 304:309 arg1 cavities 321:328 The eluted imprinted cavities 300:328 The eluted imprinted cavities 300:328 The eluted imprinted cavities showed good affinity for template glycosyls and glycoproteins carrying template glycosyl. 28818784 2 50 theme glycosylated 200:211 arg1 complex 213:219 The glycosylated complex 196:219 The glycosylated complex of glycoprotein 196:235 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 2 50 theme glycosylated 200:211 arg1 template 249:256 template 249:256 template 249:256 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 8 51 theme 93.0 1161:1164 arg1 % 1165:1165 % 1165:1165 % 1165:1165 Furthermore, recoveries ranging from 93.0% to 103.5% were achieved when human serum samples were assayed using the polysaccharide imprinted sensor. 28818784 6 52 theme 0.028U/mL 772:780 arg1 limit 763:767 a detection limit 751:767 a detection limit of 0.028U/mL (3δ/K) 751:787 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 2 53 gly glycosylated 200:211 arg1 complex 213:219 The glycosylated complex 196:219 The glycosylated complex of glycoprotein 196:235 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 2 53 gly glycosylated 200:211 arg1 template 249:256 template 249:256 template 249:256 The glycosylated complex of glycoprotein was used as template to construct a glycosyl imprinted sensor. 28818784 7 54 theme significant 1043:1053 arg1 capability 1073:1082 the significant anti-interference capability 1039:1082 the significant anti-interference capability of the polysaccharide imprinted sensor 1039:1121 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 4 55 theme structure 454:462 arg1 effect 424:429 The effect 420:429 The effect of template saccharide structure on glycosyl imprinted sensors 420:492 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 4 56 from effect 424:429 arg1 sensors 486:492 glycosyl imprinted sensors 467:492 glycosyl imprinted sensors 467:492 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 7 57 theme molecules 1007:1015 arg1 recognition 979:989 the recognition 975:989 the recognition of the template molecules 975:1015 The complex structure on the template surface provides more possibilities for the recognition of the template molecules, consequently, led to the significant anti-interference capability of the polysaccharide imprinted sensor. 28818784 6 58 theme linear 693:698 arg1 result 631:636 a result 629:636 a result 629:636 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 6 58 theme linear 693:698 arg1 response 700:707 preeminent linear response 682:707 preeminent linear response to CA19-9 682:717 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 6 59 theme 0.1-5U/mL 735:743 arg1 range 726:730 the range 722:730 the range of 0.1-5U/mL 722:743 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 4 60 theme saccharide 443:452 arg1 structure 454:462 template saccharide structure 434:462 template saccharide structure 434:462 The effect of template saccharide structure on glycosyl imprinted sensors is further discussed. 28818784 6 61 theme preeminent 682:691 arg1 result 631:636 a result 629:636 a result 629:636 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 6 61 theme preeminent 682:691 arg1 response 700:707 preeminent linear response 682:707 preeminent linear response to CA19-9 682:717 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 28818784 0 62 theme glycosyl 54:61 arg1 imprinting 63:72 glycosyl imprinting 54:72 glycosyl imprinting 54:72 Does polysaccharide is an idea template selection for glycosyl imprinting? 28818784 6 63 theme detection 753:761 arg1 limit 763:767 a detection limit 751:767 a detection limit of 0.028U/mL (3δ/K) 751:787 As a result, the polysaccharide imprinted sensor showed preeminent linear response to CA19-9 in the range of 0.1-5U/mL, with a detection limit of 0.028U/mL (3δ/K), while the linear of the monosaccharide imprinted sensor was 1-60U/mL and the detection limit was 0.17U/mL. 27525954 5 0 theme functional 627:636 arg1 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 1 from N-terminal 565:574 arg1 located 550:556 located 550:556 located 550:556 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 2 theme putative 610:617 arg1 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 1 3 theme insect 172:177 arg1 membrane 191:198 the insect peritrophic membrane 168:198 the insect peritrophic membrane 168:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 8 4 theme chitin-binding 1024:1037 arg1 SeCBP66 1047:1053 the novel chitin-binding protein SeCBP66 1014:1053 the novel chitin-binding protein SeCBP66 1014:1053 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 6 5 theme Bac-to-Bac 867:876 arg1 system 889:894 a Bac-to-Bac expression system 865:894 a Bac-to-Bac expression system 865:894 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 5 6 theme glycosylation 719:731 arg1 sites 733:737 O-linked glycosylation sites 710:737 O-linked glycosylation sites 710:737 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 7 theme located 550:556 arg1 acids 544:548 17 amino acids 535:548 17 amino acids located at the N-terminal of SeCBP66 535:585 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 8 8 theme SeCBP66 1047:1053 arg1 study 1005:1009 This study 1000:1009 This study of the novel chitin-binding protein SeCBP66 1000:1053 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 5 9 theme SeCBP66 579:585 arg1 N-terminal 565:574 N-terminal 565:574 N-terminal 565:574 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 2 10 theme immunization 287:298 arg1 screening 300:308 immunization screening 287:308 immunization screening of the cDNA library of Spodoptera exigua 287:349 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 4 11 theme protein 450:456 arg1 weight 436:441 The predicted weight 422:441 The predicted weight of the protein 422:456 The predicted weight of the protein is 64.2 kDa. 27525954 4 11 theme protein 450:456 arg1 kDa 466:468 64.2 kDa 461:468 64.2 kDa 461:468 The predicted weight of the protein is 64.2 kDa. 27525954 5 12 theme Bioinformatic 471:483 arg1 analysis 485:492 Bioinformatic analysis 471:492 Bioinformatic analysis 471:492 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 1 13 theme peritrophic 179:189 arg1 membrane 191:198 the insect peritrophic membrane 168:198 the insect peritrophic membrane 168:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 3 14 theme full 356:359 arg1 bp 387:388 1806 bp 382:388 1806 bp 382:388 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 3 14 theme full 356:359 arg1 length 361:366 The full length 352:366 The full length of secbp66 352:377 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 5 15 theme tandem 603:608 arg1 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 7 16 theme recombinant 941:951 arg1 protein 961:967 the recombinant SeCBP66 protein 937:967 the recombinant SeCBP66 protein 937:967 A chitin binding experiment showed that the recombinant SeCBP66 protein could bind to chitin strongly. 27525954 1 17 theme membrane 191:198 arg1 proteins 131:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 1 17 theme membrane 191:198 arg1 components 154:163 important components 144:163 important components of the insect peritrophic membrane 144:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 6 18 theme expression 878:887 arg1 system 889:894 a Bac-to-Bac expression system 865:894 a Bac-to-Bac expression system 865:894 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 5 19 gly N-glycosylation 680:694 arg2 five 665:668 five 665:668 five 665:668 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 19 gly N-glycosylation 680:694 arg2 sites 696:700 five potential N-glycosylation sites 665:700 five potential N-glycosylation sites 665:700 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 20 contain contained 587:595 arg1 peptide 515:521 a signal peptide 506:521 a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 506:585 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 20 contain contained 587:595 arg2 sites 696:700 five potential N-glycosylation sites 665:700 five potential N-glycosylation sites 665:700 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 20 contain contained 587:595 arg2 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 20 contain contained 587:595 arg2 sites 733:737 O-linked glycosylation sites 710:737 O-linked glycosylation sites 710:737 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 2 21 theme library 322:328 arg1 screening 300:308 immunization screening 287:308 immunization screening of the cDNA library of Spodoptera exigua 287:349 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 3 22 theme amino 409:413 arg1 bp 387:388 1806 bp 382:388 1806 bp 382:388 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 3 22 theme amino 409:413 arg1 acids 415:419 602 amino acids 405:419 602 amino acids 405:419 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 0 23 theme Structural 0:9 arg1 characteristics 27:41 Structural and biochemical characteristics 0:41 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 0:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 5 24 theme Type-II 619:625 arg1 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 8 25 theme novel 1018:1022 arg1 SeCBP66 1047:1053 the novel chitin-binding protein SeCBP66 1014:1053 the novel chitin-binding protein SeCBP66 1014:1053 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 2 26 theme cDNA 317:320 arg1 library 322:328 the cDNA library 313:328 the cDNA library of Spodoptera exigua 313:349 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 4 27 theme predicted 426:434 arg1 weight 436:441 The predicted weight 422:441 The predicted weight of the protein 422:456 The predicted weight of the protein is 64.2 kDa. 27525954 4 27 theme predicted 426:434 arg1 kDa 466:468 64.2 kDa 461:468 64.2 kDa 461:468 The predicted weight of the protein is 64.2 kDa. 27525954 3 28 theme secbp66 371:377 arg1 bp 387:388 1806 bp 382:388 1806 bp 382:388 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 3 28 theme secbp66 371:377 arg1 length 361:366 The full length 352:366 The full length of secbp66 352:377 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 8 29 theme control 1091:1097 arg1 targets 1099:1105 new control targets 1087:1105 new control targets for S. exigua 1087:1119 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 6 30 theme SeCBP66 767:773 arg1 properties 753:762 the properties 749:762 the properties of SeCBP66 749:773 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 0 31 theme biochemical 15:25 arg1 characteristics 27:41 Structural and biochemical characteristics 0:41 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 0:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 8 32 theme new 1087:1089 arg1 targets 1099:1105 new control targets 1087:1105 new control targets for S. exigua 1087:1119 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 5 33 gly glycosylation 719:731 arg2 sites 733:737 O-linked glycosylation sites 710:737 O-linked glycosylation sites 710:737 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 1 34 theme Peritrophic 110:120 arg1 proteins 131:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 1 34 theme Peritrophic 110:120 arg1 components 154:163 important components 144:163 important components of the insect peritrophic membrane 144:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 5 35 theme chitin-binding 638:651 arg1 domains 653:659 seven tandem putative Type-II functional chitin-binding domains 597:659 seven tandem putative Type-II functional chitin-binding domains 597:659 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 0 36 theme chitin-binding 46:59 arg1 protein 61:67 chitin-binding protein SeCBP66 46:75 chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 46:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 0 37 from exigua 93:98 arg1 characteristics 27:41 Structural and biochemical characteristics 0:41 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 0:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 0 37 from exigua 93:98 arg1 protein 61:67 chitin-binding protein SeCBP66 46:75 chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 46:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 3 38 theme 602 405:407 arg1 bp 387:388 1806 bp 382:388 1806 bp 382:388 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 3 38 theme 602 405:407 arg1 acids 415:419 602 amino acids 405:419 602 amino acids 405:419 The full length of secbp66 is 1806 bp, which encodes 602 amino acids. 27525954 5 39 theme O-linked 710:717 arg1 sites 733:737 O-linked glycosylation sites 710:737 O-linked glycosylation sites 710:737 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 2 40 theme chitin-binding 230:243 arg1 protein 245:251 a chitin-binding protein 228:251 a chitin-binding protein 228:251 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 1 41 theme membrane 122:129 arg1 proteins 131:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 1 41 theme membrane 122:129 arg1 components 154:163 important components 144:163 important components of the insect peritrophic membrane 144:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 0 42 theme protein 61:67 arg1 characteristics 27:41 Structural and biochemical characteristics 0:41 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner) 0:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 5 43 theme potential 670:678 arg1 sites 696:700 five potential N-glycosylation sites 665:700 five potential N-glycosylation sites 665:700 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 8 44 theme protein 1039:1045 arg1 SeCBP66 1047:1053 the novel chitin-binding protein SeCBP66 1014:1053 the novel chitin-binding protein SeCBP66 1014:1053 This study of the novel chitin-binding protein SeCBP66 provides a basis for developing new control targets for S. exigua. 27525954 2 45 theme exigua 344:349 arg1 library 322:328 the cDNA library 313:328 the cDNA library of Spodoptera exigua 313:349 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 5 46 theme N-glycosylation 680:694 arg1 sites 696:700 five potential N-glycosylation sites 665:700 five potential N-glycosylation sites 665:700 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 6 47 theme cell 837:840 arg1 line 842:845 the insect cell line BTI-Tn-5B1-4 826:858 the insect cell line BTI-Tn-5B1-4 826:858 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 5 48 theme amino 538:542 arg1 acids 544:548 17 amino acids 535:548 17 amino acids located at the N-terminal of SeCBP66 535:585 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 2 49 theme Spodoptera 333:342 arg1 exigua 344:349 Spodoptera exigua 333:349 Spodoptera exigua 333:349 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 2 50 theme cDNA 209:212 arg1 gene 214:217 A novel cDNA gene 201:217 A novel cDNA gene encoding a chitin-binding protein, named secbp66, 201:267 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 1 51 theme important 144:152 arg1 proteins 131:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins 110:138 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 1 51 theme important 144:152 arg1 components 154:163 important components 144:163 important components of the insect peritrophic membrane 144:198 Peritrophic membrane proteins are important components of the insect peritrophic membrane. 27525954 7 52 theme chitin 899:904 arg1 experiment 914:923 A chitin binding experiment 897:923 A chitin binding experiment 897:923 A chitin binding experiment showed that the recombinant SeCBP66 protein could bind to chitin strongly. 27525954 0 53 theme Spodoptera 82:91 arg1 exigua 93:98 Spodoptera exigua 82:98 Spodoptera exigua (Hübner) 82:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 0 53 theme Spodoptera 82:91 arg1 Hübner 101:106 Hübner 101:106 Hübner 101:106 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 2 54 theme novel 203:207 arg1 gene 214:217 A novel cDNA gene 201:217 A novel cDNA gene encoding a chitin-binding protein, named secbp66, 201:267 A novel cDNA gene encoding a chitin-binding protein, named secbp66, was identified by immunization screening of the cDNA library of Spodoptera exigua. 27525954 6 55 theme insect 830:835 arg1 line 842:845 the insect cell line BTI-Tn-5B1-4 826:858 the insect cell line BTI-Tn-5B1-4 826:858 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 7 56 theme binding 906:912 arg1 experiment 914:923 A chitin binding experiment 897:923 A chitin binding experiment 897:923 A chitin binding experiment showed that the recombinant SeCBP66 protein could bind to chitin strongly. 27525954 6 57 theme recombinant 776:786 arg1 SeCBP66 788:794 recombinant SeCBP66 776:794 recombinant SeCBP66 776:794 To study the properties of SeCBP66, recombinant SeCBP66 was successfully expressed in the insect cell line BTI-Tn-5B1-4 with a Bac-to-Bac expression system. 27525954 7 58 theme SeCBP66 953:959 arg1 protein 961:967 the recombinant SeCBP66 protein 937:967 the recombinant SeCBP66 protein 937:967 A chitin binding experiment showed that the recombinant SeCBP66 protein could bind to chitin strongly. 27525954 0 59 from characteristics 27:41 arg1 exigua 93:98 Spodoptera exigua 82:98 Spodoptera exigua (Hübner) 82:107 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 0 59 from characteristics 27:41 arg1 Hübner 101:106 Hübner 101:106 Hübner 101:106 Structural and biochemical characteristics of chitin-binding protein SeCBP66 from Spodoptera exigua (Hübner). 27525954 5 60 theme signal 508:513 arg1 peptide 515:521 a signal peptide 506:521 a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 506:585 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 27525954 5 61 link O-linked 710:717 arg1 sites 733:737 O-linked glycosylation sites 710:737 O-linked glycosylation sites 710:737 Bioinformatic analysis showed that a signal peptide composed of 17 amino acids located at the N-terminal of SeCBP66 contained seven tandem putative Type-II functional chitin-binding domains and five potential N-glycosylation sites, but no O-linked glycosylation sites. 28486471 9 0 theme biochemical 1636:1646 arg1 data 1660:1663 biochemical and genetic data 1636:1663 biochemical and genetic data reported previously 1636:1683 These profiles are consistent with biochemical and genetic data reported previously. 28486471 10 1 theme glycosylation 1723:1735 arg1 features 1737:1744 glycosylation features 1723:1744 glycosylation features of the cell lines not previously published 1723:1787 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 9 2 theme genetic 1652:1658 arg1 data 1660:1663 biochemical and genetic data 1636:1663 biochemical and genetic data reported previously 1636:1683 These profiles are consistent with biochemical and genetic data reported previously. 28486471 1 3 theme hamster 84:90 arg1 ovary 92:96 Chinese hamster ovary 76:96 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 1 3 theme hamster 84:90 arg1 CHO 99:101 CHO 99:101 CHO 99:101 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 4 4 theme quantitative 664:675 arg1 models 677:682 quantitative models 664:682 quantitative models able to predict glycosylation 664:712 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 6 5 theme CHO 1212:1214 arg1 parent 1185:1190 a wild type parent 1173:1190 a wild type parent 1173:1190 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 6 5 theme CHO 1212:1214 arg1 mutants 1201:1207 nine mutants 1196:1207 nine mutants of CHO 1196:1214 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 0 6 theme Model-based 0:10 arg1 analysis 12:19 Model-based analysis 0:19 Model-based analysis of N-glycosylation in Chinese hamster ovary cells 0:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 7 7 dep 50,605 1350:1355 arg1 to 1347:1348 to 1347:1348 to 1347:1348 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 11 8 theme mutant 2147:2152 arg1 lines 2159:2163 these mutant cell lines 2141:2163 these mutant cell lines 2141:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 4 9 theme able 684:687 arg1 models 677:682 quantitative models 664:682 quantitative models able to predict glycosylation 664:712 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 3 10 theme manipulations 572:584 arg1 effects 553:559 the effects 549:559 the effects of genetic manipulations 549:584 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 1 11 theme proteins 176:183 arg1 manufacturing 134:146 manufacturing 134:146 manufacturing of glycosylated recombinant proteins for production of biotherapeutics 134:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 3 12 theme glycan 592:597 arg1 structures 599:608 glycan structures 592:608 glycan structures of cells 592:617 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 6 13 theme hamster 1129:1135 arg1 ovary 1137:1141 Chinese hamster ovary 1121:1141 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 6 13 theme hamster 1129:1135 arg1 CHO 1144:1146 CHO 1144:1146 CHO 1144:1146 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 5 14 theme same 842:845 arg1 model 847:851 the same model 838:851 the same model used in this study 838:870 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 11 15 theme primary 2076:2082 arg1 loss 2084:2087 primary loss 2076:2087 primary loss 2076:2087 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 7 16 theme glycan 1357:1362 arg1 structures 1364:1373 up to 50,605 glycan structures 1344:1373 up to 50,605 glycan structures 1344:1373 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 1 17 theme biotherapeutics 203:217 arg1 production 189:198 production 189:198 production of biotherapeutics 189:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 11 18 theme function 2114:2121 arg1 gain 2092:2095 gain 2092:2095 gain 2092:2095 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 11 18 theme function 2114:2121 arg1 loss 2084:2087 primary loss 2076:2087 primary loss 2076:2087 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 8 19 theme N-glycan 1431:1438 arg1 spectra 1445:1451 N-glycan mass spectra 1431:1451 N-glycan mass spectra 1431:1451 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 11 20 theme cell 1945:1948 arg1 lines 1950:1954 the CHO cell lines 1937:1954 the CHO cell lines 1937:1954 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 4 21 with deal 749:752 arg1 complexity 763:772 the complexity 759:772 the complexity of glycosylation processing 759:800 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 6 22 theme spectrometry 1269:1280 arg1 data 1282:1285 previously published mass spectrometry data 1243:1285 previously published mass spectrometry data 1243:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 5 23 used used 876:879 arg2 version 827:833 an earlier version 816:833 an earlier version of the same model used in this study 816:870 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 12 24 theme glycoprotein 2360:2371 arg1 products 2373:2380 glycoprotein products 2360:2380 glycoprotein products 2360:2380 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 6 25 theme wild 1175:1178 arg1 parent 1185:1190 a wild type parent 1173:1190 a wild type parent 1173:1190 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 6 26 theme published 1254:1262 arg1 data 1282:1285 previously published mass spectrometry data 1243:1285 previously published mass spectrometry data 1243:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 2 27 theme patterns 256:263 arg1 similarity 224:233 The similarity 220:233 The similarity of its glycosylation patterns to the human versions 220:285 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 8 28 theme detailed 1462:1469 arg1 predictions 1471:1481 detailed predictions 1462:1481 detailed predictions of the glycosylation process 1462:1510 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 7 29 theme mathematical 1316:1327 arg1 model 1329:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 10 30 theme enzyme 1839:1844 arg1 activities 1846:1855 glycosylation enzyme activities 1825:1855 glycosylation enzyme activities than just those resulting directly from gene mutations 1825:1910 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 1 31 theme standard 121:128 arg1 cell 104:107 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 1 31 theme standard 121:128 arg1 gold 116:119 the gold 112:119 the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics 112:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 8 32 theme glycosylation 1490:1502 arg1 process 1504:1510 the glycosylation process 1486:1510 the glycosylation process 1486:1510 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 10 33 from changes 1814:1820 arg1 activities 1846:1855 glycosylation enzyme activities 1825:1855 glycosylation enzyme activities than just those resulting directly from gene mutations 1825:1910 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 12 34 theme glycoform 2294:2302 arg1 distributions 2304:2316 glycoform distributions 2294:2316 glycoform distributions 2294:2316 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 4 35 theme processing 791:800 arg1 complexity 763:772 the complexity 759:772 the complexity of glycosylation processing 759:800 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 12 36 contain have 2212:2215 arg2 potential 2221:2229 the potential 2217:2229 the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products 2217:2380 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 12 36 contain have 2212:2215 arg1 models 2179:2184 Quantitative models 2166:2184 Quantitative models of CHO cell glycosylation 2166:2210 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 5 37 from changes 915:921 arg1 activities 933:942 enzyme activities 926:942 enzyme activities 926:942 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 1 38 theme Chinese 76:82 arg1 ovary 92:96 Chinese hamster ovary 76:96 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 1 38 theme Chinese 76:82 arg1 CHO 99:101 CHO 99:101 CHO 99:101 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 6 39 theme model 1053:1057 arg1 version 1037:1043 an updated version 1026:1043 an updated version of this model 1026:1057 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 8 40 theme activity 1520:1527 arg1 profiles 1529:1536 enzyme activity profiles 1513:1536 enzyme activity profiles 1513:1536 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 2 41 dep line 320:323 arg1 properties 351:360 favorable pharmacokinetic properties 325:360 favorable pharmacokinetic properties 325:360 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 5 42 theme earlier 819:825 arg1 version 827:833 an earlier version 816:833 an earlier version of the same model used in this study 816:870 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 1 43 theme ovary 92:96 arg1 cell 104:107 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell 72:107 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 1 43 theme ovary 92:96 arg1 gold 116:119 the gold 112:119 the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics 112:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 2 44 theme glycosylation 242:254 arg1 patterns 256:263 its glycosylation patterns 238:263 its glycosylation patterns 238:263 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 3 45 theme action 476:481 arg1 product 451:457 the product 447:457 the product of the concerted action of intracellular enzymes 447:506 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 3 45 theme action 476:481 arg1 structures 432:441 glycan structures 425:441 glycan structures 425:441 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 10 46 theme lines 1758:1762 arg1 features 1737:1744 glycosylation features 1723:1744 glycosylation features of the cell lines not previously published 1723:1787 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 3 47 dep predict 528:534 arg1 alter 586:590 alter 586:590 alter glycan structures of cells and therapeutic properties 586:644 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 6 48 theme updated 1029:1035 arg1 version 1037:1043 an updated version 1026:1043 an updated version of this model 1026:1057 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 3 49 theme glycan 425:430 arg1 product 451:457 the product 447:457 the product of the concerted action of intracellular enzymes 447:506 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 3 49 theme glycan 425:430 arg1 structures 432:441 glycan structures 425:441 glycan structures 425:441 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 8 50 theme glycosylation 1551:1563 arg1 profiles 1565:1572 complete glycosylation profiles 1542:1572 complete glycosylation profiles of each of the cell lines 1542:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 12 51 theme glycosylation 2198:2210 arg1 models 2179:2184 Quantitative models 2166:2184 Quantitative models of CHO cell glycosylation 2166:2210 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 11 52 theme CHO 1941:1943 arg1 lines 1950:1954 the CHO cell lines 1937:1954 the CHO cell lines 1937:1954 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 8 53 theme enzyme 1390:1395 arg1 activities 1397:1406 the enzyme activities 1386:1406 the enzyme activities in this model to match N-glycan mass spectra 1386:1451 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 2 54 theme pharmacokinetic 335:349 arg1 properties 351:360 favorable pharmacokinetic properties 325:360 favorable pharmacokinetic properties 325:360 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 0 55 theme hamster 51:57 arg1 cells 65:69 Chinese hamster ovary cells 43:69 Chinese hamster ovary cells 43:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 2 56 theme line 320:323 arg1 products 298:305 the products 294:305 the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses 294:414 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 10 57 theme cell 1753:1756 arg1 lines 1758:1762 the cell lines 1749:1762 the cell lines not previously published 1749:1787 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 5 58 theme glycan 983:988 arg1 structure 990:998 glycan structure 983:998 glycan structure 983:998 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 8 59 theme cell 1589:1592 arg1 lines 1594:1598 the cell lines 1585:1598 the cell lines 1585:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 9 60 with consistent 1620:1629 arg1 data 1660:1663 biochemical and genetic data 1636:1663 biochemical and genetic data reported previously 1636:1683 These profiles are consistent with biochemical and genetic data reported previously. 28486471 10 61 theme gene 1897:1900 arg1 mutations 1902:1910 gene mutations 1897:1910 gene mutations 1897:1910 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 12 62 theme CHO 2189:2191 arg1 glycosylation 2198:2210 CHO cell glycosylation 2189:2210 CHO cell glycosylation 2189:2210 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 2 63 theme likelihood 372:381 arg1 products 298:305 the products 294:305 the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses 294:414 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 1 64 theme glycosylated 151:162 arg1 proteins 176:183 glycosylated recombinant proteins 151:183 glycosylated recombinant proteins for production of biotherapeutics 151:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 6 65 theme comprehensive 1072:1084 arg1 analysis 1086:1093 a comprehensive analysis 1070:1093 a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1070:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 1 66 gly glycosylated 151:162 arg1 proteins 176:183 glycosylated recombinant proteins 151:183 glycosylated recombinant proteins for production of biotherapeutics 151:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 5 67 from change 973:978 arg1 structure 990:998 glycan structure 983:998 glycan structure 983:998 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 6 68 theme cell 1149:1152 arg1 lines 1154:1158 ten Chinese hamster ovary (CHO) cell lines 1117:1158 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 2 69 theme human 272:276 arg1 versions 278:285 the human versions 268:285 the human versions 268:285 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 3 70 theme genetic 564:570 arg1 manipulations 572:584 genetic manipulations 564:584 genetic manipulations 564:584 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 1 71 theme recombinant 164:174 arg1 proteins 176:183 glycosylated recombinant proteins 151:183 glycosylated recombinant proteins for production of biotherapeutics 151:217 The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. 28486471 8 72 theme process 1504:1510 arg1 profiles 1565:1572 complete glycosylation profiles 1542:1572 complete glycosylation profiles of each of the cell lines 1542:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 72 theme process 1504:1510 arg1 profiles 1529:1536 enzyme activity profiles 1513:1536 enzyme activity profiles 1513:1536 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 72 theme process 1504:1510 arg1 predictions 1471:1481 detailed predictions 1462:1481 detailed predictions of the glycosylation process 1462:1510 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 11 73 theme cell 2154:2157 arg1 lines 2159:2163 these mutant cell lines 2141:2163 these mutant cell lines 2141:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 10 74 theme model-based 1690:1700 arg1 results 1702:1708 The model-based results 1686:1708 The model-based results 1686:1708 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 6 75 theme ovary 1137:1141 arg1 lines 1154:1158 ten Chinese hamster ovary (CHO) cell lines 1117:1158 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 11 76 theme side 2056:2059 arg1 effects 2061:2067 side effects 2056:2067 side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines 2056:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 12 77 gly glycoprotein 2360:2371 arg1 glycoprotein 2360:2371 glycoprotein products 2360:2380 glycoprotein products 2360:2380 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 11 78 theme gain 2092:2095 arg1 effects 2061:2067 side effects 2056:2067 side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines 2056:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 6 79 theme Chinese 1121:1127 arg1 ovary 1137:1141 Chinese hamster ovary 1121:1141 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 6 79 theme Chinese 1121:1127 arg1 CHO 1144:1146 CHO 1144:1146 CHO 1144:1146 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 10 80 theme glycosylation 1825:1837 arg1 activities 1846:1855 glycosylation enzyme activities 1825:1855 glycosylation enzyme activities than just those resulting directly from gene mutations 1825:1910 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 8 81 from activities 1397:1406 arg1 model 1416:1420 this model to match N-glycan mass spectra 1411:1451 this model to match N-glycan mass spectra 1411:1451 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 5 82 theme model 847:851 arg1 version 827:833 an earlier version 816:833 an earlier version of the same model used in this study 816:870 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 11 83 theme enzyme 2026:2031 arg1 activities 2033:2042 glycosylation enzyme activities 2012:2042 glycosylation enzyme activities 2012:2042 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 8 84 theme mass 1440:1443 arg1 spectra 1445:1451 N-glycan mass spectra 1431:1451 N-glycan mass spectra 1431:1451 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 6 85 theme data 1282:1285 arg1 interpretation 1225:1238 interpretation 1225:1238 interpretation of previously published mass spectrometry data 1225:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 11 86 theme regulatory 1964:1973 arg1 mechanisms 1975:1984 regulatory mechanisms 1964:1984 regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines 1964:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 12 87 theme products 2373:2380 arg1 performance 2345:2355 the therapeutic performance 2329:2355 the therapeutic performance of glycoprotein products 2329:2380 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 6 88 theme type 1180:1183 arg1 parent 1185:1190 a wild type parent 1173:1190 a wild type parent 1173:1190 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 6 89 theme mass 1264:1267 arg1 data 1282:1285 previously published mass spectrometry data 1243:1285 previously published mass spectrometry data 1243:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 3 90 theme therapeutic 623:633 arg1 properties 635:644 therapeutic properties 623:644 therapeutic properties 623:644 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 11 91 theme glycosylation 2100:2112 arg1 function 2114:2121 glycosylation function 2100:2121 glycosylation function 2100:2121 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 4 92 theme promising 730:738 arg1 tools 740:744 promising tools 730:744 promising tools to deal with the complexity of glycosylation processing 730:800 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 3 93 theme intracellular 486:498 arg1 enzymes 500:506 intracellular enzymes 486:506 intracellular enzymes 486:506 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 3 94 theme cells 613:617 arg1 structures 599:608 glycan structures 592:608 glycan structures of cells 592:617 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 3 94 theme cells 613:617 arg1 properties 635:644 therapeutic properties 623:644 therapeutic properties 623:644 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 11 95 theme glycosylation 2012:2024 arg1 activities 2033:2042 glycosylation enzyme activities 2012:2042 glycosylation enzyme activities 2012:2042 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 12 96 theme therapeutic 2333:2343 arg1 performance 2345:2355 the therapeutic performance 2329:2355 the therapeutic performance of glycoprotein products 2329:2380 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 0 97 from analysis 12:19 arg1 cells 65:69 Chinese hamster ovary cells 43:69 Chinese hamster ovary cells 43:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 11 98 contain possess 1956:1962 arg1 lines 1950:1954 the CHO cell lines 1937:1954 the CHO cell lines 1937:1954 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 11 98 contain possess 1956:1962 arg2 mechanisms 1975:1984 regulatory mechanisms 1964:1984 regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines 1964:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 8 99 theme enzyme 1513:1518 arg1 profiles 1529:1536 enzyme activity profiles 1513:1536 enzyme activity profiles 1513:1536 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 12 100 theme glycoengineering 2250:2265 arg1 manipulations 2267:2279 glycoengineering manipulations 2250:2279 glycoengineering manipulations 2250:2279 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 2 101 theme cell 315:318 arg1 line 320:323 this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses 310:414 line 320:323 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 5 102 theme desired 965:971 arg1 change 973:978 a desired change 963:978 a desired change in glycan structure 963:998 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 7 103 theme updated 1292:1298 arg1 model 1329:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 4 104 theme glycosylation 777:789 arg1 processing 791:800 glycosylation processing 777:800 glycosylation processing 777:800 For that reason, quantitative models able to predict glycosylation have emerged as promising tools to deal with the complexity of glycosylation processing. 28486471 6 105 from lines 1154:1158 arg1 analysis 1086:1093 a comprehensive analysis 1070:1093 a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1070:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 2 106 theme immunogenic 394:404 arg1 responses 406:414 immunogenic responses 394:414 immunogenic responses 394:414 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 5 107 theme enzyme 926:931 arg1 activities 933:942 enzyme activities 926:942 enzyme activities 926:942 For example, an earlier version of the same model used in this study was used by others to successfully predict changes in enzyme activities that could produce a desired change in glycan structure. 28486471 8 108 theme complete 1542:1549 arg1 profiles 1565:1572 complete glycosylation profiles 1542:1572 complete glycosylation profiles of each of the cell lines 1542:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 6 109 gly N-glycosylation 1098:1112 arg1 lines 1154:1158 ten Chinese hamster ovary (CHO) cell lines 1117:1158 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 0 110 theme N-glycosylation 24:38 arg1 analysis 12:19 Model-based analysis 0:19 Model-based analysis of N-glycosylation in Chinese hamster ovary cells 0:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 8 111 gly glycosylation 1551:1563 arg1 each 1577:1580 each 1577:1580 each 1577:1580 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 111 gly glycosylation 1551:1563 arg1 process 1504:1510 the glycosylation process 1486:1510 the glycosylation process 1486:1510 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 111 gly glycosylation 1551:1563 arg1 lines 1594:1598 the cell lines 1585:1598 the cell lines 1585:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 0 112 theme Chinese 43:49 arg1 cells 65:69 Chinese hamster ovary cells 43:69 Chinese hamster ovary cells 43:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 10 113 theme complex 1806:1812 arg1 changes 1814:1820 more complex changes 1801:1820 more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations 1801:1910 The model-based results also predict glycosylation features of the cell lines not previously published, indicating more complex changes in glycosylation enzyme activities than just those resulting directly from gene mutations. 28486471 8 114 theme each 1577:1580 arg1 profiles 1565:1572 complete glycosylation profiles 1542:1572 complete glycosylation profiles of each of the cell lines 1542:1598 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 114 theme each 1577:1580 arg1 profiles 1529:1536 enzyme activity profiles 1513:1536 enzyme activity profiles 1513:1536 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 8 114 theme each 1577:1580 arg1 predictions 1471:1481 detailed predictions 1462:1481 detailed predictions of the glycosylation process 1462:1510 Adjusting the enzyme activities in this model to match N-glycan mass spectra produces detailed predictions of the glycosylation process, enzyme activity profiles and complete glycosylation profiles of each of the cell lines. 28486471 12 115 theme cell 2193:2196 arg1 glycosylation 2198:2210 CHO cell glycosylation 2189:2210 CHO cell glycosylation 2189:2210 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 6 116 from N-glycosylation 1098:1112 arg1 lines 1154:1158 ten Chinese hamster ovary (CHO) cell lines 1117:1158 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 3 117 theme concerted 466:474 arg1 action 476:481 the concerted action 462:481 the concerted action of intracellular enzymes 462:506 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 0 118 theme ovary 59:63 arg1 cells 65:69 Chinese hamster ovary cells 43:69 Chinese hamster ovary cells 43:69 Model-based analysis of N-glycosylation in Chinese hamster ovary cells. 28486471 11 119 theme loss 2084:2087 arg1 effects 2061:2067 side effects 2056:2067 side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines 2056:2163 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 2 120 theme favorable 325:333 arg1 properties 351:360 favorable pharmacokinetic properties 325:360 favorable pharmacokinetic properties 325:360 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 6 121 theme N-glycosylation 1098:1112 arg1 analysis 1086:1093 a comprehensive analysis 1070:1093 a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1070:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 12 122 theme Quantitative 2166:2177 arg1 models 2179:2184 Quantitative models 2166:2184 Quantitative models of CHO cell glycosylation 2166:2210 Quantitative models of CHO cell glycosylation have the potential for predicting how glycoengineering manipulations might affect glycoform distributions to improve the therapeutic performance of glycoprotein products. 28486471 3 123 theme enzymes 500:506 arg1 action 476:481 the concerted action 462:481 the concerted action of intracellular enzymes 462:506 Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties. 28486471 7 124 theme N-glycosylation 1300:1314 arg1 model 1329:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 11 125 dep loss 2084:2087 arg1 the 2072:2074 the 2072:2074 the 2072:2074 The model predicts that the CHO cell lines possess regulatory mechanisms that allow them to adjust glycosylation enzyme activities to mitigate side effects of the primary loss or gain of glycosylation function known to exist in these mutant cell lines. 28486471 6 126 from analysis 1086:1093 arg1 lines 1154:1158 ten Chinese hamster ovary (CHO) cell lines 1117:1158 ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data 1117:1285 In this study we utilize an updated version of this model to provide a comprehensive analysis of N-glycosylation in ten Chinese hamster ovary (CHO) cell lines that include a wild type parent and nine mutants of CHO, through interpretation of previously published mass spectrometry data. 28486471 2 127 theme lower 366:370 arg1 likelihood 372:381 lower likelihood 366:381 lower likelihood of causing immunogenic responses 366:414 The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. 28486471 7 128 contain contains 1335:1342 arg2 structures 1364:1373 up to 50,605 glycan structures 1344:1373 up to 50,605 glycan structures 1344:1373 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 28486471 7 128 contain contains 1335:1342 arg1 model 1329:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model 1288:1333 The updated N-glycosylation mathematical model contains up to 50,605 glycan structures. 24910992 0 0 theme hedgehog 84:91 arg1 degradation 46:56 ER-associated degradation 32:56 ER-associated degradation of non-glycosylated sonic hedgehog 32:91 EDEM2 and OS-9 are required for ER-associated degradation of non-glycosylated sonic hedgehog. 24910992 4 1 theme delivery 496:503 arg1 mechanism 505:513 a similar recognition and delivery mechanism 470:513 mechanism 505:513 Whether a similar recognition and delivery mechanism exists in mammalian cells is unknown. 24910992 5 2 gly nonglycosylated 827:841 arg1 N278A 850:854 its nonglycosylated mutant N278A 823:854 its nonglycosylated mutant N278A 823:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 9 3 with interaction 1230:1240 arg1 variant 1293:1299 a non-glycosylated SHH variant 1270:1299 a non-glycosylated SHH variant 1270:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 0 4 theme sonic 78:82 arg1 hedgehog 84:91 non-glycosylated sonic hedgehog 61:91 non-glycosylated sonic hedgehog 61:91 EDEM2 and OS-9 are required for ER-associated degradation of non-glycosylated sonic hedgehog. 24910992 9 5 theme Robust 1223:1228 arg1 interaction 1230:1240 Robust interaction 1223:1240 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant 1223:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 5 6 theme lectins 653:659 arg1 function 596:603 function 596:603 function 596:603 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 6 theme lectins 653:659 arg1 specificity 619:629 substrate specificity 609:629 substrate specificity 609:629 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 8 7 theme N278A 1162:1166 arg1 Degradation 1139:1149 Degradation 1139:1149 Degradation of SHH and N278A 1139:1166 Degradation of SHH and N278A also required OS-9, but not the related lectin XTP3-B. 24910992 11 8 theme potential 1620:1628 arg1 avenue 1630:1635 a potential avenue 1618:1635 a potential avenue by which misfolded glycoproteins may be shunted towards SEL1L and ERAD rather than being released into the secretory pathway 1618:1760 EDEM2 also interacts with calnexin and SEL1L, suggesting a potential avenue by which misfolded glycoproteins may be shunted towards SEL1L and ERAD rather than being released into the secretory pathway. 24910992 12 9 theme underlying 1883:1892 arg1 mechanism 1894:1902 an underlying mechanism 1880:1902 an underlying mechanism distinct from that of S. cerevisiae 1880:1938 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 5 10 theme ER 650:651 arg1 OS-9 683:686 OS-9 683:686 OS-9 683:686 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 10 theme ER 650:651 arg1 lectins 653:659 known mammalian ER lectins 634:659 known mammalian ER lectins 634:659 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 10 theme ER 650:651 arg1 EDEM1/2/3 672:680 EDEM1/2/3 672:680 EDEM1/2/3 672:680 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 10 theme ER 650:651 arg1 XTP-3B 692:697 XTP-3B 692:697 XTP-3B 692:697 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 11 theme identified 718:727 arg1 substrate 734:742 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 11 theme identified 718:727 arg1 protein 776:782 a soluble protein 766:782 a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A 766:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 6 12 theme N278A 875:879 arg1 ERAD 867:870 Efficient ERAD 857:870 Efficient ERAD of N278A 857:879 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 10 13 theme nonglycosylated 1478:1492 arg1 substrate 1494:1502 the first nonglycosylated substrate 1468:1502 the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD 1468:1558 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 10 13 theme nonglycosylated 1478:1492 arg1 SHH-N278A 1455:1463 SHH-N278A 1455:1463 SHH-N278A 1455:1463 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 12 14 theme distinct 1904:1911 arg1 mechanism 1894:1902 an underlying mechanism 1880:1902 an underlying mechanism distinct from that of S. cerevisiae 1880:1938 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 6 15 theme similar 942:948 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 5 16 theme ERAD 729:732 arg1 substrate 734:742 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 16 theme ERAD 729:732 arg1 protein 776:782 a soluble protein 766:782 a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A 766:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 4 17 theme similar 472:478 arg1 recognition 480:490 a similar recognition and delivery mechanism 470:513 recognition 480:490 Whether a similar recognition and delivery mechanism exists in mammalian cells is unknown. 24910992 0 18 gly non-glycosylated 61:76 arg1 hedgehog 84:91 non-glycosylated sonic hedgehog 61:91 non-glycosylated sonic hedgehog 61:91 EDEM2 and OS-9 are required for ER-associated degradation of non-glycosylated sonic hedgehog. 24910992 7 19 theme SHHs 1051:1054 arg1 ERAD 1004:1007 ERAD 1004:1007 ERAD of both glycosylated and non-glycosylated SHHs 1004:1054 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 5 20 theme substrate 609:617 arg1 specificity 619:629 substrate specificity 609:629 substrate specificity 609:629 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 2 21 theme substrate 263:271 arg1 recognition 273:283 substrate recognition 263:283 substrate recognition 263:283 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 5 22 theme mutant 843:848 arg1 N278A 850:854 its nonglycosylated mutant N278A 823:854 its nonglycosylated mutant N278A 823:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 9 23 theme glycan 1366:1371 arg1 backbone 1342:1349 the misfolded polypeptide backbone, rather than a glycan signature, 1316:1382 backbone 1342:1349 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 23 theme glycan 1366:1371 arg1 signature 1373:1381 a glycan signature 1364:1381 a glycan signature 1364:1381 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 23 theme glycan 1366:1371 arg1 signal 1413:1418 the predominant signal 1397:1418 the predominant signal for recognition for ERAD 1397:1443 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 5 24 theme sonic 744:748 arg1 SHH 760:762 SHH 760:762 SHH 760:762 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 24 theme sonic 744:748 arg1 hedgehog 750:757 sonic hedgehog 744:757 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 6 25 theme Efficient 857:865 arg1 ERAD 867:870 Efficient ERAD 857:870 Efficient ERAD of N278A 857:879 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 9 26 theme misfolded 1320:1328 arg1 backbone 1342:1349 the misfolded polypeptide backbone, rather than a glycan signature, 1316:1382 backbone 1342:1349 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 26 theme misfolded 1320:1328 arg1 signature 1373:1381 a glycan signature 1364:1381 a glycan signature 1364:1381 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 26 theme misfolded 1320:1328 arg1 signal 1413:1418 the predominant signal 1397:1418 the predominant signal for recognition for ERAD 1397:1443 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 27 gly non-glycosylated 1272:1287 arg1 variant 1293:1299 a non-glycosylated SHH variant 1270:1299 a non-glycosylated SHH variant 1270:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 5 28 theme nonglycosylated 827:841 arg1 N278A 850:854 its nonglycosylated mutant N278A 823:854 its nonglycosylated mutant N278A 823:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 12 29 theme ER 1769:1770 arg1 lectins 1772:1778 ER lectins 1769:1778 ER lectins 1769:1778 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 9 30 theme non-glycosylated 1272:1287 arg1 variant 1293:1299 a non-glycosylated SHH variant 1270:1299 a non-glycosylated SHH variant 1270:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 5 31 theme known 634:638 arg1 OS-9 683:686 OS-9 683:686 OS-9 683:686 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 31 theme known 634:638 arg1 lectins 653:659 known mammalian ER lectins 634:659 known mammalian ER lectins 634:659 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 31 theme known 634:638 arg1 EDEM1/2/3 672:680 EDEM1/2/3 672:680 EDEM1/2/3 672:680 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 31 theme known 634:638 arg1 XTP-3B 692:697 XTP-3B 692:697 XTP-3B 692:697 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 9 32 theme OS-9 1260:1263 arg1 interaction 1230:1240 Robust interaction 1223:1240 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant 1223:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 7 33 gly non-glycosylated 1034:1049 arg1 SHHs 1051:1054 both glycosylated and non-glycosylated SHHs 1012:1054 both glycosylated and non-glycosylated SHHs 1012:1054 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 1 34 theme ER-associated 169:181 arg1 ERAD 196:199 ERAD 196:199 ERAD 196:199 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 1 34 theme ER-associated 169:181 arg1 degradation 183:193 the ER-associated degradation 165:193 the ER-associated degradation (ERAD) in eukaryotes 165:214 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 6 35 theme p97 937:939 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 5 36 theme mammalian 640:648 arg1 OS-9 683:686 OS-9 683:686 OS-9 683:686 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 36 theme mammalian 640:648 arg1 lectins 653:659 known mammalian ER lectins 634:659 known mammalian ER lectins 634:659 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 36 theme mammalian 640:648 arg1 EDEM1/2/3 672:680 EDEM1/2/3 672:680 EDEM1/2/3 672:680 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 36 theme mammalian 640:648 arg1 XTP-3B 692:697 XTP-3B 692:697 XTP-3B 692:697 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 1 37 theme Misfolded 94:102 arg1 proteins 104:111 Misfolded proteins 94:111 Misfolded proteins of the endoplasmic reticulum (ER) 94:145 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 3 38 theme directional 422:432 arg1 delivery 434:441 the directional delivery 418:441 the directional delivery of the substrates 418:459 Trimming of the glycan is essential for the directional delivery of the substrates. 24910992 5 39 theme single 795:800 arg1 N-glycan 802:809 a single N-glycan 793:809 a single N-glycan 793:809 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 12 40 theme ER 1837:1838 arg1 substrates 1840:1849 misfolded ER substrates 1827:1849 misfolded ER substrates 1827:1849 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 10 41 gly nonglycosylated 1478:1492 arg1 substrate 1494:1502 the first nonglycosylated substrate 1468:1502 the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD 1468:1558 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 10 41 gly nonglycosylated 1478:1492 arg1 SHH-N278A 1455:1463 SHH-N278A 1455:1463 SHH-N278A 1455:1463 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 6 42 theme SEL1L 927:931 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 7 43 theme glycosylated 1017:1028 arg1 SHHs 1051:1054 both glycosylated and non-glycosylated SHHs 1012:1054 both glycosylated and non-glycosylated SHHs 1012:1054 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 12 44 from that 1918:1921 arg1 distinct 1904:1911 distinct 1904:1911 distinct 1904:1911 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 2 45 theme bipartite 293:301 arg1 signals 303:309 bipartite signals 293:309 bipartite signals consisting of an unfolded local structure and the adjacent glycan 293:375 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 7 46 theme non-glycosylated 1034:1049 arg1 SHHs 1051:1054 both glycosylated and non-glycosylated SHHs 1012:1054 both glycosylated and non-glycosylated SHHs 1012:1054 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 2 47 theme local 337:341 arg1 structure 343:351 an unfolded local structure 325:351 an unfolded local structure 325:351 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 9 48 theme predominant 1401:1411 arg1 backbone 1342:1349 the misfolded polypeptide backbone, rather than a glycan signature, 1316:1382 backbone 1342:1349 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 48 theme predominant 1401:1411 arg1 signature 1373:1381 a glycan signature 1364:1381 a glycan signature 1364:1381 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 48 theme predominant 1401:1411 arg1 signal 1413:1418 the predominant signal 1397:1418 the predominant signal for recognition for ERAD 1397:1443 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 8 49 theme lectin 1208:1213 arg1 XTP3-B 1215:1220 the related lectin XTP3-B 1196:1220 the related lectin XTP3-B 1196:1220 Degradation of SHH and N278A also required OS-9, but not the related lectin XTP3-B. 24910992 8 50 theme SHH 1154:1156 arg1 Degradation 1139:1149 Degradation 1139:1149 Degradation of SHH and N278A 1139:1166 Degradation of SHH and N278A also required OS-9, but not the related lectin XTP3-B. 24910992 6 51 theme HRD1 921:924 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 3 52 theme substrates 450:459 arg1 delivery 434:441 the directional delivery 418:441 the directional delivery of the substrates 418:459 Trimming of the glycan is essential for the directional delivery of the substrates. 24910992 3 53 theme glycan 394:399 arg1 Trimming 378:385 Trimming 378:385 Trimming of the glycan 378:399 Trimming of the glycan is essential for the directional delivery of the substrates. 24910992 4 54 theme mammalian 525:533 arg1 cells 535:539 mammalian cells 525:539 mammalian cells 525:539 Whether a similar recognition and delivery mechanism exists in mammalian cells is unknown. 24910992 5 55 dep substrate 734:742 arg1 SHH 760:762 SHH 760:762 SHH 760:762 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 55 dep substrate 734:742 arg1 hedgehog 750:757 sonic hedgehog 744:757 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 56 contain carrying 784:791 arg2 N278A 850:854 its nonglycosylated mutant N278A 823:854 its nonglycosylated mutant N278A 823:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 56 contain carrying 784:791 arg2 N-glycan 802:809 a single N-glycan 793:809 a single N-glycan 793:809 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 56 contain carrying 784:791 arg1 protein 776:782 a soluble protein 766:782 a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A 766:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 56 contain carrying 784:791 arg1 substrate 734:742 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 7 57 gly glycosylated 1086:1097 arg1 SHH 1099:1101 glycosylated SHH 1086:1101 glycosylated SHH 1086:1101 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 9 58 theme EDEM2 1250:1254 arg1 interaction 1230:1240 Robust interaction 1223:1240 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant 1223:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 0 59 theme ER-associated 32:44 arg1 degradation 46:56 ER-associated degradation 32:56 ER-associated degradation of non-glycosylated sonic hedgehog 32:91 EDEM2 and OS-9 are required for ER-associated degradation of non-glycosylated sonic hedgehog. 24910992 11 60 theme secretory 1744:1752 arg1 pathway 1754:1760 the secretory pathway 1740:1760 the secretory pathway 1740:1760 EDEM2 also interacts with calnexin and SEL1L, suggesting a potential avenue by which misfolded glycoproteins may be shunted towards SEL1L and ERAD rather than being released into the secretory pathway. 24910992 1 61 theme endoplasmic 120:130 arg1 reticulum 132:140 the endoplasmic reticulum 116:140 the endoplasmic reticulum (ER) 116:145 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 1 61 theme endoplasmic 120:130 arg1 ER 143:144 ER 143:144 ER 143:144 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 6 62 theme processing 899:908 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 12 63 theme misfolded 1827:1835 arg1 substrates 1840:1849 misfolded ER substrates 1827:1849 misfolded ER substrates 1827:1849 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 7 64 theme glycosylated 1086:1097 arg1 SHH 1099:1101 glycosylated SHH 1086:1101 glycosylated SHH 1086:1101 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 11 65 theme misfolded 1646:1654 arg1 glycoproteins 1656:1668 misfolded glycoproteins 1646:1668 misfolded glycoproteins 1646:1668 EDEM2 also interacts with calnexin and SEL1L, suggesting a potential avenue by which misfolded glycoproteins may be shunted towards SEL1L and ERAD rather than being released into the secretory pathway. 24910992 1 66 theme reticulum 132:140 arg1 proteins 104:111 Misfolded proteins 94:111 Misfolded proteins of the endoplasmic reticulum (ER) 94:145 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 10 67 theme first 1472:1476 arg1 substrate 1494:1502 the first nonglycosylated substrate 1468:1502 the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD 1468:1558 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 10 67 theme first 1472:1476 arg1 SHH-N278A 1455:1463 SHH-N278A 1455:1463 SHH-N278A 1455:1463 Notably, SHH-N278A is the first nonglycosylated substrate to require EDEM2 for recognition and targeting for ERAD. 24910992 2 68 theme ER-resident 235:245 arg1 lectins 247:253 ER-resident lectins 235:253 ER-resident lectins 235:253 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 12 69 theme substrates 1840:1849 arg1 delivery 1815:1822 delivery 1815:1822 delivery 1815:1822 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 12 69 theme substrates 1840:1849 arg1 recognition 1799:1809 recognition 1799:1809 recognition 1799:1809 Thus, ER lectins participate in the recognition and delivery of misfolded ER substrates differently in mammals, with an underlying mechanism distinct from that of S. cerevisiae. 24910992 9 70 theme SHH 1289:1291 arg1 variant 1293:1299 a non-glycosylated SHH variant 1270:1299 a non-glycosylated SHH variant 1270:1299 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 2 71 theme unfolded 328:335 arg1 structure 343:351 an unfolded local structure 325:351 an unfolded local structure 325:351 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 0 72 theme non-glycosylated 61:76 arg1 hedgehog 84:91 non-glycosylated sonic hedgehog 61:91 non-glycosylated sonic hedgehog 61:91 EDEM2 and OS-9 are required for ER-associated degradation of non-glycosylated sonic hedgehog. 24910992 8 73 theme related 1200:1206 arg1 XTP3-B 1215:1220 the related lectin XTP3-B 1196:1220 the related lectin XTP3-B 1196:1220 Degradation of SHH and N278A also required OS-9, but not the related lectin XTP3-B. 24910992 1 74 from degradation 183:193 arg1 eukaryotes 205:214 eukaryotes 205:214 eukaryotes 205:214 Misfolded proteins of the endoplasmic reticulum (ER) are eliminated by the ER-associated degradation (ERAD) in eukaryotes. 24910992 7 75 gly glycosylated 1017:1028 arg1 SHHs 1051:1054 both glycosylated and non-glycosylated SHHs 1012:1054 both glycosylated and non-glycosylated SHHs 1012:1054 While EDEM2 was required for ERAD of both glycosylated and non-glycosylated SHHs, EDEM3 was only necessary for glycosylated SHH and EDEM1 was dispensable for both. 24910992 5 76 dep function 596:603 arg1 the 592:594 the 592:594 the 592:594 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 9 77 theme polypeptide 1330:1340 arg1 backbone 1342:1349 the misfolded polypeptide backbone, rather than a glycan signature, 1316:1382 backbone 1342:1349 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 77 theme polypeptide 1330:1340 arg1 signature 1373:1381 a glycan signature 1364:1381 a glycan signature 1364:1381 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 9 77 theme polypeptide 1330:1340 arg1 signal 1413:1418 the predominant signal 1397:1418 the predominant signal for recognition for ERAD 1397:1443 Robust interaction of both EDEM2 and OS-9 with a non-glycosylated SHH variant indicates that the misfolded polypeptide backbone, rather than a glycan signature, functions as the predominant signal for recognition for ERAD. 24910992 5 78 theme soluble 768:774 arg1 substrate 734:742 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 the recently identified ERAD substrate sonic hedgehog (SHH) 705:763 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 5 78 theme soluble 768:774 arg1 protein 776:782 a soluble protein 766:782 a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A 766:854 In this study, we systematically study the function and substrate specificity of known mammalian ER lectins, including EDEM1/2/3, OS-9 and XTP-3B using the recently identified ERAD substrate sonic hedgehog (SHH), a soluble protein carrying a single N-glycan, as well as its nonglycosylated mutant N278A. 24910992 6 79 theme core 894:897 arg1 complex 910:916 the core processing complex 890:916 the core processing complex 890:916 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 6 80 gly glycosylated 957:968 arg1 SHH 970:972 the glycosylated SHH 953:972 the glycosylated SHH 953:972 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 24910992 2 81 theme adjacent 361:368 arg1 glycan 370:375 the adjacent glycan 357:375 the adjacent glycan 357:375 In S. cerevisiae, ER-resident lectins mediate substrate recognition through bipartite signals consisting of an unfolded local structure and the adjacent glycan. 24910992 11 82 gly glycoproteins 1656:1668 arg1 glycoproteins 1656:1668 misfolded glycoproteins 1646:1668 misfolded glycoproteins 1646:1668 EDEM2 also interacts with calnexin and SEL1L, suggesting a potential avenue by which misfolded glycoproteins may be shunted towards SEL1L and ERAD rather than being released into the secretory pathway. 24910992 6 83 theme glycosylated 957:968 arg1 SHH 970:972 the glycosylated SHH 953:972 the glycosylated SHH 953:972 Efficient ERAD of N278A requires the core processing complex of HRD1, SEL1L and p97, similar to the glycosylated SHH. 25036289 3 0 theme antigen 298:304 arg1 domains 322:328 the antigen recognition Fab domains 294:328 the antigen recognition Fab domains 294:328 The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. 25036289 5 1 theme anti-inflammatory 752:768 arg1 activity 770:777 the anti-inflammatory activity 748:777 the anti-inflammatory activity of intravenous IgG 748:796 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 8 2 theme glycan 1288:1293 arg1 sialylation 1295:1305 increased glycan sialylation 1278:1305 increased glycan sialylation 1278:1305 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 6 3 link α2,6-linked 1041:1051 arg1 acids 1060:1064 terminal α2,6-linked sialic acids 1032:1064 terminal α2,6-linked sialic acids 1032:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 8 4 gly sialylated 1226:1235 arg1 Fc 1237:1238 sialylated Fc 1226:1238 sialylated Fc 1226:1238 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 3 5 theme recognition 306:316 arg1 domains 322:328 the antigen recognition Fab domains 294:328 the antigen recognition Fab domains 294:328 The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. 25036289 7 6 theme IgG 1137:1139 arg1 Fc 1141:1142 fully disialylated IgG Fc 1118:1142 fully disialylated IgG Fc 1118:1142 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 4 7 theme inflammation 564:575 arg1 functions 551:559 its complex effector functions 530:559 its complex effector functions of inflammation, modulation, and immune suppression 530:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 4 8 theme considerable 487:498 arg1 heterogeneity 500:512 considerable heterogeneity 487:512 considerable heterogeneity 487:512 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 7 9 theme disialylated 1124:1135 arg1 Fc 1141:1142 fully disialylated IgG Fc 1118:1142 fully disialylated IgG Fc 1118:1142 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 8 10 gly asialylated 1210:1220 arg1 Fc 1222:1223 asialylated Fc 1210:1223 asialylated Fc 1210:1223 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 5 11 theme tumor 729:733 arg1 cells 735:739 tumor cells 729:739 tumor cells 729:739 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 4 12 theme Fc 454:455 arg1 domain 466:471 the Fc effector domain 450:471 the Fc effector domain 450:471 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 1 13 theme Immunoglobulin 79:92 arg1 G 94:94 Immunoglobulin G 79:94 Immunoglobulin G (IgG) 79:100 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 1 13 theme Immunoglobulin 79:92 arg1 mediator 115:122 a central mediator 105:122 a central mediator of host defense due to its ability to recognize and eliminate pathogens 105:194 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 1 13 theme Immunoglobulin 79:92 arg1 IgG 97:99 IgG 97:99 IgG 97:99 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 7 14 theme crystal 1159:1165 arg1 structure 1167:1175 its crystal structure 1155:1175 its crystal structure 1155:1175 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 2 15 theme IgGs 271:274 arg1 regions 260:266 distinct regions 251:266 distinct regions of IgGs 251:274 The recognition and effector responses are encoded on distinct regions of IgGs. 25036289 8 16 from pro-inflammatory 1412:1427 arg1 switch 1400:1405 the switch 1396:1405 the switch from pro-inflammatory to anti-inflammatory activity of the Fc 1396:1467 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 4 17 theme modulation 578:587 arg1 functions 551:559 its complex effector functions 530:559 its complex effector functions of inflammation, modulation, and immune suppression 530:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 0 18 theme proteins 69:76 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 0:77 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 6 19 theme IgG 902:904 arg1 activity 878:885 the anti-inflammatory activity 856:885 the anti-inflammatory activity of intravenous IgG 856:904 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 5 20 theme Therapeutic 614:624 arg1 antibodies 637:646 Therapeutic anti-tumor antibodies 614:646 Therapeutic anti-tumor antibodies 614:646 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 8 21 theme CH2 1366:1368 arg1 domain 1370:1375 the CH2 domain 1362:1375 the CH2 domain 1362:1375 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 22 theme asialylated 1210:1220 arg1 Fc 1222:1223 asialylated Fc 1210:1223 asialylated Fc 1210:1223 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 23 theme F241A 1245:1249 arg1 Fc 1251:1252 F241A Fc 1245:1252 F241A Fc 1245:1252 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 23 theme F241A 1245:1249 arg1 mutant 1257:1262 a mutant 1255:1262 a mutant that displays increased glycan sialylation 1255:1305 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 24 theme Fc 1237:1238 arg1 structures 1196:1205 the structures 1192:1205 the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, 1192:1306 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 6 25 theme intravenous 890:900 arg1 IgG 902:904 intravenous IgG 890:904 intravenous IgG 890:904 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 5 26 theme anti-tumor 626:635 arg1 antibodies 637:646 Therapeutic anti-tumor antibodies 614:646 Therapeutic anti-tumor antibodies 614:646 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 7 27 theme chemoenzymatic 1075:1088 arg1 glycoengineering 1090:1105 chemoenzymatic glycoengineering 1075:1105 chemoenzymatic glycoengineering 1075:1105 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 5 28 theme Fc 816:817 arg1 glycans 819:825 specific Fc glycans 807:825 specific Fc glycans 807:825 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 5 29 theme specific 807:814 arg1 glycans 819:825 specific Fc glycans 807:825 specific Fc glycans 807:825 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 4 30 theme effector 457:464 arg1 domain 466:471 the Fc effector domain 450:471 the Fc effector domain 450:471 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 6 31 theme complex 975:981 arg1 N-glycans 983:991 the Asn297-linked complex N-glycans 957:991 the Asn297-linked complex N-glycans attached to each Fc CH2 domain 957:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 0 32 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 0:77 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 4 33 theme effector 542:549 arg1 functions 551:559 its complex effector functions 530:559 its complex effector functions of inflammation, modulation, and immune suppression 530:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 8 34 theme Fc 1222:1223 arg1 structures 1196:1205 the structures 1192:1205 the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, 1192:1306 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 35 theme Fc 1251:1252 arg1 structures 1196:1205 the structures 1192:1205 the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, 1192:1306 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 0 36 theme anti-inflammatory 31:47 arg1 proteins 69:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 4 37 theme complex 534:540 arg1 functions 551:559 its complex effector functions 530:559 its complex effector functions of inflammation, modulation, and immune suppression 530:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 6 38 link Asn297-linked 961:973 arg1 N-glycans 983:991 the Asn297-linked complex N-glycans 957:991 the Asn297-linked complex N-glycans attached to each Fc CH2 domain 957:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 5 39 theme IgG 794:796 arg1 activity 770:777 the anti-inflammatory activity 748:777 the anti-inflammatory activity of intravenous IgG 748:796 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 7 40 used used 1070:1073 arg2 We 1067:1068 We 1067:1068 We 1067:1068 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 5 41 theme IgG 709:711 arg1 Fc 713:714 the IgG Fc 705:714 the IgG Fc 705:714 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 8 42 theme Fc 1466:1467 arg1 activity 1450:1457 anti-inflammatory activity 1432:1457 anti-inflammatory activity of the Fc 1432:1467 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 43 theme increased 1322:1330 arg1 flexibility 1347:1357 increased conformational flexibility 1322:1357 increased conformational flexibility of the CH2 domain 1322:1375 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 6 44 theme sialic 1053:1058 arg1 acids 1060:1064 terminal α2,6-linked sialic acids 1032:1064 terminal α2,6-linked sialic acids 1032:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 0 45 theme G 64:64 arg1 proteins 69:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 8 46 theme increased 1278:1286 arg1 sialylation 1295:1305 increased glycan sialylation 1278:1305 increased glycan sialylation 1278:1305 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 5 47 theme Fc 713:714 arg1 properties 691:700 the pro-inflammatory properties 670:700 the pro-inflammatory properties of the IgG Fc 670:714 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 6 48 theme anti-inflammatory 860:876 arg1 activity 878:885 the anti-inflammatory activity 856:885 the anti-inflammatory activity of intravenous IgG 856:904 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 6 49 theme α2,6-linked 1041:1051 arg1 acids 1060:1064 terminal α2,6-linked sialic acids 1032:1064 terminal α2,6-linked sialic acids 1032:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 0 50 theme immunoglobulin 49:62 arg1 proteins 69:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 5 51 theme intravenous 782:792 arg1 IgG 794:796 intravenous IgG 782:796 intravenous IgG 782:796 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 2 52 theme effector 217:224 arg1 responses 226:234 The recognition and effector responses 197:234 The recognition and effector responses 197:234 The recognition and effector responses are encoded on distinct regions of IgGs. 25036289 6 53 theme terminal 1032:1039 arg1 acids 1060:1064 terminal α2,6-linked sialic acids 1032:1064 terminal α2,6-linked sialic acids 1032:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 3 54 theme high 370:373 arg1 specificity 375:385 high specificity 370:385 high specificity 370:385 The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. 25036289 6 55 theme IgGs 943:946 arg1 population 929:938 a small population 921:938 a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids 921:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 6 56 theme Asn297-linked 961:973 arg1 N-glycans 983:991 the Asn297-linked complex N-glycans 957:991 the Asn297-linked complex N-glycans attached to each Fc CH2 domain 957:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 8 57 theme domain 1370:1375 arg1 flexibility 1347:1357 increased conformational flexibility 1322:1357 increased conformational flexibility of the CH2 domain 1322:1375 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 1 58 theme central 107:113 arg1 G 94:94 Immunoglobulin G 79:94 Immunoglobulin G (IgG) 79:100 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 1 58 theme central 107:113 arg1 mediator 115:122 a central mediator 105:122 a central mediator of host defense due to its ability to recognize and eliminate pathogens 105:194 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 0 59 theme Fc 66:67 arg1 proteins 69:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 anti-inflammatory immunoglobulin G Fc proteins 31:76 Structural characterization of anti-inflammatory immunoglobulin G Fc proteins. 25036289 5 60 theme pro-inflammatory 674:689 arg1 properties 691:700 the pro-inflammatory properties 670:700 the pro-inflammatory properties of the IgG Fc 670:714 Therapeutic anti-tumor antibodies, for example, require the pro-inflammatory properties of the IgG Fc to eliminate tumor cells, while the anti-inflammatory activity of intravenous IgG requires specific Fc glycans for activity. 25036289 2 61 theme recognition 201:211 arg1 responses 226:234 The recognition and effector responses 197:234 The recognition and effector responses 197:234 The recognition and effector responses are encoded on distinct regions of IgGs. 25036289 8 62 theme sialylated 1226:1235 arg1 Fc 1237:1238 sialylated Fc 1226:1238 sialylated Fc 1226:1238 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 8 63 theme anti-inflammatory 1432:1448 arg1 activity 1450:1457 anti-inflammatory activity 1432:1457 anti-inflammatory activity of the Fc 1432:1467 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 2 64 theme distinct 251:258 arg1 regions 260:266 distinct regions 251:266 distinct regions of IgGs 251:274 The recognition and effector responses are encoded on distinct regions of IgGs. 25036289 4 65 theme immune 594:599 arg1 suppression 601:611 immune suppression 594:611 immune suppression 594:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 6 66 attach attached 993:1000 arg2 N-glycans 983:991 the Asn297-linked complex N-glycans 957:991 the Asn297-linked complex N-glycans attached to each Fc CH2 domain 957:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 6 66 attach attached 993:1000 arg1 domain 1017:1022 each Fc CH2 domain 1005:1022 each Fc CH2 domain 1005:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 4 67 theme Recent 415:420 arg1 studies 422:428 Recent studies 415:428 Recent studies 415:428 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 25036289 8 68 theme structures 1196:1205 arg1 Comparison 1178:1187 Comparison 1178:1187 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, 1178:1306 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 1 69 theme host 127:130 arg1 defense 132:138 host defense 127:138 host defense 127:138 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 8 70 theme conformational 1332:1345 arg1 flexibility 1347:1357 increased conformational flexibility 1322:1357 increased conformational flexibility of the CH2 domain 1322:1375 Comparison of the structures of asialylated Fc, sialylated Fc, and F241A Fc, a mutant that displays increased glycan sialylation, suggests that increased conformational flexibility of the CH2 domain is associated with the switch from pro-inflammatory to anti-inflammatory activity of the Fc. 25036289 3 71 theme Fab 318:320 arg1 domains 322:328 the antigen recognition Fab domains 294:328 the antigen recognition Fab domains 294:328 The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. 25036289 7 72 gly disialylated 1124:1135 arg1 Fc 1141:1142 fully disialylated IgG Fc 1118:1142 fully disialylated IgG Fc 1118:1142 We used chemoenzymatic glycoengineering to prepare fully disialylated IgG Fc and solved its crystal structure. 25036289 1 73 theme defense 132:138 arg1 G 94:94 Immunoglobulin G 79:94 Immunoglobulin G (IgG) 79:100 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 1 73 theme defense 132:138 arg1 mediator 115:122 a central mediator 105:122 a central mediator of host defense due to its ability to recognize and eliminate pathogens 105:194 Immunoglobulin G (IgG) is a central mediator of host defense due to its ability to recognize and eliminate pathogens. 25036289 6 74 theme CH2 1013:1015 arg1 domain 1017:1022 each Fc CH2 domain 1005:1022 each Fc CH2 domain 1005:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 3 75 theme domains 322:328 arg1 diversity 281:289 The diversity 277:289 The diversity of the antigen recognition Fab domains 277:328 The diversity of the antigen recognition Fab domains accounts for IgG's ability to bind with high specificity to essentially any antigen. 25036289 6 76 theme small 923:927 arg1 population 929:938 a small population 921:938 a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids 921:1064 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 6 77 theme Fc 1010:1011 arg1 domain 1017:1022 each Fc CH2 domain 1005:1022 each Fc CH2 domain 1005:1022 In particular, the anti-inflammatory activity of intravenous IgG is ascribed to a small population of IgGs in which the Asn297-linked complex N-glycans attached to each Fc CH2 domain include terminal α2,6-linked sialic acids. 25036289 4 78 theme suppression 601:611 arg1 functions 551:559 its complex effector functions 530:559 its complex effector functions of inflammation, modulation, and immune suppression 530:611 Recent studies have indicated that the Fc effector domain also displays considerable heterogeneity, accounting for its complex effector functions of inflammation, modulation, and immune suppression. 28075586 0 0 theme Automated 121:129 arg1 Assembly 138:145 Automated Glycan Assembly 121:145 Automated Glycan Assembly 121:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 28075586 3 1 theme glycan 370:375 arg1 assembly 377:384 the automated glycan assembly 356:384 the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 356:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 1 2 from applications 223:234 arg1 products 244:251 food products 239:251 food products 239:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 4 3 theme oligosaccharides 673:688 arg1 pattern 658:664 arabinose substitution pattern 635:664 arabinose substitution pattern of the oligosaccharides 635:688 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 4 3 theme oligosaccharides 673:688 arg1 requirements 602:613 different requirements 592:613 different requirements for the lengths 592:629 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 4 4 theme products 574:581 arg1 Analysis 548:555 Analysis 548:555 Analysis of the digestion products 548:581 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 1 5 theme many 207:210 arg1 applications 223:234 many industrial applications 207:234 many industrial applications in food products 207:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 4 6 theme different 592:600 arg1 requirements 602:613 different requirements 592:613 different requirements for the lengths 592:629 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 3 7 theme arabinogalactan 421:435 arg1 chains 442:447 the arabinogalactan side chains 417:447 the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 417:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 1 8 with polysaccharide 187:200 arg1 applications 223:234 many industrial applications 207:234 many industrial applications in food products 207:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 3 9 theme related 406:412 arg1 oligosaccharides 389:404 oligosaccharides 389:404 oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 389:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 2 10 theme glycosyl 325:332 arg1 hydrolases 334:343 glycosyl hydrolases 325:343 glycosyl hydrolases 325:343 The structural elucidation of pectin is aided by digestion assays with glycosyl hydrolases. 28075586 1 11 theme industrial 212:221 arg1 applications 223:234 many industrial applications 207:234 many industrial applications in food products 207:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 3 12 theme novel 462:466 arg1 tools 480:484 novel biochemical tools 462:484 novel biochemical tools to determine the substrate specificities of endogalactanases 462:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 4 13 theme digestion 564:572 arg1 products 574:581 the digestion products 560:581 the digestion products 560:581 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 3 14 theme substrate 503:511 arg1 specificities 513:525 the substrate specificities 499:525 the substrate specificities of endogalactanases 499:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 0 15 theme Substrate 12:20 arg1 Specificities 22:34 Substrate Specificities 12:34 Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly 12:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 28075586 3 16 theme pectin 452:457 arg1 chains 442:447 the arabinogalactan side chains 417:447 the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 417:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 4 17 theme substitution 645:656 arg1 pattern 658:664 arabinose substitution pattern 635:664 arabinose substitution pattern of the oligosaccharides 635:688 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 3 18 dep tools 480:484 arg1 determine 489:497 determine 489:497 to determine the substrate specificities of endogalactanases 486:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 1 19 theme food 239:242 arg1 products 244:251 food products 239:251 food products 239:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 3 20 theme side 437:440 arg1 chains 442:447 the arabinogalactan side chains 417:447 the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 417:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 4 21 theme arabinose 635:643 arg1 pattern 658:664 arabinose substitution pattern 635:664 arabinose substitution pattern of the oligosaccharides 635:688 Analysis of the digestion products revealed different requirements for the lengths and arabinose substitution pattern of the oligosaccharides to be recognized and hydrolyzed by the galactanases. 28075586 3 22 theme endogalactanases 530:545 arg1 specificities 513:525 the substrate specificities 499:525 the substrate specificities of endogalactanases 499:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 3 23 theme biochemical 468:478 arg1 tools 480:484 novel biochemical tools 462:484 novel biochemical tools to determine the substrate specificities of endogalactanases 462:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 0 24 theme β1,4-Endogalactanases 39:59 arg1 Specificities 22:34 Substrate Specificities 12:34 Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly 12:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 28075586 2 25 theme pectin 284:289 arg1 elucidation 269:279 The structural elucidation 254:279 The structural elucidation of pectin 254:289 The structural elucidation of pectin is aided by digestion assays with glycosyl hydrolases. 28075586 0 26 theme Arabinogalactan 73:87 arg1 Oligosaccharides 89:104 Plant Arabinogalactan Oligosaccharides 67:104 Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly 67:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 28075586 0 27 theme Plant 67:71 arg1 Oligosaccharides 89:104 Plant Arabinogalactan Oligosaccharides 67:104 Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly 67:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 28075586 2 28 with assays 313:318 arg1 hydrolases 334:343 glycosyl hydrolases 325:343 glycosyl hydrolases 325:343 The structural elucidation of pectin is aided by digestion assays with glycosyl hydrolases. 28075586 3 29 theme automated 360:368 arg1 assembly 377:384 the automated glycan assembly 356:384 the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 356:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 2 30 theme structural 258:267 arg1 elucidation 269:279 The structural elucidation 254:279 The structural elucidation of pectin 254:289 The structural elucidation of pectin is aided by digestion assays with glycosyl hydrolases. 28075586 2 31 theme digestion 303:311 arg1 assays 313:318 digestion assays 303:318 digestion assays with glycosyl hydrolases 303:343 The structural elucidation of pectin is aided by digestion assays with glycosyl hydrolases. 28075586 1 32 theme complex 173:179 arg1 polysaccharide 187:200 a structurally complex plant polysaccharide 158:200 a structurally complex plant polysaccharide with many industrial applications in food products 158:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 1 32 theme complex 173:179 arg1 Pectin 148:153 Pectin 148:153 Pectin 148:153 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 1 33 theme plant 181:185 arg1 polysaccharide 187:200 a structurally complex plant polysaccharide 158:200 a structurally complex plant polysaccharide with many industrial applications in food products 158:251 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 1 33 theme plant 181:185 arg1 Pectin 148:153 Pectin 148:153 Pectin 148:153 Pectin is a structurally complex plant polysaccharide with many industrial applications in food products. 28075586 3 34 theme oligosaccharides 389:404 arg1 assembly 377:384 the automated glycan assembly 356:384 the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases 356:545 We report the automated glycan assembly of oligosaccharides related to the arabinogalactan side chains of pectin as novel biochemical tools to determine the substrate specificities of endogalactanases. 28075586 0 35 theme Glycan 131:136 arg1 Assembly 138:145 Automated Glycan Assembly 121:145 Automated Glycan Assembly 121:145 Determining Substrate Specificities of β1,4-Endogalactanases Using Plant Arabinogalactan Oligosaccharides Synthesized by Automated Glycan Assembly. 25196296 1 0 theme therapeutic 280:290 arg1 applications 292:303 several therapeutic applications 272:303 several therapeutic applications 272:303 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 3 1 theme therapeutic 428:438 arg1 antibodies 440:449 three therapeutic antibodies 422:449 three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana 422:518 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 3 1 theme therapeutic 428:438 arg1 Adalimumab 536:545 Adalimumab 536:545 Adalimumab 536:545 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 3 1 theme therapeutic 428:438 arg1 Infliximab 521:530 Infliximab 521:530 Infliximab 521:530 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 3 2 from cytokines 485:493 arg1 benthamiana 508:518 Nicotiana benthamiana 498:518 Nicotiana benthamiana 498:518 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 5 3 theme expression 739:748 arg1 levels 729:734 comparable levels 718:734 comparable levels of expression 718:748 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 11 4 theme recombinant 1644:1654 arg1 protein 1656:1662 a plant-produced recombinant protein 1627:1662 a plant-produced recombinant protein 1627:1662 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 10 5 theme core 1454:1457 arg1 α1,3-fucose 1459:1469 core α1,3-fucose 1454:1469 core α1,3-fucose 1454:1469 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 10 6 theme IgA-based 1405:1413 arg1 antibodies 1415:1424 the IgA-based antibodies 1401:1424 the IgA-based antibodies 1401:1424 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 3 7 theme antibodies 440:449 arg1 versions 410:417 IgA1κ and IgG1κ versions 394:417 IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit 394:634 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 11 8 theme N-glycan 1522:1529 arg1 type 1514:1517 This type 1509:1517 This type of N-glycan 1509:1529 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 11 8 theme N-glycan 1522:1529 arg1 fraction 1615:1622 the main fraction 1606:1622 the main fraction of a plant-produced recombinant protein 1606:1662 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 6 9 theme IgG 957:959 arg1 counterparts 961:972 their IgG counterparts 951:972 their IgG counterparts 951:972 However, IgA1κ-based Adalimumab and Ustekinumab were poorly secreted compared to their IgG counterparts. 25196296 8 10 theme N-glycan 1156:1163 arg1 structures 1165:1174 oligomannose-type N-glycan structures 1138:1174 oligomannose-type N-glycan structures 1138:1174 This corresponded with the observation that both IgA1κ- and IgG1κ-based Infliximab were enriched in oligomannose-type N-glycan structures. 25196296 2 11 from scarce 374:379 arg1 contrast 318:325 contrast 318:325 contrast to IgG 318:332 However, in contrast to IgG, reports on plant production of IgA are scarce. 25196296 5 12 contain have 820:823 arg1 antibodies 703:712 All six antibodies 695:712 All six antibodies 695:712 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 5 12 contain have 820:823 arg2 activity 838:845 neutralizing activity 825:845 neutralizing activity 825:845 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 9 13 with biantennary 1285:1295 arg1 β1,2-xylose 1332:1342 β1,2-xylose 1332:1342 β1,2-xylose 1332:1342 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 13 with biantennary 1285:1295 arg1 α1,3-fucose 1353:1363 core α1,3-fucose 1348:1363 core α1,3-fucose 1348:1363 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 13 with biantennary 1285:1295 arg1 N-acetylglucosamine 1311:1329 terminal N-acetylglucosamine 1302:1329 terminal N-acetylglucosamine 1302:1329 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 8 14 theme oligomannose-type 1138:1154 arg1 structures 1165:1174 oligomannose-type N-glycan structures 1138:1174 oligomannose-type N-glycan structures 1138:1174 This corresponded with the observation that both IgA1κ- and IgG1κ-based Infliximab were enriched in oligomannose-type N-glycan structures. 25196296 5 15 contain had 714:716 arg1 antibodies 703:712 All six antibodies 695:712 All six antibodies 695:712 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 5 15 contain had 714:716 arg2 levels 729:734 comparable levels 718:734 comparable levels of expression 718:748 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 11 16 theme marginal 1549:1556 arg1 percentages 1558:1568 marginal percentages 1549:1568 marginal percentages in plants 1549:1578 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 3 17 theme Nicotiana 498:506 arg1 benthamiana 508:518 Nicotiana benthamiana 498:518 Nicotiana benthamiana 498:518 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 10 18 from contrast 1369:1376 arg1 N-glycan 1389:1396 the major N-glycan 1379:1396 the major N-glycan on the IgA-based antibodies 1379:1424 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 10 18 from contrast 1369:1376 arg1 xylosylated 1430:1440 xylosylated 1430:1440 xylosylated 1430:1440 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 3 19 theme pro-inflammatory 468:483 arg1 cytokines 485:493 pro-inflammatory cytokines 468:493 pro-inflammatory cytokines in Nicotiana benthamiana 468:518 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 0 20 theme different 74:82 arg1 N-glycans 84:92 different N-glycans 74:92 different N-glycans 74:92 Monomeric IgA can be produced in planta as efficient as IgG, yet receives different N-glycans. 25196296 10 21 theme terminal 1479:1486 arg1 N-acetylglucosamine 1488:1506 one terminal N-acetylglucosamine 1475:1506 one terminal N-acetylglucosamine 1475:1506 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 11 22 theme plant-produced 1629:1642 arg1 protein 1656:1662 a plant-produced recombinant protein 1627:1662 a plant-produced recombinant protein 1627:1662 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 12 23 theme receives 1778:1785 arg1 type 1749:1752 the type 1745:1752 the type of N-glycan an antibody receives 1745:1785 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 7 24 theme isotype 1020:1026 arg1 backbone 1028:1035 isotype backbone 1020:1035 isotype backbone 1020:1035 Infliximab was poorly secreted regardless of isotype backbone. 25196296 0 25 theme Monomeric 0:8 arg1 IgA 10:12 Monomeric IgA 0:12 Monomeric IgA 0:12 Monomeric IgA can be produced in planta as efficient as IgG, yet receives different N-glycans. 25196296 9 26 theme core 1348:1351 arg1 α1,3-fucose 1353:1363 core α1,3-fucose 1348:1363 core α1,3-fucose 1348:1363 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 12 27 theme N-glycan 1757:1764 arg1 receives 1778:1785 N-glycan an antibody receives 1757:1785 N-glycan an antibody receives 1757:1785 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 3 28 dep antibodies 440:449 arg1 antibodies 440:449 three therapeutic antibodies 422:449 three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana 422:518 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 3 28 dep antibodies 440:449 arg1 Adalimumab 536:545 Adalimumab 536:545 Adalimumab 536:545 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 3 28 dep antibodies 440:449 arg1 Infliximab 521:530 Infliximab 521:530 Infliximab 521:530 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 5 29 theme neutralizing 825:836 arg1 activity 838:845 neutralizing activity 825:845 neutralizing activity 825:845 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 5 30 theme total 772:776 arg1 content 794:800 total soluble protein content 772:800 total soluble protein content 772:800 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 9 31 theme major 1225:1229 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 31 theme major 1225:1229 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 12 32 theme antibody 1769:1776 arg1 receives 1778:1785 N-glycan an antibody receives 1757:1785 N-glycan an antibody receives 1757:1785 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 3 33 theme IgA1κ 394:398 arg1 versions 410:417 IgA1κ and IgG1κ versions 394:417 IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit 394:634 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 2 34 theme IgA 366:368 arg1 production 352:361 plant production 346:361 plant production of IgA 346:368 However, in contrast to IgG, reports on plant production of IgA are scarce. 25196296 9 35 theme N-glycan 1231:1238 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 35 theme N-glycan 1231:1238 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 5 36 theme cell-based 852:861 arg1 assay 863:867 a cell-based assay 850:867 a cell-based assay 850:867 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 5 37 theme content 794:800 arg1 content 794:800 total soluble protein content 772:800 total soluble protein content 772:800 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 5 37 theme content 794:800 arg1 % 767:767 3.5 and 9% 758:767 3.5 and 9% of total soluble protein content 758:800 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 12 38 contain have 1716:1719 arg2 influence 1732:1740 a profound influence 1721:1740 a profound influence 1721:1740 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 12 38 contain have 1716:1719 arg1 isotype 1704:1710 the antibody isotype 1691:1710 the antibody isotype 1691:1710 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 9 39 theme biantennary 1285:1295 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 39 theme biantennary 1285:1295 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 2 40 theme plant 346:350 arg1 production 352:361 plant production 346:361 plant production of IgA 346:368 However, in contrast to IgG, reports on plant production of IgA are scarce. 25196296 1 41 theme promising 241:249 arg1 isotype 260:266 a promising antibody isotype 239:266 a promising antibody isotype for several therapeutic applications 239:303 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 11 42 theme protein 1656:1662 arg1 type 1514:1517 This type 1509:1517 This type of N-glycan 1509:1529 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 11 42 theme protein 1656:1662 arg1 fraction 1615:1622 the main fraction 1606:1622 the main fraction of a plant-produced recombinant protein 1606:1662 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 1 43 theme antibody 251:258 arg1 isotype 260:266 a promising antibody isotype 239:266 a promising antibody isotype for several therapeutic applications 239:303 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 11 44 theme main 1610:1613 arg1 type 1514:1517 This type 1509:1517 This type of N-glycan 1509:1529 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 11 44 theme main 1610:1613 arg1 fraction 1615:1622 the main fraction 1606:1622 the main fraction of a plant-produced recombinant protein 1606:1662 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 9 45 theme terminal 1302:1309 arg1 N-acetylglucosamine 1311:1329 terminal N-acetylglucosamine 1302:1329 terminal N-acetylglucosamine 1302:1329 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 3 46 theme IgG1κ 404:408 arg1 versions 410:417 IgA1κ and IgG1κ versions 394:417 IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit 394:634 We produced IgA1κ and IgG1κ versions of three therapeutic antibodies directed against pro-inflammatory cytokines in Nicotiana benthamiana: Infliximab and Adalimumab, directed against TNF-α, and Ustekinumab, directed against the interleukin-12p40 subunit. 25196296 9 47 theme typical 1253:1259 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 47 theme typical 1253:1259 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 12 48 theme antibody 1695:1702 arg1 isotype 1704:1710 the antibody isotype 1691:1710 the antibody isotype 1691:1710 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 2 49 from reports 335:341 arg1 production 352:361 plant production 346:361 plant production of IgA 346:368 However, in contrast to IgG, reports on plant production of IgA are scarce. 25196296 10 50 from N-glycan 1389:1396 arg1 antibodies 1415:1424 the IgA-based antibodies 1401:1424 the IgA-based antibodies 1401:1424 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 12 51 theme profound 1723:1730 arg1 influence 1732:1740 a profound influence 1721:1740 a profound influence 1721:1740 Our data demonstrate that the antibody isotype may have a profound influence on the type of N-glycan an antibody receives. 25196296 6 52 theme IgA1κ-based 879:889 arg1 Adalimumab 891:900 IgA1κ-based Adalimumab 879:900 IgA1κ-based Adalimumab 879:900 However, IgA1κ-based Adalimumab and Ustekinumab were poorly secreted compared to their IgG counterparts. 25196296 8 53 theme IgG1κ-based 1098:1108 arg1 Infliximab 1110:1119 IgG1κ-based Infliximab 1098:1119 IgG1κ-based Infliximab 1098:1119 This corresponded with the observation that both IgA1κ- and IgG1κ-based Infliximab were enriched in oligomannose-type N-glycan structures. 25196296 9 54 theme plant 1261:1265 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 54 theme plant 1261:1265 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 55 theme IgG1κ-based 1181:1191 arg1 Ustekinumab 1193:1203 IgG1κ-based Ustekinumab 1181:1203 IgG1κ-based Ustekinumab 1181:1203 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 1 56 theme unique 99:104 arg1 ability 135:141 the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE 131:228 the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE 131:228 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 1 56 theme unique 99:104 arg1 features 106:113 The unique features 95:113 The unique features 95:113 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 9 57 theme complex 1267:1273 arg1 type 1240:1243 the major N-glycan type 1221:1243 the major N-glycan type 1221:1243 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 9 57 theme complex 1267:1273 arg1 N-glycan 1275:1282 the typical plant complex N-glycan 1249:1282 the typical plant complex N-glycan 1249:1282 For IgG1κ-based Ustekinumab and Adalimumab, the major N-glycan type was the typical plant complex N-glycan, biantennary with terminal N-acetylglucosamine, β1,2-xylose and core α1,3-fucose. 25196296 10 58 theme major 1383:1387 arg1 N-glycan 1389:1396 the major N-glycan 1379:1396 the major N-glycan on the IgA-based antibodies 1379:1424 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 10 58 theme major 1383:1387 arg1 xylosylated 1430:1440 xylosylated 1430:1440 xylosylated 1430:1440 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 11 59 from percentages 1558:1568 arg1 plants 1573:1578 plants 1573:1578 plants 1573:1578 This type of N-glycan occurs usually in marginal percentages in plants and was never shown to be the main fraction of a plant-produced recombinant protein. 25196296 1 60 theme inflammatory 183:194 arg1 responses 196:204 the inflammatory responses 179:204 the inflammatory responses mediated by IgG and IgE 179:228 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 5 61 theme comparable 718:727 arg1 levels 729:734 comparable levels 718:734 comparable levels of expression 718:748 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 10 62 from xylosylated 1430:1440 arg1 contrast 1369:1376 contrast 1369:1376 contrast 1369:1376 In contrast, the major N-glycan on the IgA-based antibodies was xylosylated, but lacked core α1,3-fucose and one terminal N-acetylglucosamine. 25196296 5 63 theme protein 786:792 arg1 content 794:800 total soluble protein content 772:800 total soluble protein content 772:800 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 4 64 theme antibody 650:657 arg1 yield 659:663 antibody yield 650:663 antibody yield 650:663 We evaluated antibody yield, quality and N-glycosylation. 25196296 1 65 theme several 272:278 arg1 applications 292:303 several therapeutic applications 272:303 several therapeutic applications 272:303 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 5 66 theme soluble 778:784 arg1 content 794:800 total soluble protein content 772:800 total soluble protein content 772:800 All six antibodies had comparable levels of expression between 3.5 and 9% of total soluble protein content and were shown to have neutralizing activity in a cell-based assay. 25196296 1 67 theme IgA 118:120 arg1 ability 135:141 the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE 131:228 the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE 131:228 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 25196296 1 67 theme IgA 118:120 arg1 features 106:113 The unique features 95:113 The unique features 95:113 The unique features of IgA, such as the ability to recruit neutrophils and suppress the inflammatory responses mediated by IgG and IgE, make it a promising antibody isotype for several therapeutic applications. 27320861 10 0 dep O. 1504:1505 arg1 sativa 1507:1512 O. sativa 1504:1512 O. sativa 1504:1512 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 7 1 from structures 1153:1162 arg1 seedlings 1177:1185 O. sativa seedlings 1167:1185 O. sativa seedlings 1167:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 5 2 from structures 869:878 arg1 root 897:900 root 897:900 root 897:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 5 2 from structures 869:878 arg1 shoot 887:891 shoot 887:891 shoot 887:891 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 10 3 theme O. 1504:1505 arg1 root 1496:1499 root 1496:1499 root 1496:1499 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 3 theme O. 1504:1505 arg1 shoot 1486:1490 shoot 1486:1490 shoot 1486:1490 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 7 4 theme O. 1223:1224 arg1 seedlings 1233:1241 O. sativa seedlings 1223:1241 O. sativa seedlings grown in the dark 1223:1259 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 0 5 from transition 9:18 arg1 sativa 62:67 Oryza sativa 56:67 Oryza sativa 56:67 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 7 6 theme light 1135:1139 arg1 effects 1124:1130 the effects 1120:1130 the effects of light on N-glycan structures in O. sativa seedlings 1120:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 5 7 from root 897:900 arg1 analyses 853:860 comparative analyses 841:860 comparative analyses of the structures in the shoot and root 841:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 7 8 theme N-glycan 1200:1207 arg1 structures 1209:1218 N-glycan structures 1200:1218 N-glycan structures 1200:1218 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 11 9 theme synthetic 1619:1627 arg1 system 1629:1634 the N-glycan synthetic system 1606:1634 the N-glycan synthetic system of the O. sativa shoot 1606:1657 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 4 10 theme main 377:380 arg1 type 382:385 the main type 373:385 the main type of N-glycan transition 373:408 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 10 11 theme synthetic 1565:1573 arg1 systems 1575:1581 distinct N-glycan synthetic systems 1547:1581 distinct N-glycan synthetic systems 1547:1581 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 12 theme N-glycan 1556:1563 arg1 systems 1575:1581 distinct N-glycan synthetic systems 1547:1581 distinct N-glycan synthetic systems 1547:1581 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 11 13 theme sativa 1646:1651 arg1 shoot 1653:1657 the O. sativa shoot 1639:1657 the O. sativa shoot 1639:1657 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 4 14 theme N-glycan 390:397 arg1 transition 399:408 N-glycan transition 390:408 N-glycan transition 390:408 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 10 15 contain have 1514:1517 arg2 systems 1575:1581 distinct N-glycan synthetic systems 1547:1581 distinct N-glycan synthetic systems 1547:1581 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 15 contain have 1514:1517 arg1 shoot 1486:1490 shoot 1486:1490 shoot 1486:1490 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 15 contain have 1514:1517 arg2 glycoproteins 1529:1541 different glycoproteins 1519:1541 different glycoproteins 1519:1541 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 15 contain have 1514:1517 arg1 root 1496:1499 root 1496:1499 root 1496:1499 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 10 16 theme distinct 1547:1554 arg1 systems 1575:1581 distinct N-glycan synthetic systems 1547:1581 distinct N-glycan synthetic systems 1547:1581 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 11 17 theme shoot 1653:1657 arg1 system 1629:1634 the N-glycan synthetic system 1606:1634 the N-glycan synthetic system of the O. sativa shoot 1606:1657 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 4 18 from germination 471:481 arg1 sativa 492:497 Oryza sativa 486:497 Oryza sativa 486:497 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 8 19 from expression 1287:1296 arg1 root 1305:1308 the root 1301:1308 the root 1301:1308 Understandably, N-glycan expression in the root was almost unaffected by light. 27320861 7 20 theme N-glycan 1144:1151 arg1 structures 1153:1162 N-glycan structures 1144:1162 N-glycan structures in O. sativa seedlings 1144:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 11 21 theme photosynthesis-related 1714:1735 arg1 proteins 1737:1744 many photosynthesis-related proteins 1709:1744 many photosynthesis-related proteins 1709:1744 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 5 22 theme O. 778:779 arg1 seedlings 788:796 O. sativa seedlings 778:796 O. sativa seedlings grown under light conditions 778:825 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 5 23 theme light 810:814 arg1 conditions 816:825 light conditions 810:825 light conditions 810:825 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 5 24 theme structures 869:878 arg1 analyses 853:860 comparative analyses 841:860 comparative analyses of the structures in the shoot and root 841:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 11 25 theme O. 1643:1644 arg1 shoot 1653:1657 the O. sativa shoot 1639:1657 the O. sativa shoot 1639:1657 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 4 26 gly glycoproteins 560:572 arg1 glycoproteins 560:572 glycoproteins 560:572 glycoproteins 560:572 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 6 27 theme light 1078:1082 arg1 conditions 1084:1093 light conditions 1078:1093 light conditions 1078:1093 The analyses show that fundamental N-glycan structures are common to the shoot and root, whereas paucimannosidic-type N-glycans dramatically decreased in the root grown under light conditions. 27320861 0 28 theme N-glycan 0:7 arg1 transition 9:18 N-glycan transition 0:18 N-glycan transition of the early developmental stage in Oryza sativa 0:67 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 9 29 theme marked 1361:1366 arg1 difference 1368:1377 a marked difference 1359:1377 a marked difference in phenotype 1359:1390 However, despite a marked difference in phenotype, N-glycan expression in the shoot was also unaffected by light. 27320861 11 30 theme N-glycan 1610:1617 arg1 system 1629:1634 the N-glycan synthetic system 1606:1634 the N-glycan synthetic system of the O. sativa shoot 1606:1657 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 9 31 theme N-glycan 1393:1400 arg1 expression 1402:1411 N-glycan expression 1393:1411 N-glycan expression in the shoot 1393:1424 However, despite a marked difference in phenotype, N-glycan expression in the shoot was also unaffected by light. 27320861 7 32 theme O. 1167:1168 arg1 seedlings 1177:1185 O. sativa seedlings 1167:1185 O. sativa seedlings 1167:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 4 33 theme N-glycan 531:538 arg1 transitions 516:526 transitions 516:526 transitions 516:526 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 4 33 theme N-glycan 531:538 arg1 those 551:555 those 551:555 those 551:555 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 5 34 from shoot 887:891 arg1 analyses 853:860 comparative analyses 841:860 comparative analyses of the structures in the shoot and root 841:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 5 35 dep shoot 887:891 arg1 the 883:885 the 883:885 the 883:885 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 4 36 theme plant 628:632 arg1 growth 634:639 plant growth 628:639 plant growth 628:639 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 7 37 theme sativa 1170:1175 arg1 seedlings 1177:1185 O. sativa seedlings 1167:1185 O. sativa seedlings 1167:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 5 38 theme plant 711:715 arg1 growth 717:722 plant growth 711:722 plant growth 711:722 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 10 39 theme different 1519:1527 arg1 glycoproteins 1529:1541 different glycoproteins 1519:1541 different glycoproteins 1519:1541 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 5 40 theme comparative 841:851 arg1 analyses 853:860 comparative analyses 841:860 comparative analyses of the structures in the shoot and root 841:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 0 41 theme developmental 33:45 arg1 stage 47:51 the early developmental stage 23:51 the early developmental stage 23:51 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 2 42 attach linked 156:161 arg2 N-glycans 146:154 N-glycans 146:154 N-glycans linked to proteins 146:173 In animals, N-glycans linked to proteins function in cell-cell recognition, sorting, transport, and other biological phenomena. 27320861 2 42 attach linked 156:161 arg1 proteins 166:173 proteins 166:173 proteins 166:173 In animals, N-glycans linked to proteins function in cell-cell recognition, sorting, transport, and other biological phenomena. 27320861 9 43 from expression 1402:1411 arg1 shoot 1420:1424 the shoot 1416:1424 the shoot 1416:1424 However, despite a marked difference in phenotype, N-glycan expression in the shoot was also unaffected by light. 27320861 11 44 mod modified 1754:1761 arg1 proteins 1737:1744 many photosynthesis-related proteins 1709:1744 many photosynthesis-related proteins 1709:1744 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 11 44 mod modified 1754:1761 arg3 N-glycans 1766:1774 N-glycans 1766:1774 N-glycans 1766:1774 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 8 45 theme N-glycan 1278:1285 arg1 expression 1287:1296 N-glycan expression 1278:1296 N-glycan expression in the root 1278:1308 Understandably, N-glycan expression in the root was almost unaffected by light. 27320861 0 46 theme early 27:31 arg1 stage 47:51 the early developmental stage 23:51 the early developmental stage 23:51 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 5 47 theme sativa 781:786 arg1 seedlings 788:796 O. sativa seedlings 778:796 O. sativa seedlings grown under light conditions 778:825 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 4 48 theme previous 342:349 arg1 study 351:355 a previous study 340:355 a previous study 340:355 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 7 49 from effects 1124:1130 arg1 structures 1153:1162 N-glycan structures 1144:1162 N-glycan structures in O. sativa seedlings 1144:1185 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 3 50 theme biological 300:309 arg1 functions 311:319 N-glycan-mediated biological functions 282:319 N-glycan-mediated biological functions 282:319 However, in plants, N-glycan-mediated biological functions remain obscure. 27320861 4 51 theme complex 441:447 arg1 type 449:452 complex type 441:452 complex type 441:452 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 9 52 from difference 1368:1377 arg1 phenotype 1382:1390 phenotype 1382:1390 phenotype 1382:1390 However, despite a marked difference in phenotype, N-glycan expression in the shoot was also unaffected by light. 27320861 1 53 theme post-translational 100:117 arg1 modifications 119:131 the post-translational modifications 96:131 the post-translational modifications 96:131 N-Glycosylation is one of the post-translational modifications. 27320861 0 54 theme stage 47:51 arg1 transition 9:18 N-glycan transition 0:18 N-glycan transition of the early developmental stage in Oryza sativa 0:67 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 6 55 theme N-glycan 938:945 arg1 structures 947:956 fundamental N-glycan structures 926:956 fundamental N-glycan structures 926:956 The analyses show that fundamental N-glycan structures are common to the shoot and root, whereas paucimannosidic-type N-glycans dramatically decreased in the root grown under light conditions. 27320861 1 56 theme modifications 119:131 arg1 one 89:91 one 89:91 one 89:91 N-Glycosylation is one of the post-translational modifications. 27320861 1 56 theme modifications 119:131 arg1 modifications 119:131 the post-translational modifications 96:131 the post-translational modifications 96:131 N-Glycosylation is one of the post-translational modifications. 27320861 6 57 theme fundamental 926:936 arg1 structures 947:956 fundamental N-glycan structures 926:956 fundamental N-glycan structures 926:956 The analyses show that fundamental N-glycan structures are common to the shoot and root, whereas paucimannosidic-type N-glycans dramatically decreased in the root grown under light conditions. 27320861 5 58 theme N-glycan 688:695 arg1 structure 697:705 N-glycan structure 688:705 N-glycan structure 688:705 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 4 59 theme transition 399:408 arg1 type 382:385 the main type 373:385 the main type of N-glycan transition 373:408 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 5 60 theme N-glycans 755:763 arg1 structures 741:750 the structures 737:750 the structures of N-glycans expressed in O. sativa seedlings grown under light conditions 737:825 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 3 61 theme N-glycan-mediated 282:298 arg1 functions 311:319 N-glycan-mediated biological functions 282:319 N-glycan-mediated biological functions 282:319 However, in plants, N-glycan-mediated biological functions remain obscure. 27320861 11 62 theme light 1683:1687 arg1 conditions 1689:1698 light conditions 1683:1698 light conditions 1683:1698 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 5 63 from analyses 853:860 arg1 root 897:900 root 897:900 root 897:900 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 5 63 from analyses 853:860 arg1 shoot 887:891 shoot 887:891 shoot 887:891 To further elucidate the relationship between N-glycan structure and plant growth, we analyzed the structures of N-glycans expressed in O. sativa seedlings grown under light conditions and performed comparative analyses of the structures in the shoot and root. 27320861 7 64 theme sativa 1226:1231 arg1 seedlings 1233:1241 O. sativa seedlings 1223:1241 O. sativa seedlings grown in the dark 1223:1259 Further, to investigate the effects of light on N-glycan structures in O. sativa seedlings, we analyzed N-glycan structures in O. sativa seedlings grown in the dark. 27320861 2 65 theme cell-cell 187:195 arg1 recognition 197:207 cell-cell recognition 187:207 cell-cell recognition 187:207 In animals, N-glycans linked to proteins function in cell-cell recognition, sorting, transport, and other biological phenomena. 27320861 2 66 theme biological 240:249 arg1 phenomena 251:259 other biological phenomena 234:259 other biological phenomena 234:259 In animals, N-glycans linked to proteins function in cell-cell recognition, sorting, transport, and other biological phenomena. 27320861 10 67 gly glycoproteins 1529:1541 arg1 glycoproteins 1529:1541 different glycoproteins 1519:1541 different glycoproteins 1519:1541 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 11 68 theme many 1709:1712 arg1 proteins 1737:1744 many photosynthesis-related proteins 1709:1744 many photosynthesis-related proteins 1709:1744 Thus, we propose that the N-glycan synthetic system of the O. sativa shoot is almost unaffected by light conditions and that many photosynthesis-related proteins are not modified by N-glycans. 27320861 0 69 theme Oryza 56:60 arg1 sativa 62:67 Oryza sativa 56:67 Oryza sativa 56:67 N-glycan transition of the early developmental stage in Oryza sativa. 27320861 4 70 theme Oryza 486:490 arg1 sativa 492:497 Oryza sativa 486:497 Oryza sativa 486:497 In a previous study, we showed that the main type of N-glycan transition is from the paucimannosidic to complex type before and after germination in Oryza sativa, suggesting that transitions of N-glycan, including those of glycoproteins and glycosyltransferases, are closely associated with plant growth. 27320861 2 71 theme other 234:238 arg1 phenomena 251:259 other biological phenomena 234:259 other biological phenomena 234:259 In animals, N-glycans linked to proteins function in cell-cell recognition, sorting, transport, and other biological phenomena. 27320861 10 72 dep shoot 1486:1490 arg1 the 1482:1484 the 1482:1484 the 1482:1484 This result suggests that the shoot and root of O. sativa have different glycoproteins and distinct N-glycan synthetic systems. 27320861 6 73 theme paucimannosidic-type 1000:1019 arg1 N-glycans 1021:1029 paucimannosidic-type N-glycans 1000:1029 paucimannosidic-type N-glycans 1000:1029 The analyses show that fundamental N-glycan structures are common to the shoot and root, whereas paucimannosidic-type N-glycans dramatically decreased in the root grown under light conditions. 27569415 6 0 theme therapeutic 1389:1399 arg1 protein 1381:1387 a previously uncharacterized non-mAb protein therapeutic 1344:1399 a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines 1344:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 1 1 theme production 272:281 arg1 clone 283:287 final production clone 266:287 final production clone 266:287 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 6 2 theme post-translational 1270:1287 arg1 modifications 1289:1301 unexpected post-translational modifications 1259:1301 unexpected post-translational modifications 1259:1301 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 3 theme non-mAb 1373:1379 arg1 protein 1381:1387 a previously uncharacterized non-mAb protein therapeutic 1344:1399 a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines 1344:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 7 4 theme informed 1533:1540 arg1 selection 1542:1550 a more informed selection 1526:1550 a more informed selection of the production clone 1526:1574 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 2 5 theme early 413:417 arg1 stages 419:424 early stages 413:424 early stages of product development 413:447 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 3 6 theme intense 563:569 arg1 time 580:583 the intense hands-on time 559:583 the intense hands-on time required to generate and analyze LC-MS/MS data 559:630 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 7 7 theme clone 1570:1574 arg1 selection 1542:1550 a more informed selection 1526:1550 a more informed selection of the production clone 1526:1574 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 2 8 theme incomplete 322:331 arg1 understanding 333:345 an incomplete understanding 319:345 an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development 319:447 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 1 9 theme screening 215:223 arg1 methods 225:231 efficient screening methods 205:231 efficient screening methods 205:231 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 6 10 theme host 1450:1453 arg1 lines 1460:1464 three host cell lines 1444:1464 three host cell lines 1444:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 11 theme variants 1332:1339 arg1 glycosylation 1231:1243 glycosylation 1231:1243 glycosylation 1231:1243 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 11 theme variants 1332:1339 arg1 modifications 1289:1301 unexpected post-translational modifications 1259:1301 unexpected post-translational modifications 1259:1301 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 11 theme variants 1332:1339 arg1 types 1314:1318 three types 1308:1318 three types of sequence variants 1308:1339 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 11 theme variants 1332:1339 arg1 degradation 1246:1256 degradation 1246:1256 degradation 1246:1256 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 5 12 theme LC-MS/MS 1146:1153 arg1 results 1155:1161 the LC-MS/MS results 1142:1161 the LC-MS/MS results 1142:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 7 13 theme quality 1483:1489 arg1 risks 1491:1495 Several product quality risks 1467:1495 Several product quality risks 1467:1495 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 3 14 theme LC-MS/MS 618:625 arg1 data 627:630 LC-MS/MS data 618:630 LC-MS/MS data 618:630 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 7 15 theme Several 1467:1473 arg1 risks 1491:1495 Several product quality risks 1467:1495 Several product quality risks 1467:1495 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 4 16 theme peptide 754:760 arg1 mapping 762:768 peptide mapping 754:768 peptide mapping 754:768 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 5 17 theme true-positive 1113:1125 arg1 annotations 1127:1137 true-positive annotations 1113:1137 true-positive annotations 1113:1137 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 3 18 theme sample 668:673 arg1 sets 675:678 only smaller sample sets 655:678 only smaller sample sets 655:678 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 6 19 used used 1185:1188 arg2 methodology 1169:1179 This methodology 1164:1179 This methodology 1164:1179 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 5 20 theme assay 984:988 arg1 control 990:996 an assay control 981:996 an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results 981:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 20 theme assay 984:988 arg1 assessment 1013:1022 an assessment 1010:1022 an assessment of accuracy and precision into the data 1010:1062 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 0 21 theme LC-MS/MS 70:77 arg1 Mapping 87:93 LC-MS/MS Peptide Mapping 70:93 LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein 70:138 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 5 22 theme digestions 943:952 arg1 inclusion 910:918 the inclusion 906:918 the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results 906:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 1 23 theme cell 185:188 arg1 lines 190:194 biopharmaceutical production cell lines 156:194 biopharmaceutical production cell lines 156:194 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 8 24 theme unique 1620:1625 arg1 set 1634:1636 this unique sample set 1615:1636 this unique sample set 1615:1636 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 0 25 theme Clone 99:103 arg1 Selection 105:113 Clone Selection 99:113 Clone Selection of a Therapeutic Protein 99:138 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 8 26 theme important 1647:1655 arg1 information 1676:1686 important structure/function information 1647:1686 important structure/function information 1647:1686 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 3 27 theme peptide 459:465 arg1 mapping 467:473 LC-MS/MS peptide mapping 450:473 LC-MS/MS peptide mapping 450:473 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 0 28 theme Comprehensive 0:12 arg1 Discovery 14:22 Comprehensive Discovery 0:22 Comprehensive Discovery 0:22 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 4 29 theme large 783:787 arg1 sets 796:799 large sample sets 783:799 large sample sets 783:799 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 4 30 theme data 866:869 arg1 analysis 871:878 automated data analysis 856:878 automated data analysis 856:878 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 2 31 theme development 437:447 arg1 stages 419:424 early stages 413:424 early stages of product development 413:447 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 8 32 theme attribute 1707:1715 arg1 identification 1717:1730 quality attribute identification and criticality assessments 1699:1758 identification 1717:1730 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 8 32 theme attribute 1707:1715 arg1 components 1769:1778 two key components 1761:1778 two key components of Quality by Design 1761:1799 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 0 33 theme Heterogeneity 52:64 arg1 Quantitation 28:39 Quantitation 28:39 Quantitation 28:39 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 0 33 theme Heterogeneity 52:64 arg1 Discovery 14:22 Comprehensive Discovery 0:22 Comprehensive Discovery 0:22 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 4 34 theme sample 833:838 arg1 preparation 840:850 higher-throughput sample preparation 815:850 higher-throughput sample preparation 815:850 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 1 35 theme host 247:250 arg1 line 257:260 the host cell line 243:260 the host cell line 243:260 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 1 36 theme biopharmaceutical 156:172 arg1 lines 190:194 biopharmaceutical production cell lines 156:194 biopharmaceutical production cell lines 156:194 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 3 37 theme clone 699:703 arg1 selection 705:713 clone selection 699:713 clone selection 699:713 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 3 38 dep discovery 497:505 arg1 the 493:495 the 493:495 the 493:495 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 3 39 theme heterogeneity 535:547 arg1 quantitation 511:522 quantitation 511:522 quantitation 511:522 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 3 39 theme heterogeneity 535:547 arg1 discovery 497:505 discovery 497:505 discovery 497:505 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 1 40 theme final 266:270 arg1 clone 283:287 final production clone 266:287 final production clone 266:287 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 3 41 theme hands-on 571:578 arg1 time 580:583 the intense hands-on time 559:583 the intense hands-on time required to generate and analyze LC-MS/MS data 559:630 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 6 42 theme unexpected 1259:1268 arg1 modifications 1289:1301 unexpected post-translational modifications 1259:1301 unexpected post-translational modifications 1259:1301 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 8 43 theme Quality 1783:1789 arg1 identification 1717:1730 quality attribute identification and criticality assessments 1699:1758 identification 1717:1730 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 8 43 theme Quality 1783:1789 arg1 components 1769:1778 two key components 1761:1778 two key components of Quality by Design 1761:1799 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 6 44 theme uncharacterized 1357:1371 arg1 protein 1381:1387 a previously uncharacterized non-mAb protein therapeutic 1344:1399 a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines 1344:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 1 45 theme lines 190:194 arg1 Development 141:151 Development 141:151 Development of biopharmaceutical production cell lines 141:194 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 8 46 from set 1634:1636 arg1 inherent 1603:1610 inherent 1603:1610 inherent 1603:1610 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 7 47 theme production 1559:1568 arg1 clone 1570:1574 the production clone 1555:1574 the production clone 1555:1574 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 6 48 gly glycosylation 1231:1243 arg1 protein 1381:1387 a previously uncharacterized non-mAb protein therapeutic 1344:1399 a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines 1344:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 48 gly glycosylation 1231:1243 arg1 variants 1332:1339 sequence variants 1323:1339 sequence variants 1323:1339 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 1 49 theme efficient 205:213 arg1 methods 225:231 efficient screening methods 205:231 efficient screening methods 205:231 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 0 50 theme Protein 132:138 arg1 Selection 105:113 Clone Selection 99:113 Clone Selection of a Therapeutic Protein 99:138 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 5 51 theme multiple 958:965 arg1 inclusion 910:918 the inclusion 906:918 the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results 906:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 2 52 theme relationship 354:365 arg1 understanding 333:345 an incomplete understanding 319:345 an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development 319:447 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 5 53 theme orthogonal 923:932 arg1 digestions 943:952 orthogonal protease digestions 923:952 orthogonal protease digestions 923:952 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 6 54 theme sequence 1323:1330 arg1 variants 1332:1339 sequence variants 1323:1339 sequence variants 1323:1339 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 6 55 theme cell 1455:1458 arg1 lines 1460:1464 three host cell lines 1444:1464 three host cell lines 1444:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 4 56 theme simple 735:740 arg1 approach 742:749 a simple approach 733:749 a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis 733:878 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 3 57 theme smaller 660:666 arg1 sets 675:678 only smaller sample sets 655:678 only smaller sample sets 655:678 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 8 58 theme inherent 1603:1610 arg1 variability 1591:1601 the variability 1587:1601 the variability inherent in this unique sample set 1587:1636 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 7 59 theme product 1475:1481 arg1 risks 1491:1495 Several product quality risks 1467:1495 Several product quality risks 1467:1495 Several product quality risks were identified allowing for a more informed selection of the production clone. 27569415 0 60 theme Peptide 79:85 arg1 Mapping 87:93 LC-MS/MS Peptide Mapping 70:93 LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein 70:138 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 5 61 theme annotations 1127:1137 arg1 identification 1095:1108 the identification 1091:1108 the identification of true-positive annotations in the LC-MS/MS results 1091:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 62 theme protease 934:941 arg1 digestions 943:952 orthogonal protease digestions 923:952 orthogonal protease digestions 923:952 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 63 from identification 1095:1108 arg1 results 1155:1161 the LC-MS/MS results 1142:1161 the LC-MS/MS results 1142:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 1 64 theme production 174:183 arg1 lines 190:194 biopharmaceutical production cell lines 156:194 biopharmaceutical production cell lines 156:194 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 0 65 theme Therapeutic 120:130 arg1 Protein 132:138 a Therapeutic Protein 118:138 a Therapeutic Protein 118:138 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 8 66 theme sample 1627:1632 arg1 set 1634:1636 this unique sample set 1615:1636 this unique sample set 1615:1636 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 8 67 from inherent 1603:1610 arg1 set 1634:1636 this unique sample set 1615:1636 this unique sample set 1615:1636 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 2 68 dep protein 375:381 arg1 heterogeneity 383:395 heterogeneity 383:395 heterogeneity 383:395 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 4 69 theme sample 789:794 arg1 sets 796:799 large sample sets 783:799 large sample sets 783:799 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 6 70 from lines 1460:1464 arg1 clones 1432:1437 approximately 100 clones 1414:1437 approximately 100 clones from three host cell lines 1414:1464 This methodology was used to comprehensively identify and quantify glycosylation, degradation, unexpected post-translational modifications, and three types of sequence variants in a previously uncharacterized non-mAb protein therapeutic expressed in approximately 100 clones from three host cell lines. 27569415 5 71 theme accuracy 1027:1034 arg1 control 990:996 an assay control 981:996 an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results 981:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 71 theme accuracy 1027:1034 arg1 assessment 1013:1022 an assessment 1010:1022 an assessment of accuracy and precision into the data 1010:1062 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 8 72 theme structure/function 1657:1674 arg1 information 1676:1686 important structure/function information 1647:1686 important structure/function information 1647:1686 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 5 73 theme control 990:996 arg1 digestions 943:952 orthogonal protease digestions 923:952 orthogonal protease digestions 923:952 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 74 dep multiple 958:965 arg1 replicates 967:976 replicates 967:976 replicates 967:976 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 8 75 theme quality 1699:1705 arg1 identification 1717:1730 quality attribute identification and criticality assessments 1699:1758 identification 1717:1730 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 8 75 theme quality 1699:1705 arg1 components 1769:1778 two key components 1761:1778 two key components of Quality by Design 1761:1799 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 3 76 theme later 683:687 arg1 stages 689:694 later stages 683:694 later stages of clone selection 683:713 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 0 77 theme Protein 44:50 arg1 Heterogeneity 52:64 Protein Heterogeneity 44:64 Protein Heterogeneity 44:64 Comprehensive Discovery and Quantitation of Protein Heterogeneity via LC-MS/MS Peptide Mapping for Clone Selection of a Therapeutic Protein. 27569415 3 78 theme LC-MS/MS 450:457 arg1 mapping 467:473 LC-MS/MS peptide mapping 450:473 LC-MS/MS peptide mapping 450:473 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 4 79 theme automated 856:864 arg1 analysis 871:878 automated data analysis 856:878 automated data analysis 856:878 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 2 80 theme product 429:435 arg1 development 437:447 product development 429:447 product development 429:447 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 8 81 theme criticality 1736:1746 arg1 assessments 1748:1758 quality attribute identification and criticality assessments 1699:1758 assessments 1748:1758 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 3 82 theme selection 705:713 arg1 stages 689:694 later stages 683:694 later stages of clone selection 683:713 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 3 83 theme protein 527:533 arg1 heterogeneity 535:547 protein heterogeneity 527:547 protein heterogeneity 527:547 LC-MS/MS peptide mapping is well suited to the discovery and quantitation of protein heterogeneity; however, the intense hands-on time required to generate and analyze LC-MS/MS data typically accommodates only smaller sample sets at later stages of clone selection. 27569415 5 84 theme precision 1040:1048 arg1 control 990:996 an assay control 981:996 an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results 981:1161 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 5 84 theme precision 1040:1048 arg1 assessment 1013:1022 an assessment 1010:1022 an assessment of accuracy and precision into the data 1010:1062 This approach allows for the inclusion of orthogonal protease digestions and multiple replicates of an assay control that encode an assessment of accuracy and precision into the data, significantly simplifying the identification of true-positive annotations in the LC-MS/MS results. 27569415 2 85 from stages 419:424 arg1 protein 375:381 protein heterogeneity and function 375:408 protein 375:381 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 2 85 from stages 419:424 arg1 function 401:408 function 401:408 function 401:408 This is often complicated by an incomplete understanding of the relationship between protein heterogeneity and function at early stages of product development. 27569415 4 86 theme higher-throughput 815:831 arg1 preparation 840:850 higher-throughput sample preparation 815:850 higher-throughput sample preparation 815:850 Here we describe a simple approach to peptide mapping designed for large sample sets that includes higher-throughput sample preparation and automated data analysis. 27569415 1 87 theme cell 252:255 arg1 line 257:260 the host cell line 243:260 the host cell line 243:260 Development of biopharmaceutical production cell lines requires efficient screening methods to select the host cell line and final production clone. 27569415 8 88 theme key 1765:1767 arg1 identification 1717:1730 quality attribute identification and criticality assessments 1699:1758 identification 1717:1730 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 27569415 8 88 theme key 1765:1767 arg1 components 1769:1778 two key components 1761:1778 two key components of Quality by Design 1761:1799 Moreover, the variability inherent in this unique sample set provides important structure/function information to support quality attribute identification and criticality assessments, two key components of Quality by Design. 28482115 0 0 theme carboxy-proximal 82:97 arg1 domain 111:116 its carboxy-proximal Fasciclin 1 domain 78:116 its carboxy-proximal Fasciclin 1 domain 78:116 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 4 1 theme sites 889:893 arg1 N-glycan 835:842 a highly conserved N-glycan 816:842 a highly conserved N-glycan 816:842 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 4 1 theme sites 889:893 arg1 signal 808:813 the GPI-modification signal 787:813 the GPI-modification signal 787:813 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 4 1 theme sites 889:893 arg1 deletion 851:858 the deletion 847:858 the deletion of predicted O-glycosylation sites 847:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 2 2 theme Arabidopsis 373:383 arg1 FLA4 394:397 Arabidopsis thaliana FLA4 373:397 Arabidopsis thaliana FLA4 373:397 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 1 3 theme structure 256:264 arg1 relevance 229:237 the relevance 225:237 the relevance of their complex structure to physiological function and cellular fate 225:308 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 1 3 theme structure 256:264 arg1 unresolved 313:322 unresolved 313:322 unresolved 313:322 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 5 4 theme endoplasmic 956:966 arg1 ER 979:980 ER 979:980 ER 979:980 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 5 4 theme endoplasmic 956:966 arg1 reticulum 968:976 endoplasmic reticulum 956:976 endoplasmic reticulum (ER)-exit 956:986 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 4 5 theme O-glycosylation 873:887 arg1 sites 889:893 predicted O-glycosylation sites 863:893 predicted O-glycosylation sites 863:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 6 6 theme amino-proximal 1261:1274 arg1 domain 1288:1293 its amino-proximal fasciclin 1 domain 1257:1293 its amino-proximal fasciclin 1 domain 1257:1293 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 6 7 theme localization 1344:1355 arg1 stabilization 1311:1323 stabilization 1311:1323 stabilization of plasma membrane localization 1311:1355 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 2 8 dep Arabidopsis 373:383 arg1 thaliana 385:392 thaliana 385:392 thaliana 385:392 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 1 9 theme physiological 269:281 arg1 function 283:290 physiological function 269:290 physiological function 269:290 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 0 10 theme Fasciclin 99:107 arg1 domain 111:116 its carboxy-proximal Fasciclin 1 domain 78:116 its carboxy-proximal Fasciclin 1 domain 78:116 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 6 11 theme membrane 1335:1342 arg1 localization 1344:1355 plasma membrane localization 1328:1355 plasma membrane localization 1328:1355 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 5 12 theme membrane 999:1006 arg1 localization 1008:1019 plasma membrane localization 992:1019 plasma membrane localization of FLA4 992:1027 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 3 13 gly N-glycosylated 559:572 arg1 FLA4 514:517 FLA4 514:517 FLA4 514:517 FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. 28482115 4 14 theme deletion 851:858 arg1 removal 776:782 removal 776:782 removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites 776:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 5 15 theme reticulum 968:976 arg1 -exit 982:986 endoplasmic reticulum (ER)-exit 956:986 endoplasmic reticulum (ER)-exit 956:986 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 4 16 gly O-glycosylation 873:887 arg2 sites 889:893 predicted O-glycosylation sites 863:893 predicted O-glycosylation sites 863:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 3 17 contain carries 578:584 arg1 FLA4 514:517 FLA4 514:517 FLA4 514:517 FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. 28482115 3 17 contain carries 578:584 arg2 epitopes 599:606 two O-glycan epitopes 586:606 two O-glycan epitopes previously associated with arabinogalactan proteins 586:658 FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. 28482115 6 18 theme fasciclin 1229:1237 arg1 domain 1241:1246 its carboxy-proximal fasciclin 1 domain 1208:1246 its carboxy-proximal fasciclin 1 domain 1208:1246 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 1 19 theme numerous 182:189 arg1 functions 201:209 numerous important functions 182:209 numerous important functions in plants 182:219 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 0 20 theme thaliana 12:19 arg1 FLA4 21:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 0 20 theme thaliana 12:19 arg1 factor 67:72 a glycan-stabilized soluble factor 39:72 a glycan-stabilized soluble factor 39:72 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 4 21 theme predicted 863:871 arg1 sites 889:893 predicted O-glycosylation sites 863:893 predicted O-glycosylation sites 863:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 6 22 theme carboxy-proximal 1212:1227 arg1 domain 1241:1246 its carboxy-proximal fasciclin 1 domain 1208:1246 its carboxy-proximal fasciclin 1 domain 1208:1246 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 1 23 theme important 191:199 arg1 functions 201:209 numerous important functions 182:209 numerous important functions in plants 182:219 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 0 24 theme Arabidopsis 0:10 arg1 thaliana 12:19 Arabidopsis thaliana 0:19 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 6 25 theme plasma 1328:1333 arg1 localization 1344:1355 plasma membrane localization 1328:1355 plasma membrane localization 1328:1355 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 7 26 gly glycoprotein 1386:1397 arg1 glycoprotein 1386:1397 a soluble glycoprotein 1376:1397 a soluble glycoprotein 1376:1397 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 7 26 gly glycoprotein 1386:1397 arg1 FLA4 1358:1361 FLA4 1358:1361 FLA4 1358:1361 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 4 27 theme domain 747:752 arg1 deletion 704:711 deletion 704:711 deletion of the amino-proximal fasciclin 1 domain 704:752 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 4 28 theme FLA4 677:680 arg1 resistant 686:694 resistant 686:694 resistant 686:694 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 4 28 theme FLA4 677:680 arg1 activity 665:672 The activity 661:672 The activity of FLA4 661:680 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 2 29 theme plasma 445:450 arg1 membrane 452:459 the plasma membrane 441:459 the plasma membrane 441:459 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 1 30 theme cellular 296:303 arg1 fate 305:308 cellular fate 296:308 cellular fate 296:308 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 7 31 theme cellular 1451:1458 arg1 trafficking 1460:1470 its normal cellular trafficking 1440:1470 its normal cellular trafficking 1440:1470 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 5 32 theme plasma 992:997 arg1 localization 1008:1019 plasma membrane localization 992:1019 plasma membrane localization of FLA4 992:1027 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 2 33 from soluble 489:495 arg1 apoplast 504:511 the apoplast 500:511 the apoplast 500:511 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 7 34 theme soluble 1378:1384 arg1 glycoprotein 1386:1397 a soluble glycoprotein 1376:1397 a soluble glycoprotein 1376:1397 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 7 34 theme soluble 1378:1384 arg1 FLA4 1358:1361 FLA4 1358:1361 FLA4 1358:1361 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 2 35 theme fluorescent 350:360 arg1 version 362:368 a fully functional fluorescent version 331:368 a fully functional fluorescent version of Arabidopsis thaliana FLA4 331:397 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 5 36 theme post-secretory 1114:1127 arg1 fate 1129:1132 post-secretory fate 1114:1132 post-secretory fate 1114:1132 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 4 37 theme conserved 825:833 arg1 N-glycan 835:842 a highly conserved N-glycan 816:842 a highly conserved N-glycan 816:842 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 3 38 theme arabinogalactan 635:649 arg1 proteins 651:658 arabinogalactan proteins 635:658 arabinogalactan proteins 635:658 FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. 28482115 3 39 theme O-glycan 590:597 arg1 epitopes 599:606 two O-glycan epitopes 586:606 two O-glycan epitopes previously associated with arabinogalactan proteins 586:658 FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with arabinogalactan proteins. 28482115 0 40 theme soluble 59:65 arg1 FLA4 21:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 0 40 theme soluble 59:65 arg1 factor 67:72 a glycan-stabilized soluble factor 39:72 a glycan-stabilized soluble factor 39:72 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 5 41 theme ER-exit 1074:1080 arg1 level 1065:1069 the level 1061:1069 the level of ER-exit and O-glycosylation influencing post-secretory fate 1061:1132 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 7 42 theme normal 1444:1449 arg1 trafficking 1460:1470 its normal cellular trafficking 1440:1470 its normal cellular trafficking 1440:1470 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 1 43 theme Fasciclin-like 119:132 arg1 proteins 150:157 Fasciclin-like arabinogalactan proteins 119:157 Fasciclin-like arabinogalactan proteins (FLAs) 119:164 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 1 43 theme Fasciclin-like 119:132 arg1 FLAs 160:163 FLAs 160:163 FLAs 160:163 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 0 44 theme glycan-stabilized 41:57 arg1 FLA4 21:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 0:24 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 0 44 theme glycan-stabilized 41:57 arg1 factor 67:72 a glycan-stabilized soluble factor 39:72 a glycan-stabilized soluble factor 39:72 Arabidopsis thaliana FLA4 functions as a glycan-stabilized soluble factor via its carboxy-proximal Fasciclin 1 domain. 28482115 4 45 theme N-glycan 835:842 arg1 removal 776:782 removal 776:782 removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites 776:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 1 46 theme arabinogalactan 134:148 arg1 proteins 150:157 Fasciclin-like arabinogalactan proteins 119:157 Fasciclin-like arabinogalactan proteins (FLAs) 119:164 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 1 46 theme arabinogalactan 134:148 arg1 FLAs 160:163 FLAs 160:163 FLAs 160:163 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 4 47 theme amino-proximal 720:733 arg1 domain 747:752 the amino-proximal fasciclin 1 domain 716:752 the amino-proximal fasciclin 1 domain 716:752 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 2 48 from apoplast 504:511 arg1 soluble 489:495 soluble 489:495 soluble 489:495 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 2 49 dep show 402:405 arg1 Using 325:329 Using 325:329 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 325:397 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 2 50 theme functional 339:348 arg1 version 362:368 a fully functional fluorescent version 331:368 a fully functional fluorescent version of Arabidopsis thaliana FLA4 331:397 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 6 51 theme fasciclin 1276:1284 arg1 domain 1288:1293 its amino-proximal fasciclin 1 domain 1257:1293 its amino-proximal fasciclin 1 domain 1257:1293 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 4 52 theme GPI-modification 791:806 arg1 signal 808:813 the GPI-modification signal 787:813 the GPI-modification signal 787:813 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 4 53 theme fasciclin 735:743 arg1 domain 747:752 the amino-proximal fasciclin 1 domain 716:752 the amino-proximal fasciclin 1 domain 716:752 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 6 54 theme molecular 1175:1183 arg1 interactions 1185:1196 molecular interactions 1175:1196 molecular interactions involving its carboxy-proximal fasciclin 1 domain 1175:1246 We show that FLA4 acts predominantly by molecular interactions involving its carboxy-proximal fasciclin 1 domain and that its amino-proximal fasciclin 1 domain is required for stabilization of plasma membrane localization. 28482115 4 55 theme signal 808:813 arg1 removal 776:782 removal 776:782 removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites 776:893 The activity of FLA4 was resistant against deletion of the amino-proximal fasciclin 1 domain and was unaffected by removal of the GPI-modification signal, a highly conserved N-glycan or the deletion of predicted O-glycosylation sites. 28482115 5 56 theme O-glycosylation 1086:1100 arg1 level 1065:1069 the level 1061:1069 the level of ER-exit and O-glycosylation influencing post-secretory fate 1061:1132 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 7 57 theme Fas1 1424:1427 arg1 domain 1429:1434 its carboxy-proximal Fas1 domain 1403:1434 its carboxy-proximal Fas1 domain 1403:1434 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 1 58 from functions 201:209 arg1 plants 214:219 plants 214:219 plants 214:219 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 28482115 2 59 theme FLA4 394:397 arg1 version 362:368 a fully functional fluorescent version 331:368 a fully functional fluorescent version of Arabidopsis thaliana FLA4 331:397 Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. 28482115 7 60 theme carboxy-proximal 1407:1422 arg1 domain 1429:1434 its carboxy-proximal Fas1 domain 1403:1434 its carboxy-proximal Fas1 domain 1403:1434 FLA4 functions as a soluble glycoprotein via its carboxy-proximal Fas1 domain and its normal cellular trafficking depends on N- and O-glycosylation. 28482115 5 61 theme FLA4 1024:1027 arg1 -exit 982:986 endoplasmic reticulum (ER)-exit 956:986 endoplasmic reticulum (ER)-exit 956:986 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 5 61 theme FLA4 1024:1027 arg1 localization 1008:1019 plasma membrane localization 992:1019 plasma membrane localization of FLA4 992:1027 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 5 62 theme structural 914:923 arg1 changes 925:931 these structural changes 908:931 these structural changes 908:931 Nonetheless these structural changes dramatically decreased endoplasmic reticulum (ER)-exit and plasma membrane localization of FLA4, with N-glycosylation acting at the level of ER-exit and O-glycosylation influencing post-secretory fate. 28482115 1 63 theme complex 248:254 arg1 structure 256:264 their complex structure 242:264 their complex structure 242:264 Fasciclin-like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. 26335373 4 0 theme PNGase 613:618 arg1 F 620:620 PNGase F 613:620 PNGase F 613:620 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 4 0 theme PNGase 613:618 arg1 F 610:610 peptide N-glycosidase F 588:610 peptide N-glycosidase F (PNGase F) 588:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 4 1 theme prior 555:559 arg1 glycans 547:553 corresponding glycans 533:553 corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F) 533:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 7 2 theme glycoproteome 1133:1145 arg1 analysis 1108:1115 detailed analysis 1099:1115 detailed analysis of O-glycome or glycoproteome 1099:1145 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 5 3 theme Non-bound 624:632 arg1 glycans 634:640 Non-bound glycans 624:640 Non-bound glycans 624:640 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 2 4 theme improved 286:293 arg1 depth-of-coverage 295:311 improved depth-of-coverage 286:311 improved depth-of-coverage 286:311 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 5 5 theme matrix-assisted 689:703 arg1 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 5 5 theme matrix-assisted 689:703 arg1 spectrometry 753:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 689:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 689:779 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 6 6 theme mammary 859:865 arg1 NMuMG 885:889 NMuMG 885:889 NMuMG 885:889 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 6 6 theme mammary 859:865 arg1 cells 878:882 normal mouse mammary epithelial cells 846:882 normal mouse mammary epithelial cells (NMuMG) 846:890 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 7 7 theme biological 1056:1065 arg1 samples 1067:1073 biological samples 1056:1073 biological samples 1056:1073 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 5 8 theme bound 646:650 arg1 glycans 652:658 bound glycans 646:658 bound glycans 646:658 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 6 9 theme mouse 853:857 arg1 NMuMG 885:889 NMuMG 885:889 NMuMG 885:889 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 6 9 theme mouse 853:857 arg1 cells 878:882 normal mouse mammary epithelial cells 846:882 normal mouse mammary epithelial cells (NMuMG) 846:890 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 2 10 theme glycans 277:283 arg1 complexity 245:254 general complexity 237:254 general complexity 237:254 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 2 10 theme glycans 277:283 arg1 heterogeneity 260:272 heterogeneity 260:272 heterogeneity 260:272 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 7 11 from identification 1018:1031 arg1 samples 1067:1073 biological samples 1056:1073 biological samples 1056:1073 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 2 12 dep complexity 245:254 arg1 the 233:235 the 233:235 the 233:235 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 6 13 theme normal 846:851 arg1 NMuMG 885:889 NMuMG 885:889 NMuMG 885:889 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 6 13 theme normal 846:851 arg1 cells 878:882 normal mouse mammary epithelial cells 846:882 normal mouse mammary epithelial cells (NMuMG) 846:890 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 1 14 dep tool 140:143 arg1 elucidated 210:219 elucidated 210:219 are elucidated 206:219 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 6 15 theme additional 947:956 arg1 structures 967:976 5, 6, and 11 additional N-glycan structures 934:976 5, 6, and 11 additional N-glycan structures 934:976 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 0 16 theme lectin-based 2:13 arg1 strategy 36:43 A lectin-based isolation/enrichment strategy 0:43 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis 0:86 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis. 26335373 3 17 theme total 451:455 arg1 information 466:476 total glycomic information 451:476 total glycomic information 451:476 In this study, we established the lectin-based isolation/enrichment strategy for total glycomic information. 26335373 2 18 gly heterogeneity 260:272 arg1 glycans 277:283 glycans 277:283 glycans 277:283 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 4 19 theme peptide 588:594 arg1 F 620:620 PNGase F 613:620 PNGase F 613:620 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 4 19 theme peptide 588:594 arg1 F 610:610 peptide N-glycosidase F 588:610 peptide N-glycosidase F (PNGase F) 588:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 7 20 theme O-glycome 1120:1128 arg1 analysis 1108:1115 detailed analysis 1099:1115 detailed analysis of O-glycome or glycoproteome 1099:1145 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 1 21 theme animal 191:196 arg1 species 198:204 more and more animal species 177:204 more and more animal species 177:204 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 6 22 theme chicken 827:833 arg1 ovalbumin 835:843 chicken ovalbumin 827:843 chicken ovalbumin 827:843 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 1 23 dep genomes 152:158 arg1 the 148:150 the 148:150 the 148:150 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 1 24 theme species 198:204 arg1 proteomes 164:172 proteomes 164:172 proteomes 164:172 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 1 24 theme species 198:204 arg1 genomes 152:158 genomes 152:158 genomes 152:158 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 0 25 theme isolation/enrichment 15:34 arg1 strategy 36:43 A lectin-based isolation/enrichment strategy 0:43 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis 0:86 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis. 26335373 7 26 theme detailed 1099:1106 arg1 analysis 1108:1115 detailed analysis 1099:1115 detailed analysis of O-glycome or glycoproteome 1099:1145 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 1 27 theme useful 124:129 arg1 tool 140:143 an increasingly useful research tool 108:143 an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated 108:219 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 0 28 theme improved 49:56 arg1 coverage 58:65 improved coverage 49:65 improved coverage of N-glycan analysis 49:86 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis. 26335373 4 29 theme N-glycosidase 596:608 arg1 F 620:620 PNGase F 613:620 PNGase F 613:620 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 4 29 theme N-glycosidase 596:608 arg1 F 610:610 peptide N-glycosidase F 588:610 peptide N-glycosidase F (PNGase F) 588:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 4 30 theme Specific 479:486 arg1 lectins 488:494 Specific lectins 479:494 Specific lectins 479:494 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 3 31 theme glycomic 457:464 arg1 information 466:476 total glycomic information 451:476 total glycomic information 451:476 In this study, we established the lectin-based isolation/enrichment strategy for total glycomic information. 26335373 6 32 theme structures 967:976 arg1 detection 921:929 detection 921:929 detection of 5, 6, and 11 additional N-glycan structures 921:976 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 6 33 theme human 897:901 arg1 serum 903:907 human serum 897:907 human serum 897:907 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 3 34 theme lectin-based 404:415 arg1 strategy 438:445 the lectin-based isolation/enrichment strategy 400:445 the lectin-based isolation/enrichment strategy for total glycomic information 400:476 In this study, we established the lectin-based isolation/enrichment strategy for total glycomic information. 26335373 6 35 theme N-glycan 958:965 arg1 structures 967:976 5, 6, and 11 additional N-glycan structures 934:976 5, 6, and 11 additional N-glycan structures 934:976 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 5 36 theme laser 705:709 arg1 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 5 36 theme laser 705:709 arg1 spectrometry 753:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 689:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 689:779 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 6 37 theme strategy 815:822 arg1 Application 796:806 Application 796:806 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum 796:907 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 1 38 theme research 131:138 arg1 tool 140:143 an increasingly useful research tool 108:143 an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated 108:219 Glycomics provides an increasingly useful research tool as the genomes and proteomes of more and more animal species are elucidated. 26335373 3 39 theme isolation/enrichment 417:436 arg1 strategy 438:445 the lectin-based isolation/enrichment strategy 400:445 the lectin-based isolation/enrichment strategy for total glycomic information 400:476 In this study, we established the lectin-based isolation/enrichment strategy for total glycomic information. 26335373 4 40 theme N-glycans 575:583 arg1 release 564:570 release 564:570 release of N-glycans by peptide N-glycosidase F (PNGase F) 564:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 5 41 theme desorption/ionization 711:731 arg1 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 5 41 theme desorption/ionization 711:731 arg1 spectrometry 753:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 689:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 689:779 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 7 42 theme intact 1036:1041 arg1 N-glycans 1043:1051 intact N-glycans 1036:1051 intact N-glycans 1036:1051 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 2 43 theme glycosylation 346:358 arg1 analysis 360:367 glycosylation analysis 346:367 glycosylation analysis 346:367 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 5 44 theme time-of-flight 733:746 arg1 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 5 44 theme time-of-flight 733:746 arg1 spectrometry 753:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 689:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 689:779 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 2 45 theme heterogeneity 260:272 arg1 view 225:228 view 225:228 view of the general complexity and heterogeneity of glycans 225:283 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 6 46 theme epithelial 867:876 arg1 NMuMG 885:889 NMuMG 885:889 NMuMG 885:889 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 6 46 theme epithelial 867:876 arg1 cells 878:882 normal mouse mammary epithelial cells 846:882 normal mouse mammary epithelial cells (NMuMG) 846:890 Application of the strategy to chicken ovalbumin, normal mouse mammary epithelial cells (NMuMG), and human serum resulted in detection of 5, 6, and 11 additional N-glycan structures, respectively. 26335373 5 47 theme mass 748:751 arg1 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 MALDI-TOF-MS 767:778 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 5 47 theme mass 748:751 arg1 spectrometry 753:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 689:764 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 689:779 Non-bound glycans and bound glycans are released and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), respectively. 26335373 0 48 theme analysis 79:86 arg1 coverage 58:65 improved coverage 49:65 improved coverage of N-glycan analysis 49:86 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis. 26335373 7 49 theme N-glycans 1043:1051 arg1 identification 1018:1031 identification 1018:1031 identification of intact N-glycans in biological samples 1018:1073 The strategy facilitates identification of intact N-glycans in biological samples, and can be extended to detailed analysis of O-glycome or glycoproteome. 26335373 2 50 theme complexity 245:254 arg1 view 225:228 view 225:228 view of the general complexity and heterogeneity of glycans 225:283 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 26335373 0 51 theme N-glycan 70:77 arg1 analysis 79:86 N-glycan analysis 70:86 N-glycan analysis 70:86 A lectin-based isolation/enrichment strategy for improved coverage of N-glycan analysis. 26335373 4 52 theme corresponding 533:545 arg1 glycans 547:553 corresponding glycans 533:553 corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F) 533:621 Specific lectins are added onto the filter to capture corresponding glycans prior to release of N-glycans by peptide N-glycosidase F (PNGase F). 26335373 2 53 theme general 237:243 arg1 complexity 245:254 general complexity 237:254 general complexity 237:254 In view of the general complexity and heterogeneity of glycans, improved depth-of-coverage and sensitivity are required for glycosylation analysis. 28735452 9 0 theme fractogram 1906:1915 arg1 components 1885:1894 the components 1881:1894 the components of an AF4 fractogram 1881:1915 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 9 1 dep design 1948:1953 arg1 the 1935:1937 the 1935:1937 the 1935:1937 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 2 2 theme polymers 311:318 arg1 mass 303:306 The molar mass 293:306 The molar mass of polymers and their degradation products 293:349 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 1 3 theme various 206:212 arg1 polyphenols 280:290 polyphenols 280:290 polyphenols 280:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 3 theme various 206:212 arg1 polysaccharides 249:263 polysaccharides 249:263 polysaccharides 249:263 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 3 theme various 206:212 arg1 macromolecules 224:237 various polymeric macromolecules 206:237 various polymeric macromolecules including polysaccharides, peptides, and polyphenols 206:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 3 theme various 206:212 arg1 peptides 266:273 peptides 266:273 peptides 266:273 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 7 4 theme non-starch 1504:1513 arg1 polysaccharides 1515:1529 non-starch polysaccharides 1504:1529 non-starch polysaccharides 1504:1529 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 4 5 theme cereal-based 897:908 arg1 beverage 910:917 a yeast-fermented cereal-based beverage 879:917 a yeast-fermented cereal-based beverage 879:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 4 5 theme cereal-based 897:908 arg1 beer 873:876 beer 873:876 beer 873:876 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 2 6 theme different 358:366 arg1 parameters 405:414 different technological and especially sensory parameters 358:414 different technological and especially sensory parameters of beverages 358:427 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 8 7 theme starch 1768:1773 arg1 hydrolysis 1775:1784 incomplete starch hydrolysis 1757:1784 incomplete starch hydrolysis 1757:1784 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 1 8 theme macromolecules 224:237 arg1 mixture 195:201 a complex mixture 185:201 a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols 185:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 9 9 from evaluation 1959:1968 arg1 beverages 2008:2016 fermented cereal-based beverages 1985:2016 fermented cereal-based beverages 1985:2016 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 7 10 with molecules 1351:1359 arg1 degrees 1376:1382 different degrees 1366:1382 different degrees of glycosylation 1366:1399 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 2 11 theme products 342:349 arg1 mass 303:306 The molar mass 293:306 The molar mass of polymers and their degradation products 293:349 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 7 12 theme mass 1423:1426 arg1 fraction 1428:1435 the middle molar mass fraction 1406:1435 the middle molar mass fraction 1406:1435 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 13 theme polymers 1973:1980 arg1 evaluation 1959:1968 evaluation 1959:1968 evaluation 1959:1968 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 9 13 theme polymers 1973:1980 arg1 design 1948:1953 design 1948:1953 design 1948:1953 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 3 14 theme distribution 630:641 arg1 analysis 643:650 molar mass distribution analysis 619:650 molar mass distribution analysis 619:650 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 7 15 theme middle 1410:1415 arg1 fraction 1428:1435 the middle molar mass fraction 1406:1435 the middle molar mass fraction 1406:1435 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 16 theme fermented 1985:1993 arg1 beverages 2008:2016 fermented cereal-based beverages 1985:2016 fermented cereal-based beverages 1985:2016 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 5 17 with hydrolysis 1035:1044 arg1 standards 1084:1092 purified polymer standards 1067:1092 purified polymer standards 1067:1092 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 3 18 theme molar 619:623 arg1 analysis 643:650 molar mass distribution analysis 619:650 molar mass distribution analysis 619:650 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 4 19 theme systematic 758:767 arg1 approach 769:776 a systematic approach 756:776 a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage 756:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 6 20 theme Corresponding 1095:1107 arg1 effects 1109:1115 Corresponding effects 1095:1115 Corresponding effects on dRI and UV signals 1095:1137 Corresponding effects on dRI and UV signals were evaluated according to the detector's sensitivities. 28735452 4 21 from fractogram 837:846 arg1 beverage 910:917 a yeast-fermented cereal-based beverage 879:917 a yeast-fermented cereal-based beverage 879:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 4 21 from fractogram 837:846 arg1 beer 873:876 beer 873:876 beer 873:876 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 5 22 theme fractions 945:953 arg1 Assignment 920:929 Assignment 920:929 Assignment of fractogram fractions to polymer substance classes 920:982 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 7 23 theme wall 1609:1612 arg1 polysaccharides 1614:1628 the cell wall polysaccharides 1600:1628 the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) 1600:1662 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 0 24 theme flow 93:96 arg1 fractionation 109:121 asymmetrical flow field-flow fractionation 80:121 asymmetrical flow field-flow fractionation with a multi-detection system 80:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 8 25 theme mass 1847:1850 arg1 range 1852:1856 the complete molar mass range 1828:1856 the complete molar mass range 1828:1856 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 5 26 theme polymer 958:964 arg1 classes 976:982 polymer substance classes 958:982 polymer substance classes 958:982 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 0 27 from Characterization 0:15 arg1 beverages 64:72 cereal-based beverages 51:72 cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system 51:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 6 28 from effects 1109:1115 arg1 UV 1128:1129 UV 1128:1129 UV 1128:1129 Corresponding effects on dRI and UV signals were evaluated according to the detector's sensitivities. 28735452 6 28 from effects 1109:1115 arg1 dRI 1120:1122 dRI 1120:1122 dRI 1120:1122 Corresponding effects on dRI and UV signals were evaluated according to the detector's sensitivities. 28735452 9 29 theme quality 2053:2059 arg1 parameters 2061:2070 quality parameters 2053:2070 quality parameters 2053:2070 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 6 30 dep dRI 1120:1122 arg1 signals 1131:1137 signals 1131:1137 signals 1131:1137 Corresponding effects on dRI and UV signals were evaluated according to the detector's sensitivities. 28735452 4 31 from classes 806:812 arg1 fractogram 837:846 an AF4//MALS/dRI/UV fractogram 817:846 an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage 817:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 9 32 from beverages 2008:2016 arg1 evaluation 1959:1968 evaluation 1959:1968 evaluation 1959:1968 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 9 32 from beverages 2008:2016 arg1 design 1948:1953 design 1948:1953 design 1948:1953 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 7 33 dep β-glucan 1637:1644 arg1 i.e. 1631:1634 i.e. 1631:1634 i.e. 1631:1634 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 34 theme association 1709:1719 arg1 content 1675:1681 a low content 1669:1681 a low content of polysaccharide-protein association 1669:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 35 theme multi-angle 492:502 arg1 scattering 510:519 multi-angle light scattering 492:519 multi-angle light scattering (MALS) 492:526 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 35 theme multi-angle 492:502 arg1 dRI 560:562 dRI 560:562 dRI 560:562 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 35 theme multi-angle 492:502 arg1 MALS 522:525 MALS 522:525 MALS 522:525 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 5 36 theme targeted 1000:1007 arg1 precipitations 1009:1022 targeted precipitations 1000:1022 targeted precipitations 1000:1022 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 3 37 theme flow 443:446 arg1 fractionation 459:471 Asymmetrical flow field-flow fractionation 430:471 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) 430:584 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 37 theme flow 443:446 arg1 technique 591:599 a technique 589:599 a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions 589:710 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 37 theme flow 443:446 arg1 AF4 474:476 AF4 474:476 AF4 474:476 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 0 38 theme polymeric 20:28 arg1 classes 40:46 polymeric substance classes 20:46 polymeric substance classes 20:46 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 7 39 theme low 1671:1673 arg1 content 1675:1681 a low content 1669:1681 a low content of polysaccharide-protein association 1669:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 40 theme different 1366:1374 arg1 degrees 1376:1382 different degrees 1366:1382 different degrees of glycosylation 1366:1399 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 41 theme compound 693:700 arg1 solutions 702:710 pure compound solutions 688:710 pure compound solutions 688:710 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 4 42 theme matrix 863:868 arg1 fractogram 837:846 an AF4//MALS/dRI/UV fractogram 817:846 an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage 817:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 0 43 theme classes 40:46 arg1 Characterization 0:15 Characterization 0:15 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 0:152 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 7 44 theme AF4 1225:1227 arg1 fractogram 1229:1238 the AF4 fractogram 1221:1238 the AF4 fractogram of beer 1221:1246 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 0 45 theme cereal-based 51:62 arg1 beverages 64:72 cereal-based beverages 51:72 cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system 51:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 7 46 theme molar 1549:1553 arg1 mixture 1589:1595 a mixture 1587:1595 a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association 1587:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 46 theme molar 1549:1553 arg1 fraction 1560:1567 the high molar mass fraction 1540:1567 the high molar mass fraction 1540:1567 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 47 theme index 543:547 arg1 detection 549:557 refractive index detection 532:557 refractive index detection 532:557 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 8 48 theme complete 1832:1839 arg1 range 1852:1856 the complete molar mass range 1828:1856 the complete molar mass range 1828:1856 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 7 49 dep attributed 1441:1450 arg1 2 1403:1403 2 1403:1403 2 1403:1403 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 50 theme UV 568:569 arg1 UV 582:583 UV 582:583 UV 582:583 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 50 theme UV 568:569 arg1 detection 571:579 UV detection 568:579 UV detection (UV) 568:584 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 7 51 dep identified 1573:1582 arg1 3 1537:1537 3 1537:1537 3 1537:1537 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 5 52 theme polymer 1076:1082 arg1 standards 1084:1092 purified polymer standards 1067:1092 purified polymer standards 1067:1092 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 7 53 theme mass 1307:1310 arg1 fraction 1312:1319 the low molar mass fraction 1293:1319 the low molar mass fraction 1293:1319 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 54 theme AF4 1902:1904 arg1 fractogram 1906:1915 an AF4 fractogram 1899:1915 an AF4 fractogram 1899:1915 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 5 55 with alignments 1051:1060 arg1 standards 1084:1092 purified polymer standards 1067:1092 purified polymer standards 1067:1092 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 2 56 theme beverages 419:427 arg1 parameters 405:414 different technological and especially sensory parameters 358:414 different technological and especially sensory parameters of beverages 358:427 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 7 57 theme low 1297:1299 arg1 fraction 1312:1319 the low molar mass fraction 1293:1319 the low molar mass fraction 1293:1319 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 2 58 theme degradation 330:340 arg1 products 342:349 their degradation products 324:349 their degradation products 324:349 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 7 59 theme polysaccharides 1515:1529 arg1 coelution 1491:1499 a coelution 1489:1499 a coelution of non-starch polysaccharides 1489:1529 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 60 theme targeted 1939:1946 arg1 design 1948:1953 design 1948:1953 design 1948:1953 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 5 61 with precipitations 1009:1022 arg1 standards 1084:1092 purified polymer standards 1067:1092 purified polymer standards 1067:1092 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 1 62 contain contain 177:183 arg2 mixture 195:201 a complex mixture 185:201 a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols 185:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 62 contain contain 177:183 arg1 beverages 167:175 Cereal-based beverages 154:175 Cereal-based beverages 154:175 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 2 63 theme technological 368:380 arg1 parameters 405:414 different technological and especially sensory parameters 358:414 different technological and especially sensory parameters of beverages 358:427 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 1 64 theme polymeric 214:222 arg1 polyphenols 280:290 polyphenols 280:290 polyphenols 280:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 64 theme polymeric 214:222 arg1 polysaccharides 249:263 polysaccharides 249:263 polysaccharides 249:263 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 64 theme polymeric 214:222 arg1 macromolecules 224:237 various polymeric macromolecules 206:237 various polymeric macromolecules including polysaccharides, peptides, and polyphenols 206:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 1 64 theme polymeric 214:222 arg1 peptides 266:273 peptides 266:273 peptides 266:273 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 4 65 theme yeast-fermented 881:895 arg1 beverage 910:917 a yeast-fermented cereal-based beverage 879:917 a yeast-fermented cereal-based beverage 879:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 4 65 theme yeast-fermented 881:895 arg1 beer 873:876 beer 873:876 beer 873:876 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 7 66 theme protein-polyphenol 1455:1472 arg1 complexes 1474:1482 protein-polyphenol complexes 1455:1482 protein-polyphenol complexes with a coelution of non-starch polysaccharides 1455:1529 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 0 67 with fractionation 109:121 arg1 system 146:151 a multi-detection system 128:151 a multi-detection system 128:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 8 68 theme incomplete 1757:1766 arg1 hydrolysis 1775:1784 incomplete starch hydrolysis 1757:1784 incomplete starch hydrolysis 1757:1784 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 3 69 theme mass 625:628 arg1 analysis 643:650 molar mass distribution analysis 619:650 molar mass distribution analysis 619:650 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 9 70 from polymers 1973:1980 arg1 beverages 2008:2016 fermented cereal-based beverages 1985:2016 fermented cereal-based beverages 1985:2016 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 7 71 with mixture 1589:1595 arg1 content 1675:1681 a low content 1669:1681 a low content of polysaccharide-protein association 1669:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 5 72 theme substance 966:974 arg1 classes 976:982 polymer substance classes 958:982 polymer substance classes 958:982 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 7 73 theme molar 1417:1421 arg1 fraction 1428:1435 the middle molar mass fraction 1406:1435 the middle molar mass fraction 1406:1435 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 74 theme cereal-based 1995:2006 arg1 beverages 2008:2016 fermented cereal-based beverages 1985:2016 fermented cereal-based beverages 1985:2016 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 3 75 theme macromolecules 655:668 arg1 analysis 643:650 molar mass distribution analysis 619:650 molar mass distribution analysis 619:650 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 75 theme macromolecules 655:668 arg1 structure 605:613 structure 605:613 structure 605:613 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 1 76 theme Cereal-based 154:165 arg1 beverages 167:175 Cereal-based beverages 154:175 Cereal-based beverages 154:175 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 7 77 theme polysaccharides 1614:1628 arg1 mixture 1589:1595 a mixture 1587:1595 a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association 1587:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 77 theme polysaccharides 1614:1628 arg1 fraction 1560:1567 the high molar mass fraction 1540:1567 the high molar mass fraction 1540:1567 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 9 78 from design 1948:1953 arg1 beverages 2008:2016 fermented cereal-based beverages 1985:2016 fermented cereal-based beverages 1985:2016 The ability to assess the components of an AF4 fractogram is beneficial for the targeted design and evaluation of polymers in fermented cereal-based beverages and for controlling and monitoring quality parameters. 28735452 0 79 theme asymmetrical 80:91 arg1 fractionation 109:121 asymmetrical flow field-flow fractionation 80:121 asymmetrical flow field-flow fractionation with a multi-detection system 80:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 2 80 theme molar 297:301 arg1 mass 303:306 The molar mass 293:306 The molar mass of polymers and their degradation products 293:349 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 5 81 theme fractogram 934:943 arg1 fractions 945:953 fractogram fractions 934:953 fractogram fractions 934:953 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 7 82 theme cell 1604:1607 arg1 polysaccharides 1614:1628 the cell wall polysaccharides 1600:1628 the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) 1600:1662 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 0 83 theme field-flow 98:107 arg1 fractionation 109:121 asymmetrical flow field-flow fractionation 80:121 asymmetrical flow field-flow fractionation with a multi-detection system 80:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 1 84 theme complex 187:193 arg1 mixture 195:201 a complex mixture 185:201 a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols 185:290 Cereal-based beverages contain a complex mixture of various polymeric macromolecules including polysaccharides, peptides, and polyphenols. 28735452 8 85 attach derived 1744:1750 arg1 hydrolysis 1775:1784 incomplete starch hydrolysis 1757:1784 incomplete starch hydrolysis 1757:1784 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 8 85 attach derived 1744:1750 arg2 dextrins 1735:1742 dextrins 1735:1742 dextrins derived from incomplete starch hydrolysis 1735:1784 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 4 86 theme study 735:739 arg1 objective 717:725 The objective 713:725 The objective of this study 713:739 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 7 87 theme different 1268:1276 arg1 fractions 1278:1286 different fractions 1268:1286 different fractions 1268:1286 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 8 88 theme molar 1841:1845 arg1 range 1852:1856 the complete molar mass range 1828:1856 the complete molar mass range 1828:1856 In addition, dextrins derived from incomplete starch hydrolysis were identified in all fractions and over the complete molar mass range. 28735452 5 89 theme enzymatic 1025:1033 arg1 hydrolysis 1035:1044 enzymatic hydrolysis 1025:1044 enzymatic hydrolysis 1025:1044 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 3 90 theme Asymmetrical 430:441 arg1 fractionation 459:471 Asymmetrical flow field-flow fractionation 430:471 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) 430:584 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 90 theme Asymmetrical 430:441 arg1 technique 591:599 a technique 589:599 a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions 589:710 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 90 theme Asymmetrical 430:441 arg1 AF4 474:476 AF4 474:476 AF4 474:476 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 4 91 theme polymer 798:804 arg1 classes 806:812 the polymer classes 794:812 the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage 794:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 0 92 theme multi-detection 130:144 arg1 system 146:151 a multi-detection system 128:151 a multi-detection system 128:151 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 7 93 dep assigned 1325:1332 arg1 1 1290:1290 1 1290:1290 1 1290:1290 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 94 theme glycosylation 1387:1399 arg1 degrees 1376:1382 different degrees 1366:1382 different degrees of glycosylation 1366:1399 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 95 theme beer 1243:1246 arg1 fractogram 1229:1238 the AF4 fractogram 1221:1238 the AF4 fractogram of beer 1221:1246 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 96 theme light 504:508 arg1 scattering 510:519 multi-angle light scattering 492:519 multi-angle light scattering (MALS) 492:526 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 96 theme light 504:508 arg1 dRI 560:562 dRI 560:562 dRI 560:562 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 96 theme light 504:508 arg1 MALS 522:525 MALS 522:525 MALS 522:525 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 0 97 theme substance 30:38 arg1 classes 40:46 polymeric substance classes 20:46 polymeric substance classes 20:46 Characterization of polymeric substance classes in cereal-based beverages using asymmetrical flow field-flow fractionation with a multi-detection system. 28735452 7 98 theme polysaccharide-protein 1686:1707 arg1 association 1709:1719 polysaccharide-protein association 1686:1719 polysaccharide-protein association 1686:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 3 99 theme field-flow 448:457 arg1 fractionation 459:471 Asymmetrical flow field-flow fractionation 430:471 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) 430:584 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 99 theme field-flow 448:457 arg1 technique 591:599 a technique 589:599 a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions 589:710 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 99 theme field-flow 448:457 arg1 AF4 474:476 AF4 474:476 AF4 474:476 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 100 theme pure 688:691 arg1 solutions 702:710 pure compound solutions 688:710 pure compound solutions 688:710 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 3 101 theme refractive 532:541 arg1 detection 549:557 refractive index detection 532:557 refractive index detection 532:557 Asymmetrical flow field-flow fractionation (AF4) coupled with multi-angle light scattering (MALS) and refractive index detection (dRI) or UV detection (UV) is a technique for structure and molar mass distribution analysis of macromolecules commonly used for pure compound solutions. 28735452 7 102 with complexes 1474:1482 arg1 coelution 1491:1499 a coelution 1489:1499 a coelution of non-starch polysaccharides 1489:1529 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 103 theme proteinaceous 1337:1349 arg1 molecules 1351:1359 proteinaceous molecules 1337:1359 proteinaceous molecules with different degrees of glycosylation 1337:1399 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 4 104 theme complex 855:861 arg1 matrix 863:868 the complex matrix 851:868 the complex matrix 851:868 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 2 105 theme sensory 397:403 arg1 parameters 405:414 different technological and especially sensory parameters 358:414 different technological and especially sensory parameters of beverages 358:427 The molar mass of polymers and their degradation products affect different technological and especially sensory parameters of beverages. 28735452 7 106 theme mass 1555:1558 arg1 mixture 1589:1595 a mixture 1587:1595 a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association 1587:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 106 theme mass 1555:1558 arg1 fraction 1560:1567 the high molar mass fraction 1540:1567 the high molar mass fraction 1540:1567 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 4 107 theme AF4//MALS/dRI/UV 820:835 arg1 fractogram 837:846 an AF4//MALS/dRI/UV fractogram 817:846 an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage 817:917 The objective of this study was to develop a systematic approach for identifying the polymer classes in an AF4//MALS/dRI/UV fractogram of the complex matrix in beer, a yeast-fermented cereal-based beverage. 28735452 7 108 theme high 1544:1547 arg1 mixture 1589:1595 a mixture 1587:1595 a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association 1587:1719 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 7 108 theme high 1544:1547 arg1 fraction 1560:1567 the high molar mass fraction 1540:1567 the high molar mass fraction 1540:1567 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 28735452 5 109 theme purified 1067:1074 arg1 standards 1084:1092 purified polymer standards 1067:1092 purified polymer standards 1067:1092 Assignment of fractogram fractions to polymer substance classes was achieved by targeted precipitations, enzymatic hydrolysis, and alignments with purified polymer standards. 28735452 7 110 theme molar 1301:1305 arg1 fraction 1312:1319 the low molar mass fraction 1293:1319 the low molar mass fraction 1293:1319 Using these techniques, the AF4 fractogram of beer was classified into different fractions: (1) the low molar mass fraction was assigned to proteinaceous molecules with different degrees of glycosylation, (2) the middle molar mass fraction was attributed to protein-polyphenol complexes with a coelution of non-starch polysaccharides, and (3) the high molar mass fraction was identified as a mixture of the cell wall polysaccharides (i.e., β-glucan and arabinoxylan) with a low content of polysaccharide-protein association. 25124036 9 0 theme gastric 1679:1685 arg1 epithelia 1687:1695 gastric epithelia 1679:1695 gastric epithelia 1679:1695 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 0 1 theme α-GlcNAc-capped 46:60 arg1 glycans 68:74 α-GlcNAc-capped mucin glycans 46:74 α-GlcNAc-capped mucin glycans 46:74 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 7 2 theme TFF2-mucin 1395:1404 arg1 binding 1406:1412 TFF2-mucin binding 1395:1412 TFF2-mucin binding 1395:1412 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 3 3 theme deep 557:560 arg1 glands 570:575 deep gastric glands 557:575 deep gastric glands 557:575 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 1 4 theme gastric 182:188 arg1 cancer 190:195 gastric cancer 182:195 gastric cancer 182:195 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 9 5 theme functional 1703:1712 arg1 relationship 1714:1725 a functional relationship 1701:1725 a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans 1701:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 6 6 theme porcine 1140:1146 arg1 mucin 1156:1160 porcine stomach mucin 1140:1160 porcine stomach mucin 1140:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 8 7 from binding 1518:1524 arg1 GalNAc-ol 1602:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol 1559:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1) 1559:1623 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 8 7 from binding 1518:1524 arg1 Structure 1612:1620 Structure 1 1612:1622 Structure 1 1612:1622 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 0 8 theme Helicobacter 109:120 arg1 pylori 122:127 Helicobacter pylori 109:127 Helicobacter pylori 109:127 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 5 9 theme group 721:725 arg1 O-glycans 734:742 blood group active O-glycans 715:742 blood group active O-glycans of the Lewis-type 715:760 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 5 10 theme epithelial 833:842 arg1 cells 844:848 epithelial cells 833:848 epithelial cells 833:848 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 10 11 theme general 1877:1883 arg1 aspects 1896:1902 more general functional aspects 1872:1902 more general functional aspects of TFF members 1872:1917 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 2 12 theme trefoil 249:255 arg1 peptides 264:271 The trefoil factor peptides 245:271 The trefoil factor peptides 245:271 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 2 12 theme trefoil 249:255 arg1 family 279:284 a family 277:284 a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract 277:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 5 13 theme H. 923:924 arg1 antibiotic 953:962 antibiotic 953:962 antibiotic 953:962 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 5 13 theme H. 923:924 arg1 growth 933:938 H. pylori growth 923:938 H. pylori growth 923:938 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 8 14 theme isomers 1480:1486 arg1 branches 1460:1467 the C3/C6-linked branches 1443:1467 the C3/C6-linked branches of the two isomers 1443:1486 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 7 15 theme resonance 1326:1334 arg1 spectroscopy 1336:1347 (1)H nuclear magnetic resonance spectroscopy 1304:1347 (1)H nuclear magnetic resonance spectroscopy 1304:1347 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 4 16 theme H. 685:686 arg1 infection 695:703 H. pylori infection 685:703 H. pylori infection 685:703 Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. 25124036 7 17 theme active 1236:1241 arg1 glycotope 1243:1251 the binding active glycotope 1224:1251 the binding active glycotope 1224:1251 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 5 18 theme gastric 883:889 arg1 mucins 891:896 gastric mucins 883:896 gastric mucins 883:896 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 6 19 theme gastric 987:993 arg1 glycoform 995:1003 the gastric glycoform 983:1003 the gastric glycoform of TFF2 983:1011 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 6 19 theme gastric 987:993 arg1 lectin 1038:1043 a calcium-independent lectin 1016:1043 a calcium-independent lectin 1016:1043 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 9 20 theme α1,4-GlcNAc-capped 1779:1796 arg1 glycans 1804:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 0 21 theme mucin 62:66 arg1 glycans 68:74 α-GlcNAc-capped mucin glycans 46:74 α-GlcNAc-capped mucin glycans 46:74 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 8 22 theme TFF2 1513:1516 arg1 binding 1518:1524 highly specific TFF2 binding 1497:1524 highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1) 1497:1623 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 7 23 theme isomers 1212:1218 arg1 glycotope 1243:1251 the binding active glycotope 1224:1251 the binding active glycotope 1224:1251 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 23 theme isomers 1212:1218 arg1 assignments 1178:1188 The structural assignments 1163:1188 The structural assignments of two hexasaccharide isomers 1163:1218 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 1 24 theme Helicobacter 130:141 arg1 infection 150:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 1 24 theme Helicobacter 130:141 arg1 cause 173:177 the major cause 163:177 the major cause of gastric cancer 163:195 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 2 25 theme epithelial 384:393 arg1 repair 395:400 epithelial repair 384:400 epithelial repair 384:400 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 9 26 dep H. 1746:1747 arg1 pylori 1749:1754 H. pylori 1746:1754 H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans 1746:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 7 27 theme magnetic 1317:1324 arg1 resonance 1326:1334 (1)H nuclear magnetic resonance 1304:1334 (1)H nuclear magnetic resonance spectroscopy 1304:1347 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 8 28 theme 6-linked 1533:1540 arg1 trisaccharide 1542:1554 the 6-linked trisaccharide 1529:1554 the 6-linked trisaccharide 1529:1554 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 7 29 theme binding 1228:1234 arg1 glycotope 1243:1251 the binding active glycotope 1224:1251 the binding active glycotope 1224:1251 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 0 30 theme Human 0:4 arg1 factor 14:19 Human trefoil factor 2 0:21 Human trefoil factor 2 0:21 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 0 30 theme Human 0:4 arg1 lectin 28:33 a lectin 26:33 a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori 26:127 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 10 31 theme Lectin-carbohydrate 1813:1831 arg1 interaction 1833:1843 Lectin-carbohydrate interaction 1813:1843 Lectin-carbohydrate interaction 1813:1843 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 5 32 theme active 727:732 arg1 O-glycans 734:742 blood group active O-glycans 715:742 blood group active O-glycans of the Lewis-type 715:760 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 7 33 theme H 1307:1307 arg1 resonance 1326:1334 (1)H nuclear magnetic resonance 1304:1334 (1)H nuclear magnetic resonance spectroscopy 1304:1347 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 2 34 theme gastrointestinal 413:428 arg1 tract 430:434 the gastrointestinal tract 409:434 the gastrointestinal tract 409:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 5 35 theme mucin 814:818 arg1 layer 820:824 the surface mucin layer 802:824 the surface mucin layer 802:824 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 6 36 theme calcium-independent 1018:1036 arg1 glycoform 995:1003 the gastric glycoform 983:1003 the gastric glycoform of TFF2 983:1011 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 6 36 theme calcium-independent 1018:1036 arg1 lectin 1038:1043 a calcium-independent lectin 1016:1043 a calcium-independent lectin 1016:1043 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 5 37 theme H. 780:781 arg1 adhesion 790:797 H. pylori adhesion 780:797 H. pylori adhesion to the surface mucin layer and to epithelial cells 780:848 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 7 38 dep H 1307:1307 arg1 1 1305:1305 1 1305:1305 1 1305:1305 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 10 39 theme functional 1885:1894 arg1 aspects 1896:1902 more general functional aspects 1872:1902 more general functional aspects of TFF members 1872:1917 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 3 40 theme surface 486:492 arg1 epithelium 494:503 the gastric surface epithelium 474:503 the gastric surface epithelium 474:503 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 5 41 theme adhesion 790:797 arg1 basis 771:775 the basis 767:775 the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells 767:848 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 7 42 theme linkage 1286:1292 arg1 analysis 1294:1301 linkage analysis 1286:1301 linkage analysis 1286:1301 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 1 43 theme important 212:220 arg1 challenge 234:242 an important health care challenge 209:242 an important health care challenge 209:242 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 9 44 theme lectin 1638:1643 arg1 TFF2 1645:1648 lectin TFF2 1638:1648 lectin TFF2 1638:1648 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 6 45 theme α1,4-GlcNAc-capped 1092:1109 arg1 hexasaccharides 1111:1125 O-linked α1,4-GlcNAc-capped hexasaccharides 1083:1125 O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin 1083:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 10 46 theme members 1911:1917 arg1 aspects 1896:1902 more general functional aspects 1872:1902 more general functional aspects of TFF members 1872:1917 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 1 47 theme care 229:232 arg1 challenge 234:242 an important health care challenge 209:242 an important health care challenge 209:242 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 8 48 link 6-linked 1533:1540 arg1 trisaccharide 1542:1554 the 6-linked trisaccharide 1529:1554 the 6-linked trisaccharide 1529:1554 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 10 49 theme cell 1949:1952 arg1 receptors 1964:1972 cell signaling receptors 1949:1972 cell signaling receptors 1949:1972 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 6 50 gly glycoform 995:1003 arg1 TFF2 1008:1011 TFF2 1008:1011 TFF2 1008:1011 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 6 51 theme high 1063:1066 arg1 specificity 1068:1078 high specificity 1063:1078 high specificity 1063:1078 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 1 52 theme major 167:171 arg1 infection 150:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 1 52 theme major 167:171 arg1 cause 173:177 the major cause 163:177 the major cause of gastric cancer 163:195 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 3 53 with concert 536:542 arg1 TFF2 549:552 TFF2 549:552 TFF2 549:552 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 9 54 theme epithelia 1687:1695 arg1 protection 1665:1674 protection 1665:1674 protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans 1665:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 2 55 theme conserved 302:310 arg1 proteins 312:319 small highly conserved proteins 289:319 small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract 289:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 5 56 theme α1,4-GlcNAc-capped 851:868 arg1 O-glycans 870:878 α1,4-GlcNAc-capped O-glycans 851:878 α1,4-GlcNAc-capped O-glycans on gastric mucins 851:896 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 5 57 dep H. 780:781 arg1 pylori 783:788 H. pylori 780:788 H. pylori adhesion to the surface mucin layer and to epithelial cells 780:848 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 6 58 theme stomach 1148:1154 arg1 mucin 1156:1160 porcine stomach mucin 1140:1160 porcine stomach mucin 1140:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 4 59 theme increased 659:667 arg1 expression 669:678 increased expression 659:678 increased expression upon H. pylori infection 659:703 Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. 25124036 2 60 theme small 289:293 arg1 proteins 312:319 small highly conserved proteins 289:319 small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract 289:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 1 61 theme cancer 190:195 arg1 infection 150:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 1 61 theme cancer 190:195 arg1 cause 173:177 the major cause 163:177 the major cause of gastric cancer 163:195 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 7 62 theme binding 1406:1412 arg1 spectrometry 1272:1283 mass spectrometry 1267:1283 mass spectrometry 1267:1283 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 62 theme binding 1406:1412 arg1 competition 1380:1390 lectin competition 1373:1390 lectin competition 1373:1390 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 62 theme binding 1406:1412 arg1 analysis 1294:1301 linkage analysis 1286:1301 linkage analysis 1286:1301 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 62 theme binding 1406:1412 arg1 spectroscopy 1336:1347 (1)H nuclear magnetic resonance spectroscopy 1304:1347 (1)H nuclear magnetic resonance spectroscopy 1304:1347 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 62 theme binding 1406:1412 arg1 inhibition 1357:1366 glycan inhibition 1350:1366 glycan inhibition 1350:1366 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 8 63 theme C3/C6-linked 1447:1458 arg1 branches 1460:1467 the C3/C6-linked branches 1443:1467 the C3/C6-linked branches of the two isomers 1443:1486 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 4 64 theme gland 609:613 arg1 components 583:592 Both components 578:592 Both components of the gastric gland 578:613 Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. 25124036 5 65 theme blood 715:719 arg1 O-glycans 734:742 blood group active O-glycans 715:742 blood group active O-glycans of the Lewis-type 715:760 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 7 66 theme lectin 1373:1378 arg1 competition 1380:1390 lectin competition 1373:1390 lectin competition 1373:1390 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 7 67 theme hexasaccharide 1197:1210 arg1 isomers 1212:1218 two hexasaccharide isomers 1193:1218 two hexasaccharide isomers 1193:1218 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 3 68 theme gastric 562:568 arg1 glands 570:575 deep gastric glands 557:575 deep gastric glands 557:575 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 2 69 theme factor 257:262 arg1 peptides 264:271 The trefoil factor peptides 245:271 The trefoil factor peptides 245:271 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 2 69 theme factor 257:262 arg1 family 279:284 a family 277:284 a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract 277:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 6 70 theme TFF2 1008:1011 arg1 glycoform 995:1003 the gastric glycoform 983:1003 the gastric glycoform of TFF2 983:1011 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 6 70 theme TFF2 1008:1011 arg1 lectin 1038:1043 a calcium-independent lectin 1016:1043 a calcium-independent lectin 1016:1043 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 7 71 theme glycan 1350:1355 arg1 inhibition 1357:1366 glycan inhibition 1350:1366 glycan inhibition 1350:1366 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 5 72 theme Lewis-type 751:760 arg1 O-glycans 734:742 blood group active O-glycans 715:742 blood group active O-glycans of the Lewis-type 715:760 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 5 73 from O-glycans 870:878 arg1 mucins 891:896 gastric mucins 883:896 gastric mucins 883:896 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 1 74 theme pylori 143:148 arg1 infection 150:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection 130:158 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 1 74 theme pylori 143:148 arg1 cause 173:177 the major cause 163:177 the major cause of gastric cancer 163:195 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 9 75 theme antibiotic 1768:1777 arg1 glycans 1804:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 8 76 theme specific 1504:1511 arg1 binding 1518:1524 highly specific TFF2 binding 1497:1524 highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1) 1497:1623 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 9 77 theme mucin 1798:1802 arg1 glycans 1804:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 antibiotic α1,4-GlcNAc-capped mucin glycans 1768:1810 Supposedly, lectin TFF2 is involved in protection of gastric epithelia via a functional relationship to defense against H. pylori launched by antibiotic α1,4-GlcNAc-capped mucin glycans. 25124036 4 78 theme pylori 688:693 arg1 infection 695:703 H. pylori infection 685:703 H. pylori infection 685:703 Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. 25124036 0 79 theme antibiotic 81:90 arg1 activity 92:99 antibiotic activity 81:99 antibiotic activity against Helicobacter pylori 81:127 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 5 80 dep H. 923:924 arg1 pylori 926:931 H. pylori 923:931 H. pylori growth 923:938 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 3 81 theme gastric 478:484 arg1 epithelium 494:503 the gastric surface epithelium 474:503 the gastric surface epithelium 474:503 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 7 82 theme nuclear 1309:1315 arg1 resonance 1326:1334 (1)H nuclear magnetic resonance 1304:1334 (1)H nuclear magnetic resonance spectroscopy 1304:1347 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 0 83 theme trefoil 6:12 arg1 factor 14:19 Human trefoil factor 2 0:21 Human trefoil factor 2 0:21 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 0 83 theme trefoil 6:12 arg1 lectin 28:33 a lectin 26:33 a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori 26:127 Human trefoil factor 2 is a lectin that binds α-GlcNAc-capped mucin glycans with antibiotic activity against Helicobacter pylori. 25124036 8 84 link C3/C6-linked 1447:1458 arg1 branches 1460:1467 the C3/C6-linked branches 1443:1467 the C3/C6-linked branches of the two isomers 1443:1486 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 10 85 contain have 1849:1852 arg2 impact 1862:1867 an impact 1859:1867 an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors 1859:1972 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 10 85 contain have 1849:1852 arg1 interaction 1833:1843 Lectin-carbohydrate interaction 1813:1843 Lectin-carbohydrate interaction 1813:1843 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 8 86 theme GlcNAcα1-4Galβ1-4GlcNAcβ1-6 1559:1585 arg1 GalNAc-ol 1602:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol 1559:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1) 1559:1623 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 8 86 theme GlcNAcα1-4Galβ1-4GlcNAcβ1-6 1559:1585 arg1 Structure 1612:1620 Structure 1 1612:1622 Structure 1 1612:1622 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 5 87 theme surface 806:812 arg1 layer 820:824 the surface mucin layer 802:824 the surface mucin layer 802:824 Although blood group active O-glycans of the Lewis-type form the basis of H. pylori adhesion to the surface mucin layer and to epithelial cells, α1,4-GlcNAc-capped O-glycans on gastric mucins were proposed to inhibit H. pylori growth as a natural antibiotic. 25124036 10 88 from impact 1862:1867 arg1 aspects 1896:1902 more general functional aspects 1872:1902 more general functional aspects of TFF members 1872:1917 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 8 89 theme Fucα1-2Galβ1-3 1587:1600 arg1 GalNAc-ol 1602:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol 1559:1610 GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1) 1559:1623 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 8 89 theme Fucα1-2Galβ1-3 1587:1600 arg1 Structure 1612:1620 Structure 1 1612:1622 Structure 1 1612:1622 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 10 90 theme TFF 1907:1909 arg1 members 1911:1917 TFF members 1907:1917 TFF members 1907:1917 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 6 91 from hexasaccharides 1111:1125 arg1 mucin 1156:1160 porcine stomach mucin 1140:1160 porcine stomach mucin 1140:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 6 91 from hexasaccharides 1111:1125 arg1 human 1130:1134 human 1130:1134 human 1130:1134 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 8 92 attach derived 1430:1436 arg1 branches 1460:1467 the C3/C6-linked branches 1443:1467 the C3/C6-linked branches of the two isomers 1443:1486 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 8 92 attach derived 1430:1436 arg2 Neoglycolipids 1415:1428 Neoglycolipids 1415:1428 Neoglycolipids derived from the C3/C6-linked branches of the two isomers 1415:1486 Neoglycolipids derived from the C3/C6-linked branches of the two isomers revealed highly specific TFF2 binding to the 6-linked trisaccharide in GlcNAcα1-4Galβ1-4GlcNAcβ1-6(Fucα1-2Galβ1-3)GalNAc-ol(Structure 1). 25124036 3 93 dep H. 437:438 arg1 pylori 440:445 H. pylori 437:445 H. pylori 437:445 H. pylori colocalizes with MUC5AC at the gastric surface epithelium, but not with MUC6 secreted in concert with TFF2 by deep gastric glands. 25124036 2 94 theme essential 346:354 arg1 roles 356:360 essential roles 346:360 essential roles 346:360 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 7 95 theme mass 1267:1270 arg1 spectrometry 1272:1283 mass spectrometry 1267:1283 mass spectrometry 1267:1283 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 25124036 6 96 link O-linked 1083:1090 arg1 hexasaccharides 1111:1125 O-linked α1,4-GlcNAc-capped hexasaccharides 1083:1125 O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin 1083:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 2 97 theme proteins 312:319 arg1 peptides 264:271 The trefoil factor peptides 245:271 The trefoil factor peptides 245:271 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 2 97 theme proteins 312:319 arg1 family 279:284 a family 277:284 a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract 277:434 The trefoil factor peptides are a family of small highly conserved proteins that are claimed to play essential roles in cytoprotection and epithelial repair within the gastrointestinal tract. 25124036 1 98 theme health 222:227 arg1 challenge 234:242 an important health care challenge 209:242 an important health care challenge 209:242 Helicobacter pylori infection is the major cause of gastric cancer and remains an important health care challenge. 25124036 6 99 theme O-linked 1083:1090 arg1 hexasaccharides 1111:1125 O-linked α1,4-GlcNAc-capped hexasaccharides 1083:1125 O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin 1083:1160 We show here that the gastric glycoform of TFF2 is a calcium-independent lectin, which binds with high specificity to O-linked α1,4-GlcNAc-capped hexasaccharides on human and porcine stomach mucin. 25124036 4 100 theme gastric 601:607 arg1 gland 609:613 the gastric gland 597:613 the gastric gland 597:613 Both components of the gastric gland secretome associate non-covalently and show increased expression upon H. pylori infection. 25124036 10 101 theme signaling 1954:1962 arg1 receptors 1964:1972 cell signaling receptors 1949:1972 cell signaling receptors 1949:1972 Lectin-carbohydrate interaction may have also an impact on more general functional aspects of TFF members by mediating their binding to cell signaling receptors. 25124036 7 102 theme structural 1167:1176 arg1 assignments 1178:1188 The structural assignments 1163:1188 The structural assignments of two hexasaccharide isomers 1163:1218 The structural assignments of two hexasaccharide isomers and the binding active glycotope were based on mass spectrometry, linkage analysis, (1)H nuclear magnetic resonance spectroscopy, glycan inhibition, and lectin competition of TFF2-mucin binding. 26918373 6 0 from variations 924:933 arg1 acid 1007:1010 sialic acid 1000:1010 sialic acid 1000:1010 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 0 from variations 924:933 arg1 structures 964:973 the terminal carbohydrate structures 938:973 the terminal carbohydrate structures such as galactose versus sialic acid epitopes 938:1019 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 0 from variations 924:933 arg1 galactose 983:991 galactose 983:991 galactose 983:991 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 1 1 theme product 165:171 arg1 efficacy 193:200 efficacy 193:200 efficacy 193:200 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 1 1 theme product 165:171 arg1 safety 182:187 safety 182:187 safety 182:187 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 1 1 theme product 165:171 arg1 quality 173:179 product quality 165:179 product quality 165:179 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 0 2 theme proteins 72:79 arg1 glycosylation 43:55 glycosylation 43:55 glycosylation of therapeutic proteins 43:79 The use of lectin microarray for assessing glycosylation of therapeutic proteins. 26918373 2 3 theme analytical 358:367 arg1 challenge 369:377 a daunting analytical challenge 347:377 a daunting analytical challenge 347:377 However, the complexity of protein glycosylation poses a daunting analytical challenge. 26918373 7 4 theme glycoproteins 1123:1135 arg1 patterns 1099:1106 glycan patterns 1092:1106 glycan patterns of therapeutic glycoproteins 1092:1135 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 26918373 4 5 theme proteins 648:655 arg1 panel 624:628 a panel 622:628 a panel of 15 therapeutic proteins, including 8 monoclonal antibodies 622:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 1 6 theme product 271:277 arg1 cycles 284:289 product life cycles 271:289 product life cycles 271:289 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 4 7 theme commercial 480:489 arg1 chips 498:502 commercial lectin chips 480:502 commercial lectin chips 480:502 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 1 8 theme life 279:282 arg1 cycles 284:289 product life cycles 271:289 product life cycles 271:289 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 5 9 with consistent 778:787 arg1 patterns 811:818 the known glycan patterns 794:818 the known glycan patterns for these glycoproteins 794:842 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 3 10 theme microarray 432:441 arg1 utility 412:418 the utility 408:418 the utility of a lectin microarray for assessing protein glycans 408:471 In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. 26918373 5 11 theme glycan 742:747 arg1 profiles 749:756 glycan profiles 742:756 glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins 742:842 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 5 12 gly glycoproteins 830:842 arg1 glycoproteins 830:842 these glycoproteins 824:842 these glycoproteins 824:842 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 1 13 attach attached 107:114 arg1 glycoproteins 131:143 therapeutic glycoproteins 119:143 therapeutic glycoproteins 119:143 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 1 13 attach attached 107:114 arg2 Glycans 82:88 Glycans 82:88 Glycans 82:88 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 5 14 theme glycan 804:809 arg1 patterns 811:818 the known glycan patterns 794:818 the known glycan patterns for these glycoproteins 794:842 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 4 15 theme lectin 491:496 arg1 chips 498:502 commercial lectin chips 480:502 commercial lectin chips 480:502 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 7 16 theme glycan 1092:1097 arg1 patterns 1099:1106 glycan patterns 1092:1106 glycan patterns of therapeutic glycoproteins 1092:1135 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 26918373 2 17 theme protein 319:325 arg1 glycosylation 327:339 protein glycosylation 319:339 protein glycosylation 319:339 However, the complexity of protein glycosylation poses a daunting analytical challenge. 26918373 3 18 theme lectin 425:430 arg1 microarray 432:441 a lectin microarray 423:441 a lectin microarray 423:441 In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. 26918373 0 19 theme lectin 11:16 arg1 microarray 18:27 lectin microarray 11:27 lectin microarray 11:27 The use of lectin microarray for assessing glycosylation of therapeutic proteins. 26918373 5 20 theme Lectin 693:698 arg1 signals 708:714 Lectin binding signals 693:714 Lectin binding signals 693:714 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 4 21 theme panel 624:628 arg1 patterns 610:617 the lectin binding patterns 591:617 the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies 591:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 5 22 theme binding 700:706 arg1 signals 708:714 Lectin binding signals 693:714 Lectin binding signals 693:714 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 4 23 theme monoclonal 670:679 arg1 antibodies 681:690 8 monoclonal antibodies 668:690 8 monoclonal antibodies 668:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 6 24 theme lectin-based 864:875 arg1 sensitive 911:919 sensitive 911:919 sensitive 911:919 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 24 theme lectin-based 864:875 arg1 microarray 877:886 the lectin-based microarray 860:886 the lectin-based microarray 860:886 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 25 located found 892:896 arg2 microarray 877:886 the lectin-based microarray 860:886 the lectin-based microarray 860:886 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 25 located found 892:896 arg2 sensitive 911:919 sensitive 911:919 sensitive 911:919 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 25 located found 892:896 arg1 particular 848:857 particular 848:857 particular 848:857 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 0 26 theme microarray 18:27 arg1 use 4:6 The use 0:6 The use of lectin microarray for assessing glycosylation of therapeutic proteins 0:79 The use of lectin microarray for assessing glycosylation of therapeutic proteins. 26918373 6 27 theme carbohydrate 951:962 arg1 acid 1007:1010 sialic acid 1000:1010 sialic acid 1000:1010 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 27 theme carbohydrate 951:962 arg1 structures 964:973 the terminal carbohydrate structures 938:973 the terminal carbohydrate structures such as galactose versus sialic acid epitopes 938:1019 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 27 theme carbohydrate 951:962 arg1 galactose 983:991 galactose 983:991 galactose 983:991 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 1 28 gly glycoproteins 131:143 arg1 glycoproteins 131:143 therapeutic glycoproteins 119:143 therapeutic glycoproteins 119:143 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 3 29 theme protein 457:463 arg1 glycans 465:471 protein glycans 457:471 protein glycans 457:471 In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. 26918373 6 30 theme terminal 942:949 arg1 acid 1007:1010 sialic acid 1000:1010 sialic acid 1000:1010 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 30 theme terminal 942:949 arg1 structures 964:973 the terminal carbohydrate structures 938:973 the terminal carbohydrate structures such as galactose versus sialic acid epitopes 938:1019 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 6 30 theme terminal 942:949 arg1 galactose 983:991 galactose 983:991 galactose 983:991 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 2 31 theme daunting 349:356 arg1 challenge 369:377 a daunting analytical challenge 347:377 a daunting analytical challenge 347:377 However, the complexity of protein glycosylation poses a daunting analytical challenge. 26918373 5 32 theme known 798:802 arg1 patterns 811:818 the known glycan patterns 794:818 the known glycan patterns for these glycoproteins 794:842 Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. 26918373 4 33 theme glycan 546:551 arg1 structures 553:562 distinct glycan structures 537:562 distinct glycan structures 537:562 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 7 34 theme therapeutic 1111:1121 arg1 glycoproteins 1123:1135 therapeutic glycoproteins 1111:1135 therapeutic glycoproteins 1111:1135 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 26918373 1 35 theme therapeutic 119:129 arg1 glycoproteins 131:143 therapeutic glycoproteins 119:143 therapeutic glycoproteins 119:143 Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. 26918373 4 36 theme distinct 537:544 arg1 structures 553:562 distinct glycan structures 537:562 distinct glycan structures 537:562 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 4 37 theme binding 602:608 arg1 patterns 610:617 the lectin binding patterns 591:617 the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies 591:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 7 38 theme lectin 1046:1051 arg1 microarray 1053:1062 lectin microarray 1046:1062 lectin microarray 1046:1062 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 26918373 6 39 dep galactose 983:991 arg1 epitopes 1012:1019 epitopes 1012:1019 epitopes 1012:1019 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 2 40 theme glycosylation 327:339 arg1 complexity 305:314 the complexity 301:314 the complexity of protein glycosylation 301:339 However, the complexity of protein glycosylation poses a daunting analytical challenge. 26918373 4 41 theme lectin 595:600 arg1 patterns 610:617 the lectin binding patterns 591:617 the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies 591:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 0 42 theme therapeutic 60:70 arg1 proteins 72:79 therapeutic proteins 60:79 therapeutic proteins 60:79 The use of lectin microarray for assessing glycosylation of therapeutic proteins. 26918373 7 43 gly glycoproteins 1123:1135 arg1 glycoproteins 1123:1135 therapeutic glycoproteins 1111:1135 therapeutic glycoproteins 1111:1135 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 26918373 4 44 contain contain 511:517 arg2 lectins 522:528 45 lectins 519:528 45 lectins toward distinct glycan structures 519:562 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 4 44 contain contain 511:517 arg1 chips 498:502 commercial lectin chips 480:502 commercial lectin chips 480:502 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 4 45 theme therapeutic 636:646 arg1 antibodies 681:690 8 monoclonal antibodies 668:690 8 monoclonal antibodies 668:690 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 4 45 theme therapeutic 636:646 arg1 proteins 648:655 15 therapeutic proteins 633:655 15 therapeutic proteins 633:655 Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. 26918373 6 46 theme sialic 1000:1005 arg1 acid 1007:1010 sialic acid 1000:1010 sialic acid 1000:1010 In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. 26918373 0 47 gly glycosylation 43:55 arg1 proteins 72:79 therapeutic proteins 60:79 therapeutic proteins 60:79 The use of lectin microarray for assessing glycosylation of therapeutic proteins. 26918373 7 48 used used 1073:1076 arg2 microarray 1053:1062 lectin microarray 1046:1062 lectin microarray 1046:1062 These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. 25186198 3 0 theme Protein 311:317 arg1 chaperone 465:473 a chaperone 463:473 a chaperone involved in the proper localization of the Notch receptor in certain contexts 463:551 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 0 theme Protein 311:317 arg1 enzyme 382:387 a soluble, ER localized enzyme 358:387 a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats 358:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 0 theme Protein 311:317 arg1 O-fucosyltransferase 319:338 Protein O-fucosyltransferase 1 311:340 Protein O-fucosyltransferase 1 (Ofut1/Pofut1) 311:355 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 0 theme Protein 311:317 arg1 Ofut1/Pofut1 343:354 Ofut1/Pofut1 343:354 Ofut1/Pofut1 343:354 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 1 dep soluble 360:366 arg1 localized 372:380 localized 372:380 localized 372:380 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 7 2 theme consensus 955:963 arg1 sequences 965:973 consensus sequences 955:973 consensus sequences for O-fucosylation 955:992 Though consensus sequences for O-fucosylation have been established and structures of both Pofuts have been studied, the mechanism of how they participate in protein folding is not known. 25186198 1 3 theme endoplasmic 136:146 arg1 ER 159:160 ER 159:160 ER 159:160 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 1 3 theme endoplasmic 136:146 arg1 reticulum 148:156 the endoplasmic reticulum 132:156 the endoplasmic reticulum (ER) 132:161 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 8 4 theme recent 1168:1173 arg1 advances 1175:1182 past and recent advances 1159:1182 past and recent advances made in novel roles for these protein O-glycosyltransferases 1159:1243 This article discusses past and recent advances made in novel roles for these protein O-glycosyltransferases. 25186198 0 5 from roles 6:10 arg1 folding 44:50 protein folding 36:50 protein folding 36:50 Novel roles for O-linked glycans in protein folding. 25186198 3 6 gly fucosylates 394:404 arg1 repeats 441:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 0 7 link O-linked 16:23 arg1 glycans 25:31 O-linked glycans 16:31 O-linked glycans 16:31 Novel roles for O-linked glycans in protein folding. 25186198 2 8 theme important 282:290 arg1 function 292:299 this important function 277:299 this important function 277:299 Recent work has shown that O-linked glycosylation and the corresponding glycosyltransferases also participate in this important function. 25186198 3 9 theme certain 536:542 arg1 contexts 544:551 certain contexts 536:551 certain contexts 536:551 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 4 10 theme related 564:570 arg1 Pofut2 554:559 Pofut2 554:559 Pofut2 554:559 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 4 10 theme related 564:570 arg1 enzyme 572:577 a related enzyme 562:577 a related enzyme that modifies Thrombospondin type I repeats (TSRs) 562:628 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 3 11 theme receptor 524:531 arg1 localization 498:509 the proper localization 487:509 the proper localization of the Notch receptor in certain contexts 487:551 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 12 from localization 498:509 arg1 contexts 544:551 certain contexts 536:551 certain contexts 536:551 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 4 13 theme proteins 726:733 arg1 control 700:706 the folding and quality control 676:706 the folding and quality control of TSR-containing proteins 676:733 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 6 14 contain have 877:880 arg1 Pofuts 870:875 Pofuts 870:875 Pofuts 870:875 Pofuts have known physiological relevance and are conserved across metazoans. 25186198 6 14 contain have 877:880 arg2 relevance 902:910 known physiological relevance 882:910 known physiological relevance 882:910 Pofuts have known physiological relevance and are conserved across metazoans. 25186198 4 15 theme TSR-containing 711:724 arg1 proteins 726:733 TSR-containing proteins 711:733 TSR-containing proteins 711:733 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 3 16 theme EGF 436:438 arg1 repeats 441:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 8 17 theme past 1159:1162 arg1 advances 1175:1182 past and recent advances 1159:1182 past and recent advances made in novel roles for these protein O-glycosyltransferases 1159:1243 This article discusses past and recent advances made in novel roles for these protein O-glycosyltransferases. 25186198 1 18 from control 121:127 arg1 ER 159:160 ER 159:160 ER 159:160 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 1 18 from control 121:127 arg1 reticulum 148:156 the endoplasmic reticulum 132:156 the endoplasmic reticulum (ER) 132:161 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 4 19 theme folding 680:686 arg1 control 700:706 the folding and quality control 676:706 the folding and quality control of TSR-containing proteins 676:733 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 3 20 theme Growth 416:421 arg1 repeats 441:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 0 21 theme Novel 0:4 arg1 roles 6:10 Novel roles 0:10 Novel roles for O-linked glycans in protein folding 0:50 Novel roles for O-linked glycans in protein folding. 25186198 1 22 from folding 101:107 arg1 ER 159:160 ER 159:160 ER 159:160 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 1 22 from folding 101:107 arg1 reticulum 148:156 the endoplasmic reticulum 132:156 the endoplasmic reticulum (ER) 132:161 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 3 23 theme Factor-like 423:433 arg1 repeats 441:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 0 24 theme O-linked 16:23 arg1 glycans 25:31 O-linked glycans 16:31 O-linked glycans 16:31 Novel roles for O-linked glycans in protein folding. 25186198 5 25 theme unfolded 849:856 arg1 structures 858:867 folded and unfolded structures 838:867 folded and unfolded structures 838:867 Both enzymes only modify fully folded substrates suggesting that they are able to distinguish between folded and unfolded structures. 25186198 4 26 theme quality 692:698 arg1 control 700:706 the folding and quality control 676:706 the folding and quality control of TSR-containing proteins 676:733 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 1 27 theme protein 93:99 arg1 folding 101:107 protein folding 93:107 protein folding 93:107 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 4 28 theme type 608:611 arg1 repeats 615:621 Thrombospondin type I repeats 593:621 Thrombospondin type I repeats (TSRs) 593:628 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 4 28 theme type 608:611 arg1 TSRs 624:627 TSRs 624:627 TSRs 624:627 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 7 29 theme Pofuts 1039:1044 arg1 structures 1020:1029 structures 1020:1029 structures of both Pofuts 1020:1044 Though consensus sequences for O-fucosylation have been established and structures of both Pofuts have been studied, the mechanism of how they participate in protein folding is not known. 25186198 3 30 theme Epidermal 406:414 arg1 repeats 441:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Epidermal Growth Factor-like (EGF) repeats 406:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 1 31 attach linked 83:88 arg1 folding 101:107 protein folding 93:107 protein folding 93:107 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 1 31 attach linked 83:88 arg1 control 121:127 quality control 113:127 quality control 113:127 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 1 31 attach linked 83:88 arg2 N-Glycosylation 53:67 N-Glycosylation 53:67 N-Glycosylation 53:67 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 3 32 theme soluble 360:366 arg1 O-fucosyltransferase 319:338 Protein O-fucosyltransferase 1 311:340 Protein O-fucosyltransferase 1 (Ofut1/Pofut1) 311:355 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 3 32 theme soluble 360:366 arg1 enzyme 382:387 a soluble, ER localized enzyme 358:387 a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats 358:447 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 5 33 theme folded 767:772 arg1 substrates 774:783 fully folded substrates 761:783 fully folded substrates suggesting that they are able to distinguish between folded and unfolded structures 761:867 Both enzymes only modify fully folded substrates suggesting that they are able to distinguish between folded and unfolded structures. 25186198 7 34 theme protein 1106:1112 arg1 folding 1114:1120 protein folding 1106:1120 protein folding 1106:1120 Though consensus sequences for O-fucosylation have been established and structures of both Pofuts have been studied, the mechanism of how they participate in protein folding is not known. 25186198 5 35 theme folded 838:843 arg1 structures 858:867 folded and unfolded structures 838:867 folded and unfolded structures 838:867 Both enzymes only modify fully folded substrates suggesting that they are able to distinguish between folded and unfolded structures. 25186198 2 36 theme O-linked 191:198 arg1 glycosylation 200:212 O-linked glycosylation 191:212 O-linked glycosylation 191:212 Recent work has shown that O-linked glycosylation and the corresponding glycosyltransferases also participate in this important function. 25186198 0 37 theme protein 36:42 arg1 folding 44:50 protein folding 36:50 protein folding 36:50 Novel roles for O-linked glycans in protein folding. 25186198 6 38 theme physiological 888:900 arg1 relevance 902:910 known physiological relevance 882:910 known physiological relevance 882:910 Pofuts have known physiological relevance and are conserved across metazoans. 25186198 8 39 theme protein 1214:1220 arg1 O-glycosyltransferases 1222:1243 these protein O-glycosyltransferases 1208:1243 these protein O-glycosyltransferases 1208:1243 This article discusses past and recent advances made in novel roles for these protein O-glycosyltransferases. 25186198 1 40 theme quality 113:119 arg1 control 121:127 quality control 113:127 quality control 113:127 N-Glycosylation has long been linked to protein folding and quality control in the endoplasmic reticulum (ER). 25186198 2 41 link O-linked 191:198 arg1 glycosylation 200:212 O-linked glycosylation 191:212 O-linked glycosylation 191:212 Recent work has shown that O-linked glycosylation and the corresponding glycosyltransferases also participate in this important function. 25186198 3 42 theme Notch 518:522 arg1 receptor 524:531 the Notch receptor 514:531 the Notch receptor 514:531 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 2 43 theme Recent 164:169 arg1 work 171:174 Recent work 164:174 Recent work 164:174 Recent work has shown that O-linked glycosylation and the corresponding glycosyltransferases also participate in this important function. 25186198 2 44 theme corresponding 222:234 arg1 glycosyltransferases 236:255 the corresponding glycosyltransferases 218:255 the corresponding glycosyltransferases 218:255 Recent work has shown that O-linked glycosylation and the corresponding glycosyltransferases also participate in this important function. 25186198 3 45 theme proper 491:496 arg1 localization 498:509 the proper localization 487:509 the proper localization of the Notch receptor in certain contexts 487:551 Notably, Protein O-fucosyltransferase 1 (Ofut1/Pofut1), a soluble, ER localized enzyme that fucosylates Epidermal Growth Factor-like (EGF) repeats, functions as a chaperone involved in the proper localization of the Notch receptor in certain contexts. 25186198 4 46 theme Thrombospondin 593:606 arg1 type 608:611 Thrombospondin type I 593:613 Thrombospondin type I repeats (TSRs) 593:628 Pofut2, a related enzyme that modifies Thrombospondin type I repeats (TSRs), has also been hypothesized to play a role in the folding and quality control of TSR-containing proteins. 25186198 8 47 theme novel 1192:1196 arg1 roles 1198:1202 novel roles 1192:1202 novel roles for these protein O-glycosyltransferases 1192:1243 This article discusses past and recent advances made in novel roles for these protein O-glycosyltransferases. 25186198 6 48 theme known 882:886 arg1 relevance 902:910 known physiological relevance 882:910 known physiological relevance 882:910 Pofuts have known physiological relevance and are conserved across metazoans. 27791356 7 0 theme atomic 1436:1441 arg1 microscopy 1449:1458 atomic force microscopy 1436:1458 atomic force microscopy 1436:1458 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 6 1 theme region 1100:1105 arg1 proteins 1073:1080 recombinant proteins 1061:1080 recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) 1061:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 3 2 theme glycosylation 541:553 arg1 patterns 555:562 glycosylation patterns 541:562 glycosylation patterns along individual molecules 541:589 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 8 3 theme many 1638:1641 arg1 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 6 4 theme mucin 1237:1241 arg1 domain 1243:1248 the central mucin domain 1225:1248 the central mucin domain of MUC5B 1225:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 4 theme mucin 1237:1241 arg1 MUC5B 1253:1257 MUC5B 1253:1257 MUC5B 1253:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 5 dep domains 1183:1189 arg1 CT5B 1192:1195 CT5B 1192:1195 CT5B 1192:1195 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 1 6 theme polymeric 185:193 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 6 theme polymeric 185:193 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 7 7 theme spectroscopy 1392:1403 arg1 specificity 1371:1381 the chemical specificity 1358:1381 the chemical specificity of Raman spectroscopy 1358:1403 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 6 8 theme C-terminal 1145:1154 arg1 domains 1183:1189 D4-B-C-CK domains 1173:1189 D4-B-C-CK domains 1173:1189 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 8 theme C-terminal 1145:1154 arg1 region 1156:1161 C-terminal region 1145:1161 C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) 1145:1196 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 8 9 from region 1708:1713 arg1 mucin 1722:1726 the mucin 1718:1726 the mucin 1718:1726 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 2 10 theme viscoelastic 322:333 arg1 properties 335:344 their viscoelastic properties 316:344 their viscoelastic properties 316:344 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 6 11 theme Cys-domain 1206:1215 arg1 proteins 1073:1080 recombinant proteins 1061:1080 recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) 1061:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 5 12 theme protein 851:857 arg1 backbone 859:866 the protein backbone 847:866 the protein backbone of MUC5B 847:875 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 4 13 theme tip-enhanced 715:726 arg1 TERS 748:751 TERS 748:751 TERS 748:751 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 13 theme tip-enhanced 715:726 arg1 spectroscopy 734:745 tip-enhanced Raman spectroscopy 715:745 tip-enhanced Raman spectroscopy (TERS) 715:752 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 14 used utilized 627:634 arg2 we 624:625 we 624:625 we 624:625 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 2 15 theme molecular 229:237 arg1 key 309:311 key 309:311 key 309:311 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 2 15 theme molecular 229:237 arg1 weight 239:244 The very high molecular weight 215:244 The very high molecular weight 215:244 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 1 16 theme mucus 132:136 arg1 hydrogels 138:146 protective mucus hydrogels 121:146 protective mucus hydrogels on mucosal surfaces 121:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 6 17 located found 1265:1269 arg1 contrast 1051:1058 sharp contrast 1045:1058 sharp contrast 1045:1058 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 17 located found 1265:1269 arg2 proteins 1073:1080 recombinant proteins 1061:1080 recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) 1061:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 2 18 theme gel-forming 279:289 arg1 mucins 291:296 gel-forming mucins 279:296 gel-forming mucins 279:296 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 8 19 gly O-glycosylated 1693:1706 arg1 region 1708:1713 a highly O-glycosylated region 1684:1713 a highly O-glycosylated region on the mucin 1684:1726 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 0 20 theme Raman 64:68 arg1 Spectroscopies 70:83 Raman Spectroscopies 64:83 Raman Spectroscopies 64:83 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 7 21 theme MUC5B 1522:1526 arg1 molecule 1528:1535 an individual MUC5B molecule 1508:1535 an individual MUC5B molecule 1508:1535 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 8 22 contain contain 1579:1585 arg2 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 22 contain contain 1579:1585 arg2 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 22 contain contain 1579:1585 arg1 molecule 1557:1564 the molecule 1553:1564 the molecule 1553:1564 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 22 contain contain 1579:1585 arg2 amount 1595:1600 a large amount 1587:1600 a large amount of α-helix/unordered structures and many signatures of glycosylation 1587:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 5 23 theme glycosylated 963:974 arg1 domain 990:995 the heavily glycosylated central mucin domain 951:995 the heavily glycosylated central mucin domain 951:995 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 3 24 gly glycosylation 541:553 arg1 subdomains 525:534 the various mucin subdomains 507:534 the various mucin subdomains 507:534 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 1 25 from hydrogels 138:146 arg1 surfaces 159:166 mucosal surfaces 151:166 mucosal surfaces 151:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 5 26 theme mucin 984:988 arg1 domain 990:995 the heavily glycosylated central mucin domain 951:995 the heavily glycosylated central mucin domain 951:995 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 5 27 gly glycosylated 963:974 arg1 domain 990:995 the heavily glycosylated central mucin domain 951:995 the heavily glycosylated central mucin domain 951:995 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 0 28 theme Secondary 0:8 arg1 Structure 10:18 Secondary Structure 0:18 Secondary Structure 0:18 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 1 29 theme structural 96:105 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 29 theme structural 96:105 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 6 30 theme recombinant 1061:1071 arg1 proteins 1073:1080 recombinant proteins 1061:1080 recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) 1061:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 3 31 theme structural 448:457 arg1 information 459:469 key structural information 444:469 key structural information 444:469 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 8 32 theme signatures 1643:1652 arg1 amount 1595:1600 a large amount 1587:1600 a large amount of α-helix/unordered structures and many signatures of glycosylation 1587:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 32 theme signatures 1643:1652 arg1 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 32 theme signatures 1643:1652 arg1 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 4 33 theme MUC5B 821:825 arg1 structure 767:775 the structure 763:775 the structure of the secreted polymeric gel-forming mucin MUC5B 763:825 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 3 34 theme secondary 484:492 arg1 structure 494:502 the secondary structure 480:502 the secondary structure of the various mucin subdomains 480:534 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 5 35 theme O-glycan-rich 1019:1031 arg1 regions 1033:1039 MUC5B O-glycan-rich regions 1013:1039 MUC5B O-glycan-rich regions 1013:1039 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 3 36 theme subdomains 525:534 arg1 structure 494:502 the secondary structure 480:502 the secondary structure of the various mucin subdomains 480:534 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 3 36 theme subdomains 525:534 arg1 patterns 555:562 glycosylation patterns 541:562 glycosylation patterns along individual molecules 541:589 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 0 37 theme Glycoproteins 47:59 arg1 Glycosylation 24:36 Glycosylation 24:36 Glycosylation 24:36 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 0 37 theme Glycoproteins 47:59 arg1 Structure 10:18 Secondary Structure 0:18 Secondary Structure 0:18 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 0 38 gly Glycosylation 24:36 arg1 Glycoproteins 47:59 Mucus Glycoproteins 41:59 Mucus Glycoproteins 41:59 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 4 39 theme secreted 784:791 arg1 MUC5B 821:825 the secreted polymeric gel-forming mucin MUC5B 780:825 the secreted polymeric gel-forming mucin MUC5B 780:825 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 3 40 theme various 511:517 arg1 subdomains 525:534 the various mucin subdomains 507:534 the various mucin subdomains 507:534 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 6 41 theme region 1156:1161 arg1 proteins 1073:1080 recombinant proteins 1061:1080 recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) 1061:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 7 42 theme force 1443:1447 arg1 microscopy 1449:1458 atomic force microscopy 1436:1458 atomic force microscopy 1436:1458 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 4 43 theme Raman 636:640 arg1 spectroscopy 642:653 Raman spectroscopy 636:653 Raman spectroscopy 636:653 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 44 theme gel-forming 803:813 arg1 MUC5B 821:825 the secreted polymeric gel-forming mucin MUC5B 780:825 the secreted polymeric gel-forming mucin MUC5B 780:825 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 2 45 theme conventional 390:401 arg1 techniques 426:435 conventional biochemical/structural techniques 390:435 conventional biochemical/structural techniques 390:435 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 6 46 theme MUC5B 1253:1257 arg1 domain 1243:1248 the central mucin domain 1225:1248 the central mucin domain of MUC5B 1225:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 46 theme MUC5B 1253:1257 arg1 MUC5B 1253:1257 MUC5B 1253:1257 MUC5B 1253:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 8 47 theme α-helix/unordered 1605:1621 arg1 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 7 48 theme spatial 1414:1420 arg1 resolution 1422:1431 the spatial resolution 1410:1431 the spatial resolution of atomic force microscopy 1410:1458 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 6 49 theme N-terminal 1089:1098 arg1 domains 1129:1135 D1-D2-D'-D3 domains 1117:1135 D1-D2-D'-D3 domains 1117:1135 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 49 theme N-terminal 1089:1098 arg1 region 1100:1105 the N-terminal region 1085:1105 the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B) 1085:1142 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 domains 1129:1135 D1-D2-D'-D3 domains 1117:1135 D1-D2-D'-D3 domains 1117:1135 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 region 1156:1161 C-terminal region 1145:1161 C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) 1145:1196 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 MUC5B 1166:1170 MUC5B 1166:1170 MUC5B 1166:1170 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 region 1100:1105 the N-terminal region 1085:1105 the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B) 1085:1142 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 MUC5B 1110:1114 MUC5B 1110:1114 MUC5B 1110:1114 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 Cys-domain 1206:1215 the Cys-domain 1202:1215 the Cys-domain (within the central mucin domain of MUC5B) 1202:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 50 theme MUC5B 1166:1170 arg1 domains 1183:1189 D4-B-C-CK domains 1173:1189 D4-B-C-CK domains 1173:1189 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 51 theme central 1229:1235 arg1 domain 1243:1248 the central mucin domain 1225:1248 the central mucin domain of MUC5B 1225:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 51 theme central 1229:1235 arg1 MUC5B 1253:1257 MUC5B 1253:1257 MUC5B 1253:1257 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 3 52 theme individual 570:579 arg1 molecules 581:589 individual molecules 570:589 individual molecules 570:589 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 1 53 theme gel-forming 195:205 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 53 theme gel-forming 195:205 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 54 theme secreted 176:183 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 54 theme secreted 176:183 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 7 55 theme Raman 1386:1390 arg1 spectroscopy 1392:1403 Raman spectroscopy 1386:1403 Raman spectroscopy 1386:1403 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 7 56 theme chemical 1362:1369 arg1 specificity 1371:1381 the chemical specificity 1358:1381 the chemical specificity of Raman spectroscopy 1358:1403 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 8 57 theme glycosylation 1657:1669 arg1 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 57 theme glycosylation 1657:1669 arg1 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 5 58 theme MUC5B 871:875 arg1 backbone 859:866 the protein backbone 847:866 the protein backbone of MUC5B 847:875 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 4 59 theme circular 686:693 arg1 CD 706:707 CD 706:707 CD 706:707 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 59 theme circular 686:693 arg1 dichroism 695:703 circular dichroism 686:703 circular dichroism (CD) 686:708 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 2 60 gly O-glycosylation 260:274 arg1 mucins 291:296 gel-forming mucins 279:296 gel-forming mucins 279:296 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 4 61 theme optical 662:668 arg1 ROA 680:682 ROA 680:682 ROA 680:682 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 61 theme optical 662:668 arg1 activity 670:677 Raman optical activity 656:677 Raman optical activity (ROA) 656:683 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 2 62 theme extensive 250:258 arg1 O-glycosylation 260:274 extensive O-glycosylation 250:274 extensive O-glycosylation of gel-forming mucins 250:296 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 8 63 theme large 1589:1593 arg1 amount 1595:1600 a large amount 1587:1600 a large amount of α-helix/unordered structures and many signatures of glycosylation 1587:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 63 theme large 1589:1593 arg1 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 63 theme large 1589:1593 arg1 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 64 theme O-glycosylated 1693:1706 arg1 region 1708:1713 a highly O-glycosylated region 1684:1713 a highly O-glycosylated region on the mucin 1684:1726 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 1 65 theme protective 121:130 arg1 hydrogels 138:146 protective mucus hydrogels 121:146 protective mucus hydrogels on mucosal surfaces 121:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 2 66 theme mucins 291:296 arg1 key 309:311 key 309:311 key 309:311 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 2 66 theme mucins 291:296 arg1 weight 239:244 The very high molecular weight 215:244 The very high molecular weight 215:244 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 2 66 theme mucins 291:296 arg1 O-glycosylation 260:274 extensive O-glycosylation 250:274 extensive O-glycosylation of gel-forming mucins 250:296 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 1 67 theme hydrogels 138:146 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 67 theme hydrogels 138:146 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 7 68 theme molecule 1528:1535 arg1 structure 1483:1491 the secondary structure 1469:1491 the secondary structure along 90 nm of an individual MUC5B molecule 1469:1535 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 4 69 theme Raman 728:732 arg1 TERS 748:751 TERS 748:751 TERS 748:751 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 69 theme Raman 728:732 arg1 spectroscopy 734:745 tip-enhanced Raman spectroscopy 715:745 tip-enhanced Raman spectroscopy (TERS) 715:752 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 1 70 theme mucosal 151:157 arg1 surfaces 159:166 mucosal surfaces 151:166 mucosal surfaces 151:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 7 71 theme individual 1511:1520 arg1 molecule 1528:1535 an individual MUC5B molecule 1508:1535 an individual MUC5B molecule 1508:1535 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 6 72 theme D4-B-C-CK 1173:1181 arg1 domains 1183:1189 D4-B-C-CK domains 1173:1189 D4-B-C-CK domains 1173:1189 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 72 theme D4-B-C-CK 1173:1181 arg1 region 1156:1161 C-terminal region 1145:1161 C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) 1145:1196 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 5 73 theme unordered 893:901 arg1 conformation 903:914 unordered conformation 893:914 unordered conformation 893:914 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 1 74 from surfaces 159:166 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 74 from surfaces 159:166 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 5 75 theme central 976:982 arg1 domain 990:995 the heavily glycosylated central mucin domain 951:995 the heavily glycosylated central mucin domain 951:995 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 1 76 theme major 90:94 arg1 mucins 207:212 the secreted polymeric gel-forming mucins 172:212 the secreted polymeric gel-forming mucins 172:212 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 1 76 theme major 90:94 arg1 components 107:116 The major structural components 86:116 The major structural components of protective mucus hydrogels on mucosal surfaces 86:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 2 77 theme biochemical/structural 403:424 arg1 techniques 426:435 conventional biochemical/structural techniques 390:435 conventional biochemical/structural techniques 390:435 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 0 78 theme Mucus 41:45 arg1 Glycoproteins 47:59 Mucus Glycoproteins 41:59 Mucus Glycoproteins 41:59 Secondary Structure and Glycosylation of Mucus Glycoproteins by Raman Spectroscopies. 27791356 5 79 theme MUC5B 1013:1017 arg1 regions 1033:1039 MUC5B O-glycan-rich regions 1013:1039 MUC5B O-glycan-rich regions 1013:1039 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 6 80 theme sharp 1045:1049 arg1 contrast 1051:1058 sharp contrast 1045:1058 sharp contrast 1045:1058 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 4 81 theme mucin 815:819 arg1 MUC5B 821:825 the secreted polymeric gel-forming mucin MUC5B 780:825 the secreted polymeric gel-forming mucin MUC5B 780:825 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 7 82 theme secondary 1473:1481 arg1 structure 1483:1491 the secondary structure 1469:1491 the secondary structure along 90 nm of an individual MUC5B molecule 1469:1535 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 5 83 theme regions 1033:1039 arg1 isolation 1000:1008 isolation 1000:1008 isolation of MUC5B O-glycan-rich regions 1000:1039 ROA indicated that the protein backbone of MUC5B is dominated by unordered conformation, which was found to originate from the heavily glycosylated central mucin domain by isolation of MUC5B O-glycan-rich regions. 27791356 3 84 theme key 444:446 arg1 information 459:469 key structural information 444:469 key structural information 444:469 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 2 85 theme high 224:227 arg1 key 309:311 key 309:311 key 309:311 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 2 85 theme high 224:227 arg1 weight 239:244 The very high molecular weight 215:244 The very high molecular weight 215:244 The very high molecular weight and extensive O-glycosylation of gel-forming mucins, which are key to their viscoelastic properties, create problems when studying mucins using conventional biochemical/structural techniques. 27791356 1 86 from components 107:116 arg1 surfaces 159:166 mucosal surfaces 151:166 mucosal surfaces 151:166 The major structural components of protective mucus hydrogels on mucosal surfaces are the secreted polymeric gel-forming mucins. 27791356 4 87 theme polymeric 793:801 arg1 MUC5B 821:825 the secreted polymeric gel-forming mucin MUC5B 780:825 the secreted polymeric gel-forming mucin MUC5B 780:825 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 6 88 theme D1-D2-D'-D3 1117:1127 arg1 domains 1129:1135 D1-D2-D'-D3 domains 1117:1135 D1-D2-D'-D3 domains 1117:1135 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 88 theme D1-D2-D'-D3 1117:1127 arg1 region 1100:1105 the N-terminal region 1085:1105 the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B) 1085:1142 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 3 89 theme mucin 519:523 arg1 subdomains 525:534 the various mucin subdomains 507:534 the various mucin subdomains 507:534 Thus, key structural information, such as the secondary structure of the various mucin subdomains, and glycosylation patterns along individual molecules, remains to be elucidated. 27791356 7 90 theme microscopy 1449:1458 arg1 resolution 1422:1431 the spatial resolution 1410:1431 the spatial resolution of atomic force microscopy 1410:1458 Using these findings, we employed TERS, which combines the chemical specificity of Raman spectroscopy with the spatial resolution of atomic force microscopy to study the secondary structure along 90 nm of an individual MUC5B molecule. 27791356 6 91 theme MUC5B 1110:1114 arg1 domains 1129:1135 D1-D2-D'-D3 domains 1117:1135 D1-D2-D'-D3 domains 1117:1135 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 region 1156:1161 C-terminal region 1145:1161 C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) 1145:1196 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 MUC5B 1166:1170 MUC5B 1166:1170 MUC5B 1166:1170 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 region 1100:1105 the N-terminal region 1085:1105 the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B) 1085:1142 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 MUC5B 1110:1114 MUC5B 1110:1114 MUC5B 1110:1114 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 Cys-domain 1206:1215 the Cys-domain 1202:1215 the Cys-domain (within the central mucin domain of MUC5B) 1202:1258 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 6 91 theme MUC5B 1110:1114 arg1 domains 1183:1189 D4-B-C-CK domains 1173:1189 D4-B-C-CK domains 1173:1189 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 4 92 theme Raman 656:660 arg1 ROA 680:682 ROA 680:682 ROA 680:682 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 4 92 theme Raman 656:660 arg1 activity 670:677 Raman optical activity 656:677 Raman optical activity (ROA) 656:683 Here, we utilized Raman spectroscopy, Raman optical activity (ROA), circular dichroism (CD), and tip-enhanced Raman spectroscopy (TERS) to study the structure of the secreted polymeric gel-forming mucin MUC5B. 27791356 6 93 dep domains 1129:1135 arg1 NT5B 1138:1141 NT5B 1138:1141 NT5B 1138:1141 In sharp contrast, recombinant proteins of the N-terminal region of MUC5B (D1-D2-D'-D3 domains, NT5B), C-terminal region of MUC5B (D4-B-C-CK domains, CT5B) and the Cys-domain (within the central mucin domain of MUC5B) were found to be dominated by the β-sheet. 27791356 8 94 theme structures 1623:1632 arg1 amount 1595:1600 a large amount 1587:1600 a large amount of α-helix/unordered structures and many signatures of glycosylation 1587:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 94 theme structures 1623:1632 arg1 structures 1623:1632 α-helix/unordered structures 1605:1632 α-helix/unordered structures 1605:1632 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 27791356 8 94 theme structures 1623:1632 arg1 signatures 1643:1652 many signatures 1638:1652 many signatures of glycosylation 1638:1669 Interestingly, the molecule was found to contain a large amount of α-helix/unordered structures and many signatures of glycosylation, pointing to a highly O-glycosylated region on the mucin. 25496419 0 0 theme disaccharide 57:68 arg1 synthesis 26:34 First and stereoselective synthesis 0:34 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 0:114 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 4 1 theme inner 694:698 arg1 structure 705:713 this unique bacterial Kdo inner core structure 668:713 this unique bacterial Kdo inner core structure 668:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 1 2 theme pathogens 140:148 arg1 Resistance 116:125 Resistance 116:125 Resistance of bacterial pathogens toward antibiotics 116:167 Resistance of bacterial pathogens toward antibiotics has revived interest in lipopolysaccharide (LPS) motifs as potential therapeutic targets. 25496419 4 3 theme linkage 585:591 arg1 type 593:596 this linkage type 580:596 this linkage type 580:596 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 0 4 theme -linked 49:55 arg1 disaccharide 57:68 an α-(2→5)-linked disaccharide 39:68 an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) 39:113 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 3 5 theme good 511:514 arg1 yield 516:520 good yield 511:520 good yield 511:520 In this contribution the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield is highlighted. 25496419 0 6 theme 3-deoxy-D-manno-oct-2-ulosonic 73:102 arg1 Kdo 110:112 Kdo 110:112 Kdo 110:112 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 0 6 theme 3-deoxy-D-manno-oct-2-ulosonic 73:102 arg1 acid 104:107 3-deoxy-D-manno-oct-2-ulosonic acid 73:107 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) 73:113 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 2 7 contain containing 354:363 arg1 trisaccharide 340:352 a 4,5-branched Kdo trisaccharide 321:352 a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage 321:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 2 7 contain containing 354:363 arg2 -linkage 382:389 an uncommon (2→5)-linkage 365:389 an uncommon (2→5)-linkage 365:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 2 8 theme pathogenic 278:287 arg1 strains 303:309 several pathogenic Acinetobacter strains 270:309 several pathogenic Acinetobacter strains 270:309 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 2 9 theme several 270:276 arg1 strains 303:309 several pathogenic Acinetobacter strains 270:309 several pathogenic Acinetobacter strains 270:309 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 4 10 theme core 700:703 arg1 structure 705:713 this unique bacterial Kdo inner core structure 668:713 this unique bacterial Kdo inner core structure 668:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 4 11 theme bacterial 680:688 arg1 structure 705:713 this unique bacterial Kdo inner core structure 668:713 this unique bacterial Kdo inner core structure 668:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 0 12 theme acid 104:107 arg1 disaccharide 57:68 an α-(2→5)-linked disaccharide 39:68 an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) 39:113 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 3 13 theme stereoselective 423:437 arg1 method 453:458 the first stereoselective glycosylation method 413:458 the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield 413:520 In this contribution the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield is highlighted. 25496419 0 14 theme First 0:4 arg1 synthesis 26:34 First and stereoselective synthesis 0:34 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 0:114 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 3 15 theme glycosylation 439:451 arg1 method 453:458 the first stereoselective glycosylation method 413:458 the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield 413:520 In this contribution the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield is highlighted. 25496419 2 16 theme strains 303:309 arg1 LPS 263:265 The LPS 259:265 The LPS of several pathogenic Acinetobacter strains 259:309 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 0 17 theme stereoselective 10:24 arg1 synthesis 26:34 First and stereoselective synthesis 0:34 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 0:114 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 2 18 theme Acinetobacter 289:301 arg1 strains 303:309 several pathogenic Acinetobacter strains 270:309 several pathogenic Acinetobacter strains 270:309 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 1 19 theme lipopolysaccharide 193:210 arg1 motifs 218:223 lipopolysaccharide (LPS) motifs 193:223 lipopolysaccharide (LPS) motifs as potential therapeutic targets 193:256 Resistance of bacterial pathogens toward antibiotics has revived interest in lipopolysaccharide (LPS) motifs as potential therapeutic targets. 25496419 1 20 dep potential 228:236 arg1 therapeutic 238:248 therapeutic 238:248 therapeutic 238:248 Resistance of bacterial pathogens toward antibiotics has revived interest in lipopolysaccharide (LPS) motifs as potential therapeutic targets. 25496419 4 21 theme unique 673:678 arg1 structure 705:713 this unique bacterial Kdo inner core structure 668:713 this unique bacterial Kdo inner core structure 668:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 3 22 theme first 417:421 arg1 method 453:458 the first stereoselective glycosylation method 413:458 the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield 413:520 In this contribution the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield is highlighted. 25496419 2 23 theme uncommon 368:375 arg1 -linkage 382:389 an uncommon (2→5)-linkage 365:389 an uncommon (2→5)-linkage 365:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 4 24 theme immunoreactivity 635:650 arg1 studies 620:626 future studies 613:626 future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure 613:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 0 25 link -linked 49:55 arg1 disaccharide 57:68 an α-(2→5)-linked disaccharide 39:68 an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) 39:113 First and stereoselective synthesis of an α-(2→5)-linked disaccharide of 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo). 25496419 3 26 theme -α-Kdo 488:493 arg1 disaccharide 495:506 an α-Kdo-(2→5)-α-Kdo disaccharide 474:506 an α-Kdo-(2→5)-α-Kdo disaccharide 474:506 In this contribution the first stereoselective glycosylation method for obtaining an α-Kdo-(2→5)-α-Kdo disaccharide in good yield is highlighted. 25496419 4 27 theme synthetic 542:550 arg1 approach 552:559 The synthetic approach 538:559 The synthetic approach used for accessing this linkage type 538:596 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 4 28 theme Kdo 690:692 arg1 structure 705:713 this unique bacterial Kdo inner core structure 668:713 this unique bacterial Kdo inner core structure 668:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 2 29 theme Kdo 336:338 arg1 trisaccharide 340:352 a 4,5-branched Kdo trisaccharide 321:352 a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage 321:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 2 30 theme 4,5-branched 323:334 arg1 trisaccharide 340:352 a 4,5-branched Kdo trisaccharide 321:352 a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage 321:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 2 31 theme 2→5 378:380 arg1 -linkage 382:389 an uncommon (2→5)-linkage 365:389 an uncommon (2→5)-linkage 365:389 The LPS of several pathogenic Acinetobacter strains comprises a 4,5-branched Kdo trisaccharide containing an uncommon (2→5)-linkage. 25496419 4 32 theme future 613:618 arg1 studies 620:626 future studies 613:626 future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure 613:713 The synthetic approach used for accessing this linkage type will allow for future studies of the immunoreactivity associated with this unique bacterial Kdo inner core structure. 25496419 1 33 theme bacterial 130:138 arg1 pathogens 140:148 bacterial pathogens 130:148 bacterial pathogens 130:148 Resistance of bacterial pathogens toward antibiotics has revived interest in lipopolysaccharide (LPS) motifs as potential therapeutic targets. 25496419 1 34 theme potential 228:236 arg1 targets 250:256 potential therapeutic targets 228:256 potential therapeutic targets 228:256 Resistance of bacterial pathogens toward antibiotics has revived interest in lipopolysaccharide (LPS) motifs as potential therapeutic targets. 27845725 0 0 theme Anticoagulant 73:85 arg1 Idraparinux 103:113 the Anticoagulant Pentasaccharide Idraparinux 69:113 the Anticoagulant Pentasaccharide Idraparinux 69:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 3 1 theme ongoing 351:357 arg1 research 359:366 our ongoing research 347:366 our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants 347:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 5 2 theme sulfonic 878:885 arg1 end-products 892:903 the fully O-sulfated, O-methylated sulfonic acid end-products 843:903 end-products 892:903 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 3 3 theme primary 557:563 arg1 moieties 579:586 one or two primary sulfonic acid moieties 546:586 one or two primary sulfonic acid moieties 546:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 3 4 theme research 359:366 arg1 frame 338:342 the frame 334:342 the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants 334:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 3 5 theme sulfonic 565:572 arg1 moieties 579:586 one or two primary sulfonic acid moieties 546:586 one or two primary sulfonic acid moieties 546:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 0 6 theme Idraparinux 103:113 arg1 Analogues 56:64 Isosteric Sulfonic Acid Analogues 32:64 Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 32:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 0 7 theme Pentasaccharide 87:101 arg1 Idraparinux 103:113 the Anticoagulant Pentasaccharide Idraparinux 69:113 the Anticoagulant Pentasaccharide Idraparinux 69:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 3 8 theme pentasaccharides 521:536 arg1 series 490:495 a series 488:495 a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties 488:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 5 9 theme O-sulfated 853:862 arg1 end-products 892:903 the fully O-sulfated, O-methylated sulfonic acid end-products 843:903 end-products 892:903 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 1 10 from drugs 149:153 arg1 prophylaxis 184:194 prophylaxis 184:194 prophylaxis 184:194 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 10 from drugs 149:153 arg1 therapy 172:178 therapy 172:178 therapy 172:178 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 2 11 theme antithrombin-binding 302:321 arg1 domain 323:328 the heparin antithrombin-binding domain 290:328 the heparin antithrombin-binding domain 290:328 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 0 12 theme Modular 2:8 arg1 Approach 20:27 A Modular Synthetic Approach 0:27 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 0:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 5 13 theme acid 887:890 arg1 end-products 892:903 the fully O-sulfated, O-methylated sulfonic acid end-products 843:903 end-products 892:903 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 4 14 with pentasaccharides 604:619 arg1 patterns 650:657 different C-sulfonation patterns 626:657 different C-sulfonation patterns 626:657 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 4 15 theme disaccharide 764:775 arg1 blocks 786:791 two monosaccharide and four disaccharide building blocks 736:791 blocks 786:791 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 4 16 theme different 626:634 arg1 patterns 650:657 different C-sulfonation patterns 626:657 different C-sulfonation patterns 626:657 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 1 17 dep therapy 172:178 arg1 the 168:170 the 168:170 the 168:170 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 18 theme thromboembolic 199:212 arg1 diseases 214:221 thromboembolic diseases 199:221 thromboembolic diseases 199:221 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 3 19 theme acid-containing 403:417 arg1 anticoagulants 430:443 sulfonic acid-containing heparinoid anticoagulants 394:443 sulfonic acid-containing heparinoid anticoagulants 394:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 1 20 theme diseases 214:221 arg1 prophylaxis 184:194 prophylaxis 184:194 prophylaxis 184:194 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 20 theme diseases 214:221 arg1 therapy 172:178 therapy 172:178 therapy 172:178 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 3 21 theme heparinoid 419:428 arg1 anticoagulants 430:443 sulfonic acid-containing heparinoid anticoagulants 394:443 sulfonic acid-containing heparinoid anticoagulants 394:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 1 22 theme Heparin-based 116:128 arg1 drugs 149:153 drugs 149:153 drugs of choice 149:163 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 22 theme Heparin-based 116:128 arg1 anticoagulants 130:143 Heparin-based anticoagulants 116:143 Heparin-based anticoagulants 116:143 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 23 theme choice 158:163 arg1 drugs 149:153 drugs 149:153 drugs of choice 149:163 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 1 23 theme choice 158:163 arg1 anticoagulants 130:143 Heparin-based anticoagulants 116:143 Heparin-based anticoagulants 116:143 Heparin-based anticoagulants are drugs of choice in the therapy and prophylaxis of thromboembolic diseases. 27845725 0 24 theme Isosteric 32:40 arg1 Analogues 56:64 Isosteric Sulfonic Acid Analogues 32:64 Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 32:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 4 25 theme C-sulfonation 636:648 arg1 patterns 650:657 different C-sulfonation patterns 626:657 different C-sulfonation patterns 626:657 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 5 26 theme protected 816:824 arg1 derivatives 826:836 the protected derivatives 812:836 the protected derivatives 812:836 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 3 27 theme modular 462:468 arg1 pathway 470:476 a modular pathway 460:476 a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties 460:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 5 28 theme derivatives 826:836 arg1 Transformation 794:807 Transformation 794:807 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products 794:903 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 2 29 theme anticoagulant 251:263 arg1 pentasaccharide 265:279 a synthetic anticoagulant pentasaccharide 239:279 a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain 239:328 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 2 29 theme anticoagulant 251:263 arg1 Idraparinux 224:234 Idraparinux 224:234 Idraparinux 224:234 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 0 30 theme Acid 51:54 arg1 Analogues 56:64 Isosteric Sulfonic Acid Analogues 32:64 Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 32:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 2 31 theme synthetic 241:249 arg1 pentasaccharide 265:279 a synthetic anticoagulant pentasaccharide 239:279 a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain 239:328 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 2 31 theme synthetic 241:249 arg1 Idraparinux 224:234 Idraparinux 224:234 Idraparinux 224:234 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 0 32 theme Sulfonic 42:49 arg1 Analogues 56:64 Isosteric Sulfonic Acid Analogues 32:64 Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 32:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 4 33 theme protected 594:602 arg1 pentasaccharides 604:619 Five protected pentasaccharides 589:619 Five protected pentasaccharides with different C-sulfonation patterns 589:657 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 2 34 theme heparin 294:300 arg1 domain 323:328 the heparin antithrombin-binding domain 290:328 the heparin antithrombin-binding domain 290:328 Idraparinux is a synthetic anticoagulant pentasaccharide based on the heparin antithrombin-binding domain. 27845725 4 35 theme glycosylation 691:703 arg1 reactions 705:713 two subsequent glycosylation reactions 676:713 two subsequent glycosylation reactions 676:713 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 4 36 theme building 777:784 arg1 blocks 786:791 two monosaccharide and four disaccharide building blocks 736:791 blocks 786:791 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 4 37 theme subsequent 680:689 arg1 reactions 705:713 two subsequent glycosylation reactions 676:713 two subsequent glycosylation reactions 676:713 Five protected pentasaccharides with different C-sulfonation patterns were prepared by two subsequent glycosylation reactions, respectively, using two monosaccharide and four disaccharide building blocks. 27845725 3 38 theme idraparinux-analogue 500:519 arg1 pentasaccharides 521:536 idraparinux-analogue pentasaccharides 500:536 idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties 500:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 3 39 theme acid 574:577 arg1 moieties 579:586 one or two primary sulfonic acid moieties 546:586 one or two primary sulfonic acid moieties 546:586 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 5 40 theme O-methylated 865:876 arg1 end-products 892:903 the fully O-sulfated, O-methylated sulfonic acid end-products 843:903 end-products 892:903 Transformation of the protected derivatives into the fully O-sulfated, O-methylated sulfonic acid end-products was also studied. 27845725 0 41 theme Synthetic 10:18 arg1 Approach 20:27 A Modular Synthetic Approach 0:27 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux 0:113 A Modular Synthetic Approach to Isosteric Sulfonic Acid Analogues of the Anticoagulant Pentasaccharide Idraparinux. 27845725 3 42 theme anticoagulants 430:443 arg1 synthesis 381:389 the synthesis 377:389 the synthesis of sulfonic acid-containing heparinoid anticoagulants 377:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 27845725 3 43 theme sulfonic 394:401 arg1 anticoagulants 430:443 sulfonic acid-containing heparinoid anticoagulants 394:443 sulfonic acid-containing heparinoid anticoagulants 394:443 In the frame of our ongoing research aimed at the synthesis of sulfonic acid-containing heparinoid anticoagulants, we elaborated a modular pathway to obtain a series of idraparinux-analogue pentasaccharides bearing one or two primary sulfonic acid moieties. 25868252 0 0 theme recombinant 70:80 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 2 1 theme recombinant 345:355 arg1 protein 357:363 recombinant protein SMB 345:367 recombinant protein SMB(PRG4) 345:373 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 2 1 theme recombinant 345:355 arg1 PRG4 369:372 PRG4 369:372 PRG4 369:372 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 4 2 theme non-reducing 582:593 arg1 electrophoresis 595:609 non-reducing electrophoresis 582:609 non-reducing electrophoresis 582:609 Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer. 25868252 1 3 theme proteoglycan 234:245 arg1 sequences 221:229 repetitive sequences 210:229 repetitive sequences of proteoglycan 4 210:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 0 4 theme aggregation 90:100 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 4 theme aggregation 90:100 arg1 proteoglycan 51:62 proteoglycan 4 51:64 proteoglycan 4 51:64 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 4 theme aggregation 90:100 arg1 domain 41:46 somatomedin b domain 27:46 somatomedin b domain of proteoglycan 4 and recombinant protein aggregation 27:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 1 5 gly glycosylation 193:205 arg1 sequences 221:229 repetitive sequences 210:229 repetitive sequences of proteoglycan 4 210:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 4 6 theme bonded 657:662 arg1 SMB 625:627 SMB 625:627 SMB(PRG4) 625:633 Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer. 25868252 4 6 theme bonded 657:662 arg1 dimer 664:668 a non-covalently bonded dimer 640:668 a non-covalently bonded dimer 640:668 Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer. 25868252 0 7 theme protein 82:88 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 3 8 theme dimeric 543:549 arg1 form 551:554 dimeric form 543:554 dimeric form 543:554 By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. 25868252 1 9 contain containing 134:143 arg1 protein 116:122 Recombinant protein SMB 104:126 Recombinant protein SMB(PRG4) containing two Somatomedin B domains 104:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 9 contain containing 134:143 arg1 PRG4 128:131 PRG4 128:131 PRG4 128:131 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 9 contain containing 134:143 arg2 domains 163:169 two Somatomedin B domains 145:169 two Somatomedin B domains 145:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 3 10 theme dynamic 461:467 arg1 scattering 475:484 dynamic light scattering 461:484 dynamic light scattering 461:484 By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. 25868252 1 11 theme Somatomedin 149:159 arg1 domains 163:169 two Somatomedin B domains 145:169 two Somatomedin B domains 145:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 12 theme B 161:161 arg1 domains 163:169 two Somatomedin B domains 145:169 two Somatomedin B domains 145:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 4 13 theme Structure 557:565 arg1 prediction 567:576 Structure prediction 557:576 Structure prediction 557:576 Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer. 25868252 2 14 theme Mature 297:302 arg1 process 317:323 Mature purification process 297:323 Mature purification process 297:323 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 1 15 theme small 177:181 arg1 glycosylation 193:205 glycosylation 193:205 glycosylation of repetitive sequences of proteoglycan 4 193:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 15 theme small 177:181 arg1 amount 183:188 a small amount 175:188 a small amount of glycosylation of repetitive sequences of proteoglycan 4 175:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 0 16 theme b 39:39 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 16 theme b 39:39 arg1 proteoglycan 51:62 proteoglycan 4 51:64 proteoglycan 4 51:64 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 16 theme b 39:39 arg1 domain 41:46 somatomedin b domain 27:46 somatomedin b domain of proteoglycan 4 and recombinant protein aggregation 27:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 1 17 theme PGR4 273:276 arg1 polymorphism 283:294 PGR4 gene polymorphism 273:294 PGR4 gene polymorphism 273:294 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 3 18 theme size-exclusion 426:439 arg1 chromatogaraphy 441:455 size-exclusion chromatogaraphy 426:455 size-exclusion chromatogaraphy 426:455 By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. 25868252 2 19 theme high-level 381:390 arg1 expression 392:401 high-level expression 381:401 high-level expression 381:401 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 0 20 theme somatomedin 27:37 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 20 theme somatomedin 27:37 arg1 proteoglycan 51:62 proteoglycan 4 51:64 proteoglycan 4 51:64 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 20 theme somatomedin 27:37 arg1 domain 41:46 somatomedin b domain 27:46 somatomedin b domain of proteoglycan 4 and recombinant protein aggregation 27:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 1 21 theme gene 278:281 arg1 polymorphism 283:294 PGR4 gene polymorphism 273:294 PGR4 gene polymorphism 273:294 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 22 theme Recombinant 104:114 arg1 protein 116:122 Recombinant protein SMB 104:126 Recombinant protein SMB(PRG4) containing two Somatomedin B domains 104:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 22 theme Recombinant 104:114 arg1 PRG4 128:131 PRG4 128:131 PRG4 128:131 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 0 23 theme domain 41:46 arg1 expression 13:22 expression 13:22 expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation 13:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 3 24 theme light 469:473 arg1 scattering 475:484 dynamic light scattering 461:484 dynamic light scattering 461:484 By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. 25868252 1 25 theme glycosylation 193:205 arg1 glycosylation 193:205 glycosylation 193:205 glycosylation of repetitive sequences of proteoglycan 4 193:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 25 theme glycosylation 193:205 arg1 amount 183:188 a small amount 175:188 a small amount of glycosylation of repetitive sequences of proteoglycan 4 175:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 25 theme glycosylation 193:205 arg1 protein 116:122 Recombinant protein SMB 104:126 Recombinant protein SMB(PRG4) containing two Somatomedin B domains 104:169 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 1 25 theme glycosylation 193:205 arg1 PRG4 128:131 PRG4 128:131 PRG4 128:131 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 2 26 theme purification 304:315 arg1 process 317:323 Mature purification process 297:323 Mature purification process 297:323 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 2 27 with protein 357:363 arg1 expression 392:401 high-level expression 381:401 high-level expression 381:401 Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. 25868252 1 28 theme repetitive 210:219 arg1 sequences 221:229 repetitive sequences 210:229 repetitive sequences of proteoglycan 4 210:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 25868252 0 29 theme proteoglycan 51:62 arg1 aggregation 90:100 recombinant protein aggregation 70:100 recombinant protein aggregation 70:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 29 theme proteoglycan 51:62 arg1 proteoglycan 51:62 proteoglycan 4 51:64 proteoglycan 4 51:64 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 0 29 theme proteoglycan 51:62 arg1 domain 41:46 somatomedin b domain 27:46 somatomedin b domain of proteoglycan 4 and recombinant protein aggregation 27:100 [Research on expression of somatomedin b domain of proteoglycan 4 and recombinant protein aggregation]. 25868252 3 30 theme recombinant 505:515 arg1 protein 517:523 the recombinant protein 501:523 the recombinant protein 501:523 By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. 25868252 1 31 theme sequences 221:229 arg1 glycosylation 193:205 glycosylation 193:205 glycosylation of repetitive sequences of proteoglycan 4 193:247 Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. 24550396 7 0 dep in 1273:1274 arg1 vitro 1276:1280 vitro 1276:1280 vitro 1276:1280 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 6 1 theme " 1169:1169 arg1 glycans 1171:1177 atypical "pseudohybrid" glycans 1147:1177 atypical "pseudohybrid" glycans 1147:1177 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 8 2 theme β1-2 1637:1640 arg1 linkages 1642:1649 β1-2 linkages 1637:1649 β1-2 linkages 1637:1649 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 4 3 theme genes 678:682 arg1 TbGT11 685:690 TbGT11 685:690 TbGT11 685:690 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 3 theme genes 678:682 arg1 one 665:667 one 665:667 one 665:667 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 3 theme genes 678:682 arg1 genes 678:682 these genes 672:682 these genes 672:682 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 5 4 theme modified 893:900 arg1 N-glycans 910:918 significantly modified protein N-glycans 879:918 significantly modified protein N-glycans 879:918 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 2 5 theme β1-2- 392:396 arg1 genes 438:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 0 6 from Identification 0:13 arg1 brucei 127:132 Trypanosoma brucei 115:132 Trypanosoma brucei 115:132 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 2 7 theme latter 323:328 arg1 presence 307:314 the presence 303:314 the presence of the latter 303:328 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 8 8 theme T. 1486:1487 arg1 enzyme 1496:1501 the T. brucei enzyme 1482:1501 the T. brucei enzyme 1482:1501 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 6 9 theme T. 1107:1108 arg1 mutants 1129:1135 T. brucei TbGT11 null mutants 1107:1135 T. brucei TbGT11 null mutants 1107:1135 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 5 10 theme bloodstream-form 833:848 arg1 mutant 862:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 6 11 theme GnTI 1025:1028 arg1 reaction 1030:1037 the GnTI reaction 1021:1037 the GnTI reaction 1021:1037 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 6 11 theme GnTI 1025:1028 arg1 essential 1042:1050 essential 1042:1050 essential 1042:1050 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 3 12 theme UDP-sugar-dependent 537:555 arg1 glycosyltransferases 557:576 putative UDP-sugar-dependent glycosyltransferases 528:576 putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family 528:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 5 13 theme TbGT11 850:855 arg1 mutant 862:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 6 14 theme TbGnTII 1196:1202 arg1 activity 1204:1211 TbGnTII activity 1196:1211 TbGnTII activity 1196:1211 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 3 15 theme β1-3-glycosyltransferase 611:634 arg1 family 636:641 the mammalian β1-3-glycosyltransferase family 597:641 the mammalian β1-3-glycosyltransferase family 597:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 1 16 theme N-glycans 188:196 arg1 N-glycans 188:196 N-glycans 188:196 N-glycans 188:196 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 1 16 theme N-glycans 188:196 arg1 repertoire 174:183 a diverse repertoire 164:183 a diverse repertoire 164:183 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 0 17 theme Trypanosoma 115:125 arg1 brucei 127:132 Trypanosoma brucei 115:132 Trypanosoma brucei 115:132 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 6 18 theme TbGT11 1117:1122 arg1 mutants 1129:1135 T. brucei TbGT11 null mutants 1107:1135 T. brucei TbGT11 null mutants 1107:1135 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 7 19 theme preferred 1410:1418 arg1 Man5GlcNAc2 1384:1394 triantennary Man5GlcNAc2 1371:1394 triantennary Man5GlcNAc2 1371:1394 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 7 19 theme preferred 1410:1418 arg1 substrate 1420:1428 the preferred substrate 1406:1428 the preferred substrate for metazoan GnTIs 1406:1447 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 6 20 dep organisms 1004:1012 arg1 contrast 978:985 contrast 978:985 contrast 978:985 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 4 21 theme apparatus 726:734 arg1 it 707:708 it 707:708 it 707:708 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 21 theme apparatus 726:734 arg1 UDP-GlcNAc 745:754 a Golgi apparatus resident UDP-GlcNAc 718:754 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 3 22 theme mammalian 601:609 arg1 family 636:641 the mammalian β1-3-glycosyltransferase family 597:641 the mammalian β1-3-glycosyltransferase family 597:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 3 23 with glycosyltransferases 557:576 arg1 similarity 583:592 similarity 583:592 similarity to the mammalian β1-3-glycosyltransferase family 583:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 7 24 theme triantennary 1371:1382 arg1 Man5GlcNAc2 1384:1394 triantennary Man5GlcNAc2 1371:1394 triantennary Man5GlcNAc2 1371:1394 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 7 24 theme triantennary 1371:1382 arg1 substrate 1420:1428 the preferred substrate 1406:1428 the preferred substrate for metazoan GnTIs 1406:1447 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 6 25 theme N-glycans 1096:1104 arg1 biosynthesis 1056:1067 biosynthesis 1056:1067 biosynthesis of both complex and hybrid N-glycans 1056:1104 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 2 26 theme obvious 334:340 arg1 homologues 342:351 no obvious homologues 331:351 no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 331:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 7 27 theme in 1273:1274 arg1 assay 1282:1286 a functional in vitro assay 1260:1286 a functional in vitro assay 1260:1286 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 6 28 theme hybrid 1089:1094 arg1 N-glycans 1096:1104 hybrid N-glycans 1089:1104 hybrid N-glycans 1089:1104 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 6 29 theme pseudohybrid 1157:1168 arg1 glycans 1171:1177 atypical "pseudohybrid" glycans 1147:1177 atypical "pseudohybrid" glycans 1147:1177 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 8 30 theme β3-glycosyltransferase 1595:1616 arg1 family 1618:1623 the β3-glycosyltransferase family 1591:1623 the β3-glycosyltransferase family 1591:1623 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 1 31 theme oligomannose 212:223 arg1 structures 242:251 oligomannose and paucimannose structures 212:251 structures 242:251 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 0 32 theme functional 19:28 arg1 characterization 30:45 functional characterization 19:45 functional characterization 19:45 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 6 33 theme multicellular 990:1002 arg1 organisms 1004:1012 multicellular organisms 990:1012 multicellular organisms 990:1012 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 4 34 theme I 808:808 arg1 TbGnTI 820:825 TbGnTI 820:825 TbGnTI 820:825 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 34 theme I 808:808 arg1 activity 810:817 α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity 756:817 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 35 theme β1-2-N-acetylglucosaminyltransferase 771:806 arg1 TbGnTI 820:825 TbGnTI 820:825 TbGnTI 820:825 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 35 theme β1-2-N-acetylglucosaminyltransferase 771:806 arg1 activity 810:817 α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity 756:817 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 5 36 theme protein 902:908 arg1 N-glycans 910:918 significantly modified protein N-glycans 879:918 significantly modified protein N-glycans 879:918 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 6 37 theme TbGnTI 1239:1244 arg1 action 1246:1251 prior TbGnTI action 1233:1251 prior TbGnTI action 1233:1251 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 1 38 theme paucimannose 229:240 arg1 structures 242:251 oligomannose and paucimannose structures 212:251 structures 242:251 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 6 39 theme complex 1077:1083 arg1 biosynthesis 1056:1067 biosynthesis 1056:1067 biosynthesis of both complex and hybrid N-glycans 1056:1104 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 2 40 theme parasite 467:474 arg1 genome 476:481 the parasite genome 463:481 the parasite genome 463:481 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 4 41 theme resident 736:743 arg1 it 707:708 it 707:708 it 707:708 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 41 theme resident 736:743 arg1 UDP-GlcNAc 745:754 a Golgi apparatus resident UDP-GlcNAc 718:754 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 7 42 theme functional 1262:1271 arg1 assay 1282:1286 a functional in vitro assay 1260:1286 a functional in vitro assay 1260:1286 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 7 43 theme metazoan 1434:1441 arg1 GnTIs 1443:1447 metazoan GnTIs 1434:1447 metazoan GnTIs 1434:1447 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 5 44 theme null 857:860 arg1 mutant 862:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant 829:867 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 0 45 theme divergent 59:67 arg1 TbGnTI 104:109 TbGnTI 104:109 TbGnTI 104:109 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 0 45 theme divergent 59:67 arg1 I 101:101 a highly divergent N-acetylglucosaminyltransferase I 50:101 a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) 50:110 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 8 46 theme Sequence 1450:1457 arg1 alignment 1459:1467 Sequence alignment 1450:1467 Sequence alignment 1450:1467 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 7 47 theme biantennary 1335:1345 arg1 Man3GlcNAc2 1347:1357 biantennary Man3GlcNAc2 1335:1357 biantennary Man3GlcNAc2 1335:1357 Using a functional in vitro assay, we showed that TbGnTI transfers UDP-GlcNAc to biantennary Man3GlcNAc2, but not to triantennary Man5GlcNAc2, which is the preferred substrate for metazoan GnTIs. 24550396 6 48 theme prior 1233:1237 arg1 action 1246:1251 prior TbGnTI action 1233:1251 prior TbGnTI action 1233:1251 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 3 49 theme glycosyltransferases 557:576 arg1 family 518:523 a family 516:523 a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family 516:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 5 50 theme normal 924:929 arg1 growth 931:936 normal growth 924:936 normal growth in vitro 924:945 The bloodstream-form TbGT11 null mutant exhibited significantly modified protein N-glycans but normal growth in vitro and infectivity to rodents. 24550396 6 51 dep T. 1107:1108 arg1 brucei 1110:1115 brucei 1110:1115 brucei 1110:1115 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 2 52 located found 454:458 arg2 homologues 342:351 no obvious homologues 331:351 no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 331:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 2 52 located found 454:458 arg1 genome 476:481 the parasite genome 463:481 the parasite genome 463:481 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 2 53 theme β1-4-galactosyltransferase 362:387 arg1 genes 438:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 8 54 theme metazoan 1527:1534 arg1 family 1541:1546 the metazoan GnTI family 1523:1546 the metazoan GnTI family 1523:1546 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 4 55 theme α3-D-mannoside 756:769 arg1 TbGnTI 820:825 TbGnTI 820:825 TbGnTI 820:825 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 55 theme α3-D-mannoside 756:769 arg1 activity 810:817 α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity 756:817 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 2 56 theme known 356:360 arg1 genes 438:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 8 57 attach removed 1510:1516 arg2 enzyme 1496:1501 the T. brucei enzyme 1482:1501 the T. brucei enzyme 1482:1501 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 8 57 attach removed 1510:1516 arg1 family 1541:1546 the metazoan GnTI family 1523:1546 the metazoan GnTI family 1523:1546 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 6 58 theme atypical 1147:1154 arg1 glycans 1171:1177 atypical "pseudohybrid" glycans 1147:1177 atypical "pseudohybrid" glycans 1147:1177 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 4 59 dep UDP-GlcNAc 745:754 arg1 TbGnTI 820:825 TbGnTI 820:825 TbGnTI 820:825 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 4 59 dep UDP-GlcNAc 745:754 arg1 activity 810:817 α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity 756:817 a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI) 718:826 Here we characterize one of these genes, TbGT11, and show that it encodes a Golgi apparatus resident UDP-GlcNAc:α3-D-mannoside β1-2-N-acetylglucosaminyltransferase I activity (TbGnTI). 24550396 1 60 theme diverse 166:172 arg1 N-glycans 188:196 N-glycans 188:196 N-glycans 188:196 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 1 60 theme diverse 166:172 arg1 repertoire 174:183 a diverse repertoire 164:183 a diverse repertoire 164:183 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 3 61 theme putative 528:535 arg1 glycosyltransferases 557:576 putative UDP-sugar-dependent glycosyltransferases 528:576 putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family 528:641 However, we previously reported a family of putative UDP-sugar-dependent glycosyltransferases with similarity to the mammalian β1-3-glycosyltransferase family. 24550396 0 62 theme I 101:101 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 0 62 theme I 101:101 arg1 characterization 30:45 functional characterization 19:45 functional characterization 19:45 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 8 63 theme brucei 1489:1494 arg1 enzyme 1496:1501 the T. brucei enzyme 1482:1501 the T. brucei enzyme 1482:1501 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 8 64 theme GnTI 1536:1539 arg1 family 1541:1546 the metazoan GnTI family 1523:1546 the metazoan GnTI family 1523:1546 Sequence alignment reveals that the T. brucei enzyme is far removed from the metazoan GnTI family and suggests that the parasite has adapted the β3-glycosyltransferase family to catalyze β1-2 linkages. 24550396 2 65 theme genes 438:442 arg1 homologues 342:351 no obvious homologues 331:351 no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 331:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 1 66 theme large 270:274 arg1 N-glycans 284:292 exceptionally large complex N-glycans 256:292 exceptionally large complex N-glycans 256:292 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 24550396 0 67 theme N-acetylglucosaminyltransferase 69:99 arg1 TbGnTI 104:109 TbGnTI 104:109 TbGnTI 104:109 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 0 67 theme N-acetylglucosaminyltransferase 69:99 arg1 I 101:101 a highly divergent N-acetylglucosaminyltransferase I 50:101 a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) 50:110 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 0 68 from characterization 30:45 arg1 brucei 127:132 Trypanosoma brucei 115:132 Trypanosoma brucei 115:132 Identification and functional characterization of a highly divergent N-acetylglucosaminyltransferase I (TbGnTI) in Trypanosoma brucei. 24550396 6 69 theme null 1124:1127 arg1 mutants 1129:1135 T. brucei TbGT11 null mutants 1107:1135 T. brucei TbGT11 null mutants 1107:1135 In contrast to multicellular organisms, where the GnTI reaction is essential for biosynthesis of both complex and hybrid N-glycans, T. brucei TbGT11 null mutants expressed atypical "pseudohybrid" glycans, indicating that TbGnTII activity is not dependent on prior TbGnTI action. 24550396 2 70 theme β1-6-N-acetylglucosaminyltransferase 401:436 arg1 genes 438:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes 356:442 Despite the presence of the latter, no obvious homologues of known β1-4-galactosyltransferase or β1-2- or β1-6-N-acetylglucosaminyltransferase genes have been found in the parasite genome. 24550396 1 71 theme complex 276:282 arg1 N-glycans 284:292 exceptionally large complex N-glycans 256:292 exceptionally large complex N-glycans 256:292 Trypanosoma brucei expresses a diverse repertoire of N-glycans, ranging from oligomannose and paucimannose structures to exceptionally large complex N-glycans. 27474286 0 0 theme specific 109:116 arg1 enrichment 118:127 the specific enrichment 105:127 the specific enrichment of N-linked glycans with simultaneous exclusion of proteins 105:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 7 1 theme glycans 1153:1159 arg1 identification 1135:1148 the identification 1131:1148 the identification of glycans and proteins with other post-translational modifications in biological environment 1131:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 7 2 with glycans 1153:1159 arg1 modifications 1204:1216 other post-translational modifications 1179:1216 other post-translational modifications in biological environment 1179:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 4 3 theme magnetic 627:634 arg1 strategy 668:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 4 4 theme 0.5μg/μL 765:772 arg1 concentration 748:760 a concentration 746:760 a concentration of 0.5μg/μL 746:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 6 5 theme BSA 927:929 arg1 protein 931:937 BSA protein 927:937 BSA protein 927:937 Its selectivity towards BSA protein is 1:200 (molar ratio), showing excellent size-exclusion effect. 27474286 7 6 theme proteins 1165:1172 arg1 identification 1135:1148 the identification 1131:1148 the identification of glycans and proteins with other post-translational modifications in biological environment 1131:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 3 7 theme N-linked 591:598 arg1 glycans 600:606 N-linked glycans 591:606 N-linked glycans 591:606 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 1 8 theme novel 298:302 arg1 approach 304:311 a novel approach 296:311 a novel approach in proteomics research 296:334 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 7 9 from modifications 1204:1216 arg1 environment 1232:1242 biological environment 1221:1242 biological environment 1221:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 3 10 theme glycans 600:606 arg1 enrichment 577:586 the specific enrichment 564:586 the specific enrichment of N-linked glycans in this study 564:620 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 4 11 theme nanoparticles-incorporated 636:661 arg1 strategy 668:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 0 12 theme glycans 141:147 arg1 enrichment 118:127 the specific enrichment 105:127 the specific enrichment of N-linked glycans with simultaneous exclusion of proteins 105:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 6 13 theme molar 949:953 arg1 1:200 942:946 1:200 942:946 1:200 (molar ratio) 942:960 Its selectivity towards BSA protein is 1:200 (molar ratio), showing excellent size-exclusion effect. 27474286 6 13 theme molar 949:953 arg1 ratio 955:959 molar ratio 949:959 molar ratio 949:959 Its selectivity towards BSA protein is 1:200 (molar ratio), showing excellent size-exclusion effect. 27474286 4 14 theme N-linked 704:711 arg1 glycans 713:719 22 N-linked glycans 701:719 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL 701:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 2 15 theme glycans 426:432 arg1 FSPE 409:412 the FSPE 405:412 the FSPE of N-linked glycans 405:432 Notably, silica gel bound with perfluoroalkyl groups was applied to the FSPE of N-linked glycans. 27474286 0 16 theme N-linked 132:139 arg1 glycans 141:147 N-linked glycans 132:147 N-linked glycans with simultaneous exclusion of proteins 132:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 0 17 with glycans 141:147 arg1 exclusion 167:175 simultaneous exclusion 154:175 simultaneous exclusion of proteins 154:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 4 18 theme N-linked 825:832 arg1 glycans 834:840 16 N-linked glycans 822:840 16 N-linked glycans identified 822:851 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 4 19 from digest 734:739 arg1 glycans 713:719 22 N-linked glycans 701:719 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL 701:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 1 20 theme fluorine-fluorine 210:226 arg1 interactions 228:239 fluorine-fluorine interactions 210:239 fluorine-fluorine interactions 210:239 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 0 21 theme modified 9:16 arg1 silica 38:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 7 22 theme new 1114:1116 arg1 prospect 1118:1125 new prospect 1114:1125 new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment 1114:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 4 23 theme OVA 730:732 arg1 digest 734:739 the OVA digest 726:739 the OVA digest with a concentration of 0.5μg/μL 726:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 7 24 theme serum 1089:1093 arg1 opening 1103:1109 opening 1103:1109 opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment 1103:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 7 24 theme serum 1089:1093 arg1 digest 1095:1100 the human serum digest 1079:1100 the human serum digest 1079:1100 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 1 25 theme interactions 228:239 arg1 advantage 197:205 advantage 197:205 advantage of fluorine-fluorine interactions 197:239 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 0 26 theme Fluorous 0:7 arg1 silica 38:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 7 27 theme post-translational 1185:1202 arg1 modifications 1204:1216 other post-translational modifications 1179:1216 other post-translational modifications in biological environment 1179:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 2 28 theme silica 346:351 arg1 gel 353:355 silica gel 346:355 silica gel bound with perfluoroalkyl groups 346:388 Notably, silica gel bound with perfluoroalkyl groups was applied to the FSPE of N-linked glycans. 27474286 0 29 theme mesoporous 27:36 arg1 silica 38:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 1 30 theme proteomics 316:325 arg1 research 327:334 proteomics research 316:334 proteomics research 316:334 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 0 31 theme simultaneous 154:165 arg1 exclusion 167:175 simultaneous exclusion 154:175 simultaneous exclusion of proteins 154:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 1 32 theme fluorous 242:249 arg1 FSPE 275:278 FSPE 275:278 FSPE 275:278 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 1 32 theme fluorous 242:249 arg1 extraction 263:272 fluorous solid-phase extraction 242:272 fluorous solid-phase extraction (FSPE) 242:279 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 0 33 theme magnetic 18:25 arg1 silica 38:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica 0:43 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 1 34 from approach 304:311 arg1 research 327:334 proteomics research 316:334 proteomics research 316:334 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 7 35 with proteins 1165:1172 arg1 modifications 1204:1216 other post-translational modifications 1179:1216 other post-translational modifications in biological environment 1179:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 4 36 theme 5ng/μL 809:814 arg1 limit 800:804 a detection limit 788:804 a detection limit of 5ng/μL (with 16 N-linked glycans identified) 788:852 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 1 37 theme solid-phase 251:261 arg1 FSPE 275:278 FSPE 275:278 FSPE 275:278 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 1 37 theme solid-phase 251:261 arg1 extraction 263:272 fluorous solid-phase extraction 242:272 fluorous solid-phase extraction (FSPE) 242:279 Taking advantage of fluorine-fluorine interactions, fluorous solid-phase extraction (FSPE) is emerging as a novel approach in proteomics research. 27474286 7 38 theme other 1179:1183 arg1 modifications 1204:1216 other post-translational modifications 1179:1216 other post-translational modifications in biological environment 1179:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 4 39 link N-linked 825:832 arg1 glycans 834:840 16 N-linked glycans 822:840 16 N-linked glycans identified 822:851 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 7 40 theme digest 1095:1100 arg1 analysis 1067:1074 the analysis 1063:1074 the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment 1063:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 3 41 theme coated 480:485 arg1 nanoparticles 496:508 mesoporous silica coated magnetic nanoparticles 462:508 mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups 462:541 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 3 42 theme mesoporous 462:471 arg1 nanoparticles 496:508 mesoporous silica coated magnetic nanoparticles 462:508 mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups 462:541 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 3 43 theme previous 444:451 arg1 studies 453:459 previous studies 444:459 previous studies 444:459 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 4 44 with digest 734:739 arg1 concentration 748:760 a concentration 746:760 a concentration of 0.5μg/μL 746:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 5 45 theme good 870:873 arg1 reproducibility 886:900 good day-to-day reproducibility 870:900 good day-to-day reproducibility 870:900 It also showed good day-to-day reproducibility. 27474286 3 46 link N-linked 591:598 arg1 glycans 600:606 N-linked glycans 591:606 N-linked glycans 591:606 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 3 47 theme magnetic 487:494 arg1 nanoparticles 496:508 mesoporous silica coated magnetic nanoparticles 462:508 mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups 462:541 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 0 48 theme solid-phase 78:88 arg1 extraction 90:99 fluorous solid-phase extraction 69:99 fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins 69:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 4 49 with 5ng/μL 809:814 arg1 glycans 834:840 16 N-linked glycans 822:840 16 N-linked glycans identified 822:851 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 7 50 theme human 1083:1087 arg1 opening 1103:1109 opening 1103:1109 opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment 1103:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 7 50 theme human 1083:1087 arg1 digest 1095:1100 the human serum digest 1079:1100 the human serum digest 1079:1100 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 2 51 link N-linked 417:424 arg1 glycans 426:432 N-linked glycans 417:432 N-linked glycans 417:432 Notably, silica gel bound with perfluoroalkyl groups was applied to the FSPE of N-linked glycans. 27474286 0 52 theme fluorous 69:76 arg1 extraction 90:99 fluorous solid-phase extraction 69:99 fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins 69:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 0 53 theme proteins 180:187 arg1 exclusion 167:175 simultaneous exclusion 154:175 simultaneous exclusion of proteins 154:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 2 54 theme perfluoroalkyl 368:381 arg1 groups 383:388 perfluoroalkyl groups 368:388 perfluoroalkyl groups 368:388 Notably, silica gel bound with perfluoroalkyl groups was applied to the FSPE of N-linked glycans. 27474286 6 55 theme size-exclusion 981:994 arg1 effect 996:1001 excellent size-exclusion effect 971:1001 excellent size-exclusion effect 971:1001 Its selectivity towards BSA protein is 1:200 (molar ratio), showing excellent size-exclusion effect. 27474286 7 56 theme present 1021:1027 arg1 method 1029:1034 the present method 1017:1034 the present method 1017:1034 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 7 56 theme present 1021:1027 arg1 effective 1049:1057 effective 1049:1057 effective 1049:1057 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 4 57 theme detection 790:798 arg1 limit 800:804 a detection limit 788:804 a detection limit of 5ng/μL (with 16 N-linked glycans identified) 788:852 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 5 58 theme day-to-day 875:884 arg1 reproducibility 886:900 good day-to-day reproducibility 870:900 good day-to-day reproducibility 870:900 It also showed good day-to-day reproducibility. 27474286 3 59 theme perfluoroalkyl 521:534 arg1 groups 536:541 perfluoroalkyl groups 521:541 perfluoroalkyl groups 521:541 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 3 60 theme silica 473:478 arg1 nanoparticles 496:508 mesoporous silica coated magnetic nanoparticles 462:508 mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups 462:541 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 0 61 link N-linked 132:139 arg1 glycans 141:147 N-linked glycans 132:147 N-linked glycans with simultaneous exclusion of proteins 132:187 Fluorous modified magnetic mesoporous silica composites-incorporated fluorous solid-phase extraction for the specific enrichment of N-linked glycans with simultaneous exclusion of proteins. 27474286 7 62 theme biological 1221:1230 arg1 environment 1232:1242 biological environment 1221:1242 biological environment 1221:1242 In addition, the present method proved to be effective for the analysis of the human serum digest, opening up new prospect for the identification of glycans and proteins with other post-translational modifications in biological environment. 27474286 2 63 theme N-linked 417:424 arg1 glycans 426:432 N-linked glycans 417:432 N-linked glycans 417:432 Notably, silica gel bound with perfluoroalkyl groups was applied to the FSPE of N-linked glycans. 27474286 4 64 theme FSPE 663:666 arg1 strategy 668:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy 623:675 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 6 65 theme excellent 971:979 arg1 effect 996:1001 excellent size-exclusion effect 971:1001 excellent size-exclusion effect 971:1001 Its selectivity towards BSA protein is 1:200 (molar ratio), showing excellent size-exclusion effect. 27474286 4 66 link N-linked 704:711 arg1 glycans 713:719 22 N-linked glycans 701:719 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL 701:772 The magnetic nanoparticles-incorporated FSPE strategy successfully identified 22 N-linked glycans from the OVA digest with a concentration of 0.5μg/μL, and achieved a detection limit of 5ng/μL (with 16 N-linked glycans identified). 27474286 3 67 theme specific 568:575 arg1 enrichment 577:586 the specific enrichment 564:586 the specific enrichment of N-linked glycans in this study 564:620 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 27474286 3 68 from enrichment 577:586 arg1 study 616:620 this study 611:620 this study 611:620 Based on previous studies, mesoporous silica coated magnetic nanoparticles bound with perfluoroalkyl groups were synthesized for the specific enrichment of N-linked glycans in this study. 25175804 0 0 theme Leuconostoc 115:125 arg1 mesenteroides 127:139 Leuconostoc mesenteroides dextransucrase 115:154 Leuconostoc mesenteroides dextransucrase 115:154 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 6 1 theme functional 1404:1413 arg1 applications 1388:1399 potential applications 1378:1399 potential applications 1378:1399 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 1 theme functional 1404:1413 arg1 ingredients 1420:1430 functional food ingredients 1404:1430 functional food ingredients 1404:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 5 2 attach linked 1205:1210 arg2 they 1196:1199 they 1196:1199 they 1196:1199 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 5 2 attach linked 1205:1210 arg3 bonds 1228:1232 only α-(1→6) bonds 1215:1232 only α-(1→6) bonds 1215:1232 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 4 3 theme oligosaccharide 872:886 arg1 content 845:851 the content 841:851 the content of each individual oligosaccharide 841:886 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 5 4 theme produced 1087:1094 arg1 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides (IMOs) 1155:1186 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 5 4 theme produced 1087:1094 arg1 oligosaccharides 1117:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 1 5 from mesenteroides 405:417 arg1 dextransucrase 373:386 a dextransucrase 371:386 a dextransucrase from Leuconostoc mesenteroides B-512F 371:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 2 6 theme oligosaccharide 437:451 arg1 yield 453:457 The total oligosaccharide yield 427:457 The total oligosaccharide yield obtained under optimal conditions 427:491 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 2 6 theme oligosaccharide 437:451 arg1 % 502:502 41-42% 497:502 41-42% 497:502 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 3 7 theme isomaltulose 787:798 arg1 isomaltulose 787:798 isomaltulose 787:798 isomaltulose 787:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 7 theme isomaltulose 787:798 arg1 residue 776:782 the nonreducing glucose residue 752:782 the nonreducing glucose residue of isomaltulose 752:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 2 8 theme isomaltulose 553:564 arg1 amount 543:548 the initial amount 531:548 the initial amount of isomaltulose 531:564 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 2 8 theme isomaltulose 553:564 arg1 isomaltulose 553:564 isomaltulose 553:564 isomaltulose 553:564 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 4 9 theme individual 861:870 arg1 oligosaccharide 872:886 each individual oligosaccharide 856:886 each individual oligosaccharide 856:886 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 4 10 theme content 845:851 arg1 progression 826:836 the progression 822:836 the progression of the content of each individual oligosaccharide 822:886 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 0 11 theme mesenteroides 127:139 arg1 reaction 103:110 transglucosylation reaction 84:110 transglucosylation reaction of Leuconostoc mesenteroides dextransucrase 84:154 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 4 12 theme further 992:998 arg1 transglucosylation 1000:1017 further transglucosylation 992:1017 further transglucosylation to yield oligosaccharides of a higher degree of polymerization 992:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 6 13 theme bioactive 1341:1349 arg1 properties 1351:1360 appealing bioactive properties 1331:1360 appealing bioactive properties that could find potential applications as functional food ingredients 1331:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 2 14 theme reaction 584:591 arg1 h 579:579 24-48 h 573:579 24-48 h of reaction 573:591 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 6 15 theme food 1415:1418 arg1 applications 1388:1399 potential applications 1378:1399 potential applications 1378:1399 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 15 theme food 1415:1418 arg1 ingredients 1420:1430 functional food ingredients 1404:1430 functional food ingredients 1404:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 3 16 theme magnetic 602:609 arg1 NMR 622:624 NMR 622:624 NMR 622:624 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 16 theme magnetic 602:609 arg1 resonance 611:619 Nuclear magnetic resonance 594:619 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 1 17 theme homologous 215:224 arg1 series 226:231 a homologous series 213:231 a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 213:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 1 18 link isomaltulose-derived 236:255 arg1 oligosaccharides 257:272 isomaltulose-derived oligosaccharides 236:272 isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 236:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 0 19 dep mesenteroides 127:139 arg1 dextransucrase 141:154 dextransucrase 141:154 Leuconostoc mesenteroides dextransucrase 115:154 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 1 20 theme series 226:231 arg1 synthesis 200:208 the efficient enzymatic synthesis 176:208 the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 176:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 1 21 theme transglucosylation 337:354 arg1 reaction 356:363 the transglucosylation reaction 333:363 the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 333:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 3 22 theme nonreducing 756:766 arg1 isomaltulose 787:798 isomaltulose 787:798 isomaltulose 787:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 22 theme nonreducing 756:766 arg1 residue 776:782 the nonreducing glucose residue 752:782 the nonreducing glucose residue of isomaltulose 752:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 1 23 theme isomaltulose-derived 236:255 arg1 oligosaccharides 257:272 isomaltulose-derived oligosaccharides 236:272 isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 236:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 6 24 theme appealing 1331:1339 arg1 properties 1351:1360 appealing bioactive properties 1331:1360 appealing bioactive properties that could find potential applications as functional food ingredients 1331:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 4 25 theme higher 1050:1055 arg1 degree 1057:1062 a higher degree 1048:1062 a higher degree of polymerization 1048:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 0 26 link isomaltulose-derived 34:53 arg1 oligosaccharides 55:70 isomaltulose-derived oligosaccharides 34:70 isomaltulose-derived oligosaccharides 34:70 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 6 27 theme core 1275:1278 arg1 structure 1280:1288 the core structure 1271:1288 the core structure 1271:1288 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 27 theme core 1275:1278 arg1 isomaltulose 1255:1266 isomaltulose 1255:1266 isomaltulose 1255:1266 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 28 theme potential 1378:1386 arg1 applications 1388:1399 potential applications 1378:1399 potential applications 1378:1399 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 28 theme potential 1378:1386 arg1 ingredients 1420:1430 functional food ingredients 1404:1430 functional food ingredients 1404:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 1 29 theme oligosaccharides 257:272 arg1 series 226:231 a homologous series 213:231 a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 213:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 6 30 theme IMO-like 1297:1304 arg1 structures 1306:1315 these IMO-like structures 1291:1315 these IMO-like structures 1291:1315 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 3 31 theme structural 627:636 arg1 characterization 638:653 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 0 32 theme oligosaccharides 55:70 arg1 characterization 14:29 characterization 14:29 characterization 14:29 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 0 32 theme oligosaccharides 55:70 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis 0:8 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 5 33 theme isomaltulose-derived 1096:1115 arg1 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides (IMOs) 1155:1186 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 5 33 theme isomaltulose-derived 1096:1115 arg1 oligosaccharides 1117:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 1 34 with oligosaccharides 257:272 arg1 degrees 279:285 degrees 279:285 degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 279:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 4 35 theme low 940:942 arg1 weight 954:959 low molecular weight 940:959 low molecular weight 940:959 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 2 36 theme optimal 474:480 arg1 conditions 482:491 optimal conditions 474:491 optimal conditions 474:491 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 0 37 theme isomaltulose-derived 34:53 arg1 oligosaccharides 55:70 isomaltulose-derived oligosaccharides 34:70 isomaltulose-derived oligosaccharides 34:70 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 6 38 contain possess 1323:1329 arg2 properties 1351:1360 appealing bioactive properties 1331:1360 appealing bioactive properties that could find potential applications as functional food ingredients 1331:1430 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 6 38 contain possess 1323:1329 arg1 structures 1306:1315 these IMO-like structures 1291:1315 these IMO-like structures 1291:1315 In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients. 25175804 4 39 theme degree 1057:1062 arg1 oligosaccharides 1028:1043 oligosaccharides 1028:1043 oligosaccharides of a higher degree of polymerization 1028:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 4 40 theme weight 954:959 arg1 products 928:935 oligosaccharide acceptor products 903:935 oligosaccharide acceptor products of low molecular weight 903:959 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 4 40 theme weight 954:959 arg1 acceptors 978:986 acceptors 978:986 acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization 978:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 2 41 theme total 431:435 arg1 yield 453:457 The total oligosaccharide yield 427:457 The total oligosaccharide yield obtained under optimal conditions 427:491 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 2 41 theme total 431:435 arg1 % 502:502 41-42% 497:502 41-42% 497:502 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 2 42 theme initial 535:541 arg1 amount 543:548 the initial amount 531:548 the initial amount of isomaltulose 531:564 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 2 42 theme initial 535:541 arg1 isomaltulose 553:564 isomaltulose 553:564 isomaltulose 553:564 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 1 43 theme Leuconostoc 393:403 arg1 B-512F 419:424 B-512F 419:424 B-512F 419:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 1 43 theme Leuconostoc 393:403 arg1 mesenteroides 405:417 Leuconostoc mesenteroides 393:417 Leuconostoc mesenteroides B-512F 393:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 3 44 theme glucose 768:774 arg1 isomaltulose 787:798 isomaltulose 787:798 isomaltulose 787:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 44 theme glucose 768:774 arg1 residue 776:782 the nonreducing glucose residue 752:782 the nonreducing glucose residue of isomaltulose 752:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 4 45 theme molecular 944:952 arg1 weight 954:959 low molecular weight 940:959 low molecular weight 940:959 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 3 46 theme residue 776:782 arg1 C-6 745:747 the C-6 741:747 the C-6 of the nonreducing glucose residue of isomaltulose 741:798 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 4 47 theme oligosaccharide 903:917 arg1 products 928:935 oligosaccharide acceptor products 903:935 oligosaccharide acceptor products of low molecular weight 903:959 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 4 47 theme oligosaccharide 903:917 arg1 acceptors 978:986 acceptors 978:986 acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization 978:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 1 48 theme polymerization 290:303 arg1 degrees 279:285 degrees 279:285 degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 279:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 3 49 theme resonance 611:619 arg1 characterization 638:653 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 50 theme sucrose 730:736 arg1 moieties 718:725 glucose moieties 710:725 glucose moieties of sucrose 710:736 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 5 51 link isomaltulose-derived 1096:1115 arg1 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides 1155:1179 isomalto-oligosaccharides (IMOs) 1155:1186 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 5 51 link isomaltulose-derived 1096:1115 arg1 oligosaccharides 1117:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides 1083:1132 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 1 52 theme efficient 180:188 arg1 synthesis 200:208 the efficient enzymatic synthesis 176:208 the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 176:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 1 53 dep 9 323:323 arg1 to 320:321 to 320:321 to 320:321 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 3 54 theme glucose 710:716 arg1 moieties 718:725 glucose moieties 710:725 glucose moieties of sucrose 710:736 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 2 55 from % 502:502 arg1 weight 508:513 weight 508:513 weight with respect to the initial amount of isomaltulose 508:564 The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. 25175804 1 56 theme enzymatic 190:198 arg1 synthesis 200:208 the efficient enzymatic synthesis 176:208 the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F 176:424 This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. 25175804 5 57 dep α- 1220:1221 arg1 1→6 1223:1225 1→6 1223:1225 1→6 1223:1225 The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. 25175804 0 58 theme transglucosylation 84:101 arg1 reaction 103:110 transglucosylation reaction 84:110 transglucosylation reaction of Leuconostoc mesenteroides dextransucrase 84:154 Synthesis and characterization of isomaltulose-derived oligosaccharides produced by transglucosylation reaction of Leuconostoc mesenteroides dextransucrase. 25175804 4 59 theme acceptor 919:926 arg1 products 928:935 oligosaccharide acceptor products 903:935 oligosaccharide acceptor products of low molecular weight 903:959 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 4 59 theme acceptor 919:926 arg1 acceptors 978:986 acceptors 978:986 acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization 978:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 25175804 3 60 theme Nuclear 594:600 arg1 NMR 622:624 NMR 622:624 NMR 622:624 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 3 60 theme Nuclear 594:600 arg1 resonance 611:619 Nuclear magnetic resonance 594:619 Nuclear magnetic resonance (NMR) structural characterization 594:653 Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. 25175804 4 61 theme polymerization 1067:1080 arg1 degree 1057:1062 a higher degree 1048:1062 a higher degree of polymerization 1048:1080 Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. 26760037 7 0 theme dedifferentiated 1813:1828 arg1 cells 1862:1866 dedifferentiated than on the healthy, native RPE cells 1813:1866 dedifferentiated than on the healthy, native RPE cells 1813:1866 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 4 1 theme cell 1051:1054 arg1 surface 1056:1062 the RPE cell surface 1043:1062 the RPE cell surface 1043:1062 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 1 2 from failure 340:346 arg1 surgery 370:376 retinal detachment surgery 351:376 retinal detachment surgery 351:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 2 3 theme RPE 628:630 arg1 cells 632:636 human RPE cells 622:636 human RPE cells 622:636 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 5 4 theme tri-antennary 1413:1425 arg1 N-glycans 1440:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 4 5 theme Gal-3 1034:1038 arg1 binding 1023:1029 binding 1023:1029 binding of Gal-3 to the RPE cell surface 1023:1062 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 5 theme Gal-3 1034:1038 arg1 both 1017:1020 both 1017:1020 both 1017:1020 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 5 theme Gal-3 1034:1038 arg1 inhibition 1083:1092 Gal-3-mediated inhibition 1068:1092 Gal-3-mediated inhibition of RPE attachment and spreading 1068:1124 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 1 6 theme epithelial 212:221 arg1 cells 223:227 retinal pigment epithelial cells 196:227 retinal pigment epithelial cells 196:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 2 7 theme cellular 742:749 arg1 events 751:756 PVR-associated cellular events 727:756 PVR-associated cellular events 727:756 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 9 8 theme therapeutic 2143:2153 arg1 option 2155:2160 a therapeutic option 2141:2160 a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR 2141:2254 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 5 9 theme type 1435:1438 arg1 N-glycans 1440:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 6 10 theme CRISPR-Cas9 1519:1529 arg1 editing 1538:1544 CRISPR-Cas9 genome editing 1519:1544 CRISPR-Cas9 genome editing 1519:1544 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing resulted in reduced Gal-3 binding. 26760037 2 11 theme carbohydrate 546:557 arg1 protein 567:573 a carbohydrate binding protein 544:573 a carbohydrate binding protein 544:573 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 11 theme carbohydrate 546:557 arg1 galectin-3 524:533 galectin-3 524:533 galectin-3 (Gal-3) 524:541 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 1 12 theme crucial 234:240 arg1 transition 176:185 Epithelial-to-mesenchymal transition 150:185 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells 150:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 12 theme crucial 234:240 arg1 event 242:246 a crucial event 232:246 a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery 232:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 7 13 theme Gal-3 1637:1641 arg1 binding 1614:1620 binding 1614:1620 binding of recombinant Gal-3 to the RPE cell surface 1614:1665 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 7 13 theme Gal-3 1637:1641 arg1 effects 1682:1688 inhibitory effects 1671:1688 inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 1671:1764 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 8 14 theme recombinant 2008:2018 arg1 Gal-3 2020:2024 recombinant Gal-3 2008:2024 recombinant Gal-3 2008:2024 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 1 15 from event 242:246 arg1 onset 255:259 the onset 251:259 the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery 251:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 8 16 theme affinity 1988:1995 arg1 binding 1997:2003 high affinity binding 1983:2003 high affinity binding of recombinant Gal-3 1983:2024 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 3 17 theme blot 766:769 arg1 analysis 771:778 Lectin blot analysis 759:778 Lectin blot analysis 759:778 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 4 18 dep found 1006:1010 arg1 Using 969:973 Using 969:973 Using inhibitors of glycosylation 969:1001 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 1 19 theme Epithelial-to-mesenchymal 150:174 arg1 transition 176:185 Epithelial-to-mesenchymal transition 150:185 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells 150:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 19 theme Epithelial-to-mesenchymal 150:174 arg1 event 242:246 a crucial event 232:246 a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery 232:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 19 theme Epithelial-to-mesenchymal 150:174 arg1 EMT 188:190 EMT 188:190 EMT 188:190 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 4 20 theme complex 1200:1206 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 3 21 theme RPE 801:803 arg1 cells 805:809 RPE cells 801:809 RPE cells 801:809 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 3 22 theme glycomic 830:837 arg1 shift 839:843 a glycomic shift 828:843 a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans 828:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 0 23 theme Increased 67:75 arg1 β1,6-N-Glycosylation 77:96 Increased β1,6-N-Glycosylation 67:96 Increased β1,6-N-Glycosylation 67:96 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 6 24 theme Gal-3 1566:1570 arg1 binding 1572:1578 reduced Gal-3 binding 1558:1578 reduced Gal-3 binding 1558:1578 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing resulted in reduced Gal-3 binding. 26760037 3 25 theme Thomsen-Friedenreich 869:888 arg1 antigen 890:896 Thomsen-Friedenreich antigen 869:896 Thomsen-Friedenreich antigen 869:896 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 1 26 theme detachment 359:368 arg1 surgery 370:376 retinal detachment surgery 351:376 retinal detachment surgery 351:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 27 theme retinal 196:202 arg1 cells 223:227 retinal pigment epithelial cells 196:227 retinal pigment epithelial cells 196:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 4 28 theme glycans 1242:1248 arg1 sialylation 1227:1237 sialylation 1227:1237 sialylation of glycans 1227:1248 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 28 theme glycans 1242:1248 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 9 29 theme PVR 2252:2254 arg1 development 2237:2247 development 2237:2247 development of PVR 2237:2254 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 2 30 theme PVR-associated 727:740 arg1 events 751:756 PVR-associated cellular events 727:756 PVR-associated cellular events 727:756 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 31 with interaction 507:517 arg1 galectin-3 524:533 galectin-3 524:533 galectin-3 (Gal-3) 524:541 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 31 with interaction 507:517 arg1 Gal-3 536:540 Gal-3 536:540 Gal-3 536:540 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 31 with interaction 507:517 arg1 protein 567:573 a carbohydrate binding protein 544:573 a carbohydrate binding protein 544:573 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 3 32 theme chains 924:929 arg1 abundance 856:864 an abundance 853:864 an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans 853:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 2 33 theme cells 632:636 arg1 attachment 594:603 attachment 594:603 attachment 594:603 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 33 theme cells 632:636 arg1 spreading 609:617 spreading 609:617 spreading 609:617 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 7 34 theme dependent 1730:1738 arg1 attachment 1697:1706 RPE attachment 1693:1706 RPE attachment 1693:1706 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 2 35 theme surface 414:420 arg1 profile 440:446 the cell surface glycan expression profile 405:446 the cell surface glycan expression profile 405:446 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 0 36 theme Galectin-3 130:139 arg1 Binding 141:147 Galectin-3 Binding 130:147 Galectin-3 Binding 130:147 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 9 37 theme pathogenic 2212:2221 arg1 relevance 2223:2231 pathogenic relevance 2212:2231 pathogenic relevance for development of PVR 2212:2254 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 7 38 theme inhibitory 1671:1680 arg1 effects 1682:1688 inhibitory effects 1671:1688 inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 1671:1764 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 2 39 theme cells 464:468 arg1 EMT 453:455 EMT 453:455 EMT of RPE cells 453:468 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 5 40 theme RPE 1476:1478 arg1 cells 1480:1484 RPE cells 1476:1484 RPE cells 1476:1484 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 0 41 theme Epithelial-to-Mesenchymal 0:24 arg1 Transition 26:35 Epithelial-to-Mesenchymal Transition 0:35 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro 0:57 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 3 42 theme branched 949:956 arg1 N-glycans 958:966 complex-type branched N-glycans 936:966 complex-type branched N-glycans 936:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 4 43 theme RPE 1047:1049 arg1 surface 1056:1062 the RPE cell surface 1043:1062 the RPE cell surface 1043:1062 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 5 44 theme N-acetylglucosaminyltransferase 1317:1347 arg1 V 1349:1349 β1,6 N-acetylglucosaminyltransferase V 1312:1349 β1,6 N-acetylglucosaminyltransferase V (Mgat5) 1312:1357 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 5 44 theme N-acetylglucosaminyltransferase 1317:1347 arg1 enzyme 1368:1373 the key enzyme 1360:1373 the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans 1360:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 5 44 theme N-acetylglucosaminyltransferase 1317:1347 arg1 Mgat5 1352:1356 Mgat5 1352:1356 Mgat5 1352:1356 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 3 45 theme complex-type 936:947 arg1 N-glycans 958:966 complex-type branched N-glycans 936:966 complex-type branched N-glycans 936:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 7 46 theme RPE 1650:1652 arg1 surface 1659:1665 the RPE cell surface 1646:1665 the RPE cell surface 1646:1665 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 8 47 theme myofibroblastic 2063:2077 arg1 RPE 2079:2081 myofibroblastic RPE 2063:2081 myofibroblastic RPE 2063:2081 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 2 48 theme expression 429:438 arg1 profile 440:446 the cell surface glycan expression profile 405:446 the cell surface glycan expression profile 405:446 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 0 49 theme Cells 44:48 arg1 Transition 26:35 Epithelial-to-Mesenchymal Transition 0:35 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro 0:57 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 8 50 theme RPE 1920:1922 arg1 cells 1924:1928 RPE cells 1920:1928 RPE cells 1920:1928 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 8 51 theme Gal-3 2020:2024 arg1 binding 1997:2003 high affinity binding 1983:2003 high affinity binding of recombinant Gal-3 1983:2024 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 1 52 theme treatment 330:338 arg1 failure 340:346 treatment failure 330:346 treatment failure in retinal detachment surgery 330:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 9 53 theme RPE 2197:2199 arg1 cells 2201:2205 RPE cells 2197:2205 RPE cells 2197:2205 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 4 54 theme complex-type 1261:1272 arg1 O-glycans 1274:1282 complex-type O-glycans 1261:1282 complex-type O-glycans 1261:1282 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 55 theme attachment 1101:1110 arg1 binding 1023:1029 binding 1023:1029 binding of Gal-3 to the RPE cell surface 1023:1062 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 55 theme attachment 1101:1110 arg1 both 1017:1020 both 1017:1020 both 1017:1020 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 55 theme attachment 1101:1110 arg1 inhibition 1083:1092 Gal-3-mediated inhibition 1068:1092 Gal-3-mediated inhibition of RPE attachment and spreading 1068:1124 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 9 56 theme future 2091:2096 arg1 perspective 2098:2108 a future perspective 2089:2108 a future perspective 2089:2108 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 1 57 theme vitreoretinopathy 278:294 arg1 onset 255:259 the onset 251:259 the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery 251:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 4 58 theme Gal-3-mediated 1068:1081 arg1 both 1017:1020 both 1017:1020 both 1017:1020 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 58 theme Gal-3-mediated 1068:1081 arg1 inhibition 1083:1092 Gal-3-mediated inhibition 1068:1092 Gal-3-mediated inhibition of RPE attachment and spreading 1068:1124 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 9 59 theme recombinant 2110:2120 arg1 Gal-3 2122:2126 recombinant Gal-3 2110:2126 recombinant Gal-3 2110:2126 From a future perspective recombinant Gal-3 may disclose a therapeutic option allowing for selectively targeting RPE cells with pathogenic relevance for development of PVR. 26760037 4 60 gly sialylation 1227:1237 arg1 glycans 1242:1248 glycans 1242:1248 glycans 1242:1248 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 60 gly sialylation 1227:1237 arg1 interaction 1150:1160 the interaction 1146:1160 the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans 1146:1248 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 1 61 theme pigment 204:210 arg1 cells 223:227 retinal pigment epithelial cells 196:227 retinal pigment epithelial cells 196:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 2 62 theme human 622:626 arg1 cells 632:636 human RPE cells 622:636 human RPE cells 622:636 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 5 63 theme complex 1427:1433 arg1 N-glycans 1440:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 6 64 theme genome 1531:1536 arg1 editing 1538:1544 CRISPR-Cas9 genome editing 1519:1544 CRISPR-Cas9 genome editing 1519:1544 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing resulted in reduced Gal-3 binding. 26760037 1 65 theme cells 223:227 arg1 transition 176:185 Epithelial-to-mesenchymal transition 150:185 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells 150:227 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 65 theme cells 223:227 arg1 event 242:246 a crucial event 232:246 a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery 232:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 65 theme cells 223:227 arg1 EMT 188:190 EMT 188:190 EMT 188:190 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 2 66 from alterations 390:400 arg1 profile 440:446 the cell surface glycan expression profile 405:446 the cell surface glycan expression profile 405:446 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 4 67 theme spreading 1116:1124 arg1 binding 1023:1029 binding 1023:1029 binding of Gal-3 to the RPE cell surface 1023:1062 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 67 theme spreading 1116:1124 arg1 both 1017:1020 both 1017:1020 both 1017:1020 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 67 theme spreading 1116:1124 arg1 inhibition 1083:1092 Gal-3-mediated inhibition 1068:1092 Gal-3-mediated inhibition of RPE attachment and spreading 1068:1124 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 3 68 theme Lectin 759:764 arg1 analysis 771:778 Lectin blot analysis 759:778 Lectin blot analysis 759:778 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 5 69 theme N-glycans 1440:1448 arg1 synthesis 1390:1398 the synthesis 1386:1398 the synthesis of tetra- or tri-antennary complex type N-glycans 1386:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 7 70 theme recombinant 1625:1635 arg1 Gal-3 1637:1641 recombinant Gal-3 1625:1641 recombinant Gal-3 1625:1641 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 4 71 theme tetra-antennary 1184:1198 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 8 72 theme cells 1924:1928 arg1 EMT 1913:1915 EMT 1913:1915 EMT of RPE cells in vitro 1913:1937 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 4 73 theme type 1208:1211 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 2 74 theme binding 559:565 arg1 protein 567:573 a carbohydrate binding protein 544:573 a carbohydrate binding protein 544:573 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 2 74 theme binding 559:565 arg1 galectin-3 524:533 galectin-3 524:533 galectin-3 (Gal-3) 524:541 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 7 75 from effects 1682:1688 arg1 spreading 1712:1720 spreading 1712:1720 spreading 1712:1720 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 7 75 from effects 1682:1688 arg1 attachment 1697:1706 RPE attachment 1693:1706 RPE attachment 1693:1706 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 4 76 theme Gal-3 1165:1169 arg1 interaction 1150:1160 the interaction 1146:1160 the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans 1146:1248 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 3 77 theme cells 805:809 arg1 EMT 794:796 EMT 794:796 EMT of RPE cells in vitro 794:818 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 8 78 theme glycomic 1947:1954 arg1 changes 1956:1962 glycomic changes 1947:1962 glycomic changes 1947:1962 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 4 79 theme glycosylation 989:1001 arg1 inhibitors 975:984 inhibitors 975:984 inhibitors of glycosylation 975:1001 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 80 theme tri- 1176:1179 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 0 81 theme Increased 102:110 arg1 Susceptibility 112:125 Increased Susceptibility 102:125 Increased Susceptibility to Galectin-3 Binding 102:147 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 1 82 theme retinal 351:357 arg1 detachment 359:368 retinal detachment 351:368 retinal detachment surgery 351:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 6 83 theme reduced 1558:1564 arg1 binding 1572:1578 reduced Gal-3 binding 1558:1578 reduced Gal-3 binding 1558:1578 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing resulted in reduced Gal-3 binding. 26760037 3 84 theme antigen 890:896 arg1 abundance 856:864 an abundance 853:864 an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans 853:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 7 85 from binding 1614:1620 arg1 spreading 1712:1720 spreading 1712:1720 spreading 1712:1720 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 7 85 from binding 1614:1620 arg1 attachment 1697:1706 RPE attachment 1693:1706 RPE attachment 1693:1706 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 8 86 theme high 1983:1986 arg1 binding 1997:2003 high affinity binding 1983:2003 high affinity binding of recombinant Gal-3 1983:2024 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 7 87 theme RPE 1693:1695 arg1 attachment 1697:1706 RPE attachment 1693:1706 RPE attachment 1693:1706 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 8 88 theme cell 2047:2050 arg1 surface 2052:2058 the cell surface 2043:2058 the cell surface of myofibroblastic RPE 2043:2081 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 7 89 with interaction 1743:1753 arg1 Mgat5 1760:1764 Mgat5 1760:1764 Mgat5 1760:1764 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 2 90 theme cell 409:412 arg1 profile 440:446 the cell surface glycan expression profile 405:446 the cell surface glycan expression profile 405:446 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 3 91 theme poly-N-acetyllactosamine 899:922 arg1 chains 924:929 poly-N-acetyllactosamine chains 899:929 poly-N-acetyllactosamine chains 899:929 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 2 92 theme RPE 460:462 arg1 cells 464:468 RPE cells 460:468 RPE cells 460:468 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 5 93 theme cells 1480:1484 arg1 EMT 1469:1471 EMT 1469:1471 EMT of RPE cells 1469:1484 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 2 94 theme carbohydrate-dependent 653:674 arg1 manner 676:681 carbohydrate-dependent manner 653:681 carbohydrate-dependent manner 653:681 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 5 95 theme β1,6 1312:1315 arg1 V 1349:1349 β1,6 N-acetylglucosaminyltransferase V 1312:1349 β1,6 N-acetylglucosaminyltransferase V (Mgat5) 1312:1357 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 5 95 theme β1,6 1312:1315 arg1 enzyme 1368:1373 the key enzyme 1360:1373 the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans 1360:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 5 95 theme β1,6 1312:1315 arg1 Mgat5 1352:1356 Mgat5 1352:1356 Mgat5 1352:1356 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 0 96 theme RPE 40:42 arg1 Cells 44:48 RPE Cells 40:48 RPE Cells 40:48 Epithelial-to-Mesenchymal Transition of RPE Cells In Vitro Confers Increased β1,6-N-Glycosylation and Increased Susceptibility to Galectin-3 Binding. 26760037 4 97 with interaction 1150:1160 arg1 sialylation 1227:1237 sialylation 1227:1237 sialylation of glycans 1227:1248 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 4 97 with interaction 1150:1160 arg1 N-glycans 1213:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 tri- or tetra-antennary complex type N-glycans 1176:1221 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 3 98 theme N-glycans 958:966 arg1 abundance 856:864 an abundance 853:864 an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans 853:966 Lectin blot analysis revealed that EMT of RPE cells in vitro confers a glycomic shift towards an abundance of Thomsen-Friedenreich antigen, poly-N-acetyllactosamine chains, and complex-type branched N-glycans. 26760037 8 99 theme RPE 2079:2081 arg1 surface 2052:2058 the cell surface 2043:2058 the cell surface of myofibroblastic RPE 2043:2081 Based on these findings we hypothesize that EMT of RPE cells in vitro confers glycomic changes, which account for high affinity binding of recombinant Gal-3, particularly to the cell surface of myofibroblastic RPE. 26760037 1 100 theme common 312:317 arg1 vitreoretinopathy 278:294 proliferative vitreoretinopathy 264:294 proliferative vitreoretinopathy (PVR) 264:300 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 100 theme common 312:317 arg1 reason 319:324 the most common reason 303:324 the most common reason for treatment failure in retinal detachment surgery 303:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 7 101 theme cell 1654:1657 arg1 surface 1659:1665 the RPE cell surface 1646:1665 the RPE cell surface 1646:1665 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 2 102 theme glycan 422:427 arg1 profile 440:446 the cell surface glycan expression profile 405:446 the cell surface glycan expression profile 405:446 We studied alterations in the cell surface glycan expression profile upon EMT of RPE cells and focused on its relevance for the interaction with galectin-3 (Gal-3), a carbohydrate binding protein, which can inhibit attachment and spreading of human RPE cells in a dose- and carbohydrate-dependent manner, and thus bares the potential to counteract PVR-associated cellular events. 26760037 7 103 theme native 1851:1856 arg1 RPE 1858:1860 the healthy, native RPE 1838:1860 RPE 1858:1860 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 5 104 theme tetra- 1403:1408 arg1 N-glycans 1440:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 tetra- or tri-antennary complex type N-glycans 1403:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 6 105 theme Mgat5 1500:1504 arg1 Silencing 1487:1495 Silencing 1487:1495 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing 1487:1544 Silencing of Mgat5 by siRNA and CRISPR-Cas9 genome editing resulted in reduced Gal-3 binding. 26760037 7 106 theme healthy 1842:1848 arg1 RPE 1858:1860 the healthy, native RPE 1838:1860 RPE 1858:1860 We conclude from these data that binding of recombinant Gal-3 to the RPE cell surface and inhibitory effects on RPE attachment and spreading largely dependent on interaction with Mgat5 modified N-glycans, which are more abundant on dedifferentiated than on the healthy, native RPE cells. 26760037 5 107 theme key 1364:1366 arg1 V 1349:1349 β1,6 N-acetylglucosaminyltransferase V 1312:1349 β1,6 N-acetylglucosaminyltransferase V (Mgat5) 1312:1357 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 5 107 theme key 1364:1366 arg1 enzyme 1368:1373 the key enzyme 1360:1373 the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans 1360:1448 Importantly, we found that β1,6 N-acetylglucosaminyltransferase V (Mgat5), the key enzyme catalyzing the synthesis of tetra- or tri-antennary complex type N-glycans, is increased upon EMT of RPE cells. 26760037 4 108 theme RPE 1097:1099 arg1 attachment 1101:1110 RPE attachment 1097:1110 RPE attachment 1097:1110 Using inhibitors of glycosylation we found that both, binding of Gal-3 to the RPE cell surface and Gal-3-mediated inhibition of RPE attachment and spreading, strongly depend on the interaction of Gal-3 with tri- or tetra-antennary complex type N-glycans and sialylation of glycans but not on complex-type O-glycans. 26760037 1 109 theme proliferative 264:276 arg1 PVR 297:299 PVR 297:299 PVR 297:299 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 109 theme proliferative 264:276 arg1 reason 319:324 the most common reason 303:324 the most common reason for treatment failure in retinal detachment surgery 303:376 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26760037 1 109 theme proliferative 264:276 arg1 vitreoretinopathy 278:294 proliferative vitreoretinopathy 264:294 proliferative vitreoretinopathy (PVR) 264:300 Epithelial-to-mesenchymal transition (EMT) of retinal pigment epithelial cells is a crucial event in the onset of proliferative vitreoretinopathy (PVR), the most common reason for treatment failure in retinal detachment surgery. 26108336 5 0 theme optimal 579:585 arg1 temperature 594:604 temperature 594:604 temperature 594:604 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 5 0 theme optimal 579:585 arg1 pH 587:588 optimal pH 579:588 optimal pH 579:588 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 5 0 theme optimal 579:585 arg1 °C 631:632 7.0 and 40 °C 620:632 7.0 and 40 °C 620:632 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 10 1 theme first 1303:1307 arg1 This 1291:1294 This 1291:1294 This 1291:1294 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 10 1 theme first 1303:1307 arg1 report 1309:1314 the first report 1299:1314 the first report of a GH19 chitinase in fungi and in Microsporidia 1299:1364 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 3 2 theme metal 404:408 arg1 chromatography 419:432 metal affinity chromatography 404:432 metal affinity chromatography 404:432 Here, a chitinase from Nosema bombycis (NbchiA) was cloned and purified by metal affinity chromatography and molecular exclusion chromatography. 26108336 7 3 theme chitin 792:797 arg1 GlcNAc 817:822 GlcNAc 817:822 GlcNAc 817:822 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 7 3 theme chitin 792:797 arg1 2-5 825:827 2-5 825:827 2-5 825:827 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 7 3 theme chitin 792:797 arg1 oligosaccharides 799:814 chitin oligosaccharides 792:814 chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography 792:879 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 1 4 theme 1,4 190:192 arg1 linkages 195:202 the β-(1,4) linkages 183:202 the β-(1,4) linkages of chitin 183:212 Chitinases (EC 3.2.1.14), as one kind of glycosyl hydrolase, hydrolyze the β-(1,4) linkages of chitin. 26108336 7 5 theme liquid 859:864 arg1 chromatography 866:879 high-performance liquid chromatography 842:879 high-performance liquid chromatography 842:879 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 6 6 theme ethylene 728:735 arg1 glycol 737:742 ethylene glycol chitin and soluble chitosan 728:770 glycol 737:742 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 3 7 theme exclusion 448:456 arg1 chromatography 458:471 molecular exclusion chromatography 438:471 molecular exclusion chromatography 438:471 Here, a chitinase from Nosema bombycis (NbchiA) was cloned and purified by metal affinity chromatography and molecular exclusion chromatography. 26108336 7 8 theme oligosaccharides 799:814 arg1 degradation 777:787 The degradation 773:787 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography 773:879 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 9 9 theme Glu60 1216:1220 arg1 mutation 1204:1211 mutation 1204:1211 mutation of Glu60 1204:1220 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 10 10 theme chitinase 1326:1334 arg1 This 1291:1294 This 1291:1294 This 1291:1294 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 10 10 theme chitinase 1326:1334 arg1 report 1309:1314 the first report 1299:1314 the first report of a GH19 chitinase in fungi and in Microsporidia 1299:1364 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 4 11 theme class 555:559 arg1 chitinase 564:572 class IV chitinase 555:572 glycoside hydrolase family 19 class IV chitinase 525:572 Sequence analysis indicated that NbchiA belongs to glycoside hydrolase family 19 class IV chitinase. 26108336 0 12 theme Nosema 90:95 arg1 Nb 107:108 Nb 107:108 Nb 107:108 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 0 12 theme Nosema 90:95 arg1 bombycis 97:104 Nosema bombycis 90:104 Nosema bombycis (Nb) 90:109 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 7 13 theme high-performance 842:857 arg1 chromatography 866:879 high-performance liquid chromatography 842:879 high-performance liquid chromatography 842:879 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 3 14 theme molecular 438:446 arg1 chromatography 458:471 molecular exclusion chromatography 438:471 molecular exclusion chromatography 438:471 Here, a chitinase from Nosema bombycis (NbchiA) was cloned and purified by metal affinity chromatography and molecular exclusion chromatography. 26108336 5 15 dep pH 587:588 arg1 The 575:577 The 575:577 The 575:577 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 10 16 theme GH19 1321:1324 arg1 chitinase 1326:1334 a GH19 chitinase 1319:1334 a GH19 chitinase 1319:1334 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 6 17 theme soluble 701:707 arg1 chitosan 763:770 soluble chitosan 755:770 soluble chitosan 755:770 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 6 17 theme soluble 701:707 arg1 substrates 709:718 soluble substrates 701:718 soluble substrates such as ethylene glycol chitin and soluble chitosan 701:770 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 6 17 theme soluble 701:707 arg1 glycol 737:742 ethylene glycol chitin and soluble chitosan 728:770 glycol 737:742 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 4 18 theme Sequence 474:481 arg1 analysis 483:490 Sequence analysis 474:490 Sequence analysis 474:490 Sequence analysis indicated that NbchiA belongs to glycoside hydrolase family 19 class IV chitinase. 26108336 8 19 dep alignment 1038:1046 arg1 the 991:993 the 991:993 the 991:993 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 8 19 dep alignment 1038:1046 arg1 basis 995:999 basis 995:999 basis 995:999 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 2 20 theme sequence 232:239 arg1 similarity 241:250 the sequence similarity 228:250 the sequence similarity 228:250 According to the sequence similarity, chitinases can be divided into glycoside hydrolase family 18 and family 19. 26108336 0 21 theme First 24:28 arg1 Family 56:61 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 1 22 theme chitin 207:212 arg1 linkages 195:202 the β-(1,4) linkages 183:202 the β-(1,4) linkages of chitin 183:212 Chitinases (EC 3.2.1.14), as one kind of glycosyl hydrolase, hydrolyze the β-(1,4) linkages of chitin. 26108336 9 23 theme site-directed 1116:1128 arg1 analysis 1139:1146 The site-directed mutation analysis 1112:1146 The site-directed mutation analysis 1112:1146 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 8 24 theme key 1088:1090 arg1 Glu51 1049:1053 Glu51 1049:1053 Glu51 1049:1053 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 8 24 theme key 1088:1090 arg1 Glu60 1059:1063 Glu60 1059:1063 Glu60 1059:1063 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 8 24 theme key 1088:1090 arg1 residues 1102:1109 the key catalytic residues 1084:1109 the key catalytic residues 1084:1109 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 9 25 theme mutation 1130:1137 arg1 analysis 1139:1146 The site-directed mutation analysis 1112:1146 The site-directed mutation analysis 1112:1146 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 6 26 dep glycol 737:742 arg1 chitin 744:749 chitin 744:749 chitin 744:749 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 2 27 theme hydrolase 294:302 arg1 family 304:309 glycoside hydrolase family 18 284:312 glycoside hydrolase family 18 284:312 According to the sequence similarity, chitinases can be divided into glycoside hydrolase family 18 and family 19. 26108336 0 28 theme Glycosyl 37:44 arg1 Family 56:61 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 7 29 theme GlcNAc 974:979 arg1 end 966:968 the reducing end 953:968 the reducing end of (GlcNAc)(3-5) 953:985 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 0 30 dep Family 56:61 arg1 NbchiA 77:82 NbchiA 77:82 NbchiA 77:82 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 0 30 dep Family 56:61 arg1 Chitinase 66:74 Chitinase 66:74 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 1 31 theme glycosyl 153:160 arg1 hydrolase 162:170 glycosyl hydrolase 153:170 glycosyl hydrolase 153:170 Chitinases (EC 3.2.1.14), as one kind of glycosyl hydrolase, hydrolyze the β-(1,4) linkages of chitin. 26108336 0 32 theme Fungal 30:35 arg1 Family 56:61 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 8 33 theme catalytic 1092:1100 arg1 Glu51 1049:1053 Glu51 1049:1053 Glu51 1049:1053 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 8 33 theme catalytic 1092:1100 arg1 Glu60 1059:1063 Glu60 1059:1063 Glu60 1059:1063 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 8 33 theme catalytic 1092:1100 arg1 residues 1102:1109 the key catalytic residues 1084:1109 the key catalytic residues 1084:1109 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 3 34 from bombycis 359:366 arg1 chitinase 337:345 a chitinase 335:345 a chitinase from Nosema bombycis (NbchiA) 335:375 Here, a chitinase from Nosema bombycis (NbchiA) was cloned and purified by metal affinity chromatography and molecular exclusion chromatography. 26108336 1 35 theme hydrolase 162:170 arg1 kind 145:148 one kind 141:148 one kind of glycosyl hydrolase 141:170 Chitinases (EC 3.2.1.14), as one kind of glycosyl hydrolase, hydrolyze the β-(1,4) linkages of chitin. 26108336 0 36 theme Family 56:61 arg1 Characterization 0:15 Characterization 0:15 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 0:110 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 9 37 theme enzymatic 1271:1279 arg1 activity 1281:1288 the enzymatic activity 1267:1288 the enzymatic activity 1267:1288 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 9 38 theme Glu51 1243:1247 arg1 mutation 1231:1238 mutation 1231:1238 mutation of Glu51 1231:1247 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 7 39 theme reducing 957:964 arg1 end 966:968 the reducing end 953:968 the reducing end of (GlcNAc)(3-5) 953:985 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 0 40 theme Hydrolase 46:54 arg1 Family 56:61 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) 20:83 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 0 41 from bombycis 97:104 arg1 Characterization 0:15 Characterization 0:15 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 0:110 Characterization of the First Fungal Glycosyl Hydrolase Family 19 Chitinase (NbchiA) from Nosema bombycis (Nb). 26108336 6 42 theme soluble 755:761 arg1 chitosan 763:770 soluble chitosan 755:770 soluble chitosan 755:770 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 7 43 theme glycosidic 929:938 arg1 linkage 940:946 the second glycosidic linkage 918:946 the second glycosidic linkage from the reducing end of (GlcNAc)(3-5) 918:985 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 5 44 theme NbchiA 609:614 arg1 temperature 594:604 temperature 594:604 temperature 594:604 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 5 44 theme NbchiA 609:614 arg1 pH 587:588 optimal pH 579:588 optimal pH 579:588 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 5 44 theme NbchiA 609:614 arg1 °C 631:632 7.0 and 40 °C 620:632 7.0 and 40 °C 620:632 The optimal pH and temperature of NbchiA are 7.0 and 40 °C, respectively. 26108336 4 45 dep family 545:550 arg1 chitinase 564:572 class IV chitinase 555:572 glycoside hydrolase family 19 class IV chitinase 525:572 Sequence analysis indicated that NbchiA belongs to glycoside hydrolase family 19 class IV chitinase. 26108336 6 46 theme purified 654:661 arg1 chitinase 663:671 This purified chitinase 649:671 This purified chitinase 649:671 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 4 47 theme hydrolase 535:543 arg1 family 545:550 glycoside hydrolase family 19 class IV chitinase 525:572 glycoside hydrolase family 19 class IV chitinase 525:572 Sequence analysis indicated that NbchiA belongs to glycoside hydrolase family 19 class IV chitinase. 26108336 8 48 theme structure-based 1004:1018 arg1 alignment 1038:1046 structure-based multiple-sequence alignment 1004:1046 structure-based multiple-sequence alignment 1004:1046 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 6 49 theme high 680:683 arg1 activity 685:692 high activity 680:692 high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan 680:770 This purified chitinase showed high activity toward soluble substrates such as ethylene glycol chitin and soluble chitosan. 26108336 8 50 theme multiple-sequence 1020:1036 arg1 alignment 1038:1046 structure-based multiple-sequence alignment 1004:1046 structure-based multiple-sequence alignment 1004:1046 On the basis of structure-based multiple-sequence alignment, Glu51 and Glu60 are believed to be the key catalytic residues. 26108336 10 51 from report 1309:1314 arg1 fungi 1339:1343 fungi 1339:1343 fungi 1339:1343 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 10 51 from report 1309:1314 arg1 Microsporidia 1352:1364 Microsporidia 1352:1364 Microsporidia 1352:1364 This is the first report of a GH19 chitinase in fungi and in Microsporidia. 26108336 4 52 theme glycoside 525:533 arg1 family 545:550 glycoside hydrolase family 19 class IV chitinase 525:572 glycoside hydrolase family 19 class IV chitinase 525:572 Sequence analysis indicated that NbchiA belongs to glycoside hydrolase family 19 class IV chitinase. 26108336 7 53 theme second 922:927 arg1 linkage 940:946 the second glycosidic linkage 918:946 the second glycosidic linkage from the reducing end of (GlcNAc)(3-5) 918:985 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 3 54 theme affinity 410:417 arg1 chromatography 419:432 metal affinity chromatography 404:432 metal affinity chromatography 404:432 Here, a chitinase from Nosema bombycis (NbchiA) was cloned and purified by metal affinity chromatography and molecular exclusion chromatography. 26108336 9 55 theme enzymatic 1166:1174 arg1 activity 1176:1183 the enzymatic activity 1162:1183 the enzymatic activity 1162:1183 The site-directed mutation analysis revealed that the enzymatic activity was decreased upon mutation of Glu60, whereas mutation of Glu51 totally abolished the enzymatic activity. 26108336 7 56 from end 966:968 arg1 linkage 940:946 the second glycosidic linkage 918:946 the second glycosidic linkage from the reducing end of (GlcNAc)(3-5) 918:985 The degradation of chitin oligosaccharides (GlcNAc)(2-5) detected by high-performance liquid chromatography showed that NbchiA hydrolyzed mainly the second glycosidic linkage from the reducing end of (GlcNAc)(3-5). 26108336 2 57 theme glycoside 284:292 arg1 family 304:309 glycoside hydrolase family 18 284:312 glycoside hydrolase family 18 284:312 According to the sequence similarity, chitinases can be divided into glycoside hydrolase family 18 and family 19. 26108336 1 58 theme β- 187:188 arg1 linkages 195:202 the β-(1,4) linkages 183:202 the β-(1,4) linkages of chitin 183:212 Chitinases (EC 3.2.1.14), as one kind of glycosyl hydrolase, hydrolyze the β-(1,4) linkages of chitin. 26100685 5 0 theme preventive 990:999 arg1 applications 1001:1012 the possible diagnostic and preventive applications 962:1012 the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes 962:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 1 1 theme much 202:205 arg1 attention 207:215 much attention 202:215 much attention 202:215 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 2 2 from effect 233:238 arg1 aggregation/fibrillation 342:365 the aggregation/fibrillation 338:365 the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis 338:445 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 4 3 theme BSA 845:847 arg1 amyloidogenesis 826:840 amyloidogenesis 826:840 amyloidogenesis 826:840 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 3 theme BSA 845:847 arg1 aggregation 810:820 aggregation 810:820 aggregation 810:820 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 4 used utilized 753:760 arg2 spectropolarimetry 729:746 spectropolarimetry 729:746 spectropolarimetry 729:746 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 4 used utilized 753:760 arg2 spectrofluorimetry 706:723 spectrofluorimetry 706:723 spectrofluorimetry 706:723 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 5 from effect 777:782 arg1 amyloidogenesis 826:840 amyloidogenesis 826:840 amyloidogenesis 826:840 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 5 from effect 777:782 arg1 aggregation 810:820 aggregation 810:820 aggregation 810:820 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 3 6 dep spectroscopy 515:526 arg1 methods 674:680 methods 674:680 methods 674:680 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 5 7 theme BSA 872:874 arg1 amyloidogenesis 876:890 The BSA amyloidogenesis 868:890 The BSA amyloidogenesis 868:890 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 4 8 theme β-CD/Fe3O4 787:796 arg1 MNPs 798:801 β-CD/Fe3O4 MNPs 787:801 β-CD/Fe3O4 MNPs 787:801 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 4 9 dep aggregation 810:820 arg1 the 806:808 the 806:808 the 806:808 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 5 10 theme degenerative 1026:1037 arg1 effects 1039:1045 the degenerative effects 1022:1045 the degenerative effects of protein glycation/fibrillation under diabetes 1022:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 5 11 theme possible 966:973 arg1 applications 1001:1012 the possible diagnostic and preventive applications 962:1012 the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes 962:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 1 12 theme glycation 118:126 arg1 aggregation 144:154 glycation induced protein aggregation 118:154 glycation induced protein aggregation 118:154 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 2 13 theme diabetic 403:410 arg1 condition 412:420 diabetic condition 403:420 diabetic condition known as amyloidogenesis 403:445 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 0 14 theme Albumin 20:26 arg1 Glycation/Fibrillation 28:49 Serum Albumin Glycation/Fibrillation 14:49 Serum Albumin Glycation/Fibrillation 14:49 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 5 15 theme protein 1050:1056 arg1 glycation/fibrillation 1058:1079 protein glycation/fibrillation 1050:1079 protein glycation/fibrillation under diabetes 1050:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 1 16 theme induced 128:134 arg1 aggregation 144:154 glycation induced protein aggregation 118:154 glycation induced protein aggregation 118:154 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 0 17 theme Serum 14:18 arg1 Glycation/Fibrillation 28:49 Serum Albumin Glycation/Fibrillation 14:49 Serum Albumin Glycation/Fibrillation 14:49 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 5 18 theme diagnostic 975:984 arg1 applications 1001:1012 the possible diagnostic and preventive applications 962:1012 the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes 962:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 3 19 theme sample 648:653 arg1 VSM 669:671 VSM 669:671 VSM 669:671 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 19 theme sample 648:653 arg1 magnetometer 655:666 vibrating sample magnetometer 638:666 vibrating sample magnetometer (VSM) 638:672 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 1 20 theme protein 136:142 arg1 aggregation 144:154 glycation induced protein aggregation 118:154 glycation induced protein aggregation 118:154 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 3 21 theme transmission 595:606 arg1 TEM 629:631 TEM 629:631 TEM 629:631 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 21 theme transmission 595:606 arg1 microscopy 617:626 transmission electron microscopy 595:626 transmission electron microscopy (TEM) 595:632 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 22 theme β-CD 463:466 arg1 conjugation 468:478 the β-CD conjugation 459:478 the β-CD conjugation on MNP 459:485 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 0 23 theme Glycation/Fibrillation 28:49 arg1 Prevention 0:9 Prevention 0:9 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 0:106 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 2 24 theme nanoparticles 278:290 arg1 effect 233:238 the effect 229:238 the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis 229:445 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 0 25 theme Functionalized 69:82 arg1 Nanoparticles 93:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 2 26 with Fe3O4 295:299 arg1 β-CD 329:332 β-CD 329:332 β-CD 329:332 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 26 with Fe3O4 295:299 arg1 β-cyclodextrin 313:326 β-cyclodextrin 313:326 β-cyclodextrin (β-CD) 313:333 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 3 27 theme vibrating 638:646 arg1 VSM 669:671 VSM 669:671 VSM 669:671 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 27 theme vibrating 638:646 arg1 magnetometer 655:666 vibrating sample magnetometer 638:666 vibrating sample magnetometer (VSM) 638:672 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 2 28 theme core-shell 267:276 arg1 nanoparticles 278:290 functionalized magnetic core-shell nanoparticles 243:290 functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) 243:333 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 29 theme albumin 383:389 arg1 aggregation/fibrillation 342:365 the aggregation/fibrillation 338:365 the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis 338:445 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 0 30 theme β-Cyclodextrin 54:67 arg1 Nanoparticles 93:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 3 31 theme infrared 506:513 arg1 FTIR 529:532 FTIR 529:532 FTIR 529:532 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 31 theme infrared 506:513 arg1 spectroscopy 515:526 infrared spectroscopy 506:526 infrared spectroscopy (FTIR) 506:533 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 2 32 theme functionalized 243:256 arg1 nanoparticles 278:290 functionalized magnetic core-shell nanoparticles 243:290 functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) 243:333 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 33 theme magnetic 258:265 arg1 nanoparticles 278:290 functionalized magnetic core-shell nanoparticles 243:290 functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) 243:333 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 34 theme serum 377:381 arg1 BSA 392:394 BSA 392:394 BSA 392:394 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 34 theme serum 377:381 arg1 albumin 383:389 bovine serum albumin 370:389 bovine serum albumin (BSA) 370:395 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 5 35 theme glycation/fibrillation 1058:1079 arg1 effects 1039:1045 the degenerative effects 1022:1045 the degenerative effects of protein glycation/fibrillation under diabetes 1022:1094 The BSA amyloidogenesis was significantly inhibited by interfering β-CD-MNPs that may present the possible diagnostic and preventive applications against the degenerative effects of protein glycation/fibrillation under diabetes. 26100685 3 36 theme electron 608:615 arg1 TEM 629:631 TEM 629:631 TEM 629:631 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 36 theme electron 608:615 arg1 microscopy 617:626 transmission electron microscopy 595:626 transmission electron microscopy (TEM) 595:632 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 1 37 theme related 160:166 arg1 strategies 177:186 related opposing strategies 160:186 related opposing strategies 160:186 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 3 38 theme thermogravimetric 561:577 arg1 analysis 579:586 thermogravimetric analysis 561:586 thermogravimetric analysis (TGA) 561:592 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 38 theme thermogravimetric 561:577 arg1 TGA 589:591 TGA 589:591 TGA 589:591 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 4 39 theme MNPs 798:801 arg1 effect 777:782 the effect 773:782 the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation 773:865 Moreover, spectrofluorimetry and spectropolarimetry were utilized to analyze the effect of β-CD/Fe3O4 MNPs on the aggregation and amyloidogenesis of BSA through glycation. 26100685 1 40 theme opposing 168:175 arg1 strategies 177:186 related opposing strategies 160:186 related opposing strategies 160:186 Nowadays, glycation induced protein aggregation and related opposing strategies have received much attention. 26100685 2 41 theme bovine 370:375 arg1 BSA 392:394 BSA 392:394 BSA 392:394 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 2 41 theme bovine 370:375 arg1 albumin 383:389 bovine serum albumin 370:389 bovine serum albumin (BSA) 370:395 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 3 42 dep Fourier 488:494 arg1 transform 496:504 transform 496:504 transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods 496:680 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 43 theme X-ray 536:540 arg1 XRD 555:557 XRD 555:557 XRD 555:557 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 3 43 theme X-ray 536:540 arg1 diffraction 542:552 X-ray diffraction 536:552 X-ray diffraction (XRD) 536:558 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 26100685 0 44 theme Magnetic 84:91 arg1 Nanoparticles 93:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 β-Cyclodextrin Functionalized Magnetic Nanoparticles 54:105 Prevention of Serum Albumin Glycation/Fibrillation by β-Cyclodextrin Functionalized Magnetic Nanoparticles. 26100685 2 45 theme Fe3O4 295:299 arg1 nanoparticles 278:290 functionalized magnetic core-shell nanoparticles 243:290 functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) 243:333 We present the effect of functionalized magnetic core-shell nanoparticles of Fe3O4 (MNPs) with β-cyclodextrin (β-CD) on the aggregation/fibrillation of bovine serum albumin (BSA) under diabetic condition known as amyloidogenesis. 26100685 3 46 from conjugation 468:478 arg1 MNP 483:485 MNP 483:485 MNP 483:485 To confirm the β-CD conjugation on MNP, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM) methods were applied. 24719303 3 0 dep spectral 608:615 arg1 overlap 617:623 overlap 617:623 overlap 617:623 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 5 1 theme elusive 1022:1028 arg1 class 1030:1034 this structurally elusive class 1004:1034 this structurally elusive class of biomolecules 1004:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 1 2 theme ROA 118:120 arg1 spectroscopy 123:134 Raman optical activity (ROA) spectroscopy 94:134 Raman optical activity (ROA) spectroscopy 94:134 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 0 3 theme Raman 50:54 arg1 activity 74:81 Raman and Raman optical activity 50:81 Raman and Raman optical activity 50:81 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 5 4 theme biomolecules 1039:1050 arg1 class 1030:1034 this structurally elusive class 1004:1034 this structurally elusive class of biomolecules 1004:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 4 5 theme non-glycosylated 761:776 arg1 isoform 778:784 the non-glycosylated isoform 757:784 the non-glycosylated isoform 757:784 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 4 5 theme non-glycosylated 761:776 arg1 available 789:797 available 789:797 available 789:797 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 2 6 theme moiety 280:285 arg1 data 254:257 Spectral data 245:257 Spectral data of the intact glycan moiety of RNase B 245:296 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 0 7 theme optical 66:72 arg1 activity 74:81 Raman and Raman optical activity 50:81 Raman and Raman optical activity 50:81 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 4 8 theme spectral 693:700 arg1 data 702:705 detailed spectral data 684:705 detailed spectral data 684:705 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 4 9 theme proteins 733:740 arg1 moieties 721:728 the glycan moieties 710:728 the glycan moieties of proteins 710:740 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 3 10 theme remaining 446:454 arg1 spectra 467:473 The remaining difference spectra 442:473 The remaining difference spectra 442:473 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 5 11 theme disaccharide 903:914 arg1 data 916:919 the disaccharide data 899:919 the disaccharide data 899:919 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 1 12 theme protein 211:217 arg1 B 232:232 the protein ribonuclease B 207:232 the protein ribonuclease B (RNase B) 207:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 12 theme protein 211:217 arg1 B 241:241 RNase B 235:241 RNase B 235:241 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 0 13 theme Raman 60:64 arg1 activity 74:81 Raman and Raman optical activity 50:81 Raman and Raman optical activity 50:81 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 3 14 theme difference 456:465 arg1 spectra 467:473 The remaining difference spectra 442:473 The remaining difference spectra 442:473 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 2 15 gly glycoprotein 428:439 arg1 glycoprotein 428:439 the glycoprotein 424:439 the glycoprotein 424:439 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 1 16 theme ribonuclease 219:230 arg1 B 232:232 the protein ribonuclease B 207:232 the protein ribonuclease B (RNase B) 207:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 16 theme ribonuclease 219:230 arg1 B 241:241 RNase B 235:241 RNase B 235:241 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 4 17 theme detailed 684:691 arg1 data 702:705 detailed spectral data 684:705 detailed spectral data 684:705 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 4 18 gly non-glycosylated 761:776 arg1 isoform 778:784 the non-glycosylated isoform 757:784 the non-glycosylated isoform 757:784 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 4 18 gly non-glycosylated 761:776 arg1 available 789:797 available 789:797 available 789:797 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 2 19 theme non-glycosylated 368:383 arg1 form 385:388 the non-glycosylated form 364:388 the non-glycosylated form of the RNase 364:401 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 19 theme non-glycosylated 368:383 arg1 A 361:361 RNase A 355:361 RNase A 355:361 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 5 20 theme glycan 846:851 arg1 spectrum 853:860 the full glycan spectrum 837:860 the full glycan spectrum 837:860 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 2 21 theme RNase 397:401 arg1 form 385:388 the non-glycosylated form 364:388 the non-glycosylated form of the RNase 364:401 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 21 theme RNase 397:401 arg1 A 361:361 RNase A 355:361 RNase A 355:361 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 4 22 theme ROA 648:650 arg1 spectroscopy 652:663 ROA spectroscopy 648:663 ROA spectroscopy 648:663 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 1 23 theme B 232:232 arg1 moiety 197:202 the glycan moiety 186:202 the glycan moiety of the protein ribonuclease B (RNase B) 186:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 2 24 theme Spectral 245:252 arg1 data 254:257 Spectral data 245:257 Spectral data of the intact glycan moiety of RNase B 245:296 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 5 25 theme solution-phase 966:979 arg1 analysis 992:999 the solution-phase structural analysis 962:999 the solution-phase structural analysis of this structurally elusive class of biomolecules 962:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 2 26 theme A 361:361 arg1 data 347:350 high-quality spectral data 325:350 high-quality spectral data of RNase A, the non-glycosylated form of the RNase, 325:402 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 5 27 theme full 841:844 arg1 spectrum 853:860 the full glycan spectrum 837:860 the full glycan spectrum 837:860 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 5 28 theme great 927:931 arg1 tool 953:956 a tool 951:956 a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules 951:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 5 28 theme great 927:931 arg1 promise 933:939 great promise 927:939 great promise to ROA 927:946 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 5 29 theme regenerated 870:880 arg1 spectra 882:888 the regenerated spectra 866:888 the regenerated spectra 866:888 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 2 30 theme RNase 355:359 arg1 form 385:388 the non-glycosylated form 364:388 the non-glycosylated form of the RNase 364:401 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 30 theme RNase 355:359 arg1 A 361:361 RNase A 355:361 RNase A 355:361 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 1 31 theme RNase 235:239 arg1 B 232:232 the protein ribonuclease B 207:232 the protein ribonuclease B (RNase B) 207:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 31 theme RNase 235:239 arg1 B 241:241 RNase B 235:241 RNase B 235:241 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 3 32 theme disaccharides 552:564 arg1 data 528:531 Raman and ROA data 514:531 data 528:531 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 5 33 theme good 813:816 arg1 comparison 818:827 good comparison 813:827 good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data 813:919 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 2 34 theme B 296:296 arg1 data 254:257 Spectral data 245:257 Spectral data of the intact glycan moiety of RNase B 245:296 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 0 35 theme RNase 30:34 arg1 B 36:36 RNase B 30:36 RNase B 30:36 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 2 36 theme spectral 338:345 arg1 data 347:350 high-quality spectral data 325:350 high-quality spectral data of RNase A, the non-glycosylated form of the RNase, 325:402 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 4 37 theme moieties 721:728 arg1 data 702:705 detailed spectral data 684:705 detailed spectral data 684:705 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 1 38 theme solution-phase 158:171 arg1 structure 173:181 the solution-phase structure 154:181 the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B) 154:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 5 39 theme structural 981:990 arg1 analysis 992:999 the solution-phase structural analysis 962:999 the solution-phase structural analysis of this structurally elusive class of biomolecules 962:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 3 40 theme Raman 514:518 arg1 data 528:531 Raman and ROA data 514:531 data 528:531 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 2 41 theme high-quality 325:336 arg1 data 347:350 high-quality spectral data 325:350 high-quality spectral data of RNase A, the non-glycosylated form of the RNase, 325:402 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 42 theme glycoprotein 428:439 arg1 spectra 413:419 the spectra 409:419 the spectra of the glycoprotein 409:439 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 3 43 theme constituent 540:550 arg1 disaccharides 552:564 the constituent disaccharides 536:564 the constituent disaccharides of the RNase glycan 536:584 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 2 44 gly non-glycosylated 368:383 arg1 form 385:388 the non-glycosylated form 364:388 the non-glycosylated form of the RNase 364:401 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 44 gly non-glycosylated 368:383 arg1 RNase 397:401 the RNase 393:401 the RNase 393:401 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 44 gly non-glycosylated 368:383 arg1 A 361:361 RNase A 355:361 RNase A 355:361 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 2 45 theme glycan 273:278 arg1 moiety 280:285 the intact glycan moiety 262:285 the intact glycan moiety 262:285 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 0 46 theme B 36:36 arg1 moiety 20:25 the glycan moiety 9:25 the glycan moiety of RNase B 9:36 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 3 47 theme glycan 579:584 arg1 disaccharides 552:564 the constituent disaccharides 536:564 the constituent disaccharides of the RNase glycan 536:584 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 2 48 theme intact 266:271 arg1 moiety 280:285 the intact glycan moiety 262:285 the intact glycan moiety 262:285 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 3 49 theme ROA 524:526 arg1 data 528:531 Raman and ROA data 514:531 data 528:531 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 4 50 theme glycan 714:719 arg1 moieties 721:728 the glycan moieties 710:728 the glycan moieties of proteins 710:740 The results show that ROA spectroscopy is able to extract detailed spectral data of the glycan moieties of proteins, provided that the non-glycosylated isoform is available. 24719303 1 51 theme Raman 94:98 arg1 spectroscopy 123:134 Raman optical activity (ROA) spectroscopy 94:134 Raman optical activity (ROA) spectroscopy 94:134 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 52 used used 140:143 arg2 Raman 84:88 Raman 84:88 Raman 84:88 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 52 used used 140:143 arg2 spectroscopy 123:134 Raman optical activity (ROA) spectroscopy 94:134 Raman optical activity (ROA) spectroscopy 94:134 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 53 theme optical 100:106 arg1 spectroscopy 123:134 Raman optical activity (ROA) spectroscopy 94:134 Raman optical activity (ROA) spectroscopy 94:134 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 3 54 theme RNase 573:577 arg1 glycan 579:584 the RNase glycan 569:584 the RNase glycan 569:584 The remaining difference spectra are compared to spectra generated from Raman and ROA data of the constituent disaccharides of the RNase glycan, achieving convincing spectral overlap. 24719303 1 55 theme activity 108:115 arg1 spectroscopy 123:134 Raman optical activity (ROA) spectroscopy 94:134 Raman optical activity (ROA) spectroscopy 94:134 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 1 56 theme glycan 190:195 arg1 moiety 197:202 the glycan moiety 186:202 the glycan moiety of the protein ribonuclease B (RNase B) 186:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 24719303 5 57 theme class 1030:1034 arg1 analysis 992:999 the solution-phase structural analysis 962:999 the solution-phase structural analysis of this structurally elusive class of biomolecules 962:1050 Furthermore, good comparison between the full glycan spectrum and the regenerated spectra based on the disaccharide data lends great promise to ROA as a tool for the solution-phase structural analysis of this structurally elusive class of biomolecules. 24719303 0 58 theme glycan 13:18 arg1 moiety 20:25 the glycan moiety 9:25 the glycan moiety of RNase B 9:36 Studying the glycan moiety of RNase B by means of Raman and Raman optical activity. 24719303 2 59 theme RNase 290:294 arg1 B 296:296 RNase B 290:296 RNase B 290:296 Spectral data of the intact glycan moiety of RNase B is obtained by subtracting high-quality spectral data of RNase A, the non-glycosylated form of the RNase, from the spectra of the glycoprotein. 24719303 1 60 theme moiety 197:202 arg1 structure 173:181 the solution-phase structure 154:181 the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B) 154:242 Raman and Raman optical activity (ROA) spectroscopy are used to study the solution-phase structure of the glycan moiety of the protein ribonuclease B (RNase B). 28581490 5 0 dep CCA 855:857 arg1 n=21 860:863 n=21 860:863 n=21 860:863 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 6 1 theme tissue 1150:1155 arg1 areas 1157:1161 MY.1E12-positive tissue areas 1133:1161 MY.1E12-positive tissue areas (2.5 mm2) 1133:1171 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 6 1 theme tissue 1150:1155 arg1 2.5 mm2 1164:1170 2.5 mm2 1164:1170 2.5 mm2 1164:1170 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 10 2 theme M. 1616:1617 arg1 reactivity 1649:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 9 3 theme amurensis 1488:1496 arg1 leukoagglutinin 1498:1512 Maackia amurensis leukoagglutinin and pokeweed lectin 1480:1532 leukoagglutinin 1498:1512 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 5 4 with patients 821:828 arg1 adenocarcinoma 888:901 pancreatic ductal adenocarcinoma 870:901 pancreatic ductal adenocarcinoma (PDAC; n=50) 870:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 4 with patients 821:828 arg1 CCA 855:857 CCA 855:857 CCA 855:857 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 4 with patients 821:828 arg1 cholangiocarcinoma 835:852 cholangiocarcinoma 835:852 cholangiocarcinoma (CCA; n=21) 835:864 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 4 with patients 821:828 arg1 PDAC 904:907 PDAC 904:907 PDAC; n=50 904:913 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 5 theme thick 811:815 arg1 sections 796:803 Tissue sections 789:803 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) 789:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 6 6 theme microdissection 1221:1235 arg1 procedure 1237:1245 a laser capture microdissection procedure 1205:1245 a laser capture microdissection procedure 1205:1245 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 4 7 theme glycoform 676:684 arg1 analysis 686:693 differential glycoform analysis 663:693 differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 663:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 10 8 theme MAH 1644:1646 arg1 reactivity 1649:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 11 9 theme double 1754:1759 arg1 immunostaining 1773:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 These results were confirmed with MAH-MY.1E12 double fluorescent immunostaining. 28581490 13 10 theme novel 2048:2052 arg1 glycodynamics 2054:2066 novel glycodynamics 2048:2066 novel glycodynamics in biology 2048:2077 Our approach provides the information to investigate novel glycodynamics in biology, for example, glycoalteration, as well as diseases related to not only MUC1 but also other membrane proteins. 28581490 12 11 theme high 1837:1840 arg1 sensitivity 1842:1852 high sensitivity 1837:1852 high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1837:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 6 12 theme laser 1207:1211 arg1 microdissection 1221:1235 a laser capture microdissection 1205:1235 a laser capture microdissection procedure 1205:1245 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 9 13 theme MY.1E12-reactive 1548:1563 arg1 CCA 1573:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 4 14 theme possible 643:650 arg1 method 652:657 a possible method 641:657 a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 641:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 5 15 theme NeuAcα2-3galactosyl 1084:1102 arg1 structure 1073:1081 a sialyl-core 1 structure 1057:1081 a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 1057:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 15 theme NeuAcα2-3galactosyl 1084:1102 arg1 β1-3-N-acetylgalactosamine 1104:1129 NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine 1084:1129 NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine 1084:1129 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 9 16 theme of 1570:1571 arg1 CCA 1573:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 9 17 dep found 1474:1478 arg1 distinguished 1534:1546 distinguished 1534:1546 distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC 1534:1593 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 3 18 theme modifications 429:441 arg1 one 408:410 one 408:410 one 408:410 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 3 18 theme modifications 429:441 arg1 modifications 429:441 the candidate modifications 415:441 the candidate modifications in a protein 415:454 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 3 19 theme sensitivity 608:618 arg1 lack 589:592 the lack 585:592 the lack of analytical sensitivity 585:618 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 5 20 theme pancreatic 870:879 arg1 adenocarcinoma 888:901 pancreatic ductal adenocarcinoma 870:901 pancreatic ductal adenocarcinoma (PDAC; n=50) 870:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 20 theme pancreatic 870:879 arg1 PDAC 904:907 PDAC 904:907 PDAC; n=50 904:913 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 10 21 theme malignancy 1696:1705 arg1 degree 1686:1691 the degree 1682:1691 the degree of malignancy 1682:1705 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 5 22 theme sialyl-core 1059:1069 arg1 structure 1073:1081 a sialyl-core 1 structure 1057:1081 a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 1057:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 22 theme sialyl-core 1059:1069 arg1 β1-3-N-acetylgalactosamine 1104:1129 NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine 1084:1129 NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine 1084:1129 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 4 23 theme antibody-overlay 753:768 arg1 microarray 777:786 an antibody-overlay lectin microarray 750:786 an antibody-overlay lectin microarray 750:786 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 4 24 gly glycoprotein 729:740 arg1 glycoprotein 729:740 an endogenous tumor-associated glycoprotein MUC1 698:745 an endogenous tumor-associated glycoprotein MUC1 698:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 2 25 theme diagnostic 249:258 arg1 specificity 260:270 diagnostic specificity 249:270 diagnostic specificity in molecular pathology 249:293 In general, diagnostic specificity in molecular pathology is increased by the acquisition of a probe to distinguish the modification of isomers. 28581490 5 26 theme anti-MUC1 937:945 arg1 antibody 947:954 an anti-MUC1 antibody MY.1E12 934:962 an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 934:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 26 theme anti-MUC1 937:945 arg1 antibody 1001:1008 a monoclonal antibody 988:1008 a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 988:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 7 27 with immunoprecipitation 1282:1300 arg1 MY.1E12 1307:1313 MY.1E12 1307:1313 MY.1E12 1307:1313 The membrane MUC1 was enriched by immunoprecipitation with MY.1E12 and subjected to lectin microarray analysis. 28581490 8 28 theme MY.1E12 1392:1398 arg1 reactivities 1376:1387 the reactivities 1372:1387 the reactivities of MY.1E12 between CCA and PDAC 1372:1419 Even though the reactivities of MY.1E12 between CCA and PDAC were similar, the lectin-binding patterns varied. 28581490 8 28 theme MY.1E12 1392:1398 arg1 similar 1426:1432 similar 1426:1432 similar 1426:1432 Even though the reactivities of MY.1E12 between CCA and PDAC were similar, the lectin-binding patterns varied. 28581490 12 29 theme glycan 1795:1800 arg1 changes 1802:1808 These glycan changes 1789:1808 These glycan changes on MUC1 1789:1816 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 4 30 theme glycoprotein 729:740 arg1 MUC1 742:745 an endogenous tumor-associated glycoprotein MUC1 698:745 an endogenous tumor-associated glycoprotein MUC1 698:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 0 31 theme MUC1 85:88 arg1 analysis 55:62 glycoform analysis 45:62 glycoform analysis of membrane-tethered MUC1 45:88 Assessment of tumor characteristics based on glycoform analysis of membrane-tethered MUC1. 28581490 13 32 theme membrane 2170:2177 arg1 proteins 2179:2186 other membrane proteins 2164:2186 not only MUC1 but also other membrane proteins 2141:2186 Our approach provides the information to investigate novel glycodynamics in biology, for example, glycoalteration, as well as diseases related to not only MUC1 but also other membrane proteins. 28581490 13 33 from glycodynamics 2054:2066 arg1 biology 2071:2077 biology 2071:2077 biology 2071:2077 Our approach provides the information to investigate novel glycodynamics in biology, for example, glycoalteration, as well as diseases related to not only MUC1 but also other membrane proteins. 28581490 5 34 theme Tissue 789:794 arg1 sections 796:803 Tissue sections 789:803 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) 789:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 35 theme monoclonal 990:999 arg1 antibody 1001:1008 a monoclonal antibody 988:1008 a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 988:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 35 theme monoclonal 990:999 arg1 antibody 947:954 an anti-MUC1 antibody MY.1E12 934:962 an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine) 934:1130 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 12 36 from effect 1875:1880 arg1 antigen 1932:1938 a tandem repeat peptide antigen 1908:1938 a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1908:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 3 37 theme candidate 419:427 arg1 modifications 429:441 the candidate modifications 415:441 the candidate modifications in a protein 415:454 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 1 38 theme tissue 100:105 arg1 specimens 107:115 Clinical tissue specimens 91:115 Clinical tissue specimens 91:115 Clinical tissue specimens are useful for pathological diagnosis, which is, in some cases, supported by visualization of biomolecule localization. 28581490 3 39 theme comparative 457:467 arg1 analysis 476:483 comparative glycan analysis 457:483 comparative glycan analysis of disease-associated proteins derived from a single tissue section 457:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 0 40 theme tumor 14:18 arg1 characteristics 20:34 tumor characteristics 14:34 tumor characteristics 14:34 Assessment of tumor characteristics based on glycoform analysis of membrane-tethered MUC1. 28581490 3 41 theme tissue 538:543 arg1 section 545:551 a single tissue section 529:551 a single tissue section 529:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 2 42 from specificity 260:270 arg1 pathology 285:293 molecular pathology 275:293 molecular pathology 275:293 In general, diagnostic specificity in molecular pathology is increased by the acquisition of a probe to distinguish the modification of isomers. 28581490 4 43 theme endogenous 701:710 arg1 MUC1 742:745 an endogenous tumor-associated glycoprotein MUC1 698:745 an endogenous tumor-associated glycoprotein MUC1 698:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 5 44 theme ductal 881:886 arg1 adenocarcinoma 888:901 pancreatic ductal adenocarcinoma 870:901 pancreatic ductal adenocarcinoma (PDAC; n=50) 870:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 44 theme ductal 881:886 arg1 PDAC 904:907 PDAC 904:907 PDAC; n=50 904:913 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 12 45 theme repeat 1917:1922 arg1 antigen 1932:1938 a tandem repeat peptide antigen 1908:1938 a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1908:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 3 46 theme disease-associated 488:505 arg1 proteins 507:514 disease-associated proteins 488:514 disease-associated proteins derived from a single tissue section 488:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 0 47 theme glycoform 45:53 arg1 analysis 55:62 glycoform analysis 45:62 glycoform analysis of membrane-tethered MUC1 45:88 Assessment of tumor characteristics based on glycoform analysis of membrane-tethered MUC1. 28581490 1 48 theme biomolecule 211:221 arg1 localization 223:234 biomolecule localization 211:234 biomolecule localization 211:234 Clinical tissue specimens are useful for pathological diagnosis, which is, in some cases, supported by visualization of biomolecule localization. 28581490 12 49 theme lectins 1897:1903 arg1 effect 1875:1880 the cluster effect 1863:1880 the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1863:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 7 50 theme microarray 1339:1348 arg1 analysis 1350:1357 lectin microarray analysis 1332:1357 lectin microarray analysis 1332:1357 The membrane MUC1 was enriched by immunoprecipitation with MY.1E12 and subjected to lectin microarray analysis. 28581490 3 51 from modifications 429:441 arg1 protein 448:454 a protein 446:454 a protein 446:454 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 5 52 theme MUC1 1025:1028 arg1 isoform 1044:1050 an MUC1 glycosylation isoform 1022:1050 an MUC1 glycosylation isoform 1022:1050 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 10 53 theme amurensis 1619:1627 arg1 reactivity 1649:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 11 54 theme MAH-MY.1E12 1742:1752 arg1 immunostaining 1773:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 These results were confirmed with MAH-MY.1E12 double fluorescent immunostaining. 28581490 9 55 theme Maackia 1480:1486 arg1 leukoagglutinin 1498:1512 Maackia amurensis leukoagglutinin and pokeweed lectin 1480:1532 leukoagglutinin 1498:1512 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 12 56 theme cluster 1867:1873 arg1 effect 1875:1880 the cluster effect 1863:1880 the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1863:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 6 57 theme MY.1E12-positive 1133:1148 arg1 areas 1157:1161 MY.1E12-positive tissue areas 1133:1161 MY.1E12-positive tissue areas (2.5 mm2) 1133:1171 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 6 57 theme MY.1E12-positive 1133:1148 arg1 2.5 mm2 1164:1170 2.5 mm2 1164:1170 2.5 mm2 1164:1170 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 2 58 theme molecular 275:283 arg1 pathology 285:293 molecular pathology 275:293 molecular pathology 275:293 In general, diagnostic specificity in molecular pathology is increased by the acquisition of a probe to distinguish the modification of isomers. 28581490 9 59 theme pokeweed 1518:1525 arg1 lectin 1527:1532 Maackia amurensis leukoagglutinin and pokeweed lectin 1480:1532 lectin 1527:1532 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 6 60 theme capture 1213:1219 arg1 microdissection 1221:1235 a laser capture microdissection 1205:1235 a laser capture microdissection procedure 1205:1245 MY.1E12-positive tissue areas (2.5 mm2) were selectively dissected with a laser capture microdissection procedure. 28581490 4 61 theme differential 663:674 arg1 analysis 686:693 differential glycoform analysis 663:693 differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 663:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 11 62 theme fluorescent 1761:1771 arg1 immunostaining 1773:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 MAH-MY.1E12 double fluorescent immunostaining 1742:1786 These results were confirmed with MAH-MY.1E12 double fluorescent immunostaining. 28581490 9 63 theme MUC1 1565:1568 arg1 CCA 1573:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 MY.1E12-reactive MUC1 of CCA 1548:1575 We found Maackia amurensis leukoagglutinin and pokeweed lectin distinguished MY.1E12-reactive MUC1 of CCA from that of PDAC. 28581490 2 64 theme probe 332:336 arg1 acquisition 315:325 the acquisition 311:325 the acquisition of a probe to distinguish the modification of isomers 311:379 In general, diagnostic specificity in molecular pathology is increased by the acquisition of a probe to distinguish the modification of isomers. 28581490 3 65 theme analytical 597:606 arg1 sensitivity 608:618 analytical sensitivity 597:618 analytical sensitivity 597:618 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 4 66 theme MUC1 742:745 arg1 analysis 686:693 differential glycoform analysis 663:693 differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 663:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 1 67 theme pathological 132:143 arg1 diagnosis 145:153 pathological diagnosis 132:153 pathological diagnosis 132:153 Clinical tissue specimens are useful for pathological diagnosis, which is, in some cases, supported by visualization of biomolecule localization. 28581490 3 68 attach derived 516:522 arg2 proteins 507:514 disease-associated proteins 488:514 disease-associated proteins derived from a single tissue section 488:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 3 68 attach derived 516:522 arg1 section 545:551 a single tissue section 529:551 a single tissue section 529:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 12 69 from changes 1802:1808 arg1 MUC1 1813:1816 MUC1 1813:1816 MUC1 1813:1816 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 0 70 theme membrane-tethered 67:83 arg1 MUC1 85:88 membrane-tethered MUC1 67:88 membrane-tethered MUC1 67:88 Assessment of tumor characteristics based on glycoform analysis of membrane-tethered MUC1. 28581490 13 71 theme other 2164:2168 arg1 proteins 2179:2186 other membrane proteins 2164:2186 not only MUC1 but also other membrane proteins 2141:2186 Our approach provides the information to investigate novel glycodynamics in biology, for example, glycoalteration, as well as diseases related to not only MUC1 but also other membrane proteins. 28581490 4 72 theme tumor-associated 712:727 arg1 MUC1 742:745 an endogenous tumor-associated glycoprotein MUC1 698:745 an endogenous tumor-associated glycoprotein MUC1 698:745 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 10 73 with MUC1 1606:1609 arg1 reactivity 1649:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 1 74 theme Clinical 91:98 arg1 specimens 107:115 Clinical tissue specimens 91:115 Clinical tissue specimens 91:115 Clinical tissue specimens are useful for pathological diagnosis, which is, in some cases, supported by visualization of biomolecule localization. 28581490 4 75 theme lectin 770:775 arg1 microarray 777:786 an antibody-overlay lectin microarray 750:786 an antibody-overlay lectin microarray 750:786 Here we demonstrate a possible method for differential glycoform analysis of an endogenous tumor-associated glycoprotein MUC1 by an antibody-overlay lectin microarray. 28581490 2 76 theme isomers 373:379 arg1 modification 357:368 the modification 353:368 the modification of isomers 353:379 In general, diagnostic specificity in molecular pathology is increased by the acquisition of a probe to distinguish the modification of isomers. 28581490 0 77 theme characteristics 20:34 arg1 Assessment 0:9 Assessment 0:9 Assessment of tumor characteristics 0:34 Assessment of tumor characteristics based on glycoform analysis of membrane-tethered MUC1. 28581490 8 78 theme lectin-binding 1439:1452 arg1 patterns 1454:1461 the lectin-binding patterns 1435:1461 the lectin-binding patterns 1435:1461 Even though the reactivities of MY.1E12 between CCA and PDAC were similar, the lectin-binding patterns varied. 28581490 3 79 theme glycan 469:474 arg1 analysis 476:483 comparative glycan analysis 457:483 comparative glycan analysis of disease-associated proteins derived from a single tissue section 457:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 5 80 theme glycosylation 1030:1042 arg1 isoform 1044:1050 an MUC1 glycosylation isoform 1022:1050 an MUC1 glycosylation isoform 1022:1050 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 5 81 dep PDAC 904:907 arg1 n=50 910:913 n=50 910:913 PDAC; n=50 904:913 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 7 82 theme membrane 1252:1259 arg1 MUC1 1261:1264 The membrane MUC1 1248:1264 The membrane MUC1 1248:1264 The membrane MUC1 was enriched by immunoprecipitation with MY.1E12 and subjected to lectin microarray analysis. 28581490 5 83 theme patients 821:828 arg1 sections 796:803 Tissue sections 789:803 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) 789:914 Tissue sections (5 μm thick) of patients with cholangiocarcinoma (CCA; n=21) and pancreatic ductal adenocarcinoma (PDAC; n=50) were stained with an anti-MUC1 antibody MY.1E12 that was established as a monoclonal antibody recognizing an MUC1 glycosylation isoform with a sialyl-core 1 structure (NeuAcα2-3galactosyl β1-3-N-acetylgalactosamine). 28581490 12 84 theme tandem 1910:1915 arg1 antigen 1932:1938 a tandem repeat peptide antigen 1908:1938 a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1908:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 13 85 theme related 2130:2136 arg1 diseases 2121:2128 diseases 2121:2128 diseases related to not only MUC1 but also other membrane proteins 2121:2186 Our approach provides the information to investigate novel glycodynamics in biology, for example, glycoalteration, as well as diseases related to not only MUC1 but also other membrane proteins. 28581490 3 86 theme proteins 507:514 arg1 analysis 476:483 comparative glycan analysis 457:483 comparative glycan analysis of disease-associated proteins derived from a single tissue section 457:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 28581490 12 87 theme dense 1960:1964 arg1 glycosylation 1966:1978 highly dense glycosylation 1953:1978 highly dense glycosylation such as mucin 1953:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 12 88 theme peptide 1924:1930 arg1 antigen 1932:1938 a tandem repeat peptide antigen 1908:1938 a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin 1908:1992 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 7 89 theme lectin 1332:1337 arg1 analysis 1350:1357 lectin microarray analysis 1332:1357 lectin microarray analysis 1332:1357 The membrane MUC1 was enriched by immunoprecipitation with MY.1E12 and subjected to lectin microarray analysis. 28581490 1 90 theme localization 223:234 arg1 visualization 194:206 visualization 194:206 visualization of biomolecule localization 194:234 Clinical tissue specimens are useful for pathological diagnosis, which is, in some cases, supported by visualization of biomolecule localization. 28581490 12 91 theme immobilized 1885:1895 arg1 lectins 1897:1903 immobilized lectins 1885:1903 immobilized lectins 1885:1903 These glycan changes on MUC1 were detected with high sensitivity owing to the cluster effect of immobilized lectins on a tandem repeat peptide antigen covered with highly dense glycosylation such as mucin. 28581490 10 92 theme hemagglutinin 1629:1641 arg1 reactivity 1649:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 M. amurensis hemagglutinin (MAH) reactivity 1616:1658 Moreover, MUC1 with M. amurensis hemagglutinin (MAH) reactivity potentially reflected the degree of malignancy. 28581490 3 93 theme single 531:536 arg1 section 545:551 a single tissue section 529:551 a single tissue section 529:551 Although glycosylation is one of the candidate modifications in a protein, comparative glycan analysis of disease-associated proteins derived from a single tissue section is still challenging because of the lack of analytical sensitivity. 25139566 0 0 theme N-acetylglucosaminyltransferase 83:113 arg1 requirements 67:78 substrate structural requirements 46:78 substrate structural requirements of N-acetylglucosaminyltransferase III 46:117 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 7 1 theme GnT-III 1042:1048 arg1 assay 1061:1065 A competitive GnT-III inhibition assay 1028:1065 A competitive GnT-III inhibition assay using the synthetic substrates 1028:1096 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 4 2 theme substrate-binding 623:639 arg1 mode 641:644 even the substrate-binding mode 614:644 even the substrate-binding mode of GnT-III 614:655 Thus far, however, such inhibitors have not been developed and even the substrate-binding mode of GnT-III is not fully understood. 25139566 2 3 theme proteins 406:413 arg1 regulation 378:387 the functional regulation 363:387 the functional regulation of their carrier proteins 363:413 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 5 4 theme structural 703:712 arg1 features 714:721 structural features 703:721 structural features required of the substrate 703:747 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 2 5 theme pathological 332:343 arg1 processes 345:353 physiological and pathological processes 314:353 physiological and pathological processes 314:353 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 5 6 theme N-glycan 785:792 arg1 units 794:798 four N-glycan units 780:798 four N-glycan units 780:798 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 5 6 theme N-glycan 785:792 arg1 parts 815:819 the branching parts 801:819 the branching parts of the bisected and non-bisected N-glycans 801:862 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 9 7 theme rational 1338:1345 arg1 design 1357:1362 rational inhibitor design 1338:1362 rational inhibitor design 1338:1362 The structural requirements of GnT-III elucidated in this study will provide a basis for rational inhibitor design. 25139566 8 8 theme previous 1168:1175 arg1 reports 1177:1183 previous reports 1168:1183 previous reports 1168:1183 In keeping with previous reports, GlcNAc at the α1,3-branch is also involved in the interaction. 25139566 9 9 theme inhibitor 1347:1355 arg1 design 1357:1362 rational inhibitor design 1338:1362 rational inhibitor design 1338:1362 The structural requirements of GnT-III elucidated in this study will provide a basis for rational inhibitor design. 25139566 7 10 theme GlcNAc 1137:1142 arg1 moiety 1144:1149 the Manβ(1-4)GlcNAc moiety 1124:1149 the Manβ(1-4)GlcNAc moiety 1124:1149 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 2 11 theme physiological 314:326 arg1 processes 345:353 physiological and pathological processes 314:353 physiological and pathological processes 314:353 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 1 12 theme bisected 202:209 arg1 N-glycans 211:219 bisected N-glycans 202:219 bisected N-glycans 202:219 N-Acetylglucosaminyltransferase (GnT) III is a glycosyltransferase which produces bisected N-glycans by transferring GlcNAc to the 4-position of core mannose. 25139566 5 13 theme non-bisected 841:852 arg1 N-glycans 854:862 the bisected and non-bisected N-glycans 824:862 N-glycans 854:862 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 4 14 theme GnT-III 649:655 arg1 mode 641:644 even the substrate-binding mode 614:644 even the substrate-binding mode of GnT-III 614:655 Thus far, however, such inhibitors have not been developed and even the substrate-binding mode of GnT-III is not fully understood. 25139566 3 15 theme biological 440:449 arg1 functions 451:459 the biological functions 436:459 the biological functions of bisected glycans 436:479 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 2 16 theme functional 367:376 arg1 regulation 378:387 the functional regulation 363:387 the functional regulation of their carrier proteins 363:413 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 3 17 theme functions 451:459 arg1 understanding 419:431 An understanding 416:431 An understanding of the biological functions of bisected glycans 416:479 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 0 18 theme units 22:26 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 0:118 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 7 19 theme Manβ 1128:1131 arg1 moiety 1144:1149 the Manβ(1-4)GlcNAc moiety 1124:1149 the Manβ(1-4)GlcNAc moiety 1124:1149 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 9 20 theme structural 1253:1262 arg1 requirements 1264:1275 The structural requirements 1249:1275 The structural requirements of GnT-III elucidated in this study 1249:1311 The structural requirements of GnT-III elucidated in this study will provide a basis for rational inhibitor design. 25139566 3 21 theme specific 519:526 arg1 inhibitors 528:537 specific inhibitors 519:537 specific inhibitors of GnT-III 519:548 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 0 22 theme N-glycan 13:20 arg1 units 22:26 N-glycan units 13:26 N-glycan units 13:26 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 6 23 theme core 917:920 arg1 trimannose 922:931 a common core trimannose 908:931 a common core trimannose 908:931 The series of syntheses were achieved from a common core trimannose, giving bisected tetra- and hexasaccharides as well as non-bisected tri- and pentasaccharides. 25139566 3 24 theme inhibitors 528:537 arg1 use 512:514 use 512:514 use of specific inhibitors of GnT-III 512:548 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 3 25 theme glycans 473:479 arg1 functions 451:459 the biological functions 436:459 the biological functions of bisected glycans 436:479 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 6 26 theme common 910:915 arg1 trimannose 922:931 a common core trimannose 908:931 a common core trimannose 908:931 The series of syntheses were achieved from a common core trimannose, giving bisected tetra- and hexasaccharides as well as non-bisected tri- and pentasaccharides. 25139566 5 27 theme bisected 828:835 arg1 N-glycans 854:862 the bisected and non-bisected N-glycans 824:862 N-glycans 854:862 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 9 28 theme GnT-III 1280:1286 arg1 requirements 1264:1275 The structural requirements 1249:1275 The structural requirements of GnT-III elucidated in this study 1249:1311 The structural requirements of GnT-III elucidated in this study will provide a basis for rational inhibitor design. 25139566 7 29 theme synthetic 1077:1085 arg1 substrates 1087:1096 the synthetic substrates 1073:1096 the synthetic substrates 1073:1096 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 0 30 theme substrate 46:54 arg1 requirements 67:78 substrate structural requirements 46:78 substrate structural requirements of N-acetylglucosaminyltransferase III 46:117 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 4 31 theme such 570:573 arg1 inhibitors 575:584 such inhibitors 570:584 such inhibitors 570:584 Thus far, however, such inhibitors have not been developed and even the substrate-binding mode of GnT-III is not fully understood. 25139566 7 32 theme vital 1109:1113 arg1 role 1115:1118 a vital role 1107:1118 a vital role for the Manβ(1-4)GlcNAc moiety 1107:1149 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 5 33 theme branching 805:813 arg1 units 794:798 four N-glycan units 780:798 four N-glycan units 780:798 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 5 33 theme branching 805:813 arg1 parts 815:819 the branching parts 801:819 the branching parts of the bisected and non-bisected N-glycans 801:862 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 2 34 theme carrier 398:404 arg1 proteins 406:413 their carrier proteins 392:413 their carrier proteins 392:413 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 3 35 theme bisected 464:471 arg1 glycans 473:479 bisected glycans 464:479 bisected glycans 464:479 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 8 36 from α1,3-branch 1200:1210 arg1 GlcNAc 1186:1191 GlcNAc 1186:1191 GlcNAc at the α1,3-branch 1186:1210 In keeping with previous reports, GlcNAc at the α1,3-branch is also involved in the interaction. 25139566 6 37 theme syntheses 879:887 arg1 series 869:874 The series 865:874 The series of syntheses 865:887 The series of syntheses were achieved from a common core trimannose, giving bisected tetra- and hexasaccharides as well as non-bisected tri- and pentasaccharides. 25139566 7 38 theme inhibition 1050:1059 arg1 assay 1061:1065 A competitive GnT-III inhibition assay 1028:1065 A competitive GnT-III inhibition assay using the synthetic substrates 1028:1096 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 3 39 theme GnT-III 542:548 arg1 inhibitors 528:537 specific inhibitors 519:537 specific inhibitors of GnT-III 519:548 An understanding of the biological functions of bisected glycans will be greatly accelerated by use of specific inhibitors of GnT-III. 25139566 1 40 theme core 265:268 arg1 mannose 270:276 core mannose 265:276 core mannose 265:276 N-Acetylglucosaminyltransferase (GnT) III is a glycosyltransferase which produces bisected N-glycans by transferring GlcNAc to the 4-position of core mannose. 25139566 0 41 theme requirements 67:78 arg1 assessment 32:41 assessment 32:41 assessment of substrate structural requirements of N-acetylglucosaminyltransferase III 32:117 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 1 42 theme mannose 270:276 arg1 4-position 251:260 the 4-position 247:260 the 4-position of core mannose 247:276 N-Acetylglucosaminyltransferase (GnT) III is a glycosyltransferase which produces bisected N-glycans by transferring GlcNAc to the 4-position of core mannose. 25139566 0 43 theme structural 56:65 arg1 requirements 67:78 substrate structural requirements 46:78 substrate structural requirements of N-acetylglucosaminyltransferase III 46:117 Synthesis of N-glycan units for assessment of substrate structural requirements of N-acetylglucosaminyltransferase III. 25139566 2 44 theme Bisected 279:286 arg1 N-glycans 288:296 Bisected N-glycans 279:296 Bisected N-glycans 279:296 Bisected N-glycans are involved in physiological and pathological processes through the functional regulation of their carrier proteins. 25139566 7 45 theme competitive 1030:1040 arg1 assay 1061:1065 A competitive GnT-III inhibition assay 1028:1065 A competitive GnT-III inhibition assay using the synthetic substrates 1028:1096 A competitive GnT-III inhibition assay using the synthetic substrates revealed a vital role for the Manβ(1-4)GlcNAc moiety. 25139566 5 46 theme N-glycans 854:862 arg1 units 794:798 four N-glycan units 780:798 four N-glycan units 780:798 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 5 46 theme N-glycans 854:862 arg1 parts 815:819 the branching parts 801:819 the branching parts of the bisected and non-bisected N-glycans 801:862 To gain insight into structural features required of the substrate, we systematically synthesized four N-glycan units, the branching parts of the bisected and non-bisected N-glycans. 25139566 6 47 theme non-bisected 988:999 arg1 tri- 1001:1004 non-bisected tri- 988:1004 non-bisected tri- 988:1004 The series of syntheses were achieved from a common core trimannose, giving bisected tetra- and hexasaccharides as well as non-bisected tri- and pentasaccharides. 28056108 9 0 theme model 1568:1572 arg1 glycoprotein 1589:1600 a model N-glycosylated glycoprotein 1566:1600 a model N-glycosylated glycoprotein 1566:1600 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 8 1 theme binding 1354:1360 arg1 protein 1362:1368 the solute binding protein 1343:1368 the solute binding protein of an ABC transporter (ABCNG) 1343:1398 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 1 theme binding 1354:1360 arg1 NgtS 1337:1340 NgtS 1337:1340 NgtS 1337:1340 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 11 2 theme full 1903:1906 arg1 virulence 1908:1916 full virulence 1903:1916 full virulence in S. pneumoniae 1903:1933 Therefore, N-glycan processing, but not transport of the released glycan, is required for full virulence in S. pneumoniae. 28056108 12 3 theme processing 2001:2010 arg1 model 1983:1987 a model 1981:1987 a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG 1981:2162 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 3 4 theme GlcNAc 749:754 arg1 residues 756:763 the two GlcNAc residues 741:763 the two GlcNAc residues 741:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 3 5 with N-glycans 530:538 arg1 enzymes 545:551 enzymes 545:551 enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 545:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 3 6 theme prior 624:628 arg1 core 619:622 the Man3GlcNAc2 core 603:622 the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 603:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 3 7 theme Man3GlcNAc2 607:617 arg1 core 619:622 the Man3GlcNAc2 core 603:622 the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 603:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 5 8 theme biochemical 882:892 arg1 analyses 909:916 biochemical and structural analyses 882:916 biochemical and structural analyses 882:916 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 1 9 theme carbohydrate-rich 132:148 arg1 coating 150:156 The carbohydrate-rich coating 128:156 The carbohydrate-rich coating of human tissues and cells 128:183 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 2 10 contain possess 413:419 arg1 bacteria 404:411 some host-adapted bacteria 386:411 some host-adapted bacteria 386:411 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 10 contain possess 413:419 arg2 machinery 425:433 the machinery to process N-linked glycans 421:461 the machinery to process N-linked glycans 421:461 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 5 11 theme structural 898:907 arg1 analyses 909:916 biochemical and structural analyses 882:916 biochemical and structural analyses 882:916 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 12 theme α- 970:971 arg1 SpGH92 991:996 SpGH92 991:996 SpGH92 991:996 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 12 theme α- 970:971 arg1 -mannosidase 977:988 an α-(1,2)-mannosidase 967:988 an α-(1,2)-mannosidase (SpGH92) 967:997 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 12 theme α- 970:971 arg1 1,2 973:975 1,2 973:975 1,2 973:975 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 7 13 link -linked 1259:1265 arg1 residues 1275:1282 α-(1,2)-linked mannose residues 1252:1282 α-(1,2)-linked mannose residues 1252:1282 Through this activity SpGH92 is able to produce a substrate for EndoD, which is not active on high-mannose glycans with α-(1,2)-linked mannose residues. 28056108 10 14 theme pneumoniae 1745:1754 arg1 growth 1732:1737 growth 1732:1737 growth of S. pneumoniae 1732:1754 We found that both EndoD and ABCNG contribute to growth of S. pneumoniae, but that only SpGH92 and EndoD contribute to virulence. 28056108 6 15 theme terminal 1040:1047 arg1 residues 1072:1079 the terminal α-(1,2)-linked mannose residues 1036:1079 the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans 1036:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 15 theme terminal 1040:1047 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 9 16 theme mouse 1670:1674 arg1 model 1676:1680 a mouse model 1668:1680 a mouse model 1668:1680 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 0 17 from Contribution 101:112 arg1 pneumoniae 82:91 Streptococcus pneumoniae 68:91 Streptococcus pneumoniae 68:91 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 8 18 theme X-ray 1305:1309 arg1 crystallography 1311:1325 X-ray crystallography 1305:1325 X-ray crystallography 1305:1325 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 7 19 theme high-mannose 1226:1237 arg1 glycans 1239:1245 high-mannose glycans 1226:1245 high-mannose glycans with α-(1,2)-linked mannose residues 1226:1282 Through this activity SpGH92 is able to produce a substrate for EndoD, which is not active on high-mannose glycans with α-(1,2)-linked mannose residues. 28056108 11 20 theme released 1870:1877 arg1 glycan 1879:1884 the released glycan 1866:1884 the released glycan 1866:1884 Therefore, N-glycan processing, but not transport of the released glycan, is required for full virulence in S. pneumoniae. 28056108 1 21 theme tissues 167:173 arg1 coating 150:156 The carbohydrate-rich coating 128:156 The carbohydrate-rich coating of human tissues and cells 128:183 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 8 22 theme EndoD 1442:1446 arg1 activity 1448:1455 EndoD activity 1442:1455 EndoD activity 1442:1455 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 1 23 theme cells 179:183 arg1 coating 150:156 The carbohydrate-rich coating 128:156 The carbohydrate-rich coating of human tissues and cells 128:183 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 9 24 from SpGH92 1645:1650 arg1 model 1676:1680 a mouse model 1668:1680 a mouse model 1668:1680 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 0 25 theme Contribution 101:112 arg1 Characterization 10:25 Molecular Characterization 0:25 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 0:126 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 0 26 from Transport 55:63 arg1 pneumoniae 82:91 Streptococcus pneumoniae 68:91 Streptococcus pneumoniae 68:91 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 0 27 from Degradation 39:49 arg1 pneumoniae 82:91 Streptococcus pneumoniae 68:91 Streptococcus pneumoniae 68:91 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 6 28 contain has 1012:1014 arg2 ability 1020:1026 the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2 1016:1129 the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2 1016:1129 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 28 contain has 1012:1014 arg1 enzyme 1005:1010 This enzyme 1000:1010 This enzyme 1000:1010 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 0 29 theme Molecular 0:8 arg1 Characterization 10:25 Molecular Characterization 0:25 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 0:126 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 6 30 theme mannose 1064:1070 arg1 residues 1072:1079 the terminal α-(1,2)-linked mannose residues 1036:1079 the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans 1036:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 30 theme mannose 1064:1070 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 2 31 theme N-linked 446:453 arg1 glycans 455:461 N-linked glycans 446:461 N-linked glycans 446:461 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 3 32 theme complex 578:584 arg1 N-glycan 586:593 a complex N-glycan 576:593 a complex N-glycan 576:593 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 8 33 theme ABC 1376:1378 arg1 transporter 1380:1390 an ABC transporter 1373:1390 an ABC transporter (ABCNG) 1373:1398 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 33 theme ABC 1376:1378 arg1 ABCNG 1393:1397 ABCNG 1393:1397 ABCNG 1393:1397 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 3 34 theme pathogen 474:481 arg1 pneumoniae 497:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 0 35 theme Degradation 39:49 arg1 Characterization 10:25 Molecular Characterization 0:25 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 0:126 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 0 36 theme Streptococcus 68:80 arg1 pneumoniae 82:91 Streptococcus pneumoniae 68:91 Streptococcus pneumoniae 68:91 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 6 37 link -linked 1056:1062 arg1 residues 1072:1079 the terminal α-(1,2)-linked mannose residues 1036:1079 the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans 1036:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 37 link -linked 1056:1062 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 0 38 theme Transport 55:63 arg1 Characterization 10:25 Molecular Characterization 0:25 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 0:126 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 7 39 theme mannose 1267:1273 arg1 residues 1275:1282 α-(1,2)-linked mannose residues 1252:1282 α-(1,2)-linked mannose residues 1252:1282 Through this activity SpGH92 is able to produce a substrate for EndoD, which is not active on high-mannose glycans with α-(1,2)-linked mannose residues. 28056108 11 40 from virulence 1908:1916 arg1 pneumoniae 1924:1933 S. pneumoniae 1921:1933 S. pneumoniae 1921:1933 Therefore, N-glycan processing, but not transport of the released glycan, is required for full virulence in S. pneumoniae. 28056108 3 41 theme glycan 652:657 arg1 release 637:643 the release 633:643 the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 633:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 4 42 theme high-mannose 823:834 arg1 N-glycans 836:844 high-mannose N-glycans 823:844 high-mannose N-glycans 823:844 Here we examine the capacity of S. pneumoniae to process high-mannose N-glycans and transport the products. 28056108 3 43 theme complex 522:528 arg1 N-glycans 530:538 complex N-glycans 522:538 complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 522:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 9 44 theme pneumoniae 1552:1561 arg1 growth 1539:1544 growth 1539:1544 growth of S. pneumoniae 1539:1561 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 11 45 theme N-glycan 1824:1831 arg1 processing 1833:1842 N-glycan processing 1824:1842 N-glycan processing 1824:1842 Therefore, N-glycan processing, but not transport of the released glycan, is required for full virulence in S. pneumoniae. 28056108 9 46 gly glycoprotein 1589:1600 arg1 glycoprotein 1589:1600 a model N-glycosylated glycoprotein 1566:1600 a model N-glycosylated glycoprotein 1566:1600 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 7 47 with glycans 1239:1245 arg1 residues 1275:1282 α-(1,2)-linked mannose residues 1252:1282 α-(1,2)-linked mannose residues 1252:1282 Through this activity SpGH92 is able to produce a substrate for EndoD, which is not active on high-mannose glycans with α-(1,2)-linked mannose residues. 28056108 12 48 theme pathway 2138:2144 arg1 arms 2113:2116 the two arms 2105:2116 the two arms of this degradation pathway 2105:2144 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 12 48 theme pathway 2138:2144 arg1 pathway 2138:2144 this degradation pathway 2121:2144 this degradation pathway 2121:2144 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 9 49 theme N-glycosylated 1574:1587 arg1 glycoprotein 1589:1600 a model N-glycosylated glycoprotein 1566:1600 a model N-glycosylated glycoprotein 1566:1600 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 6 50 theme N-glycans 1097:1105 arg1 residues 1072:1079 the terminal α-(1,2)-linked mannose residues 1036:1079 the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans 1036:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 50 theme N-glycans 1097:1105 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 3 51 from protein 668:674 arg1 release 637:643 the release 633:643 the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues 633:763 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 3 51 from protein 668:674 arg1 glycan 652:657 the glycan 648:657 the glycan from the protein 648:674 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 2 52 theme abundant 299:306 arg1 form 308:311 an abundant form 296:311 an abundant form of protein glycosylation that has critical functional roles in the host 296:383 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 52 theme abundant 299:306 arg1 N-glycosylation 274:288 N-glycosylation 274:288 N-glycosylation 274:288 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 12 53 theme N-glycan 1992:1999 arg1 processing 2001:2010 N-glycan processing 1992:2010 N-glycan processing by S. pneumoniae 1992:2027 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 1 54 theme first 195:199 arg1 point 201:205 a first point 193:205 a first point of contact for colonizing and invading bacteria 193:253 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 9 55 from contribution 1611:1622 arg1 model 1676:1680 a mouse model 1668:1680 a mouse model 1668:1680 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 12 56 theme high-mannose 2055:2066 arg1 N-glycans 2068:2076 both complex and high-mannose N-glycans 2038:2076 N-glycans 2068:2076 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 2 57 theme glycosylation 324:336 arg1 form 308:311 an abundant form 296:311 an abundant form of protein glycosylation that has critical functional roles in the host 296:383 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 57 theme glycosylation 324:336 arg1 N-glycosylation 274:288 N-glycosylation 274:288 N-glycosylation 274:288 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 9 58 theme enzymes 1633:1639 arg1 SpGH92 1645:1650 SpGH92 1645:1650 SpGH92 to virulence in a mouse model 1645:1680 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 9 58 theme enzymes 1633:1639 arg1 contribution 1611:1622 the contribution 1607:1622 the contribution of these enzymes 1607:1639 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 0 59 from pneumoniae 82:91 arg1 Characterization 10:25 Molecular Characterization 0:25 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 0:126 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 11 60 theme glycan 1879:1884 arg1 transport 1853:1861 transport 1853:1861 transport of the released glycan 1853:1884 Therefore, N-glycan processing, but not transport of the released glycan, is required for full virulence in S. pneumoniae. 28056108 5 61 contain possesses 957:965 arg2 1,2 973:975 1,2 973:975 1,2 973:975 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 61 contain possesses 957:965 arg2 SpGH92 991:996 SpGH92 991:996 SpGH92 991:996 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 61 contain possesses 957:965 arg1 pneumoniae 941:950 S. pneumoniae 938:950 S. pneumoniae 938:950 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 5 61 contain possesses 957:965 arg2 -mannosidase 977:988 an α-(1,2)-mannosidase 967:988 an α-(1,2)-mannosidase (SpGH92) 967:997 Through biochemical and structural analyses we demonstrate that S. pneumoniae also possesses an α-(1,2)-mannosidase (SpGH92). 28056108 8 62 theme Binding 1285:1291 arg1 studies 1293:1299 Binding studies 1285:1299 Binding studies 1285:1299 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 2 63 theme functional 356:365 arg1 roles 367:371 critical functional roles 347:371 critical functional roles 347:371 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 1 64 theme human 161:165 arg1 tissues 167:173 human tissues 161:173 human tissues 161:173 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 8 65 theme activity 1448:1455 arg1 Man5GlcNAc 1417:1426 Man5GlcNAc 1417:1426 Man5GlcNAc 1417:1426 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 65 theme activity 1448:1455 arg1 product 1431:1437 a product 1429:1437 a product of EndoD activity 1429:1455 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 0 66 from Characterization 10:25 arg1 pneumoniae 82:91 Streptococcus pneumoniae 68:91 Streptococcus pneumoniae 68:91 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 2 67 theme protein 316:322 arg1 glycosylation 324:336 protein glycosylation 316:336 protein glycosylation 316:336 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 1 68 theme contact 210:216 arg1 point 201:205 a first point 193:205 a first point of contact for colonizing and invading bacteria 193:253 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 6 69 theme high-mannose 1084:1095 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 12 70 theme degradation 2126:2136 arg1 pathway 2138:2144 this degradation pathway 2121:2144 this degradation pathway 2121:2144 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 4 71 theme pneumoniae 801:810 arg1 capacity 786:793 the capacity 782:793 the capacity of S. pneumoniae to process high-mannose N-glycans and transport the products 782:871 Here we examine the capacity of S. pneumoniae to process high-mannose N-glycans and transport the products. 28056108 3 72 theme human 468:472 arg1 pneumoniae 497:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 8 73 theme high 1463:1466 arg1 affinity 1468:1475 high affinity 1463:1475 high affinity 1463:1475 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 74 theme solute 1347:1352 arg1 protein 1362:1368 the solute binding protein 1343:1368 the solute binding protein of an ABC transporter (ABCNG) 1343:1398 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 74 theme solute 1347:1352 arg1 NgtS 1337:1340 NgtS 1337:1340 NgtS 1337:1340 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 2 75 theme host-adapted 391:402 arg1 bacteria 404:411 some host-adapted bacteria 386:411 some host-adapted bacteria 386:411 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 0 76 theme N-glycan 30:37 arg1 Degradation 39:49 N-glycan Degradation 30:49 N-glycan Degradation 30:49 Molecular Characterization of N-glycan Degradation and Transport in Streptococcus pneumoniae and Its Contribution to Virulence. 28056108 9 77 theme EndoD 1520:1524 arg1 contribution 1504:1515 the contribution 1500:1515 the contribution of EndoD and ABCNG to growth of S. pneumoniae 1500:1561 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 6 78 theme -linked 1056:1062 arg1 residues 1072:1079 the terminal α-(1,2)-linked mannose residues 1036:1079 the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans 1036:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 6 78 theme -linked 1056:1062 arg1 N-glycans 1097:1105 high-mannose N-glycans 1084:1105 high-mannose N-glycans 1084:1105 This enzyme has the ability to trim the terminal α-(1,2)-linked mannose residues of high-mannose N-glycans to generate Man5GlcNAc2. 28056108 3 79 from release 637:643 arg1 protein 668:674 the protein 664:674 the protein 664:674 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 9 80 theme ABCNG 1530:1534 arg1 contribution 1504:1515 the contribution 1500:1515 the contribution of EndoD and ABCNG to growth of S. pneumoniae 1500:1561 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 12 81 theme complex 2043:2049 arg1 N-glycans 2068:2076 both complex and high-mannose N-glycans 2038:2076 N-glycans 2068:2076 To conclude, we synthesize our findings into a model of N-glycan processing by S. pneumoniae in which both complex and high-mannose N-glycans are targeted, and in which the two arms of this degradation pathway converge at ABCNG. 28056108 3 82 theme Streptococcus 483:495 arg1 pneumoniae 497:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae 464:506 The human pathogen Streptococcus pneumoniae depolymerizes complex N-glycans with enzymes that sequentially trim a complex N-glycan down to the Man3GlcNAc2 core prior to the release of the glycan from the protein by endo-β-N-acetylglucosaminidase (EndoD), which cleaves between the two GlcNAc residues. 28056108 7 83 theme -linked 1259:1265 arg1 residues 1275:1282 α-(1,2)-linked mannose residues 1252:1282 α-(1,2)-linked mannose residues 1252:1282 Through this activity SpGH92 is able to produce a substrate for EndoD, which is not active on high-mannose glycans with α-(1,2)-linked mannose residues. 28056108 8 84 theme transporter 1380:1390 arg1 protein 1362:1368 the solute binding protein 1343:1368 the solute binding protein of an ABC transporter (ABCNG) 1343:1398 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 8 84 theme transporter 1380:1390 arg1 NgtS 1337:1340 NgtS 1337:1340 NgtS 1337:1340 Binding studies and X-ray crystallography show that NgtS, the solute binding protein of an ABC transporter (ABCNG), is able to bind Man5GlcNAc, a product of EndoD activity, with high affinity. 28056108 9 85 gly N-glycosylated 1574:1587 arg1 glycoprotein 1589:1600 a model N-glycosylated glycoprotein 1566:1600 a model N-glycosylated glycoprotein 1566:1600 Finally, we evaluated the contribution of EndoD and ABCNG to growth of S. pneumoniae on a model N-glycosylated glycoprotein, and the contribution of these enzymes and SpGH92 to virulence in a mouse model. 28056108 2 86 contain has 343:345 arg1 form 308:311 an abundant form 296:311 an abundant form of protein glycosylation that has critical functional roles in the host 296:383 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 86 contain has 343:345 arg1 N-glycosylation 274:288 N-glycosylation 274:288 N-glycosylation 274:288 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 86 contain has 343:345 arg2 roles 367:371 critical functional roles 347:371 critical functional roles 347:371 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 87 link N-linked 446:453 arg1 glycans 455:461 N-linked glycans 446:461 N-linked glycans 446:461 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 2 88 theme critical 347:354 arg1 roles 367:371 critical functional roles 347:371 critical functional roles 347:371 Commensurate with N-glycosylation being an abundant form of protein glycosylation that has critical functional roles in the host, some host-adapted bacteria possess the machinery to process N-linked glycans. 28056108 1 89 theme invading 237:244 arg1 bacteria 246:253 colonizing and invading bacteria 222:253 colonizing and invading bacteria 222:253 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28056108 1 90 theme colonizing 222:231 arg1 bacteria 246:253 colonizing and invading bacteria 222:253 colonizing and invading bacteria 222:253 The carbohydrate-rich coating of human tissues and cells provide a first point of contact for colonizing and invading bacteria. 28510654 8 0 with incubation 965:974 arg1 substrate 1001:1009 a synthetic peptide substrate 981:1009 a synthetic peptide substrate 981:1009 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 8 1 theme oligosaccharide 1060:1074 arg1 moieties 1076:1083 isotopically labeled oligosaccharide moieties 1039:1083 isotopically labeled oligosaccharide moieties 1039:1083 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 7 2 theme donors 954:959 arg1 labeling 924:931 the metabolic labeling 910:931 the metabolic labeling of N-oligosaccharide donors 910:959 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 10 3 theme core 1352:1355 arg1 -β-GlcNAc-Asn 1420:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 3 theme core 1352:1355 arg1 α-Man- 1381:1386 α-Man-(1-6) 1381:1391 α-Man-(1-6) 1381:1391 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 3 theme core 1352:1355 arg1 α-Man- 1368:1373 α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1368:1432 α-Man- 1368:1373 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 3 theme core 1352:1355 arg1 structure 1357:1365 the eukaryotic core structure 1337:1365 the eukaryotic core structure 1337:1365 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 12 4 dep α-Man- 1782:1787 arg1 α-Man- 1795:1800 α-Man-(1-6)- 1795:1806 α-Man-(1-6)- 1795:1806 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 12 4 dep α-Man- 1782:1787 arg1 -β-GlcNAc3NAc-Asn 1839:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 0 5 link Asn-linked 0:9 arg1 chain 27:31 Asn-linked oligosaccharide chain 0:31 Asn-linked oligosaccharide chain of a crenarchaeon 0:49 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 0 5 link Asn-linked 0:9 arg1 calidifontis 64:75 Pyrobaculum calidifontis 52:75 Pyrobaculum calidifontis 52:75 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 11 6 dep modified 1651:1658 arg1 both 1646:1649 both 1646:1649 both 1646:1649 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 2 7 theme N-glycan 286:293 arg1 structure 295:303 only the N-glycan structure 277:303 only the N-glycan structure of the genus Sulfolobus 277:327 In contrast to the phylum Euryarchaeota, only the N-glycan structure of the genus Sulfolobus is known in Crenarchaeota. 28510654 7 8 theme metabolic 914:922 arg1 labeling 924:931 the metabolic labeling 910:931 the metabolic labeling of N-oligosaccharide donors 910:959 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 9 9 dep biantennary 1181:1191 arg1 high-mannose-type 1194:1210 high-mannose-type 1194:1210 high-mannose-type 1194:1210 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 0 10 theme glycan 129:134 arg1 reminiscent 81:91 reminiscent 81:91 reminiscent of the eukaryotic high-mannose-type glycan 81:134 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 2 11 theme genus 312:316 arg1 Sulfolobus 318:327 the genus Sulfolobus 308:327 the genus Sulfolobus 308:327 In contrast to the phylum Euryarchaeota, only the N-glycan structure of the genus Sulfolobus is known in Crenarchaeota. 28510654 4 12 theme MASCOT 460:465 arg1 search 467:472 The MASCOT search 456:472 The MASCOT search 456:472 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 9 13 theme magnetic 1105:1112 arg1 resonance 1114:1122 nuclear magnetic resonance 1097:1122 nuclear magnetic resonance 1097:1122 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 12 14 theme P. 1755:1756 arg1 calidifontisN-glycan 1758:1777 the P. calidifontisN-glycan 1751:1777 the P. calidifontisN-glycan 1751:1777 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 9 15 contain has 1175:1177 arg1 calidifontisN-glycan 1154:1173 the P. calidifontisN-glycan 1147:1173 the P. calidifontisN-glycan 1147:1173 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 9 15 contain has 1175:1177 arg2 structure 1212:1220 a biantennary, high-mannose-type structure 1179:1220 a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues 1179:1267 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 9 16 theme P. 1151:1152 arg1 calidifontisN-glycan 1154:1173 the P. calidifontisN-glycan 1147:1173 the P. calidifontisN-glycan 1147:1173 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 3 17 from cells 418:422 arg1 glycoproteins 374:386 glycoproteins 374:386 glycoproteins from cultured P. calidifontis cells 374:422 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 5 18 theme mass 555:558 arg1 MS/MS 574:578 MS/MS 574:578 MS/MS 574:578 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 5 18 theme mass 555:558 arg1 spectrometry 560:571 tandem mass spectrometry 548:571 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides 544:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 12 19 theme core 1733:1736 arg1 structure 1738:1746 the core structure 1729:1746 the core structure of the P. calidifontisN-glycan 1729:1777 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 12 19 theme core 1733:1736 arg1 α-Man- 1782:1787 α-Man-(1-3)- 1782:1793 α-Man-(1-3)- 1782:1793 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 4 20 theme potential 512:520 arg1 site 538:541 at least one potential N-glycosylation site 499:541 at least one potential N-glycosylation site 499:541 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 5 21 theme glycopeptides 610:622 arg1 analysis 581:588 The tandem mass spectrometry (MS/MS) analysis 544:588 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides 544:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 3 22 dep P. 402:403 arg1 calidifontis 405:416 calidifontis 405:416 calidifontis 405:416 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 9 23 theme biantennary 1181:1191 arg1 structure 1212:1220 a biantennary, high-mannose-type structure 1179:1220 a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues 1179:1267 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 12 24 theme calidifontisN-glycan 1897:1916 arg1 features 1878:1885 the unique features 1867:1885 the unique features of the P. calidifontisN-glycan 1867:1916 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 10 25 theme β-Man- 1394:1399 arg1 -β-GlcNAc-Asn 1420:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 25 theme β-Man- 1394:1399 arg1 structure 1357:1365 the eukaryotic core structure 1337:1365 the eukaryotic core structure 1337:1365 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 4 26 with proteins 485:492 arg1 site 538:541 at least one potential N-glycosylation site 499:541 at least one potential N-glycosylation site 499:541 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 0 27 theme eukaryotic 100:109 arg1 glycan 129:134 the eukaryotic high-mannose-type glycan 96:134 the eukaryotic high-mannose-type glycan 96:134 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 3 28 theme P. 402:403 arg1 cells 418:422 cultured P. calidifontis cells 393:422 cultured P. calidifontis cells 393:422 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 3 29 theme lectin 433:438 arg1 chromatography 440:453 ConA lectin chromatography 428:453 ConA lectin chromatography 428:453 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 0 30 theme Asn-linked 0:9 arg1 chain 27:31 Asn-linked oligosaccharide chain 0:31 Asn-linked oligosaccharide chain of a crenarchaeon 0:49 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 0 30 theme Asn-linked 0:9 arg1 calidifontis 64:75 Pyrobaculum calidifontis 52:75 Pyrobaculum calidifontis 52:75 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 5 31 theme small 596:600 arg1 glycopeptides 610:622 12 small tryptic glycopeptides 593:622 12 small tryptic glycopeptides 593:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 13 32 theme eukaryotic 2037:2046 arg1 counterparts 2048:2059 the eukaryotic counterparts 2033:2059 the eukaryotic counterparts 2033:2059 In spite of these differences, the structure of the P. calidifontisN-glycan is the most similar to the eukaryotic counterparts, among all archaeal N-glycans reported to date. 28510654 6 33 theme oligosaccharide 721:735 arg1 structure 737:745 the N-linked oligosaccharide structure 708:745 the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction 708:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 7 34 theme rich 863:866 arg1 medium 868:873 rich medium 863:873 rich medium supplemented with 13C-glucose 863:903 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 8 35 theme peptide 993:999 arg1 substrate 1001:1009 a synthetic peptide substrate 981:1009 a synthetic peptide substrate 981:1009 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 7 36 theme N-oligosaccharide 936:952 arg1 donors 954:959 N-oligosaccharide donors 936:959 N-oligosaccharide donors 936:959 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 11 37 theme chitobiose 1526:1535 arg1 structure 1537:1545 the chitobiose structure 1522:1545 the chitobiose structure 1522:1545 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 11 38 from position 1706:1713 arg1 group 1689:1693 an additional acetamido group 1665:1693 an additional acetamido group at the C-3 position 1665:1713 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 13 39 theme calidifontisN-glycan 1989:2008 arg1 structure 1969:1977 the structure 1965:1977 the structure of the P. calidifontisN-glycan 1965:2008 In spite of these differences, the structure of the P. calidifontisN-glycan is the most similar to the eukaryotic counterparts, among all archaeal N-glycans reported to date. 28510654 13 39 theme calidifontisN-glycan 1989:2008 arg1 similar 2022:2028 similar 2022:2028 similar 2022:2028 In spite of these differences, the structure of the P. calidifontisN-glycan is the most similar to the eukaryotic counterparts, among all archaeal N-glycans reported to date. 28510654 12 40 theme -β-GlcANAc3NAc- 1819:1833 arg1 -β-GlcNAc3NAc-Asn 1839:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 7 41 theme Pyrobaculum 815:825 arg1 cells 840:844 Pyrobaculum calidifontis cells 815:844 Pyrobaculum calidifontis cells 815:844 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 13 42 theme P. 1986:1987 arg1 calidifontisN-glycan 1989:2008 the P. calidifontisN-glycan 1982:2008 the P. calidifontisN-glycan 1982:2008 In spite of these differences, the structure of the P. calidifontisN-glycan is the most similar to the eukaryotic counterparts, among all archaeal N-glycans reported to date. 28510654 8 43 theme labeled 1052:1058 arg1 moieties 1076:1083 isotopically labeled oligosaccharide moieties 1039:1083 isotopically labeled oligosaccharide moieties 1039:1083 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 5 44 gly glycopeptides 610:622 arg2 glycopeptides 610:622 12 small tryptic glycopeptides 593:622 12 small tryptic glycopeptides 593:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 6 45 theme oligosaccharyl 781:794 arg1 reaction 805:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 10 46 theme calidifontisN-glycan 1297:1316 arg1 portion 1279:1285 The base portion 1270:1285 The base portion of the P. calidifontisN-glycan 1270:1316 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 8 47 gly glycopeptides 1020:1032 arg2 glycopeptides 1020:1032 glycopeptides 1020:1032 glycopeptides with isotopically labeled oligosaccharide moieties 1020:1083 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 11 48 theme acetamido 1679:1687 arg1 group 1689:1693 an additional acetamido group 1665:1693 an additional acetamido group at the C-3 position 1665:1713 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 11 49 theme C-3 1702:1704 arg1 position 1706:1713 the C-3 position 1698:1713 the C-3 position 1698:1713 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 10 50 theme eukaryotic 1341:1350 arg1 -β-GlcNAc-Asn 1420:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 50 theme eukaryotic 1341:1350 arg1 α-Man- 1381:1386 α-Man-(1-6) 1381:1391 α-Man-(1-6) 1381:1391 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 50 theme eukaryotic 1341:1350 arg1 α-Man- 1368:1373 α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1368:1432 α-Man- 1368:1373 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 50 theme eukaryotic 1341:1350 arg1 structure 1357:1365 the eukaryotic core structure 1337:1365 the eukaryotic core structure 1337:1365 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 11 51 theme anomeric 1471:1478 arg1 configuration 1480:1492 the anomeric configuration 1467:1492 the anomeric configuration between Man and GlcNAc 1467:1515 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 5 52 theme N-glycosylation 648:662 arg1 consensus 664:672 the canonical N-glycosylation consensus 634:672 the canonical N-glycosylation consensus 634:672 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 4 53 gly N-glycosylation 522:536 arg2 site 538:541 at least one potential N-glycosylation site 499:541 at least one potential N-glycosylation site 499:541 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 4 53 gly N-glycosylation 522:536 arg2 one 508:510 one 508:510 one 508:510 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 9 54 theme MS 1090:1091 arg1 analyses 1124:1131 The MS and nuclear magnetic resonance analyses 1086:1131 The MS and nuclear magnetic resonance analyses 1086:1131 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 11 55 theme GlcNAc 1626:1631 arg1 residues 1633:1640 the GlcNAc residues 1622:1640 the GlcNAc residues 1622:1640 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 2 56 theme Sulfolobus 318:327 arg1 structure 295:303 only the N-glycan structure 277:303 only the N-glycan structure of the genus Sulfolobus 277:327 In contrast to the phylum Euryarchaeota, only the N-glycan structure of the genus Sulfolobus is known in Crenarchaeota. 28510654 1 57 theme phylum 214:219 arg1 Crenarchaeota 221:233 the phylum Crenarchaeota 210:233 the phylum Crenarchaeota 210:233 Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. 28510654 9 58 theme nuclear 1097:1103 arg1 resonance 1114:1122 nuclear magnetic resonance 1097:1122 nuclear magnetic resonance 1097:1122 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 2 59 theme phylum 255:260 arg1 Euryarchaeota 262:274 the phylum Euryarchaeota 251:274 the phylum Euryarchaeota 251:274 In contrast to the phylum Euryarchaeota, only the N-glycan structure of the genus Sulfolobus is known in Crenarchaeota. 28510654 9 60 theme resonance 1114:1122 arg1 analyses 1124:1131 The MS and nuclear magnetic resonance analyses 1086:1131 The MS and nuclear magnetic resonance analyses 1086:1131 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 13 61 theme archaeal 2072:2079 arg1 N-glycans 2081:2089 all archaeal N-glycans 2068:2089 all archaeal N-glycans reported to date 2068:2106 In spite of these differences, the structure of the P. calidifontisN-glycan is the most similar to the eukaryotic counterparts, among all archaeal N-glycans reported to date. 28510654 11 62 mod modified 1651:1658 arg1 residues 1633:1640 the GlcNAc residues 1622:1640 the GlcNAc residues 1622:1640 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 11 62 mod modified 1651:1658 arg3 group 1689:1693 an additional acetamido group 1665:1693 an additional acetamido group at the C-3 position 1665:1713 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 1 63 theme Pyrobaculum 137:147 arg1 calidifontis 149:160 Pyrobaculum calidifontis 137:160 Pyrobaculum calidifontis 137:160 Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. 28510654 1 63 theme Pyrobaculum 137:147 arg1 archaeon 185:192 a hyperthermophilic archaeon 165:192 a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota 165:233 Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. 28510654 5 64 theme tandem 548:553 arg1 MS/MS 574:578 MS/MS 574:578 MS/MS 574:578 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 5 64 theme tandem 548:553 arg1 spectrometry 560:571 tandem mass spectrometry 548:571 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides 544:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 12 65 theme calidifontisN-glycan 1758:1777 arg1 structure 1738:1746 the core structure 1729:1746 the core structure of the P. calidifontisN-glycan 1729:1777 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 12 65 theme calidifontisN-glycan 1758:1777 arg1 α-Man- 1782:1787 α-Man-(1-3)- 1782:1793 α-Man-(1-3)- 1782:1793 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 4 66 theme N-glycosylation 522:536 arg1 site 538:541 at least one potential N-glycosylation site 499:541 at least one potential N-glycosylation site 499:541 The MASCOT search identified proteins with at least one potential N-glycosylation site. 28510654 12 67 theme P. 1894:1895 arg1 calidifontisN-glycan 1897:1916 the P. calidifontisN-glycan 1890:1916 the P. calidifontisN-glycan 1890:1916 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 5 68 theme spectrometry 560:571 arg1 analysis 581:588 The tandem mass spectrometry (MS/MS) analysis 544:588 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides 544:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 1 69 theme hyperthermophilic 167:183 arg1 calidifontis 149:160 Pyrobaculum calidifontis 137:160 Pyrobaculum calidifontis 137:160 Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. 28510654 1 69 theme hyperthermophilic 167:183 arg1 archaeon 185:192 a hyperthermophilic archaeon 165:192 a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota 165:233 Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. 28510654 11 70 theme GlcNAc 1575:1580 arg1 residue 1582:1588 one GlcNAc residue 1571:1588 one GlcNAc residue 1571:1588 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 6 71 theme in 762:763 arg1 reaction 805:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 0 72 theme high-mannose-type 111:127 arg1 glycan 129:134 the eukaryotic high-mannose-type glycan 96:134 the eukaryotic high-mannose-type glycan 96:134 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 6 73 link N-linked 712:719 arg1 structure 737:745 the N-linked oligosaccharide structure 708:745 the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction 708:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 5 74 theme canonical 638:646 arg1 consensus 664:672 the canonical N-glycosylation consensus 634:672 the canonical N-glycosylation consensus 634:672 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 0 75 theme oligosaccharide 11:25 arg1 chain 27:31 Asn-linked oligosaccharide chain 0:31 Asn-linked oligosaccharide chain of a crenarchaeon 0:49 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 0 75 theme oligosaccharide 11:25 arg1 calidifontis 64:75 Pyrobaculum calidifontis 52:75 Pyrobaculum calidifontis 52:75 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 8 76 with glycopeptides 1020:1032 arg1 moieties 1076:1083 isotopically labeled oligosaccharide moieties 1039:1083 isotopically labeled oligosaccharide moieties 1039:1083 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 6 77 dep in 762:763 arg1 vitro 765:769 vitro 765:769 vitro 765:769 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 12 78 theme unique 1871:1876 arg1 features 1878:1885 the unique features 1867:1885 the unique features of the P. calidifontisN-glycan 1867:1916 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 3 79 gly glycoproteins 374:386 arg1 glycoproteins 374:386 glycoproteins 374:386 glycoproteins from cultured P. calidifontis cells 374:422 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 5 80 theme tryptic 602:608 arg1 glycopeptides 610:622 12 small tryptic glycopeptides 593:622 12 small tryptic glycopeptides 593:622 The tandem mass spectrometry (MS/MS) analysis of 12 small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P. calidifontis. 28510654 3 81 theme cultured 393:400 arg1 cells 418:422 cultured P. calidifontis cells 393:422 cultured P. calidifontis cells 393:422 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 6 82 theme N-linked 712:719 arg1 structure 737:745 the N-linked oligosaccharide structure 708:745 the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction 708:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 0 83 theme crenarchaeon 38:49 arg1 chain 27:31 Asn-linked oligosaccharide chain 0:31 Asn-linked oligosaccharide chain of a crenarchaeon 0:49 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 0 83 theme crenarchaeon 38:49 arg1 calidifontis 64:75 Pyrobaculum calidifontis 52:75 Pyrobaculum calidifontis 52:75 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 8 84 theme synthetic 983:991 arg1 substrate 1001:1009 a synthetic peptide substrate 981:1009 a synthetic peptide substrate 981:1009 An incubation with a synthetic peptide substrate produced glycopeptides with isotopically labeled oligosaccharide moieties. 28510654 3 85 theme ConA 428:431 arg1 chromatography 440:453 ConA lectin chromatography 428:453 ConA lectin chromatography 428:453 Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. 28510654 11 86 theme Structural 1435:1444 arg1 differences 1446:1456 Structural differences 1435:1456 Structural differences 1435:1456 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 9 87 theme monosaccharide 1245:1258 arg1 residues 1260:1267 up to 11 monosaccharide residues 1236:1267 up to 11 monosaccharide residues 1236:1267 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 0 88 theme Pyrobaculum 52:62 arg1 chain 27:31 Asn-linked oligosaccharide chain 0:31 Asn-linked oligosaccharide chain of a crenarchaeon 0:49 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 0 88 theme Pyrobaculum 52:62 arg1 calidifontis 64:75 Pyrobaculum calidifontis 52:75 Pyrobaculum calidifontis 52:75 Asn-linked oligosaccharide chain of a crenarchaeon, Pyrobaculum calidifontis, is reminiscent of the eukaryotic high-mannose-type glycan. 28510654 12 89 theme α-Man- 1808:1813 arg1 -β-GlcNAc3NAc-Asn 1839:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn 1808:1855 As a result, the core structure of the P. calidifontisN-glycan is α-Man-(1-3)-(α-Man-(1-6)-)α-Man-(1-4)-β-GlcANAc3NAc-(1-4)-β-GlcNAc3NAc-Asn, in which the unique features of the P. calidifontisN-glycan are underlined. 28510654 10 90 theme -β-GlcNAc- 1405:1414 arg1 -β-GlcNAc-Asn 1420:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 )β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn 1393:1432 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 90 theme -β-GlcNAc- 1405:1414 arg1 structure 1357:1365 the eukaryotic core structure 1337:1365 the eukaryotic core structure 1337:1365 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 10 91 theme base 1274:1277 arg1 portion 1279:1285 The base portion 1270:1285 The base portion of the P. calidifontisN-glycan 1270:1316 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 28510654 6 92 theme enzymatic 771:779 arg1 reaction 805:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 6 93 theme transfer 796:803 arg1 reaction 805:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 an in vitro enzymatic oligosaccharyl transfer reaction 759:812 We determined the N-linked oligosaccharide structure produced by an in vitro enzymatic oligosaccharyl transfer reaction. 28510654 7 94 theme calidifontis 827:838 arg1 cells 840:844 Pyrobaculum calidifontis cells 815:844 Pyrobaculum calidifontis cells 815:844 Pyrobaculum calidifontis cells were cultured in rich medium supplemented with 13C-glucose, for the metabolic labeling of N-oligosaccharide donors. 28510654 9 95 dep 11 1242:1243 arg1 to 1239:1240 to 1239:1240 to 1239:1240 The MS and nuclear magnetic resonance analyses revealed that the P. calidifontisN-glycan has a biantennary, high-mannose-type structure consisting of up to 11 monosaccharide residues. 28510654 11 96 theme additional 1668:1677 arg1 group 1689:1693 an additional acetamido group 1665:1693 an additional acetamido group at the C-3 position 1665:1713 Structural differences exist in the anomeric configuration between Man and GlcNAc, and the chitobiose structure is chemically modified: one GlcNAc residue is oxidized to glucoronate, and the GlcNAc residues are both modified with an additional acetamido group at the C-3 position. 28510654 10 97 theme P. 1294:1295 arg1 calidifontisN-glycan 1297:1316 the P. calidifontisN-glycan 1290:1316 the P. calidifontisN-glycan 1290:1316 The base portion of the P. calidifontisN-glycan strongly resembles the eukaryotic core structure, α-Man-(1-3)-(α-Man-(1-6)-)β-Man-(1-4)-β-GlcNAc-(1-4)-β-GlcNAc-Asn. 25850302 0 0 from thermocellum 176:187 arg1 glucuronoxylan-xylanohydrolase 78:107 first glucuronoxylan-xylanohydrolase 72:107 first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum] 72:188 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 0 from thermocellum 176:187 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 0 from thermocellum 176:187 arg1 Xyn30a 110:115 Xyn30a 110:115 Xyn30a 110:115 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 0 from thermocellum 176:187 arg1 studies 61:67 molecular docking studies 43:67 molecular docking studies 43:67 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 9 1 theme Multiple 1170:1177 arg1 alignment 1188:1196 Multiple sequence alignment 1170:1196 Multiple sequence alignment 1170:1196 Multiple sequence alignment showed that these catalytic residues are conserved within the family. 25850302 3 2 theme XynC 526:529 arg1 template 553:560 a template 551:560 a template to generate the molecular model 551:592 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 3 2 theme XynC 526:529 arg1 structure 513:521 crystal structure 505:521 crystal structure of XynC from B. subtilis 505:546 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 8 3 theme catalytic 1147:1155 arg1 nucleophile 1157:1167 a catalytic nucleophile 1145:1167 a catalytic nucleophile 1145:1167 Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. 25850302 8 3 theme catalytic 1147:1155 arg1 Glu225 1131:1136 Glu225 1131:1136 Glu225 1131:1136 Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. 25850302 6 4 theme beta/alpha 905:914 arg1 fold 925:928 a (beta/alpha)8 barrel fold 902:928 a (beta/alpha)8 barrel fold 902:928 Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. 25850302 2 5 theme full 303:306 arg1 CtXynGH30 315:323 The full length CtXynGH30 299:323 The full length CtXynGH30 299:323 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 1 6 theme thermocellum 285:296 arg1 cellulosome 258:268 cellulosome 258:268 cellulosome of Clostridium thermocellum 258:296 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 11 7 theme aromatic 1441:1448 arg1 acids 1456:1460 aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 1441:1533 aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 1441:1533 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 12 8 theme glycosyl 1742:1749 arg1 GH30 1762:1765 GH30 1762:1765 GH30 1762:1765 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 8 theme glycosyl 1742:1749 arg1 hydrolase 1751:1759 family 30 glycosyl hydrolase 1732:1759 family 30 glycosyl hydrolase (GH30) 1732:1766 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 9 from residues 1646:1653 arg1 thermocellum 1785:1796 Clostridium thermocellum 1773:1796 Clostridium thermocellum 1773:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 9 10 theme catalytic 1216:1224 arg1 residues 1226:1233 these catalytic residues 1210:1233 these catalytic residues 1210:1233 Multiple sequence alignment showed that these catalytic residues are conserved within the family. 25850302 3 11 theme molecular 578:586 arg1 model 588:592 the molecular model 574:592 the molecular model 574:592 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 12 12 theme overall 1596:1602 arg1 feature 1615:1621 overall structural feature 1596:1621 overall structural feature 1596:1621 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 11 13 theme binding 1542:1548 arg1 pocket 1555:1560 binding site pocket 1542:1560 binding site pocket 1542:1560 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 0 14 theme glycosyl 133:140 arg1 GH30 153:156 GH30 153:156 GH30 153:156 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 14 theme glycosyl 133:140 arg1 hydrolase 142:150 family-30 glycosyl hydrolase 123:150 family-30 glycosyl hydrolase (GH30) 123:157 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 5 15 theme allowed 841:847 arg1 region 849:854 additional allowed region 830:854 additional allowed region 830:854 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 4 16 theme steepest 663:670 arg1 algorithm 680:688 steepest descent algorithm 663:688 steepest descent algorithm 663:688 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25850302 11 17 theme binding 1396:1402 arg1 interaction 1404:1414 The binding interaction 1392:1414 The binding interaction of ligands 1392:1425 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 5 18 theme Ramachandran 744:755 arg1 plot 757:760 Ramachandran plot 744:760 Ramachandran plot 744:760 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 1 19 theme modular 226:232 arg1 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 1 19 theme modular 226:232 arg1 enzyme 234:239 a carbohydrate active modular enzyme and component 204:253 enzyme 234:239 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 7 20 contain had 950:952 arg1 it 947:948 it 947:948 it 947:948 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 7 20 contain had 950:952 arg2 structure 973:981 a beta-strand rich structure 954:981 a beta-strand rich structure called 'side beta-structure' attached with main catalytic core 954:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 7 21 theme catalytic 1031:1039 arg1 core 1041:1044 main catalytic core 1026:1044 main catalytic core 1026:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 11 22 theme site 1550:1553 arg1 pocket 1555:1560 binding site pocket 1542:1560 binding site pocket 1542:1560 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 4 23 theme force 709:713 arg1 field 715:719 GROMOS96 43al force field 695:719 GROMOS96 43al force field 695:719 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25850302 5 24 theme amino 778:782 arg1 acids 784:788 91% amino acids 774:788 91% amino acids 774:788 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 7 25 theme beta-strand 956:966 arg1 structure 973:981 a beta-strand rich structure 954:981 a beta-strand rich structure called 'side beta-structure' attached with main catalytic core 954:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 12 26 theme glucuronoxylan-xylanohydrolase 1689:1718 arg1 docking 1672:1678 enzymeligand docking 1659:1678 enzymeligand docking 1659:1678 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 26 theme glucuronoxylan-xylanohydrolase 1689:1718 arg1 residues 1646:1653 conserved active site residues 1624:1653 conserved active site residues 1624:1653 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 26 theme glucuronoxylan-xylanohydrolase 1689:1718 arg1 feature 1615:1621 overall structural feature 1596:1621 overall structural feature 1596:1621 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 26 theme glucuronoxylan-xylanohydrolase 1689:1718 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 0 27 theme glucuronoxylan-xylanohydrolase 78:107 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 27 theme glucuronoxylan-xylanohydrolase 78:107 arg1 studies 61:67 molecular docking studies 43:67 molecular docking studies 43:67 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 7 28 dep ditionally 935:944 arg1 Ad- 931:933 Ad- 931:933 Ad- 931:933 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 5 29 from % 825:825 arg1 region 849:854 additional allowed region 830:854 additional allowed region 830:854 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 12 30 theme enzymeligand 1659:1670 arg1 docking 1672:1678 enzymeligand docking 1659:1678 enzymeligand docking 1659:1678 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 5 31 theme 91 774:775 arg1 % 776:776 % 776:776 % 776:776 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 0 32 theme first 72:76 arg1 glucuronoxylan-xylanohydrolase 78:107 first glucuronoxylan-xylanohydrolase 72:107 first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum] 72:188 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 32 theme first 72:76 arg1 Xyn30a 110:115 Xyn30a 110:115 Xyn30a 110:115 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 2 33 theme carbohydrate 396:407 arg1 CBM6 425:428 CBM6 425:428 CBM6 425:428 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 33 theme carbohydrate 396:407 arg1 module 417:422 a family 6 carbohydrate binding module 385:422 a family 6 carbohydrate binding module (CBM6) 385:429 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 0 34 theme [In 0:2 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 8 35 theme Structural 1047:1056 arg1 superimposition 1058:1072 Structural superimposition 1047:1072 Structural superimposition 1047:1072 Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. 25850302 12 36 theme active 1634:1639 arg1 residues 1646:1653 conserved active site residues 1624:1653 conserved active site residues 1624:1653 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 36 theme active 1634:1639 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 10 37 theme polar 1323:1327 arg1 interaction 1329:1339 polar interaction 1323:1339 polar interaction with linear and substituted xylo-oligosaccharides 1323:1389 The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. 25850302 3 38 theme computer 469:476 arg1 program 478:484 computer program Modeller9v8 469:496 computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model 469:592 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 0 39 theme structural 11:20 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 12 40 from feature 1615:1621 arg1 thermocellum 1785:1796 Clostridium thermocellum 1773:1796 Clostridium thermocellum 1773:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 3 41 theme crystal 505:511 arg1 template 553:560 a template 551:560 a template to generate the molecular model 551:592 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 3 41 theme crystal 505:511 arg1 structure 513:521 crystal structure 505:521 crystal structure of XynC from B. subtilis 505:546 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 0 42 theme docking 53:59 arg1 studies 61:67 molecular docking studies 43:67 molecular docking studies 43:67 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 8 43 theme catalytic 1105:1113 arg1 Glu136 1089:1094 Glu136 1089:1094 Glu136 1089:1094 Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. 25850302 8 43 theme catalytic 1105:1113 arg1 acid/base 1115:1123 a catalytic acid/base 1103:1123 a catalytic acid/base 1103:1123 Structural superimposition reflected that Glu136 act as a catalytic acid/base while Glu225 act as a catalytic nucleophile. 25850302 11 44 theme ligands 1419:1425 arg1 interaction 1404:1414 The binding interaction 1392:1414 The binding interaction of ligands 1392:1425 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 3 45 from subtilis 539:546 arg1 template 553:560 a template 551:560 a template to generate the molecular model 551:592 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 3 45 from subtilis 539:546 arg1 structure 513:521 crystal structure 505:521 crystal structure of XynC from B. subtilis 505:546 Xyn30A was modeled by computer program Modeller9v8 taking crystal structure of XynC from B. subtilis as a template to generate the molecular model. 25850302 2 46 theme family 387:392 arg1 CBM6 425:428 CBM6 425:428 CBM6 425:428 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 46 theme family 387:392 arg1 module 417:422 a family 6 carbohydrate binding module 385:422 a family 6 carbohydrate binding module (CBM6) 385:429 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 6 47 theme barrel 918:923 arg1 fold 925:928 a (beta/alpha)8 barrel fold 902:928 a (beta/alpha)8 barrel fold 902:928 Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. 25850302 1 48 theme cellulosome 258:268 arg1 component 245:253 a carbohydrate active modular enzyme and component 204:253 component 245:253 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 1 48 theme cellulosome 258:268 arg1 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 1 48 theme cellulosome 258:268 arg1 enzyme 234:239 a carbohydrate active modular enzyme and component 204:253 enzyme 234:239 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 11 49 theme amino 1450:1454 arg1 acids 1456:1460 aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 1441:1533 aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 1441:1533 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 9 50 theme sequence 1179:1186 arg1 alignment 1188:1196 Multiple sequence alignment 1170:1196 Multiple sequence alignment 1170:1196 Multiple sequence alignment showed that these catalytic residues are conserved within the family. 25850302 2 51 theme length 308:313 arg1 CtXynGH30 315:323 The full length CtXynGH30 299:323 The full length CtXynGH30 299:323 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 1 52 theme Clostridium 273:283 arg1 thermocellum 285:296 Clostridium thermocellum 273:296 Clostridium thermocellum 273:296 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 12 53 from docking 1672:1678 arg1 thermocellum 1785:1796 Clostridium thermocellum 1773:1796 Clostridium thermocellum 1773:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 4 54 theme Model 595:599 arg1 refinement 601:610 Model refinement 595:610 Model refinement 595:610 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25850302 12 55 theme hydrolase 1751:1759 arg1 Xyn30A 1721:1726 Xyn30A 1721:1726 Xyn30A 1721:1726 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 55 theme hydrolase 1751:1759 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 4 56 theme descent 672:678 arg1 algorithm 680:688 steepest descent algorithm 663:688 steepest descent algorithm 663:688 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25850302 11 57 theme active 1573:1578 arg1 site 1580:1583 the active site 1569:1583 the active site 1569:1583 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 2 58 theme N-terminal 337:346 arg1 module 358:363 an N-terminal catalytic module 334:363 an N-terminal catalytic module named as Xyn30A 334:379 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 12 59 theme structural 1604:1613 arg1 feature 1615:1621 overall structural feature 1596:1621 overall structural feature 1596:1621 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 5 60 theme Quality 722:728 arg1 assessment 730:739 Quality assessment 722:739 Quality assessment by Ramachandran plot 722:760 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 5 61 theme additional 830:839 arg1 region 849:854 additional allowed region 830:854 additional allowed region 830:854 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 6 62 contain has 898:900 arg2 fold 925:928 a (beta/alpha)8 barrel fold 902:928 a (beta/alpha)8 barrel fold 902:928 Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. 25850302 6 62 contain has 898:900 arg1 Xyn30A 891:896 Xyn30A 891:896 Xyn30A 891:896 Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. 25850302 1 63 theme active 219:224 arg1 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 1 63 theme active 219:224 arg1 enzyme 234:239 a carbohydrate active modular enzyme and component 204:253 enzyme 234:239 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 7 64 theme main 1026:1029 arg1 core 1041:1044 main catalytic core 1026:1044 main catalytic core 1026:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 7 65 theme side 991:994 arg1 beta-structure 996:1009 'side beta-structure' attached with main catalytic core 990:1044 'side beta-structure' attached with main catalytic core 990:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 11 66 dep acids 1456:1460 arg1 Trp81 1462:1466 Trp81 1462:1466 Trp81 1462:1466 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 Trp264 1517:1522 Trp264 1517:1522 Trp264 1517:1522 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 Tyr139 1469:1474 Tyr139 1469:1474 Tyr139 1469:1474 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 His198 1493:1498 His198 1493:1498 His198 1493:1498 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 Tyr265 1528:1533 Tyr265 1528:1533 Tyr265 1528:1533 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 Trp143 1477:1482 Trp143 1477:1482 Trp143 1477:1482 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 11 66 dep acids 1456:1460 arg1 Tyr227 1509:1514 Tyr227 1509:1514 Tyr227 1509:1514 The binding interaction of ligands depicted that aromatic amino acids Trp81, Tyr139, Trp143, Phe172, His198, Tyr200, Tyr227, Trp264 and Tyr265 create binding site pocket around the active site. 25850302 5 67 theme % 776:776 arg1 acids 784:788 91% amino acids 774:788 91% amino acids 774:788 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 12 68 theme family 1732:1737 arg1 GH30 1762:1765 GH30 1762:1765 GH30 1762:1765 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 68 theme family 1732:1737 arg1 hydrolase 1751:1759 family 30 glycosyl hydrolase 1732:1759 family 30 glycosyl hydrolase (GH30) 1732:1766 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 0 69 from characterization 22:37 arg1 GH30 153:156 GH30 153:156 GH30 153:156 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 69 from characterization 22:37 arg1 hydrolase 142:150 family-30 glycosyl hydrolase 123:150 family-30 glycosyl hydrolase (GH30) 123:157 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 69 from characterization 22:37 arg1 thermocellum 176:187 Clostridium thermocellum 164:187 Clostridium thermocellum 164:187 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 4 70 theme 43al 704:707 arg1 field 715:719 GROMOS96 43al force field 695:719 GROMOS96 43al force field 695:719 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25850302 10 71 theme docking 1272:1278 arg1 results 1280:1286 The docking results 1268:1286 The docking results 1268:1286 The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. 25850302 5 72 from region 813:818 arg1 region 849:854 additional allowed region 830:854 additional allowed region 830:854 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 7 73 theme rich 968:971 arg1 structure 973:981 a beta-strand rich structure 954:981 a beta-strand rich structure called 'side beta-structure' attached with main catalytic core 954:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 12 74 theme first 1683:1687 arg1 Xyn30A 1721:1726 Xyn30A 1721:1726 Xyn30A 1721:1726 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 74 theme first 1683:1687 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 0 75 from studies 61:67 arg1 GH30 153:156 GH30 153:156 GH30 153:156 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 75 from studies 61:67 arg1 hydrolase 142:150 family-30 glycosyl hydrolase 123:150 family-30 glycosyl hydrolase (GH30) 123:157 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 75 from studies 61:67 arg1 thermocellum 176:187 Clostridium thermocellum 164:187 Clostridium thermocellum 164:187 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 76 theme family-30 123:131 arg1 GH30 153:156 GH30 153:156 GH30 153:156 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 76 theme family-30 123:131 arg1 hydrolase 142:150 family-30 glycosyl hydrolase 123:150 family-30 glycosyl hydrolase (GH30) 123:157 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 1 77 theme carbohydrate 206:217 arg1 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 191:199 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 1 77 theme carbohydrate 206:217 arg1 enzyme 234:239 a carbohydrate active modular enzyme and component 204:253 enzyme 234:239 CtXynGH30 is a carbohydrate active modular enzyme and component of cellulosome of Clostridium thermocellum. 25850302 12 78 from thermocellum 1785:1796 arg1 docking 1672:1678 enzymeligand docking 1659:1678 enzymeligand docking 1659:1678 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 78 from thermocellum 1785:1796 arg1 residues 1646:1653 conserved active site residues 1624:1653 conserved active site residues 1624:1653 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 78 from thermocellum 1785:1796 arg1 feature 1615:1621 overall structural feature 1596:1621 overall structural feature 1596:1621 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 78 from thermocellum 1785:1796 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 78 from thermocellum 1785:1796 arg1 Xyn30A 1721:1726 Xyn30A 1721:1726 Xyn30A 1721:1726 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 7 79 dep beta-structure 996:1009 arg1 attached 1012:1019 attached 1012:1019 attached with main catalytic core 1012:1044 Ad- ditionally, it had a beta-strand rich structure called 'side beta-structure' attached with main catalytic core. 25850302 2 80 theme binding 409:415 arg1 CBM6 425:428 CBM6 425:428 CBM6 425:428 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 80 theme binding 409:415 arg1 module 417:422 a family 6 carbohydrate binding module 385:422 a family 6 carbohydrate binding module (CBM6) 385:429 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 0 81 theme silico 4:9 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 10 82 with interaction 1329:1339 arg1 linear 1346:1351 linear 1346:1351 linear 1346:1351 The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. 25850302 10 82 with interaction 1329:1339 arg1 xylo-oligosaccharides 1369:1389 substituted xylo-oligosaccharides 1357:1389 substituted xylo-oligosaccharides 1357:1389 The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. 25850302 12 83 theme conserved 1624:1632 arg1 residues 1646:1653 conserved active site residues 1624:1653 conserved active site residues 1624:1653 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 83 theme conserved 1624:1632 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 0 84 theme molecular 43:51 arg1 studies 61:67 molecular docking studies 43:67 molecular docking studies 43:67 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 12 85 theme site 1641:1644 arg1 residues 1646:1653 conserved active site residues 1624:1653 conserved active site residues 1624:1653 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 12 85 theme site 1641:1644 arg1 glucuronoxylan-xylanohydrolase 1689:1718 first glucuronoxylan-xylanohydrolase 1683:1718 first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum 1683:1796 We report overall structural feature, conserved active site residues and enzymeligand docking of first glucuronoxylan-xylanohydrolase (Xyn30A) of family 30 glycosyl hydrolase (GH30) from Clostridium thermocellum. 25850302 0 86 from hydrolase 142:150 arg1 glucuronoxylan-xylanohydrolase 78:107 first glucuronoxylan-xylanohydrolase 72:107 first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum] 72:188 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 86 from hydrolase 142:150 arg1 characterization 22:37 [In silico structural characterization 0:37 [In silico structural characterization 0:37 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 86 from hydrolase 142:150 arg1 Xyn30a 110:115 Xyn30a 110:115 Xyn30a 110:115 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 0 86 from hydrolase 142:150 arg1 studies 61:67 molecular docking studies 43:67 molecular docking studies 43:67 [In silico structural characterization and molecular docking studies of first glucuronoxylan-xylanohydrolase (Xyn30a) from family-30 glycosyl hydrolase (GH30) from Clostridium thermocellum]. 25850302 10 87 theme substituted 1357:1367 arg1 xylo-oligosaccharides 1369:1389 substituted xylo-oligosaccharides 1357:1389 substituted xylo-oligosaccharides 1357:1389 The docking results showed that these residues display polar interaction with linear and substituted xylo-oligosaccharides. 25850302 5 88 theme favourable 802:811 arg1 region 813:818 most favourable region 797:818 most favourable region 797:818 Quality assessment by Ramachandran plot showed that 91% amino acids lie in most favourable region and 9% in additional allowed region. 25850302 2 89 contain contains 325:332 arg1 C-terminuis 434:444 C-terminuis 434:444 C-terminuis 434:444 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 89 contain contains 325:332 arg2 module 358:363 an N-terminal catalytic module 334:363 an N-terminal catalytic module named as Xyn30A 334:379 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 89 contain contains 325:332 arg2 CtXynGH30 315:323 The full length CtXynGH30 299:323 The full length CtXynGH30 299:323 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 89 contain contains 325:332 arg2 module 417:422 a family 6 carbohydrate binding module 385:422 a family 6 carbohydrate binding module (CBM6) 385:429 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 89 contain contains 325:332 arg2 CBM6 425:428 CBM6 425:428 CBM6 425:428 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 2 89 contain contains 325:332 arg1 CtXynGH30 315:323 The full length CtXynGH30 299:323 The full length CtXynGH30 299:323 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 6 90 theme Structural 857:866 arg1 analysis 868:875 Structural analysis 857:875 Structural analysis 857:875 Structural analysis depicted that Xyn30A has a (beta/alpha)8 barrel fold. 25850302 2 91 theme catalytic 348:356 arg1 module 358:363 an N-terminal catalytic module 334:363 an N-terminal catalytic module named as Xyn30A 334:379 The full length CtXynGH30 contains an N-terminal catalytic module named as Xyn30A and a family 6 carbohydrate binding module (CBM6) at C-terminuis. 25850302 4 92 theme energy 627:632 arg1 minimization 634:645 energy minimization 627:645 energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field 627:719 Model refinement was done using energy minimization by implementing steepest descent algorithm with GROMOS96 43al force field. 25449758 4 0 theme Mass 593:596 arg1 analysis 612:619 Mass spectrometric analysis 593:619 Mass spectrometric analysis of the fluorescently labeled N-glycan 593:657 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 25449758 7 1 theme mass 1005:1008 arg1 spectrometry 1010:1021 glycopeptide mass spectrometry 992:1021 glycopeptide mass spectrometry 992:1021 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 6 2 theme eukaryotic 954:963 arg1 pathway 974:980 an early eukaryotic dolichol pathway 945:980 an early eukaryotic dolichol pathway 945:980 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 0 3 from Production 0:9 arg1 coli 93:96 Escherichia coli 81:96 Escherichia coli 81:96 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 7 4 theme engineered 1085:1094 arg1 motif 1110:1114 an engineered glycosylation motif 1082:1114 an engineered glycosylation motif of recombinant maltose binding protein 1082:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 6 5 theme identical 915:923 arg1 Man3-GlcNAc2 901:912 a Man3-GlcNAc2 899:912 a Man3-GlcNAc2 899:912 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 2 6 theme lipid-linked 341:352 arg1 Man-GlcNAc2 375:385 Man-GlcNAc2 375:385 Man-GlcNAc2 375:385 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 2 6 theme lipid-linked 341:352 arg1 mannosyl-chitobiose 354:372 lipid-linked mannosyl-chitobiose 341:372 lipid-linked mannosyl-chitobiose (Man-GlcNAc2) 341:386 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 1 7 from mechanism 171:179 arg1 eukaryotes 184:193 eukaryotes 184:193 eukaryotes 184:193 N-Glycosylation is a ubiquitous protein post-translational modification mechanism in eukaryotes. 25449758 3 8 theme Campylobacter 571:583 arg1 jejuni 585:590 Campylobacter jejuni 571:590 Campylobacter jejuni 571:590 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 8 9 theme Man3-GlcNAc2 1174:1185 arg1 structure 1187:1195 the Man3-GlcNAc2 structure 1170:1195 the Man3-GlcNAc2 structure but not Man-GlcNAc2 1170:1215 Surprisingly, the Man3-GlcNAc2 structure but not Man-GlcNAc2 was transferred onto maltose binding protein. 25449758 9 10 theme PglB 1285:1288 arg1 protein 1290:1296 PglB protein 1285:1296 PglB protein 1285:1296 This work showed that PglB protein might be able to accommodate the transfer of the further engineered glycan with greater complexity. 25449758 3 11 theme E. 516:517 arg1 cell 524:527 the engineered E. coli cell 501:527 the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni 501:590 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 7 12 theme protein 1147:1153 arg1 motif 1110:1114 an engineered glycosylation motif 1082:1114 an engineered glycosylation motif of recombinant maltose binding protein 1082:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 7 13 theme assembled 1053:1061 arg1 moiety 1070:1075 the assembled glycan moiety 1049:1075 the assembled glycan moiety 1049:1075 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 9 14 theme greater 1378:1384 arg1 complexity 1386:1395 greater complexity 1378:1395 greater complexity 1378:1395 This work showed that PglB protein might be able to accommodate the transfer of the further engineered glycan with greater complexity. 25449758 7 15 theme moiety 1070:1075 arg1 transfer 1037:1044 the transfer 1033:1044 the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein 1033:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 7 16 gly glycosylation 1096:1108 arg2 motif 1110:1114 an engineered glycosylation motif 1082:1114 an engineered glycosylation motif of recombinant maltose binding protein 1082:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 5 17 theme reversed-phase 730:743 arg1 analysis 750:757 The reversed-phase HPLC analysis 726:757 The reversed-phase HPLC analysis 726:757 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 7 18 theme glycan 1063:1068 arg1 moiety 1070:1075 the assembled glycan moiety 1049:1075 the assembled glycan moiety 1049:1075 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 3 19 theme model 463:467 arg1 protein 481:487 a model periplasmic protein 461:487 a model periplasmic protein 461:487 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 5 20 theme HPLC 745:748 arg1 analysis 750:757 The reversed-phase HPLC analysis 726:757 The reversed-phase HPLC analysis 726:757 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 3 21 theme engineered 505:514 arg1 cell 524:527 the engineered E. coli cell 501:527 the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni 501:590 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 3 22 theme Man3-GlcNAc2 443:454 arg1 Transfer 431:438 Transfer 431:438 Transfer of Man3-GlcNAc2 onto a model periplasmic protein 431:487 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 9 23 theme engineered 1355:1364 arg1 glycan 1366:1371 the further engineered glycan 1343:1371 the further engineered glycan 1343:1371 This work showed that PglB protein might be able to accommodate the transfer of the further engineered glycan with greater complexity. 25449758 3 24 theme oligosaccharyltransferase 535:559 arg1 PglB 561:564 oligosaccharyltransferase PglB 535:564 oligosaccharyltransferase PglB 535:564 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 0 25 theme initial-stage 14:26 arg1 N-glycan 39:46 initial-stage eukaryotic N-glycan 14:46 initial-stage eukaryotic N-glycan 14:46 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 2 26 link lipid-linked 341:352 arg1 Man-GlcNAc2 375:385 Man-GlcNAc2 375:385 Man-GlcNAc2 375:385 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 2 26 link lipid-linked 341:352 arg1 mannosyl-chitobiose 354:372 lipid-linked mannosyl-chitobiose 341:372 lipid-linked mannosyl-chitobiose (Man-GlcNAc2) 341:386 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 5 27 theme α1,6- 803:807 arg1 mannoses 825:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 5 27 theme α1,6- 803:807 arg1 residues 782:789 the Hex residues 774:789 the Hex residues 774:789 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 9 28 theme glycan 1366:1371 arg1 transfer 1331:1338 the transfer 1327:1338 the transfer of the further engineered glycan with greater complexity 1327:1395 This work showed that PglB protein might be able to accommodate the transfer of the further engineered glycan with greater complexity. 25449758 4 29 theme glycan 671:676 arg1 signal 678:683 a glycan signal 669:683 a glycan signal composed of 2 HexNAc and 3 Hex residues 669:723 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 25449758 8 30 theme maltose 1238:1244 arg1 protein 1254:1260 maltose binding protein 1238:1260 maltose binding protein 1238:1260 Surprisingly, the Man3-GlcNAc2 structure but not Man-GlcNAc2 was transferred onto maltose binding protein. 25449758 7 31 gly glycopeptide 992:1003 arg2 glycopeptide 992:1003 glycopeptide mass spectrometry 992:1021 glycopeptide mass spectrometry 992:1021 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 0 32 theme N-glycan 39:46 arg1 Production 0:9 Production 0:9 Production of initial-stage eukaryotic N-glycan 0:46 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 0 32 theme N-glycan 39:46 arg1 glycosylation 64:76 its protein glycosylation 52:76 its protein glycosylation in Escherichia coli 52:96 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 5 33 theme β1,4-linked 813:823 arg1 mannoses 825:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 5 33 theme β1,4-linked 813:823 arg1 residues 782:789 the Hex residues 774:789 the Hex residues 774:789 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 6 34 theme dolichol 965:972 arg1 pathway 974:980 an early eukaryotic dolichol pathway 945:980 an early eukaryotic dolichol pathway 945:980 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 0 35 theme eukaryotic 28:37 arg1 N-glycan 39:46 initial-stage eukaryotic N-glycan 14:46 initial-stage eukaryotic N-glycan 14:46 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 6 36 theme constructed 868:878 arg1 system 880:885 the constructed system 864:885 the constructed system 864:885 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 2 37 contain containing 230:239 arg2 glycosyltransferases 241:260 glycosyltransferases 241:260 glycosyltransferases 241:260 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 2 37 contain containing 230:239 arg1 pathway 222:228 a synthetic pathway 210:228 a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae 210:290 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 0 38 theme protein 56:62 arg1 glycosylation 64:76 its protein glycosylation 52:76 its protein glycosylation in Escherichia coli 52:96 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 9 39 with transfer 1331:1338 arg1 complexity 1386:1395 greater complexity 1378:1395 greater complexity 1378:1395 This work showed that PglB protein might be able to accommodate the transfer of the further engineered glycan with greater complexity. 25449758 5 40 theme Hex 778:780 arg1 mannoses 825:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 5 40 theme Hex 778:780 arg1 residues 782:789 the Hex residues 774:789 the Hex residues 774:789 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 0 41 from glycosylation 64:76 arg1 coli 93:96 Escherichia coli 81:96 Escherichia coli 81:96 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 3 42 dep E. 516:517 arg1 coli 519:522 coli 519:522 coli 519:522 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 1 43 theme ubiquitous 120:129 arg1 modification 158:169 a ubiquitous protein post-translational modification 118:169 a ubiquitous protein post-translational modification mechanism in eukaryotes 118:193 N-Glycosylation is a ubiquitous protein post-translational modification mechanism in eukaryotes. 25449758 4 44 theme N-glycan 650:657 arg1 analysis 612:619 Mass spectrometric analysis 593:619 Mass spectrometric analysis of the fluorescently labeled N-glycan 593:657 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 25449758 8 45 theme binding 1246:1252 arg1 protein 1254:1260 maltose binding protein 1238:1260 maltose binding protein 1238:1260 Surprisingly, the Man3-GlcNAc2 structure but not Man-GlcNAc2 was transferred onto maltose binding protein. 25449758 6 46 theme early 948:952 arg1 pathway 974:980 an early eukaryotic dolichol pathway 945:980 an early eukaryotic dolichol pathway 945:980 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 7 47 theme maltose 1131:1137 arg1 protein 1147:1153 recombinant maltose binding protein 1119:1153 recombinant maltose binding protein 1119:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 1 48 theme protein 131:137 arg1 modification 158:169 a ubiquitous protein post-translational modification 118:169 a ubiquitous protein post-translational modification mechanism in eukaryotes 118:193 N-Glycosylation is a ubiquitous protein post-translational modification mechanism in eukaryotes. 25449758 4 49 theme labeled 642:648 arg1 N-glycan 650:657 the fluorescently labeled N-glycan 624:657 the fluorescently labeled N-glycan 624:657 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 25449758 7 50 theme binding 1139:1145 arg1 protein 1147:1153 recombinant maltose binding protein 1119:1153 recombinant maltose binding protein 1119:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 1 51 theme post-translational 139:156 arg1 modification 158:169 a ubiquitous protein post-translational modification 118:169 a ubiquitous protein post-translational modification mechanism in eukaryotes 118:193 N-Glycosylation is a ubiquitous protein post-translational modification mechanism in eukaryotes. 25449758 5 52 theme α1,3- 796:800 arg1 mannoses 825:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 5 52 theme α1,3- 796:800 arg1 residues 782:789 the Hex residues 774:789 the Hex residues 774:789 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 2 53 theme synthetic 212:220 arg1 pathway 222:228 a synthetic pathway 210:228 a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae 210:290 In this work, a synthetic pathway containing glycosyltransferases from Saccharomyces cerevisiae was introduced to Escherichia coli to synthesize lipid-linked mannosyl-chitobiose (Man-GlcNAc2) and trimannosyl-chitobiose (Man3-GlcNAc2). 25449758 0 54 gly glycosylation 64:76 arg1 coli 93:96 Escherichia coli 81:96 Escherichia coli 81:96 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 0 54 gly glycosylation 64:76 arg1 N-glycan 39:46 initial-stage eukaryotic N-glycan 14:46 initial-stage eukaryotic N-glycan 14:46 Production of initial-stage eukaryotic N-glycan and its protein glycosylation in Escherichia coli. 25449758 1 55 theme modification 158:169 arg1 mechanism 171:179 a ubiquitous protein post-translational modification mechanism 118:179 a ubiquitous protein post-translational modification mechanism in eukaryotes 118:193 N-Glycosylation is a ubiquitous protein post-translational modification mechanism in eukaryotes. 25449758 6 56 located observed 933:940 arg2 that 928:931 that 928:931 that 928:931 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 6 56 located observed 933:940 arg1 pathway 974:980 an early eukaryotic dolichol pathway 945:980 an early eukaryotic dolichol pathway 945:980 These results indicated that the constructed system synthesizes a Man3-GlcNAc2, identical to that observed in an early eukaryotic dolichol pathway. 25449758 7 57 theme recombinant 1119:1129 arg1 protein 1147:1153 recombinant maltose binding protein 1119:1153 recombinant maltose binding protein 1119:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 3 58 theme periplasmic 469:479 arg1 protein 481:487 a model periplasmic protein 461:487 a model periplasmic protein 461:487 Transfer of Man3-GlcNAc2 onto a model periplasmic protein occurred in the engineered E. coli cell using oligosaccharyltransferase PglB from Campylobacter jejuni. 25449758 4 59 theme spectrometric 598:610 arg1 analysis 612:619 Mass spectrometric analysis 593:619 Mass spectrometric analysis of the fluorescently labeled N-glycan 593:657 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 25449758 7 60 theme glycosylation 1096:1108 arg1 motif 1110:1114 an engineered glycosylation motif 1082:1114 an engineered glycosylation motif of recombinant maltose binding protein 1082:1153 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 5 61 link β1,4-linked 813:823 arg1 mannoses 825:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 α1,3-, α1,6- and β1,4-linked mannoses 796:832 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 5 61 link β1,4-linked 813:823 arg1 residues 782:789 the Hex residues 774:789 the Hex residues 774:789 The reversed-phase HPLC analysis suggested that the Hex residues were α1,3-, α1,6- and β1,4-linked mannoses. 25449758 7 62 theme glycopeptide 992:1003 arg1 spectrometry 1010:1021 glycopeptide mass spectrometry 992:1021 glycopeptide mass spectrometry 992:1021 Finally, glycopeptide mass spectrometry confirmed the transfer of the assembled glycan moiety onto an engineered glycosylation motif of recombinant maltose binding protein. 25449758 4 63 theme Hex 712:714 arg1 residues 716:723 2 HexNAc and 3 Hex residues 697:723 residues 716:723 Mass spectrometric analysis of the fluorescently labeled N-glycan indicated a glycan signal composed of 2 HexNAc and 3 Hex residues. 24867127 2 0 attach isolated 334:341 arg1 pigs 382:385 1-to-3 day-old neonatal wild-type pigs 348:385 1-to-3 day-old neonatal wild-type pigs 348:385 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 2 0 attach isolated 334:341 arg2 NPCCs 323:327 METHODS NPCCs 315:327 METHODS NPCCs 315:327 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 9 1 theme hybrid 1069:1074 arg1 types 1088:1092 the hybrid and complex types 1065:1092 types 1088:1092 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 1 2 theme cell 151:154 arg1 NPCCs 166:170 NPCCs 166:170 NPCCs 166:170 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 2 theme cell 151:154 arg1 clusters 156:163 neonatal porcine islet-like cell clusters 123:163 neonatal porcine islet-like cell clusters (NPCCs) 123:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 8 3 theme structures 1027:1036 arg1 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 3 theme structures 1027:1036 arg1 amounts 1003:1009 large amounts 997:1009 large amounts of high mannose structures 997:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 3 theme structures 1027:1036 arg1 feature 948:954 a feature 946:954 a feature of the N-glycans of NPCC 946:979 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 0 4 theme clusters 71:78 arg1 N-glycans 25:33 N-glycans 25:33 N-glycans of neonatal porcine islet-like cell clusters (NPCC) 25:85 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 7 5 dep RESULTS 849:855 arg1 comprised 879:887 comprised 879:887 were comprised of 14 neutral, 5 mono-sialyl and 5 di-sialyl glycans 874:940 RESULTS The NPCC glycans were comprised of 14 neutral, 5 mono-sialyl and 5 di-sialyl glycans. 24867127 2 6 theme METHODS 315:321 arg1 NPCCs 323:327 METHODS NPCCs 315:327 METHODS NPCCs 315:327 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 9 7 theme complex 1080:1086 arg1 types 1088:1092 the hybrid and complex types 1065:1092 types 1088:1092 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 3 8 theme chymotrypsin 508:519 arg1 mixture 533:539 a chymotrypsin and trypsin mixture 506:539 a chymotrypsin and trypsin mixture 506:539 The extract was proteolyzed by treatment with a chymotrypsin and trypsin mixture and further digested with glycoamidase A to release the N-glycans. 24867127 8 9 theme N-glycans 963:971 arg1 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 9 theme N-glycans 963:971 arg1 amounts 1003:1009 large amounts 997:1009 large amounts of high mannose structures 997:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 9 theme N-glycans 963:971 arg1 feature 948:954 a feature 946:954 a feature of the N-glycans of NPCC 946:979 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 4 10 theme materials 641:649 arg1 removal 618:624 the removal 614:624 the removal of the peptide materials 614:649 After the removal of the peptide materials, the reducing ends of the N-glycans were derivatized with 2-aminopyridine. 24867127 12 11 theme glycoantigen 1538:1549 arg1 studies 1523:1529 future studies 1516:1529 future studies of the glycoantigen associated with NPCC 1516:1570 CONCLUSIONS The data herein will be helpful for future studies of the glycoantigen associated with NPCC. 24867127 12 12 theme future 1516:1521 arg1 studies 1523:1529 future studies 1516:1529 future studies of the glycoantigen associated with NPCC 1516:1570 CONCLUSIONS The data herein will be helpful for future studies of the glycoantigen associated with NPCC. 24867127 2 13 theme neonatal 363:370 arg1 pigs 382:385 1-to-3 day-old neonatal wild-type pigs 348:385 1-to-3 day-old neonatal wild-type pigs 348:385 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 6 14 theme TOF-MS 832:837 arg1 analysis 839:846 MALDI TOF-MS analysis 826:846 MALDI TOF-MS analysis 826:846 PA-oligosaccharides were also subjected to MALDI TOF-MS analysis. 24867127 8 15 theme NPCC 976:979 arg1 N-glycans 963:971 the N-glycans 959:971 the N-glycans of NPCC 959:979 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 10 16 theme adult 1319:1323 arg1 islets 1325:1330 adult islets 1319:1330 adult islets 1319:1330 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 2 17 theme day-old 355:361 arg1 pigs 382:385 1-to-3 day-old neonatal wild-type pigs 348:385 1-to-3 day-old neonatal wild-type pigs 348:385 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 0 18 theme structural 2:11 arg1 analysis 13:20 A structural analysis 0:20 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 0:86 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 6 19 theme MALDI 826:830 arg1 analysis 839:846 MALDI TOF-MS analysis 826:846 MALDI TOF-MS analysis 826:846 PA-oligosaccharides were also subjected to MALDI TOF-MS analysis. 24867127 9 20 theme Fucα1-6GlcNAc 1106:1118 arg1 structure 1120:1128 a Fucα1-6GlcNAc structure 1104:1128 a Fucα1-6GlcNAc structure 1104:1128 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 4 21 theme N-glycans 677:685 arg1 ends 665:668 the reducing ends 652:668 the reducing ends of the N-glycans 652:685 After the removal of the peptide materials, the reducing ends of the N-glycans were derivatized with 2-aminopyridine. 24867127 8 22 theme mannose 1019:1025 arg1 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 3 23 theme trypsin 525:531 arg1 mixture 533:539 a chymotrypsin and trypsin mixture 506:539 a chymotrypsin and trypsin mixture 506:539 The extract was proteolyzed by treatment with a chymotrypsin and trypsin mixture and further digested with glycoamidase A to release the N-glycans. 24867127 10 24 contain contained 1209:1217 arg1 preparation 1197:1207 the NPCC preparation 1188:1207 the NPCC preparation 1188:1207 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 24 contain contained 1209:1217 arg2 glycans 1252:1258 two mono-sialyl glycans 1236:1258 two mono-sialyl glycans 1236:1258 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 24 contain contained 1209:1217 arg2 neutral 1224:1230 neutral 1224:1230 neutral 1224:1230 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 9 25 mod modified 1144:1151 arg1 types 1088:1092 the hybrid and complex types 1065:1092 types 1088:1092 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 25 mod modified 1144:1151 arg3 residues 1166:1173 sulfate residues 1158:1173 sulfate residues 1158:1173 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 25 mod modified 1144:1151 arg1 all 1058:1060 all 1058:1060 all 1058:1060 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 5 26 theme ODS 770:772 arg1 columns 774:780 ODS columns 770:780 ODS columns 770:780 This mixture was applied to DEAE, amide and ODS columns. 24867127 8 27 theme large 997:1001 arg1 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 27 theme large 997:1001 arg1 amounts 1003:1009 large amounts 997:1009 large amounts of high mannose structures 997:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 27 theme large 997:1001 arg1 feature 948:954 a feature 946:954 a feature of the N-glycans of NPCC 946:979 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 1 28 theme BACKGROUND 88:97 arg1 N-glycans 99:107 BACKGROUND N-glycans 88:107 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) 88:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 4 29 theme peptide 633:639 arg1 materials 641:649 the peptide materials 629:649 the peptide materials 629:649 After the removal of the peptide materials, the reducing ends of the N-glycans were derivatized with 2-aminopyridine. 24867127 1 30 theme adult 280:284 arg1 islets 290:295 adult pig islets 280:295 adult pig islets (APIs) 280:302 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 30 theme adult 280:284 arg1 APIs 298:301 APIs 298:301 APIs 298:301 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 8 31 theme high 1014:1017 arg1 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 12 32 dep CONCLUSIONS 1468:1478 arg1 helpful 1504:1510 helpful 1504:1510 helpful 1504:1510 CONCLUSIONS The data herein will be helpful for future studies of the glycoantigen associated with NPCC. 24867127 12 32 dep CONCLUSIONS 1468:1478 arg1 data 1484:1487 The data 1480:1487 The data 1480:1487 CONCLUSIONS The data herein will be helpful for future studies of the glycoantigen associated with NPCC. 24867127 10 33 theme NPCC 1192:1195 arg1 preparation 1197:1207 the NPCC preparation 1188:1207 the NPCC preparation 1188:1207 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 3 34 with treatment 491:499 arg1 mixture 533:539 a chymotrypsin and trypsin mixture 506:539 a chymotrypsin and trypsin mixture 506:539 The extract was proteolyzed by treatment with a chymotrypsin and trypsin mixture and further digested with glycoamidase A to release the N-glycans. 24867127 0 35 theme N-glycans 25:33 arg1 analysis 13:20 A structural analysis 0:20 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 0:86 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 1 36 theme pig 286:288 arg1 islets 290:295 adult pig islets 280:295 adult pig islets (APIs) 280:302 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 36 theme pig 286:288 arg1 APIs 298:301 APIs 298:301 APIs 298:301 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 37 from differences 226:236 arg1 antigenicity 259:270 antigenicity 259:270 antigenicity 259:270 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 37 from differences 226:236 arg1 glycosylation 241:253 glycosylation 241:253 glycosylation 241:253 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 10 38 theme mono-sialyl 1240:1250 arg1 glycans 1252:1258 two mono-sialyl glycans 1236:1258 two mono-sialyl glycans 1236:1258 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 1 39 theme mapping 192:198 arg1 technique 200:208 a mapping technique 190:208 a mapping technique 190:208 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 0 40 theme porcine 47:53 arg1 clusters 71:78 neonatal porcine islet-like cell clusters 38:78 neonatal porcine islet-like cell clusters (NPCC) 38:85 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 0 40 theme porcine 47:53 arg1 NPCC 81:84 NPCC 81:84 NPCC 81:84 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 2 41 dep Korbutt 445:451 arg1 al 456:457 Korbutt et al 445:457 Korbutt et al 445:457 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 10 42 theme human 1378:1382 arg1 islets 1384:1389 human islets 1378:1389 human islets 1378:1389 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 9 43 theme other 1046:1050 arg1 hand 1052:1055 the other hand 1042:1055 the other hand 1042:1055 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 4 44 theme reducing 656:663 arg1 ends 665:668 the reducing ends 652:668 the reducing ends of the N-glycans 652:685 After the removal of the peptide materials, the reducing ends of the N-glycans were derivatized with 2-aminopyridine. 24867127 0 45 theme neonatal 38:45 arg1 clusters 71:78 neonatal porcine islet-like cell clusters 38:78 neonatal porcine islet-like cell clusters (NPCC) 38:85 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 0 45 theme neonatal 38:45 arg1 NPCC 81:84 NPCC 81:84 NPCC 81:84 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 7 46 dep neutral 895:901 arg1 glycans 934:940 glycans 934:940 glycans 934:940 RESULTS The NPCC glycans were comprised of 14 neutral, 5 mono-sialyl and 5 di-sialyl glycans. 24867127 10 47 theme nine 1352:1355 arg1 seven 1337:1341 seven 1337:1341 seven 1337:1341 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 47 theme nine 1352:1355 arg1 nine 1352:1355 nine 1352:1355 nine 1352:1355 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 7 48 theme NPCC 861:864 arg1 glycans 866:872 The NPCC glycans 857:872 The NPCC glycans 857:872 RESULTS The NPCC glycans were comprised of 14 neutral, 5 mono-sialyl and 5 di-sialyl glycans. 24867127 9 49 theme sulfate 1158:1164 arg1 residues 1166:1173 sulfate residues 1158:1173 sulfate residues 1158:1173 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg1 types 1088:1092 the hybrid and complex types 1065:1092 types 1088:1092 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg2 structure 1120:1128 a Fucα1-6GlcNAc structure 1104:1128 a Fucα1-6GlcNAc structure 1104:1128 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg1 all 1058:1060 all 1058:1060 all 1058:1060 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg2 all 1058:1060 all 1058:1060 all 1058:1060 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg2 types 1088:1092 the hybrid and complex types 1065:1092 types 1088:1092 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 9 50 contain contained 1094:1102 arg1 hand 1052:1055 the other hand 1042:1055 the other hand 1042:1055 On the other hand, all of the hybrid and complex types contained a Fucα1-6GlcNAc structure, but were not modified with sulfate residues. 24867127 10 51 theme di-sialyl 1268:1276 arg1 glycans 1278:1284 two di-sialyl glycans 1264:1284 two di-sialyl glycans 1264:1284 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 1 52 theme neonatal 123:130 arg1 NPCCs 166:170 NPCCs 166:170 NPCCs 166:170 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 52 theme neonatal 123:130 arg1 clusters 156:163 neonatal porcine islet-like cell clusters 123:163 neonatal porcine islet-like cell clusters (NPCCs) 123:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 0 53 theme cell 66:69 arg1 clusters 71:78 neonatal porcine islet-like cell clusters 38:78 neonatal porcine islet-like cell clusters (NPCC) 38:85 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 0 53 theme cell 66:69 arg1 NPCC 81:84 NPCC 81:84 NPCC 81:84 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 2 54 theme wild-type 372:380 arg1 pigs 382:385 1-to-3 day-old neonatal wild-type pigs 348:385 1-to-3 day-old neonatal wild-type pigs 348:385 METHODS NPCCs were isolated from 1-to-3 day-old neonatal wild-type pigs and cultured for 9 days, using the technique described by Korbutt et al. 24867127 1 55 theme porcine 132:138 arg1 NPCCs 166:170 NPCCs 166:170 NPCCs 166:170 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 55 theme porcine 132:138 arg1 clusters 156:163 neonatal porcine islet-like cell clusters 123:163 neonatal porcine islet-like cell clusters (NPCCs) 123:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 8 56 contain contained 987:995 arg2 feature 948:954 a feature 946:954 a feature of the N-glycans of NPCC 946:979 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 56 contain contained 987:995 arg2 structures 1027:1036 high mannose structures 1014:1036 high mannose structures 1014:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 56 contain contained 987:995 arg1 NPCC 982:985 NPCC 982:985 NPCC 982:985 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 8 56 contain contained 987:995 arg2 amounts 1003:1009 large amounts 997:1009 large amounts of high mannose structures 997:1036 As a feature of the N-glycans of NPCC, NPCC contained large amounts of high mannose structures. 24867127 0 57 theme islet-like 55:64 arg1 clusters 71:78 neonatal porcine islet-like cell clusters 38:78 neonatal porcine islet-like cell clusters (NPCC) 38:85 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 0 57 theme islet-like 55:64 arg1 NPCC 81:84 NPCC 81:84 NPCC 81:84 A structural analysis of N-glycans of neonatal porcine islet-like cell clusters (NPCC). 24867127 1 58 attach isolated 109:116 arg2 N-glycans 99:107 BACKGROUND N-glycans 88:107 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) 88:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 58 attach isolated 109:116 arg1 NPCCs 166:170 NPCCs 166:170 NPCCs 166:170 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 58 attach isolated 109:116 arg1 clusters 156:163 neonatal porcine islet-like cell clusters 123:163 neonatal porcine islet-like cell clusters (NPCCs) 123:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 3 59 theme glycoamidase 567:578 arg1 A 580:580 glycoamidase A 567:580 glycoamidase A 567:580 The extract was proteolyzed by treatment with a chymotrypsin and trypsin mixture and further digested with glycoamidase A to release the N-glycans. 24867127 10 60 located detected 1366:1373 arg2 nine 1352:1355 nine 1352:1355 nine 1352:1355 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 60 located detected 1366:1373 arg2 seven 1337:1341 seven 1337:1341 seven 1337:1341 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 60 located detected 1366:1373 arg1 islets 1384:1389 human islets 1378:1389 human islets 1378:1389 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 1 61 theme islet-like 140:149 arg1 NPCCs 166:170 NPCCs 166:170 NPCCs 166:170 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 1 61 theme islet-like 140:149 arg1 clusters 156:163 neonatal porcine islet-like cell clusters 123:163 neonatal porcine islet-like cell clusters (NPCCs) 123:171 BACKGROUND N-glycans isolated from neonatal porcine islet-like cell clusters (NPCCs) were analyzed by a mapping technique, to examine the differences in glycosylation and antigenicity between adult pig islets (APIs) and NPCCs. 24867127 10 62 located found 1310:1314 arg2 neutral 1224:1230 neutral 1224:1230 neutral 1224:1230 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 62 located found 1310:1314 arg1 islets 1325:1330 adult islets 1319:1330 adult islets 1319:1330 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24867127 10 62 located found 1310:1314 arg2 glycans 1252:1258 two mono-sialyl glycans 1236:1258 two mono-sialyl glycans 1236:1258 Among them, the NPCC preparation contained five neutral and two mono-sialyl glycans and two di-sialyl glycans that were not typically found in adult islets, and seven of these nine were not detected in human islets. 24812685 0 0 theme G 112:112 arg1 N-glycans 114:122 immunoglobulin G N-glycans 97:122 immunoglobulin G N-glycans of ovarian cancer patients 97:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 5 1 theme available 791:799 arg1 A2G2S2 843:848 A2G2S2 843:848 A2G2S2 843:848 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 5 1 theme available 791:799 arg1 oligosaccharide 826:840 the commercially available disialylated biantennary oligosaccharide 774:840 the commercially available disialylated biantennary oligosaccharide (A2G2S2) 774:849 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 8 2 theme patients 1137:1144 arg1 profiles 1125:1132 the IgG N-glycan profiles 1108:1132 the IgG N-glycan profiles of patients suffering from ovarian cancer 1108:1174 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 0 3 theme immunoglobulin 97:110 arg1 N-glycans 114:122 immunoglobulin G N-glycans 97:122 immunoglobulin G N-glycans of ovarian cancer patients 97:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 7 4 theme FA2B[3 996:1001 arg1 G1S1 1003:1006 FA2B[3]G1S1 996:1006 FA2B[3]G1S1 996:1006 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 3 5 theme negative 560:567 arg1 charge 569:574 their negative charge 554:574 their negative charge 554:574 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 9 6 with impossible 1474:1483 arg1 method 1507:1512 the conventional method 1490:1512 the conventional method 1490:1512 Finally, using the methylation protocol, we could further demonstrate an increase of A2, which was technically impossible with the conventional method. 24812685 8 7 dep conventional 1186:1197 arg1 the 1182:1184 the 1182:1184 the 1182:1184 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 8 8 from decrease 1322:1329 arg1 structures 1351:1360 digalactosylated structures 1334:1360 digalactosylated structures 1334:1360 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 0 9 theme N-glycans 114:122 arg1 analyses 85:92 capillary electrophoresis laser-induced fluorescence analyses 32:92 capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients 32:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 1 10 theme diagnostic 291:300 arg1 Alterations 152:162 Alterations 152:162 Alterations in IgG N-glycosylation 152:185 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 1 10 theme diagnostic 291:300 arg1 markers 302:308 diagnostic markers 291:308 diagnostic markers 291:308 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 0 11 theme cancer 135:140 arg1 patients 142:149 ovarian cancer patients 127:149 ovarian cancer patients 127:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 3 12 theme due 547:549 arg1 times 541:545 short migration times 525:545 short migration times due to their negative charge 525:574 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 3 13 theme short 525:529 arg1 times 541:545 short migration times 525:545 short migration times due to their negative charge 525:574 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 8 14 theme agalactosylated 1269:1283 arg1 structures 1285:1294 agalactosylated structures 1269:1294 agalactosylated structures 1269:1294 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 0 15 theme ovarian 127:133 arg1 patients 142:149 ovarian cancer patients 127:149 ovarian cancer patients 127:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 5 16 theme biantennary 814:824 arg1 A2G2S2 843:848 A2G2S2 843:848 A2G2S2 843:848 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 5 16 theme biantennary 814:824 arg1 oligosaccharide 826:840 the commercially available disialylated biantennary oligosaccharide 774:840 the commercially available disialylated biantennary oligosaccharide (A2G2S2) 774:849 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 1 17 from Alterations 152:162 arg1 N-glycosylation 171:185 IgG N-glycosylation 167:185 IgG N-glycosylation 167:185 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 8 18 theme methylation 1203:1213 arg1 methods.With 1215:1226 methylation methods.With 1203:1226 methylation methods.With 1203:1226 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 0 19 theme acid 7:10 arg1 methylation 12:22 Sialic acid methylation 0:22 Sialic acid methylation 0:22 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 7 20 theme FA2G2S1 983:989 arg1 co-migration 963:974 co-migration 963:974 co-migration of A2, FA2G2S1, and FA2B[3]G1S1 963:1006 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 2 21 theme N-glycans 366:374 arg1 CE-LIF 311:316 CE-LIF 311:316 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 311:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 21 theme N-glycans 366:374 arg1 method 404:409 a well-established rapid method 379:409 a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material 379:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 7 22 theme N-glycans 1072:1080 arg1 quantification 1048:1061 an accurate quantification 1036:1061 an accurate quantification of these N-glycans 1036:1080 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 0 23 theme Sialic 0:5 arg1 acid 7:10 Sialic acid 0:10 Sialic acid methylation 0:22 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 2 24 theme labeled 358:364 arg1 N-glycans 366:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 8 25 theme structures 1285:1294 arg1 increase 1257:1264 an increase 1254:1264 an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures 1254:1360 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 9 26 theme methylation 1382:1392 arg1 protocol 1394:1401 the methylation protocol 1378:1401 the methylation protocol 1378:1401 Finally, using the methylation protocol, we could further demonstrate an increase of A2, which was technically impossible with the conventional method. 24812685 5 27 theme acids 720:724 arg1 charge 703:708 the negative charge 690:708 the negative charge of sialic acids 690:724 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 7 28 theme A2 979:980 arg1 co-migration 963:974 co-migration 963:974 co-migration of A2, FA2G2S1, and FA2B[3]G1S1 963:1006 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 2 29 theme acid 353:356 arg1 N-glycans 366:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 1 30 theme number 222:227 arg1 development 205:215 the development 201:215 the development of a number of diseases including cancer 201:256 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 2 31 theme material 481:488 arg1 amounts 461:467 only low amounts 452:467 only low amounts of starting material 452:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 31 theme material 481:488 arg1 material 481:488 starting material 472:488 starting material 472:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 0 32 theme patients 142:149 arg1 N-glycans 114:122 immunoglobulin G N-glycans 97:122 immunoglobulin G N-glycans of ovarian cancer patients 97:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 2 33 theme 8-aminopyrene-1,3,6-trisulfonic 321:351 arg1 N-glycans 366:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 321:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 1 34 theme IgG 167:169 arg1 N-glycosylation 171:185 IgG N-glycosylation 167:185 IgG N-glycosylation 167:185 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 0 35 theme electrophoresis 42:56 arg1 analyses 85:92 capillary electrophoresis laser-induced fluorescence analyses 32:92 capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients 32:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 8 36 theme N-glycan 1116:1123 arg1 profiles 1125:1132 the IgG N-glycan profiles 1108:1132 the IgG N-glycan profiles of patients suffering from ovarian cancer 1108:1174 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 3 37 theme sialylated 499:508 arg1 N-glycans 510:518 sialylated N-glycans 499:518 sialylated N-glycans 499:518 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 9 38 theme conventional 1494:1505 arg1 method 1507:1512 the conventional method 1490:1512 the conventional method 1490:1512 Finally, using the methylation protocol, we could further demonstrate an increase of A2, which was technically impossible with the conventional method. 24812685 1 39 theme diseases 232:239 arg1 cancer 251:256 cancer 251:256 cancer 251:256 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 1 39 theme diseases 232:239 arg1 number 222:227 a number 220:227 a number of diseases including cancer 220:256 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 0 40 theme capillary 32:40 arg1 analyses 85:92 capillary electrophoresis laser-induced fluorescence analyses 32:92 capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients 32:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 8 41 theme IgG 1112:1114 arg1 profiles 1125:1132 the IgG N-glycan profiles 1108:1132 the IgG N-glycan profiles of patients suffering from ovarian cancer 1108:1174 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 1 42 used used 283:286 arg2 markers 302:308 diagnostic markers 291:308 diagnostic markers 291:308 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 1 42 used used 283:286 arg2 Alterations 152:162 Alterations 152:162 Alterations in IgG N-glycosylation 152:185 Alterations in IgG N-glycosylation coincide with the development of a number of diseases including cancer and could potentially be used as diagnostic markers. 24812685 2 43 theme low 457:459 arg1 amounts 461:467 only low amounts 452:467 only low amounts of starting material 452:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 43 theme low 457:459 arg1 material 481:488 starting material 472:488 starting material 472:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 0 44 theme fluorescence 72:83 arg1 analyses 85:92 capillary electrophoresis laser-induced fluorescence analyses 32:92 capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients 32:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 5 45 theme negative 694:701 arg1 charge 703:708 the negative charge 690:708 the negative charge of sialic acids 690:724 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 3 46 contain have 520:523 arg1 N-glycans 510:518 sialylated N-glycans 499:518 sialylated N-glycans 499:518 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 3 46 contain have 520:523 arg2 times 541:545 short migration times 525:545 short migration times due to their negative charge 525:574 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 7 47 theme accurate 1039:1046 arg1 quantification 1048:1061 an accurate quantification 1036:1061 an accurate quantification of these N-glycans 1036:1080 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 2 48 theme rapid 398:402 arg1 CE-LIF 311:316 CE-LIF 311:316 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 311:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 48 theme rapid 398:402 arg1 method 404:409 a well-established rapid method 379:409 a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material 379:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 0 49 theme laser-induced 58:70 arg1 analyses 85:92 capillary electrophoresis laser-induced fluorescence analyses 32:92 capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients 32:149 Sialic acid methylation refines capillary electrophoresis laser-induced fluorescence analyses of immunoglobulin G N-glycans of ovarian cancer patients. 24812685 5 50 dep available 791:799 arg1 disialylated 801:812 disialylated 801:812 disialylated 801:812 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 3 51 gly sialylated 499:508 arg1 N-glycans 510:518 sialylated N-glycans 499:518 sialylated N-glycans 499:518 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 2 52 theme well-established 381:396 arg1 CE-LIF 311:316 CE-LIF 311:316 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans 311:374 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 52 theme well-established 381:396 arg1 method 404:409 a well-established rapid method 379:409 a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material 379:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 6 53 attach isolated 865:872 arg1 serum 893:897 healthy human serum 879:897 healthy human serum 879:897 IgGN-glycans isolated from healthy human serum were then analyzed using this method. 24812685 6 53 attach isolated 865:872 arg2 IgGN-glycans 852:863 IgGN-glycans 852:863 IgGN-glycans isolated from healthy human serum 852:897 IgGN-glycans isolated from healthy human serum were then analyzed using this method. 24812685 6 54 theme human 887:891 arg1 serum 893:897 healthy human serum 879:897 healthy human serum 879:897 IgGN-glycans isolated from healthy human serum were then analyzed using this method. 24812685 3 55 theme migration 531:539 arg1 times 541:545 short migration times 525:545 short migration times due to their negative charge 525:574 However,sialylated N-glycans have short migration times due to their negative charge. 24812685 5 56 theme sialic 713:718 arg1 acids 720:724 sialic acids 713:724 sialic acids 713:724 In this work, we neutralized the negative charge of sialic acids by methylation and optimized the protocol using the commercially available disialylated biantennary oligosaccharide (A2G2S2). 24812685 6 57 theme healthy 879:885 arg1 serum 893:897 healthy human serum 879:897 healthy human serum 879:897 IgGN-glycans isolated from healthy human serum were then analyzed using this method. 24812685 8 58 theme digalactosylated 1334:1349 arg1 structures 1351:1360 digalactosylated structures 1334:1360 digalactosylated structures 1334:1360 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 8 59 theme ovarian 1161:1167 arg1 cancer 1169:1174 ovarian cancer 1161:1174 ovarian cancer 1161:1174 Finally, we investigated the IgG N-glycan profiles of patients suffering from ovarian cancer using the conventional and methylation methods.With both methods, we observed an increase of agalactosylated structures that was accompanied by a decrease in digalactosylated structures. 24812685 2 60 theme starting 472:479 arg1 material 481:488 starting material 472:488 starting material 472:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 2 61 theme IgG 427:429 arg1 N-glycans 431:439 IgG N-glycans 427:439 IgG N-glycans that needs only low amounts of starting material 427:488 CE-LIF of 8-aminopyrene-1,3,6-trisulfonic acid labeled N-glycans is a well-established rapid method to characterize IgG N-glycans that needs only low amounts of starting material. 24812685 9 62 theme A2 1448:1449 arg1 impossible 1474:1483 impossible 1474:1483 impossible 1474:1483 Finally, using the methylation protocol, we could further demonstrate an increase of A2, which was technically impossible with the conventional method. 24812685 9 62 theme A2 1448:1449 arg1 increase 1436:1443 an increase 1433:1443 an increase 1433:1443 Finally, using the methylation protocol, we could further demonstrate an increase of A2, which was technically impossible with the conventional method. 24812685 7 63 theme G1S1 1003:1006 arg1 co-migration 963:974 co-migration 963:974 co-migration of A2, FA2G2S1, and FA2B[3]G1S1 963:1006 We could demonstrate that co-migration of A2, FA2G2S1, and FA2B[3]G1S1 was prevented,which allowed an accurate quantification of these N-glycans. 24812685 4 64 theme neutral 644:650 arg1 glycans 652:658 neutral glycans 644:658 neutral glycans 644:658 As a result,some of them are not well resolved and co-migrate with neutral glycans. 26003531 4 0 theme emerging 542:549 arg1 trends 551:556 the emerging trends 538:556 the emerging trends in glycomics research 538:578 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 5 1 theme aminoxyTMT 705:714 arg1 reagents 716:723 aminoxyTMT reagents 705:723 aminoxyTMT reagents 705:723 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 4 2 theme based 636:640 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 6 3 theme ionization 1003:1012 arg1 ESI-MS 1033:1038 ESI-MS 1033:1038 ESI-MS 1033:1038 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 3 theme ionization 1003:1012 arg1 spectrometry 1019:1030 electrospray ionization mass spectrometry 990:1030 electrospray ionization mass spectrometry (ESI-MS) 990:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 4 4 theme glycomics 561:569 arg1 research 571:578 glycomics research 561:578 glycomics research 561:578 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 8 5 theme differential 1317:1328 arg1 analysis 1330:1337 differential analysis 1317:1337 differential analysis across biological samples 1317:1363 This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. 26003531 7 6 theme generated 1208:1216 arg1 data 1218:1221 generated data 1208:1221 generated data 1208:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 4 7 theme spectrometry 623:634 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 5 8 theme glycan 798:803 arg1 efficiency 816:825 glycan ionization efficiency 798:825 glycan ionization efficiency 798:825 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 9 9 theme such 1415:1418 arg1 protocols 1420:1428 such protocols 1415:1428 such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures 1415:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 4 10 from trends 551:556 arg1 research 571:578 glycomics research 561:578 glycomics research 561:578 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 0 11 with reagents 88:95 arg1 SimGlycan 102:110 SimGlycan 102:110 SimGlycan 102:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 3 12 theme growing 422:428 arg1 importance 430:439 the growing importance 418:439 the growing importance of glycoproteomics 418:458 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 5 13 theme ionization 805:814 arg1 efficiency 816:825 glycan ionization efficiency 798:825 glycan ionization efficiency 798:825 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 6 14 theme -coupled 978:985 arg1 LC 975:976 LC 975:976 LC 975:976 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 14 theme -coupled 978:985 arg1 chromatography 959:972 liquid chromatography 952:972 liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS) 952:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 7 15 theme quantitative 1183:1194 arg1 analysis 1196:1203 the qualitative and quantitative analysis 1163:1203 the qualitative and quantitative analysis of generated data 1163:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 2 16 theme glycans 306:312 arg1 structure 293:301 structure 293:301 structure 293:301 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 2 16 theme glycans 306:312 arg1 abundance 279:287 abundance 279:287 abundance 279:287 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 2 17 from Monitoring 250:259 arg1 structure 293:301 structure 293:301 structure 293:301 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 2 17 from Monitoring 250:259 arg1 abundance 279:287 abundance 279:287 abundance 279:287 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 4 18 theme quantitative 642:653 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 1 19 theme critical 193:200 arg1 roles 202:206 critical roles 193:206 critical roles 193:206 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 7 20 theme data 1218:1221 arg1 analysis 1196:1203 the qualitative and quantitative analysis 1163:1203 the qualitative and quantitative analysis of generated data 1163:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 9 21 theme quantitative 1434:1445 arg1 analysis 1447:1454 quantitative analysis 1434:1454 quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures 1434:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 3 22 theme in-depth 473:480 arg1 studies 511:517 in-depth qualitative and quantitative studies 473:517 in-depth qualitative and quantitative studies of glycans 473:528 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 7 23 theme tools 1145:1149 arg1 one 1073:1075 one 1073:1075 one 1073:1075 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 7 23 theme tools 1145:1149 arg1 lack 1125:1128 the lack 1121:1128 the lack of informatics tools to automate the qualitative and quantitative analysis of generated data 1121:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 7 23 theme tools 1145:1149 arg1 challenges 1090:1099 the major challenges 1080:1099 the major challenges left unaddressed 1080:1116 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 2 24 from changes 264:270 arg1 structure 293:301 structure 293:301 structure 293:301 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 2 24 from changes 264:270 arg1 abundance 279:287 abundance 279:287 abundance 279:287 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 6 25 theme mass 1014:1017 arg1 ESI-MS 1033:1038 ESI-MS 1033:1038 ESI-MS 1033:1038 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 25 theme mass 1014:1017 arg1 spectrometry 1019:1030 electrospray ionization mass spectrometry 990:1030 electrospray ionization mass spectrometry (ESI-MS) 990:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 8 26 theme glycans 1288:1294 arg1 analysis 1330:1337 differential analysis 1317:1337 differential analysis across biological samples 1317:1363 This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. 26003531 8 26 theme glycans 1288:1294 arg1 identification/quantitation 1257:1283 identification/quantitation 1257:1283 identification/quantitation of glycans using MS/MS data 1257:1311 This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. 26003531 10 27 from Proteomics 1567:1576 arg1 India 1581:1585 India 1581:1585 India 1581:1585 This article is part of a Special Issue entitled: Proteomics in India. 26003531 10 28 theme Special 1543:1549 arg1 Issue 1551:1555 a Special Issue 1541:1555 a Special Issue entitled 1541:1564 This article is part of a Special Issue entitled: Proteomics in India. 26003531 7 29 from lack 1125:1128 arg1 proteomics 1061:1070 proteomics 1061:1070 proteomics 1061:1070 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 6 30 theme electrospray 990:1001 arg1 ESI-MS 1033:1038 ESI-MS 1033:1038 ESI-MS 1033:1038 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 30 theme electrospray 990:1001 arg1 spectrometry 1019:1030 electrospray ionization mass spectrometry 990:1030 electrospray ionization mass spectrometry (ESI-MS) 990:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 31 theme quantitative 895:906 arg1 analysis 908:915 quantitative analysis 895:915 quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS) 895:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 8 32 theme biological 1346:1355 arg1 samples 1357:1363 biological samples 1346:1363 biological samples 1346:1363 This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. 26003531 2 33 dep abundance 279:287 arg1 the 275:277 the 275:277 the 275:277 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 0 34 theme quantitative 35:46 arg1 glycomics 48:56 mass spectrometry-based quantitative glycomics 11:56 mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan 11:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 4 35 theme related 598:604 arg1 trends 551:556 the emerging trends 538:556 the emerging trends in glycomics research 538:578 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 4 35 theme related 598:604 arg1 innovation 587:596 the innovation 583:596 the innovation related to accurate mass spectrometry based quantitative analysis of glycans 583:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 4 35 theme related 598:604 arg1 One 531:533 One 531:533 One 531:533 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 7 36 theme informatics 1133:1143 arg1 tools 1145:1149 informatics tools 1133:1149 informatics tools 1133:1149 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 1 37 theme Protein 113:119 arg1 modification 166:177 a common post-translational modification 138:177 a common post-translational modification 138:177 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 1 37 theme Protein 113:119 arg1 glycosylation 121:133 Protein glycosylation 113:133 Protein glycosylation 113:133 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 1 38 theme biological 215:224 arg1 processes 226:234 the biological processes 211:234 the biological processes of organisms 211:247 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 0 39 theme spectrometry-based 16:33 arg1 glycomics 48:56 mass spectrometry-based quantitative glycomics 11:56 mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan 11:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 4 40 theme mass 618:621 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 5 41 theme carbohydrates 774:786 arg1 quantitation 758:769 efficient relative quantitation 739:769 efficient relative quantitation of carbohydrates 739:786 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 9 42 theme labeled-glycans 1470:1484 arg1 analysis 1447:1454 quantitative analysis 1434:1454 quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures 1434:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 3 43 theme glycans 522:528 arg1 studies 511:517 in-depth qualitative and quantitative studies 473:517 in-depth qualitative and quantitative studies of glycans 473:528 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 4 44 theme accurate 609:616 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 6 45 theme liquid 952:957 arg1 LC 975:976 LC 975:976 LC 975:976 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 6 45 theme liquid 952:957 arg1 chromatography 959:972 liquid chromatography 952:972 liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS) 952:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 2 46 theme changes 264:270 arg1 Monitoring 250:259 Monitoring 250:259 Monitoring of changes in the abundance and structure of glycans 250:312 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 2 47 from abundance 279:287 arg1 Monitoring 250:259 Monitoring 250:259 Monitoring of changes in the abundance and structure of glycans 250:312 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 3 48 theme quantitative 498:509 arg1 studies 511:517 in-depth qualitative and quantitative studies 473:517 in-depth qualitative and quantitative studies of glycans 473:528 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 3 49 theme glycoproteomics 444:458 arg1 importance 430:439 the growing importance 418:439 the growing importance of glycoproteomics 418:458 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 8 50 theme MS/MS 1302:1306 arg1 data 1308:1311 MS/MS data 1302:1311 MS/MS data 1302:1311 This analysis typically includes identification/quantitation of glycans using MS/MS data and differential analysis across biological samples. 26003531 7 51 theme major 1084:1088 arg1 challenges 1090:1099 the major challenges 1080:1099 the major challenges left unaddressed 1080:1116 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 1 52 theme organisms 239:247 arg1 processes 226:234 the biological processes 211:234 the biological processes of organisms 211:247 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 0 53 theme tandem 72:77 arg1 reagents 88:95 aminoxy tandem mass tag reagents 64:95 aminoxy tandem mass tag reagents with SimGlycan 64:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 9 54 theme complex 1499:1505 arg1 mixtures 1507:1514 complex mixtures 1499:1514 complex mixtures 1499:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 3 55 theme qualitative 482:492 arg1 studies 511:517 in-depth qualitative and quantitative studies 473:517 in-depth qualitative and quantitative studies of glycans 473:528 Hence, the growing importance of glycoproteomics necessitates in-depth qualitative and quantitative studies of glycans. 26003531 7 56 theme challenges 1090:1099 arg1 one 1073:1075 one 1073:1075 one 1073:1075 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 7 56 theme challenges 1090:1099 arg1 lack 1125:1128 the lack 1121:1128 the lack of informatics tools to automate the qualitative and quantitative analysis of generated data 1121:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 7 56 theme challenges 1090:1099 arg1 challenges 1090:1099 the major challenges 1080:1099 the major challenges left unaddressed 1080:1116 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 2 57 theme protein 367:373 arg1 glycosylation 375:387 protein glycosylation 367:387 protein glycosylation 367:387 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 0 58 theme aminoxy 64:70 arg1 reagents 88:95 aminoxy tandem mass tag reagents 64:95 aminoxy tandem mass tag reagents with SimGlycan 64:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 5 59 theme efficient 739:747 arg1 quantitation 758:769 efficient relative quantitation 739:769 efficient relative quantitation of carbohydrates 739:786 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 6 60 theme direct 933:938 arg1 infusion 940:947 direct infusion 933:947 direct infusion 933:947 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 7 61 theme qualitative 1167:1177 arg1 analysis 1196:1203 the qualitative and quantitative analysis 1163:1203 the qualitative and quantitative analysis of generated data 1163:1221 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 9 62 attach derived 1486:1492 arg1 mixtures 1507:1514 complex mixtures 1499:1514 complex mixtures 1499:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 9 62 attach derived 1486:1492 arg2 labeled-glycans 1470:1484 aminoxyTMT labeled-glycans 1459:1484 aminoxyTMT labeled-glycans derived from complex mixtures 1459:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 5 63 theme relative 749:756 arg1 quantitation 758:769 efficient relative quantitation 739:769 efficient relative quantitation of carbohydrates 739:786 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 10 64 theme Issue 1551:1555 arg1 part 1533:1536 part 1533:1536 part of a Special Issue entitled 1533:1564 This article is part of a Special Issue entitled: Proteomics in India. 26003531 5 65 theme analytical 841:850 arg1 throughput 852:861 analytical throughput 841:861 analytical throughput 841:861 Recently, we have introduced aminoxyTMT reagents, which enable efficient relative quantitation of carbohydrates, improved glycan ionization efficiency and increased analytical throughput. 26003531 1 66 theme common 140:145 arg1 modification 166:177 a common post-translational modification 138:177 a common post-translational modification 138:177 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 1 66 theme common 140:145 arg1 glycosylation 121:133 Protein glycosylation 113:133 Protein glycosylation 113:133 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 2 67 from structure 293:301 arg1 Monitoring 250:259 Monitoring 250:259 Monitoring of changes in the abundance and structure of glycans 250:312 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 26003531 4 68 theme glycans 667:673 arg1 analysis 655:662 accurate mass spectrometry based quantitative analysis 609:662 accurate mass spectrometry based quantitative analysis of glycans 609:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 7 69 theme unaddressed 1106:1116 arg1 challenges 1090:1099 the major challenges 1080:1099 the major challenges left unaddressed 1080:1116 However, unlike in proteomics, one of the major challenges left unaddressed is the lack of informatics tools to automate the qualitative and quantitative analysis of generated data. 26003531 6 70 theme N-glycans 920:928 arg1 analysis 908:915 quantitative analysis 895:915 quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS) 895:1039 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 9 71 theme aminoxyTMT 1459:1468 arg1 labeled-glycans 1470:1484 aminoxyTMT labeled-glycans 1459:1484 aminoxyTMT labeled-glycans derived from complex mixtures 1459:1514 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 1 72 theme post-translational 147:164 arg1 modification 166:177 a common post-translational modification 138:177 a common post-translational modification 138:177 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 1 72 theme post-translational 147:164 arg1 glycosylation 121:133 Protein glycosylation 113:133 Protein glycosylation 113:133 Protein glycosylation is a common post-translational modification, which serves critical roles in the biological processes of organisms. 26003531 0 73 theme tag 84:86 arg1 reagents 88:95 aminoxy tandem mass tag reagents 64:95 aminoxy tandem mass tag reagents with SimGlycan 64:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 9 74 theme software 1384:1391 arg1 modules 1393:1399 software modules 1384:1399 software modules 1384:1399 We have developed software modules to streamline such protocols for quantitative analysis of aminoxyTMT labeled-glycans derived from complex mixtures. 26003531 6 75 used used 886:889 arg2 reagents 870:877 These reagents 864:877 These reagents 864:877 These reagents can be used for quantitative analysis of N-glycans by direct infusion or liquid chromatography (LC)-coupled to electrospray ionization mass spectrometry (ESI-MS). 26003531 0 76 theme mass 79:82 arg1 reagents 88:95 aminoxy tandem mass tag reagents 64:95 aminoxy tandem mass tag reagents with SimGlycan 64:110 Automating mass spectrometry-based quantitative glycomics using aminoxy tandem mass tag reagents with SimGlycan. 26003531 4 77 theme trends 551:556 arg1 trends 551:556 the emerging trends 538:556 the emerging trends in glycomics research 538:578 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 4 77 theme trends 551:556 arg1 innovation 587:596 the innovation 583:596 the innovation related to accurate mass spectrometry based quantitative analysis of glycans 583:673 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 4 77 theme trends 551:556 arg1 One 531:533 One 531:533 One 531:533 One of the emerging trends in glycomics research is the innovation related to accurate mass spectrometry based quantitative analysis of glycans. 26003531 2 78 theme various 393:399 arg1 diseases 401:408 various diseases 393:408 various diseases 393:408 Monitoring of changes in the abundance and structure of glycans may be necessary to explain the correlations between protein glycosylation and various diseases. 29042829 4 0 theme amino 957:961 arg1 sequences 968:976 amino acid sequences 957:976 amino acid sequences 957:976 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 3 1 theme accurate 663:670 arg1 quantitation 672:683 accurate quantitation 663:683 accurate quantitation 663:683 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 8 2 theme release 1476:1482 arg1 kinetics 1484:1491 release kinetics 1476:1491 release kinetics 1476:1491 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 4 3 from differences 942:952 arg1 sequences 968:976 amino acid sequences 957:976 amino acid sequences 957:976 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 2 4 attach attached 557:564 arg1 residue 598:604 the reducing terminal GlcNAc residue 569:604 the reducing terminal GlcNAc residue 569:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 4 attach attached 557:564 arg2 fucose 550:555 a 3-linked fucose 539:555 a 3-linked fucose attached to the reducing terminal GlcNAc residue 539:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 4 5 theme slight 935:940 arg1 differences 942:952 the slight differences 931:952 the slight differences in amino acid sequences 931:976 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 3 6 theme glycan 644:649 arg1 release 651:657 complete glycan release 635:657 complete glycan release 635:657 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 1 7 theme biologic 115:122 arg1 activity 124:131 The biologic activity 111:131 The biologic activity of IgG molecules 111:148 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 3 8 theme acid 834:837 arg1 sequence 839:846 amino acid sequence 828:846 amino acid sequence 828:846 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 0 9 from Kinetics 0:7 arg1 IgG 58:60 IgG 58:60 IgG 58:60 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 0 9 from Kinetics 0:7 arg1 G 55:55 Human Immunoglobulin G 34:55 Human Immunoglobulin G (IgG) 34:61 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 2 10 theme 3-linked 541:548 arg1 fucose 550:555 a 3-linked fucose 539:555 a 3-linked fucose attached to the reducing terminal GlcNAc residue 539:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 5 11 gly nonfucosylated 1205:1218 arg1 etc 1264:1266 etc 1264:1266 etc. 1264:1267 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 3 12 theme sequence 839:846 arg1 effect 797:802 the effect 793:802 the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs 793:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 0 13 theme PNGase 66:71 arg1 F 73:73 PNGase F 66:73 PNGase F 66:73 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 0 14 from G 55:55 arg1 Kinetics 0:7 Kinetics 0:7 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F 0:73 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 0 14 from G 55:55 arg1 Release 21:27 N-Glycan Release 12:27 N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F 12:73 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 5 15 dep etc 1264:1266 arg1 i.e. 1199:1202 i.e. 1199:1202 i.e. 1199:1202 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 3 16 theme glycan 863:868 arg1 release 870:876 glycan release 863:876 glycan release from glycopeptides of IgGs 863:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 2 17 theme innermost 459:467 arg1 GlcNAc 490:495 GlcNAc 490:495 GlcNAc 490:495 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 17 theme innermost 459:467 arg1 N-acetylglucosamine 469:487 innermost N-acetylglucosamine 459:487 innermost N-acetylglucosamine (GlcNAc) 459:496 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 5 18 theme significant 1065:1075 arg1 differences 1077:1087 statistically significant differences 1051:1087 statistically significant differences in the deglycosylation rate constants 1051:1125 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 2 19 theme terminal 582:589 arg1 residue 598:604 the reducing terminal GlcNAc residue 569:604 the reducing terminal GlcNAc residue 569:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 20 theme GlcNAc 591:596 arg1 residue 598:604 the reducing terminal GlcNAc residue 569:604 the reducing terminal GlcNAc residue 569:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 7 21 theme Similar 1362:1368 arg1 reductions 1370:1379 Similar reductions 1362:1379 Similar reductions in rate 1362:1387 Similar reductions in rate were associated with the presence of a bisecting-GlcNAc. 29042829 3 22 theme release 870:876 arg1 rate 855:858 the rate 851:858 the rate of glycan release from glycopeptides of IgGs 851:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 4 23 theme acid 963:966 arg1 sequences 968:976 amino acid sequences 957:976 amino acid sequences 957:976 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 2 24 theme reducing 573:580 arg1 residue 598:604 the reducing terminal GlcNAc residue 569:604 the reducing terminal GlcNAc residue 569:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 8 25 theme glycosylation 1548:1560 arg1 study 1562:1566 the glycosylation study 1544:1566 the glycosylation study of IgGs 1544:1574 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 5 26 theme nonfucosylated 1205:1218 arg1 etc 1264:1266 etc 1264:1266 etc. 1264:1267 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 5 27 from differences 1077:1087 arg1 constants 1117:1125 the deglycosylation rate constants 1092:1125 the deglycosylation rate constants 1092:1125 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 5 28 theme glycan 1181:1186 arg1 structure 1188:1196 glycan structure 1181:1196 glycan structure 1181:1196 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 2 29 dep approach 279:286 arg1 analyze 291:297 analyze 291:297 to analyze glycosylation of IgGs 288:319 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 1 30 theme IgG 136:138 arg1 molecules 140:148 IgG molecules 136:148 IgG molecules 136:148 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 3 31 from effect 797:802 arg1 rate 855:858 the rate 851:858 the rate of glycan release from glycopeptides of IgGs 851:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 7 32 theme bisecting-GlcNAc 1428:1443 arg1 presence 1414:1421 the presence 1410:1421 the presence of a bisecting-GlcNAc 1410:1443 Similar reductions in rate were associated with the presence of a bisecting-GlcNAc. 29042829 8 33 theme IgGs 1571:1574 arg1 study 1562:1566 the glycosylation study 1544:1566 the glycosylation study of IgGs 1544:1574 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 1 34 theme molecules 140:148 arg1 activity 124:131 The biologic activity 111:131 The biologic activity of IgG molecules 111:148 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 0 35 theme Release 21:27 arg1 Kinetics 0:7 Kinetics 0:7 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F 0:73 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 3 36 theme IgGs 900:903 arg1 release 870:876 glycan release 863:876 glycan release from glycopeptides of IgGs 863:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 2 37 theme enzyme 366:371 arg1 F 395:395 the enzyme peptide N-glycosidase F 362:395 the enzyme peptide N-glycosidase F 362:395 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 38 theme peptide 373:379 arg1 F 395:395 the enzyme peptide N-glycosidase F 362:395 the enzyme peptide N-glycosidase F 362:395 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 1 39 theme IgG 237:239 arg1 glycosylation 241:253 IgG glycosylation 237:253 IgG glycosylation 237:253 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 6 40 theme acid 1300:1303 arg1 residue 1305:1311 a single sialic acid residue 1284:1311 a single sialic acid residue 1284:1311 For example, a single sialic acid residue was found to decrease the rate by a factor of 3. 29042829 8 41 theme significant 1505:1515 arg1 variations 1530:1539 significant quantitative variations 1505:1539 significant quantitative variations of the glycosylation study of IgGs 1505:1574 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 0 42 theme Human 34:38 arg1 IgG 58:60 IgG 58:60 IgG 58:60 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 0 42 theme Human 34:38 arg1 G 55:55 Human Immunoglobulin G 34:55 Human Immunoglobulin G (IgG) 34:61 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 1 43 theme glycosylation 241:253 arg1 critical 258:265 critical 258:265 critical 258:265 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 1 43 theme glycosylation 241:253 arg1 analysis 225:232 the analysis 221:232 the analysis of IgG glycosylation 221:253 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 2 44 gly glycosylation 299:311 arg1 IgGs 316:319 IgGs 316:319 IgGs 316:319 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 4 45 theme deglycosylation 1020:1034 arg1 rate 1036:1039 a statistically different deglycosylation rate 994:1039 a statistically different deglycosylation rate 994:1039 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 2 46 contain containing 528:537 arg1 those 522:526 those 522:526 those 522:526 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 46 contain containing 528:537 arg2 fucose 550:555 a 3-linked fucose 539:555 a 3-linked fucose attached to the reducing terminal GlcNAc residue 539:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 6 47 theme 3 1359:1359 arg1 factor 1349:1354 a factor 1347:1354 a factor of 3 1347:1359 For example, a single sialic acid residue was found to decrease the rate by a factor of 3. 29042829 3 48 theme reaction 745:752 arg1 kinetics 711:718 the kinetics 707:718 the kinetics of this de-glycosylation reaction for IgG glycopeptides 707:774 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 6 49 theme single 1286:1291 arg1 residue 1305:1311 a single sialic acid residue 1284:1311 a single sialic acid residue 1284:1311 For example, a single sialic acid residue was found to decrease the rate by a factor of 3. 29042829 5 50 dep nonfucosylated 1205:1218 arg1 sialylated 1252:1261 sialylated 1252:1261 sialylated 1252:1261 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 5 50 dep nonfucosylated 1205:1218 arg1 bisecting-GlcNAc 1234:1249 bisecting-GlcNAc 1234:1249 bisecting-GlcNAc 1234:1249 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 5 50 dep nonfucosylated 1205:1218 arg1 fucosylated 1221:1231 fucosylated 1221:1231 fucosylated 1221:1231 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 2 51 theme N-glycosidase 381:393 arg1 F 395:395 the enzyme peptide N-glycosidase F 362:395 the enzyme peptide N-glycosidase F 362:395 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 52 theme N-glycans 349:357 arg1 release 334:340 the release 330:340 the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue 330:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 5 53 theme deglycosylation 1096:1110 arg1 constants 1117:1125 the deglycosylation rate constants 1092:1125 the deglycosylation rate constants 1092:1125 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 6 54 theme sialic 1293:1298 arg1 residue 1305:1311 a single sialic acid residue 1284:1311 a single sialic acid residue 1284:1311 For example, a single sialic acid residue was found to decrease the rate by a factor of 3. 29042829 5 55 theme rate 1112:1115 arg1 constants 1117:1125 the deglycosylation rate constants 1092:1125 the deglycosylation rate constants 1092:1125 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 0 56 theme Immunoglobulin 40:53 arg1 IgG 58:60 IgG 58:60 IgG 58:60 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 0 56 theme Immunoglobulin 40:53 arg1 G 55:55 Human Immunoglobulin G 34:55 Human Immunoglobulin G (IgG) 34:61 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 3 57 gly glycopeptides 883:895 arg2 glycopeptides 883:895 glycopeptides 883:895 glycopeptides 883:895 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 8 58 theme study 1562:1566 arg1 variations 1530:1539 significant quantitative variations 1505:1539 significant quantitative variations of the glycosylation study of IgGs 1505:1574 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 2 59 link 3-linked 541:548 arg1 fucose 550:555 a 3-linked fucose 539:555 a 3-linked fucose attached to the reducing terminal GlcNAc residue 539:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 3 60 theme de-glycosylation 728:743 arg1 reaction 745:752 this de-glycosylation reaction 723:752 this de-glycosylation reaction 723:752 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 61 theme complete 635:642 arg1 release 651:657 complete glycan release 635:657 complete glycan release 635:657 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 62 from rate 855:858 arg1 glycopeptides 883:895 glycopeptides 883:895 glycopeptides 883:895 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 63 from glycopeptides 883:895 arg1 rate 855:858 the rate 851:858 the rate of glycan release from glycopeptides of IgGs 851:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 63 from glycopeptides 883:895 arg1 release 870:876 glycan release 863:876 glycan release from glycopeptides of IgGs 863:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 64 theme amino 828:832 arg1 sequence 839:846 amino acid sequence 828:846 amino acid sequence 828:846 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 1 65 theme crystallizable 170:183 arg1 N-glycosylation 194:208 its crystallizable fragment N-glycosylation 166:208 its crystallizable fragment N-glycosylation 166:208 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 2 66 theme N-glycans 505:513 arg1 GlcNAc 490:495 GlcNAc 490:495 GlcNAc 490:495 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 66 theme N-glycans 505:513 arg1 residue 447:453 the asparagine residue 432:453 the asparagine residue 432:453 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 66 theme N-glycans 505:513 arg1 N-acetylglucosamine 469:487 innermost N-acetylglucosamine 459:487 innermost N-acetylglucosamine (GlcNAc) 459:496 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 66 theme N-glycans 505:513 arg1 N-glycans 505:513 all N-glycans 501:513 all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue 501:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 3 67 gly glycopeptides 762:774 arg2 glycopeptides 762:774 IgG glycopeptides 758:774 IgG glycopeptides 758:774 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 2 68 theme standard 270:277 arg1 approach 279:286 A standard approach 268:286 A standard approach to analyze glycosylation of IgGs 268:319 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 3 69 theme glycan 807:812 arg1 structure 814:822 glycan structure 807:822 glycan structure 807:822 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 1 70 theme fragment 185:192 arg1 N-glycosylation 194:208 its crystallizable fragment N-glycosylation 166:208 its crystallizable fragment N-glycosylation 166:208 The biologic activity of IgG molecules is modulated by its crystallizable fragment N-glycosylation, and thus, the analysis of IgG glycosylation is critical. 29042829 5 71 dep differing 1163:1171 arg1 etc 1264:1266 etc 1264:1266 etc. 1264:1267 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 4 72 theme different 1010:1018 arg1 rate 1036:1039 a statistically different deglycosylation rate 994:1039 a statistically different deglycosylation rate 994:1039 This study revealed that the slight differences in amino acid sequences did not lead to a statistically different deglycosylation rate. 29042829 7 73 from reductions 1370:1379 arg1 rate 1384:1387 rate 1384:1387 rate 1384:1387 Similar reductions in rate were associated with the presence of a bisecting-GlcNAc. 29042829 8 74 from differences 1461:1471 arg1 kinetics 1484:1491 release kinetics 1476:1491 release kinetics 1476:1491 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 29042829 3 75 theme structure 814:822 arg1 effect 797:802 the effect 793:802 the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs 793:903 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 3 76 theme IgG 758:760 arg1 glycopeptides 762:774 IgG glycopeptides 758:774 IgG glycopeptides 758:774 The importance of obtaining complete glycan release for accurate quantitation led us to investigate the kinetics of this de-glycosylation reaction for IgG glycopeptides and to determine the effect of glycan structure and amino acid sequence on the rate of glycan release from glycopeptides of IgGs. 29042829 2 77 theme asparagine 436:445 arg1 residue 447:453 the asparagine residue 432:453 the asparagine residue 432:453 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 2 77 theme asparagine 436:445 arg1 N-glycans 505:513 all N-glycans 501:513 all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue 501:604 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 0 78 theme N-Glycan 12:19 arg1 Release 21:27 N-Glycan Release 12:27 N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F 12:73 Kinetics of N-Glycan Release from Human Immunoglobulin G (IgG) by PNGase F: All Glycans Are Not Created Equal. 29042829 5 79 gly glycopeptides 1149:1161 arg2 glycopeptides 1149:1161 glycopeptides 1149:1161 glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.) 1149:1268 However, statistically significant differences in the deglycosylation rate constants were observed between glycopeptides differing only in glycan structure (i.e., nonfucosylated, fucosylated, bisecting-GlcNAc, sialylated, etc.). 29042829 2 80 theme IgGs 316:319 arg1 glycosylation 299:311 glycosylation 299:311 glycosylation of IgGs 299:319 A standard approach to analyze glycosylation of IgGs involves the release of the N-glycans by the enzyme peptide N-glycosidase F, which cleaves the linkage between the asparagine residue and innermost N-acetylglucosamine (GlcNAc) of all N-glycans except those containing a 3-linked fucose attached to the reducing terminal GlcNAc residue. 29042829 8 81 theme quantitative 1517:1528 arg1 variations 1530:1539 significant quantitative variations 1505:1539 significant quantitative variations of the glycosylation study of IgGs 1505:1574 We predict the differences in release kinetics can lead to significant quantitative variations of the glycosylation study of IgGs. 25040827 1 0 theme N-glycan 154:161 arg1 structure 137:145 The predominant structure 121:145 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 121:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 0 theme N-glycan 154:161 arg1 GlcNAc-OH 309:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 1 from N-glycan 154:161 arg1 hormone 189:195 the prothoracicotropic hormone 166:195 the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 166:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 1 from N-glycan 154:161 arg1 PTTH 198:201 PTTH 198:201 PTTH 198:201 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 2 theme silkmoths 245:253 arg1 heads 236:240 1.8 million adult heads 218:240 1.8 million adult heads of silkmoths 218:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 0 3 theme silkmoth 99:106 arg1 mori 115:118 the silkmoth Bombyx mori 95:118 the silkmoth Bombyx mori 95:118 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 0 4 theme Bombyx 108:113 arg1 mori 115:118 the silkmoth Bombyx mori 95:118 the silkmoth Bombyx mori 95:118 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 1 5 theme prothoracicotropic 170:187 arg1 hormone 189:195 the prothoracicotropic hormone 166:195 the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 166:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 5 theme prothoracicotropic 170:187 arg1 PTTH 198:201 PTTH 198:201 PTTH 198:201 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 6 theme baculovirus-expressed 354:374 arg1 PTTH 388:391 the baculovirus-expressed recombinant PTTH 350:391 the baculovirus-expressed recombinant PTTH 350:391 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 7 from structure 137:145 arg1 hormone 189:195 the prothoracicotropic hormone 166:195 the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 166:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 7 from structure 137:145 arg1 PTTH 198:201 PTTH 198:201 PTTH 198:201 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 0 8 theme Structural 0:9 arg1 determination 11:23 Structural determination 0:23 Structural determination of an N-glycan moiety 0:45 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 0 9 attach attached 47:54 arg1 hormone 82:88 the prothoracicotropic hormone 59:88 the prothoracicotropic hormone from the silkmoth Bombyx mori 59:118 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 0 9 attach attached 47:54 arg2 determination 11:23 Structural determination 0:23 Structural determination of an N-glycan moiety 0:45 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 2 10 theme N-glycosylated 441:454 arg1 PTTH 456:459 N-glycosylated PTTH 441:459 N-glycosylated PTTH 441:459 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 2 11 theme expression 580:589 arg1 system 591:596 an Escherichia coli expression system 560:596 an Escherichia coli expression system 560:596 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 1 12 theme Manα1-6Manβ1-4GlcNAcβ1-4 276:299 arg1 structure 137:145 The predominant structure 121:145 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 121:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 12 theme Manα1-6Manβ1-4GlcNAcβ1-4 276:299 arg1 GlcNAc-OH 309:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 13 theme recombinant 376:386 arg1 PTTH 388:391 the baculovirus-expressed recombinant PTTH 350:391 the baculovirus-expressed recombinant PTTH 350:391 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 0 14 theme moiety 40:45 arg1 determination 11:23 Structural determination 0:23 Structural determination of an N-glycan moiety 0:45 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 2 15 theme Escherichia 563:573 arg1 system 591:596 an Escherichia coli expression system 560:596 an Escherichia coli expression system 560:596 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 2 16 theme progression 405:415 arg1 assay 417:421 An ecdysis progression assay 394:421 An ecdysis progression assay 394:421 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 0 17 theme N-glycan 31:38 arg1 moiety 40:45 an N-glycan moiety 28:45 an N-glycan moiety 28:45 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 1 18 theme Fucα1-6 301:307 arg1 structure 137:145 The predominant structure 121:145 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 121:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 18 theme Fucα1-6 301:307 arg1 GlcNAc-OH 309:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 2 19 theme higher 482:487 arg1 activity 489:496 a slightly higher activity 471:496 a slightly higher activity than the recombinant PTTH 471:522 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 2 20 theme ecdysis 397:403 arg1 assay 417:421 An ecdysis progression assay 394:421 An ecdysis progression assay 394:421 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 2 21 dep Escherichia 563:573 arg1 coli 575:578 coli 575:578 coli 575:578 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 0 22 from mori 115:118 arg1 hormone 82:88 the prothoracicotropic hormone 59:88 the prothoracicotropic hormone from the silkmoth Bombyx mori 59:118 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 1 23 theme predominant 125:135 arg1 structure 137:145 The predominant structure 121:145 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 121:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 23 theme predominant 125:135 arg1 GlcNAc-OH 309:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 24 from hormone 189:195 arg1 structure 137:145 The predominant structure 121:145 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 121:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 24 from hormone 189:195 arg1 GlcNAc-OH 309:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH 276:317 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 25 theme million 222:228 arg1 heads 236:240 1.8 million adult heads 218:240 1.8 million adult heads of silkmoths 218:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 0 26 theme prothoracicotropic 63:80 arg1 hormone 82:88 the prothoracicotropic hormone 59:88 the prothoracicotropic hormone from the silkmoth Bombyx mori 59:118 Structural determination of an N-glycan moiety attached to the prothoracicotropic hormone from the silkmoth Bombyx mori. 25040827 1 27 attach isolated 204:211 arg1 heads 236:240 1.8 million adult heads 218:240 1.8 million adult heads of silkmoths 218:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 27 attach isolated 204:211 arg2 PTTH 198:201 PTTH 198:201 PTTH 198:201 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 27 attach isolated 204:211 arg2 hormone 189:195 the prothoracicotropic hormone 166:195 the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths 166:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 1 28 theme adult 230:234 arg1 heads 236:240 1.8 million adult heads 218:240 1.8 million adult heads of silkmoths 218:253 The predominant structure of the N-glycan on the prothoracicotropic hormone (PTTH) isolated from 1.8 million adult heads of silkmoths was determined to be Manα1-6Manβ1-4GlcNAcβ1-4(Fucα1-6)GlcNAc-OH, which is identical to that of the baculovirus-expressed recombinant PTTH. 25040827 2 29 theme recombinant 507:517 arg1 PTTH 519:522 the recombinant PTTH 503:522 the recombinant PTTH 503:522 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 25040827 2 30 gly N-glycosylated 441:454 arg1 PTTH 456:459 N-glycosylated PTTH 441:459 N-glycosylated PTTH 441:459 An ecdysis progression assay demonstrated that N-glycosylated PTTH exhibited a slightly higher activity than the recombinant PTTH without N-glycosylation produced by an Escherichia coli expression system. 28585749 4 0 theme cervical 869:876 arg1 cancer 878:883 uterine cervical cancer 861:883 uterine cervical cancer 861:883 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 7 1 from tissues 1199:1205 arg1 lectin 1158:1163 the best lectin 1149:1163 the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis 1149:1251 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 1 from tissues 1199:1205 arg1 It 1142:1143 It 1142:1143 It 1142:1143 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 9 2 theme microarray 1588:1597 arg1 technique 1599:1607 The present lectin microarray technique 1569:1607 The present lectin microarray technique 1569:1607 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 3 3 theme lectin 515:520 arg1 microarray 522:531 A lectin microarray 513:531 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans 505:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 7 4 theme discriminating 1169:1182 arg1 SCC 1184:1186 discriminating SCC 1169:1186 discriminating SCC 1169:1186 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 9 5 theme tissue-based 1630:1641 arg1 analysis 1652:1659 tissue-based glycomic analysis 1630:1659 tissue-based glycomic analysis of various tumors 1630:1677 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 1 6 theme proteins 189:196 arg1 Glycosylation 172:184 AIM Glycosylation 168:184 AIM Glycosylation of proteins 168:196 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 3 7 theme squamous 733:740 arg1 NSE 754:756 NSE 754:756 NSE 754:756 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 3 7 theme squamous 733:740 arg1 epithelium 742:751 normal squamous epithelium 726:751 normal squamous epithelium (NSE) 726:757 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 0 8 theme squamous 121:128 arg1 carcinoma 135:143 squamous cell carcinoma 121:143 squamous cell carcinoma of the uterine cervix 121:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 9 9 theme various 1664:1670 arg1 tumors 1672:1677 various tumors 1664:1677 various tumors 1664:1677 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 9 10 dep CONCLUSION 1558:1567 arg1 applied 1618:1624 applied 1618:1624 could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers 1609:1724 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 9 11 theme present 1573:1579 arg1 technique 1599:1607 The present lectin microarray technique 1569:1607 The present lectin microarray technique 1569:1607 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 7 12 theme best 1153:1156 arg1 lectin 1158:1163 the best lectin 1149:1163 the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis 1149:1251 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 12 theme best 1153:1156 arg1 It 1142:1143 It 1142:1143 It 1142:1143 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 1 13 theme cancer 212:217 arg1 cells 219:223 cancer cells 212:223 cancer cells 212:223 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 8 14 theme specific 1391:1398 arg1 staining 1412:1419 specific cytoplasmic staining 1391:1419 specific cytoplasmic staining of SCC cells 1391:1432 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 3 15 theme different 560:568 arg1 preferences 578:588 different binding preferences 560:588 different binding preferences that covered N- and O-linked glycans 560:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 3 16 dep METHODS 505:511 arg1 microarray 522:531 A lectin microarray 513:531 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans 505:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 0 17 theme cervix 160:165 arg1 carcinoma 135:143 squamous cell carcinoma 121:143 squamous cell carcinoma of the uterine cervix 121:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 6 18 theme Hippeastrum 1034:1044 arg1 lectin 1053:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 6 18 theme Hippeastrum 1034:1044 arg1 marker 1088:1093 a sensitive marker 1076:1093 a sensitive marker for distinguishing SCC of the cervix from NSE 1076:1139 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 9 19 theme glycan-related 1700:1713 arg1 biomarkers 1715:1724 glycan-related biomarkers 1700:1724 glycan-related biomarkers 1700:1724 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 8 20 theme SCC 1424:1426 arg1 cells 1428:1432 SCC cells 1424:1432 SCC cells 1424:1432 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 7 21 theme curve 1238:1242 arg1 analysis 1244:1251 receiver-operator curve analysis 1220:1251 receiver-operator curve analysis 1220:1251 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 1 22 theme glycan 336:341 arg1 profile 343:349 the complete glycan profile 323:349 the complete glycan profile of particular tumors 323:370 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 8 23 theme hybrid 1449:1454 arg1 lectin 1456:1461 Hippeastrum hybrid lectin 1437:1461 Hippeastrum hybrid lectin 1437:1461 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 1 24 theme tumors 365:370 arg1 profile 343:349 the complete glycan profile 323:349 the complete glycan profile of particular tumors 323:370 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 6 25 theme sensitive 1078:1086 arg1 lectin 1053:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 6 25 theme sensitive 1078:1086 arg1 marker 1088:1093 a sensitive marker 1076:1093 a sensitive marker for distinguishing SCC of the cervix from NSE 1076:1139 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 0 26 theme hybrid 78:83 arg1 lectin 85:90 Hippeastrum hybrid lectin 66:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin 0:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 0 27 theme Glycan 0:5 arg1 biomarker 107:115 a sensitive biomarker 95:115 a sensitive biomarker for squamous cell carcinoma of the uterine cervix 95:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 0 27 theme Glycan 0:5 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin 0:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 1 28 theme specific 279:286 arg1 cancers 288:294 specific cancers 279:294 specific cancers 279:294 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 2 29 theme carcinoma 423:431 arg1 analysis 397:404 glycomic analysis 388:404 glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix 388:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 8 30 theme little 1480:1485 arg1 staining 1487:1494 little staining 1480:1494 little staining of cervical intraepithelial neoplasia 1480:1532 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 2 31 theme squamous 409:416 arg1 SCC 434:436 SCC 434:436 SCC 434:436 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 2 31 theme squamous 409:416 arg1 carcinoma 423:431 squamous cell carcinoma 409:431 squamous cell carcinoma (SCC) of the uterine cervix 409:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 3 32 with lectins 547:553 arg1 preferences 578:588 different binding preferences 560:588 different binding preferences that covered N- and O-linked glycans 560:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 2 33 theme uterine 446:452 arg1 cervix 454:459 the uterine cervix 442:459 the uterine cervix 442:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 8 34 theme intraepithelial 1508:1522 arg1 neoplasia 1524:1532 cervical intraepithelial neoplasia 1499:1532 cervical intraepithelial neoplasia 1499:1532 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 0 35 theme Hippeastrum 66:76 arg1 lectin 85:90 Hippeastrum hybrid lectin 66:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin 0:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 3 36 theme fluorescent 671:681 arg1 detection 683:691 evanescent field-activated fluorescent detection 644:691 evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix 644:771 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 4 37 theme Formalin-fixed 774:787 arg1 specimens 815:823 Formalin-fixed, paraffin-embedded tissue specimens 774:823 Formalin-fixed, paraffin-embedded tissue specimens 774:823 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 3 38 theme evanescent 644:653 arg1 detection 683:691 evanescent field-activated fluorescent detection 644:691 evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix 644:771 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 1 39 theme distinctive 229:239 arg1 structures 248:257 distinctive glycan structures 229:257 distinctive glycan structures 229:257 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 9 40 theme lectin 1581:1586 arg1 technique 1599:1607 The present lectin microarray technique 1569:1607 The present lectin microarray technique 1569:1607 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 3 41 link O-linked 610:617 arg1 glycans 619:625 O-linked glycans 610:625 O-linked glycans 610:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 8 42 theme NSE 1553:1555 arg1 staining 1541:1548 no staining 1538:1548 no staining of NSE 1538:1555 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 8 42 theme NSE 1553:1555 arg1 staining 1487:1494 little staining 1480:1494 little staining of cervical intraepithelial neoplasia 1480:1532 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 1 43 gly Glycosylation 172:184 arg1 cells 219:223 cancer cells 212:223 cancer cells 212:223 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 1 43 gly Glycosylation 172:184 arg1 proteins 189:196 proteins 189:196 proteins 189:196 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 4 44 theme uterine 861:867 arg1 cancer 878:883 uterine cervical cancer 861:883 uterine cervical cancer 861:883 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 3 45 theme normal 726:731 arg1 NSE 754:756 NSE 754:756 NSE 754:756 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 3 45 theme normal 726:731 arg1 epithelium 742:751 normal squamous epithelium 726:751 normal squamous epithelium (NSE) 726:757 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 1 46 theme AIM 168:170 arg1 Glycosylation 172:184 AIM Glycosylation 168:184 AIM Glycosylation of proteins 168:196 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 3 47 theme SCC 718:720 arg1 analysis 706:713 glycomic analysis 697:713 glycomic analysis of SCC and normal squamous epithelium (NSE) 697:757 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 4 48 dep Formalin-fixed 774:787 arg1 paraffin-embedded 790:806 paraffin-embedded 790:806 paraffin-embedded 790:806 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 9 49 theme glycomic 1643:1650 arg1 analysis 1652:1659 tissue-based glycomic analysis 1630:1659 tissue-based glycomic analysis of various tumors 1630:1677 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 0 50 theme cell 130:133 arg1 carcinoma 135:143 squamous cell carcinoma 121:143 squamous cell carcinoma of the uterine cervix 121:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 3 51 theme epithelium 742:751 arg1 analysis 706:713 glycomic analysis 697:713 glycomic analysis of SCC and normal squamous epithelium (NSE) 697:757 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 3 52 theme binding 570:576 arg1 preferences 578:588 different binding preferences 560:588 different binding preferences that covered N- and O-linked glycans 560:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 9 53 theme tumors 1672:1677 arg1 analysis 1652:1659 tissue-based glycomic analysis 1630:1659 tissue-based glycomic analysis of various tumors 1630:1677 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 8 54 theme cytoplasmic 1400:1410 arg1 staining 1412:1419 specific cytoplasmic staining 1391:1419 specific cytoplasmic staining of SCC cells 1391:1432 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 7 55 theme large 1329:1333 arg1 0.8182 1357:1362 0.8182 1357:1362 0.8182 1357:1362 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 55 theme large 1329:1333 arg1 area 1335:1338 a large area 1327:1338 a large area under the curve (0.8182) 1327:1363 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 56 theme other 1193:1197 arg1 tissues 1199:1205 other tissues 1193:1205 other tissues according to receiver-operator curve analysis 1193:1251 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 0 57 theme uterine 152:158 arg1 cervix 160:165 the uterine cervix 148:165 the uterine cervix 148:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 5 58 theme non-tumor 924:932 arg1 tissues 934:940 non-tumor tissues 924:940 non-tumor tissues 924:940 Sections that included both tumor and non-tumor tissues were examined to detect alterations of glycans based on the lectin-binding pattern. 28585749 6 59 theme hybrid 1046:1051 arg1 lectin 1053:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 6 59 theme hybrid 1046:1051 arg1 marker 1088:1093 a sensitive marker 1076:1093 a sensitive marker for distinguishing SCC of the cervix from NSE 1076:1139 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 8 60 theme cells 1428:1432 arg1 staining 1412:1419 specific cytoplasmic staining 1391:1419 specific cytoplasmic staining of SCC cells 1391:1432 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 7 61 theme receiver-operator 1220:1236 arg1 analysis 1244:1251 receiver-operator curve analysis 1220:1251 receiver-operator curve analysis 1220:1251 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 6 62 theme RESULTS 1026:1032 arg1 lectin 1053:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin 1026:1058 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 6 62 theme RESULTS 1026:1032 arg1 marker 1088:1093 a sensitive marker 1076:1093 a sensitive marker for distinguishing SCC of the cervix from NSE 1076:1139 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 9 63 theme biomarkers 1715:1724 arg1 discovery 1687:1695 discovery 1687:1695 discovery of glycan-related biomarkers 1687:1724 CONCLUSION The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers. 28585749 8 64 theme Hippeastrum 1437:1447 arg1 lectin 1456:1461 Hippeastrum hybrid lectin 1437:1461 Hippeastrum hybrid lectin 1437:1461 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 1 65 theme complete 327:334 arg1 profile 343:349 the complete glycan profile 323:349 the complete glycan profile of particular tumors 323:370 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 4 66 with patients 847:854 arg1 cancer 878:883 uterine cervical cancer 861:883 uterine cervical cancer 861:883 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 2 67 theme cervix 454:459 arg1 SCC 434:436 SCC 434:436 SCC 434:436 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 2 67 theme cervix 454:459 arg1 carcinoma 423:431 squamous cell carcinoma 409:431 squamous cell carcinoma (SCC) of the uterine cervix 409:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 0 68 theme sensitive 97:105 arg1 biomarker 107:115 a sensitive biomarker 95:115 a sensitive biomarker for squamous cell carcinoma of the uterine cervix 95:165 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 0 68 theme sensitive 97:105 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin 0:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 2 69 theme useful 489:494 arg1 markers 496:502 useful markers 489:502 useful markers 489:502 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 1 70 theme particular 354:363 arg1 tumors 365:370 particular tumors 354:370 particular tumors 354:370 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 5 71 theme glycans 981:987 arg1 alterations 966:976 alterations 966:976 alterations of glycans based on the lectin-binding pattern 966:1023 Sections that included both tumor and non-tumor tissues were examined to detect alterations of glycans based on the lectin-binding pattern. 28585749 7 72 theme high 1297:1300 arg1 % 1319:1319 70.1% 1315:1319 70.1% 1315:1319 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 72 theme high 1297:1300 arg1 specificity 1302:1312 a high specificity 1295:1312 a high specificity (70.1%) 1295:1320 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 3 73 contain containing 533:542 arg2 lectins 547:553 45 lectins 544:553 45 lectins with different binding preferences that covered N- and O-linked glycans 544:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 3 73 contain containing 533:542 arg1 microarray 522:531 A lectin microarray 513:531 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans 505:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 0 74 theme formalin-fixed 23:36 arg1 tissues 57:63 formalin-fixed, paraffin-embedded tissues 23:63 formalin-fixed, paraffin-embedded tissues 23:63 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 0 75 dep profiling 7:15 arg1 lectin 85:90 Hippeastrum hybrid lectin 66:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin 0:90 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 2 76 theme cell 418:421 arg1 SCC 434:436 SCC 434:436 SCC 434:436 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 2 76 theme cell 418:421 arg1 carcinoma 423:431 squamous cell carcinoma 409:431 squamous cell carcinoma (SCC) of the uterine cervix 409:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 8 77 theme cervical 1499:1506 arg1 neoplasia 1524:1532 cervical intraepithelial neoplasia 1499:1532 cervical intraepithelial neoplasia 1499:1532 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 8 78 theme neoplasia 1524:1532 arg1 staining 1541:1548 no staining 1538:1548 no staining of NSE 1538:1555 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 8 78 theme neoplasia 1524:1532 arg1 staining 1487:1494 little staining 1480:1494 little staining of cervical intraepithelial neoplasia 1480:1532 Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. 28585749 3 79 theme O-linked 610:617 arg1 glycans 619:625 O-linked glycans 610:625 O-linked glycans 610:625 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 4 80 theme tissue 808:813 arg1 specimens 815:823 Formalin-fixed, paraffin-embedded tissue specimens 774:823 Formalin-fixed, paraffin-embedded tissue specimens 774:823 Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. 28585749 2 81 theme glycomic 388:395 arg1 analysis 397:404 glycomic analysis 388:404 glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix 388:459 In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. 28585749 7 82 theme high 1269:1272 arg1 % 1291:1291 81.8% 1287:1291 81.8% 1287:1291 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 7 82 theme high 1269:1272 arg1 sensitivity 1274:1284 a high sensitivity 1267:1284 a high sensitivity (81.8%) 1267:1292 It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). 28585749 3 83 theme field-activated 655:669 arg1 detection 683:691 evanescent field-activated fluorescent detection 644:691 evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix 644:771 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 0 84 dep formalin-fixed 23:36 arg1 paraffin-embedded 39:55 paraffin-embedded 39:55 paraffin-embedded 39:55 Glycan profiling using formalin-fixed, paraffin-embedded tissues: Hippeastrum hybrid lectin is a sensitive biomarker for squamous cell carcinoma of the uterine cervix. 28585749 6 85 theme cervix 1125:1130 arg1 SCC 1114:1116 SCC 1114:1116 SCC of the cervix 1114:1130 RESULTS Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. 28585749 5 86 theme lectin-binding 1002:1015 arg1 pattern 1017:1023 the lectin-binding pattern 998:1023 the lectin-binding pattern 998:1023 Sections that included both tumor and non-tumor tissues were examined to detect alterations of glycans based on the lectin-binding pattern. 28585749 3 87 theme glycomic 697:704 arg1 analysis 706:713 glycomic analysis 697:713 glycomic analysis of SCC and normal squamous epithelium (NSE) 697:757 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 28585749 1 88 theme glycan 241:246 arg1 structures 248:257 distinctive glycan structures 229:257 distinctive glycan structures 229:257 AIM Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. 28585749 3 89 theme cervix 766:771 arg1 detection 683:691 evanescent field-activated fluorescent detection 644:691 evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix 644:771 METHODS A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. 24417605 6 0 located found 840:844 arg2 type 774:777 a biantennary complex type glycan 752:784 a biantennary complex type glycan 752:784 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 0 located found 840:844 arg1 glycoproteins 858:870 multiple glycoproteins 849:870 multiple glycoproteins using site-specific glycosylation analysis 849:913 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 0 located found 840:844 arg2 N-glycan 739:746 The most abundant N-glycan 721:746 The most abundant N-glycan 721:746 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 1 1 theme lipoprotein 141:151 arg1 particles 159:167 high density lipoprotein (HDL) particles 128:167 high density lipoprotein (HDL) particles 128:167 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 9 2 theme lipid 1355:1359 arg1 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 2 theme lipid 1355:1359 arg1 tails 1361:1365 heterogeneous ceramide lipid tails 1332:1365 heterogeneous ceramide lipid tails 1332:1365 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 2 theme lipid 1355:1359 arg1 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 7 3 theme A. 1100:1101 arg1 GM3 1103:1105 fetuin A. GM3 1093:1105 fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) 1093:1151 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 3 theme A. 1100:1101 arg1 gangliosides 1228:1239 the major gangliosides 1218:1239 the major gangliosides in HDL 1218:1246 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 3 theme A. 1100:1101 arg1 CIII 1073:1076 apolipoprotein CIII 1058:1076 apolipoprotein CIII (ApoC-III) 1058:1087 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 2 4 theme sequential 305:314 arg1 micro-ultracentrifugation 316:340 sequential micro-ultracentrifugation 305:340 sequential micro-ultracentrifugation 305:340 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 10 5 theme sialylated 1518:1527 arg1 particles 1529:1537 highly sialylated particles 1511:1537 highly sialylated particles 1511:1537 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 10 5 theme sialylated 1518:1527 arg1 HDL 1503:1505 HDL 1503:1505 HDL 1503:1505 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 6 6 theme glycan 779:784 arg1 N-glycan 739:746 The most abundant N-glycan 721:746 The most abundant N-glycan 721:746 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 6 theme glycan 779:784 arg1 type 774:777 a biantennary complex type glycan 752:784 a biantennary complex type glycan 752:784 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 7 theme complex 766:772 arg1 N-glycan 739:746 The most abundant N-glycan 721:746 The most abundant N-glycan 721:746 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 7 theme complex 766:772 arg1 type 774:777 a biantennary complex type glycan 752:784 a biantennary complex type glycan 752:784 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 3 8 theme mass 531:534 arg1 spectrometry 536:547 flight mass spectrometry 524:547 flight mass spectrometry 524:547 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 3 9 theme MS/MS 553:557 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 5 10 theme neuraminic 694:703 arg1 acids 705:709 one or two neuraminic acids 683:709 one or two neuraminic acids (Neu5Ac) 683:718 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 5 10 theme neuraminic 694:703 arg1 Neu5Ac 712:717 Neu5Ac 712:717 Neu5Ac 712:717 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 3 11 theme spectrometry 536:547 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 4 12 gly sialylated 598:607 arg1 particles 564:572 HDL particles 560:572 HDL particles 560:572 HDL particles were found to be highly sialylated. 24417605 0 13 gly sialylated 61:70 arg1 particle 72:79 a highly sialylated particle 52:79 a highly sialylated particle 52:79 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 8 14 theme % 1253:1253 arg1 GM3 1255:1257 60% GM3 1251:1257 60% GM3 1251:1257 A 60% GM3 and 40% GD3 distribution was observed. 24417605 1 15 gly glycosylated 185:196 arg1 lipids 118:123 lipids 118:123 lipids 118:123 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 15 gly glycosylated 185:196 arg1 proteins 105:112 functional proteins 94:112 functional proteins 94:112 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 15 gly glycosylated 185:196 arg1 Many 82:85 Many 82:85 Many 82:85 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 7 16 from gangliosides 1228:1239 arg1 HDL 1244:1246 HDL 1244:1246 HDL 1244:1246 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 1 17 dep proteins 105:112 arg1 the 90:92 the 90:92 the 90:92 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 2 18 theme glycolipid 372:381 arg1 analysis 383:390 glycolipid analysis 372:390 glycolipid analysis 372:390 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 7 19 theme fetuin 1093:1098 arg1 GM3 1103:1105 fetuin A. GM3 1093:1105 fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) 1093:1151 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 19 theme fetuin 1093:1098 arg1 gangliosides 1228:1239 the major gangliosides 1218:1239 the major gangliosides in HDL 1218:1246 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 19 theme fetuin 1093:1098 arg1 CIII 1073:1076 apolipoprotein CIII 1058:1076 apolipoprotein CIII (ApoC-III) 1058:1087 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 2 20 attach isolated 281:288 arg1 plasma 295:300 plasma 295:300 plasma 295:300 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 2 20 attach isolated 281:288 arg2 HDL 273:275 HDL 273:275 HDL 273:275 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 7 21 theme apolipoprotein 1058:1071 arg1 GM3 1103:1105 fetuin A. GM3 1093:1105 fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) 1093:1151 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 21 theme apolipoprotein 1058:1071 arg1 gangliosides 1228:1239 the major gangliosides 1218:1239 the major gangliosides in HDL 1218:1246 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 21 theme apolipoprotein 1058:1071 arg1 ApoC-III 1079:1086 ApoC-III 1079:1086 ApoC-III 1079:1086 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 21 theme apolipoprotein 1058:1071 arg1 CIII 1073:1076 apolipoprotein CIII 1058:1076 apolipoprotein CIII (ApoC-III) 1058:1087 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 1 22 from lipids 118:123 arg1 particles 159:167 high density lipoprotein (HDL) particles 128:167 high density lipoprotein (HDL) particles 128:167 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 3 23 theme flight 524:529 arg1 spectrometry 536:547 flight mass spectrometry 524:547 flight mass spectrometry 524:547 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 7 24 theme observed 920:927 arg1 O-glycans 929:937 The observed O-glycans 916:937 The observed O-glycans 916:937 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 3 25 with analysis 476:483 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 0 26 theme Glycomic 0:7 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of high density lipoprotein 0:44 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 1 27 theme HDL 253:255 arg1 glycoconjugates 234:248 the glycoconjugates 230:248 the glycoconjugates of HDL 230:255 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 2 28 gly glycoprotein 355:366 arg1 glycoprotein 355:366 glycoprotein 355:366 glycoprotein 355:366 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 1 29 from proteins 105:112 arg1 particles 159:167 high density lipoprotein (HDL) particles 128:167 high density lipoprotein (HDL) particles 128:167 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 2 30 dep isolated 281:288 arg1 followed 343:350 followed 343:350 followed by glycoprotein and glycolipid analysis 343:390 In this study, HDL was isolated from plasma by sequential micro-ultracentrifugation, followed by glycoprotein and glycolipid analysis. 24417605 9 31 theme ceramide 1346:1353 arg1 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 31 theme ceramide 1346:1353 arg1 tails 1361:1365 heterogeneous ceramide lipid tails 1332:1365 heterogeneous ceramide lipid tails 1332:1365 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 31 theme ceramide 1346:1353 arg1 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 0 32 theme high 21:24 arg1 lipoprotein 34:44 high density lipoprotein 21:44 high density lipoprotein 21:44 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 6 33 theme abundant 730:737 arg1 N-glycan 739:746 The most abundant N-glycan 721:746 The most abundant N-glycan 721:746 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 33 theme abundant 730:737 arg1 type 774:777 a biantennary complex type glycan 752:784 a biantennary complex type glycan 752:784 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 8 34 theme GD3 1267:1269 arg1 distribution 1271:1282 A 60% GM3 and 40% GD3 distribution 1249:1282 A 60% GM3 and 40% GD3 distribution 1249:1282 A 60% GM3 and 40% GD3 distribution was observed. 24417605 4 35 theme HDL 560:562 arg1 particles 564:572 HDL particles 560:572 HDL particles 560:572 HDL particles were found to be highly sialylated. 24417605 1 36 theme functional 94:103 arg1 proteins 105:112 functional proteins 94:112 functional proteins 94:112 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 3 37 theme quadrupole 505:514 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 0 38 theme lipoprotein 34:44 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of high density lipoprotein 0:44 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 7 39 gly sialylated 948:957 arg1 O-glycans 929:937 The observed O-glycans 916:937 The observed O-glycans 916:937 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 10 40 theme first 1435:1439 arg1 time 1441:1444 the first time 1431:1444 the first time 1431:1444 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 7 41 theme major 1222:1226 arg1 GM3 1103:1105 fetuin A. GM3 1093:1105 fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) 1093:1151 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 41 theme major 1222:1226 arg1 gangliosides 1228:1239 the major gangliosides 1218:1239 the major gangliosides in HDL 1218:1246 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 41 theme major 1222:1226 arg1 CIII 1073:1076 apolipoprotein CIII 1058:1076 apolipoprotein CIII (ApoC-III) 1058:1087 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 8 42 theme % 1265:1265 arg1 GD3 1267:1269 40% GD3 1263:1269 40% GD3 1263:1269 A 60% GM3 and 40% GD3 distribution was observed. 24417605 0 43 theme density 26:32 arg1 lipoprotein 34:44 high density lipoprotein 21:44 high density lipoprotein 21:44 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 8 44 theme 40 1263:1264 arg1 % 1265:1265 % 1265:1265 % 1265:1265 A 60% GM3 and 40% GD3 distribution was observed. 24417605 6 45 with type 774:777 arg1 Hexose5HexNAc4Neu5Ac2 809:829 Hexose5HexNAc4Neu5Ac2 809:829 Hexose5HexNAc4Neu5Ac2 809:829 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 45 with type 774:777 arg1 acids 802:806 two sialic acids 791:806 two sialic acids (Hexose5HexNAc4Neu5Ac2) 791:830 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 7 46 theme core 981:984 arg1 structure 988:996 a core 1 structure 979:996 a core 1 structure 979:996 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 3 47 theme nano-HPLC 490:498 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 6 48 theme biantennary 754:764 arg1 N-glycan 739:746 The most abundant N-glycan 721:746 The most abundant N-glycan 721:746 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 48 theme biantennary 754:764 arg1 type 774:777 a biantennary complex type glycan 752:784 a biantennary complex type glycan 752:784 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 7 49 dep GM3 1103:1105 arg1 monosialoganglioside 1108:1127 monosialoganglioside 1108:1127 monosialoganglioside 1108:1127 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 49 dep GM3 1103:1105 arg1 NeuAc2-8NeuAc2-3Gal1-4Glc-Cer 1182:1210 NeuAc2-8NeuAc2-3Gal1-4Glc-Cer 1182:1210 NeuAc2-8NeuAc2-3Gal1-4Glc-Cer 1182:1210 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 49 dep GM3 1103:1105 arg1 disialoganglioside 1162:1179 disialoganglioside 1162:1179 disialoganglioside 1162:1179 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 49 dep GM3 1103:1105 arg1 NeuAc2-3Gal1-4Glc-Cer 1130:1150 NeuAc2-3Gal1-4Glc-Cer 1130:1150 NeuAc2-3Gal1-4Glc-Cer 1130:1150 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 10 50 gly sialylated 1518:1527 arg1 particles 1529:1537 highly sialylated particles 1511:1537 highly sialylated particles 1511:1537 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 10 50 gly sialylated 1518:1527 arg1 HDL 1503:1505 HDL 1503:1505 HDL 1503:1505 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 1 51 theme proteins 105:112 arg1 lipids 118:123 lipids 118:123 lipids 118:123 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 51 theme proteins 105:112 arg1 proteins 105:112 functional proteins 94:112 functional proteins 94:112 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 51 theme proteins 105:112 arg1 Many 82:85 Many 82:85 Many 82:85 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 5 52 gly glycoproteins 648:660 arg1 glycoproteins 648:660 HDL glycoproteins 644:660 HDL glycoproteins 644:660 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 5 53 theme HDL 644:646 arg1 glycoproteins 648:660 HDL glycoproteins 644:660 HDL glycoproteins 644:660 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 6 54 theme sialic 795:800 arg1 Hexose5HexNAc4Neu5Ac2 809:829 Hexose5HexNAc4Neu5Ac2 809:829 Hexose5HexNAc4Neu5Ac2 809:829 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 54 theme sialic 795:800 arg1 acids 802:806 two sialic acids 791:806 two sialic acids (Hexose5HexNAc4Neu5Ac2) 791:830 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 3 55 theme Chip 500:503 arg1 time 516:519 nano-HPLC Chip quadrupole time 490:519 nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 490:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 5 56 gly sialylated 667:676 arg1 Most 610:613 Most 610:613 Most 610:613 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 5 56 gly sialylated 667:676 arg1 N-glycans 622:630 the N-glycans 618:630 the N-glycans (∼90%) from HDL glycoproteins 618:660 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 5 56 gly sialylated 667:676 arg1 % 636:636 ∼90% 633:636 ∼90% 633:636 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 6 57 theme glycosylation 892:904 arg1 analysis 906:913 site-specific glycosylation analysis 878:913 site-specific glycosylation analysis 878:913 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 1 58 theme lipids 118:123 arg1 lipids 118:123 lipids 118:123 lipids 118:123 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 58 theme lipids 118:123 arg1 proteins 105:112 functional proteins 94:112 functional proteins 94:112 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 58 theme lipids 118:123 arg1 Many 82:85 Many 82:85 Many 82:85 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 5 59 from glycoproteins 648:660 arg1 N-glycans 622:630 the N-glycans 618:630 the N-glycans (∼90%) from HDL glycoproteins 618:660 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 5 59 from glycoproteins 648:660 arg1 % 636:636 ∼90% 633:636 ∼90% 633:636 Most of the N-glycans (∼90%) from HDL glycoproteins were sialylated with one or two neuraminic acids (Neu5Ac). 24417605 10 60 theme glycomic 1448:1455 arg1 approach 1457:1464 a glycomic approach 1446:1464 a glycomic approach for analyzing HDL 1446:1482 This report describes for the first time a glycomic approach for analyzing HDL, highlighting that HDL are highly sialylated particles. 24417605 3 61 dep extracted 441:449 arg1 followed 464:471 followed 464:471 followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS 464:557 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 9 62 theme heterogeneous 1332:1344 arg1 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 d18:1/23:0 1393:1402 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 62 theme heterogeneous 1332:1344 arg1 tails 1361:1365 heterogeneous ceramide lipid tails 1332:1365 heterogeneous ceramide lipid tails 1332:1365 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 9 62 theme heterogeneous 1332:1344 arg1 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 d18:1/16:0 1378:1387 Both GM3 and GD3 were composed of heterogeneous ceramide lipid tails, including d18:1/16:0 and d18:1/23:0. 24417605 8 63 theme GM3 1255:1257 arg1 distribution 1271:1282 A 60% GM3 and 40% GD3 distribution 1249:1282 A 60% GM3 and 40% GD3 distribution 1249:1282 A 60% GM3 and 40% GD3 distribution was observed. 24417605 0 64 theme sialylated 61:70 arg1 particle 72:79 a highly sialylated particle 52:79 a highly sialylated particle 52:79 Glycomic analysis of high density lipoprotein shows a highly sialylated particle. 24417605 6 65 gly glycoproteins 858:870 arg1 glycoproteins 858:870 multiple glycoproteins 849:870 multiple glycoproteins using site-specific glycosylation analysis 849:913 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 1 66 theme high 128:131 arg1 HDL 154:156 HDL 154:156 HDL 154:156 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 66 theme high 128:131 arg1 lipoprotein 141:151 high density lipoprotein 128:151 high density lipoprotein (HDL) particles 128:167 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 6 67 theme multiple 849:856 arg1 glycoproteins 858:870 multiple glycoproteins 849:870 multiple glycoproteins using site-specific glycosylation analysis 849:913 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 6 68 theme site-specific 878:890 arg1 analysis 906:913 site-specific glycosylation analysis 878:913 site-specific glycosylation analysis 878:913 The most abundant N-glycan was a biantennary complex type glycan with two sialic acids (Hexose5HexNAc4Neu5Ac2) and was found in multiple glycoproteins using site-specific glycosylation analysis. 24417605 3 69 gly glycopeptides 404:416 arg2 glycopeptides 404:416 glycopeptides 404:416 glycopeptides 404:416 N-Glycans, glycopeptides, and gangliosides were extracted and purified followed by analysis with nano-HPLC Chip quadrupole time of flight mass spectrometry and MS/MS. 24417605 1 70 theme density 133:139 arg1 HDL 154:156 HDL 154:156 HDL 154:156 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 1 70 theme density 133:139 arg1 lipoprotein 141:151 high density lipoprotein 128:151 high density lipoprotein (HDL) particles 128:167 Many of the functional proteins and lipids in high density lipoprotein (HDL) particles are potentially glycosylated, yet very little is known about the glycoconjugates of HDL. 24417605 8 71 theme 60 1251:1252 arg1 % 1253:1253 % 1253:1253 % 1253:1253 A 60% GM3 and 40% GD3 distribution was observed. 24417605 7 72 contain contained 969:977 arg2 structure 988:996 a core 1 structure 979:996 a core 1 structure 979:996 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 24417605 7 72 contain contained 969:977 arg1 most 964:967 most 964:967 most 964:967 The observed O-glycans were all sialylated, and most contained a core 1 structure with two Neu5Acs, including those that were associated with apolipoprotein CIII (ApoC-III) and fetuin A. GM3 (monosialoganglioside, NeuAc2-3Gal1-4Glc-Cer) and GD3 (disialoganglioside, NeuAc2-8NeuAc2-3Gal1-4Glc-Cer) were the major gangliosides in HDL. 27913739 8 0 theme -COOH 1181:1185 arg1 Removal 1150:1156 Removal 1150:1156 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide 1150:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 10 1 theme sugar 1538:1542 arg1 analogs 1544:1550 the sugar analogs 1534:1550 the sugar analogs tested for pollen germination 1534:1580 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 10 2 theme 4-Me-GlcA-β 1429:1439 arg1 -Gal 1445:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 11 3 theme sugar 1677:1681 arg1 molecule 1683:1690 a sugar molecule 1675:1690 a sugar molecule involved in plant reproduction 1675:1721 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 3 4 theme tubes 537:541 arg1 response 518:525 the response 514:525 the response of pollen tubes to these ovular attractants 514:569 Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. 27913739 6 5 theme noncritical 918:928 arg1 structures 930:939 critical and noncritical structures 905:939 critical and noncritical structures in the disaccharide 905:959 However, critical and noncritical structures in the disaccharide have not been dissected deeply. 27913739 10 6 theme pollen 1563:1568 arg1 germination 1570:1580 pollen germination 1563:1580 pollen germination 1563:1580 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 7 7 theme synthesized 1129:1139 arg1 analogs 1141:1147 these synthesized analogs 1123:1147 these synthesized analogs 1123:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 10 8 theme -Gal 1445:1448 arg1 specific 1458:1465 specific 1458:1465 specific 1458:1465 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 10 8 theme -Gal 1445:1448 arg1 role 1389:1392 The role 1385:1392 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal 1385:1448 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 6 9 theme critical 905:912 arg1 structures 930:939 critical and noncritical structures 905:939 critical and noncritical structures in the disaccharide 905:959 However, critical and noncritical structures in the disaccharide have not been dissected deeply. 27913739 4 10 theme pollen 678:683 arg1 tube 685:688 the pollen tube 674:688 the pollen tube 674:688 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 10 11 theme 1,6 1441:1443 arg1 -Gal 1445:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 1 12 theme tube 205:208 arg1 control 187:193 the precise directional growth control 156:193 the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 156:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 8 13 theme AMOR 1258:1261 arg1 activity 1263:1270 AMOR activity 1258:1270 AMOR activity 1258:1270 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 7 14 theme analogs 1037:1043 arg1 synthesis 1015:1023 the synthesis 1011:1023 the synthesis of new AMOR analogs 1011:1043 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 7 14 theme analogs 1037:1043 arg1 relationships 1072:1084 the structure-activity relationships 1049:1084 the structure-activity relationships for AMOR activity in the presence of these synthesized analogs 1049:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 1 15 theme pistil 229:234 arg1 tissue 236:241 the female pistil tissue 218:241 the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 218:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 4 16 theme attractant 711:720 arg1 peptides 727:734 ovular attractant LURE peptides 704:734 ovular attractant LURE peptides in Torenia fournieri 704:755 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 7 17 theme new 1028:1030 arg1 analogs 1037:1043 new AMOR analogs 1028:1043 new AMOR analogs 1028:1043 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 7 18 theme structure-activity 1053:1070 arg1 relationships 1072:1084 the structure-activity relationships 1049:1084 the structure-activity relationships for AMOR activity in the presence of these synthesized analogs 1049:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 9 19 theme sugar 1309:1313 arg1 backbone 1286:1293 The pyranose backbone 1273:1293 The pyranose backbone of the second sugar of disaccharide 1273:1329 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 9 19 theme sugar 1309:1313 arg1 essential 1334:1342 essential 1334:1342 essential 1334:1342 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 5 20 theme beta 762:765 arg1 essential 841:849 essential 841:849 essential 841:849 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 5 20 theme beta 762:765 arg1 isomer 767:772 The beta isomer 758:772 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose 758:835 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 9 21 theme disaccharide 1318:1329 arg1 sugar 1309:1313 the second sugar 1298:1313 the second sugar of disaccharide 1298:1329 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 1 22 theme precise 160:166 arg1 control 187:193 the precise directional growth control 156:193 the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 156:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 0 23 theme Ovular 83:88 arg1 Guidance 90:97 Ovular Guidance 83:97 Ovular Guidance 83:97 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 1 24 theme growth 180:185 arg1 control 187:193 the precise directional growth control 156:193 the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 156:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 2 25 theme cell 418:421 arg1 side 402:405 either side 395:405 either side of the egg cell 395:421 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 0 26 theme Structure-Activity 0:17 arg1 Relation 19:26 Structure-Activity Relation 0:26 Structure-Activity Relation of AMOR Sugar Molecule That 0:54 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 9 27 theme hydroxy 1369:1375 arg1 groups 1377:1382 hydroxy groups 1369:1382 the activity but not hydroxy groups 1348:1382 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 1 28 theme Successful 100:109 arg1 fertilization 111:123 Successful fertilization 100:123 Successful fertilization in flowering plants 100:143 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 2 29 theme ovular 430:435 arg1 attractants 437:447 ovular attractants 430:447 ovular attractants of pollen tubes 430:463 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 2 29 theme ovular 430:435 arg1 peptides 344:351 Cys-rich peptides 335:351 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell 335:421 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 2 30 theme egg 414:416 arg1 cell 418:421 the egg cell 410:421 the egg cell 410:421 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 0 31 theme Sugar 36:40 arg1 Molecule 42:49 AMOR Sugar Molecule 31:49 AMOR Sugar Molecule That 31:54 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 1 32 theme nonmotile 312:320 arg1 cells 328:332 nonmotile sperm cells 312:332 nonmotile sperm cells 312:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 5 33 theme terminal 781:788 arg1 galactose 827:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 2 34 theme tubes 459:463 arg1 attractants 437:447 ovular attractants 430:447 ovular attractants of pollen tubes 430:463 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 2 34 theme tubes 459:463 arg1 peptides 344:351 Cys-rich peptides 335:351 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell 335:421 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 11 35 theme first 1609:1613 arg1 relationship 1634:1645 the first structure-activity relationship 1605:1645 the first structure-activity relationship study 1605:1651 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 5 36 theme 4-O-methyl-glucuronosyl 803:825 arg1 galactose 827:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 1 37 theme cells 328:332 arg1 delivery 300:307 delivery 300:307 delivery of nonmotile sperm cells 300:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 8 38 from residue 1209:1215 arg1 Removal 1150:1156 Removal 1150:1156 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide 1150:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 10 39 theme beta 1397:1400 arg1 isomer 1402:1407 beta isomer 1397:1407 beta isomer 1397:1407 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 8 40 theme 4-O-methyl 1161:1170 arg1 group 1172:1176 4-O-methyl group 1161:1176 4-O-methyl group 1161:1176 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 4 41 theme 4-O-methyl-glucuronosyl 605:627 arg1 AMOR 646:649 AMOR 646:649 AMOR 646:649 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 4 41 theme 4-O-methyl-glucuronosyl 605:627 arg1 arabinogalactan 629:643 ovular 4-O-methyl-glucuronosyl arabinogalactan 598:643 ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) 598:650 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 11 42 theme relationship 1634:1645 arg1 study 1647:1651 the first structure-activity relationship study 1605:1651 the first structure-activity relationship study 1605:1651 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 7 43 from activity 1095:1102 arg1 presence 1111:1118 the presence 1107:1118 the presence of these synthesized analogs 1107:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 7 44 theme analogs 1141:1147 arg1 presence 1111:1118 the presence 1107:1118 the presence of these synthesized analogs 1107:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 10 45 theme disaccharide 1416:1427 arg1 -Gal 1445:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 the disaccharide 4-Me-GlcA-β(1,6)-Gal 1412:1448 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 3 46 theme pollen 530:535 arg1 tubes 537:541 pollen tubes 530:541 pollen tubes 530:541 Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. 27913739 6 47 from structures 930:939 arg1 disaccharide 948:959 the disaccharide 944:959 the disaccharide 944:959 However, critical and noncritical structures in the disaccharide have not been dissected deeply. 27913739 11 48 theme molecule 1683:1690 arg1 study 1647:1651 the first structure-activity relationship study 1605:1651 the first structure-activity relationship study 1605:1651 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 8 49 theme glucuronosyl 1196:1207 arg1 disaccharide 1224:1235 the disaccharide 1220:1235 the disaccharide 1220:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 8 49 theme glucuronosyl 1196:1207 arg1 residue 1209:1215 the glucuronosyl residue 1192:1215 the glucuronosyl residue of the disaccharide 1192:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 3 50 theme ovular 552:557 arg1 attractants 559:569 these ovular attractants 546:569 these ovular attractants 546:569 Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. 27913739 1 51 theme pollen 198:203 arg1 tube 205:208 pollen tube 198:208 pollen tube 198:208 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 8 52 theme disaccharide 1224:1235 arg1 disaccharide 1224:1235 the disaccharide 1220:1235 the disaccharide 1220:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 8 52 theme disaccharide 1224:1235 arg1 residue 1209:1215 the glucuronosyl residue 1192:1215 the glucuronosyl residue of the disaccharide 1192:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 11 53 theme activity 1791:1798 arg1 loss 1783:1786 loss 1783:1786 loss of activity 1783:1798 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 7 54 theme AMOR 1090:1093 arg1 activity 1095:1102 AMOR activity 1090:1102 AMOR activity in the presence of these synthesized analogs 1090:1147 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 9 55 theme pyranose 1277:1284 arg1 backbone 1286:1293 The pyranose backbone 1273:1293 The pyranose backbone of the second sugar of disaccharide 1273:1329 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 9 55 theme pyranose 1277:1284 arg1 essential 1334:1342 essential 1334:1342 essential 1334:1342 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 1 56 theme female 222:227 arg1 tissue 236:241 the female pistil tissue 218:241 the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 218:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 4 57 from peptides 727:734 arg1 fournieri 747:755 fournieri 747:755 fournieri 747:755 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 1 58 theme flowering 128:136 arg1 plants 138:143 flowering plants 128:143 flowering plants 128:143 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 4 59 theme ovular 704:709 arg1 peptides 727:734 ovular attractant LURE peptides 704:734 ovular attractant LURE peptides in Torenia fournieri 704:755 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 9 60 theme second 1302:1307 arg1 sugar 1309:1313 the second sugar 1298:1313 the second sugar of disaccharide 1298:1329 The pyranose backbone of the second sugar of disaccharide is essential for the activity but not hydroxy groups. 27913739 7 61 theme AMOR 1032:1035 arg1 analogs 1037:1043 new AMOR analogs 1028:1043 new AMOR analogs 1028:1043 Herein, we report the synthesis of new AMOR analogs and the structure-activity relationships for AMOR activity in the presence of these synthesized analogs. 27913739 4 62 theme LURE 722:725 arg1 peptides 727:734 ovular attractant LURE peptides 704:734 ovular attractant LURE peptides in Torenia fournieri 704:755 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 10 63 from difference 1507:1516 arg1 effect 1521:1526 effect 1521:1526 effect 1521:1526 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 4 64 theme tube 685:688 arg1 competency 660:669 competency 660:669 competency of the pollen tube 660:688 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 1 65 theme directional 168:178 arg1 control 187:193 the precise directional growth control 156:193 the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 156:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 11 66 theme molecule 1766:1773 arg1 modification 1746:1757 modification 1746:1757 modification of the molecule 1746:1773 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 5 67 theme galactose 827:835 arg1 essential 841:849 essential 841:849 essential 841:849 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 5 67 theme galactose 827:835 arg1 isomer 767:772 The beta isomer 758:772 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose 758:835 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 10 68 theme competency 1471:1480 arg1 control 1482:1488 competency control 1471:1488 competency control 1471:1488 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 0 69 theme AMOR 31:34 arg1 Molecule 42:49 AMOR Sugar Molecule 31:49 AMOR Sugar Molecule That 31:54 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 11 70 theme plant 1704:1708 arg1 reproduction 1710:1721 plant reproduction 1704:1721 plant reproduction 1704:1721 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 3 71 theme Competency 466:475 arg1 control 477:483 Competency control 466:483 Competency control by the pistil 466:497 Competency control by the pistil is crucial for the response of pollen tubes to these ovular attractants. 27913739 0 72 theme Molecule 42:49 arg1 Relation 19:26 Structure-Activity Relation 0:26 Structure-Activity Relation of AMOR Sugar Molecule That 0:54 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 5 73 theme disaccharide 790:801 arg1 galactose 827:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 the terminal disaccharide 4-O-methyl-glucuronosyl galactose 777:835 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 1 74 theme sperm 322:326 arg1 cells 328:332 nonmotile sperm cells 312:332 nonmotile sperm cells 312:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 0 75 dep Molecule 42:49 arg1 That 51:54 That 51:54 That 51:54 Structure-Activity Relation of AMOR Sugar Molecule That Activates Pollen-Tubes for Ovular Guidance. 27913739 2 76 theme pollen 452:457 arg1 tubes 459:463 pollen tubes 452:463 pollen tubes 452:463 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 1 77 from fertilization 111:123 arg1 plants 138:143 flowering plants 128:143 flowering plants 128:143 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 27913739 4 78 theme ovular 598:603 arg1 AMOR 646:649 AMOR 646:649 AMOR 646:649 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 4 78 theme ovular 598:603 arg1 arabinogalactan 629:643 ovular 4-O-methyl-glucuronosyl arabinogalactan 598:643 ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) 598:650 We recently reported that ovular 4-O-methyl-glucuronosyl arabinogalactan (AMOR) induces competency of the pollen tube to respond to ovular attractant LURE peptides in Torenia fournieri. 27913739 10 79 theme isomer 1402:1407 arg1 specific 1458:1465 specific 1458:1465 specific 1458:1465 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 10 79 theme isomer 1402:1407 arg1 role 1389:1392 The role 1385:1392 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal 1385:1448 The role of beta isomer of the disaccharide 4-Me-GlcA-β(1,6)-Gal is very specific for competency control, as there was no difference in effect among the sugar analogs tested for pollen germination. 27913739 2 80 theme Cys-rich 335:342 arg1 attractants 437:447 ovular attractants 430:447 ovular attractants of pollen tubes 430:463 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 2 80 theme Cys-rich 335:342 arg1 peptides 344:351 Cys-rich peptides 335:351 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell 335:421 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 8 81 theme group 1172:1176 arg1 Removal 1150:1156 Removal 1150:1156 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide 1150:1235 Removal of 4-O-methyl group or -COOH from the glucuronosyl residue of the disaccharide dramatically reduces AMOR activity. 27913739 5 82 theme competency 874:883 arg1 induction 885:893 the competency induction 870:893 the competency induction 870:893 The beta isomer of the terminal disaccharide 4-O-methyl-glucuronosyl galactose was essential and sufficient for the competency induction. 27913739 11 83 theme structure-activity 1615:1632 arg1 relationship 1634:1645 the first structure-activity relationship 1605:1645 the first structure-activity relationship study 1605:1651 This study represents the first structure-activity relationship study, to our knowledge, of a sugar molecule involved in plant reproduction, which opens a way for modification of the molecule without loss of activity. 27913739 2 84 theme synergid 377:384 arg1 cells 386:390 the synergid cells 373:390 the synergid cells 373:390 Cys-rich peptides LUREs secreted from the synergid cells on either side of the egg cell act as ovular attractants of pollen tubes. 27913739 1 85 theme female 254:259 arg1 gametophyte 261:271 the female gametophyte 250:271 the female gametophyte contained in the ovule for delivery of nonmotile sperm cells 250:332 Successful fertilization in flowering plants depends on the precise directional growth control of pollen tube through the female pistil tissue toward the female gametophyte contained in the ovule for delivery of nonmotile sperm cells. 28334636 0 0 theme α-glucosyl 89:98 arg1 rutin 100:104 α-glucosyl rutin 89:104 α-glucosyl rutin 89:104 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 6 1 theme transglycosylated 1001:1017 arg1 compounds 1019:1027 transglycosylated compounds 1001:1027 transglycosylated compounds in food, cosmetic, and pharmaceutical fields 1001:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 6 2 from compounds 1019:1027 arg1 cosmetic 1038:1045 cosmetic 1038:1045 cosmetic 1038:1045 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 6 2 from compounds 1019:1027 arg1 food 1032:1035 food 1032:1035 food 1032:1035 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 6 2 from compounds 1019:1027 arg1 fields 1067:1072 pharmaceutical fields 1052:1072 pharmaceutical fields 1052:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 1 3 theme 1H 160:161 arg1 signals 175:181 1H and 13C NMR signals 160:181 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 160:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 4 theme novel 228:232 arg1 rhoifolin 207:215 α-glucosyl rhoifolin 196:215 α-glucosyl rhoifolin (Rhf-G) 196:223 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 4 theme novel 228:232 arg1 compound 252:259 a novel transglycosylated compound 226:259 a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 226:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 3 5 theme chemical 679:686 arg1 structure 688:696 its chemical structure 675:696 its chemical structure 675:696 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 1 6 theme transglycosylated 234:250 arg1 rhoifolin 207:215 α-glucosyl rhoifolin 196:215 α-glucosyl rhoifolin (Rhf-G) 196:223 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 6 theme transglycosylated 234:250 arg1 compound 252:259 a novel transglycosylated compound 226:259 a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 226:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 7 theme chemical 325:332 arg1 structure 334:342 its chemical structure 321:342 a flavone glycoside, rhoifolin, as well as its chemical structure 278:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 6 8 gly transglycosylated 1001:1017 arg1 compounds 1019:1027 transglycosylated compounds 1001:1027 transglycosylated compounds in food, cosmetic, and pharmaceutical fields 1001:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 6 9 theme NMR 986:988 arg1 studies 990:996 comprehensive NMR studies 972:996 comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields 972:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 6 10 from studies 990:996 arg1 compounds 1019:1027 transglycosylated compounds 1001:1027 transglycosylated compounds in food, cosmetic, and pharmaceutical fields 1001:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 5 11 theme intramolecular 830:843 arg1 bonds 854:858 intramolecular hydrogen bonds 830:858 intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons 830:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 5 12 theme basic 867:871 arg1 skeletons 891:899 the basic Rhf-G and Rutin-G skeletons 863:899 the basic Rhf-G and Rutin-G skeletons 863:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 0 13 theme NMR 109:111 arg1 spectroscopy 113:124 NMR spectroscopy 109:124 NMR spectroscopy 109:124 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 2 14 theme transglycosylated 415:431 arg1 compound 433:440 another transglycosylated compound 407:440 another transglycosylated compound 407:440 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 2 14 theme transglycosylated 415:431 arg1 rutin 454:458 α-glucosyl rutin 443:458 α-glucosyl rutin (Rutin-G) 443:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 1 15 theme 13C 167:169 arg1 NMR 171:173 13C NMR 167:173 13C NMR 167:173 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 16 theme NMR 171:173 arg1 signals 175:181 1H and 13C NMR signals 160:181 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 160:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 0 17 theme Structural 0:9 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of a novel 0:32 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 6 18 theme comprehensive 972:984 arg1 studies 990:996 comprehensive NMR studies 972:996 comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields 972:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 1 19 theme signals 175:181 arg1 assignment 146:155 the full assignment 137:155 the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 137:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 5 20 theme hydrogen 845:852 arg1 bonds 854:858 intramolecular hydrogen bonds 830:858 intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons 830:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 1 21 gly transglycosylated 234:250 arg1 rhoifolin 207:215 α-glucosyl rhoifolin 196:215 α-glucosyl rhoifolin (Rhf-G) 196:223 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 21 gly transglycosylated 234:250 arg1 compound 252:259 a novel transglycosylated compound 226:259 a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 226:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 6 22 theme pharmaceutical 1052:1065 arg1 fields 1067:1072 pharmaceutical fields 1052:1072 pharmaceutical fields 1052:1072 These findings will be helpful for comprehensive NMR studies on transglycosylated compounds in food, cosmetic, and pharmaceutical fields. 28334636 2 23 theme signal 385:390 arg1 assignment 392:401 the complete NMR signal assignment 368:401 the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G) 368:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 1 24 theme flavone 280:286 arg1 rhoifolin 299:307 rhoifolin 299:307 rhoifolin 299:307 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 24 theme flavone 280:286 arg1 glycoside 288:296 a flavone glycoside 278:296 a flavone glycoside 278:296 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 2 25 theme sugar 507:511 arg1 moieties 513:520 its sugar moieties 503:520 its sugar moieties 503:520 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 1 26 theme α-glucosyl 196:205 arg1 compound 252:259 a novel transglycosylated compound 226:259 a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 226:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 26 theme α-glucosyl 196:205 arg1 Rhf-G 218:222 Rhf-G 218:222 Rhf-G 218:222 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 1 26 theme α-glucosyl 196:205 arg1 rhoifolin 207:215 α-glucosyl rhoifolin 196:215 α-glucosyl rhoifolin (Rhf-G) 196:223 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 4 27 theme moiety 807:812 arg1 position 782:789 position 782:789 position 3a of the sugar moiety 782:812 The additional glucose was bound directly via a transglycosylation to rhoifolin at position 3a of the sugar moiety. 28334636 3 28 dep along 589:593 arg1 with 595:598 with 595:598 with 595:598 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 2 29 theme NMR 381:383 arg1 assignment 392:401 the complete NMR signal assignment 368:401 the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G) 368:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 0 30 theme novel 28:32 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of a novel 0:32 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 4 31 theme sugar 801:805 arg1 moiety 807:812 the sugar moiety 797:812 the sugar moiety 797:812 The additional glucose was bound directly via a transglycosylation to rhoifolin at position 3a of the sugar moiety. 28334636 2 32 theme complete 372:379 arg1 assignment 392:401 the complete NMR signal assignment 368:401 the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G) 368:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 0 33 theme α-glucosyl 61:70 arg1 rhoifolin 72:80 compound α-glucosyl rhoifolin 52:80 compound α-glucosyl rhoifolin 52:80 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 3 34 theme Electrospray 547:558 arg1 spectrometry 576:587 Electrospray ionization-mass spectrometry 547:587 Electrospray ionization-mass spectrometry along with multiple NMR methods 547:619 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 5 35 from bonds 854:858 arg1 skeletons 891:899 the basic Rhf-G and Rutin-G skeletons 863:899 the basic Rhf-G and Rutin-G skeletons 863:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 4 36 theme additional 703:712 arg1 glucose 714:720 The additional glucose 699:720 The additional glucose 699:720 The additional glucose was bound directly via a transglycosylation to rhoifolin at position 3a of the sugar moiety. 28334636 0 37 theme compound 52:59 arg1 rhoifolin 72:80 compound α-glucosyl rhoifolin 52:80 compound α-glucosyl rhoifolin 52:80 Structural elucidation of a novel transglycosylated compound α-glucosyl rhoifolin and of α-glucosyl rutin by NMR spectroscopy. 28334636 3 38 theme ionization-mass 560:574 arg1 spectrometry 576:587 Electrospray ionization-mass spectrometry 547:587 Electrospray ionization-mass spectrometry along with multiple NMR methods 547:619 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 3 39 theme NMR 609:611 arg1 methods 613:619 multiple NMR methods 600:619 Electrospray ionization-mass spectrometry along with multiple NMR methods 547:619 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 2 40 gly transglycosylated 415:431 arg1 compound 433:440 another transglycosylated compound 407:440 another transglycosylated compound 407:440 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 2 40 gly transglycosylated 415:431 arg1 rutin 454:458 α-glucosyl rutin 443:458 α-glucosyl rutin (Rutin-G) 443:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 5 41 theme HMBC 919:922 arg1 experiments 924:934 HMBC experiments 919:934 HMBC experiments 919:934 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 5 42 theme Rhf-G 873:877 arg1 skeletons 891:899 the basic Rhf-G and Rutin-G skeletons 863:899 the basic Rhf-G and Rutin-G skeletons 863:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 3 43 theme sugar 657:661 arg1 moieties 663:670 three sugar moieties 651:670 three sugar moieties 651:670 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 3 44 contain possesses 641:649 arg1 Rhf-G 635:639 Rhf-G 635:639 Rhf-G 635:639 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 3 44 contain possesses 641:649 arg2 moieties 663:670 three sugar moieties 651:670 three sugar moieties 651:670 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 1 45 theme full 141:144 arg1 assignment 146:155 the full assignment 137:155 the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure 137:342 We report the full assignment of 1H and 13C NMR signals belonging to α-glucosyl rhoifolin (Rhf-G), a novel transglycosylated compound synthesized from a flavone glycoside, rhoifolin, as well as its chemical structure. 28334636 2 46 theme α-glucosyl 443:452 arg1 Rutin-G 461:467 Rutin-G 461:467 Rutin-G 461:467 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 2 46 theme α-glucosyl 443:452 arg1 compound 433:440 another transglycosylated compound 407:440 another transglycosylated compound 407:440 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 2 46 theme α-glucosyl 443:452 arg1 rutin 454:458 α-glucosyl rutin 443:458 α-glucosyl rutin (Rutin-G) 443:468 Furthermore, we report the complete NMR signal assignment for another transglycosylated compound, α-glucosyl rutin (Rutin-G), as the signals corresponding to its sugar moieties had not been identified. 28334636 3 47 theme multiple 600:607 arg1 methods 613:619 multiple NMR methods 600:619 Electrospray ionization-mass spectrometry along with multiple NMR methods 547:619 Electrospray ionization-mass spectrometry along with multiple NMR methods revealed that Rhf-G possesses three sugar moieties in its chemical structure. 28334636 5 48 theme Rutin-G 883:889 arg1 skeletons 891:899 the basic Rhf-G and Rutin-G skeletons 863:899 the basic Rhf-G and Rutin-G skeletons 863:899 Interestingly, intramolecular hydrogen bonds in the basic Rhf-G and Rutin-G skeletons were confirmed by HMBC experiments. 28334636 4 49 dep position 782:789 arg1 3a 791:792 3a 791:792 position 3a of the sugar moiety 782:812 The additional glucose was bound directly via a transglycosylation to rhoifolin at position 3a of the sugar moiety. 25269761 10 0 theme β3GnT8 1247:1252 arg1 expression 1254:1263 β3GnT8 expression 1247:1263 β3GnT8 expression 1247:1263 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 11 1 theme treatment 1637:1645 arg1 strategies 1647:1656 new treatment strategies 1633:1656 new treatment strategies 1633:1656 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 7 2 theme western 886:892 arg1 analysis 899:906 Quantitative RT-‑PCR and western blot analysis 861:906 analysis 899:906 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 8 3 theme SW620/5-FU 1089:1098 arg1 cells 1100:1104 SW620/5-FU cells 1089:1104 SW620/5-FU cells 1089:1104 We found polylactosamine chains were significantly increased in SW620/5-FU cells. 25269761 11 4 theme chemotherapeutic 1556:1571 arg1 drug 1573:1576 the chemotherapeutic drug 1552:1576 the chemotherapeutic drug 1552:1576 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 10 5 theme polylactosamine 1445:1459 arg1 formation 1432:1440 the formation 1428:1440 the formation of polylactosamine 1428:1459 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 7 6 theme Quantitative 861:872 arg1 RT-‑PCR 874:880 Quantitative RT-‑PCR and western blot analysis 861:906 RT-‑PCR 874:880 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 3 7 theme 5-Fluorouracil 400:413 arg1 resistance 422:431 5-Fluorouracil (5-FU) resistance 400:431 5-Fluorouracil (5-FU) resistance 400:431 5-Fluorouracil (5-FU) resistance remains a major obstacle to the chemotherapy of CRC. 25269761 9 8 theme 5-FU 1203:1206 arg1 tolerance 1208:1216 5-FU tolerance 1203:1216 5-FU tolerance 1203:1216 Inhibition of the biosynthesis of polylactosamine by 3'-azidothymidine (AZT) was able to reduce 5-FU tolerance. 25269761 10 9 theme Further 1219:1225 arg1 studies 1227:1233 Further studies 1219:1233 Further studies 1219:1233 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 5 10 theme 5-FU 716:719 arg1 IC50 707:710 IC50 707:710 IC50 707:710 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 10 theme 5-FU 716:719 arg1 concentration 692:704 50% inhibition concentration 677:704 50% inhibition concentration (IC50) of 5-FU 677:719 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 8 11 theme polylactosamine 1034:1048 arg1 chains 1050:1055 polylactosamine chains 1034:1055 polylactosamine chains 1034:1055 We found polylactosamine chains were significantly increased in SW620/5-FU cells. 25269761 7 12 theme candidate 943:951 arg1 genes 953:957 candidate genes 943:957 candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells 943:1022 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 11 13 theme significant 1586:1596 arg1 implication 1598:1608 significant implication 1586:1608 significant implication for the development of new treatment strategies 1586:1656 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 2 14 theme colorectal 375:384 arg1 CRC 394:396 CRC 394:396 CRC 394:396 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8, which synthesizes polylactosamine on β1-6 branched N-glycans, is dramatically upregulated in colorectal cancer (CRC). 25269761 2 14 theme colorectal 375:384 arg1 cancer 386:391 colorectal cancer 375:391 colorectal cancer (CRC) 375:397 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8, which synthesizes polylactosamine on β1-6 branched N-glycans, is dramatically upregulated in colorectal cancer (CRC). 25269761 5 15 theme cell 635:638 arg1 SW620/5-FU 646:655 SW620/5-FU 646:655 SW620/5-FU 646:655 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 15 theme cell 635:638 arg1 line 640:643 a 5-FU‑resistant cell line 618:643 a 5-FU‑resistant cell line (SW620/5-FU) 618:656 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 16 theme 5-FU‑resistant 620:633 arg1 SW620/5-FU 646:655 SW620/5-FU 646:655 SW620/5-FU 646:655 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 16 theme 5-FU‑resistant 620:633 arg1 line 640:643 a 5-FU‑resistant cell line 618:643 a 5-FU‑resistant cell line (SW620/5-FU) 618:656 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 2 17 theme branched 324:331 arg1 N-glycans 333:341 β1-6 branched N-glycans 319:341 β1-6 branched N-glycans 319:341 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8, which synthesizes polylactosamine on β1-6 branched N-glycans, is dramatically upregulated in colorectal cancer (CRC). 25269761 7 18 theme blot 894:897 arg1 analysis 899:906 Quantitative RT-‑PCR and western blot analysis 861:906 analysis 899:906 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 0 19 from resistance 44:53 arg1 cells 82:86 human colorectal cancer cells 58:86 human colorectal cancer cells 58:86 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 11 20 theme tumor 1531:1535 arg1 sensitivity 1537:1547 tumor sensitivity 1531:1547 tumor sensitivity to the chemotherapeutic drug 1531:1576 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 6 21 theme lectin 769:774 arg1 analysis 781:788 lectin blot analysis 769:788 lectin blot analysis 769:788 Flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. 25269761 2 22 theme β1-6 319:322 arg1 N-glycans 333:341 β1-6 branched N-glycans 319:341 β1-6 branched N-glycans 319:341 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8, which synthesizes polylactosamine on β1-6 branched N-glycans, is dramatically upregulated in colorectal cancer (CRC). 25269761 3 23 theme CRC 481:483 arg1 chemotherapy 465:476 the chemotherapy 461:476 the chemotherapy of CRC 461:483 5-Fluorouracil (5-FU) resistance remains a major obstacle to the chemotherapy of CRC. 25269761 10 24 theme cancer 1304:1309 arg1 cells 1311:1315 5-FU‑resistant cancer cells 1289:1315 5-FU‑resistant cancer cells 1289:1315 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 1 25 theme cancer 209:214 arg1 chemoresistance 216:230 cancer chemoresistance 209:230 cancer chemoresistance 209:230 Aberrant glycosylation is known to be associated with cancer chemoresistance. 25269761 0 26 theme N-glycans 144:152 arg1 biosynthesis 107:118 the biosynthesis 103:118 the biosynthesis of polylactosamine-type N-glycans 103:152 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 6 27 theme Flow 750:753 arg1 cytometry 755:763 Flow cytometry 750:763 Flow cytometry 750:763 Flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. 25269761 4 28 from resistance 555:564 arg1 CRC 598:600 CRC 598:600 CRC 598:600 However, little is known with regard to the correlation between 5‑FU resistance and the expression of β3GnT8 in CRC. 25269761 10 29 theme β3GnT8 1335:1340 arg1 knockdown 1322:1330 knockdown 1322:1330 knockdown of β3GnT8 by RNA interference 1322:1360 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 4 30 from expression 574:583 arg1 CRC 598:600 CRC 598:600 CRC 598:600 However, little is known with regard to the correlation between 5‑FU resistance and the expression of β3GnT8 in CRC. 25269761 0 31 theme polylactosamine-type 123:142 arg1 N-glycans 144:152 polylactosamine-type N-glycans 123:152 polylactosamine-type N-glycans 123:152 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 9 32 theme biosynthesis 1125:1136 arg1 Inhibition 1107:1116 Inhibition 1107:1116 Inhibition of the biosynthesis of polylactosamine by 3'-azidothymidine (AZT) 1107:1182 Inhibition of the biosynthesis of polylactosamine by 3'-azidothymidine (AZT) was able to reduce 5-FU tolerance. 25269761 4 33 theme β3GnT8 588:593 arg1 resistance 555:564 5‑FU resistance 550:564 5‑FU resistance 550:564 However, little is known with regard to the correlation between 5‑FU resistance and the expression of β3GnT8 in CRC. 25269761 4 33 theme β3GnT8 588:593 arg1 expression 574:583 the expression 570:583 the expression of β3GnT8 in CRC 570:600 However, little is known with regard to the correlation between 5‑FU resistance and the expression of β3GnT8 in CRC. 25269761 6 34 theme structures 849:858 arg1 alteration 819:828 the alteration 815:828 the alteration of polylactosamine structures 815:858 Flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. 25269761 0 35 theme 5-fluorouracil 29:42 arg1 resistance 44:53 5-fluorouracil resistance 29:53 5-fluorouracil resistance in human colorectal cancer cells 29:86 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 6 36 theme polylactosamine 833:847 arg1 structures 849:858 polylactosamine structures 833:858 polylactosamine structures 833:858 Flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. 25269761 9 37 theme polylactosamine 1141:1155 arg1 biosynthesis 1125:1136 the biosynthesis 1121:1136 the biosynthesis of polylactosamine 1121:1155 Inhibition of the biosynthesis of polylactosamine by 3'-azidothymidine (AZT) was able to reduce 5-FU tolerance. 25269761 0 38 theme human 58:62 arg1 cells 82:86 human colorectal cancer cells 58:86 human colorectal cancer cells 58:86 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 4 39 theme 5‑FU 550:553 arg1 resistance 555:564 5‑FU resistance 550:564 5‑FU resistance 550:564 However, little is known with regard to the correlation between 5‑FU resistance and the expression of β3GnT8 in CRC. 25269761 5 40 theme MTT 739:741 arg1 assay 743:747 MTT assay 739:747 MTT assay 739:747 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 11 41 from alteration 1481:1490 arg1 cells 1509:1513 CRC cells 1505:1513 CRC cells 1505:1513 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 3 42 theme major 443:447 arg1 obstacle 449:456 a major obstacle 441:456 a major obstacle to the chemotherapy of CRC 441:483 5-Fluorouracil (5-FU) resistance remains a major obstacle to the chemotherapy of CRC. 25269761 11 43 theme β3GnT8 1495:1500 arg1 alteration 1481:1490 the alteration 1477:1490 the alteration of β3GnT8 in CRC cells 1477:1513 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 5 44 theme 50 677:678 arg1 % 679:679 % 679:679 % 679:679 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 7 45 used used 913:916 arg2 analysis 899:906 Quantitative RT-‑PCR and western blot analysis 861:906 analysis 899:906 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 7 45 used used 913:916 arg2 RT-‑PCR 874:880 Quantitative RT-‑PCR and western blot analysis 861:906 RT-‑PCR 874:880 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 11 46 theme CRC 1505:1507 arg1 cells 1509:1513 CRC cells 1505:1513 CRC cells 1505:1513 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 5 47 theme % 679:679 arg1 IC50 707:710 IC50 707:710 IC50 707:710 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 47 theme % 679:679 arg1 concentration 692:704 50% inhibition concentration 677:704 50% inhibition concentration (IC50) of 5-FU 677:719 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 2 48 theme β-1,3-N-acetyl-glucosaminyltransferase 233:270 arg1 β3GnT 273:277 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8 233:279 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8 233:279 β-1,3-N-acetyl-glucosaminyltransferase (β3GnT)8, which synthesizes polylactosamine on β1-6 branched N-glycans, is dramatically upregulated in colorectal cancer (CRC). 25269761 0 49 dep reverses 20:27 arg1 biosynthesis 107:118 the biosynthesis 103:118 the biosynthesis of polylactosamine-type N-glycans 103:152 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 7 50 theme SW620/5-FU 1007:1016 arg1 cells 1018:1022 SW620/5-FU cells 1007:1022 SW620/5-FU cells 1007:1022 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 11 51 contain has 1582:1584 arg2 implication 1598:1608 significant implication 1586:1608 significant implication for the development of new treatment strategies 1586:1656 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 11 51 contain has 1582:1584 arg1 alteration 1481:1490 the alteration 1477:1490 the alteration of β3GnT8 in CRC cells 1477:1513 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 5 52 theme inhibition 681:690 arg1 IC50 707:710 IC50 707:710 IC50 707:710 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 5 52 theme inhibition 681:690 arg1 concentration 692:704 50% inhibition concentration 677:704 50% inhibition concentration (IC50) of 5-FU 677:719 In this study, a 5-FU‑resistant cell line (SW620/5-FU) was generated, and 50% inhibition concentration (IC50) of 5-FU was determined by MTT assay. 25269761 10 53 theme 5-FU 1371:1374 arg1 resistance 1376:1385 5-FU resistance 1371:1385 5-FU resistance 1371:1385 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 11 54 theme strategies 1647:1656 arg1 development 1618:1628 the development 1614:1628 the development of new treatment strategies 1614:1656 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 1 55 theme Aberrant 155:162 arg1 glycosylation 164:176 Aberrant glycosylation 155:176 Aberrant glycosylation 155:176 Aberrant glycosylation is known to be associated with cancer chemoresistance. 25269761 6 56 theme blot 776:779 arg1 analysis 781:788 lectin blot analysis 769:788 lectin blot analysis 769:788 Flow cytometry and lectin blot analysis were performed to detect the alteration of polylactosamine structures. 25269761 10 57 theme RNA 1345:1347 arg1 interference 1349:1360 RNA interference 1345:1360 RNA interference 1345:1360 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25269761 0 58 theme cancer 75:80 arg1 cells 82:86 human colorectal cancer cells 58:86 human colorectal cancer cells 58:86 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 7 59 theme polylactosamine 988:1002 arg1 synthesis 975:983 the synthesis 971:983 the synthesis of polylactosamine in SW620/5-FU cells 971:1022 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 7 60 from synthesis 975:983 arg1 cells 1018:1022 SW620/5-FU cells 1007:1022 SW620/5-FU cells 1007:1022 Quantitative RT-‑PCR and western blot analysis were used to identify and evaluate candidate genes involved in the synthesis of polylactosamine in SW620/5-FU cells. 25269761 0 61 theme colorectal 64:73 arg1 cells 82:86 human colorectal cancer cells 58:86 human colorectal cancer cells 58:86 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 0 62 theme β3GnT8 13:18 arg1 Knockdown 0:8 Knockdown 0:8 Knockdown of β3GnT8 0:18 Knockdown of β3GnT8 reverses 5-fluorouracil resistance in human colorectal cancer cells via inhibition the biosynthesis of polylactosamine-type N-glycans. 25269761 11 63 theme new 1633:1635 arg1 strategies 1647:1656 new treatment strategies 1633:1656 new treatment strategies 1633:1656 In conclusion, the alteration of β3GnT8 in CRC cells correlates with tumor sensitivity to the chemotherapeutic drug and has significant implication for the development of new treatment strategies. 25269761 10 64 theme 5-FU‑resistant 1289:1302 arg1 cells 1311:1315 5-FU‑resistant cancer cells 1289:1315 5-FU‑resistant cancer cells 1289:1315 Further studies showed that β3GnT8 expression was also upregulated in 5-FU‑resistant cancer cells, and knockdown of β3GnT8 by RNA interference reversed 5-FU resistance through, at least partly, by suppressing the formation of polylactosamine. 25753708 2 0 theme N-glycosylation 313:327 arg1 model 304:308 the mathematical model 287:308 the mathematical model of N-glycosylation previously described by Krambeck et al. 287:367 It implements the mathematical model of N-glycosylation previously described by Krambeck et al. (Glycobiology 19:1163-1175, 2009). 25753708 0 1 theme GPP 72:74 arg1 Tool 77:80 the Glycan Pathway Predictor (GPP) Tool 42:80 the Glycan Pathway Predictor (GPP) Tool 42:80 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 6 2 theme pathway 1043:1049 arg1 set 999:1001 the set 995:1001 the set of potentially synthesized glycans 995:1036 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 6 2 theme pathway 1043:1049 arg1 glycans 1030:1036 potentially synthesized glycans 1006:1036 potentially synthesized glycans 1006:1036 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 6 2 theme pathway 1043:1049 arg1 map 1051:1053 a pathway map 1041:1053 a pathway map 1041:1053 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 1 3 theme Glycomes 212:219 arg1 INformatics 197:207 INformatics 197:207 INformatics of Glycomes 197:219 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 7 4 theme glyco-gene 1160:1169 arg1 experiment 1180:1189 a glyco-gene knockout experiment 1158:1189 a glyco-gene knockout experiment 1158:1189 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 3 5 theme glyco-enzymes 490:502 arg1 glyco-enzymes 490:502 glyco-enzymes 490:502 glyco-enzymes 490:502 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 3 5 theme glyco-enzymes 490:502 arg1 set 483:485 a select set 474:485 a select set of glyco-enzymes 474:502 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 6 6 theme glycan 914:919 arg1 structure 921:929 a particular starting glycan structure 892:929 a particular starting glycan structure as input 892:938 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 5 7 theme Detailed 680:687 arg1 results 689:695 Detailed results 680:695 Detailed results 680:695 Detailed results can also be obtained for each glycan that is theoretically generated, along with the corresponding enzymes that were found to be involved with the selected glycan. 25753708 7 8 theme spectrometric 1213:1225 arg1 data 1227:1230 mass spectrometric data 1208:1230 mass spectrometric data 1208:1230 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 3 9 theme select 476:481 arg1 glyco-enzymes 490:502 glyco-enzymes 490:502 glyco-enzymes 490:502 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 3 9 theme select 476:481 arg1 set 483:485 a select set 474:485 a select set of glyco-enzymes 474:502 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 7 10 theme tool 1095:1098 arg1 Applications 1074:1085 Applications 1074:1085 Applications of this tool 1074:1098 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 1 11 from http 241:244 arg1 website 230:236 website 230:236 website 230:236 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 0 12 theme glycan 10:15 arg1 synthesis 27:35 glycan structure synthesis 10:35 glycan structure synthesis 10:35 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 6 13 theme text 1063:1066 arg1 file 1068:1071 a text file 1061:1071 a text file 1061:1071 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 7 14 theme knockout 1171:1178 arg1 experiment 1180:1189 a glyco-gene knockout experiment 1158:1189 a glyco-gene knockout experiment 1158:1189 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 6 15 theme starting 905:912 arg1 structure 921:929 a particular starting glycan structure 892:929 a particular starting glycan structure as input 892:938 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 2 16 dep implements 276:285 arg1 Glycobiology 370:381 Glycobiology 370:381 Glycobiology 370:381 It implements the mathematical model of N-glycosylation previously described by Krambeck et al. (Glycobiology 19:1163-1175, 2009). 25753708 4 17 theme pathway 618:624 arg1 glycans 591:597 The resulting glycans 577:597 The resulting glycans 577:597 The resulting glycans are displayed as a pathway map, which can be evaluated in an interactive manner. 25753708 4 17 theme pathway 618:624 arg1 map 626:628 a pathway map 616:628 a pathway map 616:628 The resulting glycans are displayed as a pathway map, which can be evaluated in an interactive manner. 25753708 1 18 theme website 230:236 arg1 RINGS 177:181 the RINGS 173:181 the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/ 173:270 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 7 19 with comparison 1192:1201 arg1 data 1227:1230 mass spectrometric data 1208:1230 mass spectrometric data 1208:1230 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 7 20 theme mass 1208:1211 arg1 data 1227:1230 mass spectrometric data 1208:1230 mass spectrometric data 1208:1230 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 0 21 theme structure 17:25 arg1 synthesis 27:35 glycan structure synthesis 10:35 glycan structure synthesis 10:35 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 1 22 from Soka 224:227 arg1 Resource 184:191 Resource 184:191 Resource for INformatics of Glycomes at Soka 184:227 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 7 23 theme profiles 1145:1152 arg1 etc 1233:1235 etc 1233:1235 etc 1233:1235 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 7 23 theme profiles 1145:1152 arg1 comparison 1192:1201 comparison 1192:1201 comparison with mass spectrometric data 1192:1230 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 7 23 theme profiles 1145:1152 arg1 prediction 1124:1133 prediction 1124:1133 prediction of glycan profiles for a glyco-gene knockout experiment 1124:1189 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 25753708 4 24 theme resulting 581:589 arg1 glycans 591:597 The resulting glycans 577:597 The resulting glycans 577:597 The resulting glycans are displayed as a pathway map, which can be evaluated in an interactive manner. 25753708 4 24 theme resulting 581:589 arg1 map 626:628 a pathway map 616:628 a pathway map 616:628 The resulting glycans are displayed as a pathway map, which can be evaluated in an interactive manner. 25753708 5 25 theme selected 844:851 arg1 glycan 853:858 the selected glycan 840:858 the selected glycan 840:858 Detailed results can also be obtained for each glycan that is theoretically generated, along with the corresponding enzymes that were found to be involved with the selected glycan. 25753708 6 26 theme glyco-enzymes 957:969 arg1 glyco-enzymes 957:969 glyco-enzymes 957:969 glyco-enzymes 957:969 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 6 26 theme glyco-enzymes 957:969 arg1 set 950:952 a set 948:952 a set of glyco-enzymes 948:969 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 1 27 theme RINGS 177:181 arg1 part 165:168 a part 163:168 a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/ 163:270 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 0 28 theme Pathway 53:59 arg1 Tool 77:80 the Glycan Pathway Predictor (GPP) Tool 42:80 the Glycan Pathway Predictor (GPP) Tool 42:80 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 6 29 theme particular 894:903 arg1 structure 921:929 a particular starting glycan structure 892:929 a particular starting glycan structure as input 892:938 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 3 30 theme mathematical 516:527 arg1 model 529:533 a mathematical model 514:533 a mathematical model that characterizes substrate specificity 514:574 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 6 31 theme glycans 1030:1036 arg1 map 1051:1053 a pathway map 1041:1053 a pathway map 1041:1053 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 6 31 theme glycans 1030:1036 arg1 glycans 1030:1036 potentially synthesized glycans 1006:1036 potentially synthesized glycans 1006:1036 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 6 31 theme glycans 1030:1036 arg1 set 999:1001 the set 995:1001 the set of potentially synthesized glycans 995:1036 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 0 32 theme Glycan 46:51 arg1 Tool 77:80 the Glycan Pathway Predictor (GPP) Tool 42:80 the Glycan Pathway Predictor (GPP) Tool 42:80 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 2 33 theme mathematical 291:302 arg1 model 304:308 the mathematical model 287:308 the mathematical model of N-glycosylation previously described by Krambeck et al. 287:367 It implements the mathematical model of N-glycosylation previously described by Krambeck et al. (Glycobiology 19:1163-1175, 2009). 25753708 2 34 dep Krambeck 353:360 arg1 al 365:366 Krambeck et al 353:366 Krambeck et al 353:366 It implements the mathematical model of N-glycosylation previously described by Krambeck et al. (Glycobiology 19:1163-1175, 2009). 25753708 1 35 theme Glycan 110:115 arg1 Tool 135:138 the Glycan Pathway Predictor Tool 106:138 the Glycan Pathway Predictor Tool 106:138 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 1 35 theme Glycan 110:115 arg1 available 150:158 available 150:158 available 150:158 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 1 36 dep RINGS 177:181 arg1 Resource 184:191 Resource 184:191 Resource for INformatics of Glycomes at Soka 184:227 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 1 37 theme Pathway 117:123 arg1 Tool 135:138 the Glycan Pathway Predictor Tool 106:138 the Glycan Pathway Predictor Tool 106:138 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 1 37 theme Pathway 117:123 arg1 available 150:158 available 150:158 available 150:158 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 4 38 theme interactive 660:670 arg1 manner 672:677 an interactive manner 657:677 an interactive manner 657:677 The resulting glycans are displayed as a pathway map, which can be evaluated in an interactive manner. 25753708 5 39 theme corresponding 782:794 arg1 enzymes 796:802 the corresponding enzymes 778:802 the corresponding enzymes that were found to be involved with the selected glycan 778:858 Detailed results can also be obtained for each glycan that is theoretically generated, along with the corresponding enzymes that were found to be involved with the selected glycan. 25753708 3 40 theme substrate 554:562 arg1 specificity 564:574 substrate specificity 554:574 substrate specificity 554:574 This tool computes the glycans that can be potentially generated from a select set of glyco-enzymes, based on a mathematical model that characterizes substrate specificity. 25753708 1 41 theme Predictor 125:133 arg1 Tool 135:138 the Glycan Pathway Predictor Tool 106:138 the Glycan Pathway Predictor Tool 106:138 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 1 41 theme Predictor 125:133 arg1 available 150:158 available 150:158 available 150:158 This chapter describes the Glycan Pathway Predictor Tool, which is available as a part of the RINGS (Resource for INformatics of Glycomes at Soka) website at http://www.rings.t.soka.ac.jp/. 25753708 0 42 theme Predictor 61:69 arg1 Tool 77:80 the Glycan Pathway Predictor (GPP) Tool 42:80 the Glycan Pathway Predictor (GPP) Tool 42:80 Analyzing glycan structure synthesis with the Glycan Pathway Predictor (GPP) Tool. 25753708 6 43 theme synthesized 1018:1028 arg1 glycans 1030:1036 potentially synthesized glycans 1006:1036 potentially synthesized glycans 1006:1036 Thus, glycobiologists can take a particular starting glycan structure as input, select a set of glyco-enzymes, and instantly retrieve the set of potentially synthesized glycans as a pathway map or as a text file. 25753708 7 44 theme glycan 1138:1143 arg1 profiles 1145:1152 glycan profiles 1138:1152 glycan profiles for a glyco-gene knockout experiment 1138:1189 Applications of this tool are numerous, including prediction of glycan profiles for a glyco-gene knockout experiment, comparison with mass spectrometric data, etc. 28631470 5 0 theme 6-O-acetyl 859:868 arg1 mannopyranoside 870:884 p-nitrophenyl 6-O-acetyl mannopyranoside 845:884 p-nitrophenyl 6-O-acetyl mannopyranoside 845:884 The best yield was obtained for the furanosylation of p-nitrophenyl 6-O-acetyl mannopyranoside. 28631470 7 1 theme glycoconjugates 1119:1133 arg1 synthesis 1084:1092 the synthesis 1080:1092 the synthesis of biologically relevant glycoconjugates 1080:1133 This is a good alternative for the synthesis of biologically relevant glycoconjugates. 28631470 1 2 theme hydroxyl 181:188 arg1 group 190:194 more than one free hydroxyl group 162:194 more than one free hydroxyl group 162:194 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 0 3 theme Pathogenic 89:98 arg1 Microorganisms 100:113 Pathogenic Microorganisms 89:113 Pathogenic Microorganisms 89:113 Regioselective Galactofuranosylation for the Synthesis of Disaccharide Patterns Found in Pathogenic Microorganisms. 28631470 3 4 theme byproducts 577:586 arg1 formation 553:561 formation 553:561 formation of orthoester byproducts 553:586 Nevertheless formation of orthoester byproducts hampered the efficiency of the method. 28631470 2 5 from glycosylation 374:386 arg1 group 415:419 the equatorial hydroxyl group 391:419 the equatorial hydroxyl group of the acceptor 391:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 5 from glycosylation 374:386 arg1 acceptor 428:435 the acceptor 424:435 the acceptor 424:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 6 6 located found 922:926 arg1 glycocalix 935:944 the glycocalix 931:944 the glycocalix of some pathogens 931:962 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 6 6 located found 922:926 arg2 Precursors 887:896 Precursors 887:896 Precursors 887:896 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 4 7 contain containing 766:775 arg1 galactofuranosyl 749:764 the desired galactofuranosyl 737:764 the desired galactofuranosyl containing disaccharide 737:788 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 4 7 contain containing 766:775 arg2 disaccharide 777:788 disaccharide 777:788 disaccharide 777:788 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 2 8 theme High 344:347 arg1 regioselectivity 349:364 High regioselectivity 344:364 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor 344:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 7 9 theme good 1059:1062 arg1 alternative 1064:1074 a good alternative 1057:1074 a good alternative for the synthesis of biologically relevant glycoconjugates 1057:1133 This is a good alternative for the synthesis of biologically relevant glycoconjugates. 28631470 7 9 theme good 1059:1062 arg1 This 1049:1052 This 1049:1052 This 1049:1052 This is a good alternative for the synthesis of biologically relevant glycoconjugates. 28631470 6 10 theme same 999:1002 arg1 protocol 1004:1011 the same protocol 995:1011 the same protocol with yields ranging from 45 to 86% 995:1046 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 3 11 theme method 619:624 arg1 efficiency 601:610 the efficiency 597:610 the efficiency of the method 597:624 Nevertheless formation of orthoester byproducts hampered the efficiency of the method. 28631470 3 12 theme orthoester 566:575 arg1 byproducts 577:586 orthoester byproducts 566:586 orthoester byproducts 566:586 Nevertheless formation of orthoester byproducts hampered the efficiency of the method. 28631470 1 13 theme acceptors 147:155 arg1 glycosylation 130:142 Koenigs-Knorr glycosylation 116:142 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 116:243 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 6 14 theme disaccharides 907:919 arg1 Precursors 887:896 Precursors 887:896 Precursors 887:896 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 4 15 theme desired 741:747 arg1 galactofuranosyl 749:764 the desired galactofuranosyl 737:764 the desired galactofuranosyl containing disaccharide 737:788 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 1 16 gly glycosylation 130:142 arg1 acceptors 147:155 acceptors 147:155 acceptors with more than one free hydroxyl group 147:194 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 6 17 theme pathogens 954:962 arg1 glycocalix 935:944 the glycocalix 931:944 the glycocalix of some pathogens 931:962 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 0 18 theme Regioselective 0:13 arg1 Galactofuranosylation 15:35 Regioselective Galactofuranosylation 0:35 Regioselective Galactofuranosylation for the Synthesis of Disaccharide Patterns 0:78 Regioselective Galactofuranosylation for the Synthesis of Disaccharide Patterns Found in Pathogenic Microorganisms. 28631470 1 19 theme 2,3,5,6-tetrabenzoyl 199:218 arg1 bromide 237:243 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 199:243 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 199:243 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 4 20 from groups 662:667 arg1 O-6 672:674 O-6 672:674 O-6 672:674 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 4 20 from groups 662:667 arg1 C-1 682:684 C-1 682:684 C-1 of the acceptor 682:700 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 5 21 theme mannopyranoside 870:884 arg1 furanosylation 827:840 the furanosylation 823:840 the furanosylation of p-nitrophenyl 6-O-acetyl mannopyranoside 823:884 The best yield was obtained for the furanosylation of p-nitrophenyl 6-O-acetyl mannopyranoside. 28631470 1 22 with acceptors 147:155 arg1 group 190:194 more than one free hydroxyl group 162:194 more than one free hydroxyl group 162:194 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 1 23 theme galactofuranosyl 220:235 arg1 bromide 237:243 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 199:243 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 199:243 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 2 24 theme adduct 498:503 arg1 formation 474:482 the transient formation 460:482 the transient formation of a borinate adduct of the corresponding 1,2-cis diol 460:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 1 25 theme Koenigs-Knorr 116:128 arg1 glycosylation 130:142 Koenigs-Knorr glycosylation 116:142 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide 116:243 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 2 26 theme borinate 489:496 arg1 adduct 498:503 a borinate adduct 487:503 a borinate adduct of the corresponding 1,2-cis diol 487:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 1 27 theme regioselectivity 326:341 arg1 inducer 315:321 inducer 315:321 inducer of regioselectivity 315:341 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 5 28 theme best 795:798 arg1 yield 800:804 The best yield 791:804 The best yield 791:804 The best yield was obtained for the furanosylation of p-nitrophenyl 6-O-acetyl mannopyranoside. 28631470 2 29 theme corresponding 512:524 arg1 diol 534:537 the corresponding 1,2-cis diol 508:537 the corresponding 1,2-cis diol 508:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 30 theme diol 534:537 arg1 adduct 498:503 a borinate adduct 487:503 a borinate adduct of the corresponding 1,2-cis diol 487:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 4 31 theme electron-withdrawing 641:660 arg1 groups 662:667 electron-withdrawing groups 641:667 electron-withdrawing groups on O-6 or on C-1 of the acceptor 641:700 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 2 32 theme transient 464:472 arg1 formation 474:482 the transient formation 460:482 the transient formation of a borinate adduct of the corresponding 1,2-cis diol 460:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 33 theme 1,2-cis 526:532 arg1 diol 534:537 the corresponding 1,2-cis diol 508:537 the corresponding 1,2-cis diol 508:537 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 6 34 dep 86 1044:1045 arg1 to 1041:1042 to 1041:1042 to 1041:1042 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 1 35 theme diphenylborinic 265:279 arg1 ester 299:303 diphenylborinic acid 2-aminoethyl ester 265:303 diphenylborinic acid 2-aminoethyl ester (DPBA) 265:310 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 1 35 theme diphenylborinic 265:279 arg1 DPBA 306:309 DPBA 306:309 DPBA 306:309 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 0 36 theme Patterns 71:78 arg1 Synthesis 45:53 the Synthesis 41:53 the Synthesis of Disaccharide Patterns 41:78 Regioselective Galactofuranosylation for the Synthesis of Disaccharide Patterns Found in Pathogenic Microorganisms. 28631470 5 37 theme p-nitrophenyl 845:857 arg1 mannopyranoside 870:884 p-nitrophenyl 6-O-acetyl mannopyranoside 845:884 p-nitrophenyl 6-O-acetyl mannopyranoside 845:884 The best yield was obtained for the furanosylation of p-nitrophenyl 6-O-acetyl mannopyranoside. 28631470 2 38 theme hydroxyl 406:413 arg1 group 415:419 the equatorial hydroxyl group 391:419 the equatorial hydroxyl group of the acceptor 391:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 38 theme hydroxyl 406:413 arg1 acceptor 428:435 the acceptor 424:435 the acceptor 424:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 0 39 theme Disaccharide 58:69 arg1 Patterns 71:78 Disaccharide Patterns 58:78 Disaccharide Patterns 58:78 Regioselective Galactofuranosylation for the Synthesis of Disaccharide Patterns Found in Pathogenic Microorganisms. 28631470 2 40 theme equatorial 395:404 arg1 group 415:419 the equatorial hydroxyl group 391:419 the equatorial hydroxyl group of the acceptor 391:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 40 theme equatorial 395:404 arg1 acceptor 428:435 the acceptor 424:435 the acceptor 424:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 7 41 theme relevant 1110:1117 arg1 glycoconjugates 1119:1133 biologically relevant glycoconjugates 1097:1133 biologically relevant glycoconjugates 1097:1133 This is a good alternative for the synthesis of biologically relevant glycoconjugates. 28631470 6 42 theme other 901:905 arg1 disaccharides 907:919 other disaccharides 901:919 other disaccharides 901:919 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 1 43 theme acid 281:284 arg1 ester 299:303 diphenylborinic acid 2-aminoethyl ester 265:303 diphenylborinic acid 2-aminoethyl ester (DPBA) 265:310 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 1 43 theme acid 281:284 arg1 DPBA 306:309 DPBA 306:309 DPBA 306:309 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 6 44 with protocol 1004:1011 arg1 yields 1018:1023 yields 1018:1023 yields ranging from 45 to 86% 1018:1046 Precursors of other disaccharides, found in the glycocalix of some pathogens, were synthesized according to the same protocol with yields ranging from 45 to 86%. 28631470 1 45 theme 2-aminoethyl 286:297 arg1 ester 299:303 diphenylborinic acid 2-aminoethyl ester 265:303 diphenylborinic acid 2-aminoethyl ester (DPBA) 265:310 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 1 45 theme 2-aminoethyl 286:297 arg1 DPBA 306:309 DPBA 306:309 DPBA 306:309 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 28631470 2 46 theme acceptor 428:435 arg1 group 415:419 the equatorial hydroxyl group 391:419 the equatorial hydroxyl group of the acceptor 391:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 46 theme acceptor 428:435 arg1 acceptor 428:435 the acceptor 424:435 the acceptor 424:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 4 47 theme acceptor 693:700 arg1 C-1 682:684 C-1 682:684 C-1 of the acceptor 682:700 Interestingly electron-withdrawing groups on O-6 or on C-1 of the acceptor displaced the reaction in favor of the desired galactofuranosyl containing disaccharide. 28631470 2 48 gly glycosylation 374:386 arg1 group 415:419 the equatorial hydroxyl group 391:419 the equatorial hydroxyl group of the acceptor 391:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 2 48 gly glycosylation 374:386 arg1 acceptor 428:435 the acceptor 424:435 the acceptor 424:435 High regioselectivity for the glycosylation on the equatorial hydroxyl group of the acceptor was obtained thanks to the transient formation of a borinate adduct of the corresponding 1,2-cis diol. 28631470 1 49 theme free 176:179 arg1 group 190:194 more than one free hydroxyl group 162:194 more than one free hydroxyl group 162:194 Koenigs-Knorr glycosylation of acceptors with more than one free hydroxyl group by 2,3,5,6-tetrabenzoyl galactofuranosyl bromide was performed using diphenylborinic acid 2-aminoethyl ester (DPBA) as inducer of regioselectivity. 27960098 1 0 theme group 160:164 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 1 1 theme type 166:169 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 2 2 theme acceptor 376:383 arg1 elongation 334:343 the stepwise enzymatic chain elongation 305:343 the stepwise enzymatic chain elongation of readily available lactoside acceptor 305:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 2 3 theme stepwise 309:316 arg1 elongation 334:343 the stepwise enzymatic chain elongation 305:343 the stepwise enzymatic chain elongation of readily available lactoside acceptor 305:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 3 4 theme P1 611:612 arg1 epitopes 628:635 P1 trisaccharide epitopes 611:635 P1 trisaccharide epitopes 611:635 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 3 5 theme access 548:553 arg1 pentasaccharide 558:572 access P1 pentasaccharide 548:572 access P1 pentasaccharide 548:572 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 3 6 theme simple 529:534 arg1 approach 536:543 an operationally simple approach 512:543 an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes 512:635 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 3 7 theme related 595:601 arg1 Gb3 603:605 its structurally related Gb3 578:605 its structurally related Gb3 578:605 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 2 8 theme available 413:421 arg1 galactose 423:431 cheap and commercially available galactose 390:431 cheap and commercially available galactose 390:431 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 1 9 theme P1 171:172 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 1 10 theme enzymatic 73:81 arg1 synthesis 83:91 The enzymatic synthesis 69:91 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 69:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 1 11 theme pentasaccharide 174:188 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 1 12 theme antigen 190:196 arg1 synthesis 83:91 The enzymatic synthesis 69:91 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 69:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 0 13 theme Enzymatic 0:8 arg1 synthesis 10:18 Enzymatic synthesis 0:18 Enzymatic synthesis of human blood group P1 pentasaccharide antigen 0:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 14 theme chain 328:332 arg1 elongation 334:343 the stepwise enzymatic chain elongation 305:343 the stepwise enzymatic chain elongation of readily available lactoside acceptor 305:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 0 15 theme human 23:27 arg1 antigen 60:66 human blood group P1 pentasaccharide antigen 23:66 human blood group P1 pentasaccharide antigen 23:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 16 theme cheap 390:394 arg1 galactose 423:431 cheap and commercially available galactose 390:431 cheap and commercially available galactose 390:431 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 2 17 theme enzymatic 318:326 arg1 elongation 334:343 the stepwise enzymatic chain elongation 305:343 the stepwise enzymatic chain elongation of readily available lactoside acceptor 305:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 1 18 theme important 109:117 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 2 19 theme donor 460:464 arg1 precursors 466:475 donor precursors 460:475 donor precursors 460:475 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 0 20 theme group 35:39 arg1 antigen 60:66 human blood group P1 pentasaccharide antigen 23:66 human blood group P1 pentasaccharide antigen 23:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 3 21 theme P1 555:556 arg1 pentasaccharide 558:572 access P1 pentasaccharide 548:572 access P1 pentasaccharide 548:572 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 2 22 theme sequential 249:258 arg1 glycosylation 287:299 a three-step sequential one-pot multienzyme (OPME) glycosylation 236:299 a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor 236:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 0 23 theme blood 29:33 arg1 antigen 60:66 human blood group P1 pentasaccharide antigen 23:66 human blood group P1 pentasaccharide antigen 23:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 24 theme three-step 238:247 arg1 glycosylation 287:299 a three-step sequential one-pot multienzyme (OPME) glycosylation 236:299 a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor 236:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 0 25 theme pentasaccharide 44:58 arg1 antigen 60:66 human blood group P1 pentasaccharide antigen 23:66 human blood group P1 pentasaccharide antigen 23:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 26 with features 227:234 arg1 galactose 423:431 cheap and commercially available galactose 390:431 cheap and commercially available galactose 390:431 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 2 26 with features 227:234 arg1 N-acetylglucosamine 437:455 N-acetylglucosamine 437:455 N-acetylglucosamine 437:455 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 2 27 theme lactoside 366:374 arg1 acceptor 376:383 readily available lactoside acceptor 348:383 readily available lactoside acceptor 348:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 0 28 theme P1 41:42 arg1 antigen 60:66 human blood group P1 pentasaccharide antigen 23:66 human blood group P1 pentasaccharide antigen 23:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 29 theme available 356:364 arg1 acceptor 376:383 readily available lactoside acceptor 348:383 readily available lactoside acceptor 348:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 3 30 theme enzymatic 483:491 arg1 synthesis 493:501 This enzymatic synthesis 478:501 This enzymatic synthesis 478:501 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 2 31 theme OPME 281:284 arg1 glycosylation 287:299 a three-step sequential one-pot multienzyme (OPME) glycosylation 236:299 a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor 236:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 1 32 theme unique 136:141 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 3 33 theme trisaccharide 614:626 arg1 epitopes 628:635 P1 trisaccharide epitopes 611:635 P1 trisaccharide epitopes 611:635 This enzymatic synthesis provides an operationally simple approach to access P1 pentasaccharide and its structurally related Gb3 and P1 trisaccharide epitopes. 27960098 1 34 theme human 143:147 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 2 35 theme multienzyme 268:278 arg1 glycosylation 287:299 a three-step sequential one-pot multienzyme (OPME) glycosylation 236:299 a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor 236:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 1 36 theme P1PK 149:152 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 27960098 0 37 theme antigen 60:66 arg1 synthesis 10:18 Enzymatic synthesis 0:18 Enzymatic synthesis of human blood group P1 pentasaccharide antigen 0:66 Enzymatic synthesis of human blood group P1 pentasaccharide antigen. 27960098 2 38 theme one-pot 260:266 arg1 glycosylation 287:299 a three-step sequential one-pot multienzyme (OPME) glycosylation 236:299 a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor 236:383 This synthesis features a three-step sequential one-pot multienzyme (OPME) glycosylation for the stepwise enzymatic chain elongation of readily available lactoside acceptor with cheap and commercially available galactose and N-acetylglucosamine as donor precursors. 27960098 1 39 theme blood 154:158 arg1 antigen 190:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen 96:196 The enzymatic synthesis of biologically important and structurally unique human P1PK blood group type P1 pentasaccharide antigen is described. 26565680 3 0 contain contain 530:536 arg1 glycopeptide 513:524 a glycopeptide 511:524 a glycopeptide 511:524 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 3 0 contain contain 530:536 arg2 sites 561:565 multiple glycosylation sites 538:565 multiple glycosylation sites 538:565 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 3 0 contain contain 530:536 arg2 glycopeptide 513:524 a glycopeptide 511:524 a glycopeptide 511:524 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 3 0 contain contain 530:536 arg1 addition 501:508 addition 501:508 addition 501:508 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 4 1 theme spectral 736:743 arg1 algorithm 755:763 a graph-based spectral alignment algorithm 722:763 a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra 722:868 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 1 theme spectral 736:743 arg1 GlycoMID 712:719 GlycoMID 712:719 GlycoMID 712:719 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 5 2 gly glycopeptides 1095:1107 arg2 glycopeptides 1095:1107 many glycopeptides 1090:1107 many glycopeptides with two glycosylation sites 1090:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 0 3 theme Mass 92:95 arg1 Spectrometry 97:108 Tandem Mass Spectrometry 85:108 Tandem Mass Spectrometry 85:108 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 3 4 gly glycopeptide 513:524 arg2 glycopeptide 513:524 a glycopeptide 511:524 a glycopeptide 511:524 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 3 5 theme multiple 538:545 arg1 sites 561:565 multiple glycosylation sites 538:565 multiple glycosylation sites 538:565 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 5 6 gly glycosylation 1118:1130 arg2 sites 1132:1136 two glycosylation sites 1114:1136 two glycosylation sites 1114:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 6 gly glycosylation 1118:1130 arg2 two 1114:1116 two 1114:1116 two 1114:1116 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 4 7 theme tandem 850:855 arg1 spectra 862:868 tandem mass spectra 850:868 tandem mass spectra 850:868 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 8 with glycopeptides 642:654 arg1 sites 684:688 multiple glycosylation sites 661:688 multiple glycosylation sites 661:688 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 3 9 theme glycosylation 547:559 arg1 sites 561:565 multiple glycosylation sites 538:565 multiple glycosylation sites 538:565 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 2 10 theme efficient 308:316 arg1 spectrometry 289:300 mass spectrometry 284:300 mass spectrometry 284:300 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 10 theme efficient 308:316 arg1 technique 318:326 an efficient technique 305:326 an efficient technique for glycopeptide identification 305:358 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 4 11 gly glycopeptides 783:795 arg2 glycopeptides 783:795 glycopeptides 783:795 glycopeptides with multiple hydroxylysine O-glycosylation sites 783:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 12 theme glycosylation 670:682 arg1 sites 684:688 multiple glycosylation sites 661:688 multiple glycosylation sites 661:688 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 3 13 theme problem 579:585 arg1 complex 587:593 the problem complex 575:593 the problem complex 575:593 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 3 14 gly glycosylation 547:559 arg2 sites 561:565 multiple glycosylation sites 538:565 multiple glycosylation sites 538:565 In addition, a glycopeptide may contain multiple glycosylation sites, making the problem complex. 26565680 4 15 gly glycosylation 670:682 arg2 sites 684:688 multiple glycosylation sites 661:688 multiple glycosylation sites 661:688 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 1 16 from modifications 170:182 arg1 proteins 187:194 proteins 187:194 proteins 187:194 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 5 17 theme more 1018:1021 arg1 matches 1045:1051 more glycopeptide-spectrum matches 1018:1051 more glycopeptide-spectrum matches 1018:1051 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 2 18 theme enormous 419:426 arg1 diversity 428:436 the enormous diversity 415:436 the enormous diversity of glycan structures 415:457 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 0 19 theme O-Glycosylation 60:74 arg1 Sites 76:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 5 20 theme glycopeptide-spectrum 1023:1043 arg1 matches 1045:1051 more glycopeptide-spectrum matches 1018:1051 more glycopeptide-spectrum matches 1018:1051 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 21 with glycopeptides 1095:1107 arg1 sites 1132:1136 two glycosylation sites 1114:1136 two glycosylation sites 1114:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 0 22 theme Tandem 85:90 arg1 Spectrometry 97:108 Tandem Mass Spectrometry 85:108 Tandem Mass Spectrometry 85:108 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 5 23 theme experimental 971:982 arg1 results 984:990 experimental results 971:990 experimental results 971:990 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 24 theme protein 958:964 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 24 theme protein 958:964 arg1 sets 917:920 mass spectrometry data sets 894:920 mass spectrometry data sets of the bovine collagen α-(II) chain protein 894:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 25 theme bovine 929:934 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 26 theme spectrometry 899:910 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 26 theme spectrometry 899:910 arg1 sets 917:920 mass spectrometry data sets 894:920 mass spectrometry data sets of the bovine collagen α-(II) chain protein 894:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 2 27 theme glycopeptide 332:343 arg1 identification 345:358 glycopeptide identification 332:358 glycopeptide identification 332:358 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 5 28 theme collagen 936:943 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 4 29 theme multiple 802:809 arg1 sites 841:845 multiple hydroxylysine O-glycosylation sites 802:845 multiple hydroxylysine O-glycosylation sites 802:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 2 30 theme structures 448:457 arg1 microheterogeneity 467:484 the microheterogeneity 463:484 the microheterogeneity of glycans 463:495 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 30 theme structures 448:457 arg1 diversity 428:436 the enormous diversity 415:436 the enormous diversity of glycan structures 415:457 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 0 31 theme Glycopeptides 18:30 arg1 Identification 0:13 Identification 0:13 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 0:109 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 4 32 theme software 604:611 arg1 tools 613:617 Current software tools 596:617 Current software tools 596:617 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 33 theme graph-based 724:734 arg1 algorithm 755:763 a graph-based spectral alignment algorithm 722:763 a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra 722:868 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 33 theme graph-based 724:734 arg1 GlycoMID 712:719 GlycoMID 712:719 GlycoMID 712:719 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 2 34 theme glycan 441:446 arg1 structures 448:457 glycan structures 441:457 glycan structures 441:457 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 0 35 theme Hydroxylysine 46:58 arg1 Sites 76:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 4 36 theme Current 596:602 arg1 tools 613:617 Current software tools 596:617 Current software tools 596:617 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 5 37 theme data 912:915 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 37 theme data 912:915 arg1 sets 917:920 mass spectrometry data sets 894:920 mass spectrometry data sets of the bovine collagen α-(II) chain protein 894:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 1 38 theme mammalian 217:225 arg1 proteins 227:234 mammalian proteins 217:234 mammalian proteins 217:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 0 39 theme Multiple 37:44 arg1 Sites 76:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 4 40 theme hydroxylysine 811:823 arg1 sites 841:845 multiple hydroxylysine O-glycosylation sites 802:845 multiple hydroxylysine O-glycosylation sites 802:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 2 41 theme research 245:252 arg1 groups 254:259 Several research groups 237:259 Several research groups 237:259 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 1 42 theme proteins 227:234 arg1 % 212:212 ~50% 209:212 ~50% of mammalian proteins 209:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 1 42 theme proteins 227:234 arg1 proteins 227:234 mammalian proteins 217:234 mammalian proteins 217:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 5 43 theme α- 945:946 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 2 44 theme Several 237:243 arg1 groups 254:259 Several research groups 237:259 Several research groups 237:259 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 0 45 with Glycopeptides 18:30 arg1 Sites 76:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Multiple Hydroxylysine O-Glycosylation Sites 37:80 Identification of Glycopeptides with Multiple Hydroxylysine O-Glycosylation Sites by Tandem Mass Spectrometry. 26565680 5 46 theme chain 952:956 arg1 protein 958:964 the bovine collagen α-(II) chain protein 925:964 the bovine collagen α-(II) chain protein 925:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 2 47 theme glycans 489:495 arg1 microheterogeneity 467:484 the microheterogeneity 463:484 the microheterogeneity of glycans 463:495 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 47 theme glycans 489:495 arg1 diversity 428:436 the enormous diversity 415:436 the enormous diversity of glycan structures 415:457 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 4 48 with glycopeptides 783:795 arg1 sites 841:845 multiple hydroxylysine O-glycosylation sites 802:845 multiple hydroxylysine O-glycosylation sites 802:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 5 49 theme many 1090:1093 arg1 glycopeptides 1095:1107 many glycopeptides 1090:1107 many glycopeptides with two glycosylation sites 1090:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 4 50 theme alignment 745:753 arg1 algorithm 755:763 a graph-based spectral alignment algorithm 722:763 a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra 722:868 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 50 theme alignment 745:753 arg1 GlycoMID 712:719 GlycoMID 712:719 GlycoMID 712:719 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 2 51 gly microheterogeneity 467:484 arg1 glycans 489:495 glycans 489:495 glycans 489:495 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 51 gly microheterogeneity 467:484 arg1 structures 448:457 glycan structures 441:457 glycan structures 441:457 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 52 theme mass 284:287 arg1 spectrometry 289:300 mass spectrometry 284:300 mass spectrometry 284:300 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 2 52 theme mass 284:287 arg1 technique 318:326 an efficient technique 305:326 an efficient technique for glycopeptide identification 305:358 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 4 53 gly glycopeptides 642:654 arg2 glycopeptides 642:654 glycopeptides 642:654 glycopeptides with multiple glycosylation sites 642:688 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 54 theme O-glycosylation 825:839 arg1 sites 841:845 multiple hydroxylysine O-glycosylation sites 802:845 multiple hydroxylysine O-glycosylation sites 802:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 1 55 theme common 144:149 arg1 modifications 170:182 the most common post-translational modifications 135:182 the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins 135:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 4 56 theme mass 857:860 arg1 spectra 862:868 tandem mass spectra 850:868 tandem mass spectra 850:868 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 2 57 gly glycopeptide 332:343 arg2 glycopeptide 332:343 glycopeptide identification 332:358 glycopeptide identification 332:358 Several research groups have demonstrated that mass spectrometry is an efficient technique for glycopeptide identification; however, this problem is still challenging because of the enormous diversity of glycan structures and the microheterogeneity of glycans. 26565680 5 58 theme glycosylation 1118:1130 arg1 sites 1132:1136 two glycosylation sites 1114:1136 two glycosylation sites 1114:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 59 theme other 1058:1062 arg1 glycopeptides 1095:1107 many glycopeptides 1090:1107 many glycopeptides with two glycosylation sites 1090:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 59 theme other 1058:1062 arg1 tools 1073:1077 other existing tools 1058:1077 other existing tools 1058:1077 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 60 theme mass 894:897 arg1 spectrometry 899:910 mass spectrometry 894:910 mass spectrometry data sets of the bovine collagen α-(II) chain protein 894:964 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 1 61 theme post-translational 151:168 arg1 modifications 170:182 the most common post-translational modifications 135:182 the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins 135:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 4 62 theme multiple 661:668 arg1 sites 684:688 multiple glycosylation sites 661:688 multiple glycosylation sites 661:688 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 4 63 gly O-glycosylation 825:839 arg2 sites 841:845 multiple hydroxylysine O-glycosylation sites 802:845 multiple hydroxylysine O-glycosylation sites 802:845 Current software tools often fail to identify glycopeptides with multiple glycosylation sites, and hence we present GlycoMID, a graph-based spectral alignment algorithm that can identify glycopeptides with multiple hydroxylysine O-glycosylation sites by tandem mass spectra. 26565680 5 64 theme existing 1064:1071 arg1 glycopeptides 1095:1107 many glycopeptides 1090:1107 many glycopeptides with two glycosylation sites 1090:1136 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 5 64 theme existing 1064:1071 arg1 tools 1073:1077 other existing tools 1058:1077 other existing tools 1058:1077 GlycoMID was tested on mass spectrometry data sets of the bovine collagen α-(II) chain protein, and experimental results showed that it identified more glycopeptide-spectrum matches than other existing tools, including many glycopeptides with two glycosylation sites. 26565680 1 65 theme modifications 170:182 arg1 modifications 170:182 the most common post-translational modifications 135:182 the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins 135:234 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 26565680 1 65 theme modifications 170:182 arg1 one 128:130 one 128:130 one 128:130 Glycosylation is one of the most common post-translational modifications in proteins, existing in ~50% of mammalian proteins. 25688550 2 0 theme Und-PP-GalNAc2 491:504 arg1 Bac1 506:509 Und-PP-GalNAc2 Bac1 491:509 Und-PP-GalNAc2 Bac1 491:509 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 3 1 theme tandem 650:655 arg1 MS 657:658 tandem MS 650:658 tandem MS to confirm their chemical structures 650:695 The products were analyzed in detail by tandem MS to confirm their chemical structures. 25688550 2 2 theme tagged 418:423 arg1 peptides 425:432 fluorescently tagged peptides 404:432 fluorescently tagged peptides 404:432 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 0 3 theme Campylobacter 137:149 arg1 jejuni 151:156 Campylobacter jejuni 137:156 Campylobacter jejuni 137:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 1 4 from jejuni 213:218 arg1 PglB 189:192 The oligosaccharyltransferase PglB 159:192 The oligosaccharyltransferase PglB from Campylobacter jejuni 159:218 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 2 5 theme biosynthetic 449:460 arg1 Und-PP-Bac1 475:485 biosynthetic intermediate Und-PP-Bac1 449:485 biosynthetic intermediate Und-PP-Bac1 449:485 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 1 6 link undecaprenyl-pyrophosphate-linked 264:296 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 6 link undecaprenyl-pyrophosphate-linked 264:296 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 2 7 theme Asn 546:548 arg1 residue 550:556 the Asn residue 542:556 the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P) 542:607 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 2 8 dep sequence 575:582 arg1 X 601:601 X' 601:602 X' 601:602 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 2 8 dep sequence 575:582 arg1 X≠P 604:606 X≠P 604:606 X≠P 604:606 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 2 8 dep sequence 575:582 arg1 /S 597:598 D/E-X'-N-X-T/S 585:598 D/E-X'-N-X-T/S 585:598 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 1 9 theme undecaprenyl-pyrophosphate-linked 264:296 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 9 theme undecaprenyl-pyrophosphate-linked 264:296 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 2 10 theme D/E-X'-N-X-T 585:596 arg1 X 601:601 X' 601:602 X' 601:602 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 2 10 theme D/E-X'-N-X-T 585:596 arg1 X≠P 604:606 X≠P 604:606 X≠P 604:606 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 2 10 theme D/E-X'-N-X-T 585:596 arg1 /S 597:598 D/E-X'-N-X-T/S 585:598 D/E-X'-N-X-T/S 585:598 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 3 11 theme chemical 677:684 arg1 structures 686:695 their chemical structures 671:695 their chemical structures 671:695 The products were analyzed in detail by tandem MS to confirm their chemical structures. 25688550 1 12 theme Glc1 298:301 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 12 theme Glc1 298:301 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 0 13 theme undecaprenyl 31:42 arg1 oligosaccharide 65:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide 21:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni 21:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 0 14 link pyrophosphate-linked 44:63 arg1 oligosaccharide 65:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide 21:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni 21:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 1 15 theme GalNAc5 303:309 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 15 theme GalNAc5 303:309 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 0 16 theme synthetic 21:29 arg1 oligosaccharide 65:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide 21:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni 21:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 1 17 theme N-glycosylation 234:248 arg1 reaction 250:257 the N-glycosylation reaction 230:257 the N-glycosylation reaction 230:257 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 2 18 from residue 550:556 arg1 sequence 575:582 the consensus sequence 561:582 the consensus sequence (D/E-X'-N-X-T/S, X',X≠P) 561:607 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 0 19 theme pyrophosphate-linked 44:63 arg1 oligosaccharide 65:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide 21:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni 21:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 1 20 theme Und-PP-Glc1 317:327 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 20 theme Und-PP-Glc1 317:327 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 21 theme oligosaccharyltransferase 163:187 arg1 PglB 189:192 The oligosaccharyltransferase PglB 159:192 The oligosaccharyltransferase PglB from Campylobacter jejuni 159:218 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 2 22 theme synthesized 374:384 arg1 donors 386:391 chemically synthesized donors 363:391 chemically synthesized donors coupled to fluorescently tagged peptides 363:432 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 0 23 with N-Glycosylation 0:14 arg1 oligosaccharide 65:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide 21:79 synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni 21:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 2 24 theme intermediate 462:473 arg1 Und-PP-Bac1 475:485 biosynthetic intermediate Und-PP-Bac1 449:485 biosynthetic intermediate Und-PP-Bac1 449:485 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 25688550 1 25 theme GalNAc5 329:335 arg1 Bac1 311:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 264:314 undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ) 264:342 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 1 25 theme GalNAc5 329:335 arg1 Bac1 337:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 Und-PP-Glc1 GalNAc5 Bac1 317:340 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 0 26 from jejuni 151:156 arg1 oligosaccharyltransferase 106:130 PglB oligosaccharyltransferase 101:130 PglB oligosaccharyltransferase from Campylobacter jejuni 101:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 1 27 theme Campylobacter 199:211 arg1 jejuni 213:218 Campylobacter jejuni 199:218 Campylobacter jejuni 199:218 The oligosaccharyltransferase PglB from Campylobacter jejuni catalyses the N-glycosylation reaction with undecaprenyl-pyrophosphate-linked Glc1 GalNAc5 Bac1 (Und-PP-Glc1 GalNAc5 Bac1 ). 25688550 0 28 theme PglB 101:104 arg1 oligosaccharyltransferase 106:130 PglB oligosaccharyltransferase 101:130 PglB oligosaccharyltransferase from Campylobacter jejuni 101:156 N-Glycosylation with synthetic undecaprenyl pyrophosphate-linked oligosaccharide to oligopeptides by PglB oligosaccharyltransferase from Campylobacter jejuni. 25688550 2 29 theme consensus 565:573 arg1 sequence 575:582 the consensus sequence 561:582 the consensus sequence (D/E-X'-N-X-T/S, X',X≠P) 561:607 Experiments using chemically synthesized donors coupled to fluorescently tagged peptides confirmed that biosynthetic intermediate Und-PP-Bac1 and Und-PP-GalNAc2 Bac1 are transferred efficiently to the Asn residue in the consensus sequence (D/E-X'-N-X-T/S, X',X≠P). 26831718 8 0 theme discriminative 1232:1245 arg1 AUC 1254:1256 AUC = 0.499 1254:1264 AUC = 0.499 1254:1264 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 8 0 theme discriminative 1232:1245 arg1 power 1247:1251 discriminative power 1232:1251 discriminative power (AUC = 0.499) 1232:1265 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 7 1 gly sialylation 1047:1057 arg1 core-fucosylation 1075:1091 core-fucosylation 1075:1091 core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1075:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 5 2 theme predictive 703:712 arg1 model 714:718 A predictive model 701:718 A predictive model 701:718 A predictive model was built using regularized logistic regression and evaluated using a 10-cross validation procedure. 26831718 3 3 theme colorectal 561:570 arg1 cancer 572:577 colorectal cancer 561:577 colorectal cancer 561:577 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 10 4 from variation 1671:1679 arg1 glycosylation 1688:1700 IgG glycosylation 1684:1700 IgG glycosylation 1684:1700 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 5 with immunosurveillance 1597:1614 arg1 mAbs 1654:1657 mAbs 1654:1657 mAbs 1654:1657 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 8 6 theme = 1258:1258 arg1 AUC 1254:1256 AUC = 0.499 1254:1264 AUC = 0.499 1254:1264 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 8 6 theme = 1258:1258 arg1 power 1247:1251 discriminative power 1232:1251 discriminative power (AUC = 0.499) 1232:1265 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 7 7 dep RESULTS 949:955 arg1 found 965:969 found 965:969 have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 960:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 3 8 theme matching 587:594 arg1 controls 596:603 538 matching controls 583:603 538 matching controls 583:603 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 3 9 with patients 547:554 arg1 cancer 572:577 colorectal cancer 561:577 colorectal cancer 561:577 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 1 10 contain has 68:70 arg1 glycosylation 54:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 1 10 contain has 68:70 arg2 consequences 110:121 significant structural and functional consequences 72:121 significant structural and functional consequences 72:121 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 10 11 theme detailed 1820:1827 arg1 studies 1829:1835 further, more detailed studies 1806:1835 studies 1829:1835 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 7 12 theme sialylated 1161:1170 arg1 glycans 1172:1178 sialylated glycans 1161:1178 sialylated glycans 1161:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 2 13 theme individual 252:261 arg1 variations 263:272 highly heritable individual variations 235:272 highly heritable individual variations 235:272 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 7 14 theme glycans 1104:1110 arg1 core-fucosylation 1075:1091 core-fucosylation 1075:1091 core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1075:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 8 15 theme variables 1291:1299 arg1 addition 1272:1279 the addition 1268:1279 the addition of glycan variables into the model 1268:1314 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 5 16 theme regularized 736:746 arg1 regression 757:766 regularized logistic regression 736:766 regularized logistic regression 736:766 A predictive model was built using regularized logistic regression and evaluated using a 10-cross validation procedure. 26831718 7 17 gly core-fucosylation 1140:1156 arg1 glycans 1172:1178 sialylated glycans 1161:1178 sialylated glycans 1161:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 10 18 theme IgG 1684:1686 arg1 glycosylation 1688:1700 IgG glycosylation 1684:1700 IgG glycosylation 1684:1700 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 7 19 theme core-fucosylation 1140:1156 arg1 decrease 1128:1135 concurrent decrease 1117:1135 concurrent decrease of core-fucosylation of sialylated glycans 1117:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 10 20 theme glycosylation 1844:1856 arg1 studies 1829:1835 further, more detailed studies 1806:1835 studies 1829:1835 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 2 21 from dynamics 301:308 arg1 glycosylation 328:340 IgG glycosylation 324:340 IgG glycosylation on colorectal cancer 324:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 5 22 theme logistic 748:755 arg1 regression 757:766 regularized logistic regression 736:766 regularized logistic regression 736:766 A predictive model was built using regularized logistic regression and evaluated using a 10-cross validation procedure. 26831718 2 23 from changes 313:319 arg1 glycosylation 328:340 IgG glycosylation 324:340 IgG glycosylation on colorectal cancer 324:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 2 23 from changes 313:319 arg1 cancer 356:361 colorectal cancer 345:361 colorectal cancer 345:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 10 24 theme disease 1749:1755 arg1 course 1757:1762 disease course 1749:1762 disease course 1749:1762 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 8 25 theme = 1382:1382 arg1 AUC 1378:1380 AUC = 0.755 1378:1388 AUC = 0.755 1378:1388 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 1 26 theme cancer 155:160 arg1 immunosurveillance 162:179 cancer immunosurveillance 155:179 cancer immunosurveillance 155:179 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 10 27 from studies 1829:1835 arg1 cancer 1872:1877 colorectal cancer 1861:1877 colorectal cancer 1861:1877 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 7 28 gly sialylated 1161:1170 arg1 glycans 1172:1178 sialylated glycans 1161:1178 sialylated glycans 1161:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 7 29 theme IgG 1043:1045 arg1 sialylation 1047:1057 IgG sialylation 1043:1057 IgG sialylation 1043:1057 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 5 30 theme 10-cross 790:797 arg1 procedure 810:818 a 10-cross validation procedure 788:818 a 10-cross validation procedure 788:818 A predictive model was built using regularized logistic regression and evaluated using a 10-cross validation procedure. 26831718 11 31 dep 22 1897:1898 arg1 3078-86 1905:1911 3078-86 1905:1911 3078-86 1905:1911 Clin Cancer Res; 22(12); 3078-86. 26831718 12 32 theme ©2016 1914:1918 arg1 AACR 1920:1923 ©2016 AACR 1914:1923 ©2016 AACR. 1914:1924 ©2016 AACR. 26831718 1 33 theme structural 84:93 arg1 consequences 110:121 significant structural and functional consequences 72:121 significant structural and functional consequences 72:121 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 3 34 theme IgG 476:478 arg1 analysis 494:501 IgG glycosylation analysis 476:501 IgG glycosylation analysis 476:501 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 1 35 theme functional 99:108 arg1 consequences 110:121 significant structural and functional consequences 72:121 significant structural and functional consequences 72:121 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 10 36 theme IgG 1564:1566 arg1 glycosylation 1568:1580 IgG glycosylation 1564:1580 IgG glycosylation 1564:1580 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 3 37 theme IgG 516:518 arg1 composition 528:538 IgG glycome composition 516:538 IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls 516:603 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 7 38 from galactosylation 1026:1040 arg1 core-fucosylation 1075:1091 core-fucosylation 1075:1091 core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1075:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 4 39 theme surgery 617:623 arg1 Effects 606:612 Effects 606:612 Effects of surgery 606:623 Effects of surgery were evaluated in 28 patients sampled before and three times after surgery. 26831718 2 40 theme technological 193:205 arg1 limitations 207:217 technological limitations 193:217 technological limitations 193:217 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 6 41 theme colorectal 930:939 arg1 cancer 941:946 colorectal cancer 930:946 colorectal cancer 930:946 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 3 42 theme high-throughput 440:454 arg1 technology 461:470 recently developed high-throughput UPLC technology 421:470 recently developed high-throughput UPLC technology 421:470 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 10 43 theme tumor 1591:1595 arg1 immunosurveillance 1597:1614 tumor immunosurveillance 1591:1614 tumor immunosurveillance 1591:1614 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 44 theme choice 1771:1776 arg1 prediction 1735:1744 prediction 1735:1744 prediction of disease course or the choice of therapy 1735:1787 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 9 45 theme differences 1415:1425 arg1 none 1401:1404 none 1401:1404 none of these differences 1401:1425 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 3 46 from composition 528:538 arg1 controls 596:603 538 matching controls 583:603 538 matching controls 583:603 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 3 46 from composition 528:538 arg1 patients 547:554 760 patients 543:554 760 patients with colorectal cancer 543:577 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 2 47 gly glycosylation 328:340 arg1 cancer 356:361 colorectal cancer 345:361 colorectal cancer 345:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 7 48 theme colorectal 976:985 arg1 cancer 987:992 colorectal cancer 976:992 colorectal cancer 976:992 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 1 49 theme Alternative 42:52 arg1 glycosylation 54:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 10 50 theme therapy 1781:1787 arg1 course 1757:1762 disease course 1749:1762 disease course 1749:1762 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 50 theme therapy 1781:1787 arg1 choice 1771:1776 the choice 1767:1776 the choice of therapy 1767:1787 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 2 51 from cancer 356:361 arg1 changes 313:319 changes 313:319 changes in IgG glycosylation on colorectal cancer 313:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 10 52 dep turn 1706:1709 arg1 out 1711:1713 out 1711:1713 out 1711:1713 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 6 53 theme plasma 877:882 arg1 samples 884:890 39 plasma samples 874:890 39 plasma samples collected before initial diagnosis of colorectal cancer 874:946 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 2 54 theme changes 313:319 arg1 dynamics 301:308 the dynamics 297:308 the dynamics of changes in IgG glycosylation on colorectal cancer 297:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 9 55 theme samples 1464:1470 arg1 samples 1464:1470 samples 1464:1470 samples collected before the initial diagnosis 1464:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 9 55 theme samples 1464:1470 arg1 set 1457:1459 the small set 1447:1459 the small set of samples collected before the initial diagnosis 1447:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 10 56 with efficacy 1629:1636 arg1 mAbs 1654:1657 mAbs 1654:1657 mAbs 1654:1657 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 57 theme therapy 1641:1647 arg1 immunosurveillance 1597:1614 tumor immunosurveillance 1591:1614 tumor immunosurveillance 1591:1614 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 57 theme therapy 1641:1647 arg1 efficacy 1629:1636 clinical efficacy 1620:1636 clinical efficacy 1620:1636 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 10 58 theme further 1806:1812 arg1 studies 1829:1835 further, more detailed studies 1806:1835 studies 1829:1835 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 2 59 from effects 224:230 arg1 dynamics 301:308 the dynamics 297:308 the dynamics of changes in IgG glycosylation on colorectal cancer 297:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 7 60 from increase 1063:1070 arg1 core-fucosylation 1075:1091 core-fucosylation 1075:1091 core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1075:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 10 61 theme IgG 1840:1842 arg1 glycosylation 1844:1856 IgG glycosylation 1840:1856 IgG glycosylation 1840:1856 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 9 62 theme initial 1493:1499 arg1 diagnosis 1501:1509 the initial diagnosis 1489:1509 the initial diagnosis 1489:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 10 63 theme individual 1660:1669 arg1 variation 1671:1679 individual variation 1660:1679 individual variation in IgG glycosylation 1660:1700 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 7 64 theme glycans 1172:1178 arg1 core-fucosylation 1140:1156 core-fucosylation 1140:1156 core-fucosylation of sialylated glycans 1140:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 6 65 theme glycome 838:844 arg1 composition 846:856 IgG glycome composition 834:856 IgG glycome composition 834:856 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 2 66 theme IgG 324:326 arg1 glycosylation 328:340 IgG glycosylation 324:340 IgG glycosylation on colorectal cancer 324:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 2 67 theme variations 263:272 arg1 differences 282:292 the differences 278:292 the differences in the dynamics of changes in IgG glycosylation on colorectal cancer 278:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 2 67 theme variations 263:272 arg1 effects 224:230 the effects 220:230 the effects of highly heritable individual variations 220:272 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 8 68 theme discriminative 1343:1356 arg1 power 1358:1362 the discriminative power 1339:1362 the discriminative power of the model 1339:1375 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 2 69 theme heritable 242:250 arg1 variations 263:272 highly heritable individual variations 235:272 highly heritable individual variations 235:272 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 8 70 theme glycan 1284:1289 arg1 variables 1291:1299 glycan variables 1284:1299 glycan variables 1284:1299 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 7 71 from decrease 1010:1017 arg1 increase 1063:1070 increase 1063:1070 increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1063:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 7 71 from decrease 1010:1017 arg1 galactosylation 1026:1040 IgG galactosylation 1022:1040 IgG galactosylation 1022:1040 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 7 71 from decrease 1010:1017 arg1 sialylation 1047:1057 IgG sialylation 1043:1057 IgG sialylation 1043:1057 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 7 72 theme concurrent 1117:1126 arg1 decrease 1128:1135 concurrent decrease 1117:1135 concurrent decrease of core-fucosylation of sialylated glycans 1117:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 7 73 theme neutral 1096:1102 arg1 glycans 1104:1110 neutral glycans 1096:1110 neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1096:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 8 74 theme model 1371:1375 arg1 power 1358:1362 the discriminative power 1339:1362 the discriminative power of the model 1339:1375 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 10 75 theme glycosylation 1568:1580 arg1 relevance 1551:1559 the functional relevance 1536:1559 the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs 1536:1657 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 0 76 from Glycome 4:10 arg1 Cancer 26:31 Colorectal Cancer 15:31 Colorectal Cancer 15:31 IgG Glycome in Colorectal Cancer. 26831718 7 77 from sialylation 1047:1057 arg1 core-fucosylation 1075:1091 core-fucosylation 1075:1091 core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1075:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 2 78 from glycosylation 328:340 arg1 dynamics 301:308 the dynamics 297:308 the dynamics of changes in IgG glycosylation on colorectal cancer 297:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 2 78 from glycosylation 328:340 arg1 cancer 356:361 colorectal cancer 345:361 colorectal cancer 345:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 7 79 with glycans 1104:1110 arg1 decrease 1128:1135 concurrent decrease 1117:1135 concurrent decrease of core-fucosylation of sialylated glycans 1117:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 3 80 theme EXPERIMENTAL 395:406 arg1 DESIGN 408:413 EXPERIMENTAL DESIGN 395:413 EXPERIMENTAL DESIGN 395:413 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 10 81 theme course 1757:1762 arg1 prediction 1735:1744 prediction 1735:1744 prediction of disease course or the choice of therapy 1735:1787 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 5 82 theme validation 799:808 arg1 procedure 810:818 a 10-cross validation procedure 788:818 a 10-cross validation procedure 788:818 A predictive model was built using regularized logistic regression and evaluated using a 10-cross validation procedure. 26831718 1 83 theme significant 72:82 arg1 consequences 110:121 significant structural and functional consequences 72:121 significant structural and functional consequences 72:121 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 11 84 dep Res 1892:1894 arg1 12 1900:1901 12 1900:1901 12 1900:1901 Clin Cancer Res; 22(12); 3078-86. 26831718 11 84 dep Res 1892:1894 arg1 22 1897:1898 22 1897:1898 22 1897:1898 Clin Cancer Res; 22(12); 3078-86. 26831718 7 85 theme IgG 1022:1024 arg1 galactosylation 1026:1040 IgG galactosylation 1022:1040 IgG galactosylation 1022:1040 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 3 86 theme developed 430:438 arg1 technology 461:470 recently developed high-throughput UPLC technology 421:470 recently developed high-throughput UPLC technology 421:470 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 7 87 gly core-fucosylation 1075:1091 arg1 glycans 1104:1110 neutral glycans 1096:1110 neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans 1096:1178 RESULTS We have found that colorectal cancer associates with decrease in IgG galactosylation, IgG sialylation and increase in core-fucosylation of neutral glycans with concurrent decrease of core-fucosylation of sialylated glycans. 26831718 3 88 theme glycosylation 480:492 arg1 analysis 494:501 IgG glycosylation analysis 476:501 IgG glycosylation analysis 476:501 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 0 89 theme Colorectal 15:24 arg1 Cancer 26:31 Colorectal Cancer 15:31 Colorectal Cancer 15:31 IgG Glycome in Colorectal Cancer. 26831718 3 90 theme glycome 520:526 arg1 composition 528:538 IgG glycome composition 516:538 IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls 516:603 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 9 91 from set 1457:1459 arg1 significant 1432:1442 significant 1432:1442 significant 1432:1442 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 6 92 theme cancer 941:946 arg1 diagnosis 917:925 initial diagnosis 909:925 initial diagnosis of colorectal cancer 909:946 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 2 93 from differences 282:292 arg1 dynamics 301:308 the dynamics 297:308 the dynamics of changes in IgG glycosylation on colorectal cancer 297:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 10 94 theme functional 1540:1549 arg1 relevance 1551:1559 the functional relevance 1536:1559 the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs 1536:1657 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 6 95 theme IgG 834:836 arg1 composition 846:856 IgG glycome composition 834:856 IgG glycome composition 834:856 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 3 96 with controls 596:603 arg1 cancer 572:577 colorectal cancer 561:577 colorectal cancer 561:577 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 3 97 theme UPLC 456:459 arg1 technology 461:470 recently developed high-throughput UPLC technology 421:470 recently developed high-throughput UPLC technology 421:470 EXPERIMENTAL DESIGN Using recently developed high-throughput UPLC technology for IgG glycosylation analysis, we analyzed IgG glycome composition in 760 patients with colorectal cancer and 538 matching controls. 26831718 9 98 from significant 1432:1442 arg1 samples 1464:1470 samples 1464:1470 samples collected before the initial diagnosis 1464:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 9 98 from significant 1432:1442 arg1 set 1457:1459 the small set 1447:1459 the small set of samples collected before the initial diagnosis 1447:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 6 99 theme initial 909:915 arg1 diagnosis 917:925 initial diagnosis 909:925 initial diagnosis of colorectal cancer 909:946 Furthermore, IgG glycome composition was analyzed in 39 plasma samples collected before initial diagnosis of colorectal cancer. 26831718 10 100 theme colorectal 1861:1870 arg1 cancer 1872:1877 colorectal cancer 1861:1877 colorectal cancer 1861:1877 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 8 101 dep increased 1329:1337 arg1 AUC 1378:1380 AUC = 0.755 1378:1388 AUC = 0.755 1378:1388 Although a model based on age and sex did not show discriminative power (AUC = 0.499), the addition of glycan variables into the model considerably increased the discriminative power of the model (AUC = 0.755). 26831718 2 102 theme colorectal 345:354 arg1 cancer 356:361 colorectal cancer 345:361 colorectal cancer 345:361 Because of technological limitations, the effects of highly heritable individual variations and the differences in the dynamics of changes in IgG glycosylation on colorectal cancer were never investigated before. 26831718 1 103 theme PURPOSE 34:40 arg1 glycosylation 54:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation 34:66 PURPOSE Alternative glycosylation has significant structural and functional consequences on IgG and consequently also on cancer immunosurveillance. 26831718 10 104 theme clinical 1620:1627 arg1 efficacy 1629:1636 clinical efficacy 1620:1636 clinical efficacy 1620:1636 CONCLUSIONS Considering the functional relevance of IgG glycosylation for both tumor immunosurveillance and clinical efficacy of therapy with mAbs, individual variation in IgG glycosylation may turn out to be important for prediction of disease course or the choice of therapy, thus warranting further, more detailed studies of IgG glycosylation in colorectal cancer. 26831718 9 105 theme small 1451:1455 arg1 samples 1464:1470 samples 1464:1470 samples collected before the initial diagnosis 1464:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 26831718 9 105 theme small 1451:1455 arg1 set 1457:1459 the small set 1447:1459 the small set of samples collected before the initial diagnosis 1447:1509 However, none of these differences were significant in the small set of samples collected before the initial diagnosis. 27145274 0 0 theme new 94:96 arg1 properties 109:118 new structural properties 94:118 new structural properties of the molecule 94:134 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 2 1 theme silver 430:435 arg1 stain 437:441 silver stain 430:441 silver stain 430:441 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 5 2 theme new 950:952 arg1 structure 970:978 new basic molecular structure 950:978 new basic molecular structure of IgG 950:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 3 3 theme N- 638:639 arg1 oligosaccharide 648:662 N- linked oligosaccharide 638:662 N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc 638:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 5 4 contain have 993:996 arg2 implications 998:1009 implications 998:1009 implications 998:1009 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 5 4 contain have 993:996 arg1 discoveries 935:945 The discoveries 931:945 The discoveries of new basic molecular structure of IgG 931:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 4 5 theme IgG 852:854 arg1 level 843:847 detectable level 832:847 detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously 832:928 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 5 6 theme structure 970:978 arg1 discoveries 935:945 The discoveries 931:945 The discoveries of new basic molecular structure of IgG 931:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 5 7 with properties 1045:1054 arg1 applications 1104:1115 clinical applications 1095:1115 clinical applications 1095:1115 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 3 8 located located 711:717 arg2 oligosaccharide 648:662 N- linked oligosaccharide 638:662 N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc 638:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 8 located located 711:717 arg1 contrast 628:635 contrast 628:635 contrast 628:635 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 8 located located 711:717 arg1 inside 726:731 the inside 722:731 the inside of the natural protein 722:754 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 2 9 theme lectin 461:466 arg1 techniques 474:483 lectin stain techniques 461:483 lectin stain techniques 461:483 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 0 10 theme structural 98:107 arg1 properties 109:118 new structural properties 94:118 new structural properties of the molecule 94:134 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 1 11 theme arms 314:317 arg1 one 291:293 one 291:293 one 291:293 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 11 theme arms 314:317 arg1 arms 314:317 the two F(ab')2 arms 298:317 the two F(ab')2 arms 298:317 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 2 12 theme Fab 524:526 arg1 fragment 528:535 Fab fragment 524:535 Fab fragment 524:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 1 13 attach attached 279:286 arg2 oligosaccharide 263:277 oligosaccharide 263:277 oligosaccharide attached to one of the two F(ab')2 arms 263:317 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 13 attach attached 279:286 arg1 one 291:293 one 291:293 one 291:293 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 13 attach attached 279:286 arg1 arms 314:317 the two F(ab')2 arms 298:317 the two F(ab')2 arms 298:317 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 2 14 attach attached 512:519 arg2 oligosaccharide 496:510 N- linked oligosaccharide 486:510 N- linked oligosaccharide attached to Fab fragment 486:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 2 14 attach attached 512:519 arg1 fragment 528:535 Fab fragment 524:535 Fab fragment 524:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 4 15 theme detectable 832:841 arg1 level 843:847 detectable level 832:847 detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously 832:928 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 3 16 theme linked 641:646 arg1 oligosaccharide 648:662 N- linked oligosaccharide 638:662 N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc 638:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 1 17 theme separate 203:210 arg1 IgG 241:243 IgG 241:243 IgG 241:243 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 17 theme separate 203:210 arg1 G 238:238 separate asymmetric Immunoglobulin G 203:238 separate asymmetric Immunoglobulin G (IgG) 203:244 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 2 18 theme Western 444:450 arg1 blot 452:455 Western blot 444:455 Western blot 444:455 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 4 19 theme F 866:866 arg1 arms 874:877 both F(ab')2 arms 861:877 both F(ab')2 arms glycosylated that has not been reported previously 861:928 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 3 20 from contrast 628:635 arg1 located 711:717 located 711:717 located 711:717 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 1 21 theme symmetric 325:333 arg1 IgG 335:337 symmetric IgG 325:337 symmetric IgG with no glycan attached to Fab fragments 325:378 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 5 22 with function 1032:1039 arg1 applications 1104:1115 clinical applications 1095:1115 clinical applications 1095:1115 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 3 23 link linked 641:646 arg1 oligosaccharide 648:662 N- linked oligosaccharide 638:662 N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc 638:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 4 24 with level 843:847 arg1 arms 874:877 both F(ab')2 arms 861:877 both F(ab')2 arms glycosylated that has not been reported previously 861:928 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 0 25 theme glycosylated 21:32 arg1 G 49:49 Fab glycosylated immunoglobulin G 17:49 Fab glycosylated immunoglobulin G 17:49 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 3 26 theme asparagine 676:685 arg1 297 693:695 asparagine (Asn) 297 676:695 asparagine (Asn) 297 of IgG Fc 676:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 2 27 theme N- 486:487 arg1 oligosaccharide 496:510 N- linked oligosaccharide 486:510 N- linked oligosaccharide attached to Fab fragment 486:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 2 28 theme protein 590:596 arg1 surface 575:581 the surface 571:581 the surface of the protein 571:596 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 2 29 link linked 489:494 arg1 oligosaccharide 496:510 N- linked oligosaccharide 486:510 N- linked oligosaccharide attached to Fab fragment 486:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 0 30 theme Fab 17:19 arg1 G 49:49 Fab glycosylated immunoglobulin G 17:49 Fab glycosylated immunoglobulin G 17:49 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 5 31 theme molecular 960:968 arg1 structure 970:978 new basic molecular structure 950:978 new basic molecular structure of IgG 950:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 0 32 theme molecule 127:134 arg1 properties 109:118 new structural properties 94:118 new structural properties of the molecule 94:134 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 1 33 with IgG 335:337 arg1 glycan 347:352 no glycan 344:352 no glycan attached to Fab fragments 344:378 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 34 attach attached 354:361 arg2 glycan 347:352 no glycan 344:352 no glycan attached to Fab fragments 344:378 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 34 attach attached 354:361 arg1 fragments 370:378 Fab fragments 366:378 Fab fragments 366:378 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 0 35 theme G 49:49 arg1 Fractionation 0:12 Fractionation 0:12 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography 0:84 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 1 36 theme asymmetric 212:221 arg1 IgG 241:243 IgG 241:243 IgG 241:243 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 36 theme asymmetric 212:221 arg1 G 238:238 separate asymmetric Immunoglobulin G 203:238 separate asymmetric Immunoglobulin G (IgG) 203:244 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 0 37 theme immunoglobulin 34:47 arg1 G 49:49 Fab glycosylated immunoglobulin G 17:49 Fab glycosylated immunoglobulin G 17:49 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 5 38 theme IgG 983:985 arg1 structure 970:978 new basic molecular structure 950:978 new basic molecular structure of IgG 950:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 2 39 theme affinity 405:412 arg1 chromatography 414:427 affinity chromatography 405:427 affinity chromatography 405:427 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 1 40 theme Immunoglobulin 223:236 arg1 IgG 241:243 IgG 241:243 IgG 241:243 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 40 theme Immunoglobulin 223:236 arg1 G 238:238 separate asymmetric Immunoglobulin G 203:238 separate asymmetric Immunoglobulin G (IgG) 203:244 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 0 41 theme concanavalin 56:67 arg1 chromatography 71:84 concanavalin A chromatography 56:84 concanavalin A chromatography 56:84 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 5 42 dep function 1032:1039 arg1 the 1028:1030 the 1028:1030 the 1028:1030 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 2 43 theme stain 468:472 arg1 techniques 474:483 lectin stain techniques 461:483 lectin stain techniques 461:483 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 1 44 theme F 306:306 arg1 arms 314:317 the two F(ab')2 arms 298:317 the two F(ab')2 arms 298:317 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 3 45 from located 711:717 arg1 contrast 628:635 contrast 628:635 contrast 628:635 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 45 from located 711:717 arg1 inside 726:731 the inside 722:731 the inside of the natural protein 722:754 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 4 46 gly glycosylated 879:890 arg1 arms 874:877 both F(ab')2 arms 861:877 both F(ab')2 arms glycosylated that has not been reported previously 861:928 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 0 47 gly glycosylated 21:32 arg1 G 49:49 Fab glycosylated immunoglobulin G 17:49 Fab glycosylated immunoglobulin G 17:49 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 3 48 theme natural 740:746 arg1 protein 748:754 the natural protein 736:754 the natural protein 736:754 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 49 attach attached 664:671 arg2 oligosaccharide 648:662 N- linked oligosaccharide 638:662 N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc 638:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 49 attach attached 664:671 arg1 297 693:695 asparagine (Asn) 297 676:695 asparagine (Asn) 297 of IgG Fc 676:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 1 50 used used 195:198 arg2 chromatography 159:172 Concanavalin A (ConA) chromatography 137:172 Concanavalin A (ConA) chromatography 137:172 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 0 51 with Fractionation 0:12 arg1 chromatography 71:84 concanavalin A chromatography 56:84 concanavalin A chromatography 56:84 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 1 52 theme Concanavalin 137:148 arg1 ConA 153:156 ConA 153:156 ConA 153:156 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 52 theme Concanavalin 137:148 arg1 A 150:150 Concanavalin A 137:150 Concanavalin A (ConA) chromatography 137:172 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 3 53 theme protein 748:754 arg1 inside 726:731 the inside 722:731 the inside of the natural protein 722:754 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 3 54 theme Fc 704:705 arg1 297 693:695 asparagine (Asn) 297 676:695 asparagine (Asn) 297 of IgG Fc 676:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 2 55 theme linked 489:494 arg1 oligosaccharide 496:510 N- linked oligosaccharide 486:510 N- linked oligosaccharide attached to Fab fragment 486:535 In this study, applying affinity chromatography, silver stain, Western blot and lectin stain techniques, N- linked oligosaccharide attached to Fab fragment was demonstrated to be exposed on the surface of the protein and be accessible by ConA. 27145274 1 56 theme Fab 366:368 arg1 fragments 370:378 Fab fragments 366:378 Fab fragments 366:378 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 57 contain possesses 253:261 arg2 oligosaccharide 263:277 oligosaccharide 263:277 oligosaccharide attached to one of the two F(ab')2 arms 263:317 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 57 contain possesses 253:261 arg1 IgG 241:243 IgG 241:243 IgG 241:243 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 57 contain possesses 253:261 arg1 G 238:238 separate asymmetric Immunoglobulin G 203:238 separate asymmetric Immunoglobulin G (IgG) 203:244 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 1 58 theme A 150:150 arg1 chromatography 159:172 Concanavalin A (ConA) chromatography 137:172 Concanavalin A (ConA) chromatography 137:172 Concanavalin A (ConA) chromatography has been extensively used to separate asymmetric Immunoglobulin G (IgG), which possesses oligosaccharide attached to one of the two F(ab')2 arms, from symmetric IgG with no glycan attached to Fab fragments. 27145274 5 59 theme immune 1074:1079 arg1 molecule 1081:1088 this important immune molecule 1059:1088 this important immune molecule 1059:1088 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 4 60 theme asymmetric 801:810 arg1 IgG 812:814 asymmetric IgG 801:814 asymmetric IgG 801:814 In addition to asymmetric IgG, there are also detectable level of IgG with both F(ab')2 arms glycosylated that has not been reported previously. 27145274 3 61 from inside 726:731 arg1 located 711:717 located 711:717 located 711:717 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 27145274 5 62 theme molecule 1081:1088 arg1 properties 1045:1054 properties 1045:1054 properties 1045:1054 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 5 62 theme molecule 1081:1088 arg1 function 1032:1039 function 1032:1039 function 1032:1039 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 0 63 theme A 69:69 arg1 chromatography 71:84 concanavalin A chromatography 56:84 concanavalin A chromatography 56:84 Fractionation of Fab glycosylated immunoglobulin G with concanavalin A chromatography unveils new structural properties of the molecule. 27145274 5 64 theme basic 954:958 arg1 structure 970:978 new basic molecular structure 950:978 new basic molecular structure of IgG 950:985 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 5 65 theme important 1064:1072 arg1 molecule 1081:1088 this important immune molecule 1059:1088 this important immune molecule 1059:1088 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 5 66 theme clinical 1095:1102 arg1 applications 1104:1115 clinical applications 1095:1115 clinical applications 1095:1115 The discoveries of new basic molecular structure of IgG would have implications in understanding the function and properties of this important immune molecule with clinical applications. 27145274 3 67 theme IgG 700:702 arg1 Fc 704:705 IgG Fc 700:705 IgG Fc 700:705 In contrast, N- linked oligosaccharide attached to asparagine (Asn) 297 of IgG Fc was located in the inside of the natural protein and was inaccessible by ConA. 29237727 0 0 from Micro-heterogeneity 16:34 arg1 Glycoproteins 39:51 Glycoproteins 39:51 Glycoproteins of Arabidopsis 39:66 N-linked Glycan Micro-heterogeneity in Glycoproteins of Arabidopsis. 29237727 5 1 theme 492 703:705 arg1 N-glycosites 707:718 492 N-glycosites 703:718 492 N-glycosites 703:718 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 7 2 from N-glycoproteins 1215:1229 arg1 present 1204:1210 present 1204:1210 present 1204:1210 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 4 3 from membranes 663:671 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 4 4 theme mass 543:546 arg1 spectrometry 548:559 hydrophilic enrichment, high-resolution tandem mass spectrometry 496:559 spectrometry 548:559 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 5 5 theme N-glycosites 707:718 arg1 total 694:698 A total 692:698 A total of 492 N-glycosites 692:718 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 4 6 theme Arabidopsis 619:629 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 4 7 theme tandem 536:541 arg1 spectrometry 548:559 hydrophilic enrichment, high-resolution tandem mass spectrometry 496:559 spectrometry 548:559 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 7 8 theme Arabidopsis 1234:1244 arg1 N-glycoproteins 1215:1229 N-glycoproteins 1215:1229 N-glycoproteins of Arabidopsis 1215:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 3 9 theme advanced 406:413 arg1 spectrometry 420:431 advanced mass spectrometry 406:431 advanced mass spectrometry 406:431 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 4 10 with spectrometry 548:559 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 4 11 theme microsomal 652:661 arg1 membranes 663:671 microsomal membranes 652:671 microsomal membranes of aerial tissues 652:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 3 12 theme few 381:383 arg1 studies 385:391 few studies 381:391 few studies 381:391 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 5 13 gly N-glycopeptides 837:851 arg2 N-glycopeptides 837:851 1110 N-glycopeptides 832:851 1110 N-glycopeptides 832:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 7 14 gly N-glycoproteins 1215:1229 arg1 N-glycoproteins 1215:1229 N-glycoproteins 1215:1229 N-glycoproteins of Arabidopsis 1215:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 0 15 link N-linked 0:7 arg1 Glycan 9:14 N-linked Glycan 0:14 N-linked Glycan 0:14 N-linked Glycan Micro-heterogeneity in Glycoproteins of Arabidopsis. 29237727 1 16 theme conserved 187:195 arg1 structure 202:210 a relatively conserved core structure 174:210 a relatively conserved core structure between fungi, animals and plants 174:244 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 5 17 gly N-glycosites 707:718 arg2 N-glycosites 707:718 492 N-glycosites 703:718 492 N-glycosites 703:718 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 1 18 contain has 170:172 arg1 N-glycosylation 69:83 N-glycosylation 69:83 N-glycosylation 69:83 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 1 18 contain has 170:172 arg2 structure 202:210 a relatively conserved core structure 174:210 a relatively conserved core structure between fungi, animals and plants 174:244 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 2 19 theme N-glycans 278:286 arg1 biosynthesis 262:273 the biosynthesis 258:273 the biosynthesis of N-glycans 258:286 In plants, the biosynthesis of N-glycans has been extensively studied with all the major biosynthetic enzymes characterized. 29237727 1 20 theme core 197:200 arg1 structure 202:210 a relatively conserved core structure 174:210 a relatively conserved core structure between fungi, animals and plants 174:244 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 4 21 theme hydrophilic 496:506 arg1 spectrometry 548:559 hydrophilic enrichment, high-resolution tandem mass spectrometry 496:559 spectrometry 548:559 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 7 22 theme present 1204:1210 arg1 micro-heterogeneity 1184:1202 the micro-heterogeneity 1180:1202 the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1180:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 4 23 gly N-glycopeptides 631:645 arg1 membranes 663:671 microsomal membranes 652:671 microsomal membranes of aerial tissues 652:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 4 23 gly N-glycopeptides 631:645 arg2 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 6 24 gly N-glycopeptides 917:931 arg1 xylt 950:953 xylt 950:953 xylt 950:953 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 6 24 gly N-glycopeptides 917:931 arg1 mutant 942:947 the mutant 938:947 the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway 938:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 6 24 gly N-glycopeptides 917:931 arg2 N-glycopeptides 917:931 N-glycopeptides 917:931 N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway 917:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 3 25 theme intact 444:449 arg1 N-glycopeptides 457:471 profile intact plant N-glycopeptides 436:471 profile intact plant N-glycopeptides 436:471 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 0 26 theme N-linked 0:7 arg1 Glycan 9:14 N-linked Glycan 0:14 N-linked Glycan 0:14 N-linked Glycan Micro-heterogeneity in Glycoproteins of Arabidopsis. 29237727 4 27 theme fragmentation 594:606 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 6 28 from mutant 942:947 arg1 N-glycopeptides 917:931 N-glycopeptides 917:931 N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway 917:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 3 29 theme plant 451:455 arg1 N-glycopeptides 457:471 profile intact plant N-glycopeptides 436:471 profile intact plant N-glycopeptides 436:471 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 4 30 theme triggered 584:592 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 1 31 from modifications 138:150 arg1 eukaryotes 155:164 eukaryotes 155:164 eukaryotes 155:164 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 1 32 theme common 104:109 arg1 modifications 138:150 the most common protein post-translational modifications 95:150 the most common protein post-translational modifications in eukaryotes 95:164 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 4 33 theme profile 611:617 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 5 34 theme extensive 771:779 arg1 heterogeneity 801:813 extensive N-glycan structural heterogeneity 771:813 extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 771:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 1 35 theme protein 111:117 arg1 modifications 138:150 the most common protein post-translational modifications 95:150 the most common protein post-translational modifications in eukaryotes 95:164 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 6 36 theme β-1,2-xylosyltransferase 959:982 arg1 xylt 950:953 xylt 950:953 xylt 950:953 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 6 36 theme β-1,2-xylosyltransferase 959:982 arg1 mutant 942:947 the mutant 938:947 the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway 938:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 5 37 theme N-glycan 781:788 arg1 heterogeneity 801:813 extensive N-glycan structural heterogeneity 771:813 extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 771:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 7 38 attach present 1204:1210 arg2 micro-heterogeneity 1184:1202 the micro-heterogeneity 1180:1202 the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1180:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 7 38 attach present 1204:1210 arg1 N-glycoproteins 1215:1229 N-glycoproteins 1215:1229 N-glycoproteins of Arabidopsis 1215:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 3 39 theme profile 436:442 arg1 N-glycopeptides 457:471 profile intact plant N-glycopeptides 436:471 profile intact plant N-glycopeptides 436:471 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 5 40 theme structural 790:799 arg1 heterogeneity 801:813 extensive N-glycan structural heterogeneity 771:813 extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 771:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 4 41 dep N-glycopeptides 631:645 arg1 to 608:609 to 608:609 to 608:609 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 7 42 theme N-glycopeptides 1120:1134 arg1 collection 1094:1103 the most comprehensive and unbiased collection 1058:1103 the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1058:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 6 43 theme biosynthetic 1011:1022 arg1 pathway 1024:1030 the N-glycan biosynthetic pathway 998:1030 the N-glycan biosynthetic pathway 998:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 3 44 theme mass 415:418 arg1 spectrometry 420:431 advanced mass spectrometry 406:431 advanced mass spectrometry 406:431 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 7 45 gly N-glycopeptides 1120:1134 arg2 N-glycopeptides 1120:1134 Arabidopsis N-glycopeptides 1108:1134 Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1108:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 7 46 from present 1204:1210 arg1 N-glycoproteins 1215:1229 N-glycoproteins 1215:1229 N-glycoproteins of Arabidopsis 1215:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 4 47 theme enrichment 508:517 arg1 spectrometry 548:559 hydrophilic enrichment, high-resolution tandem mass spectrometry 496:559 spectrometry 548:559 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 1 48 theme post-translational 119:136 arg1 modifications 138:150 the most common protein post-translational modifications 95:150 the most common protein post-translational modifications in eukaryotes 95:164 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 0 49 theme Arabidopsis 56:66 arg1 Glycoproteins 39:51 Glycoproteins 39:51 Glycoproteins of Arabidopsis 39:66 N-linked Glycan Micro-heterogeneity in Glycoproteins of Arabidopsis. 29237727 5 50 theme 324 741:743 arg1 proteins 757:764 324 Arabidopsis proteins 741:764 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 741:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 1 51 theme modifications 138:150 arg1 one 88:90 one 88:90 one 88:90 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 1 51 theme modifications 138:150 arg1 modifications 138:150 the most common protein post-translational modifications 95:150 the most common protein post-translational modifications in eukaryotes 95:164 N-glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. 29237727 4 52 theme complementary 566:578 arg1 N-glycopeptides 631:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides 566:645 complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues 566:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 5 53 theme Arabidopsis 745:755 arg1 proteins 757:764 324 Arabidopsis proteins 741:764 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 741:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 8 54 with ProteomeXchange 1270:1284 arg1 identifier 1291:1300 identifier PXD006270 1291:1310 identifier PXD006270 1291:1310 Data are available via ProteomeXchange with identifier PXD006270. 29237727 7 55 theme Arabidopsis 1108:1118 arg1 N-glycopeptides 1120:1134 Arabidopsis N-glycopeptides 1108:1134 Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1108:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 2 56 theme biosynthetic 336:347 arg1 enzymes 349:355 all the major biosynthetic enzymes 322:355 all the major biosynthetic enzymes characterized 322:369 In plants, the biosynthesis of N-glycans has been extensively studied with all the major biosynthetic enzymes characterized. 29237727 7 57 theme unsurpassed 1149:1159 arg1 level 1161:1165 an unsurpassed level 1146:1165 an unsurpassed level of detail 1146:1175 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 3 58 gly N-glycopeptides 457:471 arg2 N-glycopeptides 457:471 profile intact plant N-glycopeptides 436:471 profile intact plant N-glycopeptides 436:471 However, few studies have applied advanced mass spectrometry to profile intact plant N-glycopeptides. 29237727 7 59 theme unbiased 1085:1092 arg1 collection 1094:1103 the most comprehensive and unbiased collection 1058:1103 the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1058:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 2 60 theme major 330:334 arg1 enzymes 349:355 all the major biosynthetic enzymes 322:355 all the major biosynthetic enzymes characterized 322:369 In plants, the biosynthesis of N-glycans has been extensively studied with all the major biosynthetic enzymes characterized. 29237727 6 61 theme approach 890:897 arg1 precision 873:881 the precision 869:881 the precision of the approach 869:897 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 6 62 theme N-glycan 1002:1009 arg1 pathway 1024:1030 the N-glycan biosynthetic pathway 998:1030 the N-glycan biosynthetic pathway 998:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 5 63 with proteins 757:764 arg1 heterogeneity 801:813 extensive N-glycan structural heterogeneity 771:813 extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides 771:851 A total of 492 N-glycosites were identified from 324 Arabidopsis proteins with extensive N-glycan structural heterogeneity revealed through 1110 N-glycopeptides. 29237727 4 64 theme tissues 683:689 arg1 membranes 663:671 microsomal membranes 652:671 microsomal membranes of aerial tissues 652:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 6 65 from enzyme 988:993 arg1 pathway 1024:1030 the N-glycan biosynthetic pathway 998:1030 the N-glycan biosynthetic pathway 998:1030 To demonstrate the precision of the approach, we also profiled N-glycopeptides from the mutant (xylt) of β-1,2-xylosyltransferase, an enzyme in the N-glycan biosynthetic pathway. 29237727 4 66 theme high-resolution 520:534 arg1 spectrometry 548:559 hydrophilic enrichment, high-resolution tandem mass spectrometry 496:559 spectrometry 548:559 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 29237727 7 67 theme detail 1170:1175 arg1 level 1161:1165 an unsurpassed level 1146:1165 an unsurpassed level of detail 1146:1175 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 7 68 theme comprehensive 1067:1079 arg1 collection 1094:1103 the most comprehensive and unbiased collection 1058:1103 the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis 1058:1244 This analysis represents the most comprehensive and unbiased collection of Arabidopsis N-glycopeptides revealing an unsurpassed level of detail on the micro-heterogeneity present in N-glycoproteins of Arabidopsis. 29237727 4 69 theme aerial 676:681 arg1 tissues 683:689 aerial tissues 676:689 aerial tissues 676:689 In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N-glycopeptides from microsomal membranes of aerial tissues. 25501957 8 0 theme Glc 1184:1186 arg1 α-1 1170:1172 Glc α-1 1166:1172 Glc α-1 1166:1172 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 0 theme Glc 1184:1186 arg1 α-1 1188:1190 Glc α-1 1184:1190 Glc α-1 1184:1190 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 5 1 contain has 767:769 arg2 property 798:805 unique transfructosylating property 771:805 unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS 771:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 5 1 contain has 767:769 arg1 fructosidases 700:712 fructosidases 700:712 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA 700:765 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 2 2 theme high 397:400 arg1 concentrations 402:415 high concentrations 397:415 high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry 397:505 These enzymes also exhibit transglycosylation activity when they function with high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry. 25501957 1 3 theme units 287:291 arg1 hydrolysis 254:263 the hydrolysis 250:263 the hydrolysis of terminal fructosyl units from various substrates 250:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 9 4 theme total 1572:1576 arg1 activity 1578:1585 total activity 1572:1585 total activity 1572:1585 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 3 5 theme Aspergillus 588:598 arg1 oryzae 600:605 Aspergillus oryzae 588:605 Aspergillus oryzae FS4 as a monomeric glycoprotein 588:637 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 1 6 theme various 298:304 arg1 substrates 306:315 various substrates 298:315 various substrates 298:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 6 7 theme BfrA 910:913 arg1 sequence 879:886 The coding sequence 868:886 The coding sequence (bfrAFS4, 1.86 kb) of BfrA 868:913 The coding sequence (bfrAFS4, 1.86 kb) of BfrA was amplified and expressed in Escherichia coli and Pichia pastoris. 25501957 6 7 theme BfrA 910:913 arg1 bfrAFS4 889:895 bfrAFS4 889:895 bfrAFS4 889:895 The coding sequence (bfrAFS4, 1.86 kb) of BfrA was amplified and expressed in Escherichia coli and Pichia pastoris. 25501957 9 8 from temperatures 1460:1471 arg1 thermostable 1444:1455 thermostable 1444:1455 thermostable 1444:1455 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 11 9 theme neolevan-type 1951:1963 arg1 FOS 1965:1967 neolevan-type FOS 1951:1967 neolevan-type FOS 1951:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 9 10 gly unglycosylated 1242:1255 arg1 the 1329:1331 the 1329:1331 the 1329:1331 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 10 gly unglycosylated 1242:1255 arg1 E.BfrA 1281:1286 E.BfrA 1281:1286 E.BfrA 1281:1286 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 10 gly unglycosylated 1242:1255 arg1 P. 1333:1334 P. 1333:1334 P. 1333:1334 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 10 gly unglycosylated 1242:1255 arg1 BfrA 1275:1278 the unglycosylated E. coli-expressed BfrA 1238:1278 the unglycosylated E. coli-expressed BfrA (E.BfrA) 1238:1287 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 2 11 theme transglycosylation 345:362 arg1 activity 364:371 transglycosylation activity 345:371 transglycosylation activity 345:371 These enzymes also exhibit transglycosylation activity when they function with high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry. 25501957 4 12 theme Aspergillus 683:693 arg1 spp 695:697 the most extensively studied Aspergillus spp 654:697 the most extensively studied Aspergillus spp 654:697 Compared with the most extensively studied Aspergillus spp. 25501957 3 13 theme high 537:540 arg1 activity 561:568 high transglycosylation activity 537:568 high transglycosylation activity 537:568 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 1 14 theme enzymes 228:234 arg1 enzymes 228:234 enzymes 228:234 enzymes 228:234 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 14 theme enzymes 228:234 arg1 group 219:223 a widespread group 206:223 a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates 206:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 14 theme enzymes 228:234 arg1 β-Fructosidases 186:200 β-Fructosidases 186:200 β-Fructosidases 186:200 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 3 15 gly glycoprotein 626:637 arg1 glycoprotein 626:637 a monomeric glycoprotein 614:637 a monomeric glycoprotein 614:637 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 0 16 theme levan- 137:142 arg1 fructooligosaccharides 162:183 levan- and neolevan-type fructooligosaccharides 137:183 levan- and neolevan-type fructooligosaccharides 137:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 11 17 theme transfructosylating 1893:1911 arg1 capability 1913:1922 unique transfructosylating capability 1886:1922 unique transfructosylating capability of synthesizing levan- and neolevan-type FOS 1886:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 7 18 theme hydrolyzation 1052:1064 arg1 activities 1066:1075 transfructosylation and hydrolyzation activities 1028:1075 transfructosylation and hydrolyzation activities 1028:1075 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 0 19 from analysis 61:68 arg1 FS4 120:122 Aspergillus oryzae FS4 101:122 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 10 20 gly N-glycosylation 1726:1740 arg1 optima 1789:1794 optima activity 1789:1803 optima activity 1789:1803 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 10 20 gly N-glycosylation 1726:1740 arg1 enzyme 1760:1765 enzyme thermal stability 1760:1783 enzyme thermal stability 1760:1783 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 0 21 theme neolevan-type 148:160 arg1 fructooligosaccharides 162:183 levan- and neolevan-type fructooligosaccharides 137:183 levan- and neolevan-type fructooligosaccharides 137:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 9 22 theme E. 1257:1258 arg1 the 1329:1331 the 1329:1331 the 1329:1331 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 22 theme E. 1257:1258 arg1 E.BfrA 1281:1286 E.BfrA 1281:1286 E.BfrA 1281:1286 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 22 theme E. 1257:1258 arg1 P. 1333:1334 P. 1333:1334 P. 1333:1334 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 22 theme E. 1257:1258 arg1 BfrA 1275:1278 the unglycosylated E. coli-expressed BfrA 1238:1278 the unglycosylated E. coli-expressed BfrA (E.BfrA) 1238:1287 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 7 23 theme transfructosylation 1028:1046 arg1 activities 1066:1075 transfructosylation and hydrolyzation activities 1028:1075 transfructosylation and hydrolyzation activities 1028:1075 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 9 24 from 55°C. 1511:1515 arg1 activity 1499:1506 a maximum activity 1489:1506 a maximum activity at 55°C. Using sucrose as substrate 1489:1542 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 25 dep 50°C 1479:1482 arg1 up 1473:1474 up 1473:1474 up 1473:1474 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 7 26 theme broad 1082:1086 arg1 specificity 1098:1108 broad substrate specificity 1082:1108 broad substrate specificity 1082:1108 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 5 27 link -linked 855:861 arg1 FOS 863:865 neolevan-type β-(2-6)-linked FOS 834:865 neolevan-type β-(2-6)-linked FOS 834:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 0 28 from cloning 14:20 arg1 FS4 120:122 Aspergillus oryzae FS4 101:122 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 2 29 used used 438:441 arg2 sucrose 420:426 sucrose 420:426 sucrose 420:426 These enzymes also exhibit transglycosylation activity when they function with high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry. 25501957 0 30 from Purification 0:11 arg1 FS4 120:122 Aspergillus oryzae FS4 101:122 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 31 theme β-fructosidase 81:94 arg1 cloning 14:20 cloning 14:20 cloning 14:20 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 31 theme β-fructosidase 81:94 arg1 Purification 0:11 Purification 0:11 Purification 0:11 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 31 theme β-fructosidase 81:94 arg1 analysis 61:68 N-glycosylation analysis 45:68 N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 45:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 31 theme β-fructosidase 81:94 arg1 characterization 23:38 characterization 23:38 characterization 23:38 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 10 32 theme putative 1662:1669 arg1 sites 1687:1691 10 of 13 putative N-glycosylation sites 1653:1691 10 of 13 putative N-glycosylation sites 1653:1691 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 9 33 dep P.BfrA 1361:1366 arg1 thermostable 1444:1455 thermostable 1444:1455 thermostable 1444:1455 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 33 dep P.BfrA 1361:1366 arg1 stable 1381:1386 stable 1381:1386 stable 1381:1386 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 34 theme pH 1398:1399 arg1 pH 1408:1409 pH 4 to 11 1408:1417 pH 4 to 11 1408:1417 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 34 theme pH 1398:1399 arg1 range 1401:1405 a wide pH range 1391:1405 a wide pH range (pH 4 to 11) 1391:1418 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 35 theme maximum 1491:1497 arg1 activity 1499:1506 a maximum activity 1489:1506 a maximum activity at 55°C. Using sucrose as substrate 1489:1542 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 0 36 from characterization 23:38 arg1 FS4 120:122 Aspergillus oryzae FS4 101:122 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 5 37 theme -linked 855:861 arg1 FOS 863:865 neolevan-type β-(2-6)-linked FOS 834:865 neolevan-type β-(2-6)-linked FOS 834:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 10 38 gly glycosylated 1698:1709 arg2 N.BfrA 1714:1719 N.BfrA 1714:1719 N.BfrA 1714:1719 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 10 38 gly glycosylated 1698:1709 arg1 N.BfrA 1714:1719 N.BfrA 1714:1719 N.BfrA 1714:1719 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 10 38 gly glycosylated 1698:1709 arg2 sites 1687:1691 10 of 13 putative N-glycosylation sites 1653:1691 10 of 13 putative N-glycosylation sites 1653:1691 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 10 38 gly glycosylated 1698:1709 arg1 sites 1687:1691 10 of 13 putative N-glycosylation sites 1653:1691 10 of 13 putative N-glycosylation sites 1653:1691 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 11 39 theme A. 1858:1859 arg1 fructosidase 1868:1879 a well-characterized A. oryzae fructosidase 1837:1879 a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS 1837:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 9 40 dep pH 1408:1409 arg1 to 1413:1414 to 1413:1414 to 1413:1414 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 5 41 theme β- 848:849 arg1 FOS 863:865 neolevan-type β-(2-6)-linked FOS 834:865 neolevan-type β-(2-6)-linked FOS 834:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 8 42 theme following 1146:1154 arg1 linkages 1156:1163 the following linkages 1142:1163 the following linkages 1142:1163 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 6 43 dep bfrAFS4 889:895 arg1 kb 903:904 1.86 kb 898:904 1.86 kb 898:904 The coding sequence (bfrAFS4, 1.86 kb) of BfrA was amplified and expressed in Escherichia coli and Pichia pastoris. 25501957 10 44 theme thermal 1767:1773 arg1 stability 1775:1783 enzyme thermal stability 1760:1783 enzyme thermal stability 1760:1783 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 6 45 theme coding 872:877 arg1 sequence 879:886 The coding sequence 868:886 The coding sequence (bfrAFS4, 1.86 kb) of BfrA 868:913 The coding sequence (bfrAFS4, 1.86 kb) of BfrA was amplified and expressed in Escherichia coli and Pichia pastoris. 25501957 6 45 theme coding 872:877 arg1 bfrAFS4 889:895 bfrAFS4 889:895 bfrAFS4 889:895 The coding sequence (bfrAFS4, 1.86 kb) of BfrA was amplified and expressed in Escherichia coli and Pichia pastoris. 25501957 9 46 theme Km 1549:1550 arg1 values 1561:1566 the Km and kcat values 1545:1566 values 1561:1566 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 46 theme Km 1549:1550 arg1 mM 1603:1604 37.19±5.28 mM 1592:1604 37.19±5.28 mM 1592:1604 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 11 47 theme well-characterized 1839:1856 arg1 fructosidase 1868:1879 a well-characterized A. oryzae fructosidase 1837:1879 a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS 1837:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 3 48 theme transglycosylation 542:559 arg1 activity 561:568 high transglycosylation activity 537:568 high transglycosylation activity 537:568 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 8 49 theme 2-β 1175:1177 arg1 α-1 1170:1172 Glc α-1 1166:1172 Glc α-1 1166:1172 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 49 theme 2-β 1175:1177 arg1 Fru 1179:1181 2-β Fru 1175:1181 2-β Fru 1175:1181 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 9 50 theme kcat 1556:1559 arg1 values 1561:1566 the Km and kcat values 1545:1566 values 1561:1566 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 50 theme kcat 1556:1559 arg1 mM 1603:1604 37.19±5.28 mM 1592:1604 37.19±5.28 mM 1592:1604 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 51 from range 1401:1405 arg1 stable 1381:1386 stable 1381:1386 stable 1381:1386 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 0 52 from FS4 120:122 arg1 cloning 14:20 cloning 14:20 cloning 14:20 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 52 from FS4 120:122 arg1 characterization 23:38 characterization 23:38 characterization 23:38 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 52 from FS4 120:122 arg1 Purification 0:11 Purification 0:11 Purification 0:11 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 52 from FS4 120:122 arg1 analysis 61:68 N-glycosylation analysis 45:68 N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 45:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 52 from FS4 120:122 arg1 β-fructosidase 81:94 a novel β-fructosidase 73:94 a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 73:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 5 53 link -linked 749:755 arg1 BfrA 762:765 BfrA 762:765 BfrA 762:765 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 1 54 theme fructosyl 277:285 arg1 units 287:291 terminal fructosyl units 268:291 terminal fructosyl units from various substrates 268:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 9 55 dep P. 1333:1334 arg1 the 1290:1292 the 1290:1292 the 1290:1292 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 55 dep P. 1333:1334 arg1 P.BfrA 1361:1366 P.BfrA 1361:1366 P.BfrA 1361:1366 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 10 56 theme N-glycosylation 1671:1685 arg1 sites 1687:1691 10 of 13 putative N-glycosylation sites 1653:1691 10 of 13 putative N-glycosylation sites 1653:1691 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 9 57 theme wide 1393:1396 arg1 pH 1408:1409 pH 4 to 11 1408:1417 pH 4 to 11 1408:1417 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 57 theme wide 1393:1396 arg1 range 1401:1405 a wide pH range 1391:1405 a wide pH range (pH 4 to 11) 1391:1418 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 3 58 with β-fructosidase 510:523 arg1 activity 561:568 high transglycosylation activity 537:568 high transglycosylation activity 537:568 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 1 59 from substrates 306:315 arg1 units 287:291 terminal fructosyl units 268:291 terminal fructosyl units from various substrates 268:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 59 from substrates 306:315 arg1 hydrolysis 254:263 the hydrolysis 250:263 the hydrolysis of terminal fructosyl units from various substrates 250:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 60 theme widespread 208:217 arg1 enzymes 228:234 enzymes 228:234 enzymes 228:234 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 60 theme widespread 208:217 arg1 group 219:223 a widespread group 206:223 a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates 206:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 1 60 theme widespread 208:217 arg1 β-Fructosidases 186:200 β-Fructosidases 186:200 β-Fructosidases 186:200 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 8 61 theme Glc 1206:1208 arg1 α-1 1210:1212 Glc α-1 1206:1212 Glc α-1 1206:1212 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 61 theme Glc 1206:1208 arg1 α-1 1170:1172 Glc α-1 1166:1172 Glc α-1 1166:1172 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 4 62 theme studied 675:681 arg1 spp 695:697 the most extensively studied Aspergillus spp 654:697 the most extensively studied Aspergillus spp 654:697 Compared with the most extensively studied Aspergillus spp. 25501957 11 63 theme unique 1886:1891 arg1 capability 1913:1922 unique transfructosylating capability 1886:1922 unique transfructosylating capability of synthesizing levan- and neolevan-type FOS 1886:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 7 64 theme substrate 1088:1096 arg1 specificity 1098:1108 broad substrate specificity 1082:1108 broad substrate specificity 1082:1108 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 9 65 theme unglycosylated 1242:1255 arg1 the 1329:1331 the 1329:1331 the 1329:1331 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 65 theme unglycosylated 1242:1255 arg1 E.BfrA 1281:1286 E.BfrA 1281:1286 E.BfrA 1281:1286 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 65 theme unglycosylated 1242:1255 arg1 P. 1333:1334 P. 1333:1334 P. 1333:1334 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 65 theme unglycosylated 1242:1255 arg1 BfrA 1275:1278 the unglycosylated E. coli-expressed BfrA 1238:1278 the unglycosylated E. coli-expressed BfrA (E.BfrA) 1238:1287 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 3 66 theme monomeric 616:624 arg1 glycoprotein 626:637 a monomeric glycoprotein 614:637 a monomeric glycoprotein 614:637 A β-fructosidase (BfrA) with high transglycosylation activity was purified from Aspergillus oryzae FS4 as a monomeric glycoprotein. 25501957 9 67 theme coli-expressed 1260:1273 arg1 the 1329:1331 the 1329:1331 the 1329:1331 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 67 theme coli-expressed 1260:1273 arg1 E.BfrA 1281:1286 E.BfrA 1281:1286 E.BfrA 1281:1286 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 67 theme coli-expressed 1260:1273 arg1 P. 1333:1334 P. 1333:1334 P. 1333:1334 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 9 67 theme coli-expressed 1260:1273 arg1 BfrA 1275:1278 the unglycosylated E. coli-expressed BfrA 1238:1278 the unglycosylated E. coli-expressed BfrA (E.BfrA) 1238:1287 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 10 68 theme enzyme 1760:1765 arg1 stability 1775:1783 enzyme thermal stability 1760:1783 enzyme thermal stability 1760:1783 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 5 69 theme unique 771:776 arg1 property 798:805 unique transfructosylating property 771:805 unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS 771:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 0 70 theme novel 75:79 arg1 β-fructosidase 81:94 a novel β-fructosidase 73:94 a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 73:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 7 71 theme recombinant 1000:1010 arg1 proteins 1012:1019 Both native and recombinant proteins 984:1019 Both native and recombinant proteins 984:1019 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 11 72 with fructosidase 1868:1879 arg1 capability 1913:1922 unique transfructosylating capability 1886:1922 unique transfructosylating capability of synthesizing levan- and neolevan-type FOS 1886:1967 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 7 73 theme native 989:994 arg1 proteins 1012:1019 Both native and recombinant proteins 984:1019 Both native and recombinant proteins 984:1019 Both native and recombinant proteins showed transfructosylation and hydrolyzation activities with broad substrate specificity. 25501957 1 74 from hydrolysis 254:263 arg1 substrates 306:315 various substrates 298:315 various substrates 298:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 25501957 9 75 with temperatures 1460:1471 arg1 activity 1499:1506 a maximum activity 1489:1506 a maximum activity at 55°C. Using sucrose as substrate 1489:1542 Compared with the unglycosylated E. coli-expressed BfrA (E.BfrA), the N-glycosylated native (N.BfrA) and the P. pastoris-expressed BfrA (P.BfrA) were highly stable at a wide pH range (pH 4 to 11), and significantly more thermostable at temperatures up to 50°C with a maximum activity at 55°C. Using sucrose as substrate, the Km and kcat values for total activity were 37.19±5.28 mM and 1.0016±0.039×104 s-1 for N.BfrA. 25501957 0 76 theme oryzae 113:118 arg1 FS4 120:122 Aspergillus oryzae FS4 101:122 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 5 77 theme transfructosylating 778:796 arg1 property 798:805 unique transfructosylating property 771:805 unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS 771:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 10 78 theme optima 1789:1794 arg1 activity 1796:1803 optima activity 1789:1803 optima activity 1789:1803 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 2 79 theme sucrose 420:426 arg1 concentrations 402:415 high concentrations 397:415 high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry 397:505 These enzymes also exhibit transglycosylation activity when they function with high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry. 25501957 5 80 theme neolevan-type 834:846 arg1 FOS 863:865 neolevan-type β-(2-6)-linked FOS 834:865 neolevan-type β-(2-6)-linked FOS 834:865 fructosidases that synthesize inulin-type β-(2-1)-linked FOS, BfrA has unique transfructosylating property of synthesizing levan- and neolevan-type β-(2-6)-linked FOS. 25501957 0 81 theme Aspergillus 101:111 arg1 oryzae 113:118 Aspergillus oryzae 101:118 Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 101:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 0 82 theme N-glycosylation 45:59 arg1 analysis 61:68 N-glycosylation analysis 45:68 N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 45:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 10 83 gly N-glycosylation 1671:1685 arg2 sites 1687:1691 10 of 13 putative N-glycosylation sites 1653:1691 10 of 13 putative N-glycosylation sites 1653:1691 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 10 83 gly N-glycosylation 1671:1685 arg2 13 1659:1660 13 1659:1660 13 1659:1660 Moreover, 10 of 13 putative N-glycosylation sites were glycosylated on N.BfrA, and N-glycosylation was essential for enzyme thermal stability and optima activity. 25501957 8 84 theme Glc 1166:1168 arg1 α-1 1210:1212 Glc α-1 1206:1212 Glc α-1 1206:1212 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 84 theme Glc 1166:1168 arg1 α-1 1170:1172 Glc α-1 1166:1172 Glc α-1 1166:1172 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 84 theme Glc 1166:1168 arg1 α-1 1188:1190 Glc α-1 1184:1190 Glc α-1 1184:1190 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 84 theme Glc 1166:1168 arg1 Fru 1197:1199 Fru 1197:1199 Fru 1197:1199 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 84 theme Glc 1166:1168 arg1 Fru 1219:1221 Fru 1219:1221 Fru 1219:1221 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 8 84 theme Glc 1166:1168 arg1 Fru 1179:1181 2-β Fru 1175:1181 2-β Fru 1175:1181 These proteins could hydrolyze the following linkages: Glc α-1, 2-β Fru; Glc α-1, 3-α Fru; and Glc α-1, 5-β Fru. 25501957 2 85 theme food 493:496 arg1 industry 498:505 the food industry 489:505 the food industry 489:505 These enzymes also exhibit transglycosylation activity when they function with high concentrations of sucrose, which is used to synthesize fructooligosaccharides (FOS) in the food industry. 25501957 0 86 gly N-glycosylation 45:59 arg1 β-fructosidase 81:94 a novel β-fructosidase 73:94 a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides 73:183 Purification, cloning, characterization, and N-glycosylation analysis of a novel β-fructosidase from Aspergillus oryzae FS4 synthesizing levan- and neolevan-type fructooligosaccharides. 25501957 11 87 dep A. 1858:1859 arg1 oryzae 1861:1866 oryzae 1861:1866 oryzae 1861:1866 Thus, BfrA has demonstrated as a well-characterized A. oryzae fructosidase with unique transfructosylating capability of synthesizing levan- and neolevan-type FOS. 25501957 1 88 theme terminal 268:275 arg1 units 287:291 terminal fructosyl units 268:291 terminal fructosyl units from various substrates 268:315 β-Fructosidases are a widespread group of enzymes that catalyze the hydrolysis of terminal fructosyl units from various substrates. 29185761 0 0 theme Backbone 73:80 arg1 Structure 82:90 Its Backbone Structure 69:90 Its Backbone Structure 69:90 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 2 1 theme backbone 345:352 arg1 assembly 307:314 assembly 307:314 assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step 307:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 2 2 theme hexasaccharide 330:343 arg1 backbone 345:352 the linear hexasaccharide backbone 319:352 the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step 319:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 1 3 theme nonasaccharide 232:245 arg1 KH-1 262:265 the nonasaccharide cancer antigen KH-1 228:265 the nonasaccharide cancer antigen KH-1 228:265 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 2 4 from backbone 435:442 arg1 step 451:454 one step 447:454 one step 447:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 1 5 theme cancer 247:252 arg1 KH-1 262:265 the nonasaccharide cancer antigen KH-1 228:265 the nonasaccharide cancer antigen KH-1 228:265 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 0 6 theme Structure 82:90 arg1 Assembly 57:64 Block Assembly 51:64 Block Assembly of Its Backbone Structure 51:90 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 1 7 theme antigen 254:260 arg1 KH-1 262:265 the nonasaccharide cancer antigen KH-1 228:265 the nonasaccharide cancer antigen KH-1 228:265 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 0 8 theme One-Step 104:111 arg1 Grafting 113:120 One-Step Grafting 104:120 One-Step Grafting of Three Fucose Residues 104:145 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 1 9 theme convergent 158:167 arg1 strategy 184:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 1 10 theme KH-1 262:265 arg1 synthesis 215:223 the synthesis 211:223 the synthesis of the nonasaccharide cancer antigen KH-1 211:265 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 0 11 theme Cancer-Associated 17:33 arg1 Antigen 40:46 the Cancer-Associated KH-1 Antigen 13:46 the Cancer-Associated KH-1 Antigen 13:46 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 2 12 theme residues 417:424 arg1 grafting 392:399 grafting 392:399 grafting of three fucose residues onto the backbone in one step 392:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 1 13 theme robust 150:155 arg1 strategy 184:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 29185761 2 14 theme fucose 410:415 arg1 residues 417:424 three fucose residues 404:424 three fucose residues onto the backbone in one step 404:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 0 15 theme Antigen 40:46 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure 0:90 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 0 16 theme Fucose 131:136 arg1 Residues 138:145 Three Fucose Residues 125:145 Three Fucose Residues 125:145 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 0 17 theme KH-1 35:38 arg1 Antigen 40:46 the Cancer-Associated KH-1 Antigen 13:46 the Cancer-Associated KH-1 Antigen 13:46 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 0 18 theme Block 51:55 arg1 Assembly 57:64 Block Assembly 51:64 Block Assembly of Its Backbone Structure 51:90 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 2 19 theme linear 323:328 arg1 backbone 345:352 the linear hexasaccharide backbone 319:352 the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step 319:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 0 20 theme Residues 138:145 arg1 Grafting 113:120 One-Step Grafting 104:120 One-Step Grafting of Three Fucose Residues 104:145 Synthesis of the Cancer-Associated KH-1 Antigen by Block Assembly of Its Backbone Structure Followed by One-Step Grafting of Three Fucose Residues. 29185761 2 21 dep featured 282:289 arg1 block 301:305 block 301:305 featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step 282:454 This strategy featured a one-pot block assembly of the linear hexasaccharide backbone using three disaccharides followed by grafting of three fucose residues onto the backbone in one step. 29185761 1 22 theme efficient 174:182 arg1 strategy 184:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy 148:191 A robust, convergent, and efficient strategy was developed for the synthesis of the nonasaccharide cancer antigen KH-1. 25193875 5 0 located located 703:709 arg1 hemolymph 731:739 hemolymph 731:739 hemolymph 731:739 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 5 0 located located 703:709 arg1 body 722:725 the fat body 714:725 the fat body 714:725 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 5 0 located located 703:709 arg2 BmGlcNAcase2 678:689 BmGlcNAcase2 678:689 BmGlcNAcase2 678:689 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 5 0 located located 703:709 arg2 BmFDL 668:672 BmFDL 668:672 BmFDL 668:672 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 8 1 theme N-glycan 1185:1192 arg1 levels 1163:1168 modest levels 1156:1168 modest levels of GlcNAc-type N-glycan 1156:1192 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 6 2 theme many 842:845 arg1 glycoproteins 847:859 many glycoproteins 842:859 many glycoproteins 842:859 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 6 3 theme fat 760:762 arg1 body 764:767 The fat body 756:767 The fat body 756:767 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 6 3 theme fat 760:762 arg1 tissue 781:786 the main tissue 772:786 the main tissue of recombinant protein expression by baculovirus 772:835 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 7 4 theme N-glycan 985:992 arg1 levels 994:999 GlcNAc-type N-glycan levels 973:999 GlcNAc-type N-glycan levels 973:999 These results suggest that inhibition of BmFDL and BmGlcNAcase2 could increase GlcNAc-type N-glycan levels. 25193875 8 5 theme GlcNAc-type 1173:1183 arg1 N-glycan 1185:1192 GlcNAc-type N-glycan 1173:1192 GlcNAc-type N-glycan 1173:1192 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 1 6 theme high-level 197:206 arg1 expression 208:217 high-level expression 197:217 high-level expression 197:217 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 2 7 attach presence 296:303 arg1 glycoproteins 341:353 glycoproteins 341:353 glycoproteins 341:353 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 2 7 attach presence 296:303 arg2 N-glycan 329:336 paucimannosidic-type N-glycan 308:336 paucimannosidic-type N-glycan 308:336 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 8 8 dep observed 1199:1206 arg1 % 1212:1212 0.8% 1209:1212 0.8% of total N-glycan 1209:1230 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 8 8 dep observed 1199:1206 arg1 N-glycan 1223:1230 total N-glycan 1217:1230 total N-glycan 1217:1230 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 8 9 theme glycoprotein 1117:1128 arg1 structure 1100:1108 the N-glycan structure 1087:1108 the N-glycan structure of the glycoprotein expressed by baculovirus 1087:1153 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 2 10 theme clinical 371:378 arg1 use 380:382 their clinical use 365:382 their clinical use 365:382 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 0 11 from β-N-acetylglucosaminidases 57:82 arg1 silkworm 87:94 silkworm 87:94 silkworm 87:94 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 11 12 from GlcNAcases 1592:1601 arg1 silkworm 1606:1613 silkworm 1606:1613 silkworm 1606:1613 We conclude that the structure of N-glycan can be changed by inhibiting the GlcNAcases in silkworm. 25193875 6 13 theme recombinant 791:801 arg1 expression 811:820 recombinant protein expression 791:820 recombinant protein expression 791:820 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 3 14 theme membrane-binding-type 446:466 arg1 β-N-acetylglucosaminidase 468:492 insect-specific membrane-binding-type β-N-acetylglucosaminidase 430:492 insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase) 430:504 Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). 25193875 3 14 theme membrane-binding-type 446:466 arg1 GlcNAcase 495:503 GlcNAcase 495:503 GlcNAcase 495:503 Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). 25193875 8 15 from changes 1076:1082 arg1 structure 1100:1108 the N-glycan structure 1087:1108 the N-glycan structure of the glycoprotein expressed by baculovirus 1087:1153 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 0 16 from suppression 42:52 arg1 silkworm 87:94 silkworm 87:94 silkworm 87:94 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 8 17 theme modest 1156:1161 arg1 levels 1163:1168 modest levels 1156:1168 modest levels of GlcNAc-type N-glycan 1156:1192 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 2 18 from presence 296:303 arg1 glycoproteins 341:353 glycoproteins 341:353 glycoproteins 341:353 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 2 19 gly glycoproteins 341:353 arg1 glycoproteins 341:353 glycoproteins 341:353 glycoproteins 341:353 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 8 20 gly glycoprotein 1117:1128 arg1 glycoprotein 1117:1128 the glycoprotein 1113:1128 the glycoprotein expressed by baculovirus 1113:1153 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 8 21 theme GlcNAcase 1026:1034 arg1 inhibitor 1036:1044 a GlcNAcase inhibitor 1024:1044 a GlcNAcase inhibitor 1024:1044 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 9 22 theme transcript 1326:1335 arg1 level 1337:1341 the BmFDL transcript level 1316:1341 the BmFDL transcript level 1316:1341 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 1 23 theme baculovirus-silkworm 101:120 arg1 system 153:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 23 theme baculovirus-silkworm 101:120 arg1 method 176:181 an excellent method 163:181 an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation 163:280 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 24 theme post-translational 223:240 arg1 modifications 242:254 post-translational modifications 223:254 post-translational modifications 223:254 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 7 25 theme BmFDL 935:939 arg1 inhibition 921:930 inhibition 921:930 inhibition of BmFDL and BmGlcNAcase2 921:956 These results suggest that inhibition of BmFDL and BmGlcNAcase2 could increase GlcNAc-type N-glycan levels. 25193875 5 26 theme fat 718:720 arg1 body 722:725 the fat body 714:725 the fat body 714:725 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 1 27 theme recombinant 122:132 arg1 system 153:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 27 theme recombinant 122:132 arg1 method 176:181 an excellent method 163:181 an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation 163:280 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 6 28 theme expression 811:820 arg1 body 764:767 The fat body 756:767 The fat body 756:767 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 6 28 theme expression 811:820 arg1 tissue 781:786 the main tissue 772:786 the main tissue of recombinant protein expression by baculovirus 772:835 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 0 29 theme structure 29:37 arg1 Improvement 0:10 Improvement 0:10 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 0:95 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 10 30 theme RNAi-transgenic 1490:1504 arg1 silkworm 1506:1513 the RNAi-transgenic silkworm 1486:1513 the RNAi-transgenic silkworm 1486:1513 The proportion of GlcNAc-type N-glycan increased to 4.3% in the RNAi-transgenic silkworm. 25193875 8 31 theme N-glycan 1223:1230 arg1 % 1212:1212 0.8% 1209:1212 0.8% of total N-glycan 1209:1230 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 8 31 theme N-glycan 1223:1230 arg1 N-glycan 1223:1230 total N-glycan 1217:1230 total N-glycan 1217:1230 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 1 32 theme protein 134:140 arg1 system 153:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 32 theme protein 134:140 arg1 method 176:181 an excellent method 163:181 an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation 163:280 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 6 33 theme protein 803:809 arg1 expression 811:820 recombinant protein expression 791:820 recombinant protein expression 791:820 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 0 34 theme glycosylation 15:27 arg1 structure 29:37 glycosylation structure 15:37 glycosylation structure 15:37 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 4 35 from GlcNAcases 588:597 arg1 silkworm 514:521 the silkworm 510:521 the silkworm 510:521 In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. 25193875 4 36 theme membrane-binding-type 566:586 arg1 BmGlcNAcase1 524:535 BmGlcNAcase1 524:535 BmGlcNAcase1 524:535 In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. 25193875 4 36 theme membrane-binding-type 566:586 arg1 BmGlcNAcase2 538:549 BmGlcNAcase2 538:549 BmGlcNAcase2 538:549 In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. 25193875 4 36 theme membrane-binding-type 566:586 arg1 BmFDL 556:560 BmFDL 556:560 BmFDL 556:560 In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. 25193875 4 36 theme membrane-binding-type 566:586 arg1 GlcNAcases 588:597 membrane-binding-type GlcNAcases 566:597 membrane-binding-type GlcNAcases 566:597 In the silkworm, BmGlcNAcase1, BmGlcNAcase2, and BmFDL are membrane-binding-type GlcNAcases. 25193875 1 37 theme expression 142:151 arg1 system 153:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 37 theme expression 142:151 arg1 method 176:181 an excellent method 163:181 an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation 163:280 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 5 38 from hemolymph 731:739 arg1 located 703:709 located 703:709 located 703:709 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 5 39 from located 703:709 arg1 hemolymph 731:739 hemolymph 731:739 hemolymph 731:739 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 5 39 from located 703:709 arg1 body 722:725 the fat body 714:725 the fat body 714:725 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 8 40 theme total 1217:1221 arg1 N-glycan 1223:1230 total N-glycan 1217:1230 total N-glycan 1217:1230 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 0 41 from silkworm 87:94 arg1 suppression 42:52 suppression 42:52 suppression of β-N-acetylglucosaminidases in silkworm 42:94 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 9 42 theme RNA 1284:1286 arg1 RNAi 1302:1305 RNAi 1302:1305 RNAi 1302:1305 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 9 42 theme RNA 1284:1286 arg1 interference 1288:1299 RNA interference 1284:1299 RNA interference (RNAi) 1284:1306 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 0 43 theme β-N-acetylglucosaminidases 57:82 arg1 suppression 42:52 suppression 42:52 suppression of β-N-acetylglucosaminidases in silkworm 42:94 Improvement of glycosylation structure by suppression of β-N-acetylglucosaminidases in silkworm. 25193875 7 44 theme GlcNAc-type 973:983 arg1 levels 994:999 GlcNAc-type N-glycan levels 973:999 GlcNAc-type N-glycan levels 973:999 These results suggest that inhibition of BmFDL and BmGlcNAcase2 could increase GlcNAc-type N-glycan levels. 25193875 3 45 theme Paucimannosidic-type 385:404 arg1 N-glycan 406:413 Paucimannosidic-type N-glycan 385:413 Paucimannosidic-type N-glycan 385:413 Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). 25193875 9 46 theme transgenic 1255:1264 arg1 silkworm 1266:1273 a transgenic silkworm 1253:1273 a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm 1253:1423 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 5 47 from body 722:725 arg1 located 703:709 located 703:709 located 703:709 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 9 48 theme BmFDL 1320:1324 arg1 level 1337:1341 the BmFDL transcript level 1316:1341 the BmFDL transcript level 1316:1341 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 6 49 theme main 776:779 arg1 body 764:767 The fat body 756:767 The fat body 756:767 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 6 49 theme main 776:779 arg1 tissue 781:786 the main tissue 772:786 the main tissue of recombinant protein expression by baculovirus 772:835 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 25193875 2 50 theme N-glycan 329:336 arg1 presence 296:303 the presence 292:303 the presence of paucimannosidic-type N-glycan in glycoproteins 292:353 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 1 51 theme excellent 166:174 arg1 system 153:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system 97:158 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 1 51 theme excellent 166:174 arg1 method 176:181 an excellent method 163:181 an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation 163:280 The baculovirus-silkworm recombinant protein expression system is an excellent method for achieving high-level expression and post-translational modifications, especially glycosylation. 25193875 7 52 theme BmGlcNAcase2 945:956 arg1 inhibition 921:930 inhibition 921:930 inhibition of BmFDL and BmGlcNAcase2 921:956 These results suggest that inhibition of BmFDL and BmGlcNAcase2 could increase GlcNAc-type N-glycan levels. 25193875 5 53 theme GlcNAcases 642:651 arg1 localization 620:631 the localization 616:631 the localization of these GlcNAcases 616:651 We investigated the localization of these GlcNAcases and found that BmFDL and BmGlcNAcase2 were mainly located in the fat body and hemolymph, respectively. 25193875 10 54 theme N-glycan 1456:1463 arg1 proportion 1430:1439 The proportion 1426:1439 The proportion of GlcNAc-type N-glycan 1426:1463 The proportion of GlcNAc-type N-glycan increased to 4.3% in the RNAi-transgenic silkworm. 25193875 2 55 theme paucimannosidic-type 308:327 arg1 N-glycan 329:336 paucimannosidic-type N-glycan 308:336 paucimannosidic-type N-glycan 308:336 However, the presence of paucimannosidic-type N-glycan in glycoproteins restricts their clinical use. 25193875 8 56 theme N-glycan 1091:1098 arg1 structure 1100:1108 the N-glycan structure 1087:1108 the N-glycan structure of the glycoprotein expressed by baculovirus 1087:1153 We therefore injected a GlcNAcase inhibitor into silkworms to investigate changes in the N-glycan structure of the glycoprotein expressed by baculovirus; modest levels of GlcNAc-type N-glycan were observed (0.8% of total N-glycan). 25193875 11 57 theme N-glycan 1550:1557 arg1 structure 1537:1545 the structure 1533:1545 the structure of N-glycan 1533:1557 We conclude that the structure of N-glycan can be changed by inhibiting the GlcNAcases in silkworm. 25193875 3 58 theme insect-specific 430:444 arg1 β-N-acetylglucosaminidase 468:492 insect-specific membrane-binding-type β-N-acetylglucosaminidase 430:492 insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase) 430:504 Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). 25193875 3 58 theme insect-specific 430:444 arg1 GlcNAcase 495:503 GlcNAcase 495:503 GlcNAcase 495:503 Paucimannosidic-type N-glycan is produced by insect-specific membrane-binding-type β-N-acetylglucosaminidase (GlcNAcase). 25193875 9 59 theme enzyme 1347:1352 arg1 activity 1354:1361 enzyme activity 1347:1361 enzyme activity 1347:1361 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 10 60 theme GlcNAc-type 1444:1454 arg1 N-glycan 1456:1463 GlcNAc-type N-glycan 1444:1463 GlcNAc-type N-glycan 1444:1463 The proportion of GlcNAc-type N-glycan increased to 4.3% in the RNAi-transgenic silkworm. 25193875 9 61 theme control 1408:1414 arg1 silkworm 1416:1423 the control silkworm 1404:1423 the control silkworm 1404:1423 Next, we generated a transgenic silkworm in which RNA interference (RNAi) reduced the BmFDL transcript level and enzyme activity to 25% and 50%, respectively, of that of the control silkworm. 25193875 6 62 gly glycoproteins 847:859 arg1 glycoproteins 847:859 many glycoproteins 842:859 many glycoproteins 842:859 The fat body is the main tissue of recombinant protein expression by baculovirus, and many glycoproteins are secreted into the hemolymph. 27118590 7 0 theme protective 1190:1199 arg1 response 1210:1217 a potentially protective antibody response 1176:1217 a potentially protective antibody response 1176:1217 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 13 1 theme protein 2080:2086 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 15 2 theme glycosylation 2533:2545 arg1 potential 2503:2511 the application potential 2487:2511 the application potential of O-linked protein glycosylation 2487:2545 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 9 3 theme substrate 1485:1493 arg1 glycosylation 1495:1507 substrate glycosylation 1485:1507 substrate glycosylation 1485:1507 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 6 4 theme polysaccharide 942:955 arg1 structure 957:965 structure 957:965 structure 957:965 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 10 5 theme simpler 1657:1663 arg1 strategy 1681:1688 a simpler and more robust strategy 1655:1688 a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens 1655:1755 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 15 6 theme different 2642:2650 arg1 pathogens 2652:2660 different pathogens 2642:2660 different pathogens 2642:2660 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 5 7 theme vector 681:686 arg1 construction 651:662 the construction 647:662 the construction of an expression vector for polysaccharide synthesis 647:715 We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. 27118590 6 8 theme glycosylation 829:841 arg1 system 843:848 the O-linked glycosylation system 816:848 the O-linked glycosylation system 816:848 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 4 9 dep system 450:455 arg1 glycosylation 467:479 O-linked glycosylation 458:479 O-linked glycosylation via Neisseria meningitidis 458:506 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 14 10 theme next-generation 2330:2344 arg1 vaccines 2359:2366 next-generation bioconjugate vaccines 2330:2366 next-generation bioconjugate vaccines 2330:2366 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 2 11 theme N-linked 266:273 arg1 systems 289:295 N-linked glycosylation systems 266:295 N-linked glycosylation systems 266:295 Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. 27118590 7 12 theme O-linked 1138:1145 arg1 system 1161:1166 this O-linked glycosylation system 1133:1166 this O-linked glycosylation system 1133:1166 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 9 13 theme optimized 1531:1539 arg1 system 1541:1546 this optimized system 1526:1546 this optimized system 1526:1546 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 6 14 theme glycosylation 1019:1031 arg1 system 1033:1038 the N-linked glycosylation system 1006:1038 the N-linked glycosylation system 1006:1038 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 16 15 theme subunit 2840:2846 arg1 candidates 2856:2865 protein subunit vaccine candidates 2832:2865 protein subunit vaccine candidates 2832:2865 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 13 16 theme effective 2264:2272 arg1 vaccines 2284:2291 effective conjugate vaccines 2264:2291 effective conjugate vaccines 2264:2291 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 16 17 attach attached 2723:2730 arg3 future 2670:2675 future 2670:2675 future 2670:2675 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 16 17 attach attached 2723:2730 arg1 complex 2760:2766 major histocompatibility complex 2735:2766 major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation 2735:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 16 17 attach attached 2723:2730 arg2 polysaccharides 2698:2712 bacterial antigenic polysaccharides 2678:2712 bacterial antigenic polysaccharides 2678:2712 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 4 18 theme O-linked 458:465 arg1 glycosylation 467:479 O-linked glycosylation 458:479 O-linked glycosylation via Neisseria meningitidis 458:506 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 5 19 theme polysaccharide 692:705 arg1 synthesis 707:715 polysaccharide synthesis 692:715 polysaccharide synthesis 692:715 We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. 27118590 9 20 theme future 1567:1572 arg1 development 1582:1592 future vaccine development 1567:1592 future vaccine development 1567:1592 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 11 21 link N-linked 1861:1868 arg1 glycosylation 1878:1890 N-linked protein glycosylation 1861:1890 N-linked protein glycosylation 1861:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 13 22 theme attenuated 2232:2241 arg1 pathogens 2243:2251 attenuated pathogens 2232:2251 attenuated pathogens 2232:2251 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 4 23 link O-linked 458:465 arg1 glycosylation 467:479 O-linked glycosylation 458:479 O-linked glycosylation via Neisseria meningitidis 458:506 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 4 24 theme protein 428:434 arg1 system 450:455 a novel protein glycosylation system 420:455 a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine 420:577 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 11 25 theme bioconjugate 1834:1845 arg1 vaccines 1847:1854 bioconjugate vaccines 1834:1854 bioconjugate vaccines with N-linked protein glycosylation 1834:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 15 26 theme bioconjugate 2612:2623 arg1 vaccines 2625:2632 bioconjugate vaccines 2612:2632 bioconjugate vaccines against different pathogens 2612:2660 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 1 27 theme effective 137:145 arg1 types 158:162 the most effective and safest types 128:162 the most effective and safest types of vaccines against bacterial pathogens 128:202 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 14 28 theme vaccines 2359:2366 arg1 design 2320:2325 the further design 2308:2325 the further design of next-generation bioconjugate vaccines 2308:2366 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 15 29 theme robust 2582:2587 arg1 strategy 2589:2596 a simpler and more robust strategy 2563:2596 a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens 2563:2660 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 15 30 theme application 2491:2501 arg1 potential 2503:2511 the application potential 2487:2511 the application potential of O-linked protein glycosylation 2487:2545 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 1 31 theme safest 151:156 arg1 types 158:162 the most effective and safest types 128:162 the most effective and safest types of vaccines against bacterial pathogens 128:202 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 11 32 theme protein 1870:1876 arg1 glycosylation 1878:1890 N-linked protein glycosylation 1861:1890 N-linked protein glycosylation 1861:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 5 33 theme Shigella 623:630 arg1 spp. 632:635 Shigella spp. 623:635 Shigella spp. 623:635 We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. 27118590 6 34 gly glycosylated 776:787 arg1 substrates 758:767 different protein substrates 740:767 different protein substrates 740:767 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 0 35 theme Glycosylation 53:65 arg1 System 67:72 an O-Linked Glycosylation System 41:72 an O-Linked Glycosylation System 41:72 Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System. 27118590 11 36 gly glycosylation 1878:1890 arg1 vaccines 1847:1854 bioconjugate vaccines 1834:1854 bioconjugate vaccines with N-linked protein glycosylation 1834:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 10 37 theme pathogens 1747:1755 arg1 pathogens 1747:1755 pathogens 1747:1755 pathogens 1747:1755 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 10 37 theme pathogens 1747:1755 arg1 variety 1736:1742 a variety 1734:1742 a variety of pathogens 1734:1755 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 9 38 theme peptides 1382:1389 arg1 fusion 1366:1371 the fusion 1362:1371 the fusion of other peptides recognized by major histocompatibility complex class II around MOOR 1362:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 16 39 theme immunological 2796:2808 arg1 memory 2810:2815 immunological memory 2796:2815 immunological memory 2796:2815 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 1 40 theme Conjugate 86:94 arg1 vaccines 96:103 UNLABELLED Conjugate vaccines 75:103 UNLABELLED Conjugate vaccines 75:103 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 14 41 theme 8 amino 2416:2422 arg1 acids 2424:2428 8 amino acids 2416:2428 8 amino acids 2416:2428 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 9 42 contain had 1459:1461 arg1 fusion 1366:1371 the fusion 1362:1371 the fusion of other peptides recognized by major histocompatibility complex class II around MOOR 1362:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 9 42 contain had 1459:1461 arg2 effects 1474:1480 no adverse effects 1463:1480 no adverse effects 1463:1480 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 0 43 theme Conjugate 16:24 arg1 Vaccines 26:33 Conjugate Vaccines 16:33 Conjugate Vaccines 16:33 Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System. 27118590 9 44 theme histocompatibility 1411:1428 arg1 class 1438:1442 major histocompatibility complex class II 1405:1445 major histocompatibility complex class II around MOOR 1405:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 12 45 theme synthetic 1984:1992 arg1 pathway 1994:2000 the exogenous polysaccharide synthetic pathway 1955:2000 the exogenous polysaccharide synthetic pathway in Escherichia coli 1955:2020 However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. 27118590 14 46 theme short 2390:2394 arg1 sufficient 2438:2447 sufficient 2438:2447 sufficient 2438:2447 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 14 46 theme short 2390:2394 arg1 motif 2396:2400 a novel short motif 2382:2400 a novel short motif consisting of 8 amino acids that is sufficient for glycosylation 2382:2465 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 6 47 theme Gram-negative 889:901 arg1 strains 928:934 some strains 923:934 some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system 923:1038 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 6 47 theme Gram-negative 889:901 arg1 bacteria 903:910 other Gram-negative bacteria 883:910 other Gram-negative bacteria 883:910 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 11 48 theme rapid 1783:1787 arg1 development 1789:1799 the rapid development 1779:1799 the rapid development of synthetic biology 1779:1820 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 0 49 theme O-Linked 44:51 arg1 System 67:72 an O-Linked Glycosylation System 41:72 an O-Linked Glycosylation System 41:72 Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System. 27118590 11 50 dep Recently 1769:1776 arg1 IMPORTANCE 1758:1767 IMPORTANCE 1758:1767 IMPORTANCE 1758:1767 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 16 51 theme antigenic 2688:2696 arg1 polysaccharides 2698:2712 bacterial antigenic polysaccharides 2678:2712 bacterial antigenic polysaccharides 2678:2712 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 13 52 theme structure 2156:2164 arg1 specificity 2166:2176 a lower structure specificity 2148:2176 a lower structure specificity for sugar substrates 2148:2197 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 12 53 theme exogenous 1959:1967 arg1 pathway 1994:2000 the exogenous polysaccharide synthetic pathway 1955:2000 the exogenous polysaccharide synthetic pathway in Escherichia coli 1955:2020 However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. 27118590 14 54 theme novel 2384:2388 arg1 sufficient 2438:2447 sufficient 2438:2447 sufficient 2438:2447 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 14 54 theme novel 2384:2388 arg1 motif 2396:2400 a novel short motif 2382:2400 a novel short motif consisting of 8 amino acids that is sufficient for glycosylation 2382:2465 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 11 55 theme biology 1814:1820 arg1 development 1789:1799 the rapid development 1779:1799 the rapid development of synthetic biology 1779:1820 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 9 56 theme adverse 1466:1472 arg1 effects 1474:1480 no adverse effects 1463:1480 no adverse effects 1463:1480 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 6 57 link O-linked 820:827 arg1 system 843:848 the O-linked glycosylation system 816:848 the O-linked glycosylation system 816:848 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 16 58 theme histocompatibility 2741:2758 arg1 complex 2760:2766 major histocompatibility complex 2735:2766 major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation 2735:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 16 59 theme immune 2885:2890 arg1 stimulation 2892:2902 double immune stimulation 2878:2902 double immune stimulation 2878:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 13 60 contain has 2144:2146 arg2 specificity 2166:2176 a lower structure specificity 2148:2176 a lower structure specificity for sugar substrates 2148:2197 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 13 60 contain has 2144:2146 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 10 61 theme glycoengineering 1618:1633 arg1 toolbox 1635:1641 the glycoengineering toolbox 1614:1641 the glycoengineering toolbox 1614:1641 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 6 62 theme different 740:748 arg1 substrates 758:767 different protein substrates 740:767 different protein substrates 740:767 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 16 63 theme bacterial 2678:2686 arg1 polysaccharides 2698:2712 bacterial antigenic polysaccharides 2678:2712 bacterial antigenic polysaccharides 2678:2712 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 5 64 theme expression 670:679 arg1 vector 681:686 an expression vector 667:686 an expression vector for polysaccharide synthesis 667:715 We successfully established this system in Shigella spp., avoiding the construction of an expression vector for polysaccharide synthesis. 27118590 13 65 theme glycosylation 2088:2100 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 14 66 theme bioconjugate 2346:2357 arg1 vaccines 2359:2366 next-generation bioconjugate vaccines 2330:2366 next-generation bioconjugate vaccines 2330:2366 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 7 67 theme glycosylation 1147:1159 arg1 system 1161:1166 this O-linked glycosylation system 1133:1166 this O-linked glycosylation system 1133:1166 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 6 68 theme O-linked 820:827 arg1 system 843:848 the O-linked glycosylation system 816:848 the O-linked glycosylation system 816:848 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 2 69 theme glycosylation 275:287 arg1 systems 289:295 N-linked glycosylation systems 266:295 N-linked glycosylation systems 266:295 Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. 27118590 1 70 theme types 158:162 arg1 types 158:162 the most effective and safest types 128:162 the most effective and safest types of vaccines against bacterial pathogens 128:202 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 1 70 theme types 158:162 arg1 one 121:123 one 121:123 one 121:123 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 1 71 theme vaccines 167:174 arg1 types 158:162 the most effective and safest types 128:162 the most effective and safest types of vaccines against bacterial pathogens 128:202 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 8 72 theme O-glycosyltransferase 1304:1324 arg1 PglL 1326:1329 the O-glycosyltransferase PglL 1300:1329 the O-glycosyltransferase PglL 1300:1329 Furthermore, we elucidated and optimized the recognition motif, named MOOR, for the O-glycosyltransferase PglL. 27118590 10 73 theme robust 1674:1679 arg1 strategy 1681:1688 a simpler and more robust strategy 1655:1688 a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens 1655:1755 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 2 74 link N-linked 266:273 arg1 systems 289:295 N-linked glycosylation systems 266:295 N-linked glycosylation systems 266:295 Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. 27118590 13 75 theme O-linked 2071:2078 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 1 76 theme bacterial 184:192 arg1 pathogens 194:202 bacterial pathogens 184:202 bacterial pathogens 184:202 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 8 77 theme recognition 1265:1275 arg1 motif 1277:1281 the recognition motif 1261:1281 the recognition motif 1261:1281 Furthermore, we elucidated and optimized the recognition motif, named MOOR, for the O-glycosyltransferase PglL. 27118590 6 78 theme N-linked 1010:1017 arg1 system 1033:1038 the N-linked glycosylation system 1006:1038 the N-linked glycosylation system 1006:1038 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 7 79 from series 1060:1065 arg1 results 1045:1051 The results 1041:1051 The results from a series of animal experiments 1041:1087 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 7 80 theme conjugate 1103:1111 arg1 vaccine 1113:1119 the conjugate vaccine 1099:1119 the conjugate vaccine produced by this O-linked glycosylation system 1099:1166 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 6 81 link N-linked 1010:1017 arg1 system 1033:1038 the N-linked glycosylation system 1006:1038 the N-linked glycosylation system 1006:1038 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 13 82 from meningitidis 2124:2135 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 16 83 theme protein 2832:2838 arg1 candidates 2856:2865 protein subunit vaccine candidates 2832:2865 protein subunit vaccine candidates 2832:2865 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 14 84 theme further 2312:2318 arg1 design 2320:2325 the further design 2308:2325 the further design of next-generation bioconjugate vaccines 2308:2366 To facilitate the further design of next-generation bioconjugate vaccines, we optimized a novel short motif consisting of 8 amino acids that is sufficient for glycosylation. 27118590 10 85 theme bioconjugate 1704:1715 arg1 vaccines 1717:1724 bioconjugate vaccines 1704:1724 bioconjugate vaccines against a variety of pathogens 1704:1755 Our results expand the glycoengineering toolbox and provide a simpler and more robust strategy for producing bioconjugate vaccines against a variety of pathogens. 27118590 13 86 theme conjugate 2274:2282 arg1 vaccines 2284:2291 effective conjugate vaccines 2264:2291 effective conjugate vaccines 2264:2291 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 2 87 theme vaccine 217:223 arg1 biosynthesis 225:236 vaccine biosynthesis 217:236 vaccine biosynthesis 217:236 Previously, vaccine biosynthesis has been performed by using N-linked glycosylation systems. 27118590 7 88 theme animal 1070:1075 arg1 experiments 1077:1087 animal experiments 1070:1087 animal experiments 1070:1087 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 16 89 theme vaccine 2848:2854 arg1 candidates 2856:2865 protein subunit vaccine candidates 2832:2865 protein subunit vaccine candidates 2832:2865 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 3 90 theme structural 311:320 arg1 specificity 322:332 the structural specificity 307:332 the structural specificity of these systems for sugar substrates 307:370 However, the structural specificity of these systems for sugar substrates has hindered their application. 27118590 4 91 theme novel 422:426 arg1 system 450:455 a novel protein glycosylation system 420:455 a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine 420:577 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 7 92 theme experiments 1077:1087 arg1 series 1060:1065 a series 1058:1065 a series of animal experiments 1058:1087 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 15 93 theme O-linked 2516:2523 arg1 glycosylation 2533:2545 O-linked protein glycosylation 2516:2545 O-linked protein glycosylation 2516:2545 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 9 94 theme vaccine 1574:1580 arg1 development 1582:1592 future vaccine development 1567:1592 future vaccine development 1567:1592 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 16 95 theme major 2735:2739 arg1 complex 2760:2766 major histocompatibility complex 2735:2766 major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation 2735:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 4 96 theme glycosylation 436:448 arg1 system 450:455 a novel protein glycosylation system 420:455 a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine 420:577 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 11 97 theme N-linked 1861:1868 arg1 glycosylation 1878:1890 N-linked protein glycosylation 1861:1890 N-linked protein glycosylation 1861:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 11 98 with vaccines 1847:1854 arg1 glycosylation 1878:1890 N-linked protein glycosylation 1861:1890 N-linked protein glycosylation 1861:1890 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 15 99 link O-linked 2516:2523 arg1 glycosylation 2533:2545 O-linked protein glycosylation 2516:2545 O-linked protein glycosylation 2516:2545 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 3 100 theme systems 343:349 arg1 specificity 322:332 the structural specificity 307:332 the structural specificity of these systems for sugar substrates 307:370 However, the structural specificity of these systems for sugar substrates has hindered their application. 27118590 12 101 from pathway 1994:2000 arg1 coli 2017:2020 Escherichia coli 2005:2020 Escherichia coli 2005:2020 However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. 27118590 4 102 theme Neisseria 485:493 arg1 meningitidis 495:506 Neisseria meningitidis 485:506 Neisseria meningitidis 485:506 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 9 103 theme other 1376:1380 arg1 peptides 1382:1389 other peptides 1376:1389 other peptides recognized by major histocompatibility complex class II around MOOR 1376:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 1 104 theme UNLABELLED 75:84 arg1 vaccines 96:103 UNLABELLED Conjugate vaccines 75:103 UNLABELLED Conjugate vaccines 75:103 UNLABELLED Conjugate vaccines are known to be one of the most effective and safest types of vaccines against bacterial pathogens. 27118590 3 105 theme sugar 355:359 arg1 substrates 361:370 sugar substrates 355:370 sugar substrates 355:370 However, the structural specificity of these systems for sugar substrates has hindered their application. 27118590 7 106 link O-linked 1138:1145 arg1 system 1161:1166 this O-linked glycosylation system 1133:1166 this O-linked glycosylation system 1133:1166 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 15 107 theme simpler 2565:2571 arg1 strategy 2589:2596 a simpler and more robust strategy 2563:2596 a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens 2563:2660 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 4 108 theme conjugate 561:569 arg1 vaccine 571:577 a conjugate vaccine 559:577 a conjugate vaccine 559:577 Here, we report a novel protein glycosylation system (O-linked glycosylation via Neisseria meningitidis) that can transfer virtually any glycan to produce a conjugate vaccine. 27118590 0 109 theme Vaccines 26:33 arg1 Biosynthesis 0:11 Biosynthesis 0:11 Biosynthesis of Conjugate Vaccines 0:33 Biosynthesis of Conjugate Vaccines Using an O-Linked Glycosylation System. 27118590 9 110 theme major 1405:1409 arg1 class 1438:1442 major histocompatibility complex class II 1405:1445 major histocompatibility complex class II around MOOR 1405:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 9 111 theme complex 1430:1436 arg1 class 1438:1442 major histocompatibility complex class II 1405:1445 major histocompatibility complex class II around MOOR 1405:1457 Finally, we demonstrated that the fusion of other peptides recognized by major histocompatibility complex class II around MOOR had no adverse effects on substrate glycosylation, suggesting that this optimized system will be useful for future vaccine development. 27118590 7 112 theme antibody 1201:1208 arg1 response 1210:1217 a potentially protective antibody response 1176:1217 a potentially protective antibody response 1176:1217 The results from a series of animal experiments show that the conjugate vaccine produced by this O-linked glycosylation system offered a potentially protective antibody response. 27118590 6 113 theme other 883:887 arg1 strains 928:934 some strains 923:934 some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system 923:1038 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 6 113 theme other 883:887 arg1 bacteria 903:910 other Gram-negative bacteria 883:910 other Gram-negative bacteria 883:910 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 13 114 theme sugar 2182:2186 arg1 substrates 2188:2197 sugar substrates 2182:2197 sugar substrates 2182:2197 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 13 115 theme lower 2150:2154 arg1 specificity 2166:2176 a lower structure specificity 2148:2176 a lower structure specificity for sugar substrates 2148:2197 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 15 116 theme protein 2525:2531 arg1 glycosylation 2533:2545 O-linked protein glycosylation 2516:2545 O-linked protein glycosylation 2516:2545 Our results expand the application potential of O-linked protein glycosylation and demonstrate a simpler and more robust strategy for producing bioconjugate vaccines against different pathogens. 27118590 12 117 theme polysaccharide 1969:1982 arg1 pathway 1994:2000 the exogenous polysaccharide synthetic pathway 1955:2000 the exogenous polysaccharide synthetic pathway in Escherichia coli 1955:2020 However, the difficulty of reestablishing the exogenous polysaccharide synthetic pathway in Escherichia coli hinders their application. 27118590 16 118 theme double 2878:2883 arg1 stimulation 2892:2902 double immune stimulation 2878:2902 double immune stimulation 2878:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 11 119 theme synthetic 1804:1812 arg1 biology 1814:1820 synthetic biology 1804:1820 synthetic biology 1804:1820 IMPORTANCE Recently, the rapid development of synthetic biology has allowed bioconjugate vaccines with N-linked protein glycosylation to become a reality. 27118590 13 120 theme Neisseria 2114:2122 arg1 meningitidis 2124:2135 Neisseria meningitidis 2114:2135 Neisseria meningitidis 2114:2135 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 13 121 link O-linked 2071:2078 arg1 system 2102:2107 an O-linked protein glycosylation system 2068:2107 an O-linked protein glycosylation system 2068:2107 Here, we show that an O-linked protein glycosylation system from Neisseria meningitidis, which has a lower structure specificity for sugar substrates, could be engineered directly in attenuated pathogens to produce effective conjugate vaccines. 27118590 6 122 theme protein 750:756 arg1 substrates 758:767 different protein substrates 740:767 different protein substrates 740:767 We further found that different protein substrates can be glycosylated using this system and that the O-linked glycosylation system can also effectively function in other Gram-negative bacteria, including some strains whose polysaccharide structure was not suitable for conjugation using the N-linked glycosylation system. 27118590 16 123 dep binding 2768:2774 arg1 attached 2820:2827 attached 2820:2827 attached to protein subunit vaccine candidates to provide double immune stimulation 2820:2902 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 27118590 16 123 dep binding 2768:2774 arg1 improve 2788:2794 improve 2788:2794 to improve immunological memory 2785:2815 In the future, bacterial antigenic polysaccharides could be attached to major histocompatibility complex binding peptides to improve immunological memory or attached to protein subunit vaccine candidates to provide double immune stimulation. 25773197 0 0 theme Microbacterium 98:111 arg1 paraoxydans 113:123 Microbacterium paraoxydans 98:123 Microbacterium paraoxydans 98:123 Biotransformation of sucrose into hexyl-α-D-glucopyranoside and -polyglucosides by whole cells of Microbacterium paraoxydans. 25773197 1 1 theme enzymes 195:201 arg1 activity 172:179 the transglycosylation activity 149:179 the transglycosylation activity of cell-bound enzymes from Microbacterium 149:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 1 2 theme -polyglucosides 293:307 arg1 synthesis 251:259 the synthesis 247:259 the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor 247:341 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 5 3 theme % 882:882 arg1 yield 869:873 a total yield 861:873 a total yield of ~44 % 861:882 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 5 3 theme % 882:882 arg1 h 855:855 every 6 h 847:855 every 6 h 847:855 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 6 4 theme HG 993:994 arg1 di- 1003:1005 HG, hexyl di- 993:1005 HG, hexyl di- 993:1005 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 6 5 dep HG 993:994 arg1 hexyl 997:1001 hexyl 997:1001 hexyl 997:1001 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 5 6 theme optimized 947:955 arg1 conditions 957:966 the optimized conditions 943:966 the optimized conditions 943:966 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 4 7 theme main 624:627 arg1 product 629:635 the main product 620:635 the main product 620:635 Structural elucidation of the main product (purified by solid phase chromatography) was done by HSQC (2D NMR) which was confirmed as 1-hexyl-α-D-glucopyranoside. 25773197 0 8 theme paraoxydans 113:123 arg1 cells 89:93 whole cells 83:93 whole cells of Microbacterium paraoxydans 83:123 Biotransformation of sucrose into hexyl-α-D-glucopyranoside and -polyglucosides by whole cells of Microbacterium paraoxydans. 25773197 2 9 with h 420:420 arg1 sucrose 427:433 sucrose 427:433 sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity) 427:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 5 10 theme cells 841:845 arg1 feeding 824:830 continuous feeding 813:830 continuous feeding of whole cells 813:845 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 4 11 theme solid 650:654 arg1 chromatography 662:675 solid phase chromatography 650:675 solid phase chromatography 650:675 Structural elucidation of the main product (purified by solid phase chromatography) was done by HSQC (2D NMR) which was confirmed as 1-hexyl-α-D-glucopyranoside. 25773197 3 12 theme alkyl-glucosides 528:543 arg1 and-polyglucosides 545:562 The synthesized alkyl-glucosides and-polyglucosides 512:562 The synthesized alkyl-glucosides and-polyglucosides 512:562 The synthesized alkyl-glucosides and-polyglucosides were characterized by ESI-MS. 25773197 2 13 theme lyophilized 449:459 arg1 activity 501:508 equivalent to 8 IU α-glucosidase activity 468:508 equivalent to 8 IU α-glucosidase activity 468:508 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 13 theme lyophilized 449:459 arg1 cells 461:465 lyophilized cells 449:465 lyophilized cells (equivalent to 8 IU α-glucosidase activity) 449:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 14 with sucrose 427:433 arg1 activity 501:508 equivalent to 8 IU α-glucosidase activity 468:508 equivalent to 8 IU α-glucosidase activity 468:508 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 14 with sucrose 427:433 arg1 cells 461:465 lyophilized cells 449:465 lyophilized cells (equivalent to 8 IU α-glucosidase activity) 449:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 1 15 from activity 172:179 arg1 Microbacterium 208:221 Microbacterium 208:221 Microbacterium 208:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 6 16 theme novel 1052:1056 arg1 method 1058:1063 a novel method 1050:1063 a novel method 1050:1063 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 5 17 theme continuous 813:822 arg1 feeding 824:830 continuous feeding 813:830 continuous feeding of whole cells 813:845 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 6 18 theme CONCLUSION 969:978 arg1 Synthesis 980:988 CONCLUSION Synthesis 969:988 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides 969:1024 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 2 19 theme HG 360:361 arg1 % 375:375 14.8 % 370:375 14.8 % 370:375 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 19 theme HG 360:361 arg1 yield 363:367 Maximum HG yield 352:367 RESULTS Maximum HG yield (14.8 %) 344:376 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 20 theme water 399:403 arg1 activity 405:412 0.96 water activity 394:412 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity) 394:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 1 21 theme glycosyl 328:335 arg1 donor 337:341 a glycosyl donor 326:341 a glycosyl donor 326:341 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 1 21 theme glycosyl 328:335 arg1 sucrose 315:321 sucrose 315:321 sucrose 315:321 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 2 22 theme Maximum 352:358 arg1 % 375:375 14.8 % 370:375 14.8 % 370:375 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 22 theme Maximum 352:358 arg1 yield 363:367 Maximum HG yield 352:367 RESULTS Maximum HG yield (14.8 %) 344:376 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 4 23 theme phase 656:660 arg1 chromatography 662:675 solid phase chromatography 650:675 solid phase chromatography 650:675 Structural elucidation of the main product (purified by solid phase chromatography) was done by HSQC (2D NMR) which was confirmed as 1-hexyl-α-D-glucopyranoside. 25773197 0 24 theme sucrose 21:27 arg1 Biotransformation 0:16 Biotransformation 0:16 Biotransformation of sucrose into hexyl-α-D-glucopyranoside and -polyglucosides by whole cells of Microbacterium paraoxydans. 0:124 Biotransformation of sucrose into hexyl-α-D-glucopyranoside and -polyglucosides by whole cells of Microbacterium paraoxydans. 25773197 4 25 theme Structural 594:603 arg1 elucidation 605:615 Structural elucidation 594:615 Structural elucidation of the main product (purified by solid phase chromatography) 594:676 Structural elucidation of the main product (purified by solid phase chromatography) was done by HSQC (2D NMR) which was confirmed as 1-hexyl-α-D-glucopyranoside. 25773197 1 26 dep OBJECTIVE 126:134 arg1 determine 139:147 determine 139:147 To determine the transglycosylation activity of cell-bound enzymes from Microbacterium 136:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 5 27 theme total 863:867 arg1 yield 869:873 a total yield 861:873 a total yield of ~44 % 861:882 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 2 28 theme equivalent 468:477 arg1 activity 501:508 equivalent to 8 IU α-glucosidase activity 468:508 equivalent to 8 IU α-glucosidase activity 468:508 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 28 theme equivalent 468:477 arg1 cells 461:465 lyophilized cells 449:465 lyophilized cells (equivalent to 8 IU α-glucosidase activity) 449:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 5 29 theme fed-batch 789:797 arg1 reactor 799:805 a fed-batch reactor 787:805 a fed-batch reactor 787:805 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 2 30 from activity 405:412 arg1 h 420:420 12 h 417:420 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity) 417:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 1 31 theme hexyl-α-D-glucoside 264:282 arg1 synthesis 251:259 the synthesis 247:259 the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor 247:341 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 0 32 theme whole 83:87 arg1 cells 89:93 whole cells 83:93 whole cells of Microbacterium paraoxydans 83:123 Biotransformation of sucrose into hexyl-α-D-glucopyranoside and -polyglucosides by whole cells of Microbacterium paraoxydans. 25773197 2 33 dep RESULTS 344:350 arg1 % 375:375 14.8 % 370:375 14.8 % 370:375 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 33 dep RESULTS 344:350 arg1 yield 363:367 Maximum HG yield 352:367 RESULTS Maximum HG yield (14.8 %) 344:376 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 1 34 theme transglycosylation 153:170 arg1 activity 172:179 the transglycosylation activity 149:179 the transglycosylation activity of cell-bound enzymes from Microbacterium 149:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 6 35 theme tri-glucosides 1011:1024 arg1 Synthesis 980:988 CONCLUSION Synthesis 969:988 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides 969:1024 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 3 36 theme synthesized 516:526 arg1 and-polyglucosides 545:562 The synthesized alkyl-glucosides and-polyglucosides 512:562 The synthesized alkyl-glucosides and-polyglucosides 512:562 The synthesized alkyl-glucosides and-polyglucosides were characterized by ESI-MS. 25773197 4 37 theme product 629:635 arg1 elucidation 605:615 Structural elucidation 594:615 Structural elucidation of the main product (purified by solid phase chromatography) 594:676 Structural elucidation of the main product (purified by solid phase chromatography) was done by HSQC (2D NMR) which was confirmed as 1-hexyl-α-D-glucopyranoside. 25773197 5 38 theme whole 835:839 arg1 cells 841:845 whole cells 835:845 whole cells 835:845 The synthesis was scaled up in a fed-batch reactor, with continuous feeding of whole cells every 6 h and a total yield of ~44 % was obtained for hexyl-glucoside and -polyglucosides under the optimized conditions. 25773197 2 39 theme 0.96 394:397 arg1 activity 405:412 0.96 water activity 394:412 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity) 394:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 6 40 theme di- 1003:1005 arg1 Synthesis 980:988 CONCLUSION Synthesis 969:988 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides 969:1024 CONCLUSION Synthesis of HG, hexyl di- and tri-glucosides has been achieved using a novel method. 25773197 2 41 theme α-glucosidase 487:499 arg1 activity 501:508 equivalent to 8 IU α-glucosidase activity 468:508 equivalent to 8 IU α-glucosidase activity 468:508 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 41 theme α-glucosidase 487:499 arg1 cells 461:465 lyophilized cells 449:465 lyophilized cells (equivalent to 8 IU α-glucosidase activity) 449:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 2 42 from M 442:442 arg1 sucrose 427:433 sucrose 427:433 sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity) 427:509 RESULTS Maximum HG yield (14.8 %) was achieved at 0.96 water activity in 12 h with sucrose at 0.5 M with lyophilized cells (equivalent to 8 IU α-glucosidase activity). 25773197 1 43 from Microbacterium 208:221 arg1 activity 172:179 the transglycosylation activity 149:179 the transglycosylation activity of cell-bound enzymes from Microbacterium 149:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 1 43 from Microbacterium 208:221 arg1 enzymes 195:201 cell-bound enzymes 184:201 cell-bound enzymes from Microbacterium 184:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 25773197 1 44 theme cell-bound 184:193 arg1 enzymes 195:201 cell-bound enzymes 184:201 cell-bound enzymes from Microbacterium 184:221 OBJECTIVE To determine the transglycosylation activity of cell-bound enzymes from Microbacterium paraoxydans to catalyze the synthesis of hexyl-α-D-glucoside (HG) and -polyglucosides using sucrose as a glycosyl donor. 28654715 5 0 dep motifs 956:961 arg1 we 1027:1028 we 1027:1028 we 1027:1028 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 0 dep motifs 956:961 arg1 motifs 956:961 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure 942:1025 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch 942:1047 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 0 dep motifs 956:961 arg1 structure 1017:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 0 dep motifs 956:961 arg1 X-β1,4-[Fucα1,3 963:977 X-β1,4-[Fucα1,3]-Y 963:980 X-β1,4-[Fucα1,3]-Y 963:980 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 4 1 from classes 709:715 arg1 present 689:695 present 689:695 present 689:695 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 0 2 theme Glycoepitopes 62:74 arg1 Range 41:45 a Wide Range 34:45 a Wide Range of Fucosylated Glycoepitopes 34:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 4 3 located found 738:742 arg1 everything 747:756 everything 747:756 everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 747:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 3 located found 738:742 arg2 oligosaccharides 720:735 oligosaccharides 720:735 oligosaccharides 720:735 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 1 4 from development 148:158 arg1 understanding 92:104 The increasing understanding 77:104 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases 77:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 4 5 attach present 689:695 arg1 classes 709:715 numerous classes 700:715 numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 700:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 5 attach present 689:695 arg2 motifs 678:683 These motifs 672:683 These motifs 672:683 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 6 theme blood 800:804 arg1 antigens 812:819 Lewis blood group antigens 794:819 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 5 7 theme F-branch 1040:1047 arg1 motifs 956:961 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure 942:1025 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch 942:1047 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 7 theme F-branch 1040:1047 arg1 structure 1017:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 7 theme F-branch 1040:1047 arg1 X-β1,4-[Fucα1,3 963:977 X-β1,4-[Fucα1,3]-Y 963:980 X-β1,4-[Fucα1,3]-Y 963:980 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 6 8 theme underlying 1197:1206 arg1 architecture 1208:1219 the same underlying architecture 1188:1219 the same underlying architecture 1188:1219 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 6 9 theme wide 1054:1057 arg1 spectrum 1059:1066 The wide spectrum 1050:1066 The wide spectrum of possible modifications of this scaffold 1050:1109 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 1 10 theme basic 233:237 arg1 principles 239:248 the basic principles 229:248 the basic principles governing carbohydrate-protein interactions 229:292 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 4 11 from invertebrates 872:884 arg1 antigens 812:819 Lewis blood group antigens 794:819 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 11 from invertebrates 872:884 arg1 motifs 838:843 unusual motifs 830:843 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 6 12 theme same 1192:1195 arg1 architecture 1208:1219 the same underlying architecture 1188:1219 the same underlying architecture 1188:1219 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 4 13 from present 689:695 arg1 classes 709:715 numerous classes 700:715 numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 700:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 6 14 theme large 1127:1131 arg1 variety 1133:1139 a large variety 1125:1139 a large variety 1125:1139 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 6 14 theme large 1127:1131 arg1 glycoepitopes 1144:1156 glycoepitopes 1144:1156 glycoepitopes 1144:1156 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 6 15 theme glycoepitopes 1144:1156 arg1 variety 1133:1139 a large variety 1125:1139 a large variety 1125:1139 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 6 15 theme glycoepitopes 1144:1156 arg1 glycoepitopes 1144:1156 glycoepitopes 1144:1156 glycoepitopes 1144:1156 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 4 16 theme Lewis 794:798 arg1 antigens 812:819 Lewis blood group antigens 794:819 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 3 17 theme NMR 431:433 arg1 determination 445:457 NMR structure determination 431:457 NMR structure determination based on extensive experimental data 431:494 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 2 18 theme rigidity 391:398 arg1 issue 319:323 A still heavily debated issue 295:323 A still heavily debated issue regarding the recognition process 295:357 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 2 18 theme rigidity 391:398 arg1 degree 366:371 the degree 362:371 the degree of flexibility or rigidity of oligosaccharides 362:418 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 1 19 from role 123:126 arg1 development 148:158 development 148:158 development 148:158 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 5 20 theme -Y-a 1002:1005 arg1 motifs 956:961 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure 942:1025 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch 942:1047 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 20 theme -Y-a 1002:1005 arg1 structure 1017:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 6 21 theme scaffold 1102:1109 arg1 modifications 1080:1092 possible modifications 1071:1092 possible modifications of this scaffold 1071:1109 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 0 22 theme Secondary 2:10 arg1 Element 23:29 A Secondary Structural Element 0:29 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes 0:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 4 23 from amphibians 850:859 arg1 antigens 812:819 Lewis blood group antigens 794:819 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 23 from amphibians 850:859 arg1 motifs 838:843 unusual motifs 830:843 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 24 from bacteria 763:770 arg1 everything 747:756 everything 747:756 everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 747:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 2 25 theme debated 311:317 arg1 issue 319:323 A still heavily debated issue 295:323 A still heavily debated issue regarding the recognition process 295:357 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 2 25 theme debated 311:317 arg1 degree 366:371 the degree 362:371 the degree of flexibility or rigidity of oligosaccharides 362:418 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 2 26 theme flexibility 376:386 arg1 issue 319:323 A still heavily debated issue 295:323 A still heavily debated issue regarding the recognition process 295:357 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 2 26 theme flexibility 376:386 arg1 degree 366:371 the degree 362:371 the degree of flexibility or rigidity of oligosaccharides 362:418 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 3 27 with motifs 570:575 arg1 conformation 592:603 a similar conformation 582:603 a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond 582:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 4 28 theme marine 865:870 arg1 invertebrates 872:884 marine invertebrates 865:884 marine invertebrates 865:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 1 29 theme increasing 81:90 arg1 understanding 92:104 The increasing understanding 77:104 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases 77:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 30 theme wide 170:173 arg1 range 175:179 a wide range 168:179 a wide range of diseases 168:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 31 theme carbohydrate-protein 260:279 arg1 interactions 281:292 carbohydrate-protein interactions 260:292 carbohydrate-protein interactions 260:292 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 3 32 theme trisaccharide 556:568 arg1 motifs 570:575 trisaccharide motifs 556:575 trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond 556:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 4 33 theme unusual 830:836 arg1 motifs 838:843 unusual motifs 830:843 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 3 34 theme experimental 478:489 arg1 data 491:494 extensive experimental data 468:494 extensive experimental data 468:494 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 0 35 theme Structural 12:21 arg1 Element 23:29 A Secondary Structural Element 0:29 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes 0:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 3 36 theme motifs 570:575 arg1 set 549:551 a set 547:551 a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond 547:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 3 36 theme motifs 570:575 arg1 motifs 570:575 trisaccharide motifs 556:575 trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond 556:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 4 37 theme numerous 700:707 arg1 classes 709:715 numerous classes 700:715 numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 700:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 3 38 theme database 509:516 arg1 searches 518:525 database searches 509:525 database searches 509:525 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 2 39 theme oligosaccharides 403:418 arg1 rigidity 391:398 rigidity 391:398 rigidity 391:398 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 2 39 theme oligosaccharides 403:418 arg1 flexibility 376:386 flexibility 376:386 flexibility 376:386 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 4 40 located present 689:695 arg1 classes 709:715 numerous classes 700:715 numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 700:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 4 40 located present 689:695 arg2 motifs 678:683 These motifs 672:683 These motifs 672:683 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 3 41 theme structure 435:443 arg1 determination 445:457 NMR structure determination 431:457 NMR structure determination based on extensive experimental data 431:494 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 1 42 theme diseases 184:191 arg1 range 175:179 a wide range 168:179 a wide range of diseases 168:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 6 43 theme modifications 1080:1092 arg1 spectrum 1059:1066 The wide spectrum 1050:1066 The wide spectrum of possible modifications of this scaffold 1050:1109 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 3 44 theme C-H⋅⋅⋅O 649:655 arg1 bond 666:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 3 45 theme extensive 468:476 arg1 data 491:494 extensive experimental data 468:494 extensive experimental data 468:494 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 6 46 theme possible 1071:1078 arg1 modifications 1080:1092 possible modifications 1071:1092 possible modifications of this scaffold 1071:1109 The wide spectrum of possible modifications of this scaffold points toward a large variety of glycoepitopes, which nature generated using the same underlying architecture. 28654715 5 47 theme trisaccharide 898:910 arg1 motifs 912:917 trisaccharide motifs 898:917 trisaccharide motifs 898:917 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 3 48 theme hydrogen 657:664 arg1 bond 666:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 3 49 theme similar 584:590 arg1 conformation 592:603 a similar conformation 582:603 a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond 582:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 2 50 theme recognition 339:349 arg1 process 351:357 the recognition process 335:357 the recognition process 335:357 A still heavily debated issue regarding the recognition process is the degree of flexibility or rigidity of oligosaccharides. 28654715 0 51 theme Wide 36:39 arg1 Range 41:45 a Wide Range 34:45 a Wide Range of Fucosylated Glycoepitopes 34:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 1 52 from interest 217:224 arg1 principles 239:248 the basic principles 229:248 the basic principles governing carbohydrate-protein interactions 229:292 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 53 theme essential 113:121 arg1 role 123:126 the essential role 109:126 the essential role of carbohydrates in development 109:158 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 54 from understanding 92:104 arg1 development 148:158 development 148:158 development 148:158 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 54 from understanding 92:104 arg1 range 175:179 a wide range 168:179 a wide range of diseases 168:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 5 55 theme secondary 1007:1015 arg1 motifs 956:961 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure 942:1025 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch 942:1047 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 55 theme secondary 1007:1015 arg1 structure 1017:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 4 56 theme group 806:810 arg1 antigens 812:819 Lewis blood group antigens 794:819 Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 794:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 28654715 1 57 theme role 123:126 arg1 understanding 92:104 The increasing understanding 77:104 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases 77:191 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 3 58 theme non-conventional 632:647 arg1 bond 666:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 a non-conventional C-H⋅⋅⋅O hydrogen bond 630:669 Combining NMR structure determination based on extensive experimental data with DFT and database searches, we have identified a set of trisaccharide motifs with a similar conformation that is characterized by a non-conventional C-H⋅⋅⋅O hydrogen bond. 28654715 5 59 theme consensus 946:954 arg1 motifs 956:961 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure 942:1025 the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch 942:1047 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 59 theme consensus 946:954 arg1 structure 1017:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 X-β1,3-[Fucα1,4]-Y-a secondary structure 986:1025 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 59 theme consensus 946:954 arg1 X-β1,4-[Fucα1,3 963:977 X-β1,4-[Fucα1,3]-Y 963:980 X-β1,4-[Fucα1,3]-Y 963:980 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 0 60 theme Fucosylated 50:60 arg1 Glycoepitopes 62:74 Fucosylated Glycoepitopes 50:74 Fucosylated Glycoepitopes 50:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 1 61 theme carbohydrates 131:143 arg1 role 123:126 the essential role 109:126 the essential role of carbohydrates in development 109:158 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 1 62 theme growing 209:215 arg1 interest 217:224 a rapidly growing interest 199:224 a rapidly growing interest in the basic principles governing carbohydrate-protein interactions 199:292 The increasing understanding of the essential role of carbohydrates in development, and in a wide range of diseases fuels a rapidly growing interest in the basic principles governing carbohydrate-protein interactions. 28654715 0 63 from Element 23:29 arg1 Range 41:45 a Wide Range 34:45 a Wide Range of Fucosylated Glycoepitopes 34:74 A Secondary Structural Element in a Wide Range of Fucosylated Glycoepitopes. 28654715 5 64 theme motifs 912:917 arg1 set 891:893 The set 887:893 The set of trisaccharide motifs 887:917 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 5 64 theme motifs 912:917 arg1 motifs 912:917 trisaccharide motifs 898:917 trisaccharide motifs 898:917 The set of trisaccharide motifs can be summarized with the consensus motifs X-β1,4-[Fucα1,3]-Y and X-β1,3-[Fucα1,4]-Y-a secondary structure we name [3,4]F-branch. 28654715 4 65 theme oligosaccharides 720:735 arg1 classes 709:715 numerous classes 700:715 numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates 700:884 These motifs are present in numerous classes of oligosaccharides, found in everything from bacteria to mammals, including Lewis blood group antigens but also unusual motifs from amphibians and marine invertebrates. 25231986 8 0 theme core 1500:1503 arg1 substitution 1473:1484 exactly one arabinosyl substitution 1450:1484 exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1450:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 1 theme single 1668:1673 arg1 primer 1687:1692 a single arabinosyl "primer 1666:1692 a single arabinosyl "primer 1666:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 4 2 from backbone 654:661 arg1 LAM 666:668 LAM 666:668 LAM 666:668 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 9 3 from LAM 1823:1825 arg1 present 1805:1811 present 1805:1811 present 1805:1811 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 3 from LAM 1823:1825 arg1 chain 1752:1756 only a single arabinan chain 1729:1756 only a single arabinan chain attached near the middle of the mannan core 1729:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 4 theme updated 1844:1850 arg1 model 1860:1864 an updated working model 1841:1864 an updated working model of the biosynthetic pathway of LAM and LM 1841:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 6 5 theme α- 871:872 arg1 1,6 874:876 1,6 874:876 1,6 874:876 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 6 5 theme α- 871:872 arg1 -mannosyltransferase 878:897 the α-(1,6)-mannosyltransferase 867:897 the α-(1,6)-mannosyltransferase used late in LM elongation 867:924 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 0 6 attach attached 27:34 arg1 core 73:76 the phosphatidylinositol mannosyl core 39:76 the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan 39:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 6 attach attached 27:34 arg2 chain 18:22 A single arabinan chain 0:22 A single arabinan chain 0:22 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 6 7 attach attached 988:995 arg2 mannan 973:978 the mannan 969:978 the mannan that is attached to inositol 969:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 6 7 attach attached 988:995 arg1 inositol 1000:1007 inositol 1000:1007 inositol 1000:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 6 8 theme single 1125:1130 arg1 residues 1148:1155 single α-mannopyranose residues 1125:1155 single α-mannopyranose residues at O-2 1125:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 0 9 theme cell 122:125 arg1 glycoconjugate 136:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 9 theme cell 122:125 arg1 lipoarabinomannan 152:168 lipoarabinomannan 152:168 lipoarabinomannan 152:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 9 10 located present 1805:1811 arg2 chain 1752:1756 only a single arabinan chain 1729:1756 only a single arabinan chain attached near the middle of the mannan core 1729:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 10 located present 1805:1811 arg2 present 1805:1811 present 1805:1811 present 1805:1811 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 10 located present 1805:1811 arg1 LAM 1823:1825 mature LAM 1816:1825 mature LAM 1816:1825 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 6 11 theme unbranched 1017:1026 arg1 residues 1048:1055 5-7 unbranched α-6-linked-mannosyl residues 1013:1055 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1013:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 4 12 theme chains 633:638 arg1 number 614:619 the number 610:619 the number of arabinan chains 610:638 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 4 12 theme chains 633:638 arg1 structure 590:598 the branching structure 576:598 the branching structure of LM 576:604 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 0 13 theme glycoconjugate 136:149 arg1 core 73:76 the phosphatidylinositol mannosyl core 39:76 the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan 39:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 9 14 theme LAM 1897:1899 arg1 pathway 1886:1892 the biosynthetic pathway 1869:1892 the biosynthetic pathway of LAM and LM 1869:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 6 15 theme LM 912:913 arg1 elongation 915:924 LM elongation 912:924 LM elongation 912:924 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 0 16 theme immunomodulatory 91:106 arg1 glycoconjugate 136:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 16 theme immunomodulatory 91:106 arg1 lipoarabinomannan 152:168 lipoarabinomannan 152:168 lipoarabinomannan 152:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 9 17 theme LM 1905:1906 arg1 pathway 1886:1892 the biosynthetic pathway 1869:1892 the biosynthetic pathway of LAM and LM 1869:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 7 18 theme mannan 1220:1225 arg1 chain 1227:1231 the α-6-linked mannan chain 1205:1231 the α-6-linked mannan chain 1205:1231 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 8 19 from core 1381:1384 arg1 ΔembC 1414:1418 ΔembC 1414:1418 ΔembC 1414:1418 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 19 from core 1381:1384 arg1 mutants 1405:1411 two other mutants 1395:1411 two other mutants (ΔembC and ΔMSMEG_4247) 1395:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 19 from core 1381:1384 arg1 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 20 theme mannan 973:978 arg1 region 959:964 the reducing end region 942:964 the reducing end region of the mannan that is attached to inositol 942:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 5 21 theme Mycobacterium 758:770 arg1 smegmatis 772:780 Mycobacterium smegmatis 758:780 Mycobacterium smegmatis 758:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 1 22 theme mannose 354:360 arg1 oligosaccharides 334:349 oligosaccharides 334:349 oligosaccharides of mannose 334:360 Lipoarabinomannan (LAM) is composed of a phosphatidylinositol anchor followed by a mannan followed by an arabinan that may be capped with various motifs including oligosaccharides of mannose. 25231986 6 23 theme end 955:957 arg1 region 959:964 the reducing end region 942:964 the reducing end region of the mannan that is attached to inositol 942:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 9 24 theme arabinan 1743:1750 arg1 present 1805:1811 present 1805:1811 present 1805:1811 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 24 theme arabinan 1743:1750 arg1 chain 1752:1756 only a single arabinan chain 1729:1756 only a single arabinan chain attached near the middle of the mannan core 1729:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 8 25 theme number 1338:1343 arg1 Analysis 1322:1329 Analysis 1322:1329 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1322:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 5 26 theme different 717:725 arg1 mutants 747:753 three different glycosyltransferase mutants 711:753 three different glycosyltransferase mutants of Mycobacterium smegmatis 711:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 4 27 theme fundamental 548:558 arg1 questions 560:568 fundamental questions 548:568 fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM 548:668 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 7 28 theme α-6-linked 1209:1218 arg1 chain 1227:1231 the α-6-linked mannan chain 1205:1231 the α-6-linked mannan chain 1205:1231 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 8 29 theme arabinans 1348:1356 arg1 number 1338:1343 the number 1334:1343 the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1334:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 7 30 theme penultimate 1301:1311 arg1 residue 1313:1319 the penultimate residue 1297:1319 the penultimate residue 1297:1319 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 0 31 theme arabinan 9:16 arg1 chain 18:22 A single arabinan chain 0:22 A single arabinan chain 0:22 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 5 32 used used 787:790 arg2 molecules 689:697 LM and LAM molecules 678:697 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis 678:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 8 33 theme arabinosylation 1523:1537 arg1 suggestive 1505:1514 suggestive 1505:1514 suggestive 1505:1514 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 34 link α-6-linked 1082:1091 arg1 residues 1102:1109 two or three α-6-linked mannosyl residues 1069:1109 two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1069:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 8 35 theme mannan 1374:1379 arg1 core 1381:1384 the mannan core 1370:1384 the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1370:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 9 36 theme mannan 1790:1795 arg1 core 1797:1800 the mannan core 1786:1800 the mannan core 1786:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 0 37 theme phosphatidylinositol 43:62 arg1 core 73:76 the phosphatidylinositol mannosyl core 39:76 the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan 39:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 6 38 theme MSMEG_4241 838:847 arg1 mutant 849:854 an MSMEG_4241 mutant 835:854 an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation 835:924 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 8 39 theme LM 1551:1552 arg1 precursor 1554:1562 a linear LM precursor 1542:1562 a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1542:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 40 theme α-6-linked 1082:1091 arg1 residues 1102:1109 two or three α-6-linked mannosyl residues 1069:1109 two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1069:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 8 41 theme other 1399:1403 arg1 ΔembC 1414:1418 ΔembC 1414:1418 ΔembC 1414:1418 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 41 theme other 1399:1403 arg1 mutants 1405:1411 two other mutants 1395:1411 two other mutants (ΔembC and ΔMSMEG_4247) 1395:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 41 theme other 1399:1403 arg1 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 3 42 theme LAM 504:506 arg1 structure 470:478 the structure 466:478 the structure 466:478 Both the structure and the biosynthesis of LAM have been studied extensively. 25231986 3 42 theme LAM 504:506 arg1 biosynthesis 488:499 the biosynthesis 484:499 the biosynthesis of LAM 484:506 Both the structure and the biosynthesis of LAM have been studied extensively. 25231986 8 43 theme core 1638:1641 arg1 mannosylation 1617:1629 additional mannosylation 1606:1629 additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1606:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 44 gly mannosylation 1617:1629 arg1 core 1638:1641 the core 1634:1641 the core 1634:1641 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 44 gly mannosylation 1617:1629 arg1 arabinosylation 1647:1661 arabinosylation 1647:1661 arabinosylation 1647:1661 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 45 theme mannosyl 1576:1583 arg1 residues 1585:1592 ∼10-12 mannosyl residues 1569:1592 ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1569:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 2 46 theme related 365:371 arg1 lipomannan 382:391 lipomannan 382:391 lipomannan (LM) 382:396 A related polymer, lipomannan (LM), is composed of only the phosphatidylinositol and mannan core. 25231986 2 46 theme related 365:371 arg1 polymer 373:379 A related polymer 363:379 A related polymer 363:379 A related polymer, lipomannan (LM), is composed of only the phosphatidylinositol and mannan core. 25231986 8 47 theme arabinosylation 1647:1661 arg1 mannosylation 1617:1629 additional mannosylation 1606:1629 additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1606:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 48 theme additional 1606:1615 arg1 mannosylation 1617:1629 additional mannosylation 1606:1629 additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1606:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 49 theme arabinosyl 1675:1684 arg1 primer 1687:1692 a single arabinosyl "primer 1666:1692 a single arabinosyl "primer 1666:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 9 50 theme core 1797:1800 arg1 middle 1776:1781 the middle 1772:1781 the middle of the mannan core 1772:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 51 theme working 1852:1858 arg1 model 1860:1864 an updated working model 1841:1864 an updated working model of the biosynthetic pathway of LAM and LM 1841:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 8 52 from mutants 1405:1411 arg1 core 1381:1384 the mannan core 1370:1384 the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1370:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 0 53 theme mannosyl 64:71 arg1 core 73:76 the phosphatidylinositol mannosyl core 39:76 the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan 39:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 8 54 theme primer 1687:1692 arg1 core 1638:1641 the core 1634:1641 the core 1634:1641 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 54 theme primer 1687:1692 arg1 arabinosylation 1647:1661 arabinosylation 1647:1661 arabinosylation 1647:1661 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 55 theme reducing 946:953 arg1 region 959:964 the reducing end region 942:964 the reducing end region of the mannan that is attached to inositol 942:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 6 56 theme α-mannopyranose 1132:1146 arg1 residues 1148:1155 single α-mannopyranose residues 1125:1155 single α-mannopyranose residues at O-2 1125:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 4 57 theme mannan 647:652 arg1 backbone 654:661 the mannan backbone 643:661 the mannan backbone in LAM 643:668 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 0 58 theme envelope 127:134 arg1 glycoconjugate 136:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 58 theme envelope 127:134 arg1 lipoarabinomannan 152:168 lipoarabinomannan 152:168 lipoarabinomannan 152:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 7 59 link α-6-linked 1209:1218 arg1 chain 1227:1231 the α-6-linked mannan chain 1205:1231 the α-6-linked mannan chain 1205:1231 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 9 60 from present 1805:1811 arg1 LAM 1823:1825 mature LAM 1816:1825 mature LAM 1816:1825 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 61 theme biosynthetic 1873:1884 arg1 pathway 1886:1892 the biosynthetic pathway 1869:1892 the biosynthetic pathway of LAM and LM 1869:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 5 62 theme LAM 685:687 arg1 molecules 689:697 LM and LAM molecules 678:697 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis 678:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 8 63 from LM 1389:1390 arg1 ΔembC 1414:1418 ΔembC 1414:1418 ΔembC 1414:1418 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 63 from LM 1389:1390 arg1 mutants 1405:1411 two other mutants 1395:1411 two other mutants (ΔembC and ΔMSMEG_4247) 1395:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 63 from LM 1389:1390 arg1 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 1 64 theme phosphatidylinositol 212:231 arg1 anchor 233:238 a phosphatidylinositol anchor 210:238 a phosphatidylinositol anchor followed by a mannan followed by an arabinan that may be capped with various motifs including oligosaccharides of mannose 210:360 Lipoarabinomannan (LAM) is composed of a phosphatidylinositol anchor followed by a mannan followed by an arabinan that may be capped with various motifs including oligosaccharides of mannose. 25231986 4 65 theme arabinan 624:631 arg1 chains 633:638 arabinan chains 624:638 arabinan chains 624:638 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 7 66 theme mannosyl 1186:1193 arg1 residues 1195:1202 these branched mannosyl residues 1171:1202 these branched mannosyl residues 1171:1202 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 5 67 theme LM 678:679 arg1 molecules 689:697 LM and LAM molecules 678:697 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis 678:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 8 68 with precursor 1554:1562 arg1 residues 1585:1592 ∼10-12 mannosyl residues 1569:1592 ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1569:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 5 69 theme glycosyltransferase 727:745 arg1 mutants 747:753 three different glycosyltransferase mutants 711:753 three different glycosyltransferase mutants of Mycobacterium smegmatis 711:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 8 70 theme arabinosyl 1462:1471 arg1 substitution 1473:1484 exactly one arabinosyl substitution 1450:1484 exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1450:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 71 theme ∼10-12 1569:1574 arg1 residues 1585:1592 ∼10-12 mannosyl residues 1569:1592 ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1569:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 0 72 theme major 85:89 arg1 glycoconjugate 136:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 72 theme major 85:89 arg1 lipoarabinomannan 152:168 lipoarabinomannan 152:168 lipoarabinomannan 152:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 9 73 theme single 1736:1741 arg1 present 1805:1811 present 1805:1811 present 1805:1811 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 73 theme single 1736:1741 arg1 chain 1752:1756 only a single arabinan chain 1729:1756 only a single arabinan chain attached near the middle of the mannan core 1729:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 8 74 theme mannan 1493:1498 arg1 core 1500:1503 the mannan core 1489:1503 the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1489:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 75 from O-2 1160:1162 arg1 residues 1148:1155 single α-mannopyranose residues 1125:1155 single α-mannopyranose residues at O-2 1125:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 0 76 theme mycobacterial 108:120 arg1 glycoconjugate 136:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 the major immunomodulatory mycobacterial cell envelope glycoconjugate 81:149 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 0 76 theme mycobacterial 108:120 arg1 lipoarabinomannan 152:168 lipoarabinomannan 152:168 lipoarabinomannan 152:168 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 7 77 theme residue 1313:1319 arg1 O-6 1290:1292 O-6 1290:1292 O-6 of the penultimate residue 1290:1319 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 6 78 contain has 1009:1011 arg2 residues 1048:1055 5-7 unbranched α-6-linked-mannosyl residues 1013:1055 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1013:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 6 78 contain has 1009:1011 arg1 region 959:964 the reducing end region 942:964 the reducing end region of the mannan that is attached to inositol 942:1007 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 8 79 attach attached 1358:1365 arg1 core 1381:1384 the mannan core 1370:1384 the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1370:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 79 attach attached 1358:1365 arg2 arabinans 1348:1356 arabinans 1348:1356 arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1348:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 0 80 theme single 2:7 arg1 chain 18:22 A single arabinan chain 0:22 A single arabinan chain 0:22 A single arabinan chain is attached to the phosphatidylinositol mannosyl core of the major immunomodulatory mycobacterial cell envelope glycoconjugate, lipoarabinomannan. 25231986 6 81 theme α-6-linked-mannosyl 1028:1046 arg1 residues 1048:1055 5-7 unbranched α-6-linked-mannosyl residues 1013:1055 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1013:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 2 82 theme phosphatidylinositol 423:442 arg1 core 455:458 only the phosphatidylinositol and mannan core 414:458 only the phosphatidylinositol and mannan core 414:458 A related polymer, lipomannan (LM), is composed of only the phosphatidylinositol and mannan core. 25231986 8 83 dep demonstrated 1437:1448 arg1 " 1694:1694 " 1694:1694 " 1694:1694 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 9 84 attach present 1805:1811 arg2 chain 1752:1756 only a single arabinan chain 1729:1756 only a single arabinan chain attached near the middle of the mannan core 1729:1800 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 84 attach present 1805:1811 arg2 present 1805:1811 present 1805:1811 present 1805:1811 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 84 attach present 1805:1811 arg1 LAM 1823:1825 mature LAM 1816:1825 mature LAM 1816:1825 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 9 85 theme pathway 1886:1892 arg1 model 1860:1864 an updated working model 1841:1864 an updated working model of the biosynthetic pathway of LAM and LM 1841:1906 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 5 86 theme smegmatis 772:780 arg1 mutants 747:753 three different glycosyltransferase mutants 711:753 three different glycosyltransferase mutants of Mycobacterium smegmatis 711:780 LM and LAM molecules produced by three different glycosyltransferase mutants of Mycobacterium smegmatis were used here to investigate these questions. 25231986 8 87 theme linear 1544:1549 arg1 precursor 1554:1562 a linear LM precursor 1542:1562 a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1542:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 7 88 theme branched 1177:1184 arg1 residues 1195:1202 these branched mannosyl residues 1171:1202 these branched mannosyl residues 1171:1202 After these branched mannosyl residues, the α-6-linked mannan chain is terminated with an α-mannopyranose at O-2 rather than O-6 of the penultimate residue. 25231986 1 89 theme various 309:315 arg1 oligosaccharides 334:349 oligosaccharides 334:349 oligosaccharides of mannose 334:360 Lipoarabinomannan (LAM) is composed of a phosphatidylinositol anchor followed by a mannan followed by an arabinan that may be capped with various motifs including oligosaccharides of mannose. 25231986 1 89 theme various 309:315 arg1 motifs 317:322 various motifs 309:322 various motifs including oligosaccharides of mannose 309:360 Lipoarabinomannan (LAM) is composed of a phosphatidylinositol anchor followed by a mannan followed by an arabinan that may be capped with various motifs including oligosaccharides of mannose. 25231986 8 90 theme suggestive 1505:1514 arg1 core 1500:1503 the mannan core 1489:1503 the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1489:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 2 91 theme mannan 448:453 arg1 core 455:458 only the phosphatidylinositol and mannan core 414:458 only the phosphatidylinositol and mannan core 414:458 A related polymer, lipomannan (LM), is composed of only the phosphatidylinositol and mannan core. 25231986 8 92 theme precursor 1554:1562 arg1 arabinosylation 1523:1537 the arabinosylation 1519:1537 the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer 1519:1692 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 93 dep mutants 1405:1411 arg1 ΔembC 1414:1418 ΔembC 1414:1418 ΔembC 1414:1418 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 93 dep mutants 1405:1411 arg1 mutants 1405:1411 two other mutants 1395:1411 two other mutants (ΔembC and ΔMSMEG_4247) 1395:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 8 93 dep mutants 1405:1411 arg1 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 ΔMSMEG_4247 1424:1434 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 6 94 theme mannosyl 1093:1100 arg1 residues 1102:1109 two or three α-6-linked mannosyl residues 1069:1109 two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2 1069:1162 Using an MSMEG_4241 mutant that lacks the α-(1,6)-mannosyltransferase used late in LM elongation, we showed that the reducing end region of the mannan that is attached to inositol has 5-7 unbranched α-6-linked-mannosyl residues followed by two or three α-6-linked mannosyl residues branched with single α-mannopyranose residues at O-2. 25231986 4 95 theme LM 603:604 arg1 number 614:619 the number 610:619 the number of arabinan chains 610:638 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 4 95 theme LM 603:604 arg1 structure 590:598 the branching structure 576:598 the branching structure of LM 576:604 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 8 96 theme LM 1389:1390 arg1 core 1381:1384 the mannan core 1370:1384 the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) 1370:1435 Analysis of the number of arabinans attached to the mannan core of LM in two other mutants (ΔembC and ΔMSMEG_4247) demonstrated exactly one arabinosyl substitution of the mannan core suggestive of the arabinosylation of a linear LM precursor with ∼10-12 mannosyl residues followed by additional mannosylation of the core and arabinosylation of a single arabinosyl "primer." 25231986 9 97 theme mature 1816:1821 arg1 LAM 1823:1825 mature LAM 1816:1825 mature LAM 1816:1825 Thus, these studies suggest that only a single arabinan chain attached near the middle of the mannan core is present in mature LAM and allow for an updated working model of the biosynthetic pathway of LAM and LM. 25231986 4 98 theme branching 580:588 arg1 structure 590:598 the branching structure 576:598 the branching structure of LM 576:604 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 25231986 4 99 from questions 560:568 arg1 backbone 654:661 the mannan backbone 643:661 the mannan backbone in LAM 643:668 However, fundamental questions about the branching structure of LM and the number of arabinan chains on the mannan backbone in LAM remain. 29124933 4 0 theme l-fucosyl-d-xylose 664:681 arg1 unit 683:686 the l-fucosyl-d-xylose unit and construction 660:703 unit 683:686 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 1 1 theme natural 128:134 arg1 Polycavernosides 78:93 Polycavernosides A and B 78:101 Polycavernosides A and B 78:101 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 1 theme natural 128:134 arg1 products 136:143 glycosidic macrolide natural products 107:143 glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai) 107:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 2 theme alga 216:219 arg1 edulis 232:237 the edible red alga Gracilaria edulis 201:237 the edible red alga Gracilaria edulis (Polycavernosa tsudai) 201:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 2 theme alga 216:219 arg1 tsudai 254:259 tsudai 254:259 tsudai 254:259 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 3 3 theme fragments 469:477 arg1 syntheses 444:452 the catalytic asymmetric syntheses 419:452 the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization 419:629 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 1 4 theme Gracilaria 221:230 arg1 edulis 232:237 the edible red alga Gracilaria edulis 201:237 the edible red alga Gracilaria edulis (Polycavernosa tsudai) 201:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 4 theme Gracilaria 221:230 arg1 tsudai 254:259 tsudai 254:259 tsudai 254:259 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 3 5 theme Kiyooka 517:523 arg1 steps 551:555 the key steps 543:555 the key steps 543:555 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 5 theme Kiyooka 517:523 arg1 reaction 504:511 hetero-Diels-Alder reaction 485:511 hetero-Diels-Alder reaction 485:511 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 5 theme Kiyooka 517:523 arg1 reaction 531:538 Kiyooka aldol reaction 517:538 Kiyooka aldol reaction 517:538 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 0 6 theme Red 68:70 arg1 Alga 72:75 Red Alga 68:75 Red Alga 68:75 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 3 7 theme aldol 525:529 arg1 steps 551:555 the key steps 543:555 the key steps 543:555 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 7 theme aldol 525:529 arg1 reaction 504:511 hetero-Diels-Alder reaction 485:511 hetero-Diels-Alder reaction 485:511 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 7 theme aldol 525:529 arg1 reaction 531:538 Kiyooka aldol reaction 517:538 Kiyooka aldol reaction 517:538 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 8 theme asymmetric 433:442 arg1 syntheses 444:452 the catalytic asymmetric syntheses 419:452 the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization 419:629 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 4 9 theme total 769:773 arg1 synthesis 775:783 the total synthesis 765:783 the total synthesis of polycavernosides A and B 765:811 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 10 dep polycavernosides 788:803 arg1 polycavernosides 788:803 polycavernosides A and B 788:811 polycavernosides A and B 788:811 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 10 dep polycavernosides 788:803 arg1 B 811:811 B 811:811 B 811:811 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 10 dep polycavernosides 788:803 arg1 A 805:805 A 805:805 A 805:805 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 2 11 theme polycavernosides 282:297 arg1 synthesis 269:277 Total synthesis 263:277 Total synthesis of polycavernosides A and B 263:305 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 3 12 theme core 386:389 arg1 structure 391:399 the macrolactone core structure 369:399 the macrolactone core structure 369:399 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 0 13 theme Total 0:4 arg1 Toxins 56:61 Two Lethal Toxins 45:61 Two Lethal Toxins from Red Alga 45:75 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 13 theme Total 0:4 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Polycavernosides A and B 0:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 3 14 theme key 547:549 arg1 steps 551:555 the key steps 543:555 the key steps 543:555 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 14 theme key 547:549 arg1 reaction 504:511 hetero-Diels-Alder reaction 485:511 hetero-Diels-Alder reaction 485:511 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 14 theme key 547:549 arg1 reaction 531:538 Kiyooka aldol reaction 517:538 Kiyooka aldol reaction 517:538 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 2 15 theme convergent 331:340 arg1 approach 342:349 a convergent approach 329:349 a convergent approach 329:349 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 0 16 theme Polycavernosides 19:34 arg1 Toxins 56:61 Two Lethal Toxins 45:61 Two Lethal Toxins from Red Alga 45:75 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 16 theme Polycavernosides 19:34 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Polycavernosides A and B 0:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 4 17 theme Stille 739:744 arg1 coupling 746:753 Stille coupling 739:753 Stille coupling 739:753 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 3 18 theme catalytic 423:431 arg1 syntheses 444:452 the catalytic asymmetric syntheses 419:452 the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization 419:629 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 4 19 theme side 720:723 arg1 chain 725:729 the polyene side chain 708:729 the polyene side chain 708:729 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 2 20 theme Total 263:267 arg1 synthesis 269:277 Total synthesis 263:277 Total synthesis of polycavernosides A and B 263:305 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 1 21 theme fatal 176:180 arg1 poisoning 188:196 fatal human poisoning 176:196 fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai) 176:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 3 22 theme key 465:467 arg1 fragments 469:477 the two key fragments 457:477 the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization 457:629 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 4 23 with glycosylation 641:653 arg1 unit 683:686 the l-fucosyl-d-xylose unit and construction 660:703 unit 683:686 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 23 with glycosylation 641:653 arg1 construction 692:703 the l-fucosyl-d-xylose unit and construction 660:703 construction 692:703 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 3 24 theme structure 391:399 arg1 synthesis 356:364 The synthesis 352:364 The synthesis of the macrolactone core structure 352:399 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 1 25 theme human 182:186 arg1 poisoning 188:196 fatal human poisoning 176:196 fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai) 176:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 3 26 theme macrolactone 373:384 arg1 structure 391:399 the macrolactone core structure 369:399 the macrolactone core structure 369:399 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 27 theme Keck 607:610 arg1 macrolactonization 612:629 Keck macrolactonization 607:629 Keck macrolactonization 607:629 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 1 28 theme macrolide 118:126 arg1 Polycavernosides 78:93 Polycavernosides A and B 78:101 Polycavernosides A and B 78:101 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 28 theme macrolide 118:126 arg1 products 136:143 glycosidic macrolide natural products 107:143 glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai) 107:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 4 29 theme polycavernosides 788:803 arg1 synthesis 775:783 the total synthesis 765:783 the total synthesis of polycavernosides A and B 765:811 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 30 theme chain 725:729 arg1 unit 683:686 the l-fucosyl-d-xylose unit and construction 660:703 unit 683:686 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 4 30 theme chain 725:729 arg1 construction 692:703 the l-fucosyl-d-xylose unit and construction 660:703 construction 692:703 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 0 31 dep Polycavernosides 19:34 arg1 Polycavernosides 19:34 Polycavernosides A and B 19:42 Polycavernosides A and B 19:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 31 dep Polycavernosides 19:34 arg1 B 42:42 B 42:42 B 42:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 31 dep Polycavernosides 19:34 arg1 A 36:36 A 36:36 A 36:36 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 2 32 dep polycavernosides 282:297 arg1 B 305:305 B 305:305 B 305:305 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 2 32 dep polycavernosides 282:297 arg1 A 299:299 A 299:299 A 299:299 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 2 32 dep polycavernosides 282:297 arg1 polycavernosides 282:297 polycavernosides A and B 282:305 polycavernosides A and B 282:305 Total synthesis of polycavernosides A and B has been achieved via a convergent approach. 29124933 0 33 from Alga 72:75 arg1 Toxins 56:61 Two Lethal Toxins 45:61 Two Lethal Toxins from Red Alga 45:75 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 33 from Alga 72:75 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Polycavernosides A and B 0:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 1 34 dep Polycavernosides 78:93 arg1 B 101:101 B 101:101 B 101:101 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 34 dep Polycavernosides 78:93 arg1 A 95:95 A 95:95 A 95:95 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 34 dep Polycavernosides 78:93 arg1 Polycavernosides 78:93 Polycavernosides A and B 78:101 Polycavernosides A and B 78:101 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 4 35 theme polyene 712:718 arg1 chain 725:729 the polyene side chain 708:729 the polyene side chain 708:729 Finally, glycosylation with the l-fucosyl-d-xylose unit and construction of the polyene side chain through Stille coupling completed the total synthesis of polycavernosides A and B. 29124933 0 36 theme Lethal 49:54 arg1 Toxins 56:61 Two Lethal Toxins 45:61 Two Lethal Toxins from Red Alga 45:75 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 0 36 theme Lethal 49:54 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Polycavernosides A and B 0:42 Total Synthesis of Polycavernosides A and B, Two Lethal Toxins from Red Alga. 29124933 1 37 theme glycosidic 107:116 arg1 Polycavernosides 78:93 Polycavernosides A and B 78:101 Polycavernosides A and B 78:101 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 37 theme glycosidic 107:116 arg1 products 136:143 glycosidic macrolide natural products 107:143 glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai) 107:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 38 theme edible 205:210 arg1 edulis 232:237 the edible red alga Gracilaria edulis 201:237 the edible red alga Gracilaria edulis (Polycavernosa tsudai) 201:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 38 theme edible 205:210 arg1 tsudai 254:259 tsudai 254:259 tsudai 254:259 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 3 39 theme Suzuki-Miyaura 578:591 arg1 coupling 593:600 Suzuki-Miyaura coupling 578:600 Suzuki-Miyaura coupling 578:600 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 40 theme hetero-Diels-Alder 485:502 arg1 reaction 504:511 hetero-Diels-Alder reaction 485:511 hetero-Diels-Alder reaction 485:511 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 40 theme hetero-Diels-Alder 485:502 arg1 steps 551:555 the key steps 543:555 the key steps 543:555 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 3 40 theme hetero-Diels-Alder 485:502 arg1 reaction 531:538 Kiyooka aldol reaction 517:538 Kiyooka aldol reaction 517:538 The synthesis of the macrolactone core structure is highlighted by the catalytic asymmetric syntheses of the two key fragments using hetero-Diels-Alder reaction and Kiyooka aldol reaction as the key steps, their union through Suzuki-Miyaura coupling, and Keck macrolactonization. 29124933 1 41 theme red 212:214 arg1 edulis 232:237 the edible red alga Gracilaria edulis 201:237 the edible red alga Gracilaria edulis (Polycavernosa tsudai) 201:260 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 29124933 1 41 theme red 212:214 arg1 tsudai 254:259 tsudai 254:259 tsudai 254:259 Polycavernosides A and B are glycosidic macrolide natural products isolated as the toxins causing fatal human poisoning by the edible red alga Gracilaria edulis (Polycavernosa tsudai). 25230686 6 0 theme complex 1064:1070 arg1 formation 1072:1080 homomeric and heteromeric protein complex formation 1030:1080 homomeric and heteromeric protein complex formation 1030:1080 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 7 1 theme glycosylation 1238:1250 arg1 defect 1252:1257 the GnTI-dependent glycosylation defect 1219:1257 the GnTI-dependent glycosylation defect 1219:1257 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 8 2 from cis/medial-Golgi 1397:1412 arg1 plants 1417:1422 plants 1417:1422 plants 1417:1422 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 4 3 theme Nicotiana 776:784 arg1 benthamiana 786:796 Nicotiana benthamiana 776:796 Nicotiana benthamiana 776:796 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 6 4 theme GnTI 996:999 arg1 region 986:991 the stem region 977:991 the stem region of GnTI 977:999 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 7 5 theme sub-Golgi 1169:1177 arg1 localization 1179:1190 altered sub-Golgi localization 1161:1190 altered sub-Golgi localization 1161:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 4 6 theme protein 639:645 arg1 domains 647:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 0 7 theme Golgi 94:98 arg1 subcompartmentation 100:118 its Golgi subcompartmentation 90:118 its Golgi subcompartmentation 90:118 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 7 8 from thaliana 1122:1129 arg1 variant 1148:1154 a chimeric GnTI variant 1132:1154 a chimeric GnTI variant with altered sub-Golgi localization 1132:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 7 8 from thaliana 1122:1129 arg1 able 1200:1203 able 1200:1203 able 1200:1203 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 7 9 theme GnTI-dependent 1223:1236 arg1 defect 1252:1257 the GnTI-dependent glycosylation defect 1219:1257 the GnTI-dependent glycosylation defect 1219:1257 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 8 10 theme transmembrane 1319:1331 arg1 domain 1333:1338 the transmembrane domain 1315:1338 the transmembrane domain of GnTI 1315:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 8 10 theme transmembrane 1319:1331 arg1 GnTI 1343:1346 GnTI 1343:1346 GnTI 1343:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 3 11 theme proteins 374:381 arg1 series 355:360 a series 353:360 a series of chimeric proteins 353:381 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 6 12 theme stem 981:984 arg1 region 986:991 the stem region 977:991 the stem region of GnTI 977:999 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 5 13 theme GnTI 895:898 arg1 domain 885:890 the transmembrane domain 867:890 the transmembrane domain of GnTI 867:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 13 theme GnTI 895:898 arg1 determinant 913:923 the major determinant 903:923 the major determinant for its cis/medial-Golgi localization 903:961 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 13 theme GnTI 895:898 arg1 GnTI 895:898 GnTI 895:898 GnTI 895:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 7 14 with variant 1148:1154 arg1 localization 1179:1190 altered sub-Golgi localization 1161:1190 altered sub-Golgi localization 1161:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 3 15 theme Nicotiana 468:476 arg1 benthamiana 478:488 Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) 468:529 Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) 468:529 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 1 16 theme Golgi 204:208 arg1 stack 210:214 the Golgi stack 200:214 the Golgi stack 200:214 Golgi-resident type-II membrane proteins are asymmetrically distributed across the Golgi stack. 25230686 0 17 theme transmembrane 4:16 arg1 domain 18:23 The transmembrane domain 0:23 The transmembrane domain of N -acetylglucosaminyltransferase I 0:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 17 theme transmembrane 4:16 arg1 determinant 74:84 the key determinant 66:84 the key determinant for its Golgi subcompartmentation 66:118 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 17 theme transmembrane 4:16 arg1 -acetylglucosaminyltransferase 30:59 N -acetylglucosaminyltransferase I 28:61 N -acetylglucosaminyltransferase I 28:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 4 18 theme individual 623:632 arg1 domains 647:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 3 19 theme region 458:463 arg1 contribution 402:413 the contribution 398:413 the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi 398:616 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 3 20 theme cytoplasmic 422:432 arg1 region 458:463 the cytoplasmic, transmembrane and stem region 418:463 region 458:463 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 6 21 theme protein 1056:1062 arg1 formation 1072:1080 homomeric and heteromeric protein complex formation 1030:1080 homomeric and heteromeric protein complex formation 1030:1080 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 7 22 from able 1200:1203 arg1 thaliana 1122:1129 transgenic Arabidopsis thaliana 1099:1129 transgenic Arabidopsis thaliana 1099:1129 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 8 23 theme N-glycan 1463:1470 arg1 processing 1472:1481 efficient N-glycan processing 1453:1481 efficient N-glycan processing 1453:1481 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 4 24 theme trans-Golgi 700:710 arg1 α2,6-sialyltransferase 719:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase 685:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) 685:745 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 5 25 theme major 907:911 arg1 domain 885:890 the transmembrane domain 867:890 the transmembrane domain of GnTI 867:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 25 theme major 907:911 arg1 determinant 913:923 the major determinant 903:923 the major determinant for its cis/medial-Golgi localization 903:961 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 25 theme major 907:911 arg1 GnTI 895:898 GnTI 895:898 GnTI 895:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 4 26 theme GnTI 634:637 arg1 domains 647:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains 619:653 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 8 27 from distribution 1377:1388 arg1 cis/medial-Golgi 1397:1412 the cis/medial-Golgi 1393:1412 the cis/medial-Golgi in plants 1393:1422 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 5 28 theme co-localization 805:819 arg1 analysis 821:828 co-localization analysis 805:828 co-localization analysis 805:828 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 4 29 theme well-known 689:698 arg1 α2,6-sialyltransferase 719:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase 685:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) 685:745 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 0 30 theme -acetylglucosaminyltransferase 30:59 arg1 domain 18:23 The transmembrane domain 0:23 The transmembrane domain of N -acetylglucosaminyltransferase I 0:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 30 theme -acetylglucosaminyltransferase 30:59 arg1 determinant 74:84 the key determinant 66:84 the key determinant for its Golgi subcompartmentation 66:118 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 30 theme -acetylglucosaminyltransferase 30:59 arg1 -acetylglucosaminyltransferase 30:59 N -acetylglucosaminyltransferase I 28:61 N -acetylglucosaminyltransferase I 28:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 3 31 theme cis/medial-Golgi 539:554 arg1 localization 556:567 its cis/medial-Golgi localization 535:567 its cis/medial-Golgi localization 535:567 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 7 32 theme altered 1161:1167 arg1 localization 1179:1190 altered sub-Golgi localization 1161:1190 altered sub-Golgi localization 1161:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 4 33 dep α2,6-sialyltransferase 719:740 arg1 ST 743:744 ST 743:744 ST 743:744 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 3 34 theme N-acetylglucosaminyltransferase 490:520 arg1 I 522:522 N-acetylglucosaminyltransferase I 490:522 Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) 468:529 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 3 34 theme N-acetylglucosaminyltransferase 490:520 arg1 GnTI 525:528 GnTI 525:528 GnTI 525:528 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 0 35 theme N 28:28 arg1 -acetylglucosaminyltransferase 30:59 N -acetylglucosaminyltransferase I 28:61 N -acetylglucosaminyltransferase I 28:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 6 36 theme heteromeric 1044:1054 arg1 formation 1072:1080 homomeric and heteromeric protein complex formation 1030:1080 homomeric and heteromeric protein complex formation 1030:1080 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 3 37 theme stem 453:456 arg1 region 458:463 the cytoplasmic, transmembrane and stem region 418:463 region 458:463 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 4 38 theme enzyme 712:717 arg1 α2,6-sialyltransferase 719:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase 685:740 the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) 685:745 The individual GnTI protein domains were replaced with those from the well-known trans-Golgi enzyme α2,6-sialyltransferase (ST) and transiently expressed in Nicotiana benthamiana. 25230686 3 39 used used 348:351 arg2 we 345:346 we 345:346 we 345:346 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 8 40 theme sequence-specific 1285:1301 arg1 features 1303:1310 sequence-specific features 1285:1310 sequence-specific features in the transmembrane domain of GnTI 1285:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 8 41 theme GnTI 1343:1346 arg1 domain 1333:1338 the transmembrane domain 1315:1338 the transmembrane domain of GnTI 1315:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 8 41 theme GnTI 1343:1346 arg1 GnTI 1343:1346 GnTI 1343:1346 GnTI 1343:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 2 42 theme protein 247:253 arg1 unknown 330:336 unknown 330:336 unknown 330:336 The intrinsic features of the protein that determine its subcompartment-specific concentration are still largely unknown. 25230686 2 42 theme protein 247:253 arg1 features 231:238 The intrinsic features 217:238 The intrinsic features of the protein that determine its subcompartment-specific concentration 217:310 The intrinsic features of the protein that determine its subcompartment-specific concentration are still largely unknown. 25230686 1 43 theme Golgi-resident 121:134 arg1 proteins 153:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins are asymmetrically distributed across the Golgi stack. 25230686 3 44 theme transmembrane 435:447 arg1 region 458:463 the cytoplasmic, transmembrane and stem region 418:463 region 458:463 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 6 45 theme homomeric 1030:1038 arg1 formation 1072:1080 homomeric and heteromeric protein complex formation 1030:1080 homomeric and heteromeric protein complex formation 1030:1080 By contrast, the stem region of GnTI contributes predominately to homomeric and heteromeric protein complex formation. 25230686 3 46 dep benthamiana 478:488 arg1 I 522:522 N-acetylglucosaminyltransferase I 490:522 Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) 468:529 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 3 46 dep benthamiana 478:488 arg1 GnTI 525:528 GnTI 525:528 GnTI 525:528 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 7 47 theme Arabidopsis 1110:1120 arg1 thaliana 1122:1129 transgenic Arabidopsis thaliana 1099:1129 transgenic Arabidopsis thaliana 1099:1129 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 1 48 theme type-II 136:142 arg1 proteins 153:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins are asymmetrically distributed across the Golgi stack. 25230686 7 49 theme chimeric 1134:1141 arg1 variant 1148:1154 a chimeric GnTI variant 1132:1154 a chimeric GnTI variant with altered sub-Golgi localization 1132:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 7 49 theme chimeric 1134:1141 arg1 able 1200:1203 able 1200:1203 able 1200:1203 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 8 50 from features 1303:1310 arg1 domain 1333:1338 the transmembrane domain 1315:1338 the transmembrane domain of GnTI 1315:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 8 50 from features 1303:1310 arg1 GnTI 1343:1346 GnTI 1343:1346 GnTI 1343:1346 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 1 51 theme membrane 144:151 arg1 proteins 153:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins 121:160 Golgi-resident type-II membrane proteins are asymmetrically distributed across the Golgi stack. 25230686 5 52 theme N-glycan 834:841 arg1 profiling 843:851 N-glycan profiling 834:851 N-glycan profiling 834:851 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 7 53 theme GnTI 1143:1146 arg1 variant 1148:1154 a chimeric GnTI variant 1132:1154 a chimeric GnTI variant with altered sub-Golgi localization 1132:1190 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 7 53 theme GnTI 1143:1146 arg1 able 1200:1203 able 1200:1203 able 1200:1203 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 2 54 theme intrinsic 221:229 arg1 unknown 330:336 unknown 330:336 unknown 330:336 The intrinsic features of the protein that determine its subcompartment-specific concentration are still largely unknown. 25230686 2 54 theme intrinsic 221:229 arg1 features 231:238 The intrinsic features 217:238 The intrinsic features of the protein that determine its subcompartment-specific concentration 217:310 The intrinsic features of the protein that determine its subcompartment-specific concentration are still largely unknown. 25230686 0 55 theme key 70:72 arg1 determinant 74:84 the key determinant 66:84 the key determinant for its Golgi subcompartmentation 66:118 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 55 theme key 70:72 arg1 domain 18:23 The transmembrane domain 0:23 The transmembrane domain of N -acetylglucosaminyltransferase I 0:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 0 55 theme key 70:72 arg1 -acetylglucosaminyltransferase 30:59 N -acetylglucosaminyltransferase I 28:61 N -acetylglucosaminyltransferase I 28:61 The transmembrane domain of N -acetylglucosaminyltransferase I is the key determinant for its Golgi subcompartmentation. 25230686 5 56 theme cis/medial-Golgi 933:948 arg1 localization 950:961 its cis/medial-Golgi localization 929:961 its cis/medial-Golgi localization 929:961 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 2 57 theme subcompartment-specific 274:296 arg1 concentration 298:310 its subcompartment-specific concentration 270:310 its subcompartment-specific concentration 270:310 The intrinsic features of the protein that determine its subcompartment-specific concentration are still largely unknown. 25230686 7 58 theme transgenic 1099:1108 arg1 thaliana 1122:1129 transgenic Arabidopsis thaliana 1099:1129 transgenic Arabidopsis thaliana 1099:1129 Importantly, in transgenic Arabidopsis thaliana, a chimeric GnTI variant with altered sub-Golgi localization was not able to complement the GnTI-dependent glycosylation defect. 25230686 8 59 theme efficient 1453:1461 arg1 processing 1472:1481 efficient N-glycan processing 1453:1481 efficient N-glycan processing 1453:1481 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 3 60 theme benthamiana 478:488 arg1 region 458:463 the cytoplasmic, transmembrane and stem region 418:463 region 458:463 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 3 61 theme chimeric 365:372 arg1 proteins 374:381 chimeric proteins 365:381 chimeric proteins 365:381 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 8 62 theme steady-state 1364:1375 arg1 distribution 1377:1388 its steady-state distribution 1360:1388 its steady-state distribution in the cis/medial-Golgi in plants 1360:1422 Our results suggest that sequence-specific features in the transmembrane domain of GnTI account for its steady-state distribution in the cis/medial-Golgi in plants, which is a prerequisite for efficient N-glycan processing in vivo. 25230686 5 63 theme transmembrane 871:883 arg1 domain 885:890 the transmembrane domain 867:890 the transmembrane domain of GnTI 867:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 63 theme transmembrane 871:883 arg1 determinant 913:923 the major determinant 903:923 the major determinant for its cis/medial-Golgi localization 903:961 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 5 63 theme transmembrane 871:883 arg1 GnTI 895:898 GnTI 895:898 GnTI 895:898 Using co-localization analysis and N-glycan profiling, we show that the transmembrane domain of GnTI is the major determinant for its cis/medial-Golgi localization. 25230686 3 64 theme protein-protein 577:591 arg1 interaction 593:603 protein-protein interaction 577:603 protein-protein interaction in the Golgi 577:616 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 25230686 3 65 from interaction 593:603 arg1 Golgi 612:616 the Golgi 608:616 the Golgi 608:616 Here, we used a series of chimeric proteins to investigate the contribution of the cytoplasmic, transmembrane and stem region of Nicotiana benthamiana N-acetylglucosaminyltransferase I (GnTI) for its cis/medial-Golgi localization and for protein-protein interaction in the Golgi. 26956389 8 0 theme positive 1155:1162 arg1 mode 1191:1194 positive and negative fragmentation mode 1155:1194 positive and negative fragmentation mode 1155:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 10 1 theme human 1591:1595 arg1 virus 1614:1618 the human immunodeficiency virus 1587:1618 the human immunodeficiency virus 1587:1618 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 11 2 theme collision-induced 1690:1706 arg1 spectra 1721:1727 the negative ion collision-induced dissociation spectra 1673:1727 the negative ion collision-induced dissociation spectra of all resolved isomers 1673:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 8 3 theme negative 1168:1175 arg1 mode 1191:1194 positive and negative fragmentation mode 1155:1194 positive and negative fragmentation mode 1155:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 11 4 theme isomers 1745:1751 arg1 spectra 1721:1727 the negative ion collision-induced dissociation spectra 1673:1727 the negative ion collision-induced dissociation spectra of all resolved isomers 1673:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 3 5 with spectra 437:443 arg1 ions 474:477 mass-different fragment ions 450:477 mass-different fragment ions characterizing many of the isomers 450:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 10 6 theme separations 1444:1454 arg1 Examples 1425:1432 Examples 1425:1432 Examples of isomer separations 1425:1454 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 8 7 theme travelling-wave 1216:1230 arg1 spectrometry 1250:1261 travelling-wave ion mobility mass spectrometry 1216:1261 travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant 1216:1301 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 5 8 theme diagnostic 765:774 arg1 ions 785:788 the diagnostic fragment ions 761:788 the diagnostic fragment ions from the negative ion spectra 761:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 8 9 theme mobility 1236:1243 arg1 spectrometry 1250:1261 travelling-wave ion mobility mass spectrometry 1216:1261 travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant 1216:1301 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 4 10 from separation 522:531 arg1 modes 568:572 both ionization modes 552:572 both ionization modes 552:572 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 8 11 theme spectrometry 1250:1261 arg1 data 1263:1266 travelling-wave ion mobility mass spectrometry data 1216:1266 travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant 1216:1301 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 4 12 theme ion 536:538 arg1 mobility 540:547 ion mobility 536:547 ion mobility 536:547 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 5 13 theme fragment 744:751 arg1 ATDs 753:756 extracted fragment ATDs 734:756 extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra 734:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 2 14 theme mass 242:245 arg1 spectrometry 247:258 travelling-wave ion mobility mass spectrometry 213:258 travelling-wave ion mobility mass spectrometry 213:258 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 3 15 theme ion 334:336 arg1 fragmentation 338:350 Negative ion fragmentation 325:350 Negative ion fragmentation using collision-induced dissociation 325:387 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 5 16 from spectra 812:818 arg1 ATDs 753:756 extracted fragment ATDs 734:756 extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra 734:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 5 16 from spectra 812:818 arg1 ions 785:788 the diagnostic fragment ions 761:788 the diagnostic fragment ions from the negative ion spectra 761:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 2 17 theme high-mannose 285:296 arg1 N-glycans 298:306 high-mannose N-glycans 285:306 high-mannose N-glycans 285:306 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 0 18 theme high-mannose 63:74 arg1 N-glycans 76:84 high-mannose N-glycans 63:84 high-mannose N-glycans 63:84 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 9 19 theme complete 1309:1316 arg1 data 1342:1345 More complete collision cross section data 1304:1345 More complete collision cross section data 1304:1345 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 5 20 theme peak 929:932 arg1 areas 912:916 limited areas 904:916 limited areas of the ATD peak 904:932 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 9 21 theme cross 1328:1332 arg1 data 1342:1345 More complete collision cross section data 1304:1345 More complete collision cross section data 1304:1345 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 8 22 from sections 1128:1135 arg1 mode 1191:1194 positive and negative fragmentation mode 1155:1194 positive and negative fragmentation mode 1155:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 3 23 theme positive 424:431 arg1 spectra 437:443 positive ion spectra 424:443 positive ion spectra with mass-different fragment ions characterizing many of the isomers 424:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 10 24 theme chicken 1531:1537 arg1 ovalbumin 1539:1547 chicken ovalbumin 1531:1547 chicken ovalbumin 1531:1547 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 24 theme chicken 1531:1537 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 5 25 theme ion 808:810 arg1 spectra 812:818 the negative ion spectra 795:818 the negative ion spectra 795:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 0 26 theme ion 42:44 arg1 fragmentation 46:58 negative ion fragmentation 33:58 negative ion fragmentation 33:58 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 3 27 theme mass-different 450:463 arg1 ions 474:477 mass-different fragment ions 450:477 mass-different fragment ions characterizing many of the isomers 450:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 4 28 theme ionization 557:566 arg1 modes 568:572 both ionization modes 552:572 both ionization modes 552:572 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 10 29 theme well-characterized 1498:1515 arg1 gp120 1576:1580 gp120 1576:1580 gp120 1576:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 29 theme well-characterized 1498:1515 arg1 ovalbumin 1539:1547 chicken ovalbumin 1531:1547 chicken ovalbumin 1531:1547 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 29 theme well-characterized 1498:1515 arg1 thyroglobulin 1558:1570 porcine thyroglobulin 1550:1570 porcine thyroglobulin 1550:1570 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 29 theme well-characterized 1498:1515 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 0 30 theme Travelling-wave 0:14 arg1 mobility 20:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 9 31 theme negative 1364:1371 arg1 mode 1377:1380 negative ion mode 1364:1380 negative ion mode 1364:1380 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 10 32 attach released 1480:1487 arg1 gp120 1576:1580 gp120 1576:1580 gp120 1576:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 32 attach released 1480:1487 arg1 ovalbumin 1539:1547 chicken ovalbumin 1531:1547 chicken ovalbumin 1531:1547 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 32 attach released 1480:1487 arg1 thyroglobulin 1558:1570 porcine thyroglobulin 1550:1570 porcine thyroglobulin 1550:1570 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 32 attach released 1480:1487 arg2 N-glycans 1470:1478 N-glycans 1470:1478 N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus 1470:1618 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 32 attach released 1480:1487 arg1 virus 1614:1618 the human immunodeficiency virus 1587:1618 the human immunodeficiency virus 1587:1618 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 32 attach released 1480:1487 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 33 dep glycoproteins 1517:1529 arg1 gp120 1576:1580 gp120 1576:1580 gp120 1576:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 33 dep glycoproteins 1517:1529 arg1 ovalbumin 1539:1547 chicken ovalbumin 1531:1547 chicken ovalbumin 1531:1547 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 33 dep glycoproteins 1517:1529 arg1 thyroglobulin 1558:1570 porcine thyroglobulin 1550:1570 porcine thyroglobulin 1550:1570 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 33 dep glycoproteins 1517:1529 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 1 34 theme high-mannose 113:124 arg1 N-glycans 126:134 high-mannose N-glycans 113:134 high-mannose N-glycans 113:134 The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. 26956389 1 35 theme biological 161:170 arg1 events 184:189 biological recognition events 161:189 biological recognition events 161:189 The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. 26956389 6 36 located observed 971:978 arg1 cases 943:947 some cases 938:947 some cases 938:947 In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. 26956389 6 36 located observed 971:978 arg2 ATDs 961:964 asymmetric ATDs 950:964 asymmetric ATDs 950:964 In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. 26956389 0 37 theme negative 33:40 arg1 fragmentation 46:58 negative ion fragmentation 33:58 negative ion fragmentation 33:58 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 8 38 from isomers 1144:1150 arg1 mode 1191:1194 positive and negative fragmentation mode 1155:1194 positive and negative fragmentation mode 1155:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 9 39 theme diagnostic 1399:1408 arg1 ions 1419:1422 the diagnostic fragment ions 1395:1422 the diagnostic fragment ions 1395:1422 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 8 40 theme Collision 1112:1120 arg1 sections 1128:1135 Collision cross sections 1112:1135 Collision cross sections of the isomers in positive and negative fragmentation mode 1112:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 8 41 from mode 1191:1194 arg1 sections 1128:1135 Collision cross sections 1112:1135 Collision cross sections of the isomers in positive and negative fragmentation mode 1112:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 10 42 gly glycoproteins 1517:1529 arg1 gp120 1576:1580 gp120 1576:1580 gp120 1576:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 42 gly glycoproteins 1517:1529 arg1 ovalbumin 1539:1547 chicken ovalbumin 1531:1547 chicken ovalbumin 1531:1547 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 42 gly glycoproteins 1517:1529 arg1 thyroglobulin 1558:1570 porcine thyroglobulin 1550:1570 porcine thyroglobulin 1550:1570 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 42 gly glycoproteins 1517:1529 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 4 43 theme arrival 606:612 arg1 ATD 634:636 ATD 634:636 ATD 634:636 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 4 43 theme arrival 606:612 arg1 distributions 619:631 the arrival time distributions 602:631 the arrival time distributions (ATD) often showing little sign of isomers 602:674 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 2 44 theme ion 229:231 arg1 spectrometry 247:258 travelling-wave ion mobility mass spectrometry 213:258 travelling-wave ion mobility mass spectrometry 213:258 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 5 45 from ATDs 753:756 arg1 spectra 812:818 the negative ion spectra 795:818 the negative ion spectra 795:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 1 46 theme recognition 172:182 arg1 events 184:189 biological recognition events 161:189 biological recognition events 161:189 The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. 26956389 11 47 theme ion 1686:1688 arg1 spectra 1721:1727 the negative ion collision-induced dissociation spectra 1673:1727 the negative ion collision-induced dissociation spectra of all resolved isomers 1673:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 6 48 theme asymmetric 950:959 arg1 ATDs 961:964 asymmetric ATDs 950:964 asymmetric ATDs 950:964 In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. 26956389 10 49 theme immunodeficiency 1597:1612 arg1 virus 1614:1618 the human immunodeficiency virus 1587:1618 the human immunodeficiency virus 1587:1618 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 11 50 theme dissociation 1708:1719 arg1 spectra 1721:1727 the negative ion collision-induced dissociation spectra 1673:1727 the negative ion collision-induced dissociation spectra of all resolved isomers 1673:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 4 51 theme little 653:658 arg1 sign 660:663 little sign 653:663 little sign of isomers 653:674 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 5 52 theme limited 904:910 arg1 areas 912:916 limited areas 904:916 limited areas of the ATD peak 904:932 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 8 53 theme fragmentation 1177:1189 arg1 mode 1191:1194 positive and negative fragmentation mode 1155:1194 positive and negative fragmentation mode 1155:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 11 54 theme resolved 1736:1743 arg1 isomers 1745:1751 all resolved isomers 1732:1751 all resolved isomers 1732:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 8 55 theme dextran 1274:1280 arg1 glycans 1282:1288 dextran glycans 1274:1288 dextran glycans 1274:1288 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 10 56 theme isomer 1437:1442 arg1 separations 1444:1454 isomer separations 1437:1454 isomer separations 1437:1454 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 3 57 theme Negative 325:332 arg1 fragmentation 338:350 Negative ion fragmentation 325:350 Negative ion fragmentation using collision-induced dissociation 325:387 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 5 58 theme fragment 776:783 arg1 ions 785:788 the diagnostic fragment ions 761:788 the diagnostic fragment ions from the negative ion spectra 761:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 2 59 theme isomer 264:269 arg1 separation 271:280 isomer separation 264:280 isomer separation of high-mannose N-glycans 264:306 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 8 60 theme ion 1232:1234 arg1 spectrometry 1250:1261 travelling-wave ion mobility mass spectrometry 1216:1261 travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant 1216:1301 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 5 61 theme extracted 734:742 arg1 ATDs 753:756 extracted fragment ATDs 734:756 extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra 734:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 2 62 theme spectrometry 247:258 arg1 utility 202:208 the utility 198:208 the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans 198:306 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 1 63 theme N-glycans 126:134 arg1 structure 100:108 The isomeric structure 87:108 The isomeric structure of high-mannose N-glycans 87:134 The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. 26956389 8 64 theme mass 1245:1248 arg1 spectrometry 1250:1261 travelling-wave ion mobility mass spectrometry 1216:1261 travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant 1216:1301 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 3 65 theme collision-induced 358:374 arg1 dissociation 376:387 collision-induced dissociation 358:387 collision-induced dissociation 358:387 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 2 66 theme N-glycans 298:306 arg1 separation 271:280 isomer separation 264:280 isomer separation of high-mannose N-glycans 264:306 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 4 67 theme Isomer 515:520 arg1 separation 522:531 Isomer separation 515:531 Isomer separation by ion mobility in both ionization modes 515:572 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 0 68 theme N-glycans 76:84 arg1 mobility 20:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 0 68 theme N-glycans 76:84 arg1 fragmentation 46:58 negative ion fragmentation 33:58 negative ion fragmentation 33:58 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26956389 5 69 theme ATD 925:927 arg1 peak 929:932 the ATD peak 921:932 the ATD peak 921:932 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 3 70 theme informative 399:409 arg1 spectra 411:417 more informative spectra 394:417 more informative spectra 394:417 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 9 71 theme collision 1318:1326 arg1 data 1342:1345 More complete collision cross section data 1304:1345 More complete collision cross section data 1304:1345 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 5 72 from areas 912:916 arg1 ions 894:897 ions 894:897 ions from limited areas of the ATD peak 894:932 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 9 73 theme section 1334:1340 arg1 data 1342:1345 More complete collision cross section data 1304:1345 More complete collision cross section data 1304:1345 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 3 74 theme ion 433:435 arg1 spectra 437:443 positive ion spectra 424:443 positive ion spectra with mass-different fragment ions characterizing many of the isomers 424:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 5 75 theme negative 799:806 arg1 spectra 812:818 the negative ion spectra 795:818 the negative ion spectra 795:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 5 76 theme isomers 858:864 arg1 spectra 843:849 the fragmentation spectra 825:849 the fragmentation spectra of the isomers 825:864 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 1 77 theme isomeric 91:98 arg1 structure 100:108 The isomeric structure 87:108 The isomeric structure of high-mannose N-glycans 87:134 The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. 26956389 3 78 theme fragment 465:472 arg1 ions 474:477 mass-different fragment ions 450:477 mass-different fragment ions characterizing many of the isomers 450:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 3 79 theme isomers 506:512 arg1 many 494:497 many 494:497 many 494:497 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 3 79 theme isomers 506:512 arg1 isomers 506:512 the isomers 502:512 the isomers 502:512 Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. 26956389 9 80 theme ion 1373:1375 arg1 mode 1377:1380 negative ion mode 1364:1380 negative ion mode 1364:1380 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 10 81 theme porcine 1550:1556 arg1 thyroglobulin 1558:1570 porcine thyroglobulin 1550:1570 porcine thyroglobulin 1550:1570 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 10 81 theme porcine 1550:1556 arg1 glycoproteins 1517:1529 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 1494:1580 Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. 26956389 5 82 theme fragmentation 829:841 arg1 spectra 843:849 the fragmentation spectra 825:849 the fragmentation spectra of the isomers 825:864 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 8 83 theme cross 1122:1126 arg1 sections 1128:1135 Collision cross sections 1112:1135 Collision cross sections of the isomers in positive and negative fragmentation mode 1112:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 5 84 theme ions 785:788 arg1 ATDs 753:756 extracted fragment ATDs 734:756 extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra 734:818 However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. 26956389 4 85 theme time 614:617 arg1 ATD 634:636 ATD 634:636 ATD 634:636 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 4 85 theme time 614:617 arg1 distributions 619:631 the arrival time distributions 602:631 the arrival time distributions (ATD) often showing little sign of isomers 602:674 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 2 86 theme mobility 233:240 arg1 spectrometry 247:258 travelling-wave ion mobility mass spectrometry 213:258 travelling-wave ion mobility mass spectrometry 213:258 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 11 87 theme cross-sectional 1640:1654 arg1 data 1656:1659 the cross-sectional data 1636:1659 the cross-sectional data 1636:1659 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 9 88 theme fragment 1410:1417 arg1 ions 1419:1422 the diagnostic fragment ions 1395:1422 the diagnostic fragment ions 1395:1422 More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. 26956389 4 89 theme isomers 668:674 arg1 sign 660:663 little sign 653:663 little sign of isomers 653:674 Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. 26956389 2 90 theme travelling-wave 213:227 arg1 spectrometry 247:258 travelling-wave ion mobility mass spectrometry 213:258 travelling-wave ion mobility mass spectrometry 213:258 Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. 26956389 11 91 theme negative 1677:1684 arg1 spectra 1721:1727 the negative ion collision-induced dissociation spectra 1673:1727 the negative ion collision-induced dissociation spectra of all resolved isomers 1673:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 8 92 theme isomers 1144:1150 arg1 sections 1128:1135 Collision cross sections 1112:1135 Collision cross sections of the isomers in positive and negative fragmentation mode 1112:1194 Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. 26956389 11 93 theme spectra 1721:1727 arg1 details 1662:1668 details 1662:1668 details of the negative ion collision-induced dissociation spectra of all resolved isomers 1662:1751 In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed. 26956389 0 94 theme ion 16:18 arg1 mobility 20:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility 0:27 Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans. 26749838 0 0 theme Convolvulus 73:83 arg1 althaeoides 85:95 Convolvulus althaeoides 73:95 Convolvulus althaeoides 73:95 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 4 1 theme novel 674:678 arg1 This 664:667 This 664:667 This 664:667 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 4 1 theme novel 674:678 arg1 trisaccharide 680:692 a novel trisaccharide 672:692 a novel trisaccharide within the flavonoids 672:714 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 6 2 theme many 1002:1005 arg1 anthocyanins 1015:1026 many complex anthocyanins 1002:1026 many complex anthocyanins 1002:1026 Although many complex anthocyanins have been found amongst the Convolvulaceae, the genus Convolvulus had not yet been surveyed for anthocyanins and thus this account could be of significance within the current chemotaxonomy of this family. 26749838 1 3 attach isolated 135:142 arg1 flowers 154:160 pink flowers 149:160 pink flowers of Convolvulus althaeoides 149:187 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 1 3 attach isolated 135:142 arg2 trioside 122:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 2 4 dep 4-O- 383:386 arg1 -β-D-glucopyranosyl 405:423 -β-D-glucopyranosyl 405:423 -β-D-glucopyranosyl 405:423 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 6 5 theme family 1225:1230 arg1 chemotaxonomy 1203:1215 the current chemotaxonomy 1191:1215 the current chemotaxonomy of this family 1191:1230 Although many complex anthocyanins have been found amongst the Convolvulaceae, the genus Convolvulus had not yet been surveyed for anthocyanins and thus this account could be of significance within the current chemotaxonomy of this family. 26749838 5 6 theme rhamnose 965:972 arg1 anthocyanins 934:945 anthocyanins 934:945 anthocyanins of a glycosylated rhamnose 934:972 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 2 7 theme 1H 280:281 arg1 NMR 283:285 1H NMR 280:285 1H NMR 280:285 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 7 theme 1H 280:281 arg1 COSY 294:297 1H-1H COSY 288:297 1H-1H COSY 288:297 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 0 8 theme althaeoides 85:95 arg1 Flowers 62:68 Flowers 62:68 Flowers of Convolvulus althaeoides 62:95 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 4 9 theme complete 731:738 arg1 characterization 740:755 the first complete characterization 721:755 the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins 721:844 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 3 10 theme absolute 497:504 arg1 configuration 506:518 The absolute configuration 493:518 The absolute configuration of the aldose enantiomer moieties 493:552 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 5 11 theme rhamnose 867:874 arg1 moieties 876:883 rhamnose moieties 867:883 rhamnose moieties 867:883 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 5 12 gly glycosylated 952:963 arg1 rhamnose 965:972 a glycosylated rhamnose 950:972 a glycosylated rhamnose 950:972 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 1 13 theme Convolvulus 165:175 arg1 althaeoides 177:187 Convolvulus althaeoides 165:187 Convolvulus althaeoides 165:187 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 2 14 theme 4-O- 383:386 arg1 3-O-[6-O- 373:381 cyanidin 3-O-[6-O- 364:381 cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside 364:490 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 14 theme 4-O- 383:386 arg1 -β-L-rhamnopyranosyl 425:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 5 15 from occurrence 920:929 arg1 anthocyanins 934:945 anthocyanins 934:945 anthocyanins of a glycosylated rhamnose 934:972 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 5 16 theme unique 979:984 arg1 trait 986:990 a unique trait 977:990 a unique trait 977:990 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 5 16 theme unique 979:984 arg1 occurrence 920:929 the occurrence 916:929 the occurrence in anthocyanins of a glycosylated rhamnose 916:972 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 1 17 theme althaeoides 177:187 arg1 flowers 154:160 pink flowers 149:160 pink flowers of Convolvulus althaeoides 149:187 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 0 18 theme Novel 2:6 arg1 Anthocyanin 17:27 A Novel Acylated Anthocyanin 0:27 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 0:96 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 2 19 theme 1H-1H 288:292 arg1 CAPT 326:329 13C CAPT 322:329 13C CAPT 322:329 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 19 theme 1H-1H 288:292 arg1 NMR 283:285 1H NMR 280:285 1H NMR 280:285 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 19 theme 1H-1H 288:292 arg1 HSQC 310:313 HSQC 310:313 HSQC 310:313 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 19 theme 1H-1H 288:292 arg1 HMBC 316:319 HMBC 316:319 HMBC 316:319 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 19 theme 1H-1H 288:292 arg1 COSY 294:297 1H-1H COSY 288:297 1H-1H COSY 288:297 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 19 theme 1H-1H 288:292 arg1 TOCSY 303:307 1D TOCSY 300:307 1D TOCSY 300:307 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 20 theme 13C 322:324 arg1 CAPT 326:329 13C CAPT 322:329 13C CAPT 322:329 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 20 theme 13C 322:324 arg1 COSY 294:297 1H-1H COSY 288:297 1H-1H COSY 288:297 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 5 21 from acylation 854:862 arg1 position 888:895 position 4 888:897 position 4 888:897 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 3 22 theme aldose 527:532 arg1 moieties 545:552 the aldose enantiomer moieties 523:552 the aldose enantiomer moieties 523:552 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 2 23 theme cyanidin 364:371 arg1 3-O-[6-O- 373:381 cyanidin 3-O-[6-O- 364:381 cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside 364:490 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 23 theme cyanidin 364:371 arg1 structure 336:344 its structure 332:344 its structure 332:344 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 23 theme cyanidin 364:371 arg1 -β-L-rhamnopyranosyl 425:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 6 24 theme genus 1076:1080 arg1 Convolvulus 1082:1092 the genus Convolvulus 1072:1092 the genus Convolvulus 1072:1092 Although many complex anthocyanins have been found amongst the Convolvulaceae, the genus Convolvulus had not yet been surveyed for anthocyanins and thus this account could be of significance within the current chemotaxonomy of this family. 26749838 4 25 theme first 725:729 arg1 characterization 740:755 the first complete characterization 721:755 the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins 721:844 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 0 26 theme Acylated 8:15 arg1 Anthocyanin 17:27 A Novel Acylated Anthocyanin 0:27 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 0:96 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 3 27 theme thiazolidine 610:621 arg1 derivatives 639:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 1 28 theme acylated 101:108 arg1 trioside 122:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 3 29 theme diastereoisomer 623:637 arg1 derivatives 639:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 1 30 theme chromatographic 212:226 arg1 techniques 228:237 chromatographic techniques 212:237 chromatographic techniques 212:237 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 2 31 dep MS 256:257 arg1 the 243:245 the 243:245 the 243:245 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 31 dep MS 256:257 arg1 basis 247:251 basis 247:251 basis 247:251 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 32 theme -5-O-β-D-glucopyranoside 467:490 arg1 3-O-[6-O- 373:381 cyanidin 3-O-[6-O- 364:381 cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside 364:490 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 32 theme -5-O-β-D-glucopyranoside 467:490 arg1 structure 336:344 its structure 332:344 its structure 332:344 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 32 theme -5-O-β-D-glucopyranoside 467:490 arg1 -β-L-rhamnopyranosyl 425:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl 383:444 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 0 33 from Flowers 62:68 arg1 Trisaccharide 43:55 a Linear Trisaccharide 34:55 a Linear Trisaccharide from Flowers of Convolvulus althaeoides 34:95 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 0 34 with Anthocyanin 17:27 arg1 Trisaccharide 43:55 a Linear Trisaccharide 34:55 a Linear Trisaccharide from Flowers of Convolvulus althaeoides 34:95 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 1 35 theme techniques 228:237 arg1 combination 197:207 a combination 195:207 a combination of chromatographic techniques 195:237 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 3 36 theme corresponding 596:608 arg1 derivatives 639:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 the corresponding thiazolidine diastereoisomer derivatives 592:649 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 3 37 theme enantiomer 534:543 arg1 moieties 545:552 the aldose enantiomer moieties 523:552 the aldose enantiomer moieties 523:552 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 1 38 theme anthocyanin 110:120 arg1 trioside 122:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside 98:129 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 0 39 theme Linear 36:41 arg1 Trisaccharide 43:55 a Linear Trisaccharide 34:55 a Linear Trisaccharide from Flowers of Convolvulus althaeoides 34:95 A Novel Acylated Anthocyanin with a Linear Trisaccharide from Flowers of Convolvulus althaeoides. 26749838 3 40 theme moieties 545:552 arg1 configuration 506:518 The absolute configuration 493:518 The absolute configuration of the aldose enantiomer moieties 493:552 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 2 41 theme 1D 300:301 arg1 COSY 294:297 1H-1H COSY 288:297 1H-1H COSY 288:297 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 2 41 theme 1D 300:301 arg1 TOCSY 303:307 1D TOCSY 300:307 1D TOCSY 300:307 On the basis of MS (MALDI-TOF) and NMR (1H NMR, 1H-1H COSY, 1D TOCSY, HSQC, HMBC, 13C CAPT) its structure was determined as cyanidin 3-O-[6-O-(4-O- (6-O-(E-caffeoyl)-β-D-glucopyranosyl)-β-L-rhamnopyranosyl)-β-D-glucopyranoside]-5-O-β-D-glucopyranoside. 26749838 4 42 theme glucosyl-rhamnosyl-glucoside 769:796 arg1 characterization 740:755 the first complete characterization 721:755 the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins 721:844 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 4 42 theme glucosyl-rhamnosyl-glucoside 769:796 arg1 This 664:667 This 664:667 This 664:667 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 4 42 theme glucosyl-rhamnosyl-glucoside 769:796 arg1 trisaccharide 680:692 a novel trisaccharide 672:692 a novel trisaccharide within the flavonoids 672:714 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 3 43 theme derivatives 639:649 arg1 separation 578:587 the separation 574:587 the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD 574:661 The absolute configuration of the aldose enantiomer moieties was determined from the separation of the corresponding thiazolidine diastereoisomer derivatives by HPLC-DAD. 26749838 4 44 theme linear 762:767 arg1 4'-glucosylrutinoside 799:819 4'-glucosylrutinoside 799:819 4'-glucosylrutinoside 799:819 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 4 44 theme linear 762:767 arg1 glucosyl-rhamnosyl-glucoside 769:796 a linear glucosyl-rhamnosyl-glucoside 760:796 a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins 760:844 This is a novel trisaccharide within the flavonoids, and the first complete characterization of a linear glucosyl-rhamnosyl-glucoside (4'-glucosylrutinoside) within the anthocyanins. 26749838 6 45 theme current 1195:1201 arg1 chemotaxonomy 1203:1215 the current chemotaxonomy 1191:1215 the current chemotaxonomy of this family 1191:1230 Although many complex anthocyanins have been found amongst the Convolvulaceae, the genus Convolvulus had not yet been surveyed for anthocyanins and thus this account could be of significance within the current chemotaxonomy of this family. 26749838 5 46 theme glycosylated 952:963 arg1 rhamnose 965:972 a glycosylated rhamnose 950:972 a glycosylated rhamnose 950:972 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 5 47 theme moieties 876:883 arg1 acylation 854:862 acylation 854:862 acylation of rhamnose moieties in position 4 854:897 Whilst acylation of rhamnose moieties in position 4 is quite common, the occurrence in anthocyanins of a glycosylated rhamnose is a unique trait. 26749838 1 48 theme pink 149:152 arg1 flowers 154:160 pink flowers 149:160 pink flowers of Convolvulus althaeoides 149:187 An acylated anthocyanin trioside was isolated from pink flowers of Convolvulus althaeoides using a combination of chromatographic techniques. 26749838 6 49 theme complex 1007:1013 arg1 anthocyanins 1015:1026 many complex anthocyanins 1002:1026 many complex anthocyanins 1002:1026 Although many complex anthocyanins have been found amongst the Convolvulaceae, the genus Convolvulus had not yet been surveyed for anthocyanins and thus this account could be of significance within the current chemotaxonomy of this family. 28052259 2 0 theme crystal 339:345 arg1 structures 347:356 multiple crystal structures 330:356 multiple crystal structures 330:356 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 3 1 theme third 772:776 arg1 modules 778:784 the second and the third modules 753:784 the second and the third modules 753:784 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 2 2 theme molecular 450:458 arg1 architecture 460:471 the overall molecular architecture 438:471 the overall molecular architecture of the full-length LRP6 438:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 2 3 theme multiple 330:337 arg1 structures 347:356 multiple crystal structures 330:356 multiple crystal structures 330:356 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 5 4 theme ectodomain 1060:1069 arg1 change 1086:1091 the ectodomain conformational change 1056:1091 the ectodomain conformational change 1056:1091 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 5 5 theme receptor 1114:1121 arg1 oligomerization 1123:1137 receptor oligomerization 1114:1137 receptor oligomerization 1114:1137 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 5 6 theme Wnt 1003:1005 arg1 Dkk1 1018:1021 the Wnt antagonist Dkk1 999:1021 the Wnt antagonist Dkk1 999:1021 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 1 7 theme higher 243:248 arg1 platform 266:273 a higher order signaling platform 241:273 a higher order signaling platform 241:273 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 3 8 theme large 701:705 arg1 motion 715:720 a large bending motion 699:720 a large bending motion hinged at the junction between the second and the third modules 699:784 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 1 9 from ligands 292:298 arg1 surface 312:318 the cell surface 303:318 the cell surface 303:318 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 0 10 gly N-glycosylation 62:76 arg1 Dkk1 116:119 the Wnt Antagonist Dkk1 97:119 the Wnt Antagonist Dkk1 97:119 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 1 11 theme order 250:254 arg1 platform 266:273 a higher order signaling platform 241:273 a higher order signaling platform 241:273 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 2 12 theme LRP6 492:495 arg1 architecture 460:471 the overall molecular architecture 438:471 the overall molecular architecture of the full-length LRP6 438:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 2 13 from view 430:433 arg1 architecture 460:471 the overall molecular architecture 438:471 the overall molecular architecture of the full-length LRP6 438:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 1 14 gly glycoprotein 186:197 arg1 glycoprotein 186:197 a single-pass membrane glycoprotein 163:197 a single-pass membrane glycoprotein with a large modular ectodomain 163:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 14 gly glycoprotein 186:197 arg1 protein 143:149 LDL-receptor-related protein 6 122:151 LDL-receptor-related protein 6 (LRP6) 122:158 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 3 15 from states 595:600 arg1 solution 638:645 solution 638:645 solution 638:645 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 2 16 theme full-length 480:490 arg1 LRP6 492:495 the full-length LRP6 476:495 the full-length LRP6 476:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 2 17 theme overall 442:448 arg1 architecture 460:471 the overall molecular architecture 438:471 the overall molecular architecture of the full-length LRP6 438:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 5 18 theme conformational 1071:1084 arg1 change 1086:1091 the ectodomain conformational change 1056:1091 the ectodomain conformational change 1056:1091 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 0 19 theme Wnt 101:103 arg1 Dkk1 116:119 the Wnt Antagonist Dkk1 97:119 the Wnt Antagonist Dkk1 97:119 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 2 20 theme consensus 420:428 arg1 view 430:433 a consensus view 418:433 a consensus view on the overall molecular architecture of the full-length LRP6 418:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 4 21 theme motion 827:832 arg1 extent 804:809 the extent 800:809 the extent of inter-domain motion 800:832 Importantly, the extent of inter-domain motion is modulated by evolutionarily conserved N-glycan chains proximal to the joint. 28052259 3 22 theme bending 707:713 arg1 motion 715:720 a large bending motion 699:720 a large bending motion hinged at the junction between the second and the third modules 699:784 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 0 23 theme Conformational 0:13 arg1 Freedom 15:21 Conformational Freedom 0:21 Conformational Freedom of the LRP6 Ectodomain 0:44 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 1 24 theme signaling 256:264 arg1 platform 266:273 a higher order signaling platform 241:273 a higher order signaling platform 241:273 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 4 25 theme inter-domain 814:825 arg1 motion 827:832 inter-domain motion 814:832 inter-domain motion 814:832 Importantly, the extent of inter-domain motion is modulated by evolutionarily conserved N-glycan chains proximal to the joint. 28052259 3 26 theme negative-stain 536:549 arg1 microscopy 560:569 negative-stain electron microscopy 536:569 negative-stain electron microscopy 536:569 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 1 27 theme single-pass 165:175 arg1 glycoprotein 186:197 a single-pass membrane glycoprotein 163:197 a single-pass membrane glycoprotein with a large modular ectodomain 163:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 27 theme single-pass 165:175 arg1 protein 143:149 LDL-receptor-related protein 6 122:151 LDL-receptor-related protein 6 (LRP6) 122:158 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 0 28 theme Dkk1 116:119 arg1 N-glycosylation 62:76 N-glycosylation 62:76 N-glycosylation 62:76 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 0 28 theme Dkk1 116:119 arg1 Binding 86:92 the Binding 82:92 the Binding of the Wnt Antagonist Dkk1 82:119 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 5 29 theme LRP6 937:940 arg1 ectodomain 942:951 the LRP6 ectodomain 933:951 the LRP6 ectodomain 933:951 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 1 30 theme membrane 177:184 arg1 glycoprotein 186:197 a single-pass membrane glycoprotein 163:197 a single-pass membrane glycoprotein with a large modular ectodomain 163:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 30 theme membrane 177:184 arg1 protein 143:149 LDL-receptor-related protein 6 122:151 LDL-receptor-related protein 6 (LRP6) 122:158 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 0 31 theme Antagonist 105:114 arg1 Dkk1 116:119 the Wnt Antagonist Dkk1 97:119 the Wnt Antagonist Dkk1 97:119 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 5 32 theme potential 1037:1045 arg1 role 1047:1050 a potential role 1035:1050 a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling 1035:1151 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 2 33 theme ectodomain 398:407 arg1 fragments 376:384 fragments 376:384 fragments of the LRP6 ectodomain 376:407 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 0 34 theme Ectodomain 35:44 arg1 Freedom 15:21 Conformational Freedom 0:21 Conformational Freedom of the LRP6 Ectodomain 0:44 Conformational Freedom of the LRP6 Ectodomain Is Regulated by N-glycosylation and the Binding of the Wnt Antagonist Dkk1. 28052259 3 35 theme LRP6 630:633 arg1 LRP6 630:633 LRP6 630:633 LRP6 630:633 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 3 35 theme LRP6 630:633 arg1 ectodomain 616:625 the entire ectodomain 605:625 the entire ectodomain of LRP6 605:633 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 1 36 theme Wnt 288:290 arg1 ligands 292:298 Wnt ligands 288:298 Wnt ligands on the cell surface 288:318 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 37 with glycoprotein 186:197 arg1 ectodomain 220:229 a large modular ectodomain 204:229 a large modular ectodomain 204:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 3 38 theme entire 609:614 arg1 LRP6 630:633 LRP6 630:633 LRP6 630:633 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 3 38 theme entire 609:614 arg1 ectodomain 616:625 the entire ectodomain 605:625 the entire ectodomain of LRP6 605:633 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 5 39 from role 1047:1050 arg1 regulation 1100:1109 the regulation 1096:1109 the regulation of receptor oligomerization and signaling 1096:1151 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 3 40 used used 531:534 arg2 we 528:529 we 528:529 we 528:529 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 3 41 theme second 757:762 arg1 modules 778:784 the second and the third modules 753:784 the second and the third modules 753:784 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 4 42 theme N-glycan 875:882 arg1 chains 884:889 evolutionarily conserved N-glycan chains 850:889 evolutionarily conserved N-glycan chains proximal to the joint 850:911 Importantly, the extent of inter-domain motion is modulated by evolutionarily conserved N-glycan chains proximal to the joint. 28052259 3 43 theme ectodomain 616:625 arg1 states 595:600 conformational states 580:600 conformational states of the entire ectodomain of LRP6 in solution 580:645 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 3 44 theme electron 551:558 arg1 microscopy 560:569 negative-stain electron microscopy 536:569 negative-stain electron microscopy 536:569 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 2 45 theme dynamic 505:511 arg1 aspects 513:519 its dynamic aspects 501:519 its dynamic aspects 501:519 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 28052259 5 46 theme signaling 1143:1151 arg1 regulation 1100:1109 the regulation 1096:1109 the regulation of receptor oligomerization and signaling 1096:1151 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 3 47 theme four-module 666:676 arg1 ectodomain 678:687 the four-module ectodomain 662:687 the four-module ectodomain 662:687 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 5 48 theme antagonist 1007:1016 arg1 Dkk1 1018:1021 the Wnt antagonist Dkk1 999:1021 the Wnt antagonist Dkk1 999:1021 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 5 49 theme oligomerization 1123:1137 arg1 regulation 1100:1109 the regulation 1096:1109 the regulation of receptor oligomerization and signaling 1096:1151 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 4 50 theme proximal 891:898 arg1 chains 884:889 evolutionarily conserved N-glycan chains 850:889 evolutionarily conserved N-glycan chains proximal to the joint 850:911 Importantly, the extent of inter-domain motion is modulated by evolutionarily conserved N-glycan chains proximal to the joint. 28052259 1 51 theme large 206:210 arg1 ectodomain 220:229 a large modular ectodomain 204:229 a large modular ectodomain 204:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 52 theme LDL-receptor-related 122:141 arg1 glycoprotein 186:197 a single-pass membrane glycoprotein 163:197 a single-pass membrane glycoprotein with a large modular ectodomain 163:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 52 theme LDL-receptor-related 122:141 arg1 protein 143:149 LDL-receptor-related protein 6 122:151 LDL-receptor-related protein 6 (LRP6) 122:158 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 52 theme LDL-receptor-related 122:141 arg1 LRP6 154:157 LRP6 154:157 LRP6 154:157 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 1 53 theme modular 212:218 arg1 ectodomain 220:229 a large modular ectodomain 204:229 a large modular ectodomain 204:229 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 3 54 theme conformational 580:593 arg1 states 595:600 conformational states 580:600 conformational states of the entire ectodomain of LRP6 in solution 580:645 Here, we used negative-stain electron microscopy to probe conformational states of the entire ectodomain of LRP6 in solution and found that the four-module ectodomain undergoes a large bending motion hinged at the junction between the second and the third modules. 28052259 4 55 theme conserved 865:873 arg1 chains 884:889 evolutionarily conserved N-glycan chains 850:889 evolutionarily conserved N-glycan chains proximal to the joint 850:911 Importantly, the extent of inter-domain motion is modulated by evolutionarily conserved N-glycan chains proximal to the joint. 28052259 5 56 with complexation 981:992 arg1 Dkk1 1018:1021 the Wnt antagonist Dkk1 999:1021 the Wnt antagonist Dkk1 999:1021 We also found that the LRP6 ectodomain becomes highly compact upon complexation with the Wnt antagonist Dkk1, suggesting a potential role for the ectodomain conformational change in the regulation of receptor oligomerization and signaling. 28052259 1 57 theme cell 307:310 arg1 surface 312:318 the cell surface 303:318 the cell surface 303:318 LDL-receptor-related protein 6 (LRP6) is a single-pass membrane glycoprotein with a large modular ectodomain and forms a higher order signaling platform upon binding Wnt ligands on the cell surface. 28052259 2 58 from aspects 513:519 arg1 architecture 460:471 the overall molecular architecture 438:471 the overall molecular architecture of the full-length LRP6 438:495 Although multiple crystal structures are available for fragments of the LRP6 ectodomain, we lack a consensus view on the overall molecular architecture of the full-length LRP6 and its dynamic aspects. 26185906 8 0 from N-glycans 1552:1560 arg1 non-attractors 1565:1578 non-attractors 1565:1578 non-attractors 1565:1578 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 8 0 from N-glycans 1552:1560 arg1 galactose 1593:1601 terminal galactose 1584:1601 terminal galactose 1584:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 9 1 theme first 1677:1681 arg1 evidence 1683:1690 the first evidence 1673:1690 the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs 1673:1762 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 7 2 theme type 1404:1407 arg1 N-glycans 1409:1417 increased high mannose type N-glycans 1381:1417 increased high mannose type N-glycans 1381:1417 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 1 3 theme marrow-derived 86:99 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 3 theme marrow-derived 86:99 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 7 4 theme high 1391:1394 arg1 N-glycans 1409:1417 increased high mannose type N-glycans 1381:1417 increased high mannose type N-glycans 1381:1417 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 7 5 with validation 1240:1249 arg1 deglycosylation 1273:1287 topical PNGase F deglycosylation 1256:1287 topical PNGase F deglycosylation 1256:1287 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 9 6 theme glycomic 1709:1716 arg1 profiles 1718:1725 differential glycomic profiles 1696:1725 differential glycomic profiles in attractor and non-attractor GSCXs 1696:1762 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 7 7 from validation 1240:1249 arg1 regions 1302:1308 the tumor regions 1292:1308 the tumor regions of xenograft tissue 1292:1328 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 8 from non-attractors 1565:1578 arg1 prevalence 1506:1515 the prevalence 1502:1515 the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors 1502:1650 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 2 9 theme BM-hMSCs 276:283 arg1 property 264:271 this unique property 252:271 this unique property of BM-hMSCs 252:283 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 26185906 9 10 from profiles 1718:1725 arg1 non-attractor 1744:1756 non-attractor 1744:1756 non-attractor 1744:1756 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 9 10 from profiles 1718:1725 arg1 attractor 1730:1738 attractor 1730:1738 attractor 1730:1738 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 7 11 attach presence 1369:1376 arg2 N-glycans 1409:1417 increased high mannose type N-glycans 1381:1417 increased high mannose type N-glycans 1381:1417 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 7 11 attach presence 1369:1376 arg1 non-attractors 1426:1439 the non-attractors 1422:1439 the non-attractors 1422:1439 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 12 from N-glycans 1628:1636 arg1 non-attractors 1565:1578 non-attractors 1565:1578 non-attractors 1565:1578 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 8 12 from N-glycans 1628:1636 arg1 galactose 1593:1601 terminal galactose 1584:1601 terminal galactose 1584:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 5 13 theme BM-hMSCs 1002:1009 arg1 engraftment 987:997 the engraftment 983:997 the engraftment of BM-hMSCs in the tumor 983:1022 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 3 14 theme glycan-protein 573:586 arg1 adhesion 588:595 glycan-glycan and glycan-protein adhesion 555:595 glycan-glycan and glycan-protein adhesion between the cell and endothelium 555:628 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 3 15 dep cell 609:612 arg1 the 605:607 the 605:607 the 605:607 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 6 16 theme N-glycan 1175:1182 arg1 biosynthesis 1184:1195 high mannose type N-glycan biosynthesis 1157:1195 high mannose type N-glycan biosynthesis 1157:1195 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 4 17 theme xenograft 702:710 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 3 18 theme glycan-glycan 555:567 arg1 adhesion 588:595 glycan-glycan and glycan-protein adhesion 555:595 glycan-glycan and glycan-protein adhesion between the cell and endothelium 555:628 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 8 19 theme Additional 1442:1451 arg1 evidence 1453:1460 Additional evidence 1442:1460 Additional evidence provided by our glycomic study 1442:1491 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 6 20 theme mannose 1162:1168 arg1 biosynthesis 1184:1195 high mannose type N-glycan biosynthesis 1157:1195 high mannose type N-glycan biosynthesis 1157:1195 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 4 21 theme GSCX 713:716 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 7 22 theme xenograft 1313:1321 arg1 tissue 1323:1328 xenograft tissue 1313:1328 xenograft tissue 1313:1328 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 4 23 theme preclinical 673:683 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 0 24 theme Cell 64:67 arg1 Xenografts 69:78 Glioma Stem Cell Xenografts 52:78 Glioma Stem Cell Xenografts 52:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 9 25 theme molecular 1788:1796 arg1 determinates 1798:1809 molecular determinates 1788:1809 molecular determinates in BM-hMSC homing to glioma 1788:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 4 26 theme stem 692:695 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 6 27 from enrichment 1143:1152 arg1 phenotype 1218:1226 the non-attractor phenotype 1200:1226 the non-attractor phenotype 1200:1226 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 9 28 dep attractor 1730:1738 arg1 GSCXs 1758:1762 GSCXs 1758:1762 GSCXs 1758:1762 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 7 29 gly deglycosylation 1273:1287 arg1 validation 1240:1249 Orthogonal validation 1229:1249 Orthogonal validation 1229:1249 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 5 30 from tumor 1018:1022 arg1 engraftment 987:997 the engraftment 983:997 the engraftment of BM-hMSCs in the tumor 983:1022 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 4 31 theme unequal 754:760 arg1 tropism 762:768 unequal tropism 754:768 unequal tropism toward these tumors 754:788 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 0 32 theme Xenografts 69:78 arg1 Profiling 39:47 Transcriptomic and Glycomic Profiling 11:47 Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts 11:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 1 33 theme human 101:105 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 33 theme human 101:105 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 7 34 theme tumor 1296:1300 arg1 regions 1302:1308 the tumor regions 1292:1308 the tumor regions of xenograft tissue 1292:1328 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 35 theme sialic 1529:1534 arg1 N-glycans 1552:1560 terminal sialic acid-containing N-glycans 1520:1560 terminal sialic acid-containing N-glycans in non-attractors and terminal galactose 1520:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 4 36 theme GBM 728:730 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 9 37 from homing 1822:1827 arg1 scope 1779:1783 the scope 1775:1783 the scope of molecular determinates in BM-hMSC homing to glioma 1775:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 1 38 theme stem 119:122 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 38 theme stem 119:122 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 8 39 theme N-glycans 1552:1560 arg1 prevalence 1506:1515 the prevalence 1502:1515 the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors 1502:1650 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 2 40 theme GBM 375:377 arg1 treatment 379:387 GBM treatment 375:387 GBM treatment 375:387 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 26185906 7 41 theme F 1271:1271 arg1 deglycosylation 1273:1287 topical PNGase F deglycosylation 1256:1287 topical PNGase F deglycosylation 1256:1287 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 0 42 theme Glioma 52:57 arg1 Xenografts 69:78 Glioma Stem Cell Xenografts 52:78 Glioma Stem Cell Xenografts 52:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 7 43 theme topical 1256:1262 arg1 deglycosylation 1273:1287 topical PNGase F deglycosylation 1256:1287 topical PNGase F deglycosylation 1256:1287 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 44 theme terminal 1584:1591 arg1 galactose 1593:1601 terminal galactose 1584:1601 terminal galactose 1584:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 6 45 theme attractor 1115:1123 arg1 phenotype 1125:1133 the attractor phenotype 1111:1133 the attractor phenotype 1111:1133 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 9 46 from scope 1779:1783 arg1 homing 1822:1827 BM-hMSC homing 1814:1827 BM-hMSC homing to glioma 1814:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 0 47 theme Stem 59:62 arg1 Xenografts 69:78 Glioma Stem Cell Xenografts 52:78 Glioma Stem Cell Xenografts 52:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 8 48 theme N-glycans 1628:1636 arg1 prevalence 1506:1515 the prevalence 1502:1515 the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors 1502:1650 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 1 49 link marrow-derived 86:99 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 49 link marrow-derived 86:99 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 2 50 theme unique 257:262 arg1 property 264:271 this unique property 252:271 this unique property of BM-hMSCs 252:283 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 26185906 7 51 theme N-glycans 1409:1417 arg1 presence 1369:1376 the presence 1365:1376 the presence of increased high mannose type N-glycans in the non-attractors 1365:1439 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 52 from galactose 1593:1601 arg1 prevalence 1506:1515 the prevalence 1502:1515 the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors 1502:1650 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 3 53 theme complex 491:497 arg1 Extravasation 390:402 Extravasation 390:402 Extravasation 390:402 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 3 53 theme complex 491:497 arg1 process 499:505 a highly complex process 482:505 a highly complex process 482:505 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 9 54 theme differential 1696:1707 arg1 profiles 1718:1725 differential glycomic profiles 1696:1725 differential glycomic profiles in attractor and non-attractor GSCXs 1696:1762 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 7 55 theme mannose 1396:1402 arg1 N-glycans 1409:1417 increased high mannose type N-glycans 1381:1417 increased high mannose type N-glycans 1381:1417 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 1 56 contain have 141:144 arg2 ability 157:163 the innate ability 146:163 the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM) 146:238 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 56 contain have 141:144 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 56 contain have 141:144 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 6 57 theme non-attractor 1204:1216 arg1 phenotype 1218:1226 the non-attractor phenotype 1200:1226 the non-attractor phenotype 1200:1226 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 5 58 theme glycan 844:849 arg1 compositions 851:862 the glycan compositions 840:862 the glycan compositions 840:862 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 7 59 theme increased 1381:1389 arg1 N-glycans 1409:1417 increased high mannose type N-glycans 1381:1417 increased high mannose type N-glycans 1381:1417 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 6 60 theme significant 1078:1088 arg1 heterogeneity 1090:1102 significant heterogeneity 1078:1102 significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype 1078:1226 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 6 61 theme biosynthesis 1184:1195 arg1 phenotype 1125:1133 the attractor phenotype 1111:1133 the attractor phenotype 1111:1133 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 6 61 theme biosynthesis 1184:1195 arg1 enrichment 1143:1152 the enrichment 1139:1152 the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype 1139:1226 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 5 62 from differences 825:835 arg1 compositions 851:862 the glycan compositions 840:862 the glycan compositions 840:862 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 6 63 theme type 1170:1173 arg1 biosynthesis 1184:1195 high mannose type N-glycan biosynthesis 1157:1195 high mannose type N-glycan biosynthesis 1157:1195 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 4 64 theme cell 697:700 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 7 65 from presence 1369:1376 arg1 non-attractors 1426:1439 the non-attractors 1422:1439 the non-attractors 1422:1439 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 6 66 theme high 1157:1160 arg1 biosynthesis 1184:1195 high mannose type N-glycan biosynthesis 1157:1195 high mannose type N-glycan biosynthesis 1157:1195 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 7 67 theme Orthogonal 1229:1238 arg1 validation 1240:1249 Orthogonal validation 1229:1249 Orthogonal validation 1229:1249 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 5 68 from engraftment 987:997 arg1 tumor 1018:1022 the tumor 1014:1022 the tumor 1014:1022 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 8 69 theme glycomic 1478:1485 arg1 study 1487:1491 our glycomic study 1474:1491 our glycomic study 1474:1491 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 7 70 theme tissue 1323:1328 arg1 regions 1302:1308 the tumor regions 1292:1308 the tumor regions of xenograft tissue 1292:1328 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 4 71 theme significant 661:671 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 1 72 theme innate 150:155 arg1 ability 157:163 the innate ability 146:163 the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM) 146:238 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 9 73 theme determinates 1798:1809 arg1 scope 1779:1783 the scope 1775:1783 the scope of molecular determinates in BM-hMSC homing to glioma 1775:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 4 74 theme glioma 685:690 arg1 model 719:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model 643:723 a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM 643:730 However, in a translationally significant preclinical glioma stem cell xenograft (GSCX) model of GBM, BM-hMSCs demonstrate unequal tropism toward these tumors. 26185906 9 75 theme BM-hMSC 1814:1820 arg1 homing 1822:1827 BM-hMSC homing 1814:1827 BM-hMSC homing to glioma 1814:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 6 76 from phenotype 1125:1133 arg1 phenotype 1218:1226 the non-attractor phenotype 1200:1226 the non-attractor phenotype 1200:1226 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 5 77 from BM-hMSCs 1002:1009 arg1 tumor 1018:1022 the tumor 1014:1022 the tumor 1014:1022 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 8 78 theme terminal 1520:1527 arg1 N-glycans 1552:1560 terminal sialic acid-containing N-glycans 1520:1560 terminal sialic acid-containing N-glycans in non-attractors and terminal galactose 1520:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 2 79 dep cell-mediated 317:329 arg1 therapeutic 331:341 therapeutic 331:341 therapeutic 331:341 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 26185906 0 80 theme Transcriptomic 11:24 arg1 Profiling 39:47 Transcriptomic and Glycomic Profiling 11:47 Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts 11:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 5 81 theme attractors 903:912 arg1 homing 894:899 homing 894:899 homing ("attractors") 894:914 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 5 81 theme attractors 903:912 arg1 " 913:913 "attractors" 902:913 "attractors" 902:913 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 8 82 theme acid-containing 1536:1550 arg1 N-glycans 1552:1560 terminal sialic acid-containing N-glycans 1520:1560 terminal sialic acid-containing N-glycans in non-attractors and terminal galactose 1520:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 0 83 theme Glycomic 30:37 arg1 Profiling 39:47 Transcriptomic and Glycomic Profiling 11:47 Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts 11:78 Integrated Transcriptomic and Glycomic Profiling of Glioma Stem Cell Xenografts. 26185906 1 84 theme mesenchymal 107:117 arg1 BM-hMSCs 131:138 BM-hMSCs 131:138 BM-hMSCs 131:138 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 1 84 theme mesenchymal 107:117 arg1 cells 124:128 Bone marrow-derived human mesenchymal stem cells 81:128 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) 81:139 Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) have the innate ability to migrate or home toward and engraft in tumors such as glioblastoma (GBM). 26185906 2 85 theme treatment 379:387 arg1 advancement 360:370 the advancement 356:370 the advancement of GBM treatment 356:387 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 26185906 7 86 theme PNGase 1264:1269 arg1 deglycosylation 1273:1287 topical PNGase F deglycosylation 1256:1287 topical PNGase F deglycosylation 1256:1287 Orthogonal validation with topical PNGase F deglycosylation on the tumor regions of xenograft tissue, followed by nLC-ESI-MS, confirmed the presence of increased high mannose type N-glycans in the non-attractors. 26185906 8 87 from prevalence 1506:1515 arg1 attractors 1641:1650 attractors 1641:1650 attractors 1641:1650 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 8 87 from prevalence 1506:1515 arg1 non-attractors 1565:1578 non-attractors 1565:1578 non-attractors 1565:1578 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 8 87 from prevalence 1506:1515 arg1 galactose 1593:1601 terminal galactose 1584:1601 terminal galactose 1584:1601 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 9 88 from determinates 1798:1809 arg1 homing 1822:1827 BM-hMSC homing 1814:1827 BM-hMSC homing to glioma 1814:1837 Our results provide the first evidence for differential glycomic profiles in attractor and non-attractor GSCXs and extend the scope of molecular determinates in BM-hMSC homing to glioma. 26185906 6 89 theme glycotranscriptomic 1040:1058 arg1 analysis 1060:1067 glycotranscriptomic analysis 1040:1067 glycotranscriptomic analysis 1040:1067 In this study, glycotranscriptomic analysis revealed significant heterogeneity within the attractor phenotype and the enrichment of high mannose type N-glycan biosynthesis in the non-attractor phenotype. 26185906 8 90 theme N-acetyl-glucosamine 1607:1626 arg1 N-glycans 1628:1636 N-acetyl-glucosamine N-glycans 1607:1636 N-acetyl-glucosamine N-glycans 1607:1636 Additional evidence provided by our glycomic study revealed the prevalence of terminal sialic acid-containing N-glycans in non-attractors and terminal galactose and N-acetyl-glucosamine N-glycans in attractors. 26185906 3 91 theme BM-hMSCs—enter 452:465 arg1 tissue 471:476 BM-hMSCs—enter the tissue 452:476 BM-hMSCs—enter the tissue 452:476 Extravasation, the process by which blood-borne cells—such as BM-hMSCs—enter the tissue, is a highly complex process but is heavily dependent upon glycosylation for glycan-glycan and glycan-protein adhesion between the cell and endothelium. 26185906 5 92 theme non-attractors 940:953 arg1 GSCXs 876:880 the GSCXs 872:880 the GSCXs that elicit homing ("attractors") 872:914 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 5 92 theme non-attractors 940:953 arg1 " 954:954 "non-attractors" 939:954 "non-attractors" 939:954 We hypothesized that there may be differences in the glycan compositions between the GSCXs that elicit homing ("attractors") and those that do not ("non-attractors") that facilitate or impede the engraftment of BM-hMSCs in the tumor. 26185906 2 93 theme cell-mediated 317:329 arg1 delivery 343:350 cell-mediated therapeutic delivery 317:350 cell-mediated therapeutic delivery 317:350 Because of this unique property of BM-hMSCs, we have explored their use for cell-mediated therapeutic delivery for the advancement of GBM treatment. 28259967 11 0 theme MAL-I 1558:1562 arg1 staining 1546:1553 the staining 1542:1553 the staining of MAL-I in GC tissues 1542:1576 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 5 1 theme lectins 634:640 arg1 specificities 617:629 glycan-binding specificities 602:629 glycan-binding specificities of lectins 602:640 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 13 2 theme therapeutic 1867:1877 arg1 approaches 1879:1888 new therapeutic approaches 1863:1888 new therapeutic approaches for GC 1863:1895 This property may be important for developing new therapeutic approaches for GC. 28259967 9 3 theme decreased 1339:1347 arg1 level 1349:1353 a decreased level 1337:1353 a decreased level of α2-3Sia 1337:1364 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 6 4 theme metastatic 850:859 arg1 capacity 861:868 their metastatic capacity 844:868 their metastatic capacity 844:868 Importantly, we found that the binding of Maackia amurensis lectin-I (MAL-I) to GC cells was proportional to their metastatic capacity. 28259967 12 5 dep progression 1783:1793 arg1 the 1779:1781 the 1779:1781 the 1779:1781 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 11 6 theme high 1590:1593 arg1 expression 1595:1604 high expression 1590:1604 high expression of α2-3Sia 1590:1615 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 2 7 theme Aberrant 235:242 arg1 glycosylation 244:256 Aberrant glycosylation 235:256 Aberrant glycosylation 235:256 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 1 8 theme metastatic 156:165 arg1 disease 167:173 a highly metastatic disease 147:173 a highly metastatic disease 147:173 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 1 8 theme metastatic 156:165 arg1 cancer 132:137 Gastric cancer 124:137 Gastric cancer (GC) 124:142 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 4 9 theme human 537:541 arg1 SGC-7901 558:565 SGC-7901 558:565 SGC-7901 558:565 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 9 theme human 537:541 arg1 lines 551:555 the three human GC cell lines 527:555 the three human GC cell lines 527:555 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 9 theme human 537:541 arg1 MGC-803 580:586 MGC-803 580:586 MGC-803 580:586 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 9 theme human 537:541 arg1 BGC-823 568:574 BGC-823 568:574 BGC-823 568:574 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 2 10 theme molecular 273:281 arg1 changes 283:289 many molecular changes 268:289 many molecular changes that accompany malignant transformation 268:329 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 8 11 theme expression 1122:1131 arg1 levels 1133:1138 the mRNA and protein expression levels 1101:1138 the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, 1101:1241 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 12 12 from progression 1783:1793 arg1 understanding 1735:1747 better understanding 1728:1747 better understanding of the function of α2-3Sia in the progression and metastasis of GC 1728:1814 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 5 13 theme sugar 678:682 arg1 mannose 704:710 mannose 704:710 mannose 704:710 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 13 theme sugar 678:682 arg1 fucose 727:732 fucose 727:732 fucose 727:732 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 13 theme sugar 678:682 arg1 structures 684:693 common sugar structures 671:693 common sugar structures 671:693 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 13 theme sugar 678:682 arg1 galactose 713:721 galactose 713:721 galactose 713:721 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 9 14 theme invasion 1391:1398 arg1 ability 1380:1386 ability 1380:1386 ability of invasion and migration 1380:1412 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 1 15 theme leading 190:196 arg1 causes 198:203 the leading causes 186:203 the leading causes of cancer death in the world 186:232 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 8 16 theme α2,3-sialyltransferase IV  1157:1182 arg1 levels 1133:1138 the mRNA and protein expression levels 1101:1138 the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, 1101:1241 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 5 17 theme cell 650:653 arg1 lines 655:659 all GC cell lines 643:659 all GC cell lines 643:659 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 1 18 theme Gastric 124:130 arg1 cancer 132:137 Gastric cancer 124:137 Gastric cancer (GC) 124:142 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 1 18 theme Gastric 124:130 arg1 disease 167:173 a highly metastatic disease 147:173 a highly metastatic disease 147:173 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 1 18 theme Gastric 124:130 arg1 GC 140:141 GC 140:141 GC 140:141 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 8 19 theme α2-3Sia 1234:1240 arg1 synthesis 1221:1229 the synthesis 1217:1229 the synthesis of α2-3Sia 1217:1240 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 11 20 theme TNM 1668:1670 arg1 stage 1672:1676 TNM stage 1668:1676 TNM stage 1668:1676 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 0 21 theme cancer 65:70 arg1 metastasis 77:86 gastric cancer cell metastasis 57:86 gastric cancer cell metastasis 57:86 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 7 22 theme acids  934:939 arg1 level 906:910 the level 902:910 the level 902:910 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 10 23 from expression 1425:1434 arg1 cells 1461:1465 SGC-7901 cells 1452:1465 SGC-7901 cells 1452:1465 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 11 24 theme overall 1687:1693 arg1 survival 1695:1702 poor overall survival 1682:1702 poor overall survival 1682:1702 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 3 25 from changes 391:397 arg1 mechanisms 435:444 its potential mechanisms 421:444 its potential mechanisms 421:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 3 25 from changes 391:397 arg1 progression 405:415 GC progression 402:415 GC progression 402:415 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 7 26 theme α2,3-linked 915:925 arg1 α2-3Sia 941:947 α2-3Sia 941:947 α2-3Sia 941:947 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 7 26 theme α2,3-linked 915:925 arg1 acids  934:939 α2,3-linked sialic acids  915:939 α2,3-linked sialic acids (α2-3Sia) 915:948 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 8 27 theme MGC-803 1275:1281 arg1 cells 1283:1287 MGC-803 cells 1275:1287 MGC-803 cells 1275:1287 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 7 28 link α2,3-linked 915:925 arg1 α2-3Sia 941:947 α2-3Sia 941:947 α2-3Sia 941:947 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 7 28 link α2,3-linked 915:925 arg1 acids  934:939 α2,3-linked sialic acids  915:939 α2,3-linked sialic acids (α2-3Sia) 915:948 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 12 29 from metastasis 1799:1808 arg1 understanding 1735:1747 better understanding 1728:1747 better understanding of the function of α2-3Sia in the progression and metastasis of GC 1728:1814 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 0 30 theme poor 108:111 arg1 prognosis 113:121 poor prognosis 108:121 poor prognosis 108:121 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 3 31 from identification 356:369 arg1 mechanisms 435:444 its potential mechanisms 421:444 its potential mechanisms 421:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 3 31 from identification 356:369 arg1 progression 405:415 GC progression 402:415 GC progression 402:415 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 3 32 theme GC 402:403 arg1 progression 405:415 GC progression 402:415 GC progression 402:415 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 0 33 theme Enhanced 0:7 arg1 expression 9:18 Enhanced expression 0:18 Enhanced expression of α2,3-linked sialic acids 0:46 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 12 34 theme GC 1813:1814 arg1 metastasis 1799:1808 metastasis 1799:1808 metastasis 1799:1808 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 12 34 theme GC 1813:1814 arg1 progression 1783:1793 progression 1783:1793 progression 1783:1793 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 3 35 theme changes 391:397 arg1 identification 356:369 identification 356:369 identification of glycan profiling changes in GC progression and its potential mechanisms 356:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 4 36 theme GC 543:544 arg1 SGC-7901 558:565 SGC-7901 558:565 SGC-7901 558:565 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 36 theme GC 543:544 arg1 lines 551:555 the three human GC cell lines 527:555 the three human GC cell lines 527:555 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 36 theme GC 543:544 arg1 MGC-803 580:586 MGC-803 580:586 MGC-803 580:586 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 36 theme GC 543:544 arg1 BGC-823 568:574 BGC-823 568:574 BGC-823 568:574 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 10 37 theme Exogenous 1415:1423 arg1 expression 1425:1434 Exogenous expression 1415:1434 Exogenous expression of ST3Gal-IV in SGC-7901 cells 1415:1465 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 3 38 theme potential 425:433 arg1 mechanisms 435:444 its potential mechanisms 421:444 its potential mechanisms 421:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 0 39 theme sialic 35:40 arg1 acids 42:46 α2,3-linked sialic acids 23:46 α2,3-linked sialic acids 23:46 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 12 40 theme better 1728:1733 arg1 understanding 1735:1747 better understanding 1728:1747 better understanding of the function of α2-3Sia in the progression and metastasis of GC 1728:1814 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 3 41 theme glycan 374:379 arg1 changes 391:397 glycan profiling changes 374:397 glycan profiling changes in GC progression and its potential mechanisms 374:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 4 42 theme differential 503:514 arg1 glycans 516:522 the differential glycans 499:522 the differential glycans 499:522 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 11 43 theme α2-3Sia 1609:1615 arg1 expression 1595:1604 high expression 1590:1604 high expression of α2-3Sia 1590:1615 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 3 44 from mechanisms 435:444 arg1 identification 356:369 identification 356:369 identification of glycan profiling changes in GC progression and its potential mechanisms 356:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 7 45 theme SGC-7901 1072:1079 arg1 cells 1081:1085 SGC-7901 cells 1072:1085 SGC-7901 cells 1072:1085 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 9 46 theme ST3Gal-IV 1303:1311 arg1 Knockdown 1290:1298 Knockdown 1290:1298 Knockdown of ST3Gal-IV in MGC-803 cells 1290:1328 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 12 47 from understanding 1735:1747 arg1 metastasis 1799:1808 metastasis 1799:1808 metastasis 1799:1808 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 12 47 from understanding 1735:1747 arg1 progression 1783:1793 progression 1783:1793 progression 1783:1793 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 11 48 theme node 1651:1654 arg1 metastasis 1656:1665 lymph node metastasis 1645:1665 lymph node metastasis 1645:1665 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 7 49 theme Further 871:877 arg1 analysis 879:886 Further analysis 871:886 Further analysis 871:886 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 9 50 theme MGC-803 1316:1322 arg1 cells 1324:1328 MGC-803 cells 1316:1328 MGC-803 cells 1316:1328 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 6 51 theme amurensis 785:793 arg1 binding 766:772 the binding 762:772 the binding of Maackia amurensis lectin-I (MAL-I) to GC cells 762:822 Importantly, we found that the binding of Maackia amurensis lectin-I (MAL-I) to GC cells was proportional to their metastatic capacity. 28259967 6 51 theme amurensis 785:793 arg1 proportional 828:839 proportional 828:839 proportional 828:839 Importantly, we found that the binding of Maackia amurensis lectin-I (MAL-I) to GC cells was proportional to their metastatic capacity. 28259967 3 52 from progression 405:415 arg1 identification 356:369 identification 356:369 identification of glycan profiling changes in GC progression and its potential mechanisms 356:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 1 53 theme cancer 208:213 arg1 death 215:219 cancer death 208:219 cancer death 208:219 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 10 54 theme cell 1476:1479 arg1 migration 1481:1489 cell migration 1476:1489 cell migration 1476:1489 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 7 55 theme MGC-803 1020:1026 arg1 cells 1028:1032 MGC-803 cells 1020:1032 MGC-803 cells 1020:1032 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 0 56 link α2,3-linked 23:33 arg1 acids 42:46 α2,3-linked sialic acids 23:46 α2,3-linked sialic acids 23:46 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 7 57 theme low 1041:1043 arg1 expression 1045:1054 low expression 1041:1054 low expression 1041:1054 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 9 58 theme α2-3Sia 1358:1364 arg1 level 1349:1353 a decreased level 1337:1353 a decreased level of α2-3Sia 1337:1364 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 5 59 theme glycan-binding 602:615 arg1 specificities 617:629 glycan-binding specificities 602:629 glycan-binding specificities of lectins 602:640 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 2 60 theme changes 283:289 arg1 one 261:263 one 261:263 one 261:263 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 2 60 theme changes 283:289 arg1 changes 283:289 many molecular changes 268:289 many molecular changes that accompany malignant transformation 268:329 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 11 61 from staining 1546:1553 arg1 tissues 1570:1576 GC tissues 1567:1576 GC tissues 1567:1576 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 8 62 theme mRNA 1105:1108 arg1 levels 1133:1138 the mRNA and protein expression levels 1101:1138 the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, 1101:1241 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 10 63 theme α2-3Sia 1520:1526 arg1 migration 1481:1489 cell migration 1476:1489 cell migration 1476:1489 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 10 63 theme α2-3Sia 1520:1526 arg1 invasion 1492:1499 invasion 1492:1499 invasion 1492:1499 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 10 63 theme α2-3Sia 1520:1526 arg1 content 1509:1515 the content 1505:1515 the content of α2-3Sia 1505:1526 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 5 64 theme common 671:676 arg1 mannose 704:710 mannose 704:710 mannose 704:710 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 64 theme common 671:676 arg1 fucose 727:732 fucose 727:732 fucose 727:732 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 64 theme common 671:676 arg1 structures 684:693 common sugar structures 671:693 common sugar structures 671:693 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 5 64 theme common 671:676 arg1 galactose 713:721 galactose 713:721 galactose 713:721 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 13 65 theme new 1863:1865 arg1 approaches 1879:1888 new therapeutic approaches 1863:1888 new therapeutic approaches for GC 1863:1895 This property may be important for developing new therapeutic approaches for GC. 28259967 2 66 theme many 268:271 arg1 changes 283:289 many molecular changes 268:289 many molecular changes that accompany malignant transformation 268:329 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 8 67 theme protein 1114:1120 arg1 levels 1133:1138 the mRNA and protein expression levels 1101:1138 the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, 1101:1241 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 5 68 theme GC 647:648 arg1 lines 655:659 all GC cell lines 643:659 all GC cell lines 643:659 According to glycan-binding specificities of lectins, all GC cell lines expressed common sugar structures, such as mannose, galactose and fucose. 28259967 8 69 theme β-galactoside 1143:1155 arg1 ST3Gal-IV 1184:1192 ST3Gal-IV 1184:1192 ST3Gal-IV 1184:1192 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 8 69 theme β-galactoside 1143:1155 arg1 α2,3-sialyltransferase IV  1157:1182 β-galactoside α2,3-sialyltransferase IV  1143:1182 β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV) 1143:1193 In addition, the mRNA and protein expression levels of β-galactoside α2,3-sialyltransferase IV (ST3Gal-IV), which was related to the synthesis of α2-3Sia, were substantially increased in MGC-803 cells. 28259967 12 70 from function 1756:1763 arg1 metastasis 1799:1808 metastasis 1799:1808 metastasis 1799:1808 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 12 70 from function 1756:1763 arg1 progression 1783:1793 progression 1783:1793 progression 1783:1793 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 1 71 from causes 198:203 arg1 world 228:232 the world 224:232 the world 224:232 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 0 72 theme cell 72:75 arg1 metastasis 77:86 gastric cancer cell metastasis 57:86 gastric cancer cell metastasis 57:86 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 1 73 theme causes 198:203 arg1 causes 198:203 the leading causes 186:203 the leading causes of cancer death in the world 186:232 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 1 73 theme causes 198:203 arg1 one 179:181 one 179:181 one 179:181 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28259967 7 74 located detected 1060:1067 arg1 cells 1081:1085 SGC-7901 cells 1072:1085 SGC-7901 cells 1072:1085 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 7 74 located detected 1060:1067 arg2 expression 1045:1054 low expression 1041:1054 low expression 1041:1054 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 9 75 from Knockdown 1290:1298 arg1 cells 1324:1328 MGC-803 cells 1316:1328 MGC-803 cells 1316:1328 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 7 76 theme sialic 927:932 arg1 α2-3Sia 941:947 α2-3Sia 941:947 α2-3Sia 941:947 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 7 76 theme sialic 927:932 arg1 acids  934:939 α2,3-linked sialic acids  915:939 α2,3-linked sialic acids (α2-3Sia) 915:948 Further analysis revealed that the level of α2,3-linked sialic acids (α2-3Sia), which can be recognized by MAL-I, was significantly overexpressed in MGC-803 cells, while low expression was detected in SGC-7901 cells. 28259967 11 77 theme poor 1682:1685 arg1 survival 1695:1702 poor overall survival 1682:1702 poor overall survival 1682:1702 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 6 78 dep amurensis 785:793 arg1 lectin-I  795:803 lectin-I  795:803 lectin-I  795:803 Importantly, we found that the binding of Maackia amurensis lectin-I (MAL-I) to GC cells was proportional to their metastatic capacity. 28259967 0 79 theme α2,3-linked 23:33 arg1 acids 42:46 α2,3-linked sialic acids 23:46 α2,3-linked sialic acids 23:46 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 4 80 theme cell 546:549 arg1 SGC-7901 558:565 SGC-7901 558:565 SGC-7901 558:565 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 80 theme cell 546:549 arg1 lines 551:555 the three human GC cell lines 527:555 the three human GC cell lines 527:555 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 80 theme cell 546:549 arg1 MGC-803 580:586 MGC-803 580:586 MGC-803 580:586 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 4 80 theme cell 546:549 arg1 BGC-823 568:574 BGC-823 568:574 BGC-823 568:574 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 0 81 theme acids 42:46 arg1 expression 9:18 Enhanced expression 0:18 Enhanced expression of α2,3-linked sialic acids 0:46 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 0 82 theme gastric 57:63 arg1 metastasis 77:86 gastric cancer cell metastasis 57:86 gastric cancer cell metastasis 57:86 Enhanced expression of α2,3-linked sialic acids promotes gastric cancer cell metastasis and correlates with poor prognosis. 28259967 12 83 theme function 1756:1763 arg1 understanding 1735:1747 better understanding 1728:1747 better understanding of the function of α2-3Sia in the progression and metastasis of GC 1728:1814 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 3 84 theme profiling 381:389 arg1 changes 391:397 glycan profiling changes 374:397 glycan profiling changes in GC progression and its potential mechanisms 374:444 This study was aimed at identification of glycan profiling changes in GC progression and its potential mechanisms. 28259967 10 85 theme SGC-7901 1452:1459 arg1 cells 1461:1465 SGC-7901 cells 1452:1465 SGC-7901 cells 1452:1465 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 11 86 theme lymph 1645:1649 arg1 node 1651:1654 lymph node 1645:1654 lymph node metastasis 1645:1665 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 2 87 theme malignant 306:314 arg1 transformation 316:329 malignant transformation 306:329 malignant transformation 306:329 Aberrant glycosylation is one of many molecular changes that accompany malignant transformation. 28259967 12 88 theme α2-3Sia 1768:1774 arg1 function 1756:1763 the function 1752:1763 the function of α2-3Sia in the progression and metastasis of GC 1752:1814 These findings lead to better understanding of the function of α2-3Sia in the progression and metastasis of GC. 28259967 10 89 theme ST3Gal-IV 1439:1447 arg1 expression 1425:1434 Exogenous expression 1415:1434 Exogenous expression of ST3Gal-IV in SGC-7901 cells 1415:1465 Exogenous expression of ST3Gal-IV in SGC-7901 cells enhanced cell migration, invasion and the content of α2-3Sia. 28259967 9 90 theme migration 1404:1412 arg1 ability 1380:1386 ability 1380:1386 ability of invasion and migration 1380:1412 Knockdown of ST3Gal-IV in MGC-803 cells led to a decreased level of α2-3Sia and decreased ability of invasion and migration. 28259967 11 91 theme GC 1567:1568 arg1 tissues 1570:1576 GC tissues 1567:1576 GC tissues 1567:1576 Furthermore, the staining of MAL-I in GC tissues showed that high expression of α2-3Sia was closely correlated with lymph node metastasis, TNM stage and poor overall survival. 28259967 4 92 with microarray 461:470 arg1 91 lectins 477:486 91 lectins 477:486 91 lectins 477:486 We employed a microarray with 91 lectins to compare the differential glycans in the three human GC cell lines, SGC-7901, BGC-823 and MGC-803. 28259967 1 93 theme death 215:219 arg1 causes 198:203 the leading causes 186:203 the leading causes of cancer death in the world 186:232 Gastric cancer (GC) is a highly metastatic disease and one of the leading causes of cancer death in the world. 28738643 5 0 theme surface 791:797 arg1 544.99m2g-1 805:815 544.99m2g-1 805:815 544.99m2g-1 805:815 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 0 theme surface 791:797 arg1 area 799:802 high BET surface area 782:802 high BET surface area (544.99m2g-1) 782:816 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 11 1 theme adsorbent 1637:1645 arg1 material 1647:1654 a promising adsorbent material 1625:1654 a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis 1625:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 11 2 link N-linked 1556:1563 arg1 glycans 1565:1571 N-linked glycans 1556:1571 N-linked glycans 1556:1571 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 10 3 link N-linked 1439:1446 arg1 glycans 1448:1454 31 N-linked glycans 1436:1454 31 N-linked glycans in total 1436:1463 Additionally, 31 N-linked glycans in total were enriched from human serum. 28738643 1 4 theme high 272:275 arg1 contents 296:303 high graphitized carbon contents 272:303 high graphitized carbon contents 272:303 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 5 theme composites 87:96 arg1 preparation 7:17 Facile preparation 0:17 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 0:172 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 7 6 theme human 1096:1100 arg1 serum 1102:1106 human serum 1096:1106 human serum 1096:1106 By taking advantage of the interactions between carbon and glycans, FLOMC-GO was utilized to enrich N-linked glycans from OVA and human serum. 28738643 1 7 theme Novel 174:178 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 7 theme Novel 174:178 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 8 theme graphitized 277:287 arg1 contents 296:303 high graphitized carbon contents 272:303 high graphitized carbon contents 272:303 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 9 theme application 106:116 arg1 preparation 7:17 Facile preparation 0:17 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 0:172 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 5 10 theme prepared 708:715 arg1 FLOMC-GO 717:724 The prepared FLOMC-GO 704:724 The prepared FLOMC-GO 704:724 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 1 11 theme carbon 289:294 arg1 contents 296:303 high graphitized carbon contents 272:303 high graphitized carbon contents 272:303 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 5 12 theme large 750:754 arg1 1.03cm3g-1 769:778 1.03cm3g-1 769:778 1.03cm3g-1 769:778 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 12 theme large 750:754 arg1 volume 761:766 a large pore volume 748:766 a large pore volume (1.03cm3g-1) 748:779 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 11 13 theme easy 1512:1515 arg1 synthesis 1517:1525 The relatively easy synthesis 1497:1525 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency 1497:1608 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 0 14 theme selective 122:130 arg1 enrichment 132:141 selective enrichment 122:141 selective enrichment of N-glycans from human serum 122:171 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 3 15 theme 3-D 518:520 arg1 structure 522:530 a 3-D structure 516:530 a 3-D structure 516:530 Then, the sandwich-like composites were further combined together to form a 3-D structure through pre-carbonization and carbonization. 28738643 5 16 theme narrow 863:868 arg1 pore-size 870:878 a narrow pore-size 861:878 a narrow pore-size concentrated at 3.74nm 861:901 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 1 17 theme 3-D 180:182 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 17 theme 3-D 180:182 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 8 18 theme N-glycans 1182:1190 arg1 enrichment 1168:1177 the enrichment 1164:1177 the enrichment of N-glycans released from OVA 1164:1208 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 8 19 theme N-linked 1218:1225 arg1 glycans 1227:1233 25 N-linked glycans 1215:1233 25 N-linked glycans 1215:1233 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 5 20 theme mesoporous 835:844 arg1 structure 846:854 well-ordered mesoporous structure 822:854 well-ordered mesoporous structure 822:854 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 1 21 theme floor-like 184:193 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 21 theme floor-like 184:193 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 5 22 theme well-ordered 822:833 arg1 structure 846:854 well-ordered mesoporous structure 822:854 well-ordered mesoporous structure 822:854 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 7 23 theme N-linked 1066:1073 arg1 glycans 1075:1081 N-linked glycans 1066:1081 N-linked glycans 1066:1081 By taking advantage of the interactions between carbon and glycans, FLOMC-GO was utilized to enrich N-linked glycans from OVA and human serum. 28738643 5 24 theme high 782:785 arg1 544.99m2g-1 805:815 544.99m2g-1 805:815 544.99m2g-1 805:815 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 24 theme high 782:785 arg1 area 799:802 high BET surface area 782:802 high BET surface area (544.99m2g-1) 782:816 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 11 25 dep ability 1538:1544 arg1 enrich 1549:1554 enrich 1549:1554 to enrich N-linked glycans with high selectivity and efficiency 1546:1608 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 6 26 theme carbon 919:924 arg1 content 908:914 The content 904:914 The content of carbon 904:924 The content of carbon reached 80% and was highly graphitized. 28738643 6 26 theme carbon 919:924 arg1 graphitized 953:963 graphitized 953:963 graphitized 953:963 The content of carbon reached 80% and was highly graphitized. 28738643 1 27 theme ordered 195:201 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 27 theme ordered 195:201 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 28 theme Facile 0:5 arg1 preparation 7:17 Facile preparation 0:17 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 0:172 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 9 29 theme active 1327:1332 arg1 carbon 1334:1339 active carbon 1327:1339 active carbon 1327:1339 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 10 30 theme N-linked 1439:1446 arg1 glycans 1448:1454 31 N-linked glycans 1436:1454 31 N-linked glycans in total 1436:1463 Additionally, 31 N-linked glycans in total were enriched from human serum. 28738643 8 31 link N-linked 1218:1225 arg1 glycans 1227:1233 25 N-linked glycans 1215:1233 25 N-linked glycans 1215:1233 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 2 32 theme soft 427:430 arg1 template 432:439 the silica soft template 416:439 the silica soft template 416:439 The one-pot sol-gel method was employed to prepare the silica soft template. 28738643 1 33 theme mesoporous 203:212 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 33 theme mesoporous 203:212 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 34 theme 3-D 22:24 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 9 35 theme carbon 1334:1339 arg1 result 1317:1322 the result 1313:1322 the result of active carbon 1313:1339 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 10 36 theme human 1484:1488 arg1 serum 1490:1494 human serum 1484:1494 human serum 1484:1494 Additionally, 31 N-linked glycans in total were enriched from human serum. 28738643 1 37 theme carbon 214:219 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 37 theme carbon 214:219 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 38 with composites 245:254 arg1 contents 296:303 high graphitized carbon contents 272:303 high graphitized carbon contents 272:303 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 11 39 theme serum 1683:1687 arg1 biomarkers 1689:1698 human serum biomarkers 1677:1698 human serum biomarkers for disease diagnosis 1677:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 8 40 attach released 1192:1199 arg2 N-glycans 1182:1190 N-glycans 1182:1190 N-glycans released from OVA 1182:1208 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 8 40 attach released 1192:1199 arg1 OVA 1206:1208 OVA 1206:1208 OVA 1206:1208 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 4 41 theme sulfide 620:626 arg1 bridges 628:634 the sulphonyl and sulfide bridges 602:634 the sulphonyl and sulfide bridges 602:634 During these procedures, the sulphonyl and sulfide bridges were formed by cross-linking processes to connect the phenyl rings. 28738643 1 42 theme functionalized 221:234 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 42 theme functionalized 221:234 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 43 theme ordered 37:43 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 4 44 theme sulphonyl 606:614 arg1 bridges 628:634 the sulphonyl and sulfide bridges 602:634 the sulphonyl and sulfide bridges 602:634 During these procedures, the sulphonyl and sulfide bridges were formed by cross-linking processes to connect the phenyl rings. 28738643 5 45 theme pore 756:759 arg1 1.03cm3g-1 769:778 1.03cm3g-1 769:778 1.03cm3g-1 769:778 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 45 theme pore 756:759 arg1 volume 761:766 a large pore volume 748:766 a large pore volume (1.03cm3g-1) 748:779 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 11 46 theme biomarkers 1689:1698 arg1 discovery 1664:1672 the discovery 1660:1672 the discovery of human serum biomarkers for disease diagnosis 1660:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 3 47 theme sandwich-like 452:464 arg1 composites 466:475 the sandwich-like composites 448:475 the sandwich-like composites 448:475 Then, the sandwich-like composites were further combined together to form a 3-D structure through pre-carbonization and carbonization. 28738643 1 48 theme graphene 236:243 arg1 FLOMC-GO 257:264 FLOMC-GO 257:264 FLOMC-GO 257:264 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 1 48 theme graphene 236:243 arg1 composites 245:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites 174:254 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 49 theme floor-like 26:35 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 5 50 contain have 743:746 arg2 structure 846:854 well-ordered mesoporous structure 822:854 well-ordered mesoporous structure 822:854 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 50 contain have 743:746 arg2 1.03cm3g-1 769:778 1.03cm3g-1 769:778 1.03cm3g-1 769:778 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 50 contain have 743:746 arg2 544.99m2g-1 805:815 544.99m2g-1 805:815 544.99m2g-1 805:815 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 50 contain have 743:746 arg2 volume 761:766 a large pore volume 748:766 a large pore volume (1.03cm3g-1) 748:779 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 50 contain have 743:746 arg2 area 799:802 high BET surface area 782:802 high BET surface area (544.99m2g-1) 782:816 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 50 contain have 743:746 arg1 FLOMC-GO 717:724 The prepared FLOMC-GO 704:724 The prepared FLOMC-GO 704:724 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 0 51 theme N-glycans 146:154 arg1 enrichment 132:141 selective enrichment 122:141 selective enrichment of N-glycans from human serum 122:171 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 4 52 theme phenyl 690:695 arg1 rings 697:701 the phenyl rings 686:701 the phenyl rings 686:701 During these procedures, the sulphonyl and sulfide bridges were formed by cross-linking processes to connect the phenyl rings. 28738643 0 53 theme carbon 56:61 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 1 54 theme soft 343:346 arg1 method 357:362 a soft template method 341:362 a soft template method 341:362 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 5 55 theme BET 787:789 arg1 544.99m2g-1 805:815 544.99m2g-1 805:815 544.99m2g-1 805:815 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 5 55 theme BET 787:789 arg1 area 799:802 high BET surface area 782:802 high BET surface area (544.99m2g-1) 782:816 The prepared FLOMC-GO was confirmed to have a large pore volume (1.03cm3g-1), high BET surface area (544.99m2g-1) and well-ordered mesoporous structure with a narrow pore-size concentrated at 3.74nm. 28738643 11 56 theme high 1578:1581 arg1 selectivity 1583:1593 high selectivity 1578:1593 high selectivity 1578:1593 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 2 57 theme sol-gel 377:383 arg1 method 385:390 The one-pot sol-gel method 365:390 The one-pot sol-gel method 365:390 The one-pot sol-gel method was employed to prepare the silica soft template. 28738643 0 58 theme mesoporous 45:54 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 1 59 theme template 348:355 arg1 method 357:362 a soft template method 341:362 a soft template method 341:362 Novel 3-D floor-like ordered mesoporous carbon functionalized graphene composites, FLOMC-GO, with high graphitized carbon contents were successfully synthesized using a soft template method. 28738643 0 60 theme human 161:165 arg1 serum 167:171 human serum 161:171 human serum 161:171 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 11 61 theme N-linked 1556:1563 arg1 glycans 1565:1571 N-linked glycans 1556:1571 N-linked glycans 1556:1571 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 2 62 theme one-pot 369:375 arg1 method 385:390 The one-pot sol-gel method 365:390 The one-pot sol-gel method 365:390 The one-pot sol-gel method was employed to prepare the silica soft template. 28738643 9 63 theme OVA 1361:1363 arg1 digestion 1365:1373 OVA digestion 1361:1373 OVA digestion 1361:1373 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 9 64 theme glycans 1269:1275 arg1 times 1304:1308 7 times 1302:1308 7 times 1302:1308 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 9 64 theme glycans 1269:1275 arg1 intensity 1256:1264 The intensity 1252:1264 The intensity of glycans enriched by FLOMC-GO 1252:1296 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 7 65 used utilized 1047:1054 arg2 FLOMC-GO 1034:1041 FLOMC-GO 1034:1041 FLOMC-GO 1034:1041 By taking advantage of the interactions between carbon and glycans, FLOMC-GO was utilized to enrich N-linked glycans from OVA and human serum. 28738643 4 66 theme cross-linking 651:663 arg1 processes 665:673 cross-linking processes 651:673 cross-linking processes 651:673 During these procedures, the sulphonyl and sulfide bridges were formed by cross-linking processes to connect the phenyl rings. 28738643 8 67 theme excellent 1122:1130 arg1 size-exclusion 1132:1145 excellent size-exclusion 1122:1145 excellent size-exclusion 1122:1145 As expected, excellent size-exclusion was found during the enrichment of N-glycans released from OVA, and 25 N-linked glycans were identified. 28738643 0 68 from serum 167:171 arg1 enrichment 132:141 selective enrichment 122:141 selective enrichment of N-glycans from human serum 122:171 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 7 69 link N-linked 1066:1073 arg1 glycans 1075:1081 N-linked glycans 1066:1081 N-linked glycans 1066:1081 By taking advantage of the interactions between carbon and glycans, FLOMC-GO was utilized to enrich N-linked glycans from OVA and human serum. 28738643 0 70 theme graphene 78:85 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 11 71 theme disease 1704:1710 arg1 diagnosis 1712:1720 disease diagnosis 1704:1720 disease diagnosis 1704:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 10 72 from glycans 1448:1454 arg1 total 1459:1463 total 1459:1463 total 1459:1463 Additionally, 31 N-linked glycans in total were enriched from human serum. 28738643 2 73 theme silica 420:425 arg1 template 432:439 the silica soft template 416:439 the silica soft template 416:439 The one-pot sol-gel method was employed to prepare the silica soft template. 28738643 9 74 theme BSA 1378:1380 arg1 proteins 1394:1401 BSA interfering proteins 1378:1401 BSA interfering proteins 1378:1401 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 0 75 theme functionalized 63:76 arg1 composites 87:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 3-D floor-like ordered mesoporous carbon functionalized graphene composites 22:96 Facile preparation of 3-D floor-like ordered mesoporous carbon functionalized graphene composites and its application for selective enrichment of N-glycans from human serum. 28738643 11 76 theme promising 1627:1635 arg1 material 1647:1654 a promising adsorbent material 1625:1654 a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis 1625:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 7 77 theme interactions 993:1004 arg1 advantage 976:984 advantage 976:984 advantage of the interactions between carbon and glycans 976:1031 By taking advantage of the interactions between carbon and glycans, FLOMC-GO was utilized to enrich N-linked glycans from OVA and human serum. 28738643 9 78 theme digestion 1365:1373 arg1 ratio 1352:1356 the ratio 1348:1356 the ratio of OVA digestion to BSA interfering proteins 1348:1401 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 28738643 11 79 theme human 1677:1681 arg1 biomarkers 1689:1698 human serum biomarkers 1677:1698 human serum biomarkers for disease diagnosis 1677:1720 The relatively easy synthesis as well as ability to enrich N-linked glycans with high selectivity and efficiency makes FLOMC-GO a promising adsorbent material for the discovery of human serum biomarkers for disease diagnosis. 28738643 9 80 theme interfering 1382:1392 arg1 proteins 1394:1401 BSA interfering proteins 1378:1401 BSA interfering proteins 1378:1401 The intensity of glycans enriched by FLOMC-GO was 7 times to the result of active carbon, while the ratio of OVA digestion to BSA interfering proteins increased to 1:50. 27206480 2 0 theme flagellate 522:531 arg1 Enterobacteriaceae 533:550 the peritrichously flagellate Enterobacteriaceae 503:550 the peritrichously flagellate Enterobacteriaceae 503:550 However to date, despite extensive physiological and genetic characterization, it has never been reported for the peritrichously flagellate Enterobacteriaceae. 27206480 6 1 theme distinct 1312:1319 arg1 taxa 1321:1324 several distinct taxa 1304:1324 several distinct taxa within the family 1304:1342 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 4 2 theme transfer 936:943 arg1 evidence 908:915 evidence 908:915 evidence of horizontal gene transfer occurring within the FGI loci 908:973 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 2 3 theme peritrichously 507:520 arg1 Enterobacteriaceae 533:550 the peritrichously flagellate Enterobacteriaceae 503:550 the peritrichously flagellate Enterobacteriaceae 503:550 However to date, despite extensive physiological and genetic characterization, it has never been reported for the peritrichously flagellate Enterobacteriaceae. 27206480 8 4 theme glycosylation 1562:1574 arg1 loci 1592:1595 glycosylation and methylation loci 1562:1595 loci 1592:1595 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 4 5 theme gene 931:934 arg1 transfer 936:943 horizontal gene transfer 920:943 horizontal gene transfer occurring within the FGI loci 920:973 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 3 6 theme family 776:781 arg1 members 760:766 members 760:766 members of this family 760:781 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 3 7 theme comparative 567:577 arg1 approaches 587:596 comparative genomic approaches 567:596 comparative genomic approaches 567:596 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 4 8 theme FGIs 824:827 arg1 rest 837:840 the rest 833:840 the rest of the genome 833:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 8 theme FGIs 824:827 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 1 9 theme flagellar 285:293 arg1 biosynthesis 295:306 flagellar biosynthesis 285:306 flagellar biosynthesis 285:306 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 8 10 theme methylation 1580:1590 arg1 loci 1592:1595 glycosylation and methylation loci 1562:1595 loci 1592:1595 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 6 11 theme enterobacterial 1231:1245 arg1 pathogen 1247:1254 the enterobacterial pathogen 1227:1254 the enterobacterial pathogen Salmonella enterica 1227:1274 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 4 12 attach presence 864:871 arg2 elements 891:898 mobile genetic elements 876:898 mobile genetic elements 876:898 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 12 attach presence 864:871 arg1 rest 837:840 the rest 833:840 the rest of the genome 833:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 12 attach presence 864:871 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 2 13 theme extensive 418:426 arg1 characterization 454:469 extensive physiological and genetic characterization 418:469 extensive physiological and genetic characterization 418:469 However to date, despite extensive physiological and genetic characterization, it has never been reported for the peritrichously flagellate Enterobacteriaceae. 27206480 8 14 theme flagellin 1528:1536 arg1 loci 1551:1554 flagellin modification loci 1528:1554 flagellin modification loci 1528:1554 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 5 15 theme distinct 1054:1061 arg1 backbones 1069:1077 distinct sugar backbones 1054:1077 distinct sugar backbones 1054:1077 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 8 16 theme loci 1551:1554 arg1 prevalence 1498:1507 prevalence 1498:1507 prevalence 1498:1507 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 8 16 theme loci 1551:1554 arg1 versatility 1513:1523 versatility 1513:1523 versatility 1513:1523 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 4 17 theme horizontal 920:929 arg1 transfer 936:943 horizontal gene transfer 920:943 horizontal gene transfer occurring within the FGI loci 920:973 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 7 18 theme diverse 1464:1470 arg1 lineages 1472:1479 these diverse lineages 1458:1479 these diverse lineages 1458:1479 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 1 19 theme Gram-negative 208:220 arg1 bacteria 222:229 Gram-negative bacteria 208:229 Gram-negative bacteria 208:229 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 0 20 theme Flashy 0:5 arg1 flagella 7:14 Flashy flagella 0:14 Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae. 0:110 Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae. 27206480 4 21 theme G + C 803:807 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 8 22 theme modification 1538:1549 arg1 loci 1551:1554 flagellin modification loci 1528:1554 flagellin modification loci 1528:1554 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 3 23 theme Enterobacteriaceae 642:659 arg1 genomes 631:637 2,000 representative genomes 610:637 2,000 representative genomes of Enterobacteriaceae 610:659 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 7 24 theme FGI 1410:1412 arg1 loci 1414:1417 the FGI loci 1406:1417 the FGI loci 1406:1417 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 7 25 theme flagellin 1351:1359 arg1 island 1371:1376 flagellin methylase island 1351:1376 These flagellin methylase island loci (FMIs) 1345:1388 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 0 26 theme flagellin 17:25 arg1 modification 27:38 flagellin modification 17:38 flagellin modification 17:38 Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae. 27206480 0 27 dep flagella 7:14 arg1 versatile 72:80 versatile 72:80 versatile 72:80 Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae. 27206480 0 27 dep flagella 7:14 arg1 common 54:59 common 54:59 common 54:59 Flashy flagella: flagellin modification is relatively common and highly versatile among the Enterobacteriaceae. 27206480 1 28 gly glycosylation 142:154 arg1 protein 173:179 the flagellin protein 159:179 the flagellin protein 159:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 3 29 theme representative 616:629 arg1 genomes 631:637 2,000 representative genomes 610:637 2,000 representative genomes of Enterobacteriaceae 610:659 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 5 30 theme glycan 1030:1035 arg1 loci 982:985 These loci 976:985 These loci 976:985 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 5 30 theme glycan 1030:1035 arg1 structures 1037:1046 highly variable flagellin glycan structures 1004:1046 highly variable flagellin glycan structures 1004:1046 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 5 31 theme variable 1011:1018 arg1 loci 982:985 These loci 976:985 These loci 976:985 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 5 31 theme variable 1011:1018 arg1 structures 1037:1046 highly variable flagellin glycan structures 1004:1046 highly variable flagellin glycan structures 1004:1046 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 8 32 theme important 1617:1625 arg1 roles 1638:1642 important biological roles 1617:1642 important biological roles 1617:1642 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 5 33 theme lipoyl 1129:1134 arg1 groups 1146:1151 formyl, methyl, acetyl, lipoyl and amino groups 1105:1151 groups 1146:1151 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 1 34 theme BACKGROUND 112:121 arg1 glycosylation 142:154 BACKGROUND Post-translational glycosylation 112:154 BACKGROUND Post-translational glycosylation of the flagellin protein 112:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 5 35 theme formyl 1105:1110 arg1 groups 1146:1151 formyl, methyl, acetyl, lipoyl and amino groups 1105:1151 groups 1146:1151 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 2 36 theme genetic 446:452 arg1 characterization 454:469 extensive physiological and genetic characterization 418:469 extensive physiological and genetic characterization 418:469 However to date, despite extensive physiological and genetic characterization, it has never been reported for the peritrichously flagellate Enterobacteriaceae. 27206480 1 37 theme biofilm 322:328 arg1 formation 330:338 biofilm formation 322:338 biofilm formation 322:338 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 7 38 theme methylase 1361:1369 arg1 island 1371:1376 flagellin methylase island 1351:1376 These flagellin methylase island loci (FMIs) 1345:1388 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 4 39 theme mobile 876:881 arg1 elements 891:898 mobile genetic elements 876:898 mobile genetic elements 876:898 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 6 40 theme Salmonella 1256:1265 arg1 pathogen 1247:1254 the enterobacterial pathogen 1227:1254 the enterobacterial pathogen Salmonella enterica 1227:1274 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 6 41 theme several 1304:1310 arg1 taxa 1321:1324 several distinct taxa 1304:1324 several distinct taxa within the family 1304:1342 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 5 42 theme amino 1140:1144 arg1 groups 1146:1151 formyl, methyl, acetyl, lipoyl and amino groups 1105:1151 groups 1146:1151 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 1 43 attach linked 245:250 arg2 glycosylation 142:154 BACKGROUND Post-translational glycosylation 112:154 BACKGROUND Post-translational glycosylation of the flagellin protein 112:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 virulence 382:390 virulence 382:390 virulence 382:390 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 phenotypes 263:272 several phenotypes 255:272 several phenotypes 255:272 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 motility 312:319 motility 312:319 motility 312:319 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 evasion 353:359 host immune evasion 341:359 host immune evasion 341:359 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 formation 330:338 biofilm formation 322:338 biofilm formation 322:338 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 43 attach linked 245:250 arg1 biosynthesis 295:306 flagellar biosynthesis 285:306 flagellar biosynthesis 285:306 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 2 44 theme physiological 428:440 arg1 characterization 454:469 extensive physiological and genetic characterization 418:469 extensive physiological and genetic characterization 418:469 However to date, despite extensive physiological and genetic characterization, it has never been reported for the peritrichously flagellate Enterobacteriaceae. 27206480 8 45 theme biological 1627:1636 arg1 roles 1638:1642 important biological roles 1617:1642 important biological roles 1617:1642 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 4 46 theme FGI 966:968 arg1 loci 970:973 the FGI loci 962:973 the FGI loci 962:973 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 47 theme elements 891:898 arg1 Differences 784:794 Differences 784:794 Differences in the G + C content of the FGIs and the rest of the genome 784:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 47 theme elements 891:898 arg1 presence 864:871 the presence 860:871 the presence of mobile genetic elements 860:898 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 1 48 theme Post-translational 123:140 arg1 glycosylation 142:154 BACKGROUND Post-translational glycosylation 112:154 BACKGROUND Post-translational glycosylation of the flagellin protein 112:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 49 theme host 341:344 arg1 evasion 353:359 host immune evasion 341:359 host immune evasion 341:359 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 8 50 dep CONCLUSIONS 1482:1492 arg1 suggests 1598:1605 suggests 1598:1605 suggests they 1598:1610 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 5 51 theme methyl 1113:1118 arg1 groups 1146:1151 formyl, methyl, acetyl, lipoyl and amino groups 1105:1151 groups 1146:1151 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 4 52 theme genetic 883:889 arg1 elements 891:898 mobile genetic elements 876:898 mobile genetic elements 876:898 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 6 53 dep Salmonella 1256:1265 arg1 enterica 1267:1274 enterica 1267:1274 enterica 1267:1274 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 1 54 theme immune 346:351 arg1 evasion 353:359 host immune evasion 341:359 host immune evasion 341:359 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 8 55 dep loci 1551:1554 arg1 both 1557:1560 both 1557:1560 both 1557:1560 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 27206480 5 56 theme sugar 1063:1067 arg1 backbones 1069:1077 distinct sugar backbones 1054:1077 distinct sugar backbones 1054:1077 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 3 57 dep analyzed 601:608 arg1 Using 561:565 Using 561:565 Using comparative genomic approaches 561:596 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 5 58 theme flagellin 1020:1028 arg1 loci 982:985 These loci 976:985 These loci 976:985 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 5 58 theme flagellin 1020:1028 arg1 structures 1037:1046 highly variable flagellin glycan structures 1004:1046 highly variable flagellin glycan structures 1004:1046 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 6 59 theme N-lysine 1171:1178 arg1 methylase 1180:1188 an N-lysine methylase 1168:1188 an N-lysine methylase 1168:1188 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 6 59 theme N-lysine 1171:1178 arg1 common 1291:1296 common 1291:1296 common 1291:1296 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 6 59 theme N-lysine 1171:1178 arg1 FliB 1191:1194 FliB 1191:1194 FliB 1191:1194 Additionally, an N-lysine methylase, FliB, previously identified only in the enterobacterial pathogen Salmonella enterica, is relatively common among several distinct taxa within the family. 27206480 5 60 theme acetyl 1121:1126 arg1 groups 1146:1151 formyl, methyl, acetyl, lipoyl and amino groups 1105:1151 groups 1146:1151 These loci therefore encode highly variable flagellin glycan structures, with distinct sugar backbones, heavily substituted with formyl, methyl, acetyl, lipoyl and amino groups. 27206480 4 61 theme genome 849:854 arg1 rest 837:840 the rest 833:840 the rest of the genome 833:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 61 theme genome 849:854 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 62 from presence 864:871 arg1 rest 837:840 the rest 833:840 the rest of the genome 833:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 62 from presence 864:871 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 3 63 theme genomic 579:585 arg1 approaches 587:596 comparative genomic approaches 567:596 comparative genomic approaches 567:596 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 7 64 theme island 1371:1376 arg1 loci 1378:1381 These flagellin methylase island loci 1345:1381 These flagellin methylase island loci (FMIs) 1345:1388 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 7 64 theme island 1371:1376 arg1 FMIs 1384:1387 FMIs 1384:1387 FMIs 1384:1387 These flagellin methylase island loci (FMIs), in contrast to the FGI loci, appear to be stably maintained within these diverse lineages. 27206480 3 65 theme flagellin 676:684 arg1 islands 700:706 flagellin glycosylation islands 676:706 flagellin glycosylation islands 676:706 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 1 66 theme flagellin 163:171 arg1 protein 173:179 the flagellin protein 159:179 the flagellin protein 159:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 virulence 382:390 virulence 382:390 virulence 382:390 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 phenotypes 263:272 several phenotypes 255:272 several phenotypes 255:272 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 motility 312:319 motility 312:319 motility 312:319 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 evasion 353:359 host immune evasion 341:359 host immune evasion 341:359 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 formation 330:338 biofilm formation 322:338 biofilm formation 322:338 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 1 67 theme several 255:261 arg1 biosynthesis 295:306 flagellar biosynthesis 285:306 flagellar biosynthesis 285:306 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 3 68 theme glycosylation 686:698 arg1 islands 700:706 flagellin glycosylation islands 676:706 flagellin glycosylation islands 676:706 RESULTS Using comparative genomic approaches we analyzed 2,000 representative genomes of Enterobacteriaceae, and show that flagellin glycosylation islands are relatively common and extremely versatile among members of this family. 27206480 1 69 theme protein 173:179 arg1 glycosylation 142:154 BACKGROUND Post-translational glycosylation 112:154 BACKGROUND Post-translational glycosylation of the flagellin protein 112:179 BACKGROUND Post-translational glycosylation of the flagellin protein is relatively common among Gram-negative bacteria, and has been linked to several phenotypes, including flagellar biosynthesis and motility, biofilm formation, host immune evasion and manipulation and virulence. 27206480 4 70 from Differences 784:794 arg1 rest 837:840 the rest 833:840 the rest of the genome 833:854 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 4 70 from Differences 784:794 arg1 content 809:815 the G + C content 799:815 the G + C content of the FGIs 799:827 Differences in the G + C content of the FGIs and the rest of the genome and the presence of mobile genetic elements provide evidence of horizontal gene transfer occurring within the FGI loci. 27206480 8 71 dep prevalence 1498:1507 arg1 The 1494:1496 The 1494:1496 The 1494:1496 CONCLUSIONS The prevalence and versatility of flagellin modification loci, both glycosylation and methylation loci, suggests they play important biological roles among the Enterobacteriaceae. 28244758 4 0 theme PTMs 552:555 arg1 PTMs 552:555 the most common and functionally important PTMs 509:555 the most common and functionally important PTMs 509:555 Glycosylation is one of the most common and functionally important PTMs. 28244758 4 0 theme PTMs 552:555 arg1 one 502:504 one 502:504 one 502:504 Glycosylation is one of the most common and functionally important PTMs. 28244758 5 1 theme protein 622:628 arg1 folding 630:636 protein folding 622:636 protein folding 622:636 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 1 2 theme genomic 159:165 arg1 approaches 167:176 advanced proteomic and genomic approaches 136:176 advanced proteomic and genomic approaches 136:176 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 4 3 theme important 542:550 arg1 PTMs 552:555 the most common and functionally important PTMs 509:555 the most common and functionally important PTMs 509:555 Glycosylation is one of the most common and functionally important PTMs. 28244758 2 4 theme protein 288:294 arg1 component 296:304 the protein component 284:304 the protein component 284:304 Most candidate biomarkers are focused on the protein component. 28244758 3 5 theme protein 377:383 arg1 sequences 385:393 protein sequences 377:393 protein sequences 377:393 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 3 6 with information 428:438 arg1 regards 445:451 regards 445:451 regards to sensitivity and specificity 445:482 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 6 7 theme various 766:772 arg1 types 774:778 various types 766:778 various types of cancer 766:788 Cancer-associated aberrant glycosylation has been identified in various types of cancer. 28244758 8 8 dep proteins 956:963 arg1 LIFR 966:969 LIFR 966:969 LIFR 966:969 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 8 dep proteins 956:963 arg1 HPT 986:988 HPT 986:988 HPT 986:988 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 8 dep proteins 956:963 arg1 VP13A 979:983 VP13A 979:983 VP13A 979:983 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 8 dep proteins 956:963 arg1 CE350 972:976 CE350 972:976 CE350 972:976 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 9 dep structures 1062:1071 arg1 compared 1076:1083 compared 1076:1083 compared to those of controls 1076:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 5 10 theme host-pathogen 639:651 arg1 interactions 653:664 host-pathogen interactions 639:664 host-pathogen interactions 639:664 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 4 11 theme common 518:523 arg1 PTMs 552:555 the most common and functionally important PTMs 509:555 the most common and functionally important PTMs 509:555 Glycosylation is one of the most common and functionally important PTMs. 28244758 5 12 theme biological 590:599 arg1 folding 630:636 protein folding 622:636 protein folding 622:636 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 12 theme biological 590:599 arg1 interactions 653:664 host-pathogen interactions 639:664 host-pathogen interactions 639:664 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 12 theme biological 590:599 arg1 response 674:681 immune response 667:681 immune response 667:681 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 12 theme biological 590:599 arg1 inflammation 688:699 inflammation 688:699 inflammation 688:699 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 12 theme biological 590:599 arg1 processes 601:609 many biological processes 585:609 many biological processes 585:609 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 3 13 theme superior 408:415 arg1 information 428:438 superior diagnostic information 408:438 superior diagnostic information with regards to sensitivity and specificity 408:482 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 7 14 theme tumors 927:932 arg1 detection 914:922 potentially specific detection 893:922 potentially specific detection of tumors 893:932 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 7 14 theme tumors 927:932 arg1 opportunity 861:871 an excellent opportunity 848:871 an excellent opportunity for diagnostics 848:887 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 5 15 theme immune 667:672 arg1 response 674:681 immune response 667:681 immune response 667:681 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 0 16 from Changes 14:20 arg1 Proteins 31:38 Serum Proteins 25:38 Serum Proteins 25:38 Glycosylation Changes in Serum Proteins Identify Patients with Pancreatic Cancer. 28244758 1 17 theme few 184:186 arg1 biomarkers 188:197 very few biomarkers 179:197 very few biomarkers 179:197 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 0 18 theme Serum 25:29 arg1 Proteins 31:38 Serum Proteins 25:38 Serum Proteins 25:38 Glycosylation Changes in Serum Proteins Identify Patients with Pancreatic Cancer. 28244758 8 19 theme cancer 1021:1026 arg1 patients 1028:1035 pancreatic cancer patients 1010:1035 pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 1010:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 20 theme pancreatic 1010:1019 arg1 patients 1028:1035 pancreatic cancer patients 1010:1035 pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 1010:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 6 21 theme cancer 783:788 arg1 types 774:778 various types 766:778 various types of cancer 766:788 Cancer-associated aberrant glycosylation has been identified in various types of cancer. 28244758 6 22 gly glycosylation 729:741 arg1 types 774:778 various types 766:778 various types of cancer 766:788 Cancer-associated aberrant glycosylation has been identified in various types of cancer. 28244758 7 23 theme specific 905:912 arg1 detection 914:922 potentially specific detection 893:922 potentially specific detection of tumors 893:932 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 6 24 theme aberrant 720:727 arg1 glycosylation 729:741 Cancer-associated aberrant glycosylation 702:741 Cancer-associated aberrant glycosylation 702:741 Cancer-associated aberrant glycosylation has been identified in various types of cancer. 28244758 6 25 theme Cancer-associated 702:718 arg1 glycosylation 729:741 Cancer-associated aberrant glycosylation 702:741 Cancer-associated aberrant glycosylation 702:741 Cancer-associated aberrant glycosylation has been identified in various types of cancer. 28244758 3 26 with combination 360:370 arg1 sequences 385:393 protein sequences 377:393 protein sequences 377:393 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 5 27 theme central 569:575 arg1 role 577:580 a central role 567:580 a central role 567:580 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 3 28 theme diagnostic 417:426 arg1 information 428:438 superior diagnostic information 408:438 superior diagnostic information with regards to sensitivity and specificity 408:482 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 7 29 theme excellent 851:859 arg1 opportunity 861:871 an excellent opportunity 848:871 an excellent opportunity for diagnostics 848:887 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 8 30 from patients 1028:1035 arg1 sera 1000:1003 sera 1000:1003 sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 1000:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 7 31 theme epitopes 828:835 arg1 Expression 791:800 Expression 791:800 Expression of cancer-specific glycan epitopes 791:835 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 3 32 theme post-translational 317:334 arg1 PTMs 351:354 PTMs 351:354 PTMs 351:354 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 3 32 theme post-translational 317:334 arg1 modifications 336:348 post-translational modifications 317:348 post-translational modifications (PTMs) 317:355 Targeting post-translational modifications (PTMs) in combination with protein sequences will provide superior diagnostic information with regards to sensitivity and specificity. 28244758 1 33 theme biomarker 110:118 arg1 discovery 120:128 biomarker discovery 110:128 biomarker discovery 110:128 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 8 34 theme glycan 1055:1060 arg1 structures 1062:1071 aberrant glycan structures 1046:1071 aberrant glycan structures as compared to those of controls 1046:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 0 35 theme Glycosylation 0:12 arg1 Changes 14:20 Glycosylation Changes 0:20 Glycosylation Changes in Serum Proteins 0:38 Glycosylation Changes in Serum Proteins Identify Patients with Pancreatic Cancer. 28244758 8 36 theme aberrant 1046:1053 arg1 structures 1062:1071 aberrant glycan structures 1046:1071 aberrant glycan structures as compared to those of controls 1046:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 5 37 theme many 585:588 arg1 folding 630:636 protein folding 622:636 protein folding 622:636 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 37 theme many 585:588 arg1 interactions 653:664 host-pathogen interactions 639:664 host-pathogen interactions 639:664 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 37 theme many 585:588 arg1 response 674:681 immune response 667:681 immune response 667:681 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 37 theme many 585:588 arg1 inflammation 688:699 inflammation 688:699 inflammation 688:699 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 5 37 theme many 585:588 arg1 processes 601:609 many biological processes 585:609 many biological processes 585:609 It plays a central role in many biological processes, including protein folding, host-pathogen interactions, immune response, and inflammation. 28244758 1 38 theme discovery 120:128 arg1 decade 100:105 a decade 98:105 a decade of biomarker discovery 98:128 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 7 39 theme cancer-specific 805:819 arg1 epitopes 828:835 cancer-specific glycan epitopes 805:835 cancer-specific glycan epitopes 805:835 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 1 40 theme clinical 221:228 arg1 diagnostics 230:240 clinical diagnostics 221:240 clinical diagnostics 221:240 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 7 41 theme glycan 821:826 arg1 epitopes 828:835 cancer-specific glycan epitopes 805:835 cancer-specific glycan epitopes 805:835 Expression of cancer-specific glycan epitopes represents an excellent opportunity for diagnostics and potentially specific detection of tumors. 28244758 2 42 theme candidate 248:256 arg1 biomarkers 258:267 Most candidate biomarkers 243:267 Most candidate biomarkers 243:267 Most candidate biomarkers are focused on the protein component. 28244758 0 43 with Patients 49:56 arg1 Cancer 74:79 Pancreatic Cancer 63:79 Pancreatic Cancer 63:79 Glycosylation Changes in Serum Proteins Identify Patients with Pancreatic Cancer. 28244758 1 44 theme advanced 136:143 arg1 approaches 167:176 advanced proteomic and genomic approaches 136:176 advanced proteomic and genomic approaches 136:176 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 8 45 contain carrying 1037:1044 arg2 structures 1062:1071 aberrant glycan structures 1046:1071 aberrant glycan structures as compared to those of controls 1046:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 45 contain carrying 1037:1044 arg1 patients 1028:1035 pancreatic cancer patients 1010:1035 pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 1010:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 0 46 theme Pancreatic 63:72 arg1 Cancer 74:79 Pancreatic Cancer 63:79 Pancreatic Cancer 63:79 Glycosylation Changes in Serum Proteins Identify Patients with Pancreatic Cancer. 28244758 2 47 theme Most 243:246 arg1 biomarkers 258:267 Most candidate biomarkers 243:267 Most candidate biomarkers 243:267 Most candidate biomarkers are focused on the protein component. 28244758 1 48 theme proteomic 145:153 arg1 approaches 167:176 advanced proteomic and genomic approaches 136:176 advanced proteomic and genomic approaches 136:176 After more than a decade of biomarker discovery using advanced proteomic and genomic approaches, very few biomarkers have been involved in clinical diagnostics. 28244758 8 49 located found 991:995 arg2 proteins 956:963 four proteins 951:963 four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 951:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 28244758 8 49 located found 991:995 arg1 sera 1000:1003 sera 1000:1003 sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls 1000:1104 Here, we report four proteins (LIFR, CE350, VP13A, HPT) found in sera from pancreatic cancer patients carrying aberrant glycan structures as compared to those of controls. 24352591 8 0 theme expressed 1785:1793 arg1 phosphacan 1795:1804 exogenously expressed phosphacan 1773:1804 exogenously expressed phosphacan 1773:1804 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 3 1 from phosphacan 733:742 arg1 brains 764:769 developing mouse brains 747:769 developing mouse brains 747:769 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 1 2 link O-mannose-linked 290:305 arg1 glycans 307:313 N-linked and O-mannose-linked glycans 277:313 N-linked and O-mannose-linked glycans in the nervous system 277:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 7 3 theme monoclonal 1593:1602 arg1 mAb 1614:1616 mAb 1614:1616 mAb 1614:1616 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 7 3 theme monoclonal 1593:1602 arg1 antibody 1604:1611 the 6B4 monoclonal antibody 1585:1611 the 6B4 monoclonal antibody (mAb) 1585:1617 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 3 4 gly glycoprotein 695:706 arg1 glycoprotein 695:706 the major carrier glycoprotein 677:706 the major carrier glycoprotein of the O-linked HNK-1 677:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 1 5 link N-linked 277:284 arg1 glycans 307:313 N-linked and O-mannose-linked glycans 277:313 N-linked and O-mannose-linked glycans in the nervous system 277:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 8 6 from HNK-1 1951:1955 arg1 phosphacan 1960:1969 phosphacan 1960:1969 phosphacan 1960:1969 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 9 7 from HNK-1 2043:2047 arg1 brain 2056:2060 the brain 2052:2060 the brain 2052:2060 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 3 8 theme O-glycan 796:803 arg1 structures 805:814 the major O-glycan structures 786:814 the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan 786:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 1 9 from glycans 307:313 arg1 system 330:335 the nervous system 318:335 the nervous system 318:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 6 10 theme O-linked 1472:1479 arg1 HNK-1 1481:1485 O-linked HNK-1 1472:1485 O-linked HNK-1 onto phosphacan using cultured cells 1472:1522 Consistent with this result, we revealed that GlcAT-P specifically synthesized O-linked HNK-1 onto phosphacan using cultured cells. 24352591 4 11 from epitope 905:911 arg1 phosphacan 916:925 phosphacan 916:925 phosphacan 916:925 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 12 theme protein 969:975 arg1 β1,2-N-acetylglucosaminyltransferase 987:1022 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 12 theme protein 969:975 arg1 N-acetylglucosaminyltransferase 1030:1060 an N-acetylglucosaminyltransferase 1027:1060 an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis 1027:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 3 13 theme terminal 827:834 arg1 epitope 842:848 the terminal HNK-1 epitope 823:848 the terminal HNK-1 epitope from partially purified phosphacan 823:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 3 14 theme spectrometric 639:651 arg1 analyses 653:660 mass spectrometric analyses 634:660 mass spectrometric analyses 634:660 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 4 15 theme β1,2-N-acetylglucosaminyltransferase 987:1022 arg1 knockout 957:964 knockout 957:964 knockout 957:964 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 3 16 theme major 681:685 arg1 glycoprotein 695:706 the major carrier glycoprotein 677:706 the major carrier glycoprotein of the O-linked HNK-1 677:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 1 17 theme human 150:154 arg1 carbohydrate 181:192 The human natural killer-1 (HNK-1) carbohydrate 146:192 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) 146:259 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 2 18 link O-mannose-linked 556:571 arg1 epitope 579:585 the O-mannose-linked HNK-1 epitope 552:585 the O-mannose-linked HNK-1 epitope 552:585 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 19 theme killer-1 78:85 arg1 glycan 87:92 O-mannose-linked human natural killer-1 glycan 47:92 O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains 47:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 1 20 theme killer-1 164:171 arg1 carbohydrate 181:192 The human natural killer-1 (HNK-1) carbohydrate 146:192 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) 146:259 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 1 21 theme HNK-1 174:178 arg1 carbohydrate 181:192 The human natural killer-1 (HNK-1) carbohydrate 146:192 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) 146:259 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 4 22 link O-linked 1156:1163 arg1 HNK-1 1165:1169 the O-linked HNK-1 1152:1169 the O-linked HNK-1 1152:1169 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 2 23 theme HNK-1 573:577 arg1 epitope 579:585 the O-mannose-linked HNK-1 epitope 552:585 the O-mannose-linked HNK-1 epitope 552:585 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 1 24 dep learning 353:360 arg1 functions 373:381 functions 373:381 functions 373:381 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 2 25 gly glycoproteins 434:446 arg1 glycoproteins 434:446 carrier glycoproteins 426:446 carrier glycoproteins 426:446 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 26 theme developing 121:130 arg1 brains 138:143 developing mouse brains 121:143 developing mouse brains 121:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 4 27 theme O-linked 1156:1163 arg1 HNK-1 1165:1169 the O-linked HNK-1 1152:1169 the O-linked HNK-1 1152:1169 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 2 28 theme carrier 426:432 arg1 glycoproteins 434:446 carrier glycoproteins 426:446 carrier glycoproteins 426:446 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 29 theme Structural 0:9 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 0:144 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 5 30 theme gene-deficient 1306:1319 arg1 mice 1321:1324 the gene-deficient mice 1302:1324 the gene-deficient mice of GlcAT-P 1302:1335 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 30 theme gene-deficient 1306:1319 arg1 one 1338:1340 one 1338:1340 one 1338:1340 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 30 theme gene-deficient 1306:1319 arg1 glucuronyltransferases 1349:1370 the glucuronyltransferases 1345:1370 the glucuronyltransferases for HNK-1 synthesis 1345:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 9 31 theme first 1984:1988 arg1 study 1990:1994 the first study 1980:1994 the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan 1980:2092 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 9 31 theme first 1984:1988 arg1 This 1972:1975 This 1972:1975 This 1972:1975 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 0 32 theme biochemical 15:25 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 0:144 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 1 33 theme nervous 322:328 arg1 system 330:335 the nervous system 318:335 the nervous system 318:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 4 34 theme reducing 1131:1138 arg1 terminal 1140:1147 the reducing terminal 1127:1147 the reducing terminal of the O-linked HNK-1 1127:1169 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 34 theme reducing 1131:1138 arg1 mannose 1174:1180 mannose 1174:1180 mannose 1174:1180 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 8 35 theme 6B4 1704:1706 arg1 mAb 1708:1710 the 6B4 mAb 1700:1710 the 6B4 mAb 1700:1710 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 4 36 theme O-linked 890:897 arg1 epitope 905:911 The O-linked HNK-1 epitope 886:911 The O-linked HNK-1 epitope on phosphacan 886:925 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 2 37 theme N-linked 456:463 arg1 epitope 471:477 the N-linked HNK-1 epitope 452:477 the N-linked HNK-1 epitope 452:477 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 5 38 theme O-mannose-linked 1273:1288 arg1 HNK-1 1290:1294 O-mannose-linked HNK-1 1273:1294 O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis 1273:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 0 39 theme human 64:68 arg1 glycan 87:92 O-mannose-linked human natural killer-1 glycan 47:92 O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains 47:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 7 40 theme unique 1653:1658 arg1 glycoform 1671:1679 a unique phosphacan glycoform 1651:1679 a unique phosphacan glycoform 1651:1679 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 2 41 gly glycoproteins 506:518 arg1 glycoproteins 506:518 carrier glycoproteins 498:518 carrier glycoproteins 498:518 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 5 42 link O-mannose-linked 1273:1288 arg1 HNK-1 1290:1294 O-mannose-linked HNK-1 1273:1294 O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis 1273:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 3 43 link O-linked 715:722 arg1 HNK-1 724:728 the O-linked HNK-1 711:728 the O-linked HNK-1 711:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 3 44 from phosphacan 874:883 arg1 epitope 842:848 the terminal HNK-1 epitope 823:848 the terminal HNK-1 epitope from partially purified phosphacan 823:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 5 45 theme glucuronyltransferases 1349:1370 arg1 mice 1321:1324 the gene-deficient mice 1302:1324 the gene-deficient mice of GlcAT-P 1302:1335 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 45 theme glucuronyltransferases 1349:1370 arg1 one 1338:1340 one 1338:1340 one 1338:1340 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 45 theme glucuronyltransferases 1349:1370 arg1 glucuronyltransferases 1349:1370 the glucuronyltransferases 1345:1370 the glucuronyltransferases for HNK-1 synthesis 1345:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 8 46 theme GlcAT-P-deficient 1745:1761 arg1 mice 1763:1766 GlcAT-P-deficient mice 1745:1766 GlcAT-P-deficient mice 1745:1766 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 3 47 theme HNK-1 724:728 arg1 glycoprotein 695:706 the major carrier glycoprotein 677:706 the major carrier glycoprotein of the O-linked HNK-1 677:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 4 48 theme glycan 1093:1098 arg1 synthesis 1100:1108 O-mannose-linked glycan synthesis 1076:1108 O-mannose-linked glycan synthesis 1076:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 5 49 theme HNK-1 1376:1380 arg1 synthesis 1382:1390 HNK-1 synthesis 1376:1390 HNK-1 synthesis 1376:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 8 50 link O-mannose-linked 1934:1949 arg1 HNK-1 1951:1955 O-mannose-linked HNK-1 1934:1955 O-mannose-linked HNK-1 on phosphacan 1934:1969 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 3 51 theme purified 865:872 arg1 phosphacan 874:883 partially purified phosphacan 855:883 partially purified phosphacan 855:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 2 52 theme whole/core 393:402 arg1 structures 411:420 whole/core glycan structures 393:420 whole/core glycan structures 393:420 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 8 53 theme 6B4 1840:1842 arg1 mAb 1844:1846 the 6B4 mAb 1836:1846 the 6B4 mAb 1836:1846 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 1 54 theme N-linked 277:284 arg1 glycans 307:313 N-linked and O-mannose-linked glycans 277:313 N-linked and O-mannose-linked glycans in the nervous system 277:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 3 55 theme developing 747:756 arg1 brains 764:769 developing mouse brains 747:769 developing mouse brains 747:769 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 1 56 theme O-mannose-linked 290:305 arg1 glycans 307:313 N-linked and O-mannose-linked glycans 277:313 N-linked and O-mannose-linked glycans in the nervous system 277:335 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 7 57 theme antibody 1604:1611 arg1 epitope 1574:1580 the as-yet-unknown epitope 1555:1580 the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform 1555:1679 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 3 58 theme major 790:794 arg1 structures 805:814 the major O-glycan structures 786:814 the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan 786:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 9 59 theme 6B4 2009:2011 arg1 HNK-1 2043:2047 6B4 mAb-reactive O-mannose-linked HNK-1 2009:2047 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain 2009:2060 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 7 60 theme 6B4 1589:1591 arg1 mAb 1614:1616 mAb 1614:1616 mAb 1614:1616 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 7 60 theme 6B4 1589:1591 arg1 antibody 1604:1611 the 6B4 monoclonal antibody 1585:1611 the 6B4 monoclonal antibody (mAb) 1585:1617 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 0 61 link O-mannose-linked 47:62 arg1 glycan 87:92 O-mannose-linked human natural killer-1 glycan 47:92 O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains 47:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 1 62 theme sulfated 207:214 arg1 trisaccharide 216:228 a sulfated trisaccharide 205:228 a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) 205:259 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 1 62 theme sulfated 207:214 arg1 HSO3-3GlcAβ1-3Galβ1-4GlcNAc- 231:258 HSO3-3GlcAβ1-3Galβ1-4GlcNAc- 231:258 HSO3-3GlcAβ1-3Galβ1-4GlcNAc- 231:258 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 7 63 theme as-yet-unknown 1559:1572 arg1 epitope 1574:1580 the as-yet-unknown epitope 1555:1580 the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform 1555:1679 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 8 64 theme 6B4 1900:1902 arg1 mAb 1904:1906 the 6B4 mAb 1896:1906 the 6B4 mAb 1896:1906 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 9 65 theme mAb-reactive 2013:2024 arg1 HNK-1 2043:2047 6B4 mAb-reactive O-mannose-linked HNK-1 2009:2047 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain 2009:2060 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 3 66 theme mass 634:637 arg1 analyses 653:660 mass spectrometric analyses 634:660 mass spectrometric analyses 634:660 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 3 67 theme HNK-1 836:840 arg1 epitope 842:848 the terminal HNK-1 epitope 823:848 the terminal HNK-1 epitope from partially purified phosphacan 823:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 8 68 theme O-mannose-linked 1934:1949 arg1 HNK-1 1951:1955 O-mannose-linked HNK-1 1934:1955 O-mannose-linked HNK-1 on phosphacan 1934:1969 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 4 69 theme O-mannose 977:985 arg1 β1,2-N-acetylglucosaminyltransferase 987:1022 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 69 theme O-mannose 977:985 arg1 N-acetylglucosaminyltransferase 1030:1060 an N-acetylglucosaminyltransferase 1027:1060 an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis 1027:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 0 70 theme glycan 87:92 arg1 characterization 27:42 Structural and biochemical characterization 0:42 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 0:144 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 1 71 theme natural 156:162 arg1 carbohydrate 181:192 The human natural killer-1 (HNK-1) carbohydrate 146:192 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) 146:259 The human natural killer-1 (HNK-1) carbohydrate comprising a sulfated trisaccharide (HSO3-3GlcAβ1-3Galβ1-4GlcNAc-) is expressed on N-linked and O-mannose-linked glycans in the nervous system and involved in learning and memory functions. 24352591 9 72 theme O-mannose-linked 2026:2041 arg1 HNK-1 2043:2047 6B4 mAb-reactive O-mannose-linked HNK-1 2009:2047 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain 2009:2060 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 4 73 theme essential 1062:1070 arg1 β1,2-N-acetylglucosaminyltransferase 987:1022 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 969:1024 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 73 theme essential 1062:1070 arg1 N-acetylglucosaminyltransferase 1030:1060 an N-acetylglucosaminyltransferase 1027:1060 an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis 1027:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 2 74 theme biosynthetic 528:539 arg1 pathway 541:547 the biosynthetic pathway 524:547 the biosynthetic pathway 524:547 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 7 75 gly glycoform 1671:1679 arg1 phosphacan 1660:1669 a unique phosphacan glycoform 1651:1679 a unique phosphacan glycoform 1651:1679 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 2 76 theme epitope 579:585 arg1 glycoproteins 506:518 carrier glycoproteins 498:518 carrier glycoproteins 498:518 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 2 76 theme epitope 579:585 arg1 pathway 541:547 the biosynthetic pathway 524:547 the biosynthetic pathway 524:547 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 77 theme mouse 132:136 arg1 brains 138:143 developing mouse brains 121:143 developing mouse brains 121:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 4 78 theme O-mannose-linked 1076:1091 arg1 synthesis 1100:1108 O-mannose-linked glycan synthesis 1076:1108 O-mannose-linked glycan synthesis 1076:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 6 79 theme cultured 1509:1516 arg1 cells 1518:1522 cultured cells 1509:1522 cultured cells 1509:1522 Consistent with this result, we revealed that GlcAT-P specifically synthesized O-linked HNK-1 onto phosphacan using cultured cells. 24352591 4 80 link O-linked 890:897 arg1 epitope 905:911 The O-linked HNK-1 epitope 886:911 The O-linked HNK-1 epitope on phosphacan 886:925 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 81 theme HNK-1 1165:1169 arg1 terminal 1140:1147 the reducing terminal 1127:1147 the reducing terminal of the O-linked HNK-1 1127:1169 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 4 81 theme HNK-1 1165:1169 arg1 mannose 1174:1180 mannose 1174:1180 mannose 1174:1180 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 8 82 theme mAb 1708:1710 arg1 reactivity 1686:1695 The reactivity 1682:1695 The reactivity of the 6B4 mAb 1682:1710 The reactivity of the 6B4 mAb almost completely disappeared in GlcAT-P-deficient mice, and exogenously expressed phosphacan was selectively recognized by the 6B4 mAb when co-expressed with GlcAT-P, suggesting that the 6B4 mAb preferentially recognizes O-mannose-linked HNK-1 on phosphacan. 24352591 3 83 contain having 816:821 arg2 epitope 842:848 the terminal HNK-1 epitope 823:848 the terminal HNK-1 epitope from partially purified phosphacan 823:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 3 83 contain having 816:821 arg1 structures 805:814 the major O-glycan structures 786:814 the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan 786:883 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 2 84 theme HNK-1 465:469 arg1 epitope 471:477 the N-linked HNK-1 epitope 452:477 the N-linked HNK-1 epitope 452:477 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 85 theme O-mannose-linked 47:62 arg1 glycan 87:92 O-mannose-linked human natural killer-1 glycan 47:92 O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains 47:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 5 86 theme GlcAT-P 1329:1335 arg1 mice 1321:1324 the gene-deficient mice 1302:1324 the gene-deficient mice of GlcAT-P 1302:1335 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 86 theme GlcAT-P 1329:1335 arg1 one 1338:1340 one 1338:1340 one 1338:1340 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 5 86 theme GlcAT-P 1329:1335 arg1 glucuronyltransferases 1349:1370 the glucuronyltransferases 1345:1370 the glucuronyltransferases for HNK-1 synthesis 1345:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 3 87 theme carrier 687:693 arg1 glycoprotein 695:706 the major carrier glycoprotein 677:706 the major carrier glycoprotein of the O-linked HNK-1 677:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 2 88 theme O-mannose-linked 556:571 arg1 epitope 579:585 the O-mannose-linked HNK-1 epitope 552:585 the O-mannose-linked HNK-1 epitope 552:585 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 0 89 theme natural 70:76 arg1 glycan 87:92 O-mannose-linked human natural killer-1 glycan 47:92 O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains 47:143 Structural and biochemical characterization of O-mannose-linked human natural killer-1 glycan expressed on phosphacan in developing mouse brains. 24352591 5 90 theme HNK-1 1290:1294 arg1 biosynthesis 1257:1268 the biosynthesis 1253:1268 the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis 1253:1390 We also showed that glucuronyltransferase-P (GlcAT-P) was involved in the biosynthesis of O-mannose-linked HNK-1 using the gene-deficient mice of GlcAT-P, one of the glucuronyltransferases for HNK-1 synthesis. 24352591 2 91 theme carrier 498:504 arg1 glycoproteins 506:518 carrier glycoproteins 498:518 carrier glycoproteins 498:518 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 4 92 link O-mannose-linked 1076:1091 arg1 synthesis 1100:1108 O-mannose-linked glycan synthesis 1076:1108 O-mannose-linked glycan synthesis 1076:1108 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 2 93 link N-linked 456:463 arg1 epitope 471:477 the N-linked HNK-1 epitope 452:477 the N-linked HNK-1 epitope 452:477 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 4 94 theme HNK-1 899:903 arg1 epitope 905:911 The O-linked HNK-1 epitope 886:911 The O-linked HNK-1 epitope on phosphacan 886:925 The O-linked HNK-1 epitope on phosphacan almost disappeared due to the knockout of protein O-mannose β1,2-N-acetylglucosaminyltransferase 1, an N-acetylglucosaminyltransferase essential for O-mannose-linked glycan synthesis, indicating that the reducing terminal of the O-linked HNK-1 is mannose. 24352591 7 95 theme phosphacan 1660:1669 arg1 glycoform 1671:1679 a unique phosphacan glycoform 1651:1679 a unique phosphacan glycoform 1651:1679 Furthermore, we characterized the as-yet-unknown epitope of the 6B4 monoclonal antibody (mAb), which was thought to recognize a unique phosphacan glycoform. 24352591 6 96 link O-linked 1472:1479 arg1 HNK-1 1481:1485 O-linked HNK-1 1472:1485 O-linked HNK-1 onto phosphacan using cultured cells 1472:1522 Consistent with this result, we revealed that GlcAT-P specifically synthesized O-linked HNK-1 onto phosphacan using cultured cells. 24352591 3 97 theme O-linked 715:722 arg1 HNK-1 724:728 the O-linked HNK-1 711:728 the O-linked HNK-1 711:728 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 9 98 link O-mannose-linked 2026:2041 arg1 HNK-1 2043:2047 6B4 mAb-reactive O-mannose-linked HNK-1 2009:2047 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain 2009:2060 This is the first study to show that 6B4 mAb-reactive O-mannose-linked HNK-1 in the brain is mainly carried by phosphacan. 24352591 3 99 theme mouse 758:762 arg1 brains 764:769 developing mouse brains 747:769 developing mouse brains 747:769 Here, using mass spectrometric analyses, we identified the major carrier glycoprotein of the O-linked HNK-1 as phosphacan in developing mouse brains and determined the major O-glycan structures having the terminal HNK-1 epitope from partially purified phosphacan. 24352591 2 100 theme glycan 404:409 arg1 structures 411:420 whole/core glycan structures 393:420 whole/core glycan structures 393:420 Although whole/core glycan structures and carrier glycoproteins for the N-linked HNK-1 epitope have been studied, carrier glycoproteins and the biosynthetic pathway of the O-mannose-linked HNK-1 epitope have not been fully characterized. 24352591 6 101 with Consistent 1393:1402 arg1 result 1414:1419 this result 1409:1419 this result 1409:1419 Consistent with this result, we revealed that GlcAT-P specifically synthesized O-linked HNK-1 onto phosphacan using cultured cells. 26762173 1 0 theme cellobiohydrolase 147:163 arg1 CBHI 168:171 CBHI 168:171 CBHI 168:171 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 1 0 theme cellobiohydrolase 147:163 arg1 I 165:165 cellobiohydrolase I 147:165 cellobiohydrolase I (CBHI) 147:172 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 1 theme O-glycan/cellulase 851:868 arg1 engineering 870:880 directed O-glycan/cellulase engineering 842:880 directed O-glycan/cellulase engineering 842:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 1 2 theme protein 219:225 arg1 % 210:210 approximately 40% 194:210 approximately 40% of the protein secreted by cells growing in the presence of cellulose 194:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 1 2 theme protein 219:225 arg1 protein 219:225 the protein 215:225 the protein secreted by cells growing in the presence of cellulose 215:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 3 theme genetic 654:660 arg1 resources 662:670 the excellent genetic resources 640:670 the excellent genetic resources available for N. crassa 640:694 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 4 theme fungal 463:468 arg1 O-glycans 470:478 the classic fungal O-glycans 451:478 the classic fungal O-glycans with reducing end mannoses 451:505 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 5 theme N. 619:620 arg1 crassa 622:627 N. crassa 619:627 N. crassa 619:627 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 6 with O-glycans 470:478 arg1 mannoses 498:505 reducing end mannoses 485:505 reducing end mannoses 485:505 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 7 from development 827:837 arg1 evaluation 755:764 the future evaluation 744:764 the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 744:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 8 theme role 773:776 arg1 evaluation 755:764 the future evaluation 744:764 the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 744:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 0 9 from crassa 57:62 arg1 analysis 9:16 O-Glycan analysis 0:16 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 0:63 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 26762173 3 10 theme classic 455:461 arg1 O-glycans 470:478 the classic fungal O-glycans 451:478 the classic fungal O-glycans with reducing end mannoses 451:505 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 1 11 from crassa 140:145 arg1 composition 86:96 the composition 82:96 the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 82:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 12 theme reducing 485:492 arg1 mannoses 498:505 reducing end mannoses 485:505 reducing end mannoses 485:505 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 13 from existence 569:577 arg1 crassa 622:627 N. crassa 619:627 N. crassa 619:627 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 14 theme mannose 413:419 arg1 amounts 402:408 approximately equal amounts 382:408 approximately equal amounts of mannose and galactose 382:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 14 theme mannose 413:419 arg1 galactose 425:433 galactose 425:433 galactose 425:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 14 theme mannose 413:419 arg1 mannose 413:419 mannose 413:419 mannose 413:419 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 3 15 theme N. 686:687 arg1 crassa 689:694 N. crassa 686:694 N. crassa 686:694 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 16 theme O-glycosylation 781:795 arg1 role 773:776 the role 769:776 the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 769:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 17 theme end 494:496 arg1 mannoses 498:505 reducing end mannoses 485:505 reducing end mannoses 485:505 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 18 theme engineering 870:880 arg1 function 810:817 cellulase function 800:817 cellulase function 800:817 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 18 theme engineering 870:880 arg1 development 827:837 the development 823:837 the development of directed O-glycan/cellulase engineering 823:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 0 19 theme O-Glycan 0:7 arg1 analysis 9:16 O-Glycan analysis 0:16 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 0:63 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 26762173 3 20 theme reducing 527:534 arg1 galactoses 540:549 reducing end galactoses 527:549 reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 527:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 21 dep suggest 557:563 arg1 enable 737:742 enable 737:742 may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 733:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 0 22 theme cellobiohydrolase 21:37 arg1 analysis 9:16 O-Glycan analysis 0:16 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 0:63 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 26762173 3 23 theme CBHI 718:721 arg1 O-glycans 723:731 the CBHI O-glycans 714:731 the CBHI O-glycans 714:731 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 24 from role 773:776 arg1 function 810:817 cellulase function 800:817 cellulase function 800:817 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 24 from role 773:776 arg1 development 827:837 the development 823:837 the development of directed O-glycan/cellulase engineering 823:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 1 25 from glycans 114:120 arg1 crassa 140:145 the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 125:280 crassa 140:145 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 26 from function 810:817 arg1 evaluation 755:764 the future evaluation 744:764 the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 744:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 27 with O-glycosylated 291:304 arg1 types 342:346 three types 336:346 three types of branched, O-glycans containing approximately equal amounts of mannose and galactose 336:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 27 with O-glycosylated 291:304 arg1 types 315:319 six types 311:319 six types of linear 311:329 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 1 28 dep crassa 140:145 arg1 CBHI 168:171 CBHI 168:171 CBHI 168:171 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 1 28 dep crassa 140:145 arg1 I 165:165 cellobiohydrolase I 147:165 cellobiohydrolase I (CBHI) 147:172 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 2 29 theme O-glycans 361:369 arg1 types 342:346 three types 336:346 three types of branched, O-glycans containing approximately equal amounts of mannose and galactose 336:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 29 theme O-glycans 361:369 arg1 types 315:319 six types 311:319 six types of linear 311:329 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 30 theme galactose 425:433 arg1 amounts 402:408 approximately equal amounts 382:408 approximately equal amounts of mannose and galactose 382:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 30 theme galactose 425:433 arg1 galactose 425:433 galactose 425:433 galactose 425:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 30 theme galactose 425:433 arg1 mannose 413:419 mannose 413:419 mannose 413:419 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 3 31 theme protein-O-galactosyltransferase 584:614 arg1 existence 569:577 the existence 565:577 the existence of a protein-O-galactosyltransferase in N. crassa 565:627 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 32 theme equal 396:400 arg1 amounts 402:408 approximately equal amounts 382:408 approximately equal amounts of mannose and galactose 382:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 32 theme equal 396:400 arg1 galactose 425:433 galactose 425:433 galactose 425:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 32 theme equal 396:400 arg1 mannose 413:419 mannose 413:419 mannose 413:419 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 33 theme branched 351:358 arg1 O-glycans 361:369 branched, O-glycans 351:369 O-glycans 361:369 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 1 34 theme cellulose 272:280 arg1 presence 260:267 the presence 256:267 the presence of cellulose 256:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 35 theme end 536:538 arg1 galactoses 540:549 reducing end galactoses 527:549 reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 527:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 1 36 theme O-linked 105:112 arg1 glycans 114:120 the O-linked glycans 101:120 the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 101:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 0 37 theme Neurospora 46:55 arg1 crassa 57:62 Neurospora crassa 46:62 Neurospora crassa 46:62 O-Glycan analysis of cellobiohydrolase I from Neurospora crassa. 26762173 3 38 theme O-glycans 723:731 arg1 knowledge 701:709 the knowledge 697:709 the knowledge of the CBHI O-glycans 697:731 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 39 from evaluation 755:764 arg1 function 810:817 cellulase function 800:817 cellulase function 800:817 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 39 from evaluation 755:764 arg1 development 827:837 the development 823:837 the development of directed O-glycan/cellulase engineering 823:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 1 40 theme glycans 114:120 arg1 composition 86:96 the composition 82:96 the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 82:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 1 41 link O-linked 105:112 arg1 glycans 114:120 the O-linked glycans 101:120 the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 101:280 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 42 theme excellent 644:652 arg1 resources 662:670 the excellent genetic resources 640:670 the excellent genetic resources available for N. crassa 640:694 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 43 gly O-glycosylated 291:304 arg1 CBHI 283:286 CBHI 283:286 CBHI 283:286 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 3 44 theme cellulase 800:808 arg1 function 810:817 cellulase function 800:817 cellulase function 800:817 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 45 contain containing 371:380 arg2 galactose 425:433 galactose 425:433 galactose 425:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 45 contain containing 371:380 arg1 O-glycans 361:369 branched, O-glycans 351:369 O-glycans 361:369 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 45 contain containing 371:380 arg2 mannose 413:419 mannose 413:419 mannose 413:419 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 45 contain containing 371:380 arg2 amounts 402:408 approximately equal amounts 382:408 approximately equal amounts of mannose and galactose 382:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 1 46 theme Neurospora 129:138 arg1 crassa 140:145 the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 125:280 crassa 140:145 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 1 47 from composition 86:96 arg1 crassa 140:145 the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose 125:280 crassa 140:145 We describe here the composition of the O-linked glycans on the Neurospora crassa cellobiohydrolase I (CBHI), which accounts for approximately 40% of the protein secreted by cells growing in the presence of cellulose. 26762173 3 48 theme future 748:753 arg1 evaluation 755:764 the future evaluation 744:764 the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering 744:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 3 49 theme available 672:680 arg1 resources 662:670 the excellent genetic resources 640:670 the excellent genetic resources available for N. crassa 640:694 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 26762173 2 50 theme linear 324:329 arg1 types 342:346 three types 336:346 three types of branched, O-glycans containing approximately equal amounts of mannose and galactose 336:433 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 2 50 theme linear 324:329 arg1 types 315:319 six types 311:319 six types of linear 311:329 CBHI is O-glycosylated with six types of linear, and three types of branched, O-glycans containing approximately equal amounts of mannose and galactose. 26762173 3 51 theme directed 842:849 arg1 engineering 870:880 directed O-glycan/cellulase engineering 842:880 directed O-glycan/cellulase engineering 842:880 In addition to the classic fungal O-glycans with reducing end mannoses, we also identified reducing end galactoses which suggest the existence of a protein-O-galactosyltransferase in N. crassa Because of the excellent genetic resources available for N. crassa, the knowledge of the CBHI O-glycans may enable the future evaluation of the role of O-glycosylation on cellulase function and the development of directed O-glycan/cellulase engineering. 27663739 5 0 theme adjacent 784:791 arg1 or 781:782 or 781:782 or adjacent to Ser-rich domains (SRDs) 781:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 6 1 theme cell-signaling 940:953 arg1 proteins 955:962 cell-signaling proteins 940:962 cell-signaling proteins 940:962 Sixty-nine Ser-rich proteins were reproducibly enriched with AAL, including nucleoporins, mRNA-processing enzymes, and cell-signaling proteins. 27663739 8 2 theme Superresolution 1070:1084 arg1 microscopy 1086:1095 Superresolution microscopy 1070:1095 Superresolution microscopy 1070:1095 Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. 27663739 3 3 theme bradyzoites 523:533 arg1 nuclei 500:505 the nuclei 496:505 the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 496:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 0 4 theme pore 78:81 arg1 complexes 83:91 the nuclear pore complexes 66:91 the nuclear pore complexes of Toxoplasma gondii 66:112 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 6 5 theme Ser-rich 832:839 arg1 proteins 841:848 Sixty-nine Ser-rich proteins 821:848 Sixty-nine Ser-rich proteins 821:848 Sixty-nine Ser-rich proteins were reproducibly enriched with AAL, including nucleoporins, mRNA-processing enzymes, and cell-signaling proteins. 27663739 5 6 link O-linked 728:735 arg1 fucose 737:742 O-linked fucose 728:742 O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs) 728:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 0 7 theme Toxoplasma 96:105 arg1 complexes 83:91 the nuclear pore complexes 66:91 the nuclear pore complexes of Toxoplasma gondii 66:112 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 7 8 theme SRD-YFP 1012:1018 arg1 fusion 1020:1025 an SRD-YFP fusion 1009:1025 an SRD-YFP fusion 1009:1025 Two endogenous SRDs-containing proteins and an SRD-YFP fusion localize with AAL to the nuclear membrane. 27663739 2 9 from organization 359:370 arg1 nucleus 379:385 the nucleus 375:385 the nucleus 375:385 Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. 27663739 3 10 theme oocysts 560:566 arg1 nuclei 500:505 the nuclei 496:505 the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 496:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 5 11 theme Ser/Thr 753:759 arg1 residues 761:768 Ser/Thr residues 753:768 Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs) 753:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 5 12 theme AAL-enriched 682:693 arg1 fraction 695:702 the AAL-enriched fraction 678:702 the AAL-enriched fraction 678:702 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 9 13 from pathway 1277:1283 arg1 nucleocytosol 1292:1304 the nucleocytosol 1288:1304 the nucleocytosol of a eukaryote 1288:1319 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 1 14 theme Toxoplasma 115:124 arg1 parasite 153:160 an intracellular parasite 136:160 an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals 136:241 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 1 14 theme Toxoplasma 115:124 arg1 gondii 126:131 Toxoplasma gondii 115:131 Toxoplasma gondii 115:131 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 1 15 theme disseminated 174:185 arg1 infections 187:196 disseminated infections 174:196 disseminated infections 174:196 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 5 16 theme present 770:776 arg1 residues 761:768 Ser/Thr residues 753:768 Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs) 753:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 5 17 theme Ser-rich 796:803 arg1 SRDs 814:817 SRDs 814:817 SRDs 814:817 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 5 17 theme Ser-rich 796:803 arg1 domains 805:811 Ser-rich domains 796:811 Ser-rich domains (SRDs) 796:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 7 18 theme endogenous 969:978 arg1 proteins 996:1003 Two endogenous SRDs-containing proteins 965:1003 Two endogenous SRDs-containing proteins 965:1003 Two endogenous SRDs-containing proteins and an SRD-YFP fusion localize with AAL to the nuclear membrane. 27663739 0 19 theme O-fucosylated 0:12 arg1 glycoproteins 14:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 3 20 theme numerous 464:471 arg1 structures 482:491 numerous punctate structures 464:491 numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 464:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 3 21 theme fucose-binding 410:423 arg1 lectin 442:447 the fucose-binding Aleuria aurantia lectin 406:447 the fucose-binding Aleuria aurantia lectin (AAL) 406:453 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 3 21 theme fucose-binding 410:423 arg1 AAL 450:452 AAL 450:452 AAL 450:452 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 7 22 theme SRDs-containing 980:994 arg1 proteins 996:1003 Two endogenous SRDs-containing proteins 965:1003 Two endogenous SRDs-containing proteins 965:1003 Two endogenous SRDs-containing proteins and an SRD-YFP fusion localize with AAL to the nuclear membrane. 27663739 2 23 theme protein 351:357 arg1 organization 359:370 protein organization 351:370 protein organization in the nucleus 351:385 Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. 27663739 6 24 theme mRNA-processing 911:925 arg1 enzymes 927:933 mRNA-processing enzymes 911:933 mRNA-processing enzymes 911:933 Sixty-nine Ser-rich proteins were reproducibly enriched with AAL, including nucleoporins, mRNA-processing enzymes, and cell-signaling proteins. 27663739 3 25 theme Aleuria 425:431 arg1 lectin 442:447 the fucose-binding Aleuria aurantia lectin 406:447 the fucose-binding Aleuria aurantia lectin (AAL) 406:453 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 3 25 theme Aleuria 425:431 arg1 AAL 450:452 AAL 450:452 AAL 450:452 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 2 26 dep parasite 286:293 arg1 differentiation 295:309 differentiation 295:309 differentiation 295:309 Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. 27663739 8 27 theme AAL 1129:1131 arg1 signal 1133:1138 the AAL signal 1125:1138 the AAL signal 1125:1138 Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. 27663739 9 28 theme O-fucosylation 1262:1275 arg1 pathway 1277:1283 the O-fucosylation pathway 1258:1283 the O-fucosylation pathway in the nucleocytosol of a eukaryote 1258:1319 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 4 29 theme related 643:649 arg1 apicomplexans 651:663 more distantly related apicomplexans 628:663 more distantly related apicomplexans 628:663 AAL also binds to Hammondia and Neospora nuclei but not to more distantly related apicomplexans. 27663739 5 30 theme O-linked 728:735 arg1 fucose 737:742 O-linked fucose 728:742 O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs) 728:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 1 31 theme immunocompromised 213:229 arg1 individuals 231:241 immunocompromised individuals 213:241 immunocompromised individuals 213:241 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 0 32 theme close 47:51 arg1 proximity 53:61 close proximity 47:61 close proximity to the nuclear pore complexes of Toxoplasma gondii 47:112 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 5 33 from present 770:776 arg1 or 781:782 or 781:782 or adjacent to Ser-rich domains (SRDs) 781:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 0 34 gly O-fucosylated 0:12 arg1 glycoproteins 14:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 3 35 theme punctate 473:480 arg1 structures 482:491 numerous punctate structures 464:491 numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 464:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 8 36 theme signal 1133:1138 arg1 majority 1113:1120 the majority 1109:1120 the majority of the AAL signal 1109:1138 Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. 27663739 0 37 gly glycoproteins 14:26 arg1 glycoproteins 14:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins 0:26 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 7 38 theme nuclear 1052:1058 arg1 membrane 1060:1067 the nuclear membrane 1048:1067 the nuclear membrane 1048:1067 Two endogenous SRDs-containing proteins and an SRD-YFP fusion localize with AAL to the nuclear membrane. 27663739 3 39 theme aurantia 433:440 arg1 lectin 442:447 the fucose-binding Aleuria aurantia lectin 406:447 the fucose-binding Aleuria aurantia lectin (AAL) 406:453 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 3 39 theme aurantia 433:440 arg1 AAL 450:452 AAL 450:452 AAL 450:452 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 5 40 theme fraction 695:702 arg1 Analyses 666:673 Analyses 666:673 Analyses of the AAL-enriched fraction 666:702 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 4 41 dep Hammondia 587:595 arg1 nuclei 610:615 nuclei 610:615 nuclei 610:615 AAL also binds to Hammondia and Neospora nuclei but not to more distantly related apicomplexans. 27663739 2 42 theme gene 253:256 arg1 regulation 258:267 gene regulation 253:267 gene regulation 253:267 Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. 27663739 3 43 from structures 482:491 arg1 nuclei 500:505 the nuclei 496:505 the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 496:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 3 44 theme tachyzoites 510:520 arg1 nuclei 500:505 the nuclei 496:505 the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 496:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 27663739 8 45 theme nuclear 1166:1172 arg1 complexes 1179:1187 nuclear pore complexes 1166:1187 nuclear pore complexes 1166:1187 Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. 27663739 6 46 theme Sixty-nine 821:830 arg1 proteins 841:848 Sixty-nine Ser-rich proteins 821:848 Sixty-nine Ser-rich proteins 821:848 Sixty-nine Ser-rich proteins were reproducibly enriched with AAL, including nucleoporins, mRNA-processing enzymes, and cell-signaling proteins. 27663739 9 47 theme secreted 1208:1215 arg1 proteins 1217:1224 secreted proteins 1208:1224 secreted proteins with O-fucose 1208:1238 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 9 48 theme Host 1190:1193 arg1 cells 1195:1199 Host cells 1190:1199 Host cells 1190:1199 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 9 49 theme eukaryote 1311:1319 arg1 nucleocytosol 1292:1304 the nucleocytosol 1288:1304 the nucleocytosol of a eukaryote 1288:1319 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 9 50 with proteins 1217:1224 arg1 O-fucose 1231:1238 O-fucose 1231:1238 O-fucose 1231:1238 Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. 27663739 0 51 theme nuclear 70:76 arg1 complexes 83:91 the nuclear pore complexes 66:91 the nuclear pore complexes of Toxoplasma gondii 66:112 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 5 52 from or 781:782 arg1 present 770:776 present 770:776 present 770:776 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 10 53 theme gene 1417:1420 arg1 expression 1422:1431 gene expression 1417:1431 gene expression 1417:1431 Furthermore, these results suggest O-fucosylation is a mechanism by which proteins involved in gene expression accumulate near the NPC. 27663739 1 54 theme intracellular 139:151 arg1 parasite 153:160 an intracellular parasite 136:160 an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals 136:241 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 1 54 theme intracellular 139:151 arg1 gondii 126:131 Toxoplasma gondii 115:131 Toxoplasma gondii 115:131 Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. 27663739 8 55 theme pore 1174:1177 arg1 complexes 1179:1187 nuclear pore complexes 1166:1187 nuclear pore complexes 1166:1187 Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. 27663739 0 56 from assemblies 33:42 arg1 proximity 53:61 close proximity 47:61 close proximity to the nuclear pore complexes of Toxoplasma gondii 47:112 O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii. 27663739 5 57 attach present 770:776 arg1 or 781:782 or 781:782 or adjacent to Ser-rich domains (SRDs) 781:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 5 57 attach present 770:776 arg2 residues 761:768 Ser/Thr residues 753:768 Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs) 753:818 Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). 27663739 3 58 theme sporozoites 540:550 arg1 nuclei 500:505 the nuclei 496:505 the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts 496:566 Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. 29221760 3 0 gly fucosylated 414:424 arg1 species 426:432 the fucosylated species 410:432 the fucosylated species 410:432 Among the HMOs, the fucosylated species are the most abundant. 29221760 3 0 gly fucosylated 414:424 arg1 abundant 447:454 abundant 447:454 abundant 447:454 Among the HMOs, the fucosylated species are the most abundant. 29221760 8 1 theme % 1407:1407 arg1 substrate 1384:1392 the donor substrate 1374:1392 the donor substrate of 14% and 18%, respectively 1374:1421 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 8 2 dep produced 1293:1300 arg1 reaching 1343:1350 reaching 1343:1350 reaching molar yields based on the donor substrate of 14% and 18%, respectively 1343:1421 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 5 3 theme thermal 688:694 arg1 stability 696:704 thermal stability 688:704 thermal stability 688:704 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 6 4 theme human 876:880 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 4 theme human 876:880 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 7 5 gly transfucosylation 1142:1158 arg1 lactose 1163:1169 lactose 1163:1169 lactose using citrus xyloglucan as fucosyl donor 1163:1210 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 0 6 theme milk 111:114 arg1 oligosaccharides 116:131 human milk oligosaccharides 105:131 human milk oligosaccharides 105:131 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 6 7 theme milk 882:885 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 7 theme milk 882:885 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 5 8 theme stability 696:704 arg1 terms 653:657 terms 653:657 terms of substrate specificity and thermal stability 653:704 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 6 9 theme complex 868:874 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 9 theme complex 868:874 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 0 10 theme human 105:109 arg1 oligosaccharides 116:131 human milk oligosaccharides 105:131 human milk oligosaccharides 105:131 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 2 11 theme infant 296:301 arg1 health 303:308 infant health 296:308 infant health 296:308 HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. 29221760 6 12 from perfringens 808:818 arg1 CpAfc2 784:789 The α-l-1,3/4-fucosidase CpAfc2 759:789 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens 759:818 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 2 13 theme bovine 357:362 arg1 milk 364:367 bovine milk 357:367 bovine milk used for infant formula 357:391 HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. 29221760 8 14 theme donor 1378:1382 arg1 substrate 1384:1392 the donor substrate 1374:1392 the donor substrate of 14% and 18%, respectively 1374:1421 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 1 15 theme unique 182:187 arg1 family 189:194 a unique family 180:194 a unique family of bioactive lactose-based molecules present in human breast milk 180:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 0 16 theme oligosaccharides 116:131 arg1 production 91:100 enzymatic production 81:100 enzymatic production of human milk oligosaccharides 81:131 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 7 17 theme citrus 1177:1182 arg1 xyloglucan 1184:1193 citrus xyloglucan 1177:1193 citrus xyloglucan 1177:1193 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 5 18 theme hydrolase 600:608 arg1 family 610:615 glycosyl hydrolase family 29 591:618 glycosyl hydrolase family 29 591:618 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 8 19 theme Mfuc5 1270:1274 arg1 catalysis 1276:1284 Mfuc5 catalysis 1270:1284 Mfuc5 catalysis 1270:1284 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 7 20 theme lactose 1163:1169 arg1 transfucosylation 1142:1158 transfucosylation 1142:1158 transfucosylation of lactose using citrus xyloglucan as fucosyl donor 1142:1210 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 5 21 theme substrate 662:670 arg1 specificity 672:682 substrate specificity 662:682 substrate specificity 662:682 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 2 22 from milk 364:367 arg1 absent 345:350 absent 345:350 absent 345:350 HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. 29221760 6 23 with acceptor 1011:1018 arg1 yield 1031:1035 a 39% yield 1025:1035 a 39% yield 1025:1035 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 8 24 theme molar 1352:1356 arg1 yields 1358:1363 molar yields 1352:1363 molar yields based on the donor substrate of 14% and 18%, respectively 1352:1421 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 0 25 theme Substrate 0:8 arg1 specificity 10:20 Substrate specificity 0:20 Substrate specificity 0:20 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 5 26 theme specificity 672:682 arg1 terms 653:657 terms 653:657 terms of substrate specificity and thermal stability 653:704 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 1 27 theme bioactive 199:207 arg1 molecules 223:231 bioactive lactose-based molecules 199:231 bioactive lactose-based molecules present in human breast milk 199:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 0 28 theme transfucosylation 26:42 arg1 activity 44:51 transfucosylation activity 26:51 transfucosylation activity 26:51 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 7 29 theme fucosyl 1198:1204 arg1 donor 1206:1210 fucosyl donor 1198:1210 fucosyl donor 1198:1210 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 1 30 theme lactose-based 209:221 arg1 molecules 223:231 bioactive lactose-based molecules 199:231 bioactive lactose-based molecules present in human breast milk 199:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 6 31 theme % 1029:1029 arg1 yield 1031:1035 a 39% yield 1025:1035 a 39% yield 1025:1035 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 7 32 dep Fusarium 1066:1073 arg1 graminearum 1075:1085 graminearum 1075:1085 graminearum 1075:1085 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 1 33 theme molecules 223:231 arg1 family 189:194 a unique family 180:194 a unique family of bioactive lactose-based molecules present in human breast milk 180:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 8 34 theme unidentified 1305:1316 arg1 fucosyllactose 1327:1340 an unidentified, non-HMO fucosyllactose 1302:1340 an unidentified, non-HMO fucosyllactose 1302:1340 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 6 35 theme 39 1027:1028 arg1 % 1029:1029 % 1029:1029 % 1029:1029 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 8 36 dep unidentified 1305:1316 arg1 non-HMO 1319:1325 non-HMO 1319:1325 non-HMO 1319:1325 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 6 37 theme LNFP 938:941 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 37 theme LNFP 938:941 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 7 38 from Fusarium 1066:1073 arg1 FgFCO1 1054:1059 α-l-Fucosidases FgFCO1 1038:1059 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome 1038:1118 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 1 39 theme present 233:239 arg1 molecules 223:231 bioactive lactose-based molecules 199:231 bioactive lactose-based molecules present in human breast milk 199:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 3 40 theme fucosylated 414:424 arg1 species 426:432 the fucosylated species 410:432 the fucosylated species 410:432 Among the HMOs, the fucosylated species are the most abundant. 29221760 3 40 theme fucosylated 414:424 arg1 abundant 447:454 abundant 447:454 abundant 447:454 Among the HMOs, the fucosylated species are the most abundant. 29221760 4 41 theme biomimetic 547:556 arg1 HMOs 558:561 biomimetic HMOs 547:561 biomimetic HMOs 547:561 Transfucosylation catalysed by retaining α-l-fucosidases is a new route for manufacturing biomimetic HMOs. 29221760 7 42 from metagenome 1109:1118 arg1 FgFCO1 1054:1059 α-l-Fucosidases FgFCO1 1038:1059 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome 1038:1118 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 1 43 from present 233:239 arg1 milk 257:260 human breast milk 244:260 human breast milk 244:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 8 44 theme 2'-fucosyllactose 1243:1259 arg1 formation 1230:1238 formation 1230:1238 formation of 2'-fucosyllactose 1230:1259 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 29221760 1 45 from milk 257:260 arg1 present 233:239 present 233:239 present 233:239 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 1 46 theme Human 134:138 arg1 oligosaccharides 145:160 Human milk oligosaccharides 134:160 Human milk oligosaccharides (HMOs) 134:167 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 1 46 theme Human 134:138 arg1 HMOs 163:166 HMOs 163:166 HMOs 163:166 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 0 47 theme GH29 56:59 arg1 specificity 10:20 Substrate specificity 0:20 Substrate specificity 0:20 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 0 47 theme GH29 56:59 arg1 activity 44:51 transfucosylation activity 26:51 transfucosylation activity 26:51 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 7 48 from Mfuc5 1091:1095 arg1 FgFCO1 1054:1059 α-l-Fucosidases FgFCO1 1038:1059 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome 1038:1118 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 6 49 theme lacto-N-fucopentaose 913:932 arg1 formation 846:854 the formation 842:854 the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 842:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 50 theme Clostridium 796:806 arg1 perfringens 808:818 Clostridium perfringens 796:818 Clostridium perfringens 796:818 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 1 51 theme milk 140:143 arg1 oligosaccharides 145:160 Human milk oligosaccharides 134:160 Human milk oligosaccharides (HMOs) 134:167 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 1 51 theme milk 140:143 arg1 HMOs 163:166 HMOs 163:166 HMOs 163:166 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 6 52 theme structure 903:911 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 52 theme structure 903:911 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 2 53 theme major 275:279 arg1 importance 281:290 major importance 275:290 major importance 275:290 HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. 29221760 5 54 theme glycosyl 591:598 arg1 family 610:615 glycosyl hydrolase family 29 591:618 glycosyl hydrolase family 29 591:618 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 1 55 theme human 244:248 arg1 milk 257:260 human breast milk 244:260 human breast milk 244:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 7 56 theme soil 1104:1107 arg1 metagenome 1109:1118 a soil metagenome 1102:1118 a soil metagenome 1102:1118 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 6 57 theme oligosaccharide 887:901 arg1 lacto-N-fucopentaose 913:932 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II 859:935 the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) 859:945 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 6 57 theme oligosaccharide 887:901 arg1 II 943:944 LNFP II 938:944 LNFP II 938:944 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 5 58 from family 610:615 arg1 α-l-fucosidases 570:584 Seven α-l-fucosidases 564:584 Seven α-l-fucosidases from glycosyl hydrolase family 29 564:618 Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. 29221760 1 59 theme breast 250:255 arg1 milk 257:260 human breast milk 244:260 human breast milk 244:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 0 60 theme enzymatic 81:89 arg1 production 91:100 enzymatic production 81:100 enzymatic production of human milk oligosaccharides 81:131 Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides. 29221760 1 61 attach present 233:239 arg1 milk 257:260 human breast milk 244:260 human breast milk 244:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 1 61 attach present 233:239 arg2 molecules 223:231 bioactive lactose-based molecules 199:231 bioactive lactose-based molecules present in human breast milk 199:260 Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. 29221760 6 62 theme α-l-1,3/4-fucosidase 763:782 arg1 CpAfc2 784:789 The α-l-1,3/4-fucosidase CpAfc2 759:789 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens 759:818 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 2 63 theme infant 378:383 arg1 formula 385:391 infant formula 378:391 infant formula 378:391 HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. 29221760 6 64 theme fucosyl 973:979 arg1 donor 981:985 fucosyl donor 973:985 fucosyl donor 973:985 The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. 29221760 4 65 theme new 519:521 arg1 Transfucosylation 457:473 Transfucosylation 457:473 Transfucosylation catalysed by retaining α-l-fucosidases 457:512 Transfucosylation catalysed by retaining α-l-fucosidases is a new route for manufacturing biomimetic HMOs. 29221760 4 65 theme new 519:521 arg1 route 523:527 a new route 517:527 a new route for manufacturing biomimetic HMOs 517:561 Transfucosylation catalysed by retaining α-l-fucosidases is a new route for manufacturing biomimetic HMOs. 29221760 7 66 theme α-l-Fucosidases 1038:1052 arg1 FgFCO1 1054:1059 α-l-Fucosidases FgFCO1 1038:1059 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome 1038:1118 α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. 29221760 8 67 theme % 1399:1399 arg1 substrate 1384:1392 the donor substrate 1374:1392 the donor substrate of 14% and 18%, respectively 1374:1421 FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively. 28295836 7 0 theme isolated 1568:1575 arg1 coat 1587:1590 the isolated egg jelly coat 1564:1590 the isolated egg jelly coat 1564:1590 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 0 1 theme mass 189:192 arg1 spectrometry 194:205 tandem mass spectrometry 182:205 tandem mass spectrometry systems 182:213 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 2 2 theme present 359:365 arg1 study 367:371 the present study 355:371 the present study 355:371 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 4 3 theme electrospray 936:947 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 3 theme electrospray 936:947 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 1 4 theme carbohydrate-rich 252:268 arg1 layer 270:274 a carbohydrate-rich layer 250:274 a carbohydrate-rich layer 250:274 Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. 28295836 0 5 theme urchin 53:58 arg1 coat 37:40 the egg jelly coat 23:40 the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 23:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 5 6 theme permethylated 1159:1171 arg1 oligosaccharides 1173:1188 permethylated oligosaccharides 1159:1188 permethylated oligosaccharides on MALDI-TOF MS. Fragmentation 1159:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 0 7 theme tandem 182:187 arg1 spectrometry 194:205 tandem mass spectrometry 182:205 tandem mass spectrometry systems 182:213 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 4 8 theme capillary 904:912 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 8 theme capillary 904:912 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 9 theme oligosaccharides 1262:1277 arg1 characterization 1236:1251 structural characterization 1225:1251 structural characterization of these oligosaccharides 1225:1277 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 4 10 theme ESI-Ion 1001:1007 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 10 theme ESI-Ion 1001:1007 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 11 theme liquid 914:919 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 11 theme liquid 914:919 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 12 theme structural 1225:1234 arg1 characterization 1236:1251 structural characterization 1225:1251 structural characterization of these oligosaccharides 1225:1277 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 4 13 theme permethylation 767:780 arg1 reaction 782:789 permethylation reaction 767:789 permethylation reaction 767:789 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 14 from N-glycans 1022:1030 arg1 glycoproteins 1054:1066 the egg jelly coat glycoproteins 1035:1066 the egg jelly coat glycoproteins 1035:1066 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 7 15 theme mannose 1616:1622 arg1 type 1624:1627 the high mannose type 1607:1627 the high mannose type 1607:1627 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 2 16 theme extracellular 529:541 arg1 matrix 543:548 an alecithal-type extracellular matrix 511:548 an alecithal-type extracellular matrix 511:548 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 0 17 theme spectrometry 194:205 arg1 systems 207:213 tandem mass spectrometry systems 182:213 tandem mass spectrometry systems 182:213 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 1 18 gly glycoproteins 337:349 arg1 glycoproteins 337:349 glycoproteins 337:349 glycoproteins 337:349 Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. 28295836 7 19 theme jelly 1581:1585 arg1 coat 1587:1590 the isolated egg jelly coat 1564:1590 the isolated egg jelly coat 1564:1590 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 0 20 theme mass 95:98 arg1 spectrometry 100:111 MALDI-TOF mass spectrometry 85:111 MALDI-TOF mass spectrometry 85:111 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 6 21 theme MALDI-TOF-MS 1323:1334 arg1 spectra 1359:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 2 22 theme mass 390:393 arg1 strategies 409:418 two mass spectrometric strategies 386:418 two mass spectrometric strategies 386:418 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 0 23 theme MALDI-TOF 85:93 arg1 spectrometry 100:111 MALDI-TOF mass spectrometry 85:111 MALDI-TOF mass spectrometry 85:111 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 3 24 theme protein 711:717 arg1 chain 719:723 their protein chain 705:723 their protein chain 705:723 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 2 25 theme jelly 489:493 arg1 coat 495:498 the Paracentrotus lividus egg jelly coat 459:498 the Paracentrotus lividus egg jelly coat 459:498 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 6 26 theme software 1409:1416 arg1 suite 1418:1422 the GlycoWorkbench software suite 1390:1422 the GlycoWorkbench software suite 1390:1422 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 6 26 theme software 1409:1416 arg1 tool 1427:1430 a tool 1425:1430 a tool for building, displaying, and profiling glycan masses 1425:1484 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 5 27 theme ESI-Ion 1297:1303 arg1 MS/MS 1310:1314 ESI-Ion trap MS/MS 1297:1314 ESI-Ion trap MS/MS 1297:1314 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 5 28 from m/z 1108:1110 arg1 molecules 1095:1103 sodiated molecules 1086:1103 sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation 1086:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 2 29 theme alecithal-type 514:527 arg1 matrix 543:548 an alecithal-type extracellular matrix 511:548 an alecithal-type extracellular matrix 511:548 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 2 30 theme egg 485:487 arg1 coat 495:498 the Paracentrotus lividus egg jelly coat 459:498 the Paracentrotus lividus egg jelly coat 459:498 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 2 31 theme coat 495:498 arg1 N-glycosylation 440:454 the N-glycosylation 436:454 the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs 436:568 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 5 32 theme coat 1049:1052 arg1 glycoproteins 1054:1066 the egg jelly coat glycoproteins 1035:1066 the egg jelly coat glycoproteins 1035:1066 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 4 33 theme chromatography 921:934 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 33 theme chromatography 921:934 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 0 34 theme Profiling 0:8 arg1 N-glycans 10:18 Profiling N-glycans 0:18 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 0:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 4 35 theme MALDI-TOF 886:894 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 35 theme MALDI-TOF 886:894 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 7 36 theme coat 1587:1590 arg1 oligosaccharides 1544:1559 The oligosaccharides 1540:1559 The oligosaccharides of the isolated egg jelly coat 1540:1590 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 2 37 dep Paracentrotus 463:475 arg1 lividus 477:483 lividus 477:483 lividus 477:483 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 2 38 theme mammalian 555:563 arg1 eggs 565:568 mammalian eggs 555:568 mammalian eggs 555:568 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 1 39 theme jelly 288:292 arg1 coat 294:297 the jelly coat 284:297 the jelly coat 284:297 Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. 28295836 4 40 theme mass 976:979 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 40 theme mass 976:979 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 41 theme sodiated 1086:1093 arg1 molecules 1095:1103 sodiated molecules 1086:1103 sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation 1086:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 3 42 theme Egg 571:573 arg1 jelly 575:579 Egg jelly 571:579 Egg jelly 571:579 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 0 43 theme spectrometry 100:111 arg1 ionization-ion 162:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 3 44 theme F 656:656 arg1 treatment 669:677 peptide N-glycosidase F (PNGase-F) treatment 634:677 peptide N-glycosidase F (PNGase-F) treatment 634:677 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 5 45 from oligosaccharides 1173:1188 arg1 Fragmentation 1207:1219 MALDI-TOF MS. Fragmentation 1193:1219 MALDI-TOF MS. Fragmentation 1193:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 1 46 theme Sea 216:218 arg1 eggs 227:230 Sea urchin eggs 216:230 Sea urchin eggs 216:230 Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. 28295836 0 47 theme jelly 31:35 arg1 coat 37:40 the egg jelly coat 23:40 the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 23:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 0 48 theme liquid 127:132 arg1 chromatography 134:147 capillary liquid chromatography 117:147 capillary liquid chromatography 117:147 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 7 49 theme high 1611:1614 arg1 type 1624:1627 the high mannose type 1607:1627 the high mannose type 1607:1627 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 4 50 theme desorption/ionization 830:850 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 50 theme desorption/ionization 830:850 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 1 51 theme urchin 220:225 arg1 eggs 227:230 Sea urchin eggs 216:230 Sea urchin eggs 216:230 Sea urchin eggs are surrounded by a carbohydrate-rich layer, termed the jelly coat, that consists of polysaccharides and glycoproteins. 28295836 0 52 theme egg 27:29 arg1 coat 37:40 the egg jelly coat 23:40 the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 23:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 6 53 theme trap-MS/MS 1348:1357 arg1 spectra 1359:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 0 54 theme capillary 117:125 arg1 chromatography 134:147 capillary liquid chromatography 117:147 capillary liquid chromatography 117:147 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 3 55 theme peptide 634:640 arg1 F 656:656 peptide N-glycosidase F 634:656 peptide N-glycosidase F (PNGase-F) treatment 634:677 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 3 55 theme peptide 634:640 arg1 PNGase-F 659:666 PNGase-F 659:666 PNGase-F 659:666 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 4 56 theme laser 824:828 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 56 theme laser 824:828 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 2 57 theme spectrometric 395:407 arg1 strategies 409:418 two mass spectrometric strategies 386:418 two mass spectrometric strategies 386:418 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 5 58 gly glycoproteins 1054:1066 arg1 glycoproteins 1054:1066 the egg jelly coat glycoproteins 1035:1066 the egg jelly coat glycoproteins 1035:1066 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 5 59 theme MALDI-TOF 1193:1201 arg1 Fragmentation 1207:1219 MALDI-TOF MS. Fragmentation 1193:1219 MALDI-TOF MS. Fragmentation 1193:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 0 60 theme electrospray 149:160 arg1 ionization-ion 162:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 3 61 theme N-glycosidase 642:654 arg1 F 656:656 peptide N-glycosidase F 634:656 peptide N-glycosidase F (PNGase-F) treatment 634:677 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 3 61 theme N-glycosidase 642:654 arg1 PNGase-F 659:666 PNGase-F 659:666 PNGase-F 659:666 Egg jelly was isolated, lyophilized, and dialyzed, followed by peptide N-glycosidase F (PNGase-F) treatment to release N-glycans from their protein chain. 28295836 4 62 theme mass 867:870 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 62 theme mass 867:870 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 0 63 theme coat 37:40 arg1 N-glycans 10:18 Profiling N-glycans 0:18 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 0:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 5 64 theme MS. 1203:1205 arg1 Fragmentation 1207:1219 MALDI-TOF MS. Fragmentation 1193:1219 MALDI-TOF MS. Fragmentation 1193:1219 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 0 65 theme chromatography 134:147 arg1 ionization-ion 162:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion 85:175 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 2 66 theme Paracentrotus 463:475 arg1 coat 495:498 the Paracentrotus lividus egg jelly coat 459:498 the Paracentrotus lividus egg jelly coat 459:498 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 4 67 theme time-of-flight 852:865 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 67 theme time-of-flight 852:865 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 6 68 theme ESI-Ion 1340:1346 arg1 spectra 1359:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra 1323:1365 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 4 69 theme trap 964:967 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 69 theme trap 964:967 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 70 theme egg 1039:1041 arg1 glycoproteins 1054:1066 the egg jelly coat glycoproteins 1035:1066 the egg jelly coat glycoproteins 1035:1066 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 2 71 contain has 507:509 arg2 matrix 543:548 an alecithal-type extracellular matrix 511:548 an alecithal-type extracellular matrix 511:548 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 2 71 contain has 507:509 arg1 coat 495:498 the Paracentrotus lividus egg jelly coat 459:498 the Paracentrotus lividus egg jelly coat 459:498 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 6 72 theme GlycoWorkbench 1394:1407 arg1 suite 1418:1422 the GlycoWorkbench software suite 1390:1422 the GlycoWorkbench software suite 1390:1422 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 6 72 theme GlycoWorkbench 1394:1407 arg1 tool 1427:1430 a tool 1425:1430 a tool for building, displaying, and profiling glycan masses 1425:1484 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 4 73 theme ionization-ion 949:962 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 73 theme ionization-ion 949:962 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 7 74 theme egg 1577:1579 arg1 coat 1587:1590 the isolated egg jelly coat 1564:1590 the isolated egg jelly coat 1564:1590 The oligosaccharides of the isolated egg jelly coat were mainly of the high mannose type. 28295836 5 75 theme jelly 1043:1047 arg1 glycoproteins 1054:1066 the egg jelly coat glycoproteins 1035:1066 the egg jelly coat glycoproteins 1035:1066 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 4 76 theme tandem 969:974 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 76 theme tandem 969:974 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 0 77 theme sea 49:51 arg1 urchin 53:58 the sea urchin 45:58 the sea urchin Paracentrotus lividus 45:80 Profiling N-glycans of the egg jelly coat of the sea urchin Paracentrotus lividus by MALDI-TOF mass spectrometry and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectrometry systems. 28295836 6 78 theme oligosaccharide 1512:1526 arg1 structures 1528:1537 the original oligosaccharide structures 1499:1537 the original oligosaccharide structures 1499:1537 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 4 79 theme matrix-assisted 808:822 arg1 spectrometry 872:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 808:883 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 808:898 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 79 theme matrix-assisted 808:822 arg1 MS 896:897 MALDI-TOF MS 886:897 MALDI-TOF MS 886:897 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 5 80 theme trap 1305:1308 arg1 MS/MS 1310:1314 ESI-Ion trap MS/MS 1297:1314 ESI-Ion trap MS/MS 1297:1314 N-glycans in the egg jelly coat glycoproteins were indicated by sodiated molecules at m/z 1579.8, 1783.9, 1988.0, 2192.0, and 2397.1 for permethylated oligosaccharides on MALDI-TOF MS. Fragmentation and structural characterization of these oligosaccharides were performed by ESI-Ion trap MS/MS. 28295836 6 81 theme glycan 1472:1477 arg1 masses 1479:1484 glycan masses 1472:1484 glycan masses 1472:1484 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 2 82 gly N-glycosylation 440:454 arg1 coat 495:498 the Paracentrotus lividus egg jelly coat 459:498 the Paracentrotus lividus egg jelly coat 459:498 In the present study, we describe two mass spectrometric strategies to characterize the N-glycosylation of the Paracentrotus lividus egg jelly coat, which has an alecithal-type extracellular matrix like mammalian eggs. 28295836 6 83 theme original 1503:1510 arg1 structures 1528:1537 the original oligosaccharide structures 1499:1537 the original oligosaccharide structures 1499:1537 Then, MALDI-TOF-MS and ESI-Ion trap-MS/MS spectra were interpreted using the GlycoWorkbench software suite, a tool for building, displaying, and profiling glycan masses, to identify the original oligosaccharide structures. 28295836 4 84 theme CapLC 995:999 arg1 trap-MS/MS 1009:1018 CapLC ESI-Ion trap-MS/MS 995:1018 CapLC ESI-Ion trap-MS/MS 995:1018 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 28295836 4 84 theme CapLC 995:999 arg1 spectroscopy 981:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy 904:992 capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS) 904:1019 These N-glycans were then derivatized by permethylation reaction, and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and capillary liquid chromatography electrospray ionization-ion trap tandem mass spectroscopy (CapLC ESI-Ion trap-MS/MS). 27743366 2 0 theme " 335:335 arg1 N-glycans 337:345 "simple" N-glycans 328:345 "simple" N-glycans 328:345 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 7 1 theme further 1463:1469 arg1 insights 1471:1478 further insights 1463:1478 further insights 1463:1478 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 2 2 theme familiar 231:238 arg1 motifs 258:263 the more familiar vertebrate glycan motifs 222:263 the more familiar vertebrate glycan motifs 222:263 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 6 3 theme normal-phase 1274:1285 arg1 HPLC 1287:1290 normal-phase HPLC 1274:1290 normal-phase HPLC 1274:1290 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 7 4 theme treatments 1445:1454 arg1 use 1416:1418 The use 1412:1418 The use of enzymatic or chemical treatments 1412:1454 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 5 5 with extraction 934:943 arg1 resins 988:993 reversed phase resins 973:993 reversed phase resins 973:993 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 5 with extraction 934:943 arg1 carbon 962:967 graphitized carbon 950:967 graphitized carbon 950:967 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 6 theme simple 305:310 arg1 organisms 313:321 "simple" organisms 304:321 "simple" organisms 304:321 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 8 7 theme evolutionary 1694:1705 arg1 questions 1707:1715 evolutionary questions 1694:1715 evolutionary questions regarding the function of these glycans 1694:1755 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 6 8 theme single 1324:1329 arg1 structures 1352:1361 even single isomeric or isobaric structures 1319:1361 even single isomeric or isobaric structures 1319:1361 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 3 9 theme possible 533:540 arg1 range 542:546 the maximal possible range 521:546 the maximal possible range of structures 521:560 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 6 10 theme present 1245:1251 arg1 glycans 1237:1243 glycans 1237:1243 glycans present 1237:1251 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 5 11 theme liquid 1073:1078 arg1 HPLC 1096:1099 HPLC 1096:1099 HPLC 1096:1099 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 11 theme liquid 1073:1078 arg1 chromatography 1080:1093 high-pressure liquid chromatography 1059:1093 high-pressure liquid chromatography (HPLC) 1059:1100 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 12 theme vertebrate 240:249 arg1 motifs 258:263 the more familiar vertebrate glycan motifs 222:263 the more familiar vertebrate glycan motifs 222:263 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 5 13 theme solid-phase 922:932 arg1 extraction 934:943 Subsequent solid-phase extraction 911:943 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins 911:993 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 8 14 dep 100 1613:1615 arg1 to 1610:1611 to 1610:1611 to 1610:1611 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 4 15 attach present 814:820 arg2 α1,3-fucose 799:809 core α1,3-fucose 794:809 core α1,3-fucose 794:809 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 4 15 attach present 814:820 arg1 invertebrates 780:792 most invertebrates 775:792 most invertebrates 775:792 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 2 16 theme species 463:469 arg1 range 454:458 a range 452:458 a range of species 452:469 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 5 17 theme desorption/ionization 1128:1148 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 17 theme desorption/ionization 1128:1148 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 4 18 from release 826:832 arg1 glycopeptides 854:866 glycopeptides 854:866 glycopeptides with peptide 854:879 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 5 19 theme graphitized 950:960 arg1 carbon 962:967 graphitized carbon 950:967 graphitized carbon 950:967 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 20 theme oligosaccharides 363:378 arg1 structures 380:389 rather complex oligosaccharides structures 348:389 rather complex oligosaccharides structures 348:389 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 20 theme oligosaccharides 363:378 arg1 ones 427:430 zwitterionic and anionic ones 402:430 zwitterionic and anionic ones 402:430 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 5 21 theme Subsequent 911:920 arg1 extraction 934:943 Subsequent solid-phase extraction 911:943 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins 911:993 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 22 theme anionic 419:425 arg1 ones 427:430 zwitterionic and anionic ones 402:430 zwitterionic and anionic ones 402:430 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 0 23 theme Invertebrate 12:23 arg1 N-Glycans 37:45 Invertebrate and Protist N-Glycans 12:45 Invertebrate and Protist N-Glycans 12:45 Analysis of Invertebrate and Protist N-Glycans. 27743366 4 24 theme core 794:797 arg1 α1,3-fucose 799:809 core α1,3-fucose 794:809 core α1,3-fucose 794:809 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 2 25 theme zwitterionic 402:413 arg1 ones 427:430 zwitterionic and anionic ones 402:430 zwitterionic and anionic ones 402:430 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 0 26 theme Protist 29:35 arg1 N-Glycans 37:45 Invertebrate and Protist N-Glycans 12:45 Invertebrate and Protist N-Glycans 12:45 Analysis of Invertebrate and Protist N-Glycans. 27743366 5 27 theme mass 1165:1168 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 27 theme mass 1165:1168 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 28 theme antennal 171:178 arg1 modifications 180:192 Both core and antennal modifications 157:192 modifications 180:192 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 29 theme simple 329:334 arg1 N-glycans 337:345 "simple" N-glycans 328:345 "simple" N-glycans 328:345 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 6 30 theme MALDI-TOF 1388:1396 arg1 MS 1398:1399 MALDI-TOF MS 1388:1399 MALDI-TOF MS 1388:1399 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 5 31 theme matrix-assisted 1106:1120 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 31 theme matrix-assisted 1106:1120 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 32 theme N-glycans 1024:1032 arg1 classes 1013:1019 different classes 1003:1019 different classes of N-glycans 1003:1032 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 3 33 theme fractionation 658:670 arg1 steps 672:676 fractionation steps 658:676 fractionation steps 658:676 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 5 34 theme phase 982:986 arg1 resins 988:993 reversed phase resins 973:993 reversed phase resins 973:993 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 4 35 theme most 775:778 arg1 invertebrates 780:792 most invertebrates 775:792 most invertebrates 775:792 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 7 36 dep modifications 1515:1527 arg1 methylation 1541:1551 methylation 1541:1551 some glycan modifications (especially methylation) 1503:1552 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 4 37 located present 814:820 arg2 α1,3-fucose 799:809 core α1,3-fucose 794:809 core α1,3-fucose 794:809 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 4 37 located present 814:820 arg1 invertebrates 780:792 most invertebrates 775:792 most invertebrates 775:792 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 8 38 theme methods 1588:1594 arg1 battery 1577:1583 a battery 1575:1583 a battery of methods, sometimes up to 100 structures from a single organism 1575:1649 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 1 39 from protists 81:88 arg1 N-glycans 48:56 N-glycans 48:56 N-glycans from invertebrates and protists 48:88 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 6 40 dep reversed- 1261:1269 arg1 both 1296:1299 both 1296:1299 both 1296:1299 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 3 41 theme organisms 610:618 arg1 workflow 578:585 the analytical workflow 563:585 the analytical workflow for glycomics of these organisms 563:618 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 1 42 from invertebrates 63:75 arg1 N-glycans 48:56 N-glycans 48:56 N-glycans from invertebrates and protists 48:88 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 2 43 theme " 311:311 arg1 organisms 313:321 "simple" organisms 304:321 "simple" organisms 304:321 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 7 44 theme chemical 1436:1443 arg1 treatments 1445:1454 enzymatic or chemical treatments 1423:1454 enzymatic or chemical treatments 1423:1454 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 4 45 theme glycans 841:847 arg1 release 826:832 release 826:832 release of the glycans from glycopeptides with peptide:N-glycosidases A 826:896 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 2 46 located found 443:447 arg2 ones 427:430 zwitterionic and anionic ones 402:430 zwitterionic and anionic ones 402:430 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 46 located found 443:447 arg2 structures 380:389 rather complex oligosaccharides structures 348:389 rather complex oligosaccharides structures 348:389 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 46 located found 443:447 arg1 range 454:458 a range 452:458 a range of species 452:469 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 47 theme core 162:165 arg1 modifications 180:192 Both core and antennal modifications 157:192 modifications 180:192 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 3 48 theme optimized 496:504 arg1 elucidation 506:516 the optimized elucidation 492:516 the optimized elucidation of the maximal possible range of structures 492:560 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 7 49 theme enzymatic 1423:1431 arg1 treatments 1445:1454 enzymatic or chemical treatments 1423:1454 enzymatic or chemical treatments 1423:1454 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 3 50 theme range 542:546 arg1 elucidation 506:516 the optimized elucidation 492:516 the optimized elucidation of the maximal possible range of structures 492:560 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 6 51 theme isomeric 1331:1338 arg1 structures 1352:1361 even single isomeric or isobaric structures 1319:1361 even single isomeric or isobaric structures 1319:1361 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 3 52 theme maximal 525:531 arg1 range 542:546 the maximal possible range 521:546 the maximal possible range of structures 521:560 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 4 53 theme N-glycosidases 881:894 arg1 A 896:896 N-glycosidases A 881:896 release of the glycans from glycopeptides with peptide:N-glycosidases A 826:896 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 5 54 theme high-pressure 1059:1071 arg1 HPLC 1096:1099 HPLC 1096:1099 HPLC 1096:1099 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 54 theme high-pressure 1059:1071 arg1 chromatography 1080:1093 high-pressure liquid chromatography 1059:1093 high-pressure liquid chromatography (HPLC) 1059:1100 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 2 55 theme glycan 251:256 arg1 motifs 258:263 the more familiar vertebrate glycan motifs 222:263 the more familiar vertebrate glycan motifs 222:263 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 6 56 theme glycans 1237:1243 arg1 types 1216:1220 types 1216:1220 types 1216:1220 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 6 56 theme glycans 1237:1243 arg1 numbers 1226:1232 numbers 1226:1232 numbers 1226:1232 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 3 57 theme structures 551:560 arg1 range 542:546 the maximal possible range 521:546 the maximal possible range of structures 521:560 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 4 58 theme N-glycosidase 687:699 arg1 F 701:701 Peptide:N-glycosidase F 679:701 Peptide:N-glycosidase F 679:701 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 1 59 contain have 90:93 arg2 structures 109:118 unusual structures 101:118 unusual structures which present analytical challenges 101:154 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 1 59 contain have 90:93 arg1 N-glycans 48:56 N-glycans 48:56 N-glycans from invertebrates and protists 48:88 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 4 60 from invertebrates 780:792 arg1 present 814:820 present 814:820 present 814:820 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 1 61 theme analytical 134:143 arg1 challenges 145:154 analytical challenges 134:154 analytical challenges 134:154 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 6 62 dep types 1216:1220 arg1 the 1212:1214 the 1212:1214 the 1212:1214 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 4 63 from glycopeptides 854:866 arg1 release 826:832 release 826:832 release of the glycans from glycopeptides with peptide:N-glycosidases A 826:896 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 4 63 from glycopeptides 854:866 arg1 glycans 841:847 the glycans 837:847 the glycans from glycopeptides with peptide 837:879 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 8 64 theme glycans 1749:1755 arg1 function 1731:1738 the function 1727:1738 the function of these glycans 1727:1755 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 4 65 gly glycopeptides 854:866 arg2 glycopeptides 854:866 glycopeptides 854:866 glycopeptides with peptide 854:879 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 2 66 contain have 323:326 arg1 organisms 313:321 "simple" organisms 304:321 "simple" organisms 304:321 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 66 contain have 323:326 arg2 N-glycans 337:345 "simple" N-glycans 328:345 "simple" N-glycans 328:345 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 4 67 theme Peptide 679:685 arg1 F 701:701 Peptide:N-glycosidase F 679:701 Peptide:N-glycosidase F 679:701 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 5 68 theme time-of-flight 1150:1163 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 68 theme time-of-flight 1150:1163 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 6 69 theme isobaric 1343:1350 arg1 structures 1352:1361 even single isomeric or isobaric structures 1319:1361 even single isomeric or isobaric structures 1319:1361 Depending on the types and numbers of glycans present, either reversed- or normal-phase HPLC (or both in series) enable even single isomeric or isobaric structures to be separated prior to MALDI-TOF MS and MS/MS. 27743366 4 70 dep release 826:832 arg1 A 896:896 N-glycosidases A 881:896 release of the glycans from glycopeptides with peptide:N-glycosidases A 826:896 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 4 71 from present 814:820 arg1 invertebrates 780:792 most invertebrates 775:792 most invertebrates 775:792 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 2 72 theme complex 355:361 arg1 structures 380:389 rather complex oligosaccharides structures 348:389 rather complex oligosaccharides structures 348:389 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 2 72 theme complex 355:361 arg1 ones 427:430 zwitterionic and anionic ones 402:430 zwitterionic and anionic ones 402:430 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 1 73 theme unusual 101:107 arg1 structures 109:118 unusual structures 101:118 unusual structures which present analytical challenges 101:154 N-glycans from invertebrates and protists have often unusual structures which present analytical challenges. 27743366 2 74 from motifs 258:263 arg1 different 207:215 different 207:215 different 207:215 Both core and antennal modifications can be quite different from the more familiar vertebrate glycan motifs; thereby, contrary to the concept that "simple" organisms have "simple" N-glycans, rather complex oligosaccharides structures, including zwitterionic and anionic ones, have been found in a range of species. 27743366 5 75 theme laser 1122:1126 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 75 theme laser 1122:1126 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 8 76 from organism 1642:1649 arg1 methods 1588:1594 methods 1588:1594 methods 1588:1594 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 8 76 from organism 1642:1649 arg1 structures 1617:1626 up to 100 structures 1607:1626 up to 100 structures from a single organism 1607:1649 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 27743366 0 77 theme N-Glycans 37:45 arg1 Analysis 0:7 Analysis 0:7 Analysis of Invertebrate and Protist N-Glycans. 0:46 Analysis of Invertebrate and Protist N-Glycans. 27743366 5 78 theme different 1003:1011 arg1 classes 1013:1019 different classes 1003:1019 different classes of N-glycans 1003:1032 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 4 79 with glycopeptides 854:866 arg1 peptide 873:879 peptide 873:879 peptide 873:879 Peptide:N-glycosidase F is sufficient to isolate N-glycans from fungi and some protists, but in most invertebrates core α1,3-fucose is present, so release of the glycans from glycopeptides with peptide:N-glycosidases A is required. 27743366 7 80 theme glycan 1508:1513 arg1 modifications 1515:1527 some glycan modifications 1503:1527 some glycan modifications (especially methylation) 1503:1552 The use of enzymatic or chemical treatments allows further insights to be gained, although some glycan modifications (especially methylation) are resistant. 27743366 5 81 theme MALDI-TOF 1184:1192 arg1 spectrometry 1170:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 1106:1181 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) 1106:1196 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 81 theme MALDI-TOF 1184:1192 arg1 MS 1194:1195 MALDI-TOF MS 1184:1195 MALDI-TOF MS 1184:1195 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 5 82 theme reversed 973:980 arg1 resins 988:993 reversed phase resins 973:993 reversed phase resins 973:993 Subsequent solid-phase extraction with graphitized carbon and reversed phase resins enables different classes of N-glycans to be separated prior to high-pressure liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 27743366 3 83 theme analytical 567:576 arg1 workflow 578:585 the analytical workflow 563:585 the analytical workflow for glycomics of these organisms 563:618 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 3 84 theme sequential 635:644 arg1 release 646:652 sequential release 635:652 sequential release 635:652 Thus, to facilitate the optimized elucidation of the maximal possible range of structures, the analytical workflow for glycomics of these organisms should include sequential release and fractionation steps. 27743366 8 85 theme single 1635:1640 arg1 organism 1642:1649 a single organism 1633:1649 a single organism 1633:1649 Using a battery of methods, sometimes up to 100 structures from a single organism can be assigned, a complexity which raises evolutionary questions regarding the function of these glycans. 24794851 1 0 theme reduced 278:284 arg1 costs 286:290 reduced costs 278:290 reduced costs 278:290 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 9 1 theme deleted 1584:1590 arg1 residues 1613:1620 deleted plant-specific sugar residues 1584:1620 deleted plant-specific sugar residues 1584:1620 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 2 2 gly glycoproteins 313:325 arg1 glycoproteins 313:325 plant-made glycoproteins 302:325 plant-made glycoproteins 302:325 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 3 theme plant-made 302:311 arg1 glycoproteins 313:325 plant-made glycoproteins 302:325 plant-made glycoproteins 302:325 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 9 4 theme sugar 1607:1611 arg1 residues 1613:1620 deleted plant-specific sugar residues 1584:1620 deleted plant-specific sugar residues 1584:1620 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 6 5 theme transgenic 1057:1066 arg1 benthamiana 1071:1081 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 6 6 theme β-1,2-xylose 1026:1037 arg1 benthamiana 1071:1081 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 2 7 dep residues 368:375 arg1 α-1,3-fucose 400:411 α-1,3-fucose 400:411 α-1,3-fucose 400:411 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 7 dep residues 368:375 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 7 dep residues 368:375 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 10 8 theme N. 1723:1724 arg1 benthamiana 1726:1736 N. benthamiana 1723:1736 N. benthamiana 1723:1736 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 4 9 theme Nicotiana 853:861 arg1 plants 875:880 Nicotiana benthamiana plants 853:880 Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) 853:918 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 6 10 theme β-1,2-xylosyltransferase 1110:1133 arg1 co-suppression 1092:1105 co-suppression 1092:1105 co-suppression of β-1,2-xylosyltransferase 1092:1133 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 3 11 theme sugar 505:509 arg1 residues 511:518 these sugar residues 499:518 these sugar residues 499:518 Because it is likely that these sugar residues and glycan structures are immunogenic, many attempts have been made to delete them. 24794851 3 12 dep many 559:562 arg1 attempts 564:571 attempts 564:571 attempts 564:571 Because it is likely that these sugar residues and glycan structures are immunogenic, many attempts have been made to delete them. 24794851 9 13 theme ΔGMDΔXylT 1652:1660 arg1 plants 1662:1667 ΔGMDΔXylT plants 1652:1667 ΔGMDΔXylT plants 1652:1667 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 8 14 theme sugar 1348:1352 arg1 residues 1354:1361 deleted plant-specific sugar residues 1325:1361 deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1325:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 6 15 dep benthamiana 1071:1081 arg1 plant 1083:1087 plant 1083:1087 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 6 15 dep benthamiana 1071:1081 arg1 plant 1142:1146 ΔXylT plant 1136:1146 ΔXylT plant 1136:1146 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 8 16 from plants 1409:1414 arg1 protein 1386:1392 total soluble protein 1372:1392 total soluble protein from ΔGMDΔXylT plants 1372:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 4 17 dep repressing 741:750 arg1 Matsuo 921:926 Matsuo 921:926 Matsuo 921:926 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 17 dep repressing 741:750 arg1 Matsumura 932:940 Matsumura 932:940 Matsumura 932:940 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 17 dep repressing 741:750 arg1 Biotechnol 949:958 Plant Biotechnol 943:958 Plant Biotechnol 943:958 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 18 theme GDP-L-fucose 823:834 arg1 biosynthesis 836:847 GDP-L-fucose biosynthesis 823:847 GDP-L-fucose biosynthesis 823:847 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 1 19 gly glycoproteins 173:185 arg1 cytokines 223:231 cytokines 223:231 cytokines 223:231 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 1 19 gly glycoproteins 173:185 arg1 antibodies 208:217 therapeutic antibodies 196:217 therapeutic antibodies 196:217 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 1 19 gly glycoproteins 173:185 arg1 glycoproteins 173:185 pharmaceutical glycoproteins 158:185 pharmaceutical glycoproteins 158:185 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 8 20 theme total 1372:1376 arg1 protein 1386:1392 total soluble protein 1372:1392 total soluble protein from ΔGMDΔXylT plants 1372:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 4 21 theme plant-specific 661:674 arg1 residues 711:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues 657:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 657:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 10 22 theme plant-specific 1771:1784 arg1 residues 1792:1799 plant-specific sugar residues 1771:1799 plant-specific sugar residues 1771:1799 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 1 23 theme pharmaceutical 158:171 arg1 cytokines 223:231 cytokines 223:231 cytokines 223:231 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 1 23 theme pharmaceutical 158:171 arg1 antibodies 208:217 therapeutic antibodies 196:217 therapeutic antibodies 196:217 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 1 23 theme pharmaceutical 158:171 arg1 glycoproteins 173:185 pharmaceutical glycoproteins 158:185 pharmaceutical glycoproteins 158:185 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 8 24 located found 1363:1367 arg1 protein 1386:1392 total soluble protein 1372:1392 total soluble protein from ΔGMDΔXylT plants 1372:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 8 24 located found 1363:1367 arg2 residues 1354:1361 deleted plant-specific sugar residues 1325:1361 deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1325:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 7 25 theme sugar 1242:1246 arg1 residues 1248:1255 plant-specific sugar residues 1227:1255 plant-specific sugar residues 1227:1255 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 8 26 theme ΔGMDΔXylT 1399:1407 arg1 plants 1409:1414 ΔGMDΔXylT plants 1399:1414 ΔGMDΔXylT plants 1399:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 4 27 theme simultaneous 632:643 arg1 deletion 645:652 the simultaneous deletion 628:652 the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 628:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 9 28 theme Recombinant 1437:1447 arg1 mGM-CSF 1505:1511 mGM-CSF 1505:1511 mGM-CSF 1505:1511 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 28 theme Recombinant 1437:1447 arg1 factor 1497:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor 1437:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) 1437:1512 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 4 29 theme rGMD 883:886 arg1 plants 888:893 rGMD plants 883:893 rGMD plants 883:893 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 0 30 from Deletion 0:7 arg1 N-glycans 51:59 plant N-glycans 45:59 plant N-glycans 45:59 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 0 31 theme 4,6-dehydratase 92:106 arg1 repression 64:73 repression 64:73 repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes 64:141 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 9 32 theme granulocyte/macrophage-colony 1455:1483 arg1 mGM-CSF 1505:1511 mGM-CSF 1505:1511 mGM-CSF 1505:1511 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 32 theme granulocyte/macrophage-colony 1455:1483 arg1 factor 1497:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor 1437:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) 1437:1512 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 2 33 theme a 426:426 arg1 epitope 436:442 a Lewis a (Le(a)) epitope 418:442 a Lewis a (Le(a)) epitope 418:442 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 33 theme a 426:426 arg1 GlcNAc 465:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 0 34 theme β-1,2-xylosyltransferase 112:135 arg1 genes 137:141 β-1,2-xylosyltransferase genes 112:141 β-1,2-xylosyltransferase genes 112:141 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 7 35 theme plant-specific 1227:1240 arg1 residues 1248:1255 plant-specific sugar residues 1227:1255 plant-specific sugar residues 1227:1255 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 0 36 from residues 33:40 arg1 N-glycans 51:59 plant N-glycans 45:59 plant N-glycans 45:59 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 4 37 theme α-1,3-fucose 681:692 arg1 residues 711:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues 657:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 657:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 38 theme α-1,4-fucose 698:709 arg1 residues 711:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues 657:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 657:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 0 39 theme plant-specific 12:25 arg1 residues 33:40 plant-specific sugar residues 12:40 plant-specific sugar residues in plant N-glycans 12:59 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 0 40 theme residues 33:40 arg1 Deletion 0:7 Deletion 0:7 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 0:142 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 2 41 contain have 327:330 arg2 N-glycans 332:340 N-glycans 332:340 N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 332:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 41 contain have 327:330 arg1 glycoproteins 313:325 plant-made glycoproteins 302:325 plant-made glycoproteins 302:325 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 41 contain have 327:330 arg2 GlcNAc 465:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 41 contain have 327:330 arg2 epitope 436:442 a Lewis a (Le(a)) epitope 418:442 a Lewis a (Le(a)) epitope 418:442 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 9 42 theme monoclonal 1524:1533 arg1 hIgG 1553:1556 hIgG 1553:1556 hIgG 1553:1556 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 42 theme monoclonal 1524:1533 arg1 G 1550:1550 human monoclonal immunoglobulin G 1518:1550 human monoclonal immunoglobulin G (hIgG) 1518:1557 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 4 43 theme 4,6-dehydratase 770:784 arg1 gene 792:795 the GDP-D-mannose 4,6-dehydratase (GMD) gene 752:795 the GDP-D-mannose 4,6-dehydratase (GMD) gene 752:795 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 2 44 theme core 378:381 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 44 theme core 378:381 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 0 45 theme plant 45:49 arg1 N-glycans 51:59 plant N-glycans 45:59 plant N-glycans 45:59 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 1 46 theme many 248:251 arg1 advantages 253:262 many advantages 248:262 many advantages 248:262 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 6 47 theme present 991:997 arg1 study 999:1003 the present study 987:1003 the present study 987:1003 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 8 48 theme plant-specific 1333:1346 arg1 residues 1354:1361 deleted plant-specific sugar residues 1325:1361 deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1325:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 4 49 from deletion 645:652 arg1 epitopes 729:736 Le(a) epitopes 723:736 Le(a) epitopes 723:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 9 50 with N-glycans 1569:1577 arg1 residues 1613:1620 deleted plant-specific sugar residues 1584:1620 deleted plant-specific sugar residues 1584:1620 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 10 51 theme XylT 1709:1712 arg1 genes 1714:1718 the GMD and XylT genes 1697:1718 the GMD and XylT genes 1697:1718 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 9 52 theme plant-specific 1592:1605 arg1 residues 1613:1620 deleted plant-specific sugar residues 1584:1620 deleted plant-specific sugar residues 1584:1620 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 6 53 theme N. 1068:1069 arg1 benthamiana 1071:1081 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 5 54 dep J. 961:962 arg1 264-281 968:974 264-281 968:974 264-281 968:974 J., 9, 264-281, 2011). 24794851 5 54 dep J. 961:962 arg1 2011 977:980 2011 977:980 2011 977:980 J., 9, 264-281, 2011). 24794851 10 55 theme GMD 1701:1703 arg1 genes 1714:1718 the GMD and XylT genes 1697:1718 the GMD and XylT genes 1697:1718 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 1 56 theme therapeutic 196:206 arg1 antibodies 208:217 therapeutic antibodies 196:217 therapeutic antibodies 196:217 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 6 57 theme residue-repressed 1039:1055 arg1 benthamiana 1071:1081 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 2 58 theme plant-specific 347:360 arg1 α-1,3-fucose 400:411 α-1,3-fucose 400:411 α-1,3-fucose 400:411 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 58 theme plant-specific 347:360 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 58 theme plant-specific 347:360 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 6 59 theme core 1021:1024 arg1 benthamiana 1071:1081 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 8 60 with proportion 1296:1305 arg1 residues 1354:1361 deleted plant-specific sugar residues 1325:1361 deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1325:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 4 61 theme benthamiana 863:873 arg1 plants 875:880 Nicotiana benthamiana plants 853:880 Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) 853:918 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 6 62 theme ΔXylT 1136:1140 arg1 plant 1083:1087 plant 1083:1087 a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant) 1019:1147 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 6 62 theme ΔXylT 1136:1140 arg1 plant 1142:1146 ΔXylT plant 1136:1146 ΔXylT plant 1136:1146 In the present study, we generated a core β-1,2-xylose residue-repressed transgenic N. benthamiana plant by co-suppression of β-1,2-xylosyltransferase (ΔXylT plant). 24794851 3 63 theme glycan 524:529 arg1 structures 531:540 glycan structures 524:540 glycan structures 524:540 Because it is likely that these sugar residues and glycan structures are immunogenic, many attempts have been made to delete them. 24794851 8 64 theme soluble 1378:1384 arg1 protein 1386:1392 total soluble protein 1372:1392 total soluble protein from ΔGMDΔXylT plants 1372:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 7 65 theme ΔGMDΔXylT 1273:1281 arg1 plants 1283:1288 ΔGMDΔXylT plants 1273:1288 ΔGMDΔXylT plants 1273:1288 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 10 66 from repression 1683:1692 arg1 benthamiana 1726:1736 N. benthamiana 1723:1736 N. benthamiana 1723:1736 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 10 67 theme sugar 1786:1790 arg1 residues 1792:1799 plant-specific sugar residues 1771:1799 plant-specific sugar residues 1771:1799 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 2 68 with epitope 436:442 arg1 α-1,3-fucose 400:411 α-1,3-fucose 400:411 α-1,3-fucose 400:411 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 68 with epitope 436:442 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 68 with epitope 436:442 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 4 69 from epitopes 729:736 arg1 deletion 645:652 the simultaneous deletion 628:652 the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 628:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 1 70 theme glycoproteins 173:185 arg1 Production 144:153 Production 144:153 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants 144:242 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 0 71 theme GDP-D-mannose 78:90 arg1 4,6-dehydratase 92:106 GDP-D-mannose 4,6-dehydratase 78:106 GDP-D-mannose 4,6-dehydratase 78:106 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 2 72 with N-glycans 332:340 arg1 α-1,3-fucose 400:411 α-1,3-fucose 400:411 α-1,3-fucose 400:411 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 72 with N-glycans 332:340 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 72 with N-glycans 332:340 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 4 73 theme ΔGMD 907:910 arg1 plants 912:917 ΔGMD plants 907:917 ΔGMD plants 907:917 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 74 dep plants 875:880 arg1 plants 888:893 rGMD plants 883:893 rGMD plants 883:893 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 8 75 theme N-glycans 1310:1318 arg1 proportion 1296:1305 The proportion 1292:1305 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1292:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 24794851 9 76 theme mouse 1449:1453 arg1 mGM-CSF 1505:1511 mGM-CSF 1505:1511 mGM-CSF 1505:1511 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 76 theme mouse 1449:1453 arg1 factor 1497:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor 1437:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) 1437:1512 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 10 77 theme genes 1714:1718 arg1 repression 1683:1692 Simultaneous repression 1670:1692 Simultaneous repression of the GMD and XylT genes in N. benthamiana 1670:1736 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 0 78 from N-glycans 51:59 arg1 Deletion 0:7 Deletion 0:7 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 0:142 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 4 79 theme residues 711:718 arg1 deletion 645:652 the simultaneous deletion 628:652 the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 628:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 0 80 theme genes 137:141 arg1 repression 64:73 repression 64:73 repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes 64:141 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 4 81 from residues 711:718 arg1 epitopes 729:736 Le(a) epitopes 723:736 Le(a) epitopes 723:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 9 82 theme stimulating 1485:1495 arg1 mGM-CSF 1505:1511 mGM-CSF 1505:1511 mGM-CSF 1505:1511 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 82 theme stimulating 1485:1495 arg1 factor 1497:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor 1437:1502 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) 1437:1512 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 4 83 theme Le 723:724 arg1 epitopes 729:736 Le(a) epitopes 723:736 Le(a) epitopes 723:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 2 84 theme Lewis 420:424 arg1 a 426:426 Lewis a 420:426 a Lewis a (Le(a)) epitope 418:442 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 4 85 theme core 676:679 arg1 residues 711:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues 657:718 the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes 657:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 0 86 theme sugar 27:31 arg1 residues 33:40 plant-specific sugar residues 12:40 plant-specific sugar residues in plant N-glycans 12:59 Deletion of plant-specific sugar residues in plant N-glycans by repression of GDP-D-mannose 4,6-dehydratase and β-1,2-xylosyltransferase genes. 24794851 2 87 dep a 426:426 arg1 Le 429:430 Le 429:430 Le(a) 429:433 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 87 dep a 426:426 arg1 a 432:432 a 432:432 a 432:432 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 1 88 from Production 144:153 arg1 plants 237:242 plants 237:242 plants 237:242 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 4 89 theme Plant 943:947 arg1 Matsuo 921:926 Matsuo 921:926 Matsuo 921:926 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 89 theme Plant 943:947 arg1 Biotechnol 949:958 Plant Biotechnol 943:958 Plant Biotechnol 943:958 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 9 90 theme human 1518:1522 arg1 hIgG 1553:1556 hIgG 1553:1556 hIgG 1553:1556 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 90 theme human 1518:1522 arg1 G 1550:1550 human monoclonal immunoglobulin G 1518:1550 human monoclonal immunoglobulin G (hIgG) 1518:1557 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 91 theme immunoglobulin 1535:1548 arg1 hIgG 1553:1556 hIgG 1553:1556 hIgG 1553:1556 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 9 91 theme immunoglobulin 1535:1548 arg1 G 1550:1550 human monoclonal immunoglobulin G 1518:1550 human monoclonal immunoglobulin G (hIgG) 1518:1557 Recombinant mouse granulocyte/macrophage-colony stimulating factor (mGM-CSF) and human monoclonal immunoglobulin G (hIgG) harboring N-glycans with deleted plant-specific sugar residues were successfully produced in ΔGMDΔXylT plants. 24794851 7 92 theme ΔXylT 1171:1175 arg1 plants 1177:1182 ΔGMD and ΔXylT plants 1162:1182 ΔGMD and ΔXylT plants 1162:1182 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 2 93 theme Galβ 445:448 arg1 epitope 436:442 a Lewis a (Le(a)) epitope 418:442 a Lewis a (Le(a)) epitope 418:442 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 93 theme Galβ 445:448 arg1 GlcNAc 465:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 Galβ(1-3)[Fucα(1-4)]GlcNAc 445:470 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 1 94 contain has 244:246 arg1 Production 144:153 Production 144:153 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants 144:242 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 1 94 contain has 244:246 arg2 advantages 253:262 many advantages 248:262 many advantages 248:262 Production of pharmaceutical glycoproteins, such as therapeutic antibodies and cytokines, in plants has many advantages in safety and reduced costs. 24794851 4 95 theme GDP-D-mannose 756:768 arg1 GMD 787:789 GMD 787:789 GMD 787:789 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 4 95 theme GDP-D-mannose 756:768 arg1 4,6-dehydratase 770:784 GDP-D-mannose 4,6-dehydratase 756:784 the GDP-D-mannose 4,6-dehydratase (GMD) gene 752:795 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 7 96 theme ΔGMD 1162:1165 arg1 plants 1177:1182 ΔGMD and ΔXylT plants 1162:1182 ΔGMD and ΔXylT plants 1162:1182 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 4 97 theme a 726:726 arg1 epitopes 729:736 Le(a) epitopes 723:736 Le(a) epitopes 723:736 Previously, we reported the simultaneous deletion of the plant-specific core α-1,3-fucose and α-1,4-fucose residues in Le(a) epitopes by repressing the GDP-D-mannose 4,6-dehydratase (GMD) gene, which is associated with GDP-L-fucose biosynthesis, in Nicotiana benthamiana plants (rGMD plants, renamed to ΔGMD plants) (Matsuo and Matsumura, Plant Biotechnol. 24794851 2 98 theme sugar 362:366 arg1 α-1,3-fucose 400:411 α-1,3-fucose 400:411 α-1,3-fucose 400:411 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 98 theme sugar 362:366 arg1 residues 368:375 plant-specific sugar residues 347:375 plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) 347:412 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 2 98 theme sugar 362:366 arg1 β-1,2-xylose 383:394 core β-1,2-xylose 378:394 core β-1,2-xylose 378:394 However, plant-made glycoproteins have N-glycans with plant-specific sugar residues (core β-1,2-xylose and α-1,3-fucose) and a Lewis a (Le(a)) epitope, Galβ(1-3)[Fucα(1-4)]GlcNAc. 24794851 7 99 dep repressed 1262:1270 arg1 plants 1283:1288 ΔGMDΔXylT plants 1273:1288 ΔGMDΔXylT plants 1273:1288 By crossing ΔGMD and ΔXylT plants, we successfully generated plants in which plant-specific sugar residues were repressed (ΔGMDΔXylT plants). 24794851 10 100 theme Simultaneous 1670:1681 arg1 repression 1683:1692 Simultaneous repression 1670:1692 Simultaneous repression of the GMD and XylT genes in N. benthamiana 1670:1736 Simultaneous repression of the GMD and XylT genes in N. benthamiana is thus very useful for deleting plant-specific sugar residues. 24794851 8 101 theme deleted 1325:1331 arg1 residues 1354:1361 deleted plant-specific sugar residues 1325:1361 deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants 1325:1414 The proportion of N-glycans with deleted plant-specific sugar residues found in total soluble protein from ΔGMDΔXylT plants increased by 82.41%. 28314774 0 0 theme fibers 72:77 arg1 component 49:57 a major component 41:57 a major component of Mimivirus fibers 41:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 0 0 theme fibers 72:77 arg1 viosamine 28:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 2 1 theme functional 484:493 arg1 characterization 495:510 the functional characterization 480:510 the functional characterization of these enzymes 480:527 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 5 2 theme Sequence 938:945 arg1 analysis 947:954 Sequence analysis 938:954 Sequence analysis of the structural model of the L142 N-terminal domain 938:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 0 3 theme Mimivirus 62:70 arg1 fibers 72:77 Mimivirus fibers 62:77 Mimivirus fibers 62:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 6 4 theme moiety 1334:1339 arg1 transfer 1312:1319 the transfer 1308:1319 the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine 1308:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 5 theme mass 1186:1189 arg1 spectrometry 1191:1202 mass spectrometry 1186:1202 mass spectrometry 1186:1202 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 4 6 contain contains 844:851 arg2 glycosyltransferase 917:935 a predicted C-terminal glycosyltransferase 894:935 a predicted C-terminal glycosyltransferase 894:935 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 4 6 contain contains 844:851 arg2 domain 883:888 an N-terminal acyltransferase domain 853:888 an N-terminal acyltransferase domain 853:888 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 4 6 contain contains 844:851 arg1 protein 836:842 the L142 protein 827:842 the L142 protein 827:842 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 7 7 theme viosamine 1427:1435 arg1 presence 1404:1411 The presence 1400:1411 The presence of acetylated viosamine in vivo 1400:1443 The presence of acetylated viosamine in vivo has also been confirmed on the glycosylated viral fibers, using GC-MS and NMR. 28314774 4 8 theme C-terminal 906:915 arg1 glycosyltransferase 917:935 a predicted C-terminal glycosyltransferase 894:935 a predicted C-terminal glycosyltransferase 894:935 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 5 9 theme domain 1003:1008 arg1 model 974:978 the structural model 959:978 the structural model of the L142 N-terminal domain 959:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 5 10 theme N-terminal 992:1001 arg1 domain 1003:1008 the L142 N-terminal domain 983:1008 the L142 N-terminal domain 983:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 5 11 theme biosynthetic 1157:1168 arg1 pathways 1170:1177 biosynthetic pathways 1157:1177 biosynthetic pathways 1157:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 4 12 theme predicted 896:904 arg1 glycosyltransferase 917:935 a predicted C-terminal glycosyltransferase 894:935 a predicted C-terminal glycosyltransferase 894:935 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 6 13 theme L142 1244:1247 arg1 domain 1260:1265 the L142 N-terminal domain 1240:1265 the L142 N-terminal domain 1240:1265 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 13 theme L142 1244:1247 arg1 acetyltransferase 1278:1294 a sugar acetyltransferase 1270:1294 a sugar acetyltransferase 1270:1294 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 5 14 theme model 974:978 arg1 analysis 947:954 Sequence analysis 938:954 Sequence analysis of the structural model of the L142 N-terminal domain 938:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 2 15 theme enzymes 521:527 arg1 characterization 495:510 the functional characterization 480:510 the functional characterization of these enzymes 480:527 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 2 15 theme enzymes 521:527 arg1 identification 542:555 the full identification 533:555 the full identification of the glycans found in viral fibers 533:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 4 16 theme N-terminal 856:865 arg1 domain 883:888 an N-terminal acyltransferase domain 853:888 an N-terminal acyltransferase domain 853:888 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 5 17 theme significant 1020:1030 arg1 homology 1032:1039 significant homology 1020:1039 significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways 1020:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 3 18 located found 644:648 arg2 viosamine 621:629 viosamine 621:629 viosamine 621:629 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 3 18 located found 644:648 arg1 forms 662:666 acylated forms 653:666 acylated forms 653:666 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 8 19 theme sugar 1584:1588 arg1 acetyltransferase 1590:1606 a virally encoded sugar acetyltransferase 1566:1606 a virally encoded sugar acetyltransferase 1566:1606 This study represents the first report of a virally encoded sugar acetyltransferase. 28314774 7 20 theme viral 1489:1493 arg1 fibers 1495:1500 the glycosylated viral fibers 1472:1500 the glycosylated viral fibers 1472:1500 The presence of acetylated viosamine in vivo has also been confirmed on the glycosylated viral fibers, using GC-MS and NMR. 28314774 2 21 theme virus 383:387 arg1 genome 389:394 the virus genome 379:394 the virus genome 379:394 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 0 22 theme rare 4:7 arg1 component 49:57 a major component 41:57 a major component of Mimivirus fibers 41:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 0 22 theme rare 4:7 arg1 viosamine 28:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 8 23 theme encoded 1576:1582 arg1 acetyltransferase 1590:1606 a virally encoded sugar acetyltransferase 1566:1606 a virally encoded sugar acetyltransferase 1566:1606 This study represents the first report of a virally encoded sugar acetyltransferase. 28314774 2 24 located found 572:576 arg1 fibers 587:592 viral fibers 581:592 viral fibers 581:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 2 24 located found 572:576 arg2 glycans 564:570 the glycans 560:570 the glycans found in viral fibers 560:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 4 25 theme acyltransferase 867:881 arg1 domain 883:888 an N-terminal acyltransferase domain 853:888 an N-terminal acyltransferase domain 853:888 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 2 26 from cluster 368:374 arg1 genome 389:394 the virus genome 379:394 the virus genome 379:394 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 6 27 theme acetyl 1327:1332 arg1 moiety 1334:1339 an acetyl moiety 1324:1339 an acetyl moiety 1324:1339 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 0 28 theme N-acetylated 15:26 arg1 component 49:57 a major component 41:57 a major component of Mimivirus fibers 41:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 0 28 theme N-acetylated 15:26 arg1 viosamine 28:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 1 29 theme icosahedral 269:279 arg1 capsid 281:286 its icosahedral capsid 265:286 its icosahedral capsid 265:286 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 30 theme giant 84:88 arg1 system 142:147 an autonomous glycosylation system 114:147 an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 114:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 30 theme giant 84:88 arg1 Mimivirus 96:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 30 theme giant 84:88 arg1 responsible 171:181 responsible 171:181 responsible 171:181 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 8 31 theme first 1550:1554 arg1 report 1556:1561 the first report 1546:1561 the first report of a virally encoded sugar acetyltransferase 1546:1606 This study represents the first report of a virally encoded sugar acetyltransferase. 28314774 0 32 theme sugar 9:13 arg1 component 49:57 a major component 41:57 a major component of Mimivirus fibers 41:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 0 32 theme sugar 9:13 arg1 viosamine 28:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 6 33 theme N-terminal 1249:1258 arg1 domain 1260:1265 the L142 N-terminal domain 1240:1265 the L142 N-terminal domain 1240:1265 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 33 theme N-terminal 1249:1258 arg1 acetyltransferase 1278:1294 a sugar acetyltransferase 1270:1294 a sugar acetyltransferase 1270:1294 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 7 34 theme glycosylated 1476:1487 arg1 fibers 1495:1500 the glycosylated viral fibers 1472:1500 the glycosylated viral fibers 1472:1500 The presence of acetylated viosamine in vivo has also been confirmed on the glycosylated viral fibers, using GC-MS and NMR. 28314774 3 35 theme acylated 653:660 arg1 forms 662:666 acylated forms 653:666 acylated forms 653:666 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 1 36 theme virus 90:94 arg1 system 142:147 an autonomous glycosylation system 114:147 an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 114:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 36 theme virus 90:94 arg1 Mimivirus 96:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 36 theme virus 90:94 arg1 responsible 171:181 responsible 171:181 responsible 171:181 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 6 37 theme UDP-d-viosamine 1383:1397 arg1 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 37 theme UDP-d-viosamine 1383:1397 arg1 group 1374:1378 the C-4 amino group 1360:1378 the C-4 amino group of UDP-d-viosamine 1360:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 4 38 theme L142 831:834 arg1 protein 836:842 the L142 protein 827:842 the L142 protein 827:842 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 5 39 with homology 1032:1039 arg1 acetyltransferases 1071:1088 some characterized sugar acetyltransferases 1046:1088 some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways 1046:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 2 40 theme full 537:540 arg1 identification 542:555 the full identification 533:555 the full identification of the glycans found in viral fibers 533:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 2 41 theme nucleotide-sugar 426:441 arg1 production 443:452 nucleotide-sugar production 426:452 nucleotide-sugar production 426:452 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 7 42 gly glycosylated 1476:1487 arg1 fibers 1495:1500 the glycosylated viral fibers 1472:1500 the glycosylated viral fibers 1472:1500 The presence of acetylated viosamine in vivo has also been confirmed on the glycosylated viral fibers, using GC-MS and NMR. 28314774 0 43 theme major 43:47 arg1 component 49:57 a major component 41:57 a major component of Mimivirus fibers 41:77 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 0 43 theme major 43:47 arg1 viosamine 28:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine 0:36 The rare sugar N-acetylated viosamine is a major component of Mimivirus fibers. 28314774 2 44 theme viral 581:585 arg1 fibers 587:592 viral fibers 581:592 viral fibers 581:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 8 45 theme acetyltransferase 1590:1606 arg1 report 1556:1561 the first report 1546:1561 the first report of a virally encoded sugar acetyltransferase 1546:1606 This study represents the first report of a virally encoded sugar acetyltransferase. 28314774 2 46 theme glycan 458:463 arg1 formation 465:473 glycan formation 458:473 glycan formation 458:473 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 1 47 theme dideoxyhexose 303:315 arg1 viosamine 317:325 the dideoxyhexose viosamine 299:325 the dideoxyhexose viosamine 299:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 6 48 theme amino 1368:1372 arg1 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 48 theme amino 1368:1372 arg1 group 1374:1378 the C-4 amino group 1360:1378 the C-4 amino group of UDP-d-viosamine 1360:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 5 49 theme C-4 1106:1108 arg1 group 1116:1120 the C-4 amino group 1102:1120 the C-4 amino group 1102:1120 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 6 50 dep spectrometry 1191:1202 arg1 analyses 1212:1219 analyses 1212:1219 analyses 1212:1219 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 3 51 theme functional 765:774 arg1 annotations 776:786 our functional annotations 761:786 our functional annotations 761:786 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 6 52 theme C-4 1364:1366 arg1 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 UDP-d-viosamine 1383:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 52 theme C-4 1364:1366 arg1 group 1374:1378 the C-4 amino group 1360:1378 the C-4 amino group of UDP-d-viosamine 1360:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 5 53 theme amino 1110:1114 arg1 group 1116:1120 the C-4 amino group 1102:1120 the C-4 amino group 1102:1120 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 6 54 theme sugar 1272:1276 arg1 domain 1260:1265 the L142 N-terminal domain 1240:1265 the L142 N-terminal domain 1240:1265 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 6 54 theme sugar 1272:1276 arg1 acetyltransferase 1278:1294 a sugar acetyltransferase 1270:1294 a sugar acetyltransferase 1270:1294 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 2 55 theme Previous 328:335 arg1 studies 337:343 Previous studies 328:343 Previous studies 328:343 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 5 56 theme L142 987:990 arg1 domain 1003:1008 the L142 N-terminal domain 983:1008 the L142 N-terminal domain 983:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 1 57 theme complex 204:210 arg1 glycans 224:230 complex and unusual glycans 204:230 complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 204:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 2 58 theme glycans 564:570 arg1 characterization 495:510 the functional characterization 480:510 the functional characterization of these enzymes 480:527 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 2 58 theme glycans 564:570 arg1 identification 542:555 the full identification 533:555 the full identification of the glycans found in viral fibers 533:592 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 4 59 theme Bioinformatic 789:801 arg1 analyses 803:810 Bioinformatic analyses 789:810 Bioinformatic analyses 789:810 Bioinformatic analyses indicated that the L142 protein contains an N-terminal acyltransferase domain and a predicted C-terminal glycosyltransferase. 28314774 1 60 theme autonomous 117:126 arg1 system 142:147 an autonomous glycosylation system 114:147 an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 114:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 60 theme autonomous 117:126 arg1 Mimivirus 96:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 60 theme autonomous 117:126 arg1 responsible 171:181 responsible 171:181 responsible 171:181 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 5 61 theme structural 963:972 arg1 model 974:978 the structural model 959:978 the structural model of the L142 N-terminal domain 959:1008 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 5 62 theme characterized 1051:1063 arg1 acetyltransferases 1071:1088 some characterized sugar acetyltransferases 1046:1088 some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways 1046:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 1 63 theme glycosylation 128:140 arg1 system 142:147 an autonomous glycosylation system 114:147 an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 114:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 63 theme glycosylation 128:140 arg1 Mimivirus 96:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus 80:104 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 1 63 theme glycosylation 128:140 arg1 responsible 171:181 responsible 171:181 responsible 171:181 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 6 64 from acetyl-CoA 1346:1355 arg1 transfer 1312:1319 the transfer 1308:1319 the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine 1308:1397 Using mass spectrometry and NMR analyses, we confirmed that the L142 N-terminal domain is a sugar acetyltransferase, catalyzing the transfer of an acetyl moiety from acetyl-CoA to the C-4 amino group of UDP-d-viosamine. 28314774 1 65 theme unusual 216:222 arg1 glycans 224:230 complex and unusual glycans 204:230 complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 204:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 28314774 7 66 theme acetylated 1416:1425 arg1 viosamine 1427:1435 acetylated viosamine 1416:1435 acetylated viosamine 1416:1435 The presence of acetylated viosamine in vivo has also been confirmed on the glycosylated viral fibers, using GC-MS and NMR. 28314774 5 67 theme sugar 1065:1069 arg1 acetyltransferases 1071:1088 some characterized sugar acetyltransferases 1046:1088 some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways 1046:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 3 68 theme genes 698:702 arg1 one 687:689 one 687:689 one 687:689 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 3 68 theme genes 698:702 arg1 genes 698:702 the genes 694:702 the genes 694:702 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 3 68 theme genes 698:702 arg1 acyltransferase 720:734 an acyltransferase 717:734 an acyltransferase 717:734 Because viosamine is typically found in acylated forms, we suspected that one of the genes might encode an acyltransferase, providing directions to our functional annotations. 28314774 2 69 theme gene 363:366 arg1 cluster 368:374 a gene cluster 361:374 a gene cluster 361:374 Previous studies have identified a gene cluster in the virus genome, encoding enzymes involved in nucleotide-sugar production and glycan formation, but the functional characterization of these enzymes and the full identification of the glycans found in viral fibers remain incomplete. 28314774 5 70 dep bacillosamine 1129:1141 arg1 pathways 1170:1177 biosynthetic pathways 1157:1177 biosynthetic pathways 1157:1177 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 5 70 dep bacillosamine 1129:1141 arg1 the 1125:1127 the 1125:1127 the 1125:1127 Sequence analysis of the structural model of the L142 N-terminal domain indicated significant homology with some characterized sugar acetyltransferases that modify the C-4 amino group in the bacillosamine or perosamine biosynthetic pathways. 28314774 1 71 theme glycans 224:230 arg1 formation 191:199 the formation 187:199 the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine 187:325 The giant virus Mimivirus encodes an autonomous glycosylation system that is thought to be responsible for the formation of complex and unusual glycans composing the fibers surrounding its icosahedral capsid, including the dideoxyhexose viosamine. 27868361 2 0 theme partial 341:347 arg1 structures 349:358 the PGN partial structures 333:358 the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide 333:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 4 1 with octasaccharide 724:737 arg1 heptapeptide 755:766 the unique heptapeptide 744:766 the unique heptapeptide 744:766 Both the tetrasaccharide and octasaccharide with the unique heptapeptide were successfully synthesized for the first time. 27868361 1 2 theme host-recognition 240:255 arg1 proteins 257:264 proteins 257:264 proteins 257:264 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 0 3 theme Glycan 87:92 arg1 Elongation 94:103 Glycan Elongation 87:103 Glycan Elongation 87:103 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 2 4 theme PGN 337:339 arg1 structures 349:358 the PGN partial structures 333:358 the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide 333:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 5 theme chains 534:539 arg1 elongation 509:518 elongation 509:518 elongation of the glycan chains 509:539 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 4 6 theme first 806:810 arg1 time 812:815 the first time 802:815 the first time 802:815 Both the tetrasaccharide and octasaccharide with the unique heptapeptide were successfully synthesized for the first time. 27868361 2 7 theme glycan 527:532 arg1 chains 534:539 the glycan chains 523:539 the glycan chains 523:539 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 8 with tetrasaccharide 402:416 arg1 heptapeptide 454:465 a unique heptapeptide 445:465 a unique heptapeptide 445:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 4 9 theme unique 748:753 arg1 heptapeptide 755:766 the unique heptapeptide 744:766 the unique heptapeptide 744:766 Both the tetrasaccharide and octasaccharide with the unique heptapeptide were successfully synthesized for the first time. 27868361 0 10 from faecalis 55:62 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 0:104 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 4 11 dep tetrasaccharide 704:718 arg1 the 700:702 the 700:702 the 700:702 Both the tetrasaccharide and octasaccharide with the unique heptapeptide were successfully synthesized for the first time. 27868361 1 12 theme bacterial 170:178 arg1 wall 185:188 the bacterial cell wall 166:188 the bacterial cell wall conferring cell shape 166:210 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 4 13 with tetrasaccharide 704:718 arg1 heptapeptide 755:766 the unique heptapeptide 744:766 the unique heptapeptide 744:766 Both the tetrasaccharide and octasaccharide with the unique heptapeptide were successfully synthesized for the first time. 27868361 1 14 theme cell 180:183 arg1 wall 185:188 the bacterial cell wall 166:188 the bacterial cell wall conferring cell shape 166:210 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 2 15 theme unique 447:452 arg1 heptapeptide 454:465 a unique heptapeptide 445:465 a unique heptapeptide 445:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 1 16 theme wall 185:188 arg1 component 153:161 an essential structural component 129:161 an essential structural component 129:161 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 1 16 theme wall 185:188 arg1 Peptidoglycan 106:118 Peptidoglycan 106:118 Peptidoglycan (PGN) 106:124 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 0 17 theme Fragments 27:35 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 0:104 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 3 18 theme 4'-O-Fmoc-protected 552:570 arg1 disaccharide 572:583 a 4'-O-Fmoc-protected disaccharide 550:583 a 4'-O-Fmoc-protected disaccharide 550:583 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 18 theme 4'-O-Fmoc-protected 552:570 arg1 intermediate 609:620 the key intermediate 601:620 the key intermediate in this efficient synthetic pathway for preparing various PGN fragments 601:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 1 19 theme cell 201:204 arg1 shape 206:210 cell shape 201:210 cell shape 201:210 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 0 20 theme Peptidoglycan 13:25 arg1 Fragments 27:35 Peptidoglycan Fragments 13:35 Peptidoglycan Fragments 13:35 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 0 21 theme Enterococcus 42:53 arg1 faecalis 55:62 Enterococcus faecalis 42:62 Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation 42:103 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 1 22 theme bacterial 291:299 arg1 proteins 309:316 bacterial surface proteins 291:316 host-recognition proteins and receptors as well as bacterial surface proteins 240:316 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 3 23 theme various 672:678 arg1 fragments 684:692 various PGN fragments 672:692 various PGN fragments 672:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 24 theme key 605:607 arg1 disaccharide 572:583 a 4'-O-Fmoc-protected disaccharide 550:583 a 4'-O-Fmoc-protected disaccharide 550:583 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 24 theme key 605:607 arg1 intermediate 609:620 the key intermediate 601:620 the key intermediate in this efficient synthetic pathway for preparing various PGN fragments 601:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 25 from intermediate 609:620 arg1 pathway 650:656 this efficient synthetic pathway 625:656 this efficient synthetic pathway for preparing various PGN fragments 625:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 26 used utilized 589:596 arg2 disaccharide 572:583 a 4'-O-Fmoc-protected disaccharide 550:583 a 4'-O-Fmoc-protected disaccharide 550:583 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 26 used utilized 589:596 arg2 intermediate 609:620 the key intermediate 601:620 the key intermediate in this efficient synthetic pathway for preparing various PGN fragments 601:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 1 27 theme surface 301:307 arg1 proteins 309:316 bacterial surface proteins 291:316 host-recognition proteins and receptors as well as bacterial surface proteins 240:316 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 2 28 from faecalis 378:385 arg1 structures 349:358 the PGN partial structures 333:358 the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide 333:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 29 contain contain 392:398 arg2 octasaccharide 425:438 an octasaccharide 422:438 an octasaccharide with a unique heptapeptide 422:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 29 contain contain 392:398 arg2 tetrasaccharide 402:416 a tetrasaccharide 400:416 a tetrasaccharide 400:416 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 29 contain contain 392:398 arg1 faecalis 378:385 Enterococcus faecalis 365:385 Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide 365:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 2 30 with octasaccharide 425:438 arg1 heptapeptide 454:465 a unique heptapeptide 445:465 a unique heptapeptide 445:465 In this work, the PGN partial structures from Enterococcus faecalis that contain a tetrasaccharide and an octasaccharide with a unique heptapeptide were synthesized via an Fmoc-strategy for elongation of the glycan chains. 27868361 3 31 theme efficient 630:638 arg1 pathway 650:656 this efficient synthetic pathway 625:656 this efficient synthetic pathway for preparing various PGN fragments 625:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 32 theme PGN 680:682 arg1 fragments 684:692 various PGN fragments 672:692 various PGN fragments 672:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 3 33 theme synthetic 640:648 arg1 pathway 650:656 this efficient synthetic pathway 625:656 this efficient synthetic pathway for preparing various PGN fragments 625:692 Namely, a 4'-O-Fmoc-protected disaccharide was utilized as the key intermediate in this efficient synthetic pathway for preparing various PGN fragments. 27868361 1 34 theme essential 132:140 arg1 component 153:161 an essential structural component 129:161 an essential structural component 129:161 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 1 34 theme essential 132:140 arg1 Peptidoglycan 106:118 Peptidoglycan 106:118 Peptidoglycan (PGN) 106:124 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 0 35 with faecalis 55:62 arg1 Fmoc-Strategy 69:81 Fmoc-Strategy 69:81 Fmoc-Strategy 69:81 Synthesis of Peptidoglycan Fragments from Enterococcus faecalis with Fmoc-Strategy for Glycan Elongation. 27868361 1 36 theme structural 142:151 arg1 component 153:161 an essential structural component 129:161 an essential structural component 129:161 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 27868361 1 36 theme structural 142:151 arg1 Peptidoglycan 106:118 Peptidoglycan 106:118 Peptidoglycan (PGN) 106:124 Peptidoglycan (PGN) is an essential structural component of the bacterial cell wall conferring cell shape, which can be recognized by host-recognition proteins and receptors as well as bacterial surface proteins. 25437919 5 0 theme ovarian 802:808 arg1 tissue 810:815 normal ovarian tissue 795:815 normal ovarian tissue 795:815 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 7 1 theme cancer 1202:1207 arg1 subtypes 1182:1189 both serous and endometrioid histological subtypes 1140:1189 both serous and endometrioid histological subtypes of ovarian cancer 1140:1207 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 2 theme bisecting 1102:1110 arg1 glycosylation 1112:1124 bisecting glycosylation 1102:1124 bisecting glycosylation 1102:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 2 3 theme women 382:386 arg1 year 393:396 14 000 women each year 375:396 14 000 women each year 375:396 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 0 4 theme cancer 86:91 arg1 tissues 93:99 ovarian cancer tissues 78:99 ovarian cancer tissues 78:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 6 5 theme membrane 910:917 arg1 glycoproteins 919:931 the membrane glycoproteins 906:931 the membrane glycoproteins 906:931 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 0 6 theme novel 109:113 arg1 structures 115:124 novel structures 109:124 novel structures 109:124 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 4 7 theme bisecting 680:688 arg1 structures 706:715 bisecting N-linked glycan structures 680:715 bisecting N-linked glycan structures 680:715 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 5 8 theme cancer 766:771 arg1 tissues 773:779 ovarian cancer tissues 758:779 ovarian cancer tissues 758:779 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 6 9 theme glycan 944:949 arg1 structures 951:960 unique glycan structures 937:960 unique glycan structures 937:960 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 7 10 theme ovarian 1194:1200 arg1 cancer 1202:1207 ovarian cancer 1194:1207 ovarian cancer 1194:1207 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 6 11 theme capture 834:840 arg1 strategy 842:849 a lectin capture strategy 825:849 a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS 825:880 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 2 12 theme reproductive 313:324 arg1 malignancy 326:335 the deadliest reproductive malignancy 299:335 the deadliest reproductive malignancy among women in the U.S. 299:359 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 2 12 theme reproductive 313:324 arg1 cancer 289:294 Ovarian cancer 281:294 Ovarian cancer 281:294 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 7 13 with glycoproteins 1083:1095 arg1 glycosylation 1112:1124 bisecting glycosylation 1102:1124 bisecting glycosylation 1102:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 14 theme signaling 1259:1267 arg1 Wnt 1293:1295 Wnt 1293:1295 Wnt 1293:1295 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 14 theme signaling 1259:1267 arg1 pathways 1269:1276 signaling pathways 1259:1276 signaling pathways such as Notch, Wnt, and TGFβ 1259:1305 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 14 theme signaling 1259:1267 arg1 Notch 1286:1290 Notch 1286:1290 Notch 1286:1290 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 14 theme signaling 1259:1267 arg1 TGFβ 1302:1305 TGFβ 1302:1305 TGFβ 1302:1305 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 6 15 theme unique 937:942 arg1 structures 951:960 unique glycan structures 937:960 unique glycan structures 937:960 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 7 16 theme histological 1169:1180 arg1 subtypes 1182:1189 both serous and endometrioid histological subtypes 1140:1189 both serous and endometrioid histological subtypes of ovarian cancer 1140:1207 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 6 17 theme lectin 827:832 arg1 strategy 842:849 a lectin capture strategy 825:849 a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS 825:880 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 2 18 theme deadliest 303:311 arg1 malignancy 326:335 the deadliest reproductive malignancy 299:335 the deadliest reproductive malignancy among women in the U.S. 299:359 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 2 18 theme deadliest 303:311 arg1 cancer 289:294 Ovarian cancer 281:294 Ovarian cancer 281:294 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 3 19 theme high 440:443 arg1 rates 455:459 high mortality rates 440:459 high mortality rates 440:459 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 1 20 theme epithelial 187:196 arg1 cells 213:217 epithelial ovarian cancer cells 187:217 epithelial ovarian cancer cells 187:217 Biomarkers capable of detecting and targeting epithelial ovarian cancer cells for diagnostics and therapeutics would be extremely valuable. 25437919 3 21 theme presenting 416:425 arg1 symptoms 427:434 presenting symptoms 416:434 presenting symptoms 416:434 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 1 22 theme ovarian 198:204 arg1 cells 213:217 epithelial ovarian cancer cells 187:217 epithelial ovarian cancer cells 187:217 Biomarkers capable of detecting and targeting epithelial ovarian cancer cells for diagnostics and therapeutics would be extremely valuable. 25437919 6 23 theme cancer 1017:1022 arg1 tissues 1024:1030 human ovarian cancer tissues 1003:1030 human ovarian cancer tissues 1003:1030 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 1 24 theme cancer 206:211 arg1 cells 213:217 epithelial ovarian cancer cells 187:217 epithelial ovarian cancer cells 187:217 Biomarkers capable of detecting and targeting epithelial ovarian cancer cells for diagnostics and therapeutics would be extremely valuable. 25437919 0 25 theme Glycomic 0:7 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues 0:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 7 26 theme endometrioid 1156:1167 arg1 subtypes 1182:1189 both serous and endometrioid histological subtypes 1140:1189 both serous and endometrioid histological subtypes of ovarian cancer 1140:1207 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 3 27 theme mortality 445:453 arg1 rates 455:459 high mortality rates 440:459 high mortality rates 440:459 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 0 28 theme membrane 21:28 arg1 glycoproteins 30:42 membrane glycoproteins 21:42 membrane glycoproteins 21:42 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 7 29 gly glycoproteins 1083:1095 arg1 glycoproteins 1083:1095 membrane glycoproteins 1074:1095 membrane glycoproteins with bisecting glycosylation 1074:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 4 30 theme Mgat3 597:601 arg1 gene 603:606 the Mgat3 gene 593:606 the Mgat3 gene 593:606 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 7 31 theme serous 1145:1150 arg1 subtypes 1182:1189 both serous and endometrioid histological subtypes 1140:1189 both serous and endometrioid histological subtypes of ovarian cancer 1140:1207 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 0 32 from tissues 93:99 arg1 glycosylation 59:71 bisecting glycosylation 49:71 bisecting glycosylation from ovarian cancer tissues 49:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 3 33 theme earlier 485:491 arg1 diagnosis 493:501 earlier diagnosis 485:501 earlier diagnosis 485:501 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 3 34 theme symptoms 427:434 arg1 rates 455:459 high mortality rates 440:459 high mortality rates 440:459 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 3 34 theme symptoms 427:434 arg1 lack 408:411 the lack 404:411 the lack of presenting symptoms 404:434 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 4 35 theme glycan 699:704 arg1 structures 706:715 bisecting N-linked glycan structures 680:715 bisecting N-linked glycan structures 680:715 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 4 36 theme N-linked 690:697 arg1 structures 706:715 bisecting N-linked glycan structures 680:715 bisecting N-linked glycan structures 680:715 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 0 37 theme glycoproteins 30:42 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues 0:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 4 38 theme glycosyltransferase 547:565 arg1 responsible 611:621 responsible 611:621 responsible 611:621 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 4 38 theme glycosyltransferase 547:565 arg1 GnT-III 574:580 The glycosyltransferase enzyme GnT-III 543:580 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene 543:606 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 4 39 link N-linked 690:697 arg1 structures 706:715 bisecting N-linked glycan structures 680:715 bisecting N-linked glycan structures 680:715 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 7 40 theme membrane 1074:1081 arg1 glycoproteins 1083:1095 membrane glycoproteins 1074:1095 membrane glycoproteins with bisecting glycosylation 1074:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 5 41 theme ovarian 758:764 arg1 tissues 773:779 ovarian cancer tissues 758:779 ovarian cancer tissues 758:779 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 7 42 theme glycoproteins 1083:1095 arg1 common 1130:1135 common 1130:1135 common 1130:1135 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 7 42 theme glycoproteins 1083:1095 arg1 majority 1062:1069 the majority 1058:1069 the majority of membrane glycoproteins with bisecting glycosylation 1058:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 3 43 theme improved 507:514 arg1 treatment 516:524 improved treatment 507:524 improved treatment of this disease 507:540 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 0 44 theme bisecting 49:57 arg1 glycosylation 59:71 bisecting glycosylation 49:71 bisecting glycosylation from ovarian cancer tissues 49:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 6 45 from amplification 986:998 arg1 tissues 1024:1030 human ovarian cancer tissues 1003:1030 human ovarian cancer tissues 1003:1030 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 0 46 with analysis 9:16 arg1 glycosylation 59:71 bisecting glycosylation 49:71 bisecting glycosylation from ovarian cancer tissues 49:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 7 47 gly glycosylation 1112:1124 arg1 glycoproteins 1083:1095 membrane glycoproteins 1074:1095 membrane glycoproteins with bisecting glycosylation 1074:1124 Our data illustrate that the majority of membrane glycoproteins with bisecting glycosylation are common to both serous and endometrioid histological subtypes of ovarian cancer, and several have been reported to participate in signaling pathways such as Notch, Wnt, and TGFβ. 25437919 6 48 theme ovarian 1009:1015 arg1 tissues 1024:1030 human ovarian cancer tissues 1003:1030 human ovarian cancer tissues 1003:1030 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 4 49 theme enzyme 567:572 arg1 responsible 611:621 responsible 611:621 responsible 611:621 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 4 49 theme enzyme 567:572 arg1 GnT-III 574:580 The glycosyltransferase enzyme GnT-III 543:580 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene 543:606 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 5 50 theme GnT-III 718:724 arg1 expression 731:740 GnT-III mRNA expression 718:740 GnT-III mRNA expression 718:740 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 0 51 gly glycoproteins 30:42 arg1 glycoproteins 30:42 membrane glycoproteins 21:42 membrane glycoproteins 21:42 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 5 52 theme mRNA 726:729 arg1 expression 731:740 GnT-III mRNA expression 718:740 GnT-III mRNA expression 718:740 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 1 53 theme capable 152:158 arg1 Biomarkers 141:150 Biomarkers 141:150 Biomarkers capable of detecting and targeting epithelial ovarian cancer cells for diagnostics and therapeutics 141:250 Biomarkers capable of detecting and targeting epithelial ovarian cancer cells for diagnostics and therapeutics would be extremely valuable. 25437919 2 54 from malignancy 326:335 arg1 U.S. 356:359 the U.S. 352:359 the U.S. 352:359 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 6 55 gly glycoproteins 919:931 arg1 glycoproteins 919:931 the membrane glycoproteins 906:931 the membrane glycoproteins 906:931 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 0 56 theme ovarian 78:84 arg1 tissues 93:99 ovarian cancer tissues 78:99 ovarian cancer tissues 78:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 2 57 theme Ovarian 281:287 arg1 malignancy 326:335 the deadliest reproductive malignancy 299:335 the deadliest reproductive malignancy among women in the U.S. 299:359 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 2 57 theme Ovarian 281:287 arg1 cancer 289:294 Ovarian cancer 281:294 Ovarian cancer 281:294 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 3 58 theme disease 534:540 arg1 diagnosis 493:501 earlier diagnosis 485:501 earlier diagnosis 485:501 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 3 58 theme disease 534:540 arg1 treatment 516:524 improved treatment 507:524 improved treatment of this disease 507:540 Both the lack of presenting symptoms and high mortality rates illustrate the need for earlier diagnosis and improved treatment of this disease. 25437919 4 59 theme GlcNAc 643:648 arg1 addition 631:638 the addition 627:638 the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures 627:715 The glycosyltransferase enzyme GnT-III encoded by the Mgat3 gene is responsible for the addition of GlcNAc (N-acetylglucosamine) to form bisecting N-linked glycan structures. 25437919 5 60 theme normal 795:800 arg1 tissue 810:815 normal ovarian tissue 795:815 normal ovarian tissue 795:815 GnT-III mRNA expression is amplified in ovarian cancer tissues compared with normal ovarian tissue. 25437919 6 61 theme GnT-III 978:984 arg1 amplification 986:998 GnT-III amplification 978:998 GnT-III amplification in human ovarian cancer tissues 978:1030 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25437919 0 62 gly glycosylation 59:71 arg1 analysis 9:16 Glycomic analysis 0:16 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues 0:99 Glycomic analysis of membrane glycoproteins with bisecting glycosylation from ovarian cancer tissues reveals novel structures and functions. 25437919 2 63 theme each 388:391 arg1 year 393:396 14 000 women each year 375:396 14 000 women each year 375:396 Ovarian cancer is the deadliest reproductive malignancy among women in the U.S., killing over 14 000 women each year. 25437919 6 64 theme human 1003:1007 arg1 tissues 1024:1030 human ovarian cancer tissues 1003:1030 human ovarian cancer tissues 1003:1030 We use a lectin capture strategy coupled to nano-ESI-RPLC-MS/MS to isolate and identify the membrane glycoproteins and unique glycan structures associated with GnT-III amplification in human ovarian cancer tissues. 25210812 4 0 theme residue 483:489 arg1 A 491:491 residue A 483:491 residue A 483:491 Precursors to residue A were obtained from L-glucose. 25210812 3 1 from strategy 421:428 arg1 combination 433:443 combination 433:443 combination with imidate chemistry 433:466 All targets were synthesized using a postglycosylation oxidation strategy in combination with imidate chemistry. 25210812 0 2 theme Shigella 67:74 arg1 sonnei 76:81 Shigella sonnei 67:81 Shigella sonnei 67:81 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 6 3 theme one-step 590:597 arg1 deprotection 618:629 A one-step Pd(OH)2/C-mediated deprotection 588:629 A one-step Pd(OH)2/C-mediated deprotection 588:629 A one-step Pd(OH)2/C-mediated deprotection provided the propyl glycoside targets. 25210812 0 4 theme chain 87:91 arg1 elongation 93:102 chain elongation 87:102 chain elongation 87:102 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 3 5 theme imidate 450:456 arg1 chemistry 458:466 imidate chemistry 450:466 imidate chemistry 450:466 All targets were synthesized using a postglycosylation oxidation strategy in combination with imidate chemistry. 25210812 1 6 theme zwitterionic 155:166 arg1 repeat 181:186 a zwitterionic disaccharide repeat 153:186 a zwitterionic disaccharide repeat encompassing two rare monosaccharides 153:224 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 2 7 theme trisaccharides 265:278 arg1 synthesis 231:239 The synthesis 227:239 The synthesis 227:239 The synthesis of the AB repeat and of trisaccharides ABA' and B'AB, which validates chain elongation at either end, is reported. 25210812 5 8 dep AAT 527:529 arg1 acceptor 545:552 acceptor 545:552 acceptor 545:552 The AAT (B) donor and acceptor were obtained from D-glucosamine. 25210812 5 8 dep AAT 527:529 arg1 donor 535:539 donor 535:539 donor 535:539 The AAT (B) donor and acceptor were obtained from D-glucosamine. 25210812 5 8 dep AAT 527:529 arg1 B 532:532 B 532:532 B 532:532 The AAT (B) donor and acceptor were obtained from D-glucosamine. 25210812 1 9 theme disaccharide 168:179 arg1 repeat 181:186 a zwitterionic disaccharide repeat 153:186 a zwitterionic disaccharide repeat encompassing two rare monosaccharides 153:224 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 6 10 theme glycoside 651:659 arg1 targets 661:667 the propyl glycoside targets 640:667 the propyl glycoside targets 640:667 A one-step Pd(OH)2/C-mediated deprotection provided the propyl glycoside targets. 25210812 3 11 theme postglycosylation 393:409 arg1 strategy 421:428 a postglycosylation oxidation strategy 391:428 a postglycosylation oxidation strategy in combination with imidate chemistry 391:466 All targets were synthesized using a postglycosylation oxidation strategy in combination with imidate chemistry. 25210812 0 12 theme zwitterionic 17:28 arg1 unit 40:43 the zwitterionic repeating unit 13:43 the zwitterionic repeating unit of the O-antigen 13:60 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 6 13 theme propyl 644:649 arg1 targets 661:667 the propyl glycoside targets 640:667 the propyl glycoside targets 640:667 A one-step Pd(OH)2/C-mediated deprotection provided the propyl glycoside targets. 25210812 3 14 with combination 433:443 arg1 chemistry 458:466 imidate chemistry 450:466 imidate chemistry 450:466 All targets were synthesized using a postglycosylation oxidation strategy in combination with imidate chemistry. 25210812 0 15 from sonnei 76:81 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 0:116 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 1 16 theme rare 205:208 arg1 monosaccharides 210:224 two rare monosaccharides 201:224 two rare monosaccharides 201:224 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 0 17 theme unit 40:43 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 0:116 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 1 18 dep repeat 181:186 arg1 features 144:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 0 19 theme repeating 30:38 arg1 unit 40:43 the zwitterionic repeating unit 13:43 the zwitterionic repeating unit of the O-antigen 13:60 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 3 20 theme oxidation 411:419 arg1 strategy 421:428 a postglycosylation oxidation strategy 391:428 a postglycosylation oxidation strategy in combination with imidate chemistry 391:466 All targets were synthesized using a postglycosylation oxidation strategy in combination with imidate chemistry. 25210812 1 21 theme O-antigen 134:142 arg1 features 144:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 2 22 theme chain 311:315 arg1 elongation 317:326 chain elongation 311:326 chain elongation 311:326 The synthesis of the AB repeat and of trisaccharides ABA' and B'AB, which validates chain elongation at either end, is reported. 25210812 0 23 from elongation 93:102 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 0:116 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 6 24 theme 2/C-mediated 605:616 arg1 deprotection 618:629 A one-step Pd(OH)2/C-mediated deprotection 588:629 A one-step Pd(OH)2/C-mediated deprotection 588:629 A one-step Pd(OH)2/C-mediated deprotection provided the propyl glycoside targets. 25210812 1 25 theme Shigella 118:125 arg1 sonnei 127:132 Shigella sonnei 118:132 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 2 26 theme repeat 251:256 arg1 synthesis 231:239 The synthesis 227:239 The synthesis 227:239 The synthesis of the AB repeat and of trisaccharides ABA' and B'AB, which validates chain elongation at either end, is reported. 25210812 1 27 theme sonnei 127:132 arg1 features 144:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features 118:151 Shigella sonnei O-antigen features a zwitterionic disaccharide repeat encompassing two rare monosaccharides. 25210812 0 28 from ends 112:115 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 0:116 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 0 29 theme O-antigen 52:60 arg1 unit 40:43 the zwitterionic repeating unit 13:43 the zwitterionic repeating unit of the O-antigen 13:60 Synthesis of the zwitterionic repeating unit of the O-antigen from Shigella sonnei and chain elongation at both ends. 25210812 2 30 theme AB 248:249 arg1 repeat 251:256 the AB repeat 244:256 the AB repeat 244:256 The synthesis of the AB repeat and of trisaccharides ABA' and B'AB, which validates chain elongation at either end, is reported. 25210812 2 31 dep trisaccharides 265:278 arg1 AB 291:292 ABA' and B'AB 280:292 trisaccharides ABA' and B'AB 265:292 The synthesis of the AB repeat and of trisaccharides ABA' and B'AB, which validates chain elongation at either end, is reported. 26908049 10 0 theme drug 1698:1701 arg1 development 1703:1713 drug development 1698:1713 drug development of glycoproteins 1698:1730 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 9 1 theme ART 1457:1459 arg1 solubility 1429:1438 solubility 1429:1438 solubility 1429:1438 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 9 1 theme ART 1457:1459 arg1 stability 1444:1452 stability 1444:1452 stability 1444:1452 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 4 2 theme Biological 607:616 arg1 assays 618:623 Biological assays 607:623 Biological assays 607:623 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 5 3 theme glycosylation 798:810 arg1 impact 788:793 the conformational impact 769:793 the conformational impact of glycosylation on ART 769:817 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 9 4 theme colloidal 1350:1358 arg1 stability 1360:1368 thermal and colloidal stability 1338:1368 stability 1360:1368 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 10 5 gly glycoproteins 1718:1730 arg1 glycoproteins 1718:1730 glycoproteins 1718:1730 glycoproteins 1718:1730 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 1 6 theme unexplored 166:175 arg1 scaffold 177:184 a largely unexplored scaffold 156:184 a largely unexplored scaffold for deconstructing and attributing novel functions to proteins 156:247 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 8 7 theme protein 1301:1307 arg1 dynamics 1285:1292 dynamics 1285:1292 dynamics 1285:1292 Surprisingly, the HDX-MS data indicated that the glycan does not greatly influence the conformation and dynamics of the protein. 26908049 8 7 theme protein 1301:1307 arg1 conformation 1268:1279 conformation 1268:1279 conformation 1268:1279 Surprisingly, the HDX-MS data indicated that the glycan does not greatly influence the conformation and dynamics of the protein. 26908049 10 8 theme multiple 1526:1533 arg1 techniques 1544:1553 multiple advanced techniques 1526:1553 multiple advanced techniques 1526:1553 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 7 9 gly nonglycosylated 1160:1174 arg1 ART 1176:1178 glycosylated and nonglycosylated ART 1143:1178 glycosylated and nonglycosylated ART 1143:1178 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 6 10 theme colloidal 942:950 arg1 stability 952:960 The colloidal stability 938:960 The colloidal stability of ART glycovariants 938:981 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 10 11 used used 1562:1565 arg2 analysis 1511:1518 careful analysis 1503:1518 careful analysis using multiple advanced techniques 1503:1553 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 9 12 gly glycosylation 1397:1409 arg1 ART 1457:1459 ART 1457:1459 ART 1457:1459 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 3 13 theme various 590:596 arg1 enzymes 598:604 various enzymes 590:604 various enzymes 590:604 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 10 14 theme careful 1503:1509 arg1 analysis 1511:1518 careful analysis 1503:1518 careful analysis using multiple advanced techniques 1503:1553 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 6 15 theme viscometry 1025:1034 arg1 assays 1052:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 2 16 gly glycoprotein 284:295 arg1 glycoprotein 284:295 the glycoprotein Artemin 280:303 the glycoprotein Artemin (ART) 280:309 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 9 17 from differences 1323:1333 arg1 stability 1360:1368 thermal and colloidal stability 1338:1368 stability 1360:1368 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 2 18 theme N-linked 427:434 arg1 glycosylation 436:448 N-linked glycosylation 427:448 N-linked glycosylation 427:448 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 1 19 theme diverse 79:85 arg1 roles 116:120 diverse and unpredictable functional roles 79:120 diverse and unpredictable functional roles 79:120 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 7 20 gly glycosylated 1143:1154 arg1 ART 1176:1178 glycosylated and nonglycosylated ART 1143:1178 glycosylated and nonglycosylated ART 1143:1178 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 4 21 theme pharmacokinetic 684:698 arg1 function 723:730 a function 721:730 a function of glycosylation 721:747 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 21 theme pharmacokinetic 684:698 arg1 properties 700:709 pharmacokinetic properties 684:709 pharmacokinetic properties of ART 684:716 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 21 theme pharmacokinetic 684:698 arg1 activity 671:678 the relative activity 658:678 the relative activity 658:678 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 5 22 theme hydrogen/deuterium 820:837 arg1 HDX-MS 867:872 HDX-MS 867:872 HDX-MS 867:872 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 5 22 theme hydrogen/deuterium 820:837 arg1 spectrometry 853:864 hydrogen/deuterium exchange mass spectrometry 820:864 hydrogen/deuterium exchange mass spectrometry (HDX-MS) 820:873 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 2 23 theme ART 501:503 arg1 properties 487:496 the structural and functional properties 457:496 the structural and functional properties of ART 457:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 2 24 theme glycoprotein 284:295 arg1 ART 306:308 ART 306:308 ART 306:308 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 2 24 theme glycoprotein 284:295 arg1 Artemin 297:303 the glycoprotein Artemin 280:303 the glycoprotein Artemin (ART) 280:309 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 10 25 theme glycosylation 1635:1647 arg1 functions 1614:1622 the multiple potential functions 1591:1622 the multiple potential functions of protein glycosylation 1591:1647 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 2 26 theme comprehensive 392:404 arg1 understanding 406:418 a comprehensive understanding 390:418 a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART 390:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 3 27 with incubation 574:583 arg1 enzymes 598:604 various enzymes 590:604 various enzymes 590:604 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 9 28 dep solubility 1429:1438 arg1 the 1425:1427 the 1425:1427 the 1425:1427 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 8 29 theme HDX-MS 1199:1204 arg1 data 1206:1209 the HDX-MS data 1195:1209 the HDX-MS data 1195:1209 Surprisingly, the HDX-MS data indicated that the glycan does not greatly influence the conformation and dynamics of the protein. 26908049 1 30 theme unpredictable 91:103 arg1 roles 116:120 diverse and unpredictable functional roles 79:120 diverse and unpredictable functional roles 79:120 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 3 31 theme ART 553:555 arg1 glycan 543:548 the N-linked glycan 530:548 the N-linked glycan of ART 530:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 1 32 theme deconstructing 190:203 arg1 functions 227:235 deconstructing and attributing novel functions 190:235 deconstructing and attributing novel functions 190:235 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 2 33 theme structural 461:470 arg1 properties 487:496 the structural and functional properties 457:496 the structural and functional properties of ART 457:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 5 34 from impact 788:793 arg1 ART 815:817 ART 815:817 ART 815:817 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 1 35 theme functional 105:114 arg1 roles 116:120 diverse and unpredictable functional roles 79:120 diverse and unpredictable functional roles 79:120 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 6 36 theme solubility 1041:1050 arg1 assays 1052:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 5 37 theme differential 903:914 arg1 calorimetry 925:935 differential scanning calorimetry 903:935 differential scanning calorimetry 903:935 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 5 38 theme mass 848:851 arg1 HDX-MS 867:872 HDX-MS 867:872 HDX-MS 867:872 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 5 38 theme mass 848:851 arg1 spectrometry 853:864 hydrogen/deuterium exchange mass spectrometry 820:864 hydrogen/deuterium exchange mass spectrometry (HDX-MS) 820:873 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 4 39 used used 642:645 arg2 systems 629:635 systems 629:635 systems 629:635 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 39 used used 642:645 arg2 assays 618:623 Biological assays 607:623 Biological assays 607:623 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 10 40 theme advanced 1535:1542 arg1 techniques 1544:1553 multiple advanced techniques 1526:1553 multiple advanced techniques 1526:1553 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 1 41 theme attributing 209:219 arg1 functions 227:235 deconstructing and attributing novel functions 190:235 deconstructing and attributing novel functions 190:235 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 6 42 theme light 1007:1011 arg1 scattering 1013:1022 dynamic light scattering 999:1022 dynamic light scattering 999:1022 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 3 43 link N-linked 534:541 arg1 glycan 543:548 the N-linked glycan 530:548 the N-linked glycan of ART 530:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 3 44 theme N-linked 534:541 arg1 glycan 543:548 the N-linked glycan 530:548 the N-linked glycan of ART 530:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 1 45 theme novel 221:225 arg1 functions 227:235 deconstructing and attributing novel functions 190:235 deconstructing and attributing novel functions 190:235 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 0 46 theme Glycosylation 34:46 arg1 Role 18:21 the Role 14:21 the Role of Artemin Glycosylation 14:46 Investigating the Role of Artemin Glycosylation. 26908049 6 47 theme dynamic 999:1005 arg1 scattering 1013:1022 dynamic light scattering 999:1022 dynamic light scattering 999:1022 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 7 48 theme relative 1105:1112 arg1 potency 1114:1120 relative potency 1105:1120 relative potency 1105:1120 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 10 49 dep CONCLUSIONS 1462:1472 arg1 demonstrate 1487:1497 demonstrate 1487:1497 demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins 1487:1730 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 4 50 theme relative 662:669 arg1 properties 700:709 pharmacokinetic properties 684:709 pharmacokinetic properties of ART 684:716 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 50 theme relative 662:669 arg1 activity 671:678 the relative activity 658:678 the relative activity 658:678 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 50 theme relative 662:669 arg1 function 723:730 a function 721:730 a function of glycosylation 721:747 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 0 51 theme Artemin 26:32 arg1 Glycosylation 34:46 Artemin Glycosylation 26:46 Artemin Glycosylation 26:46 Investigating the Role of Artemin Glycosylation. 26908049 8 52 dep conformation 1268:1279 arg1 the 1264:1266 the 1264:1266 the 1264:1266 Surprisingly, the HDX-MS data indicated that the glycan does not greatly influence the conformation and dynamics of the protein. 26908049 9 53 theme glycosylation 1397:1409 arg1 role 1389:1392 a role 1387:1392 a role of glycosylation in increasing the solubility and stability of ART 1387:1459 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 7 54 theme RESULTS 1060:1066 arg1 difference 1071:1080 RESULTS No difference 1060:1080 RESULTS No difference in pharmacokinetics or relative potency 1060:1120 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 3 55 mod Modification 514:525 arg3 METHODS 506:512 METHODS Modification 506:525 METHODS Modification of the N-linked glycan of ART 506:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 3 55 mod Modification 514:525 arg1 glycan 543:548 the N-linked glycan 530:548 the N-linked glycan of ART 530:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 5 56 theme scanning 916:923 arg1 calorimetry 925:935 differential scanning calorimetry 903:935 differential scanning calorimetry 903:935 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 10 57 theme potential 1604:1612 arg1 functions 1614:1622 the multiple potential functions 1591:1622 the multiple potential functions of protein glycosylation 1591:1647 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 6 58 theme scattering 1013:1022 arg1 assays 1052:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 dynamic light scattering, viscometry, and solubility assays 999:1057 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 7 59 theme No 1068:1069 arg1 difference 1071:1080 RESULTS No difference 1060:1080 RESULTS No difference in pharmacokinetics or relative potency 1060:1120 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 4 60 theme glycosylation 735:747 arg1 function 723:730 a function 721:730 a function of glycosylation 721:747 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 60 theme glycosylation 735:747 arg1 properties 700:709 pharmacokinetic properties 684:709 pharmacokinetic properties of ART 684:716 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 60 theme glycosylation 735:747 arg1 activity 671:678 the relative activity 658:678 the relative activity 658:678 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 2 61 theme functional 476:485 arg1 properties 487:496 the structural and functional properties 457:496 the structural and functional properties of ART 457:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 9 62 theme thermal 1338:1344 arg1 stability 1360:1368 thermal and colloidal stability 1338:1368 stability 1360:1368 In contrast, differences in thermal and colloidal stability clearly revealed a role of glycosylation in increasing the solubility and stability of ART. 26908049 7 63 theme nonglycosylated 1160:1174 arg1 ART 1176:1178 glycosylated and nonglycosylated ART 1143:1178 glycosylated and nonglycosylated ART 1143:1178 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 2 64 theme analytical 346:355 arg1 methods 357:363 multiple analytical methods 337:363 multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART 337:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 6 65 theme glycovariants 969:981 arg1 stability 952:960 The colloidal stability 938:960 The colloidal stability of ART glycovariants 938:981 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 2 66 theme multiple 337:344 arg1 methods 357:363 multiple analytical methods 337:363 multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART 337:503 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 7 67 from difference 1071:1080 arg1 pharmacokinetics 1085:1100 pharmacokinetics 1085:1100 pharmacokinetics 1085:1100 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 7 67 from difference 1071:1080 arg1 potency 1114:1120 relative potency 1105:1120 relative potency 1105:1120 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 5 68 theme exchange 839:846 arg1 HDX-MS 867:872 HDX-MS 867:872 HDX-MS 867:872 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 5 68 theme exchange 839:846 arg1 spectrometry 853:864 hydrogen/deuterium exchange mass spectrometry 820:864 hydrogen/deuterium exchange mass spectrometry (HDX-MS) 820:873 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 3 69 theme METHODS 506:512 arg1 Modification 514:525 METHODS Modification 506:525 METHODS Modification of the N-linked glycan of ART 506:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 7 70 theme glycosylated 1143:1154 arg1 ART 1176:1178 glycosylated and nonglycosylated ART 1143:1178 glycosylated and nonglycosylated ART 1143:1178 RESULTS No difference in pharmacokinetics or relative potency was revealed between glycosylated and nonglycosylated ART. 26908049 5 71 theme conformational 773:786 arg1 impact 788:793 the conformational impact 769:793 the conformational impact of glycosylation on ART 769:817 In order to reveal the conformational impact of glycosylation on ART, hydrogen/deuterium exchange mass spectrometry (HDX-MS) was employed in addition to differential scanning calorimetry. 26908049 2 72 link N-linked 427:434 arg1 glycosylation 436:448 N-linked glycosylation 427:448 N-linked glycosylation 427:448 Here, the glycoprotein Artemin (ART) was carefully assessed by multiple analytical methods that allow us to provide a comprehensive understanding of how N-linked glycosylation impact the structural and functional properties of ART. 26908049 3 73 theme glycan 543:548 arg1 Modification 514:525 METHODS Modification 506:525 METHODS Modification of the N-linked glycan of ART 506:555 METHODS Modification of the N-linked glycan of ART was performed by incubation with various enzymes. 26908049 10 74 theme multiple 1595:1602 arg1 functions 1614:1622 the multiple potential functions 1591:1622 the multiple potential functions of protein glycosylation 1591:1647 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 1 75 theme drug 256:259 arg1 development 261:271 drug development 256:271 drug development 256:271 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 10 76 theme protein 1627:1633 arg1 glycosylation 1635:1647 protein glycosylation 1627:1647 protein glycosylation 1627:1647 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 6 77 theme ART 965:967 arg1 glycovariants 969:981 ART glycovariants 965:981 ART glycovariants 965:981 The colloidal stability of ART glycovariants was assessed by dynamic light scattering, viscometry, and solubility assays. 26908049 1 78 theme PURPOSE 49:55 arg1 Oligosaccharides 57:72 PURPOSE Oligosaccharides 49:72 PURPOSE Oligosaccharides 49:72 PURPOSE Oligosaccharides play diverse and unpredictable functional roles when attached to proteins and are a largely unexplored scaffold for deconstructing and attributing novel functions to proteins during drug development. 26908049 4 79 theme ART 714:716 arg1 activity 671:678 the relative activity 658:678 the relative activity 658:678 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 79 theme ART 714:716 arg1 properties 700:709 pharmacokinetic properties 684:709 pharmacokinetic properties of ART 684:716 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 4 79 theme ART 714:716 arg1 function 723:730 a function 721:730 a function of glycosylation 721:747 Biological assays and systems were used to examine the relative activity and pharmacokinetic properties of ART as a function of glycosylation. 26908049 10 80 theme glycoproteins 1718:1730 arg1 glycoproteins 1718:1730 glycoproteins 1718:1730 glycoproteins 1718:1730 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 10 80 theme glycoproteins 1718:1730 arg1 development 1703:1713 drug development 1698:1713 drug development of glycoproteins 1698:1730 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 26908049 10 80 theme glycoproteins 1718:1730 arg1 prerequisite 1660:1671 a prerequisite 1658:1671 a prerequisite for glycoengineering 1658:1692 CONCLUSIONS Our findings demonstrate how careful analysis using multiple advanced techniques can be used to identify and dissect the multiple potential functions of protein glycosylation and form a prerequisite for glycoengineering and drug development of glycoproteins. 28323990 12 0 gly glycosylation 1900:1912 arg1 treatment 1989:1997 successful dietary treatment 1970:1997 successful dietary treatment in TMEM165 deficiency 1970:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 0 gly glycosylation 1900:1912 arg1 lipids 1917:1922 lipids 1917:1922 lipids in the TMEM165 defect 1917:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 11 1 theme transferrin 1695:1705 arg1 isoforms 1707:1714 the negatively charged transferrin isoforms 1672:1714 the negatively charged transferrin isoforms 1672:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 10 2 theme observed 1409:1416 arg1 defect 1434:1439 the observed N-glycosylation defect 1405:1439 the observed N-glycosylation defect 1405:1439 Although these defects were completely corrected by the addition of Mn2+, we demonstrated that the observed N-glycosylation defect could also be overcome by galactose supplementation. 28323990 11 3 theme TMEM165-deficient 1568:1584 arg1 CDG 1586:1588 TMEM165-deficient CDG 1568:1588 TMEM165-deficient CDG 1568:1588 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 6 4 from glycosylation 699:711 arg1 cells 740:744 TMEM165-depleted HEK293 cells 716:744 TMEM165-depleted HEK293 cells 716:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 6 5 from effects 653:659 arg1 patients 763:770 2 patients 761:770 2 patients with TMEM165-CDG 761:787 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 6 5 from effects 653:659 arg1 cells 827:831 their cultured skin fibroblast cells 796:831 their cultured skin fibroblast cells 796:831 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 6 5 from effects 653:659 arg1 glycosylation 699:711 Golgi glycosylation 693:711 Golgi glycosylation in TMEM165-depleted HEK293 cells 693:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 9 6 theme knockout 1213:1220 arg1 cells 1237:1241 knockout TMEM165 HEK293 cells 1213:1241 knockout TMEM165 HEK293 cells 1213:1241 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 2 7 theme growing 296:302 arg1 diseases 323:330 inherited diseases 313:330 inherited diseases in which the glycosylation process is altered 313:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 7 theme growing 296:302 arg1 type 233:236 one type 229:236 one type of congenital disorders of glycosylation (CDG) 229:283 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 7 theme growing 296:302 arg1 group 304:308 a rapidly growing group 286:308 a rapidly growing group of inherited diseases in which the glycosylation process is altered 286:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 1 8 theme skeletal 195:202 arg1 involvement 204:214 metabolic, endocrine, and skeletal involvement 169:214 metabolic, endocrine, and skeletal involvement 169:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 5 9 theme CDG 566:568 arg1 types 570:574 other CDG types 560:574 other CDG types with abnormal galactosylation 560:604 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 10 10 theme N-glycosylation 1418:1432 arg1 defect 1434:1439 the observed N-glycosylation defect 1405:1439 the observed N-glycosylation defect 1405:1439 Although these defects were completely corrected by the addition of Mn2+, we demonstrated that the observed N-glycosylation defect could also be overcome by galactose supplementation. 28323990 9 11 theme HEK293 1230:1235 arg1 cells 1237:1241 knockout TMEM165 HEK293 cells 1213:1241 knockout TMEM165 HEK293 cells 1213:1241 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 5 12 from beneficial 546:555 arg1 types 570:574 other CDG types 560:574 other CDG types with abnormal galactosylation 560:604 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 1 13 theme multisystem 129:139 arg1 disease 141:147 a severe multisystem disease 120:147 a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement 120:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 1 13 theme multisystem 129:139 arg1 deficiency 106:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 12 14 dep CONCLUSION 1829:1838 arg1 this 1858:1861 this 1858:1861 this 1858:1861 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 14 dep CONCLUSION 1829:1838 arg1 description 1876:1886 the first description 1866:1886 the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency 1866:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 11 15 from increase 1660:1667 arg1 structures 1769:1778 hypogalactosylated total N-glycan structures 1735:1778 hypogalactosylated total N-glycan structures 1735:1778 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 15 from increase 1660:1667 arg1 function 1791:1798 endocrine function 1781:1798 endocrine function 1781:1798 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 15 from increase 1660:1667 arg1 isoforms 1707:1714 the negatively charged transferrin isoforms 1672:1714 the negatively charged transferrin isoforms 1672:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 15 from increase 1660:1667 arg1 parameters 1817:1826 coagulation parameters 1805:1826 coagulation parameters 1805:1826 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 6 16 with patients 763:770 arg1 TMEM165-CDG 777:787 TMEM165-CDG 777:787 TMEM165-CDG 777:787 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 9 17 theme severe 1254:1259 arg1 defects 1301:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 5 18 from types 570:574 arg1 beneficial 546:555 beneficial 546:555 beneficial 546:555 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 9 19 theme N-linked 1177:1184 arg1 glycans 1186:1192 N-linked glycans 1177:1192 N-linked glycans 1177:1192 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 11 20 theme galactose 1525:1533 arg1 supplementation 1535:1549 oral galactose supplementation 1520:1549 oral galactose supplementation in patients with TMEM165-deficient CDG 1520:1588 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 5 21 theme OBJECTIVE 505:513 arg1 Galactose 515:523 OBJECTIVE Galactose 505:523 OBJECTIVE Galactose 505:523 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 6 22 theme study 623:627 arg1 aim 611:613 The aim 607:613 The aim of this study 607:627 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 11 23 theme coagulation 1805:1815 arg1 parameters 1817:1826 coagulation parameters 1805:1826 coagulation parameters 1805:1826 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 0 24 theme Glycosylation 67:79 arg1 Defects 81:87 the Glycosylation Defects 63:87 the Glycosylation Defects 63:87 Galactose Supplementation in Patients With TMEM165-CDG Rescues the Glycosylation Defects. 28323990 9 25 theme transfer 1292:1299 arg1 defects 1301:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 2 26 theme congenital 241:250 arg1 disorders 252:260 congenital disorders 241:260 congenital disorders of glycosylation (CDG) 241:283 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 8 27 with INTERVENTIONS 974:986 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 27 with INTERVENTIONS 974:986 arg1 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 27 with INTERVENTIONS 974:986 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 10 28 theme galactose 1467:1475 arg1 supplementation 1477:1491 galactose supplementation 1467:1491 galactose supplementation 1467:1491 Although these defects were completely corrected by the addition of Mn2+, we demonstrated that the observed N-glycosylation defect could also be overcome by galactose supplementation. 28323990 12 29 theme dietary 1981:1987 arg1 treatment 1989:1997 successful dietary treatment 1970:1997 successful dietary treatment in TMEM165 deficiency 1970:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 13 30 from use 2039:2041 arg1 TMEM165-CDG 2074:2084 TMEM165-CDG 2074:2084 TMEM165-CDG 2074:2084 We recommend the use of oral d-galactose therapy in TMEM165-CDG. 28323990 8 31 theme deep 1043:1046 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 31 theme deep 1043:1046 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 7 32 theme transferrin 929:939 arg1 isoelectrofocusing 941:958 transferrin isoelectrofocusing 929:958 transferrin isoelectrofocusing 929:958 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 6 33 theme Golgi 693:697 arg1 glycosylation 699:711 Golgi glycosylation 693:711 Golgi glycosylation in TMEM165-depleted HEK293 cells 693:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 8 34 theme homozygous 1057:1066 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 34 theme homozygous 1057:1066 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 1 35 theme CONTEXT 90:96 arg1 disease 141:147 a severe multisystem disease 120:147 a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement 120:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 1 35 theme CONTEXT 90:96 arg1 deficiency 106:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 4 36 theme only 470:473 arg1 types 484:488 only specific types 470:488 only specific types 470:488 There are >100 CDGs, but only specific types are treatable. 28323990 6 37 theme supplementation 674:688 arg1 effects 653:659 the effects 649:659 the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells 649:831 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 3 38 theme serum 422:426 arg1 analysis 435:442 serum glycan analysis 422:442 serum glycan analysis 422:442 Patients have decreased galactosylation by serum glycan analysis. 28323990 0 39 theme Galactose 0:8 arg1 Supplementation 10:24 Galactose Supplementation 0:24 Galactose Supplementation in Patients With TMEM165-CDG 0:53 Galactose Supplementation in Patients With TMEM165-CDG Rescues the Glycosylation Defects. 28323990 7 40 theme blot 910:913 arg1 analysis 915:922 western blot analysis 902:922 western blot analysis 902:922 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 9 41 dep RESULTS 1157:1163 arg1 analyzed 1168:1175 analyzed 1168:1175 analyzed 1168:1175 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 12 42 theme TMEM165 1931:1937 arg1 defect 1939:1944 the TMEM165 defect 1927:1944 the TMEM165 defect 1927:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 8 43 with PATIENTS 961:968 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 43 with PATIENTS 961:968 arg1 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 43 with PATIENTS 961:968 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 11 44 theme N-glycan 1760:1767 arg1 structures 1769:1778 hypogalactosylated total N-glycan structures 1735:1778 hypogalactosylated total N-glycan structures 1735:1778 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 12 45 theme glycosylation 1900:1912 arg1 this 1858:1861 this 1858:1861 this 1858:1861 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 45 theme glycosylation 1900:1912 arg1 description 1876:1886 the first description 1866:1886 the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency 1866:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 2 46 theme inherited 313:321 arg1 diseases 323:330 inherited diseases 313:330 inherited diseases in which the glycosylation process is altered 313:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 7 47 theme western 902:908 arg1 analysis 915:922 western blot analysis 902:922 western blot analysis 902:922 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 6 48 theme skin 811:814 arg1 cells 827:831 their cultured skin fibroblast cells 796:831 their cultured skin fibroblast cells 796:831 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 0 49 with Patients 29:36 arg1 TMEM165-CDG 43:53 TMEM165-CDG 43:53 TMEM165-CDG 43:53 Galactose Supplementation in Patients With TMEM165-CDG Rescues the Glycosylation Defects. 28323990 11 50 theme charged 1687:1693 arg1 isoforms 1707:1714 the negatively charged transferrin isoforms 1672:1714 the negatively charged transferrin isoforms 1672:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 8 51 theme daily 1135:1139 arg1 dose 1141:1144 a daily dose 1133:1144 a daily dose of 1 g/kg 1133:1154 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 12 52 theme first 1870:1874 arg1 this 1858:1861 this 1858:1861 this 1858:1861 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 52 theme first 1870:1874 arg1 description 1876:1886 the first description 1866:1886 the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency 1866:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 53 theme treatment 1989:1997 arg1 report 1960:1965 the first report 1950:1965 the first report of successful dietary treatment in TMEM165 deficiency 1950:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 53 theme treatment 1989:1997 arg1 glycosylation 1900:1912 abnormal glycosylation 1891:1912 abnormal glycosylation of lipids in the TMEM165 defect 1891:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 11 54 theme biochemical 1599:1609 arg1 decrease 1723:1730 a decrease 1721:1730 a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters 1721:1826 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 54 theme biochemical 1599:1609 arg1 increase 1660:1667 a substantial increase 1646:1667 a substantial increase in the negatively charged transferrin isoforms 1646:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 54 theme biochemical 1599:1609 arg1 parameters 1624:1633 biochemical and clinical parameters 1599:1633 biochemical and clinical parameters 1599:1633 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 8 55 theme g/kg 1151:1154 arg1 dose 1141:1144 a daily dose 1133:1144 a daily dose of 1 g/kg 1133:1154 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 56 theme unrelated 993:1001 arg1 patients 1003:1010 unrelated patients 993:1010 unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) 993:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 9 57 link N-linked 1177:1184 arg1 glycans 1186:1192 N-linked glycans 1177:1192 N-linked glycans 1177:1192 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 11 58 theme hypogalactosylated 1735:1752 arg1 structures 1769:1778 hypogalactosylated total N-glycan structures 1735:1778 hypogalactosylated total N-glycan structures 1735:1778 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 13 59 theme oral 2046:2049 arg1 therapy 2063:2069 oral d-galactose therapy 2046:2069 oral d-galactose therapy 2046:2069 We recommend the use of oral d-galactose therapy in TMEM165-CDG. 28323990 7 60 theme SETTING 845:851 arg1 Glycosylation 853:865 DESIGN AND SETTING Glycosylation 834:865 DESIGN AND SETTING Glycosylation 834:865 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 1 61 theme endocrine 180:188 arg1 involvement 204:214 metabolic, endocrine, and skeletal involvement 169:214 metabolic, endocrine, and skeletal involvement 169:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 7 62 theme DESIGN 834:839 arg1 Glycosylation 853:865 DESIGN AND SETTING Glycosylation 834:865 DESIGN AND SETTING Glycosylation 834:865 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 11 63 with patients 1554:1561 arg1 CDG 1586:1588 TMEM165-deficient CDG 1568:1588 TMEM165-deficient CDG 1568:1588 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 64 theme clinical 1615:1622 arg1 decrease 1723:1730 a decrease 1721:1730 a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters 1721:1826 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 64 theme clinical 1615:1622 arg1 increase 1660:1667 a substantial increase 1646:1667 a substantial increase in the negatively charged transferrin isoforms 1646:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 64 theme clinical 1615:1622 arg1 parameters 1624:1633 biochemical and clinical parameters 1599:1633 biochemical and clinical parameters 1599:1633 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 12 65 from defect 1939:1944 arg1 report 1960:1965 the first report 1950:1965 the first report of successful dietary treatment in TMEM165 deficiency 1950:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 65 from defect 1939:1944 arg1 glycosylation 1900:1912 abnormal glycosylation 1891:1912 abnormal glycosylation of lipids in the TMEM165 defect 1891:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 13 66 theme therapy 2063:2069 arg1 use 2039:2041 the use 2035:2041 the use of oral d-galactose therapy in TMEM165-CDG 2035:2084 We recommend the use of oral d-galactose therapy in TMEM165-CDG. 28323990 5 67 with types 570:574 arg1 galactosylation 590:604 abnormal galactosylation 581:604 abnormal galactosylation 581:604 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 1 68 theme severe 122:127 arg1 disease 141:147 a severe multisystem disease 120:147 a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement 120:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 1 68 theme severe 122:127 arg1 deficiency 106:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 12 69 from lipids 1917:1922 arg1 defect 1939:1944 the TMEM165 defect 1927:1944 the TMEM165 defect 1927:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 6 70 gly glycosylation 699:711 arg1 cells 740:744 TMEM165-depleted HEK293 cells 716:744 TMEM165-depleted HEK293 cells 716:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 12 71 theme TMEM165 2002:2008 arg1 deficiency 2010:2019 TMEM165 deficiency 2002:2019 TMEM165 deficiency 2002:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 11 72 theme oral 1520:1523 arg1 supplementation 1535:1549 oral galactose supplementation 1520:1549 oral galactose supplementation in patients with TMEM165-deficient CDG 1520:1588 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 9 73 theme TMEM165 1222:1228 arg1 cells 1237:1241 knockout TMEM165 HEK293 cells 1213:1241 knockout TMEM165 HEK293 cells 1213:1241 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 9 74 theme hypogalactosylation 1261:1279 arg1 defects 1301:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 2 75 theme glycosylation 265:277 arg1 disorders 252:260 congenital disorders 241:260 congenital disorders of glycosylation (CDG) 241:283 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 9 76 theme GalNAc 1285:1290 arg1 defects 1301:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 severe hypogalactosylation and GalNAc transfer defects 1254:1307 RESULTS We analyzed N-linked glycans and glycolipids in knockout TMEM165 HEK293 cells, revealing severe hypogalactosylation and GalNAc transfer defects. 28323990 2 77 theme disorders 252:260 arg1 type 233:236 one type 229:236 one type of congenital disorders of glycosylation (CDG) 229:283 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 77 theme disorders 252:260 arg1 diseases 323:330 inherited diseases 313:330 inherited diseases in which the glycosylation process is altered 313:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 77 theme disorders 252:260 arg1 group 304:308 a rapidly growing group 286:308 a rapidly growing group of inherited diseases in which the glycosylation process is altered 286:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 12 78 from treatment 1989:1997 arg1 deficiency 2010:2019 TMEM165 deficiency 2002:2019 TMEM165 deficiency 2002:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 8 79 theme same 1038:1041 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 79 theme same 1038:1041 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 1 80 theme metabolic 169:177 arg1 involvement 204:214 metabolic, endocrine, and skeletal involvement 169:214 metabolic, endocrine, and skeletal involvement 169:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 0 81 from Supplementation 10:24 arg1 Patients 29:36 Patients 29:36 Patients With TMEM165-CDG 29:53 Galactose Supplementation in Patients With TMEM165-CDG Rescues the Glycosylation Defects. 28323990 8 82 theme intronic 1048:1055 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 82 theme intronic 1048:1055 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 5 83 theme abnormal 581:588 arg1 galactosylation 590:604 abnormal galactosylation 581:604 abnormal galactosylation 581:604 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 11 84 from supplementation 1535:1549 arg1 patients 1554:1561 patients 1554:1561 patients with TMEM165-deficient CDG 1554:1588 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 12 85 from report 1960:1965 arg1 defect 1939:1944 the TMEM165 defect 1927:1944 the TMEM165 defect 1927:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 85 from report 1960:1965 arg1 deficiency 2010:2019 TMEM165 deficiency 2002:2019 TMEM165 deficiency 2002:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 86 theme report 1960:1965 arg1 this 1858:1861 this 1858:1861 this 1858:1861 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 86 theme report 1960:1965 arg1 description 1876:1886 the first description 1866:1886 the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency 1866:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 8 87 with patients 1003:1010 arg1 mutation 1068:1075 the same deep intronic homozygous mutation 1034:1075 the same deep intronic homozygous mutation (c.792+182G>A) 1034:1090 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 87 with patients 1003:1010 arg1 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 TMEM165-CDG 1017:1027 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 8 87 with patients 1003:1010 arg1 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 c.792+182G>A 1078:1089 PATIENTS AND INTERVENTIONS Both unrelated patients with TMEM165-CDG with the same deep intronic homozygous mutation (c.792+182G>A) were allocated to receive d-galactose in a daily dose of 1 g/kg. 28323990 4 88 theme specific 475:482 arg1 types 484:488 only specific types 470:488 only specific types 470:488 There are >100 CDGs, but only specific types are treatable. 28323990 12 89 theme successful 1970:1979 arg1 treatment 1989:1997 successful dietary treatment 1970:1997 successful dietary treatment in TMEM165 deficiency 1970:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 5 90 theme other 560:564 arg1 types 570:574 other CDG types 560:574 other CDG types with abnormal galactosylation 560:604 OBJECTIVE Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation. 28323990 1 91 theme TMEM165 98:104 arg1 disease 141:147 a severe multisystem disease 120:147 a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement 120:214 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 1 91 theme TMEM165 98:104 arg1 deficiency 106:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency 90:115 CONTEXT TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine, and skeletal involvement. 28323990 11 92 from decrease 1723:1730 arg1 structures 1769:1778 hypogalactosylated total N-glycan structures 1735:1778 hypogalactosylated total N-glycan structures 1735:1778 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 92 from decrease 1723:1730 arg1 function 1791:1798 endocrine function 1781:1798 endocrine function 1781:1798 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 92 from decrease 1723:1730 arg1 parameters 1817:1826 coagulation parameters 1805:1826 coagulation parameters 1805:1826 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 92 from decrease 1723:1730 arg1 isoforms 1707:1714 the negatively charged transferrin isoforms 1672:1714 the negatively charged transferrin isoforms 1672:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 12 93 from deficiency 2010:2019 arg1 report 1960:1965 the first report 1950:1965 the first report of successful dietary treatment in TMEM165 deficiency 1950:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 93 from deficiency 2010:2019 arg1 glycosylation 1900:1912 abnormal glycosylation 1891:1912 abnormal glycosylation of lipids in the TMEM165 defect 1891:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 6 94 theme galactose 664:672 arg1 supplementation 674:688 galactose supplementation 664:688 galactose supplementation 664:688 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 3 95 theme glycan 428:433 arg1 analysis 435:442 serum glycan analysis 422:442 serum glycan analysis 422:442 Patients have decreased galactosylation by serum glycan analysis. 28323990 6 96 theme HEK293 733:738 arg1 cells 740:744 TMEM165-depleted HEK293 cells 716:744 TMEM165-depleted HEK293 cells 716:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 11 97 theme total 1754:1758 arg1 structures 1769:1778 hypogalactosylated total N-glycan structures 1735:1778 hypogalactosylated total N-glycan structures 1735:1778 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 11 98 theme substantial 1648:1658 arg1 increase 1660:1667 a substantial increase 1646:1667 a substantial increase in the negatively charged transferrin isoforms 1646:1714 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 12 99 theme lipids 1917:1922 arg1 report 1960:1965 the first report 1950:1965 the first report of successful dietary treatment in TMEM165 deficiency 1950:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 99 theme lipids 1917:1922 arg1 glycosylation 1900:1912 abnormal glycosylation 1891:1912 abnormal glycosylation of lipids in the TMEM165 defect 1891:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 100 from glycosylation 1900:1912 arg1 defect 1939:1944 the TMEM165 defect 1927:1944 the TMEM165 defect 1927:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 100 from glycosylation 1900:1912 arg1 deficiency 2010:2019 TMEM165 deficiency 2002:2019 TMEM165 deficiency 2002:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 12 101 theme first 1954:1958 arg1 report 1960:1965 the first report 1950:1965 the first report of successful dietary treatment in TMEM165 deficiency 1950:2019 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 2 102 theme diseases 323:330 arg1 diseases 323:330 inherited diseases 313:330 inherited diseases in which the glycosylation process is altered 313:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 102 theme diseases 323:330 arg1 type 233:236 one type 229:236 one type of congenital disorders of glycosylation (CDG) 229:283 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 2 102 theme diseases 323:330 arg1 group 304:308 a rapidly growing group 286:308 a rapidly growing group of inherited diseases in which the glycosylation process is altered 286:376 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 28323990 6 103 theme fibroblast 816:825 arg1 cells 827:831 their cultured skin fibroblast cells 796:831 their cultured skin fibroblast cells 796:831 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 12 104 theme abnormal 1891:1898 arg1 glycosylation 1900:1912 abnormal glycosylation 1891:1912 abnormal glycosylation of lipids in the TMEM165 defect 1891:1944 CONCLUSION To our knowledge, this is the first description of abnormal glycosylation of lipids in the TMEM165 defect and the first report of successful dietary treatment in TMEM165 deficiency. 28323990 6 105 theme TMEM165-depleted 716:731 arg1 cells 740:744 TMEM165-depleted HEK293 cells 716:744 TMEM165-depleted HEK293 cells 716:744 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 11 106 theme endocrine 1781:1789 arg1 function 1791:1798 endocrine function 1781:1798 endocrine function 1781:1798 We then demonstrated that oral galactose supplementation in patients with TMEM165-deficient CDG improved biochemical and clinical parameters, including a substantial increase in the negatively charged transferrin isoforms, and a decrease in hypogalactosylated total N-glycan structures, endocrine function, and coagulation parameters. 28323990 13 107 theme d-galactose 2051:2061 arg1 therapy 2063:2069 oral d-galactose therapy 2046:2069 oral d-galactose therapy 2046:2069 We recommend the use of oral d-galactose therapy in TMEM165-CDG. 28323990 7 108 theme mass 883:886 arg1 spectrometry 888:899 mass spectrometry 883:899 mass spectrometry 883:899 DESIGN AND SETTING Glycosylation was assessed by mass spectrometry, western blot analysis, and transferrin isoelectrofocusing. 28323990 6 109 theme cultured 802:809 arg1 cells 827:831 their cultured skin fibroblast cells 796:831 their cultured skin fibroblast cells 796:831 The aim of this study was to characterize the effects of galactose supplementation on Golgi glycosylation in TMEM165-depleted HEK293 cells, as well as in 2 patients with TMEM165-CDG and in their cultured skin fibroblast cells. 28323990 10 110 theme Mn2+ 1378:1381 arg1 addition 1366:1373 the addition 1362:1373 the addition of Mn2+ 1362:1381 Although these defects were completely corrected by the addition of Mn2+, we demonstrated that the observed N-glycosylation defect could also be overcome by galactose supplementation. 28323990 2 111 theme glycosylation 345:357 arg1 process 359:365 the glycosylation process 341:365 the glycosylation process 341:365 It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases in which the glycosylation process is altered. 24798333 0 0 theme variants 90:97 arg1 control 44:50 N-Glycan-dependent and -independent quality control 0:50 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants 0:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 7 1 theme precursors 1289:1298 arg1 interaction 1261:1271 the co-translational interaction 1240:1271 the co-translational interaction of hδOR-Cys(27) precursors with CNX 1240:1307 This was further supported by the fact that inhibiting the co-translational interaction of hδOR-Cys(27) precursors with CNX led to their ERAD. 24798333 3 2 theme wild-type 471:479 arg1 variants 509:516 wild-type and non-N-glycosylated F27C variants 471:516 wild-type and non-N-glycosylated F27C variants 471:516 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 6 3 theme receptors 1000:1008 arg1 turnover 988:995 The turnover 984:995 The turnover of receptors associating with the molecular chaperone calnexin (CNX) 984:1064 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 6 3 theme receptors 1000:1008 arg1 slower 1084:1089 slower 1084:1089 slower 1084:1089 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 0 4 theme N-terminal 79:88 arg1 variants 90:97 human δ opioid receptor N-terminal variants 55:97 human δ opioid receptor N-terminal variants 55:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 1 5 theme ER-associated 234:246 arg1 ERAD 261:264 ERAD 261:264 ERAD 261:264 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 5 theme ER-associated 234:246 arg1 degradation 248:258 ER-associated degradation 234:258 ER-associated degradation (ERAD) 234:265 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 9 6 theme N-glycan-independent 1721:1740 arg1 mechanism 1742:1750 the N-glycan-independent mechanism 1717:1750 the N-glycan-independent mechanism 1717:1750 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 8 7 from retrotranslocation 1503:1520 arg1 variant 1467:1473 the Cys(27) variant 1455:1473 the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol 1455:1535 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 8 8 theme CNX-bound 1384:1392 arg1 receptors 1394:1402 the CNX-bound receptors 1380:1402 the CNX-bound receptors 1380:1402 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 3 9 theme metabolic 521:529 arg1 assays 543:548 metabolic pulse-chase assays 521:548 metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation 521:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 8 10 theme receptor 1335:1342 arg1 chaperones 1360:1369 Opioid receptor pharmacological chaperones 1328:1369 Opioid receptor pharmacological chaperones 1328:1369 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 6 11 theme important 1128:1136 arg1 role 1138:1141 an important role 1125:1141 an important role of CNX in the hδOR N-glycan-dependent QC 1125:1182 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 3 12 theme pulse-chase 531:541 arg1 assays 543:548 metabolic pulse-chase assays 521:548 metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation 521:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 3 13 gly non-N-glycosylated 485:502 arg1 variants 509:516 wild-type and non-N-glycosylated F27C variants 471:516 wild-type and non-N-glycosylated F27C variants 471:516 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 3 14 theme flow 563:566 arg1 cytometry 568:576 flow cytometry 563:576 flow cytometry 563:576 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 7 15 theme co-translational 1244:1259 arg1 interaction 1261:1271 the co-translational interaction 1240:1271 the co-translational interaction of hδOR-Cys(27) precursors with CNX 1240:1307 This was further supported by the fact that inhibiting the co-translational interaction of hδOR-Cys(27) precursors with CNX led to their ERAD. 24798333 9 16 dep alternative 1767:1777 arg1 although 1780:1787 although 1780:1787 although 1780:1787 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 9 16 dep alternative 1767:1777 arg1 accurate 1794:1801 accurate 1794:1801 accurate 1794:1801 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 5 17 theme N-glycan-dependent 820:837 arg1 pathway 839:845 the N-glycan-dependent pathway 816:845 the N-glycan-dependent pathway 816:845 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 3 18 theme variants 509:516 arg1 trafficking 456:466 trafficking 456:466 trafficking 456:466 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 3 18 theme variants 509:516 arg1 biosynthesis 439:450 biosynthesis 439:450 biosynthesis 439:450 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 9 19 theme altered 1886:1892 arg1 conditions 1903:1912 altered cellular conditions 1886:1912 altered cellular conditions 1886:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 4 20 theme acid 664:667 arg1 difference 669:678 the minute one-amino acid difference 643:678 the minute one-amino acid difference between the variants 643:699 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 3 21 from biosynthesis 439:450 arg1 assays 543:548 metabolic pulse-chase assays 521:548 metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation 521:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 5 22 with incompatible 950:961 arg1 binding 975:981 ligand binding 968:981 ligand binding 968:981 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 9 23 theme cellular 1894:1901 arg1 conditions 1903:1912 altered cellular conditions 1886:1912 altered cellular conditions 1886:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 4 24 theme one-amino 654:662 arg1 difference 669:678 the minute one-amino acid difference 643:678 the minute one-amino acid difference between the variants 643:699 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 9 25 theme alternative 1767:1777 arg1 system 1804:1809 an alternative, although less accurate, system 1764:1809 an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions 1764:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 9 26 theme CNX-mediated 1630:1641 arg1 QC 1662:1663 the CNX-mediated N-glycan-dependent QC 1626:1663 the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions 1626:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 8 27 theme Opioid 1328:1333 arg1 receptor 1335:1342 Opioid receptor 1328:1342 Opioid receptor pharmacological chaperones 1328:1369 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 0 28 theme N-Glycan-dependent 0:17 arg1 control 44:50 N-Glycan-dependent and -independent quality control 0:50 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants 0:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 7 29 with interaction 1261:1271 arg1 CNX 1305:1307 CNX 1305:1307 CNX 1305:1307 This was further supported by the fact that inhibiting the co-translational interaction of hδOR-Cys(27) precursors with CNX led to their ERAD. 24798333 3 30 from trafficking 456:466 arg1 assays 543:548 metabolic pulse-chase assays 521:548 metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation 521:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 1 31 theme Quality 100:106 arg1 QC 117:118 QC 117:118 QC 117:118 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 31 theme Quality 100:106 arg1 control 108:114 Quality control 100:114 Quality control (QC) in the endoplasmic reticulum (ER) 100:153 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 32 theme synthesized 173:183 arg1 proteins 185:192 newly synthesized proteins 167:192 newly synthesized proteins 167:192 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 0 33 theme quality 36:42 arg1 control 44:50 N-Glycan-dependent and -independent quality control 0:50 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants 0:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 4 34 theme hδOR-Cys 732:739 arg1 fraction 720:727 a large fraction 712:727 a large fraction of hδOR-Cys(27) 712:743 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 8 35 dep released 1371:1378 arg1 able 1440:1443 able 1440:1443 able 1440:1443 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 5 36 theme N-glycan-independent 767:786 arg1 unable 795:800 unable 795:800 unable 795:800 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 5 36 theme N-glycan-independent 767:786 arg1 QC 788:789 the N-glycan-independent QC 763:789 the N-glycan-independent QC 763:789 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 2 37 theme -independent 381:392 arg1 manners 394:400 both N-glycan-dependent and -independent manners 353:400 both N-glycan-dependent and -independent manners 353:400 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 0 38 theme -independent 23:34 arg1 control 44:50 N-Glycan-dependent and -independent quality control 0:50 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants 0:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 6 39 theme CNX 1146:1148 arg1 role 1138:1141 an important role 1125:1141 an important role of CNX in the hδOR N-glycan-dependent QC 1125:1182 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 5 40 theme incompatible 950:961 arg1 conformation 937:948 conformation 937:948 conformation incompatible with ligand binding 937:981 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 6 41 from role 1138:1141 arg1 QC 1181:1182 the hδOR N-glycan-dependent QC 1153:1182 the hδOR N-glycan-dependent QC 1153:1182 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 3 42 theme non-N-glycosylated 485:502 arg1 variants 509:516 wild-type and non-N-glycosylated F27C variants 471:516 wild-type and non-N-glycosylated F27C variants 471:516 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 8 43 from polyubiquitination 1480:1497 arg1 variant 1467:1473 the Cys(27) variant 1455:1473 the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol 1455:1535 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 9 44 contain has 1670:1672 arg1 QC 1662:1663 the CNX-mediated N-glycan-dependent QC 1626:1663 the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions 1626:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 9 44 contain has 1670:1672 arg2 capacity 1678:1685 the capacity to assist in folding 1674:1706 the capacity to assist in folding 1674:1706 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 9 45 theme N-glycan-dependent 1643:1660 arg1 QC 1662:1663 the CNX-mediated N-glycan-dependent QC 1626:1663 the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions 1626:1912 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 4 46 theme minute 647:652 arg1 difference 669:678 the minute one-amino acid difference 643:678 the minute one-amino acid difference between the variants 643:699 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 3 47 theme cell 582:585 arg1 biotinylation 595:607 cell surface biotinylation 582:607 cell surface biotinylation 582:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 5 48 theme folded 870:875 arg1 27 905:906 27 905:906 27 905:906 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 5 48 theme folded 870:875 arg1 hδOR-Cys 896:903 some incompletely folded non-N-glycosylated hδOR-Cys 852:903 some incompletely folded non-N-glycosylated hδOR-Cys(27) 852:907 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 2 49 theme N-glycan-dependent 358:375 arg1 manners 394:400 both N-glycan-dependent and -independent manners 353:400 both N-glycan-dependent and -independent manners 353:400 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 3 50 theme surface 587:593 arg1 biotinylation 595:607 cell surface biotinylation 582:607 cell surface biotinylation 582:607 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 6 51 theme chaperone 1041:1049 arg1 CNX 1061:1063 CNX 1061:1063 CNX 1061:1063 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 6 51 theme chaperone 1041:1049 arg1 calnexin 1051:1058 the molecular chaperone calnexin 1027:1058 the molecular chaperone calnexin (CNX) 1027:1064 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 0 52 theme δ 61:61 arg1 variants 90:97 human δ opioid receptor N-terminal variants 55:97 human δ opioid receptor N-terminal variants 55:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 5 53 theme non-N-glycosylated 877:894 arg1 27 905:906 27 905:906 27 905:906 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 5 53 theme non-N-glycosylated 877:894 arg1 hδOR-Cys 896:903 some incompletely folded non-N-glycosylated hδOR-Cys 852:903 some incompletely folded non-N-glycosylated hδOR-Cys(27) 852:907 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 5 54 theme ligand 968:973 arg1 binding 975:981 ligand binding 968:981 ligand binding 968:981 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 4 55 theme large 714:718 arg1 fraction 720:727 a large fraction 712:727 a large fraction of hδOR-Cys(27) 712:743 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 0 56 theme human 55:59 arg1 variants 90:97 human δ opioid receptor N-terminal variants 55:97 human δ opioid receptor N-terminal variants 55:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 5 57 gly non-N-glycosylated 877:894 arg1 27 905:906 27 905:906 27 905:906 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 5 57 gly non-N-glycosylated 877:894 arg1 hδOR-Cys 896:903 some incompletely folded non-N-glycosylated hδOR-Cys 852:903 some incompletely folded non-N-glycosylated hδOR-Cys(27) 852:907 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 1 58 dep scrutinizes 155:165 arg1 either 211:216 either 211:216 either 211:216 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 2 59 theme δ-opioid 304:311 arg1 hδOR 323:326 hδOR 323:326 hδOR 323:326 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 2 59 theme δ-opioid 304:311 arg1 receptor 313:320 the human δ-opioid receptor 294:320 the human δ-opioid receptor (hδOR) 294:327 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 1 60 from control 108:114 arg1 reticulum 140:148 the endoplasmic reticulum 124:148 the endoplasmic reticulum (ER) 124:153 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 60 from control 108:114 arg1 ER 151:152 ER 151:152 ER 151:152 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 8 61 theme ER 1407:1408 arg1 export 1410:1415 ER export 1407:1415 ER export 1407:1415 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 2 62 theme human 298:302 arg1 hδOR 323:326 hδOR 323:326 hδOR 323:326 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 2 62 theme human 298:302 arg1 receptor 313:320 the human δ-opioid receptor 294:320 the human δ-opioid receptor (hδOR) 294:327 Here, we demonstrate that the human δ-opioid receptor (hδOR) is subjected to ERQC in both N-glycan-dependent and -independent manners. 24798333 8 63 theme pharmacological 1344:1358 arg1 chaperones 1360:1369 Opioid receptor pharmacological chaperones 1328:1369 Opioid receptor pharmacological chaperones 1328:1369 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 6 64 theme molecular 1031:1039 arg1 CNX 1061:1063 CNX 1061:1063 CNX 1061:1063 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 6 64 theme molecular 1031:1039 arg1 calnexin 1051:1058 the molecular chaperone calnexin 1027:1058 the molecular chaperone calnexin (CNX) 1027:1064 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 3 65 dep biosynthesis 439:450 arg1 the 435:437 the 435:437 the 435:437 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24798333 5 66 theme cell 921:924 arg1 surface 926:932 the cell surface 917:932 the cell surface 917:932 However, the N-glycan-independent QC was unable to compensate the N-glycan-dependent pathway, and some incompletely folded non-N-glycosylated hδOR-Cys(27) reached the cell surface in conformation incompatible with ligand binding. 24798333 9 67 theme folded 1848:1853 arg1 receptors 1855:1863 misfolded/incompletely folded receptors 1825:1863 misfolded/incompletely folded receptors 1825:1863 Taken together, the hδOR appears to rely primarily on the CNX-mediated N-glycan-dependent QC that has the capacity to assist in folding, whereas the N-glycan-independent mechanism constitutes an alternative, although less accurate, system for directing misfolded/incompletely folded receptors to ERAD, possibly in altered cellular conditions. 24798333 0 68 theme receptor 70:77 arg1 variants 90:97 human δ opioid receptor N-terminal variants 55:97 human δ opioid receptor N-terminal variants 55:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 7 69 theme hδOR-Cys 1276:1283 arg1 precursors 1289:1298 hδOR-Cys(27) precursors 1276:1298 hδOR-Cys(27) precursors 1276:1298 This was further supported by the fact that inhibiting the co-translational interaction of hδOR-Cys(27) precursors with CNX led to their ERAD. 24798333 6 70 theme N-glycan-dependent 1162:1179 arg1 QC 1181:1182 the hδOR N-glycan-dependent QC 1153:1182 the hδOR N-glycan-dependent QC 1153:1182 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 4 71 theme QC 615:616 arg1 mechanisms 618:627 Both QC mechanisms 610:627 Both QC mechanisms 610:627 Both QC mechanisms distinguished the minute one-amino acid difference between the variants, targeting a large fraction of hδOR-Cys(27) to ERAD. 24798333 1 72 theme endoplasmic 128:138 arg1 reticulum 140:148 the endoplasmic reticulum 124:148 the endoplasmic reticulum (ER) 124:153 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 72 theme endoplasmic 128:138 arg1 ER 151:152 ER 151:152 ER 151:152 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 1 73 theme ER 221:222 arg1 export 224:229 ER export 221:229 ER export 221:229 Quality control (QC) in the endoplasmic reticulum (ER) scrutinizes newly synthesized proteins and directs them either to ER export or ER-associated degradation (ERAD). 24798333 0 74 theme opioid 63:68 arg1 variants 90:97 human δ opioid receptor N-terminal variants 55:97 human δ opioid receptor N-terminal variants 55:97 N-Glycan-dependent and -independent quality control of human δ opioid receptor N-terminal variants. 24798333 8 75 theme Cys 1459:1461 arg1 variant 1467:1473 the Cys(27) variant 1455:1473 the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol 1455:1535 Opioid receptor pharmacological chaperones released the CNX-bound receptors to ER export and, furthermore, were able to rescue the Cys(27) variant from polyubiquitination and retrotranslocation to the cytosol whether carrying N-glycans or not. 24798333 6 76 theme hδOR 1157:1160 arg1 QC 1181:1182 the hδOR N-glycan-dependent QC 1153:1182 the hδOR N-glycan-dependent QC 1153:1182 The turnover of receptors associating with the molecular chaperone calnexin (CNX) was significantly slower for the hδOR-Cys(27), pointing to an important role of CNX in the hδOR N-glycan-dependent QC. 24798333 3 77 theme F27C 504:507 arg1 variants 509:516 wild-type and non-N-glycosylated F27C variants 471:516 wild-type and non-N-glycosylated F27C variants 471:516 This was shown by investigating the biosynthesis and trafficking of wild-type and non-N-glycosylated F27C variants in metabolic pulse-chase assays coupled with flow cytometry and cell surface biotinylation. 24028868 8 0 theme Significant 1112:1122 arg1 p 1137:1137 p ≤ 0.05 1137:1144 p ≤ 0.05 1137:1144 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 8 0 theme Significant 1112:1122 arg1 differences 1124:1134 Significant differences 1112:1134 Significant differences (p ≤ 0.05) 1112:1145 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 7 1 theme inflammatory 1079:1090 arg1 proteins 1102:1109 inflammatory mediating proteins 1079:1109 inflammatory mediating proteins 1079:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 10 2 theme = 1482:1482 arg1 0.62 1484:1487 R(2)X(cum) = 0.62 1471:1487 Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62 1418:1487 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 11 3 theme prior 1767:1771 arg1 genders 1759:1765 genders 1759:1765 genders prior to the onset of AD 1759:1790 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 4 4 theme cognitive 574:582 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 7 5 theme those 1070:1074 arg1 groups 1011:1016 the patients groups 998:1016 the patients groups 998:1016 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 5 theme those 1070:1074 arg1 abundances 1026:1035 the abundances 1022:1035 the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins 1022:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 5 theme those 1070:1074 arg1 glycoforms 1048:1057 the IgG glycoforms 1040:1057 the IgG glycoforms as well as those of inflammatory mediating proteins 1040:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 5 theme those 1070:1074 arg1 those 1070:1074 those 1070:1074 those 1070:1074 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 8 6 theme ≤ 1139:1139 arg1 p 1137:1137 p ≤ 0.05 1137:1144 p ≤ 0.05 1137:1144 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 8 6 theme ≤ 1139:1139 arg1 differences 1124:1134 Significant differences 1112:1134 Significant differences (p ≤ 0.05) 1112:1145 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 10 7 dep similarities 1530:1541 arg1 SMCI 1558:1561 SMCI 1558:1561 SMCI 1558:1561 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 7 dep similarities 1530:1541 arg1 similarities 1530:1541 the gender similarities 1519:1541 the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI) 1519:1603 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 7 dep similarities 1530:1541 arg1 AD 1567:1568 AD 1567:1568 AD 1567:1568 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 11 8 from ability 1813:1819 arg1 females 1824:1830 females 1824:1830 females 1824:1830 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 4 9 with controls 535:542 arg1 MCI 635:637 progressive (P)MCI 620:637 progressive (P)MCI 620:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 9 with controls 535:542 arg1 AD 661:662 AD 661:662 AD 661:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 9 with controls 535:542 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 9 10 from decline 1251:1257 arg1 complexity 1272:1281 glycoform complexity 1262:1281 glycoform complexity 1262:1281 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 8 11 theme galactosylated 1193:1206 arg1 forms 1223:1227 complex galactosylated and sialylated forms 1185:1227 complex galactosylated and sialylated forms 1185:1227 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 7 12 theme glycoforms 1048:1057 arg1 groups 1011:1016 the patients groups 998:1016 the patients groups 998:1016 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 12 theme glycoforms 1048:1057 arg1 abundances 1026:1035 the abundances 1022:1035 the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins 1022:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 12 theme glycoforms 1048:1057 arg1 glycoforms 1048:1057 the IgG glycoforms 1040:1057 the IgG glycoforms as well as those of inflammatory mediating proteins 1040:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 12 theme glycoforms 1048:1057 arg1 those 1070:1074 those 1070:1074 those 1070:1074 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 10 13 theme = 1500:1500 arg1 0.36 1502:1505 Q(2)(cum) = 0.36 1490:1505 Q(2)(cum) = 0.36 1490:1505 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 9 14 theme AD 1379:1380 arg1 onset 1370:1374 the onset 1366:1374 the onset of AD 1366:1380 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 8 15 theme sialylated 1212:1221 arg1 forms 1223:1227 complex galactosylated and sialylated forms 1185:1227 complex galactosylated and sialylated forms 1185:1227 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 10 16 theme cum 1495:1497 arg1 0.36 1502:1505 Q(2)(cum) = 0.36 1490:1505 Q(2)(cum) = 0.36 1490:1505 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 2 17 theme IgG-Fc 368:373 arg1 structure 384:392 the IgG-Fc N-glycan structure 364:392 the IgG-Fc N-glycan structure 364:392 IgG functionality can drastically change from anti- to pro-inflammatory via alterations in the IgG-Fc N-glycan structure. 24028868 8 18 theme lower 1166:1170 arg1 abundance 1172:1180 a lower abundance 1164:1180 a lower abundance of complex galactosylated and sialylated forms in AD 1164:1233 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 6 19 theme glycoform 884:892 arg1 abundance 894:902 total IgGn glycoform abundance 873:902 total IgGn glycoform abundance 873:902 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 10 20 theme pro-inflammatory 1645:1660 arg1 markers 1670:1676 pro-inflammatory protein markers 1645:1676 pro-inflammatory protein markers 1645:1676 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 6 21 theme total 873:877 arg1 abundance 894:902 total IgGn glycoform abundance 873:902 total IgGn glycoform abundance 873:902 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 10 22 theme female 1683:1688 arg1 PMCI 1690:1693 female PMCI 1683:1693 female PMCI 1683:1693 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 7 23 theme patients 1002:1009 arg1 groups 1011:1016 the patients groups 998:1016 the patients groups 998:1016 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 23 theme patients 1002:1009 arg1 those 1070:1074 those 1070:1074 those 1070:1074 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 7 23 theme patients 1002:1009 arg1 glycoforms 1048:1057 the IgG glycoforms 1040:1057 the IgG glycoforms as well as those of inflammatory mediating proteins 1040:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 2 24 theme IgG 273:275 arg1 functionality 277:289 IgG functionality 273:289 IgG functionality 273:289 IgG functionality can drastically change from anti- to pro-inflammatory via alterations in the IgG-Fc N-glycan structure. 24028868 1 25 theme anti-amyloid-β 132:145 arg1 inflammation 190:201 peripheral inflammation 179:201 peripheral inflammation 179:201 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 25 theme anti-amyloid-β 132:145 arg1 factors 207:213 factors 207:213 factors correlating with development of Alzheimer's disease (AD) 207:270 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 25 theme anti-amyloid-β 132:145 arg1 IgGs 169:172 IgGs 169:172 IgGs 169:172 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 25 theme anti-amyloid-β 132:145 arg1 immunoglobulins 152:166 Blood-based anti-amyloid-β (Aβ) immunoglobulins 120:166 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) 120:173 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 10 26 theme gender 1523:1528 arg1 SMCI 1558:1561 SMCI 1558:1561 SMCI 1558:1561 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 26 theme gender 1523:1528 arg1 similarities 1530:1541 the gender similarities 1519:1541 the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI) 1519:1603 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 26 theme gender 1523:1528 arg1 AD 1567:1568 AD 1567:1568 AD 1567:1568 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 27 theme truncated 1700:1708 arg1 glycans 1717:1723 truncated IgG-Fc glycans 1700:1723 truncated IgG-Fc glycans 1700:1723 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 11 28 theme exacerbated 1887:1897 arg1 progression 1907:1917 exacerbated disease progression 1887:1917 exacerbated disease progression 1887:1917 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 5 29 theme shotgun 694:700 arg1 proteomics 702:711 Label-free shotgun proteomics 683:711 Label-free shotgun proteomics 683:711 Label-free shotgun proteomics was applied without glycoprotein enrichment. 24028868 10 30 theme Principal 1383:1391 arg1 PCA 1413:1415 PCA 1413:1415 PCA 1413:1415 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 30 theme Principal 1383:1391 arg1 analysis 1403:1410 Principal component analysis 1383:1410 Principal component analysis 1383:1410 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 2 31 theme N-glycan 375:382 arg1 structure 384:392 the IgG-Fc N-glycan structure 364:392 the IgG-Fc N-glycan structure 364:392 IgG functionality can drastically change from anti- to pro-inflammatory via alterations in the IgG-Fc N-glycan structure. 24028868 0 32 theme Blood 0:4 arg1 glycans 20:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 9 33 theme glycoform 1262:1270 arg1 complexity 1272:1281 glycoform complexity 1262:1281 glycoform complexity 1262:1281 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 4 34 theme controls 535:542 arg1 Samples 495:501 Samples 495:501 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD 495:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 2 35 dep pro-inflammatory 328:343 arg1 to 325:326 to 325:326 to 325:326 IgG functionality can drastically change from anti- to pro-inflammatory via alterations in the IgG-Fc N-glycan structure. 24028868 0 36 theme IgG 13:15 arg1 glycans 20:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 9 37 theme disease 1299:1305 arg1 progress 1307:1314 disease progress 1299:1314 disease progress 1299:1314 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 4 38 theme age-matched 508:518 arg1 subjects 520:527 age-matched subjects 508:527 age-matched subjects 508:527 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 10 39 dep Males 1418:1422 arg1 Females 1462:1468 Females 1462:1468 Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62 1418:1487 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 39 dep Males 1418:1422 arg1 0.62 1484:1487 R(2)X(cum) = 0.62 1471:1487 Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62 1418:1487 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 39 dep Males 1418:1422 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 6 40 theme EEQFNSTFR 799:807 arg1 IgG2 810:813 EEQFNSTFR (IgG2) 799:814 EEQFNSTFR (IgG2) 799:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 40 theme EEQFNSTFR 799:807 arg1 peptides 769:776 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 10 41 theme R 1425:1425 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 4 42 with patients 606:613 arg1 MCI 635:637 progressive (P)MCI 620:637 progressive (P)MCI 620:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 42 with patients 606:613 arg1 AD 661:662 AD 661:662 AD 661:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 42 with patients 606:613 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 9 43 theme inverse 1323:1329 arg1 change 1331:1336 an inverse change 1320:1336 an inverse change 1320:1336 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 10 44 theme cum 1431:1433 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 45 theme Q 1490:1490 arg1 0.36 1502:1505 Q(2)(cum) = 0.36 1490:1505 Q(2)(cum) = 0.36 1490:1505 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 7 46 theme mediating 1092:1100 arg1 proteins 1102:1109 inflammatory mediating proteins 1079:1109 inflammatory mediating proteins 1079:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 4 47 dep progressive 620:630 arg1 P 633:633 P 633:633 P 633:633 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 10 48 theme X 1429:1429 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 3 49 theme AD 491:492 arg1 development 476:486 the development 472:486 the development of AD 472:492 Herein, we tested if IgG-Fc glycosylation in plasma is indeed altered during the development of AD. 24028868 4 50 with patients 647:654 arg1 MCI 635:637 progressive (P)MCI 620:637 progressive (P)MCI 620:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 50 with patients 647:654 arg1 AD 661:662 AD 661:662 AD 661:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 50 with patients 647:654 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 10 51 dep = 1454:1454 arg1 Q 1444:1444 Q(2) 1444:1447 Q(2) 1444:1447 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 51 dep = 1454:1454 arg1 0.34 1456:1459 0.34 1456:1459 0.34 1456:1459 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 6 52 theme EEQYNSTYR 778:786 arg1 peptides 769:776 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 52 theme EEQYNSTYR 778:786 arg1 IgG1 789:792 EEQYNSTYR (IgG1) 778:793 EEQYNSTYR (IgG1) 778:793 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 4 53 with patients 548:555 arg1 MCI 635:637 progressive (P)MCI 620:637 progressive (P)MCI 620:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 53 with patients 548:555 arg1 AD 661:662 AD 661:662 AD 661:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 4 53 with patients 548:555 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 9 54 theme prior 1357:1361 arg1 males 1351:1355 males 1351:1355 males prior to the onset of AD 1351:1380 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 4 55 theme mild 569:572 arg1 impairment 584:593 stable mild cognitive impairment 562:593 stable mild cognitive impairment 562:593 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 10 56 theme = 1436:1436 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 7 57 gly glycoforms 1048:1057 arg1 IgG 1044:1046 the IgG glycoforms 1040:1057 the IgG glycoforms as well as those of inflammatory mediating proteins 1040:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 2 58 from alterations 349:359 arg1 structure 384:392 the IgG-Fc N-glycan structure 364:392 the IgG-Fc N-glycan structure 364:392 IgG functionality can drastically change from anti- to pro-inflammatory via alterations in the IgG-Fc N-glycan structure. 24028868 4 59 theme progressive 620:630 arg1 MCI 635:637 progressive (P)MCI 620:637 progressive (P)MCI 620:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 7 60 theme IgG 1044:1046 arg1 glycoforms 1048:1057 the IgG glycoforms 1040:1057 the IgG glycoforms as well as those of inflammatory mediating proteins 1040:1109 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 11 61 theme AD 1789:1790 arg1 onset 1780:1784 the onset 1776:1784 the onset of AD 1776:1790 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 6 62 dep peptides 769:776 arg1 IgG2 810:813 EEQFNSTFR (IgG2) 799:814 EEQFNSTFR (IgG2) 799:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 62 dep peptides 769:776 arg1 IgG1 789:792 EEQYNSTYR (IgG1) 778:793 EEQYNSTYR (IgG1) 778:793 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 62 dep peptides 769:776 arg1 peptides 769:776 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 8 63 theme complex 1185:1191 arg1 forms 1223:1227 complex galactosylated and sialylated forms 1185:1227 complex galactosylated and sialylated forms 1185:1227 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 10 64 theme close 1619:1623 arg1 correlation 1625:1635 a close correlation 1617:1635 a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans 1617:1723 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 1 65 theme peripheral 179:188 arg1 inflammation 190:201 peripheral inflammation 179:201 peripheral inflammation 179:201 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 65 theme peripheral 179:188 arg1 immunoglobulins 152:166 Blood-based anti-amyloid-β (Aβ) immunoglobulins 120:166 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) 120:173 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 65 theme peripheral 179:188 arg1 factors 207:213 factors 207:213 factors correlating with development of Alzheimer's disease (AD) 207:270 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 10 66 theme = 1454:1454 arg1 0.65 1438:1441 R(2)X(cum) = 0.65 1425:1441 R(2)X(cum) = 0.65 1425:1441 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 4 67 theme patients 606:613 arg1 Samples 495:501 Samples 495:501 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD 495:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 10 68 theme protein 1662:1668 arg1 markers 1670:1676 pro-inflammatory protein markers 1645:1676 pro-inflammatory protein markers 1645:1676 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 6 69 theme IgGn 879:882 arg1 abundance 894:902 total IgGn glycoform abundance 873:902 total IgGn glycoform abundance 873:902 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 4 70 theme patients 647:654 arg1 Samples 495:501 Samples 495:501 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD 495:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 8 71 gly sialylated 1212:1221 arg1 forms 1223:1227 complex galactosylated and sialylated forms 1185:1227 complex galactosylated and sialylated forms 1185:1227 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 8 72 from abundance 1172:1180 arg1 AD 1232:1233 AD 1232:1233 AD 1232:1233 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 1 73 theme Blood-based 120:130 arg1 Aβ 148:149 Aβ 148:149 Aβ 148:149 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 1 73 theme Blood-based 120:130 arg1 anti-amyloid-β 132:145 Blood-based anti-amyloid-β 120:145 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) 120:173 Blood-based anti-amyloid-β (Aβ) immunoglobulins (IgGs) and peripheral inflammation are factors correlating with development of Alzheimer's disease (AD). 24028868 4 74 theme 34 603:604 arg1 patients 606:613 34 patients 603:613 34 patients with progressive (P)MCI 603:637 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 0 75 theme progressive 81:91 arg1 impairment 108:117 progressive mild cognitive impairment 81:117 progressive mild cognitive impairment 81:117 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 3 76 gly glycosylation 423:435 arg1 plasma 440:445 plasma 440:445 plasma 440:445 Herein, we tested if IgG-Fc glycosylation in plasma is indeed altered during the development of AD. 24028868 7 77 theme multivariate 920:931 arg1 statistics 933:942 Univariate and multivariate statistics 905:942 Univariate and multivariate statistics 905:942 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 11 78 theme disease 1899:1905 arg1 progression 1907:1917 exacerbated disease progression 1887:1917 exacerbated disease progression 1887:1917 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 9 79 located found 1342:1346 arg1 males 1351:1355 males 1351:1355 males prior to the onset of AD 1351:1380 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 9 79 located found 1342:1346 arg2 change 1331:1336 an inverse change 1320:1336 an inverse change 1320:1336 For females, a decline in glycoform complexity correlated with disease progress but an inverse change was found in males prior to the onset of AD. 24028868 0 80 theme cognitive 98:106 arg1 impairment 108:117 progressive mild cognitive impairment 81:117 progressive mild cognitive impairment 81:117 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 7 81 theme Univariate 905:914 arg1 statistics 933:942 Univariate and multivariate statistics 905:942 Univariate and multivariate statistics 905:942 Univariate and multivariate statistics were employed to investigate the correlations between the patients groups and the abundances of the IgG glycoforms as well as those of inflammatory mediating proteins. 24028868 10 82 theme component 1393:1401 arg1 PCA 1413:1415 PCA 1413:1415 PCA 1413:1415 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 82 theme component 1393:1401 arg1 analysis 1403:1410 Principal component analysis 1383:1410 Principal component analysis 1383:1410 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 83 dep PCA 1413:1415 arg1 Males 1418:1422 Males 1418:1422 Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62 1418:1487 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 10 83 dep PCA 1413:1415 arg1 0.36 1502:1505 Q(2)(cum) = 0.36 1490:1505 Q(2)(cum) = 0.36 1490:1505 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 5 84 theme Label-free 683:692 arg1 proteomics 702:711 Label-free shotgun proteomics 683:711 Label-free shotgun proteomics 683:711 Label-free shotgun proteomics was applied without glycoprotein enrichment. 24028868 0 85 theme plasma 6:11 arg1 glycans 20:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 3 86 theme IgG-Fc 416:421 arg1 glycosylation 423:435 IgG-Fc glycosylation 416:435 IgG-Fc glycosylation in plasma 416:445 Herein, we tested if IgG-Fc glycosylation in plasma is indeed altered during the development of AD. 24028868 0 87 theme Fc 17:18 arg1 glycans 20:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans 0:26 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 11 88 theme lower 1807:1811 arg1 ability 1813:1819 a lower ability 1805:1819 a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression 1805:1917 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 4 89 theme patients 548:555 arg1 Samples 495:501 Samples 495:501 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD 495:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 5 90 theme glycoprotein 733:744 arg1 enrichment 746:755 glycoprotein enrichment 733:755 glycoprotein enrichment 733:755 Label-free shotgun proteomics was applied without glycoprotein enrichment. 24028868 6 91 from Glycans 758:764 arg1 peptides 769:776 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) 769:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 91 from Glycans 758:764 arg1 IgG1 789:792 EEQYNSTYR (IgG1) 778:793 EEQYNSTYR (IgG1) 778:793 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 6 91 from Glycans 758:764 arg1 IgG2 810:813 EEQFNSTFR (IgG2) 799:814 EEQFNSTFR (IgG2) 799:814 Glycans on peptides EEQYNSTYR (IgG1) and EEQFNSTFR (IgG2) were quantified, and their abundances were normalized to total IgGn glycoform abundance. 24028868 11 92 theme peripheral 1844:1853 arg1 inflammation 1855:1866 peripheral inflammation 1844:1866 peripheral inflammation 1844:1866 The differences observed between genders prior to the onset of AD may indicate a lower ability in females to suppress peripheral inflammation, which may lead to exacerbated disease progression. 24028868 0 93 theme mild 93:96 arg1 impairment 108:117 progressive mild cognitive impairment 81:117 progressive mild cognitive impairment 81:117 Blood plasma IgG Fc glycans are significantly altered in Alzheimer's disease and progressive mild cognitive impairment. 24028868 4 94 from subjects 520:527 arg1 Samples 495:501 Samples 495:501 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD 495:662 Samples from age-matched subjects of 23 controls, 58 patients with stable mild cognitive impairment (SMCI), 34 patients with progressive (P)MCI, and 31 patients with AD were investigated. 24028868 5 95 gly glycoprotein 733:744 arg1 glycoprotein 733:744 glycoprotein enrichment 733:755 glycoprotein enrichment 733:755 Label-free shotgun proteomics was applied without glycoprotein enrichment. 24028868 8 96 theme forms 1223:1227 arg1 abundance 1172:1180 a lower abundance 1164:1180 a lower abundance of complex galactosylated and sialylated forms in AD 1164:1233 Significant differences (p ≤ 0.05) were found, with a lower abundance of complex galactosylated and sialylated forms in AD. 24028868 10 97 theme IgG-Fc 1710:1715 arg1 glycans 1717:1723 truncated IgG-Fc glycans 1700:1723 truncated IgG-Fc glycans 1700:1723 Principal component analysis (PCA; Males: R(2)X(cum) = 0.65, Q(2)(cum) = 0.34; Females: R(2)X(cum) = 0.62, Q(2)(cum) = 0.36), confirmed the gender similarities (for controls, SMCI and AD) as well as differences (for PMCI), and showed a close correlation between pro-inflammatory protein markers, AD, female PMCI, and truncated IgG-Fc glycans. 24028868 3 98 from glycosylation 423:435 arg1 plasma 440:445 plasma 440:445 plasma 440:445 Herein, we tested if IgG-Fc glycosylation in plasma is indeed altered during the development of AD. 26990841 3 0 theme extended 478:485 arg1 version 487:493 An extended version 475:493 An extended version of NIST-developed software for analysis of "shotgun" proteomics 475:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 1 1 theme IgG1 242:245 arg1 material 257:264 an IgG1 reference material 239:264 an IgG1 reference material 239:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 3 2 from masses 617:622 arg1 glycopeptides 582:594 the glycopeptides 578:594 the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages 578:686 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 1 3 theme material 257:264 arg1 digestion 226:234 the tryptic digestion 214:234 the tryptic digestion of an IgG1 reference material 214:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 4 4 theme all 797:799 arg1 glycans 801:807 almost all glycans 790:807 almost all glycans reported in this study 790:830 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 3 5 theme software 513:520 arg1 version 487:493 An extended version 475:493 An extended version of NIST-developed software for analysis of "shotgun" proteomics 475:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 3 6 with peptides 645:652 arg1 cleavages 678:686 up to three missed cleavages 659:686 up to three missed cleavages 659:686 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 3 7 gly glycopeptides 582:594 arg2 glycopeptides 582:594 the glycopeptides 578:594 the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages 578:686 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 3 7 gly glycopeptides 582:594 arg1 masses 617:622 their precursor masses 601:622 their precursor masses 601:622 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 3 7 gly glycopeptides 582:594 arg1 ions 636:639 product ions 628:639 product ions 628:639 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 2 8 theme ESI 298:300 arg1 LC-MS/MS 302:309 nanospray ESI LC-MS/MS 288:309 nanospray ESI LC-MS/MS 288:309 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 8 9 from changes 1363:1369 arg1 fragmentation 1378:1390 HCD fragmentation 1374:1390 HCD fragmentation 1374:1390 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 7 10 theme glycan 1317:1322 arg1 size 1324:1327 glycan size 1317:1327 glycan size 1317:1327 Systematic variations in retention with glycan size were also noted. 26990841 8 11 gly glycopeptides 1506:1518 arg2 glycopeptides 1506:1518 glycopeptides 1506:1518 glycopeptides derived from IgG1 drugs 1506:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 1 12 theme glycopeptides 188:200 arg1 analysis 176:183 a detailed analysis 165:183 a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material 165:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 6 13 theme first 1167:1171 arg1 dimension 1183:1191 the first (high-pH) dimension 1163:1191 the first (high-pH) dimension 1163:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 14 theme glycosylated 1236:1247 arg1 peptides 1249:1256 glycosylated peptides 1236:1256 glycosylated peptides 1236:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 15 theme asialylated 1140:1150 arg1 species 1152:1158 asialylated species 1140:1158 asialylated species 1140:1158 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 4 16 theme missed 713:718 arg1 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 4 16 theme missed 713:718 arg1 cleavage 720:727 a single missed cleavage 704:727 a single missed cleavage 704:727 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 8 17 theme glycan 1415:1420 arg1 structures 1422:1431 the proposed glycan structures 1402:1431 the proposed glycan structures 1402:1431 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 6 18 theme sialylated 1122:1131 arg1 ability 1201:1207 the ability 1197:1207 the ability to inject large amounts of glycosylated peptides in the 2D studies 1197:1274 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 18 theme sialylated 1122:1131 arg1 separation 1108:1117 the improved separation 1095:1117 the improved separation of sialylated versus asialylated species in the first (high-pH) dimension 1095:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 8 19 attach derived 1520:1526 arg2 glycopeptides 1506:1518 glycopeptides 1506:1518 glycopeptides derived from IgG1 drugs 1506:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 8 19 attach derived 1520:1526 arg1 drugs 1538:1542 IgG1 drugs 1533:1542 IgG1 drugs 1533:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 5 20 theme 2D 837:838 arg1 studies 840:846 The 2D studies 833:846 The 2D studies 833:846 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 2 21 theme single 460:465 arg1 digest 467:472 a single digest 458:472 a single digest 458:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 0 22 theme Tryptic 80:86 arg1 Digest 88:93 the Tryptic Digest 76:93 the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS 76:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 8 23 theme peak-annotated 1446:1459 arg1 library 1475:1481 a peak-annotated mass spectral library 1444:1481 a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs 1444:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 2 24 theme 1D 378:379 arg1 separation 381:390 conventional 1D separation 365:390 conventional 1D separation for various digestion conditions 365:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 0 25 theme Monoclonal 100:109 arg1 Antibody 111:118 a Monoclonal Antibody 98:118 a Monoclonal Antibody Using 1D and 2D LC-MS/MS 98:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 5 26 theme masses 915:920 arg1 total 856:860 a total 854:860 a total of 247 glycopeptide ions 854:885 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 5 26 theme masses 915:920 arg1 glycans 936:942 30 glycans 933:942 30 glycans found in the 1D studies 933:966 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 5 26 theme masses 915:920 arg1 glycans 894:900 60 glycans 891:900 60 glycans 891:900 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 2 27 theme 20 431:432 arg1 fraction 434:441 fraction 434:441 fraction 434:441 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 8 28 theme spectral 1466:1473 arg1 library 1475:1481 a peak-annotated mass spectral library 1444:1481 a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs 1444:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 0 29 theme In-Depth 0:7 arg1 Characterization 9:24 In-Depth Characterization 0:24 In-Depth Characterization 0:24 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 5 30 theme ions 882:885 arg1 total 856:860 a total 854:860 a total of 247 glycopeptide ions 854:885 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 5 30 theme ions 882:885 arg1 glycans 936:942 30 glycans 933:942 30 glycans found in the 1D studies 933:966 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 5 30 theme ions 882:885 arg1 glycans 894:900 60 glycans 891:900 60 glycans 891:900 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 7 31 from variations 1288:1297 arg1 retention 1302:1310 retention 1302:1310 retention with glycan size 1302:1327 Systematic variations in retention with glycan size were also noted. 26990841 6 32 theme larger 988:993 arg1 number 995:1000 This significantly larger number 969:1000 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins 969:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 33 dep glycans 1005:1011 arg1 found 1018:1022 found 1018:1022 found in any other glycoanalysis of therapeutic glycoproteins 1018:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 2 34 theme digestion 404:412 arg1 conditions 414:423 various digestion conditions 396:423 various digestion conditions 396:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 3 35 dep three 665:669 arg1 to 662:663 to 662:663 to 662:663 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 3 36 theme product 628:634 arg1 ions 636:639 product ions 628:639 product ions 628:639 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 0 37 theme Library 39:45 arg1 Building 47:54 Spectral Library Building 30:54 Spectral Library Building 30:54 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 2 38 theme 2D-LC 443:447 arg1 study 449:453 a 20 fraction 2D-LC study 429:453 a 20 fraction 2D-LC study of a single digest 429:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 5 39 theme 1D 957:958 arg1 studies 960:966 the 1D studies 953:966 the 1D studies 953:966 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 2 40 with energies 346:353 arg1 study 449:453 a 20 fraction 2D-LC study 429:453 a 20 fraction 2D-LC study of a single digest 429:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 2 40 with energies 346:353 arg1 separation 381:390 conventional 1D separation 365:390 conventional 1D separation for various digestion conditions 365:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 6 41 theme other 1031:1035 arg1 glycoanalysis 1037:1049 any other glycoanalysis 1027:1049 any other glycoanalysis of therapeutic glycoproteins 1027:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 8 42 theme IgG1 1533:1536 arg1 drugs 1538:1542 IgG1 drugs 1533:1542 IgG1 drugs 1533:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 3 43 from ions 636:639 arg1 glycopeptides 582:594 the glycopeptides 578:594 the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages 578:686 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 2 44 theme energies 346:353 arg1 range 323:327 a wide range 316:327 a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest 316:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 3 45 theme missed 671:676 arg1 cleavages 678:686 up to three missed cleavages 659:686 up to three missed cleavages 659:686 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 2 46 theme HCD 332:334 arg1 energies 346:353 HCD collision energies 332:353 HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest 332:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 7 47 with retention 1302:1310 arg1 size 1324:1327 glycan size 1317:1327 glycan size 1317:1327 Systematic variations in retention with glycan size were also noted. 26990841 1 48 theme reference 247:255 arg1 material 257:264 an IgG1 reference material 239:264 an IgG1 reference material 239:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 6 49 theme improved 1099:1106 arg1 separation 1108:1117 the improved separation 1095:1117 the improved separation of sialylated versus asialylated species in the first (high-pH) dimension 1095:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 1 50 gly glycopeptides 188:200 arg2 glycopeptides 188:200 glycopeptides 188:200 glycopeptides produced in the tryptic digestion of an IgG1 reference material 188:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 3 51 theme NIST-developed 498:511 arg1 software 513:520 NIST-developed software 498:520 NIST-developed software for analysis of "shotgun" proteomics 498:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 6 52 theme high-pH 1174:1180 arg1 dimension 1183:1191 the first (high-pH) dimension 1163:1191 the first (high-pH) dimension 1163:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 3 53 theme shotgun 539:545 arg1 proteomics 548:557 "shotgun" proteomics 538:557 "shotgun" proteomics 538:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 1 54 theme detailed 167:174 arg1 analysis 176:183 a detailed analysis 165:183 a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material 165:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 4 55 theme glycans 801:807 arg1 determination 773:785 the determination 769:785 the determination of almost all glycans reported in this study 769:830 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 6 56 gly glycoproteins 1066:1078 arg1 glycoproteins 1066:1078 therapeutic glycoproteins 1054:1078 therapeutic glycoproteins 1054:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 2 57 theme nanospray 288:296 arg1 LC-MS/MS 302:309 nanospray ESI LC-MS/MS 288:309 nanospray ESI LC-MS/MS 288:309 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 3 58 theme " 546:546 arg1 proteomics 548:557 "shotgun" proteomics 538:557 "shotgun" proteomics 538:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 0 59 from Digest 88:93 arg1 Building 47:54 Spectral Library Building 30:54 Spectral Library Building 30:54 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 0 59 from Digest 88:93 arg1 Characterization 9:24 In-Depth Characterization 0:24 In-Depth Characterization 0:24 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 6 60 theme glycoproteins 1066:1078 arg1 glycoanalysis 1037:1049 any other glycoanalysis 1027:1049 any other glycoanalysis of therapeutic glycoproteins 1027:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 61 theme peptides 1249:1256 arg1 amounts 1225:1231 large amounts 1219:1231 large amounts of glycosylated peptides 1219:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 61 theme peptides 1249:1256 arg1 peptides 1249:1256 glycosylated peptides 1236:1256 glycosylated peptides 1236:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 62 gly glycosylated 1236:1247 arg1 peptides 1249:1256 glycosylated peptides 1236:1256 glycosylated peptides 1236:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 3 63 theme proteomics 548:557 arg1 analysis 526:533 analysis 526:533 analysis of "shotgun" proteomics 526:557 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 8 64 theme proposed 1406:1413 arg1 structures 1422:1431 the proposed glycan structures 1402:1431 the proposed glycan structures 1402:1431 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 6 65 theme species 1152:1158 arg1 ability 1201:1207 the ability 1197:1207 the ability to inject large amounts of glycosylated peptides in the 2D studies 1197:1274 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 65 theme species 1152:1158 arg1 separation 1108:1117 the improved separation 1095:1117 the improved separation of sialylated versus asialylated species in the first (high-pH) dimension 1095:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 66 theme large 1219:1223 arg1 amounts 1225:1231 large amounts 1219:1231 large amounts of glycosylated peptides 1219:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 6 66 theme large 1219:1223 arg1 peptides 1249:1256 glycosylated peptides 1236:1256 glycosylated peptides 1236:1256 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 7 67 theme Systematic 1277:1286 arg1 variations 1288:1297 Systematic variations 1277:1297 Systematic variations in retention with glycan size 1277:1327 Systematic variations in retention with glycan size were also noted. 26990841 2 68 theme digest 467:472 arg1 study 449:453 a 20 fraction 2D-LC study 429:453 a 20 fraction 2D-LC study of a single digest 429:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 2 68 theme digest 467:472 arg1 separation 381:390 conventional 1D separation 365:390 conventional 1D separation for various digestion conditions 365:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 0 69 dep 1D 126:127 arg1 LC-MS/MS 136:143 LC-MS/MS 136:143 LC-MS/MS 136:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 8 70 theme mass 1461:1464 arg1 library 1475:1481 a peak-annotated mass spectral library 1444:1481 a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs 1444:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 0 71 from Characterization 9:24 arg1 Digest 88:93 the Tryptic Digest 76:93 the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS 76:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 8 72 theme HCD 1374:1376 arg1 fragmentation 1378:1390 HCD fragmentation 1374:1390 HCD fragmentation 1374:1390 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 5 73 gly glycopeptide 869:880 arg2 glycopeptide 869:880 247 glycopeptide ions 865:885 247 glycopeptide ions 865:885 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 0 74 from Glycopeptides 59:71 arg1 Digest 88:93 the Tryptic Digest 76:93 the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS 76:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 0 75 theme Antibody 111:118 arg1 Digest 88:93 the Tryptic Digest 76:93 the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS 76:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 5 76 theme different 905:913 arg1 masses 915:920 different masses 905:920 different masses 905:920 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 6 77 theme glycans 1005:1011 arg1 number 995:1000 This significantly larger number 969:1000 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins 969:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 0 78 from Building 47:54 arg1 Digest 88:93 the Tryptic Digest 76:93 the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS 76:143 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 1 79 theme tryptic 218:224 arg1 digestion 226:234 the tryptic digestion 214:234 the tryptic digestion of an IgG1 reference material 214:264 This work presents a detailed analysis of glycopeptides produced in the tryptic digestion of an IgG1 reference material. 26990841 6 80 from separation 1108:1117 arg1 dimension 1183:1191 the first (high-pH) dimension 1163:1191 the first (high-pH) dimension 1163:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 2 81 theme conventional 365:376 arg1 separation 381:390 conventional 1D separation 365:390 conventional 1D separation for various digestion conditions 365:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 5 82 theme glycopeptide 869:880 arg1 ions 882:885 247 glycopeptide ions 865:885 247 glycopeptide ions 865:885 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 0 83 theme Spectral 30:37 arg1 Building 47:54 Spectral Library Building 30:54 Spectral Library Building 30:54 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 4 84 theme single 706:711 arg1 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 4 84 theme single 706:711 arg1 cleavage 720:727 a single missed cleavage 704:727 a single missed cleavage 704:727 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 2 85 theme various 396:402 arg1 conditions 414:423 various digestion conditions 396:423 various digestion conditions 396:423 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 8 86 theme glycopeptides 1506:1518 arg1 analysis 1494:1501 the analysis 1490:1501 the analysis of glycopeptides derived from IgG1 drugs 1490:1542 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 3 87 theme precursor 607:615 arg1 masses 617:622 their precursor masses 601:622 their precursor masses 601:622 An extended version of NIST-developed software for analysis of "shotgun" proteomics served to identify the glycopeptides from their precursor masses and product ions for peptides with up to three missed cleavages. 26990841 6 88 theme 2D 1265:1266 arg1 studies 1268:1274 the 2D studies 1261:1274 the 2D studies 1261:1274 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 0 89 theme Glycopeptides 59:71 arg1 Building 47:54 Spectral Library Building 30:54 Spectral Library Building 30:54 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 0 89 theme Glycopeptides 59:71 arg1 Characterization 9:24 In-Depth Characterization 0:24 In-Depth Characterization 0:24 In-Depth Characterization and Spectral Library Building of Glycopeptides in the Tryptic Digest of a Monoclonal Antibody Using 1D and 2D LC-MS/MS. 26990841 6 90 theme therapeutic 1054:1064 arg1 glycoproteins 1066:1078 therapeutic glycoproteins 1054:1078 therapeutic glycoproteins 1054:1078 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 2 91 theme fraction 434:441 arg1 study 449:453 a 20 fraction 2D-LC study 429:453 a 20 fraction 2D-LC study of a single digest 429:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 2 92 theme wide 318:321 arg1 range 323:327 a wide range 316:327 a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest 316:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 5 93 located found 944:948 arg1 studies 960:966 the 1D studies 953:966 the 1D studies 953:966 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 5 93 located found 944:948 arg2 glycans 936:942 30 glycans 933:942 30 glycans found in the 1D studies 933:966 The 2D studies found a total of 247 glycopeptide ions and 60 glycans of different masses, including 30 glycans found in the 1D studies. 26990841 6 94 gly asialylated 1140:1150 arg1 species 1152:1158 asialylated species 1140:1158 asialylated species 1140:1158 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 26990841 4 95 with peptide 691:697 arg1 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 TKPREEQYNSTYR 730:742 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 4 95 with peptide 691:697 arg1 cleavage 720:727 a single missed cleavage 704:727 a single missed cleavage 704:727 A peptide with a single missed cleavage, TKPREEQYNSTYR, was dominant and led to the determination of almost all glycans reported in this study. 26990841 8 96 theme Energy-dependent 1346:1361 arg1 changes 1363:1369 Energy-dependent changes 1346:1369 Energy-dependent changes in HCD fragmentation 1346:1390 Energy-dependent changes in HCD fragmentation confirmed the proposed glycan structures and led to a peak-annotated mass spectral library to aid the analysis of glycopeptides derived from IgG1 drugs. 26990841 2 97 theme collision 336:344 arg1 energies 346:353 HCD collision energies 332:353 HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest 332:472 Analysis was done by nanospray ESI LC-MS/MS over a wide range of HCD collision energies with both conventional 1D separation for various digestion conditions and a 20 fraction 2D-LC study of a single digest. 26990841 6 98 from ability 1201:1207 arg1 dimension 1183:1191 the first (high-pH) dimension 1163:1191 the first (high-pH) dimension 1163:1191 This significantly larger number of glycans than found in any other glycoanalysis of therapeutic glycoproteins is due to both the improved separation of sialylated versus asialylated species in the first (high-pH) dimension and the ability to inject large amounts of glycosylated peptides in the 2D studies. 27254475 2 0 link N-linked 315:322 arg1 glycosylation 324:336 N-linked glycosylation 315:336 N-linked glycosylation 315:336 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 6 1 theme feeding 1093:1099 arg1 approaches 1101:1110 The different feeding approaches 1079:1110 The different feeding approaches 1079:1110 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 4 2 theme ammonia 721:727 arg1 effects 698:704 the effects 694:704 the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation 694:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 5 3 theme culture 921:927 arg1 conditions 929:938 culture conditions 921:938 culture conditions as well as multiple media supplementations along the culture duration 921:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 5 4 theme conditions 929:938 arg1 shift 912:916 a single shift 903:916 a single shift of culture conditions as well as multiple media supplementations along the culture duration 903:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 5 5 theme media 960:964 arg1 supplementations 966:981 multiple media supplementations 951:981 culture conditions as well as multiple media supplementations along the culture duration 921:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 7 6 theme pattern 1263:1269 arg1 evolution 1235:1243 the time evolution 1226:1243 the time evolution of the mAb glycan pattern 1226:1269 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 5 7 theme culture 993:999 arg1 duration 1001:1008 the culture duration 989:1008 the culture duration 989:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 4 8 theme supplementation 764:778 arg1 effects 698:704 the effects 694:704 the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation 694:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 4 9 theme glycosylation 619:631 arg1 changes 600:606 the time-dependent changes 581:606 the time-dependent changes of protein glycosylation 581:631 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 0 10 theme Microbioreactor 70:84 arg1 experiments 86:96 Microbioreactor experiments 70:96 Microbioreactor experiments 70:96 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 4 11 theme nucleotide 783:792 arg1 sugars 794:799 nucleotide sugars 783:799 nucleotide sugars 783:799 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 2 12 theme N-linked 315:322 arg1 glycosylation 324:336 N-linked glycosylation 315:336 N-linked glycosylation 315:336 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 7 13 theme mAb 1252:1254 arg1 pattern 1263:1269 the mAb glycan pattern 1248:1269 the mAb glycan pattern 1248:1269 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 4 14 theme galactose 730:738 arg1 effects 698:704 the effects 694:704 the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation 694:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 4 15 link N-linked 816:823 arg1 glycosylation 825:837 mAb N-linked glycosylation 812:837 mAb N-linked glycosylation 812:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 2 16 theme cell 393:396 arg1 cultures 408:415 mammalian cell fed-batch cultures 383:415 mammalian cell fed-batch cultures 383:415 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 3 17 theme modern 456:461 arg1 processes 474:482 modern industrial processes 456:482 modern industrial processes 456:482 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 4 18 theme protein 611:617 arg1 glycosylation 619:631 protein glycosylation 611:631 protein glycosylation 611:631 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 2 19 theme culture 256:262 arg1 conditions 264:273 cell culture conditions 251:273 cell culture conditions 251:273 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 2 20 theme fed-batch 398:406 arg1 cultures 408:415 mammalian cell fed-batch cultures 383:415 mammalian cell fed-batch cultures 383:415 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 0 21 link N-linked 38:45 arg1 glycosylation 47:59 mAb N-linked glycosylation 34:59 mAb N-linked glycosylation 34:59 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 4 22 theme chloride 755:762 arg1 supplementation 764:778 manganese chloride supplementation 745:778 manganese chloride supplementation 745:778 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 7 23 theme different 1322:1330 arg1 structures 1346:1355 different glycosylation structures 1322:1355 different glycosylation structures 1322:1355 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 1 24 theme biotherapeutic 168:181 arg1 proteins 183:190 many biotherapeutic proteins 163:190 many biotherapeutic proteins such as monoclonal antibodies (mAbs) 163:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 1 24 theme biotherapeutic 168:181 arg1 antibodies 211:220 monoclonal antibodies 200:220 monoclonal antibodies (mAbs) 200:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 4 25 theme manganese 745:753 arg1 supplementation 764:778 manganese chloride supplementation 745:778 manganese chloride supplementation 745:778 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 1 26 theme proteins 183:190 arg1 efficacy 151:158 the efficacy 147:158 the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs) 147:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 7 27 theme time 1230:1233 arg1 evolution 1235:1243 the time evolution 1226:1243 the time evolution of the mAb glycan pattern 1226:1269 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 6 28 theme glycosylation 1129:1141 arg1 patterns 1143:1150 constant glycosylation patterns 1120:1150 constant glycosylation patterns 1120:1150 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 2 29 gly glycoproteins 363:375 arg1 glycoproteins 363:375 glycoproteins 363:375 glycoproteins using mammalian cell fed-batch cultures 363:415 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 5 30 theme multiple 951:958 arg1 supplementations 966:981 multiple media supplementations 951:981 culture conditions as well as multiple media supplementations along the culture duration 921:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 6 31 theme different 1083:1091 arg1 approaches 1101:1110 The different feeding approaches 1079:1110 The different feeding approaches 1079:1110 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 5 32 theme constant 1046:1053 arg1 profiles 1069:1076 changing and constant glycosylation profiles 1033:1076 changing and constant glycosylation profiles 1033:1076 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 3 33 theme concentrated 489:500 arg1 additions 507:515 concentrated feed additions 489:515 concentrated feed additions 489:515 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 0 34 theme time 16:19 arg1 evolution 21:29 the time evolution 12:29 the time evolution of mAb N-linked glycosylation 12:59 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 0 34 theme time 16:19 arg1 Part 62:65 Part I 62:67 Part I 62:67 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 4 35 theme time-dependent 585:598 arg1 changes 600:606 the time-dependent changes 581:606 the time-dependent changes of protein glycosylation 581:631 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 5 36 theme supplementations 966:981 arg1 shift 912:916 a single shift 903:916 a single shift of culture conditions as well as multiple media supplementations along the culture duration 903:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 1 37 theme N-linked 99:106 arg1 glycosylation 108:120 N-linked glycosylation 99:120 N-linked glycosylation 99:120 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 3 38 theme feed 502:505 arg1 additions 507:515 concentrated feed additions 489:515 concentrated feed additions 489:515 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 0 39 theme mAb 34:36 arg1 glycosylation 47:59 mAb N-linked glycosylation 34:59 mAb N-linked glycosylation 34:59 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 4 40 from effects 698:704 arg1 sugars 794:799 nucleotide sugars 783:799 nucleotide sugars 783:799 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 4 40 from effects 698:704 arg1 glycosylation 825:837 mAb N-linked glycosylation 812:837 mAb N-linked glycosylation 812:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 5 41 theme different 868:876 arg1 strategies 878:887 Two different strategies 864:887 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration 864:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 4 42 theme N-linked 816:823 arg1 glycosylation 825:837 mAb N-linked glycosylation 812:837 mAb N-linked glycosylation 812:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 2 43 theme mammalian 383:391 arg1 cultures 408:415 mammalian cell fed-batch cultures 383:415 mammalian cell fed-batch cultures 383:415 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 1 44 theme monoclonal 200:209 arg1 mAbs 223:226 mAbs 223:226 mAbs 223:226 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 1 44 theme monoclonal 200:209 arg1 antibodies 211:220 monoclonal antibodies 200:220 monoclonal antibodies (mAbs) 200:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 4 45 theme pH 717:718 arg1 effects 698:704 the effects 694:704 the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation 694:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 5 46 theme glycosylation 1055:1067 arg1 profiles 1069:1076 changing and constant glycosylation profiles 1033:1076 changing and constant glycosylation profiles 1033:1076 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 6 47 theme constant 1120:1127 arg1 patterns 1143:1150 constant glycosylation patterns 1120:1150 constant glycosylation patterns 1120:1150 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 0 48 theme glycosylation 47:59 arg1 evolution 21:29 the time evolution 12:29 the time evolution of mAb N-linked glycosylation 12:59 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 0 48 theme glycosylation 47:59 arg1 Part 62:65 Part I 62:67 Part I 62:67 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 6 49 theme culture 1174:1180 arg1 duration 1182:1189 the entire culture duration 1163:1189 the entire culture duration 1163:1189 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 3 50 theme cell 521:524 arg1 densities 526:534 cell densities 521:534 cell densities 521:534 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 0 51 theme N-linked 38:45 arg1 glycosylation 47:59 mAb N-linked glycosylation 34:59 mAb N-linked glycosylation 34:59 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 7 52 theme glycan 1256:1261 arg1 pattern 1263:1269 the mAb glycan pattern 1248:1269 the mAb glycan pattern 1248:1269 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 4 53 theme time-dependent 844:857 arg1 way 859:861 a time-dependent way 842:861 a time-dependent way 842:861 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 7 54 theme glycosylation 1332:1344 arg1 structures 1346:1355 different glycosylation structures 1322:1355 different glycosylation structures 1322:1355 By modulating the time evolution of the mAb glycan pattern, not only the endpoint but also the ratios between different glycosylation structures could be modified. 27254475 6 55 theme different 1194:1202 arg1 levels 1204:1209 different levels 1194:1209 different levels 1194:1209 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 2 56 theme Media 230:234 arg1 components 236:245 Media components 230:245 Media components 230:245 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 4 57 theme mAb 812:814 arg1 glycosylation 825:837 mAb N-linked glycosylation 812:837 mAb N-linked glycosylation 812:837 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 2 58 theme cell 251:254 arg1 conditions 264:273 cell culture conditions 251:273 cell culture conditions 251:273 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 4 59 theme culture 709:715 arg1 pH 717:718 culture pH 709:718 culture pH 709:718 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 5 60 theme single 905:910 arg1 shift 912:916 a single shift 903:916 a single shift of culture conditions as well as multiple media supplementations along the culture duration 903:1008 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 8 61 theme Chemical 1405:1412 arg1 Engineers 1414:1422 Chemical Engineers 1405:1422 Chemical Engineers 1405:1422 © 2016 American Institute of Chemical Engineers Biotechnol. 27254475 4 62 used used 674:677 arg2 system 663:668 an automated microbioreactor system 634:668 an automated microbioreactor system 634:668 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 1 63 theme key 128:130 arg1 importance 132:141 key importance 128:141 key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs) 128:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 3 64 theme industrial 463:472 arg1 processes 474:482 modern industrial processes 456:482 modern industrial processes 456:482 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 6 65 theme entire 1167:1172 arg1 duration 1182:1189 the entire culture duration 1163:1189 the entire culture duration 1163:1189 The different feeding approaches enabled constant glycosylation patterns throughout the entire culture duration at different levels. 27254475 5 66 theme changing 1033:1040 arg1 profiles 1069:1076 changing and constant glycosylation profiles 1033:1076 changing and constant glycosylation profiles 1033:1076 Two different strategies comprising of a single shift of culture conditions as well as multiple media supplementations along the culture duration were applied to obtain changing and constant glycosylation profiles. 27254475 8 67 dep Biotechnol 1424:1433 arg1 Institute 1392:1400 Institute 1392:1400 Institute 1392:1400 © 2016 American Institute of Chemical Engineers Biotechnol. 27254475 3 68 theme × 545:545 arg1 cells/mL 551:558 2 × 107 cells/mL 543:558 2 × 107 cells/mL 543:558 These parameters inevitably change in modern industrial processes with concentrated feed additions and cell densities beyond 2 × 107 cells/mL. 27254475 1 69 theme many 163:166 arg1 proteins 183:190 many biotherapeutic proteins 163:190 many biotherapeutic proteins such as monoclonal antibodies (mAbs) 163:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 1 69 theme many 163:166 arg1 antibodies 211:220 monoclonal antibodies 200:220 monoclonal antibodies (mAbs) 200:227 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 4 70 theme microbioreactor 647:661 arg1 system 663:668 an automated microbioreactor system 634:668 an automated microbioreactor system 634:668 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 27254475 1 71 link N-linked 99:106 arg1 glycosylation 108:120 N-linked glycosylation 99:120 N-linked glycosylation 99:120 N-linked glycosylation is of key importance for the efficacy of many biotherapeutic proteins such as monoclonal antibodies (mAbs). 27254475 0 72 dep Controlling 0:10 arg1 experiments 86:96 Microbioreactor experiments 70:96 Microbioreactor experiments 70:96 Controlling the time evolution of mAb N-linked glycosylation, Part I: Microbioreactor experiments. 27254475 2 73 theme glycoproteins 363:375 arg1 production 349:358 the production 345:358 the production of glycoproteins using mammalian cell fed-batch cultures 345:415 Media components and cell culture conditions have been shown to significantly affect N-linked glycosylation during the production of glycoproteins using mammalian cell fed-batch cultures. 27254475 4 74 theme automated 637:645 arg1 system 663:668 an automated microbioreactor system 634:668 an automated microbioreactor system 634:668 In order to control the time-dependent changes of protein glycosylation, an automated microbioreactor system was used to investigate the effects of culture pH, ammonia, galactose, and manganese chloride supplementation on nucleotide sugars as well as mAb N-linked glycosylation in a time-dependent way. 25251695 0 0 theme STRUBBELIG 96:105 arg1 domain 74:79 the misfolded extracellular domain 46:79 the misfolded extracellular domain of Arabidopsis STRUBBELIG 46:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 0 0 theme STRUBBELIG 96:105 arg1 STRUBBELIG 96:105 Arabidopsis STRUBBELIG 84:105 Arabidopsis STRUBBELIG 84:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 1 1 from determination 215:227 arg1 ER 306:307 ER 306:307 ER 306:307 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 1 1 from determination 215:227 arg1 reticulum 295:303 the endoplasmic reticulum 279:303 the endoplasmic reticulum (ER) 279:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 7 2 theme N-glycans 1298:1306 arg1 C-branch 1286:1293 the C-branch 1282:1293 the C-branch of N-glycans 1282:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 9 3 theme Glc1Man7GlcNAc2 1491:1505 arg1 N-glycans 1507:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 0 4 theme Arabidopsis 84:94 arg1 STRUBBELIG 96:105 Arabidopsis STRUBBELIG 84:105 Arabidopsis STRUBBELIG 84:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 4 5 theme SUB 741:743 arg1 domain 760:765 the STRUBBELIG (SUB) extracellular domain 725:765 the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) 725:778 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 4 5 theme SUB 741:743 arg1 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 6 6 theme SUBEX-C57Y 1057:1066 arg1 degradation 1068:1078 SUBEX-C57Y degradation 1057:1078 SUBEX-C57Y degradation 1057:1078 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 0 7 theme endoplasmic-reticulum-associated 110:141 arg1 degradation 143:153 endoplasmic-reticulum-associated degradation 110:153 endoplasmic-reticulum-associated degradation 110:153 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 2 8 gly glycoproteins 474:486 arg1 glycoproteins 474:486 improperly-folded glycoproteins 456:486 improperly-folded glycoproteins 456:486 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 1 9 gly glycoproteins 262:274 arg1 glycoproteins 262:274 newly synthesized glycoproteins 244:274 newly synthesized glycoproteins 244:274 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 4 10 theme glycan-dependent 797:812 arg1 manner 814:819 a glycan-dependent manner 795:819 a glycan-dependent manner in plants 795:829 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 3 11 dep mechanisms 552:561 arg1 the 548:550 the 548:550 the 548:550 In plants, the mechanisms and factors that recognize non-native proteins and sort them to ERAD are poorly understood. 25251695 9 12 theme futile 1701:1706 arg1 folding 1708:1714 futile folding attempts 1701:1723 futile folding attempts 1701:1723 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 4 13 from manner 814:819 arg1 plants 824:829 plants 824:829 plants 824:829 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 2 14 theme oligosaccharide 320:334 arg1 structures 336:345 Specific oligosaccharide structures 311:345 Specific oligosaccharide structures 311:345 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 5 15 theme α-mannosidases 968:981 arg1 inhibitor 955:963 a potent inhibitor 946:963 a potent inhibitor of α-mannosidases 946:981 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 5 15 theme α-mannosidases 968:981 arg1 kifunensine 933:943 kifunensine 933:943 kifunensine 933:943 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 4 16 theme domain 760:765 arg1 variant 714:720 a misfolded variant 702:720 a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) 702:778 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 7 17 link α1,6-linked 1259:1269 arg1 mannose 1271:1277 a terminal α1,6-linked mannose 1248:1277 a terminal α1,6-linked mannose on the C-branch of N-glycans 1248:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 2 18 theme Specific 311:318 arg1 structures 336:345 Specific oligosaccharide structures 311:345 Specific oligosaccharide structures 311:345 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 9 19 theme control 1548:1554 arg1 interconnected 1629:1642 interconnected 1629:1642 interconnected 1629:1642 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 9 19 theme control 1548:1554 arg1 processes 1556:1564 the ER-quality control processes 1533:1564 the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD 1533:1615 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 2 20 theme improperly-folded 456:472 arg1 glycoproteins 474:486 improperly-folded glycoproteins 456:486 improperly-folded glycoproteins 456:486 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 7 21 theme terminal 1250:1257 arg1 mannose 1271:1277 a terminal α1,6-linked mannose 1248:1277 a terminal α1,6-linked mannose on the C-branch of N-glycans 1248:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 9 22 theme considerable 1467:1478 arg1 amounts 1480:1486 considerable amounts 1467:1486 considerable amounts of Glc1Man7GlcNAc2 N-glycans 1467:1515 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 9 22 theme considerable 1467:1478 arg1 N-glycans 1507:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 4 23 theme misfolded 704:712 arg1 variant 714:720 a misfolded variant 702:720 a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) 702:778 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 0 24 theme context-independent 2:20 arg1 signal 31:36 A context-independent N-glycan signal 0:36 A context-independent N-glycan signal 0:36 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 9 25 theme Structural 1413:1422 arg1 analysis 1424:1431 Structural analysis 1413:1431 Structural analysis 1413:1431 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 7 26 dep α-mannosidases 1205:1218 arg1 α-mannosidases 1205:1218 the α-mannosidases MNS4 and MNS5 1201:1232 the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans 1201:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 7 26 dep α-mannosidases 1205:1218 arg1 MNS5 1229:1232 MNS5 1229:1232 MNS5 1229:1232 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 7 26 dep α-mannosidases 1205:1218 arg1 MNS4 1220:1223 MNS4 1220:1223 MNS4 1220:1223 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 5 27 with glycoprotein 861:872 arg1 N-glycans 885:893 three N-glycans 879:893 three N-glycans 879:893 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 6 28 theme associated 1122:1131 arg1 SEL1L 1145:1149 its associated ERAD factor SEL1L 1118:1149 its associated ERAD factor SEL1L 1118:1149 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 5 29 theme ER-retained 849:859 arg1 SUBEX-C57Y 832:841 SUBEX-C57Y 832:841 SUBEX-C57Y 832:841 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 5 29 theme ER-retained 849:859 arg1 glycoprotein 861:872 an ER-retained glycoprotein 846:872 an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases 846:981 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 9 30 theme ER-quality 1537:1546 arg1 interconnected 1629:1642 interconnected 1629:1642 interconnected 1629:1642 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 9 30 theme ER-quality 1537:1546 arg1 processes 1556:1564 the ER-quality control processes 1533:1564 the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD 1533:1615 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 6 31 theme knockout 1026:1033 arg1 mutants 1035:1041 Arabidopsis thaliana knockout mutants 1005:1041 Arabidopsis thaliana knockout mutants 1005:1041 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 2 32 theme molecular 355:363 arg1 chaperones 365:374 molecular chaperones 355:374 molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD) 355:534 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 1 33 theme fate 236:239 arg1 determination 215:227 the determination 211:227 the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER) 211:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 0 34 theme N-glycan 22:29 arg1 signal 31:36 A context-independent N-glycan signal 0:36 A context-independent N-glycan signal 0:36 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 1 35 gly N-glycosylation 156:170 arg1 proteins 175:182 proteins 175:182 proteins 175:182 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 5 36 dep kifunensine 933:943 arg1 the 917:919 the 917:919 the 917:919 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 5 36 dep kifunensine 933:943 arg1 presence 921:928 presence 921:928 presence 921:928 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 9 37 dep folding 1708:1714 arg1 attempts 1716:1723 attempts 1716:1723 attempts 1716:1723 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 6 38 theme Arabidopsis 1005:1015 arg1 mutants 1035:1041 Arabidopsis thaliana knockout mutants 1005:1041 Arabidopsis thaliana knockout mutants 1005:1041 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 1 39 from reticulum 295:303 arg1 determination 215:227 the determination 211:227 the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER) 211:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 9 40 theme folding 1708:1714 arg1 termination 1686:1696 termination 1686:1696 termination of futile folding attempts 1686:1723 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 6 41 theme lectin 1103:1108 arg1 OS9 1110:1112 the ER lectin OS9 1096:1112 the ER lectin OS9 1096:1112 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 6 42 theme factor 1138:1143 arg1 SEL1L 1145:1149 its associated ERAD factor SEL1L 1118:1149 its associated ERAD factor SEL1L 1118:1149 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 9 43 theme N-glycans 1507:1515 arg1 amounts 1480:1486 considerable amounts 1467:1486 considerable amounts of Glc1Man7GlcNAc2 N-glycans 1467:1515 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 9 43 theme N-glycans 1507:1515 arg1 N-glycans 1507:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Glc1Man7GlcNAc2 N-glycans 1491:1515 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 2 44 theme glycoproteins 474:486 arg1 clearance 443:451 the clearance 439:451 the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD) 439:534 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 0 45 theme extracellular 60:72 arg1 domain 74:79 the misfolded extracellular domain 46:79 the misfolded extracellular domain of Arabidopsis STRUBBELIG 46:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 0 45 theme extracellular 60:72 arg1 STRUBBELIG 96:105 Arabidopsis STRUBBELIG 84:105 Arabidopsis STRUBBELIG 84:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 7 46 theme α1,6-linked 1259:1269 arg1 mannose 1271:1277 a terminal α1,6-linked mannose 1248:1277 a terminal α1,6-linked mannose on the C-branch of N-glycans 1248:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 6 47 theme ER 1100:1101 arg1 OS9 1110:1112 the ER lectin OS9 1096:1112 the ER lectin OS9 1096:1112 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 7 48 from mannose 1271:1277 arg1 C-branch 1286:1293 the C-branch 1282:1293 the C-branch of N-glycans 1282:1306 SUBEX-C57Y was also stabilized in plants lacking the α-mannosidases MNS4 and MNS5 that generate a terminal α1,6-linked mannose on the C-branch of N-glycans. 25251695 2 49 theme mannose-binding 400:414 arg1 lectins 416:422 mannose-binding lectins 400:422 mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD) 400:534 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 0 50 theme misfolded 50:58 arg1 domain 74:79 the misfolded extracellular domain 46:79 the misfolded extracellular domain of Arabidopsis STRUBBELIG 46:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 0 50 theme misfolded 50:58 arg1 STRUBBELIG 96:105 Arabidopsis STRUBBELIG 84:105 Arabidopsis STRUBBELIG 84:105 A context-independent N-glycan signal targets the misfolded extracellular domain of Arabidopsis STRUBBELIG to endoplasmic-reticulum-associated degradation. 25251695 6 51 theme Stable 984:989 arg1 expression 991:1000 Stable expression 984:1000 Stable expression in Arabidopsis thaliana knockout mutants 984:1041 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 8 52 theme specific 1376:1383 arg1 position 1385:1392 a specific position 1374:1392 a specific position within SUBEX-C57Y 1374:1410 Notably, the glycan signal for degradation is not constrained to a specific position within SUBEX-C57Y. 25251695 5 53 gly glycoprotein 861:872 arg1 SUBEX-C57Y 832:841 SUBEX-C57Y 832:841 SUBEX-C57Y 832:841 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 5 53 gly glycoprotein 861:872 arg1 glycoprotein 861:872 an ER-retained glycoprotein 846:872 an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases 846:981 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 1 54 from fate 236:239 arg1 ER 306:307 ER 306:307 ER 306:307 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 1 54 from fate 236:239 arg1 reticulum 295:303 the endoplasmic reticulum 279:303 the endoplasmic reticulum (ER) 279:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 4 55 theme extracellular 746:758 arg1 domain 760:765 the STRUBBELIG (SUB) extracellular domain 725:765 the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) 725:778 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 4 55 theme extracellular 746:758 arg1 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 6 56 dep Arabidopsis 1005:1015 arg1 thaliana 1017:1024 thaliana 1017:1024 thaliana 1017:1024 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 4 57 theme STRUBBELIG 729:738 arg1 domain 760:765 the STRUBBELIG (SUB) extracellular domain 725:765 the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) 725:778 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 4 57 theme STRUBBELIG 729:738 arg1 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 SUBEX-C57Y 768:777 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 1 58 theme proteins 175:182 arg1 N-glycosylation 156:170 N-glycosylation 156:170 N-glycosylation of proteins 156:182 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 1 59 theme glycoproteins 262:274 arg1 fate 236:239 the fate 232:239 the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER) 232:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 6 60 from expression 991:1000 arg1 mutants 1035:1041 Arabidopsis thaliana knockout mutants 1005:1041 Arabidopsis thaliana knockout mutants 1005:1041 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 5 61 theme potent 948:953 arg1 inhibitor 955:963 a potent inhibitor 946:963 a potent inhibitor of α-mannosidases 946:981 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 5 61 theme potent 948:953 arg1 kifunensine 933:943 kifunensine 933:943 kifunensine 933:943 SUBEX-C57Y is an ER-retained glycoprotein with three N-glycans that is stabilized in the presence of kifunensine, a potent inhibitor of α-mannosidases. 25251695 9 62 theme protein 1655:1661 arg1 folding 1663:1669 protein folding 1655:1669 protein folding 1655:1669 Structural analysis revealed that SUBEX-C57Y harbours considerable amounts of Glc1Man7GlcNAc2 N-glycans suggesting that the ER-quality control processes involving calnexin/calreticulin (CNX/CRT) and ERAD are tightly interconnected to promote protein folding or disposal by termination of futile folding attempts. 25251695 3 63 theme non-native 590:599 arg1 proteins 601:608 non-native proteins 590:608 non-native proteins 590:608 In plants, the mechanisms and factors that recognize non-native proteins and sort them to ERAD are poorly understood. 25251695 4 64 theme present 662:668 arg1 study 670:674 the present study 658:674 the present study 658:674 In the present study, we provide evidence that a misfolded variant of the STRUBBELIG (SUB) extracellular domain (SUBEX-C57Y) is degraded in a glycan-dependent manner in plants. 25251695 1 65 theme synthesized 250:260 arg1 glycoproteins 262:274 newly synthesized glycoproteins 244:274 newly synthesized glycoproteins 244:274 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 8 66 theme glycan 1322:1327 arg1 signal 1329:1334 the glycan signal 1318:1334 the glycan signal for degradation 1318:1350 Notably, the glycan signal for degradation is not constrained to a specific position within SUBEX-C57Y. 25251695 6 67 theme ERAD 1133:1136 arg1 SEL1L 1145:1149 its associated ERAD factor SEL1L 1118:1149 its associated ERAD factor SEL1L 1118:1149 Stable expression in Arabidopsis thaliana knockout mutants revealed that SUBEX-C57Y degradation is dependent on the ER lectin OS9 and its associated ERAD factor SEL1L. 25251695 2 68 theme ER-associated 503:515 arg1 ERAD 530:533 ERAD 530:533 ERAD 530:533 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 2 68 theme ER-associated 503:515 arg1 degradation 517:527 ER-associated degradation 503:527 ER-associated degradation (ERAD) 503:534 Specific oligosaccharide structures recruit molecular chaperones that promote folding or mannose-binding lectins that assist in the clearance of improperly-folded glycoproteins by delivery to ER-associated degradation (ERAD). 25251695 1 69 theme important 193:201 arg1 role 203:206 an important role 190:206 an important role 190:206 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 1 70 theme endoplasmic 283:293 arg1 ER 306:307 ER 306:307 ER 306:307 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 25251695 1 70 theme endoplasmic 283:293 arg1 reticulum 295:303 the endoplasmic reticulum 279:303 the endoplasmic reticulum (ER) 279:308 N-glycosylation of proteins plays an important role in the determination of the fate of newly synthesized glycoproteins in the endoplasmic reticulum (ER). 24533620 7 0 theme proline-rich 1062:1073 arg1 peptide 1075:1081 this semiflexible proline-rich peptide 1044:1081 this semiflexible proline-rich peptide 1044:1081 Additionally, the flexibility of this semiflexible proline-rich peptide is significantly reduced by glycosylation. 24533620 7 1 theme peptide 1075:1081 arg1 flexibility 1029:1039 the flexibility 1025:1039 the flexibility of this semiflexible proline-rich peptide 1025:1081 Additionally, the flexibility of this semiflexible proline-rich peptide is significantly reduced by glycosylation. 24533620 3 2 dep example 381:387 arg1 constructed 528:538 constructed 528:538 are constructed to probe how glycosylation affects the physical properties of the linker 524:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 1 3 theme carbohydrate 155:166 arg1 residues 168:175 carbohydrate residues 155:175 carbohydrate residues 155:175 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 3 4 theme linker 364:369 arg1 conformation 325:336 the conformation 321:336 the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker 321:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 0 5 from Effects 0:6 arg1 linker 62:67 a semiflexible peptide linker 39:67 a semiflexible peptide linker 39:67 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 3 6 theme peptide 478:484 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 7 theme semiflexible 343:354 arg1 linker 364:369 a semiflexible peptide linker 341:369 a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker 341:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 8 from example 381:387 arg1 sets 439:442 immunoglobulin A. Three sets 415:442 immunoglobulin A. Three sets of atomistic models of this hinge peptide 415:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 8 from example 381:387 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 2 9 from monomers 225:232 arg1 range 207:211 range 207:211 range 207:211 These attached carbohydrates range from single monomers to elaborate branched glycans. 24533620 1 10 theme residues 168:175 arg1 addition 143:150 the addition 139:150 the addition of carbohydrate residues 139:175 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 3 11 theme peptide 356:362 arg1 linker 364:369 a semiflexible peptide linker 341:369 a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker 341:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 4 12 theme predominant 662:672 arg1 conformations 674:686 the predominant conformations 658:686 the predominant conformations of the peptide 658:701 We found that glycosylation greatly altered the predominant conformations of the peptide, causing it to become elongated in reference to the unglycosylated form. 24533620 6 13 gly glycosylated 989:1000 arg1 serines 1002:1008 glycosylated serines 989:1008 glycosylated serines 989:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 3 14 theme peptide 402:408 arg1 example 381:387 the example 377:387 the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker 377:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 15 theme A. 430:431 arg1 sets 439:442 immunoglobulin A. Three sets 415:442 immunoglobulin A. Three sets of atomistic models of this hinge peptide 415:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 15 theme A. 430:431 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 6 16 theme glycosylated 989:1000 arg1 serines 1002:1008 glycosylated serines 989:1008 glycosylated serines 989:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 1 17 theme essential 90:98 arg1 Glycosylation 70:82 Glycosylation 70:82 Glycosylation 70:82 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 1 17 theme essential 90:98 arg1 modification 100:111 an essential modification 87:111 an essential modification of proteins and lipids by the addition of carbohydrate residues 87:175 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 6 18 located found 894:898 arg2 glycans 882:888 glycans 882:888 glycans 882:888 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 6 18 located found 894:898 arg1 level 875:879 the residue level 863:879 the residue level 863:879 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 0 19 theme O-glycosylation 20:34 arg1 Effects 0:6 Effects 0:6 Effects of branched O-glycosylation on a semiflexible peptide linker 0:67 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 4 20 theme unglycosylated 755:768 arg1 form 770:773 the unglycosylated form 751:773 the unglycosylated form 751:773 We found that glycosylation greatly altered the predominant conformations of the peptide, causing it to become elongated in reference to the unglycosylated form. 24533620 0 21 theme branched 11:18 arg1 O-glycosylation 20:34 branched O-glycosylation 11:34 branched O-glycosylation 11:34 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 6 22 theme elements 976:983 arg1 formation 928:936 the formation 924:936 the formation of more extended secondary structural elements for glycosylated serines 924:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 2 23 theme attached 184:191 arg1 carbohydrates 193:205 These attached carbohydrates 178:205 These attached carbohydrates 178:205 These attached carbohydrates range from single monomers to elaborate branched glycans. 24533620 5 24 theme conformational 817:830 arg1 exploration 832:842 the conformational exploration 813:842 the conformational exploration of the peptide 813:857 Furthermore, glycosylation restricts the conformational exploration of the peptide. 24533620 6 25 theme structural 965:974 arg1 elements 976:983 more extended secondary structural elements 941:983 more extended secondary structural elements for glycosylated serines 941:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 2 26 theme branched 247:254 arg1 glycans 256:262 branched glycans 247:262 branched glycans 247:262 These attached carbohydrates range from single monomers to elaborate branched glycans. 24533620 3 27 theme atomistic 447:455 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 28 theme immunoglobulin 415:428 arg1 sets 439:442 immunoglobulin A. Three sets 415:442 immunoglobulin A. Three sets of atomistic models of this hinge peptide 415:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 28 theme immunoglobulin 415:428 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 6 29 theme secondary 955:963 arg1 elements 976:983 more extended secondary structural elements 941:983 more extended secondary structural elements for glycosylated serines 941:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 3 30 theme linker 606:611 arg1 properties 588:597 the physical properties 575:597 the physical properties of the linker 575:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 31 theme models 457:462 arg1 sets 439:442 immunoglobulin A. Three sets 415:442 immunoglobulin A. Three sets of atomistic models of this hinge peptide 415:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 31 theme models 457:462 arg1 models 457:462 atomistic models 447:462 atomistic models of this hinge peptide 447:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 0 32 theme peptide 54:60 arg1 linker 62:67 a semiflexible peptide linker 39:67 a semiflexible peptide linker 39:67 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 6 33 theme extended 946:953 arg1 elements 976:983 more extended secondary structural elements 941:983 more extended secondary structural elements for glycosylated serines 941:1008 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 3 34 theme glycosylation 299:311 arg1 level 290:294 the level 286:294 the level of glycosylation 286:311 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 1 35 theme proteins 116:123 arg1 Glycosylation 70:82 Glycosylation 70:82 Glycosylation 70:82 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 1 35 theme proteins 116:123 arg1 modification 100:111 an essential modification 87:111 an essential modification of proteins and lipids by the addition of carbohydrate residues 87:175 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 0 36 theme semiflexible 41:52 arg1 linker 62:67 a semiflexible peptide linker 39:67 a semiflexible peptide linker 39:67 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 2 37 theme single 218:223 arg1 monomers 225:232 single monomers 218:232 single monomers 218:232 These attached carbohydrates range from single monomers to elaborate branched glycans. 24533620 0 38 gly O-glycosylation 20:34 arg1 peptide 54:60 a semiflexible peptide linker 39:67 a semiflexible peptide linker 39:67 Effects of branched O-glycosylation on a semiflexible peptide linker. 24533620 3 39 theme varying 491:497 arg1 degrees 499:505 varying degrees 491:505 varying degrees of glycosylation 491:522 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 7 40 theme semiflexible 1049:1060 arg1 peptide 1075:1081 this semiflexible proline-rich peptide 1044:1081 this semiflexible proline-rich peptide 1044:1081 Additionally, the flexibility of this semiflexible proline-rich peptide is significantly reduced by glycosylation. 24533620 4 41 theme peptide 695:701 arg1 conformations 674:686 the predominant conformations 658:686 the predominant conformations of the peptide 658:701 We found that glycosylation greatly altered the predominant conformations of the peptide, causing it to become elongated in reference to the unglycosylated form. 24533620 1 42 theme lipids 129:134 arg1 Glycosylation 70:82 Glycosylation 70:82 Glycosylation 70:82 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 1 42 theme lipids 129:134 arg1 modification 100:111 an essential modification 87:111 an essential modification of proteins and lipids by the addition of carbohydrate residues 87:175 Glycosylation is an essential modification of proteins and lipids by the addition of carbohydrate residues. 24533620 6 43 theme residue 867:873 arg1 level 875:879 the residue level 863:879 the residue level 863:879 At the residue level, glycans are found to introduce a bias for the formation of more extended secondary structural elements for glycosylated serines. 24533620 3 44 gly glycosylation 553:565 arg1 linker 606:611 the linker 602:611 the linker 602:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 45 theme hinge 472:476 arg1 peptide 478:484 this hinge peptide 467:484 this hinge peptide 467:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 4 46 gly unglycosylated 755:768 arg1 form 770:773 the unglycosylated form 751:773 the unglycosylated form 751:773 We found that glycosylation greatly altered the predominant conformations of the peptide, causing it to become elongated in reference to the unglycosylated form. 24533620 3 47 theme glycosylation 510:522 arg1 degrees 499:505 varying degrees 491:505 varying degrees of glycosylation 491:522 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 48 theme hinge 396:400 arg1 peptide 402:408 the hinge peptide 392:408 the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide 392:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 5 49 theme peptide 851:857 arg1 exploration 832:842 the conformational exploration 813:842 the conformational exploration of the peptide 813:857 Furthermore, glycosylation restricts the conformational exploration of the peptide. 24533620 3 50 from sets 439:442 arg1 peptide 402:408 the hinge peptide 392:408 the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide 392:484 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 50 from sets 439:442 arg1 example 381:387 the example 377:387 the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker 377:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 24533620 3 51 theme physical 579:586 arg1 properties 588:597 the physical properties 575:597 the physical properties of the linker 575:611 Here, we examine how the level of glycosylation affects the conformation of a semiflexible peptide linker using the example of the hinge peptide from immunoglobulin A. Three sets of atomistic models of this hinge peptide with varying degrees of glycosylation are constructed to probe how glycosylation affects the physical properties of the linker. 27052379 5 0 theme lower 1120:1124 arg1 affinity 1126:1133 a lower affinity 1118:1133 a lower affinity for its mono-α-D-glucoside product 1118:1168 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 1 1 theme acceptor 172:179 arg1 specificity 191:201 a broad acceptor substrate specificity 164:201 a broad acceptor substrate specificity 164:201 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 2 theme non-carbohydrate 282:297 arg1 molecules 299:307 small non-carbohydrate molecules 276:307 small non-carbohydrate molecules using sucrose as donor substrate 276:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 3 gly glycosylation 259:271 arg1 molecules 299:307 small non-carbohydrate molecules 276:307 small non-carbohydrate molecules using sucrose as donor substrate 276:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 4 theme substrate 181:189 arg1 specificity 191:201 a broad acceptor substrate specificity 164:201 a broad acceptor substrate specificity 164:201 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 5 theme molecules 299:307 arg1 glycosylation 259:271 the glycosylation 255:271 the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate 255:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 6 6 theme high 1247:1250 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 2 7 from synthesis 405:413 arg1 sucrose 448:454 sucrose 448:454 sucrose 448:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 5 8 theme mono-α-D-glucoside 1143:1160 arg1 product 1162:1168 its mono-α-D-glucoside product 1139:1168 its mono-α-D-glucoside product 1139:1168 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 6 9 theme clear 1320:1324 arg1 understanding 1326:1338 a clear understanding 1318:1338 a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential 1318:1483 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 3 10 theme α-glucan 641:648 arg1 synthesis 650:658 α-glucan synthesis 641:658 α-glucan synthesis 641:658 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 5 11 theme acceptor 1086:1093 arg1 catechol 1105:1112 the model acceptor substrate catechol 1076:1112 the model acceptor substrate catechol 1076:1112 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 0 12 theme improved 90:97 arg1 glycosylation 99:111 improved glycosylation 90:111 improved glycosylation of non-carbohydrate molecules 90:141 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 6 13 theme residues 1362:1369 arg1 mutagenesis 1347:1357 mutagenesis 1347:1357 mutagenesis of residues L938, L981, and N1029 1347:1391 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 4 14 dep mutants 823:829 arg1 mutants 823:829 Gtf180-ΔN mutants 813:829 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis 813:892 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 14 dep mutants 823:829 arg1 L981A 839:843 L981A 839:843 L981A 839:843 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 14 dep mutants 823:829 arg1 N1029M 850:855 N1029M 850:855 N1029M 850:855 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 14 dep mutants 823:829 arg1 L938F 832:836 L938F 832:836 L938F 832:836 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 2 15 theme glucansucrase-catalyzed 361:383 arg1 reaction 385:392 the main glucansucrase-catalyzed reaction 352:392 the main glucansucrase-catalyzed reaction 352:392 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 0 16 theme non-carbohydrate 116:131 arg1 molecules 133:141 non-carbohydrate molecules 116:141 non-carbohydrate molecules 116:141 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 4 17 with mutants 823:829 arg1 synthesis 884:892 an impaired α-glucan synthesis 863:892 an impaired α-glucan synthesis 863:892 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 6 18 with mutants 1436:1442 arg1 potential 1475:1483 an improved glycosylation potential 1449:1483 an improved glycosylation potential 1449:1483 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 6 19 dep residues 1362:1369 arg1 residues 1362:1369 residues L938, L981, and N1029 1362:1391 residues L938, L981, and N1029 1362:1391 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 6 19 dep residues 1362:1369 arg1 L981 1377:1380 L981 1377:1380 L981 1377:1380 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 6 19 dep residues 1362:1369 arg1 N1029 1387:1391 N1029 1387:1391 N1029 1387:1391 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 6 19 dep residues 1362:1369 arg1 L938 1371:1374 L938 1371:1374 L938 1371:1374 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 2 20 theme downstream 557:566 arg1 processing 568:577 downstream processing 557:577 downstream processing of glucosylated products 557:602 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 6 21 theme protein 1301:1307 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 4 22 theme α-glucan 875:882 arg1 synthesis 884:892 an impaired α-glucan synthesis 863:892 an impaired α-glucan synthesis 863:892 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 6 23 theme available 1237:1245 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 3 24 from construction 754:765 arg1 engineering 674:684 mutational engineering 663:684 mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts 663:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 2 25 theme non-carbohydrate 514:529 arg1 molecules 531:539 non-carbohydrate molecules 514:539 non-carbohydrate molecules 514:539 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 4 26 theme alcoholic 980:988 arg1 compounds 990:998 several phenolic and alcoholic compounds 959:998 compounds 990:998 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 0 27 theme reaction 65:72 arg1 engineering 74:84 reaction engineering 65:84 reaction engineering 65:84 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 2 28 theme main 356:359 arg1 reaction 385:392 the main glucansucrase-catalyzed reaction 352:392 the main glucansucrase-catalyzed reaction 352:392 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 5 29 theme Kinetic 1001:1007 arg1 analysis 1009:1016 Kinetic analysis 1001:1016 Kinetic analysis 1001:1016 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 0 30 theme Glucansucrase 0:12 arg1 Gtf180-ΔN 14:22 Glucansucrase Gtf180-ΔN 0:22 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 0:142 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 0 31 gly glycosylation 99:111 arg1 molecules 133:141 non-carbohydrate molecules 116:141 non-carbohydrate molecules 116:141 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 4 32 theme impaired 866:873 arg1 synthesis 884:892 an impaired α-glucan synthesis 863:892 an impaired α-glucan synthesis 863:892 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 1 33 theme increased 215:223 arg1 attention 225:233 increased attention 215:233 increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate 215:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 34 theme donor 326:330 arg1 substrate 332:340 donor substrate 326:340 donor substrate 326:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 6 35 theme Gtf180-ΔN 1291:1299 arg1 protein 1301:1307 the Gtf180-ΔN protein 1287:1307 the Gtf180-ΔN protein 1287:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 5 36 theme model 1080:1084 arg1 catechol 1105:1112 the model acceptor substrate catechol 1076:1112 the model acceptor substrate catechol 1076:1112 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 4 37 theme monoglycosylation 930:946 arg1 yields 948:953 monoglycosylation yields 930:953 monoglycosylation yields for several phenolic and alcoholic compounds 930:998 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 3 38 dep Lactobacillus 713:725 arg1 reuteri 727:733 reuteri 727:733 reuteri 727:733 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 2 39 theme efficient 487:495 arg1 glycosylation 497:509 the efficient glycosylation 483:509 the efficient glycosylation of non-carbohydrate molecules 483:539 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 0 40 theme molecules 133:141 arg1 glycosylation 99:111 improved glycosylation 90:111 improved glycosylation of non-carbohydrate molecules 90:141 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 3 41 theme mutational 663:672 arg1 engineering 674:684 mutational engineering 663:684 mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts 663:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 3 42 from enzyme 703:708 arg1 construction 754:765 the construction 750:765 the construction of more efficient glycosylation biocatalysts 750:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 4 43 theme Gtf180-ΔN 813:821 arg1 mutants 823:829 Gtf180-ΔN mutants 813:829 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis 813:892 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 43 theme Gtf180-ΔN 813:821 arg1 L981A 839:843 L981A 839:843 L981A 839:843 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 43 theme Gtf180-ΔN 813:821 arg1 N1029M 850:855 N1029M 850:855 N1029M 850:855 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 4 43 theme Gtf180-ΔN 813:821 arg1 L938F 832:836 L938F 832:836 L938F 832:836 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 0 44 theme 180 49:51 arg1 Gtf180-ΔN 14:22 Glucansucrase Gtf180-ΔN 0:22 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 0:142 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 5 45 theme substrate 1095:1103 arg1 catechol 1105:1112 the model acceptor substrate catechol 1076:1112 the model acceptor substrate catechol 1076:1112 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 3 46 theme enzyme 703:708 arg1 engineering 674:684 mutational engineering 663:684 mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts 663:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 6 47 theme 3D 1263:1264 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 6 48 theme α-glucan 1402:1409 arg1 synthesis 1411:1419 α-glucan synthesis 1402:1419 α-glucan synthesis 1402:1419 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 2 49 from sucrose 448:454 arg1 synthesis 405:413 synthesis 405:413 synthesis of α-glucan polysaccharides from sucrose 405:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 2 49 from sucrose 448:454 arg1 polysaccharides 427:441 α-glucan polysaccharides 418:441 α-glucan polysaccharides from sucrose 418:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 6 50 theme glycosylation 1461:1473 arg1 potential 1475:1483 an improved glycosylation potential 1449:1483 an improved glycosylation potential 1449:1483 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 5 51 theme improved 1186:1193 arg1 yields 1213:1218 the improved monoglycosylation yields 1182:1218 the improved monoglycosylation yields 1182:1218 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 6 52 theme resolution 1252:1261 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 2 53 theme molecules 531:539 arg1 glycosylation 497:509 the efficient glycosylation 483:509 the efficient glycosylation of non-carbohydrate molecules 483:539 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 6 54 theme improved 1452:1459 arg1 potential 1475:1483 an improved glycosylation potential 1449:1483 an improved glycosylation potential 1449:1483 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 3 55 theme glycosylation 785:797 arg1 biocatalysts 799:810 more efficient glycosylation biocatalysts 770:810 more efficient glycosylation biocatalysts 770:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 3 56 theme Gtf180-ΔN 693:701 arg1 enzyme 703:708 the Gtf180-ΔN enzyme 689:708 the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts 689:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 4 57 theme substantial 906:916 arg1 increase 918:925 a substantial increase 904:925 a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds 904:998 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 3 58 theme biocatalysts 799:810 arg1 construction 754:765 the construction 750:765 the construction of more efficient glycosylation biocatalysts 750:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 0 59 dep Lactobacillus 27:39 arg1 reuteri 41:47 reuteri 41:47 reuteri 41:47 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 5 60 contain possess 1046:1052 arg2 affinity 1063:1070 a higher affinity 1054:1070 a higher affinity for the model acceptor substrate catechol 1054:1112 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 5 60 contain possess 1046:1052 arg1 mutants 1038:1044 these mutants 1032:1044 these mutants 1032:1044 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 5 60 contain possess 1046:1052 arg2 affinity 1126:1133 a lower affinity 1118:1133 a lower affinity for its mono-α-D-glucoside product 1118:1168 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 5 61 theme higher 1056:1061 arg1 affinity 1063:1070 a higher affinity 1054:1070 a higher affinity for the model acceptor substrate catechol 1054:1112 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 3 62 theme Lactobacillus 713:725 arg1 results 739:745 180 results 735:745 Lactobacillus reuteri 180 results 713:745 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 6 63 theme crystal 1266:1272 arg1 structure 1274:1282 the available high resolution 3D crystal structure 1233:1282 the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1233:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 5 64 theme monoglycosylation 1195:1211 arg1 yields 1213:1218 the improved monoglycosylation yields 1182:1218 the improved monoglycosylation yields 1182:1218 Kinetic analysis revealed that these mutants possess a higher affinity for the model acceptor substrate catechol but a lower affinity for its mono-α-D-glucoside product, explaining the improved monoglycosylation yields. 27052379 3 65 theme results 739:745 arg1 enzyme 703:708 the Gtf180-ΔN enzyme 689:708 the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts 689:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 4 66 from increase 918:925 arg1 yields 948:953 monoglycosylation yields 930:953 monoglycosylation yields for several phenolic and alcoholic compounds 930:998 Gtf180-ΔN mutants (L938F, L981A, and N1029M) with an impaired α-glucan synthesis displayed a substantial increase in monoglycosylation yields for several phenolic and alcoholic compounds. 27052379 2 67 theme α-glucan 418:425 arg1 polysaccharides 427:441 α-glucan polysaccharides 418:441 α-glucan polysaccharides from sucrose 418:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 3 68 from engineering 674:684 arg1 construction 754:765 the construction 750:765 the construction of more efficient glycosylation biocatalysts 750:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 6 69 theme structure 1274:1282 arg1 Analysis 1221:1228 Analysis 1221:1228 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein 1221:1307 Analysis of the available high resolution 3D crystal structure of the Gtf180-ΔN protein provided a clear understanding of how mutagenesis of residues L938, L981, and N1029 impaired α-glucan synthesis, thus yielding mutants with an improved glycosylation potential. 27052379 2 70 theme products 595:602 arg1 processing 568:577 downstream processing 557:577 downstream processing of glucosylated products 557:602 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 3 71 theme efficient 775:783 arg1 biocatalysts 799:810 more efficient glycosylation biocatalysts 770:810 more efficient glycosylation biocatalysts 770:810 This paper reports that suppressing α-glucan synthesis by mutational engineering of the Gtf180-ΔN enzyme of Lactobacillus reuteri 180 results in the construction of more efficient glycosylation biocatalysts. 27052379 0 72 dep Gtf180-ΔN 14:22 arg1 engineering 74:84 reaction engineering 65:84 reaction engineering 65:84 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 0 72 dep Gtf180-ΔN 14:22 arg1 enzyme 54:59 enzyme 54:59 enzyme 54:59 Glucansucrase Gtf180-ΔN of Lactobacillus reuteri 180: enzyme and reaction engineering for improved glycosylation of non-carbohydrate molecules. 27052379 2 73 theme polysaccharides 427:441 arg1 synthesis 405:413 synthesis 405:413 synthesis of α-glucan polysaccharides from sucrose 405:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 2 74 theme glucosylated 582:593 arg1 products 595:602 glucosylated products 582:602 glucosylated products 582:602 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 1 75 contain have 159:162 arg1 Glucansucrases 144:157 Glucansucrases 144:157 Glucansucrases 144:157 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 75 contain have 159:162 arg2 specificity 191:201 a broad acceptor substrate specificity 164:201 a broad acceptor substrate specificity 164:201 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 2 76 gly glycosylation 497:509 arg1 molecules 531:539 non-carbohydrate molecules 514:539 non-carbohydrate molecules 514:539 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 2 76 gly glycosylation 497:509 arg1 synthesis 405:413 synthesis 405:413 synthesis of α-glucan polysaccharides from sucrose 405:454 However, the main glucansucrase-catalyzed reaction results in synthesis of α-glucan polysaccharides from sucrose, and this strongly impedes the efficient glycosylation of non-carbohydrate molecules and complicates downstream processing of glucosylated products. 27052379 1 77 theme broad 166:170 arg1 specificity 191:201 a broad acceptor substrate specificity 164:201 a broad acceptor substrate specificity 164:201 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 27052379 1 78 theme small 276:280 arg1 molecules 299:307 small non-carbohydrate molecules 276:307 small non-carbohydrate molecules using sucrose as donor substrate 276:340 Glucansucrases have a broad acceptor substrate specificity and receive increased attention as biocatalysts for the glycosylation of small non-carbohydrate molecules using sucrose as donor substrate. 25623399 2 0 theme receptor-activated 529:546 arg1 cells 550:554 T cell receptor-activated T cells 522:554 T cell receptor-activated T cells 522:554 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 3 1 theme immune 872:877 arg1 escape 879:884 immune escape 872:884 immune escape of glioma cells 872:900 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 2 2 theme T 522:522 arg1 cells 550:554 T cell receptor-activated T cells 522:554 T cell receptor-activated T cells 522:554 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 7 3 theme protein 1689:1695 arg1 yields 1697:1702 protein yields 1689:1702 protein yields 1689:1702 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 3 4 theme recognition 649:659 arg1 system 661:666 an alternative "missing-self" recognition system 619:666 an alternative "missing-self" recognition system 619:666 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 5 theme cells 896:900 arg1 escape 879:884 immune escape 872:884 immune escape of glioma cells 872:900 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 4 6 theme expression 1024:1033 arg1 system 1035:1040 an eukaryotic expression system 1010:1040 an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1010:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 7 theme missing-self 635:646 arg1 system 661:666 an alternative "missing-self" recognition system 619:666 an alternative "missing-self" recognition system 619:666 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 4 8 dep line 1091:1094 arg1 - 1083:1083 - 1083:1083 - 1083:1083 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 1 9 theme Lectin-like 177:187 arg1 ligand 245:250 a ligand 243:250 a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes 243:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 1 9 theme Lectin-like 177:187 arg1 transcript 189:198 Lectin-like transcript 1 177:200 Lectin-like transcript 1 (LLT1, gene clec2d) 177:220 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 2 10 theme dendritic 491:499 arg1 cells 501:505 TLR-stimulated dendritic cells 476:505 TLR-stimulated dendritic cells 476:505 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 7 11 theme density 1640:1646 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 4 12 theme ectodomain 996:1005 arg1 mutant 981:986 soluble His176Cys mutant 963:986 soluble His176Cys mutant of LLT1 ectodomain 963:1005 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 7 13 theme GnTI 1523:1526 arg1 line 1536:1539 HEK293S GnTI(-) cell line 1515:1539 HEK293S GnTI(-) cell line 1515:1539 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 13 theme GnTI 1523:1526 arg1 part 1463:1466 a part 1461:1466 a part of production of soluble LLT1 1461:1496 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 5 14 from reconstruction 1211:1224 arg1 LLT1 1265:1268 LLT1 1265:1268 LLT1 1265:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 0 15 theme suspension 135:144 arg1 transfection 119:130 high-density transfection 106:130 high-density transfection of suspension 106:144 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 0 16 theme HEK293S 154:160 arg1 GnTI 162:165 HEK293S GnTI 154:165 HEK293S GnTI 154:165 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 7 17 theme novel 1624:1628 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 8 18 theme protein 1817:1823 arg1 crystallography 1825:1839 protein crystallography 1817:1839 protein crystallography 1817:1839 This transfection protocol is generally applicable for protein production within this cell line, especially for protein crystallography. 25623399 7 19 from growth 1544:1549 arg1 media 1568:1572 media 1568:1572 media facilitating transient transfection and optimized novel high cell density transfection protocol 1568:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 19 from growth 1544:1549 arg1 suspension 1554:1563 suspension 1554:1563 suspension 1554:1563 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 4 20 theme cell 1086:1089 arg1 line 1091:1094 human suspension-adapted HEK293S GnTI(-) cell line 1045:1094 human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1045:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 21 theme NK 734:735 arg1 cells 760:764 NK and antigen presenting cells 734:764 cells 760:764 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 7 22 theme LLT1 1493:1496 arg1 production 1471:1480 production 1471:1480 production of soluble LLT1 1471:1496 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 4 23 theme suspension-adapted 1051:1068 arg1 line 1091:1094 human suspension-adapted HEK293S GnTI(-) cell line 1045:1094 human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1045:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 24 theme crosstalk 716:724 arg1 regulation 702:711 regulation 702:711 regulation of crosstalk between NK and antigen presenting cells 702:764 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 1 25 theme present 289:295 arg1 receptor 280:287 the single human NKR-P1 receptor 256:287 the single human NKR-P1 receptor present on NK and NK-T lymphocytes 256:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 4 26 theme transient 918:926 arg1 expression 928:937 expression 928:937 expression 928:937 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 7 27 theme transient 1587:1595 arg1 transfection 1597:1608 transient transfection 1587:1608 transient transfection 1587:1608 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 4 28 theme GnTI 1078:1081 arg1 line 1091:1094 human suspension-adapted HEK293S GnTI(-) cell line 1045:1094 human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1045:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 0 29 theme killer 72:77 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 3 30 theme glioblastoma 790:801 arg1 cells 803:807 glioblastoma cells 790:807 glioblastoma cells 790:807 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 30 theme glioblastoma 790:801 arg1 tumors 833:838 the most lethal tumors 817:838 the most lethal tumors 817:838 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 30 theme glioblastoma 790:801 arg1 one 810:812 one 810:812 one 810:812 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 2 31 theme cells 464:468 arg1 activation 437:446 activation 437:446 activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells 437:554 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 5 32 theme lectin-like 1166:1176 arg1 stability 1185:1193 C-type lectin-like domain stability 1159:1193 C-type lectin-like domain stability 1159:1193 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 3 33 theme antigen 741:747 arg1 cells 760:764 NK and antigen presenting cells 734:764 cells 760:764 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 0 34 theme receptor 84:91 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 8 35 theme transfection 1710:1721 arg1 protocol 1723:1730 This transfection protocol 1705:1730 This transfection protocol 1705:1730 This transfection protocol is generally applicable for protein production within this cell line, especially for protein crystallography. 25623399 2 36 theme other 451:455 arg1 cells 464:468 other immune cells 451:468 other immune cells 451:468 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 5 37 theme canonical 1235:1243 arg1 bridge 1255:1260 third canonical disulfide bridge 1229:1260 third canonical disulfide bridge in LLT1 1229:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 5 38 theme bridge 1255:1260 arg1 reconstruction 1211:1224 the reconstruction 1207:1224 the reconstruction of third canonical disulfide bridge in LLT1 1207:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 4 39 from expression 928:937 arg1 system 1035:1040 an eukaryotic expression system 1010:1040 an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1010:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 5 40 from LLT1 1265:1268 arg1 reconstruction 1211:1224 the reconstruction 1207:1224 the reconstruction of third canonical disulfide bridge in LLT1 1207:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 2 41 theme cells 375:379 arg1 surface 361:367 the surface 357:367 the surface of NK cells 357:379 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 3 42 theme tumors 833:838 arg1 cells 803:807 glioblastoma cells 790:807 glioblastoma cells 790:807 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 42 theme tumors 833:838 arg1 tumors 833:838 the most lethal tumors 817:838 the most lethal tumors 817:838 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 42 theme tumors 833:838 arg1 one 810:812 one 810:812 one 810:812 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 0 43 theme soluble 31:37 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 0 43 theme soluble 31:37 arg1 ectodomain 50:59 soluble and stable ectodomain 31:59 soluble and stable ectodomain of natural killer cell receptor LLT1 31:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 1 44 theme single 260:265 arg1 receptor 280:287 the single human NKR-P1 receptor 256:287 the single human NKR-P1 receptor present on NK and NK-T lymphocytes 256:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 6 45 theme soluble 1311:1317 arg1 LLT1 1319:1322 Purified soluble LLT1 1302:1322 Purified soluble LLT1 1302:1322 Purified soluble LLT1 is homogeneous, deglycosylatable and forms a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans. 25623399 3 46 theme normal 566:571 arg1 tissues 573:579 normal tissues 566:579 normal tissues 566:579 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 0 47 theme stable 43:48 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 0 47 theme stable 43:48 arg1 ectodomain 50:59 soluble and stable ectodomain 31:59 soluble and stable ectodomain of natural killer cell receptor LLT1 31:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 1 48 theme NKR-P1 273:278 arg1 receptor 280:287 the single human NKR-P1 receptor 256:287 the single human NKR-P1 receptor present on NK and NK-T lymphocytes 256:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 4 49 theme mutant 981:986 arg1 characterization 943:958 characterization 943:958 characterization 943:958 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 4 49 theme mutant 981:986 arg1 expression 928:937 expression 928:937 expression 928:937 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 50 theme escape 879:884 arg1 mediator 860:867 a mediator 858:867 a mediator of immune escape of glioma cells 858:900 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 50 theme escape 879:884 arg1 it 847:848 it 847:848 it 847:848 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 4 51 theme LLT1 991:994 arg1 ectodomain 996:1005 LLT1 ectodomain 991:1005 LLT1 ectodomain 991:1005 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 52 theme NKR-P1 586:591 arg1 interaction 593:603 NKR-P1 interaction 586:603 LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) 581:667 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 1 53 dep transcript 189:198 arg1 clec2d 214:219 gene clec2d 209:219 gene clec2d 209:219 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 1 53 dep transcript 189:198 arg1 LLT1 203:206 LLT1 203:206 LLT1 203:206 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 2 54 dep cells 501:505 arg1 e.g. 471:474 e.g. 471:474 e.g. 471:474 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 8 55 theme protein 1760:1766 arg1 production 1768:1777 protein production 1760:1777 protein production within this cell line 1760:1799 This transfection protocol is generally applicable for protein production within this cell line, especially for protein crystallography. 25623399 2 56 theme cell 524:527 arg1 cells 550:554 T cell receptor-activated T cells 522:554 T cell receptor-activated T cells 522:554 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 4 57 theme soluble 963:969 arg1 mutant 981:986 soluble His176Cys mutant 963:986 soluble His176Cys mutant of LLT1 ectodomain 963:1005 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 58 dep interaction 593:603 arg1 representing 606:617 representing 606:617 representing an alternative "missing-self" recognition system 606:666 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 3 59 theme glioma 889:894 arg1 cells 896:900 glioma cells 889:900 glioma cells 889:900 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 8 60 theme cell 1791:1794 arg1 line 1796:1799 this cell line 1786:1799 this cell line 1786:1799 This transfection protocol is generally applicable for protein production within this cell line, especially for protein crystallography. 25623399 6 61 theme N-glycans 1447:1455 arg1 presence 1431:1438 presence 1431:1438 presence of its N-glycans 1431:1455 Purified soluble LLT1 is homogeneous, deglycosylatable and forms a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans. 25623399 7 62 theme transfection 1648:1659 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 4 63 theme human 1045:1049 arg1 line 1091:1094 human suspension-adapted HEK293S GnTI(-) cell line 1045:1094 human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1045:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 64 theme " 647:647 arg1 system 661:666 an alternative "missing-self" recognition system 619:666 an alternative "missing-self" recognition system 619:666 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 2 65 theme T 548:548 arg1 cells 550:554 T cell receptor-activated T cells 522:554 T cell receptor-activated T cells 522:554 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 7 66 theme cell 1635:1638 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 3 67 theme immunomodulatory 677:692 arg1 role 694:697 an immunomodulatory role 674:697 an immunomodulatory role 674:697 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 7 68 theme HEK293S 1515:1521 arg1 GnTI 1523:1526 HEK293S GnTI 1515:1526 HEK293S GnTI 1515:1526 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 2 69 theme TLR-stimulated 476:489 arg1 cells 501:505 TLR-stimulated dendritic cells 476:505 TLR-stimulated dendritic cells 476:505 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 4 70 theme eukaryotic 1013:1022 arg1 system 1035:1040 an eukaryotic expression system 1010:1040 an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1010:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 0 71 theme GnTI 162:165 arg1 cells 170:174 HEK293S GnTI(-) cells 154:174 HEK293S GnTI(-) cells 154:174 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 6 72 theme Purified 1302:1309 arg1 LLT1 1319:1322 Purified soluble LLT1 1302:1322 Purified soluble LLT1 1302:1322 Purified soluble LLT1 is homogeneous, deglycosylatable and forms a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans. 25623399 2 73 theme B 511:511 arg1 cells 513:517 B cells 511:517 B cells 511:517 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 4 74 theme line 1091:1094 arg1 system 1035:1040 an eukaryotic expression system 1010:1040 an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1010:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 5 75 theme domain 1178:1183 arg1 stability 1185:1193 C-type lectin-like domain stability 1159:1193 C-type lectin-like domain stability 1159:1193 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 7 76 theme high 1630:1633 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 77 theme soluble 1485:1491 arg1 LLT1 1493:1496 soluble LLT1 1485:1496 soluble LLT1 1485:1496 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 5 78 from bridge 1255:1260 arg1 LLT1 1265:1268 LLT1 1265:1268 LLT1 1265:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 4 79 theme HEK293S 1070:1076 arg1 line 1091:1094 human suspension-adapted HEK293S GnTI(-) cell line 1045:1094 human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1045:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 0 80 theme natural 64:70 arg1 killer 72:77 natural killer 64:77 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 7 81 theme optimized 1614:1622 arg1 protocol 1661:1668 optimized novel high cell density transfection protocol 1614:1668 optimized novel high cell density transfection protocol 1614:1668 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 82 theme production 1471:1480 arg1 part 1463:1466 a part 1461:1466 a part of production of soluble LLT1 1461:1496 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 82 theme production 1471:1480 arg1 line 1536:1539 HEK293S GnTI(-) cell line 1515:1539 HEK293S GnTI(-) cell line 1515:1539 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 0 83 theme cell 79:82 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 4 84 from characterization 943:958 arg1 system 1035:1040 an eukaryotic expression system 1010:1040 an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans 1010:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 5 85 theme C-type 1159:1164 arg1 stability 1185:1193 C-type lectin-like domain stability 1159:1193 C-type lectin-like domain stability 1159:1193 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 0 86 theme LLT1 93:96 arg1 LLT1 93:96 natural killer cell receptor LLT1 64:96 natural killer cell receptor LLT1 64:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 0 86 theme LLT1 93:96 arg1 ectodomain 50:59 soluble and stable ectodomain 31:59 soluble and stable ectodomain of natural killer cell receptor LLT1 31:96 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 2 87 theme immune 457:462 arg1 cells 464:468 other immune cells 451:468 other immune cells 451:468 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 5 88 theme third 1229:1233 arg1 bridge 1255:1260 third canonical disulfide bridge 1229:1260 third canonical disulfide bridge in LLT1 1229:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 3 89 theme presenting 749:758 arg1 cells 760:764 NK and antigen presenting cells 734:764 cells 760:764 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 0 90 theme high-density 106:117 arg1 transfection 119:130 high-density transfection 106:130 high-density transfection of suspension 106:144 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 3 91 theme lethal 826:831 arg1 tumors 833:838 the most lethal tumors 817:838 the most lethal tumors 817:838 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 4 92 theme uniform 1101:1107 arg1 N-glycans 1109:1117 uniform N-glycans 1101:1117 uniform N-glycans 1101:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 1 93 theme gene 209:212 arg1 clec2d 214:219 gene clec2d 209:219 gene clec2d 209:219 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 1 93 theme gene 209:212 arg1 LLT1 203:206 LLT1 203:206 LLT1 203:206 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 5 94 theme disulfide 1245:1253 arg1 bridge 1255:1260 third canonical disulfide bridge 1229:1260 third canonical disulfide bridge in LLT1 1229:1268 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 1 95 dep NK 300:301 arg1 lymphocytes 312:322 lymphocytes 312:322 lymphocytes 312:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 2 96 theme NK 372:373 arg1 cells 375:379 NK cells 372:379 NK cells 372:379 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 7 97 theme cell 1531:1534 arg1 line 1536:1539 HEK293S GnTI(-) cell line 1515:1539 HEK293S GnTI(-) cell line 1515:1539 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 7 97 theme cell 1531:1534 arg1 part 1463:1466 a part 1461:1466 a part of production of soluble LLT1 1461:1496 As a part of production of soluble LLT1, we have adapted HEK293S GnTI(-) cell line to growth in suspension in media facilitating transient transfection and optimized novel high cell density transfection protocol, greatly enhancing protein yields. 25623399 0 98 theme ectodomain 50:59 arg1 purification 15:26 purification 15:26 purification 15:26 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 0 98 theme ectodomain 50:59 arg1 Expression 0:9 Expression 0:9 Expression 0:9 Expression and purification of soluble and stable ectodomain of natural killer cell receptor LLT1 through high-density transfection of suspension adapted HEK293S GnTI(-) cells. 25623399 1 99 theme human 267:271 arg1 receptor 280:287 the single human NKR-P1 receptor 256:287 the single human NKR-P1 receptor present on NK and NK-T lymphocytes 256:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 6 100 dep homogeneous 1327:1337 arg1 deglycosylatable 1340:1355 deglycosylatable 1340:1355 deglycosylatable 1340:1355 Purified soluble LLT1 is homogeneous, deglycosylatable and forms a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans. 25623399 2 101 theme IFN-γ 394:398 arg1 production 400:409 IFN-γ production 394:409 IFN-γ production 394:409 Naturally, LLT1 is expressed on the surface of NK cells, stimulating IFN-γ production, and is up-regulated upon activation of other immune cells, e.g. TLR-stimulated dendritic cells and B cells or T cell receptor-activated T cells. 25623399 5 102 theme mass 1283:1286 arg1 spectrometry 1288:1299 mass spectrometry 1283:1299 mass spectrometry 1283:1299 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 1 103 attach present 289:295 arg2 receptor 280:287 the single human NKR-P1 receptor 256:287 the single human NKR-P1 receptor present on NK and NK-T lymphocytes 256:322 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 1 103 attach present 289:295 arg1 NK 300:301 NK 300:301 NK 300:301 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 1 103 attach present 289:295 arg1 NK-T 307:310 NK-T 307:310 NK-T 307:310 Lectin-like transcript 1 (LLT1, gene clec2d) was identified to be a ligand for the single human NKR-P1 receptor present on NK and NK-T lymphocytes. 25623399 4 104 theme His176Cys 971:979 arg1 mutant 981:986 soluble His176Cys mutant 963:986 soluble His176Cys mutant of LLT1 ectodomain 963:1005 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 5 105 theme His176Cys 1124:1132 arg1 critical 1146:1153 critical 1146:1153 critical 1146:1153 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 5 105 theme His176Cys 1124:1132 arg1 mutation 1134:1141 The His176Cys mutation 1120:1141 The His176Cys mutation 1120:1141 The His176Cys mutation is critical for C-type lectin-like domain stability, leading to the reconstruction of third canonical disulfide bridge in LLT1, as shown by mass spectrometry. 25623399 4 106 with line 1091:1094 arg1 N-glycans 1109:1117 uniform N-glycans 1101:1117 uniform N-glycans 1101:1117 Here we report transient expression and characterization of soluble His176Cys mutant of LLT1 ectodomain in an eukaryotic expression system of human suspension-adapted HEK293S GnTI(-) cell line with uniform N-glycans. 25623399 3 107 theme alternative 622:632 arg1 system 661:666 an alternative "missing-self" recognition system 619:666 an alternative "missing-self" recognition system 619:666 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25623399 6 108 theme non-covalent 1369:1380 arg1 homodimer 1382:1390 a non-covalent homodimer 1367:1390 a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans 1367:1455 Purified soluble LLT1 is homogeneous, deglycosylatable and forms a non-covalent homodimer whose dimerization is not dependent on presence of its N-glycans. 25623399 3 109 dep LLT1 581:584 arg1 interaction 593:603 NKR-P1 interaction 586:603 LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) 581:667 While in normal tissues LLT1:NKR-P1 interaction (representing an alternative "missing-self" recognition system) play an immunomodulatory role in regulation of crosstalk between NK and antigen presenting cells, LLT1 is upregulated in glioblastoma cells, one of the most lethal tumors, where it acts as a mediator of immune escape of glioma cells. 25880336 10 0 theme form 1292:1295 arg1 prevalence 1273:1282 prevalence 1273:1282 prevalence of this form of the sugar in the OS-derived species 1273:1334 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 3 1 dep NMR 349:351 arg1 methods 375:381 methods 375:381 methods 375:381 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 11 2 theme R. 1504:1505 arg1 leguminosarum 1507:1519 R. leguminosarum bv 1504:1522 R. leguminosarum bv 1504:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 12 3 theme α-L-Rhap-2-OAc 1562:1575 arg1 * 1576:1576 α-L-Rhap-2-OAc* 1562:1576 α-L-Rhap-2-OAc* 1562:1576 trifolii strain 24, was established: α-L-Rhap-2-OAc*-(1-->3)-α-L-Rhap-(1-->3)-β-D-QuipNAc-(1-->4)-Kdo * ~ 50%. 25880336 11 4 theme following 1369:1377 arg1 structure 1379:1387 the following structure 1365:1387 the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv 1365:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 11 5 theme leguminosarum 1507:1519 arg1 LPS 1497:1499 the LPS 1493:1499 the LPS of R. leguminosarum bv 1493:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 7 6 theme core 843:846 arg1 OS 848:849 the core OS 839:849 the core OS 839:849 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 5 7 theme molecule 619:626 arg1 mass 628:631 the higher molecule mass 608:631 the higher molecule mass 608:631 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 0 8 theme leguminosarum 92:104 arg1 LPS 75:77 the LPS 71:77 the LPS of Rhizobium leguminosarum bv 71:107 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 9 9 located detected 1036:1043 arg2 result 1121:1126 a result 1119:1126 a result of Smith degradation 1119:1147 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 9 located detected 1036:1043 arg2 acid 1014:1017 3-deoxyheptulosonic acid 994:1017 3-deoxyheptulosonic acid (Kdh) 994:1023 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 9 located detected 1036:1043 arg2 Kdh 1020:1022 Kdh 1020:1022 Kdh 1020:1022 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 9 located detected 1036:1043 arg1 terminus 1061:1068 the reducing terminus 1048:1068 the reducing terminus of the OS 1048:1078 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 4 10 from 728.3 476:480 arg1 ions 454:457 two molecular ions 440:457 two molecular ions 440:457 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 8 11 theme terminal 939:946 arg1 Rhap 948:951 the terminal Rhap 935:951 the terminal Rhap 935:951 2D NMR experiments revealed that the terminal Rhap is acetylated at position O-2. 25880336 7 12 theme OS 848:849 arg1 composition 798:808 composition 798:808 composition 798:808 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 7 12 theme OS 848:849 arg1 pattern 828:834 glycosylation pattern 814:834 glycosylation pattern 814:834 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 0 13 theme Rhizobium 82:90 arg1 leguminosarum 92:104 Rhizobium leguminosarum bv 82:107 Rhizobium leguminosarum bv 82:107 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 4 14 theme electrospray 397:408 arg1 spectrum 424:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 10 15 theme pyranose 1214:1221 arg1 ring 1223:1226 a six-membered pyranose ring 1199:1226 a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf 1199:1252 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 3 16 theme methylation 317:327 arg1 analyses 329:336 sugar and methylation analyses 307:336 analyses 329:336 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 2 17 theme core 140:143 arg1 OS 162:163 OS 162:163 OS 162:163 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 17 theme core 140:143 arg1 oligosaccharide 145:159 The outer core oligosaccharide 130:159 The outer core oligosaccharide (OS) 130:164 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 4 18 theme Negative-ion 384:395 arg1 spectrum 424:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 9 19 theme 3-deoxyheptulosonic 994:1012 arg1 result 1121:1126 a result 1119:1126 a result of Smith degradation 1119:1147 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 19 theme 3-deoxyheptulosonic 994:1012 arg1 Kdh 1020:1022 Kdh 1020:1022 Kdh 1020:1022 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 19 theme 3-deoxyheptulosonic 994:1012 arg1 acid 1014:1017 3-deoxyheptulosonic acid 994:1017 3-deoxyheptulosonic acid (Kdh) 994:1023 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 2 20 theme outer 134:138 arg1 OS 162:163 OS 162:163 OS 162:163 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 20 theme outer 134:138 arg1 oligosaccharide 145:159 The outer core oligosaccharide 130:159 The outer core oligosaccharide (OS) 130:164 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 4 21 from m/z 462:464 arg1 ions 454:457 two molecular ions 440:457 two molecular ions 440:457 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 8 22 theme position 970:977 arg1 O-2 979:981 position O-2 970:981 position O-2 970:981 2D NMR experiments revealed that the terminal Rhap is acetylated at position O-2. 25880336 7 23 theme glycosylation 814:826 arg1 pattern 828:834 glycosylation pattern 814:834 glycosylation pattern 814:834 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 11 24 attach links 1439:1443 arg2 tetrasaccharide 1407:1421 the outer core tetrasaccharide 1392:1421 the outer core tetrasaccharide 1392:1421 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 11 24 attach links 1439:1443 arg1 core 1485:1488 the inner core 1475:1488 the inner core in the LPS of R. leguminosarum bv 1475:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 3 25 theme mass 357:360 arg1 spectrometry 362:373 mass spectrometry 357:373 mass spectrometry 357:373 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 7 26 gly glycosylation 814:826 arg1 OS 848:849 the core OS 839:849 the core OS 839:849 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 9 27 theme Smith 1131:1135 arg1 degradation 1137:1147 Smith degradation 1131:1147 Smith degradation 1131:1147 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 2 28 attach isolated 170:177 arg2 oligosaccharide 145:159 The outer core oligosaccharide 130:159 The outer core oligosaccharide (OS) 130:164 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 28 attach isolated 170:177 arg1 LPS 208:210 LPS 208:210 LPS 208:210 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 28 attach isolated 170:177 arg2 OS 162:163 OS 162:163 OS 162:163 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 28 attach isolated 170:177 arg1 lipopolysaccharide 188:205 the lipopolysaccharide 184:205 the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv 184:241 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 0 29 theme Structural 0:9 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of the outer core tetrasaccharide 0:55 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 7 30 theme NMR 767:769 arg1 spectroscopy 771:782 NMR spectroscopy 767:782 NMR spectroscopy 767:782 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 5 31 theme higher 612:617 arg1 mass 628:631 the higher molecule mass 608:631 the higher molecule mass 608:631 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 9 32 theme degradation 1137:1147 arg1 result 1121:1126 a result 1119:1126 a result of Smith degradation 1119:1147 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 32 theme degradation 1137:1147 arg1 acid 1014:1017 3-deoxyheptulosonic acid 994:1017 3-deoxyheptulosonic acid (Kdh) 994:1023 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 7 33 from evidence 875:882 arg1 structure 891:899 its structure 887:899 its structure 887:899 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 12 34 theme trifolii 1525:1532 arg1 strain 1534:1539 trifolii strain 24 1525:1542 trifolii strain 24 1525:1542 trifolii strain 24, was established: α-L-Rhap-2-OAc*-(1-->3)-α-L-Rhap-(1-->3)-β-D-QuipNAc-(1-->4)-Kdo * ~ 50%. 25880336 10 35 theme OS-derived 1317:1326 arg1 species 1328:1334 the OS-derived species 1313:1334 the OS-derived species 1313:1334 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 5 36 theme mass 559:562 arg1 difference 564:573 The mass difference 555:573 The mass difference between both ions 555:591 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 9 37 theme reducing 1052:1059 arg1 terminus 1061:1068 the reducing terminus 1048:1068 the reducing terminus of the OS 1048:1078 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 11 38 from core 1485:1488 arg1 LPS 1497:1499 the LPS 1493:1499 the LPS of R. leguminosarum bv 1493:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 11 39 theme inner 1479:1483 arg1 core 1485:1488 the inner core 1475:1488 the inner core in the LPS of R. leguminosarum bv 1475:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 4 40 theme composition 528:538 arg1 Rha2QuiNAcKdh 540:552 the composition Rha2QuiNAcKdh 524:552 the composition Rha2QuiNAcKdh 524:552 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 5 41 theme OS 682:683 arg1 variant 667:673 the mono O-acetylated variant 645:673 the mono O-acetylated variant of the OS 645:683 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 4 42 contain having 517:522 arg2 Rha2QuiNAcKdh 540:552 the composition Rha2QuiNAcKdh 524:552 the composition Rha2QuiNAcKdh 524:552 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 4 42 contain having 517:522 arg1 OS 514:515 the core OS 505:515 the core OS having the composition Rha2QuiNAcKdh 505:552 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 11 43 dep leguminosarum 1507:1519 arg1 bv 1521:1522 bv 1521:1522 bv 1521:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 10 44 from prevalence 1273:1282 arg1 species 1328:1334 the OS-derived species 1313:1334 the OS-derived species 1313:1334 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 0 45 theme core 36:39 arg1 tetrasaccharide 41:55 the outer core tetrasaccharide 26:55 the outer core tetrasaccharide 26:55 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 10 46 theme six-membered 1201:1212 arg1 ring 1223:1226 a six-membered pyranose ring 1199:1226 a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf 1199:1252 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 3 47 theme trifolii 244:251 arg1 strain 253:258 trifolii strain 24 244:261 trifolii strain 24 after Smith degradation 244:285 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 11 48 theme core 1402:1405 arg1 tetrasaccharide 1407:1421 the outer core tetrasaccharide 1392:1421 the outer core tetrasaccharide 1392:1421 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 11 49 theme O-chain 1449:1455 arg1 polysaccharide 1457:1470 the O-chain polysaccharide 1445:1470 the O-chain polysaccharide 1445:1470 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 0 50 theme outer 30:34 arg1 tetrasaccharide 41:55 the outer core tetrasaccharide 26:55 the outer core tetrasaccharide 26:55 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 9 51 attach derived 1095:1101 arg2 result 1121:1126 a result 1119:1126 a result of Smith degradation 1119:1147 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 51 attach derived 1095:1101 arg1 Kdo 1112:1114 the Kdo 1108:1114 the Kdo 1108:1114 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 51 attach derived 1095:1101 arg2 acid 1014:1017 3-deoxyheptulosonic acid 994:1017 3-deoxyheptulosonic acid (Kdh) 994:1023 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 9 51 attach derived 1095:1101 arg2 Kdh 1020:1022 Kdh 1020:1022 Kdh 1020:1022 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 12 52 theme -Kdo 1622:1625 arg1 * 1627:1627 (1-->3)-α-L-Rhap-(1-->3)-β-D-QuipNAc-(1-->4)-Kdo * 1578:1627 (1-->3)-α-L-Rhap-(1-->3)-β-D-QuipNAc-(1-->4)-Kdo * 1578:1627 trifolii strain 24, was established: α-L-Rhap-2-OAc*-(1-->3)-α-L-Rhap-(1-->3)-β-D-QuipNAc-(1-->4)-Kdo * ~ 50%. 25880336 4 53 theme molecular 444:452 arg1 ions 454:457 two molecular ions 440:457 two molecular ions 440:457 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 6 54 theme CID 739:741 arg1 spectra 749:755 CID MS/MS spectra 739:755 CID MS/MS spectra 739:755 The sequence of the oligosaccharide was reflected in CID MS/MS spectra. 25880336 10 55 link OS-derived 1317:1326 arg1 species 1328:1334 the OS-derived species 1313:1334 the OS-derived species 1313:1334 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 5 56 theme mono 649:652 arg1 variant 667:673 the mono O-acetylated variant 645:673 the mono O-acetylated variant of the OS 645:683 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 10 57 theme sugar 1304:1308 arg1 form 1292:1295 this form 1287:1295 this form of the sugar 1287:1308 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 9 58 theme OS 1077:1078 arg1 terminus 1061:1068 the reducing terminus 1048:1068 the reducing terminus of the OS 1048:1078 Moreover, 3-deoxyheptulosonic acid (Kdh), which was detected at the reducing terminus of the OS, was evidently derived from the Kdo as a result of Smith degradation. 25880336 1 59 theme trifolii 110:117 arg1 strain 119:124 trifolii strain 24 110:127 trifolii strain 24. 110:128 trifolii strain 24. 25880336 0 60 theme tetrasaccharide 41:55 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of the outer core tetrasaccharide 0:55 Structural elucidation of the outer core tetrasaccharide isolated from the LPS of Rhizobium leguminosarum bv. 25880336 4 61 theme core 509:512 arg1 OS 514:515 the core OS 505:515 the core OS having the composition Rha2QuiNAcKdh 505:552 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 5 62 theme O-acetylated 654:665 arg1 variant 667:673 the mono O-acetylated variant 645:673 the mono O-acetylated variant of the OS 645:683 The mass difference between both ions indicated that the higher molecule mass represented the mono O-acetylated variant of the OS. 25880336 4 63 theme ESI-MS 411:416 arg1 spectrum 424:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 8 64 theme NMR 905:907 arg1 experiments 909:919 2D NMR experiments 902:919 2D NMR experiments 902:919 2D NMR experiments revealed that the terminal Rhap is acetylated at position O-2. 25880336 6 65 theme MS/MS 743:747 arg1 spectra 749:755 CID MS/MS spectra 739:755 CID MS/MS spectra 739:755 The sequence of the oligosaccharide was reflected in CID MS/MS spectra. 25880336 3 66 theme Smith 269:273 arg1 degradation 275:285 Smith degradation 269:285 Smith degradation 269:285 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 8 67 theme 2D 902:903 arg1 experiments 909:919 2D NMR experiments 902:919 2D NMR experiments 902:919 2D NMR experiments revealed that the terminal Rhap is acetylated at position O-2. 25880336 11 68 theme outer 1396:1400 arg1 tetrasaccharide 1407:1421 the outer core tetrasaccharide 1392:1421 the outer core tetrasaccharide 1392:1421 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 4 69 theme mass 419:422 arg1 spectrum 424:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum 384:431 Negative-ion electrospray (ESI-MS) mass spectrum showed two molecular ions at m/z 686.3 and 728.3, which corresponded to the core OS having the composition Rha2QuiNAcKdh. 25880336 6 70 theme oligosaccharide 706:720 arg1 sequence 690:697 The sequence 686:697 The sequence of the oligosaccharide 686:720 The sequence of the oligosaccharide was reflected in CID MS/MS spectra. 25880336 10 71 theme Kdhp 1231:1234 arg1 ring 1223:1226 a six-membered pyranose ring 1199:1226 a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf 1199:1252 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 7 72 theme additional 864:873 arg1 evidence 875:882 additional evidence 864:882 additional evidence on its structure 864:899 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 3 73 theme sugar 307:311 arg1 analyses 329:336 sugar and methylation analyses 307:336 analyses 329:336 trifolii strain 24 after Smith degradation and then studied by sugar and methylation analyses along with NMR and mass spectrometry methods. 25880336 10 74 theme higher 1167:1172 arg1 intensity 1174:1182 the higher intensity 1163:1182 the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf 1163:1252 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 25880336 2 75 theme leguminosarum 226:238 arg1 LPS 208:210 LPS 208:210 LPS 208:210 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 2 75 theme leguminosarum 226:238 arg1 lipopolysaccharide 188:205 the lipopolysaccharide 184:205 the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv 184:241 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 11 76 theme tetrasaccharide 1407:1421 arg1 structure 1379:1387 the following structure 1365:1387 the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv 1365:1522 Based on the data obtained, the following structure of the outer core tetrasaccharide, which probably links the O-chain polysaccharide to the inner core in the LPS of R. leguminosarum bv. 25880336 2 77 theme Rhizobium 216:224 arg1 leguminosarum 226:238 Rhizobium leguminosarum bv 216:241 Rhizobium leguminosarum bv 216:241 The outer core oligosaccharide (OS) was isolated from the lipopolysaccharide (LPS) of Rhizobium leguminosarum bv. 25880336 7 78 dep composition 798:808 arg1 the 794:796 the 794:796 the 794:796 In turn, NMR spectroscopy confirmed the composition and glycosylation pattern of the core OS and provided additional evidence on its structure. 25880336 10 79 theme signals 1187:1193 arg1 intensity 1174:1182 the higher intensity 1163:1182 the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf 1163:1252 In addition, the higher intensity of signals for a six-membered pyranose ring of Kdhp over 2,7-anh-Kdhf seemed to indicate prevalence of this form of the sugar in the OS-derived species. 29076618 2 0 theme Gram-negative 119:131 arg1 bacteria 133:140 various Gram-negative bacteria 111:140 various Gram-negative bacteria 111:140 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 7 1 theme X-ray 1017:1021 arg1 analysis 1033:1040 Our X-ray structure analysis 1013:1040 Our X-ray structure analysis 1013:1040 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 4 2 located found 496:500 arg2 Mafs 490:493 Mafs 490:493 Mafs 490:493 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 2 located found 496:500 arg2 factors 481:487 associated factors 470:487 associated factors (Mafs) found in many pathogenic strains 470:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 2 located found 496:500 arg1 strains 521:527 many pathogenic strains 505:527 many pathogenic strains 505:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 6 3 theme nonflagellated 921:934 arg1 bacterium 936:944 a nonflagellated bacterium 919:944 a nonflagellated bacterium where the flagellin was still produced but no longer glycosylated 919:1010 Deletion of the gene amb0685 coding for Maf produced a nonflagellated bacterium where the flagellin was still produced but no longer glycosylated. 29076618 9 4 theme nonulosonic 1488:1498 arg1 transfer 1506:1513 nonulosonic acids transfer 1488:1513 nonulosonic acids transfer onto flagellins 1488:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 4 5 theme many 505:508 arg1 strains 521:527 many pathogenic strains 505:527 many pathogenic strains 505:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 2 6 theme various 111:117 arg1 bacteria 133:140 various Gram-negative bacteria 111:140 various Gram-negative bacteria 111:140 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 9 7 theme family 1469:1474 arg1 member 1423:1428 a member 1421:1428 a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1421:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 6 8 theme gene 882:885 arg1 amb0685 887:893 the gene amb0685 878:893 the gene amb0685 coding for Maf 878:908 Deletion of the gene amb0685 coding for Maf produced a nonflagellated bacterium where the flagellin was still produced but no longer glycosylated. 29076618 5 9 from organism 768:775 arg1 AMB-1 806:810 Magnetospirillum magneticum AMB-1 778:810 Magnetospirillum magneticum AMB-1 778:810 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 9 from organism 768:775 arg1 Maf 749:751 Maf 749:751 Maf from our model organism 749:775 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 9 from organism 768:775 arg1 role 741:744 the role 737:744 the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum 737:863 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 8 10 theme pseudaminic 1236:1246 arg1 structure 1211:1219 the nonulosonic carbohydrate structure 1182:1219 the nonulosonic carbohydrate structure transferred 1182:1231 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 8 10 theme pseudaminic 1236:1246 arg1 acid 1248:1251 pseudaminic acid 1236:1251 pseudaminic acid 1236:1251 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 5 11 from role 741:744 arg1 glycosylation 820:832 glycosylation 820:832 glycosylation 820:832 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 11 from role 741:744 arg1 organism 768:775 our model organism 758:775 our model organism 758:775 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 11 from role 741:744 arg1 formation 838:846 formation 838:846 formation 838:846 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 12 theme model 762:766 arg1 organism 768:775 our model organism 758:775 our model organism 758:775 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 2 13 theme diverse 161:167 arg1 structures 176:185 diverse glycan structures 161:185 diverse glycan structures 161:185 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 9 14 from importance 1305:1314 arg1 formation 1340:1348 the formation 1336:1348 the formation of the bacterial flagellum 1336:1375 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 8 15 theme nonulosonic 1186:1196 arg1 structure 1211:1219 the nonulosonic carbohydrate structure 1182:1219 the nonulosonic carbohydrate structure transferred 1182:1231 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 8 15 theme nonulosonic 1186:1196 arg1 acid 1248:1251 pseudaminic acid 1236:1251 pseudaminic acid 1236:1251 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 4 16 theme pathogenic 510:519 arg1 strains 521:527 many pathogenic strains 505:527 many pathogenic strains 505:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 5 17 dep glycosylation 820:832 arg1 the 816:818 the 816:818 the 816:818 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 1 18 theme bacterial 65:73 arg1 formation 84:92 bacterial flagella formation 65:92 bacterial flagella formation 65:92 The glycosyltransferase Maf is involved in bacterial flagella formation. 29076618 8 19 theme close 1263:1267 arg1 derivative 1269:1278 a very close derivative 1256:1278 a very close derivative 1256:1278 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 4 20 from sugars 611:616 arg1 flagellins 625:634 the flagellins 621:634 the flagellins 621:634 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 9 21 theme acids 1500:1504 arg1 transfer 1506:1513 nonulosonic acids transfer 1488:1513 nonulosonic acids transfer onto flagellins 1488:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 3 22 theme nonulosonic 263:273 arg1 pathways 294:301 nonulosonic sugar biosynthesis pathways 263:301 nonulosonic sugar biosynthesis pathways 263:301 Although nonulosonic sugar biosynthesis pathways have been dissected in various pathogens, the enzymes transferring the sugars onto flagellin are still poorly characterized. 29076618 6 23 gly glycosylated 999:1010 arg1 flagellin 956:964 the flagellin 952:964 the flagellin 952:964 Deletion of the gene amb0685 coding for Maf produced a nonflagellated bacterium where the flagellin was still produced but no longer glycosylated. 29076618 1 24 theme flagella 75:82 arg1 formation 84:92 bacterial flagella formation 65:92 bacterial flagella formation 65:92 The glycosyltransferase Maf is involved in bacterial flagella formation. 29076618 9 25 theme new 1435:1437 arg1 family 1469:1474 a new bacterial glycosyltransferase family 1433:1474 a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1433:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 9 26 theme structural 1400:1409 arg1 model 1411:1415 the first structural model 1390:1415 the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1390:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 7 27 theme Campylobacter 1129:1141 arg1 jejuni 1143:1148 Campylobacter jejuni 1129:1148 Campylobacter jejuni 1129:1148 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 8 28 theme carbohydrate 1198:1209 arg1 structure 1211:1219 the nonulosonic carbohydrate structure 1182:1219 the nonulosonic carbohydrate structure transferred 1182:1231 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 8 28 theme carbohydrate 1198:1209 arg1 acid 1248:1251 pseudaminic acid 1236:1251 pseudaminic acid 1236:1251 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 5 29 theme Maf 749:751 arg1 role 741:744 the role 737:744 the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum 737:863 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 2 30 theme related 219:225 arg1 acids 213:217 nonulosonic acids 201:217 them nonulosonic acids related to the sialic acid family 196:251 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 4 31 theme bacterial 701:709 arg1 motility 711:718 bacterial motility 701:718 bacterial motility 701:718 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 7 32 from jejuni 1143:1148 arg1 sialyltransferases 1105:1122 sialyltransferases 1105:1122 sialyltransferases from Campylobacter jejuni 1105:1148 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 2 33 with decorated 146:154 arg1 structures 176:185 diverse glycan structures 161:185 diverse glycan structures 161:185 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 5 34 theme flagellum 855:863 arg1 glycosylation 820:832 glycosylation 820:832 glycosylation 820:832 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 34 theme flagellum 855:863 arg1 formation 838:846 formation 838:846 formation 838:846 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 4 35 theme nonulosonic 599:609 arg1 sugars 611:616 specific nonulosonic sugars 590:616 specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility 590:718 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 36 theme associated 470:479 arg1 Mafs 490:493 Mafs 490:493 Mafs 490:493 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 36 theme associated 470:479 arg1 factors 481:487 associated factors 470:487 associated factors (Mafs) found in many pathogenic strains 470:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 37 theme Maf 657:659 arg1 function 661:668 Maf function 657:668 Maf function 657:668 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 4 38 theme specific 590:597 arg1 sugars 611:616 specific nonulosonic sugars 590:616 specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility 590:718 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 3 39 theme sugar 275:279 arg1 pathways 294:301 nonulosonic sugar biosynthesis pathways 263:301 nonulosonic sugar biosynthesis pathways 263:301 Although nonulosonic sugar biosynthesis pathways have been dissected in various pathogens, the enzymes transferring the sugars onto flagellin are still poorly characterized. 29076618 2 40 theme acid 241:244 arg1 family 246:251 the sialic acid family 230:251 the sialic acid family 230:251 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 4 41 theme flagellin 673:681 arg1 glycosylation 683:695 flagellin glycosylation 673:695 flagellin glycosylation 673:695 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 3 42 theme biosynthesis 281:292 arg1 pathways 294:301 nonulosonic sugar biosynthesis pathways 263:301 nonulosonic sugar biosynthesis pathways 263:301 Although nonulosonic sugar biosynthesis pathways have been dissected in various pathogens, the enzymes transferring the sugars onto flagellin are still poorly characterized. 29076618 6 43 theme amb0685 887:893 arg1 Deletion 866:873 Deletion 866:873 Deletion of the gene amb0685 coding for Maf 866:908 Deletion of the gene amb0685 coding for Maf produced a nonflagellated bacterium where the flagellin was still produced but no longer glycosylated. 29076618 2 44 theme sialic 234:239 arg1 family 246:251 the sialic acid family 230:251 the sialic acid family 230:251 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 9 45 theme bacterial 1439:1447 arg1 family 1469:1474 a new bacterial glycosyltransferase family 1433:1474 a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1433:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 7 46 theme Maf 1078:1080 arg1 Maf 1078:1080 Maf 1078:1080 Maf 1078:1080 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 7 46 theme Maf 1078:1080 arg1 domain 1068:1073 the central domain 1056:1073 the central domain of Maf 1056:1080 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 2 47 theme glycan 169:174 arg1 structures 176:185 diverse glycan structures 161:185 diverse glycan structures 161:185 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 7 48 theme central 1060:1066 arg1 Maf 1078:1080 Maf 1078:1080 Maf 1078:1080 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 7 48 theme central 1060:1066 arg1 domain 1068:1073 the central domain 1056:1073 the central domain of Maf 1056:1080 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 9 49 theme flagellum 1367:1375 arg1 formation 1340:1348 the formation 1336:1348 the formation of the bacterial flagellum 1336:1375 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 4 50 theme nonflagellated 558:571 arg1 bacteria 573:580 nonflagellated bacteria 558:580 nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility 558:718 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 3 51 theme various 326:332 arg1 pathogens 334:342 various pathogens 326:342 various pathogens 326:342 Although nonulosonic sugar biosynthesis pathways have been dissected in various pathogens, the enzymes transferring the sugars onto flagellin are still poorly characterized. 29076618 4 52 theme genes 444:448 arg1 deletion 432:439 The deletion 428:439 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains 428:527 The deletion of genes coding for motility associated factors (Mafs) found in many pathogenic strains systematically gives rise to nonflagellated bacteria lacking specific nonulosonic sugars on the flagellins, therefore, relating Maf function to flagellin glycosylation and bacterial motility. 29076618 9 53 theme glycosylation 1319:1331 arg1 importance 1305:1314 the importance 1301:1314 the importance of glycosylation in the formation of the bacterial flagellum 1301:1375 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 9 54 theme bacterial 1357:1365 arg1 flagellum 1367:1375 the bacterial flagellum 1353:1375 the bacterial flagellum 1353:1375 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 9 55 theme first 1394:1398 arg1 model 1411:1415 the first structural model 1390:1415 the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1390:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 8 56 theme Glycan 1151:1156 arg1 analysis 1158:1165 Glycan analysis 1151:1165 Glycan analysis 1151:1165 Glycan analysis suggested that the nonulosonic carbohydrate structure transferred is pseudaminic acid or a very close derivative. 29076618 9 57 theme glycosyltransferase 1449:1467 arg1 family 1469:1474 a new bacterial glycosyltransferase family 1433:1474 a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins 1433:1529 This work describes the importance of glycosylation in the formation of the bacterial flagellum and provides the first structural model for a member of a new bacterial glycosyltransferase family involved in nonulosonic acids transfer onto flagellins. 29076618 2 58 theme nonulosonic 201:211 arg1 acids 213:217 nonulosonic acids 201:217 them nonulosonic acids related to the sialic acid family 196:251 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 1 59 theme glycosyltransferase 26:44 arg1 Maf 46:48 The glycosyltransferase Maf 22:48 The glycosyltransferase Maf 22:48 The glycosyltransferase Maf is involved in bacterial flagella formation. 29076618 7 60 theme structure 1023:1031 arg1 analysis 1033:1040 Our X-ray structure analysis 1013:1040 Our X-ray structure analysis 1013:1040 Our X-ray structure analysis revealed that the central domain of Maf exhibits similarity to sialyltransferases from Campylobacter jejuni. 29076618 2 61 dep them 196:199 arg1 acids 213:217 nonulosonic acids 201:217 them nonulosonic acids related to the sialic acid family 196:251 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 5 62 theme Magnetospirillum 778:793 arg1 Maf 749:751 Maf 749:751 Maf from our model organism 749:775 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 62 theme Magnetospirillum 778:793 arg1 AMB-1 806:810 Magnetospirillum magneticum AMB-1 778:810 Magnetospirillum magneticum AMB-1 778:810 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 63 theme magneticum 795:804 arg1 Maf 749:751 Maf 749:751 Maf from our model organism 749:775 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 5 63 theme magneticum 795:804 arg1 AMB-1 806:810 Magnetospirillum magneticum AMB-1 778:810 Magnetospirillum magneticum AMB-1 778:810 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 29076618 2 64 theme bacteria 133:140 arg1 decorated 146:154 decorated 146:154 decorated 146:154 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 2 64 theme bacteria 133:140 arg1 flagella 99:106 The flagella 95:106 The flagella of various Gram-negative bacteria 95:140 The flagella of various Gram-negative bacteria are decorated with diverse glycan structures, amongst them nonulosonic acids related to the sialic acid family. 29076618 5 65 gly glycosylation 820:832 arg1 flagellum 855:863 the flagellum 851:863 the flagellum 851:863 We investigated the role of Maf from our model organism, Magnetospirillum magneticum AMB-1, in the glycosylation and formation of the flagellum. 25976627 1 0 theme cell 166:169 arg1 activity 192:199 whole cell transgalactosylating activity 160:199 whole cell transgalactosylating activity 160:199 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 0 1 theme NCIM 104:107 arg1 3551 109:112 Kluyveromyces marxianus NCIM 3551 80:112 Kluyveromyces marxianus NCIM 3551 80:112 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 25976627 1 2 theme transgalactosylating 171:190 arg1 activity 192:199 whole cell transgalactosylating activity 160:199 whole cell transgalactosylating activity 160:199 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 4 3 theme 24g/L/h 621:627 arg1 % 599:599 80% 597:599 80% 597:599 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 3 theme 24g/L/h 621:627 arg1 24g/L/h 621:627 24g/L/h 621:627 24g/L/h 621:627 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 3 theme 24g/L/h 621:627 arg1 productivity 605:616 productivity 605:616 productivity 605:616 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 4 from units 746:750 arg1 conditions 660:669 the optimum conditions 648:669 the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units 648:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 5 theme enzyme 739:744 arg1 units 746:750 enzyme units 739:750 enzyme units 739:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 6 theme temperature 710:720 arg1 40°C 722:725 temperature 40°C 710:725 temperature 40°C 710:725 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 7 from concentration 682:694 arg1 conditions 660:669 the optimum conditions 648:669 the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units 648:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 8 theme % 599:599 arg1 rate 589:592 A lactose conversion rate 568:592 A lactose conversion rate of 80% and productivity of 24g/L/h 568:627 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 3 9 theme % 460:460 arg1 yield 449:453 A maximum GOS yield 435:453 A maximum GOS yield of 36% (w/w) of total sugars 435:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 0 10 theme galacto-oligosaccharides 49:72 arg1 synthesis 36:44 synthesis 36:44 synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551 36:112 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 25976627 4 11 theme productivity 605:616 arg1 rate 589:592 A lactose conversion rate 568:592 A lactose conversion rate of 80% and productivity of 24g/L/h 568:627 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 2 12 theme GOS 430:432 arg1 synthesis 417:425 synthesis 417:425 synthesis of GOS 417:432 Cell permeabilization lead to an efficient bioconversion by β-galactosidase resulting in synthesis of GOS. 25976627 4 13 theme conversion 578:587 arg1 rate 589:592 A lactose conversion rate 568:592 A lactose conversion rate of 80% and productivity of 24g/L/h 568:627 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 14 theme lactose 570:576 arg1 rate 589:592 A lactose conversion rate 568:592 A lactose conversion rate of 80% and productivity of 24g/L/h 568:627 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 1 15 theme Kluyveromyces 202:214 arg1 3551 231:234 Kluyveromyces marxianus NCIM 3551 202:234 Kluyveromyces marxianus NCIM 3551 202:234 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 1 16 theme galacto-oligosaccharides 296:319 arg1 production 282:291 production 282:291 production of galacto-oligosaccharides (GOS) 282:325 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 1 17 theme marxianus 216:224 arg1 3551 231:234 Kluyveromyces marxianus NCIM 3551 202:234 Kluyveromyces marxianus NCIM 3551 202:234 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 4 18 from 40°C 722:725 arg1 conditions 660:669 the optimum conditions 648:669 the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units 648:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 2 19 theme Cell 328:331 arg1 permeabilization 333:348 Cell permeabilization 328:348 Cell permeabilization 328:348 Cell permeabilization lead to an efficient bioconversion by β-galactosidase resulting in synthesis of GOS. 25976627 1 20 theme NCIM 226:229 arg1 3551 231:234 Kluyveromyces marxianus NCIM 3551 202:234 Kluyveromyces marxianus NCIM 3551 202:234 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 0 21 theme lactose 24:30 arg1 Transgalactosylation 0:19 Transgalactosylation 0:19 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 0:113 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 25976627 4 22 theme % 701:701 arg1 pH 728:729 pH 6.5 728:733 pH 6.5 728:733 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 22 theme % 701:701 arg1 40°C 722:725 temperature 40°C 710:725 temperature 40°C 710:725 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 22 theme % 701:701 arg1 units 746:750 enzyme units 739:750 enzyme units 739:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 22 theme % 701:701 arg1 concentration 682:694 lactose concentration 674:694 lactose concentration of 20% (w/v) 674:707 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 3 23 theme total 471:475 arg1 sugars 477:482 total sugars 471:482 total sugars 471:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 3 24 theme GOS 445:447 arg1 yield 449:453 A maximum GOS yield 435:453 A maximum GOS yield of 36% (w/w) of total sugars 435:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 2 25 theme efficient 361:369 arg1 bioconversion 371:383 an efficient bioconversion 358:383 an efficient bioconversion 358:383 Cell permeabilization lead to an efficient bioconversion by β-galactosidase resulting in synthesis of GOS. 25976627 4 26 theme lactose 674:680 arg1 concentration 682:694 lactose concentration 674:694 lactose concentration of 20% (w/v) 674:707 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 4 27 theme optimum 652:658 arg1 conditions 660:669 the optimum conditions 648:669 the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units 648:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 5 28 theme reaction 827:834 arg1 mixture 836:842 the reaction mixture 823:842 the reaction mixture 823:842 Tetrasaccharides were the main component of the reaction mixture. 25976627 3 29 theme maximum 437:443 arg1 yield 449:453 A maximum GOS yield 435:453 A maximum GOS yield of 36% (w/w) of total sugars 435:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 6 30 theme mass 917:920 arg1 spectrometry 922:933 mass spectrometry 917:933 mass spectrometry 917:933 The products were quantitated by HPLC and structurally characterized by mass spectrometry. 25976627 1 31 theme yeast 133:137 arg1 strains 139:145 yeast strains 133:145 yeast strains screened for whole cell transgalactosylating activity 133:199 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 1 32 theme strains 139:145 arg1 number 123:128 a number 121:128 a number of yeast strains screened for whole cell transgalactosylating activity 121:199 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 4 33 from pH 728:729 arg1 conditions 660:669 the optimum conditions 648:669 the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units 648:750 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 3 34 theme sugars 477:482 arg1 % 460:460 36% 458:460 36% (w/w) of total sugars 458:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 3 34 theme sugars 477:482 arg1 sugars 477:482 total sugars 471:482 total sugars 471:482 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 3 34 theme sugars 477:482 arg1 w/w 463:465 w/w 463:465 w/w 463:465 A maximum GOS yield of 36% (w/w) of total sugars was achieved and the products consisted of tri- and tetra-galacto-oligosaccharides. 25976627 0 35 theme marxianus 94:102 arg1 3551 109:112 Kluyveromyces marxianus NCIM 3551 80:112 Kluyveromyces marxianus NCIM 3551 80:112 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 25976627 4 36 theme time 773:776 arg1 3h 758:759 3h 758:759 3h of reaction time 758:776 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 5 37 theme main 805:808 arg1 Tetrasaccharides 779:794 Tetrasaccharides 779:794 Tetrasaccharides 779:794 Tetrasaccharides were the main component of the reaction mixture. 25976627 5 37 theme main 805:808 arg1 component 810:818 the main component 801:818 the main component of the reaction mixture 801:842 Tetrasaccharides were the main component of the reaction mixture. 25976627 5 38 theme mixture 836:842 arg1 Tetrasaccharides 779:794 Tetrasaccharides 779:794 Tetrasaccharides 779:794 Tetrasaccharides were the main component of the reaction mixture. 25976627 5 38 theme mixture 836:842 arg1 component 810:818 the main component 801:818 the main component of the reaction mixture 801:842 Tetrasaccharides were the main component of the reaction mixture. 25976627 0 39 theme Kluyveromyces 80:92 arg1 3551 109:112 Kluyveromyces marxianus NCIM 3551 80:112 Kluyveromyces marxianus NCIM 3551 80:112 Transgalactosylation of lactose for synthesis of galacto-oligosaccharides using Kluyveromyces marxianus NCIM 3551. 25976627 4 40 theme reaction 764:771 arg1 time 773:776 reaction time 764:776 reaction time 764:776 A lactose conversion rate of 80% and productivity of 24g/L/h was obtained under the optimum conditions at lactose concentration of 20% (w/v), temperature 40°C, pH 6.5 and enzyme units after 3h of reaction time. 25976627 1 41 theme whole 160:164 arg1 activity 192:199 whole cell transgalactosylating activity 160:199 whole cell transgalactosylating activity 160:199 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 25976627 1 42 theme suitable 257:264 arg1 biocatalyst 266:276 most suitable biocatalyst 252:276 most suitable biocatalyst 252:276 Among a number of yeast strains screened for whole cell transgalactosylating activity, Kluyveromyces marxianus NCIM 3551 was found to be most suitable biocatalyst for production of galacto-oligosaccharides (GOS). 28734224 5 0 theme glucosyl 1098:1105 arg1 α1→3 1116:1119 α1→3 1116:1119 α1→3 1116:1119 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 5 0 theme glucosyl 1098:1105 arg1 residue 1107:1113 a glucosyl residue 1096:1113 a glucosyl residue 1096:1113 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 6 1 theme glucosyl 1230:1237 arg1 residue 1239:1245 a glucosyl residue 1228:1245 a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose 1228:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 3 2 theme polymerization 749:762 arg1 degree 739:744 a degree 737:744 a degree of polymerization (DP) of 3-4 737:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 1 3 theme products 294:301 arg1 range 244:248 their wide range 233:248 their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential 233:354 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 2 4 theme other 517:521 arg1 molecules 523:531 other molecules 517:531 other molecules 517:531 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 4 theme other 517:521 arg1 substrates 545:554 acceptor substrates 536:554 acceptor substrates 536:554 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 3 5 theme lactose 717:723 arg1 products 725:732 at least 5 glucosylated lactose products 693:732 at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4 693:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 1 6 from bacteria 183:190 arg1 enzymes 158:164 Glucansucrase enzymes 144:164 Glucansucrase enzymes from lactic acid bacteria 144:190 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 2 7 theme Lactobacillus 393:405 arg1 strains 415:421 Lactobacillus reuteri strains 393:421 Lactobacillus reuteri strains 393:421 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 0 8 theme Gtf180 114:119 arg1 enzymes 135:141 Gtf180 glucansucrase enzymes 114:141 Gtf180 glucansucrase enzymes 114:141 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 3 9 theme products 725:732 arg1 synthesis 680:688 synthesis 680:688 synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4 680:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 1 10 contain have 331:334 arg1 some 317:320 some 317:320 some 317:320 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 1 10 contain have 331:334 arg2 potential 346:354 prebiotic potential 336:354 prebiotic potential 336:354 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 1 10 contain have 331:334 arg1 products 294:301 gluco-oligosaccharide and polysaccharide products 253:301 gluco-oligosaccharide and polysaccharide products 253:301 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 5 11 theme non-reducing 1147:1158 arg1 unit 1170:1173 the non-reducing galactose unit 1143:1173 the non-reducing galactose unit of lactose 1143:1184 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 7 12 link derived 1316:1322 arg1 compounds 1379:1387 interesting prebiotic compounds 1357:1387 interesting prebiotic compounds 1357:1387 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 7 12 link derived 1316:1322 arg1 oligosaccharides 1324:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 2 13 with α-glucans 457:465 arg1 types 482:486 different types 472:486 different types of linkages 472:498 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 14 dep Lactobacillus 393:405 arg1 reuteri 407:413 reuteri 407:413 reuteri 407:413 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 5 15 theme galactose 1160:1168 arg1 unit 1170:1173 the non-reducing galactose unit 1143:1173 the non-reducing galactose unit of lactose 1143:1184 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 6 16 theme glucose 1277:1283 arg1 unit 1285:1288 the reducing glucose unit 1264:1288 the reducing glucose unit of lactose 1264:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 7 17 theme interesting 1357:1367 arg1 compounds 1379:1387 interesting prebiotic compounds 1357:1387 interesting prebiotic compounds 1357:1387 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 7 17 theme interesting 1357:1367 arg1 oligosaccharides 1324:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 5 18 theme other 968:972 arg1 bonds 962:966 glycosidic bonds 951:966 glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN 951:1051 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 4 19 dep lactose-based 827:839 arg1 larger 820:825 larger 820:825 larger 820:825 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 0 20 theme glucansucrase 121:133 arg1 enzymes 135:141 Gtf180 glucansucrase enzymes 114:141 Gtf180 glucansucrase enzymes 114:141 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 3 21 theme degree 739:744 arg1 products 725:732 at least 5 glucosylated lactose products 693:732 at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4 693:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 6 22 theme reducing 1268:1275 arg1 unit 1285:1288 the reducing glucose unit 1264:1288 the reducing glucose unit of lactose 1264:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 2 23 theme linkages 491:498 arg1 types 482:486 different types 472:486 different types of linkages 472:498 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 1 24 theme strong 206:211 arg1 interest 213:220 strong interest 206:220 strong interest 206:220 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 6 25 theme lactose 1293:1299 arg1 unit 1285:1288 the reducing glucose unit 1264:1288 the reducing glucose unit of lactose 1264:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 2 26 theme acceptor 536:543 arg1 molecules 523:531 other molecules 517:531 other molecules 517:531 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 26 theme acceptor 536:543 arg1 substrates 545:554 acceptor substrates 536:554 acceptor substrates 536:554 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 3 27 theme truncated 605:613 arg1 versions 615:622 N-terminally truncated versions 592:622 N-terminally truncated versions of 592:625 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 4 28 dep produced 811:818 arg1 DP9 865:867 DP9 865:867 DP9 865:867 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 0 29 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of glucosylated lactose derivatives 0:62 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 3 30 dep enzymes 634:640 arg1 versions 615:622 N-terminally truncated versions 592:622 N-terminally truncated versions of 592:625 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 0 31 theme glucosylated 31:42 arg1 derivatives 52:62 glucosylated lactose derivatives 31:62 glucosylated lactose derivatives 31:62 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 3 32 theme 3-4 772:774 arg1 DP 765:766 DP 765:766 DP 765:766 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 3 32 theme 3-4 772:774 arg1 polymerization 749:762 polymerization 749:762 polymerization (DP) of 3-4 749:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 1 33 from sucrose 308:314 arg1 products 294:301 gluco-oligosaccharide and polysaccharide products 253:301 gluco-oligosaccharide and polysaccharide products 253:301 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 1 33 from sucrose 308:314 arg1 range 244:248 their wide range 233:248 their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential 233:354 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 2 34 from strains 415:421 arg1 Gtf180 381:386 Gtf180 381:386 Gtf180 381:386 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 34 from strains 415:421 arg1 GtfA 372:375 GtfA 372:375 GtfA 372:375 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 34 from strains 415:421 arg1 Glucansucrases 357:370 Glucansucrases GtfA and Gtf180 357:386 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains 357:421 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 0 35 dep Lactobacillus 83:95 arg1 reuteri 97:103 reuteri 97:103 reuteri 97:103 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 2 36 theme different 472:480 arg1 types 482:486 different types 472:486 different types of linkages 472:498 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 4 37 dep DP9 865:867 arg1 to 862:863 to 862:863 to 862:863 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 5 38 dep products 927:934 arg1 DP3-4 936:940 DP3-4 936:940 DP3-4 936:940 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 5 39 theme lactose 919:925 arg1 products 927:934 the glucosylated lactose products DP3-4 902:940 the glucosylated lactose products DP3-4 902:940 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 0 40 theme derivatives 52:62 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of glucosylated lactose derivatives 0:62 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 6 41 theme -linked 1253:1259 arg1 residue 1239:1245 a glucosyl residue 1228:1245 a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose 1228:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 5 42 theme glucosylated 906:917 arg1 products 927:934 the glucosylated lactose products DP3-4 902:940 the glucosylated lactose products DP3-4 902:940 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 1 43 theme wide 239:242 arg1 range 244:248 their wide range 233:248 their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential 233:354 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 0 44 theme lactose 44:50 arg1 derivatives 52:62 glucosylated lactose derivatives 31:62 glucosylated lactose derivatives 31:62 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 1 45 theme prebiotic 336:344 arg1 potential 346:354 prebiotic potential 336:354 prebiotic potential 336:354 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 3 46 theme enzymes 634:640 arg1 incubation 577:586 incubation 577:586 incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose 577:666 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 5 47 theme Structural 871:880 arg1 characterization 882:897 Structural characterization 871:897 Structural characterization of the glucosylated lactose products DP3-4 871:940 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 3 48 with incubation 577:586 arg1 lactose 647:653 lactose 647:653 lactose 647:653 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 3 48 with incubation 577:586 arg1 sucrose 660:666 sucrose 660:666 sucrose 660:666 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 1 49 from range 244:248 arg1 sucrose 308:314 sucrose 308:314 sucrose 308:314 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 5 50 theme glycosidic 951:960 arg1 bonds 962:966 glycosidic bonds 951:966 glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN 951:1051 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 1 51 theme Glucansucrase 144:156 arg1 enzymes 158:164 Glucansucrase enzymes 144:164 Glucansucrase enzymes from lactic acid bacteria 144:190 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 6 52 link -linked 1253:1259 arg1 residue 1239:1245 a glucosyl residue 1228:1245 a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose 1228:1299 Both enzymes also were able to introduce a glucosyl residue (α1→2)-linked to the reducing glucose unit of lactose. 28734224 7 53 theme prebiotic 1369:1377 arg1 compounds 1379:1387 interesting prebiotic compounds 1357:1387 interesting prebiotic compounds 1357:1387 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 7 53 theme prebiotic 1369:1377 arg1 oligosaccharides 1324:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 3 54 theme glucosylated 704:715 arg1 products 725:732 at least 5 glucosylated lactose products 693:732 at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4 693:774 Here we report that incubation of (N-terminally truncated versions of) these enzymes with lactose plus sucrose resulted in synthesis of at least 5 glucosylated lactose products of a degree of polymerization (DP) of 3-4. 28734224 2 55 dep Glucansucrases 357:370 arg1 Gtf180 381:386 Gtf180 381:386 Gtf180 381:386 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 55 dep Glucansucrases 357:370 arg1 GtfA 372:375 GtfA 372:375 GtfA 372:375 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 2 55 dep Glucansucrases 357:370 arg1 Glucansucrases 357:370 Glucansucrases GtfA and Gtf180 357:386 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains 357:421 Glucansucrases GtfA and Gtf180 from Lactobacillus reuteri strains are known to convert sucrose into α-glucans with different types of linkages, but also to use other molecules as acceptor substrates. 28734224 7 56 theme derived 1316:1322 arg1 compounds 1379:1387 interesting prebiotic compounds 1357:1387 interesting prebiotic compounds 1357:1387 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 7 56 theme derived 1316:1322 arg1 oligosaccharides 1324:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides 1302:1339 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 4 57 theme glucansucrase 782:794 arg1 Gtf180-ΔN 796:804 Only glucansucrase Gtf180-ΔN 777:804 Only glucansucrase Gtf180-ΔN 777:804 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 4 58 theme Only 777:780 arg1 Gtf180-ΔN 796:804 Only glucansucrase Gtf180-ΔN 777:804 Only glucansucrase Gtf180-ΔN 777:804 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 5 59 theme products 927:934 arg1 characterization 882:897 Structural characterization 871:897 Structural characterization of the glucosylated lactose products DP3-4 871:940 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 1 60 theme gluco-oligosaccharide 253:273 arg1 products 294:301 gluco-oligosaccharide and polysaccharide products 253:301 gluco-oligosaccharide and polysaccharide products 253:301 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 0 61 theme Lactobacillus 83:95 arg1 GtfA 105:108 the Lactobacillus reuteri GtfA 79:108 the Lactobacillus reuteri GtfA 79:108 Structural characterization of glucosylated lactose derivatives synthesized by the Lactobacillus reuteri GtfA and Gtf180 glucansucrase enzymes. 28734224 4 62 theme lactose-based 827:839 arg1 oligosaccharides 841:856 larger lactose-based oligosaccharides 820:856 larger lactose-based oligosaccharides 820:856 Only glucansucrase Gtf180-ΔN also produced larger lactose-based oligosaccharides (up to DP9). 28734224 1 63 theme lactic 171:176 arg1 bacteria 183:190 lactic acid bacteria 171:190 lactic acid bacteria 171:190 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 5 64 theme lactose 1178:1184 arg1 unit 1170:1173 the non-reducing galactose unit 1143:1173 the non-reducing galactose unit of lactose 1143:1184 Structural characterization of the glucosylated lactose products DP3-4 revealed glycosidic bonds other than (α1→4)/(α1→6) typical for GtfA-ΔN and (α1→3)/(α1→6) typical for Gtf180-ΔN: Both GtfA-ΔN and Gtf180-ΔN now introduced a glucosyl residue (α1→3)- or (α1→4)-linked to the non-reducing galactose unit of lactose. 28734224 7 65 dep derived 1316:1322 arg1 lactose 1308:1314 lactose 1308:1314 lactose 1308:1314 These lactose derived oligosaccharides potentially are interesting prebiotic compounds. 28734224 1 66 theme acid 178:181 arg1 bacteria 183:190 lactic acid bacteria 171:190 lactic acid bacteria 171:190 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 28734224 1 67 theme polysaccharide 279:292 arg1 products 294:301 gluco-oligosaccharide and polysaccharide products 253:301 gluco-oligosaccharide and polysaccharide products 253:301 Glucansucrase enzymes from lactic acid bacteria are receiving strong interest because of their wide range of gluco-oligosaccharide and polysaccharide products from sucrose, some of which have prebiotic potential. 30023556 0 0 theme MUC1 99:102 arg1 Library 117:123 MUC1 Glycopeptide Library 99:123 MUC1 Glycopeptide Library 99:123 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 8 1 theme strong 1178:1183 arg1 binding 1185:1191 a strong binding 1176:1191 a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope 1176:1270 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 10 2 theme predesigned 1499:1509 arg1 specificities 1520:1532 predesigned O-glycan specificities 1499:1532 predesigned O-glycan specificities 1499:1532 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 8 3 theme monovalent 1253:1262 arg1 epitope 1264:1270 a monovalent epitope 1251:1270 a monovalent epitope 1251:1270 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 10 4 theme antibodies 1483:1492 arg1 development 1462:1472 the development 1458:1472 the development of novel antibodies with predesigned O-glycan specificities 1458:1532 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 6 5 dep position 990:997 arg1 dST 1015:1017 dST 1015:1017 dST 1015:1017 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 5 dep position 990:997 arg1 STn 1000:1002 STn 1000:1002 STn 1000:1002 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 5 dep position 990:997 arg1 26ST 1005:1008 26ST 1005:1008 26ST 1005:1008 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 6 theme unsubstituted 878:890 arg1 position 896:903 an unsubstituted O-6 position 875:903 an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST) 875:943 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 7 theme same 985:988 arg1 position 990:997 the same position 981:997 the same position (STn, 26ST, and dST) 981:1018 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 7 8 theme core 1065:1068 arg1 O-glycans 1072:1080 core 2 O-glycans 1065:1080 core 2 O-glycans 1065:1080 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 7 9 contain have 1087:1090 arg1 glycopeptides 1046:1058 glycopeptides 1046:1058 glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue 1046:1139 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 7 9 contain have 1087:1090 arg2 GlcNAc 1092:1097 GlcNAc 1092:1097 GlcNAc 1092:1097 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 2 10 theme low 319:321 arg1 affinity 323:330 low affinity 319:330 low affinity 319:330 However, most of them show low specificities toward O-glycan structures and/or low affinity toward a monovalent epitope. 30023556 9 11 theme MUC1 1360:1363 arg1 structures 1374:1383 MUC1 O-glycan structures 1360:1383 MUC1 O-glycan structures 1360:1383 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 0 12 theme Glycopeptide 104:115 arg1 Library 117:123 MUC1 Glycopeptide Library 99:123 MUC1 Glycopeptide Library 99:123 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 8 13 gly glycopeptide 1233:1244 arg2 glycopeptide 1233:1244 20-mer glycopeptide 1226:1244 not only native MUC1 but also 20-mer glycopeptide 1196:1244 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 10 14 gly glycopeptide 1421:1432 arg2 glycopeptide 1421:1432 glycopeptide libraries 1421:1442 glycopeptide libraries 1421:1442 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 2 15 theme monovalent 341:350 arg1 epitope 352:358 a monovalent epitope 339:358 a monovalent epitope 339:358 However, most of them show low specificities toward O-glycan structures and/or low affinity toward a monovalent epitope. 30023556 7 16 theme GalNAc 1126:1131 arg1 residue 1133:1139 the GalNAc residue 1122:1139 the GalNAc residue 1122:1139 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 3 17 theme designed 490:497 arg1 specificities 512:524 designed carbohydrate specificities 490:524 designed carbohydrate specificities 490:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 2 18 theme O-glycan 292:299 arg1 structures 301:310 O-glycan structures 292:310 O-glycan structures 292:310 However, most of them show low specificities toward O-glycan structures and/or low affinity toward a monovalent epitope. 30023556 4 19 theme cell 707:710 arg1 lines 712:716 various cell lines 699:716 various cell lines 699:716 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 1 20 theme O-glycan 186:193 arg1 structures 200:209 O-glycan core structures 186:209 O-glycan core structures 186:209 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures have already been developed. 30023556 4 21 theme different 610:618 arg1 features 620:627 quite different features 604:627 quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines 604:716 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 6 22 theme GalNAc 912:917 arg1 residue 919:925 the GalNAc residue 908:925 the GalNAc residue (Tn, T, and 23ST) 908:943 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 3 23 gly glycopeptide 390:401 arg2 glycopeptide 390:401 an MUC1 glycopeptide library 382:409 an MUC1 glycopeptide library 382:409 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 10 24 theme glycopeptide 1421:1432 arg1 libraries 1434:1442 glycopeptide libraries 1421:1442 glycopeptide libraries 1421:1442 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 1 25 theme core 195:198 arg1 structures 200:209 O-glycan core structures 186:209 O-glycan core structures 186:209 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures have already been developed. 30023556 3 26 theme glycopeptide 390:401 arg1 library 403:409 an MUC1 glycopeptide library 382:409 an MUC1 glycopeptide library 382:409 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 10 27 theme O-glycan 1511:1518 arg1 specificities 1520:1532 predesigned O-glycan specificities 1499:1532 predesigned O-glycan specificities 1499:1532 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 6 28 theme residue 919:925 arg1 position 896:903 an unsubstituted O-6 position 875:903 an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST) 875:943 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 3 29 theme novel 431:435 arg1 1B2 470:472 1B2 470:472 1B2 470:472 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 29 theme novel 431:435 arg1 antibodies 458:467 two novel anti-MUC1 monoclonal antibodies 427:467 two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities 427:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 29 theme novel 431:435 arg1 12D10 478:482 12D10 478:482 12D10 478:482 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 9 30 from studies 1349:1355 arg1 structures 1374:1383 MUC1 O-glycan structures 1360:1383 MUC1 O-glycan structures 1360:1383 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 3 31 theme anti-MUC1 437:445 arg1 1B2 470:472 1B2 470:472 1B2 470:472 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 31 theme anti-MUC1 437:445 arg1 antibodies 458:467 two novel anti-MUC1 monoclonal antibodies 427:467 two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities 427:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 31 theme anti-MUC1 437:445 arg1 12D10 478:482 12D10 478:482 12D10 478:482 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 0 32 theme Anti-MUC1 20:28 arg1 Antibodies 41:50 Novel Anti-MUC1 Monoclonal Antibodies 14:50 Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities 14:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 3 33 theme MUC1 385:388 arg1 library 403:409 an MUC1 glycopeptide library 382:409 an MUC1 glycopeptide library 382:409 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 5 34 theme specific 746:753 arg1 structures 764:773 specific O-glycan structures 746:773 specific O-glycan structures 746:773 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 1 35 theme Numerous 126:133 arg1 antibodies 160:169 Numerous anti-mucin 1 (anti-MUC1) antibodies 126:169 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures 126:209 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures have already been developed. 30023556 9 36 theme O-glycan 1365:1372 arg1 structures 1374:1383 MUC1 O-glycan structures 1360:1383 MUC1 O-glycan structures 1360:1383 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 0 37 theme Antibodies 41:50 arg1 Generation 0:9 Generation 0:9 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities 0:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 9 38 theme anti-MUC1 1279:1287 arg1 antibodies 1289:1298 These anti-MUC1 antibodies 1273:1298 These anti-MUC1 antibodies 1273:1298 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 5 39 theme O-glycan 755:762 arg1 structures 764:773 specific O-glycan structures 746:773 specific O-glycan structures 746:773 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 8 40 theme native 1205:1210 arg1 MUC1 1212:1215 native MUC1 1205:1215 not only native MUC1 but also 20-mer glycopeptide 1196:1244 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 4 41 theme various 699:705 arg1 lines 712:716 various cell lines 699:716 various cell lines 699:716 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 4 42 theme reactivity 676:685 arg1 profiles 687:694 reactivity profiles 676:694 reactivity profiles 676:694 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 0 43 theme Monoclonal 30:39 arg1 Antibodies 41:50 Novel Anti-MUC1 Monoclonal Antibodies 14:50 Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities 14:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 4 44 theme reported 552:559 arg1 antibodies 571:580 previously reported anti-MUC1 antibodies 541:580 previously reported anti-MUC1 antibodies 541:580 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 5 45 dep motif 788:792 arg1 represents 808:817 represents 808:817 represents an O-glycosylation site 808:841 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 6 46 theme O-6 892:894 arg1 position 896:903 an unsubstituted O-6 position 875:903 an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST) 875:943 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 7 47 theme O-6 1106:1108 arg1 position 1110:1117 the O-6 position 1102:1117 the O-6 position of the GalNAc residue 1102:1139 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 5 48 theme * 785:785 arg1 motif 788:792 the PDT*R motif 778:792 the PDT*R motif (the asterisk represents an O-glycosylation site) 778:842 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 0 49 theme Designed 57:64 arg1 Specificities 79:91 Designed Carbohydrate Specificities 57:91 Designed Carbohydrate Specificities 57:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 10 50 theme novel 1477:1481 arg1 antibodies 1483:1492 novel antibodies 1477:1492 novel antibodies with predesigned O-glycan specificities 1477:1532 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 3 51 theme monoclonal 447:456 arg1 1B2 470:472 1B2 470:472 1B2 470:472 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 51 theme monoclonal 447:456 arg1 antibodies 458:467 two novel anti-MUC1 monoclonal antibodies 427:467 two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities 427:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 51 theme monoclonal 447:456 arg1 12D10 478:482 12D10 478:482 12D10 478:482 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 1 52 theme anti-mucin 135:144 arg1 antibodies 160:169 Numerous anti-mucin 1 (anti-MUC1) antibodies 126:169 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures 126:209 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures have already been developed. 30023556 5 53 theme O-glycosylation 822:836 arg1 site 838:841 an O-glycosylation site 819:841 an O-glycosylation site 819:841 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 7 54 theme residue 1133:1139 arg1 position 1110:1117 the O-6 position 1102:1117 the O-6 position of the GalNAc residue 1102:1139 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 3 55 dep antibodies 458:467 arg1 1B2 470:472 1B2 470:472 1B2 470:472 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 55 dep antibodies 458:467 arg1 antibodies 458:467 two novel anti-MUC1 monoclonal antibodies 427:467 two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities 427:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 55 dep antibodies 458:467 arg1 12D10 478:482 12D10 478:482 12D10 478:482 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 3 56 theme carbohydrate 499:510 arg1 specificities 512:524 designed carbohydrate specificities 490:524 designed carbohydrate specificities 490:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 4 57 theme anti-MUC1 561:569 arg1 antibodies 571:580 previously reported anti-MUC1 antibodies 541:580 previously reported anti-MUC1 antibodies 541:580 Compared with previously reported anti-MUC1 antibodies, 1B2 and 12D10 showed quite different features regarding their specificities, affinities, and reactivity profiles to various cell lines. 30023556 10 58 with antibodies 1483:1492 arg1 specificities 1520:1532 predesigned O-glycan specificities 1499:1532 predesigned O-glycan specificities 1499:1532 Furthermore, the strategy of using glycopeptide libraries should enable the development of novel antibodies with predesigned O-glycan specificities. 30023556 7 59 with glycopeptides 1046:1058 arg1 O-glycans 1072:1080 core 2 O-glycans 1065:1080 core 2 O-glycans 1065:1080 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 9 60 theme powerful 1319:1326 arg1 tools 1328:1332 powerful tools 1319:1332 powerful tools for biological studies on MUC1 O-glycan structures 1319:1383 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 3 61 with antibodies 458:467 arg1 specificities 512:524 designed carbohydrate specificities 490:524 designed carbohydrate specificities 490:524 In this study, using an MUC1 glycopeptide library, we established two novel anti-MUC1 monoclonal antibodies (1B2 and 12D10) with designed carbohydrate specificities. 30023556 2 62 theme low 267:269 arg1 specificities 271:283 low specificities 267:283 low specificities toward O-glycan structures and/or low affinity toward a monovalent epitope 267:358 However, most of them show low specificities toward O-glycan structures and/or low affinity toward a monovalent epitope. 30023556 1 63 theme anti-MUC1 149:157 arg1 antibodies 160:169 Numerous anti-mucin 1 (anti-MUC1) antibodies 126:169 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures 126:209 Numerous anti-mucin 1 (anti-MUC1) antibodies that recognize O-glycan core structures have already been developed. 30023556 8 64 theme 20-mer 1226:1231 arg1 glycopeptide 1233:1244 20-mer glycopeptide 1226:1244 not only native MUC1 but also 20-mer glycopeptide 1196:1244 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 5 65 theme R 786:786 arg1 motif 788:792 the PDT*R motif 778:792 the PDT*R motif (the asterisk represents an O-glycosylation site) 778:842 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 5 66 gly O-glycosylation 822:836 arg2 site 838:841 an O-glycosylation site 819:841 an O-glycosylation site 819:841 Both antibodies recognized specific O-glycan structures at the PDT*R motif (the asterisk represents an O-glycosylation site). 30023556 7 67 gly glycopeptides 1046:1058 arg2 glycopeptides 1046:1058 glycopeptides 1046:1058 glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue 1046:1139 Neither of them bound to glycopeptides with core 2 O-glycans that have GlcNAc at the O-6 position of the GalNAc residue. 30023556 0 68 theme Carbohydrate 66:77 arg1 Specificities 79:91 Designed Carbohydrate Specificities 57:91 Designed Carbohydrate Specificities 57:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 8 69 with binding 1185:1191 arg1 epitope 1264:1270 a monovalent epitope 1251:1270 a monovalent epitope 1251:1270 Furthermore, 1B2 and 12D10 showed a strong binding to not only native MUC1 but also 20-mer glycopeptide with a monovalent epitope. 30023556 0 70 theme Novel 14:18 arg1 Antibodies 41:50 Novel Anti-MUC1 Monoclonal Antibodies 14:50 Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities 14:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 0 71 with Antibodies 41:50 arg1 Specificities 79:91 Designed Carbohydrate Specificities 57:91 Designed Carbohydrate Specificities 57:91 Generation of Novel Anti-MUC1 Monoclonal Antibodies with Designed Carbohydrate Specificities Using MUC1 Glycopeptide Library. 30023556 9 72 theme biological 1338:1347 arg1 studies 1349:1355 biological studies 1338:1355 biological studies on MUC1 O-glycan structures 1338:1383 These anti-MUC1 antibodies should thus become powerful tools for biological studies on MUC1 O-glycan structures. 30023556 6 73 dep residue 919:925 arg1 T 932:932 T 932:932 T 932:932 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 73 dep residue 919:925 arg1 23ST 939:942 23ST 939:942 23ST 939:942 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 30023556 6 73 dep residue 919:925 arg1 Tn 928:929 Tn 928:929 Tn 928:929 1B2 recognized O-glycans with an unsubstituted O-6 position of the GalNAc residue (Tn, T, and 23ST), whereas 12D10 recognized Neu5Ac at the same position (STn, 26ST, and dST). 26869421 2 0 theme polyethylene 468:479 arg1 glycol 481:486 polyethylene glycol 468:486 polyethylene glycol precipitation 468:500 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 0 1 from Impact 16:21 arg1 Profiles 88:95 Physical Stability Profiles 69:95 Physical Stability Profiles of IgG1-Fc Glycoforms 69:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 7 2 gly glycoforms 1323:1332 arg1 IgG1-Fc 1315:1321 the 4 IgG1-Fc glycoforms 1309:1332 the 4 IgG1-Fc glycoforms 1309:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 1 3 from issue 168:172 arg1 series 133:138 a series 131:138 a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 131:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 4 theme biosimilarity 241:253 arg1 analysis 255:262 biosimilarity analysis 241:262 biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 241:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 3 5 theme high-throughput 691:705 arg1 techniques 719:728 multiple high-throughput biophysical techniques 682:728 multiple high-throughput biophysical techniques 682:728 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 3 6 gly glycoproteins 764:776 arg1 glycoproteins 764:776 the Fc glycoproteins 757:776 the Fc glycoproteins 757:776 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 3 7 dep formulations 801:812 arg1 NaCl 815:818 NaCl 815:818 NaCl 815:818 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 3 7 dep formulations 801:812 arg1 formulations 801:812 2 formulations 799:812 2 formulations (NaCl and sucrose) 799:831 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 3 7 dep formulations 801:812 arg1 sucrose 824:830 sucrose 824:830 sucrose 824:830 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 8 8 theme biosimilarity 1451:1463 arg1 analysis 1465:1472 biosimilarity analysis 1451:1472 biosimilarity analysis 1451:1472 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 0 9 theme Glycoforms 108:117 arg1 Profiles 88:95 Physical Stability Profiles 69:95 Physical Stability Profiles of IgG1-Fc Glycoforms 69:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 2 10 theme oligosaccharide 648:662 arg1 structures 664:673 the N297-linked oligosaccharide structures 632:673 the N297-linked oligosaccharide structures 632:673 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 4 11 theme data 914:917 arg1 sets 919:922 The data sets 910:922 The data sets 910:922 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 0 12 theme IgG1-Fc 100:106 arg1 Glycoforms 108:117 IgG1-Fc Glycoforms 100:117 IgG1-Fc Glycoforms 100:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 2 13 theme scanning 567:574 arg1 pH 589:590 pH 4.5 589:594 pH 4.5 589:594 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 13 theme scanning 567:574 arg1 calorimetry 576:586 differential scanning calorimetry 554:586 differential scanning calorimetry (pH 4.5) 554:595 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 1 14 theme high 265:268 arg1 mannose-Fc 270:279 high mannose-Fc 265:279 high mannose-Fc 265:279 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 15 from articles 143:150 arg1 issue 168:172 this special issue 155:172 this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 155:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 7 16 theme oligosaccharide 1363:1377 arg1 structure 1379:1387 oligosaccharide structure 1363:1387 oligosaccharide structure 1363:1387 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 3 17 theme pH 885:886 arg1 conditions 898:907 pH (4.0-7.5) conditions 885:907 pH (4.0-7.5) conditions 885:907 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 1 18 theme special 160:166 arg1 issue 168:172 this special issue 155:172 this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 155:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 4 19 theme phase 965:969 arg1 diagrams 971:978 3-index empirical phase diagrams 947:978 3-index empirical phase diagrams 947:978 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 1 20 theme work 348:351 arg1 comparisons 356:366 comparisons 356:366 comparisons of their physical properties 356:395 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 20 theme work 348:351 arg1 focus 334:338 the focus 330:338 the focus of this work 330:351 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 2 21 link N297-linked 636:646 arg1 structures 664:673 the N297-linked oligosaccharide structures 632:673 the N297-linked oligosaccharide structures 632:673 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 22 theme structures 664:673 arg1 size 624:627 size 624:627 size of the N297-linked oligosaccharide structures 624:673 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 5 23 theme salt 1116:1119 arg1 formulation 1122:1132 the sucrose (vs. salt) formulation 1099:1132 formulation 1122:1132 Each glycoform showed improved stability in the sucrose (vs. salt) formulation. 26869421 4 24 theme radar 984:988 arg1 charts 990:995 radar charts 984:995 radar charts 984:995 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 5 25 theme improved 1077:1084 arg1 stability 1086:1094 improved stability 1077:1094 improved stability 1077:1094 Each glycoform showed improved stability in the sucrose (vs. salt) formulation. 26869421 1 26 from series 133:138 arg1 issue 168:172 this special issue 155:172 this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 155:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 2 27 theme apparent 420:427 arg1 pH 503:504 pH 4.5, 6.0 503:513 pH 503:504 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 27 theme apparent 420:427 arg1 solubility 429:438 apparent solubility 420:438 apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) 420:514 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 27 theme apparent 420:427 arg1 activity 455:462 thermodynamic activity 441:462 thermodynamic activity 441:462 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 8 28 theme companion 1493:1501 arg1 article 1503:1509 a companion article 1491:1509 a companion article 1491:1509 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 1 29 gly glycoforms 208:217 arg1 IgG1-Fc 200:206 4 well-defined IgG1-Fc glycoforms 185:217 4 well-defined IgG1-Fc glycoforms 185:217 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 2 30 theme N297-linked 636:646 arg1 structures 664:673 the N297-linked oligosaccharide structures 632:673 the N297-linked oligosaccharide structures 632:673 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 4 31 used used 929:932 arg2 sets 919:922 The data sets 910:922 The data sets 910:922 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 7 32 with correlation 1346:1356 arg1 structure 1379:1387 oligosaccharide structure 1363:1387 oligosaccharide structure 1363:1387 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 8 33 used used 1410:1413 arg2 sets 1401:1404 These data sets 1390:1404 These data sets 1390:1404 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 2 34 theme lower 520:524 arg1 stability 541:549 lower conformational stability 520:549 lower conformational stability 520:549 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 4 35 theme empirical 955:963 arg1 diagrams 971:978 3-index empirical phase diagrams 947:978 3-index empirical phase diagrams 947:978 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 3 36 theme conditions 898:907 arg1 range 847:851 a wide range 840:851 a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions 840:907 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 1 37 theme well-defined 187:198 arg1 system 230:235 a model system 222:235 a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 222:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 37 theme well-defined 187:198 arg1 glycoforms 208:217 4 well-defined IgG1-Fc glycoforms 185:217 4 well-defined IgG1-Fc glycoforms 185:217 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 3 38 theme physical 735:742 arg1 stability 744:752 the physical stability 731:752 the physical stability of the Fc glycoproteins 731:776 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 2 39 theme thermodynamic 441:453 arg1 solubility 429:438 apparent solubility 420:438 apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) 420:514 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 39 theme thermodynamic 441:453 arg1 activity 455:462 thermodynamic activity 441:462 thermodynamic activity 441:462 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 1 40 theme IgG1-Fc 200:206 arg1 system 230:235 a model system 222:235 a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 222:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 40 theme IgG1-Fc 200:206 arg1 glycoforms 208:217 4 well-defined IgG1-Fc glycoforms 185:217 4 well-defined IgG1-Fc glycoforms 185:217 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 0 41 theme Oligosaccharide 39:53 arg1 Structures 55:64 Well-Defined Oligosaccharide Structures 26:64 Well-Defined Oligosaccharide Structures 26:64 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 8 42 theme data 1396:1399 arg1 sets 1401:1404 These data sets 1390:1404 These data sets 1390:1404 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 7 43 theme glycoforms 1323:1332 arg1 profiles 1297:1304 the overall physical stability profiles 1266:1304 the overall physical stability profiles of the 4 IgG1-Fc glycoforms 1266:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 4 44 theme stability 1044:1052 arg1 regions 1014:1020 the regions 1010:1020 the regions of protein structural stability 1010:1052 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 0 45 theme Well-Defined 26:37 arg1 Structures 55:64 Well-Defined Oligosaccharide Structures 26:64 Well-Defined Oligosaccharide Structures 26:64 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 4 46 theme 3-index 947:953 arg1 diagrams 971:978 3-index empirical phase diagrams 947:978 3-index empirical phase diagrams 947:978 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 7 47 theme overall 1270:1276 arg1 profiles 1297:1304 the overall physical stability profiles 1266:1304 the overall physical stability profiles of the 4 IgG1-Fc glycoforms 1266:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 8 48 dep Kim 1514:1516 arg1 al. 1521:1523 Kim et al. 1514:1523 Kim et al. in this issue 1514:1537 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 3 49 theme glycoproteins 764:776 arg1 stability 744:752 the physical stability 731:752 the physical stability of the Fc glycoproteins 731:776 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 2 50 theme glycol 481:486 arg1 precipitation 488:500 polyethylene glycol precipitation 468:500 polyethylene glycol precipitation 468:500 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 51 theme differential 554:565 arg1 pH 589:590 pH 4.5 589:594 pH 4.5 589:594 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 2 51 theme differential 554:565 arg1 calorimetry 576:586 differential scanning calorimetry 554:586 differential scanning calorimetry (pH 4.5) 554:595 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 1 52 theme physical 377:384 arg1 properties 386:395 their physical properties 371:395 their physical properties 371:395 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 6 53 theme relative 1179:1186 arg1 stability 1188:1196 the highest relative stability 1167:1196 the highest relative stability 1167:1196 The HM-Fc and Man5-Fc displayed the highest relative stability, followed by GlcNAc-Fc, with N297Q-Fc being the least stable. 26869421 3 54 theme biophysical 707:717 arg1 techniques 719:728 multiple high-throughput biophysical techniques 682:728 multiple high-throughput biophysical techniques 682:728 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 0 55 theme Structures 55:64 arg1 Impact 16:21 the Impact 12:21 the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms 12:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 3 56 theme Fc 761:762 arg1 glycoproteins 764:776 the Fc glycoproteins 757:776 the Fc glycoproteins 757:776 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 1 57 theme properties 386:395 arg1 comparisons 356:366 comparisons 356:366 comparisons of their physical properties 356:395 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 57 theme properties 386:395 arg1 focus 334:338 the focus 330:338 the focus of this work 330:351 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 7 58 theme IgG1-Fc 1315:1321 arg1 glycoforms 1323:1332 the 4 IgG1-Fc glycoforms 1309:1332 the 4 IgG1-Fc glycoforms 1309:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 7 59 theme physical 1278:1285 arg1 profiles 1297:1304 the overall physical stability profiles 1266:1304 the overall physical stability profiles of the 4 IgG1-Fc glycoforms 1266:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 4 60 theme structural 1033:1042 arg1 stability 1044:1052 protein structural stability 1025:1052 protein structural stability 1025:1052 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 1 61 theme aglycosylated 314:326 arg1 N297Q-Fc 305:312 N297Q-Fc aglycosylated 305:326 N297Q-Fc aglycosylated 305:326 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 3 62 theme wide 842:845 arg1 range 847:851 a wide range 840:851 a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions 840:907 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 2 63 theme conformational 526:539 arg1 stability 541:549 lower conformational stability 520:549 lower conformational stability 520:549 A trend of decreasing apparent solubility (thermodynamic activity) by polyethylene glycol precipitation (pH 4.5, 6.0) and lower conformational stability by differential scanning calorimetry (pH 4.5) was observed with reducing size of the N297-linked oligosaccharide structures. 26869421 8 64 theme mathematical 1428:1439 arg1 model 1441:1445 a mathematical model 1426:1445 a mathematical model for biosimilarity analysis 1426:1472 These data sets are used to develop a mathematical model for biosimilarity analysis (as described in a companion article by Kim et al. in this issue). 26869421 1 65 theme series 133:138 arg1 part 123:126 part 123:126 part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 123:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 66 theme model 224:228 arg1 system 230:235 a model system 222:235 a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 222:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 66 theme model 224:228 arg1 glycoforms 208:217 4 well-defined IgG1-Fc glycoforms 185:217 4 well-defined IgG1-Fc glycoforms 185:217 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 0 67 theme Stability 78:86 arg1 Profiles 88:95 Physical Stability Profiles 69:95 Physical Stability Profiles of IgG1-Fc Glycoforms 69:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 1 68 dep analysis 255:262 arg1 Man5-Fc 282:288 Man5-Fc 282:288 Man5-Fc 282:288 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 68 dep analysis 255:262 arg1 mannose-Fc 270:279 high mannose-Fc 265:279 high mannose-Fc 265:279 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 68 dep analysis 255:262 arg1 N297Q-Fc 305:312 N297Q-Fc aglycosylated 305:326 N297Q-Fc aglycosylated 305:326 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 68 dep analysis 255:262 arg1 GlcNAc-Fc 291:299 GlcNAc-Fc 291:299 GlcNAc-Fc 291:299 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 7 69 theme stability 1287:1295 arg1 profiles 1297:1304 the overall physical stability profiles 1266:1304 the overall physical stability profiles of the 4 IgG1-Fc glycoforms 1266:1332 Thus, the overall physical stability profiles of the 4 IgG1-Fc glycoforms also show a correlation with oligosaccharide structure. 26869421 6 70 dep relative 1179:1186 arg1 highest 1171:1177 highest 1171:1177 highest 1171:1177 The HM-Fc and Man5-Fc displayed the highest relative stability, followed by GlcNAc-Fc, with N297Q-Fc being the least stable. 26869421 4 71 theme protein 1025:1031 arg1 stability 1044:1052 protein structural stability 1025:1052 protein structural stability 1025:1052 The data sets were used to construct 3-index empirical phase diagrams and radar charts to visualize the regions of protein structural stability. 26869421 0 72 theme Physical 69:76 arg1 Profiles 88:95 Physical Stability Profiles 69:95 Physical Stability Profiles of IgG1-Fc Glycoforms 69:117 Correlating the Impact of Well-Defined Oligosaccharide Structures on Physical Stability Profiles of IgG1-Fc Glycoforms. 26869421 3 73 theme multiple 682:689 arg1 techniques 719:728 multiple high-throughput biophysical techniques 682:728 multiple high-throughput biophysical techniques 682:728 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26869421 1 74 gly aglycosylated 314:326 arg1 N297Q-Fc 305:312 N297Q-Fc aglycosylated 305:326 N297Q-Fc aglycosylated 305:326 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 1 75 theme articles 143:150 arg1 series 133:138 a series 131:138 a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated) 131:327 As part of a series of articles in this special issue describing 4 well-defined IgG1-Fc glycoforms as a model system for biosimilarity analysis (high mannose-Fc, Man5-Fc, GlcNAc-Fc and N297Q-Fc aglycosylated), the focus of this work is comparisons of their physical properties. 26869421 3 76 theme temperatures 856:867 arg1 range 847:851 a wide range 840:851 a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions 840:907 Using multiple high-throughput biophysical techniques, the physical stability of the Fc glycoproteins was then measured in 2 formulations (NaCl and sucrose) across a wide range of temperatures (10°C-90°C) and pH (4.0-7.5) conditions. 26272370 7 0 theme fundamental 1376:1386 arg1 insights 1388:1395 first fundamental insights 1370:1395 first fundamental insights into the TE671 glycosylation potential 1370:1434 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 9 1 theme complex 1681:1687 arg1 structures 1720:1729 complex and highly sialylated N-glycan structures 1681:1729 complex and highly sialylated N-glycan structures 1681:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. 26272370 1 2 theme sectors 286:292 arg1 market 127:132 The market 123:132 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) 123:229 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 2 theme sectors 286:292 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 2 theme sectors 286:292 arg1 one 234:236 one 234:236 one 234:236 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 7 3 theme desorption/ionization 1294:1314 arg1 spectrometry 1348:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 4 4 theme TE671 718:722 arg1 line 729:732 the human TE671 cell line 708:732 the human TE671 cell line 708:732 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 4 5 gly glycoproteins 693:705 arg1 glycoproteins 693:705 recombinant glycoproteins 681:705 recombinant glycoproteins 681:705 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 3 6 theme product 605:611 arg1 immunogenicity 613:626 product immunogenicity 605:626 product immunogenicity 605:626 Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. 26272370 6 7 theme serum-free 1036:1045 arg1 expression 1047:1056 serum-free expression 1036:1056 serum-free expression 1036:1056 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 8 theme procedure 1022:1030 arg1 optimization 984:995 The optimization 980:995 The optimization of the cell transfection procedure and serum-free expression 980:1056 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 5 9 theme initial 762:768 arg1 analysis 770:777 the initial analysis 758:777 the initial analysis of cell membrane proteins 758:803 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 8 10 dep in 1493:1494 arg1 particular 1496:1505 particular 1496:1505 particular 1496:1505 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 6 11 theme cell 1004:1007 arg1 procedure 1022:1030 the cell transfection procedure 1000:1030 the cell transfection procedure 1000:1030 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 5 12 gly glycoproteins 938:950 arg1 glycoproteins 938:950 two model glycoproteins 928:950 two model glycoproteins of therapeutical relevance 928:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 8 13 theme TE671 1618:1622 arg1 cultivations 1629:1640 the respective TE671 cell cultivations 1603:1640 the respective TE671 cell cultivations 1603:1640 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 8 14 theme protein 1445:1451 arg1 pattern 1462:1468 protein specific pattern 1445:1468 protein specific pattern 1445:1468 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 10 15 theme cell 1781:1784 arg1 line 1786:1789 the TE671 cell line 1771:1789 the TE671 cell line 1771:1789 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 10 15 theme cell 1781:1784 arg1 alternative 1804:1814 a serious alternative 1794:1814 a serious alternative to other existing human expression systems 1794:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 1 16 theme therapeutic 137:147 arg1 glycoproteins 149:161 therapeutic glycoproteins 137:161 therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) 137:229 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 16 theme therapeutic 137:147 arg1 cytokines 207:215 cytokines 207:215 cytokines 207:215 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 16 theme therapeutic 137:147 arg1 antibodies 195:204 antibodies 195:204 antibodies 195:204 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 16 theme therapeutic 137:147 arg1 factors 186:192 coagulation factors 174:192 coagulation factors 174:192 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 16 theme therapeutic 137:147 arg1 hormones 221:228 hormones 221:228 hormones 221:228 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 8 17 theme cultivations 1629:1640 arg1 conditions 1589:1598 the medium conditions 1578:1598 the medium conditions of the respective TE671 cell cultivations 1578:1640 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 8 18 theme strong 1471:1476 arg1 distinctions 1478:1489 strong distinctions 1471:1489 strong distinctions - in particular for N-glycan fucosylation and sialylation - 1471:1549 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 7 19 theme first 1370:1374 arg1 insights 1388:1395 first fundamental insights 1370:1395 first fundamental insights into the TE671 glycosylation potential 1370:1434 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 0 20 theme rhabdomyosarcoma 10:25 arg1 line 32:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 5 21 theme glycoproteins 938:950 arg1 expression 914:923 the recombinant expression 898:923 the recombinant expression of two model glycoproteins of therapeutical relevance 898:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 6 22 theme serine 1082:1087 arg1 inhibitor 1098:1106 the human serine protease inhibitor alpha-1-antitrypsin 1072:1126 the human serine protease inhibitor alpha-1-antitrypsin (A1AT) 1072:1133 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 22 theme serine 1082:1087 arg1 A1AT 1129:1132 A1AT 1129:1132 A1AT 1129:1132 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 5 23 theme promising 814:822 arg1 glycosylation 824:836 a promising glycosylation 812:836 a promising glycosylation of TE671 cells 812:851 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 5 24 theme therapeutical 955:967 arg1 relevance 969:977 therapeutical relevance 955:977 therapeutical relevance 955:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 9 25 theme N-glycan 1711:1718 arg1 structures 1720:1729 complex and highly sialylated N-glycan structures 1681:1729 complex and highly sialylated N-glycan structures 1681:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. 26272370 8 26 theme N-glycan 1511:1518 arg1 fucosylation 1520:1531 N-glycan fucosylation 1511:1531 N-glycan fucosylation 1511:1531 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 10 27 theme other 1819:1823 arg1 systems 1851:1857 other existing human expression systems 1819:1857 other existing human expression systems 1819:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 5 28 theme membrane 787:794 arg1 proteins 796:803 cell membrane proteins 782:803 cell membrane proteins 782:803 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 0 29 gly glycosylated 90:101 arg1 biopharmaceuticals 103:120 glycosylated biopharmaceuticals 90:120 glycosylated biopharmaceuticals 90:120 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 6 30 theme cytokine 1157:1164 arg1 GM-CSF 1216:1221 GM-CSF 1216:1221 GM-CSF 1216:1221 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 30 theme cytokine 1157:1164 arg1 factor 1208:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor 1139:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) 1139:1222 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 5 31 theme biotechnological 857:872 arg1 purposes 874:881 biotechnological purposes 857:881 biotechnological purposes 857:881 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 2 32 theme particular 473:482 arg1 glycosylation 484:496 particular glycosylation 473:496 particular glycosylation 473:496 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 9 33 gly sialylated 1700:1709 arg1 structures 1720:1729 complex and highly sialylated N-glycan structures 1681:1729 complex and highly sialylated N-glycan structures 1681:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. 26272370 2 34 dep expensive 370:378 arg1 complex 396:402 complex 396:402 complex 396:402 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 7 35 theme matrix-assisted 1272:1286 arg1 spectrometry 1348:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 1 36 theme biopharmaceutical 301:317 arg1 industry 319:326 the biopharmaceutical industry 297:326 the biopharmaceutical industry 297:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 10 37 theme human 1834:1838 arg1 systems 1851:1857 other existing human expression systems 1819:1857 other existing human expression systems 1819:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 2 38 theme post-translational 436:453 arg1 modifications 455:467 desired post-translational modifications 428:467 desired post-translational modifications 428:467 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 7 39 theme proteins 1260:1267 arg1 analyses 1234:1241 N-glycan analyses 1225:1241 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1225:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 8 40 dep distinctions 1478:1489 arg1 in 1493:1494 in 1493:1494 in 1493:1494 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 0 41 theme production 66:75 arg1 platform 77:84 an innovative production platform 52:84 an innovative production platform for glycosylated biopharmaceuticals 52:120 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 5 42 theme cells 847:851 arg1 glycosylation 824:836 a promising glycosylation 812:836 a promising glycosylation of TE671 cells 812:851 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 8 43 theme medium 1582:1587 arg1 conditions 1589:1598 the medium conditions 1578:1598 the medium conditions of the respective TE671 cell cultivations 1578:1640 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 7 44 theme glycosylation 1412:1424 arg1 potential 1426:1434 the TE671 glycosylation potential 1402:1434 the TE671 glycosylation potential 1402:1434 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 10 45 theme existing 1825:1832 arg1 systems 1851:1857 other existing human expression systems 1819:1857 other existing human expression systems 1819:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 1 46 theme profitable 245:254 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 0 47 theme innovative 55:64 arg1 platform 77:84 an innovative production platform 52:84 an innovative production platform for glycosylated biopharmaceuticals 52:120 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 4 48 theme novel 651:655 arg1 platform 668:675 a novel production platform 649:675 a novel production platform for recombinant glycoproteins 649:705 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 7 49 theme mass 1343:1346 arg1 spectrometry 1348:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 7 50 theme N-glycan 1225:1232 arg1 analyses 1234:1241 N-glycan analyses 1225:1241 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1225:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 3 51 theme ligand 539:544 arg1 binding 546:552 ligand binding 539:552 ligand binding 539:552 Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. 26272370 1 52 theme fast-growing 257:268 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 6 53 theme colony-stimulating 1189:1206 arg1 GM-CSF 1216:1221 GM-CSF 1216:1221 GM-CSF 1216:1221 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 53 theme colony-stimulating 1189:1206 arg1 factor 1208:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor 1139:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) 1139:1222 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 1 54 theme challenging 274:284 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 5 55 theme recombinant 902:912 arg1 expression 914:923 the recombinant expression 898:923 the recombinant expression of two model glycoproteins of therapeutical relevance 898:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 10 56 theme TE671 1775:1779 arg1 line 1786:1789 the TE671 cell line 1771:1789 the TE671 cell line 1771:1789 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 10 56 theme TE671 1775:1779 arg1 alternative 1804:1814 a serious alternative 1794:1814 a serious alternative to other existing human expression systems 1794:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 2 57 theme cell 348:351 arg1 culture 353:359 mammalian cell culture 338:359 mammalian cell culture 338:359 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 6 58 theme expression 1047:1056 arg1 optimization 984:995 The optimization 980:995 The optimization of the cell transfection procedure and serum-free expression 980:1056 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 4 59 theme human 712:716 arg1 line 729:732 the human TE671 cell line 708:732 the human TE671 cell line 708:732 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 3 60 theme biological 582:591 arg1 activity 593:600 biological activity 582:600 biological activity 582:600 Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. 26272370 2 61 theme mammalian 338:346 arg1 culture 353:359 mammalian cell culture 338:359 mammalian cell culture 338:359 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 8 62 theme respective 1607:1616 arg1 cultivations 1629:1640 the respective TE671 cell cultivations 1603:1640 the respective TE671 cell cultivations 1603:1640 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 6 63 theme transfection 1009:1020 arg1 procedure 1022:1030 the cell transfection procedure 1000:1030 the cell transfection procedure 1000:1030 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 8 64 theme cell 1624:1627 arg1 cultivations 1629:1640 the respective TE671 cell cultivations 1603:1640 the respective TE671 cell cultivations 1603:1640 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 4 65 theme recombinant 681:691 arg1 glycoproteins 693:705 recombinant glycoproteins 681:705 recombinant glycoproteins 681:705 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 5 66 theme relevance 969:977 arg1 glycoproteins 938:950 two model glycoproteins 928:950 two model glycoproteins of therapeutical relevance 928:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 1 67 theme glycoproteins 149:161 arg1 one 234:236 one 234:236 one 234:236 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 67 theme glycoproteins 149:161 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 67 theme glycoproteins 149:161 arg1 market 127:132 The market 123:132 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) 123:229 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 6 68 theme alpha-1-antitrypsin 1108:1126 arg1 inhibitor 1098:1106 the human serine protease inhibitor alpha-1-antitrypsin 1072:1126 the human serine protease inhibitor alpha-1-antitrypsin (A1AT) 1072:1133 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 68 theme alpha-1-antitrypsin 1108:1126 arg1 A1AT 1129:1132 A1AT 1129:1132 A1AT 1129:1132 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 8 69 theme specific 1453:1460 arg1 pattern 1462:1468 protein specific pattern 1445:1468 protein specific pattern 1445:1468 Besides protein specific pattern, strong distinctions - in particular for N-glycan fucosylation and sialylation - were observed depending on the medium conditions of the respective TE671 cell cultivations. 26272370 5 70 theme proteins 796:803 arg1 analysis 770:777 the initial analysis 758:777 the initial analysis of cell membrane proteins 758:803 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 5 71 theme model 932:936 arg1 glycoproteins 938:950 two model glycoproteins 928:950 two model glycoproteins of therapeutical relevance 928:977 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 6 72 theme protease 1089:1096 arg1 inhibitor 1098:1106 the human serine protease inhibitor alpha-1-antitrypsin 1072:1126 the human serine protease inhibitor alpha-1-antitrypsin (A1AT) 1072:1133 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 72 theme protease 1089:1096 arg1 A1AT 1129:1132 A1AT 1129:1132 A1AT 1129:1132 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 4 73 theme cell 724:727 arg1 line 729:732 the human TE671 cell line 708:732 the human TE671 cell line 708:732 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 0 74 theme glycosylated 90:101 arg1 biopharmaceuticals 103:120 glycosylated biopharmaceuticals 90:120 glycosylated biopharmaceuticals 90:120 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 6 75 theme human 1076:1080 arg1 inhibitor 1098:1106 the human serine protease inhibitor alpha-1-antitrypsin 1072:1126 the human serine protease inhibitor alpha-1-antitrypsin (A1AT) 1072:1133 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 75 theme human 1076:1080 arg1 A1AT 1129:1132 A1AT 1129:1132 A1AT 1129:1132 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 7 76 theme MALDI-TOF 1332:1340 arg1 spectrometry 1348:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 9 77 theme sialylated 1700:1709 arg1 structures 1720:1729 complex and highly sialylated N-glycan structures 1681:1729 complex and highly sialylated N-glycan structures 1681:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. 26272370 5 78 theme cell 782:785 arg1 proteins 796:803 cell membrane proteins 782:803 cell membrane proteins 782:803 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 6 79 theme granulocyte-macrophage 1166:1187 arg1 GM-CSF 1216:1221 GM-CSF 1216:1221 GM-CSF 1216:1221 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 79 theme granulocyte-macrophage 1166:1187 arg1 factor 1208:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor 1139:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) 1139:1222 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 2 80 from issue 510:514 arg1 glycosylation 484:496 particular glycosylation 473:496 particular glycosylation 473:496 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 0 81 theme human 4:8 arg1 line 32:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 6 82 theme hematopoietic 1143:1155 arg1 GM-CSF 1216:1221 GM-CSF 1216:1221 GM-CSF 1216:1221 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 6 82 theme hematopoietic 1143:1155 arg1 factor 1208:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor 1139:1213 the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) 1139:1222 The optimization of the cell transfection procedure and serum-free expression succeeded for the human serine protease inhibitor alpha-1-antitrypsin (A1AT) and the hematopoietic cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF). 26272370 10 83 theme serious 1796:1802 arg1 line 1786:1789 the TE671 cell line 1771:1789 the TE671 cell line 1771:1789 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 10 83 theme serious 1796:1802 arg1 alternative 1804:1814 a serious alternative 1794:1814 a serious alternative to other existing human expression systems 1794:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 0 84 theme cell 27:30 arg1 line 32:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line 0:35 The human rhabdomyosarcoma cell line TE671--Towards an innovative production platform for glycosylated biopharmaceuticals. 26272370 10 85 theme expression 1840:1849 arg1 systems 1851:1857 other existing human expression systems 1819:1857 other existing human expression systems 1819:1857 Our results demonstrate the TE671 cell line as a serious alternative to other existing human expression systems. 26272370 7 86 theme purified 1251:1258 arg1 proteins 1260:1267 both purified proteins 1246:1267 both purified proteins 1246:1267 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 1 87 theme industry 319:326 arg1 sectors 286:292 the profitable, fast-growing and challenging sectors 241:292 the profitable, fast-growing and challenging sectors of the biopharmaceutical industry 241:326 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 2 88 theme major 504:508 arg1 ability 409:415 the ability to produce desired post-translational modifications 405:467 the ability to produce desired post-translational modifications 405:467 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 2 88 theme major 504:508 arg1 issue 510:514 a major issue 502:514 a major issue 502:514 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 2 89 theme desired 428:434 arg1 modifications 455:467 desired post-translational modifications 428:467 desired post-translational modifications 428:467 Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-translational modifications, in particular glycosylation, is a major issue. 26272370 5 90 theme TE671 841:845 arg1 cells 847:851 TE671 cells 841:851 TE671 cells 841:851 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 1 91 gly glycoproteins 149:161 arg1 glycoproteins 149:161 therapeutic glycoproteins 137:161 therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) 137:229 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 91 gly glycoproteins 149:161 arg1 cytokines 207:215 cytokines 207:215 cytokines 207:215 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 91 gly glycoproteins 149:161 arg1 antibodies 195:204 antibodies 195:204 antibodies 195:204 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 91 gly glycoproteins 149:161 arg1 factors 186:192 coagulation factors 174:192 coagulation factors 174:192 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 91 gly glycoproteins 149:161 arg1 hormones 221:228 hormones 221:228 hormones 221:228 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 1 92 theme coagulation 174:184 arg1 factors 186:192 coagulation factors 174:192 coagulation factors 174:192 The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. 26272370 7 93 theme TE671 1406:1410 arg1 potential 1426:1434 the TE671 glycosylation potential 1402:1434 the TE671 glycosylation potential 1402:1434 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26272370 5 94 gly glycosylation 824:836 arg1 cells 847:851 TE671 cells 841:851 TE671 cells 841:851 Since the initial analysis of cell membrane proteins showed a promising glycosylation of TE671 cells for biotechnological purposes, we focused on the recombinant expression of two model glycoproteins of therapeutical relevance. 26272370 3 95 theme serum 555:559 arg1 half-life 561:569 serum half-life 555:569 serum half-life 555:569 Glycans can influence ligand binding, serum half-life as well as biological activity or product immunogenicity. 26272370 9 96 theme cell 1647:1650 arg1 ability 1659:1665 The cell line's ability to synthesize complex and highly sialylated N-glycan structures 1643:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures 1643:1729 The cell line's ability to synthesize complex and highly sialylated N-glycan structures has been shown. 26272370 4 97 theme production 657:666 arg1 platform 668:675 a novel production platform 649:675 a novel production platform for recombinant glycoproteins 649:705 Aiming to establish a novel production platform for recombinant glycoproteins, the human TE671 cell line was investigated. 26272370 7 98 theme laser 1288:1292 arg1 spectrometry 1348:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry 1272:1359 N-glycan analyses of both purified proteins by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided first fundamental insights into the TE671 glycosylation potential. 26250826 2 0 theme stereoselective 370:384 arg1 reduction 386:394 stereoselective reduction 370:394 stereoselective reduction 370:394 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 1 theme Stille 296:301 arg1 coupling 303:310 a Stille coupling 294:310 a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside 294:434 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 2 theme derhodinosylurdamycin 615:635 arg1 A 637:637 derhodinosylurdamycin A 615:637 derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies 615:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 1 3 theme first 69:73 arg1 synthesis 81:89 The first total synthesis 65:89 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, 65:154 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, has been described. 26250826 1 4 theme total 75:79 arg1 synthesis 81:89 The first total synthesis 65:89 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, 65:154 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, has been described. 26250826 2 5 theme A 529:529 arg1 preparation 492:502 the preparation 488:502 the preparation of derhodinosylurdamycin A 488:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 6 theme chemical 582:589 arg1 synthesis 591:599 the chemical synthesis 578:599 the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies 578:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 2 7 theme 2-deoxy 410:416 arg1 β-C-arylglycoside 418:434 the 2-deoxy β-C-arylglycoside 406:434 the 2-deoxy β-C-arylglycoside 406:434 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 8 theme glycal 315:320 arg1 stannane 322:329 glycal stannane 315:329 glycal stannane 315:329 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 9 theme stannane 322:329 arg1 aglycon 285:291 the tetracyclic angular aglycon 261:291 the tetracyclic angular aglycon 261:291 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 9 theme stannane 322:329 arg1 glycosylation 470:482 a late-stage stereoselective glycosylation 441:482 a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A 441:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 9 theme stannane 322:329 arg1 coupling 303:310 a Stille coupling 294:310 a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside 294:434 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 0 10 theme Total 0:4 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A 0:62 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A. 26250826 1 11 theme derhodinosylurdamycin 94:114 arg1 antibiotic 144:153 antibiotic 144:153 antibiotic 144:153 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, has been described. 26250826 1 11 theme derhodinosylurdamycin 94:114 arg1 A 116:116 derhodinosylurdamycin A 94:116 derhodinosylurdamycin A 94:116 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, has been described. 26250826 0 12 theme Antitumor 19:27 arg1 A 62:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A. 26250826 1 13 theme A 116:116 arg1 synthesis 81:89 The first total synthesis 65:89 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, 65:154 The first total synthesis of derhodinosylurdamycin A, an angucycline antitumor antibiotic, has been described. 26250826 2 14 theme late-stage 443:452 arg1 glycosylation 470:482 a late-stage stereoselective glycosylation 441:482 a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A 441:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 15 theme stereoselective 454:468 arg1 glycosylation 470:482 a late-stage stereoselective glycosylation 441:482 a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A 441:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 0 16 theme Derhodinosylurdamycin 40:60 arg1 A 62:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A. 26250826 3 17 theme 2-deoxy 655:661 arg1 subunits 669:676 diverse 2-deoxy sugar subunits 647:676 diverse 2-deoxy sugar subunits 647:676 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 0 18 theme Antibiotic 29:38 arg1 A 62:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Antitumor Antibiotic Derhodinosylurdamycin A 19:62 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A. 26250826 2 19 theme derhodinosylurdamycin 507:527 arg1 A 529:529 derhodinosylurdamycin A 507:529 derhodinosylurdamycin A 507:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 20 theme activity 696:703 arg1 studies 718:724 activity relationship studies 696:724 activity relationship studies 696:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 3 21 theme sugar 663:667 arg1 subunits 669:676 diverse 2-deoxy sugar subunits 647:676 diverse 2-deoxy sugar subunits 647:676 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 3 22 theme relationship 705:716 arg1 studies 718:724 activity relationship studies 696:724 activity relationship studies 696:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 2 23 theme pinacol 231:237 arg1 coupling 239:246 pinacol coupling 231:246 pinacol coupling 231:246 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 24 theme synthetic 537:545 arg1 strategy 547:554 This synthetic strategy 532:554 This synthetic strategy 532:554 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 0 25 theme A 62:62 arg1 Synthesis 6:14 Total Synthesis 0:14 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A 0:62 Total Synthesis of Antitumor Antibiotic Derhodinosylurdamycin A. 26250826 3 26 theme diverse 647:653 arg1 subunits 669:676 diverse 2-deoxy sugar subunits 647:676 diverse 2-deoxy sugar subunits 647:676 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 2 27 theme aryliodide 347:356 arg1 aglycon 285:291 the tetracyclic angular aglycon 261:291 the tetracyclic angular aglycon 261:291 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 27 theme aryliodide 347:356 arg1 glycosylation 470:482 a late-stage stereoselective glycosylation 441:482 a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A 441:529 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 27 theme aryliodide 347:356 arg1 coupling 303:310 a Stille coupling 294:310 a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside 294:434 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 28 theme angular 277:283 arg1 aglycon 285:291 the tetracyclic angular aglycon 261:291 the tetracyclic angular aglycon 261:291 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 29 theme tetracyclic 335:345 arg1 aryliodide 347:356 tetracyclic aryliodide 335:356 tetracyclic aryliodide 335:356 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 30 theme analogs 604:610 arg1 synthesis 591:599 the chemical synthesis 578:599 the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies 578:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 26250826 2 31 theme Hauser 201:206 arg1 annulation 208:217 a Hauser annulation 199:217 a Hauser annulation followed by pinacol coupling 199:246 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 32 theme tetracyclic 265:275 arg1 aglycon 285:291 the tetracyclic angular aglycon 261:291 the tetracyclic angular aglycon 261:291 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 33 gly glycosylation 470:482 arg1 aryliodide 347:356 tetracyclic aryliodide 335:356 tetracyclic aryliodide 335:356 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 2 33 gly glycosylation 470:482 arg1 stannane 322:329 glycal stannane 315:329 glycal stannane 315:329 The synthesis features a Hauser annulation followed by pinacol coupling to construct the tetracyclic angular aglycon, a Stille coupling of glycal stannane and tetracyclic aryliodide followed by stereoselective reduction to afford the 2-deoxy β-C-arylglycoside, and a late-stage stereoselective glycosylation for the preparation of derhodinosylurdamycin A. 26250826 3 34 theme A 637:637 arg1 analogs 604:610 analogs 604:610 analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies 604:724 This synthetic strategy should be amenable to the chemical synthesis of analogs of derhodinosylurdamycin A bearing diverse 2-deoxy sugar subunits for structure and activity relationship studies. 25117244 6 0 theme surface 1101:1107 arg1 glycomes 1109:1116 intact bacterial cell surface glycomes 1079:1116 intact bacterial cell surface glycomes 1079:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 3 1 from acids 525:529 arg1 acids 604:608 lipoteichoic acids 591:608 lipoteichoic acids in their phospholipid bilayer 591:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 3 1 from acids 525:529 arg1 glycans 560:566 the peptide glycans 548:566 the peptide glycans of their cell wall 548:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 2 2 from glycolipids 340:350 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 9 3 theme binding 1446:1452 arg1 affinity 1454:1461 its lectin binding affinity 1435:1461 its lectin binding affinity 1435:1461 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 11 4 theme binding 1581:1587 arg1 unique 1629:1634 unique 1629:1634 unique 1629:1634 The lectin binding profile of most strains was found to be unique. 25117244 11 4 theme binding 1581:1587 arg1 profile 1589:1595 The lectin binding profile 1570:1595 The lectin binding profile of most strains 1570:1611 The lectin binding profile of most strains was found to be unique. 25117244 3 5 theme wall 582:585 arg1 acids 604:608 lipoteichoic acids 591:608 lipoteichoic acids in their phospholipid bilayer 591:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 3 5 theme wall 582:585 arg1 glycans 560:566 the peptide glycans 548:566 the peptide glycans of their cell wall 548:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 1 6 theme glycan 200:205 arg1 analysis 207:214 comprehensive glycan analysis 186:214 comprehensive glycan analysis 186:214 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 2 7 theme evanescent-field 226:241 arg1 technique 266:274 evanescent-field fluorescence detection technique 226:274 evanescent-field fluorescence detection technique 226:274 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 7 8 with incubation 1189:1198 arg1 microarray 1216:1225 the lectin microarray 1205:1225 the lectin microarray 1205:1225 The method involves labeling bacterial cells with SYTOX Orange before incubation with the lectin microarray. 25117244 4 9 theme glycan 650:655 arg1 structures 657:666 Specific glycan structures 641:666 Specific glycan structures 641:666 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 8 10 theme fluorescence 1303:1314 arg1 scanner 1316:1322 an evanescent-field fluorescence scanner 1283:1322 an evanescent-field fluorescence scanner in a liquid phase 1283:1340 After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. 25117244 2 11 theme detection 256:264 arg1 technique 266:274 evanescent-field fluorescence detection technique 226:274 evanescent-field fluorescence detection technique 226:274 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 8 12 theme bound 1243:1247 arg1 cells 1249:1253 bound cells 1243:1253 bound cells 1243:1253 After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. 25117244 3 13 theme lipoteichoic 591:602 arg1 acids 604:608 lipoteichoic acids 591:608 lipoteichoic acids in their phospholipid bilayer 591:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 5 14 theme bacteria 929:936 arg1 structure 916:924 glycosylation structure 902:924 glycosylation structure of bacteria 902:936 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 4 15 theme lactic 755:760 arg1 bacteria 767:774 lactic acid bacteria 755:774 lactic acid bacteria 755:774 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 8 16 theme liquid 1329:1334 arg1 phase 1336:1340 a liquid phase 1327:1340 a liquid phase 1327:1340 After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. 25117244 4 17 theme bacteria 713:720 arg1 characteristic 672:685 characteristic 672:685 characteristic 672:685 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 0 18 theme casei/paracasei 69:83 arg1 strains 85:91 Lactobacillus casei/paracasei strains 55:91 Lactobacillus casei/paracasei strains 55:91 Application of lectin microarray to bacteria including Lactobacillus casei/paracasei strains. 25117244 3 19 theme phospholipid 619:630 arg1 bilayer 632:638 their phospholipid bilayer 613:638 their phospholipid bilayer 613:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 1 20 theme simple 152:157 arg1 technique 172:180 a simple and powerful technique 150:180 a simple and powerful technique for comprehensive glycan analysis 150:214 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 6 21 theme method 1011:1016 arg1 development 973:983 the development 969:983 the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes 969:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 9 22 from bacteria 1395:1402 arg1 h 1372:1372 3 h 1370:1372 3 h from putting labeled bacteria 1370:1402 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 1 23 theme powerful 163:170 arg1 technique 172:180 a simple and powerful technique 150:180 a simple and powerful technique for comprehensive glycan analysis 150:214 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 12 24 theme bacteria 1700:1707 arg1 profiling 1687:1695 rapid profiling 1681:1695 rapid profiling of bacteria 1681:1707 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 3 25 from glycans 560:566 arg1 bilayer 632:638 their phospholipid bilayer 613:638 their phospholipid bilayer 613:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 12 26 theme novel 1662:1666 arg1 strategy 1668:1675 a novel strategy 1660:1675 a novel strategy for rapid profiling of bacteria 1660:1707 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 7 27 theme lectin 1209:1214 arg1 microarray 1216:1225 the lectin microarray 1205:1225 the lectin microarray 1205:1225 The method involves labeling bacterial cells with SYTOX Orange before incubation with the lectin microarray. 25117244 6 28 theme intact 1079:1084 arg1 glycomes 1109:1116 intact bacterial cell surface glycomes 1079:1116 intact bacterial cell surface glycomes 1079:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 4 29 theme bacteria 767:774 arg1 moiety 745:750 polysaccharides moiety 729:750 polysaccharides moiety of lactic acid bacteria 729:774 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 2 30 theme cells 445:449 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 0 31 theme lectin 15:20 arg1 microarray 22:31 lectin microarray 15:31 lectin microarray 15:31 Application of lectin microarray to bacteria including Lactobacillus casei/paracasei strains. 25117244 1 32 theme microarray 113:122 arg1 technology 124:133 lectin microarray technology 106:133 lectin microarray technology 106:133 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 12 33 with strains 1760:1766 arg1 relevance 1773:1781 relevance 1773:1781 relevance 1773:1781 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 10 34 theme cell 1499:1502 arg1 glycomes 1512:1519 the cell surface glycomes 1495:1519 the cell surface glycomes from 16 different strains of L. casei/paracasei 1495:1567 Using this method, we compared the cell surface glycomes from 16 different strains of L. casei/paracasei. 25117244 4 35 theme host 831:834 arg1 response 843:850 the host immune response 827:850 the host immune response 827:850 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 2 36 theme live 429:432 arg1 cells 445:449 live eukaryotic cells 429:449 live eukaryotic cells 429:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 7 37 theme SYTOX 1169:1173 arg1 Orange 1175:1180 SYTOX Orange 1169:1180 SYTOX Orange before incubation with the lectin microarray 1169:1225 The method involves labeling bacterial cells with SYTOX Orange before incubation with the lectin microarray. 25117244 6 38 theme microarray 1034:1043 arg1 technology 1045:1054 lectin microarray technology 1027:1054 lectin microarray technology for direct analysis of intact bacterial cell surface glycomes 1027:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 2 39 theme eukaryotic 364:373 arg1 cells 375:379 eukaryotic cells 364:379 eukaryotic cells 364:379 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 9 40 theme labeled 1387:1393 arg1 bacteria 1395:1402 putting labeled bacteria 1379:1402 putting labeled bacteria 1379:1402 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 2 41 from surface 418:424 arg1 analysis 301:308 analysis 301:308 analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells 301:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 11 42 theme most 1600:1603 arg1 strains 1605:1611 most strains 1600:1611 most strains 1600:1611 The lectin binding profile of most strains was found to be unique. 25117244 2 43 gly glycoproteins 322:334 arg1 glycoproteins 322:334 glycoproteins 322:334 glycoproteins 322:334 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 2 44 theme glycoconjugates 390:404 arg1 analysis 301:308 analysis 301:308 analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells 301:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 0 45 theme Lactobacillus 55:67 arg1 strains 85:91 Lactobacillus casei/paracasei strains 55:91 Lactobacillus casei/paracasei strains 55:91 Application of lectin microarray to bacteria including Lactobacillus casei/paracasei strains. 25117244 12 46 theme surface 1814:1820 arg1 glycosylation 1822:1834 surface glycosylation 1814:1834 surface glycosylation 1814:1834 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 6 47 theme glycomes 1109:1116 arg1 analysis 1067:1074 direct analysis 1060:1074 direct analysis of intact bacterial cell surface glycomes 1060:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 6 48 theme cell 1096:1099 arg1 glycomes 1109:1116 intact bacterial cell surface glycomes 1079:1116 intact bacterial cell surface glycomes 1079:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 3 49 theme peptide 552:558 arg1 glycans 560:566 the peptide glycans 548:566 the peptide glycans of their cell wall 548:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 9 50 theme lectin 1439:1444 arg1 affinity 1454:1461 its lectin binding affinity 1435:1461 its lectin binding affinity 1435:1461 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 3 51 from proteins 536:543 arg1 acids 604:608 lipoteichoic acids 591:608 lipoteichoic acids in their phospholipid bilayer 591:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 3 51 from proteins 536:543 arg1 glycans 560:566 the peptide glycans 548:566 the peptide glycans of their cell wall 548:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 1 52 theme comprehensive 186:198 arg1 analysis 207:214 comprehensive glycan analysis 186:214 comprehensive glycan analysis 186:214 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 12 53 theme numerous 1741:1748 arg1 strains 1760:1766 numerous bacterial strains 1741:1766 numerous bacterial strains with relevance to the biological functions of surface glycosylation 1741:1834 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 3 54 from polysaccharides 499:513 arg1 acids 604:608 lipoteichoic acids 591:608 lipoteichoic acids in their phospholipid bilayer 591:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 3 54 from polysaccharides 499:513 arg1 glycans 560:566 the peptide glycans 548:566 the peptide glycans of their cell wall 548:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 3 55 theme cell 577:580 arg1 wall 582:585 their cell wall 571:585 their cell wall 571:585 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 5 56 theme analysis 876:883 arg1 method 866:871 the method 862:871 the method of analysis and knowledge of glycosylation structure of bacteria 862:936 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 5 56 theme analysis 876:883 arg1 limited 942:948 limited 942:948 limited 942:948 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 2 57 from glycoconjugates 390:404 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 4 58 theme many 690:693 arg1 bacteria 713:720 many highly pathogenic bacteria 690:720 many highly pathogenic bacteria 690:720 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 11 59 theme lectin 1574:1579 arg1 unique 1629:1634 unique 1629:1634 unique 1629:1634 The lectin binding profile of most strains was found to be unique. 25117244 11 59 theme lectin 1574:1579 arg1 profile 1589:1595 The lectin binding profile 1570:1595 The lectin binding profile of most strains 1570:1611 The lectin binding profile of most strains was found to be unique. 25117244 2 60 theme glycoproteins 322:334 arg1 analysis 301:308 analysis 301:308 analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells 301:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 5 61 theme structure 916:924 arg1 knowledge 889:897 knowledge 889:897 knowledge 889:897 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 5 61 theme structure 916:924 arg1 analysis 876:883 analysis 876:883 analysis 876:883 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 4 62 theme Specific 641:648 arg1 structures 657:666 Specific glycan structures 641:666 Specific glycan structures 641:666 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 8 63 theme evanescent-field 1286:1301 arg1 scanner 1316:1322 an evanescent-field fluorescence scanner 1283:1322 an evanescent-field fluorescence scanner in a liquid phase 1283:1340 After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. 25117244 2 64 theme fluorescence 243:254 arg1 technique 266:274 evanescent-field fluorescence detection technique 226:274 evanescent-field fluorescence detection technique 226:274 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 4 65 theme polysaccharides 729:743 arg1 moiety 745:750 polysaccharides moiety 729:750 polysaccharides moiety of lactic acid bacteria 729:774 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 12 66 theme bacterial 1750:1758 arg1 strains 1760:1766 numerous bacterial strains 1741:1766 numerous bacterial strains with relevance to the biological functions of surface glycosylation 1741:1834 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 6 67 theme lectin 1027:1032 arg1 technology 1045:1054 lectin microarray technology 1027:1054 lectin microarray technology for direct analysis of intact bacterial cell surface glycomes 1027:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 5 68 theme knowledge 889:897 arg1 method 866:871 the method 862:871 the method of analysis and knowledge of glycosylation structure of bacteria 862:936 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 5 68 theme knowledge 889:897 arg1 limited 942:948 limited 942:948 limited 942:948 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 2 69 from analysis 301:308 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 10 70 from strains 1539:1545 arg1 glycomes 1512:1519 the cell surface glycomes 1495:1519 the cell surface glycomes from 16 different strains of L. casei/paracasei 1495:1567 Using this method, we compared the cell surface glycomes from 16 different strains of L. casei/paracasei. 25117244 4 71 theme pathogenic 702:711 arg1 bacteria 713:720 many highly pathogenic bacteria 690:720 many highly pathogenic bacteria 690:720 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 5 72 theme glycosylation 902:914 arg1 structure 916:924 glycosylation structure 902:924 glycosylation structure of bacteria 902:936 However, the method of analysis and knowledge of glycosylation structure of bacteria are limited. 25117244 12 73 theme rapid 1681:1685 arg1 profiling 1687:1695 rapid profiling 1681:1695 rapid profiling of bacteria 1681:1707 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 6 74 theme sensitive 1001:1009 arg1 method 1011:1016 a simple and sensitive method 988:1016 a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes 988:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 2 75 from glycoproteins 322:334 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 6 76 theme simple 990:995 arg1 method 1011:1016 a simple and sensitive method 988:1016 a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes 988:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 8 77 from scanner 1316:1322 arg1 phase 1336:1340 a liquid phase 1327:1340 a liquid phase 1327:1340 After washing, bound cells are directly detected using an evanescent-field fluorescence scanner in a liquid phase. 25117244 6 78 theme bacterial 1086:1094 arg1 glycomes 1109:1116 intact bacterial cell surface glycomes 1079:1116 intact bacterial cell surface glycomes 1079:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 2 79 theme cell 413:416 arg1 surface 418:424 the cell surface 409:424 the cell surface of live eukaryotic cells 409:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 9 80 theme entire 1347:1352 arg1 procedure 1354:1362 The entire procedure 1343:1362 The entire procedure 1343:1362 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 4 81 theme acid 762:765 arg1 bacteria 767:774 lactic acid bacteria 755:774 lactic acid bacteria 755:774 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 3 82 theme Bacterial 452:460 arg1 cells 462:466 Bacterial cells 452:466 Bacterial cells 452:466 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 0 83 theme microarray 22:31 arg1 Application 0:10 Application 0:10 Application of lectin microarray to bacteria including Lactobacillus casei/paracasei strains. 0:92 Application of lectin microarray to bacteria including Lactobacillus casei/paracasei strains. 25117244 1 84 theme lectin 106:111 arg1 technology 124:133 lectin microarray technology 106:133 lectin microarray technology 106:133 Since 2005, lectin microarray technology has emerged as a simple and powerful technique for comprehensive glycan analysis. 25117244 6 85 theme direct 1060:1065 arg1 analysis 1067:1074 direct analysis 1060:1074 direct analysis of intact bacterial cell surface glycomes 1060:1116 Here, we describe the development of a simple and sensitive method based on lectin microarray technology for direct analysis of intact bacterial cell surface glycomes. 25117244 10 86 theme surface 1504:1510 arg1 glycomes 1512:1519 the cell surface glycomes 1495:1519 the cell surface glycomes from 16 different strains of L. casei/paracasei 1495:1567 Using this method, we compared the cell surface glycomes from 16 different strains of L. casei/paracasei. 25117244 3 87 from acids 604:608 arg1 bilayer 632:638 their phospholipid bilayer 613:638 their phospholipid bilayer 613:638 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 2 88 theme eukaryotic 434:443 arg1 cells 445:449 live eukaryotic cells 429:449 live eukaryotic cells 429:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 11 89 theme strains 1605:1611 arg1 unique 1629:1634 unique 1629:1634 unique 1629:1634 The lectin binding profile of most strains was found to be unique. 25117244 11 89 theme strains 1605:1611 arg1 profile 1589:1595 The lectin binding profile 1570:1595 The lectin binding profile of most strains 1570:1611 The lectin binding profile of most strains was found to be unique. 25117244 7 90 theme bacterial 1148:1156 arg1 cells 1158:1162 bacterial cells 1148:1162 bacterial cells 1148:1162 The method involves labeling bacterial cells with SYTOX Orange before incubation with the lectin microarray. 25117244 9 91 theme putting 1379:1385 arg1 bacteria 1395:1402 putting labeled bacteria 1379:1402 putting labeled bacteria 1379:1402 The entire procedure takes 3 h from putting labeled bacteria on the microarray to profiling its lectin binding affinity. 25117244 12 92 theme glycosylation 1822:1834 arg1 functions 1801:1809 the biological functions 1786:1809 the biological functions of surface glycosylation 1786:1834 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 10 93 theme different 1529:1537 arg1 strains 1539:1545 16 different strains 1526:1545 16 different strains of L. casei/paracasei 1526:1567 Using this method, we compared the cell surface glycomes from 16 different strains of L. casei/paracasei. 25117244 4 94 theme immune 836:841 arg1 response 843:850 the host immune response 827:850 the host immune response 827:850 Specific glycan structures are characteristic of many highly pathogenic bacteria, while polysaccharides moiety of lactic acid bacteria are known to play a role as probiotics to modulate the host immune response. 25117244 3 95 theme teichoic 516:523 arg1 acids 525:529 teichoic acids 516:529 teichoic acids 516:529 Bacterial cells are known to be decorated with polysaccharides, teichoic acids, and proteins in the peptide glycans of their cell wall and lipoteichoic acids in their phospholipid bilayer. 25117244 2 96 theme glycolipids 340:350 arg1 analysis 301:308 analysis 301:308 analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells 301:449 By using evanescent-field fluorescence detection technique, it has been applied for analysis of not only glycoproteins and glycolipids secreted by eukaryotic cells but also glycoconjugates on the cell surface of live eukaryotic cells. 25117244 12 97 theme biological 1790:1799 arg1 functions 1801:1809 the biological functions 1786:1809 the biological functions of surface glycosylation 1786:1834 Our technique provides a novel strategy for rapid profiling of bacteria and enables us to differentiate numerous bacterial strains with relevance to the biological functions of surface glycosylation. 25117244 10 98 theme casei/paracasei 1553:1567 arg1 strains 1539:1545 16 different strains 1526:1545 16 different strains of L. casei/paracasei 1526:1567 Using this method, we compared the cell surface glycomes from 16 different strains of L. casei/paracasei. 24981920 3 0 with glycopeptides 657:669 arg1 chains 687:692 two glycan chains 676:692 two glycan chains towards catalytic hydrogenation conditions 676:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 2 1 contain containing 347:356 arg1 glycopeptide 334:345 a glycosaminoglycan family glycopeptide 307:345 a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 307:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 1 contain containing 347:356 arg2 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 1 contain containing 347:356 arg2 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 1 contain containing 347:356 arg2 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 4 2 theme glycan 825:830 arg1 chain 832:836 the partially deprotected single glycan chain 792:836 the partially deprotected single glycan chain containing glycopeptides 792:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 2 3 theme sulfate 380:386 arg1 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 3 theme sulfate 380:386 arg1 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 3 theme sulfate 380:386 arg1 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 5 4 theme important 1036:1044 arg1 class 1046:1050 this important class 1031:1050 this important class of molecules 1031:1063 This work lays the foundation for preparing other members of this important class of molecules. 24981920 4 5 theme ester-type 940:949 arg1 groups 962:967 the ester-type protecting groups 936:967 the ester-type protecting groups 936:967 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 4 6 gly glycopeptides 849:861 arg2 glycopeptides 849:861 glycopeptides 849:861 glycopeptides 849:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 2 7 theme heparan 372:378 arg1 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 7 theme heparan 372:378 arg1 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 7 theme heparan 372:378 arg1 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 4 8 theme deprotected 806:816 arg1 chain 832:836 the partially deprotected single glycan chain 792:836 the partially deprotected single glycan chain containing glycopeptides 792:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 3 9 dep size 458:461 arg1 the 448:450 the 448:450 the 448:450 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 5 10 theme class 1046:1050 arg1 members 1020:1026 other members 1014:1026 other members of this important class of molecules 1014:1063 This work lays the foundation for preparing other members of this important class of molecules. 24981920 4 11 theme fragments 910:918 arg1 union 882:886 union 882:886 union of the glycan-bearing fragments 882:918 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 4 11 theme fragments 910:918 arg1 cleavage 924:931 cleavage 924:931 cleavage of the ester-type protecting groups 924:967 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 2 12 theme family 327:332 arg1 glycopeptide 334:345 a glycosaminoglycan family glycopeptide 307:345 a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 307:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 4 13 theme groups 962:967 arg1 union 882:886 union 882:886 union of the glycan-bearing fragments 882:918 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 4 13 theme groups 962:967 arg1 cleavage 924:931 cleavage 924:931 cleavage of the ester-type protecting groups 924:967 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 2 14 theme first 288:292 arg1 synthesis 294:302 the first synthesis 284:302 the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 284:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 15 theme glycopeptide 334:345 arg1 synthesis 294:302 the first synthesis 284:302 the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 284:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 3 16 theme molecules 509:517 arg1 size 458:461 large size 452:461 large size 452:461 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 16 theme molecules 509:517 arg1 complexity 489:498 tremendous structural complexity 467:498 tremendous structural complexity 467:498 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 5 17 theme molecules 1055:1063 arg1 class 1046:1050 this important class 1031:1050 this important class of molecules 1031:1063 This work lays the foundation for preparing other members of this important class of molecules. 24981920 4 18 theme protecting 951:960 arg1 groups 962:967 the ester-type protecting groups 936:967 the ester-type protecting groups 936:967 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 1 19 gly glycopeptides 203:215 arg2 glycopeptides 203:215 glycopeptides 203:215 glycopeptides 203:215 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 2 20 theme extracellular 407:419 arg1 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 20 theme extracellular 407:419 arg1 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 20 theme extracellular 407:419 arg1 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 1 21 dep methodologies 161:173 arg1 synthesize 178:187 synthesize 178:187 to synthesize this class of glycopeptides with homogeneous glycans 175:240 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 0 22 theme Chemical 0:7 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of syndecan-3 glycopeptides 0:45 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 2 23 theme glycosaminoglycan 309:325 arg1 glycopeptide 334:345 a glycosaminoglycan family glycopeptide 307:345 a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 307:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 3 24 theme large 452:456 arg1 size 458:461 large size 452:461 large size 452:461 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 0 25 theme syndecan-3 22:31 arg1 glycopeptides 33:45 syndecan-3 glycopeptides 22:45 syndecan-3 glycopeptides 22:45 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 5 26 theme other 1014:1018 arg1 members 1020:1026 other members 1014:1026 other members of this important class of molecules 1014:1063 This work lays the foundation for preparing other members of this important class of molecules. 24981920 1 27 theme ubiquitous 102:111 arg1 presence 113:120 the ubiquitous presence 98:120 the ubiquitous presence of proteoglycans in mammalian systems 98:158 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 1 28 theme glycopeptides 203:215 arg1 class 194:198 this class 189:198 this class of glycopeptides with homogeneous glycans 189:240 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 3 29 theme high 599:602 arg1 sensitivity 604:614 high sensitivity 599:614 high sensitivity to base treatment 599:632 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 30 theme unexpected 529:538 arg1 obstacles 540:548 multiple unexpected obstacles 520:548 multiple unexpected obstacles 520:548 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 31 theme multiple 520:527 arg1 obstacles 540:548 multiple unexpected obstacles 520:548 multiple unexpected obstacles 520:548 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 2 32 theme syndecan-3 431:440 arg1 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 32 theme syndecan-3 431:440 arg1 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 32 theme syndecan-3 431:440 arg1 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 1 33 theme homogeneous 222:232 arg1 glycans 234:240 homogeneous glycans 222:240 homogeneous glycans 222:240 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 0 34 theme glycopeptides 33:45 arg1 synthesis 9:17 Chemical synthesis 0:17 Chemical synthesis of syndecan-3 glycopeptides 0:45 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 3 35 theme structural 478:487 arg1 complexity 489:498 tremendous structural complexity 467:498 tremendous structural complexity 467:498 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 0 36 theme heparan 59:65 arg1 chains 82:87 two heparan sulfate glycan chains 55:87 two heparan sulfate glycan chains 55:87 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 1 37 with class 194:198 arg1 glycans 234:240 homogeneous glycans 222:240 homogeneous glycans 222:240 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 3 38 theme glycan 680:685 arg1 chains 687:692 two glycan chains 676:692 two glycan chains towards catalytic hydrogenation conditions 676:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 39 theme base 619:622 arg1 treatment 624:632 base treatment 619:632 base treatment 619:632 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 2 40 theme different 362:370 arg1 syndecan-3 431:440 syndecan-3 431:440 syndecan-3 431:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 40 theme different 362:370 arg1 domain 421:426 the extracellular domain 403:426 the extracellular domain of syndecan-3 403:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 2 40 theme different 362:370 arg1 chains 388:393 two different heparan sulfate chains 358:393 two different heparan sulfate chains 358:393 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 1 41 attach presence 113:120 arg1 systems 152:158 mammalian systems 142:158 mammalian systems 142:158 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 1 41 attach presence 113:120 arg2 proteoglycans 125:137 proteoglycans 125:137 proteoglycans 125:137 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 4 42 theme glycan-bearing 895:908 arg1 fragments 910:918 the glycan-bearing fragments 891:918 the glycan-bearing fragments 891:918 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 1 43 theme proteoglycans 125:137 arg1 presence 113:120 the ubiquitous presence 98:120 the ubiquitous presence of proteoglycans in mammalian systems 98:158 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 4 44 contain containing 838:847 arg2 glycopeptides 849:861 glycopeptides 849:861 glycopeptides 849:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 4 44 contain containing 838:847 arg1 chain 832:836 the partially deprotected single glycan chain 792:836 the partially deprotected single glycan chain containing glycopeptides 792:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 2 45 gly glycopeptide 334:345 arg2 glycopeptide 334:345 a glycosaminoglycan family glycopeptide 307:345 a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3 307:440 Herein, we report the first synthesis of a glycosaminoglycan family glycopeptide containing two different heparan sulfate chains, namely the extracellular domain of syndecan-3. 24981920 3 46 theme glycopeptides 657:669 arg1 sensitivity 604:614 high sensitivity 599:614 high sensitivity to base treatment 599:632 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 46 theme glycopeptides 657:669 arg1 instability 642:652 the instability 638:652 the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions 638:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 0 47 gly glycopeptides 33:45 arg2 glycopeptides 33:45 syndecan-3 glycopeptides 22:45 syndecan-3 glycopeptides 22:45 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 1 48 from presence 113:120 arg1 systems 152:158 mammalian systems 142:158 mammalian systems 142:158 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 3 49 theme catalytic 702:710 arg1 conditions 726:735 catalytic hydrogenation conditions 702:735 catalytic hydrogenation conditions 702:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 4 50 theme single 818:823 arg1 chain 832:836 the partially deprotected single glycan chain 792:836 the partially deprotected single glycan chain containing glycopeptides 792:861 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 1 51 theme mammalian 142:150 arg1 systems 152:158 mammalian systems 142:158 mammalian systems 142:158 Despite the ubiquitous presence of proteoglycans in mammalian systems, methodologies to synthesize this class of glycopeptides with homogeneous glycans are not well developed. 24981920 0 52 theme glycan 75:80 arg1 chains 82:87 two heparan sulfate glycan chains 55:87 two heparan sulfate glycan chains 55:87 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 24981920 3 53 theme hydrogenation 712:724 arg1 conditions 726:735 catalytic hydrogenation conditions 702:735 catalytic hydrogenation conditions 702:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 3 54 theme tremendous 467:476 arg1 complexity 489:498 tremendous structural complexity 467:498 tremendous structural complexity 467:498 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 4 55 theme successful 740:749 arg1 strategy 751:758 A successful strategy 738:758 A successful strategy 738:758 A successful strategy was established by constructing the partially deprotected single glycan chain containing glycopeptides first, followed by union of the glycan-bearing fragments and cleavage of the ester-type protecting groups. 24981920 3 56 gly glycopeptides 657:669 arg2 glycopeptides 657:669 glycopeptides 657:669 glycopeptides with two glycan chains towards catalytic hydrogenation conditions 657:735 With the large size and tremendous structural complexity of these molecules, multiple unexpected obstacles were encountered during the synthesis, including high sensitivity to base treatment and the instability of glycopeptides with two glycan chains towards catalytic hydrogenation conditions. 24981920 0 57 theme sulfate 67:73 arg1 chains 82:87 two heparan sulfate glycan chains 55:87 two heparan sulfate glycan chains 55:87 Chemical synthesis of syndecan-3 glycopeptides bearing two heparan sulfate glycan chains. 27389966 10 0 theme infection 1476:1484 arg1 glycans 1453:1459 left remodeled host glycans 1433:1459 left remodeled host glycans 1433:1459 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 0 theme infection 1476:1484 arg1 GHs 1404:1406 two keystone GHs 1391:1406 two keystone GHs for internalization 1391:1426 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 0 theme infection 1476:1484 arg1 result 1466:1471 a result 1464:1471 a result of infection 1464:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 6 1 theme N-acetylneuraminic 634:651 arg1 Neu5Ac 659:664 Neu5Ac 659:664 Neu5Ac 659:664 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 1 theme N-acetylneuraminic 634:651 arg1 monosaccharides 617:631 the terminal monosaccharides 604:631 the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) 604:698 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 1 theme N-acetylneuraminic 634:651 arg1 acid 653:656 N-acetylneuraminic acid 634:656 N-acetylneuraminic acid (Neu5Ac) 634:665 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 0 2 theme Glycan 73:78 arg1 Remodeling 80:89 Glycan Remodeling 73:89 Glycan Remodeling 73:89 Salmonella Degrades the Host Glycocalyx Leading to Altered Infection and Glycan Remodeling. 27389966 8 3 theme glycans 1030:1036 arg1 modification 1009:1020 modification 1009:1020 modification of host glycans 1009:1036 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 8 4 theme biosynthesis 976:987 arg1 pathways 989:996 N-glycan biosynthesis pathways 967:996 N-glycan biosynthesis pathways leading to modification of host glycans 967:1036 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 7 5 theme different 857:865 arg1 structures 874:883 different glycan structures 857:883 different glycan structures 857:883 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 4 6 contain contains 363:370 arg1 Salmonella 352:361 Salmonella 352:361 Salmonella 352:361 Salmonella contains 47 glycosyl hydrolases (GHs) that may degrade the glycan. 27389966 4 6 contain contains 363:370 arg2 GHs 396:398 GHs 396:398 GHs 396:398 Salmonella contains 47 glycosyl hydrolases (GHs) that may degrade the glycan. 27389966 4 6 contain contains 363:370 arg2 hydrolases 384:393 47 glycosyl hydrolases 372:393 47 glycosyl hydrolases (GHs) that may degrade the glycan 372:427 Salmonella contains 47 glycosyl hydrolases (GHs) that may degrade the glycan. 27389966 9 7 theme remodeled 1226:1234 arg1 glycans 1236:1242 remodeled glycans 1226:1242 remodeled glycans 1226:1242 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 8 8 theme N-glycan 967:974 arg1 pathways 989:996 N-glycan biosynthesis pathways 967:996 N-glycan biosynthesis pathways leading to modification of host glycans 967:1036 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 7 9 theme glycan 867:872 arg1 structures 874:883 different glycan structures 857:883 different glycan structures 857:883 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 10 10 dep established 1292:1302 arg1 determined 1355:1364 determined 1355:1364 determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection 1355:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 5 11 theme entire 475:480 arg1 complement 485:494 the entire GH complement 471:494 the entire GH complement of Salmonella 471:508 We hypothesized that keystone genes from the entire GH complement of Salmonella are required to degrade glycans to change infection. 27389966 8 12 theme mannose 1069:1075 arg1 content 1077:1083 mannose content 1069:1083 mannose content 1069:1083 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 8 13 theme pathways 989:996 arg1 induction 954:962 the induction 950:962 the induction of N-glycan biosynthesis pathways leading to modification of host glycans 950:1036 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 11 14 theme virulence 1543:1551 arg1 GHs 1522:1524 microbial GHs 1512:1524 microbial GHs 1512:1524 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 11 14 theme virulence 1543:1551 arg1 factors 1553:1559 undiscovered virulence factors 1530:1559 undiscovered virulence factors 1530:1559 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 5 15 theme Salmonella 499:508 arg1 complement 485:494 the entire GH complement 471:494 the entire GH complement of Salmonella 471:508 We hypothesized that keystone genes from the entire GH complement of Salmonella are required to degrade glycans to change infection. 27389966 6 16 theme terminal 608:615 arg1 mannose 679:685 mannose 679:685 mannose 679:685 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 16 theme terminal 608:615 arg1 fucose 692:697 fucose 692:697 fucose 692:697 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 16 theme terminal 608:615 arg1 monosaccharides 617:631 the terminal monosaccharides 604:631 the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) 604:698 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 16 theme terminal 608:615 arg1 acid 653:656 N-acetylneuraminic acid 634:656 N-acetylneuraminic acid (Neu5Ac) 634:665 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 16 theme terminal 608:615 arg1 galactose 668:676 galactose 668:676 galactose 668:676 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 4 17 theme glycosyl 375:382 arg1 GHs 396:398 GHs 396:398 GHs 396:398 Salmonella contains 47 glycosyl hydrolases (GHs) that may degrade the glycan. 27389966 4 17 theme glycosyl 375:382 arg1 hydrolases 384:393 47 glycosyl hydrolases 372:393 47 glycosyl hydrolases (GHs) that may degrade the glycan 372:427 Salmonella contains 47 glycosyl hydrolases (GHs) that may degrade the glycan. 27389966 7 18 theme GHs 788:790 arg1 nanH 802:805 GHs sialidase nanH 788:805 GHs sialidase nanH 788:805 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 8 19 theme host 1025:1028 arg1 glycans 1030:1036 host glycans 1025:1036 host glycans 1025:1036 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 1 20 theme Complex 92:98 arg1 glycans 100:106 Complex glycans 92:106 Complex glycans 92:106 Complex glycans cover the gut epithelial surface to protect the cell from the environment. 27389966 7 21 theme amylase 811:817 arg1 malS 819:822 its two GHs sialidase nanH and amylase malS 780:822 its two GHs sialidase nanH and amylase malS for internalization 780:842 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 0 22 theme Host 24:27 arg1 Glycocalyx 29:38 the Host Glycocalyx 20:38 the Host Glycocalyx Leading to Altered Infection and Glycan Remodeling 20:89 Salmonella Degrades the Host Glycocalyx Leading to Altered Infection and Glycan Remodeling. 27389966 7 23 theme sialidase 792:800 arg1 nanH 802:805 GHs sialidase nanH 788:805 GHs sialidase nanH 788:805 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 10 24 theme remodeled 1438:1446 arg1 GHs 1404:1406 two keystone GHs 1391:1406 two keystone GHs for internalization 1391:1426 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 24 theme remodeled 1438:1446 arg1 glycans 1453:1459 left remodeled host glycans 1433:1459 left remodeled host glycans 1433:1459 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 24 theme remodeled 1438:1446 arg1 result 1466:1471 a result 1464:1471 a result of infection 1464:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 25 theme epithelial 1337:1346 arg1 cells 1348:1352 colonic epithelial cells 1329:1352 colonic epithelial cells 1329:1352 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 9 26 theme expression 1121:1130 arg1 analysis 1132:1139 Gene expression analysis 1116:1139 Gene expression analysis 1116:1139 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 10 27 theme left 1433:1436 arg1 GHs 1404:1406 two keystone GHs 1391:1406 two keystone GHs for internalization 1391:1426 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 27 theme left 1433:1436 arg1 glycans 1453:1459 left remodeled host glycans 1433:1459 left remodeled host glycans 1433:1459 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 27 theme left 1433:1436 arg1 result 1466:1471 a result 1464:1471 a result of infection 1464:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 28 theme colonic 1329:1335 arg1 cells 1348:1352 colonic epithelial cells 1329:1352 colonic epithelial cells 1329:1352 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 7 29 theme nanH 802:805 arg1 malS 819:822 its two GHs sialidase nanH and amylase malS 780:822 its two GHs sialidase nanH and amylase malS for internalization 780:842 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 8 30 theme host 890:893 arg1 glycans 895:901 The host glycans 886:901 The host glycans 886:901 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 3 31 theme glycan 266:271 arg1 degradation 273:283 glycan degradation 266:283 glycan degradation 266:283 While glycan degradation is crucial for infection, this process is inadequately understood. 27389966 11 32 theme undiscovered 1530:1541 arg1 GHs 1522:1524 microbial GHs 1512:1524 microbial GHs 1512:1524 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 11 32 theme undiscovered 1530:1541 arg1 factors 1553:1559 undiscovered virulence factors 1530:1559 undiscovered virulence factors 1530:1559 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 5 33 theme GH 482:483 arg1 complement 485:494 the entire GH complement 471:494 the entire GH complement of Salmonella 471:508 We hypothesized that keystone genes from the entire GH complement of Salmonella are required to degrade glycans to change infection. 27389966 9 34 theme Salmonella 1259:1268 arg1 treatment 1270:1278 Salmonella treatment 1259:1278 Salmonella treatment 1259:1278 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 10 35 theme glycan 1308:1313 arg1 structures 1315:1324 the glycan structures 1304:1324 the glycan structures on colonic epithelial cells 1304:1352 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 0 36 theme Altered 51:57 arg1 Infection 59:67 Altered Infection 51:67 Altered Infection 51:67 Salmonella Degrades the Host Glycocalyx Leading to Altered Infection and Glycan Remodeling. 27389966 2 37 theme glycan 218:223 arg1 layer 225:229 the glycan layer 214:229 the glycan layer 214:229 Invasive pathogens must breach the glycan layer before initiating infection. 27389966 6 38 dep monosaccharides 617:631 arg1 fucose 692:697 fucose 692:697 fucose 692:697 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 38 dep monosaccharides 617:631 arg1 monosaccharides 617:631 the terminal monosaccharides 604:631 the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) 604:698 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 38 dep monosaccharides 617:631 arg1 galactose 668:676 galactose 668:676 galactose 668:676 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 38 dep monosaccharides 617:631 arg1 acid 653:656 N-acetylneuraminic acid 634:656 N-acetylneuraminic acid (Neu5Ac) 634:665 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 38 dep monosaccharides 617:631 arg1 mannose 679:685 mannose 679:685 mannose 679:685 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 6 38 dep monosaccharides 617:631 arg1 Neu5Ac 659:664 Neu5Ac 659:664 Neu5Ac 659:664 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 10 39 theme host 1448:1451 arg1 GHs 1404:1406 two keystone GHs 1391:1406 two keystone GHs for internalization 1391:1426 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 39 theme host 1448:1451 arg1 glycans 1453:1459 left remodeled host glycans 1433:1459 left remodeled host glycans 1433:1459 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 39 theme host 1448:1451 arg1 result 1466:1471 a result 1464:1471 a result of infection 1464:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 9 40 theme host 1160:1163 arg1 cell 1165:1168 the host cell 1156:1168 the host cell 1156:1168 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 1 41 theme gut 118:120 arg1 surface 133:139 the gut epithelial surface 114:139 the gut epithelial surface to protect the cell from the environment 114:180 Complex glycans cover the gut epithelial surface to protect the cell from the environment. 27389966 8 42 theme Salmonella 923:932 arg1 association 934:944 Salmonella association 923:944 Salmonella association 923:944 The host glycans were altered during Salmonella association via the induction of N-glycan biosynthesis pathways leading to modification of host glycans by increasing fucosylation and mannose content, while decreasing sialylation. 27389966 1 43 theme epithelial 122:131 arg1 surface 133:139 the gut epithelial surface 114:139 the gut epithelial surface to protect the cell from the environment 114:180 Complex glycans cover the gut epithelial surface to protect the cell from the environment. 27389966 6 44 dep significantly 704:716 arg1 p < 0.05 719:726 p < 0.05 719:726 p < 0.05 719:726 This study determined that GHs recognize the terminal monosaccharides (N-acetylneuraminic acid (Neu5Ac), galactose, mannose, and fucose) and significantly (p < 0.05) alter infection. 27389966 9 45 theme Gene 1116:1119 arg1 analysis 1132:1139 Gene expression analysis 1116:1139 Gene expression analysis 1116:1139 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 7 46 used used 775:778 arg2 Salmonella 764:773 Salmonella 764:773 Salmonella 764:773 During infection, Salmonella used its two GHs sialidase nanH and amylase malS for internalization by targeting different glycan structures. 27389966 5 47 theme keystone 451:458 arg1 genes 460:464 keystone genes 451:464 keystone genes from the entire GH complement of Salmonella 451:508 We hypothesized that keystone genes from the entire GH complement of Salmonella are required to degrade glycans to change infection. 27389966 9 48 dep treatment 1270:1278 arg1 response 1247:1254 response 1247:1254 response 1247:1254 Gene expression analysis indicated that the host cell responded by regulating more than 50 genes resulting in remodeled glycans in response to Salmonella treatment. 27389966 10 49 from structures 1315:1324 arg1 cells 1348:1352 colonic epithelial cells 1329:1352 colonic epithelial cells 1329:1352 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 2 50 theme Invasive 183:190 arg1 pathogens 192:200 Invasive pathogens 183:200 Invasive pathogens 183:200 Invasive pathogens must breach the glycan layer before initiating infection. 27389966 5 51 from complement 485:494 arg1 genes 460:464 keystone genes 451:464 keystone genes from the entire GH complement of Salmonella 451:508 We hypothesized that keystone genes from the entire GH complement of Salmonella are required to degrade glycans to change infection. 27389966 11 52 theme microbial 1512:1520 arg1 GHs 1522:1524 microbial GHs 1512:1524 microbial GHs 1512:1524 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 11 52 theme microbial 1512:1520 arg1 factors 1553:1559 undiscovered virulence factors 1530:1559 undiscovered virulence factors 1530:1559 These data indicate that microbial GHs are undiscovered virulence factors. 27389966 10 53 theme keystone 1395:1402 arg1 glycans 1453:1459 left remodeled host glycans 1433:1459 left remodeled host glycans 1433:1459 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 53 theme keystone 1395:1402 arg1 GHs 1404:1406 two keystone GHs 1391:1406 two keystone GHs for internalization 1391:1426 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 27389966 10 53 theme keystone 1395:1402 arg1 result 1466:1471 a result 1464:1471 a result of infection 1464:1484 This study established the glycan structures on colonic epithelial cells, determined that Salmonella required two keystone GHs for internalization, and left remodeled host glycans as a result of infection. 29275050 9 0 theme crowded 1436:1442 arg1 structures 1444:1453 the sterically crowded structures 1421:1453 the sterically crowded structures 1421:1453 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 8 1 theme su-LeX 1173:1178 arg1 oxazoline 1180:1188 the excess su-LeX oxazoline 1162:1188 the excess su-LeX oxazoline 1162:1188 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 2 2 theme % 356:356 arg1 cosolvent 366:374 10% organic cosolvent 354:374 10% organic cosolvent 354:374 The addition of 10% organic cosolvent reduced the activity for the enzymatic transglycosylation. 29275050 0 3 theme sugar 74:78 arg1 oxazolines 80:89 the sugar oxazolines 70:89 the sugar oxazolines having keratan sulfate repeating units 70:128 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 0 4 from specificity 9:19 arg1 transglycosylation 45:62 the transglycosylation 41:62 the transglycosylation 41:62 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 7 5 theme steric 1052:1057 arg1 hindrance 1059:1067 the steric hindrance 1048:1067 the steric hindrance 1048:1067 These results strongly suggest that the steric hindrance exists around the (+3)(+4) subsites in KSase II. 29275050 2 6 theme 10 354:355 arg1 % 356:356 % 356:356 % 356:356 The addition of 10% organic cosolvent reduced the activity for the enzymatic transglycosylation. 29275050 0 7 theme keratan 98:104 arg1 sulfate 106:112 keratan sulfate 98:112 keratan sulfate repeating units 98:128 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 6 8 theme corresponding 977:989 arg1 dimers 991:996 the corresponding dimers 973:996 the corresponding dimers respectively 973:1009 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 1 9 contain having 298:303 arg1 derivatives 286:296 the oxazoline derivatives 272:296 the oxazoline derivatives having keratan sulfate repeating units 272:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 1 9 contain having 298:303 arg2 units 331:335 keratan sulfate repeating units 305:335 keratan sulfate repeating units 305:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 9 10 theme +3 1495:1496 arg1 tunnel-like 1542:1552 tunnel-like 1542:1552 tunnel-like 1542:1552 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 9 10 theme +3 1495:1496 arg1 subsites 1503:1510 the (+3)(+4) subsites 1490:1510 the (+3)(+4) subsites 1490:1510 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 5 11 theme su-LeX 783:788 arg1 acceptor 791:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 8 12 theme catalytic 1333:1341 arg1 center 1343:1348 the catalytic center 1329:1348 the catalytic center hampering processive shift of this pentasaccharide 1329:1399 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 7 13 from subsites 1096:1103 arg1 KSase 1108:1112 KSase II 1108:1115 KSase II 1108:1115 These results strongly suggest that the steric hindrance exists around the (+3)(+4) subsites in KSase II. 29275050 6 14 from addition 853:860 arg1 oligomerized 957:968 oligomerized 957:968 oligomerized 957:968 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 6 14 from addition 853:860 arg1 derivatives 882:892 both the oxazoline derivatives 863:892 both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc 863:933 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 8 15 theme sole 1258:1261 arg1 product 1282:1288 the sole transglycosylation product 1254:1288 the sole transglycosylation product 1254:1288 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 15 theme sole 1258:1261 arg1 pentasaccharide 1235:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 5 16 gly transglycosylation 699:716 arg1 donor 745:749 the su-LacNAc oxazoline donor 721:749 the su-LacNAc oxazoline donor 721:749 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 0 17 theme Reaction 0:7 arg1 specificity 9:19 Reaction specificity 0:19 Reaction specificity of keratanase II in the transglycosylation 0:62 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 6 18 from oligomerized 957:968 arg1 addition 853:860 addition 853:860 addition 853:860 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 5 19 with transglycosylation 699:716 arg1 acceptor 791:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 5 20 theme X 780:780 arg1 acceptor 791:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 3 21 theme oxazoline 439:447 arg1 derivative 449:458 The oxazoline derivative 435:458 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) 435:507 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) was processively oligomerized to the corresponding hexamer or longer by the enzyme. 29275050 3 21 theme oxazoline 439:447 arg1 oligomerized 526:537 oligomerized 526:537 oligomerized 526:537 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) was processively oligomerized to the corresponding hexamer or longer by the enzyme. 29275050 0 22 theme keratanase 24:33 arg1 specificity 9:19 Reaction specificity 0:19 Reaction specificity of keratanase II in the transglycosylation 0:62 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 1 23 theme oxazoline 276:284 arg1 derivatives 286:296 the oxazoline derivatives 272:296 the oxazoline derivatives having keratan sulfate repeating units 272:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 5 24 theme donor 745:749 arg1 transglycosylation 699:716 the transglycosylation 695:716 the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor 695:798 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 8 25 theme transglycosylation 1263:1280 arg1 product 1282:1288 the sole transglycosylation product 1254:1288 the sole transglycosylation product 1254:1288 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 25 theme transglycosylation 1263:1280 arg1 pentasaccharide 1235:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 7 26 theme +3 1088:1089 arg1 subsites 1096:1103 the (+3)(+4) subsites 1083:1103 the (+3)(+4) subsites in KSase II 1083:1115 These results strongly suggest that the steric hindrance exists around the (+3)(+4) subsites in KSase II. 29275050 1 27 theme transglycosylation 163:180 arg1 specificity 144:154 The reaction specificity 131:154 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) 131:249 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 4 28 theme numbered 663:670 arg1 subsites 672:679 the large positively numbered subsites 642:679 the large positively numbered subsites 642:679 This result strongly implies that the enzyme has the large positively numbered subsites. 29275050 5 29 theme su-LacNAc-su-LeX 816:831 arg1 pentasaccharide 833:847 the su-LacNAc-su-LeX pentasaccharide 812:847 the su-LacNAc-su-LeX pentasaccharide 812:847 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 2 30 theme enzymatic 405:413 arg1 transglycosylation 415:432 the enzymatic transglycosylation 401:432 the enzymatic transglycosylation 401:432 The addition of 10% organic cosolvent reduced the activity for the enzymatic transglycosylation. 29275050 8 31 theme KSase 1131:1135 arg1 reaction 1150:1157 KSase II-catalyzed reaction 1131:1157 KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc 1131:1207 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 3 32 theme 6-O-sulfonato-N-acetyllactosamine 463:495 arg1 derivative 449:458 The oxazoline derivative 435:458 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) 435:507 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) was processively oligomerized to the corresponding hexamer or longer by the enzyme. 29275050 3 32 theme 6-O-sulfonato-N-acetyllactosamine 463:495 arg1 oligomerized 526:537 oligomerized 526:537 oligomerized 526:537 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) was processively oligomerized to the corresponding hexamer or longer by the enzyme. 29275050 9 33 theme catalytic 1462:1470 arg1 center 1472:1477 the catalytic center 1458:1477 the catalytic center 1458:1477 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 1 34 theme keratan 305:311 arg1 sulfate 313:319 keratan sulfate 305:319 keratan sulfate repeating units 305:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 5 35 theme 6-O-sulfonato-Lewis 760:778 arg1 acceptor 791:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 the 6-O-sulfonato-Lewis X (su-LeX) acceptor 756:798 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 6 36 theme 6,6'-di-O-sulfonato-LacNAc 908:933 arg1 derivatives 882:892 both the oxazoline derivatives 863:892 both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc 863:933 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 6 36 theme 6,6'-di-O-sulfonato-LacNAc 908:933 arg1 oligomerized 957:968 oligomerized 957:968 oligomerized 957:968 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 1 37 from circulans 223:231 arg1 keratanase 195:204 keratanase II 195:207 keratanase II from Bacillus circulans KsT202 (KSase II) 195:249 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 1 38 theme sulfate 313:319 arg1 units 331:335 keratan sulfate repeating units 305:335 keratan sulfate repeating units 305:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 8 39 theme excess 1166:1171 arg1 oxazoline 1180:1188 the excess su-LeX oxazoline 1162:1188 the excess su-LeX oxazoline 1162:1188 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 40 theme oxazoline 1180:1188 arg1 reaction 1150:1157 KSase II-catalyzed reaction 1131:1157 KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc 1131:1207 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 1 41 theme repeating 321:329 arg1 units 331:335 keratan sulfate repeating units 305:335 keratan sulfate repeating units 305:335 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 8 42 theme II-catalyzed 1137:1148 arg1 reaction 1150:1157 KSase II-catalyzed reaction 1131:1157 KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc 1131:1207 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 6 43 theme su-LeX 897:902 arg1 derivatives 882:892 both the oxazoline derivatives 863:892 both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc 863:933 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 6 43 theme su-LeX 897:902 arg1 oligomerized 957:968 oligomerized 957:968 oligomerized 957:968 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 0 44 contain having 91:96 arg1 oxazolines 80:89 the sugar oxazolines 70:89 the sugar oxazolines having keratan sulfate repeating units 70:128 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 0 44 contain having 91:96 arg2 sulfate 106:112 keratan sulfate 98:112 keratan sulfate repeating units 98:128 Reaction specificity of keratanase II in the transglycosylation using the sugar oxazolines having keratan sulfate repeating units. 29275050 5 45 theme su-LacNAc 725:733 arg1 donor 745:749 the su-LacNAc oxazoline donor 721:749 the su-LacNAc oxazoline donor 721:749 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 3 46 theme corresponding 546:558 arg1 hexamer 560:566 the corresponding hexamer 542:566 the corresponding hexamer 542:566 The oxazoline derivative of 6-O-sulfonato-N-acetyllactosamine (su-LacNAc) was processively oligomerized to the corresponding hexamer or longer by the enzyme. 29275050 8 47 theme pentasaccharide 1385:1399 arg1 shift 1371:1375 processive shift 1360:1375 processive shift of this pentasaccharide 1360:1399 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 48 with reaction 1150:1157 arg1 su-LacNAc 1199:1207 the su-LacNAc 1195:1207 the su-LacNAc 1195:1207 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 49 theme su-LeX-su-LacNAc 1218:1233 arg1 product 1282:1288 the sole transglycosylation product 1254:1288 the sole transglycosylation product 1254:1288 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 49 theme su-LeX-su-LacNAc 1218:1233 arg1 pentasaccharide 1235:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 the su-LeX-su-LacNAc pentasaccharide 1214:1249 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 5 50 theme oxazoline 735:743 arg1 donor 745:749 the su-LacNAc oxazoline donor 721:749 the su-LacNAc oxazoline donor 721:749 In contrast, the transglycosylation of the su-LacNAc oxazoline donor with the 6-O-sulfonato-Lewis X (su-LeX) acceptor solely gave the su-LacNAc-su-LeX pentasaccharide. 29275050 1 51 theme reaction 135:142 arg1 specificity 144:154 The reaction specificity 131:154 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) 131:249 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 1 52 theme KSase 241:245 arg1 II 247:248 KSase II 241:248 KSase II 241:248 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 1 52 theme KSase 241:245 arg1 circulans 223:231 Bacillus circulans 214:231 Bacillus circulans KsT202 (KSase II) 214:249 The reaction specificity of the transglycosylation catalyzed by keratanase II from Bacillus circulans KsT202 (KSase II) was studied by using the oxazoline derivatives having keratan sulfate repeating units. 29275050 6 53 theme oxazoline 872:880 arg1 derivatives 882:892 both the oxazoline derivatives 863:892 both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc 863:933 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 6 53 theme oxazoline 872:880 arg1 oligomerized 957:968 oligomerized 957:968 oligomerized 957:968 In addition, both the oxazoline derivatives of su-LeX and 6,6'-di-O-sulfonato-LacNAc have been exclusively oligomerized to the corresponding dimers respectively. 29275050 4 54 contain has 638:640 arg1 enzyme 631:636 the enzyme 627:636 the enzyme 627:636 This result strongly implies that the enzyme has the large positively numbered subsites. 29275050 4 54 contain has 638:640 arg2 subsites 672:679 the large positively numbered subsites 642:679 the large positively numbered subsites 642:679 This result strongly implies that the enzyme has the large positively numbered subsites. 29275050 9 55 contain has 1417:1419 arg2 structures 1444:1453 the sterically crowded structures 1421:1453 the sterically crowded structures 1421:1453 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 9 55 contain has 1417:1419 arg1 KSase 1408:1412 KSase II 1408:1415 KSase II 1408:1415 Thus, KSase II has the sterically crowded structures at the catalytic center and around the (+3)(+4) subsites, which are all expected to be tunnel-like. 29275050 2 56 theme cosolvent 366:374 arg1 addition 342:349 The addition 338:349 The addition of 10% organic cosolvent 338:374 The addition of 10% organic cosolvent reduced the activity for the enzymatic transglycosylation. 29275050 8 57 theme steric 1309:1314 arg1 hindrance 1316:1324 the steric hindrance 1305:1324 the steric hindrance 1305:1324 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 8 58 theme processive 1360:1369 arg1 shift 1371:1375 processive shift 1360:1375 processive shift of this pentasaccharide 1360:1399 Furthermore, KSase II-catalyzed reaction of the excess su-LeX oxazoline with the su-LacNAc gave the su-LeX-su-LacNAc pentasaccharide as the sole transglycosylation product, also implying the steric hindrance at the catalytic center hampering processive shift of this pentasaccharide. 29275050 2 59 theme organic 358:364 arg1 cosolvent 366:374 10% organic cosolvent 354:374 10% organic cosolvent 354:374 The addition of 10% organic cosolvent reduced the activity for the enzymatic transglycosylation. 29275050 4 60 theme large 646:650 arg1 subsites 672:679 the large positively numbered subsites 642:679 the large positively numbered subsites 642:679 This result strongly implies that the enzyme has the large positively numbered subsites. 28966766 5 0 theme acids 952:956 arg1 structures 918:927 the structures 914:927 the structures of O-glycans and amino acids local to the glycosylation site 914:988 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 0 1 theme carbohydrate-binding 73:92 arg1 module 94:99 a carbohydrate-binding module 71:99 a carbohydrate-binding module 71:99 Molecular-scale features that govern the effects of O-glycosylation on a carbohydrate-binding module. 28966766 4 2 theme pronounced 727:736 arg1 effects 738:744 the most pronounced effects 718:744 the most pronounced effects of O-glycosylation in this system 718:778 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 3 3 theme model 385:389 arg1 glycoprotein 391:402 a small model glycoprotein 377:402 a small model glycoprotein 377:402 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 3 4 theme glycan 420:425 arg1 structure 427:435 the glycan structure 416:435 the glycan structure 416:435 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 2 5 from importance 209:218 arg1 molecular 223:231 molecular 223:231 molecular 223:231 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 2 5 from importance 209:218 arg1 biology 246:252 cellular biology 237:252 cellular biology 237:252 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 2 6 theme biomolecular 311:322 arg1 structure 324:332 biomolecular structure 311:332 biomolecular structure 311:332 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 5 7 theme glycosylation 971:983 arg1 site 985:988 the glycosylation site 967:988 the glycosylation site 967:988 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 1 8 theme life 191:194 arg1 kingdoms 179:186 all kingdoms 175:186 all kingdoms of life 175:194 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 3 9 theme small 379:383 arg1 glycoprotein 391:402 a small model glycoprotein 377:402 a small model glycoprotein 377:402 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 0 10 from effects 41:47 arg1 module 94:99 a carbohydrate-binding module 71:99 a carbohydrate-binding module 71:99 Molecular-scale features that govern the effects of O-glycosylation on a carbohydrate-binding module. 28966766 3 11 theme physical 565:572 arg1 stability 574:582 physical stability 565:582 physical stability 565:582 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 4 12 theme studied 686:692 arg1 features 694:701 all the studied features 678:701 all the studied features 678:701 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 5 13 theme proteins 845:852 arg1 possibility 820:830 the possibility 816:830 the possibility of designing proteins with multiple improved properties 816:886 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 5 14 theme local 958:962 arg1 structures 918:927 the structures 914:927 the structures of O-glycans and amino acids local to the glycosylation site 914:988 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 1 15 from modification 159:170 arg1 kingdoms 179:186 all kingdoms 175:186 all kingdoms of life 175:194 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 3 16 theme amino 447:451 arg1 residues 458:465 amino acid residues 447:465 amino acid residues near the glycosylation site 447:493 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 0 17 theme Molecular-scale 0:14 arg1 features 16:23 Molecular-scale features 0:23 Molecular-scale features that govern the effects of O-glycosylation on a carbohydrate-binding module 0:99 Molecular-scale features that govern the effects of O-glycosylation on a carbohydrate-binding module. 28966766 1 18 theme Protein 102:108 arg1 modification 159:170 a ubiquitous post-translational modification 127:170 a ubiquitous post-translational modification in all kingdoms of life 127:194 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 1 18 theme Protein 102:108 arg1 glycosylation 110:122 Protein glycosylation 102:122 Protein glycosylation 102:122 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 2 19 from ramifications 275:287 arg1 structure 324:332 biomolecular structure 311:332 biomolecular structure 311:332 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 2 19 from ramifications 275:287 arg1 function 338:345 function 338:345 function 338:345 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 3 20 theme acid 453:456 arg1 residues 458:465 amino acid residues 447:465 amino acid residues near the glycosylation site 447:493 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 5 21 theme multiple 859:866 arg1 properties 877:886 multiple improved properties 859:886 multiple improved properties 859:886 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 3 22 gly glycoprotein 391:402 arg1 glycoprotein 391:402 a small model glycoprotein 377:402 a small model glycoprotein 377:402 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 3 23 theme cellulose 588:596 arg1 affinity 606:613 cellulose binding affinity 588:613 cellulose binding affinity 588:613 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 5 24 theme improved 868:875 arg1 properties 877:886 multiple improved properties 859:886 multiple improved properties 859:886 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 3 25 theme binding 598:604 arg1 affinity 606:613 cellulose binding affinity 588:613 cellulose binding affinity 588:613 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 5 26 theme O-glycans 932:940 arg1 structures 918:927 the structures 914:927 the structures of O-glycans and amino acids local to the glycosylation site 914:988 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 4 27 theme features 694:701 arg1 importance 664:673 the collective importance 649:673 the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system 649:778 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 5 28 gly glycosylation 971:983 arg2 site 985:988 the glycosylation site 967:988 the glycosylation site 967:988 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 3 29 theme glycosylation 476:488 arg1 site 490:493 the glycosylation site 472:493 the glycosylation site 472:493 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 0 30 theme O-glycosylation 52:66 arg1 effects 41:47 the effects 37:47 the effects of O-glycosylation on a carbohydrate-binding module 37:99 Molecular-scale features that govern the effects of O-glycosylation on a carbohydrate-binding module. 28966766 3 31 gly glycosylation 476:488 arg2 site 490:493 the glycosylation site 472:493 the glycosylation site 472:493 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 4 32 theme O-glycosylation 749:763 arg1 effects 738:744 the most pronounced effects 718:744 the most pronounced effects of O-glycosylation in this system 718:778 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 5 33 theme designing 835:843 arg1 proteins 845:852 designing proteins 835:852 designing proteins with multiple improved properties 835:886 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 2 34 theme cellular 237:244 arg1 biology 246:252 cellular biology 237:252 cellular biology 237:252 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 1 35 theme ubiquitous 129:138 arg1 modification 159:170 a ubiquitous post-translational modification 127:170 a ubiquitous post-translational modification in all kingdoms of life 127:194 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 1 35 theme ubiquitous 129:138 arg1 glycosylation 110:122 Protein glycosylation 102:122 Protein glycosylation 102:122 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 2 36 theme O-glycosylation 292:306 arg1 ramifications 275:287 the molecular-level ramifications 255:287 the molecular-level ramifications of O-glycosylation on biomolecular structure and function 255:345 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 1 37 theme post-translational 140:157 arg1 modification 159:170 a ubiquitous post-translational modification 127:170 a ubiquitous post-translational modification in all kingdoms of life 127:194 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 1 37 theme post-translational 140:157 arg1 glycosylation 110:122 Protein glycosylation 102:122 Protein glycosylation 102:122 Protein glycosylation is a ubiquitous post-translational modification in all kingdoms of life. 28966766 5 38 with proteins 845:852 arg1 properties 877:886 multiple improved properties 859:886 multiple improved properties 859:886 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 3 39 theme glycosidic 500:509 arg1 linkage 511:517 glycosidic linkage 500:517 glycosidic linkage 500:517 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 4 40 theme study 636:640 arg1 results 620:626 The results 616:626 The results of this study 616:640 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 3 41 theme corresponding 540:552 arg1 changes 554:560 any corresponding changes 536:560 any corresponding changes to physical stability and cellulose binding affinity 536:613 Here, we took a small model glycoprotein and changed the glycan structure and size, amino acid residues near the glycosylation site, and glycosidic linkage while monitoring any corresponding changes to physical stability and cellulose binding affinity. 28966766 4 42 theme collective 653:662 arg1 importance 664:673 the collective importance 649:673 the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system 649:778 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 28966766 2 43 theme molecular-level 259:273 arg1 ramifications 275:287 the molecular-level ramifications 255:287 the molecular-level ramifications of O-glycosylation on biomolecular structure and function 255:345 Despite its importance in molecular and cellular biology, the molecular-level ramifications of O-glycosylation on biomolecular structure and function remain elusive. 28966766 5 44 theme amino 946:950 arg1 acids 952:956 amino acids 946:956 amino acids 946:956 Going forward, this study suggests the possibility of designing proteins with multiple improved properties by simultaneously varying the structures of O-glycans and amino acids local to the glycosylation site. 28966766 4 45 from effects 738:744 arg1 system 773:778 this system 768:778 this system 768:778 The results of this study reveal the collective importance of all the studied features in controlling the most pronounced effects of O-glycosylation in this system. 25753710 5 0 from JCGGDB 785:790 arg1 available 772:780 available 772:780 available 772:780 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 1 from preferences 514:524 arg1 proteins 561:568 proteins 561:568 proteins 561:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 2 gly glycosylation 538:550 arg2 sites 552:556 glycosylation sites 538:556 glycosylation sites in proteins 538:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 2 gly glycosylation 538:550 arg2 lectins 529:535 lectins 529:535 lectins 529:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 2 gly glycosylation 538:550 arg1 lectins 529:535 lectins 529:535 lectins 529:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 3 from centers 432:438 arg1 database 462:469 a glycan structure database 443:469 a glycan structure database 443:469 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 5 4 dep interfaces 702:711 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 4 dep interfaces 702:711 arg1 Search 722:727 Keyword Search 714:727 Keyword Search 714:727 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 4 dep interfaces 702:711 arg1 Search 740:745 Structure Search 730:745 Structure Search 730:745 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 4 dep interfaces 702:711 arg1 Explorer 762:769 Explorer 762:769 Explorer 762:769 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 5 theme glycan 507:512 arg1 preferences 514:524 glycan preferences 507:524 glycan preferences of lectins 507:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 6 theme related 581:587 arg1 genes 575:579 genes 575:579 genes related to glycan syntheses from glycoscience and related fields 575:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 3 7 theme cooperative 329:339 arg1 databases 341:349 5 cooperative databases 327:349 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology 327:426 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 1 8 theme biological 76:85 arg1 significance 87:98 The biological significance 72:98 The biological significance of glycans 72:109 The biological significance of glycans has been widely studied and reported in the past. 25753710 2 9 from available 224:232 arg1 databases 246:254 existing databases 237:254 existing databases 237:254 However, most achievements of our predecessors are not readily available in existing databases. 25753710 4 10 from genes 575:579 arg1 proteins 561:568 proteins 561:568 proteins 561:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 5 11 theme search 695:700 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 11 theme search 695:700 arg1 Search 722:727 Keyword Search 714:727 Keyword Search 714:727 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 11 theme search 695:700 arg1 Search 740:745 Structure Search 730:745 Structure Search 730:745 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 11 theme search 695:700 arg1 Explorer 762:769 Explorer 762:769 Explorer 762:769 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 1 12 theme glycans 103:109 arg1 significance 87:98 The biological significance 72:98 The biological significance of glycans 72:109 The biological significance of glycans has been widely studied and reported in the past. 25753710 4 13 from fields 639:644 arg1 syntheses 599:607 glycan syntheses 592:607 glycan syntheses from glycoscience and related fields 592:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 14 theme related 631:637 arg1 fields 639:644 glycoscience and related fields 614:644 glycoscience and related fields 614:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 15 from sites 552:556 arg1 proteins 561:568 proteins 561:568 proteins 561:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 2 16 theme most 170:173 arg1 achievements 175:186 most achievements 170:186 most achievements of our predecessors 170:206 However, most achievements of our predecessors are not readily available in existing databases. 25753710 3 17 from databases 341:349 arg1 AIST 318:321 AIST 318:321 AIST 318:321 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 3 17 from databases 341:349 arg1 alliance 354:361 alliance 354:361 alliance with JCGG 354:371 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 2 18 from databases 246:254 arg1 available 224:232 available 224:232 available 224:232 However, most achievements of our predecessors are not readily available in existing databases. 25753710 5 19 theme available 772:780 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 19 theme available 772:780 arg1 Search 722:727 Keyword Search 714:727 Keyword Search 714:727 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 19 theme available 772:780 arg1 Search 740:745 Structure Search 730:745 Structure Search 730:745 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 19 theme available 772:780 arg1 Explorer 762:769 Explorer 762:769 Explorer 762:769 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 20 theme lectins 529:535 arg1 lectins 529:535 lectins 529:535 lectins 529:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 20 theme lectins 529:535 arg1 genes 575:579 genes 575:579 genes related to glycan syntheses from glycoscience and related fields 575:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 20 theme lectins 529:535 arg1 sites 552:556 glycosylation sites 538:556 glycosylation sites in proteins 538:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 20 theme lectins 529:535 arg1 preferences 514:524 glycan preferences 507:524 glycan preferences of lectins 507:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 5 21 theme Keyword 714:720 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 21 theme Keyword 714:720 arg1 Search 722:727 Keyword Search 714:727 Keyword Search 714:727 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 22 theme structure 452:460 arg1 database 462:469 a glycan structure database 443:469 a glycan structure database 443:469 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 3 23 theme original 296:303 arg1 databases 305:313 15 original databases 293:313 15 original databases in AIST 293:321 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 4 24 theme glycan 445:450 arg1 database 462:469 a glycan structure database 443:469 a glycan structure database 443:469 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 0 25 dep Glycobiology 29:40 arg1 Database 62:69 Database 62:69 Database 62:69 JCGGDB: Japan Consortium for Glycobiology and Glycotechnology Database. 25753710 5 26 from available 772:780 arg1 JCGGDB 785:790 JCGGDB 785:790 JCGGDB 785:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 27 theme glycosylation 538:550 arg1 lectins 529:535 lectins 529:535 lectins 529:535 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 4 27 theme glycosylation 538:550 arg1 sites 552:556 glycosylation sites 538:556 glycosylation sites in proteins 538:568 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 3 28 dep databases 341:349 arg1 Consortium 380:389 Consortium 380:389 Consortium 380:389 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 2 29 theme predecessors 195:206 arg1 achievements 175:186 most achievements 170:186 most achievements of our predecessors 170:206 However, most achievements of our predecessors are not readily available in existing databases. 25753710 3 30 with alliance 354:361 arg1 JCGG 368:371 JCGG 368:371 JCGG 368:371 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 5 31 theme Structure 730:738 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 31 theme Structure 730:738 arg1 Search 740:745 Structure Search 730:745 Structure Search 730:745 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 32 theme major 689:693 arg1 interfaces 702:711 three major search interfaces 683:711 three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB 683:790 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 32 theme major 689:693 arg1 Search 722:727 Keyword Search 714:727 Keyword Search 714:727 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 32 theme major 689:693 arg1 Search 740:745 Structure Search 730:745 Structure Search 730:745 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 32 theme major 689:693 arg1 Explorer 762:769 Explorer 762:769 Explorer 762:769 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 5 33 theme multiple 809:816 arg1 databases 818:826 multiple databases 809:826 multiple databases 809:826 This chapter illustrates how to use three major search interfaces (Keyword Search, Structure Search, and GlycoChem Explorer) available in JCGGDB to search across multiple databases. 25753710 4 34 theme glycan 592:597 arg1 syntheses 599:607 glycan syntheses 592:607 glycan syntheses from glycoscience and related fields 592:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 2 35 theme existing 237:244 arg1 databases 246:254 existing databases 237:254 existing databases 237:254 However, most achievements of our predecessors are not readily available in existing databases. 25753710 4 36 theme glycoscience 614:625 arg1 fields 639:644 glycoscience and related fields 614:644 glycoscience and related fields 614:644 It centers on a glycan structure database and accumulates information such as glycan preferences of lectins, glycosylation sites in proteins, and genes related to glycan syntheses from glycoscience and related fields. 25753710 0 37 dep JCGGDB 0:5 arg1 Consortium 14:23 Consortium 14:23 Consortium 14:23 JCGGDB: Japan Consortium for Glycobiology and Glycotechnology Database. 25753710 3 38 from databases 305:313 arg1 AIST 318:321 AIST 318:321 AIST 318:321 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25753710 3 38 from databases 305:313 arg1 alliance 354:361 alliance 354:361 alliance with JCGG 354:371 JCGGDB is a meta-database involving 15 original databases in AIST and 5 cooperative databases in alliance with JCGG: Japan Consortium for Glycobiology and Glycotechnology. 25261856 0 0 theme lateral 93:99 arg1 sclerosis 101:109 amyotrophic lateral sclerosis 81:109 amyotrophic lateral sclerosis 81:109 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 0 1 from chain 27:31 arg1 sclerosis 101:109 amyotrophic lateral sclerosis 81:109 amyotrophic lateral sclerosis 81:109 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 2 2 with patients 453:460 arg1 diseases 473:480 other diseases 467:480 other diseases 467:480 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 5 3 theme complex 862:868 arg1 N-glycans 816:824 The N-glycans 812:824 The N-glycans found in the CSF 812:841 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 5 3 theme complex 862:868 arg1 diantennary 870:880 predominantly complex diantennary 848:880 predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage 848:923 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 6 4 dep glycans 1161:1167 arg1 A2G2S 1169:1173 A2G2S(6)1 1169:1177 A2G2S(6)1 1169:1177 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 4 dep glycans 1161:1167 arg1 FA2G2S 1183:1188 FA2G2S(3)1 1183:1192 FA2G2S(3)1 1183:1192 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 4 dep glycans 1161:1167 arg1 P=0.0348 1195:1202 P=0.0348 1195:1202 P=0.0348 1195:1202 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 4 dep glycans 1161:1167 arg1 glycans 1161:1167 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 0 5 theme amyotrophic 81:91 arg1 sclerosis 101:109 amyotrophic lateral sclerosis 81:109 amyotrophic lateral sclerosis 81:109 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 8 6 theme potential 1428:1436 arg1 change 1381:1386 this change 1376:1386 this change 1376:1386 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 8 6 theme potential 1428:1436 arg1 biomarker 1438:1446 potential biomarker 1428:1446 potential biomarker 1428:1446 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 5 7 with diantennary 870:880 arg1 acid 894:897 sialic acid 887:897 sialic acid in α2,3- and α2,6-linkage 887:923 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 1 8 theme fatal 164:168 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 8 theme fatal 164:168 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 3 9 theme mass 657:660 arg1 spectrometry 662:673 mass spectrometry 657:673 mass spectrometry 657:673 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 4 10 theme fast 794:797 arg1 progressors 799:809 fast progressors 794:809 fast progressors 794:809 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 7 11 theme pNFH 1254:1257 arg1 biomarker 1264:1272 a biomarker 1262:1272 a biomarker in ALS 1262:1279 CONCLUSIONS Our results underscore the value of pNFH as a biomarker in ALS. 25261856 7 11 theme pNFH 1254:1257 arg1 value 1245:1249 the value 1241:1249 the value of pNFH 1241:1257 CONCLUSIONS Our results underscore the value of pNFH as a biomarker in ALS. 25261856 4 12 dep RESULTS 676:682 arg1 higher 715:720 higher 715:720 higher 715:720 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 6 13 theme diantennary 1149:1159 arg1 A2G2S 1169:1173 A2G2S(6)1 1169:1177 A2G2S(6)1 1169:1177 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 13 theme diantennary 1149:1159 arg1 FA2G2S 1183:1188 FA2G2S(3)1 1183:1192 FA2G2S(3)1 1183:1192 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 13 theme diantennary 1149:1159 arg1 glycans 1161:1167 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 5 14 theme bisecting 930:938 arg1 structures 971:980 bisecting N-acetylglucosamine-containing structures 930:980 bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures 930:1027 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 2 15 dep ELISA 313:317 arg1 chain 360:364 the biomarker phosphoneurofilament heavy chain 319:364 ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 313:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 15 dep ELISA 313:317 arg1 pNFH 367:370 pNFH 367:370 pNFH 367:370 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 16 theme ALS 409:411 arg1 n=29 423:426 n=29 423:426 n=29 423:426 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 16 theme ALS 409:411 arg1 patients 413:420 ALS patients 409:420 ALS patients (n=29) 409:427 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 5 17 theme N-acetylglucosamine-containing 940:969 arg1 structures 971:980 bisecting N-acetylglucosamine-containing structures 930:980 bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures 930:1027 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 1 18 theme neurodegenerative 170:186 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 18 theme neurodegenerative 170:186 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 2 19 theme other 467:471 arg1 diseases 473:480 other diseases 467:480 other diseases 467:480 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 5 20 from acid 894:897 arg1 α2,6-linkage 912:923 α2,6-linkage 912:923 α2,6-linkage 912:923 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 5 20 from acid 894:897 arg1 α2,3- 902:906 α2,3- 902:906 α2,3- 902:906 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 5 21 gly fucosylated 1006:1016 arg1 structures 1018:1027 peripherally fucosylated structures 993:1027 bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures 930:1027 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 6 22 theme ALS 1071:1073 arg1 group 1075:1079 the ALS group 1067:1079 the ALS group 1067:1079 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 0 23 theme heavy 21:25 arg1 chain 27:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 0 24 from N-glycomics 37:47 arg1 sclerosis 101:109 amyotrophic lateral sclerosis 81:109 amyotrophic lateral sclerosis 81:109 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 2 25 theme biomarker 323:331 arg1 chain 360:364 the biomarker phosphoneurofilament heavy chain 319:364 ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 313:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 25 theme biomarker 323:331 arg1 pNFH 367:370 pNFH 367:370 pNFH 367:370 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 8 26 from variation 1311:1319 arg1 ALS 1355:1357 ALS 1355:1357 ALS 1355:1357 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 0 27 theme Phosphoneurofilament 0:19 arg1 chain 27:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 6 28 contain had 1081:1083 arg2 increase 1099:1106 a significant increase 1085:1106 a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1085:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 28 contain had 1081:1083 arg1 group 1075:1079 the ALS group 1067:1079 the ALS group 1067:1079 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 2 29 theme cerebrospinal 380:392 arg1 CSF 401:403 CSF 401:403 CSF 401:403 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 29 theme cerebrospinal 380:392 arg1 fluid 394:398 the cerebrospinal fluid 376:398 the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 376:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 4 30 with comparison 741:750 arg1 P<0.0001 767:774 P<0.0001 767:774 P<0.0001 767:774 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 4 30 with comparison 741:750 arg1 controls 757:764 controls 757:764 controls (P<0.0001) 757:775 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 1 31 theme BACKGROUND 112:121 arg1 ALS 154:156 ALS 154:156 ALS 154:156 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 31 theme BACKGROUND 112:121 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 31 theme BACKGROUND 112:121 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 5 32 theme fucosylated 1006:1016 arg1 structures 1018:1027 peripherally fucosylated structures 993:1027 bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures 930:1027 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 2 33 theme patients 413:420 arg1 CSF 401:403 CSF 401:403 CSF 401:403 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 33 theme patients 413:420 arg1 fluid 394:398 the cerebrospinal fluid 376:398 the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 376:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 6 34 theme monosialylated 1134:1147 arg1 A2G2S 1169:1173 A2G2S(6)1 1169:1177 A2G2S(6)1 1169:1177 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 34 theme monosialylated 1134:1147 arg1 FA2G2S 1183:1188 FA2G2S(3)1 1183:1192 FA2G2S(3)1 1183:1192 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 34 theme monosialylated 1134:1147 arg1 glycans 1161:1167 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 3 35 theme glycomics 603:611 arg1 approach 613:620 a glycomics approach 601:620 a glycomics approach based on liquid chromatography and mass spectrometry 601:673 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 1 36 theme Amyotrophic 123:133 arg1 ALS 154:156 ALS 154:156 ALS 154:156 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 36 theme Amyotrophic 123:133 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 36 theme Amyotrophic 123:133 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 37 theme neuron 209:214 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 37 theme neuron 209:214 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 6 38 theme peak 1113:1116 arg1 increase 1099:1106 a significant increase 1085:1106 a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1085:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 5 39 located found 826:830 arg2 diantennary 870:880 predominantly complex diantennary 848:880 predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage 848:923 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 5 39 located found 826:830 arg2 N-glycans 816:824 The N-glycans 812:824 The N-glycans found in the CSF 812:841 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 5 39 located found 826:830 arg1 CSF 839:841 the CSF 835:841 the CSF 835:841 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 3 40 theme ALS 562:564 arg1 patients 566:573 ALS patients 562:573 ALS patients 562:573 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 8 41 from pattern 1344:1350 arg1 ALS 1355:1357 ALS 1355:1357 ALS 1355:1357 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 2 42 theme patients 453:460 arg1 CSF 401:403 CSF 401:403 CSF 401:403 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 42 theme patients 453:460 arg1 fluid 394:398 the cerebrospinal fluid 376:398 the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 376:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 4 43 theme ALS 725:727 arg1 patients 729:736 ALS patients 725:736 ALS patients 725:736 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 0 44 theme cerebrospinal 58:70 arg1 fluid 72:76 the cerebrospinal fluid 54:76 the cerebrospinal fluid 54:76 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 0 45 from fluid 72:76 arg1 chain 27:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain 0:31 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 0 45 from fluid 72:76 arg1 N-glycomics 37:47 N-glycomics 37:47 N-glycomics 37:47 Phosphoneurofilament heavy chain and N-glycomics from the cerebrospinal fluid in amyotrophic lateral sclerosis. 25261856 3 46 theme CSF 555:557 arg1 N-glycosylation 532:546 protein N-glycosylation 524:546 protein N-glycosylation of the CSF in ALS patients and controls 524:586 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 2 47 with patients 413:420 arg1 diseases 473:480 other diseases 467:480 other diseases 467:480 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 1 48 theme Lateral 135:141 arg1 ALS 154:156 ALS 154:156 ALS 154:156 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 48 theme Lateral 135:141 arg1 Sclerosis 143:151 BACKGROUND Amyotrophic Lateral Sclerosis 112:151 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) 112:157 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 1 48 theme Lateral 135:141 arg1 disease 188:194 a fatal neurodegenerative disease 162:194 a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified 162:280 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 2 49 theme age-matched 433:443 arg1 n=19 483:486 n=19 483:486 n=19 483:486 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 49 theme age-matched 433:443 arg1 patients 453:460 age-matched control patients 433:460 age-matched control patients with other diseases (n=19) 433:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 8 50 theme N-glycosylation 1328:1342 arg1 pattern 1344:1350 the N-glycosylation pattern 1324:1350 the N-glycosylation pattern in ALS 1324:1357 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 3 51 theme liquid 631:636 arg1 chromatography 638:651 liquid chromatography 631:651 liquid chromatography 631:651 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 5 52 theme sialic 887:892 arg1 acid 894:897 sialic acid 887:897 sialic acid in α2,3- and α2,6-linkage 887:923 The N-glycans found in the CSF were predominantly complex diantennary with sialic acid in α2,3- and α2,6-linkage, and bisecting N-acetylglucosamine-containing structures as well as peripherally fucosylated structures were found. 25261856 4 53 theme pNFH 684:687 arg1 levels 689:694 pNFH levels 684:694 pNFH levels 684:694 RESULTS pNFH levels were significantly higher in ALS patients in comparison with controls (P<0.0001) in particular in fast progressors. 25261856 8 54 from ALS 1355:1357 arg1 variation 1311:1319 a variation 1309:1319 a variation of the N-glycosylation pattern in ALS 1309:1357 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 2 55 theme heavy 354:358 arg1 chain 360:364 the biomarker phosphoneurofilament heavy chain 319:364 ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 313:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 55 theme heavy 354:358 arg1 pNFH 367:370 pNFH 367:370 pNFH 367:370 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 3 56 theme protein 524:530 arg1 N-glycosylation 532:546 protein N-glycosylation 524:546 protein N-glycosylation of the CSF in ALS patients and controls 524:586 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 6 57 gly monosialylated 1134:1147 arg1 A2G2S 1169:1173 A2G2S(6)1 1169:1177 A2G2S(6)1 1169:1177 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 57 gly monosialylated 1134:1147 arg1 FA2G2S 1183:1188 FA2G2S(3)1 1183:1192 FA2G2S(3)1 1183:1192 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 6 57 gly monosialylated 1134:1147 arg1 glycans 1161:1167 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1130:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 7 58 dep CONCLUSIONS 1206:1216 arg1 underscore 1230:1239 underscore 1230:1239 underscore the value of pNFH as a biomarker in ALS 1230:1279 CONCLUSIONS Our results underscore the value of pNFH as a biomarker in ALS. 25261856 8 59 theme future 1410:1415 arg1 studies 1417:1423 future studies 1410:1423 future studies 1410:1423 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 2 60 theme phosphoneurofilament 333:352 arg1 chain 360:364 the biomarker phosphoneurofilament heavy chain 319:364 ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 313:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 60 theme phosphoneurofilament 333:352 arg1 pNFH 367:370 pNFH 367:370 pNFH 367:370 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 3 61 gly N-glycosylation 532:546 arg1 controls 579:586 controls 579:586 controls 579:586 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 3 61 gly N-glycosylation 532:546 arg1 CSF 555:557 the CSF 551:557 the CSF 551:557 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 3 61 gly N-glycosylation 532:546 arg1 patients 566:573 ALS patients 562:573 ALS patients 562:573 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 8 62 theme pattern 1344:1350 arg1 variation 1311:1319 a variation 1309:1319 a variation of the N-glycosylation pattern in ALS 1309:1357 In addition, we identified a variation of the N-glycosylation pattern in ALS, suggesting that this change should be explored in future studies as potential biomarker. 25261856 6 63 theme significant 1087:1097 arg1 increase 1099:1106 a significant increase 1085:1106 a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348) 1085:1203 As compared with controls the ALS group had a significant increase of a peak composed of the monosialylated diantennary glycans A2G2S(6)1 and FA2G2S(3)1 (P=0.0348). 25261856 1 64 theme motor 203:207 arg1 neuron 209:214 the motor neuron 199:214 the motor neuron 199:214 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 3 65 from N-glycosylation 532:546 arg1 controls 579:586 controls 579:586 controls 579:586 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 3 65 from N-glycosylation 532:546 arg1 patients 566:573 ALS patients 562:573 ALS patients 562:573 Furthermore, we compared protein N-glycosylation of the CSF in ALS patients and controls, by applying a glycomics approach based on liquid chromatography and mass spectrometry. 25261856 1 66 theme validated 240:248 arg1 biomarkers 250:259 no clinically validated biomarkers 226:259 no clinically validated biomarkers 226:259 BACKGROUND Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease of the motor neuron for which no clinically validated biomarkers have been identified. 25261856 2 67 theme control 445:451 arg1 n=19 483:486 n=19 483:486 n=19 483:486 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 67 theme control 445:451 arg1 patients 453:460 age-matched control patients 433:460 age-matched control patients with other diseases (n=19) 433:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 7 68 from biomarker 1264:1272 arg1 ALS 1277:1279 ALS 1277:1279 ALS 1277:1279 CONCLUSIONS Our results underscore the value of pNFH as a biomarker in ALS. 25261856 2 69 dep METHODS 283:289 arg1 quantified 299:308 quantified 299:308 have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA 294:496 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 70 from ELISA 313:317 arg1 CSF 401:403 CSF 401:403 CSF 401:403 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 25261856 2 70 from ELISA 313:317 arg1 fluid 394:398 the cerebrospinal fluid 376:398 the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) 376:487 METHODS We have quantified by ELISA the biomarker phosphoneurofilament heavy chain (pNFH) in the cerebrospinal fluid (CSF) of ALS patients (n=29) and age-matched control patients with other diseases (n=19) by ELISA. 27108103 9 0 theme strand 1295:1300 arg1 N-H 1280:1282 the N-H 1276:1282 the N-H of other β strand of the other subunit 1276:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 10 1 theme 169-Lyś 1453:1459 arg1 Asp 1449:1451 Asp 169-Lyś 163 1449:1463 Asp 169-Lyś 163 1449:1463 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 2 2 theme structure 287:295 arg1 variation 297:305 a fascinating quaternary structure variation 262:305 a fascinating quaternary structure variation 262:305 They exhibit a fascinating quaternary structure variation. 27108103 6 3 theme non-glycosylated 840:855 arg1 dimers 857:862 non-glycosylated dimers 840:862 non-glycosylated dimers 840:862 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 4 4 theme legume 406:411 arg1 agglutinin 379:388 Soybean agglutinin 371:388 Soybean agglutinin 371:388 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 4 4 theme legume 406:411 arg1 lectin 413:418 a tetrameric legume lectin 393:418 a tetrameric legume lectin 393:418 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 7 5 from temperature 927:937 arg1 degree 897:902 the high degree 888:902 the high degree of stability at normal temperature 888:937 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 2 6 theme fascinating 264:274 arg1 variation 297:305 a fascinating quaternary structure variation 262:305 a fascinating quaternary structure variation 262:305 They exhibit a fascinating quaternary structure variation. 27108103 9 7 theme subunit 1315:1321 arg1 strand 1295:1300 other β strand 1287:1300 other β strand of the other subunit 1287:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 10 8 from interface 1502:1510 arg1 Asp 1449:1451 Asp 169-Lyś 163 1449:1463 Asp 169-Lyś 163 1449:1463 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 10 8 from interface 1502:1510 arg1 Asp 1469:1471 Asp 192-Arǵ 185 1469:1483 Asp 192-Arǵ 185 1469:1483 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 10 8 from interface 1502:1510 arg1 Arg 1417:1419 Arg 185-Asṕ192 1417:1430 Arg 185-Asṕ192 1417:1430 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 10 9 theme 192-Arǵ 1473:1479 arg1 Asp 1469:1471 Asp 192-Arǵ 185 1469:1483 Asp 192-Arǵ 185 1469:1483 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 6 10 theme molecular 720:728 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 11 11 from increase 1640:1647 arg1 entropy 1664:1670 vibrational entropy 1652:1670 vibrational entropy 1652:1670 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 0 12 theme enhanced 97:104 arg1 stability 117:125 its enhanced structural stability 93:125 its enhanced structural stability 93:125 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 5 13 theme present 469:475 arg1 study 477:481 the present study 465:481 the present study 465:481 In the present study we explore the main origin for the stability of soybean agglutinin dimer. 27108103 8 14 theme folded 1108:1113 arg1 conformation 1115:1126 its overall folded conformation 1096:1126 its overall folded conformation due to high subunit interactions at the non-canonical interface 1096:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 7 15 theme ionic 976:980 arg1 ~200 kcal/mol 996:1008 ~200 kcal/mol 996:1008 ~200 kcal/mol 996:1008 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 7 15 theme ionic 976:980 arg1 interactions 982:993 interfacial ionic interactions 964:993 interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%) 964:1080 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 8 16 theme due 1128:1130 arg1 conformation 1115:1126 its overall folded conformation 1096:1126 its overall folded conformation due to high subunit interactions at the non-canonical interface 1096:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 5 17 theme agglutinin 539:548 arg1 dimer 550:554 soybean agglutinin dimer 531:554 soybean agglutinin dimer 531:554 In the present study we explore the main origin for the stability of soybean agglutinin dimer. 27108103 7 18 theme interfacial 964:974 arg1 ~200 kcal/mol 996:1008 ~200 kcal/mol 996:1008 ~200 kcal/mol 996:1008 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 7 18 theme interfacial 964:974 arg1 interactions 982:993 interfacial ionic interactions 964:993 interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%) 964:1080 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 8 19 from interface 1182:1190 arg1 interactions 1148:1159 high subunit interactions 1135:1159 high subunit interactions at the non-canonical interface 1135:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 1 20 used used 158:161 arg2 system 174:179 a model system 166:179 a model system for studying protein-carbohydrate and protein-protein interactions 166:246 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 1 20 used used 158:161 arg2 lectins 139:145 The legume lectins 128:145 The legume lectins 128:145 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 11 21 theme simulation 1607:1616 arg1 results 1618:1624 our simulation results 1603:1624 our simulation results 1603:1624 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 0 22 theme dimer 48:52 arg1 simulation 9:18 Dynamics simulation 0:18 Dynamics simulation of soybean agglutinin (SBA) dimer 0:52 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 1 23 theme legume 132:137 arg1 system 174:179 a model system 166:179 a model system for studying protein-carbohydrate and protein-protein interactions 166:246 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 1 23 theme legume 132:137 arg1 lectins 139:145 The legume lectins 128:145 The legume lectins 128:145 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 11 24 theme vibrational 1652:1662 arg1 entropy 1664:1670 vibrational entropy 1652:1670 vibrational entropy 1652:1670 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 6 25 theme glycosylation 592:604 arg1 role 584:587 the role 580:587 the role of glycosylation on the dimeric interface 580:629 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 9 26 theme hydrogen 1215:1222 arg1 bonds 1224:1228 five important hydrogen bonds 1200:1228 five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit 1200:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 6 27 dep normal 652:657 arg1 long 698:701 long 698:701 long 698:701 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 27 dep normal 652:657 arg1 298K 660:663 298K 660:663 298K 660:663 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 7 28 theme high 892:895 arg1 degree 897:902 the high degree 888:902 the high degree of stability at normal temperature 888:937 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 0 29 theme Dynamics 0:7 arg1 simulation 9:18 Dynamics simulation 0:18 Dynamics simulation of soybean agglutinin (SBA) dimer 0:52 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 10 30 theme subunit 1376:1382 arg1 interactions 1396:1407 Ten inter subunit salt-bridge interactions 1366:1407 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface 1366:1510 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 6 31 theme normal 652:657 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 10 32 theme dimensional 1557:1567 arg1 structure 1569:1577 the three dimensional structure 1547:1577 the three dimensional structure of SBA dimer 1547:1590 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 9 33 theme β 1248:1248 arg1 sheet 1250:1254 one β sheet 1244:1254 one β sheet of one subunit with the N-H of other β strand of the other subunit 1244:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 8 34 theme high 1135:1138 arg1 interactions 1148:1159 high subunit interactions 1135:1159 high subunit interactions at the non-canonical interface 1135:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 0 35 theme agglutinin 31:40 arg1 dimer 48:52 soybean agglutinin (SBA) dimer 23:52 soybean agglutinin (SBA) dimer 23:52 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 10 36 theme dimer 1586:1590 arg1 structure 1569:1577 the three dimensional structure 1547:1577 the three dimensional structure of SBA dimer 1547:1590 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 9 37 theme subunit 1263:1269 arg1 sheet 1250:1254 one β sheet 1244:1254 one β sheet of one subunit with the N-H of other β strand of the other subunit 1244:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 1 38 theme protein-carbohydrate 194:213 arg1 interactions 235:246 protein-carbohydrate and protein-protein interactions 194:246 interactions 235:246 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 11 39 theme normal 1773:1778 arg1 temperature 1780:1790 normal temperature 1773:1790 normal temperature 1773:1790 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 1 40 theme protein-protein 219:233 arg1 interactions 235:246 protein-carbohydrate and protein-protein interactions 194:246 interactions 235:246 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 9 41 theme β 1293:1293 arg1 strand 1295:1300 other β strand 1287:1300 other β strand of the other subunit 1287:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 4 42 theme tetrameric 395:404 arg1 agglutinin 379:388 Soybean agglutinin 371:388 Soybean agglutinin 371:388 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 4 42 theme tetrameric 395:404 arg1 lectin 413:418 a tetrameric legume lectin 393:418 a tetrameric legume lectin 393:418 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 6 43 theme conformational 788:801 arg1 changes 803:809 the structural and conformational changes 769:809 the structural and conformational changes between the glycosylated and non-glycosylated dimers 769:862 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 44 theme dynamics 730:737 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 45 theme MD 740:741 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 2 46 theme quaternary 276:285 arg1 variation 297:305 a fascinating quaternary structure variation 262:305 a fascinating quaternary structure variation 262:305 They exhibit a fascinating quaternary structure variation. 27108103 9 47 theme other 1309:1313 arg1 subunit 1315:1321 the other subunit 1305:1321 the other subunit 1305:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 4 48 theme Soybean 371:377 arg1 agglutinin 379:388 Soybean agglutinin 371:388 Soybean agglutinin 371:388 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 4 48 theme Soybean 371:377 arg1 lectin 413:418 a tetrameric legume lectin 393:418 a tetrameric legume lectin 393:418 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 9 49 with sheet 1250:1254 arg1 N-H 1280:1282 the N-H 1276:1282 the N-H of other β strand of the other subunit 1276:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 5 50 theme main 498:501 arg1 origin 503:508 the main origin 494:508 the main origin for the stability of soybean agglutinin dimer 494:554 In the present study we explore the main origin for the stability of soybean agglutinin dimer. 27108103 8 51 theme subunit 1140:1146 arg1 interactions 1148:1159 high subunit interactions 1135:1159 high subunit interactions at the non-canonical interface 1135:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 11 52 theme free 1691:1694 arg1 energy 1696:1701 the free energy 1687:1701 the free energy 1687:1701 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 0 53 theme structural 106:115 arg1 stability 117:125 its enhanced structural stability 93:125 its enhanced structural stability 93:125 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 6 54 theme solvent 712:718 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 1 55 theme model 168:172 arg1 system 174:179 a model system 166:179 a model system for studying protein-carbohydrate and protein-protein interactions 166:246 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 1 55 theme model 168:172 arg1 lectins 139:145 The legume lectins 128:145 The legume lectins 128:145 The legume lectins are widely used as a model system for studying protein-carbohydrate and protein-protein interactions. 27108103 0 56 from impact 66:71 arg1 stability 117:125 its enhanced structural stability 93:125 its enhanced structural stability 93:125 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 7 57 theme polar 1019:1023 arg1 residues 1025:1032 polar residues 1019:1032 polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%) 1019:1080 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 9 58 theme structural 1344:1353 arg1 integrity 1355:1363 the structural integrity 1340:1363 the structural integrity 1340:1363 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 8 59 theme overall 1100:1106 arg1 conformation 1115:1126 its overall folded conformation 1096:1126 its overall folded conformation due to high subunit interactions at the non-canonical interface 1096:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 8 60 theme non-canonical 1168:1180 arg1 interface 1182:1190 the non-canonical interface 1164:1190 the non-canonical interface 1164:1190 It maintains its overall folded conformation due to high subunit interactions at the non-canonical interface. 27108103 10 61 theme non-canonical 1488:1500 arg1 interface 1502:1510 non-canonical interface 1488:1510 non-canonical interface 1488:1510 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 5 62 theme soybean 531:537 arg1 dimer 550:554 soybean agglutinin dimer 531:554 soybean agglutinin dimer 531:554 In the present study we explore the main origin for the stability of soybean agglutinin dimer. 27108103 6 63 dep understand 569:578 arg1 to 566:567 to 566:567 to 566:567 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 4 64 theme each 421:424 arg1 subunits 435:442 subunits 435:442 subunits 435:442 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 6 65 gly non-glycosylated 840:855 arg1 dimers 857:862 non-glycosylated dimers 840:862 non-glycosylated dimers 840:862 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 5 66 theme dimer 550:554 arg1 stability 518:526 the stability 514:526 the stability of soybean agglutinin dimer 514:554 In the present study we explore the main origin for the stability of soybean agglutinin dimer. 27108103 6 67 theme dimeric 613:619 arg1 interface 621:629 the dimeric interface 609:629 the dimeric interface 609:629 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 0 68 theme glycosylation 76:88 arg1 impact 66:71 the impact 62:71 the impact of glycosylation on its enhanced structural stability 62:125 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 9 69 theme important 1205:1213 arg1 bonds 1224:1228 five important hydrogen bonds 1200:1228 five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit 1200:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 7 70 theme normal 920:925 arg1 temperature 927:937 normal temperature 920:937 normal temperature 920:937 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 6 71 theme high 667:670 arg1 temperatures 672:683 high temperatures 667:683 high temperatures (380K, 500K) 667:696 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 7 72 theme stability 907:915 arg1 degree 897:902 the high degree 888:902 the high degree of stability at normal temperature 888:937 The study reveals that the high degree of stability at normal temperature is mostly contributed by interfacial ionic interactions (~200 kcal/mol) between polar residues like Lys, Arg, Asp, Thr, Ser, Asn and Gln (62%). 27108103 0 73 theme soybean 23:29 arg1 SBA 43:45 SBA 43:45 SBA 43:45 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 0 73 theme soybean 23:29 arg1 agglutinin 31:40 soybean agglutinin 23:40 soybean agglutinin (SBA) dimer 23:52 Dynamics simulation of soybean agglutinin (SBA) dimer reveals the impact of glycosylation on its enhanced structural stability. 27108103 11 74 from temperature 1780:1790 arg1 stabilization 1740:1752 the glycan-induced stabilization 1721:1752 the glycan-induced stabilization by ~45 kcal/mol at normal temperature 1721:1790 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 10 75 theme inter 1370:1374 arg1 interactions 1396:1407 Ten inter subunit salt-bridge interactions 1366:1407 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface 1366:1510 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 9 76 theme sheet 1250:1254 arg1 CO 1238:1239 CO 1238:1239 CO of one β sheet of one subunit with the N-H of other β strand of the other subunit 1238:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27108103 6 77 theme explicit 703:710 arg1 simulations 744:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations 652:754 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 78 from role 584:587 arg1 interface 621:629 the dimeric interface 609:629 the dimeric interface 609:629 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 10 79 theme SBA 1582:1584 arg1 dimer 1586:1590 SBA dimer 1582:1590 SBA dimer 1582:1590 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 6 80 theme structural 773:782 arg1 changes 803:809 the structural and conformational changes 769:809 the structural and conformational changes between the glycosylated and non-glycosylated dimers 769:862 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 81 dep temperatures 672:683 arg1 380K 686:689 380K 686:689 380K 686:689 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 6 81 dep temperatures 672:683 arg1 500K 692:695 500K 692:695 500K 692:695 In order to understand the role of glycosylation on the dimeric interface, we have carried out normal (298K), high temperatures (380K, 500K) long explicit solvent molecular dynamics (MD) simulations and compared the structural and conformational changes between the glycosylated and non-glycosylated dimers. 27108103 10 82 theme salt-bridge 1384:1394 arg1 interactions 1396:1407 Ten inter subunit salt-bridge interactions 1366:1407 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface 1366:1510 Ten inter subunit salt-bridge interactions between Arg 185-Asṕ192, Lys 163-Asṕ169, Asp 169-Lyś 163 and Asp 192-Arǵ 185 at non-canonical interface appear to be important to maintain the three dimensional structure of SBA dimer. 27108103 11 83 theme glycan-induced 1725:1738 arg1 stabilization 1740:1752 the glycan-induced stabilization 1721:1752 the glycan-induced stabilization by ~45 kcal/mol at normal temperature 1721:1790 Moreover, our simulation results revealed that increase in vibrational entropy could decrease the free energy and contribute to the glycan-induced stabilization by ~45 kcal/mol at normal temperature. 27108103 4 84 gly glycosylated 448:459 arg1 subunits 435:442 subunits 435:442 subunits 435:442 Soybean agglutinin is a tetrameric legume lectin, each of whose subunits are glycosylated. 27108103 9 85 theme other 1287:1291 arg1 strand 1295:1300 other β strand 1287:1300 other β strand of the other subunit 1287:1321 Mainly five important hydrogen bonds between CO of one β sheet of one subunit with the N-H of other β strand of the other subunit help to maintain the structural integrity. 27867113 6 0 with contrast 1154:1161 arg1 structures 1190:1199 smaller and truncated structures 1168:1199 smaller and truncated structures among IFN4NHEK glycans 1168:1222 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 9 1 theme cell 1708:1711 arg1 lines 1713:1717 different tumor-derived cell lines 1684:1717 different tumor-derived cell lines 1684:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 11 2 theme HEK293 1917:1922 arg1 cells 1924:1928 HEK293 cells 1917:1928 HEK293 cells 1917:1928 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 8 3 theme plasma 1419:1424 arg1 concentration 1426:1438 maximum plasma concentration 1411:1438 maximum plasma concentration 3-times faster than IFN4NCHO 1411:1467 In fact, although IFN4NHEK reached maximum plasma concentration 3-times faster than IFN4NCHO, their elimination profile did not differ significantly. 27867113 3 4 theme cell 577:580 arg1 lines 582:586 CHO and HEK cell lines 565:586 lines 582:586 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 1 5 theme HEK 127:129 arg1 CHO 119:121 Both CHO and HEK cells 114:135 CHO 119:121 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 1 5 theme HEK 127:129 arg1 cells 131:135 Both CHO and HEK cells 114:135 cells 131:135 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 1 5 theme HEK 127:129 arg1 hosts 153:157 interesting hosts 141:157 interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation 141:284 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 7 6 from impact 1320:1325 arg1 rate 1344:1347 the clearance rate 1330:1347 the clearance rate of both molecules in rats 1330:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 2 7 theme oligosaccharide 299:313 arg1 structures 315:324 oligosaccharide structures 299:324 oligosaccharide structures attached to proteins 299:345 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 1 8 theme biotherapeutics 181:195 arg1 production 167:176 the production 163:176 the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation 163:284 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 10 9 from activity 1776:1783 arg1 models 1795:1800 animal models 1788:1800 animal models 1788:1800 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 27867113 6 10 theme IFN4NHEK 1207:1214 arg1 glycans 1216:1222 IFN4NHEK glycans 1207:1222 IFN4NHEK glycans 1207:1222 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 11 11 theme selection 1858:1866 arg1 importance 1824:1833 the importance 1820:1833 the importance of an appropriate host selection to set 1820:1873 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 3 12 theme extensive 595:603 arg1 characterization 605:620 an extensive characterization 592:620 an extensive characterization of their properties 592:640 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 7 13 from rate 1344:1347 arg1 rats 1370:1373 rats 1370:1373 rats 1370:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 10 14 dep higher 1751:1756 arg1 in 1758:1759 in 1758:1759 in 1758:1759 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 27867113 11 15 theme appropriate 1841:1851 arg1 selection 1858:1866 an appropriate host selection 1838:1866 an appropriate host selection 1838:1866 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 4 16 theme average 685:691 arg1 mass 703:706 a higher average molecular mass 676:706 a higher average molecular mass 676:706 IFN4NCHO exhibited a higher average molecular mass and more acidic isoforms compared to IFN4NHEK. 27867113 3 17 theme properties 631:640 arg1 characterization 605:620 an extensive characterization 592:620 an extensive characterization of their properties 592:640 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 6 18 theme WAX 984:986 arg1 chromatography 988:1001 WAX chromatography 984:1001 WAX chromatography 984:1001 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 5 19 with agreement 758:766 arg1 results 779:785 these results 773:785 these results 773:785 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 9 20 theme specific 1563:1570 arg1 activity 1583:1590 the in vitro antiviral specific biological activity 1540:1590 the in vitro antiviral specific biological activity of both proteins 1540:1607 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 20 theme specific 1563:1570 arg1 same 1617:1620 same 1617:1620 same 1617:1620 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 0 21 theme HEK293 84:89 arg1 cells 91:95 HEK293 cells 84:95 HEK293 cells 84:95 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 5 22 theme sialic 805:810 arg1 content 817:823 a 2-times higher sialic acid content 788:823 a 2-times higher sialic acid content 788:823 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 6 23 theme mass 1024:1027 arg1 spectrometry 1029:1040 mass spectrometry 1024:1040 mass spectrometry 1024:1040 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 2 24 theme human 471:475 arg1 cells 485:489 human derived cells 471:489 human derived cells 471:489 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 9 25 dep in 1544:1545 arg1 vitro 1547:1551 vitro 1547:1551 vitro 1547:1551 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 7 26 theme remarkable 1245:1254 arg1 differences 1256:1266 those remarkable differences 1239:1266 those remarkable differences in the glycosylation pattern 1239:1295 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 0 27 theme hyperglycosylated 48:64 arg1 IFN-α2 66:71 hyperglycosylated IFN-α2 48:71 hyperglycosylated IFN-α2 48:71 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 9 28 from agent 1675:1679 arg1 lines 1713:1717 different tumor-derived cell lines 1684:1717 different tumor-derived cell lines 1684:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 6 29 theme structures 1118:1127 arg1 existence 1069:1077 the existence 1065:1077 the existence of highly sialylated and also branched structures for IFN4NCHO glycans 1065:1148 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 1 30 theme interesting 141:151 arg1 CHO 119:121 Both CHO and HEK cells 114:135 CHO 119:121 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 1 30 theme interesting 141:151 arg1 cells 131:135 Both CHO and HEK cells 114:135 cells 131:135 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 1 30 theme interesting 141:151 arg1 hosts 153:157 interesting hosts 141:157 interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation 141:284 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 10 31 dep in 1758:1759 arg1 vivo 1761:1764 vivo 1761:1764 vivo 1761:1764 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 27867113 5 32 located found 829:833 arg1 comparison 851:860 comparison 851:860 comparison with the HEK-derived protein 851:889 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 5 32 located found 829:833 arg2 content 817:823 a 2-times higher sialic acid content 788:823 a 2-times higher sialic acid content 788:823 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 5 32 located found 829:833 arg1 agreement 758:766 agreement 758:766 agreement with these results 758:785 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 2 33 theme derived 477:483 arg1 cells 485:489 human derived cells 471:489 human derived cells 471:489 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 0 34 theme antiproliferative 18:34 arg1 activity 36:43 antiproliferative activity 18:43 antiproliferative activity 18:43 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 6 35 theme sialylated 1089:1098 arg1 existence 1069:1077 the existence 1065:1077 the existence of highly sialylated and also branched structures for IFN4NCHO glycans 1065:1148 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 3 36 gly hyperglycosylated 508:524 arg1 IFN4N 542:546 IFN4N 542:546 IFN4N 542:546 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 3 36 gly hyperglycosylated 508:524 arg1 mutein 534:539 a hyperglycosylated IFN-α2b mutein 506:539 a hyperglycosylated IFN-α2b mutein (IFN4N) 506:547 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 10 37 theme antitumor 1766:1774 arg1 activity 1776:1783 a higher in vivo antitumor activity 1749:1783 a higher in vivo antitumor activity in animal models 1749:1800 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 27867113 0 38 theme IFN-α2 66:71 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation 0:12 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 0 38 theme IFN-α2 66:71 arg1 activity 36:43 antiproliferative activity 18:43 antiproliferative activity 18:43 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 9 39 theme antiproliferative 1657:1673 arg1 agent 1675:1679 an antiproliferative agent 1654:1679 an antiproliferative agent in different tumor-derived cell lines 1654:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 7 40 from molecules 1357:1365 arg1 rats 1370:1373 rats 1370:1373 rats 1370:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 6 41 theme smaller 1168:1174 arg1 structures 1190:1199 smaller and truncated structures 1168:1199 smaller and truncated structures among IFN4NHEK glycans 1168:1222 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 8 42 theme elimination 1476:1486 arg1 profile 1488:1494 their elimination profile 1470:1494 their elimination profile 1470:1494 In fact, although IFN4NHEK reached maximum plasma concentration 3-times faster than IFN4NCHO, their elimination profile did not differ significantly. 27867113 3 43 theme IFN-α2b 526:532 arg1 IFN4N 542:546 IFN4N 542:546 IFN4N 542:546 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 3 43 theme IFN-α2b 526:532 arg1 mutein 534:539 a hyperglycosylated IFN-α2b mutein 506:539 a hyperglycosylated IFN-α2b mutein (IFN4N) 506:547 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 1 44 theme post-translational 231:248 arg1 modifications 250:262 post-translational modifications 231:262 post-translational modifications such as glycosylation 231:284 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 1 44 theme post-translational 231:248 arg1 glycosylation 272:284 glycosylation 272:284 glycosylation 272:284 Both CHO and HEK cells are interesting hosts for the production of biotherapeutics due to their ability to introduce post-translational modifications such as glycosylation. 27867113 0 45 gly hyperglycosylated 48:64 arg1 IFN-α2 66:71 hyperglycosylated IFN-α2 48:71 hyperglycosylated IFN-α2 48:71 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 7 46 from rats 1370:1373 arg1 rate 1344:1347 the clearance rate 1330:1347 the clearance rate of both molecules in rats 1330:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 11 47 theme cells 1924:1928 arg1 use 1910:1912 the use 1906:1912 the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical 1906:2003 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 9 48 theme tumor-derived 1694:1706 arg1 lines 1713:1717 different tumor-derived cell lines 1684:1717 different tumor-derived cell lines 1684:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 6 49 theme IFN4NCHO 1133:1140 arg1 glycans 1142:1148 IFN4NCHO glycans 1133:1148 IFN4NCHO glycans 1133:1148 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 3 50 theme HEK 573:575 arg1 lines 582:586 CHO and HEK cell lines 565:586 lines 582:586 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 6 51 theme monosaccharide 926:939 arg1 quantification 941:954 monosaccharide quantification 926:954 monosaccharide quantification 926:954 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 3 52 theme CHO 565:567 arg1 lines 582:586 CHO and HEK cell lines 565:586 lines 582:586 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 4 53 theme molecular 693:701 arg1 mass 703:706 a higher average molecular mass 676:706 a higher average molecular mass 676:706 IFN4NCHO exhibited a higher average molecular mass and more acidic isoforms compared to IFN4NHEK. 27867113 7 54 theme glycosylation 1275:1287 arg1 pattern 1289:1295 the glycosylation pattern 1271:1295 the glycosylation pattern 1271:1295 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 9 55 link tumor-derived 1694:1706 arg1 lines 1713:1717 different tumor-derived cell lines 1684:1717 different tumor-derived cell lines 1684:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 5 56 with comparison 851:860 arg1 protein 883:889 the HEK-derived protein 867:889 the HEK-derived protein 867:889 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 6 57 theme truncated 1180:1188 arg1 structures 1190:1199 smaller and truncated structures 1168:1199 smaller and truncated structures among IFN4NHEK glycans 1168:1222 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 11 58 theme host 1853:1856 arg1 selection 1858:1866 an appropriate host selection 1838:1866 an appropriate host selection 1838:1866 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 5 59 theme HEK-derived 871:881 arg1 protein 883:889 the HEK-derived protein 867:889 the HEK-derived protein 867:889 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 2 60 attach attached 326:333 arg1 proteins 338:345 proteins 338:345 proteins 338:345 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 2 60 attach attached 326:333 arg2 structures 315:324 oligosaccharide structures 299:324 oligosaccharide structures attached to proteins 299:345 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 7 61 from differences 1256:1266 arg1 pattern 1289:1295 the glycosylation pattern 1271:1295 the glycosylation pattern 1271:1295 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 4 62 theme higher 678:683 arg1 mass 703:706 a higher average molecular mass 676:706 a higher average molecular mass 676:706 IFN4NCHO exhibited a higher average molecular mass and more acidic isoforms compared to IFN4NHEK. 27867113 9 63 theme in 1544:1545 arg1 activity 1583:1590 the in vitro antiviral specific biological activity 1540:1590 the in vitro antiviral specific biological activity of both proteins 1540:1607 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 63 theme in 1544:1545 arg1 same 1617:1620 same 1617:1620 same 1617:1620 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 64 theme antiviral 1553:1561 arg1 activity 1583:1590 the in vitro antiviral specific biological activity 1540:1590 the in vitro antiviral specific biological activity of both proteins 1540:1607 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 64 theme antiviral 1553:1561 arg1 same 1617:1620 same 1617:1620 same 1617:1620 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 65 theme biological 1572:1581 arg1 activity 1583:1590 the in vitro antiviral specific biological activity 1540:1590 the in vitro antiviral specific biological activity of both proteins 1540:1607 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 65 theme biological 1572:1581 arg1 same 1617:1620 same 1617:1620 same 1617:1620 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 11 66 theme pharmaceutical 1990:2003 arg1 production 1938:1947 the production 1934:1947 the production of a new hyperglycosylated protein-based pharmaceutical 1934:2003 Our results show the importance of an appropriate host selection to set up a bioprocess and potentiate the use of HEK293 cells for the production of a new hyperglycosylated protein-based pharmaceutical. 27867113 4 67 theme acidic 717:722 arg1 isoforms 724:731 more acidic isoforms 712:731 more acidic isoforms 712:731 IFN4NCHO exhibited a higher average molecular mass and more acidic isoforms compared to IFN4NHEK. 27867113 5 68 theme higher 798:803 arg1 content 817:823 a 2-times higher sialic acid content 788:823 a 2-times higher sialic acid content 788:823 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 5 69 link HEK-derived 871:881 arg1 protein 883:889 the HEK-derived protein 867:889 the HEK-derived protein 867:889 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 9 70 theme proteins 1600:1607 arg1 activity 1583:1590 the in vitro antiviral specific biological activity 1540:1590 the in vitro antiviral specific biological activity of both proteins 1540:1607 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 9 70 theme proteins 1600:1607 arg1 same 1617:1620 same 1617:1620 same 1617:1620 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 6 71 theme branched 1109:1116 arg1 structures 1118:1127 also branched structures 1104:1127 also branched structures 1104:1127 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 2 72 gly glycoproteins 432:444 arg1 glycoproteins 432:444 therapeutic glycoproteins 420:444 therapeutic glycoproteins expressed in hamster and human derived cells 420:489 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 8 73 theme maximum 1411:1417 arg1 concentration 1426:1438 maximum plasma concentration 1411:1438 maximum plasma concentration 3-times faster than IFN4NCHO 1411:1467 In fact, although IFN4NHEK reached maximum plasma concentration 3-times faster than IFN4NCHO, their elimination profile did not differ significantly. 27867113 2 74 theme therapeutic 420:430 arg1 glycoproteins 432:444 therapeutic glycoproteins 420:444 therapeutic glycoproteins expressed in hamster and human derived cells 420:489 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 6 75 with agreement 911:919 arg1 quantification 941:954 monosaccharide quantification 926:954 monosaccharide quantification 926:954 This result was in agreement with monosaccharide quantification and glycan's analysis using WAX chromatography and HILIC coupled to mass spectrometry; all methods supported the existence of highly sialylated and also branched structures for IFN4NCHO glycans, in contrast with smaller and truncated structures among IFN4NHEK glycans. 27867113 7 76 theme molecules 1357:1365 arg1 rate 1344:1347 the clearance rate 1330:1347 the clearance rate of both molecules in rats 1330:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 3 77 theme hyperglycosylated 508:524 arg1 IFN4N 542:546 IFN4N 542:546 IFN4N 542:546 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 3 77 theme hyperglycosylated 508:524 arg1 mutein 534:539 a hyperglycosylated IFN-α2b mutein 506:539 a hyperglycosylated IFN-α2b mutein (IFN4N) 506:547 In this work, a hyperglycosylated IFN-α2b mutein (IFN4N) was produced in CHO and HEK cell lines and an extensive characterization of their properties was performed. 27867113 5 78 theme acid 812:815 arg1 content 817:823 a 2-times higher sialic acid content 788:823 a 2-times higher sialic acid content 788:823 In agreement with these results, a 2-times higher sialic acid content was found for IFN4NCHO in comparison with the HEK-derived protein. 27867113 0 79 gly Glycosylation 0:12 arg1 IFN-α2 66:71 hyperglycosylated IFN-α2 48:71 hyperglycosylated IFN-α2 48:71 Glycosylation and antiproliferative activity of hyperglycosylated IFN-α2 potentiate HEK293 cells as biofactories. 27867113 8 80 theme faster 1448:1453 arg1 concentration 1426:1438 maximum plasma concentration 1411:1438 maximum plasma concentration 3-times faster than IFN4NCHO 1411:1467 In fact, although IFN4NHEK reached maximum plasma concentration 3-times faster than IFN4NCHO, their elimination profile did not differ significantly. 27867113 7 81 theme clearance 1334:1342 arg1 rate 1344:1347 the clearance rate 1330:1347 the clearance rate of both molecules in rats 1330:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 2 82 theme many 379:382 arg1 differences 384:394 many differences 379:394 many differences 379:394 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 2 83 link derived 477:483 arg1 cells 485:489 human derived cells 471:489 human derived cells 471:489 Even though oligosaccharide structures attached to proteins are conserved among eukaryotes, many differences have been found between therapeutic glycoproteins expressed in hamster and human derived cells. 27867113 10 84 theme higher 1751:1756 arg1 activity 1776:1783 a higher in vivo antitumor activity 1749:1783 a higher in vivo antitumor activity in animal models 1749:1800 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 27867113 9 85 theme different 1684:1692 arg1 lines 1713:1717 different tumor-derived cell lines 1684:1717 different tumor-derived cell lines 1684:1717 Also, despite the in vitro antiviral specific biological activity of both proteins was the same, IFN4NHEK was more efficient as an antiproliferative agent in different tumor-derived cell lines. 27867113 7 86 theme considerable 1307:1318 arg1 impact 1320:1325 a considerable impact 1305:1325 a considerable impact on the clearance rate of both molecules in rats 1305:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 7 87 contain had 1297:1299 arg1 differences 1256:1266 those remarkable differences 1239:1266 those remarkable differences in the glycosylation pattern 1239:1295 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 7 87 contain had 1297:1299 arg2 impact 1320:1325 a considerable impact 1305:1325 a considerable impact on the clearance rate of both molecules in rats 1305:1373 Unexpectedly, those remarkable differences in the glycosylation pattern had not a considerable impact on the clearance rate of both molecules in rats. 27867113 10 88 theme animal 1788:1793 arg1 models 1795:1800 animal models 1788:1800 animal models 1788:1800 Accordingly, IFN4NHEK showed a higher in vivo antitumor activity in animal models. 28582506 6 0 theme most 1164:1167 arg1 types 1175:1179 most sugar types 1164:1179 most sugar types 1164:1179 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 3 1 theme different 540:548 arg1 positions 550:558 different positions 540:558 different positions 540:558 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 9 2 theme intuitive 1631:1639 arg1 tool 1664:1667 an intuitive glycan sequence search tool 1628:1667 an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB 1628:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 6 3 theme glycoconjugates 1143:1157 arg1 input 1126:1130 input 1126:1130 input of various glycoconjugates with most sugar types and chemical modifications 1126:1206 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 6 3 theme glycoconjugates 1143:1157 arg1 system 1115:1120 the simulation system 1100:1120 the simulation system 1100:1120 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 4 4 theme exact 651:655 arg1 annotations 657:667 exact annotations 651:667 exact annotations for most carbohydrate derivatives 651:701 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 2 5 theme Protein 254:260 arg1 Bank 267:270 Protein Data Bank 254:270 Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 254:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 5 theme Protein 254:260 arg1 PDB 273:275 PDB 273:275 PDB 273:275 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 1 6 theme many 151:154 arg1 processes 177:185 many essential biological processes 151:185 many essential biological processes 151:185 MOTIVATION Glycans play a central role in many essential biological processes. 28582506 9 7 theme sequence 1648:1655 arg1 tool 1664:1667 an intuitive glycan sequence search tool 1628:1667 an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB 1628:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 7 8 theme states 1408:1413 arg1 addition/deletion/modification 1282:1311 addition/deletion/modification 1282:1311 addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1282:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 2 9 theme files 278:282 arg1 reading 243:249 the reading 239:249 the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 239:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 1 10 theme biological 166:175 arg1 processes 177:185 many essential biological processes 151:185 many essential biological processes 151:185 MOTIVATION Glycans play a central role in many essential biological processes. 28582506 9 11 theme complex 1673:1679 arg1 structures 1688:1697 complex glycan structures 1673:1697 complex glycan structures with various chemical modifications in the PDB 1673:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 8 12 theme input 1442:1446 arg1 files 1448:1452 The simulation system and input files 1416:1452 files 1448:1452 The simulation system and input files can be used for CHARMM, NAMD, GROMACS, AMBER, GENESIS, LAMMPS, Desmond, OpenMM, and CHARMM/OpenMM. 28582506 7 13 theme linkages 1385:1392 arg1 addition/deletion/modification 1282:1311 addition/deletion/modification 1282:1311 addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1282:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 3 14 theme diverse 594:600 arg1 space 583:587 their chemical space 568:587 their chemical space much diverse 568:600 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 10 15 dep AVAILABILITY 1747:1758 arg1 //www.charmm-gui.org/input/glycan 1784:1816 //www.charmm-gui.org/input/glycan 1784:1816 //www.charmm-gui.org/input/glycan 1784:1816 AVAILABILITY AND IMPLEMENTATION http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. 28582506 10 15 dep AVAILABILITY 1747:1758 arg1 http 1779:1782 http 1779:1782 http 1779:1782 AVAILABILITY AND IMPLEMENTATION http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. 28582506 10 15 dep AVAILABILITY 1747:1758 arg1 //www.glycanstructure.org 1827:1851 //www.glycanstructure.org 1827:1851 AVAILABILITY AND IMPLEMENTATION http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. 1747:1852 AVAILABILITY AND IMPLEMENTATION http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. 28582506 10 15 dep AVAILABILITY 1747:1758 arg1 http 1822:1825 http 1822:1825 http 1822:1825 AVAILABILITY AND IMPLEMENTATION http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. 28582506 8 16 used used 1461:1464 arg2 system 1431:1436 The simulation system and input files 1416:1452 system 1431:1436 The simulation system and input files can be used for CHARMM, NAMD, GROMACS, AMBER, GENESIS, LAMMPS, Desmond, OpenMM, and CHARMM/OpenMM. 28582506 8 16 used used 1461:1464 arg2 files 1448:1452 The simulation system and input files 1416:1452 files 1448:1452 The simulation system and input files can be used for CHARMM, NAMD, GROMACS, AMBER, GENESIS, LAMMPS, Desmond, OpenMM, and CHARMM/OpenMM. 28582506 5 17 theme PDB 1010:1012 arg1 formats 1029:1035 both PDB and PDBx/mmCIF formats 1005:1035 formats 1029:1035 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 9 18 theme various 1704:1710 arg1 modifications 1721:1733 various chemical modifications 1704:1733 various chemical modifications in the PDB 1704:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 7 19 theme chemical 1350:1357 arg1 modifications 1359:1371 chemical modifications 1350:1371 chemical modifications 1350:1371 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 6 20 theme chemical 1185:1192 arg1 modifications 1194:1206 chemical modifications 1185:1206 chemical modifications 1185:1206 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 5 21 dep modifications 943:955 arg1 including 958:966 including 958:966 including various glycolipids 958:986 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 5 22 theme PDBx/mmCIF 1018:1027 arg1 formats 1029:1035 both PDB and PDBx/mmCIF formats 1005:1035 formats 1029:1035 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 5 23 theme most 913:916 arg1 types 924:928 most sugar types 913:928 most sugar types 913:928 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 7 24 theme types 1343:1347 arg1 addition/deletion/modification 1282:1311 addition/deletion/modification 1282:1311 addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1282:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 4 25 theme carbohydrate 678:689 arg1 derivatives 691:701 most carbohydrate derivatives 673:701 most carbohydrate derivatives 673:701 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 2 26 theme connectivity 457:468 arg1 information 470:480 connectivity information 457:480 connectivity information 457:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 0 27 theme chemical 60:67 arg1 modifications 69:81 chemical modifications 60:81 chemical modifications 60:81 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 12 28 theme Bioinformatics 1943:1956 arg1 online 1958:1963 Bioinformatics online 1943:1963 Bioinformatics online 1943:1963 SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 28582506 3 29 contain have 501:504 arg2 modifications 523:535 various chemical modifications 506:535 various chemical modifications 506:535 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 3 29 contain have 501:504 arg1 Carbohydrates 483:495 Carbohydrates 483:495 Carbohydrates 483:495 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 2 30 theme Bank 267:270 arg1 files 278:282 Protein Data Bank (PDB) files 254:282 Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 254:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 31 theme linkages 375:382 arg1 annotation 339:348 annotation 339:348 annotation 339:348 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 31 theme linkages 375:382 arg1 detection 325:333 automatic detection 315:333 automatic detection 315:333 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 0 32 theme Protein 90:96 arg1 Bank 103:106 the Protein Data Bank 86:106 the Protein Data Bank 86:106 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 5 33 theme various 968:974 arg1 glycolipids 976:986 various glycolipids 968:986 various glycolipids 968:986 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 7 34 theme sugar 1337:1341 arg1 types 1343:1347 sugar types 1337:1347 sugar types 1337:1347 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 4 35 theme PDB 724:726 arg1 chains 735:740 PDB glycan chains 724:740 PDB glycan chains 724:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 9 36 with structures 1688:1697 arg1 modifications 1721:1733 various chemical modifications 1704:1733 various chemical modifications in the PDB 1704:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 0 37 theme Glycan 0:5 arg1 Reader 7:12 Glycan Reader 0:12 Glycan Reader 0:12 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 7 38 theme types 1330:1334 arg1 addition/deletion/modification 1282:1311 addition/deletion/modification 1282:1311 addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1282:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 4 39 theme chains 735:740 arg1 chains 735:740 PDB glycan chains 724:740 PDB glycan chains 724:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 4 39 theme chains 735:740 arg1 % 719:719 more than 50% 707:719 more than 50% of PDB glycan chains 707:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 3 40 theme chemical 514:521 arg1 modifications 523:535 various chemical modifications 506:535 various chemical modifications 506:535 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 5 41 theme chemical 934:941 arg1 modifications 943:955 chemical modifications 934:955 chemical modifications (including various glycolipids) 934:987 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 2 42 theme sugar 392:396 arg1 units 398:402 sugar units 392:402 sugar units 392:402 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 12 43 theme INFORMATION 1895:1905 arg1 data 1921:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 28582506 9 44 theme Glycan 1553:1558 arg1 Database 1569:1576 Glycan Fragment Database 1553:1576 Glycan Fragment Database in GlycanStructure.Org 1553:1599 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 2 45 theme automatic 315:323 arg1 detection 325:333 automatic detection 315:333 automatic detection 315:333 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 0 46 theme most 39:42 arg1 types 50:54 most sugar types 39:54 most sugar types 39:54 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 7 47 theme glycan 1256:1261 arg1 structures 1263:1272 the glycan structures 1252:1272 the glycan structures 1252:1272 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 2 48 theme Glycan 188:193 arg1 Reader 195:200 Glycan Reader 188:200 Glycan Reader 188:200 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 8 49 theme simulation 1420:1429 arg1 system 1431:1436 The simulation system and input files 1416:1452 system 1431:1436 The simulation system and input files can be used for CHARMM, NAMD, GROMACS, AMBER, GENESIS, LAMMPS, Desmond, OpenMM, and CHARMM/OpenMM. 28582506 11 50 theme CONTACT 1854:1860 arg1 lehigh.edu 1869:1878 CONTACT wonpil@lehigh.edu 1854:1878 CONTACT wonpil@lehigh.edu. 1854:1879 CONTACT wonpil@lehigh.edu. 28582506 9 51 from modifications 1721:1733 arg1 PDB 1742:1744 the PDB 1738:1744 the PDB 1738:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 4 52 contain have 742:745 arg1 chains 735:740 PDB glycan chains 724:740 PDB glycan chains 724:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 4 52 contain have 742:745 arg2 derivative 773:782 at least one carbohydrate derivative 747:782 at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader 747:851 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 4 52 contain have 742:745 arg1 % 719:719 more than 50% 707:719 more than 50% of PDB glycan chains 707:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 4 53 theme Glycan 839:844 arg1 Reader 846:851 the original Glycan Reader 826:851 the original Glycan Reader 826:851 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 6 54 theme Glycan 1063:1068 arg1 Reader 1070:1075 CHARMM-GUI Glycan Reader 1052:1075 CHARMM-GUI Glycan Reader 1052:1075 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 11 55 theme @ 1868:1868 arg1 lehigh.edu 1869:1878 CONTACT wonpil@lehigh.edu 1854:1878 CONTACT wonpil@lehigh.edu. 1854:1879 CONTACT wonpil@lehigh.edu. 28582506 2 56 theme Data 262:265 arg1 Bank 267:270 Protein Data Bank 254:270 Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 254:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 56 theme Data 262:265 arg1 PDB 273:275 PDB 273:275 PDB 273:275 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 1 57 theme essential 156:164 arg1 processes 177:185 many essential biological processes 151:185 many essential biological processes 151:185 MOTIVATION Glycans play a central role in many essential biological processes. 28582506 9 58 theme glycan 1641:1646 arg1 tool 1664:1667 an intuitive glycan sequence search tool 1628:1667 an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB 1628:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 6 59 theme various 1135:1141 arg1 glycoconjugates 1143:1157 various glycoconjugates 1135:1157 various glycoconjugates with most sugar types and chemical modifications 1135:1206 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 5 60 theme RESULTS 854:860 arg1 Reader 869:874 RESULTS Glycan Reader 854:874 RESULTS Glycan Reader 854:874 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 6 61 with glycoconjugates 1143:1157 arg1 types 1175:1179 most sugar types 1164:1179 most sugar types 1164:1179 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 6 61 with glycoconjugates 1143:1157 arg1 modifications 1194:1206 chemical modifications 1185:1206 chemical modifications 1185:1206 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 4 62 theme original 830:837 arg1 Reader 846:851 the original Glycan Reader 826:851 the original Glycan Reader 826:851 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 7 63 theme anomeric 1399:1406 arg1 states 1408:1413 anomeric states 1399:1413 anomeric states 1399:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 4 64 theme PDB 626:628 arg1 files 630:634 current PDB files 618:634 current PDB files 618:634 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 9 65 theme search 1657:1662 arg1 tool 1664:1667 an intuitive glycan sequence search tool 1628:1667 an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB 1628:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 5 66 theme Glycan 862:867 arg1 Reader 869:874 RESULTS Glycan Reader 854:874 RESULTS Glycan Reader 854:874 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 9 67 theme glycan 1681:1686 arg1 structures 1688:1697 complex glycan structures 1673:1697 complex glycan structures with various chemical modifications in the PDB 1673:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 3 68 theme chemical 574:581 arg1 space 583:587 their chemical space 568:587 their chemical space much diverse 568:600 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 28582506 7 69 theme glycosidic 1374:1383 arg1 linkages 1385:1392 glycosidic linkages 1374:1392 glycosidic linkages 1374:1392 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 1 70 theme MOTIVATION 109:118 arg1 Glycans 120:126 MOTIVATION Glycans 109:126 MOTIVATION Glycans 109:126 MOTIVATION Glycans play a central role in many essential biological processes. 28582506 9 71 theme chemical 1712:1719 arg1 modifications 1721:1733 various chemical modifications 1704:1733 various chemical modifications in the PDB 1704:1744 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 7 72 theme modifications 1359:1371 arg1 addition/deletion/modification 1282:1311 addition/deletion/modification 1282:1311 addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1282:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 12 73 from online 1958:1963 arg1 available 1930:1938 available 1930:1938 available 1930:1938 SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 28582506 6 74 theme sugar 1169:1173 arg1 types 1175:1179 most sugar types 1164:1179 most sugar types 1164:1179 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 4 75 theme most 673:676 arg1 derivatives 691:701 most carbohydrate derivatives 673:701 most carbohydrate derivatives 673:701 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 1 76 theme central 135:141 arg1 role 143:146 a central role 133:146 a central role 133:146 MOTIVATION Glycans play a central role in many essential biological processes. 28582506 2 77 dep detection 325:333 arg1 the 311:313 the 311:313 the 311:313 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 0 78 theme Data 98:101 arg1 Bank 103:106 the Protein Data Bank 86:106 the Protein Data Bank 86:106 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 6 79 theme CHARMM-GUI 1052:1061 arg1 Reader 1070:1075 CHARMM-GUI Glycan Reader 1052:1075 CHARMM-GUI Glycan Reader 1052:1075 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 2 80 theme glycosidic 364:373 arg1 linkages 375:382 glycosidic linkages 364:382 glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 364:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 5 81 theme sugar 918:922 arg1 types 924:928 most sugar types 913:928 most sugar types 913:928 RESULTS Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. 28582506 7 82 theme glycosylation 1316:1328 arg1 types 1330:1334 glycosylation types 1316:1334 glycosylation types 1316:1334 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 2 83 theme sugars 353:358 arg1 annotation 339:348 annotation 339:348 annotation 339:348 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 83 theme sugars 353:358 arg1 detection 325:333 automatic detection 315:333 automatic detection 315:333 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 12 84 theme SUPPLEMENTARY 1881:1893 arg1 data 1921:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 28582506 9 85 from Database 1569:1576 arg1 GlycanStructure.Org 1581:1599 GlycanStructure.Org 1581:1599 GlycanStructure.Org 1581:1599 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 4 86 theme glycan 728:733 arg1 chains 735:740 PDB glycan chains 724:740 PDB glycan chains 724:740 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 12 87 theme Supplementary 1907:1919 arg1 data 1921:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data 1881:1924 SUPPLEMENTARY INFORMATION Supplementary data are available at Bioinformatics online. 28582506 2 88 theme atomic 434:439 arg1 coordinates 441:451 atomic coordinates 434:451 atomic coordinates 434:451 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 0 89 theme sugar 44:48 arg1 types 50:54 most sugar types 39:54 most sugar types 39:54 Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank. 28582506 9 90 theme Fragment 1560:1567 arg1 Database 1569:1576 Glycan Fragment Database 1553:1576 Glycan Fragment Database in GlycanStructure.Org 1553:1599 Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. 28582506 6 91 theme simulation 1104:1113 arg1 system 1115:1120 the simulation system 1100:1120 the simulation system 1100:1120 CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. 28582506 2 92 contain containing 284:293 arg2 glycans 295:301 glycans 295:301 glycans 295:301 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 2 92 contain containing 284:293 arg1 files 278:282 Protein Data Bank (PDB) files 254:282 Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information 254:480 Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. 28582506 4 93 theme carbohydrate 760:771 arg1 derivative 773:782 at least one carbohydrate derivative 747:782 at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader 747:851 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 11 94 theme wonpil 1862:1867 arg1 lehigh.edu 1869:1878 CONTACT wonpil@lehigh.edu 1854:1878 CONTACT wonpil@lehigh.edu. 1854:1879 CONTACT wonpil@lehigh.edu. 28582506 7 95 theme new 1226:1228 arg1 functionality 1230:1242 a new functionality 1224:1242 a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states 1224:1413 It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. 28582506 4 96 theme current 618:624 arg1 files 630:634 current PDB files 618:634 current PDB files 618:634 Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. 28582506 3 97 theme various 506:512 arg1 modifications 523:535 various chemical modifications 506:535 various chemical modifications 506:535 Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. 26959726 4 0 theme nonpolar 479:486 arg1 area 496:499 the nonpolar surface area 475:499 the nonpolar surface area of glycans 475:510 Also, the nonpolar surface area of glycans was calculated and compared to their experimental retention times. 26959726 8 1 theme N-glycan 931:938 arg1 compositions 940:951 78 N-glycan compositions 928:951 78 N-glycan compositions identified 928:962 In total, there were 78 N-glycan compositions identified. 26959726 1 2 theme N-glycan 139:146 arg1 elucidation 158:168 N-glycan structure elucidation 139:168 N-glycan structure elucidation 139:168 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 9 3 theme R 1100:1100 arg1 value 1105:1109 R(2) value 1100:1109 R(2) value for complex N-glycans 1100:1131 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 0 4 theme serum 71:75 arg1 glycoproteins 77:89 human blood serum glycoproteins 59:89 human blood serum glycoproteins 59:89 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 5 5 theme retention 584:592 arg1 time 594:597 retention time 584:597 retention time 584:597 Both retention time and nonpolar surface area trends are similar when the intermolecular interaction is included in the calculation. 26959726 1 6 theme structure 148:156 arg1 elucidation 158:168 N-glycan structure elucidation 139:168 N-glycan structure elucidation 139:168 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 4 7 theme retention 562:570 arg1 times 572:576 their experimental retention times 543:576 their experimental retention times 543:576 Also, the nonpolar surface area of glycans was calculated and compared to their experimental retention times. 26959726 1 8 theme low 244:246 arg1 abundance 248:256 low abundance 244:256 low abundance 244:256 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 11 9 used used 1302:1305 arg2 data 1292:1295 MS/MS data 1286:1295 MS/MS data 1286:1295 MS/MS data were used to confirm the structural isomers. 26959726 5 10 theme intermolecular 653:666 arg1 interaction 668:678 the intermolecular interaction 649:678 the intermolecular interaction 649:678 Both retention time and nonpolar surface area trends are similar when the intermolecular interaction is included in the calculation. 26959726 7 11 theme retention 851:859 arg1 time 861:864 retention time 851:864 retention time 851:864 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 1 12 theme powerful 120:127 arg1 tools 129:133 the most powerful tools 111:133 the most powerful tools for N-glycan structure elucidation 111:168 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 5 13 theme surface 612:618 arg1 area 620:623 nonpolar surface area 603:623 nonpolar surface area 603:623 Both retention time and nonpolar surface area trends are similar when the intermolecular interaction is included in the calculation. 26959726 5 14 theme nonpolar 603:610 arg1 area 620:623 nonpolar surface area 603:623 nonpolar surface area 603:623 Both retention time and nonpolar surface area trends are similar when the intermolecular interaction is included in the calculation. 26959726 9 15 theme glycan 1068:1073 arg1 structures 1075:1084 glycan structures 1068:1084 glycan structures 1068:1084 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 0 16 gly glycoproteins 77:89 arg1 glycoproteins 77:89 human blood serum glycoproteins 59:89 human blood serum glycoproteins 59:89 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 9 17 theme structures 1075:1084 arg1 subgroup 1056:1063 each subgroup 1051:1063 each subgroup of glycan structures 1051:1084 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 9 18 theme linear 967:972 arg1 relationship 974:985 A linear relationship 965:985 A linear relationship between retention times and molecular weights 965:1031 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 11 19 theme MS/MS 1286:1290 arg1 data 1292:1295 MS/MS data 1286:1295 MS/MS data 1286:1295 MS/MS data were used to confirm the structural isomers. 26959726 11 20 theme structural 1322:1331 arg1 isomers 1333:1339 the structural isomers 1318:1339 the structural isomers 1318:1339 MS/MS data were used to confirm the structural isomers. 26959726 7 21 theme high 828:831 arg1 accuracy 838:845 high mass accuracy 828:845 high mass accuracy 828:845 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 3 22 theme dextrin 424:430 arg1 times 382:386 retention times 372:386 retention times 372:386 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 0 23 theme N-glycans 26:34 arg1 LC-MS/MS 0:7 LC-MS/MS 0:7 LC-MS/MS of permethylated N-glycans 0:34 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 10 24 theme structural 1239:1248 arg1 isomers 1250:1256 structural isomers 1239:1256 structural isomers as well as linkage isomers 1239:1283 Moreover, the retention time could be further applied to distinguish between structural isomers as well as linkage isomers. 26959726 6 25 theme branch 769:774 arg1 types 776:780 branch types 769:780 branch types 769:780 Moreover, retention time corresponds to glycan types and branch types. 26959726 7 26 theme N-glycans 787:795 arg1 model 806:810 The N-glycans analysis model 783:810 The N-glycans analysis model 783:810 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 7 27 theme mass 833:836 arg1 accuracy 838:845 high mass accuracy 828:845 high mass accuracy 828:845 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 3 28 theme retention 372:380 arg1 times 382:386 retention times 372:386 retention times 372:386 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 4 29 theme experimental 549:560 arg1 times 572:576 their experimental retention times 543:576 their experimental retention times 543:576 Also, the nonpolar surface area of glycans was calculated and compared to their experimental retention times. 26959726 6 30 theme glycan 752:757 arg1 types 759:763 glycan types 752:763 glycan types 752:763 Moreover, retention time corresponds to glycan types and branch types. 26959726 2 31 theme N-glycans 336:344 arg1 behavior 310:317 the chromatographic behavior 290:317 the chromatographic behavior of permethylated N-glycans 290:344 In this study, we investigated the chromatographic behavior of permethylated N-glycans. 26959726 10 32 theme linkage 1269:1275 arg1 isomers 1277:1283 linkage isomers 1269:1283 structural isomers as well as linkage isomers 1239:1283 Moreover, the retention time could be further applied to distinguish between structural isomers as well as linkage isomers. 26959726 7 33 theme analysis 797:804 arg1 model 806:810 The N-glycans analysis model 783:810 The N-glycans analysis model 783:810 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 10 34 theme retention 1176:1184 arg1 time 1186:1189 the retention time 1172:1189 the retention time 1172:1189 Moreover, the retention time could be further applied to distinguish between structural isomers as well as linkage isomers. 26959726 7 35 theme serum 890:894 arg1 N-glycans 896:904 serum N-glycans 890:904 serum N-glycans 890:904 The N-glycans analysis model, which combines high mass accuracy and retention time, was applied to confirm serum N-glycans. 26959726 3 36 theme model 437:441 arg1 glycans 443:449 model glycans 437:449 model glycans 437:449 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 3 37 theme dextran 415:421 arg1 times 382:386 retention times 372:386 retention times 372:386 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 3 38 theme glycans 443:449 arg1 times 382:386 retention times 372:386 retention times 372:386 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 3 39 theme molecular 395:403 arg1 weight 405:410 molecular weight 395:410 molecular weight 395:410 The relationship between retention times versus molecular weight of dextran, dextrin, and model glycans was investigated. 26959726 4 40 theme glycans 504:510 arg1 area 496:499 the nonpolar surface area 475:499 the nonpolar surface area of glycans 475:510 Also, the nonpolar surface area of glycans was calculated and compared to their experimental retention times. 26959726 2 41 theme permethylated 322:334 arg1 N-glycans 336:344 permethylated N-glycans 322:344 permethylated N-glycans 322:344 In this study, we investigated the chromatographic behavior of permethylated N-glycans. 26959726 1 42 with structures 228:237 arg1 abundance 248:256 low abundance 244:256 low abundance 244:256 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 9 43 theme molecular 1015:1023 arg1 weights 1025:1031 molecular weights 1015:1031 molecular weights 1015:1031 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 2 44 theme chromatographic 294:308 arg1 behavior 310:317 the chromatographic behavior 290:317 the chromatographic behavior of permethylated N-glycans 290:344 In this study, we investigated the chromatographic behavior of permethylated N-glycans. 26959726 9 45 theme > 1154:1154 arg1 0.98 1156:1159 > 0.98 1154:1159 > 0.98 1154:1159 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 0 46 theme blood 65:69 arg1 glycoproteins 77:89 human blood serum glycoproteins 59:89 human blood serum glycoproteins 59:89 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 9 47 theme complex 1115:1121 arg1 N-glycans 1123:1131 complex N-glycans 1115:1131 complex N-glycans 1115:1131 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 1 48 theme tools 129:133 arg1 tools 129:133 the most powerful tools 111:133 the most powerful tools for N-glycan structure elucidation 111:168 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 1 48 theme tools 129:133 arg1 one 104:106 one 104:106 one 104:106 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 26959726 5 49 dep time 594:597 arg1 trends 625:630 trends 625:630 trends 625:630 Both retention time and nonpolar surface area trends are similar when the intermolecular interaction is included in the calculation. 26959726 9 50 theme retention 995:1003 arg1 times 1005:1009 retention times 995:1009 retention times 995:1009 A linear relationship between retention times and molecular weights were observed for each subgroup of glycan structures, for example, R(2) value for complex N-glycans was determined to be > 0.98. 26959726 0 51 theme human 59:63 arg1 glycoproteins 77:89 human blood serum glycoproteins 59:89 human blood serum glycoproteins 59:89 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 4 52 theme surface 488:494 arg1 area 496:499 the nonpolar surface area 475:499 the nonpolar surface area of glycans 475:510 Also, the nonpolar surface area of glycans was calculated and compared to their experimental retention times. 26959726 0 53 theme permethylated 12:24 arg1 N-glycans 26:34 permethylated N-glycans 12:34 permethylated N-glycans 12:34 LC-MS/MS of permethylated N-glycans derived from model and human blood serum glycoproteins. 26959726 6 54 theme retention 722:730 arg1 time 732:735 retention time 722:735 retention time 722:735 Moreover, retention time corresponds to glycan types and branch types. 26959726 1 55 theme glycan 221:226 arg1 structures 228:237 some glycan structures 216:237 some glycan structures with low abundance 216:256 LC-MS/MS is one of the most powerful tools for N-glycan structure elucidation; however, it is still challenging to identify some glycan structures with low abundance. 29043954 10 0 from changes 1607:1613 arg1 content 1641:1647 the chitosan cell-wall content 1618:1647 the chitosan cell-wall content 1618:1647 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 10 0 from changes 1607:1613 arg1 thickness 1660:1668 cell-wall thickness 1650:1668 cell-wall thickness 1650:1668 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 10 0 from changes 1607:1613 arg1 size 1682:1685 capsule size 1674:1685 capsule size 1674:1685 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 5 1 theme cellular 849:856 arg1 signalling 858:867 cellular signalling 849:867 cellular signalling in fungal and plant cells 849:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 4 2 theme sugar 609:613 arg1 N-acetylglucosamine 615:633 The amino sugar N-acetylglucosamine 599:633 The amino sugar N-acetylglucosamine (GlcNAc) 599:642 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 2 theme sugar 609:613 arg1 constituent 653:663 a key constituent 647:663 a key constituent of the cell-wall chitin 647:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 2 theme sugar 609:613 arg1 GlcNAc 636:641 GlcNAc 636:641 GlcNAc 636:641 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 9 3 theme carbon 1436:1441 arg1 source 1426:1431 a sole source 1419:1431 a sole source of carbon and nitrogen 1419:1454 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 9 3 theme carbon 1436:1441 arg1 GlcNAc 1409:1414 GlcNAc 1409:1414 GlcNAc 1409:1414 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 12 4 theme melanin 2059:2065 arg1 deposition 2067:2076 increased melanin deposition 2049:2076 increased melanin deposition 2049:2076 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 5 5 dep activator 823:831 arg1 an 820:821 an 820:821 an 820:821 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 9 6 from biospherenutritionally 1534:1555 arg1 component 1488:1496 a component 1486:1496 a component of a highly abundant polymer in the biospherenutritionally 1486:1555 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 2 7 theme second 320:325 arg1 factor 352:357 its second most important virulence factor 316:357 its second most important virulence factor 316:357 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 9 8 theme nitrogen 1447:1454 arg1 source 1426:1431 a sole source 1419:1431 a sole source of carbon and nitrogen 1419:1454 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 9 8 theme nitrogen 1447:1454 arg1 GlcNAc 1409:1414 GlcNAc 1409:1414 GlcNAc 1409:1414 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 4 9 theme chitin 682:687 arg1 N-acetylglucosamine 615:633 The amino sugar N-acetylglucosamine 599:633 The amino sugar N-acetylglucosamine (GlcNAc) 599:642 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 9 theme chitin 682:687 arg1 constituent 653:663 a key constituent 647:663 a key constituent of the cell-wall chitin 647:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 1 10 theme major 202:206 arg1 pathogen 208:215 a major pathogen 200:215 a major pathogen in immunocompromised patients 200:245 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 4 11 link N-linked 710:717 arg1 glycosylation 719:731 N-linked glycosylation 710:731 N-linked glycosylation 710:731 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 7 12 theme molecular 1086:1094 arg1 structure 1096:1104 its molecular structure 1082:1104 its molecular structure 1082:1104 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 3 13 theme stress 591:596 arg1 conditions 577:586 conditions 577:586 conditions of stress 577:596 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 5 14 theme signalling 858:867 arg1 mediator 837:844 mediator 837:844 mediator 837:844 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 5 14 theme signalling 858:867 arg1 activator 823:831 activator 823:831 activator 823:831 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 6 15 theme cell 983:986 arg1 wall 988:991 the cell wall 979:991 the cell wall 979:991 Furthermore, chitin and chitosan polysaccharides interact with melanin pigments in the cell wall and have been found to be essential for melanization. 29043954 5 16 theme fungal 872:877 arg1 cells 889:893 fungal and plant cells 872:893 fungal and plant cells 872:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 12 17 theme cell-wall 2016:2024 arg1 scaffold 2026:2033 the fungal cell-wall scaffold 2005:2033 the fungal cell-wall scaffold 2005:2033 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 8 18 theme supplementation 1300:1314 arg1 effect 1283:1288 the effect 1279:1288 the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization 1279:1369 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 10 19 theme cell-wall 1631:1639 arg1 content 1641:1647 the chitosan cell-wall content 1618:1647 the chitosan cell-wall content 1618:1647 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 11 20 theme cell 1865:1868 arg1 wall 1870:1873 the cell wall 1861:1873 the cell wall 1861:1873 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 6 21 theme melanin 959:965 arg1 pigments 967:974 melanin pigments 959:974 melanin pigments in the cell wall 959:991 Furthermore, chitin and chitosan polysaccharides interact with melanin pigments in the cell wall and have been found to be essential for melanization. 29043954 8 22 theme cryptococcal 1319:1330 arg1 composition 1342:1352 cryptococcal cell-wall composition 1319:1352 cryptococcal cell-wall composition 1319:1352 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 2 23 theme melanin 288:294 arg1 pigments 296:303 melanin pigments 288:303 melanin pigments 288:303 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 5 24 from cells 889:893 arg1 mediator 837:844 mediator 837:844 mediator 837:844 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 5 24 from cells 889:893 arg1 activator 823:831 activator 823:831 activator 823:831 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 9 25 theme abundant 1510:1517 arg1 polymer 1519:1525 a highly abundant polymer 1501:1525 a highly abundant polymer in the biospherenutritionally 1501:1555 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 5 26 theme suggested 779:787 arg1 roles 800:804 suggested additional roles 779:804 suggested additional roles for GlcNAc 779:815 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 13 27 theme pleiotropic 2131:2141 arg1 effects 2143:2149 pleiotropic effects 2131:2149 pleiotropic effects 2131:2149 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 7 28 theme nuclear 1191:1197 arg1 NMR 1219:1221 NMR 1219:1221 NMR 1219:1221 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 7 28 theme nuclear 1191:1197 arg1 resonance 1208:1216 advanced nuclear magnetic resonance 1182:1216 advanced nuclear magnetic resonance (NMR) 1182:1222 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 13 29 contain had 2127:2129 arg1 supplementation 2111:2125 GlcNAc supplementation 2104:2125 GlcNAc supplementation 2104:2125 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 13 29 contain had 2127:2129 arg2 effects 2143:2149 pleiotropic effects 2131:2149 pleiotropic effects 2131:2149 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 13 30 theme tracking 2303:2310 arg1 studies 2312:2318 metabolic tracking studies 2293:2318 metabolic tracking studies 2293:2318 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 4 31 theme N-linked 710:717 arg1 glycosylation 719:731 N-linked glycosylation 710:731 N-linked glycosylation 710:731 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 5 32 contain have 774:777 arg2 roles 800:804 suggested additional roles 779:804 suggested additional roles for GlcNAc 779:815 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 5 32 contain have 774:777 arg1 studies 766:772 Recent studies 759:772 Recent studies 759:772 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 1 33 from pathogen 208:215 arg1 patients 238:245 immunocompromised patients 220:245 immunocompromised patients 220:245 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 5 34 from mediator 837:844 arg1 cells 889:893 fungal and plant cells 872:893 fungal and plant cells 872:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 3 35 theme fungal 469:474 arg1 structure 497:505 a complex structure 487:505 a complex structure that is essential for maintaining cell morphology and viability under conditions of stress 487:596 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 35 theme fungal 469:474 arg1 essential 515:523 essential 515:523 essential 515:523 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 35 theme fungal 469:474 arg1 wall 481:484 the fungal cell wall 465:484 the fungal cell wall 465:484 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 5 36 from activator 823:831 arg1 cells 889:893 fungal and plant cells 872:893 fungal and plant cells 872:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 12 37 theme cell 1960:1963 arg1 separation 1965:1974 the mother-daughter cell separation 1940:1974 the mother-daughter cell separation 1940:1974 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 4 38 used used 696:699 arg2 GlcNAc 636:641 GlcNAc 636:641 GlcNAc 636:641 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 38 used used 696:699 arg2 constituent 653:663 a key constituent 647:663 a key constituent of the cell-wall chitin 647:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 38 used used 696:699 arg2 N-acetylglucosamine 615:633 The amino sugar N-acetylglucosamine 599:633 The amino sugar N-acetylglucosamine (GlcNAc) 599:642 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 0 39 theme Cryptococcus 28:39 arg1 composition 62:72 cell-wall composition 52:72 cell-wall composition 52:72 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 39 theme Cryptococcus 28:39 arg1 architecture 86:97 melanin architecture 78:97 melanin architecture 78:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 39 theme Cryptococcus 28:39 arg1 neoformans 41:50 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 12 40 with growth 1977:1982 arg1 GlcNAc 1989:1994 GlcNAc 1989:1994 GlcNAc 1989:1994 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 4 41 dep glycosylation 719:731 arg1 synthesis 748:756 synthesis 748:756 synthesis 748:756 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 1 42 theme phylum 171:176 arg1 Basidiomycetes 178:191 the phylum Basidiomycetes 167:191 the phylum Basidiomycetes 167:191 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 0 43 theme cell-wall 52:60 arg1 composition 62:72 cell-wall composition 52:72 cell-wall composition 52:72 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 43 theme cell-wall 52:60 arg1 neoformans 41:50 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 6 44 from pigments 967:974 arg1 wall 988:991 the cell wall 979:991 the cell wall 979:991 Furthermore, chitin and chitosan polysaccharides interact with melanin pigments in the cell wall and have been found to be essential for melanization. 29043954 11 45 theme 15N-labelled 1729:1740 arg1 GlcNAc 1742:1747 isotopically 15N-labelled GlcNAc 1716:1747 isotopically 15N-labelled GlcNAc 1716:1747 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 7 46 theme critical 1158:1165 arg1 insights 1167:1174 critical insights 1158:1174 critical insights 1158:1174 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 12 47 theme altered 1880:1886 arg1 ratio 1907:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 3 48 theme cell 541:544 arg1 morphology 546:555 cell morphology 541:555 cell morphology 541:555 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 5 49 theme plant 883:887 arg1 cells 889:893 fungal and plant cells 872:893 fungal and plant cells 872:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 9 50 theme sole 1421:1424 arg1 source 1426:1431 a sole source 1419:1431 a sole source of carbon and nitrogen 1419:1454 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 9 50 theme sole 1421:1424 arg1 GlcNAc 1409:1414 GlcNAc 1409:1414 GlcNAc 1409:1414 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 6 51 dep chitin 909:914 arg1 polysaccharides 929:943 polysaccharides 929:943 polysaccharides 929:943 Furthermore, chitin and chitosan polysaccharides interact with melanin pigments in the cell wall and have been found to be essential for melanization. 29043954 7 52 theme imaging 1228:1234 arg1 techniques 1236:1245 imaging techniques 1228:1245 imaging techniques 1228:1245 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 2 53 theme polysaccharide 384:397 arg1 capsule 399:405 a polysaccharide capsule 382:405 a polysaccharide capsule 382:405 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 4 54 theme amino 603:607 arg1 N-acetylglucosamine 615:633 The amino sugar N-acetylglucosamine 599:633 The amino sugar N-acetylglucosamine (GlcNAc) 599:642 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 54 theme amino 603:607 arg1 constituent 653:663 a key constituent 647:663 a key constituent of the cell-wall chitin 647:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 54 theme amino 603:607 arg1 GlcNAc 636:641 GlcNAc 636:641 GlcNAc 636:641 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 12 55 theme increased 2049:2057 arg1 deposition 2067:2076 increased melanin deposition 2049:2076 increased melanin deposition 2049:2076 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 9 56 from polymer 1519:1525 arg1 biospherenutritionally 1534:1555 the biospherenutritionally 1530:1555 the biospherenutritionally 1530:1555 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 4 57 theme cell-wall 672:680 arg1 chitin 682:687 the cell-wall chitin 668:687 the cell-wall chitin 668:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 10 58 theme chitosan 1622:1629 arg1 content 1641:1647 the chitosan cell-wall content 1618:1647 the chitosan cell-wall content 1618:1647 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 2 59 theme virulence 342:350 arg1 factor 352:357 its second most important virulence factor 316:357 its second most important virulence factor 316:357 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 1 60 theme immunocompromised 220:236 arg1 patients 238:245 immunocompromised patients 220:245 immunocompromised patients 220:245 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 4 61 theme key 649:651 arg1 N-acetylglucosamine 615:633 The amino sugar N-acetylglucosamine 599:633 The amino sugar N-acetylglucosamine (GlcNAc) 599:642 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 4 61 theme key 649:651 arg1 constituent 653:663 a key constituent 647:663 a key constituent of the cell-wall chitin 647:687 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 10 62 theme cell-wall 1650:1658 arg1 thickness 1660:1668 cell-wall thickness 1650:1668 cell-wall thickness 1650:1668 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 7 63 theme melanin 1073:1079 arg1 importance 1059:1068 the importance 1055:1068 the importance of melanin 1055:1079 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 2 64 theme neoformans 266:275 arg1 ability 252:258 The ability 248:258 The ability of C. neoformans to produce melanin pigments 248:303 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 12 65 theme fungal 2009:2014 arg1 scaffold 2026:2033 the fungal cell-wall scaffold 2005:2033 the fungal cell-wall scaffold 2005:2033 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 8 66 theme GlcNAc 1293:1298 arg1 supplementation 1300:1314 GlcNAc supplementation 1293:1314 GlcNAc supplementation 1293:1314 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 13 67 theme melanin 2168:2174 arg1 architectures 2176:2188 cell-wall and melanin architectures 2154:2188 cell-wall and melanin architectures 2154:2188 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 5 68 from signalling 858:867 arg1 cells 889:893 fungal and plant cells 872:893 fungal and plant cells 872:893 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 4 69 theme GPI 737:739 arg1 anchor 741:746 GPI anchor 737:746 GPI anchor 737:746 The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. 29043954 11 70 theme building 1801:1808 arg1 block 1810:1814 a building block 1799:1814 a building block for chitin/chitosan 1799:1834 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 11 70 theme building 1801:1808 arg1 monomer 1781:1787 the exogenous monomer 1767:1787 the exogenous monomer 1767:1787 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 8 71 from effect 1283:1288 arg1 composition 1342:1352 cryptococcal cell-wall composition 1319:1352 cryptococcal cell-wall composition 1319:1352 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 8 71 from effect 1283:1288 arg1 melanization 1358:1369 melanization 1358:1369 melanization 1358:1369 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 9 72 theme polymer 1519:1525 arg1 component 1488:1496 a component 1486:1496 a component of a highly abundant polymer in the biospherenutritionally 1486:1555 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 8 73 theme cell-wall 1332:1340 arg1 composition 1342:1352 cryptococcal cell-wall composition 1319:1352 cryptococcal cell-wall composition 1319:1352 In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. 29043954 5 74 theme additional 789:798 arg1 roles 800:804 suggested additional roles 779:804 suggested additional roles for GlcNAc 779:815 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 13 75 theme cell-wall 2154:2162 arg1 architectures 2176:2188 cell-wall and melanin architectures 2154:2188 cell-wall and melanin architectures 2154:2188 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 2 76 theme important 332:340 arg1 factor 352:357 its second most important virulence factor 316:357 its second most important virulence factor 316:357 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 29043954 5 77 theme Recent 759:764 arg1 studies 766:772 Recent studies 759:772 Recent studies 759:772 Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. 29043954 13 78 theme GlcNAc 2104:2109 arg1 supplementation 2111:2125 GlcNAc supplementation 2104:2125 GlcNAc supplementation 2104:2125 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 12 79 theme mother-daughter 1944:1958 arg1 separation 1965:1974 the mother-daughter cell separation 1940:1974 the mother-daughter cell separation 1940:1974 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 1 80 theme Cryptococcus 100:111 arg1 neoformans 113:122 Cryptococcus neoformans 100:122 Cryptococcus neoformans 100:122 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 1 80 theme Cryptococcus 100:111 arg1 fungus 144:149 an environmental fungus 127:149 an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients 127:245 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 10 81 theme capsule 1674:1680 arg1 size 1682:1685 capsule size 1674:1685 capsule size 1674:1685 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 7 82 theme magnetic 1199:1206 arg1 NMR 1219:1221 NMR 1219:1221 NMR 1219:1221 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 7 82 theme magnetic 1199:1206 arg1 resonance 1208:1216 advanced nuclear magnetic resonance 1182:1216 advanced nuclear magnetic resonance (NMR) 1182:1222 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 7 83 theme advanced 1182:1189 arg1 NMR 1219:1221 NMR 1219:1221 NMR 1219:1221 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 7 83 theme advanced 1182:1189 arg1 resonance 1208:1216 advanced nuclear magnetic resonance 1182:1216 advanced nuclear magnetic resonance (NMR) 1182:1222 Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. 29043954 13 84 theme metabolic 2293:2301 arg1 studies 2312:2318 metabolic tracking studies 2293:2318 metabolic tracking studies 2293:2318 In summary, GlcNAc supplementation had pleiotropic effects on cell-wall and melanin architectures, and thus established its capacity to perturb these structures, a property that could prove useful for metabolic tracking studies. 29043954 12 85 theme negative 1920:1927 arg1 effects 1929:1935 no negative effects 1917:1935 no negative effects 1917:1935 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 1 86 theme environmental 130:142 arg1 neoformans 113:122 Cryptococcus neoformans 100:122 Cryptococcus neoformans 100:122 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 1 86 theme environmental 130:142 arg1 fungus 144:149 an environmental fungus 127:149 an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients 127:245 Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. 29043954 3 87 theme complex 489:495 arg1 structure 497:505 a complex structure 487:505 a complex structure that is essential for maintaining cell morphology and viability under conditions of stress 487:596 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 87 theme complex 489:495 arg1 essential 515:523 essential 515:523 essential 515:523 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 87 theme complex 489:495 arg1 wall 481:484 the fungal cell wall 465:484 the fungal cell wall 465:484 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 0 88 theme melanin 78:84 arg1 architecture 86:97 melanin architecture 78:97 melanin architecture 78:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 88 theme melanin 78:84 arg1 neoformans 41:50 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 12 89 theme chitin-to-chitosan 1888:1905 arg1 ratio 1907:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 9 90 from component 1488:1496 arg1 biospherenutritionally 1534:1555 the biospherenutritionally 1530:1555 the biospherenutritionally 1530:1555 C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. 29043954 11 91 theme exogenous 1771:1779 arg1 block 1810:1814 a building block 1799:1814 a building block for chitin/chitosan 1799:1834 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 11 91 theme exogenous 1771:1779 arg1 monomer 1781:1787 the exogenous monomer 1767:1787 the exogenous monomer 1767:1787 Supplementing cultures with isotopically 15N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. 29043954 12 92 contain had 1913:1915 arg1 ratio 1907:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio 1876:1911 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 12 92 contain had 1913:1915 arg2 effects 1929:1935 no negative effects 1917:1935 no negative effects 1917:1935 The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. 29043954 0 93 dep neoformans 41:50 arg1 composition 62:72 cell-wall composition 52:72 cell-wall composition 52:72 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 93 dep neoformans 41:50 arg1 architecture 86:97 melanin architecture 78:97 melanin architecture 78:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 0 93 dep neoformans 41:50 arg1 neoformans 41:50 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 Cryptococcus neoformans cell-wall composition and melanin architecture 28:97 N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture. 29043954 3 94 theme cell 476:479 arg1 structure 497:505 a complex structure 487:505 a complex structure that is essential for maintaining cell morphology and viability under conditions of stress 487:596 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 94 theme cell 476:479 arg1 essential 515:523 essential 515:523 essential 515:523 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 3 94 theme cell 476:479 arg1 wall 481:484 the fungal cell wall 465:484 the fungal cell wall 465:484 Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. 29043954 10 95 theme neoformans 1561:1570 arg1 cells 1572:1576 C. neoformans cells 1558:1576 C. neoformans cells grown with GlcNAc 1558:1594 C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. 29043954 2 96 theme capsule 399:405 arg1 presence 370:377 the presence 366:377 the presence of a polysaccharide capsule 366:405 The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. 25355867 7 0 theme apoplast/plasma 1218:1232 arg1 membrane 1234:1241 the apoplast/plasma membrane 1214:1241 the apoplast/plasma membrane 1214:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 7 1 from membrane 1234:1241 arg1 located 1203:1209 located 1203:1209 located 1203:1209 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 7 2 contain carried 1110:1116 arg1 A1AT 1105:1108 Secreted A1AT 1096:1108 Secreted A1AT 1096:1108 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 7 2 contain carried 1110:1116 arg2 N-glycans 1148:1156 vacuolar-type paucimannosidic N-glycans 1118:1156 vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane 1118:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 9 3 theme plant 1455:1459 arg1 processes 1475:1483 plant glycosylation processes 1455:1483 plant glycosylation processes 1455:1483 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 2 4 from cases 364:368 arg1 difficult 346:354 difficult 346:354 difficult 346:354 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 5 5 theme center 814:819 arg1 loop 821:824 the reactive center loop 801:824 the reactive center loop 801:824 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 8 6 theme glycoengineering 1274:1289 arg1 approach 1291:1298 an intensive glycoengineering approach 1261:1298 an intensive glycoengineering approach 1261:1298 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 5 7 theme full-length 684:694 arg1 A1AT 696:699 full-length A1AT 684:699 two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity 673:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 2 8 theme factors 237:243 arg1 knowledge 210:218 our limited knowledge 198:218 our limited knowledge 198:218 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 9 9 theme postendoplasmic 1515:1529 arg1 processing 1549:1558 postendoplasmic reticulum protein processing 1515:1558 postendoplasmic reticulum protein processing in plants 1515:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 6 10 theme terminal 907:914 arg1 consequence 952:962 a consequence 950:962 a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway) 950:1093 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 6 10 theme terminal 907:914 arg1 processing 924:933 the terminal protein processing 903:933 the terminal protein processing 903:933 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 5 11 theme material 660:667 arg1 g 646:646 up to 6 mg g(-1) 635:650 up to 6 mg g(-1) of leaf material 635:667 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 9 12 theme protein 1541:1547 arg1 processing 1549:1558 postendoplasmic reticulum protein processing 1515:1558 postendoplasmic reticulum protein processing in plants 1515:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 8 13 link serum-derived 1384:1396 arg1 counterpart 1398:1408 its serum-derived counterpart 1380:1408 its serum-derived counterpart 1380:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 7 14 from located 1203:1209 arg1 membrane 1234:1241 the apoplast/plasma membrane 1214:1241 the apoplast/plasma membrane 1214:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 7 15 theme Secreted 1096:1103 arg1 A1AT 1105:1108 Secreted A1AT 1096:1108 Secreted A1AT 1096:1108 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 8 16 theme secreted 1307:1314 arg1 A1AT 1316:1319 secreted A1AT 1307:1319 secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart 1307:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 1 17 theme expression 137:146 arg1 Plants 96:101 Plants 96:101 Plants 96:101 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 1 17 theme expression 137:146 arg1 system 148:153 an expression system 134:153 an expression system for complex recombinant proteins 134:186 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 7 18 theme paucimannosidic 1132:1146 arg1 N-glycans 1148:1156 vacuolar-type paucimannosidic N-glycans 1118:1156 vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane 1118:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 6 19 theme intrinsic 971:979 arg1 function 981:988 the intrinsic function 967:988 the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway) 967:1093 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 8 20 theme N-glycan 1341:1348 arg1 structures 1350:1359 sialylated N-glycan structures 1330:1359 sialylated N-glycan structures largely resembling its serum-derived counterpart 1330:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 5 21 theme inhibitory 749:758 arg1 activity 760:767 inhibitory activity 749:767 inhibitory activity 749:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 3 22 from expression 404:413 arg1 benthamiana 482:492 Nicotiana benthamiana 472:492 Nicotiana benthamiana 472:492 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 1 23 theme recombinant 167:177 arg1 proteins 179:186 complex recombinant proteins 159:186 complex recombinant proteins 159:186 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 9 24 from plants 1563:1568 arg1 aspects 1504:1510 important aspects 1494:1510 important aspects of postendoplasmic reticulum protein processing in plants 1494:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 5 25 theme located 701:707 arg1 A1AT 696:699 full-length A1AT 684:699 two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity 673:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 6 26 dep interaction 1008:1018 arg1 i.e. 999:1002 i.e. 999:1002 i.e. 999:1002 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 3 27 theme protease 428:435 arg1 A1AT 463:466 A1AT 463:466 A1AT 463:466 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 3 27 theme protease 428:435 arg1 α1-antitrypsin 447:460 the human protease inhibitor α1-antitrypsin 418:460 the human protease inhibitor α1-antitrypsin (A1AT) 418:467 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 8 28 gly sialylated 1330:1339 arg1 structures 1350:1359 sialylated N-glycan structures 1330:1359 sialylated N-glycan structures largely resembling its serum-derived counterpart 1330:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 5 29 theme recombinant 592:602 arg1 A1AT 604:607 recombinant A1AT 592:607 recombinant A1AT 592:607 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 6 30 theme [most 1035:1039 arg1 proteases 1055:1063 [most likely serine proteases] 1035:1064 [most likely serine proteases] along the secretory pathway 1035:1092 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 0 31 theme Nicotiana 73:81 arg1 benthamiana 83:93 Nicotiana benthamiana 73:93 Nicotiana benthamiana 73:93 Proteolytic and N-glycan processing of human α1-antitrypsin expressed in Nicotiana benthamiana. 25355867 0 32 theme Proteolytic 0:10 arg1 processing 25:34 Proteolytic and N-glycan processing 0:34 processing 25:34 Proteolytic and N-glycan processing of human α1-antitrypsin expressed in Nicotiana benthamiana. 25355867 0 33 theme N-glycan 16:23 arg1 processing 25:34 Proteolytic and N-glycan processing 0:34 processing 25:34 Proteolytic and N-glycan processing of human α1-antitrypsin expressed in Nicotiana benthamiana. 25355867 3 34 theme α1-antitrypsin 447:460 arg1 expression 404:413 the recombinant expression 388:413 the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana 388:492 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 9 35 from insights 1443:1450 arg1 processes 1475:1483 plant glycosylation processes 1455:1483 plant glycosylation processes 1455:1483 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 4 36 theme intensive 524:532 arg1 modifications 552:564 intensive posttranslational modifications 524:564 intensive posttranslational modifications 524:564 This serum protein undergoes intensive posttranslational modifications. 25355867 5 37 theme secreted 774:781 arg1 A1AT 783:786 secreted A1AT 774:786 secreted A1AT 774:786 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 2 38 theme process 331:337 arg1 design 339:344 process design 331:344 process design 331:344 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 6 39 with interaction 1008:1018 arg1 proteases 1025:1033 proteases 1025:1033 proteases 1025:1033 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 5 40 dep 6 641:641 arg1 to 638:639 to 638:639 to 638:639 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 9 41 theme unique 1436:1441 arg1 insights 1443:1450 unique insights 1436:1450 unique insights in plant glycosylation processes 1436:1483 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 6 42 theme serine 1048:1053 arg1 proteases 1055:1063 [most likely serine proteases] 1035:1064 [most likely serine proteases] along the secretory pathway 1035:1092 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 2 43 from difficult 346:354 arg1 cases 364:368 some cases 359:368 some cases 359:368 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 9 44 theme glycosylation 1461:1473 arg1 processes 1475:1483 plant glycosylation processes 1455:1483 plant glycosylation processes 1455:1483 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 0 45 theme α1-antitrypsin 45:58 arg1 processing 25:34 Proteolytic and N-glycan processing 0:34 processing 25:34 Proteolytic and N-glycan processing of human α1-antitrypsin expressed in Nicotiana benthamiana. 25355867 9 46 from processing 1549:1558 arg1 plants 1563:1568 plants 1563:1568 plants 1563:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 7 47 theme located 1203:1209 arg1 hexosaminidases 1187:1201 hexosaminidases 1187:1201 hexosaminidases located in the apoplast/plasma membrane 1187:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 5 48 theme mg 643:644 arg1 g 646:646 up to 6 mg g(-1) 635:650 up to 6 mg g(-1) of leaf material 635:667 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 2 49 theme product 304:310 arg1 integrity 312:320 product integrity 304:320 product integrity 304:320 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 5 50 theme reactive 805:812 arg1 loop 821:824 the reactive center loop 801:824 the reactive center loop 801:824 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 5 51 theme target 869:874 arg1 proteinases 876:886 target proteinases 869:886 target proteinases 869:886 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 6 52 theme protein 916:922 arg1 consequence 952:962 a consequence 950:962 a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway) 950:1093 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 6 52 theme protein 916:922 arg1 processing 924:933 the terminal protein processing 903:933 the terminal protein processing 903:933 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 8 53 contain carrying 1321:1328 arg2 structures 1350:1359 sialylated N-glycan structures 1330:1359 sialylated N-glycan structures largely resembling its serum-derived counterpart 1330:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 8 53 contain carrying 1321:1328 arg1 A1AT 1316:1319 secreted A1AT 1307:1319 secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart 1307:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 8 54 theme intensive 1264:1272 arg1 approach 1291:1298 an intensive glycoengineering approach 1261:1298 an intensive glycoengineering approach 1261:1298 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 5 55 theme leaf 655:658 arg1 material 660:667 leaf material 655:667 leaf material 655:667 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 5 56 from located 701:707 arg1 reticulum 728:736 the endoplasmic reticulum 712:736 the endoplasmic reticulum displaying inhibitory activity 712:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 2 57 theme intrinsic 227:235 arg1 factors 237:243 the intrinsic factors 223:243 the intrinsic factors that act along the secretory pathway 223:280 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 5 58 theme high 577:580 arg1 levels 582:587 Unusually high levels 567:587 Unusually high levels of recombinant A1AT 567:607 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 9 59 theme reticulum 1531:1539 arg1 processing 1549:1558 postendoplasmic reticulum protein processing 1515:1558 postendoplasmic reticulum protein processing in plants 1515:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 2 60 theme secretory 264:272 arg1 pathway 274:280 the secretory pathway 260:280 the secretory pathway 260:280 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 9 61 theme processing 1549:1558 arg1 aspects 1504:1510 important aspects 1494:1510 important aspects of postendoplasmic reticulum protein processing in plants 1494:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 6 62 theme A1AT 993:996 arg1 function 981:988 the intrinsic function 967:988 the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway) 967:1093 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 5 63 theme endoplasmic 716:726 arg1 reticulum 728:736 the endoplasmic reticulum 712:736 the endoplasmic reticulum displaying inhibitory activity 712:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 5 64 from reticulum 728:736 arg1 located 701:707 located 701:707 located 701:707 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 9 65 theme important 1494:1502 arg1 aspects 1504:1510 important aspects 1494:1510 important aspects of postendoplasmic reticulum protein processing in plants 1494:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 6 66 theme function 981:988 arg1 consequence 952:962 a consequence 950:962 a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway) 950:1093 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 6 66 theme function 981:988 arg1 processing 924:933 the terminal protein processing 903:933 the terminal protein processing 903:933 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 4 67 theme serum 500:504 arg1 protein 506:512 This serum protein 495:512 This serum protein 495:512 This serum protein undergoes intensive posttranslational modifications. 25355867 7 68 theme hexosaminidases 1187:1201 arg1 activity 1175:1182 the activity 1171:1182 the activity of hexosaminidases located in the apoplast/plasma membrane 1171:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 1 69 theme complex 159:165 arg1 proteins 179:186 complex recombinant proteins 159:186 complex recombinant proteins 159:186 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 5 70 dep forms 677:681 arg1 A1AT 696:699 full-length A1AT 684:699 two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity 673:767 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 8 71 theme sialylated 1330:1339 arg1 structures 1350:1359 sialylated N-glycan structures 1330:1359 sialylated N-glycan structures largely resembling its serum-derived counterpart 1330:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 1 72 used used 126:129 arg2 Plants 96:101 Plants 96:101 Plants 96:101 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 1 72 used used 126:129 arg2 system 148:153 an expression system 134:153 an expression system for complex recombinant proteins 134:186 Plants are increasingly being used as an expression system for complex recombinant proteins. 25355867 6 73 theme likely 1041:1046 arg1 proteases 1055:1063 [most likely serine proteases] 1035:1064 [most likely serine proteases] along the secretory pathway 1035:1092 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 6 74 dep A1AT 993:996 arg1 proteases 1055:1063 [most likely serine proteases] 1035:1064 [most likely serine proteases] along the secretory pathway 1035:1092 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 3 75 theme inhibitor 437:445 arg1 A1AT 463:466 A1AT 463:466 A1AT 463:466 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 3 75 theme inhibitor 437:445 arg1 α1-antitrypsin 447:460 the human protease inhibitor α1-antitrypsin 418:460 the human protease inhibitor α1-antitrypsin (A1AT) 418:467 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 5 76 theme A1AT 604:607 arg1 levels 582:587 Unusually high levels 567:587 Unusually high levels of recombinant A1AT 567:607 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 25355867 8 77 theme serum-derived 1384:1396 arg1 counterpart 1398:1408 its serum-derived counterpart 1380:1408 its serum-derived counterpart 1380:1408 Notwithstanding, an intensive glycoengineering approach led to secreted A1AT carrying sialylated N-glycan structures largely resembling its serum-derived counterpart. 25355867 3 78 theme human 422:426 arg1 A1AT 463:466 A1AT 463:466 A1AT 463:466 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 3 78 theme human 422:426 arg1 α1-antitrypsin 447:460 the human protease inhibitor α1-antitrypsin 418:460 the human protease inhibitor α1-antitrypsin (A1AT) 418:467 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 7 79 theme vacuolar-type 1118:1130 arg1 N-glycans 1148:1156 vacuolar-type paucimannosidic N-glycans 1118:1156 vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane 1118:1241 Secreted A1AT carried vacuolar-type paucimannosidic N-glycans generated by the activity of hexosaminidases located in the apoplast/plasma membrane. 25355867 0 80 theme human 39:43 arg1 α1-antitrypsin 45:58 human α1-antitrypsin 39:58 human α1-antitrypsin 39:58 Proteolytic and N-glycan processing of human α1-antitrypsin expressed in Nicotiana benthamiana. 25355867 4 81 theme posttranslational 534:550 arg1 modifications 552:564 intensive posttranslational modifications 524:564 intensive posttranslational modifications 524:564 This serum protein undergoes intensive posttranslational modifications. 25355867 3 82 theme recombinant 392:402 arg1 expression 404:413 the recombinant expression 388:413 the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana 388:492 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 9 83 from aspects 1504:1510 arg1 plants 1563:1568 plants 1563:1568 plants 1563:1568 In summary, we elucidate unique insights in plant glycosylation processes and show important aspects of postendoplasmic reticulum protein processing in plants. 25355867 6 84 theme secretory 1076:1084 arg1 pathway 1086:1092 the secretory pathway 1072:1092 the secretory pathway 1072:1092 We found that the terminal protein processing is most likely a consequence of the intrinsic function of A1AT (i.e. its interaction with proteases [most likely serine proteases] along the secretory pathway). 25355867 2 85 theme limited 202:208 arg1 knowledge 210:218 our limited knowledge 198:218 our limited knowledge 198:218 However, our limited knowledge of the intrinsic factors that act along the secretory pathway, which may compromise product integrity, renders process design difficult in some cases. 25355867 3 86 theme Nicotiana 472:480 arg1 benthamiana 482:492 Nicotiana benthamiana 472:492 Nicotiana benthamiana 472:492 Here, we pursued the recombinant expression of the human protease inhibitor α1-antitrypsin (A1AT) in Nicotiana benthamiana. 25355867 5 87 dep leaves 627:632 arg1 g 646:646 up to 6 mg g(-1) 635:650 up to 6 mg g(-1) of leaf material 635:667 Unusually high levels of recombinant A1AT were expressed in leaves (up to 6 mg g(-1) of leaf material) in two forms: full-length A1AT located in the endoplasmic reticulum displaying inhibitory activity, and secreted A1AT processed in the reactive center loop, thus rendering it unable to interact with target proteinases. 24952773 1 0 theme high-pressure 178:190 arg1 treatment 210:218 previous dynamic high-pressure microfluidization treatment 161:218 previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) 161:244 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 1 theme subsequent 250:259 arg1 glycation 261:269 subsequent glycation 250:269 subsequent glycation 250:269 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 2 theme microfluidization 192:208 arg1 treatment 210:218 previous dynamic high-pressure microfluidization treatment 161:218 previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) 161:244 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 3 theme glycation 261:269 arg1 effect 151:156 The combined effect 138:156 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) 138:351 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 0 4 theme glycation 117:125 arg1 treatment 127:135 glycation treatment 117:135 glycation treatment 117:135 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 4 5 theme circular 727:734 arg1 analysis 746:753 circular dichroism analysis 727:753 circular dichroism analysis 727:753 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 6 theme fluorescence 676:687 arg1 spectra 689:695 fluorescence spectra 676:695 fluorescence spectra 676:695 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 5 7 theme antigenicity 881:892 arg1 alteration 867:876 the alteration 863:876 the alteration of antigenicity 863:892 The conformational changes may contribute to the alteration of antigenicity. 24952773 4 8 theme UV 708:709 arg1 absorption 711:720 decreased UV absorption 698:720 decreased UV absorption 698:720 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 1 9 theme treatment 210:218 arg1 effect 151:156 The combined effect 138:156 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) 138:351 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 10 from effect 151:156 arg1 antigenicity 314:325 the antigenicity 310:325 the antigenicity of β-lactoglobulin (β-LG) 310:351 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 4 11 theme β-LG 812:815 arg1 changes 801:807 tertiary and secondary structural changes 767:807 tertiary and secondary structural changes of β-LG 767:815 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 3 12 theme Surface 448:454 arg1 content 473:479 Surface sulfhydryl group content 448:479 Surface sulfhydryl group content of β-LG-GOS 448:491 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 4 13 theme β-LG-GOS 588:595 arg1 samples 597:603 β-LG-GOS samples 588:603 β-LG-GOS samples 588:603 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 2 14 theme β-LG-GOS 391:398 arg1 antigenicity 375:386 The antigenicity 371:386 The antigenicity of β-LG-GOS 371:398 The antigenicity of β-LG-GOS decreased at relatively low pressure (≤120MPa). 24952773 4 15 theme tertiary 767:774 arg1 changes 801:807 tertiary and secondary structural changes 767:807 tertiary and secondary structural changes of β-LG 767:815 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 16 theme decreased 698:706 arg1 absorption 711:720 decreased UV absorption 698:720 decreased UV absorption 698:720 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 17 theme structural 790:799 arg1 changes 801:807 tertiary and secondary structural changes 767:807 tertiary and secondary structural changes of β-LG 767:815 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 0 18 theme conformational 17:30 arg1 changes 32:38 conformational changes 17:38 conformational changes 17:38 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 4 19 theme secondary 780:788 arg1 changes 801:807 tertiary and secondary structural changes 767:807 tertiary and secondary structural changes of β-LG 767:815 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 3 20 theme β-LG-GOS 533:540 arg1 hydrophobicity 515:528 surface hydrophobicity 507:528 surface hydrophobicity of β-LG-GOS 507:540 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 0 21 theme β-lactoglobulin 43:57 arg1 Antigenicity 0:11 Antigenicity 0:11 Antigenicity 0:11 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 0 21 theme β-lactoglobulin 43:57 arg1 changes 32:38 conformational changes 17:38 conformational changes 17:38 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 3 22 theme surface 507:513 arg1 hydrophobicity 515:528 surface hydrophobicity 507:528 surface hydrophobicity of β-LG-GOS 507:540 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 3 23 theme sulfhydryl 456:465 arg1 content 473:479 Surface sulfhydryl group content 448:479 Surface sulfhydryl group content of β-LG-GOS 448:491 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 2 24 theme low 424:426 arg1 ≤120MPa 438:444 ≤120MPa 438:444 ≤120MPa 438:444 The antigenicity of β-LG-GOS decreased at relatively low pressure (≤120MPa). 24952773 2 24 theme low 424:426 arg1 pressure 428:435 relatively low pressure 413:435 relatively low pressure (≤120MPa) 413:445 The antigenicity of β-LG-GOS decreased at relatively low pressure (≤120MPa). 24952773 5 25 theme conformational 822:835 arg1 changes 837:843 The conformational changes 818:843 The conformational changes 818:843 The conformational changes may contribute to the alteration of antigenicity. 24952773 3 26 theme group 467:471 arg1 content 473:479 Surface sulfhydryl group content 448:479 Surface sulfhydryl group content of β-LG-GOS 448:491 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 0 27 theme dynamic 62:68 arg1 microfluidization 84:100 dynamic high pressure microfluidization 62:100 dynamic high pressure microfluidization combining with glycation treatment 62:135 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 1 28 dep treatment 210:218 arg1 40 221:222 40 221:222 40 221:222 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 28 dep treatment 210:218 arg1 120 229:231 120 229:231 120 229:231 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 28 dep treatment 210:218 arg1 160MPa 238:243 160MPa 238:243 160MPa 238:243 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 28 dep treatment 210:218 arg1 80 225:226 80 225:226 80 225:226 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 4 29 theme protein 567:573 arg1 unfolding 575:583 protein unfolding 567:583 protein unfolding in β-LG-GOS samples 567:603 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 1 30 theme combined 142:149 arg1 effect 151:156 The combined effect 138:156 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) 138:351 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 31 theme β-lactoglobulin 330:344 arg1 antigenicity 314:325 the antigenicity 310:325 the antigenicity of β-lactoglobulin (β-LG) 310:351 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 32 with treatment 210:218 arg1 GOS 302:304 GOS 302:304 GOS 302:304 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 32 with treatment 210:218 arg1 galacto-oligosaccharides 276:299 galacto-oligosaccharides 276:299 galacto-oligosaccharides (GOS) 276:305 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 33 with glycation 261:269 arg1 GOS 302:304 GOS 302:304 GOS 302:304 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 1 33 with glycation 261:269 arg1 galacto-oligosaccharides 276:299 galacto-oligosaccharides 276:299 galacto-oligosaccharides (GOS) 276:305 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 3 34 theme β-LG-GOS 484:491 arg1 content 473:479 Surface sulfhydryl group content 448:479 Surface sulfhydryl group content of β-LG-GOS 448:491 Surface sulfhydryl group content of β-LG-GOS increased and surface hydrophobicity of β-LG-GOS decreased. 24952773 4 35 theme fluorescence 635:646 arg1 intensity 648:656 fluorescence intensity 635:656 fluorescence intensity 635:656 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 36 from unfolding 575:583 arg1 samples 597:603 β-LG-GOS samples 588:603 β-LG-GOS samples 588:603 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 0 37 theme pressure 75:82 arg1 microfluidization 84:100 dynamic high pressure microfluidization 62:100 dynamic high pressure microfluidization combining with glycation treatment 62:135 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 1 38 theme previous 161:168 arg1 treatment 210:218 previous dynamic high-pressure microfluidization treatment 161:218 previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) 161:244 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 4 39 theme spectra 689:695 arg1 analysis 746:753 circular dichroism analysis 727:753 circular dichroism analysis 727:753 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 39 theme spectra 689:695 arg1 absorption 711:720 decreased UV absorption 698:720 decreased UV absorption 698:720 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 39 theme spectra 689:695 arg1 quenching 622:630 quenching 622:630 quenching of fluorescence intensity 622:656 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 39 theme spectra 689:695 arg1 red-shift 663:671 the red-shift 659:671 the red-shift of fluorescence spectra 659:695 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 0 40 theme high 70:73 arg1 microfluidization 84:100 dynamic high pressure microfluidization 62:100 dynamic high pressure microfluidization combining with glycation treatment 62:135 Antigenicity and conformational changes of β-lactoglobulin by dynamic high pressure microfluidization combining with glycation treatment. 24952773 4 41 theme intensity 648:656 arg1 analysis 746:753 circular dichroism analysis 727:753 circular dichroism analysis 727:753 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 41 theme intensity 648:656 arg1 absorption 711:720 decreased UV absorption 698:720 decreased UV absorption 698:720 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 41 theme intensity 648:656 arg1 quenching 622:630 quenching 622:630 quenching of fluorescence intensity 622:656 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 4 41 theme intensity 648:656 arg1 red-shift 663:671 the red-shift 659:671 the red-shift of fluorescence spectra 659:695 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 24952773 1 42 theme dynamic 170:176 arg1 treatment 210:218 previous dynamic high-pressure microfluidization treatment 161:218 previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) 161:244 The combined effect of previous dynamic high-pressure microfluidization treatment (40, 80, 120, and 160MPa) and subsequent glycation with galacto-oligosaccharides (GOS) on the antigenicity of β-lactoglobulin (β-LG) was investigated. 24952773 4 43 theme dichroism 736:744 arg1 analysis 746:753 circular dichroism analysis 727:753 circular dichroism analysis 727:753 Additionally, protein unfolding in β-LG-GOS samples was reflected by quenching of fluorescence intensity, the red-shift of fluorescence spectra, decreased UV absorption, and circular dichroism analysis, indicating tertiary and secondary structural changes of β-LG. 28548838 0 0 theme Cell 72:75 arg1 Polysaccharide 85:98 A Cell Surface Polysaccharide 70:98 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 0:126 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 3 1 theme synthetic 655:663 arg1 targets 665:671 Both synthetic targets 650:671 Both synthetic targets 650:671 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 1 2 theme C. 231:232 arg1 difficile 234:242 C. difficile 231:242 C. difficile 231:242 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 3 3 from end 740:742 arg1 group 699:703 a 3-aminopropyl group 683:703 a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications 683:872 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 2 4 from α-linkage 407:415 arg1 unit 471:474 the repeating unit 457:474 the repeating unit 457:474 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 3 5 theme biological 838:847 arg1 studies 849:855 various biological studies 830:855 various biological studies 830:855 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 1 6 theme core 304:307 arg1 trisaccharide 309:321 the core trisaccharide 300:321 the core trisaccharide 300:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 2 7 theme azidosugars 519:529 arg1 imidates 507:514 glycosyl imidates 498:514 glycosyl imidates of azidosugars as donors 498:539 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 3 8 theme other 779:783 arg1 biomolecules 785:796 other biomolecules 779:796 other biomolecules 779:796 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 0 9 theme Surface 77:83 arg1 Polysaccharide 85:98 A Cell Surface Polysaccharide 70:98 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 0:126 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 0 10 theme Clostridium 105:115 arg1 difficile 117:125 Clostridium difficile 105:125 Clostridium difficile 105:125 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 3 11 theme 3-aminopropyl 685:697 arg1 group 699:703 a 3-aminopropyl group 683:703 a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications 683:872 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 2 12 theme phosphodiester 552:565 arg1 bridges 567:573 the phosphodiester bridges 548:573 the phosphodiester bridges between the oligosaccharides 548:602 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 2 13 theme H-phosphonate 625:637 arg1 chemistry 639:647 H-phosphonate chemistry 625:647 H-phosphonate chemistry 625:647 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 1 14 contain have 251:254 arg2 units 277:281 one or two repeating units 256:281 one or two repeating units of LTA linked to the core trisaccharide 256:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 14 contain have 251:254 arg1 glycans 169:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 15 theme lipoteichoic 180:191 arg1 acid 193:196 lipoteichoic acid 180:196 lipoteichoic acid (LTA, also known as PS-III) 180:224 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 16 theme acid 193:196 arg1 glycans 169:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 3 17 theme core 712:715 arg1 end 740:742 the core trisaccharide reducing end 708:742 the core trisaccharide reducing end 708:742 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 1 18 dep defined 145:151 arg1 functionalized 154:167 functionalized 154:167 functionalized 154:167 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 0 19 theme Defined 13:19 arg1 Glycans 40:46 Defined and Functionalized Glycans 13:46 Defined and Functionalized Glycans of Lipoteichoic Acid 13:67 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 0 20 theme Glycans 40:46 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 0:126 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 2 21 theme repeating 461:469 arg1 unit 471:474 the repeating unit 457:474 the repeating unit 457:474 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 1 22 dep acid 193:196 arg1 known 209:213 known 209:213 known as PS-III 209:223 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 0 23 theme Functionalized 25:38 arg1 Glycans 40:46 Defined and Functionalized Glycans 13:46 Defined and Functionalized Glycans of Lipoteichoic Acid 13:67 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 1 24 theme repeating 267:275 arg1 units 277:281 one or two repeating units 256:281 one or two repeating units of LTA linked to the core trisaccharide 256:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 25 from difficile 234:242 arg1 glycans 169:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 3 26 theme reducing 731:738 arg1 end 740:742 the core trisaccharide reducing end 708:742 the core trisaccharide reducing end 708:742 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 0 27 theme Lipoteichoic 51:62 arg1 Acid 64:67 Lipoteichoic Acid 51:67 Lipoteichoic Acid 51:67 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 1 28 theme convergent 359:368 arg1 [2 370:371 a convergent [2 357:371 a convergent [2 357:371 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 2 29 theme residues 445:452 arg1 α-linkage 407:415 The α-linkage 403:415 The α-linkage of both N-acetylglucosamine residues in the repeating unit 403:474 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 3 30 contain contained 673:681 arg2 group 699:703 a 3-aminopropyl group 683:703 a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications 683:872 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 3 30 contain contained 673:681 arg1 targets 665:671 Both synthetic targets 650:671 Both synthetic targets 650:671 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 3 31 theme trisaccharide 717:729 arg1 end 740:742 the core trisaccharide reducing end 708:742 the core trisaccharide reducing end 708:742 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 2 32 theme glycosyl 498:505 arg1 imidates 507:514 glycosyl imidates 498:514 glycosyl imidates of azidosugars as donors 498:539 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 1 33 attach linked 290:295 arg2 LTA 286:288 LTA 286:288 LTA linked to the core trisaccharide 286:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 33 attach linked 290:295 arg1 trisaccharide 309:321 the core trisaccharide 300:321 the core trisaccharide 300:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 1 34 theme LTA 286:288 arg1 units 277:281 one or two repeating units 256:281 one or two repeating units of LTA linked to the core trisaccharide 256:321 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 3 35 theme various 830:836 arg1 studies 849:855 various biological studies 830:855 various biological studies 830:855 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 1 36 dep known 209:213 arg1 LTA 199:201 LTA 199:201 LTA 199:201 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 2 37 theme N-acetylglucosamine 425:443 arg1 residues 445:452 both N-acetylglucosamine residues 420:452 both N-acetylglucosamine residues 420:452 The α-linkage of both N-acetylglucosamine residues in the repeating unit were constructed with glycosyl imidates of azidosugars as donors, while the phosphodiester bridges between the oligosaccharides were fashioned using H-phosphonate chemistry. 28548838 1 38 theme defined 145:151 arg1 glycans 169:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans 128:175 Two structurally defined, functionalized glycans of lipoteichoic acid (LTA, also known as PS-III) from C. difficile, which have one or two repeating units of LTA linked to the core trisaccharide, were efficiently synthesized via a convergent [2 + 3] or [2 + 2 + 3] strategy. 28548838 3 39 theme useful 819:824 arg1 conjugates 808:817 conjugates 808:817 conjugates useful for various biological studies and applications 808:872 Both synthetic targets contained a 3-aminopropyl group at the core trisaccharide reducing end, facilitating their conjugation to other biomolecules to afford conjugates useful for various biological studies and applications. 28548838 0 40 theme Acid 64:67 arg1 Glycans 40:46 Defined and Functionalized Glycans 13:46 Defined and Functionalized Glycans of Lipoteichoic Acid 13:67 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 0 41 dep Synthesis 0:8 arg1 Polysaccharide 85:98 A Cell Surface Polysaccharide 70:98 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 0:126 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 28548838 0 42 from difficile 117:125 arg1 Polysaccharide 85:98 A Cell Surface Polysaccharide 70:98 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 0:126 Synthesis of Defined and Functionalized Glycans of Lipoteichoic Acid: A Cell Surface Polysaccharide from Clostridium difficile. 29146600 3 0 theme Bioresource 639:649 arg1 Study 651:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 1 1 gly glycoproteins 159:171 arg1 glycoproteins 159:171 circulating glycoproteins 147:171 circulating glycoproteins 147:171 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 2 2 theme type 336:339 arg1 diabetes 343:350 type 1 diabetes 336:350 type 1 diabetes 336:350 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 9 3 theme growth 1659:1664 arg1 pathways 1675:1682 the epidermal growth factor receptor and transforming growth factor-β pathways 1605:1682 the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD 1605:1709 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 8 4 theme simpler 1427:1433 arg1 N-glycans 1435:1443 the simpler N-glycans 1423:1443 the simpler N-glycans (all P < 3.79 × 10-4) 1423:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 8 4 theme simpler 1427:1433 arg1 × 1459:1459 all P < 3.79 × 10-4 1446:1464 all P < 3.79 × 10-4 1446:1464 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 7 5 theme < 1293:1293 arg1 × 1300:1300 all P < 3.79 × 10-4 1287:1305 all P < 3.79 × 10-4 1287:1305 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 2 6 from function 324:331 arg1 diabetes 343:350 type 1 diabetes 336:350 type 1 diabetes 336:350 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 4 7 dep total 752:756 arg1 N-glycans 780:788 N-glycans 780:788 N-glycans 780:788 This yielded a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans. 29146600 7 8 with structures 1177:1186 arg1 branching 1198:1206 more branching 1193:1206 more branching 1193:1206 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 8 with structures 1177:1186 arg1 GP12 1243:1246 GP12, 26, 31, 32, and 34, and IGP19 and 23 1243:1284 GP12, 26, 31, 32, and 34, and IGP19 and 23 1243:1284 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 8 with structures 1177:1186 arg1 galactosylation 1209:1223 galactosylation 1209:1223 galactosylation 1209:1223 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 8 with structures 1177:1186 arg1 sialylation 1230:1240 sialylation 1230:1240 sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1230:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 2 9 theme glycemic 297:304 arg1 control 306:312 glycemic control 297:312 glycemic control 297:312 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 5 10 theme regression 798:807 arg1 models 809:814 Linear regression models 791:814 Linear regression models 791:814 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 11 11 theme altered 1811:1817 arg1 N-glycans 1819:1827 altered N-glycans 1811:1827 altered N-glycans 1811:1827 These data suggest that the role of altered N-glycans in DKD warrants further investigation. 29146600 3 12 theme N-glycan 693:700 arg1 profiles 702:709 total and IgG-specific N-glycan profiles 670:709 total and IgG-specific N-glycan profiles 670:709 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 6 13 theme type 991:994 arg1 diabetes 998:1005 type 1 diabetes 991:1005 type 1 diabetes 991:1005 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 7 14 dep GP12 1243:1246 arg1 × 1300:1300 all P < 3.79 × 10-4 1287:1305 all P < 3.79 × 10-4 1287:1305 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 9 15 theme CONCLUSIONS 1468:1478 arg1 HbA1c 1487:1491 CONCLUSIONS Higher HbA1c 1468:1491 CONCLUSIONS Higher HbA1c in type 1 diabetes 1468:1510 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 5 16 used used 821:824 arg2 models 809:814 Linear regression models 791:814 Linear regression models 791:814 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 3 17 contain have 440:443 arg1 adults 410:415 818 adults 406:415 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records 406:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 17 contain have 440:443 arg2 loss 460:463 extreme annual loss 445:463 extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) 445:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 18 theme clinical 548:555 arg1 records 557:563 retrospective clinical records 534:563 retrospective clinical records 534:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 9 19 theme type 1496:1499 arg1 diabetes 1503:1510 type 1 diabetes 1496:1510 type 1 diabetes 1496:1510 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 5 20 theme N-glycan 862:869 arg1 structures 871:880 N-glycan structures 862:880 N-glycan structures 862:880 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 11 21 from role 1803:1806 arg1 DKD 1832:1834 DKD 1832:1834 DKD 1832:1834 These data suggest that the role of altered N-glycans in DKD warrants further investigation. 29146600 7 22 theme biantennary 1107:1117 arg1 N-glycans 1119:1127 simple biantennary N-glycans 1100:1127 simple biantennary N-glycans 1100:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 1 23 theme circulating 147:157 arg1 glycoproteins 159:171 circulating glycoproteins 147:171 circulating glycoproteins 147:171 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 2 24 from control 306:312 arg1 diabetes 343:350 type 1 diabetes 336:350 type 1 diabetes 336:350 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 10 25 dep ACR 1755:1757 arg1 slope 1768:1772 slope 1768:1772 slope 1768:1772 Furthermore, N-glycans are associated with ACR and eGFR slope. 29146600 3 26 dep Using 381:385 arg1 from 566:569 from 566:569 from 566:569 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 7 27 theme relative 1078:1085 arg1 abundance 1087:1095 a lower relative abundance 1070:1095 a lower relative abundance of simple biantennary N-glycans 1070:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 1 28 theme Poorer 66:71 arg1 control 82:88 OBJECTIVE Poorer glycemic control 56:88 OBJECTIVE Poorer glycemic control in type 1 diabetes 56:107 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 0 29 from Disease 28:34 arg1 Diabetes 46:53 Type 1 Diabetes 39:53 Type 1 Diabetes 39:53 N-Glycan Profile and Kidney Disease in Type 1 Diabetes. 29146600 3 30 from adults 583:588 arg1 Study 651:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 31 theme 818 406:408 arg1 adults 410:415 818 adults 406:415 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records 406:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 0 32 theme N-Glycan 0:7 arg1 Profile 9:15 N-Glycan Profile 0:15 N-Glycan Profile 0:15 N-Glycan Profile and Kidney Disease in Type 1 Diabetes. 29146600 3 33 theme RESEARCH 353:360 arg1 DESIGN 362:367 RESEARCH DESIGN 353:367 RESEARCH DESIGN 353:367 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 8 34 theme greater 1348:1354 arg1 loss 1368:1371 greater mean annual loss 1348:1371 greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4) 1348:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 1 35 theme type 93:96 arg1 diabetes 100:107 type 1 diabetes 93:107 type 1 diabetes 93:107 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 8 36 theme annual 1361:1366 arg1 loss 1368:1371 greater mean annual loss 1348:1371 greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4) 1348:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 3 37 theme Diabetes 606:613 arg1 Type 632:635 the Scottish Diabetes Research Network Type 1 593:637 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 9 38 from changes 1531:1537 arg1 N-glycome 1552:1560 the serum N-glycome 1542:1560 the serum N-glycome 1542:1560 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 7 39 theme more 1193:1196 arg1 branching 1198:1206 more branching 1193:1206 more branching 1193:1206 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 0 40 theme Type 39:42 arg1 Diabetes 46:53 Type 1 Diabetes 39:53 Type 1 Diabetes 39:53 N-Glycan Profile and Kidney Disease in Type 1 Diabetes. 29146600 6 41 theme serum 1018:1022 arg1 IgG 1024:1026 total serum IgG 1012:1026 total serum IgG 1012:1026 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 3 42 theme annual 453:458 arg1 loss 460:463 extreme annual loss 445:463 extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) 445:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 9 43 theme growth 1619:1624 arg1 receptor 1633:1640 the epidermal growth factor receptor 1605:1640 the epidermal growth factor receptor 1605:1640 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 3 44 theme Network 624:630 arg1 Type 632:635 the Scottish Diabetes Research Network Type 1 593:637 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 45 from adults 410:415 arg1 samples 393:399 serum samples 387:399 serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records 387:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 1 46 from control 82:88 arg1 diabetes 100:107 type 1 diabetes 93:107 type 1 diabetes 93:107 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 1 47 theme kidney 224:229 arg1 DKD 240:242 DKD 240:242 DKD 240:242 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 1 47 theme kidney 224:229 arg1 disease 231:237 diabetic kidney disease 215:237 diabetic kidney disease (DKD) 215:243 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 5 48 theme eGFR 932:935 arg1 slope 937:941 eGFR slope 932:941 eGFR slope 932:941 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 6 49 theme diabetes 998:1005 arg1 age 969:971 age 969:971 age 969:971 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 6 49 theme diabetes 998:1005 arg1 duration 979:986 duration 979:986 duration of type 1 diabetes 979:1005 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 6 49 theme diabetes 998:1005 arg1 sex 974:976 sex 974:976 sex 974:976 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 6 49 theme diabetes 998:1005 arg1 IgG 1024:1026 total serum IgG 1012:1026 total serum IgG 1012:1026 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 9 50 theme receptor 1633:1640 arg1 pathways 1675:1682 the epidermal growth factor receptor and transforming growth factor-β pathways 1605:1682 the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD 1605:1709 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 3 51 theme glomerular 478:487 arg1 eGFR 506:509 eGFR 506:509 eGFR 506:509 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 51 theme glomerular 478:487 arg1 rate 500:503 estimated glomerular filtration rate 468:503 estimated glomerular filtration rate (eGFR; i.e., slope) 468:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 8 52 theme < 1452:1452 arg1 N-glycans 1435:1443 the simpler N-glycans 1423:1443 the simpler N-glycans (all P < 3.79 × 10-4) 1423:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 8 52 theme < 1452:1452 arg1 × 1459:1459 all P < 3.79 × 10-4 1446:1464 all P < 3.79 × 10-4 1446:1464 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 7 53 theme higher 1135:1140 arg1 abundance 1151:1159 a higher relative abundance 1133:1159 a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1133:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 9 54 theme transforming 1646:1657 arg1 growth 1659:1664 transforming growth 1646:1664 transforming growth 1646:1664 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 7 55 theme P 1291:1291 arg1 × 1300:1300 all P < 3.79 × 10-4 1287:1305 all P < 3.79 × 10-4 1287:1305 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 9 56 theme factor-β 1666:1673 arg1 pathways 1675:1682 the epidermal growth factor receptor and transforming growth factor-β pathways 1605:1682 the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD 1605:1709 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 3 57 theme IgG-specific 680:691 arg1 profiles 702:709 total and IgG-specific N-glycan profiles 670:709 total and IgG-specific N-glycan profiles 670:709 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 4 58 theme IgG 770:772 arg1 abundance 736:744 a relative abundance 725:744 a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans 725:788 This yielded a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans. 29146600 3 59 theme total 670:674 arg1 profiles 702:709 total and IgG-specific N-glycan profiles 670:709 total and IgG-specific N-glycan profiles 670:709 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 60 from loss 460:463 arg1 eGFR 506:509 eGFR 506:509 eGFR 506:509 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 60 from loss 460:463 arg1 rate 500:503 estimated glomerular filtration rate 468:503 estimated glomerular filtration rate (eGFR; i.e., slope) 468:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 5 61 theme Linear 791:796 arg1 models 809:814 Linear regression models 791:814 Linear regression models 791:814 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 11 62 theme N-glycans 1819:1827 arg1 role 1803:1806 the role 1799:1806 the role of altered N-glycans in DKD 1799:1834 These data suggest that the role of altered N-glycans in DKD warrants further investigation. 29146600 7 63 theme structures 1177:1186 arg1 abundance 1087:1095 a lower relative abundance 1070:1095 a lower relative abundance of simple biantennary N-glycans 1070:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 63 theme structures 1177:1186 arg1 abundance 1151:1159 a higher relative abundance 1133:1159 a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1133:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 9 64 theme Higher 1480:1485 arg1 HbA1c 1487:1491 CONCLUSIONS Higher HbA1c 1468:1491 CONCLUSIONS Higher HbA1c in type 1 diabetes 1468:1510 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 3 65 theme retrospective 534:546 arg1 records 557:563 retrospective clinical records 534:563 retrospective clinical records 534:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 66 dep eGFR 506:509 arg1 i.e. 512:515 i.e. 512:515 i.e. 512:515 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 66 dep eGFR 506:509 arg1 slope 518:522 slope 518:522 slope 518:522 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 11 67 theme further 1845:1851 arg1 investigation 1853:1865 further investigation 1845:1865 further investigation 1845:1865 These data suggest that the role of altered N-glycans in DKD warrants further investigation. 29146600 2 68 theme renal 318:322 arg1 function 324:331 renal function 318:331 renal function 318:331 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 7 69 theme N-glycans 1119:1127 arg1 abundance 1087:1095 a lower relative abundance 1070:1095 a lower relative abundance of simple biantennary N-glycans 1070:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 69 theme N-glycans 1119:1127 arg1 abundance 1151:1159 a higher relative abundance 1133:1159 a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1133:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 1 70 theme N-glycosylation 119:133 arg1 patterns 135:142 N-glycosylation patterns 119:142 N-glycosylation patterns on circulating glycoproteins 119:171 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 9 71 from HbA1c 1487:1491 arg1 diabetes 1503:1510 type 1 diabetes 1496:1510 type 1 diabetes 1496:1510 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 8 72 theme Similar 1309:1315 arg1 patterns 1317:1324 Similar patterns 1309:1324 Similar patterns 1309:1324 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 7 73 theme simple 1100:1105 arg1 N-glycans 1119:1127 simple biantennary N-glycans 1100:1127 simple biantennary N-glycans 1100:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 2 74 from N-glycans 283:291 arg1 diabetes 343:350 type 1 diabetes 336:350 type 1 diabetes 336:350 We investigated associations between N-glycans and glycemic control and renal function in type 1 diabetes. 29146600 0 75 from Profile 9:15 arg1 Diabetes 46:53 Type 1 Diabetes 39:53 Type 1 Diabetes 39:53 N-Glycan Profile and Kidney Disease in Type 1 Diabetes. 29146600 5 76 theme albumin-to-creatinine 893:913 arg1 ratio 915:919 albumin-to-creatinine ratio 893:919 albumin-to-creatinine ratio (ACR) 893:925 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 5 76 theme albumin-to-creatinine 893:913 arg1 ACR 922:924 ACR 922:924 ACR 922:924 Linear regression models were used to investigate associations between N-glycan structures and HbA1c, albumin-to-creatinine ratio (ACR), and eGFR slope. 29146600 7 77 theme lower 1072:1076 arg1 abundance 1087:1095 a lower relative abundance 1070:1095 a lower relative abundance of simple biantennary N-glycans 1070:1127 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 7 78 gly sialylation 1230:1240 arg1 structures 1177:1186 more complex structures 1164:1186 more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1164:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 1 79 from patterns 135:142 arg1 glycoproteins 159:171 circulating glycoproteins 147:171 circulating glycoproteins 147:171 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 1 80 theme OBJECTIVE 56:64 arg1 control 82:88 OBJECTIVE Poorer glycemic control 56:88 OBJECTIVE Poorer glycemic control in type 1 diabetes 56:107 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 9 81 theme serum 1546:1550 arg1 N-glycome 1552:1560 the serum N-glycome 1542:1560 the serum N-glycome 1542:1560 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 7 82 dep RESULTS 1029:1035 arg1 associated 1054:1063 associated 1054:1063 was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1050:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 4 83 theme relative 727:734 arg1 abundance 736:744 a relative abundance 725:744 a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans 725:788 This yielded a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans. 29146600 1 84 theme glycemic 73:80 arg1 control 82:88 OBJECTIVE Poorer glycemic control 56:88 OBJECTIVE Poorer glycemic control in type 1 diabetes 56:107 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 8 85 theme mean 1356:1359 arg1 loss 1368:1371 greater mean annual loss 1348:1371 greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4) 1348:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 0 86 theme Kidney 21:26 arg1 Disease 28:34 Kidney Disease 21:34 Kidney Disease 21:34 N-Glycan Profile and Kidney Disease in Type 1 Diabetes. 29146600 3 87 theme Scottish 597:604 arg1 Type 632:635 the Scottish Diabetes Research Network Type 1 593:637 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 88 theme extreme 445:451 arg1 loss 460:463 extreme annual loss 445:463 extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) 445:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 7 89 theme Higher 1037:1042 arg1 HbA1c 1044:1048 Higher HbA1c 1037:1048 Higher HbA1c 1037:1048 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 8 90 theme eGFR 1376:1379 arg1 loss 1368:1371 greater mean annual loss 1348:1371 greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4) 1348:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 8 90 theme eGFR 1376:1379 arg1 ACR 1340:1342 ACR 1340:1342 ACR 1340:1342 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 7 91 theme complex 1169:1175 arg1 structures 1177:1186 more complex structures 1164:1186 more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1164:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 3 92 theme serum 387:391 arg1 samples 393:399 serum samples 387:399 serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records 387:563 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 93 theme Type 632:635 arg1 Study 651:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 94 theme filtration 489:498 arg1 eGFR 506:509 eGFR 506:509 eGFR 506:509 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 94 theme filtration 489:498 arg1 rate 500:503 estimated glomerular filtration rate 468:503 estimated glomerular filtration rate (eGFR; i.e., slope) 468:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 95 theme estimated 468:476 arg1 eGFR 506:509 eGFR 506:509 eGFR 506:509 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 3 95 theme estimated 468:476 arg1 rate 500:503 estimated glomerular filtration rate 468:503 estimated glomerular filtration rate (eGFR; i.e., slope) 468:523 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 6 96 theme total 1012:1016 arg1 IgG 1024:1026 total serum IgG 1012:1026 total serum IgG 1012:1026 Models were adjusted for age, sex, duration of type 1 diabetes, and total serum IgG. 29146600 9 97 theme epidermal 1609:1617 arg1 receptor 1633:1640 the epidermal growth factor receptor 1605:1640 the epidermal growth factor receptor 1605:1640 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 3 98 theme Research 615:622 arg1 Type 632:635 the Scottish Diabetes Research Network Type 1 593:637 the Scottish Diabetes Research Network Type 1 Bioresource Study 593:655 RESEARCH DESIGN AND METHODS Using serum samples from 818 adults who were considered to have extreme annual loss in estimated glomerular filtration rate (eGFR; i.e., slope) based on retrospective clinical records, from among 6,127 adults in the Scottish Diabetes Research Network Type 1 Bioresource Study, we measured total and IgG-specific N-glycan profiles. 29146600 8 99 theme P 1450:1450 arg1 N-glycans 1435:1443 the simpler N-glycans 1423:1443 the simpler N-glycans (all P < 3.79 × 10-4) 1423:1465 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 8 99 theme P 1450:1450 arg1 × 1459:1459 all P < 3.79 × 10-4 1446:1464 all P < 3.79 × 10-4 1446:1464 Similar patterns were seen for ACR and greater mean annual loss of eGFR, which were also associated with fewer of the simpler N-glycans (all P < 3.79 × 10-4). 29146600 1 100 theme diabetic 215:222 arg1 DKD 240:242 DKD 240:242 DKD 240:242 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 1 100 theme diabetic 215:222 arg1 disease 231:237 diabetic kidney disease 215:237 diabetic kidney disease (DKD) 215:243 OBJECTIVE Poorer glycemic control in type 1 diabetes may alter N-glycosylation patterns on circulating glycoproteins, and these alterations may be linked with diabetic kidney disease (DKD). 29146600 7 101 theme relative 1142:1149 arg1 abundance 1151:1159 a higher relative abundance 1133:1159 a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4) 1133:1306 RESULTS Higher HbA1c was associated with a lower relative abundance of simple biantennary N-glycans and a higher relative abundance of more complex structures with more branching, galactosylation, and sialylation (GP12, 26, 31, 32, and 34, and IGP19 and 23; all P < 3.79 × 10-4). 29146600 9 102 theme factor 1626:1631 arg1 receptor 1633:1640 the epidermal growth factor receptor 1605:1640 the epidermal growth factor receptor 1605:1640 CONCLUSIONS Higher HbA1c in type 1 diabetes is associated with changes in the serum N-glycome that have elsewhere been shown to regulate the epidermal growth factor receptor and transforming growth factor-β pathways that are implicated in DKD. 29146600 4 103 theme total 752:756 arg1 abundance 736:744 a relative abundance 725:744 a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans 725:788 This yielded a relative abundance of 39 total (GP) and 24 IgG (IGP) N-glycans. 26512077 11 0 theme virus 2815:2819 arg1 infection 2821:2829 virus infection 2815:2829 virus infection 2815:2829 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 6 1 theme TfRs 1314:1317 arg1 Analysis 1298:1305 Analysis 1298:1305 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies 1298:1379 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 8 2 dep emergence 1855:1863 arg1 the 1851:1853 the 1851:1853 the 1851:1853 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 7 3 theme different 1673:1681 arg1 hosts 1693:1697 different carnivore hosts 1673:1697 different carnivore hosts 1673:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 9 4 from region 2055:2060 arg1 structure 2076:2084 the capsid structure 2065:2084 the capsid structure of CPV 2065:2091 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 9 5 theme hosts 2234:2238 arg1 infection 2195:2203 infection 2195:2203 infection of previously nonsusceptible hosts 2195:2238 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 10 6 theme parvovirus 2321:2330 arg1 capsid 2332:2337 the parvovirus capsid 2317:2337 the parvovirus capsid 2317:2337 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 3 7 theme acid 614:617 arg1 variation 619:627 extensive amino acid variation 598:627 extensive amino acid variation among the carnivore parvoviruses 598:660 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 5 8 theme new 1269:1271 arg1 residue 1273:1279 the new residue 1265:1279 the new residue at position 300 1265:1295 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 2 9 theme prototype 515:523 arg1 CPV 533:535 the prototype raccoon CPV 511:535 the prototype raccoon CPV 511:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 3 10 theme carnivore 639:647 arg1 parvoviruses 649:660 the carnivore parvoviruses 635:660 the carnivore parvoviruses 635:660 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 0 11 theme Capsid 96:101 arg1 Spike 62:66 the Three-Fold Spike 47:66 the Three-Fold Spike of the Carnivore Parvovirus Capsid 47:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 6 12 theme evolution 1363:1371 arg1 studies 1373:1379 the experimental evolution studies 1346:1379 the experimental evolution studies 1346:1379 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 5 13 from passage 1054:1060 arg1 hosts 1075:1079 different hosts 1065:1079 different hosts 1065:1079 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 7 14 theme range 1824:1828 arg1 determinant 1804:1814 a key determinant 1798:1814 a key determinant of host range 1798:1828 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 14 theme range 1824:1828 arg1 position 1782:1789 VP2 position 300 1778:1793 VP2 position 300 1778:1793 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 2 15 theme VP2 490:492 arg1 position 495:502 capsid (VP2) position 300 482:506 capsid (VP2) position 300 in the prototype raccoon CPV 482:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 3 16 from mutants 829:835 arg1 clones 851:856 infectious clones 840:856 infectious clones 840:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 10 17 from residue 2306:2312 arg1 nature 2342:2347 nature 2342:2347 nature 2342:2347 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 10 17 from residue 2306:2312 arg1 capsid 2332:2337 the parvovirus capsid 2317:2337 the parvovirus capsid 2317:2337 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 7 18 theme alternative 1615:1625 arg1 position 1627:1634 alternative position 1615:1634 alternative position 300 residues to bind TfRs and infect different carnivore hosts 1615:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 2 19 theme capsid 482:487 arg1 position 495:502 capsid (VP2) position 300 482:506 capsid (VP2) position 300 in the prototype raccoon CPV 482:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 3 20 theme host 711:714 arg1 range 716:720 host range 711:720 host range 711:720 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 7 21 theme viruses 1602:1608 arg1 abilities 1589:1597 the abilities 1585:1597 the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts 1585:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 2 22 theme single 424:429 arg1 mutation 431:438 a single mutation 422:438 a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV 422:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 1 23 theme pandemic 275:282 arg1 parvovirus 255:264 canine parvovirus 248:264 canine parvovirus (CPV) 248:270 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 23 theme pandemic 275:282 arg1 pathogen 284:291 a pandemic pathogen 273:291 a pandemic pathogen of domestic dogs 273:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 9 24 theme capsid 2069:2074 arg1 structure 2076:2084 the capsid structure 2065:2084 the capsid structure of CPV 2065:2091 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 5 25 theme cell 1041:1044 arg1 passage 1054:1060 cell culture passage 1041:1060 cell culture passage in different hosts 1041:1079 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 10 26 theme critical 2374:2381 arg1 it 2366:2367 it 2366:2367 it 2366:2367 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 10 26 theme critical 2374:2381 arg1 determinant 2383:2393 a critical determinant 2372:2393 a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection 2372:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 0 27 theme Three-Fold 51:60 arg1 Spike 62:66 the Three-Fold Spike 47:66 the Three-Fold Spike of the Carnivore Parvovirus Capsid 47:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 7 28 theme host 1819:1822 arg1 range 1824:1828 host range 1819:1828 host range 1819:1828 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 1 29 theme Sylvatic 140:147 arg1 carnivores 149:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 29 theme Sylvatic 140:147 arg1 raccoons 169:176 raccoons 169:176 raccoons 169:176 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 29 theme Sylvatic 140:147 arg1 hosts 222:226 important hosts 212:226 important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs 212:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 2 30 theme aspartic 452:459 arg1 acid 461:464 an aspartic acid 449:464 an aspartic acid 449:464 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 2 31 theme cell 548:551 arg1 infection 553:561 dog cell infection 544:561 dog cell infection 544:561 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 1 32 theme dogs 305:308 arg1 parvovirus 255:264 canine parvovirus 248:264 canine parvovirus (CPV) 248:270 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 32 theme dogs 305:308 arg1 pathogen 284:291 a pandemic pathogen 273:291 a pandemic pathogen of domestic dogs 273:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 7 33 theme significant 1558:1568 arg1 differences 1570:1580 significant differences 1558:1580 significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts 1558:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 34 dep position 1627:1634 arg1 residues 1640:1647 300 residues 1636:1647 alternative position 300 residues to bind TfRs and infect different carnivore hosts 1615:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 10 35 theme cross-species 2402:2414 arg1 transfer 2416:2423 the cross-species transfer 2398:2423 the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection 2398:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 5 36 theme hosts 1190:1194 arg1 infections 1164:1173 natural infections 1156:1173 natural infections of alternative hosts 1156:1194 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 0 37 theme Carnivore 75:83 arg1 Capsid 96:101 the Carnivore Parvovirus Capsid 71:101 the Carnivore Parvovirus Capsid 71:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 5 38 theme residues 978:985 arg1 Changes 958:964 Changes 958:964 Changes of adjacent residues (residues 299 and 301) 958:1008 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 10 39 theme variable 2297:2304 arg1 position 2272:2279 VP2 position 300 2268:2283 VP2 position 300 2268:2283 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 10 39 theme variable 2297:2304 arg1 residue 2306:2312 the most variable residue 2288:2312 the most variable residue in the parvovirus capsid in nature 2288:2347 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 3 40 theme comprehensive 791:803 arg1 mutants 829:835 VP2 position 300 mutants 812:835 VP2 position 300 mutants in infectious clones 812:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 40 theme comprehensive 791:803 arg1 set 805:807 a comprehensive set 789:807 a comprehensive set of VP2 position 300 mutants in infectious clones 789:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 4 41 theme dog 926:928 arg1 cells 951:955 dog, but not cat or fox, cells 926:955 dog, but not cat or fox, cells 926:955 Notably, some position 300 residues rendered CPV noninfectious for dog, but not cat or fox, cells. 26512077 0 42 theme Single 0:5 arg1 Mutations 7:15 Single Mutations 0:15 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid 0:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 6 43 theme domestic 1475:1482 arg1 TfR 1488:1490 the domestic dog TfR 1471:1490 the domestic dog TfR 1471:1490 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 3 44 theme mutants 829:835 arg1 mutants 829:835 VP2 position 300 mutants 812:835 VP2 position 300 mutants in infectious clones 812:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 44 theme mutants 829:835 arg1 set 805:807 a comprehensive set 789:807 a comprehensive set of VP2 position 300 mutants in infectious clones 789:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 10 45 theme VP2 2268:2270 arg1 position 2272:2279 VP2 position 300 2268:2283 VP2 position 300 2268:2283 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 10 45 theme VP2 2268:2270 arg1 residue 2306:2312 the most variable residue 2288:2312 the most variable residue in the parvovirus capsid in nature 2288:2347 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 10 46 from determinant 2383:2393 arg1 transfer 2416:2423 the cross-species transfer 2398:2423 the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection 2398:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 11 47 theme carnivore 2695:2703 arg1 species 2705:2711 different carnivore species 2685:2711 different carnivore species 2685:2711 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 5 48 theme natural 1156:1162 arg1 infections 1164:1173 natural infections 1156:1173 natural infections of alternative hosts 1156:1194 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 2 49 theme dog 370:372 arg1 TfR 396:398 TfR 396:398 TfR 396:398 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 2 49 theme dog 370:372 arg1 receptor 386:393 the dog transferrin receptor 366:393 the dog transferrin receptor (TfR) 366:399 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 8 50 theme canine 1888:1893 arg1 CPV 1907:1909 CPV 1907:1909 CPV 1907:1909 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 8 50 theme canine 1888:1893 arg1 parvovirus 1895:1904 canine parvovirus 1888:1904 canine parvovirus (CPV) 1888:1910 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 1 51 theme parvovirus 255:264 arg1 evolution 235:243 the evolution 231:243 the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs 231:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 7 52 theme VP2 1778:1780 arg1 determinant 1804:1814 a key determinant 1798:1814 a key determinant of host range 1798:1828 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 52 theme VP2 1778:1780 arg1 position 1782:1789 VP2 position 300 1778:1793 VP2 position 300 1778:1793 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 3 53 theme position 816:823 arg1 mutants 829:835 VP2 position 300 mutants 812:835 VP2 position 300 mutants in infectious clones 812:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 54 from clones 851:856 arg1 mutants 829:835 VP2 position 300 mutants 812:835 VP2 position 300 mutants in infectious clones 812:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 54 from clones 851:856 arg1 set 805:807 a comprehensive set 789:807 a comprehensive set of VP2 position 300 mutants in infectious clones 789:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 2 55 theme dog 411:413 arg1 cells 415:419 dog cells 411:419 dog cells 411:419 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 0 56 theme VP2 24:26 arg1 Region 37:42 the VP2 300 Loop Region 20:42 the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid 20:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 2 57 from raccoons 333:340 arg1 viruses 320:326 viruses 320:326 viruses from raccoons 320:340 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 11 58 theme receptor 2612:2619 arg1 binding 2621:2627 receptor binding 2612:2627 receptor binding 2612:2627 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 9 59 theme carnivore 2165:2173 arg1 hosts 2175:2179 alternative carnivore hosts 2153:2179 alternative carnivore hosts 2153:2179 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 0 60 theme Loop 32:35 arg1 Region 37:42 the VP2 300 Loop Region 20:42 the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid 20:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 3 61 theme infectious 840:849 arg1 clones 851:856 infectious clones 840:856 infectious clones 840:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 5 62 from position 1284:1291 arg1 residue 1273:1279 the new residue 1265:1279 the new residue at position 300 1265:1295 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 11 63 theme VP2 2656:2658 arg1 mutants 2673:2679 various VP2 position 300 mutants 2648:2679 various VP2 position 300 mutants 2648:2679 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 5 64 theme compensatory 1213:1224 arg1 mutations 1226:1234 compensatory mutations 1213:1234 compensatory mutations 1213:1234 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 6 65 theme hosts 1332:1336 arg1 TfRs 1314:1317 the TfRs 1310:1317 the TfRs of carnivore hosts used in the experimental evolution studies 1310:1379 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 7 66 theme carnivore 1683:1691 arg1 hosts 1693:1697 different carnivore hosts 1673:1697 different carnivore hosts 1673:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 4 67 theme cat 939:941 arg1 cells 951:955 dog, but not cat or fox, cells 926:955 dog, but not cat or fox, cells 926:955 Notably, some position 300 residues rendered CPV noninfectious for dog, but not cat or fox, cells. 26512077 3 68 theme amino 608:612 arg1 variation 619:627 extensive amino acid variation 598:627 extensive amino acid variation among the carnivore parvoviruses 598:660 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 11 69 theme position 2660:2667 arg1 mutants 2673:2679 various VP2 position 300 mutants 2648:2679 various VP2 position 300 mutants 2648:2679 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 11 70 from mutations 2729:2737 arg1 region 2747:2752 this region 2742:2752 this region 2742:2752 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 9 71 theme nonsusceptible 2219:2232 arg1 hosts 2234:2238 previously nonsusceptible hosts 2208:2238 previously nonsusceptible hosts 2208:2238 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 5 72 theme different 1065:1073 arg1 hosts 1075:1079 different hosts 1065:1079 different hosts 1065:1079 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 8 73 dep hosts 1947:1951 arg1 the 1933:1935 the 1933:1935 the 1933:1935 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 6 74 theme glycosylation 1405:1417 arg1 patterns 1419:1426 their glycosylation patterns 1399:1426 their glycosylation patterns 1399:1426 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 2 75 theme raccoon 525:531 arg1 CPV 533:535 the prototype raccoon CPV 511:535 the prototype raccoon CPV 511:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 3 76 from evolution 753:761 arg1 nature 766:771 nature 766:771 nature 766:771 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 11 77 from differences 2597:2607 arg1 infectivity 2633:2643 infectivity 2633:2643 infectivity 2633:2643 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 11 77 from differences 2597:2607 arg1 binding 2621:2627 receptor binding 2612:2627 receptor binding 2612:2627 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 1 78 from hosts 222:226 arg1 evolution 235:243 the evolution 231:243 the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs 231:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 6 79 theme experimental 1350:1361 arg1 studies 1373:1379 the experimental evolution studies 1346:1379 the experimental evolution studies 1346:1379 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 11 80 theme substantial 2585:2595 arg1 differences 2597:2607 substantial differences 2585:2607 substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species 2585:2711 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 6 81 from present 1455:1461 arg1 TfR 1488:1490 the domestic dog TfR 1471:1490 the domestic dog TfR 1471:1490 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 6 82 theme glycan 1448:1453 arg1 present 1455:1461 a glycan present 1446:1461 a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses 1446:1535 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 6 83 dep varied 1428:1433 arg1 including 1436:1444 including 1436:1444 including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses 1436:1535 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 3 84 from set 805:807 arg1 clones 851:856 infectious clones 840:856 infectious clones 840:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 7 85 from differences 1570:1580 arg1 abilities 1589:1597 the abilities 1585:1597 the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts 1585:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 9 86 theme CPV 2089:2091 arg1 structure 2076:2084 the capsid structure 2065:2084 the capsid structure of CPV 2065:2091 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 0 87 theme Spike 62:66 arg1 Region 37:42 the VP2 300 Loop Region 20:42 the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid 20:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 1 88 theme UNLABELLED 129:138 arg1 carnivores 149:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 88 theme UNLABELLED 129:138 arg1 raccoons 169:176 raccoons 169:176 raccoons 169:176 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 88 theme UNLABELLED 129:138 arg1 hosts 222:226 important hosts 212:226 important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs 212:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 4 89 theme position 873:880 arg1 residues 886:893 some position 300 residues 868:893 some position 300 residues 868:893 Notably, some position 300 residues rendered CPV noninfectious for dog, but not cat or fox, cells. 26512077 0 90 from Mutations 7:15 arg1 Region 37:42 the VP2 300 Loop Region 20:42 the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid 20:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 1 91 theme domestic 296:303 arg1 dogs 305:308 domestic dogs 296:308 domestic dogs 296:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 7 92 theme key 1800:1802 arg1 determinant 1804:1814 a key determinant 1798:1814 a key determinant of host range 1798:1828 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 92 theme key 1800:1802 arg1 position 1782:1789 VP2 position 300 1778:1793 VP2 position 300 1778:1793 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 93 with viruses 1602:1608 arg1 position 1627:1634 alternative position 1615:1634 alternative position 300 residues to bind TfRs and infect different carnivore hosts 1615:1697 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 2 94 theme dog 544:546 arg1 infection 553:561 dog cell infection 544:561 dog cell infection 544:561 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 11 95 theme different 2685:2693 arg1 species 2705:2711 different carnivore species 2685:2711 different carnivore species 2685:2711 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 5 96 theme alternative 1178:1188 arg1 hosts 1190:1194 alternative hosts 1178:1194 alternative hosts 1178:1194 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 9 97 theme VP2 2110:2112 arg1 position 2114:2121 VP2 position 300 2110:2125 VP2 position 300 2110:2125 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 0 98 theme Parvovirus 85:94 arg1 Capsid 96:101 the Carnivore Parvovirus Capsid 71:101 the Carnivore Parvovirus Capsid 71:101 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 3 99 dep investigated 674:685 arg1 analyzing 725:733 analyzing 725:733 analyzing its diversity and evolution in nature 725:771 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 99 dep investigated 674:685 arg1 creating 780:787 creating 780:787 creating a comprehensive set of VP2 position 300 mutants in infectious clones 780:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 3 100 theme VP2 812:814 arg1 mutants 829:835 VP2 position 300 mutants 812:835 VP2 position 300 mutants in infectious clones 812:856 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 11 101 theme mutants 2673:2679 arg1 infectivity 2633:2643 infectivity 2633:2643 infectivity 2633:2643 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 11 101 theme mutants 2673:2679 arg1 binding 2621:2627 receptor binding 2612:2627 receptor binding 2612:2627 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 8 102 theme pandemic 1869:1876 arg1 spread 1878:1883 pandemic spread 1869:1883 pandemic spread 1869:1883 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 6 103 theme dog 1484:1486 arg1 TfR 1488:1490 the domestic dog TfR 1471:1490 the domestic dog TfR 1471:1490 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 5 104 theme adjacent 969:976 arg1 residues 978:985 adjacent residues 969:985 adjacent residues (residues 299 and 301) 969:1008 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 5 104 theme adjacent 969:976 arg1 residues 988:995 residues 299 and 301 988:1007 residues 299 and 301 988:1007 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 0 105 theme Host 117:120 arg1 Range 122:126 Host Range 117:126 Host Range 117:126 Single Mutations in the VP2 300 Loop Region of the Three-Fold Spike of the Carnivore Parvovirus Capsid Can Determine Host Range. 26512077 2 106 from position 495:502 arg1 glycine 471:477 a glycine 469:477 a glycine at capsid (VP2) position 300 in the prototype raccoon CPV 469:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 2 106 from position 495:502 arg1 CPV 533:535 the prototype raccoon CPV 511:535 the prototype raccoon CPV 511:535 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 10 107 theme different 2444:2452 arg1 carnivores 2454:2463 different carnivores 2444:2463 different carnivores due to its interactions with the transferrin receptor to mediate infection 2444:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 8 108 theme parvovirus 1895:1904 arg1 emergence 1855:1863 emergence 1855:1863 emergence 1855:1863 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 8 108 theme parvovirus 1895:1904 arg1 spread 1878:1883 pandemic spread 1869:1883 pandemic spread 1869:1883 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 1 109 theme canine 248:253 arg1 parvovirus 255:264 canine parvovirus 248:264 canine parvovirus (CPV) 248:270 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 109 theme canine 248:253 arg1 pathogen 284:291 a pandemic pathogen 273:291 a pandemic pathogen of domestic dogs 273:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 109 theme canine 248:253 arg1 CPV 267:269 CPV 267:269 CPV 267:269 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 10 110 with interactions 2476:2487 arg1 receptor 2510:2517 the transferrin receptor 2494:2517 the transferrin receptor 2494:2517 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 2 111 theme transferrin 374:384 arg1 TfR 396:398 TfR 396:398 TfR 396:398 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 2 111 theme transferrin 374:384 arg1 receptor 386:393 the dog transferrin receptor 366:393 the dog transferrin receptor (TfR) 366:399 Although viruses from raccoons do not efficiently bind the dog transferrin receptor (TfR) or infect dog cells, a single mutation changing an aspartic acid to a glycine at capsid (VP2) position 300 in the prototype raccoon CPV allows dog cell infection. 26512077 10 112 theme viruses 2428:2434 arg1 transfer 2416:2423 the cross-species transfer 2398:2423 the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection 2398:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 5 113 theme culture 1046:1052 arg1 passage 1054:1060 cell culture passage 1041:1060 cell culture passage in different hosts 1041:1079 Changes of adjacent residues (residues 299 and 301) were also observed often after cell culture passage in different hosts, and some of the mutations mimicked changes seen in viruses recovered from natural infections of alternative hosts, suggesting that compensatory mutations were selected to accommodate the new residue at position 300. 26512077 11 114 theme single 2722:2727 arg1 mutations 2729:2737 single mutations 2722:2737 single mutations in this region 2722:2752 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 8 115 theme evolutionary 1957:1968 arg1 pathways 1970:1977 evolutionary pathways 1957:1977 evolutionary pathways 1957:1977 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 3 116 from diversity 739:747 arg1 nature 766:771 nature 766:771 nature 766:771 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 8 117 theme carnivore 1937:1945 arg1 hosts 1947:1951 carnivore hosts 1937:1951 carnivore hosts 1937:1951 IMPORTANCE Although the emergence and pandemic spread of canine parvovirus (CPV) are well documented, the carnivore hosts and evolutionary pathways involved in its emergence remain enigmatic. 26512077 9 118 theme alternative 2153:2163 arg1 hosts 2175:2179 alternative carnivore hosts 2153:2179 alternative carnivore hosts 2153:2179 We recently demonstrated that a region in the capsid structure of CPV, centered around VP2 position 300, varies after transfer to alternative carnivore hosts and may allow infection of previously nonsusceptible hosts in vitro. 26512077 4 119 theme CPV 904:906 arg1 noninfectious 908:920 CPV noninfectious 904:920 CPV noninfectious for dog, but not cat or fox, cells 904:955 Notably, some position 300 residues rendered CPV noninfectious for dog, but not cat or fox, cells. 26512077 3 120 theme VP2 572:574 arg1 position 576:583 VP2 position 300 572:587 VP2 position 300 572:587 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 10 121 theme due 2465:2467 arg1 carnivores 2454:2463 different carnivores 2444:2463 different carnivores due to its interactions with the transferrin receptor to mediate infection 2444:2538 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 7 122 theme infection 1734:1742 arg1 process 1723:1729 the process 1719:1729 the process of infection 1719:1742 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 7 122 theme infection 1734:1742 arg1 dependent 1759:1767 dependent 1759:1767 dependent 1759:1767 Overall, there were significant differences in the abilities of viruses with alternative position 300 residues to bind TfRs and infect different carnivore hosts, demonstrating that the process of infection is highly host dependent and that VP2 position 300 is a key determinant of host range. 26512077 10 123 theme transferrin 2498:2508 arg1 receptor 2510:2517 the transferrin receptor 2494:2517 the transferrin receptor 2494:2517 Here we show that VP2 position 300 is the most variable residue in the parvovirus capsid in nature, suggesting that it is a critical determinant in the cross-species transfer of viruses between different carnivores due to its interactions with the transferrin receptor to mediate infection. 26512077 3 124 theme extensive 598:606 arg1 variation 619:627 extensive amino acid variation 598:627 extensive amino acid variation among the carnivore parvoviruses 598:660 Because VP2 position 300 exhibits extensive amino acid variation among the carnivore parvoviruses, we further investigated its role in determining host range by analyzing its diversity and evolution in nature and by creating a comprehensive set of VP2 position 300 mutants in infectious clones. 26512077 11 125 theme various 2648:2654 arg1 mutants 2673:2679 various VP2 position 300 mutants 2648:2679 various VP2 position 300 mutants 2648:2679 To this end, we demonstrated that there are substantial differences in receptor binding and infectivity of various VP2 position 300 mutants for different carnivore species and that single mutations in this region can influence whether a host is susceptible or refractory to virus infection. 26512077 6 126 theme carnivore 1322:1330 arg1 hosts 1332:1336 carnivore hosts 1322:1336 carnivore hosts used in the experimental evolution studies 1322:1379 Analysis of the TfRs of carnivore hosts used in the experimental evolution studies demonstrated that their glycosylation patterns varied, including a glycan present only on the domestic dog TfR that dictates susceptibility to parvoviruses. 26512077 1 127 theme important 212:220 arg1 carnivores 149:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores 129:158 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 127 theme important 212:220 arg1 raccoons 169:176 raccoons 169:176 raccoons 169:176 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 26512077 1 127 theme important 212:220 arg1 hosts 222:226 important hosts 212:226 important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs 212:308 UNLABELLED Sylvatic carnivores, such as raccoons, have recently been recognized as important hosts in the evolution of canine parvovirus (CPV), a pandemic pathogen of domestic dogs. 28421405 7 0 theme UVPD 1511:1514 arg1 yields 1529:1534 improved UVPD fragment ion yields 1502:1534 improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments 1502:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 4 1 theme yield 926:930 arg1 gains 932:936 fragment yield gains 917:936 fragment yield gains of an order of magnitude versus sodium adducts 917:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 4 2 theme Ion 766:768 arg1 mobility 770:777 Ion mobility and UVPD-MS spectra 766:797 mobility 770:777 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 8 3 theme Graphical 1625:1633 arg1 Abstract 1635:1642 Graphical Abstract 1625:1642 Graphical Abstract 1625:1642 Graphical Abstract ᅟ. 28421405 0 4 theme Mobility-Mass 98:110 arg1 Spectrometry 112:123 Ion Mobility-Mass Spectrometry 94:123 Ion Mobility-Mass Spectrometry 94:123 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 3 5 theme mobility 749:756 arg1 system 758:763 a drift tube ion mobility system 732:763 a drift tube ion mobility system 732:763 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 4 6 theme fragment 917:924 arg1 gains 932:936 fragment yield gains 917:936 fragment yield gains of an order of magnitude versus sodium adducts 917:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 2 7 theme UV 422:423 arg1 absorbance 425:434 strong UV absorbance 415:434 strong UV absorbance 415:434 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 1 8 theme biomolecule 309:319 arg1 identification 321:334 biomolecule identification 309:334 biomolecule identification 309:334 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 5 9 theme standards 1195:1203 arg1 isomers 1098:1104 the structural glycan isomers 1076:1104 the structural glycan isomers 1076:1104 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 9 theme standards 1195:1203 arg1 spectra 1175:1181 composite spectra 1165:1181 composite spectra of isomeric standards 1165:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 9 theme standards 1195:1203 arg1 mixture 1142:1148 a mixture 1140:1148 a mixture 1140:1148 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 6 10 theme discernable 1347:1357 arg1 separation 1359:1368 no discernable separation 1344:1368 no discernable separation obtained with the sodium adducts 1344:1401 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 5 11 theme glycan 1091:1096 arg1 mixture 1142:1148 a mixture 1140:1148 a mixture 1140:1148 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 11 theme glycan 1091:1096 arg1 spectra 1175:1181 composite spectra 1165:1181 composite spectra of isomeric standards 1165:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 11 theme glycan 1091:1096 arg1 isomers 1098:1104 the structural glycan isomers 1076:1104 the structural glycan isomers 1076:1104 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 3 12 theme adduction 605:613 arg1 capacity 587:594 the capacity 583:594 the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns 583:669 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 4 13 theme N-acetyl 807:814 arg1 isomers 823:829 two N-acetyl glycan isomers 803:829 two N-acetyl glycan isomers 803:829 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 7 14 theme distinction 1485:1495 arg1 It 1404:1405 It 1404:1405 It 1404:1405 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 14 theme distinction 1485:1495 arg1 enhancement 1423:1433 the combined enhancement 1410:1433 the combined enhancement of both isomeric drift time separation and isomer distinction 1410:1495 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 5 15 dep incorporated 1019:1030 arg1 identified 1121:1130 identified 1121:1130 could still be identified even as a mixture and not simply composite spectra of isomeric standards 1106:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 7 16 theme ion 1525:1527 arg1 yields 1529:1534 improved UVPD fragment ion yields 1502:1534 improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments 1502:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 4 17 theme UVPD-MS 783:789 arg1 spectra 791:797 Ion mobility and UVPD-MS spectra 766:797 spectra 791:797 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 3 18 theme glycans 564:570 arg1 modification 548:559 covalent modification 539:559 covalent modification of glycans 539:570 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 7 19 theme isomeric 1443:1450 arg1 separation 1463:1472 isomeric drift time separation 1443:1472 isomeric drift time separation 1443:1472 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 5 20 theme front-end 1032:1040 arg1 separation 1055:1064 front-end ion mobility separation 1032:1064 front-end ion mobility separation 1032:1064 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 1 21 theme collision-induced 228:244 arg1 CID 260:262 CID 260:262 CID 260:262 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 1 21 theme collision-induced 228:244 arg1 dissociation 246:257 collision-induced dissociation 228:257 collision-induced dissociation (CID) 228:263 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 5 22 dep identified 1121:1130 arg1 such 1066:1069 such 1066:1069 such 1066:1069 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 6 23 theme Cobalt 1206:1211 arg1 adduction 1213:1221 Cobalt adduction 1206:1221 Cobalt adduction 1206:1221 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 3 24 theme metal 599:603 arg1 adduction 605:613 metal adduction 599:613 metal adduction 599:613 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 3 25 theme UVPD 508:511 arg1 experiments 513:523 UVPD experiments 508:523 UVPD experiments that leverage covalent modification of glycans 508:570 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 7 26 theme time 1458:1461 arg1 separation 1463:1472 isomeric drift time separation 1443:1472 isomeric drift time separation 1443:1472 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 5 27 theme composite 1165:1173 arg1 spectra 1175:1181 composite spectra 1165:1181 composite spectra of isomeric standards 1165:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 27 theme composite 1165:1173 arg1 isomers 1098:1104 the structural glycan isomers 1076:1104 the structural glycan isomers 1076:1104 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 3 28 theme rich 638:641 arg1 patterns 662:669 comparatively rich UVPD fragmentation patterns 624:669 comparatively rich UVPD fragmentation patterns 624:669 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 2 29 theme low 450:452 arg1 yields 468:473 low fragmentation yields 450:473 low fragmentation yields for UVPD spectra 450:490 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 0 30 theme Differential 0:11 arg1 Fragmentation 13:25 Differential Fragmentation 0:25 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry 0:123 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 7 31 theme fragment 1516:1523 arg1 yields 1529:1534 improved UVPD fragment ion yields 1502:1534 improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments 1502:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 32 theme glycan 1599:1604 arg1 experiments 1612:1622 glycan IM-MS experiments 1599:1622 glycan IM-MS experiments 1599:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 3 33 theme UVPD 643:646 arg1 patterns 662:669 comparatively rich UVPD fragmentation patterns 624:669 comparatively rich UVPD fragmentation patterns 624:669 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 5 34 theme isomeric 1186:1193 arg1 standards 1195:1203 isomeric standards 1186:1203 isomeric standards 1186:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 0 35 theme Mobility-Selected 30:46 arg1 Glycans 48:54 Mobility-Selected Glycans 30:54 Mobility-Selected Glycans 30:54 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 4 36 theme sodium 865:870 arg1 cations 882:888 sodium or cobalt cations 865:888 sodium or cobalt cations 865:888 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 4 37 theme sodium 970:975 arg1 adducts 977:983 sodium adducts 970:983 sodium adducts 970:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 7 38 theme drift 1452:1456 arg1 separation 1463:1472 isomeric drift time separation 1443:1472 isomeric drift time separation 1443:1472 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 2 39 theme underivatized 365:377 arg1 glycans 379:385 underivatized glycans 365:385 underivatized glycans 365:385 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 6 40 theme 0.78 1303:1306 arg1 resolution 1289:1298 an isomer resolution 1279:1298 an isomer resolution of 0.78 when analyzed simultaneously 1279:1335 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 7 41 theme combined 1414:1421 arg1 It 1404:1405 It 1404:1405 It 1404:1405 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 41 theme combined 1414:1421 arg1 enhancement 1423:1433 the combined enhancement 1410:1433 the combined enhancement of both isomeric drift time separation and isomer distinction 1410:1495 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 42 theme IM-MS 1606:1610 arg1 experiments 1612:1622 glycan IM-MS experiments 1599:1622 glycan IM-MS experiments 1599:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 43 theme improved 1502:1509 arg1 yields 1529:1534 improved UVPD fragment ion yields 1502:1534 improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments 1502:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 44 with enhancement 1423:1433 arg1 yields 1529:1534 improved UVPD fragment ion yields 1502:1534 improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments 1502:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 1 45 theme dissociation 142:153 arg1 pathways 155:162 The alternative dissociation pathways 126:162 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) 126:263 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 3 46 theme covalent 539:546 arg1 modification 548:559 covalent modification 539:559 covalent modification of glycans 539:570 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 4 47 theme cobalt 875:880 arg1 cations 882:888 sodium or cobalt cations 865:888 sodium or cobalt cations 865:888 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 2 48 theme UVPD 479:482 arg1 spectra 484:490 UVPD spectra 479:490 UVPD spectra 479:490 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 0 49 theme Glycans 48:54 arg1 Fragmentation 13:25 Differential Fragmentation 0:25 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry 0:123 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 5 50 dep simply 1158:1163 arg1 not 1154:1156 not 1154:1156 not 1154:1156 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 4 51 theme adducts 977:983 arg1 order 944:948 an order 941:948 an order of magnitude versus sodium adducts 941:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 3 52 theme isomeric 709:716 arg1 set 725:727 an isomeric glycan set 706:727 an isomeric glycan set 706:727 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 5 53 theme structural 1080:1089 arg1 mixture 1142:1148 a mixture 1140:1148 a mixture 1140:1148 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 53 theme structural 1080:1089 arg1 spectra 1175:1181 composite spectra 1165:1181 composite spectra of isomeric standards 1165:1203 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 5 53 theme structural 1080:1089 arg1 isomers 1098:1104 the structural glycan isomers 1076:1104 the structural glycan isomers 1076:1104 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 7 54 theme multivalent 1557:1567 arg1 adduction 1575:1583 multivalent metal adduction 1557:1583 multivalent metal adduction for advancing glycan IM-MS experiments 1557:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 4 55 theme glycan 816:821 arg1 isomers 823:829 two N-acetyl glycan isomers 803:829 two N-acetyl glycan isomers 803:829 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 4 56 theme order 944:948 arg1 gains 932:936 fragment yield gains 917:936 fragment yield gains of an order of magnitude versus sodium adducts 917:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 1 57 dep useful 277:282 arg1 diagnostic 284:293 diagnostic 284:293 diagnostic 284:293 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 3 58 theme fragmentation 648:660 arg1 patterns 662:669 comparatively rich UVPD fragmentation patterns 624:669 comparatively rich UVPD fragmentation patterns 624:669 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 2 59 theme strong 415:420 arg1 absorbance 425:434 strong UV absorbance 415:434 strong UV absorbance 415:434 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 0 60 theme Ultraviolet 60:70 arg1 Photodissociation 72:88 Ultraviolet Photodissociation 60:88 Ultraviolet Photodissociation 60:88 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 6 61 theme sodium 1388:1393 arg1 adducts 1395:1401 the sodium adducts 1384:1401 the sodium adducts 1384:1401 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 3 62 theme glycan 718:723 arg1 set 725:727 an isomeric glycan set 706:727 an isomeric glycan set 706:727 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 6 63 theme isomer 1282:1287 arg1 resolution 1289:1298 an isomer resolution 1279:1298 an isomer resolution of 0.78 when analyzed simultaneously 1279:1335 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 4 64 theme magnitude 953:961 arg1 order 944:948 an order 941:948 an order of magnitude versus sodium adducts 941:983 Ion mobility and UVPD-MS spectra for two N-acetyl glycan isomers were examined, each adducted with sodium or cobalt cations, with the latter providing fragment yield gains of an order of magnitude versus sodium adducts. 28421405 3 65 theme ion 745:747 arg1 system 758:763 a drift tube ion mobility system 732:763 a drift tube ion mobility system 732:763 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 3 66 dep experiments 513:523 arg1 contrast 496:503 contrast 496:503 contrast 496:503 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 7 67 theme separation 1463:1472 arg1 It 1404:1405 It 1404:1405 It 1404:1405 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 67 theme separation 1463:1472 arg1 enhancement 1423:1433 the combined enhancement 1410:1433 the combined enhancement of both isomeric drift time separation and isomer distinction 1410:1495 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 7 68 theme metal 1569:1573 arg1 adduction 1575:1583 multivalent metal adduction 1557:1583 multivalent metal adduction for advancing glycan IM-MS experiments 1557:1622 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 1 69 theme alternative 130:140 arg1 pathways 155:162 The alternative dissociation pathways 126:162 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) 126:263 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 7 70 theme isomer 1478:1483 arg1 distinction 1485:1495 isomer distinction 1478:1495 isomer distinction 1478:1495 It is the combined enhancement of both isomeric drift time separation and isomer distinction with improved UVPD fragment ion yields that further bolster multivalent metal adduction for advancing glycan IM-MS experiments. 28421405 5 71 theme ion 1042:1044 arg1 separation 1055:1064 front-end ion mobility separation 1032:1064 front-end ion mobility separation 1032:1064 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 6 72 theme glycan 1249:1254 arg1 separation 1256:1265 the glycan separation 1245:1265 the glycan separation 1245:1265 Cobalt adduction proved influential in the glycan separation by yielding an isomer resolution of 0.78 when analyzed simultaneously versus no discernable separation obtained with the sodium adducts. 28421405 1 73 theme ultraviolet 177:187 arg1 UVPD 208:211 UVPD 208:211 UVPD 208:211 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 1 73 theme ultraviolet 177:187 arg1 photodissociation 189:205 ultraviolet photodissociation 177:205 ultraviolet photodissociation (UVPD) 177:212 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 1 74 theme useful 277:282 arg1 fragments 295:303 useful diagnostic fragments 277:303 useful diagnostic fragments 277:303 The alternative dissociation pathways initiated by ultraviolet photodissociation (UVPD) compared with collision-induced dissociation (CID) may provide useful diagnostic fragments for biomolecule identification, including glycans. 28421405 0 75 theme Ion 94:96 arg1 Spectrometry 112:123 Ion Mobility-Mass Spectrometry 94:123 Ion Mobility-Mass Spectrometry 94:123 Differential Fragmentation of Mobility-Selected Glycans via Ultraviolet Photodissociation and Ion Mobility-Mass Spectrometry. 28421405 3 76 theme drift 734:738 arg1 system 758:763 a drift tube ion mobility system 732:763 a drift tube ion mobility system 732:763 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 3 77 theme separation 683:692 arg1 factors 694:700 separation factors 683:700 separation factors for an isomeric glycan set 683:727 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 5 78 theme glycan 1003:1008 arg1 analysis 1010:1017 our glycan analysis 999:1017 our glycan analysis 999:1017 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 28421405 2 79 theme fragmentation 454:466 arg1 yields 468:473 low fragmentation yields 450:473 low fragmentation yields for UVPD spectra 450:490 However, underivatized glycans do not commonly demonstrate strong UV absorbance, resulting in low fragmentation yields for UVPD spectra. 28421405 3 80 theme tube 740:743 arg1 system 758:763 a drift tube ion mobility system 732:763 a drift tube ion mobility system 732:763 In contrast to UVPD experiments that leverage covalent modification of glycans, we detail the capacity of metal adduction to yield comparatively rich UVPD fragmentation patterns and enhance separation factors for an isomeric glycan set in a drift tube ion mobility system. 28421405 5 81 theme mobility 1046:1053 arg1 separation 1055:1064 front-end ion mobility separation 1032:1064 front-end ion mobility separation 1032:1064 Furthermore, our glycan analysis incorporated front-end ion mobility separation such that the structural glycan isomers could still be identified even as a mixture and not simply composite spectra of isomeric standards. 26894673 11 0 theme preferred 1782:1790 arg1 conformation 1819:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 2 1 theme cellulosic 133:142 arg1 biomass 150:156 cellulosic plant biomass 133:156 cellulosic plant biomass 133:156 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 7 2 theme sequence 974:981 arg1 identity 983:990 78% sequence identity 970:990 78% sequence identity 970:990 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 3 theme 78 970:971 arg1 % 972:972 % 972:972 % 972:972 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 12 4 theme variant 2151:2157 arg1 enzymes 2159:2165 variant enzymes 2151:2165 variant enzymes 2151:2165 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 4 5 theme major 458:462 arg1 drawback 464:471 A major drawback 456:471 A major drawback with this process 456:489 A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. 26894673 6 6 from fumigatus 839:847 arg1 structures 753:762 the three-dimensional structures 731:762 the three-dimensional structures 731:762 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 6 6 from fumigatus 839:847 arg1 β-D-glucosidases 805:820 two industrially important family GH3 β-D-glucosidases 767:820 two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae 767:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 2 7 theme useful 163:168 arg1 products 170:177 useful products 163:177 useful products such as biofuels 163:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 2 7 theme useful 163:168 arg1 biofuels 187:194 biofuels 187:194 biofuels 187:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 6 8 theme 1.95 Å 911:916 arg1 resolution 918:927 1.95 Å resolution 911:927 1.95 Å resolution 911:927 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 7 9 dep family 1196:1201 arg1 GH3 1203:1205 GH3 1203:1205 glycoside hydrolase family GH3 1176:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 3 10 theme plant 257:261 arg1 enzymes 273:279 diverse plant degrading enzymes 249:279 diverse plant degrading enzymes 249:279 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 8 11 theme GlcNAc 1312:1317 arg1 molecule 1319:1326 a single GlcNAc molecule 1303:1326 a single GlcNAc molecule 1303:1326 Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. 26894673 9 12 theme N-linked 1343:1350 arg1 bonds 1458:1462 multiple hydrogen bonds 1440:1462 multiple hydrogen bonds between their sugar components and adjacent protein side chains 1440:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 12 theme N-linked 1343:1350 arg1 trees 1419:1423 high-mannose trees 1406:1423 high-mannose trees 1406:1423 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 12 theme N-linked 1343:1350 arg1 glycans 1335:1341 Those glycans 1329:1341 Those glycans N-linked to the core of the structure 1329:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 3 13 theme classical 282:290 arg1 cellulases 313:322 classical exo- and endo-acting cellulases 282:322 classical exo- and endo-acting cellulases 282:322 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 6 14 from structures 753:762 arg1 oryzae 856:861 A. oryzae 853:861 A. oryzae 853:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 6 14 from structures 753:762 arg1 fumigatus 839:847 Aspergillus fumigatus 827:847 Aspergillus fumigatus 827:847 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 7 15 theme glycoside 1176:1184 arg1 family 1196:1201 glycoside hydrolase family 1176:1201 glycoside hydrolase family GH3 1176:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 4 16 theme exo-acting 503:512 arg1 cellobiohydrolases 514:531 the exo-acting cellobiohydrolases 499:531 the exo-acting cellobiohydrolases 499:531 A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. 26894673 7 17 theme β-D-glucan 1101:1110 arg1 structure 1154:1162 the first three-dimensional structure 1126:1162 the first three-dimensional structure solved from glycoside hydrolase family GH3 1126:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 17 theme β-D-glucan 1101:1110 arg1 exohydrolase 1112:1123 barley β-D-glucan exohydrolase 1094:1123 barley β-D-glucan exohydrolase 1094:1123 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 18 theme three-dimensional 1136:1152 arg1 structure 1154:1162 the first three-dimensional structure 1126:1162 the first three-dimensional structure solved from glycoside hydrolase family GH3 1126:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 18 theme three-dimensional 1136:1152 arg1 exohydrolase 1112:1123 barley β-D-glucan exohydrolase 1094:1123 barley β-D-glucan exohydrolase 1094:1123 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 9 19 theme structure 1371:1379 arg1 core 1359:1362 the core 1355:1362 the core of the structure 1355:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 3 20 theme lytic 360:364 arg1 monooxygenases 381:394 cellulose-active lytic polysaccharide monooxygenases 343:394 cellulose-active lytic polysaccharide monooxygenases 343:394 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 6 21 theme three-dimensional 735:751 arg1 structures 753:762 the three-dimensional structures 731:762 the three-dimensional structures 731:762 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 12 22 theme new 2132:2134 arg1 generations 2136:2146 new generations 2132:2146 new generations of variant enzymes for industrial application 2132:2192 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 11 23 theme structures 1901:1910 arg1 refinements 1872:1882 refinements 1872:1882 refinements of other crystal structures containing O- or N-glycosylation 1872:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 12 24 theme fungal 2054:2059 arg1 β-D-glucosidases 2061:2076 fungal β-D-glucosidases 2054:2076 fungal β-D-glucosidases 2054:2076 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 9 25 link N-linked 1343:1350 arg1 bonds 1458:1462 multiple hydrogen bonds 1440:1462 multiple hydrogen bonds between their sugar components and adjacent protein side chains 1440:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 25 link N-linked 1343:1350 arg1 trees 1419:1423 high-mannose trees 1406:1423 high-mannose trees 1406:1423 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 25 link N-linked 1343:1350 arg1 glycans 1335:1341 Those glycans 1329:1341 Those glycans N-linked to the core of the structure 1329:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 4 26 theme cellobiose 574:583 arg1 product 585:591 their cellobiose product 568:591 their cellobiose product 568:591 A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. 26894673 10 27 theme special 1556:1562 arg1 problems 1564:1571 special problems 1556:1571 special problems for crystallographic refinement 1556:1603 The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. 26894673 11 28 theme other 1887:1891 arg1 structures 1901:1910 other crystal structures 1887:1910 other crystal structures containing O- or N-glycosylation 1887:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 5 29 theme product 705:711 arg1 inhibition 713:722 limiting product inhibition 696:722 limiting product inhibition 696:722 β-D-Glucosidases are therefore important for liberating glucose from cellobiose and thereby relieving limiting product inhibition. 26894673 1 30 theme family 75:80 arg1 β-D-glucosidases 86:101 family GH3 β-D-glucosidases 75:101 family GH3 β-D-glucosidases. 75:102 family GH3 β-D-glucosidases. 26894673 3 31 theme β-D-linked 429:438 arg1 polysaccharide 440:453 the recalcitrant β-D-linked polysaccharide 412:453 the recalcitrant β-D-linked polysaccharide 412:453 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 4 32 theme severe 545:550 arg1 inhibition 552:561 severe inhibition 545:561 severe inhibition 545:561 A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. 26894673 6 33 theme β-D-glucosidases 805:820 arg1 structures 753:762 the three-dimensional structures 731:762 the three-dimensional structures 731:762 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 0 34 theme Three-dimensional 0:16 arg1 structures 18:27 Three-dimensional structures 0:27 Three-dimensional structures of two 0:34 Three-dimensional structures of two heavily N-glycosylated Aspergillus sp. 26894673 9 35 theme multiple 1440:1447 arg1 bonds 1458:1462 multiple hydrogen bonds 1440:1462 multiple hydrogen bonds between their sugar components and adjacent protein side chains 1440:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 35 theme multiple 1440:1447 arg1 trees 1419:1423 high-mannose trees 1406:1423 high-mannose trees 1406:1423 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 35 theme multiple 1440:1447 arg1 glycans 1335:1341 Those glycans 1329:1341 Those glycans N-linked to the core of the structure 1329:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 6 36 theme family 794:799 arg1 β-D-glucosidases 805:820 two industrially important family GH3 β-D-glucosidases 767:820 two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae 767:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 12 37 theme other 1990:1994 arg1 structures 2021:2030 other recent three-dimensional structures 1990:2030 other recent three-dimensional structures 1990:2030 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 12 38 theme three-dimensional 2003:2019 arg1 structures 2021:2030 other recent three-dimensional structures 1990:2030 other recent three-dimensional structures 1990:2030 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 12 39 theme Aspergillus 1950:1960 arg1 structures 1966:1975 The Aspergillus GH3 structures 1946:1975 The Aspergillus GH3 structures 1946:1975 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 11 40 theme chair 1813:1817 arg1 conformation 1819:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 6 41 theme molecular 874:882 arg1 replacement 884:894 molecular replacement 874:894 molecular replacement 874:894 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 0 42 theme Aspergillus 59:69 arg1 sp 71:72 N-glycosylated Aspergillus sp 44:72 N-glycosylated Aspergillus sp 44:72 Three-dimensional structures of two heavily N-glycosylated Aspergillus sp. 26894673 11 43 theme general 1849:1855 arg1 application 1857:1867 general application 1849:1867 general application to refinements of other crystal structures containing O- or N-glycosylation 1849:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 9 44 theme adjacent 1499:1506 arg1 chains 1521:1526 adjacent protein side chains 1499:1526 adjacent protein side chains 1499:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 0 45 gly N-glycosylated 44:57 arg1 sp 71:72 N-glycosylated Aspergillus sp 44:72 N-glycosylated Aspergillus sp 44:72 Three-dimensional structures of two heavily N-glycosylated Aspergillus sp. 26894673 2 46 theme major 201:205 arg1 goal 216:219 a major societal goal 199:219 a major societal goal 199:219 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 2 46 theme major 201:205 arg1 conversion 119:128 The industrial conversion 104:128 The industrial conversion of cellulosic plant biomass into useful products such as biofuels 104:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 7 47 theme three-domain 1003:1014 arg1 structure 1016:1024 a three-domain structure 1001:1024 a three-domain structure with the catalytic domain at the interface 1001:1067 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 9 48 theme side 1516:1519 arg1 chains 1521:1526 adjacent protein side chains 1499:1526 adjacent protein side chains 1499:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 12 49 theme enzymes 2159:2165 arg1 generations 2136:2146 new generations 2132:2146 new generations of variant enzymes for industrial application 2132:2192 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 2 50 theme plant 144:148 arg1 biomass 150:156 cellulosic plant biomass 133:156 cellulosic plant biomass 133:156 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 11 51 theme glycosylation 1741:1753 arg1 trees 1755:1759 the glycosylation trees 1737:1759 the glycosylation trees 1737:1759 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 12 52 theme industrial 2171:2180 arg1 application 2182:2192 industrial application 2171:2192 industrial application 2171:2192 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 3 53 theme diverse 249:255 arg1 enzymes 273:279 diverse plant degrading enzymes 249:279 diverse plant degrading enzymes 249:279 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 6 54 from oryzae 856:861 arg1 structures 753:762 the three-dimensional structures 731:762 the three-dimensional structures 731:762 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 6 54 from oryzae 856:861 arg1 β-D-glucosidases 805:820 two industrially important family GH3 β-D-glucosidases 767:820 two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae 767:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 8 55 theme external 1269:1276 arg1 sites 1278:1282 only a few external sites 1258:1282 only a few external sites 1258:1282 Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. 26894673 11 56 contain containing 1912:1921 arg2 O- 1923:1924 O- 1923:1924 O- 1923:1924 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 11 56 contain containing 1912:1921 arg1 structures 1901:1910 other crystal structures 1887:1910 other crystal structures containing O- or N-glycosylation 1887:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 11 56 contain containing 1912:1921 arg2 N-glycosylation 1929:1943 N-glycosylation 1929:1943 N-glycosylation 1929:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 7 57 theme % 972:972 arg1 identity 983:990 78% sequence identity 970:990 78% sequence identity 970:990 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 58 with structure 1016:1024 arg1 domain 1045:1050 the catalytic domain 1031:1050 the catalytic domain at the interface 1031:1067 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 10 59 theme extensive 1533:1541 arg1 glycans 1543:1549 The extensive glycans 1529:1549 The extensive glycans 1529:1549 The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. 26894673 3 60 theme degrading 263:271 arg1 enzymes 273:279 diverse plant degrading enzymes 249:279 diverse plant degrading enzymes 249:279 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 7 61 theme hydrolase 1186:1194 arg1 family 1196:1201 glycoside hydrolase family 1176:1201 glycoside hydrolase family GH3 1176:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 8 62 theme single 1305:1310 arg1 molecule 1319:1326 a single GlcNAc molecule 1303:1326 a single GlcNAc molecule 1303:1326 Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. 26894673 3 63 theme endo-acting 301:311 arg1 cellulases 313:322 classical exo- and endo-acting cellulases 282:322 classical exo- and endo-acting cellulases 282:322 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 11 64 from D-pyranosides 1720:1732 arg1 trees 1755:1759 the glycosylation trees 1737:1759 the glycosylation trees 1737:1759 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 3 65 theme exo- 292:295 arg1 cellulases 313:322 classical exo- and endo-acting cellulases 282:322 classical exo- and endo-acting cellulases 282:322 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 2 66 theme industrial 108:117 arg1 goal 216:219 a major societal goal 199:219 a major societal goal 199:219 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 2 66 theme industrial 108:117 arg1 conversion 119:128 The industrial conversion 104:128 The industrial conversion of cellulosic plant biomass into useful products such as biofuels 104:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 7 67 theme first 1130:1134 arg1 structure 1154:1162 the first three-dimensional structure 1126:1162 the first three-dimensional structure solved from glycoside hydrolase family GH3 1126:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 67 theme first 1130:1134 arg1 exohydrolase 1112:1123 barley β-D-glucan exohydrolase 1094:1123 barley β-D-glucan exohydrolase 1094:1123 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 6 68 dep structures 753:762 arg1 refined 900:906 refined 900:906 refined at 1.95 Å resolution 900:927 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 6 68 dep structures 753:762 arg1 solved 864:869 solved 864:869 solved by molecular replacement 864:894 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 3 69 link β-D-linked 429:438 arg1 polysaccharide 440:453 the recalcitrant β-D-linked polysaccharide 412:453 the recalcitrant β-D-linked polysaccharide 412:453 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 7 70 theme barley 1094:1099 arg1 structure 1154:1162 the first three-dimensional structure 1126:1162 the first three-dimensional structure solved from glycoside hydrolase family GH3 1126:1205 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 7 70 theme barley 1094:1099 arg1 exohydrolase 1112:1123 barley β-D-glucan exohydrolase 1094:1123 barley β-D-glucan exohydrolase 1094:1123 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 3 71 theme polysaccharide 366:379 arg1 monooxygenases 381:394 cellulose-active lytic polysaccharide monooxygenases 343:394 cellulose-active lytic polysaccharide monooxygenases 343:394 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 7 72 from interface 1059:1067 arg1 domain 1045:1050 the catalytic domain 1031:1050 the catalytic domain at the interface 1031:1067 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 3 73 theme cellulose-active 343:358 arg1 monooxygenases 381:394 cellulose-active lytic polysaccharide monooxygenases 343:394 cellulose-active lytic polysaccharide monooxygenases 343:394 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 9 74 theme high-mannose 1406:1417 arg1 bonds 1458:1462 multiple hydrogen bonds 1440:1462 multiple hydrogen bonds between their sugar components and adjacent protein side chains 1440:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 74 theme high-mannose 1406:1417 arg1 trees 1419:1423 high-mannose trees 1406:1423 high-mannose trees 1406:1423 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 74 theme high-mannose 1406:1417 arg1 glycans 1335:1341 Those glycans 1329:1341 Those glycans N-linked to the core of the structure 1329:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 5 75 theme limiting 696:703 arg1 inhibition 713:722 limiting product inhibition 696:722 limiting product inhibition 696:722 β-D-Glucosidases are therefore important for liberating glucose from cellobiose and thereby relieving limiting product inhibition. 26894673 11 76 theme crystal 1893:1899 arg1 structures 1901:1910 other crystal structures 1887:1910 other crystal structures containing O- or N-glycosylation 1887:1943 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 11 77 theme minimum-energy 1792:1805 arg1 conformation 1819:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 1 78 theme GH3 82:84 arg1 β-D-glucosidases 86:101 family GH3 β-D-glucosidases 75:101 family GH3 β-D-glucosidases. 75:102 family GH3 β-D-glucosidases. 26894673 10 79 theme crystallographic 1577:1592 arg1 refinement 1594:1603 crystallographic refinement 1577:1603 crystallographic refinement 1577:1603 The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. 26894673 6 80 theme GH3 801:803 arg1 β-D-glucosidases 805:820 two industrially important family GH3 β-D-glucosidases 767:820 two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae 767:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 0 81 theme two 32:34 arg1 structures 18:27 Three-dimensional structures 0:27 Three-dimensional structures of two 0:34 Three-dimensional structures of two heavily N-glycosylated Aspergillus sp. 26894673 9 82 theme hydrogen 1449:1456 arg1 bonds 1458:1462 multiple hydrogen bonds 1440:1462 multiple hydrogen bonds between their sugar components and adjacent protein side chains 1440:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 82 theme hydrogen 1449:1456 arg1 trees 1419:1423 high-mannose trees 1406:1423 high-mannose trees 1406:1423 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 9 82 theme hydrogen 1449:1456 arg1 glycans 1335:1341 Those glycans 1329:1341 Those glycans N-linked to the core of the structure 1329:1379 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 12 83 theme recent 1996:2001 arg1 structures 2021:2030 other recent three-dimensional structures 1990:2030 other recent three-dimensional structures 1990:2030 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 7 84 theme catalytic 1035:1043 arg1 domain 1045:1050 the catalytic domain 1031:1050 the catalytic domain at the interface 1031:1067 Both enzymes, which share 78% sequence identity, display a three-domain structure with the catalytic domain at the interface, as originally shown for barley β-D-glucan exohydrolase, the first three-dimensional structure solved from glycoside hydrolase family GH3. 26894673 3 85 theme recalcitrant 416:427 arg1 polysaccharide 440:453 the recalcitrant β-D-linked polysaccharide 412:453 the recalcitrant β-D-linked polysaccharide 412:453 These technologies harness diverse plant degrading enzymes, classical exo- and endo-acting cellulases and, increasingly, cellulose-active lytic polysaccharide monooxygenases, to deconstruct the recalcitrant β-D-linked polysaccharide. 26894673 6 86 theme important 784:792 arg1 β-D-glucosidases 805:820 two industrially important family GH3 β-D-glucosidases 767:820 two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae 767:861 Here, the three-dimensional structures of two industrially important family GH3 β-D-glucosidases from Aspergillus fumigatus and A. oryzae, solved by molecular replacement and refined at 1.95 Å resolution, are reported. 26894673 0 87 theme N-glycosylated 44:57 arg1 sp 71:72 N-glycosylated Aspergillus sp 44:72 N-glycosylated Aspergillus sp 44:72 Three-dimensional structures of two heavily N-glycosylated Aspergillus sp. 26894673 12 88 theme structures 2021:2030 arg1 light 1981:1985 light 1981:1985 light of other recent three-dimensional structures 1981:2030 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 10 89 theme new 1610:1612 arg1 techniques 1614:1623 new techniques 1610:1623 new techniques 1610:1623 The extensive glycans pose special problems for crystallographic refinement, and new techniques and protocols were developed especially for this work. 26894673 11 90 theme C1 1810:1811 arg1 conformation 1819:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 the preferred minimum-energy (4)C1 chair conformation 1778:1830 These protocols ensured that all of the D-pyranosides in the glycosylation trees were modelled in the preferred minimum-energy (4)C1 chair conformation and should be of general application to refinements of other crystal structures containing O- or N-glycosylation. 26894673 9 91 theme sugar 1478:1482 arg1 components 1484:1493 their sugar components 1472:1493 their sugar components 1472:1493 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 8 92 theme extensive 1226:1234 arg1 N-glycosylation 1236:1250 extensive N-glycosylation 1226:1250 extensive N-glycosylation 1226:1250 Both enzymes show extensive N-glycosylation, with only a few external sites being truncated to a single GlcNAc molecule. 26894673 12 93 from structures 1966:1975 arg1 light 1981:1985 light 1981:1985 light of other recent three-dimensional structures 1981:2030 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 12 94 theme GH3 1962:1964 arg1 structures 1966:1975 The Aspergillus GH3 structures 1946:1975 The Aspergillus GH3 structures 1946:1975 The Aspergillus GH3 structures, in light of other recent three-dimensional structures, provide insight into fungal β-D-glucosidases and provide a platform on which to inform and inspire new generations of variant enzymes for industrial application. 26894673 2 95 theme societal 207:214 arg1 goal 216:219 a major societal goal 199:219 a major societal goal 199:219 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 2 95 theme societal 207:214 arg1 conversion 119:128 The industrial conversion 104:128 The industrial conversion of cellulosic plant biomass into useful products such as biofuels 104:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 9 96 theme protein 1508:1514 arg1 chains 1521:1526 adjacent protein side chains 1499:1526 adjacent protein side chains 1499:1526 Those glycans N-linked to the core of the structure are identified purely as high-mannose trees, and establish multiple hydrogen bonds between their sugar components and adjacent protein side chains. 26894673 2 97 theme biomass 150:156 arg1 goal 216:219 a major societal goal 199:219 a major societal goal 199:219 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 2 97 theme biomass 150:156 arg1 conversion 119:128 The industrial conversion 104:128 The industrial conversion of cellulosic plant biomass into useful products such as biofuels 104:194 The industrial conversion of cellulosic plant biomass into useful products such as biofuels is a major societal goal. 26894673 4 98 with drawback 464:471 arg1 process 483:489 this process 478:489 this process 478:489 A major drawback with this process is that the exo-acting cellobiohydrolases suffer from severe inhibition from their cellobiose product. 25676505 7 0 gly glycosylated 935:946 arg1 product 948:954 glycosylated product 935:954 glycosylated product 935:954 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 6 1 theme other 826:830 arg1 hand 832:835 the other hand 822:835 the other hand 822:835 On the other hand, GlcNAc-2C12 primer was not elongated. 25676505 7 2 dep C6 1035:1036 arg1 C12 1026:1028 C12 1026:1028 C12 1026:1028 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 2 3 with glycosylation 449:461 arg1 peracetylglucosamine 536:555 peracetylglucosamine 536:555 peracetylglucosamine 536:555 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 7 4 theme same 1064:1067 arg1 number 1069:1074 almost the same number 1053:1074 almost the same number of C-units 1053:1085 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 0 5 from Effect 0:5 arg1 elongation 42:51 saccharide elongation 31:51 saccharide elongation by cells 31:60 Effect of aglycon structure on saccharide elongation by cells. 25676505 1 6 used used 166:169 arg2 primers 104:110 GlcNAc primers 97:110 GlcNAc primers 97:110 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 1 6 used used 166:169 arg2 N-acetyl-β-D-glucosaminide 69:94 Alkyl N-acetyl-β-D-glucosaminide 63:94 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties 63:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 7 7 theme aglycon 993:999 arg1 structure 1001:1009 the aglycon structure 989:1009 the aglycon structure of GlcNAc 989:1019 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 1 8 theme Alkyl 63:67 arg1 N-acetyl-β-D-glucosaminide 69:94 Alkyl N-acetyl-β-D-glucosaminide 63:94 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties 63:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 1 8 theme Alkyl 63:67 arg1 primers 104:110 GlcNAc primers 97:110 GlcNAc primers 97:110 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 2 9 gly glycosylation 449:461 arg1 dodecan-1-ol 466:477 dodecan-1-ol 466:477 dodecan-1-ol 466:477 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 2 10 theme tridecan-7-yl 304:316 arg1 GlcNAc-2C6 346:355 GlcNAc-2C6 346:355 GlcNAc-2C6 346:355 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 2 10 theme tridecan-7-yl 304:316 arg1 N-acetyl-β-D-glucosaminide 318:343 tridecan-7-yl N-acetyl-β-D-glucosaminide 304:343 tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6) 304:356 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 1 11 theme GlcNAc 97:102 arg1 N-acetyl-β-D-glucosaminide 69:94 Alkyl N-acetyl-β-D-glucosaminide 63:94 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties 63:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 1 11 theme GlcNAc 97:102 arg1 primers 104:110 GlcNAc primers 97:110 GlcNAc primers 97:110 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 7 12 theme glycosylated 935:946 arg1 product 948:954 glycosylated product 935:954 glycosylated product 935:954 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 0 13 theme structure 18:26 arg1 Effect 0:5 Effect 0:5 Effect of aglycon structure on saccharide elongation by cells 0:60 Effect of aglycon structure on saccharide elongation by cells. 25676505 7 14 theme product 948:954 arg1 amount 925:930 the amount 921:930 the amount of glycosylated product 921:954 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 7 14 theme product 948:954 arg1 product 948:954 glycosylated product 935:954 glycosylated product 935:954 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 0 15 theme aglycon 10:16 arg1 structure 18:26 aglycon structure 10:26 aglycon structure 10:26 Effect of aglycon structure on saccharide elongation by cells. 25676505 4 16 theme 48 h 646:649 arg1 incubation 651:660 48 h incubation 646:660 48 h incubation 646:660 After 48 h incubation, results showed that GlcNAc-C12 was elongated to give NeuAc-Gal-GlcNAc-C12. 25676505 3 17 theme melanoma 601:608 arg1 cells 610:614 mouse B16 melanoma cells 591:614 mouse B16 melanoma cells 591:614 These primers were introduced to mouse B16 melanoma cells to prepare glycolipids. 25676505 2 18 theme N-acetyl-β-D-glucosaminide 318:343 arg1 primers 421:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 0 19 theme saccharide 31:40 arg1 elongation 42:51 saccharide elongation 31:51 saccharide elongation by cells 31:60 Effect of aglycon structure on saccharide elongation by cells. 25676505 2 20 theme pentacosan-13-yl 363:378 arg1 GlcNAc-2C12 408:418 GlcNAc-2C12 408:418 GlcNAc-2C12 408:418 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 2 20 theme pentacosan-13-yl 363:378 arg1 N-acetyl-β-D-glucosaminide 380:405 pentacosan-13-yl N-acetyl-β-D-glucosaminide 363:405 pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) 363:419 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 1 21 theme aglycon 202:208 arg1 structure 210:218 the aglycon structure 198:218 the aglycon structure 198:218 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 2 22 theme dodecan-1-ol 466:477 arg1 glycosylation 449:461 glycosylation 449:461 glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine 449:555 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 1 23 with N-acetyl-β-D-glucosaminide 69:94 arg1 moieties 136:143 different aglycon moieties 118:143 different aglycon moieties 118:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 1 24 theme structure 210:218 arg1 effect 188:193 the effect 184:193 the effect of the aglycon structure on cellular saccharide elongation 184:252 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 2 25 theme N-acetyl-β-D-glucosaminide 380:405 arg1 primers 421:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 7 26 theme C-units 1079:1085 arg1 number 1069:1074 almost the same number 1053:1074 almost the same number of C-units 1053:1085 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 1 27 theme different 118:126 arg1 moieties 136:143 different aglycon moieties 118:143 different aglycon moieties 118:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 1 28 theme aglycon 128:134 arg1 moieties 136:143 different aglycon moieties 118:143 different aglycon moieties 118:143 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 2 29 theme N-acetyl-β-D-glucosaminide 263:288 arg1 primers 421:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers 255:427 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 6 30 theme GlcNAc-2C12 838:848 arg1 primer 850:855 GlcNAc-2C12 primer 838:855 GlcNAc-2C12 primer 838:855 On the other hand, GlcNAc-2C12 primer was not elongated. 25676505 7 31 theme GlcNAc 1014:1019 arg1 structure 1001:1009 the aglycon structure 989:1009 the aglycon structure of GlcNAc 989:1019 Significantly, the results demonstrated that the amount of glycosylated product increased 1.5-times by modifying the aglycon structure of GlcNAc from C12 to 2 C6 despite having almost the same number of C-units. 25676505 1 32 theme cellular 223:230 arg1 elongation 243:252 cellular saccharide elongation 223:252 cellular saccharide elongation 223:252 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 2 33 theme Dodecyl 255:261 arg1 GlcNAc-C12 291:300 GlcNAc-C12 291:300 GlcNAc-C12 291:300 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 2 33 theme Dodecyl 255:261 arg1 N-acetyl-β-D-glucosaminide 263:288 Dodecyl N-acetyl-β-D-glucosaminide 255:288 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12) 255:301 Dodecyl N-acetyl-β-D-glucosaminide (GlcNAc-C12), tridecan-7-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C6), and pentacosan-13-yl N-acetyl-β-D-glucosaminide (GlcNAc-2C12) primers were synthesized by glycosylation of dodecan-1-ol, tridecan-7-ol, and pentacosan-13-ol, respectively, with peracetylglucosamine. 25676505 1 34 from effect 188:193 arg1 elongation 243:252 cellular saccharide elongation 223:252 cellular saccharide elongation 223:252 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 3 35 theme mouse 591:595 arg1 cells 610:614 mouse B16 melanoma cells 591:614 mouse B16 melanoma cells 591:614 These primers were introduced to mouse B16 melanoma cells to prepare glycolipids. 25676505 1 36 theme saccharide 232:241 arg1 elongation 243:252 cellular saccharide elongation 223:252 cellular saccharide elongation 223:252 Alkyl N-acetyl-β-D-glucosaminide (GlcNAc primers) with different aglycon moieties were synthesized and used to determine the effect of the aglycon structure on cellular saccharide elongation. 25676505 3 37 theme B16 597:599 arg1 cells 610:614 mouse B16 melanoma cells 591:614 mouse B16 melanoma cells 591:614 These primers were introduced to mouse B16 melanoma cells to prepare glycolipids. 25434632 1 0 theme retrospective 168:180 arg1 analyses 213:220 retrospective in situ mass spectrometry (MS) analyses 168:220 retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples 168:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 5 1 dep imaging 1193:1199 arg1 iii 1182:1184 iii 1182:1184 iii 1182:1184 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 6 2 theme existing 1419:1426 arg1 strategies 1428:1437 existing strategies 1419:1437 existing strategies 1419:1437 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 6 3 theme N-glycans 1393:1401 arg1 characterization 1357:1372 characterization 1357:1372 characterization 1357:1372 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 6 3 theme N-glycans 1393:1401 arg1 imaging 1345:1351 MALDI imaging 1339:1351 MALDI imaging 1339:1351 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 1 4 theme in 182:183 arg1 spectrometry 195:206 in situ mass spectrometry 182:206 retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples 168:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 1 4 theme in 182:183 arg1 MS 209:210 MS 209:210 MS 209:210 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 5 5 theme liquid 1008:1013 arg1 chromatography 1015:1028 liquid chromatography 1008:1028 liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging 1008:1199 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 6 6 dep complete 1292:1299 arg1 straightforward 1302:1316 straightforward 1302:1316 straightforward 1302:1316 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 4 7 theme PNGase 673:678 arg1 release 691:697 in situ PNGase F-mediated release 665:697 in situ PNGase F-mediated release 665:697 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 8 theme data 807:810 arg1 sum 786:788 The sum 782:788 The sum of the presented data 782:810 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 9 theme composition 956:966 arg1 analysis 968:975 composition analysis 956:975 composition analysis 956:975 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 9 theme composition 956:966 arg1 strategies 925:934 three strategies 919:934 three strategies: (i) extraction and composition analysis 919:975 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 10 from arrays 1161:1166 arg1 situ 1171:1174 situ 1171:1174 situ 1171:1174 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 11 theme N-glycans 1239:1247 arg1 specificity 1224:1234 the tissue specificity 1213:1234 the tissue specificity of N-glycans at a higher resolution 1213:1270 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 2 12 theme laser 348:352 arg1 MALDI 377:381 MALDI 377:381 MALDI 377:381 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 2 12 theme laser 348:352 arg1 desorption/ionization 354:374 matrix-assisted laser desorption/ionization 332:374 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 5 13 theme droplet 1153:1159 arg1 arrays 1161:1166 large droplet arrays 1147:1166 large droplet arrays in situ 1147:1174 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 14 theme ionization 1054:1063 arg1 MS 1074:1075 electrospray ionization ion trap MS 1041:1075 electrospray ionization ion trap MS 1041:1075 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 4 15 theme kidney 774:779 arg1 sections 740:747 sections 740:747 sections of formalin-fixed murine kidney 740:779 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 4 16 from sections 740:747 arg1 measurement 703:713 measurement 703:713 measurement 703:713 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 4 16 from sections 740:747 arg1 release 691:697 in situ PNGase F-mediated release 665:697 in situ PNGase F-mediated release 665:697 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 4 17 theme murine 767:772 arg1 kidney 774:779 formalin-fixed murine kidney 752:779 formalin-fixed murine kidney 752:779 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 18 theme formalin-fixed 873:886 arg1 tissue 888:893 formalin-fixed tissue 873:893 formalin-fixed tissue 873:893 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 19 theme ion 1065:1067 arg1 MS 1074:1075 electrospray ionization ion trap MS 1041:1075 electrospray ionization ion trap MS 1041:1075 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 3 20 theme in 483:484 arg1 analyses 491:498 in situ analyses 483:498 in situ analyses of tissue-specific N-glycosylation profiles 483:542 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 4 21 theme glycans 727:733 arg1 measurement 703:713 measurement 703:713 measurement 703:713 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 4 21 theme glycans 727:733 arg1 release 691:697 in situ PNGase F-mediated release 665:697 in situ PNGase F-mediated release 665:697 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 22 theme higher 1254:1259 arg1 resolution 1261:1270 a higher resolution 1252:1270 a higher resolution 1252:1270 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 6 23 theme tissue-specific 1377:1391 arg1 N-glycans 1393:1401 tissue-specific N-glycans 1377:1401 tissue-specific N-glycans 1377:1401 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 2 24 theme analysis 296:303 arg1 type 288:291 This type 283:291 This type of analysis 283:303 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 2 24 theme analysis 296:303 arg1 imaging 384:390 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 1 25 theme mass 190:193 arg1 spectrometry 195:206 in situ mass spectrometry 182:206 retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples 168:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 1 25 theme mass 190:193 arg1 MS 209:210 MS 209:210 MS 209:210 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 0 26 theme imaging 6:12 arg1 spectrometry 19:30 MALDI imaging mass spectrometry 0:30 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney 0:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 4 27 theme present 552:558 arg1 study 560:564 the present study 548:564 the present study 548:564 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 2 28 theme matrix-assisted 332:346 arg1 MALDI 377:381 MALDI 377:381 MALDI 377:381 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 2 28 theme matrix-assisted 332:346 arg1 desorption/ionization 354:374 matrix-assisted laser desorption/ionization 332:374 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 1 29 theme Recent 103:108 arg1 developments 110:121 Recent developments 103:121 Recent developments in spatial proteomics 103:143 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 1 30 theme spectrometry 195:206 arg1 analyses 213:220 retrospective in situ mass spectrometry (MS) analyses 168:220 retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples 168:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 0 31 theme MALDI 0:4 arg1 spectrometry 19:30 MALDI imaging mass spectrometry 0:30 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney 0:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 0 32 link N-linked 35:42 arg1 glycans 44:50 N-linked glycans 35:50 N-linked glycans 35:50 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 1 33 dep in 182:183 arg1 situ 185:188 situ 185:188 situ 185:188 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 3 34 theme formalin-fixed paraffin-embedded 403:434 arg1 analyses 450:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 3 35 theme profiles 535:542 arg1 analyses 491:498 in situ analyses 483:498 in situ analyses of tissue-specific N-glycosylation profiles 483:542 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 0 36 theme mass 14:17 arg1 spectrometry 19:30 MALDI imaging mass spectrometry 0:30 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney 0:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 4 37 theme MALDI 639:643 arg1 imaging 645:651 N-glycan MALDI imaging 630:651 N-glycan MALDI imaging 630:651 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 38 dep extraction 941:950 arg1 i 938:938 i 938:938 i 938:938 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 1 39 theme spatial 126:132 arg1 proteomics 134:143 spatial proteomics 126:143 spatial proteomics 126:143 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 4 40 theme N-linked 718:725 arg1 glycans 727:733 N-linked glycans 718:733 N-linked glycans 718:733 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 6 41 theme MALDI 1339:1343 arg1 imaging 1345:1351 MALDI imaging 1339:1351 MALDI imaging 1339:1351 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 0 42 theme N-linked 35:42 arg1 glycans 44:50 N-linked glycans 35:50 N-linked glycans 35:50 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 4 43 theme N-glycan 630:637 arg1 imaging 645:651 N-glycan MALDI imaging 630:651 N-glycan MALDI imaging 630:651 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 3 44 dep in 483:484 arg1 situ 486:489 situ 486:489 situ 486:489 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 3 45 theme tissue-specific 503:517 arg1 profiles 535:542 tissue-specific N-glycosylation profiles 503:542 tissue-specific N-glycosylation profiles 503:542 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 5 46 dep profiling 1089:1097 arg1 ii 1079:1080 ii 1079:1080 ii 1079:1080 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 4 47 theme automated 590:598 arg1 method 619:624 an improved automated sample preparation method 578:624 an improved automated sample preparation method 578:624 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 48 theme presented 797:805 arg1 data 807:810 the presented data 793:810 the presented data 793:810 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 4 49 theme F-mediated 680:689 arg1 release 691:697 in situ PNGase F-mediated release 665:697 in situ PNGase F-mediated release 665:697 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 50 theme tissue 1217:1222 arg1 specificity 1224:1234 the tissue specificity 1213:1234 the tissue specificity of N-glycans at a higher resolution 1213:1270 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 3 51 theme N-glycosylation 519:533 arg1 profiles 535:542 tissue-specific N-glycosylation profiles 503:542 tissue-specific N-glycosylation profiles 503:542 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 4 52 theme improved 581:588 arg1 method 619:624 an improved automated sample preparation method 578:624 an improved automated sample preparation method 578:624 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 53 from resolution 1261:1270 arg1 specificity 1224:1234 the tissue specificity 1213:1234 the tissue specificity of N-glycans at a higher resolution 1213:1270 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 54 theme large 1147:1151 arg1 arrays 1161:1166 large droplet arrays 1147:1166 large droplet arrays in situ 1147:1174 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 55 theme on-target 985:993 arg1 MS 1001:1002 on-target MALDI MS 985:1002 on-target MALDI MS 985:1002 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 0 56 theme glycans 44:50 arg1 spectrometry 19:30 MALDI imaging mass spectrometry 0:30 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney 0:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 4 57 theme preparation 607:617 arg1 method 619:624 an improved automated sample preparation method 578:624 an improved automated sample preparation method 578:624 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 4 58 link N-linked 718:725 arg1 glycans 727:733 N-linked glycans 718:733 N-linked glycans 718:733 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 59 theme MALDI 995:999 arg1 MS 1001:1002 on-target MALDI MS 985:1002 on-target MALDI MS 985:1002 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 4 60 theme sample 600:605 arg1 method 619:624 an improved automated sample preparation method 578:624 an improved automated sample preparation method 578:624 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 61 dep strategies 925:934 arg1 analysis 968:975 composition analysis 956:975 composition analysis 956:975 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 61 dep strategies 925:934 arg1 strategies 925:934 three strategies 919:934 three strategies: (i) extraction and composition analysis 919:975 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 61 dep strategies 925:934 arg1 extraction 941:950 (i) extraction 937:950 (i) extraction 937:950 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 62 theme MALDI 1083:1087 arg1 profiling 1089:1097 (ii) MALDI profiling 1078:1097 (ii) MALDI profiling 1078:1097 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 63 theme MALDI 1187:1191 arg1 imaging 1193:1199 (iii) MALDI imaging 1181:1199 (iii) MALDI imaging 1181:1199 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 1 64 theme formalin-fixed paraffin-embedded 225:256 arg1 samples 274:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 5 65 theme trap 1069:1072 arg1 MS 1074:1075 electrospray ionization ion trap MS 1041:1075 electrospray ionization ion trap MS 1041:1075 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 5 66 theme electrospray 1041:1052 arg1 MS 1074:1075 electrospray ionization ion trap MS 1041:1075 electrospray ionization ion trap MS 1041:1075 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 2 67 theme desorption/ionization 354:374 arg1 type 288:291 This type 283:291 This type of analysis 283:303 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 2 67 theme desorption/ionization 354:374 arg1 imaging 384:390 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 matrix-assisted laser desorption/ionization (MALDI) imaging 332:390 This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. 25434632 1 68 theme clinical 258:265 arg1 samples 274:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 0 69 theme murine 88:93 arg1 kidney 95:100 formalin-fixed paraffin-embedded murine kidney 55:100 formalin-fixed paraffin-embedded murine kidney 55:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 4 70 dep in 665:666 arg1 situ 668:671 situ 668:671 situ 668:671 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 5 71 dep using 913:917 arg1 maps 1208:1211 maps 1208:1211 maps the tissue specificity of N-glycans at a higher resolution 1208:1270 The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. 25434632 4 72 theme formalin-fixed 752:765 arg1 kidney 774:779 formalin-fixed murine kidney 752:779 formalin-fixed murine kidney 752:779 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 1 73 from developments 110:121 arg1 proteomics 134:143 spatial proteomics 126:143 spatial proteomics 126:143 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 1 74 theme tissue 267:272 arg1 samples 274:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 formalin-fixed paraffin-embedded clinical tissue samples 225:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 25434632 0 75 theme formalin-fixed paraffin-embedded 55:86 arg1 kidney 95:100 formalin-fixed paraffin-embedded murine kidney 55:100 formalin-fixed paraffin-embedded murine kidney 55:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 0 76 from spectrometry 19:30 arg1 kidney 95:100 formalin-fixed paraffin-embedded murine kidney 55:100 formalin-fixed paraffin-embedded murine kidney 55:100 MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney. 25434632 3 77 theme MALDI 436:440 arg1 analyses 450:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 4 78 theme in 665:666 arg1 release 691:697 in situ PNGase F-mediated release 665:697 in situ PNGase F-mediated release 665:697 In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. 25434632 3 79 theme imaging 442:448 arg1 analyses 450:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 formalin-fixed paraffin-embedded MALDI imaging analyses 403:457 Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. 25434632 6 80 theme complete 1292:1299 arg1 method 1318:1323 a complete, straightforward method 1290:1323 a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies 1290:1437 Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies. 25434632 1 81 theme samples 274:280 arg1 analyses 213:220 retrospective in situ mass spectrometry (MS) analyses 168:220 retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples 168:280 Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. 24881670 6 0 theme source 1059:1064 arg1 fragmentation 1066:1078 source fragmentation 1059:1078 source fragmentation in addition to high specificity feature LC-MS detection 1059:1134 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 4 1 theme traditional 685:695 arg1 processing 708:717 traditional LC-MS data processing 685:717 traditional LC-MS data processing 685:717 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 9 2 theme serum 1553:1557 arg1 profile 1568:1574 a broad human serum N-glycan profile 1539:1574 a broad human serum N-glycan profile 1539:1574 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 3 3 from sets 517:520 arg1 critical 490:497 critical 490:497 critical 490:497 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 4 4 theme data 703:706 arg1 processing 708:717 traditional LC-MS data processing 685:717 traditional LC-MS data processing 685:717 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 10 5 theme glycan 1732:1737 arg1 isomers 1739:1745 640 glycan isomers 1728:1745 640 glycan isomers 1728:1745 A total of 156 glycan compositions and 640 glycan isomers were detected from a single sample. 24881670 11 6 theme assignments 1822:1832 arg1 assignments 1822:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 11 6 theme assignments 1822:1832 arg1 % 1790:1790 Over 99% 1783:1790 Over 99% of the GlyQ-IQ glycan-feature assignments 1783:1832 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 5 7 theme elemental 778:786 arg1 composition 788:798 the individual target's elemental composition 754:798 the individual target's elemental composition 754:798 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 6 8 theme family 1031:1036 arg1 relationships 1038:1050 glycan family relationships 1024:1050 glycan family relationships 1024:1050 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 11 9 theme GlyQ-IQ 1799:1805 arg1 assignments 1822:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 2 10 theme data 327:330 arg1 sets 332:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 11 used used 1522:1525 arg2 GlyQ-IQ 1510:1516 GlyQ-IQ 1510:1516 GlyQ-IQ 1510:1516 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 12 theme resolution 1588:1597 arg1 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 12 theme resolution 1588:1597 arg1 spectrometry 1650:1661 high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry 1583:1661 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 6 13 theme LC-MS 1120:1124 arg1 detection 1126:1134 high specificity feature LC-MS detection 1095:1134 high specificity feature LC-MS detection 1095:1134 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 8 14 theme confounding 1406:1416 arg1 peaks 1434:1438 confounding chromatographic peaks 1406:1438 confounding chromatographic peaks 1406:1438 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 2 15 theme extracted 353:361 arg1 chromatograms 367:379 convoluted extracted ion chromatograms 342:379 convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools 342:443 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 16 theme chromatography-tandem 1623:1643 arg1 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 16 theme chromatography-tandem 1623:1643 arg1 spectrometry 1650:1661 high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry 1583:1661 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 6 17 theme specificity 1100:1110 arg1 detection 1126:1134 high specificity feature LC-MS detection 1095:1134 high specificity feature LC-MS detection 1095:1134 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 5 18 theme isotopic 943:950 arg1 species 952:958 the isotopic species 939:958 the isotopic species having the highest intensity 939:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 9 19 theme spectrometry 1650:1661 arg1 set 1684:1686 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 8 20 theme high 1445:1448 arg1 isomers 1465:1471 high mannose glycan isomers 1445:1471 high mannose glycan isomers from human blood serum 1445:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 10 21 theme single 1768:1773 arg1 sample 1775:1780 a single sample 1766:1780 a single sample 1766:1780 A total of 156 glycan compositions and 640 glycan isomers were detected from a single sample. 24881670 1 22 from N-glycans 244:252 arg1 data 305:308 liquid chromatography-mass spectrometry (LC-MS) data 257:308 liquid chromatography-mass spectrometry (LC-MS) data 257:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 8 23 theme glycan 1458:1463 arg1 isomers 1465:1471 high mannose glycan isomers 1445:1471 high mannose glycan isomers from human blood serum 1445:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 7 24 theme GlyQ-IQ 1184:1190 arg1 software 1192:1199 The GlyQ-IQ software 1180:1199 The GlyQ-IQ software 1180:1199 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 2 25 theme Glycomics 311:319 arg1 sets 332:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 26 theme data 1679:1682 arg1 set 1684:1686 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 6 27 theme analysis 1170:1177 arg1 specificity 1151:1161 the specificity 1147:1161 the specificity of the analysis 1147:1177 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 5 28 theme improved 811:818 arg1 sensitivity 820:830 improved sensitivity 811:830 improved sensitivity 811:830 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 8 29 theme blood 1484:1488 arg1 serum 1490:1494 human blood serum 1478:1494 human blood serum 1478:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 0 30 theme 4D 103:104 arg1 visualization 106:118 GlycoGrid 4D visualization 93:118 GlycoGrid 4D visualization 93:118 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 1 31 theme glycomics 204:212 arg1 tool 223:226 a biologically guided glycomics analysis tool 182:226 a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data 182:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 31 theme glycomics 204:212 arg1 quantification 154:167 Glycomics quintavariate-informed quantification 121:167 Glycomics quintavariate-informed quantification (GlyQ-IQ) 121:177 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 5 32 theme isotope 855:861 arg1 information 871:881 the exact isotope profile information 845:881 the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity 845:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 3 33 theme data 512:515 arg1 sets 517:520 glycomics data sets 502:520 glycomics data sets 502:520 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 7 34 from sets 1324:1327 arg1 compositions 1283:1294 the N-glycan compositions 1270:1294 the N-glycan compositions from human serum LC-MS data sets 1270:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 1 35 theme Glycomics 121:129 arg1 tool 223:226 a biologically guided glycomics analysis tool 182:226 a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data 182:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 35 theme Glycomics 121:129 arg1 GlyQ-IQ 170:176 GlyQ-IQ 170:176 GlyQ-IQ 170:176 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 35 theme Glycomics 121:129 arg1 quantification 154:167 Glycomics quintavariate-informed quantification 121:167 Glycomics quintavariate-informed quantification (GlyQ-IQ) 121:177 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 5 36 contain having 960:965 arg2 intensity 979:987 the highest intensity 967:987 the highest intensity 967:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 5 36 contain having 960:965 arg1 species 952:958 the isotopic species 939:958 the isotopic species having the highest intensity 939:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 7 37 theme LC-MS 1313:1317 arg1 sets 1324:1327 human serum LC-MS data sets 1301:1327 human serum LC-MS data sets 1301:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 2 38 contain have 337:340 arg2 chromatograms 367:379 convoluted extracted ion chromatograms 342:379 convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools 342:443 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 2 38 contain have 337:340 arg1 sets 332:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 4 39 theme key 667:669 arg1 advantages 671:680 several key advantages 659:680 several key advantages 659:680 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 7 40 theme human 1301:1305 arg1 sets 1324:1327 human serum LC-MS data sets 1301:1327 human serum LC-MS data sets 1301:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 5 41 theme LC 920:921 arg1 peak 923:926 LC peak 920:926 LC peak 920:926 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 6 42 theme Glycan 990:995 arg1 annotation 1004:1013 Glycan target annotation 990:1013 Glycan target annotation 990:1013 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 3 43 theme constituent 468:478 arg1 pieces 480:485 constituent pieces 468:485 constituent pieces 468:485 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 4 44 theme biological 614:623 arg1 approach 643:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 3 45 theme intact 576:581 arg1 isomers 601:607 different intact glycan structural isomers 566:607 different intact glycan structural isomers 566:607 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 0 46 theme high-performance 62:77 arg1 computing 79:87 high-performance computing 62:87 high-performance computing 62:87 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 4 47 theme analysis 634:641 arg1 approach 643:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 11 48 theme mass 1895:1898 arg1 spectra 1900:1906 high-resolution mass spectra 1879:1906 high-resolution mass spectra 1879:1906 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 1 49 theme spectrometry 284:295 arg1 data 305:308 liquid chromatography-mass spectrometry (LC-MS) data 257:308 liquid chromatography-mass spectrometry (LC-MS) data 257:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 10 50 theme compositions 1711:1722 arg1 total 1691:1695 A total 1689:1695 A total of 156 glycan compositions and 640 glycan isomers 1689:1745 A total of 156 glycan compositions and 640 glycan isomers were detected from a single sample. 24881670 8 51 from serum 1490:1494 arg1 isomers 1465:1471 high mannose glycan isomers 1445:1471 high mannose glycan isomers from human blood serum 1445:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 0 52 dep GlyQ-IQ 0:6 arg1 quantification 42:55 glycomics quintavariate-informed quantification 9:55 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 0:119 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 5 53 theme individual 758:767 arg1 target 769:774 the individual target's 754:776 the individual target's elemental composition 754:798 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 0 54 with quantification 42:55 arg1 visualization 106:118 GlycoGrid 4D visualization 93:118 GlycoGrid 4D visualization 93:118 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 0 54 with quantification 42:55 arg1 computing 79:87 high-performance computing 62:87 high-performance computing 62:87 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 3 55 theme structural 590:599 arg1 isomers 601:607 different intact glycan structural isomers 566:607 different intact glycan structural isomers 566:607 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 9 56 theme human 1547:1551 arg1 profile 1568:1574 a broad human serum N-glycan profile 1539:1574 a broad human serum N-glycan profile 1539:1574 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 10 57 theme isomers 1739:1745 arg1 total 1691:1695 A total 1689:1695 A total of 156 glycan compositions and 640 glycan isomers 1689:1745 A total of 156 glycan compositions and 640 glycan isomers were detected from a single sample. 24881670 2 58 theme convoluted 342:351 arg1 chromatograms 367:379 convoluted extracted ion chromatograms 342:379 convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools 342:443 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 59 theme N-glycan 1559:1566 arg1 profile 1568:1574 a broad human serum N-glycan profile 1539:1574 a broad human serum N-glycan profile 1539:1574 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 4 60 theme LC-MS 697:701 arg1 processing 708:717 traditional LC-MS data processing 685:717 traditional LC-MS data processing 685:717 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 5 61 theme glycan 729:734 arg1 information 736:746 A priori glycan information 720:746 A priori glycan information about the individual target's elemental composition 720:798 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 1 62 theme analysis 214:221 arg1 tool 223:226 a biologically guided glycomics analysis tool 182:226 a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data 182:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 62 theme analysis 214:221 arg1 quantification 154:167 Glycomics quintavariate-informed quantification 121:167 Glycomics quintavariate-informed quantification (GlyQ-IQ) 121:177 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 6 63 theme glycan 1024:1029 arg1 relationships 1038:1050 glycan family relationships 1024:1050 glycan family relationships 1024:1050 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 11 64 theme glycan-feature 1807:1820 arg1 assignments 1822:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 the GlyQ-IQ glycan-feature assignments 1795:1832 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 9 65 theme high 1583:1586 arg1 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 65 theme high 1583:1586 arg1 spectrometry 1650:1661 high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry 1583:1661 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 2 66 theme ion 363:365 arg1 chromatograms 367:379 convoluted extracted ion chromatograms 342:379 convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools 342:443 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 67 theme nanoelectrospray-liquid 1599:1621 arg1 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 67 theme nanoelectrospray-liquid 1599:1621 arg1 spectrometry 1650:1661 high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry 1583:1661 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 8 68 theme case 1332:1335 arg1 study 1337:1341 A case study 1330:1341 A case study 1330:1341 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 6 69 theme feature 1112:1118 arg1 detection 1126:1134 high specificity feature LC-MS detection 1095:1134 high specificity feature LC-MS detection 1095:1134 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 5 70 theme highest 971:977 arg1 intensity 979:987 the highest intensity 967:987 the highest intensity 967:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 6 71 theme high 1095:1098 arg1 detection 1126:1134 high specificity feature LC-MS detection 1095:1134 high specificity feature LC-MS detection 1095:1134 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 8 72 theme chromatographic 1418:1432 arg1 peaks 1434:1438 confounding chromatographic peaks 1406:1438 confounding chromatographic peaks 1406:1438 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 7 73 theme serum 1307:1311 arg1 sets 1324:1327 human serum LC-MS data sets 1301:1327 human serum LC-MS data sets 1301:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 2 74 theme LC-MS 321:325 arg1 sets 332:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets 311:335 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 6 75 dep detection 1126:1134 arg1 addition 1083:1090 addition 1083:1090 addition 1083:1090 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 8 76 theme mannose 1450:1456 arg1 isomers 1465:1471 high mannose glycan isomers 1445:1471 high mannose glycan isomers from human blood serum 1445:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 0 77 theme GlycoGrid 93:101 arg1 visualization 106:118 GlycoGrid 4D visualization 93:118 GlycoGrid 4D visualization 93:118 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 5 78 theme chromatogram 892:903 arg1 generation 905:914 chromatogram generation 892:914 chromatogram generation 892:914 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 1 79 theme guided 197:202 arg1 tool 223:226 a biologically guided glycomics analysis tool 182:226 a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data 182:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 79 theme guided 197:202 arg1 quantification 154:167 Glycomics quintavariate-informed quantification 121:167 Glycomics quintavariate-informed quantification (GlyQ-IQ) 121:177 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 8 80 theme human 1478:1482 arg1 serum 1490:1494 human blood serum 1478:1494 human blood serum 1478:1494 A case study is presented to demonstrate how GlyQ-IQ identifies and removes confounding chromatographic peaks from high mannose glycan isomers from human blood serum. 24881670 5 81 theme exact 849:853 arg1 information 871:881 the exact isotope profile information 845:881 the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity 845:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 3 82 from critical 490:497 arg1 sets 517:520 glycomics data sets 502:520 glycomics data sets 502:520 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 5 83 theme profile 863:869 arg1 information 871:881 the exact isotope profile information 845:881 the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity 845:987 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 5 84 theme priori 722:727 arg1 information 736:746 A priori glycan information 720:746 A priori glycan information about the individual target's elemental composition 720:798 A priori glycan information about the individual target's elemental composition allows for improved sensitivity by utilizing the exact isotope profile information to focus chromatogram generation and LC peak fitting on the isotopic species having the highest intensity. 24881670 1 85 theme liquid 257:262 arg1 LC-MS 298:302 LC-MS 298:302 LC-MS 298:302 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 85 theme liquid 257:262 arg1 spectrometry 284:295 liquid chromatography-mass spectrometry 257:295 liquid chromatography-mass spectrometry (LC-MS) data 257:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 7 86 theme data 1319:1322 arg1 sets 1324:1327 human serum LC-MS data sets 1301:1327 human serum LC-MS data sets 1301:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 4 87 theme several 659:665 arg1 advantages 671:680 several key advantages 659:680 several key advantages 659:680 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 3 88 theme glycomics 502:510 arg1 sets 517:520 glycomics data sets 502:520 glycomics data sets 502:520 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 0 89 theme quintavariate-informed 19:40 arg1 quantification 42:55 glycomics quintavariate-informed quantification 9:55 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 0:119 GlyQ-IQ: glycomics quintavariate-informed quantification with high-performance computing and GlycoGrid 4D visualization. 24881670 1 90 theme quintavariate-informed 131:152 arg1 tool 223:226 a biologically guided glycomics analysis tool 182:226 a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data 182:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 90 theme quintavariate-informed 131:152 arg1 GlyQ-IQ 170:176 GlyQ-IQ 170:176 GlyQ-IQ 170:176 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 90 theme quintavariate-informed 131:152 arg1 quantification 154:167 Glycomics quintavariate-informed quantification 121:167 Glycomics quintavariate-informed quantification (GlyQ-IQ) 121:177 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 3 91 theme LC 446:447 arg1 deconvolution 449:461 LC deconvolution 446:461 LC deconvolution into constituent pieces 446:485 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 6 92 theme target 997:1002 arg1 annotation 1004:1013 Glycan target annotation 990:1013 Glycan target annotation 990:1013 Glycan target annotation utilizes glycan family relationships and in source fragmentation in addition to high specificity feature LC-MS detection to improve the specificity of the analysis. 24881670 4 93 theme targeted 625:632 arg1 approach 643:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach 610:650 The biological targeted analysis approach offers several key advantages to traditional LC-MS data processing. 24881670 11 94 theme manual 1841:1846 arg1 validation 1848:1857 manual validation 1841:1857 manual validation 1841:1857 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 3 95 theme chromatographic 530:544 arg1 peaks 546:550 chromatographic peaks 530:550 chromatographic peaks 530:550 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 7 96 theme N-glycan 1274:1281 arg1 compositions 1283:1294 the N-glycan compositions 1270:1294 the N-glycan compositions from human serum LC-MS data sets 1270:1327 The GlyQ-IQ software was developed in this work and evaluated in the context of profiling the N-glycan compositions from human serum LC-MS data sets. 24881670 9 97 theme mass 1645:1648 arg1 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 nESI-LC-MS/MS 1664:1676 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 9 97 theme mass 1645:1648 arg1 spectrometry 1650:1661 high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry 1583:1661 a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set 1581:1686 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 24881670 11 98 theme high-resolution 1879:1893 arg1 spectra 1900:1906 high-resolution mass spectra 1879:1906 high-resolution mass spectra 1879:1906 Over 99% of the GlyQ-IQ glycan-feature assignments passed manual validation and are backed with high-resolution mass spectra. 24881670 3 99 theme glycan 583:588 arg1 isomers 601:607 different intact glycan structural isomers 566:607 different intact glycan structural isomers 566:607 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 3 100 theme different 566:574 arg1 isomers 601:607 different intact glycan structural isomers 566:607 different intact glycan structural isomers 566:607 LC deconvolution into constituent pieces is critical in glycomics data sets because chromatographic peaks correspond to different intact glycan structural isomers. 24881670 1 101 theme chromatography-mass 264:282 arg1 LC-MS 298:302 LC-MS 298:302 LC-MS 298:302 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 1 101 theme chromatography-mass 264:282 arg1 spectrometry 284:295 liquid chromatography-mass spectrometry 257:295 liquid chromatography-mass spectrometry (LC-MS) data 257:308 Glycomics quintavariate-informed quantification (GlyQ-IQ) is a biologically guided glycomics analysis tool for identifying N-glycans in liquid chromatography-mass spectrometry (LC-MS) data. 24881670 10 102 theme glycan 1704:1709 arg1 compositions 1711:1722 156 glycan compositions 1700:1722 156 glycan compositions 1700:1722 A total of 156 glycan compositions and 640 glycan isomers were detected from a single sample. 24881670 2 103 theme software 430:437 arg1 tools 439:443 software tools 430:443 software tools 430:443 Glycomics LC-MS data sets have convoluted extracted ion chromatograms that are challenging to deconvolve with existing software tools. 24881670 9 104 theme broad 1541:1545 arg1 profile 1568:1574 a broad human serum N-glycan profile 1539:1574 a broad human serum N-glycan profile 1539:1574 In addition, GlyQ-IQ was used to generate a broad human serum N-glycan profile from a high resolution nanoelectrospray-liquid chromatography-tandem mass spectrometry (nESI-LC-MS/MS) data set. 25071157 0 0 theme immunoglobulin 75:88 arg1 IgA 93:95 IgA 93:95 IgA 93:95 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 0 theme immunoglobulin 75:88 arg1 A 90:90 immunoglobulin A 75:90 immunoglobulin A (IgA) nephropathy 75:108 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 1 1 theme glomerular 324:333 arg1 mesangium 335:343 the glomerular mesangium 320:343 the glomerular mesangium 320:343 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 3 2 with patients 794:801 arg1 IgAN 808:811 IgAN 808:811 IgAN 808:811 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 1 3 theme A 150:150 arg1 form 190:193 the most common form 174:193 the most common form of glomerulonephritis worldwide 174:225 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 3 theme A 150:150 arg1 IgAN 165:168 IgAN 165:168 IgAN 165:168 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 3 theme A 150:150 arg1 nephropathy 152:162 Immunoglobulin A nephropathy 135:162 Immunoglobulin A nephropathy (IgAN) 135:169 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 6 4 theme O-glycans 1312:1320 arg1 undergalactosylation 1288:1307 undergalactosylation 1288:1307 undergalactosylation of O-glycans in IgA1 1288:1328 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 3 5 gly glycoforms 760:769 arg1 IgA1 774:777 IgA1 774:777 IgA1 774:777 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 4 6 theme plasma 853:858 arg1 IgA 860:862 total plasma IgA 847:862 total plasma IgA in IgAN patients 847:879 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 2 7 theme Prior 346:350 arg1 studies 352:358 Prior studies 346:358 Prior studies 346:358 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 3 8 theme healthy 817:823 arg1 individuals 825:835 healthy individuals 817:835 healthy individuals 817:835 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 4 9 theme healthy 927:933 arg1 donors 935:940 healthy donors 927:940 healthy donors 927:940 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 0 10 from Identification 0:13 arg1 individuals 122:132 healthy individuals 114:132 healthy individuals 114:132 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 10 from Identification 0:13 arg1 patients 61:68 patients 61:68 patients with immunoglobulin A (IgA) nephropathy 61:108 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 10 from Identification 0:13 arg1 plasma 49:54 plasma 49:54 plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals 49:132 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 4 11 theme IgAN 867:870 arg1 patients 872:879 IgAN patients 867:879 IgAN patients 867:879 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 6 12 theme cells 1454:1458 arg1 subpopulation 1425:1437 a subpopulation 1423:1437 a subpopulation of B or plasma cells 1423:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 6 13 from undergalactosylation 1288:1307 arg1 IgA1 1325:1328 IgA1 1325:1328 IgA1 1325:1328 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 4 14 theme core 1029:1032 arg1 O-glycans 1042:1050 core 1 based O-glycans 1029:1050 core 1 based O-glycans 1029:1050 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 0 15 theme A 90:90 arg1 nephropathy 98:108 immunoglobulin A (IgA) nephropathy 75:108 immunoglobulin A (IgA) nephropathy 75:108 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 3 16 with individuals 825:835 arg1 IgAN 808:811 IgAN 808:811 IgAN 808:811 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 5 17 theme core 1206:1209 arg1 structure 1213:1221 the core 1 structure 1202:1221 the core 1 structure 1202:1221 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 0 18 with patients 61:68 arg1 nephropathy 98:108 immunoglobulin A (IgA) nephropathy 75:108 immunoglobulin A (IgA) nephropathy 75:108 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 4 19 theme elevated 885:892 arg1 ∼ 894:894 elevated ∼ 885:894 elevated ∼ 1.6-fold compared with that in healthy donors 885:940 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 3 20 from individuals 825:835 arg1 plasma 782:787 plasma 782:787 plasma from patients with IgAN and healthy individuals 782:835 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 6 21 theme IgA 1493:1495 arg1 elevation 1480:1488 overall elevation 1472:1488 overall elevation of IgA 1472:1495 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 6 21 theme IgA 1493:1495 arg1 inefficiency 1381:1392 in vivo inefficiency 1373:1392 in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells 1373:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 4 22 theme Tn/STn 1090:1095 arg1 structures 1097:1106 Tn/STn structures 1090:1106 Tn/STn structures 1090:1106 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 2 23 theme IgAN 391:394 arg1 patients 396:403 IgAN patients 391:403 IgAN patients 391:403 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 4 24 dep glycoforms 1008:1017 arg1 one 1020:1022 one 1020:1022 one 1020:1022 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 4 24 dep glycoforms 1008:1017 arg1 other 1061:1065 other 1061:1065 other 1061:1065 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 3 25 theme different 750:758 arg1 glycoforms 760:769 different glycoforms 750:769 different glycoforms of IgA1 750:777 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 4 26 theme distinct 999:1006 arg1 glycoforms 1008:1017 two distinct glycoforms 995:1017 two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures 995:1106 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 3 27 theme mass 703:706 arg1 spectrometry 708:719 mass spectrometry 703:719 mass spectrometry 703:719 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 4 28 theme total 847:851 arg1 IgA 860:862 total plasma IgA 847:862 total plasma IgA in IgAN patients 847:879 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 2 29 gly sialylated 487:496 arg1 version 498:504 its sialylated version 483:504 its sialylated version 483:504 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 6 30 theme overall 1472:1478 arg1 elevation 1480:1488 overall elevation 1472:1488 overall elevation of IgA 1472:1495 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 4 31 from IgA 860:862 arg1 patients 872:879 IgAN patients 867:879 IgAN patients 867:879 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 1 32 from deposition 270:279 arg1 mesangium 335:343 the glomerular mesangium 320:343 the glomerular mesangium 320:343 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 2 33 theme serum 375:379 arg1 IgA1 381:384 serum IgA1 375:384 serum IgA1 from IgAN patients 375:403 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 0 34 theme glycoforms 27:36 arg1 Identification 0:13 Identification 0:13 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 0:133 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 3 35 used used 616:619 arg2 we 608:609 we 608:609 we 608:609 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 5 36 from controls 1172:1179 arg1 IgA1 1144:1147 IgA1 1144:1147 IgA1 from IgAN patients and controls 1144:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 2 37 dep aberrant 414:421 arg1 undergalactosylated 424:442 undergalactosylated 424:442 undergalactosylated 424:442 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 5 38 attach present 1133:1139 arg1 IgA1 1144:1147 IgA1 1144:1147 IgA1 from IgAN patients and controls 1144:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 5 38 attach present 1133:1139 arg2 antigen 1125:1131 Tn antigen 1122:1131 Tn antigen present on IgA1 from IgAN patients and controls 1122:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 0 39 theme distinct 18:25 arg1 glycoforms 27:36 distinct glycoforms 18:36 distinct glycoforms of IgA1 18:44 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 4 40 contain containing 1079:1088 arg2 structures 1097:1106 Tn/STn structures 1090:1106 Tn/STn structures 1090:1106 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 4 40 contain containing 1079:1088 arg1 other 1061:1065 other 1061:1065 other 1061:1065 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 3 41 theme lectin 628:633 arg1 technologies 646:657 serial lectin separation technologies 621:657 serial lectin separation technologies 621:657 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 2 42 theme aberrant 414:421 arg1 O-glycans 444:452 aberrant, undergalactosylated O-glycans 414:452 aberrant, undergalactosylated O-glycans 414:452 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 0 43 theme IgA1 41:44 arg1 glycoforms 27:36 distinct glycoforms 18:36 distinct glycoforms of IgA1 18:44 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 1 44 theme IgA1 284:287 arg1 deposition 270:279 the deposition 266:279 the deposition of IgA1 and consequent inflammation in the glomerular mesangium 266:343 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 5 45 from patients 1159:1166 arg1 IgA1 1144:1147 IgA1 1144:1147 IgA1 from IgAN patients and controls 1144:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 3 46 theme separation 635:644 arg1 technologies 646:657 serial lectin separation technologies 621:657 serial lectin separation technologies 621:657 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 6 47 theme B 1442:1442 arg1 cells 1454:1458 B or plasma cells 1442:1458 cells 1454:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 1 48 theme common 183:188 arg1 nephropathy 152:162 Immunoglobulin A nephropathy 135:162 Immunoglobulin A nephropathy (IgAN) 135:169 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 48 theme common 183:188 arg1 form 190:193 the most common form 174:193 the most common form of glomerulonephritis worldwide 174:225 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 2 49 theme Tn 468:469 arg1 antigen 471:477 Tn antigen 468:477 Tn antigen 468:477 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 0 50 theme healthy 114:120 arg1 individuals 122:132 healthy individuals 114:132 healthy individuals 114:132 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 51 from patients 61:68 arg1 Identification 0:13 Identification 0:13 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 0:133 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 51 from patients 61:68 arg1 plasma 49:54 plasma 49:54 plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals 49:132 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 6 52 theme T-synthase 1397:1406 arg1 elevation 1480:1488 overall elevation 1472:1488 overall elevation of IgA 1472:1495 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 6 52 theme T-synthase 1397:1406 arg1 inefficiency 1381:1392 in vivo inefficiency 1373:1392 in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells 1373:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 5 53 theme IgAN 1154:1157 arg1 patients 1159:1166 IgAN patients 1154:1166 IgAN patients 1154:1166 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 6 54 theme in 1373:1374 arg1 inefficiency 1381:1392 in vivo inefficiency 1373:1392 in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells 1373:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 1 55 theme consequent 293:302 arg1 inflammation 304:315 consequent inflammation 293:315 consequent inflammation 293:315 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 3 56 theme serial 621:626 arg1 technologies 646:657 serial lectin separation technologies 621:657 serial lectin separation technologies 621:657 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 3 57 theme enzymatic 674:682 arg1 modifications 684:696 enzymatic modifications 674:696 enzymatic modifications 674:696 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 1 58 theme inflammation 304:315 arg1 deposition 270:279 the deposition 266:279 the deposition of IgA1 and consequent inflammation in the glomerular mesangium 266:343 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 2 59 theme aberrant 553:560 arg1 O-glycosylation 562:576 aberrant O-glycosylation 553:576 aberrant O-glycosylation 553:576 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 3 60 theme Western 660:666 arg1 blot 668:671 Western blot 660:671 Western blot 660:671 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 0 61 gly glycoforms 27:36 arg1 IgA1 41:44 IgA1 41:44 IgA1 41:44 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 6 62 dep in 1373:1374 arg1 vivo 1376:1379 vivo 1376:1379 vivo 1376:1379 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 0 63 with individuals 122:132 arg1 nephropathy 98:108 immunoglobulin A (IgA) nephropathy 75:108 immunoglobulin A (IgA) nephropathy 75:108 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 5 64 theme present 1133:1139 arg1 antigen 1125:1131 Tn antigen 1122:1131 Tn antigen present on IgA1 from IgAN patients and controls 1122:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 2 65 contain contains 405:412 arg1 IgA1 381:384 serum IgA1 375:384 serum IgA1 from IgAN patients 375:403 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 2 65 contain contains 405:412 arg2 O-glycans 444:452 aberrant, undergalactosylated O-glycans 414:452 aberrant, undergalactosylated O-glycans 414:452 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 6 66 theme plasma 1447:1452 arg1 cells 1454:1458 B or plasma cells 1442:1458 cells 1454:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 6 67 theme IgAN 1516:1519 arg1 pathogenesis 1521:1532 IgAN pathogenesis 1516:1532 IgAN pathogenesis 1516:1532 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 4 68 theme based 1036:1040 arg1 O-glycans 1042:1050 core 1 based O-glycans 1029:1050 core 1 based O-glycans 1029:1050 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 0 69 from individuals 122:132 arg1 Identification 0:13 Identification 0:13 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 0:133 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 0 69 from individuals 122:132 arg1 plasma 49:54 plasma 49:54 plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals 49:132 Identification of distinct glycoforms of IgA1 in plasma from patients with immunoglobulin A (IgA) nephropathy and healthy individuals. 25071157 3 70 from patients 794:801 arg1 plasma 782:787 plasma 782:787 plasma from patients with IgAN and healthy individuals 782:835 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 25071157 6 71 from inefficiency 1381:1392 arg1 subpopulation 1425:1437 a subpopulation 1423:1437 a subpopulation of B or plasma cells 1423:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 6 72 from elevation 1480:1488 arg1 subpopulation 1425:1437 a subpopulation 1423:1437 a subpopulation of B or plasma cells 1423:1458 Our results demonstrate that undergalactosylation of O-glycans in IgA1 is not restricted to IgAN and suggest that in vivo inefficiency of T-synthase toward IgA1 in a subpopulation of B or plasma cells, as well as overall elevation of IgA, may contribute to IgAN pathogenesis. 25071157 1 73 theme glomerulonephritis 198:215 arg1 worldwide 217:225 glomerulonephritis worldwide 198:225 glomerulonephritis worldwide 198:225 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 2 74 from patients 396:403 arg1 IgA1 381:384 serum IgA1 375:384 serum IgA1 from IgAN patients 375:403 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 5 75 theme recombinant 1235:1245 arg1 T-synthase 1247:1256 recombinant T-synthase 1235:1256 recombinant T-synthase 1235:1256 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 1 76 theme worldwide 217:225 arg1 nephropathy 152:162 Immunoglobulin A nephropathy 135:162 Immunoglobulin A nephropathy (IgAN) 135:169 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 76 theme worldwide 217:225 arg1 form 190:193 the most common form 174:193 the most common form of glomerulonephritis worldwide 174:225 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 2 77 theme sialylated 487:496 arg1 version 498:504 its sialylated version 483:504 its sialylated version 483:504 Prior studies suggested that serum IgA1 from IgAN patients contains aberrant, undergalactosylated O-glycans, for example, Tn antigen and its sialylated version, SialylTn (STn), but the mechanisms underlying aberrant O-glycosylation are not well understood. 25071157 5 78 theme Tn 1122:1123 arg1 antigen 1125:1131 Tn antigen 1122:1131 Tn antigen present on IgA1 from IgAN patients and controls 1122:1179 Importantly, Tn antigen present on IgA1 from IgAN patients and controls was convertible into the core 1 structure in vitro by recombinant T-synthase. 25071157 4 79 from IgA1 943:946 arg1 samples 955:961 all samples 951:961 all samples 951:961 Although total plasma IgA in IgAN patients was elevated ∼ 1.6-fold compared with that in healthy donors, IgA1 in all samples was unexpectedly separable into two distinct glycoforms: one with core 1 based O-glycans, and the other exclusively containing Tn/STn structures. 25071157 1 80 theme Immunoglobulin 135:148 arg1 form 190:193 the most common form 174:193 the most common form of glomerulonephritis worldwide 174:225 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 80 theme Immunoglobulin 135:148 arg1 IgAN 165:168 IgAN 165:168 IgAN 165:168 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 1 80 theme Immunoglobulin 135:148 arg1 nephropathy 152:162 Immunoglobulin A nephropathy 135:162 Immunoglobulin A nephropathy (IgAN) 135:169 Immunoglobulin A nephropathy (IgAN) is the most common form of glomerulonephritis worldwide and is histologically characterized by the deposition of IgA1 and consequent inflammation in the glomerular mesangium. 25071157 3 81 theme IgA1 774:777 arg1 glycoforms 760:769 different glycoforms 750:769 different glycoforms of IgA1 750:777 Here we have used serial lectin separation technologies, Western blot, enzymatic modifications, and mass spectrometry to explore whether there are different glycoforms of IgA1 in plasma from patients with IgAN and healthy individuals. 26918528 3 0 theme oligosaccharyl 575:588 arg1 transferases 590:601 the polytopic membrane-bound oligosaccharyl transferases 546:601 the polytopic membrane-bound oligosaccharyl transferases (OTases) 546:610 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 0 theme oligosaccharyl 575:588 arg1 OTases 604:609 OTases 604:609 OTases 604:609 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 5 1 theme LRET-based 860:869 arg1 measurements 880:891 LRET-based distance measurements 860:891 LRET-based distance measurements 860:891 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 7 2 theme experimental 1493:1504 arg1 information 1506:1516 new experimental information 1489:1516 new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase 1489:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 3 3 theme transferases 590:601 arg1 interactions 530:541 the interactions 526:541 the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates 526:634 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 6 4 from approach 1370:1377 arg1 system 1392:1397 a defined system 1382:1397 a defined system 1382:1397 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 6 5 theme PglB 1215:1218 arg1 measurements 1177:1188 LRET-based measurements 1166:1188 LRET-based measurements of detergent-solubilized PglB from C. lari 1166:1231 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 6 6 with comparison 1248:1257 arg1 distances 1268:1276 the distances 1264:1276 the distances based on the previously reported the C. lari PglB crystal structure 1264:1344 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 3 7 theme partner 617:623 arg1 substrates 625:634 partner substrates 617:634 partner substrates 617:634 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 7 8 theme related 1546:1552 arg1 OTase 1600:1604 the related, but structurally uncharacterized, eukaryotic OTase 1542:1604 the related, but structurally uncharacterized, eukaryotic OTase 1542:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 5 9 theme labeled 1055:1061 arg1 peptide 1063:1069 fluorescently labeled peptide and glycan substrates 1041:1091 peptide 1063:1069 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 10 theme glycan 1075:1080 arg1 substrates 1082:1091 fluorescently labeled peptide and glycan substrates 1041:1091 substrates 1082:1091 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 4 11 theme diphosphate-linked 790:807 arg1 substrates 809:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 7 12 theme Distance 1400:1407 arg1 measurements 1409:1420 Distance measurements 1400:1420 Distance measurements between peptide and glycan substrates and the C. jejuni PglB 1400:1481 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 6 13 theme defined 1384:1390 arg1 system 1392:1397 a defined system 1382:1397 a defined system 1382:1397 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 6 14 theme LRET-based 1166:1175 arg1 measurements 1177:1188 LRET-based measurements 1166:1188 LRET-based measurements of detergent-solubilized PglB from C. lari 1166:1231 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 1 15 theme major 181:185 arg1 hurdles 197:203 major technical hurdles 181:203 major technical hurdles 181:203 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 0 16 theme Luminescence 61:72 arg1 Transfer 91:98 Luminescence Resonance Energy Transfer 61:98 Luminescence Resonance Energy Transfer 61:98 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 7 17 theme eukaryotic 1589:1598 arg1 OTase 1600:1604 the related, but structurally uncharacterized, eukaryotic OTase 1542:1604 the related, but structurally uncharacterized, eukaryotic OTase 1542:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 6 18 theme direct 1241:1246 arg1 comparison 1248:1257 direct comparison 1241:1257 direct comparison with the distances based on the previously reported the C. lari PglB crystal structure 1241:1344 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 1 19 theme Integral 101:108 arg1 proteins 119:126 Integral membrane proteins 101:126 Integral membrane proteins 101:126 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 6 20 theme detergent-solubilized 1193:1213 arg1 PglB 1215:1218 detergent-solubilized PglB 1193:1218 detergent-solubilized PglB 1193:1218 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 1 21 theme technical 187:195 arg1 hurdles 197:203 major technical hurdles 181:203 major technical hurdles 181:203 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 0 22 theme Polytopic 8:16 arg1 Interactions 45:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 1 23 theme membrane 110:117 arg1 proteins 119:126 Integral membrane proteins 101:126 Integral membrane proteins 101:126 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 4 24 from transfer 732:739 arg1 substrates 809:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 3 25 theme transfer 492:499 arg1 application 447:457 the application 443:457 the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates 443:634 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 5 26 theme encoded 928:934 arg1 LBT 971:973 LBT 971:973 LBT 971:973 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 26 theme encoded 928:934 arg1 tag 966:968 an encoded N-terminal lanthanide-binding tag 925:968 an encoded N-terminal lanthanide-binding tag (LBT) 925:974 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 4 27 theme Monomeric 637:645 arg1 PglBs 667:671 the PglBs 663:671 the PglBs from Campylobacter jejuni and Campylobacter lari 663:720 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 4 27 theme Monomeric 637:645 arg1 OTases 647:652 Monomeric OTases 637:652 Monomeric OTases 637:652 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 2 28 theme determined 352:361 arg1 structures 393:402 experimentally determined and computationally predicted structures 337:402 experimentally determined and computationally predicted structures 337:402 Biophysical approaches that provide structural information to complement and leverage experimentally determined and computationally predicted structures are urgently needed. 26918528 7 29 theme peptide 1430:1436 arg1 substrates 1449:1458 peptide and glycan substrates 1430:1458 substrates 1449:1458 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 4 30 theme glycans 744:750 arg1 transfer 732:739 transfer 732:739 transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm 732:857 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 0 31 theme Protein-Substrate 27:43 arg1 Interactions 45:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 4 32 theme bacterial 839:847 arg1 periplasm 849:857 the bacterial periplasm 835:857 the bacterial periplasm 835:857 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 5 33 theme distance 1111:1118 arg1 measurements 1120:1131 discrete distance measurements 1102:1131 discrete distance measurements 1102:1131 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 34 theme tag 966:968 arg1 inclusion 912:920 the inclusion 908:920 the inclusion of an encoded N-terminal lanthanide-binding tag (LBT) 908:974 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 0 35 theme Membrane 18:25 arg1 Interactions 45:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Polytopic Membrane Protein-Substrate Interactions 8:56 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 5 36 theme donor 1023:1027 arg1 complex 1029:1035 the luminescent (LBT)-Tb(3+) donor complex 994:1035 the luminescent (LBT)-Tb(3+) donor complex 994:1035 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 1 37 theme essential 133:141 arg1 roles 143:147 essential roles 133:147 essential roles 133:147 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 7 38 theme glycan 1442:1447 arg1 substrates 1449:1458 peptide and glycan substrates 1430:1458 substrates 1449:1458 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 3 39 theme resonance 475:483 arg1 LRET 502:505 LRET 502:505 LRET 502:505 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 39 theme resonance 475:483 arg1 transfer 492:499 luminescence resonance energy transfer 462:499 luminescence resonance energy transfer (LRET) 462:506 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 5 40 theme -Tb 1015:1017 arg1 complex 1029:1035 the luminescent (LBT)-Tb(3+) donor complex 994:1035 the luminescent (LBT)-Tb(3+) donor complex 994:1035 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 2 41 theme Biophysical 251:261 arg1 approaches 263:272 Biophysical approaches 251:272 Biophysical approaches that provide structural information to complement and leverage experimentally determined and computationally predicted structures 251:402 Biophysical approaches that provide structural information to complement and leverage experimentally determined and computationally predicted structures are urgently needed. 26918528 4 42 from Campylobacter 703:715 arg1 PglBs 667:671 the PglBs 663:671 the PglBs from Campylobacter jejuni and Campylobacter lari 663:720 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 3 43 theme energy 485:490 arg1 LRET 502:505 LRET 502:505 LRET 502:505 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 43 theme energy 485:490 arg1 transfer 492:499 luminescence resonance energy transfer 462:499 luminescence resonance energy transfer (LRET) 462:506 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 44 with interactions 530:541 arg1 substrates 625:634 partner substrates 617:634 partner substrates 617:634 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 5 45 theme N-terminal 936:945 arg1 LBT 971:973 LBT 971:973 LBT 971:973 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 45 theme N-terminal 936:945 arg1 tag 966:968 an encoded N-terminal lanthanide-binding tag 925:968 an encoded N-terminal lanthanide-binding tag (LBT) 925:974 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 1 46 theme class 232:236 arg1 analyses 215:222 analyses 215:222 analyses of this class of proteins 215:248 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 0 47 theme Resonance 74:82 arg1 Transfer 91:98 Luminescence Resonance Energy Transfer 61:98 Luminescence Resonance Energy Transfer 61:98 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 7 48 dep related 1546:1552 arg1 uncharacterized 1572:1586 uncharacterized 1572:1586 uncharacterized 1572:1586 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 4 49 from jejuni 692:697 arg1 PglBs 667:671 the PglBs 663:671 the PglBs from Campylobacter jejuni and Campylobacter lari 663:720 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 3 50 theme polytopic 550:558 arg1 transferases 590:601 the polytopic membrane-bound oligosaccharyl transferases 546:601 the polytopic membrane-bound oligosaccharyl transferases (OTases) 546:610 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 50 theme polytopic 550:558 arg1 OTases 604:609 OTases 604:609 OTases 604:609 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 51 theme luminescence 462:473 arg1 LRET 502:505 LRET 502:505 LRET 502:505 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 51 theme luminescence 462:473 arg1 transfer 492:499 luminescence resonance energy transfer 462:499 luminescence resonance energy transfer (LRET) 462:506 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 6 52 theme reported 1302:1309 arg1 structure 1336:1344 the C. lari PglB crystal structure 1311:1344 the previously reported the C. lari PglB crystal structure 1287:1344 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 7 53 theme substrate 1521:1529 arg1 binding 1531:1537 substrate binding 1521:1537 substrate binding to the related, but structurally uncharacterized, eukaryotic OTase 1521:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 4 54 dep Campylobacter 703:715 arg1 lari 717:720 Campylobacter lari 703:720 Campylobacter lari 703:720 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 3 55 theme membrane-bound 560:573 arg1 transferases 590:601 the polytopic membrane-bound oligosaccharyl transferases 546:601 the polytopic membrane-bound oligosaccharyl transferases (OTases) 546:610 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 3 55 theme membrane-bound 560:573 arg1 OTases 604:609 OTases 604:609 OTases 604:609 Herein we present the application of luminescence resonance energy transfer (LRET) for investigating the interactions of the polytopic membrane-bound oligosaccharyl transferases (OTases) with partner substrates. 26918528 5 56 theme discrete 1102:1109 arg1 measurements 1120:1131 discrete distance measurements 1102:1131 discrete distance measurements 1102:1131 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 4 57 link diphosphate-linked 790:807 arg1 substrates 809:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 2 58 theme structural 287:296 arg1 information 298:308 structural information 287:308 structural information to complement 287:322 Biophysical approaches that provide structural information to complement and leverage experimentally determined and computationally predicted structures are urgently needed. 26918528 2 59 theme predicted 383:391 arg1 structures 393:402 experimentally determined and computationally predicted structures 337:402 experimentally determined and computationally predicted structures 337:402 Biophysical approaches that provide structural information to complement and leverage experimentally determined and computationally predicted structures are urgently needed. 26918528 5 60 dep -Tb 1015:1017 arg1 3+ 1019:1020 3+ 1019:1020 3+ 1019:1020 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 7 61 from information 1506:1516 arg1 binding 1531:1537 substrate binding 1521:1537 substrate binding to the related, but structurally uncharacterized, eukaryotic OTase 1521:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 6 62 theme crystal 1328:1334 arg1 structure 1336:1344 the C. lari PglB crystal structure 1311:1344 the previously reported the C. lari PglB crystal structure 1287:1344 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 1 63 theme living 156:161 arg1 systems 163:169 all living systems 152:169 all living systems 152:169 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 4 64 from proteins 823:830 arg1 periplasm 849:857 the bacterial periplasm 835:857 the bacterial periplasm 835:857 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 1 65 theme proteins 241:248 arg1 class 232:236 this class 227:236 this class of proteins 227:248 Integral membrane proteins play essential roles in all living systems; however, major technical hurdles challenge analyses of this class of proteins. 26918528 5 66 theme distance 871:878 arg1 measurements 880:891 LRET-based distance measurements 860:891 LRET-based distance measurements 860:891 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 67 theme membrane 1156:1163 arg1 span 1144:1147 the span 1140:1147 the span of the membrane 1140:1163 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 6 68 theme PglB 1323:1326 arg1 structure 1336:1344 the C. lari PglB crystal structure 1311:1344 the previously reported the C. lari PglB crystal structure 1287:1344 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 4 69 theme undecaprenol 777:788 arg1 substrates 809:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 0 70 theme Energy 84:89 arg1 Transfer 91:98 Luminescence Resonance Energy Transfer 61:98 Luminescence Resonance Energy Transfer 61:98 Probing Polytopic Membrane Protein-Substrate Interactions by Luminescence Resonance Energy Transfer. 26918528 7 71 theme new 1489:1491 arg1 information 1506:1516 new experimental information 1489:1516 new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase 1489:1604 Distance measurements between peptide and glycan substrates and the C. jejuni PglB offer new experimental information on substrate binding to the related, but structurally uncharacterized, eukaryotic OTase. 26918528 5 72 theme lanthanide-binding 947:964 arg1 LBT 971:973 LBT 971:973 LBT 971:973 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 5 72 theme lanthanide-binding 947:964 arg1 tag 966:968 an encoded N-terminal lanthanide-binding tag 925:968 an encoded N-terminal lanthanide-binding tag (LBT) 925:974 LRET-based distance measurements are enabled by the inclusion of an encoded N-terminal lanthanide-binding tag (LBT), and LRET between the luminescent (LBT)-Tb(3+) donor complex and fluorescently labeled peptide and glycan substrates provides discrete distance measurements across the span of the membrane. 26918528 6 73 from C. lari 1225:1231 arg1 measurements 1177:1188 LRET-based measurements 1166:1188 LRET-based measurements of detergent-solubilized PglB from C. lari 1166:1231 LRET-based measurements of detergent-solubilized PglB from C. lari allowed direct comparison with the distances based on the previously reported the C. lari PglB crystal structure, thereby validating the approach in a defined system. 26918528 4 74 theme membrane-associated 757:775 arg1 substrates 809:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 membrane-associated undecaprenol diphosphate-linked substrates 757:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 4 75 from substrates 809:818 arg1 glycans 744:750 glycans 744:750 glycans from membrane-associated undecaprenol diphosphate-linked substrates 744:818 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 26918528 4 75 from substrates 809:818 arg1 transfer 732:739 transfer 732:739 transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm 732:857 Monomeric OTases, such as the PglBs from Campylobacter jejuni and Campylobacter lari, catalyze transfer of glycans from membrane-associated undecaprenol diphosphate-linked substrates to proteins in the bacterial periplasm. 25960101 2 0 theme acid 412:415 arg1 part 426:429 a guluronic acid acceptor part 400:429 a guluronic acid acceptor part 400:429 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 3 1 theme reacting 643:650 arg1 group 660:664 the reacting alcohol group 639:664 the reacting alcohol group 639:664 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 2 2 theme linkages 534:541 arg1 construction 499:510 the fully stereoselective construction 473:510 the fully stereoselective construction of the cis-glycosidic linkages 473:541 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 1 3 theme oligosaccharides 167:182 arg1 synthesis 130:138 The total synthesis 120:138 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), 120:250 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 0 4 theme α-L-guluronic 76:88 arg1 acid 90:93 α-L-guluronic acid 76:93 α-L-guluronic acid 76:93 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 0 5 from reactivity 9:18 arg1 synthesis 33:41 the total synthesis 23:41 the total synthesis of alginate fragments 23:63 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 2 6 theme cis-glycosidic 519:532 arg1 linkages 534:541 the cis-glycosidic linkages 515:541 the cis-glycosidic linkages 515:541 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 7 theme GMGM 302:305 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 3 8 with united 778:783 arg1 efficiency 731:740 the efficiency 727:740 the efficiency with which the building blocks were united 727:783 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 0 9 theme β-D-mannuronic 99:112 arg1 acid 114:117 β-D-mannuronic acid 99:117 β-D-mannuronic acid 99:117 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 2 10 theme mannuronic 437:446 arg1 side 459:462 a mannuronic acid donor side 435:462 a mannuronic acid donor side 435:462 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 11 theme conformational 878:891 arg1 factor 941:946 an all-important factor 924:946 an all-important factor in determining the outcome of a glycosylation reaction 924:1001 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 4 11 theme conformational 878:891 arg1 flexibility 893:903 the conformational flexibility 874:903 the conformational flexibility of the acceptor 874:919 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 0 12 theme Acceptor 0:7 arg1 reactivity 9:18 Acceptor reactivity 0:18 Acceptor reactivity in the total synthesis of alginate fragments 0:63 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 2 13 theme GMGMGMG 323:329 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 13 theme GMGMGMG 323:329 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 14 theme building 376:383 arg1 blocks 385:390 GM building blocks 373:390 GM building blocks 373:390 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 15 theme guluronic 402:410 arg1 part 426:429 a guluronic acid acceptor part 400:429 a guluronic acid acceptor part 400:429 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 16 theme acceptor 841:848 arg1 shape 828:832 the overall shape 816:832 the overall shape of the acceptor 816:848 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 1 17 theme β-D-mannuronic 200:213 arg1 M 221:221 M 221:221 M 221:221 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 1 17 theme β-D-mannuronic 200:213 arg1 acid 215:218 β-D-mannuronic acid 200:218 β-D-mannuronic acid (M) 200:222 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 2 18 theme GM 373:374 arg1 blocks 385:390 GM building blocks 373:390 GM building blocks 373:390 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 19 contain having 393:398 arg2 part 426:429 a guluronic acid acceptor part 400:429 a guluronic acid acceptor part 400:429 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 19 contain having 393:398 arg1 blocks 385:390 GM building blocks 373:390 GM building blocks 373:390 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 20 theme acceptor 912:919 arg1 factor 941:946 an all-important factor 924:946 an all-important factor in determining the outcome of a glycosylation reaction 924:1001 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 4 20 theme acceptor 912:919 arg1 flexibility 893:903 the conformational flexibility 874:903 the conformational flexibility of the acceptor 874:919 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 2 21 theme GMGMGM 315:320 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 21 theme GMGMGM 315:320 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 22 theme all-important 927:939 arg1 factor 941:946 an all-important factor 924:946 an all-important factor in determining the outcome of a glycosylation reaction 924:1001 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 4 22 theme all-important 927:939 arg1 flexibility 893:903 the conformational flexibility 874:903 the conformational flexibility of the acceptor 874:919 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 4 23 theme overall 820:826 arg1 shape 828:832 the overall shape 816:832 the overall shape of the acceptor 816:848 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 4 24 theme reaction 994:1001 arg1 outcome 967:973 the outcome 963:973 the outcome of a glycosylation reaction 963:1001 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 1 25 theme first 272:276 arg1 time 278:281 the first time 268:281 the first time 268:281 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 2 26 theme alginates 343:351 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 26 theme alginates 343:351 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 27 theme GMG 297:299 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 28 theme GMGGMG 336:341 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 0 29 theme total 27:31 arg1 synthesis 33:41 the total synthesis 23:41 the total synthesis of alginate fragments 23:63 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 3 30 theme disaccharide 677:688 arg1 acceptor 690:697 the key disaccharide acceptor 669:697 the key disaccharide acceptor 669:697 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 2 31 theme donor 453:457 arg1 side 459:462 a mannuronic acid donor side 435:462 a mannuronic acid donor side 435:462 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 0 32 theme alginate 46:53 arg1 fragments 55:63 alginate fragments 46:63 alginate fragments 46:63 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 2 33 theme acid 448:451 arg1 side 459:462 a mannuronic acid donor side 435:462 a mannuronic acid donor side 435:462 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 34 theme chiral 791:796 arg1 center 798:803 This chiral center 786:803 This chiral center 786:803 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 0 35 theme fragments 55:63 arg1 synthesis 33:41 the total synthesis 23:41 the total synthesis of alginate fragments 23:63 Acceptor reactivity in the total synthesis of alginate fragments containing α-L-guluronic acid and β-D-mannuronic acid. 25960101 3 36 theme alcohol 652:658 arg1 group 660:664 the reacting alcohol group 639:664 the reacting alcohol group 639:664 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 2 37 theme GMGMG 308:312 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 37 theme GMGMG 308:312 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 38 theme stereoselective 483:497 arg1 construction 499:510 the fully stereoselective construction 473:510 the fully stereoselective construction of the cis-glycosidic linkages 473:541 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 3 39 theme anomeric 593:600 arg1 center 602:607 the reducing-end anomeric center 576:607 the reducing-end anomeric center 576:607 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 1 40 theme total 124:128 arg1 synthesis 130:138 The total synthesis 120:138 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), 120:250 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 3 41 theme key 673:675 arg1 acceptor 690:697 the key disaccharide acceptor 669:697 the key disaccharide acceptor 669:697 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 42 theme center 602:607 arg1 nature 566:571 the nature 562:571 the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, 562:698 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 43 contain had 700:702 arg1 nature 566:571 the nature 562:571 the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, 562:698 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 43 contain had 700:702 arg2 effect 717:722 a tremendous effect 704:722 a tremendous effect 704:722 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 44 theme building 757:764 arg1 united 778:783 united 778:783 united 778:783 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 44 theme building 757:764 arg1 blocks 766:771 the building blocks 753:771 the building blocks 753:771 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 1 45 theme α-L-guluronic 228:240 arg1 G 248:248 G 248:248 G 248:248 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 1 45 theme α-L-guluronic 228:240 arg1 acid 242:245 α-L-guluronic acid 228:245 α-L-guluronic acid (G) 228:249 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 2 46 theme GM 293:294 arg1 GMGMGMG 323:329 GMGMGMG 323:329 GMGMGMG 323:329 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 set 286:288 A set 284:288 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates 284:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 GMGM 302:305 GMGM 302:305 GMGM 302:305 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 GMGMG 308:312 GMGMG 308:312 GMGMG 308:312 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 GM 293:294 GM 293:294 GM 293:294 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 alginates 343:351 GMGGMG alginates 336:351 GMGGMG alginates 336:351 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 GMG 297:299 GMG 297:299 GMG 297:299 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 2 46 theme GM 293:294 arg1 GMGMGM 315:320 GMGMGM 315:320 GMGMGM 315:320 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 4 47 theme glycosylation 980:992 arg1 reaction 994:1001 a glycosylation reaction 978:1001 a glycosylation reaction 978:1001 This chiral center determines the overall shape of the acceptor and it is revealed that the conformational flexibility of the acceptor is an all-important factor in determining the outcome of a glycosylation reaction. 25960101 3 48 theme tremendous 706:715 arg1 effect 717:722 a tremendous effect 704:722 a tremendous effect 704:722 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 3 49 theme reducing-end 580:591 arg1 center 602:607 the reducing-end anomeric center 576:607 the reducing-end anomeric center 576:607 It was found that the nature of the reducing-end anomeric center, which is ten atoms away from the reacting alcohol group in the key disaccharide acceptor, had a tremendous effect on the efficiency with which the building blocks were united. 25960101 1 50 theme mixed-sequence 143:156 arg1 oligosaccharides 167:182 mixed-sequence alginate oligosaccharides 143:182 mixed-sequence alginate oligosaccharides 143:182 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25960101 2 51 theme acceptor 417:424 arg1 part 426:429 a guluronic acid acceptor part 400:429 a guluronic acid acceptor part 400:429 A set of GM, GMG, GMGM, GMGMG, GMGMGM, GMGMGMG, and GMGGMG alginates was assembled using GM building blocks, having a guluronic acid acceptor part and a mannuronic acid donor side to allow the fully stereoselective construction of the cis-glycosidic linkages. 25960101 1 52 theme alginate 158:165 arg1 oligosaccharides 167:182 mixed-sequence alginate oligosaccharides 143:182 mixed-sequence alginate oligosaccharides 143:182 The total synthesis of mixed-sequence alginate oligosaccharides, featuring both β-D-mannuronic acid (M) and α-L-guluronic acid (G), is reported for the first time. 25204387 10 0 theme early-divergent 1960:1974 arg1 plants 1981:1986 early-divergent land plants 1960:1986 early-divergent land plants 1960:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 1 1 theme ancestral 209:217 arg1 CGA 219:221 ancestral CGA 209:221 ancestral CGA 209:221 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 10 2 theme present 1949:1955 arg1 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 2 theme present 1949:1955 arg1 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 3 theme putative 1804:1811 arg1 sequence 1825:1832 a putative CSLA/K-like sequence 1802:1832 a putative CSLA/K-like sequence from a CGA Spirogyra species 1802:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 3 4 theme signalling 458:467 arg1 molecules 469:477 signalling molecules 458:477 signalling molecules 458:477 These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. 25204387 10 5 from species 1752:1758 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 5 from species 1752:1758 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 6 located present 2001:2007 arg1 CGA 2012:2014 CGA 2012:2014 CGA 2012:2014 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 6 located present 2001:2007 arg2 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 6 located present 2001:2007 arg2 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 11 7 theme biosynthetic 2197:2208 arg1 capacity 2210:2217 the their cell wall biosynthetic capacity 2177:2217 the their cell wall biosynthetic capacity 2177:2217 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 12 8 theme land 2338:2341 arg1 walls 2354:2358 land plant cell walls 2338:2358 land plant cell walls 2338:2358 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 6 9 dep available 1033:1041 arg1 so 1044:1045 so 1044:1045 so 1044:1045 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 6 10 theme CGA 1133:1135 arg1 machinery 1120:1128 the biosynthetic machinery 1103:1128 the biosynthetic machinery of CGA 1103:1135 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 5 11 theme cell 780:783 arg1 wall 785:788 the land plant cell wall 765:788 the land plant cell wall 765:788 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 10 12 from orbicularis 1772:1782 arg1 species 1855:1861 a CGA Spirogyra species 1839:1861 a CGA Spirogyra species 1839:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 12 13 theme cell 2349:2352 arg1 walls 2354:2358 land plant cell walls 2338:2358 land plant cell walls 2338:2358 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 6 14 theme machinery 1120:1128 arg1 evolution 1090:1098 the evolution 1086:1098 the evolution of the biosynthetic machinery of CGA 1086:1135 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 3 15 dep cell 515:518 arg1 differentiation 520:534 differentiation 520:534 differentiation 520:534 These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. 25204387 10 16 from present 2001:2007 arg1 CGA 2012:2014 CGA 2012:2014 CGA 2012:2014 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 17 contain contain 879:885 arg2 number 893:898 a low number 887:898 a low number of glycosyl transferases (GTs) 887:929 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 5 17 contain contain 879:885 arg1 genomes 871:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 4 18 theme cell 556:559 arg1 wall 561:564 The cell wall 552:564 The cell wall of land plants 552:579 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 4 18 theme cell 556:559 arg1 composite 607:615 a highly complex fibre composite 584:615 a highly complex fibre composite 584:615 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 10 19 from present 1949:1955 arg1 plants 1981:1986 early-divergent land plants 1960:1986 early-divergent land plants 1960:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 20 theme sequence 1825:1832 arg1 orbicularis 1772:1782 Coleochaete orbicularis 1760:1782 Coleochaete orbicularis 1760:1782 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 20 theme sequence 1825:1832 arg1 fragment 1790:1797 a fragment 1788:1797 a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1788:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 20 theme sequence 1825:1832 arg1 species 1752:1758 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 1 21 theme living 168:173 arg1 algae 126:130 The charophyte green algae 105:130 The charophyte green algae (CGA) 105:136 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 21 theme living 168:173 arg1 relatives 175:183 the closest living relatives 156:183 the closest living relatives to the land plants 156:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 22 theme charophyte 109:118 arg1 CGA 133:135 CGA 133:135 CGA 133:135 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 22 theme charophyte 109:118 arg1 algae 126:130 The charophyte green algae 105:130 The charophyte green algae (CGA) 105:136 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 22 theme charophyte 109:118 arg1 relatives 175:183 the closest living relatives 156:183 the closest living relatives to the land plants 156:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 6 23 theme biosynthetic 1107:1118 arg1 machinery 1120:1128 the biosynthetic machinery 1103:1128 the biosynthetic machinery of CGA 1103:1135 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 10 24 attach present 1949:1955 arg2 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 24 attach present 1949:1955 arg1 plants 1981:1986 early-divergent land plants 1960:1986 early-divergent land plants 1960:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 24 attach present 1949:1955 arg2 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 25 theme land 769:772 arg1 wall 785:788 the land plant cell wall 765:788 the land plant cell wall 765:788 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 11 26 dep CONCLUSIONS 2017:2027 arg1 provide 2041:2047 provide 2041:2047 provide new insights into the evolution of cell walls 2041:2093 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 11 26 dep CONCLUSIONS 2017:2027 arg1 support 2099:2105 support 2099:2105 support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity 2099:2217 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 10 27 theme Spirogyra 1845:1853 arg1 species 1855:1861 a CGA Spirogyra species 1839:1861 a CGA Spirogyra species 1839:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 28 theme chlorophyte 836:846 arg1 genomes 871:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 7 29 theme wall 1243:1246 arg1 GTs 1261:1263 cell wall biosynthesis GTs 1238:1263 cell wall biosynthesis GTs 1238:1263 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 6 30 from number 972:977 arg1 CGA 989:991 CGA 989:991 CGA 989:991 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 6 31 theme transcriptomes 1170:1183 arg1 analysis 1148:1155 an analysis 1145:1155 an analysis of available transcriptomes 1145:1183 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 10 32 theme D 1716:1716 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 32 theme D 1716:1716 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 33 theme prasinophyte 852:863 arg1 algae 865:869 prasinophyte algae 852:869 prasinophyte algae 852:869 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 12 34 from transition 2480:2489 arg1 inherent 2463:2470 inherent 2463:2470 inherent 2463:2470 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 11 35 theme walls 2089:2093 arg1 evolution 2071:2079 the evolution 2067:2079 the evolution of cell walls 2067:2093 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 11 36 theme cell 2187:2190 arg1 wall 2192:2195 their cell wall 2181:2195 the their cell wall biosynthetic capacity 2177:2217 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 12 37 theme selection 2443:2451 arg1 pressures 2453:2461 selection pressures 2443:2461 selection pressures inherent in this transition 2443:2489 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 4 38 theme complex 593:599 arg1 wall 561:564 The cell wall 552:564 The cell wall of land plants 552:579 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 4 38 theme complex 593:599 arg1 composite 607:615 a highly complex fibre composite 584:615 a highly complex fibre composite 584:615 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 10 39 theme cellulose 1692:1700 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 39 theme cellulose 1692:1700 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 4 40 from embedded 713:720 arg1 matrix 727:732 a matrix 725:732 a matrix of pectic polysaccharides 725:758 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 12 41 from inherent 2463:2470 arg1 transition 2480:2489 this transition 2475:2489 this transition 2475:2489 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 12 42 theme inherent 2463:2470 arg1 pressures 2453:2461 selection pressures 2443:2461 selection pressures inherent in this transition 2443:2489 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 0 43 theme green 72:76 arg1 algae 78:82 charophyte green algae 61:82 charophyte green algae 61:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 8 44 theme important 1379:1387 arg1 questions 1402:1410 important evolutionary questions 1379:1410 important evolutionary questions 1379:1410 In addition, gene cloning was employed in two cases to answer important evolutionary questions. 25204387 2 45 theme defining 318:325 arg1 wall 286:289 The cell wall 277:289 The cell wall 277:289 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 2 45 theme defining 318:325 arg1 structure 327:335 a defining structure 316:335 a defining structure that enabled the green algal ancestor to colonize land 316:390 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 5 46 theme glycosyl 903:910 arg1 GTs 926:928 GTs 926:928 GTs 926:928 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 5 46 theme glycosyl 903:910 arg1 transferases 912:923 glycosyl transferases 903:923 glycosyl transferases (GTs) 903:929 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 6 47 theme GTs 982:984 arg1 unknown 1006:1012 unknown 1006:1012 unknown 1006:1012 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 6 47 theme GTs 982:984 arg1 number 972:977 The number 968:977 The number of GTs in CGA 968:991 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 10 48 from species 1855:1861 arg1 species 1752:1758 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 48 from species 1855:1861 arg1 orbicularis 1772:1782 Coleochaete orbicularis 1760:1782 Coleochaete orbicularis 1760:1782 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 48 from species 1855:1861 arg1 fragment 1790:1797 a fragment 1788:1797 a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1788:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 48 from species 1855:1861 arg1 sequence 1825:1832 a putative CSLA/K-like sequence 1802:1832 a putative CSLA/K-like sequence from a CGA Spirogyra species 1802:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 2 49 theme algal 360:364 arg1 ancestor 366:373 the green algal ancestor 350:373 the green algal ancestor 350:373 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 0 50 theme land 13:16 arg1 mechanisms 47:56 land plant cell wall biosynthetic mechanisms 13:56 land plant cell wall biosynthetic mechanisms in charophyte green algae 13:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 9 51 from protein 1660:1666 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 7 52 theme Available 1194:1202 arg1 transcriptomes 1208:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 4 53 theme plants 574:579 arg1 wall 561:564 The cell wall 552:564 The cell wall of land plants 552:579 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 4 53 theme plants 574:579 arg1 composite 607:615 a highly complex fibre composite 584:615 a highly complex fibre composite 584:615 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 0 54 theme cell 24:27 arg1 mechanisms 47:56 land plant cell wall biosynthetic mechanisms 13:56 land plant cell wall biosynthetic mechanisms in charophyte green algae 13:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 4 55 theme polysaccharides 744:758 arg1 matrix 727:732 a matrix 725:732 a matrix of pectic polysaccharides 725:758 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 2 56 theme cell 281:284 arg1 structure 327:335 a defining structure 316:335 a defining structure that enabled the green algal ancestor to colonize land 316:390 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 2 56 theme cell 281:284 arg1 wall 286:289 The cell wall 277:289 The cell wall 277:289 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 0 57 theme biosynthetic 34:45 arg1 mechanisms 47:56 land plant cell wall biosynthetic mechanisms 13:56 land plant cell wall biosynthetic mechanisms in charophyte green algae 13:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 1 58 theme land 257:260 arg1 lineage 268:274 the land plant lineage 253:274 the land plant lineage 253:274 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 9 59 theme core 1498:1501 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 12 60 theme cell 2282:2285 arg1 evolution 2292:2300 plant cell wall evolution 2276:2300 plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land 2276:2398 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 10 61 theme CGA 1748:1750 arg1 orbicularis 1772:1782 Coleochaete orbicularis 1760:1782 Coleochaete orbicularis 1760:1782 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 61 theme CGA 1748:1750 arg1 fragment 1790:1797 a fragment 1788:1797 a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1788:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 61 theme CGA 1748:1750 arg1 species 1752:1758 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 4 62 theme embedded 713:720 arg1 xyloglucan 701:710 xyloglucan 701:710 xyloglucan 701:710 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 3 63 theme cell 399:402 arg1 walls 404:408 These cell walls 393:408 These cell walls 393:408 These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. 25204387 9 64 from pectin 1624:1629 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 65 theme wall 1508:1511 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 10 66 theme cellulose 1918:1926 arg1 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 66 theme cellulose 1918:1926 arg1 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 6 67 from CGA 989:991 arg1 unknown 1006:1012 unknown 1006:1012 unknown 1006:1012 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 6 67 from CGA 989:991 arg1 number 972:977 The number 968:977 The number of GTs in CGA 968:991 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 10 68 theme land 1976:1979 arg1 plants 1981:1986 early-divergent land plants 1960:1986 early-divergent land plants 1960:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 6 69 theme available 1160:1168 arg1 transcriptomes 1170:1183 available transcriptomes 1160:1183 available transcriptomes 1160:1183 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 9 70 theme evolutionary 1540:1551 arg1 origins 1553:1559 their evolutionary origins 1534:1559 their evolutionary origins 1534:1559 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 10 71 theme Coleochaete 1760:1770 arg1 orbicularis 1772:1782 Coleochaete orbicularis 1760:1782 Coleochaete orbicularis 1760:1782 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 71 theme Coleochaete 1760:1770 arg1 species 1752:1758 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 9 72 theme important 1488:1496 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 12 73 theme plant 2343:2347 arg1 walls 2354:2358 land plant cell walls 2338:2358 land plant cell walls 2338:2358 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 6 74 from GTs 982:984 arg1 CGA 989:991 CGA 989:991 CGA 989:991 The number of GTs in CGA is currently unknown, as no genomes are available, so this study sought to give insight into the evolution of the biosynthetic machinery of CGA through an analysis of available transcriptomes. 25204387 5 75 theme plant 774:778 arg1 wall 785:788 the land plant cell wall 765:788 the land plant cell wall 765:788 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 12 76 theme walls 2354:2358 arg1 features 2326:2333 some features 2321:2333 some features of land plant cell walls 2321:2358 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 4 77 theme land 569:572 arg1 plants 574:579 land plants 569:579 land plants 569:579 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 7 78 theme land 1304:1307 arg1 plants 1309:1314 land plants 1304:1314 land plants 1304:1314 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 3 79 theme molecules 469:477 arg1 source 448:453 a source 446:453 a source of signalling molecules 446:477 These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. 25204387 1 80 theme closest 160:166 arg1 algae 126:130 The charophyte green algae 105:130 The charophyte green algae (CGA) 105:136 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 80 theme closest 160:166 arg1 relatives 175:183 the closest living relatives 156:183 the closest living relatives to the land plants 156:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 8 81 theme gene 1330:1333 arg1 cloning 1335:1341 gene cloning 1330:1341 gene cloning 1330:1341 In addition, gene cloning was employed in two cases to answer important evolutionary questions. 25204387 4 82 theme fibre 601:605 arg1 wall 561:564 The cell wall 552:564 The cell wall of land plants 552:579 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 4 82 theme fibre 601:605 arg1 composite 607:615 a highly complex fibre composite 584:615 a highly complex fibre composite 584:615 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 9 83 contain have 1529:1532 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 83 contain have 1529:1532 arg2 cellulose 1583:1591 cellulose 1583:1591 cellulose 1583:1591 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 83 contain have 1529:1532 arg2 protein 1660:1666 arabino-galactan protein 1643:1666 arabino-galactan protein 1643:1666 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 83 contain have 1529:1532 arg1 many 1471:1474 many 1471:1474 many 1471:1474 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 83 contain have 1529:1532 arg2 origins 1553:1559 their evolutionary origins 1534:1559 their evolutionary origins 1534:1559 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 10 84 theme CSLA/K-like 1813:1823 arg1 sequence 1825:1832 a putative CSLA/K-like sequence 1802:1832 a putative CSLA/K-like sequence from a CGA Spirogyra species 1802:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 85 theme putative 1683:1690 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 85 theme putative 1683:1690 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 86 from fragment 1790:1797 arg1 species 1855:1861 a CGA Spirogyra species 1839:1861 a CGA Spirogyra species 1839:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 9 87 from origins 1553:1559 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 3 88 theme developmental 492:504 arg1 cues 506:509 developmental cues 492:509 developmental cues for cell differentiation and elongation 492:549 These cell walls provide support and protection, are a source of signalling molecules, and provide developmental cues for cell differentiation and elongation. 25204387 5 89 theme early-divergent 820:834 arg1 chlorophyte 836:846 early-divergent chlorophyte 820:846 early-divergent chlorophyte 820:846 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 10 90 theme CGA 1841:1843 arg1 species 1855:1861 a CGA Spirogyra species 1839:1861 a CGA Spirogyra species 1839:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 91 theme transferases 912:923 arg1 number 893:898 a low number 887:898 a low number of glycosyl transferases (GTs) 887:929 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 10 92 theme family 1718:1723 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 92 theme family 1718:1723 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 4 93 from matrix 727:732 arg1 embedded 713:720 embedded 713:720 embedded 713:720 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 1 94 theme green 120:124 arg1 CGA 133:135 CGA 133:135 CGA 133:135 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 94 theme green 120:124 arg1 algae 126:130 The charophyte green algae 105:130 The charophyte green algae (CGA) 105:136 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 1 94 theme green 120:124 arg1 relatives 175:183 the closest living relatives 156:183 the closest living relatives to the land plants 156:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 9 95 from xylan 1614:1618 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 7 96 theme biosynthesis 1248:1259 arg1 GTs 1261:1263 cell wall biosynthesis GTs 1238:1263 cell wall biosynthesis GTs 1238:1263 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 12 97 theme pressures 2453:2461 arg1 result 2433:2438 a result 2431:2438 a result of selection pressures inherent in this transition 2431:2489 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 10 98 from CGA 2012:2014 arg1 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 98 from CGA 2012:2014 arg1 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 0 99 theme charophyte 61:70 arg1 algae 78:82 charophyte green algae 61:82 charophyte green algae 61:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 10 100 theme synthase-like 1702:1714 arg1 CSLDs 1732:1736 CSLDs 1732:1736 CSLDs 1732:1736 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 100 theme synthase-like 1702:1714 arg1 genes 1725:1729 two putative cellulose synthase-like D family genes 1679:1729 two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1679:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 9 101 from xyloglucan 1602:1611 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 7 102 theme cell 1238:1241 arg1 GTs 1261:1263 cell wall biosynthesis GTs 1238:1263 cell wall biosynthesis GTs 1238:1263 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 9 103 theme KEY 1413:1415 arg1 RESULTS 1417:1423 KEY RESULTS 1413:1423 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 1413:1667 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 104 from mannan 1594:1599 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 11 105 theme cell 2084:2087 arg1 walls 2089:2093 cell walls 2084:2093 cell walls 2084:2093 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 8 106 theme evolutionary 1389:1400 arg1 questions 1402:1410 important evolutionary questions 1379:1410 important evolutionary questions 1379:1410 In addition, gene cloning was employed in two cases to answer important evolutionary questions. 25204387 1 107 dep BACKGROUND 85:94 arg1 thought 142:148 thought 142:148 are thought to be the closest living relatives to the land plants 138:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 5 108 theme low 889:891 arg1 number 893:898 a low number 887:898 a low number of glycosyl transferases (GTs) 887:929 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 9 109 from cellulose 1583:1591 arg1 CGA 1568:1570 the CGA 1564:1570 the CGA 1564:1570 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 110 theme arabino-galactan 1643:1658 arg1 protein 1660:1666 arabino-galactan protein 1643:1666 arabino-galactan protein 1643:1666 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 0 111 from mechanisms 47:56 arg1 algae 78:82 charophyte green algae 61:82 charophyte green algae 61:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 11 112 theme capacity 2210:2217 arg1 virtue 2167:2172 virtue 2167:2172 virtue of the their cell wall biosynthetic capacity 2167:2217 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 10 113 theme first 1890:1894 arg1 evidence 1896:1903 the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA 1886:2014 the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA 1886:2014 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 114 attach present 2001:2007 arg1 CGA 2012:2014 CGA 2012:2014 CGA 2012:2014 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 114 attach present 2001:2007 arg2 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 114 attach present 2001:2007 arg2 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 12 115 dep evolution 2292:2300 arg1 imply 2310:2314 imply 2310:2314 imply that some features of land plant cell walls evolved prior to the transition to land 2310:2398 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 5 116 theme algae 865:869 arg1 genomes 871:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 early-divergent chlorophyte and prasinophyte algae genomes 820:877 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 7 117 theme CGA 1204:1206 arg1 transcriptomes 1208:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 11 118 theme new 2049:2051 arg1 insights 2053:2060 new insights 2049:2060 new insights into the evolution of cell walls 2049:2093 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 9 119 dep RESULTS 1417:1423 arg1 obtained 1446:1453 obtained 1446:1453 was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein 1442:1666 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 0 120 theme plant 18:22 arg1 mechanisms 47:56 land plant cell wall biosynthetic mechanisms 13:56 land plant cell wall biosynthetic mechanisms in charophyte green algae 13:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 7 121 theme METHODS 1186:1192 arg1 transcriptomes 1208:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes 1186:1221 METHODS Available CGA transcriptomes were mined for cell wall biosynthesis GTs and compared with GTs characterized in land plants. 25204387 2 122 theme green 354:358 arg1 ancestor 366:373 the green algal ancestor 350:373 the green algal ancestor 350:373 The cell wall has been suggested to be a defining structure that enabled the green algal ancestor to colonize land. 25204387 4 123 theme non-cellulosic 661:674 arg1 polysaccharides 676:690 non-cellulosic polysaccharides 661:690 non-cellulosic polysaccharides 661:690 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 4 123 theme non-cellulosic 661:674 arg1 xyloglucan 701:710 xyloglucan 701:710 xyloglucan 701:710 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 0 124 theme wall 29:32 arg1 mechanisms 47:56 land plant cell wall biosynthetic mechanisms 13:56 land plant cell wall biosynthetic mechanisms in charophyte green algae 13:82 Evidence for land plant cell wall biosynthetic mechanisms in charophyte green algae. 25204387 12 125 theme plant 2276:2280 arg1 evolution 2292:2300 plant cell wall evolution 2276:2300 plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land 2276:2398 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 10 126 from plants 1981:1986 arg1 present 1949:1955 present 1949:1955 present 1949:1955 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 5 127 theme land 938:941 arg1 plants 943:948 land plants 938:948 land plants 938:948 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 5 128 contain contain 950:956 arg2 hundreds 958:965 hundreds 958:965 hundreds 958:965 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 5 128 contain contain 950:956 arg1 plants 943:948 land plants 938:948 land plants 938:948 How the land plant cell wall evolved is currently unknown: early-divergent chlorophyte and prasinophyte algae genomes contain a low number of glycosyl transferases (GTs), while land plants contain hundreds. 25204387 12 129 theme wall 2287:2290 arg1 evolution 2292:2300 plant cell wall evolution 2276:2300 plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land 2276:2398 These findings are highly significant for understanding plant cell wall evolution as they imply that some features of land plant cell walls evolved prior to the transition to land, rather than having evolved as a result of selection pressures inherent in this transition. 25204387 9 130 theme Genetic 1425:1431 arg1 evidence 1433:1440 Genetic evidence 1425:1440 Genetic evidence 1425:1440 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 4 131 theme pectic 737:742 arg1 polysaccharides 744:758 pectic polysaccharides 737:758 pectic polysaccharides 737:758 The cell wall of land plants is a highly complex fibre composite, characterized by cellulose cross-linked by non-cellulosic polysaccharides, such as xyloglucan, embedded in a matrix of pectic polysaccharides. 25204387 1 132 theme plant 262:266 arg1 lineage 268:274 the land plant lineage 253:274 the land plant lineage 253:274 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 9 133 theme cell 1503:1506 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 10 134 theme synthase/-like 1928:1941 arg1 genes 1943:1947 all the cellulose synthase/-like genes 1910:1947 all the cellulose synthase/-like genes present in early-divergent land plants 1910:1986 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 134 theme synthase/-like 1928:1941 arg1 present 2001:2007 present 2001:2007 present 2001:2007 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 1 135 theme land 192:195 arg1 plants 197:202 the land plants 188:202 the land plants 188:202 BACKGROUND AND AIMS The charophyte green algae (CGA) are thought to be the closest living relatives to the land plants, and ancestral CGA were unique in giving rise to the land plant lineage. 25204387 9 136 theme polysaccharides 1513:1527 arg1 polysaccharides 1513:1527 the most important core cell wall polysaccharides 1479:1527 the most important core cell wall polysaccharides 1479:1527 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 9 136 theme polysaccharides 1513:1527 arg1 many 1471:1474 many 1471:1474 many 1471:1474 KEY RESULTS Genetic evidence was obtained indicating that many of the most important core cell wall polysaccharides have their evolutionary origins in the CGA, including cellulose, mannan, xyloglucan, xylan and pectin, as well as arabino-galactan protein. 25204387 11 137 theme wall 2192:2195 arg1 capacity 2210:2217 the their cell wall biosynthetic capacity 2177:2217 the their cell wall biosynthetic capacity 2177:2217 CONCLUSIONS The results provide new insights into the evolution of cell walls and support the notion that the CGA were pre-adapted to life on land by virtue of the their cell wall biosynthetic capacity. 25204387 10 138 dep species 1752:1758 arg1 orbicularis 1772:1782 Coleochaete orbicularis 1760:1782 Coleochaete orbicularis 1760:1782 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 138 dep species 1752:1758 arg1 fragment 1790:1797 a fragment 1788:1797 a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1788:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 25204387 10 138 dep species 1752:1758 arg1 species 1752:1758 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species 1744:1861 Moreover, two putative cellulose synthase-like D family genes (CSLDs) from the CGA species Coleochaete orbicularis and a fragment of a putative CSLA/K-like sequence from a CGA Spirogyra species were cloned, providing the first evidence that all the cellulose synthase/-like genes present in early-divergent land plants were already present in CGA. 26959529 8 0 theme human 1070:1074 arg1 samples 1081:1087 human milk samples 1070:1087 human milk samples 1070:1087 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. 26959529 5 1 theme free 629:632 arg1 oligosaccharides 634:649 free oligosaccharides 629:649 free oligosaccharides 629:649 Quantitation profiles of free oligosaccharides and N-glycans were reported. 26959529 2 2 from component 301:309 arg1 oligosaccharides 319:334 milk oligosaccharides 314:334 milk oligosaccharides 314:334 The main component in milk oligosaccharides is free oligosaccharides. 26959529 9 3 dep sialylated 1183:1192 arg1 fucosylated 1217:1227 fucosylated 1217:1227 fucosylated 1217:1227 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 11 4 theme isomeric 1345:1352 arg1 glycans 1354:1360 the isomeric glycans 1341:1360 the isomeric glycans in milk samples 1341:1376 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 6 5 theme isomeric 774:781 arg1 consideration 783:795 isomeric consideration 774:795 isomeric consideration 774:795 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 1 6 theme pathogen 257:264 arg1 binding 266:272 pathogen binding 257:272 pathogen binding to the host cell 257:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 15 7 theme several 1887:1893 arg1 isomers 1895:1901 several isomers 1887:1901 several isomers 1887:1901 This feature resulted in the identification of several isomers. 26959529 6 8 theme milk 752:755 arg1 samples 757:763 human milk samples 746:763 human milk samples (without isomeric consideration) 746:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 0 9 theme milk 108:111 arg1 samples 113:119 goat milk samples 103:119 goat milk samples 103:119 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 9 10 theme whey 1157:1160 arg1 proteins 1162:1169 whey proteins 1157:1169 whey proteins 1157:1169 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 12 11 theme Gal-GlcNAc-Gal-Glc-Fuc 1538:1559 arg1 isomers 1561:1567 Gal-GlcNAc-Gal-Glc-Fuc isomers 1538:1567 Gal-GlcNAc-Gal-Glc-Fuc isomers 1538:1567 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 13 12 theme MS/MS 1694:1698 arg1 interpretation 1676:1689 the interpretation 1672:1689 the interpretation of MS/MS 1672:1698 Permethylation of the glycan structures facilitated the interpretation of MS/MS. 26959529 12 13 theme Gal-GlcNAc- 1509:1519 arg1 -Gal-Glc 1525:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 14 14 theme glycans 1831:1837 arg1 spectra 1806:1812 the tandem mass spectra 1790:1812 the tandem mass spectra of permethylated glycans 1790:1837 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 12 15 theme Fuc 1521:1523 arg1 -Gal-Glc 1525:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 11 16 theme porous 1397:1402 arg1 LC 1421:1422 porous graphitic carbon LC 1397:1422 porous graphitic carbon LC 1397:1422 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 12 17 theme human 1477:1481 arg1 milk 1483:1486 human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers 1477:1567 milk 1483:1486 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 11 18 theme carbon 1414:1419 arg1 LC 1421:1422 porous graphitic carbon LC 1397:1422 porous graphitic carbon LC 1397:1422 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 14 19 theme mass 1801:1804 arg1 spectra 1806:1812 the tandem mass spectra 1790:1812 the tandem mass spectra of permethylated glycans 1790:1837 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 12 20 theme free 1488:1491 arg1 -Gal-Glc 1525:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 13 21 theme structures 1649:1658 arg1 Permethylation 1620:1633 Permethylation 1620:1633 Permethylation of the glycan structures 1620:1658 Permethylation of the glycan structures facilitated the interpretation of MS/MS. 26959529 8 22 theme N-glycan 949:956 arg1 structures 958:967 Totally 58, 21, and 43 N-glycan structures 926:967 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) 926:1000 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. 26959529 7 23 theme milk 912:915 arg1 samples 917:923 the other two milk samples 898:923 the other two milk samples 898:923 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 9 24 theme milk 1111:1114 arg1 oligosaccharides 1121:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 7 25 theme other 902:906 arg1 samples 917:923 the other two milk samples 898:923 the other two milk samples 898:923 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 12 26 theme carbon 1605:1610 arg1 column 1612:1617 porous graphitic carbon column 1588:1617 porous graphitic carbon column 1588:1617 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 3 27 from proteins 372:379 arg1 milk 384:387 milk 384:387 milk 384:387 Since the proteins in milk are highly glycosylated, N-glycans in milk also play an import role. 26959529 8 28 theme isomeric 978:985 arg1 consideration 987:999 isomeric consideration 978:999 isomeric consideration 978:999 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. 26959529 15 29 theme isomers 1895:1901 arg1 identification 1869:1882 the identification 1865:1882 the identification of several isomers 1865:1901 This feature resulted in the identification of several isomers. 26959529 0 30 theme LC-MS/MS 0:7 arg1 analysis 9:16 LC-MS/MS analysis 0:16 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans 0:69 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 1 31 theme binding 266:272 arg1 inhibition 243:252 inhibition 243:252 inhibition of pathogen binding to the host cell 243:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 7 32 theme oligosaccharides 865:880 arg1 structures 882:891 more complex free oligosaccharides structures 847:891 more complex free oligosaccharides structures 847:891 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 10 33 theme human 1239:1243 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 12 34 theme porous 1588:1593 arg1 column 1612:1617 porous graphitic carbon column 1588:1617 porous graphitic carbon column 1588:1617 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 10 35 gly fucosylated 1322:1332 arg1 N-glycans 1276:1284 N-glycans 1276:1284 N-glycans 1276:1284 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 10 35 gly fucosylated 1322:1332 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 10 36 theme Goat 1230:1233 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 0 37 theme free 35:38 arg1 oligosaccharides 40:55 permethylated free oligosaccharides 21:55 permethylated free oligosaccharides 21:55 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 8 38 theme whey 1023:1026 arg1 proteins 1028:1035 whey proteins 1023:1035 whey proteins extracted from bovine, goat, and human milk samples, respectively 1023:1101 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. 26959529 10 39 from proteins 1296:1303 arg1 N-glycans 1276:1284 N-glycans 1276:1284 N-glycans 1276:1284 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 10 39 from proteins 1296:1303 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 14 40 theme glycosidic 1736:1745 arg1 cleavage 1752:1759 glycosidic bond cleavage 1736:1759 glycosidic bond cleavage 1736:1759 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 4 41 theme free 507:510 arg1 oligosaccharides 512:527 the permethylated free oligosaccharides 489:527 the permethylated free oligosaccharides 489:527 In this study, we investigated the permethylated free oligosaccharides and N-glycans extracted from bovine, goat, and human milks using LC-MS/MS. 26959529 10 42 theme free 1250:1253 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 7 43 theme free 860:863 arg1 structures 882:891 more complex free oligosaccharides structures 847:891 more complex free oligosaccharides structures 847:891 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 1 44 theme significant 203:213 arg1 biofunctions 222:233 significant immune biofunctions 203:233 significant immune biofunctions such as inhibition of pathogen binding to the host cell 203:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 1 44 theme significant 203:213 arg1 inhibition 243:252 inhibition 243:252 inhibition of pathogen binding to the host cell 243:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 3 45 from N-glycans 414:422 arg1 milk 427:430 milk 427:430 milk 427:430 Since the proteins in milk are highly glycosylated, N-glycans in milk also play an import role. 26959529 6 46 theme oligosaccharides 699:714 arg1 number 684:689 The number 680:689 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) 680:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 6 46 theme oligosaccharides 699:714 arg1 8 807:807 8 807:807 8 807:807 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 2 47 theme free 339:342 arg1 component 301:309 The main component 292:309 The main component in milk oligosaccharides 292:334 The main component in milk oligosaccharides is free oligosaccharides. 26959529 2 47 theme free 339:342 arg1 oligosaccharides 344:359 free oligosaccharides 339:359 free oligosaccharides 339:359 The main component in milk oligosaccharides is free oligosaccharides. 26959529 5 48 theme oligosaccharides 634:649 arg1 profiles 617:624 Quantitation profiles 604:624 Quantitation profiles of free oligosaccharides and N-glycans 604:663 Quantitation profiles of free oligosaccharides and N-glycans were reported. 26959529 9 49 theme Bovine 1104:1109 arg1 oligosaccharides 1121:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 8 50 theme milk 1076:1079 arg1 samples 1081:1087 human milk samples 1070:1087 human milk samples 1070:1087 Totally 58, 21, and 43 N-glycan structures (without isomeric consideration) were associated with whey proteins extracted from bovine, goat, and human milk samples, respectively. 26959529 9 51 theme free 1116:1119 arg1 oligosaccharides 1121:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 9 52 from proteins 1162:1169 arg1 N-glycans 1142:1150 N-glycans 1142:1150 N-glycans from whey proteins 1142:1169 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 9 52 from proteins 1162:1169 arg1 oligosaccharides 1121:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 5 53 theme Quantitation 604:615 arg1 profiles 617:624 Quantitation profiles 604:624 Quantitation profiles of free oligosaccharides and N-glycans 604:663 Quantitation profiles of free oligosaccharides and N-glycans were reported. 26959529 2 54 theme main 296:299 arg1 component 301:309 The main component 292:309 The main component in milk oligosaccharides 292:334 The main component in milk oligosaccharides is free oligosaccharides. 26959529 2 54 theme main 296:299 arg1 oligosaccharides 344:359 free oligosaccharides 339:359 free oligosaccharides 339:359 The main component in milk oligosaccharides is free oligosaccharides. 26959529 6 55 theme human 746:750 arg1 samples 757:763 human milk samples 746:763 human milk samples (without isomeric consideration) 746:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 12 56 theme isomers 1561:1567 arg1 separation 1463:1472 separation 1463:1472 separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers 1463:1567 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 9 57 gly sialylated 1183:1192 arg1 N-glycans 1142:1150 N-glycans 1142:1150 N-glycans from whey proteins 1142:1169 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 9 57 gly sialylated 1183:1192 arg1 oligosaccharides 1121:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides 1104:1136 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 5 58 theme N-glycans 655:663 arg1 profiles 617:624 Quantitation profiles 604:624 Quantitation profiles of free oligosaccharides and N-glycans 604:663 Quantitation profiles of free oligosaccharides and N-glycans were reported. 26959529 14 59 theme permethylated 1817:1829 arg1 glycans 1831:1837 permethylated glycans 1817:1837 permethylated glycans 1817:1837 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 12 60 theme milk 1483:1486 arg1 separation 1463:1472 separation 1463:1472 separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers 1463:1567 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 11 61 theme graphitic 1404:1412 arg1 LC 1421:1422 porous graphitic carbon LC 1397:1422 porous graphitic carbon LC 1397:1422 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 12 62 theme oligosaccharide 1493:1507 arg1 -Gal-Glc 1525:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 3 63 gly glycosylated 400:411 arg1 proteins 372:379 the proteins 368:379 the proteins in milk 368:387 Since the proteins in milk are highly glycosylated, N-glycans in milk also play an import role. 26959529 14 64 theme tandem 1794:1799 arg1 spectra 1806:1812 the tandem mass spectra 1790:1812 the tandem mass spectra of permethylated glycans 1790:1837 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 11 65 theme elevated 1427:1434 arg1 temperatures 1436:1447 elevated temperatures 1427:1447 elevated temperatures 1427:1447 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 6 66 theme free 694:697 arg1 oligosaccharides 699:714 free oligosaccharides 694:714 free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) 694:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 0 67 theme goat 103:106 arg1 samples 113:119 goat milk samples 103:119 goat milk samples 103:119 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 11 68 theme milk 1365:1368 arg1 samples 1370:1376 milk samples 1365:1376 milk samples 1365:1376 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 12 69 theme graphitic 1595:1603 arg1 column 1612:1617 porous graphitic carbon column 1588:1617 porous graphitic carbon column 1588:1617 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 7 70 contain had 843:845 arg1 milk 838:841 Human milk 832:841 Human milk 832:841 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 7 70 contain had 843:845 arg2 structures 882:891 more complex free oligosaccharides structures 847:891 more complex free oligosaccharides structures 847:891 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 14 71 theme internal 1714:1721 arg1 cleavage 1723:1730 internal cleavage 1714:1730 internal cleavage 1714:1730 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 1 72 from Oligosaccharides 122:137 arg1 milk 142:145 milk 142:145 milk 142:145 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 13 73 theme glycan 1642:1647 arg1 structures 1649:1658 the glycan structures 1638:1658 the glycan structures 1638:1658 Permethylation of the glycan structures facilitated the interpretation of MS/MS. 26959529 10 74 theme milk 1245:1248 arg1 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides 1230:1270 oligosaccharides 1255:1270 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 0 75 theme permethylated 21:33 arg1 oligosaccharides 40:55 permethylated free oligosaccharides 21:55 permethylated free oligosaccharides 21:55 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 6 76 located observed 716:723 arg1 samples 757:763 human milk samples 746:763 human milk samples (without isomeric consideration) 746:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 6 76 located observed 716:723 arg2 oligosaccharides 699:714 free oligosaccharides 694:714 free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) 694:796 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 6 76 located observed 716:723 arg1 bovine 728:733 bovine 728:733 bovine 728:733 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 6 76 located observed 716:723 arg1 goat 736:739 goat 736:739 goat 736:739 The number of free oligosaccharides observed in bovine, goat, and human milk samples (without isomeric consideration) were 11, 8, and 11, respectively. 26959529 9 77 dep lesser 1203:1208 arg1 extend 1210:1215 extend 1210:1215 extend 1210:1215 Bovine milk free oligosaccharides and N-glycans from whey proteins were highly sialylated and to a lesser extend fucosylated. 26959529 3 78 theme import 445:450 arg1 role 452:455 an import role 442:455 an import role 442:455 Since the proteins in milk are highly glycosylated, N-glycans in milk also play an import role. 26959529 0 79 theme oligosaccharides 40:55 arg1 analysis 9:16 LC-MS/MS analysis 0:16 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans 0:69 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 1 80 theme host 281:284 arg1 cell 286:289 the host cell 277:289 the host cell 277:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 4 81 theme permethylated 493:505 arg1 oligosaccharides 512:527 the permethylated free oligosaccharides 489:527 the permethylated free oligosaccharides 489:527 In this study, we investigated the permethylated free oligosaccharides and N-glycans extracted from bovine, goat, and human milks using LC-MS/MS. 26959529 0 82 theme N-glycans 61:69 arg1 analysis 9:16 LC-MS/MS analysis 0:16 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans 0:69 LC-MS/MS analysis of permethylated free oligosaccharides and N-glycans derived from human, bovine, and goat milk samples. 26959529 14 83 theme bond 1747:1750 arg1 cleavage 1752:1759 glycosidic bond cleavage 1736:1759 glycosidic bond cleavage 1736:1759 For example, internal cleavage and glycosidic bond cleavage are readily distinguished in the tandem mass spectra of permethylated glycans. 26959529 1 84 contain have 198:201 arg2 inhibition 243:252 inhibition 243:252 inhibition of pathogen binding to the host cell 243:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 1 84 contain have 198:201 arg1 Oligosaccharides 122:137 Oligosaccharides 122:137 Oligosaccharides in milk 122:145 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 1 84 contain have 198:201 arg2 biofunctions 222:233 significant immune biofunctions 203:233 significant immune biofunctions such as inhibition of pathogen binding to the host cell 203:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 2 85 theme milk 314:317 arg1 oligosaccharides 319:334 milk oligosaccharides 314:334 milk oligosaccharides 314:334 The main component in milk oligosaccharides is free oligosaccharides. 26959529 7 86 theme complex 852:858 arg1 structures 882:891 more complex free oligosaccharides structures 847:891 more complex free oligosaccharides structures 847:891 Human milk had more complex free oligosaccharides structures than the other two milk samples. 26959529 11 87 from glycans 1354:1360 arg1 samples 1370:1376 milk samples 1365:1376 milk samples 1365:1376 Also, the isomeric glycans in milk samples were determined by porous graphitic carbon LC at elevated temperatures. 26959529 4 88 theme human 576:580 arg1 milks 582:586 human milks 576:586 human milks using LC-MS/MS 576:601 In this study, we investigated the permethylated free oligosaccharides and N-glycans extracted from bovine, goat, and human milks using LC-MS/MS. 26959529 12 89 dep milk 1483:1486 arg1 -Gal-Glc 1525:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc 1488:1532 For example, separation of human milk free oligosaccharide Gal-GlcNAc-(Fuc)-Gal-Glc and Gal-GlcNAc-Gal-Glc-Fuc isomers was achieved using porous graphitic carbon column. 26959529 1 90 theme immune 215:220 arg1 biofunctions 222:233 significant immune biofunctions 203:233 significant immune biofunctions such as inhibition of pathogen binding to the host cell 203:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 1 90 theme immune 215:220 arg1 inhibition 243:252 inhibition 243:252 inhibition of pathogen binding to the host cell 243:289 Oligosaccharides in milk not only provide nutrition to the infants but also have significant immune biofunctions such as inhibition of pathogen binding to the host cell. 26959529 10 91 theme whey 1291:1294 arg1 proteins 1296:1303 whey proteins 1291:1303 whey proteins 1291:1303 Goat and human milk free oligosaccharides and N-glycans from whey proteins were both highly fucosylated. 26959529 7 92 theme Human 832:836 arg1 milk 838:841 Human milk 832:841 Human milk 832:841 Human milk had more complex free oligosaccharides structures than the other two milk samples. 29255015 0 0 theme crystallographic 86:101 arg1 approaches 139:148 crystallographic and carbohydrate molecular modeling approaches 86:148 crystallographic and carbohydrate molecular modeling approaches 86:148 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 5 1 theme molecule 805:812 arg1 preference 781:790 the conformational preference 762:790 the conformational preference of the whole molecule, alone or within the crystallographic environment 762:862 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 3 2 gly glycoprotein 413:424 arg1 glycoprotein 413:424 the complete glycoprotein 400:424 the complete glycoprotein 400:424 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 1 3 theme major 170:174 arg1 Vp54 192:195 the major capsid protein (Vp54) 166:196 the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) 166:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 6 4 theme glycosylated 977:988 arg1 structure 990:998 the complete glycosylated structure 964:998 the complete glycosylated structure of a glycoprotein 964:1016 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 1 5 theme capsid 176:181 arg1 Vp54 192:195 the major capsid protein (Vp54) 166:196 the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) 166:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 5 6 theme Molecular 668:676 arg1 modeling 678:685 Molecular modeling 668:685 Molecular modeling 668:685 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 0 7 theme carbohydrate 107:118 arg1 approaches 139:148 crystallographic and carbohydrate molecular modeling approaches 86:148 crystallographic and carbohydrate molecular modeling approaches 86:148 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 6 8 theme complete 968:975 arg1 structure 990:998 the complete glycosylated structure 964:998 the complete glycosylated structure of a glycoprotein 964:1016 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 6 9 gly glycosylated 977:988 arg1 structure 990:998 the complete glycosylated structure 964:998 the complete glycosylated structure of a glycoprotein 964:1016 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 5 10 theme conformational 766:779 arg1 preference 781:790 the conformational preference 762:790 the conformational preference of the whole molecule, alone or within the crystallographic environment 762:862 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 2 11 theme Vp54 352:355 arg1 structure 363:371 the previously reported Vp54 X-ray structure 328:371 the previously reported Vp54 X-ray structure 328:371 This prompted a reexamination of the previously reported Vp54 X-ray structure. 29255015 0 12 theme modeling 130:137 arg1 approaches 139:148 crystallographic and carbohydrate molecular modeling approaches 86:148 crystallographic and carbohydrate molecular modeling approaches 86:148 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 3 13 theme molecular 479:487 arg1 modeling 489:496 molecular modeling 479:496 molecular modeling 479:496 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 1 14 theme protein 183:189 arg1 Vp54 192:195 the major capsid protein (Vp54) 166:196 the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) 166:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 0 15 theme molecular 120:128 arg1 approaches 139:148 crystallographic and carbohydrate molecular modeling approaches 86:148 crystallographic and carbohydrate molecular modeling approaches 86:148 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 4 16 theme crystallographic 503:518 arg1 data 520:523 The crystallographic data 499:523 The crystallographic data 499:523 The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. 29255015 3 17 theme complete 404:411 arg1 glycoprotein 413:424 the complete glycoprotein 400:424 the complete glycoprotein 400:424 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 3 18 theme detailed 376:383 arg1 description 385:395 A detailed description 374:395 A detailed description of the complete glycoprotein 374:424 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 1 19 theme Vp54 192:195 arg1 unusual 286:292 unusual 286:292 unusual 286:292 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 1 19 theme Vp54 192:195 arg1 glycans 155:161 The glycans 151:161 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) 151:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 5 20 theme crystallographic 835:850 arg1 environment 852:862 the crystallographic environment 831:862 the crystallographic environment 831:862 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 6 21 theme molecular 921:929 arg1 modeling 931:938 carbohydrate molecular modeling 908:938 carbohydrate molecular modeling 908:938 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 0 22 theme chlorovirus 17:27 arg1 glycoprotein 49:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 5 23 theme missing 725:731 arg1 monosaccharides 733:747 the missing monosaccharides 721:747 the missing monosaccharides 721:747 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 6 24 gly glycoprotein 1005:1016 arg1 glycoprotein 1005:1016 a glycoprotein 1003:1016 a glycoprotein 1003:1016 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 4 25 theme glycosylation 647:659 arg1 sites 661:665 the glycosylation sites 643:665 the glycosylation sites 643:665 The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. 29255015 2 26 theme reported 343:350 arg1 structure 363:371 the previously reported Vp54 X-ray structure 328:371 the previously reported Vp54 X-ray structure 328:371 This prompted a reexamination of the previously reported Vp54 X-ray structure. 29255015 1 27 theme Paramecium 201:210 arg1 PBCV-1 238:243 PBCV-1 238:243 PBCV-1 238:243 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 1 27 theme Paramecium 201:210 arg1 virus 231:235 Paramecium bursaria chlorella virus 201:235 Paramecium bursaria chlorella virus (PBCV-1) 201:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 0 28 theme major 36:40 arg1 glycoprotein 49:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 4 29 theme protein 585:591 arg1 backbone 593:600 the protein backbone 581:600 the protein backbone 581:600 The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. 29255015 3 30 theme crystallographic 452:467 arg1 data 469:472 crystallographic data 452:472 crystallographic data 452:472 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 1 31 theme bursaria 212:219 arg1 PBCV-1 238:243 PBCV-1 238:243 PBCV-1 238:243 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 1 31 theme bursaria 212:219 arg1 virus 231:235 Paramecium bursaria chlorella virus 201:235 Paramecium bursaria chlorella virus (PBCV-1) 201:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 0 32 theme PBCV-1 29:34 arg1 glycoprotein 49:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 3 33 theme glycoprotein 413:424 arg1 description 385:395 A detailed description 374:395 A detailed description of the complete glycoprotein 374:424 A detailed description of the complete glycoprotein was achieved by combining crystallographic data with molecular modeling. 29255015 6 34 theme carbohydrate 908:919 arg1 modeling 931:938 carbohydrate molecular modeling 908:938 carbohydrate molecular modeling 908:938 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 1 35 theme chlorella 221:229 arg1 PBCV-1 238:243 PBCV-1 238:243 PBCV-1 238:243 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 1 35 theme chlorella 221:229 arg1 virus 231:235 Paramecium bursaria chlorella virus 201:235 Paramecium bursaria chlorella virus (PBCV-1) 201:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 0 36 theme glycoprotein 49:60 arg1 Structure 0:8 Structure 0:8 Structure of the chlorovirus PBCV-1 major capsid glycoprotein 0:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 0 37 theme capsid 42:47 arg1 glycoprotein 49:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 6 38 theme glycoprotein 1005:1016 arg1 structure 990:998 the complete glycosylated structure 964:998 the complete glycosylated structure of a glycoprotein 964:1016 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 4 39 gly glycosylation 647:659 arg2 sites 661:665 the glycosylation sites 643:665 the glycosylation sites 643:665 The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. 29255015 7 40 theme major 1062:1066 arg1 protein 1075:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 40 theme major 1062:1066 arg1 it 1033:1034 it 1033:1034 it 1033:1034 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 4 41 theme located 564:570 arg1 monosaccharides 548:562 the monosaccharides 544:562 the monosaccharides located close to the protein backbone 544:600 The crystallographic data identified most of the monosaccharides located close to the protein backbone, but failed to detect those further from the glycosylation sites. 29255015 6 42 theme X-ray 881:885 arg1 crystallography 887:901 X-ray crystallography 881:901 X-ray crystallography 881:901 Thus, combining X-ray crystallography with carbohydrate molecular modeling resulted in determining the complete glycosylated structure of a glycoprotein. 29255015 7 43 theme capsid 1068:1073 arg1 protein 1075:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 43 theme capsid 1068:1073 arg1 it 1033:1034 it 1033:1034 it 1033:1034 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 1 44 theme virus 231:235 arg1 Vp54 192:195 the major capsid protein (Vp54) 166:196 the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) 166:244 The glycans of the major capsid protein (Vp54) of Paramecium bursaria chlorella virus (PBCV-1) were recently described and found to be unusual. 29255015 7 45 theme chlorovirus 1043:1053 arg1 protein 1075:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 45 theme chlorovirus 1043:1053 arg1 it 1033:1034 it 1033:1034 it 1033:1034 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 46 from protein 1075:1081 arg1 case 1027:1030 this case 1022:1030 this case 1022:1030 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 47 theme PBCV-1 1055:1060 arg1 protein 1075:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 the chlorovirus PBCV-1 major capsid protein 1039:1081 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 7 47 theme PBCV-1 1055:1060 arg1 it 1033:1034 it 1033:1034 it 1033:1034 In this case, it is the chlorovirus PBCV-1 major capsid protein. 29255015 2 48 theme structure 363:371 arg1 reexamination 311:323 a reexamination 309:323 a reexamination of the previously reported Vp54 X-ray structure 309:371 This prompted a reexamination of the previously reported Vp54 X-ray structure. 29255015 5 49 theme whole 799:803 arg1 molecule 805:812 the whole molecule 795:812 the whole molecule 795:812 Molecular modeling complemented this model by adding the missing monosaccharides and examined the conformational preference of the whole molecule, alone or within the crystallographic environment. 29255015 0 50 gly glycoprotein 49:60 arg1 glycoprotein 49:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 the chlorovirus PBCV-1 major capsid glycoprotein 13:60 Structure of the chlorovirus PBCV-1 major capsid glycoprotein determined by combining crystallographic and carbohydrate molecular modeling approaches. 29255015 2 51 theme X-ray 357:361 arg1 structure 363:371 the previously reported Vp54 X-ray structure 328:371 the previously reported Vp54 X-ray structure 328:371 This prompted a reexamination of the previously reported Vp54 X-ray structure. 24393138 2 0 from glycoproteins 445:457 arg1 LC-ESI/TOF-MS 419:431 LC-ESI/TOF-MS 419:431 LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern 419:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 2 1 theme glycan 362:367 arg1 compositions 369:380 glycan compositions 362:380 glycan compositions 362:380 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 1 2 gly glycopeptides 227:239 arg2 glycopeptides 227:239 proteolytic glycopeptides 215:239 proteolytic glycopeptides 215:239 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 1 3 theme biological 147:156 arg1 function 158:165 the biological function 143:165 the biological function of proteins 143:177 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 1 4 theme N- 250:251 arg1 glycans 253:259 (N-)glycans 249:259 (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis 249:339 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 3 5 theme antigen 581:587 arg1 spectra 552:558 isotopically resolved mass spectra 525:558 isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools 525:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 0 6 theme intact 69:74 arg1 glycoprotein 76:87 the intact glycoprotein 65:87 the intact glycoprotein 65:87 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 3 7 theme resolved 538:545 arg1 spectra 552:558 isotopically resolved mass spectra 525:558 isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools 525:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 3 8 theme prostate 563:570 arg1 antigen 581:587 prostate specific antigen 563:587 prostate specific antigen (PSA) using bioinformatics tools 563:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 3 8 theme prostate 563:570 arg1 PSA 590:592 PSA 590:592 PSA 590:592 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 1 9 theme function 158:165 arg1 Glycans 107:113 Glycans 107:113 Glycans 107:113 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 1 9 theme function 158:165 arg1 modulators 129:138 important modulators 119:138 important modulators of the biological function of proteins 119:177 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 3 10 theme specific 572:579 arg1 antigen 581:587 prostate specific antigen 563:587 prostate specific antigen (PSA) using bioinformatics tools 563:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 3 10 theme specific 572:579 arg1 PSA 590:592 PSA 590:592 PSA 590:592 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 1 11 theme proteins 170:177 arg1 function 158:165 the biological function 143:165 the biological function of proteins 143:177 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 0 12 theme Glycan 0:5 arg1 analysis 7:14 Glycan analysis 0:14 Glycan analysis of Prostate Specific Antigen (PSA) 0:49 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 0 13 gly glycoprotein 76:87 arg1 glycoprotein 76:87 the intact glycoprotein 65:87 the intact glycoprotein 65:87 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 0 14 theme Prostate 19:26 arg1 Antigen 37:43 Prostate Specific Antigen 19:43 Prostate Specific Antigen (PSA) 19:49 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 0 14 theme Prostate 19:26 arg1 PSA 46:48 PSA 46:48 PSA 46:48 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 3 15 theme few 637:639 arg1 hours 641:645 a few hours 635:645 a few hours 635:645 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 2 16 theme intact 438:443 arg1 glycoproteins 445:457 intact glycoproteins 438:457 intact glycoproteins even with a very complex glycosylation pattern 438:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 3 17 theme bioinformatics 601:614 arg1 tools 616:620 bioinformatics tools 601:620 bioinformatics tools 601:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 3 18 theme mass 547:550 arg1 spectra 552:558 isotopically resolved mass spectra 525:558 isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools 525:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 0 19 theme Antigen 37:43 arg1 analysis 7:14 Glycan analysis 0:14 Glycan analysis of Prostate Specific Antigen (PSA) 0:49 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 1 20 theme glycosidase 287:297 arg1 treatment 299:307 glycosidase treatment 287:307 glycosidase treatment 287:307 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 3 21 theme glycoforms 677:686 arg1 compositions 658:669 the glycan compositions 647:669 the glycan compositions of 38 glycoforms 647:686 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 2 22 theme glycosylation 484:496 arg1 pattern 498:504 a very complex glycosylation pattern 469:504 a very complex glycosylation pattern 469:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 0 23 theme Specific 28:35 arg1 Antigen 37:43 Prostate Specific Antigen 19:43 Prostate Specific Antigen (PSA) 19:49 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 0 23 theme Specific 28:35 arg1 PSA 46:48 PSA 46:48 PSA 46:48 Glycan analysis of Prostate Specific Antigen (PSA) directly from the intact glycoprotein by HR-ESI/TOF-MS. 24393138 2 24 theme complex 476:482 arg1 pattern 498:504 a very complex glycosylation pattern 469:504 a very complex glycosylation pattern 469:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 2 25 with glycoproteins 445:457 arg1 pattern 498:504 a very complex glycosylation pattern 469:504 a very complex glycosylation pattern 469:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 1 26 theme proteolytic 215:225 arg1 glycopeptides 227:239 proteolytic glycopeptides 215:239 proteolytic glycopeptides 215:239 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 3 27 theme glycan 651:656 arg1 compositions 658:669 the glycan compositions 647:669 the glycan compositions of 38 glycoforms 647:686 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 24393138 1 28 theme important 119:127 arg1 Glycans 107:113 Glycans 107:113 Glycans 107:113 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 1 28 theme important 119:127 arg1 modulators 129:138 important modulators 119:138 important modulators of the biological function of proteins 119:177 Glycans are important modulators of the biological function of proteins and are normally characterized from proteolytic glycopeptides or from (N-)glycans released enzymatically by glycosidase treatment or chemically by hydrazinolysis. 24393138 2 29 gly glycoproteins 445:457 arg1 glycoproteins 445:457 intact glycoproteins 438:457 intact glycoproteins even with a very complex glycosylation pattern 438:504 We demonstrate that glycan compositions can easily be determined directly by LC-ESI/TOF-MS from intact glycoproteins even with a very complex glycosylation pattern. 24393138 3 30 theme spectra 552:558 arg1 Interpretation 507:520 Interpretation 507:520 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools 507:620 Interpretation of isotopically resolved mass spectra of prostate specific antigen (PSA) using bioinformatics tools gives within a few hours the glycan compositions of 38 glycoforms. 25614628 0 0 theme Elizabethkingia 87:101 arg1 meningoseptica 103:116 Elizabethkingia meningoseptica 87:116 Elizabethkingia meningoseptica 87:116 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 5 1 theme PNGase 773:778 arg1 F 780:780 PNGase F 773:780 PNGase F 773:780 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 3 2 theme clinical 495:502 arg1 isolate 504:510 a clinical isolate 493:510 a clinical isolate of Elizabethkingia meningoseptica 493:544 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 1 3 theme oligosaccharide 253:267 arg1 chains 269:274 intact asparagine-linked oligosaccharide chains 228:274 intact asparagine-linked oligosaccharide chains 228:274 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 8 4 theme novel 1151:1155 arg1 N-glycosidase 1169:1181 a novel prokaryotic N-glycosidase 1149:1181 a novel prokaryotic N-glycosidase 1149:1181 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 8 4 theme novel 1151:1155 arg1 protein 1138:1144 the candidate protein 1124:1144 the candidate protein 1124:1144 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 1 5 theme first 157:161 arg1 PNGase 163:168 the first PNGase 153:168 the first PNGase identified in prokaryotic cells 153:200 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 1 5 theme first 157:161 arg1 Peptide 119:125 Peptide 119:125 Peptide:N-glycosidase (PNGase) F 119:150 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 1 6 theme chains 269:274 arg1 removal 217:223 the removal 213:223 the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides 213:314 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 5 7 theme substrate 728:736 arg1 specificity 738:748 substrate specificity 728:748 substrate specificity different from that of PNGase F 728:780 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 6 8 theme crystal 787:793 arg1 structure 795:803 The crystal structure 783:803 The crystal structure of the protein 783:818 The crystal structure of the protein was determined at 1.9 Å resolution. 25614628 6 9 theme protein 812:818 arg1 structure 795:803 The crystal structure 783:803 The crystal structure of the protein 783:818 The crystal structure of the protein was determined at 1.9 Å resolution. 25614628 7 10 theme PNGase 883:888 arg1 F 890:890 PNGase F 883:890 PNGase F 883:890 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 7 11 theme Structural 856:865 arg1 comparison 867:876 Structural comparison 856:876 Structural comparison with PNGase F 856:890 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 5 12 theme different 750:758 arg1 specificity 738:748 substrate specificity 728:748 substrate specificity different from that of PNGase F 728:780 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 7 13 from groove 936:941 arg1 domain 995:1000 an additional bowl-like domain 971:1000 an additional bowl-like domain 971:1000 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 7 13 from groove 936:941 arg1 domain 960:965 the catalytic domain 946:965 the catalytic domain 946:965 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 9 14 theme PNGase 1211:1216 arg1 F-II 1218:1221 PNGase F-II 1211:1221 PNGase F-II 1211:1221 The protein has been named PNGase F-II. 25614628 3 15 theme meningoseptica 531:544 arg1 isolate 504:510 a clinical isolate 493:510 a clinical isolate of Elizabethkingia meningoseptica 493:544 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 3 16 theme novel 412:416 arg1 gene 418:421 a novel gene 410:421 a novel gene encoding a protein with a predicted PNGase domain 410:471 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 7 17 with comparison 867:876 arg1 F 890:890 PNGase F 883:890 PNGase F 883:890 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 8 18 theme structural 1074:1083 arg1 analyses 1100:1107 These structural and functional analyses 1068:1107 These structural and functional analyses 1068:1107 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 1 19 link asparagine-linked 235:251 arg1 chains 269:274 intact asparagine-linked oligosaccharide chains 228:274 intact asparagine-linked oligosaccharide chains 228:274 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 8 20 theme functional 1089:1098 arg1 analyses 1100:1107 These structural and functional analyses 1068:1107 These structural and functional analyses 1068:1107 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 5 21 contain has 724:726 arg1 it 691:692 it 691:692 it 691:692 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 5 21 contain has 724:726 arg2 specificity 738:748 substrate specificity 728:748 substrate specificity different from that of PNGase F 728:780 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 1 22 from glycopeptides 302:314 arg1 removal 217:223 the removal 213:223 the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides 213:314 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 1 23 gly glycoproteins 281:293 arg1 glycoproteins 281:293 glycoproteins 281:293 glycoproteins 281:293 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 8 24 theme prokaryotic 1157:1167 arg1 N-glycosidase 1169:1181 a novel prokaryotic N-glycosidase 1149:1181 a novel prokaryotic N-glycosidase 1149:1181 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 8 24 theme prokaryotic 1157:1167 arg1 protein 1138:1144 the candidate protein 1124:1144 the candidate protein 1124:1144 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 1 25 theme prokaryotic 184:194 arg1 cells 196:200 prokaryotic cells 184:200 prokaryotic cells 184:200 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 5 26 theme PNGase 704:709 arg1 activity 711:718 PNGase activity 704:718 PNGase activity 704:718 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 3 27 theme PNGase 459:464 arg1 domain 466:471 a predicted PNGase domain 447:471 a predicted PNGase domain 447:471 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 7 28 theme unknown 1007:1013 arg1 function 1015:1022 unknown function 1007:1022 unknown function 1007:1022 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 4 29 theme candidate 566:574 arg1 protein 576:582 the candidate protein 562:582 the candidate protein 562:582 In this study, the candidate protein was expressed in vitro and was subjected to biochemical and structural analyses. 25614628 1 30 from glycoproteins 281:293 arg1 removal 217:223 the removal 213:223 the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides 213:314 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 1 31 dep Peptide 119:125 arg1 F 150:150 N-glycosidase (PNGase) F 127:150 Peptide:N-glycosidase (PNGase) F 119:150 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 0 32 theme novel 41:45 arg1 peptide 59:65 a novel prokaryotic peptide 39:65 a novel prokaryotic peptide 39:65 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 7 33 theme protein 1059:1065 arg1 terminus 1033:1040 the N terminus 1027:1040 the N terminus of the candidate protein 1027:1065 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 2 34 theme prokaryotic 380:390 arg1 PNGase 392:397 the sole prokaryotic PNGase 371:397 the sole prokaryotic PNGase 371:397 Since its discovery in 1984, PNGase F has remained as the sole prokaryotic PNGase. 25614628 4 35 theme structural 644:653 arg1 analyses 655:662 biochemical and structural analyses 628:662 biochemical and structural analyses 628:662 In this study, the candidate protein was expressed in vitro and was subjected to biochemical and structural analyses. 25614628 5 36 contain possesses 694:702 arg1 it 691:692 it 691:692 it 691:692 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 5 36 contain possesses 694:702 arg2 activity 711:718 PNGase activity 704:718 PNGase activity 704:718 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 1 37 gly glycopeptides 302:314 arg2 glycopeptides 302:314 glycopeptides 302:314 glycopeptides 302:314 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 3 38 theme predicted 449:457 arg1 domain 466:471 a predicted PNGase domain 447:471 a predicted PNGase domain 447:471 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 0 39 theme peptide 59:65 arg1 characterization 19:34 characterization 19:34 characterization 19:34 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 0 39 theme peptide 59:65 arg1 Identification 0:13 Identification 0:13 Identification 0:13 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 7 40 theme N 1031:1031 arg1 terminus 1033:1040 the N terminus 1027:1040 the N terminus of the candidate protein 1027:1065 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 1 41 theme N-glycosidase 127:139 arg1 F 150:150 N-glycosidase (PNGase) F 127:150 Peptide:N-glycosidase (PNGase) F 119:150 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 0 42 theme prokaryotic 47:57 arg1 peptide 59:65 a novel prokaryotic peptide 39:65 a novel prokaryotic peptide 39:65 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 7 43 theme candidate 1049:1057 arg1 protein 1059:1065 the candidate protein 1045:1065 the candidate protein 1045:1065 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 6 44 theme Å 842:842 arg1 resolution 844:853 1.9 Å resolution 838:853 1.9 Å resolution 838:853 The crystal structure of the protein was determined at 1.9 Å resolution. 25614628 7 45 theme larger 914:919 arg1 groove 936:941 a relatively larger glycan-binding groove 901:941 a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain 901:1000 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 3 46 with protein 434:440 arg1 domain 466:471 a predicted PNGase domain 447:471 a predicted PNGase domain 447:471 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 7 47 theme additional 974:983 arg1 domain 995:1000 an additional bowl-like domain 971:1000 an additional bowl-like domain 971:1000 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 1 48 theme PNGase 142:147 arg1 F 150:150 N-glycosidase (PNGase) F 127:150 Peptide:N-glycosidase (PNGase) F 119:150 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 7 49 theme glycan-binding 921:934 arg1 groove 936:941 a relatively larger glycan-binding groove 901:941 a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain 901:1000 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 8 50 theme candidate 1128:1136 arg1 N-glycosidase 1169:1181 a novel prokaryotic N-glycosidase 1149:1181 a novel prokaryotic N-glycosidase 1149:1181 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 8 50 theme candidate 1128:1136 arg1 protein 1138:1144 the candidate protein 1124:1144 the candidate protein 1124:1144 These structural and functional analyses indicated that the candidate protein is a novel prokaryotic N-glycosidase. 25614628 5 51 from that 765:768 arg1 different 750:758 different 750:758 different 750:758 The results revealed that it possesses PNGase activity and has substrate specificity different from that of PNGase F. 25614628 2 52 theme PNGase 346:351 arg1 F 353:353 PNGase F 346:353 PNGase F 346:353 Since its discovery in 1984, PNGase F has remained as the sole prokaryotic PNGase. 25614628 0 53 from meningoseptica 103:116 arg1 N-glycosidase 68:80 N-glycosidase 68:80 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 0:117 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 4 54 theme biochemical 628:638 arg1 analyses 655:662 biochemical and structural analyses 628:662 biochemical and structural analyses 628:662 In this study, the candidate protein was expressed in vitro and was subjected to biochemical and structural analyses. 25614628 2 55 theme sole 375:378 arg1 PNGase 392:397 the sole prokaryotic PNGase 371:397 the sole prokaryotic PNGase 371:397 Since its discovery in 1984, PNGase F has remained as the sole prokaryotic PNGase. 25614628 1 56 theme intact 228:233 arg1 chains 269:274 intact asparagine-linked oligosaccharide chains 228:274 intact asparagine-linked oligosaccharide chains 228:274 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25614628 7 57 theme catalytic 950:958 arg1 domain 960:965 the catalytic domain 946:965 the catalytic domain 946:965 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 0 58 dep Identification 0:13 arg1 N-glycosidase 68:80 N-glycosidase 68:80 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 0:117 Identification and characterization of a novel prokaryotic peptide: N-glycosidase from Elizabethkingia meningoseptica. 25614628 7 59 theme bowl-like 985:993 arg1 domain 995:1000 an additional bowl-like domain 971:1000 an additional bowl-like domain 971:1000 Structural comparison with PNGase F revealed a relatively larger glycan-binding groove in the catalytic domain and an additional bowl-like domain with unknown function at the N terminus of the candidate protein. 25614628 3 60 theme Elizabethkingia 515:529 arg1 meningoseptica 531:544 Elizabethkingia meningoseptica 515:544 Elizabethkingia meningoseptica 515:544 Recently, a novel gene encoding a protein with a predicted PNGase domain was identified from a clinical isolate of Elizabethkingia meningoseptica. 25614628 2 61 from discovery 327:335 arg1 1984 340:343 1984 340:343 1984 340:343 Since its discovery in 1984, PNGase F has remained as the sole prokaryotic PNGase. 25614628 1 62 theme asparagine-linked 235:251 arg1 chains 269:274 intact asparagine-linked oligosaccharide chains 228:274 intact asparagine-linked oligosaccharide chains 228:274 Peptide:N-glycosidase (PNGase) F, the first PNGase identified in prokaryotic cells, catalyzes the removal of intact asparagine-linked oligosaccharide chains from glycoproteins and/or glycopeptides. 25151378 5 0 theme sulfation 1189:1197 arg1 occurrence 1175:1184 the widespread occurrence 1160:1184 the widespread occurrence of sulfation 1160:1197 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 5 0 theme sulfation 1189:1197 arg1 extension 1146:1154 poly-N-acetyllactosamine extension 1121:1154 poly-N-acetyllactosamine extension 1121:1154 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 6 1 theme protein 1372:1378 arg1 carriers 1380:1387 particular glycan and protein carriers 1350:1387 carriers 1380:1387 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 1 2 theme day 155:157 arg1 analysis 181:188 most current day protein glycosylation analysis 142:188 most current day protein glycosylation analysis 142:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 4 3 theme urgent 617:622 arg1 need 624:627 an urgent need 614:627 an urgent need 614:627 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 4 theme coverage 964:971 arg1 depth 946:950 depth 946:950 depth 946:950 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 4 theme coverage 964:971 arg1 breadth 934:940 the breadth 930:940 the breadth 930:940 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 7 5 theme occupying 1500:1508 arg1 glycans 1510:1516 occupying glycans 1500:1516 occupying glycans 1500:1516 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 1 6 theme protein 159:165 arg1 analysis 181:188 most current day protein glycosylation analysis 142:188 most current day protein glycosylation analysis 142:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 7 7 dep driven 1603:1608 arg1 glycobiology 1590:1601 glycobiology 1590:1601 glycobiology 1590:1601 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 7 8 theme site 1448:1451 arg1 occupancy 1453:1461 site occupancy 1448:1461 site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides 1448:1554 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 3 9 theme all-encompassing 533:548 arg1 glycomics 550:558 all-encompassing glycomics 533:558 all-encompassing glycomics 533:558 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 4 10 dep functions 748:756 arg1 scan 743:746 scan 743:746 scan 743:746 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 7 11 theme direct 1521:1526 arg1 sequencing 1528:1537 direct sequencing 1521:1537 direct sequencing of glycopeptides 1521:1554 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 8 12 theme glycans 1759:1765 arg1 analysis 1733:1740 MS analysis 1730:1740 MS analysis of permethylated glycans occupying the central node 1730:1792 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 7 13 theme glycopeptides 1542:1554 arg1 sequencing 1528:1537 direct sequencing 1521:1537 direct sequencing of glycopeptides 1521:1554 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 6 14 theme particular 1350:1359 arg1 glycan 1361:1366 particular glycan and protein carriers 1350:1387 glycan 1361:1366 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 6 14 theme particular 1350:1359 arg1 latter 1394:1399 latter 1394:1399 latter 1394:1399 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 7 15 theme glycans 1510:1516 arg1 range 1491:1495 the full range 1482:1495 the full range of occupying glycans 1482:1516 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 4 16 theme penetrative 767:777 arg1 acquisition 799:809 more penetrative and purposeful data acquisition 762:809 more penetrative and purposeful data acquisition 762:809 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 17 theme chromatographic 659:673 arg1 separation 675:684 chromatographic separation 659:684 chromatographic separation 659:684 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 1 18 theme Mass 86:89 arg1 technique 124:132 the pivotal technique 112:132 the pivotal technique driving most current day protein glycosylation analysis 112:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 1 18 theme Mass 86:89 arg1 MS 105:106 MS 105:106 MS 105:106 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 1 18 theme Mass 86:89 arg1 spectrometry 91:102 Mass spectrometry 86:102 Mass spectrometry (MS) 86:107 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 7 19 gly glycopeptides 1542:1554 arg2 glycopeptides 1542:1554 glycopeptides 1542:1554 glycopeptides 1542:1554 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 1 20 theme glycosylation 167:179 arg1 analysis 181:188 most current day protein glycosylation analysis 142:188 most current day protein glycosylation analysis 142:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 8 21 theme permethylated 1745:1757 arg1 glycans 1759:1765 permethylated glycans 1745:1765 permethylated glycans occupying the central node 1745:1792 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 5 22 theme widespread 1164:1173 arg1 occurrence 1175:1184 the widespread occurrence 1160:1184 the widespread occurrence of sulfation 1160:1197 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 8 23 from glycomics 1695:1703 arg1 workflows 1680:1688 the required experimental workflows 1654:1688 the required experimental workflows from glycomics to glycoproteomics 1654:1722 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 2 24 theme de 268:269 arg1 identification 276:289 de novo identification 268:289 de novo identification 268:289 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 5 25 dep addressing 1065:1074 arg1 overlooked 1082:1091 overlooked 1082:1091 addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation 1065:1259 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 4 26 theme data 794:797 arg1 acquisition 799:809 more penetrative and purposeful data acquisition 762:809 more penetrative and purposeful data acquisition 762:809 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 27 theme innovative 716:725 arg1 functions 748:756 innovative online LC-MS/MS scan functions 716:756 innovative online LC-MS/MS scan functions 716:756 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 28 theme omic-scale 591:600 arg1 complexity 602:611 the omic-scale complexity 587:611 the omic-scale complexity 587:611 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 7 29 theme driven 1603:1608 arg1 venture 1610:1616 any glycobiology driven venture 1586:1616 any glycobiology driven venture 1586:1616 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 4 30 theme purposeful 783:792 arg1 acquisition 799:809 more penetrative and purposeful data acquisition 762:809 more penetrative and purposeful data acquisition 762:809 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 3 31 theme current 438:444 arg1 advances 446:453 current advances 438:453 current advances 438:453 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 0 32 theme glycosylation 29:41 arg1 analysis 43:50 mass spectrometry-based glycosylation analysis 5:50 mass spectrometry-based glycosylation analysis 5:50 From mass spectrometry-based glycosylation analysis to glycomics and glycoproteomics. 25151378 6 33 theme target 1413:1418 arg1 glycoproteomics 1420:1434 target glycoproteomics 1413:1434 target glycoproteomics 1413:1434 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 4 34 theme automated 884:892 arg1 analysis 899:906 automated data analysis 884:906 automated data analysis 884:906 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 3 35 theme MS-based 474:481 arg1 analysis 497:504 MS-based glycosylation analysis 474:504 MS-based glycosylation analysis 474:504 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 0 36 theme spectrometry-based 10:27 arg1 analysis 43:50 mass spectrometry-based glycosylation analysis 5:50 mass spectrometry-based glycosylation analysis 5:50 From mass spectrometry-based glycosylation analysis to glycomics and glycoproteomics. 25151378 8 37 theme experimental 1667:1678 arg1 workflows 1680:1688 the required experimental workflows 1654:1688 the required experimental workflows from glycomics to glycoproteomics 1654:1722 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 2 38 theme glyco-epitopes 375:388 arg1 mapping 333:339 discovery mapping 323:339 discovery mapping of new or biologically implicated glyco-epitopes 323:388 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 7 39 theme full 1486:1489 arg1 range 1491:1495 the full range 1482:1495 the full range of occupying glycans 1482:1516 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 3 40 theme glycosylation 483:495 arg1 analysis 497:504 MS-based glycosylation analysis 474:504 MS-based glycosylation analysis 474:504 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 8 41 theme required 1658:1665 arg1 workflows 1680:1688 the required experimental workflows 1654:1688 the required experimental workflows from glycomics to glycoproteomics 1654:1722 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 4 42 theme chemical 687:694 arg1 derivatization 696:709 chemical derivatization 687:709 chemical derivatization 687:709 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 5 43 theme variations 1045:1054 arg1 mapping 1025:1031 mapping 1025:1031 mapping 1025:1031 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 5 43 theme variations 1045:1054 arg1 resolution 1010:1019 MS (n) -level resolution 996:1019 MS (n) -level resolution 996:1019 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 4 44 dep tools 848:852 arg1 overcome 857:864 overcome 857:864 to overcome the bottleneck in automated data analysis 854:906 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 3 45 theme limitations 459:469 arg1 account 427:433 an account 424:433 an account of current advances and limitations 424:469 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 3 46 theme advances 446:453 arg1 account 427:433 an account 424:433 an account of current advances and limitations 424:469 This chapter attempts to provide an account of current advances and limitations in MS-based glycosylation analysis as it rapidly evolves into all-encompassing glycomics and glycoproteomics. 25151378 5 47 theme poly-N-acetyllactosamine 1121:1144 arg1 extension 1146:1154 poly-N-acetyllactosamine extension 1121:1154 poly-N-acetyllactosamine extension 1121:1154 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 7 48 theme occupancy 1453:1461 arg1 Mapping 1437:1443 Mapping 1437:1443 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides 1437:1554 Mapping of site occupancy without addressing the full range of occupying glycans by direct sequencing of glycopeptides is essentially inadequate for any glycobiology driven venture. 25151378 5 49 theme malignant 1236:1244 arg1 transformation 1246:1259 malignant transformation 1236:1259 malignant transformation 1236:1259 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 6 50 dep and 1325:1327 arg1 uncovered 1314:1322 uncovered 1314:1322 be uncovered 1311:1322 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 4 51 theme data 894:897 arg1 analysis 899:906 automated data analysis 884:906 automated data analysis 884:906 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 2 52 theme new 344:346 arg1 glyco-epitopes 375:388 new or biologically implicated glyco-epitopes 344:388 new or biologically implicated glyco-epitopes 344:388 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 2 53 theme implicated 364:373 arg1 glyco-epitopes 375:388 new or biologically implicated glyco-epitopes 344:388 new or biologically implicated glyco-epitopes 344:388 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 2 54 theme high 220:223 arg1 sensitivity 225:235 high sensitivity 220:235 high sensitivity 220:235 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 1 55 theme pivotal 116:122 arg1 technique 124:132 the pivotal technique 112:132 the pivotal technique driving most current day protein glycosylation analysis 112:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 1 55 theme pivotal 116:122 arg1 spectrometry 91:102 Mass spectrometry 86:102 Mass spectrometry (MS) 86:107 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 6 56 theme relevance 1301:1309 arg1 glycotopes 1287:1296 functional glycotopes 1276:1296 functional glycotopes of relevance 1276:1309 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 4 57 from bottleneck 870:879 arg1 analysis 899:906 automated data analysis 884:906 automated data analysis 884:906 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 58 theme glycomic 955:962 arg1 coverage 964:971 glycomic coverage 955:971 glycomic coverage 955:971 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 59 theme online 727:732 arg1 functions 748:756 innovative online LC-MS/MS scan functions 716:756 innovative online LC-MS/MS scan functions 716:756 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 60 theme empowering 823:832 arg1 tools 848:852 empowering computational tools 823:852 empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage 823:971 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 5 61 theme structural 1093:1102 arg1 features 1104:1111 structural features 1093:1111 structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation 1093:1197 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 5 61 theme structural 1093:1102 arg1 occurrence 1175:1184 the widespread occurrence 1160:1184 the widespread occurrence of sulfation 1160:1197 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 5 61 theme structural 1093:1102 arg1 extension 1146:1154 poly-N-acetyllactosamine extension 1121:1154 poly-N-acetyllactosamine extension 1121:1154 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 4 62 theme computational 834:846 arg1 tools 848:852 empowering computational tools 823:852 empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage 823:971 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 2 63 theme discovery 323:331 arg1 mapping 333:339 discovery mapping 323:339 discovery mapping of new or biologically implicated glyco-epitopes 323:388 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 25151378 5 64 theme -level 1003:1008 arg1 resolution 1010:1019 MS (n) -level resolution 996:1019 MS (n) -level resolution 996:1019 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 6 65 theme functional 1276:1285 arg1 glycotopes 1287:1296 functional glycotopes 1276:1296 functional glycotopes of relevance 1276:1309 Only then can functional glycotopes of relevance be uncovered, and further localized to particular glycan and protein carriers, the latter by means of target glycoproteomics. 25151378 1 66 theme current 147:153 arg1 analysis 181:188 most current day protein glycosylation analysis 142:188 most current day protein glycosylation analysis 142:188 Mass spectrometry (MS) is the pivotal technique driving most current day protein glycosylation analysis. 25151378 4 67 theme LC-MS/MS 734:741 arg1 functions 748:756 innovative online LC-MS/MS scan functions 716:756 innovative online LC-MS/MS scan functions 716:756 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 8 68 theme central 1781:1787 arg1 node 1789:1792 the central node 1777:1792 the central node 1777:1792 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 8 69 theme MS 1730:1731 arg1 analysis 1733:1740 MS analysis 1730:1740 MS analysis of permethylated glycans occupying the central node 1730:1792 This chapter aims to conceptualize the required experimental workflows from glycomics to glycoproteomics, with MS analysis of permethylated glycans occupying the central node. 25151378 4 70 from advances 647:654 arg1 functions 748:756 innovative online LC-MS/MS scan functions 716:756 innovative online LC-MS/MS scan functions 716:756 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 70 from advances 647:654 arg1 separation 675:684 chromatographic separation 659:684 chromatographic separation 659:684 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 4 70 from advances 647:654 arg1 derivatization 696:709 chemical derivatization 687:709 chemical derivatization 687:709 Given the omic-scale complexity, an urgent need is to orchestrate advances in chromatographic separation, chemical derivatization, and innovative online LC-MS/MS scan functions for more penetrative and purposeful data acquisition, along with empowering computational tools to overcome the bottleneck in automated data analysis, in order to increase the breadth and depth of glycomic coverage. 25151378 5 71 theme isomeric 1036:1043 arg1 variations 1045:1054 isomeric variations 1036:1054 isomeric variations 1036:1054 This entails not only MS (n) -level resolution and mapping of isomeric variations but also addressing often overlooked structural features such as poly-N-acetyllactosamine extension and the widespread occurrence of sulfation in relation to immuno-activation and malignant transformation. 25151378 2 72 dep de 268:269 arg1 novo 271:274 novo 271:274 novo 271:274 It is unrivaled in combining high sensitivity, resolution, and precision for de novo identification and therefore most conducive to discovery mapping of new or biologically implicated glyco-epitopes. 27793989 2 0 theme homophilic 340:349 arg1 interactions 351:362 PECAM-1 homophilic interactions 332:362 PECAM-1 homophilic interactions 332:362 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 1 1 theme major 235:239 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 1 theme major 235:239 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 3 2 theme PECAM-1 562:568 arg1 glycans 570:576 PECAM-1 glycans 562:576 PECAM-1 glycans 562:576 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 2 3 theme Previous 299:306 arg1 studies 308:314 Previous studies 299:314 Previous studies 299:314 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 1 4 theme Endothelial 183:193 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 4 theme Endothelial 183:193 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 4 theme Endothelial 183:193 arg1 PECAM-1 221:227 PECAM-1 221:227 PECAM-1 221:227 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 3 5 from role 862:865 arg1 interactions 921:932 PECAM-1 homophilic interactions 902:932 PECAM-1 homophilic interactions 902:932 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 6 theme asparagine 748:757 arg1 Asn-25 783:788 Asn-25 783:788 Asn-25 783:788 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 6 theme asparagine 748:757 arg1 residue 759:765 the asparagine residue 744:765 the asparagine residue at position 25 (Asn-25) 744:789 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 0 7 theme -mediated 95:103 arg1 Interactions 122:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 1 8 theme Adhesion 200:207 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 8 theme Adhesion 200:207 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 8 theme Adhesion 200:207 arg1 PECAM-1 221:227 PECAM-1 221:227 PECAM-1 221:227 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 0 9 theme Homophilic 111:120 arg1 Interactions 122:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 6 10 theme PECAM-1 1720:1726 arg1 form 1712:1715 this mutant form 1700:1715 this mutant form of PECAM-1 1700:1726 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 6 11 with disruption 1792:1801 arg1 thrombin 1808:1815 thrombin 1808:1815 thrombin 1808:1815 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 3 12 theme molecular 602:610 arg1 mass 612:615 its molecular mass 598:615 its molecular mass 598:615 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 6 13 theme N25Q 1626:1629 arg1 PECAM-1 1631:1637 N25Q PECAM-1 1626:1637 N25Q PECAM-1 1626:1637 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 6 14 theme form 1712:1715 arg1 ability 1689:1695 the ability 1685:1695 the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin 1685:1815 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 5 15 gly glycosylated 1458:1469 arg2 position 1479:1486 this position 1474:1486 this position 1474:1486 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 5 15 gly glycosylated 1458:1469 arg1 form 1430:1433 an N25Q mutant form 1415:1433 an N25Q mutant form of PECAM-1 that is not glycosylated at this position 1415:1486 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 0 16 theme Cell 151:154 arg1 Function 164:171 Endothelial Cell Barrier Function 139:171 Endothelial Cell Barrier Function 139:171 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 3 17 theme solved 632:637 arg1 structure 647:655 the newly solved crystal structure 622:655 the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 622:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 0 18 theme Endothelial 49:59 arg1 Molecule 75:82 Human Platelet Endothelial Cell Adhesion Molecule 1 34:84 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 34:94 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 18 theme Endothelial 49:59 arg1 PECAM-1 87:93 PECAM-1 87:93 PECAM-1 87:93 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 5 19 from integrity 1539:1547 arg1 cells 1578:1582 endothelial cell-like REN cells 1552:1582 endothelial cell-like REN cells 1552:1582 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 4 20 theme bridge 1190:1195 arg1 mutation 1229:1236 mutation 1229:1236 mutation 1229:1236 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 4 20 theme bridge 1190:1195 arg1 formation 1160:1168 the formation 1156:1168 the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding 1156:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 21 theme Adhesion 66:73 arg1 Molecule 75:82 Human Platelet Endothelial Cell Adhesion Molecule 1 34:84 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 34:94 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 21 theme Adhesion 66:73 arg1 PECAM-1 87:93 PECAM-1 87:93 PECAM-1 87:93 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 3 22 theme trans 813:817 arg1 interface 838:846 the trans homophilic-binding interface 809:846 the trans homophilic-binding interface 809:846 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 23 theme molecular 976:984 arg1 studies 994:1000 unbiased molecular docking studies 967:1000 unbiased molecular docking studies 967:1000 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 6 24 theme permeability 1761:1772 arg1 barrier 1774:1780 a permeability barrier 1759:1780 a permeability barrier 1759:1780 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 2 25 theme homology 407:414 arg1 domain 416:421 amino-terminal immunoglobulin homology domain 1 377:423 amino-terminal immunoglobulin homology domain 1 377:423 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 7 26 theme homophilic 2023:2032 arg1 interface 2042:2050 the PECAM-1 homophilic binding interface 2011:2050 the PECAM-1 homophilic binding interface 2011:2050 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 3 27 theme PECAM-1 902:908 arg1 interactions 921:932 PECAM-1 homophilic interactions 902:932 PECAM-1 homophilic interactions 902:932 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 5 28 theme barrier 1384:1390 arg1 function 1392:1399 endothelial barrier function 1372:1399 endothelial barrier function 1372:1399 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 2 29 theme amino-terminal 377:390 arg1 domain 416:421 amino-terminal immunoglobulin homology domain 1 377:423 amino-terminal immunoglobulin homology domain 1 377:423 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 5 30 theme cell-like 1564:1572 arg1 cells 1578:1582 endothelial cell-like REN cells 1552:1582 endothelial cell-like REN cells 1552:1582 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 1 31 theme cell 270:273 arg1 junction 289:296 the endothelial cell intercellular junction 254:296 the endothelial cell intercellular junction 254:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 4 32 theme charged 1027:1033 arg1 moieties 1052:1059 negatively charged α2,3 sialic acid moieties 1016:1059 negatively charged α2,3 sialic acid moieties 1016:1059 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 33 from Function 164:171 arg1 Interactions 122:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 3 34 theme human 660:664 arg1 domain 698:703 human PECAM-1 immunoglobulin homology domain 1 660:705 human PECAM-1 immunoglobulin homology domain 1 660:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 5 35 theme Asn-25 1355:1360 arg1 glycan 1362:1367 the Asn-25 glycan 1351:1367 the Asn-25 glycan 1351:1367 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 0 36 theme Glycans 23:29 arg1 Function 164:171 Endothelial Cell Barrier Function 139:171 Endothelial Cell Barrier Function 139:171 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 36 theme Glycans 23:29 arg1 Role 4:7 The Role 0:7 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 0:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 1 37 theme junction 289:296 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 37 theme junction 289:296 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 7 38 theme cell-cell 1973:1981 arg1 interactions 1983:1994 dynamic endothelial cell-cell interactions 1953:1994 dynamic endothelial cell-cell interactions 1953:1994 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 4 39 theme sialic 1040:1045 arg1 moieties 1052:1059 negatively charged α2,3 sialic acid moieties 1016:1059 negatively charged α2,3 sialic acid moieties 1016:1059 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 5 40 theme vascular 1530:1537 arg1 integrity 1539:1547 vascular integrity 1530:1547 vascular integrity in endothelial cell-like REN cells 1530:1582 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 5 41 theme N25Q 1418:1421 arg1 form 1430:1433 an N25Q mutant form 1415:1433 an N25Q mutant form of PECAM-1 that is not glycosylated at this position 1415:1486 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 2 42 theme PECAM-1 332:338 arg1 interactions 351:362 PECAM-1 homophilic interactions 332:362 PECAM-1 homophilic interactions 332:362 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 0 43 theme Human 34:38 arg1 Molecule 75:82 Human Platelet Endothelial Cell Adhesion Molecule 1 34:84 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 34:94 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 43 theme Human 34:38 arg1 PECAM-1 87:93 PECAM-1 87:93 PECAM-1 87:93 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 7 44 theme dynamic 1953:1959 arg1 interactions 1983:1994 dynamic endothelial cell-cell interactions 1953:1994 dynamic endothelial cell-cell interactions 1953:1994 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 7 45 theme sialic 1890:1895 arg1 glycan 1913:1918 a sialic acid-containing glycan 1888:1918 a sialic acid-containing glycan emanating from Asn-25 1888:1940 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 4 46 theme PECAM-1 1097:1103 arg1 interface 1116:1124 the PECAM-1 homophilic interface 1093:1124 the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding 1093:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 3 47 theme domain 698:703 arg1 structure 647:655 the newly solved crystal structure 622:655 the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 622:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 48 theme homophilic 1301:1310 arg1 binding 1312:1318 PECAM-1-mediated homophilic binding 1284:1318 PECAM-1-mediated homophilic binding 1284:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 2 49 theme permeability 468:479 arg1 barrier 481:487 the vascular permeability barrier 455:487 the vascular permeability barrier 455:487 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 4 50 with bridge 1190:1195 arg1 Lys-89 1221:1226 positively charged Lys-89 1202:1226 positively charged Lys-89 1202:1226 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 4 51 theme α2,3 1035:1038 arg1 moieties 1052:1059 negatively charged α2,3 sialic acid moieties 1016:1059 negatively charged α2,3 sialic acid moieties 1016:1059 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 3 52 located located 794:800 arg1 interface 838:846 the trans homophilic-binding interface 809:846 the trans homophilic-binding interface 809:846 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 52 located located 794:800 arg2 glycan 722:727 a glycan 720:727 a glycan emanating from the asparagine residue at position 25 (Asn-25) 720:789 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 52 located located 794:800 arg2 located 794:800 located 794:800 located 794:800 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 5 53 theme PECAM-1 1438:1444 arg1 form 1430:1433 an N25Q mutant form 1415:1433 an N25Q mutant form of PECAM-1 that is not glycosylated at this position 1415:1486 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 3 54 theme immunoglobulin 674:687 arg1 domain 698:703 human PECAM-1 immunoglobulin homology domain 1 660:705 human PECAM-1 immunoglobulin homology domain 1 660:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 1 55 theme Platelet 174:181 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 55 theme Platelet 174:181 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 55 theme Platelet 174:181 arg1 PECAM-1 221:227 PECAM-1 221:227 PECAM-1 221:227 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 3 56 theme homology 689:696 arg1 domain 698:703 human PECAM-1 immunoglobulin homology domain 1 660:705 human PECAM-1 immunoglobulin homology domain 1 660:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 0 57 theme Trans 105:109 arg1 Interactions 122:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 1 58 theme Cell 195:198 arg1 component 241:249 a major component 233:249 a major component of the endothelial cell intercellular junction 233:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 58 theme Cell 195:198 arg1 Molecule 209:216 Platelet Endothelial Cell Adhesion Molecule 1 174:218 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 174:228 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 1 58 theme Cell 195:198 arg1 PECAM-1 221:227 PECAM-1 221:227 PECAM-1 221:227 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 3 59 theme mass 612:615 arg1 mass 612:615 its molecular mass 598:615 its molecular mass 598:615 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 59 theme mass 612:615 arg1 % 593:593 ∼30% 590:593 ∼30% of its molecular mass 590:615 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 60 theme charged 1213:1219 arg1 Lys-89 1221:1226 positively charged Lys-89 1202:1226 positively charged Lys-89 1202:1226 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 61 theme Endothelial 139:149 arg1 Function 164:171 Endothelial Cell Barrier Function 139:171 Endothelial Cell Barrier Function 139:171 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 4 62 from interface 1116:1124 arg1 orientation 1132:1142 an orientation 1129:1142 an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding 1129:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 63 theme Barrier 156:162 arg1 Function 164:171 Endothelial Cell Barrier Function 139:171 Endothelial Cell Barrier Function 139:171 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 6 64 theme mutant 1705:1710 arg1 form 1712:1715 this mutant form 1700:1715 this mutant form of PECAM-1 1700:1726 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 4 65 theme electrostatic 1176:1188 arg1 bridge 1190:1195 an electrostatic bridge 1173:1195 an electrostatic bridge 1173:1195 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 66 theme Cell 61:64 arg1 Molecule 75:82 Human Platelet Endothelial Cell Adhesion Molecule 1 34:84 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 34:94 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 66 theme Cell 61:64 arg1 PECAM-1 87:93 PECAM-1 87:93 PECAM-1 87:93 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 4 67 theme docking 986:992 arg1 studies 994:1000 unbiased molecular docking studies 967:1000 unbiased molecular docking studies 967:1000 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 4 68 theme possibility 954:964 arg1 support 938:944 support 938:944 support of this possibility 938:964 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 6 69 theme Confocal 1585:1592 arg1 microscopy 1594:1603 Confocal microscopy 1585:1603 Confocal microscopy 1585:1603 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 0 70 from Role 4:7 arg1 Interactions 122:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions 34:133 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 7 71 theme binding 2034:2040 arg1 interface 2042:2050 the PECAM-1 homophilic binding interface 2011:2050 the PECAM-1 homophilic binding interface 2011:2050 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 3 72 theme Asn-25-associated 874:890 arg1 glycan 892:897 an Asn-25-associated glycan 871:897 an Asn-25-associated glycan 871:897 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 2 73 theme thrombotic 543:552 arg1 insult 554:559 inflammatory or thrombotic insult 527:559 inflammatory or thrombotic insult 527:559 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 6 74 theme barrier 1774:1780 arg1 re-establishment 1739:1754 re-establishment 1739:1754 re-establishment of a permeability barrier following disruption with thrombin 1739:1815 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 5 75 theme glycan 1362:1367 arg1 contribution 1335:1346 the contribution 1331:1346 the contribution of the Asn-25 glycan to endothelial barrier function 1331:1399 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 3 76 theme homophilic-binding 819:836 arg1 interface 838:846 the trans homophilic-binding interface 809:846 the trans homophilic-binding interface 809:846 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 7 77 theme PECAM-1 2015:2021 arg1 interface 2042:2050 the PECAM-1 homophilic binding interface 2011:2050 the PECAM-1 homophilic binding interface 2011:2050 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 2 78 theme inflammatory 527:538 arg1 insult 554:559 inflammatory or thrombotic insult 527:559 inflammatory or thrombotic insult 527:559 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 5 79 theme endothelial 1372:1382 arg1 function 1392:1399 endothelial barrier function 1372:1399 endothelial barrier function 1372:1399 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 2 80 theme immunoglobulin 392:405 arg1 domain 416:421 amino-terminal immunoglobulin homology domain 1 377:423 amino-terminal immunoglobulin homology domain 1 377:423 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 6 81 theme cell-cell 1664:1672 arg1 junctions 1674:1682 cell-cell junctions 1664:1682 cell-cell junctions 1664:1682 Confocal microscopy showed that although N25Q PECAM-1 concentrates normally at cell-cell junctions, the ability of this mutant form of PECAM-1 to support re-establishment of a permeability barrier following disruption with thrombin was significantly compromised. 27793989 3 82 theme homophilic 910:919 arg1 interactions 921:932 PECAM-1 homophilic interactions 902:932 PECAM-1 homophilic interactions 902:932 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 83 theme acid 1047:1050 arg1 moieties 1052:1059 negatively charged α2,3 sialic acid moieties 1016:1059 negatively charged α2,3 sialic acid moieties 1016:1059 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 3 84 from position 770:777 arg1 Asn-25 783:788 Asn-25 783:788 Asn-25 783:788 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 3 84 from position 770:777 arg1 residue 759:765 the asparagine residue 744:765 the asparagine residue at position 25 (Asn-25) 744:789 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 0 85 theme Sialylated 12:21 arg1 Glycans 23:29 Sialylated Glycans 12:29 Sialylated Glycans 12:29 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 1 86 theme endothelial 258:268 arg1 junction 289:296 the endothelial cell intercellular junction 254:296 the endothelial cell intercellular junction 254:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 5 87 theme REN 1574:1576 arg1 cells 1578:1582 endothelial cell-like REN cells 1552:1582 endothelial cell-like REN cells 1552:1582 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 1 88 theme intercellular 275:287 arg1 junction 289:296 the endothelial cell intercellular junction 254:296 the endothelial cell intercellular junction 254:296 Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) is a major component of the endothelial cell intercellular junction. 27793989 7 89 theme endothelial 1961:1971 arg1 interactions 1983:1994 dynamic endothelial cell-cell interactions 1953:1994 dynamic endothelial cell-cell interactions 1953:1994 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 2 90 theme barrier 481:487 arg1 maintenance 440:450 maintenance 440:450 maintenance of the vascular permeability barrier 440:487 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 3 91 theme crystal 639:645 arg1 structure 647:655 the newly solved crystal structure 622:655 the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 622:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 92 theme unbiased 967:974 arg1 studies 994:1000 unbiased molecular docking studies 967:1000 unbiased molecular docking studies 967:1000 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 0 93 theme Platelet 40:47 arg1 Molecule 75:82 Human Platelet Endothelial Cell Adhesion Molecule 1 34:84 Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1) 34:94 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 0 93 theme Platelet 40:47 arg1 PECAM-1 87:93 PECAM-1 87:93 PECAM-1 87:93 The Role of Sialylated Glycans in Human Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1)-mediated Trans Homophilic Interactions and Endothelial Cell Barrier Function. 27793989 2 94 theme vascular 459:466 arg1 barrier 481:487 the vascular permeability barrier 455:487 the vascular permeability barrier 455:487 Previous studies have shown that PECAM-1 homophilic interactions, mediated by amino-terminal immunoglobulin homology domain 1, contribute to maintenance of the vascular permeability barrier and to its re-establishment following inflammatory or thrombotic insult. 27793989 4 95 theme PECAM-1-mediated 1284:1299 arg1 binding 1312:1318 PECAM-1-mediated homophilic binding 1284:1318 PECAM-1-mediated homophilic binding 1284:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 27793989 5 96 theme mutant 1423:1428 arg1 form 1430:1433 an N25Q mutant form 1415:1433 an N25Q mutant form of PECAM-1 that is not glycosylated at this position 1415:1486 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 7 97 theme acid-containing 1897:1911 arg1 glycan 1913:1918 a sialic acid-containing glycan 1888:1918 a sialic acid-containing glycan emanating from Asn-25 1888:1940 Taken together, these data suggest that a sialic acid-containing glycan emanating from Asn-25 reinforces dynamic endothelial cell-cell interactions by stabilizing the PECAM-1 homophilic binding interface. 27793989 5 98 theme endothelial 1552:1562 arg1 cells 1578:1582 endothelial cell-like REN cells 1552:1582 endothelial cell-like REN cells 1552:1582 To verify the contribution of the Asn-25 glycan to endothelial barrier function, we generated an N25Q mutant form of PECAM-1 that is not glycosylated at this position and examined its ability to contribute to vascular integrity in endothelial cell-like REN cells. 27793989 3 99 theme PECAM-1 666:672 arg1 domain 698:703 human PECAM-1 immunoglobulin homology domain 1 660:705 human PECAM-1 immunoglobulin homology domain 1 660:705 PECAM-1 glycans account for ∼30% of its molecular mass, and the newly solved crystal structure of human PECAM-1 immunoglobulin homology domain 1 reveals that a glycan emanating from the asparagine residue at position 25 (Asn-25) is located within the trans homophilic-binding interface, suggesting a role for an Asn-25-associated glycan in PECAM-1 homophilic interactions. 27793989 4 100 theme homophilic 1105:1114 arg1 interface 1116:1124 the PECAM-1 homophilic interface 1093:1124 the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding 1093:1318 In support of this possibility, unbiased molecular docking studies revealed that negatively charged α2,3 sialic acid moieties bind tightly to a groove within the PECAM-1 homophilic interface in an orientation that favors the formation of an electrostatic bridge with positively charged Lys-89, mutation of which has been shown previously to disrupt PECAM-1-mediated homophilic binding. 25359621 0 0 theme glycophages 81:91 arg1 arrays 23:28 glycophage arrays 12:28 glycophage arrays 12:28 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 0 0 theme glycophages 81:91 arg1 patterning 67:76 patterning 67:76 patterning 67:76 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 0 0 theme glycophages 81:91 arg1 purification 50:61 purification 50:61 purification 50:61 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 3 1 theme compatible 1089:1098 arg1 glycans 1081:1087 glycans 1081:1087 glycans compatible with microarray patterning 1081:1125 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 2 theme drug 598:601 arg1 discovery 603:611 drug discovery [4] 598:615 drug discovery [4] 598:615 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 3 from glycans 300:306 arg1 systems 342:348 biological systems 331:348 biological systems 331:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 4 dep [6 919:920 arg1 7 923:923 7 923:923 7 923:923 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 5 theme powerful 1008:1015 arg1 method 1021:1026 a potentially powerful new method 994:1026 a potentially powerful new method for facile, scalable production 994:1058 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 6 theme immunity 631:638 arg1 interactions 663:674 host-pathogen interactions 649:674 host-pathogen interactions 649:674 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 6 theme immunity 631:638 arg1 discovery 603:611 drug discovery [4] 598:615 drug discovery [4] 598:615 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 6 theme immunity 631:638 arg1 study 622:626 the study 618:626 the study of immunity [5] 618:642 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 7 theme available 795:803 arg1 methods 787:793 the expensive and complex methods 761:793 the expensive and complex methods available to synthesize glycans 761:825 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 8 theme array 1342:1346 arg1 format 1348:1353 array format 1342:1353 array format 1342:1353 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 2 9 dep sources 911:917 arg1 [6 919:920 [6 919:920 [6 919:920 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 10 dep powerful 470:477 arg1 high-throughput 480:494 high-throughput 480:494 high-throughput 480:494 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 11 theme Biotechnology 944:956 arg1 Journal 958:964 Biotechnology Journal 944:964 Biotechnology Journal 944:964 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 12 theme glycan-protein 516:529 arg1 interactions[1,2 531:546 screening glycan-protein interactions[1,2 506:546 screening glycan-protein interactions[1,2 506:546 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 13 from systems 342:348 arg1 understanding 283:295 our understanding 279:295 our understanding of glycans and their functions in biological systems 279:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 14 theme glycoarray 712:721 arg1 applications 723:734 glycoarray applications 712:734 glycoarray applications 712:734 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 15 theme screening 506:514 arg1 interactions[1,2 531:546 screening glycan-protein interactions[1,2 506:546 screening glycan-protein interactions[1,2 506:546 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 16 theme facile 1032:1037 arg1 production 1049:1058 facile, scalable production 1032:1058 facile, scalable production 1032:1058 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 17 dep coded 256:260 arg1 2 680:680 2 680:680 2 680:680 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 0 18 theme glycophage 12:21 arg1 arrays 23:28 glycophage arrays 12:28 glycophage arrays 12:28 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 0 18 theme glycophage 12:21 arg1 patterning 67:76 patterning 67:76 patterning 67:76 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 0 18 theme glycophage 12:21 arg1 purification 50:61 purification 50:61 purification 50:61 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 3 19 theme new 1017:1019 arg1 method 1021:1026 a potentially powerful new method 994:1026 a potentially powerful new method for facile, scalable production 994:1058 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 4 20 theme [8 1143:1144 arg1 approach 1147:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 2 21 theme functions 318:326 arg1 understanding 283:295 our understanding 279:295 our understanding of glycans and their functions in biological systems 279:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 22 theme glycans 1081:1087 arg1 method 1021:1026 a potentially powerful new method 994:1026 a potentially powerful new method for facile, scalable production 994:1058 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 3 22 theme glycans 1081:1087 arg1 purification 1065:1076 purification 1065:1076 purification of glycans compatible with microarray patterning 1065:1125 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 23 dep discovery 603:611 arg1 [1 676:677 [1 676:677 [1 676:677 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 24 from sources 911:917 arg1 glycans 890:896 identifying and tagging glycans 866:896 identifying and tagging glycans from natural sources [6, 7] 866:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 24 from sources 911:917 arg1 challenges 852:861 the challenges 848:861 the challenges in identifying and tagging glycans from natural sources [6, 7] 848:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 25 theme tagging 882:888 arg1 glycans 890:896 identifying and tagging glycans 866:896 identifying and tagging glycans from natural sources [6, 7] 866:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 26 theme Journal 958:964 arg1 issue 935:939 this issue 930:939 this issue of Biotechnology Journal 930:964 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 27 theme glycan 421:426 arg1 glycoarrays 441:451 glycoarrays 441:451 glycoarrays 441:451 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 27 theme glycan 421:426 arg1 microarrays 428:438 printed glycan microarrays 413:438 printed glycan microarrays (glycoarrays) 413:452 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 28 theme display 1211:1217 arg1 deployment 1179:1188 innovative deployment 1168:1188 innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format 1168:1353 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 2 29 theme glycans 300:306 arg1 understanding 283:295 our understanding 279:295 our understanding of glycans and their functions in biological systems 279:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 30 theme printed 413:419 arg1 glycoarrays 441:451 glycoarrays 441:451 glycoarrays 441:451 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 30 theme printed 413:419 arg1 microarrays 428:438 printed glycan microarrays 413:438 printed glycan microarrays (glycoarrays) 413:452 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 31 theme powerful 470:477 arg1 tools 496:500 powerful, high-throughput tools 470:500 powerful, high-throughput tools for screening glycan-protein interactions[1,2 470:546 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 32 theme identifying 866:876 arg1 glycans 890:896 identifying and tagging glycans 866:896 identifying and tagging glycans from natural sources [6, 7] 866:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 33 from functions 318:326 arg1 systems 342:348 biological systems 331:348 biological systems 331:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 34 theme innovative 1168:1177 arg1 deployment 1179:1188 innovative deployment 1168:1188 innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format 1168:1353 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 3 35 dep Çelik 967:971 arg1 al. 976:978 Çelik et al. 967:978 Çelik et al. [8] 967:982 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 3 35 dep Çelik 967:971 arg1 [8 980:981 [8 980:981 [8 980:981 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 0 36 theme facile 31:36 arg1 production 38:47 facile production 31:47 facile production 31:47 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 4 37 theme displayed 1231:1239 arg1 proteins 1241:1248 the displayed proteins 1227:1248 the displayed proteins 1227:1248 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 4 38 theme Çelik 1128:1132 arg1 approach 1147:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 4 39 theme interest 1289:1296 arg1 glycans 1278:1284 specific glycans 1269:1284 specific glycans of interest (glycophages) 1269:1310 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 1 40 theme widespread 113:122 arg1 modification 143:154 a widespread post-translational modification 111:154 a widespread post-translational modification that plays important roles in health and disease 111:203 Glycosylation is a widespread post-translational modification that plays important roles in health and disease. 25359621 1 40 theme widespread 113:122 arg1 Glycosylation 94:106 Glycosylation 94:106 Glycosylation 94:106 Glycosylation is a widespread post-translational modification that plays important roles in health and disease. 25359621 3 41 dep facile 1032:1037 arg1 scalable 1040:1047 scalable 1040:1047 scalable 1040:1047 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 42 theme natural 903:909 arg1 sources 911:917 natural sources [6, 7] 903:924 natural sources [6, 7] 903:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 43 theme disease 575:581 arg1 detection 583:591 disease detection [3] 575:595 disease detection [3] 575:595 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 44 theme filamentous 1193:1203 arg1 display 1211:1217 filamentous phage display 1193:1217 filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format 1193:1353 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 4 45 theme al.’s 1137:1141 arg1 approach 1147:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 1 46 theme post-translational 124:141 arg1 modification 143:154 a widespread post-translational modification 111:154 a widespread post-translational modification that plays important roles in health and disease 111:203 Glycosylation is a widespread post-translational modification that plays important roles in health and disease. 25359621 1 46 theme post-translational 124:141 arg1 Glycosylation 94:106 Glycosylation 94:106 Glycosylation 94:106 Glycosylation is a widespread post-translational modification that plays important roles in health and disease. 25359621 2 47 theme complex 779:785 arg1 methods 787:793 the expensive and complex methods 761:793 the expensive and complex methods available to synthesize glycans 761:825 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 48 theme specific 1269:1276 arg1 glycans 1278:1284 specific glycans 1269:1284 specific glycans of interest (glycophages) 1269:1310 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 3 49 with compatible 1089:1098 arg1 patterning 1116:1125 microarray patterning 1105:1125 microarray patterning 1105:1125 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 2 50 theme host-pathogen 649:661 arg1 interactions 663:674 host-pathogen interactions 649:674 host-pathogen interactions 649:674 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 51 theme biological 331:340 arg1 systems 342:348 biological systems 331:348 biological systems 331:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 52 from challenges 852:861 arg1 glycans 890:896 identifying and tagging glycans 866:896 identifying and tagging glycans from natural sources [6, 7] 866:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 52 from challenges 852:861 arg1 sources 911:917 natural sources [6, 7] 903:924 natural sources [6, 7] 903:924 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 2 53 theme glycan 209:214 arg1 sequence 216:223 glycan sequence 209:223 glycan sequence 209:223 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 0 54 dep arrays 23:28 arg1 production 38:47 facile production 31:47 facile production 31:47 Introducing glycophage arrays: facile production, purification and patterning of glycophages. 25359621 2 55 theme expensive 765:773 arg1 methods 787:793 the expensive and complex methods 761:793 the expensive and complex methods available to synthesize glycans 761:825 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 3 56 theme microarray 1105:1114 arg1 patterning 1116:1125 microarray patterning 1105:1125 microarray patterning 1105:1125 In this issue of Biotechnology Journal, Çelik et al. [8] introduce a potentially powerful new method for facile, scalable production, and purification of glycans compatible with microarray patterning. 25359621 4 57 theme phage 1205:1209 arg1 display 1211:1217 filamentous phage display 1193:1217 filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format 1193:1353 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 2 58 from understanding 283:295 arg1 systems 342:348 biological systems 331:348 biological systems 331:348 As glycan sequence and structure are not directly coded into the genome, our understanding of glycans and their functions in biological systems is much more primitive than that of DNA and proteins.Recently, printed glycan microarrays (glycoarrays) have emerged as powerful, high-throughput tools for screening glycan-protein interactions[1,2], and have been applied in disease detection [3], drug discovery [4], the study of immunity [5], and host-pathogen interactions [1, 2], among others.Unfortunately, glycoarray applications are currently limited by the expensive and complex methods available to synthesize glycans or alternatively, by the challenges in identifying and tagging glycans from natural sources [6, 7]. 25359621 4 59 theme et 1134:1135 arg1 approach 1147:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach 1128:1154 Çelik et al.’s [8] approach is based on innovative deployment of filamentous phage display so that the displayed proteins can be tagged with specific glycans of interest (glycophages) and subsequently patterned in array format. 25359621 1 60 theme important 167:175 arg1 roles 177:181 important roles 167:181 important roles 167:181 Glycosylation is a widespread post-translational modification that plays important roles in health and disease. 25725350 2 0 theme sialic 310:315 arg1 acid 317:320 sialic acid 310:320 sialic acid 310:320 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 7 1 theme plasma 1078:1083 arg1 B-cells 1085:1091 plasma B-cells 1078:1091 plasma B-cells that synthesize the Fc-glycans 1078:1122 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 3 2 theme well-defined 487:498 arg1 stages 509:514 well-defined clinical stages 487:514 well-defined clinical stages 487:514 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 4 3 theme extra-cellular 606:619 arg1 antigens 621:628 extra-cellular antigens 606:628 extra-cellular antigens 606:628 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 1 4 theme antibodies 152:161 arg1 domain 138:143 the Fc domain 131:143 the Fc domain of IgG antibodies in ALS patients 131:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 4 theme antibodies 152:161 arg1 antibodies 152:161 IgG antibodies 148:161 IgG antibodies in ALS patients 148:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 6 5 theme pre-symptomatic 922:936 arg1 stage 938:942 the pre-symptomatic stage 918:942 the pre-symptomatic stage in the secondary lymphoid organs 918:975 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 4 6 theme Tg 574:575 arg1 mice 577:580 ALS Tg mice 570:580 ALS Tg mice 570:580 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 2 7 contain contains 335:342 arg1 structure 269:277 a bi-antennary structure 254:277 a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2) 254:369 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 2 7 contain contains 335:342 arg2 A2BG2 364:368 A2BG2 364:368 A2BG2 364:368 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 2 7 contain contains 335:342 arg2 GlcNAc 356:361 a bisecting GlcNAc 344:361 a bisecting GlcNAc (A2BG2) 344:369 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 2 8 theme bi-antennary 256:267 arg1 structure 269:277 a bi-antennary structure 254:277 a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2) 254:369 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 1 9 theme ALS 166:168 arg1 patients 170:177 ALS patients 166:177 ALS patients 166:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 8 10 theme motor 1396:1400 arg1 antigens 1409:1416 extracellular motor neuron antigens 1382:1416 extracellular motor neuron antigens 1382:1416 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 1 11 from patients 170:177 arg1 domain 138:143 the Fc domain 131:143 the Fc domain of IgG antibodies in ALS patients 131:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 11 from patients 170:177 arg1 antibodies 152:161 IgG antibodies 148:161 IgG antibodies in ALS patients 148:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 6 12 theme intracellular 892:904 arg1 antigens 906:913 intracellular antigens 892:913 intracellular antigens 892:913 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 10 13 theme glycan 1706:1711 arg1 expression 1713:1722 A2BG2 glycan expression 1700:1722 A2BG2 glycan expression 1700:1722 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 4 14 theme glycan 670:675 arg1 patterns 677:684 different glycan patterns 660:684 different glycan patterns 660:684 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 3 15 dep well-defined 487:498 arg1 clinical 500:507 clinical 500:507 clinical 500:507 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 10 16 from frequency 1674:1682 arg1 domain 1734:1739 the Fc domain 1727:1739 the Fc domain 1727:1739 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 10 17 theme Fc 1731:1732 arg1 domain 1734:1739 the Fc domain 1727:1739 the Fc domain 1727:1739 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 1 18 from domain 138:143 arg1 patients 170:177 ALS patients 166:177 ALS patients 166:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 10 19 from quantity 1688:1695 arg1 domain 1734:1739 the Fc domain 1727:1739 the Fc domain 1727:1739 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 4 20 theme neurons 639:645 arg1 intra- 595:600 intra- 595:600 intra- 595:600 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 4 20 theme neurons 639:645 arg1 antigens 621:628 extra-cellular antigens 606:628 extra-cellular antigens 606:628 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 0 21 theme antibodies 64:73 arg1 antibodies 64:73 IgG antibodies 60:73 IgG antibodies of ALS animals 60:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 0 21 theme antibodies 64:73 arg1 domains 49:55 the Fc domains 42:55 the Fc domains of IgG antibodies of ALS animals 42:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 1 22 theme Fc 135:136 arg1 domain 138:143 the Fc domain 131:143 the Fc domain of IgG antibodies in ALS patients 131:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 22 theme Fc 135:136 arg1 antibodies 152:161 IgG antibodies 148:161 IgG antibodies in ALS patients 148:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 6 23 theme secondary 951:959 arg1 organs 970:975 the secondary lymphoid organs 947:975 the secondary lymphoid organs 947:975 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 0 24 from Development 0:10 arg1 antibodies 64:73 IgG antibodies 60:73 IgG antibodies of ALS animals 60:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 0 24 from Development 0:10 arg1 domains 49:55 the Fc domains 42:55 the Fc domains of IgG antibodies of ALS animals 42:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 10 25 theme poor 1631:1634 arg1 response 1652:1659 the poor systemic immune response 1627:1659 the poor systemic immune response 1627:1659 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 0 26 theme ALS 78:80 arg1 animals 82:88 ALS animals 78:88 ALS animals 78:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 1 27 from glycan 121:126 arg1 domain 138:143 the Fc domain 131:143 the Fc domain of IgG antibodies in ALS patients 131:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 27 from glycan 121:126 arg1 antibodies 152:161 IgG antibodies 148:161 IgG antibodies in ALS patients 148:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 9 28 theme effective 1488:1496 arg1 ADCC 1498:1501 the most effective ADCC 1479:1501 the most effective ADCC of motor neurons 1479:1518 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 1 29 theme unique 114:119 arg1 glycan 121:126 a unique glycan 112:126 a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC) 112:235 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 9 30 located observed 1524:1531 arg1 stage 1544:1548 the end stage 1536:1548 the end stage of the disease 1536:1563 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 9 30 located observed 1524:1531 arg2 ADCC 1498:1501 the most effective ADCC 1479:1501 the most effective ADCC of motor neurons 1479:1518 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 11 31 theme prognostic 1804:1813 arg1 biomarker 1815:1823 a potential prognostic biomarker 1792:1823 a potential prognostic biomarker for ALS 1792:1831 Therefore, A2BG2 is a potential prognostic biomarker for ALS. 25725350 11 31 theme prognostic 1804:1813 arg1 A2BG2 1783:1787 A2BG2 1783:1787 A2BG2 1783:1787 Therefore, A2BG2 is a potential prognostic biomarker for ALS. 25725350 9 32 theme neurons 1512:1518 arg1 ADCC 1498:1501 the most effective ADCC 1479:1501 the most effective ADCC of motor neurons 1479:1518 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 6 33 theme immune 1017:1022 arg1 response 1024:1031 a poor systemic immune response 1001:1031 a poor systemic immune response 1001:1031 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 8 34 theme disease 1322:1328 arg1 progression 1330:1340 disease progression 1322:1340 disease progression 1322:1340 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 0 35 theme stage-dependent 15:29 arg1 glycans 31:37 stage-dependent glycans 15:37 stage-dependent glycans 15:37 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 2 36 contain has 250:252 arg2 structure 269:277 a bi-antennary structure 254:277 a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2) 254:369 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 2 36 contain has 250:252 arg1 glycan 243:248 This glycan 238:248 This glycan 238:248 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 6 37 theme poor 1003:1006 arg1 response 1024:1031 a poor systemic immune response 1001:1031 a poor systemic immune response 1001:1031 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 6 38 from stage 938:942 arg1 organs 970:975 the secondary lymphoid organs 947:975 the secondary lymphoid organs 947:975 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 5 39 theme innate 730:735 arg1 cells 744:748 innate immune cells 730:748 innate immune cells found at the disease onset 730:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 3 40 theme A2BG2 421:425 arg1 expression 427:436 A2BG2 expression 421:436 A2BG2 expression 421:436 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 5 41 theme cells 744:748 arg1 insufficient 781:792 insufficient 781:792 insufficient 781:792 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 5 41 theme cells 744:748 arg1 number 720:725 The increased number 706:725 The increased number of innate immune cells found at the disease onset 706:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 2 42 theme core 294:297 arg1 fucose 299:304 core fucose 294:304 core fucose 294:304 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 5 43 theme disease 763:769 arg1 onset 771:775 the disease onset 759:775 the disease onset 759:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 7 44 theme Tg 1229:1230 arg1 mice 1232:1235 ALS Tg mice 1225:1235 ALS Tg mice 1225:1235 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 4 45 theme IgG 543:545 arg1 antibodies 547:556 the IgG antibodies 539:556 the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease 539:703 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 8 46 theme extracellular 1382:1394 arg1 antigens 1409:1416 extracellular motor neuron antigens 1382:1416 extracellular motor neuron antigens 1382:1416 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 3 47 theme cytotoxicity 446:457 arg1 incidence 408:416 the incidence 404:416 the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages 404:514 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 1 48 theme cell 212:215 arg1 ADCC 231:234 ADCC 231:234 ADCC 231:234 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 48 theme cell 212:215 arg1 cytotoxicity 217:228 antibody-dependent cell cytotoxicity 193:228 antibody-dependent cell cytotoxicity (ADCC) 193:235 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 7 49 theme glycosyltransferase 1169:1187 arg1 pattern 1189:1195 the observed glycosyltransferase pattern 1156:1195 the observed glycosyltransferase pattern 1156:1195 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 7 50 theme B-cells 1085:1091 arg1 glycosyltransferases 1054:1073 the glycosyltransferases 1050:1073 the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans 1050:1122 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 2 51 theme acid 317:320 arg1 residues 322:329 the core fucose and sialic acid residues 290:329 the core fucose and sialic acid residues 290:329 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 6 52 theme IgG 855:857 arg1 antibodies 859:868 IgG antibodies 855:868 IgG antibodies 855:868 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 10 53 dep frequency 1674:1682 arg1 the 1670:1672 the 1670:1672 the 1670:1672 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 3 54 theme ALS 465:467 arg1 conditions 469:478 ALS conditions 465:478 ALS conditions 465:478 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 5 55 theme optimal 807:813 arg1 response 831:838 an optimal systemic T-cell response 804:838 an optimal systemic T-cell response 804:838 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 0 56 theme Fc 46:47 arg1 antibodies 64:73 IgG antibodies 60:73 IgG antibodies of ALS animals 60:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 0 56 theme Fc 46:47 arg1 domains 49:55 the Fc domains 42:55 the Fc domains of IgG antibodies of ALS animals 42:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 10 57 theme A2BG2 1700:1704 arg1 expression 1713:1722 A2BG2 glycan expression 1700:1722 A2BG2 glycan expression 1700:1722 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 1 58 theme IgG 148:150 arg1 antibodies 152:161 IgG antibodies 148:161 IgG antibodies in ALS patients 148:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 8 59 theme neuron 1402:1407 arg1 antigens 1409:1416 extracellular motor neuron antigens 1382:1416 extracellular motor neuron antigens 1382:1416 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 4 60 theme ALS 570:572 arg1 mice 577:580 ALS Tg mice 570:580 ALS Tg mice 570:580 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 2 61 theme fucose 299:304 arg1 residues 322:329 the core fucose and sialic acid residues 290:329 the core fucose and sialic acid residues 290:329 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 5 62 theme systemic 815:822 arg1 response 831:838 an optimal systemic T-cell response 804:838 an optimal systemic T-cell response 804:838 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 8 63 dep ALS 1289:1291 arg1 increased 1307:1315 increased 1307:1315 increased with disease progression 1307:1340 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 10 64 theme expression 1713:1722 arg1 quantity 1688:1695 quantity 1688:1695 quantity 1688:1695 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 10 64 theme expression 1713:1722 arg1 frequency 1674:1682 frequency 1674:1682 frequency 1674:1682 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 8 65 theme final 1440:1444 arg1 stage 1446:1450 the final stage 1436:1450 the final stage of the disease 1436:1465 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 4 66 theme different 660:668 arg1 patterns 677:684 different glycan patterns 660:684 different glycan patterns 660:684 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 0 67 theme IgG 60:62 arg1 antibodies 64:73 IgG antibodies 60:73 IgG antibodies of ALS animals 60:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 1 68 from antibodies 152:161 arg1 patients 170:177 ALS patients 166:177 ALS patients 166:177 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 5 69 theme T-cell 824:829 arg1 response 831:838 an optimal systemic T-cell response 804:838 an optimal systemic T-cell response 804:838 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 8 70 theme A2BG2 1260:1264 arg1 glycan 1266:1271 A2BG2 glycan 1260:1271 A2BG2 glycan 1260:1271 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 7 71 theme observed 1160:1167 arg1 pattern 1189:1195 the observed glycosyltransferase pattern 1156:1195 the observed glycosyltransferase pattern 1156:1195 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 5 72 theme increased 710:718 arg1 insufficient 781:792 insufficient 781:792 insufficient 781:792 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 5 72 theme increased 710:718 arg1 number 720:725 The increased number 706:725 The increased number of innate immune cells found at the disease onset 706:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 6 73 theme lymphoid 961:968 arg1 organs 970:975 the secondary lymphoid organs 947:975 the secondary lymphoid organs 947:975 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 8 74 theme disease 1459:1465 arg1 stage 1446:1450 the final stage 1436:1450 the final stage of the disease 1436:1465 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 4 75 theme motor 633:637 arg1 neurons 639:645 motor neurons 633:645 motor neurons that express different glycan patterns during the disease 633:703 Here, we characterize the IgG antibodies produced in ALS Tg mice by detecting intra- and extra-cellular antigens of motor neurons that express different glycan patterns during the disease. 25725350 0 76 theme animals 82:88 arg1 antibodies 64:73 IgG antibodies 60:73 IgG antibodies of ALS animals 60:88 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 5 77 located found 750:754 arg1 onset 771:775 the disease onset 759:775 the disease onset 759:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 5 77 located found 750:754 arg2 cells 744:748 innate immune cells 730:748 innate immune cells found at the disease onset 730:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 2 78 theme bisecting 346:354 arg1 A2BG2 364:368 A2BG2 364:368 A2BG2 364:368 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 2 78 theme bisecting 346:354 arg1 GlcNAc 356:361 a bisecting GlcNAc 344:361 a bisecting GlcNAc (A2BG2) 344:369 This glycan has a bi-antennary structure that lacks the core fucose and sialic acid residues but contains a bisecting GlcNAc (A2BG2). 25725350 9 79 theme motor 1506:1510 arg1 neurons 1512:1518 motor neurons 1506:1518 motor neurons 1506:1518 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 6 80 theme response 1024:1031 arg1 conditions 987:996 the conditions 983:996 the conditions of a poor systemic immune response 983:1031 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 11 81 theme potential 1794:1802 arg1 biomarker 1815:1823 a potential prognostic biomarker 1792:1823 a potential prognostic biomarker for ALS 1792:1831 Therefore, A2BG2 is a potential prognostic biomarker for ALS. 25725350 11 81 theme potential 1794:1802 arg1 A2BG2 1783:1787 A2BG2 1783:1787 A2BG2 1783:1787 Therefore, A2BG2 is a potential prognostic biomarker for ALS. 25725350 0 82 theme glycans 31:37 arg1 Development 0:10 Development 0:10 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 0:89 Development of stage-dependent glycans on the Fc domains of IgG antibodies of ALS animals. 25725350 6 83 theme systemic 1008:1015 arg1 response 1024:1031 a poor systemic immune response 1001:1031 a poor systemic immune response 1001:1031 Nevertheless, IgG antibodies were produced against intracellular antigens at the pre-symptomatic stage in the secondary lymphoid organs under the conditions of a poor systemic immune response. 25725350 10 84 theme IgG 1591:1593 arg1 antibodies 1595:1604 IgG antibodies 1591:1604 IgG antibodies 1591:1604 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 10 85 theme clinical 1756:1763 arg1 stage 1765:1769 the clinical stage 1752:1769 the clinical stage 1752:1769 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 5 86 theme immune 737:742 arg1 cells 744:748 innate immune cells 730:748 innate immune cells found at the disease onset 730:775 The increased number of innate immune cells found at the disease onset was insufficient to induce an optimal systemic T-cell response. 25725350 9 87 theme end 1540:1542 arg1 stage 1544:1548 the end stage 1536:1548 the end stage of the disease 1536:1563 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 3 88 theme IgG 442:444 arg1 cytotoxicity 446:457 IgG cytotoxicity 442:457 IgG cytotoxicity 442:457 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 3 89 theme expression 427:436 arg1 incidence 408:416 the incidence 404:416 the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages 404:514 Little is known, however, about the incidence of A2BG2 expression and IgG cytotoxicity under ALS conditions within well-defined clinical stages. 25725350 7 90 theme ALS 1225:1227 arg1 mice 1232:1235 ALS Tg mice 1225:1235 ALS Tg mice 1225:1235 Moreover, while the glycosyltransferases of plasma B-cells that synthesize the Fc-glycans were regulated by IL-2 or IL-4, the observed glycosyltransferase pattern did not match that found in ALS Tg mice. 25725350 8 91 theme IgG 1355:1357 arg1 antibodies 1359:1368 the IgG antibodies 1351:1368 the IgG antibodies identifying extracellular motor neuron antigens 1351:1416 We further found that A2BG2 glycan is specific for ALS, its quantity increased with disease progression and that the IgG antibodies identifying extracellular motor neuron antigens were developed at the final stage of the disease. 25725350 1 92 theme antibody-dependent 193:210 arg1 ADCC 231:234 ADCC 231:234 ADCC 231:234 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 1 92 theme antibody-dependent 193:210 arg1 cytotoxicity 217:228 antibody-dependent cell cytotoxicity 193:228 antibody-dependent cell cytotoxicity (ADCC) 193:235 We recently revealed a unique glycan on the Fc domain of IgG antibodies in ALS patients that mediates antibody-dependent cell cytotoxicity (ADCC). 25725350 9 93 theme disease 1557:1563 arg1 stage 1544:1548 the end stage 1536:1548 the end stage of the disease 1536:1563 Therefore, the most effective ADCC of motor neurons was observed at the end stage of the disease. 25725350 10 94 theme systemic 1636:1643 arg1 response 1652:1659 the poor systemic immune response 1627:1659 the poor systemic immune response 1627:1659 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 25725350 10 95 theme immune 1645:1650 arg1 response 1652:1659 the poor systemic immune response 1627:1659 the poor systemic immune response 1627:1659 We conclude that in ALS, IgG antibodies are produced despite the poor systemic immune response and that the frequency and quantity of A2BG2 glycan expression on the Fc domain depends on the clinical stage. 28799853 0 0 theme structural 90:99 arg1 properties 117:126 structural and interfacial properties 90:126 structural and interfacial properties of buckwheat protein isolates 90:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 4 1 theme feature 624:630 arg1 analyses 632:639 Structural feature analyses 613:639 Structural feature analyses 613:639 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 5 2 theme BPI-dextran 908:918 arg1 conjugates 920:929 the BPI-dextran conjugates 904:929 the BPI-dextran conjugates obtained by ultrasound treatment 904:962 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 2 3 theme dextran 386:392 arg1 attachment 372:381 The covalent attachment 359:381 The covalent attachment of dextran to BPI 359:399 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 6 4 from stabilizers 1150:1160 arg1 study 1201:1205 this study 1196:1205 this study 1196:1205 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 6 5 theme glycosylation 1087:1099 arg1 Combination 1047:1057 Combination 1047:1057 Combination of ultrasonic treatment and glycosylation 1047:1099 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 6 5 theme glycosylation 1087:1099 arg1 way 1131:1133 an efficient way 1118:1133 an efficient way to develop new stabilizers and thickening agents for food in this study 1118:1205 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 0 6 theme interfacial 105:115 arg1 properties 117:126 structural and interfacial properties 90:126 structural and interfacial properties of buckwheat protein isolates 90:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 0 7 from Effect 0:5 arg1 properties 117:126 structural and interfacial properties 90:126 structural and interfacial properties of buckwheat protein isolates 90:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 4 8 theme random 726:731 arg1 coil 733:736 less α-helix and more random coil 704:736 less α-helix and more random coil 704:736 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 1 9 theme Maillard 294:301 arg1 conjugates 312:321 the Maillard reaction conjugates 290:321 the Maillard reaction conjugates of buckwheat protein isolate (BPI) 290:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 6 10 theme treatment 1073:1081 arg1 Combination 1047:1057 Combination 1047:1057 Combination of ultrasonic treatment and glycosylation 1047:1099 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 6 10 theme treatment 1073:1081 arg1 way 1131:1133 an efficient way 1118:1133 an efficient way to develop new stabilizers and thickening agents for food in this study 1118:1205 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 3 11 theme classical 594:602 arg1 heating 604:610 classical heating 594:610 classical heating 594:610 Emulsifying properties of the conjugates obtained by ultrasound treatment were improved as compared to those obtained by classical heating. 28799853 1 12 theme reaction 303:310 arg1 conjugates 312:321 the Maillard reaction conjugates 290:321 the Maillard reaction conjugates of buckwheat protein isolate (BPI) 290:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 2 13 attach attachment 372:381 arg2 dextran 386:392 dextran 386:392 dextran 386:392 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 2 13 attach attachment 372:381 arg1 BPI 397:399 BPI 397:399 BPI 397:399 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 1 14 gly glycosylation 228:240 arg1 conjugates 312:321 the Maillard reaction conjugates 290:321 the Maillard reaction conjugates of buckwheat protein isolate (BPI) 290:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 2 15 theme Fourier-transform 437:453 arg1 spectra 464:470 the Fourier-transform infrared spectra 433:470 the Fourier-transform infrared spectra 433:470 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 3 16 theme Emulsifying 473:483 arg1 properties 485:494 Emulsifying properties 473:494 Emulsifying properties of the conjugates obtained by ultrasound treatment 473:545 Emulsifying properties of the conjugates obtained by ultrasound treatment were improved as compared to those obtained by classical heating. 28799853 5 17 theme activity 869:876 arg1 measurement 878:888 The surface activity measurement 857:888 The surface activity measurement 857:888 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 4 18 theme classical 838:846 arg1 heating 848:854 classical heating 838:854 classical heating 838:854 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 19 theme more 721:724 arg1 coil 733:736 less α-helix and more random coil 704:736 less α-helix and more random coil 704:736 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 6 20 from agents 1177:1182 arg1 study 1201:1205 this study 1196:1205 this study 1196:1205 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 4 21 theme Structural 613:622 arg1 analyses 632:639 Structural feature analyses 613:639 Structural feature analyses 613:639 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 6 22 theme thickening 1166:1175 arg1 agents 1177:1182 thickening agents 1166:1182 thickening agents 1166:1182 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 6 23 theme ultrasonic 1062:1071 arg1 treatment 1073:1081 ultrasonic treatment 1062:1081 ultrasonic treatment 1062:1081 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 1 24 theme high-intensity 198:211 arg1 ultrasound 213:222 high-intensity ultrasound 198:222 high-intensity ultrasound 198:222 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 1 25 theme conjugates 312:321 arg1 properties 276:285 the structural and interfacial properties 245:285 the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 245:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 4 26 theme tertiary 786:793 arg1 structure 795:803 less compact tertiary structure 773:803 less compact tertiary structure 773:803 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 5 27 theme ultrasound 943:952 arg1 treatment 954:962 ultrasound treatment 943:962 ultrasound treatment 943:962 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 1 28 theme ultrasound 213:222 arg1 effects 187:193 the effects 183:193 the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 183:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 0 29 theme protein 141:147 arg1 isolates 149:156 buckwheat protein isolates 131:156 buckwheat protein isolates 131:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 5 30 theme small 1018:1022 arg1 area 1024:1027 a small area 1016:1027 a small area of the interface 1016:1044 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 2 31 theme infrared 455:462 arg1 spectra 464:470 the Fourier-transform infrared spectra 433:470 the Fourier-transform infrared spectra 433:470 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 0 32 theme combination 10:20 arg1 Effect 0:5 Effect 0:5 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 0:157 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 1 33 theme buckwheat 326:334 arg1 BPI 353:355 BPI 353:355 BPI 353:355 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 1 33 theme buckwheat 326:334 arg1 isolate 344:350 buckwheat protein isolate 326:350 buckwheat protein isolate (BPI) 326:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 0 34 theme buckwheat 131:139 arg1 isolates 149:156 buckwheat protein isolates 131:156 buckwheat protein isolates 131:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 5 35 theme interface 1036:1044 arg1 area 1024:1027 a small area 1016:1027 a small area of the interface 1016:1044 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 1 36 theme glycosylation 228:240 arg1 effects 187:193 the effects 183:193 the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 183:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 0 37 theme ultrasound 40:49 arg1 treatment 51:59 high-intensity ultrasound treatment 25:59 high-intensity ultrasound treatment 25:59 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 1 38 theme protein 336:342 arg1 BPI 353:355 BPI 353:355 BPI 353:355 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 1 38 theme protein 336:342 arg1 isolate 344:350 buckwheat protein isolate 326:350 buckwheat protein isolate (BPI) 326:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 6 39 theme new 1146:1148 arg1 stabilizers 1150:1160 new stabilizers 1146:1160 new stabilizers 1146:1160 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 0 40 theme high-intensity 25:38 arg1 treatment 51:59 high-intensity ultrasound treatment 25:59 high-intensity ultrasound treatment 25:59 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 1 41 theme isolate 344:350 arg1 conjugates 312:321 the Maillard reaction conjugates 290:321 the Maillard reaction conjugates of buckwheat protein isolate (BPI) 290:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 0 42 theme isolates 149:156 arg1 properties 117:126 structural and interfacial properties 90:126 structural and interfacial properties of buckwheat protein isolates 90:156 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 4 43 contain had 700:702 arg2 structure 795:803 less compact tertiary structure 773:803 less compact tertiary structure 773:803 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 43 contain had 700:702 arg1 conjugates 656:665 conjugates 656:665 conjugates obtained by ultrasound treatment 656:698 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 43 contain had 700:702 arg2 coil 733:736 less α-helix and more random coil 704:736 less α-helix and more random coil 704:736 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 43 contain had 700:702 arg2 hydrophobicity 754:767 higher surface hydrophobicity 739:767 higher surface hydrophobicity 739:767 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 44 theme α-helix 709:715 arg1 coil 733:736 less α-helix and more random coil 704:736 less α-helix and more random coil 704:736 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 5 45 theme surface 861:867 arg1 measurement 878:888 The surface activity measurement 857:888 The surface activity measurement 857:888 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 4 46 theme compact 778:784 arg1 structure 795:803 less compact tertiary structure 773:803 less compact tertiary structure 773:803 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 0 47 theme treatment 51:59 arg1 combination 10:20 combination 10:20 combination of high-intensity ultrasound treatment and dextran glycosylation 10:85 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 2 48 theme covalent 363:370 arg1 attachment 372:381 The covalent attachment 359:381 The covalent attachment of dextran to BPI 359:399 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 1 49 theme structural 249:258 arg1 properties 276:285 the structural and interfacial properties 245:285 the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 245:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 3 50 theme ultrasound 526:535 arg1 treatment 537:545 ultrasound treatment 526:545 ultrasound treatment 526:545 Emulsifying properties of the conjugates obtained by ultrasound treatment were improved as compared to those obtained by classical heating. 28799853 0 51 theme glycosylation 73:85 arg1 combination 10:20 combination 10:20 combination of high-intensity ultrasound treatment and dextran glycosylation 10:85 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 6 52 theme efficient 1121:1129 arg1 Combination 1047:1057 Combination 1047:1057 Combination of ultrasonic treatment and glycosylation 1047:1099 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 6 52 theme efficient 1121:1129 arg1 way 1131:1133 an efficient way 1118:1133 an efficient way to develop new stabilizers and thickening agents for food in this study 1118:1205 Combination of ultrasonic treatment and glycosylation was proved to be an efficient way to develop new stabilizers and thickening agents for food in this study. 28799853 5 53 used occupied 1007:1014 arg2 molecule 998:1005 each molecule 993:1005 each molecule 993:1005 The surface activity measurement revealed that the BPI-dextran conjugates obtained by ultrasound treatment were closely packed and that each molecule occupied a small area of the interface. 28799853 3 54 theme conjugates 503:512 arg1 properties 485:494 Emulsifying properties 473:494 Emulsifying properties of the conjugates obtained by ultrasound treatment 473:545 Emulsifying properties of the conjugates obtained by ultrasound treatment were improved as compared to those obtained by classical heating. 28799853 4 55 theme surface 746:752 arg1 hydrophobicity 754:767 higher surface hydrophobicity 739:767 higher surface hydrophobicity 739:767 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 1 56 theme interfacial 264:274 arg1 properties 276:285 the structural and interfacial properties 245:285 the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 245:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 0 57 theme dextran 65:71 arg1 glycosylation 73:85 dextran glycosylation 65:85 dextran glycosylation 65:85 Effect of combination of high-intensity ultrasound treatment and dextran glycosylation on structural and interfacial properties of buckwheat protein isolates. 28799853 2 58 theme spectra 464:470 arg1 examination 418:428 examination 418:428 examination of the Fourier-transform infrared spectra 418:470 The covalent attachment of dextran to BPI was confirmed by examination of the Fourier-transform infrared spectra. 28799853 1 59 from effects 187:193 arg1 properties 276:285 the structural and interfacial properties 245:285 the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI) 245:356 This study investigated the effects of high-intensity ultrasound and glycosylation on the structural and interfacial properties of the Maillard reaction conjugates of buckwheat protein isolate (BPI). 28799853 4 60 theme higher 739:744 arg1 hydrophobicity 754:767 higher surface hydrophobicity 739:767 higher surface hydrophobicity 739:767 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28799853 4 61 theme ultrasound 679:688 arg1 treatment 690:698 ultrasound treatment 679:698 ultrasound treatment 679:698 Structural feature analyses suggested that conjugates obtained by ultrasound treatment had less α-helix and more random coil, higher surface hydrophobicity and less compact tertiary structure as compared to those obtained by classical heating. 28208257 2 0 theme originator 539:548 arg1 products 555:562 originator drug products 539:562 originator drug products 539:562 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 5 1 theme HILIC 1243:1247 arg1 ability 1232:1238 the ability 1228:1238 the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1228:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 4 2 theme chromatography 921:934 arg1 utility 886:892 the utility 882:892 the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level 882:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 3 theme biosimilars 1089:1099 arg1 B 1182:1182 Erbitux/Cetuximab B 1164:1182 Erbitux/Cetuximab B 1164:1182 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 3 theme biosimilars 1089:1099 arg1 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 3 theme biosimilars 1089:1099 arg1 B 1157:1157 Herceptin/Trastuzumab B 1135:1157 Herceptin/Trastuzumab B 1135:1157 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 3 theme biosimilars 1089:1099 arg1 mAbs 1101:1104 originator and biosimilars mAbs 1074:1104 originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) 1074:1183 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 1 4 theme mAbs 272:275 arg1 approval 239:246 approval 239:246 approval 239:246 The development and approval processes of biosimilar mAbs depend on their comparability to originators. 28208257 1 4 theme mAbs 272:275 arg1 development 223:233 development 223:233 development 223:233 The development and approval processes of biosimilar mAbs depend on their comparability to originators. 28208257 0 5 theme Analytical 85:94 arg1 Tool 96:99 A Powerful Analytical Tool 74:99 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 3 6 gly glycopeptide 820:831 arg2 glycopeptide 820:831 glycopeptide 820:831 glycopeptide 820:831 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 0 7 theme Middle-up 190:198 arg1 Level 200:204 the Middle-up Level 186:204 the Middle-up Level of Analysis 186:216 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 5 8 theme quality 1607:1613 arg1 control 1615:1621 quality control 1607:1621 quality control 1607:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 0 9 dep Originator 123:132 arg1 Antibodies 172:181 Therapeutic Monoclonal Antibodies 149:181 Therapeutic Monoclonal Antibodies 149:181 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 5 10 theme hydrophilic 1260:1270 arg1 variants 1272:1279 hydrophilic variants 1260:1279 hydrophilic variants of protein biopharmaceuticals 1260:1309 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 11 theme powerful 1472:1479 arg1 tool 1492:1495 a powerful analytical tool 1470:1495 a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1470:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 0 12 theme Analysis 209:216 arg1 Level 200:204 the Middle-up Level 186:204 the Middle-up Level of Analysis 186:216 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 5 13 theme analysis 1337:1344 arg1 level 1328:1332 the middle-up level 1314:1332 the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1314:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 4 14 gly glycosylation 1016:1028 arg2 level 1054:1058 the protein level 1042:1058 the protein level 1042:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 15 dep mAbs 1101:1104 arg1 B 1182:1182 Erbitux/Cetuximab B 1164:1182 Erbitux/Cetuximab B 1164:1182 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 15 dep mAbs 1101:1104 arg1 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 15 dep mAbs 1101:1104 arg1 B 1157:1157 Herceptin/Trastuzumab B 1135:1157 Herceptin/Trastuzumab B 1135:1157 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 15 dep mAbs 1101:1104 arg1 mAbs 1101:1104 originator and biosimilars mAbs 1074:1104 originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) 1074:1183 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 16 gly glycosylation 569:581 arg1 mAbs 594:597 mAbs 594:597 mAbs 594:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 5 17 theme hyphenation 1416:1426 arg1 level 1328:1332 the middle-up level 1314:1332 the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1314:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 18 theme originator 1519:1528 arg1 mAbs 1545:1548 originator and biosimilar mAbs 1519:1548 originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1519:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 4 19 theme qualitative 991:1001 arg1 profiling 1003:1011 the qualitative profiling 987:1011 the qualitative profiling of glycosylation patterns at the protein level 987:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 20 theme glycosylation 767:779 arg1 profile 781:787 a glycosylation profile 765:787 a glycosylation profile 765:787 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 4 21 theme protein 1046:1052 arg1 level 1054:1058 the protein level 1042:1058 the protein level 1042:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 22 theme mAbs 594:597 arg1 attribute 649:657 an important critical quality attribute 619:657 an important critical quality attribute (CQA) 619:663 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 22 theme mAbs 594:597 arg1 pattern 583:589 The glycosylation pattern 565:589 The glycosylation pattern of mAbs 565:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 0 23 theme Interaction 12:22 arg1 Chromatography 24:37 Hydrophilic Interaction Chromatography 0:37 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 4 24 theme patterns 1030:1037 arg1 profiling 1003:1011 the qualitative profiling 987:1011 the qualitative profiling of glycosylation patterns at the protein level 987:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 25 theme hydrophilic 897:907 arg1 chromatography 921:934 hydrophilic interaction chromatography 897:934 hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) 897:981 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 25 theme hydrophilic 897:907 arg1 HILIC 937:941 HILIC 937:941 HILIC 937:941 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 26 theme glycosylation 1016:1028 arg1 patterns 1030:1037 glycosylation patterns 1016:1037 glycosylation patterns at the protein level 1016:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 27 theme middle-up 1193:1201 arg1 approach 1203:1210 a middle-up approach 1191:1210 a middle-up approach 1191:1210 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 0 28 theme Hydrophilic 0:10 arg1 Chromatography 24:37 Hydrophilic Interaction Chromatography 0:37 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 4 29 from profiling 1003:1011 arg1 level 1054:1058 the protein level 1042:1058 the protein level 1042:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 5 30 theme mAbs 1545:1548 arg1 comparison 1505:1514 the comparison 1501:1514 the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1501:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 31 theme analytical 1481:1490 arg1 tool 1492:1495 a powerful analytical tool 1470:1495 a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1470:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 2 32 theme analytical 334:343 arg1 comparisons 345:355 analytical comparisons 334:355 analytical comparisons 334:355 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 4 33 theme Erbitux/Cetuximab 1164:1180 arg1 B 1182:1182 Erbitux/Cetuximab B 1164:1182 Erbitux/Cetuximab B 1164:1182 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 33 theme Erbitux/Cetuximab 1164:1180 arg1 mAbs 1101:1104 originator and biosimilars mAbs 1074:1104 originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) 1074:1183 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 34 theme interaction 909:919 arg1 chromatography 921:934 hydrophilic interaction chromatography 897:934 hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) 897:981 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 34 theme interaction 909:919 arg1 HILIC 937:941 HILIC 937:941 HILIC 937:941 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 0 35 theme Hyphenated 39:48 arg1 Chromatography 24:37 Hydrophilic Interaction Chromatography 0:37 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 3 36 theme glycan 809:814 arg1 level 833:837 a glycan and glycopeptide level 807:837 a glycan and glycopeptide level of analysis 807:849 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 2 37 dep assess 373:378 arg1 minimize 455:462 minimize 455:462 minimize the risk of being clinically meaningful differences between biosimilar and originator drug products 455:562 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 5 38 theme phase 1379:1383 arg1 chromatography 1392:1405 reversed phase liquid chromatography 1370:1405 reversed phase liquid chromatography 1370:1405 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 3 39 theme important 622:630 arg1 attribute 649:657 an important critical quality attribute 619:657 an important critical quality attribute (CQA) 619:663 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 39 theme important 622:630 arg1 CQA 660:662 CQA 660:662 CQA 660:662 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 39 theme important 622:630 arg1 pattern 583:589 The glycosylation pattern 565:589 The glycosylation pattern of mAbs 565:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 0 40 from Level 200:204 arg1 Comparison 109:118 the Comparison 105:118 the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis 105:216 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 0 41 theme Originator 123:132 arg1 Comparison 109:118 the Comparison 105:118 the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis 105:216 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 2 42 theme structural 380:389 arg1 features 391:398 structural features 380:398 structural features 380:398 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 4 43 with hyphenated 944:953 arg1 MS 979:980 MS 979:980 MS 979:980 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 43 with hyphenated 944:953 arg1 spectrometry 965:976 mass spectrometry 960:976 mass spectrometry (MS) 960:981 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 44 theme glycosylation 569:581 arg1 attribute 649:657 an important critical quality attribute 619:657 an important critical quality attribute (CQA) 619:663 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 44 theme glycosylation 569:581 arg1 pattern 583:589 The glycosylation pattern 565:589 The glycosylation pattern of mAbs 565:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 0 45 theme Mass 55:58 arg1 Spectrometry 60:71 Mass Spectrometry 55:71 Mass Spectrometry 55:71 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 5 46 theme reversed 1370:1377 arg1 chromatography 1392:1405 reversed phase liquid chromatography 1370:1405 reversed phase liquid chromatography 1370:1405 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 0 47 theme Therapeutic 149:159 arg1 Antibodies 172:181 Therapeutic Monoclonal Antibodies 149:181 Therapeutic Monoclonal Antibodies 149:181 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 0 48 theme Biosimilar 138:147 arg1 Comparison 109:118 the Comparison 105:118 the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis 105:216 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 3 49 theme several 670:676 arg1 approaches 689:698 several analytical approaches 670:698 several analytical approaches 670:698 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 0 50 with Hyphenated 39:48 arg1 Spectrometry 60:71 Mass Spectrometry 55:71 Mass Spectrometry 55:71 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 4 51 from level 1054:1058 arg1 profiling 1003:1011 the qualitative profiling 987:1011 the qualitative profiling of glycosylation patterns at the protein level 987:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 51 from level 1054:1058 arg1 patterns 1030:1037 glycosylation patterns 1016:1037 glycosylation patterns at the protein level 1016:1058 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 52 theme hyphenated 944:953 arg1 chromatography 921:934 hydrophilic interaction chromatography 897:934 hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) 897:981 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 52 theme hyphenated 944:953 arg1 HILIC 937:941 HILIC 937:941 HILIC 937:941 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 53 theme analytical 678:687 arg1 approaches 689:698 several analytical approaches 670:698 several analytical approaches 670:698 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 54 theme critical 632:639 arg1 attribute 649:657 an important critical quality attribute 619:657 an important critical quality attribute (CQA) 619:663 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 54 theme critical 632:639 arg1 CQA 660:662 CQA 660:662 CQA 660:662 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 54 theme critical 632:639 arg1 pattern 583:589 The glycosylation pattern 565:589 The glycosylation pattern of mAbs 565:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 5 55 theme routine 1586:1592 arg1 analyses 1594:1601 routine analyses 1586:1601 routine analyses for quality control 1586:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 2 56 theme meaningful 493:502 arg1 differences 504:514 clinically meaningful differences 482:514 clinically meaningful differences between biosimilar and originator drug products 482:562 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 0 57 theme Monoclonal 161:170 arg1 Antibodies 172:181 Therapeutic Monoclonal Antibodies 149:181 Therapeutic Monoclonal Antibodies 149:181 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 3 58 theme quality 641:647 arg1 attribute 649:657 an important critical quality attribute 619:657 an important critical quality attribute (CQA) 619:663 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 58 theme quality 641:647 arg1 CQA 660:662 CQA 660:662 CQA 660:662 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 3 58 theme quality 641:647 arg1 pattern 583:589 The glycosylation pattern 565:589 The glycosylation pattern of mAbs 565:597 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 4 59 theme Herceptin/Trastuzumab 1135:1155 arg1 B 1157:1157 Herceptin/Trastuzumab B 1135:1157 Herceptin/Trastuzumab B 1135:1157 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 59 theme Herceptin/Trastuzumab 1135:1155 arg1 mAbs 1101:1104 originator and biosimilars mAbs 1074:1104 originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) 1074:1183 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 3 60 theme glycopeptide 820:831 arg1 level 833:837 a glycan and glycopeptide level 807:837 a glycan and glycopeptide level of analysis 807:849 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 4 61 theme mass 960:963 arg1 MS 979:980 MS 979:980 MS 979:980 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 61 theme mass 960:963 arg1 spectrometry 965:976 mass spectrometry 960:976 mass spectrometry (MS) 960:981 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 1 62 dep development 223:233 arg1 The 219:221 The 219:221 The 219:221 The development and approval processes of biosimilar mAbs depend on their comparability to originators. 28208257 1 62 dep development 223:233 arg1 processes 248:256 processes 248:256 processes 248:256 The development and approval processes of biosimilar mAbs depend on their comparability to originators. 28208257 5 63 theme biosimilar 1534:1543 arg1 mAbs 1545:1548 originator and biosimilar mAbs 1519:1548 originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1519:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 64 theme HILIC 1435:1439 arg1 features 1441:1448 HILIC features 1435:1448 HILIC features combined to MS 1435:1463 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 65 theme protein 1284:1290 arg1 biopharmaceuticals 1292:1309 protein biopharmaceuticals 1284:1309 protein biopharmaceuticals 1284:1309 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 0 66 dep Chromatography 24:37 arg1 Tool 96:99 A Powerful Analytical Tool 74:99 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 5 67 theme complementarity 1351:1365 arg1 level 1328:1332 the middle-up level 1314:1332 the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1314:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 68 theme biopharmaceuticals 1292:1309 arg1 variants 1272:1279 hydrophilic variants 1260:1279 hydrophilic variants of protein biopharmaceuticals 1260:1309 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 0 69 theme Powerful 76:83 arg1 Tool 96:99 A Powerful Analytical Tool 74:99 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 0:217 Hydrophilic Interaction Chromatography Hyphenated with Mass Spectrometry: A Powerful Analytical Tool for the Comparison of Originator and Biosimilar Therapeutic Monoclonal Antibodies at the Middle-up Level of Analysis. 28208257 2 70 theme post-translational 404:421 arg1 PTM 438:440 PTM 438:440 PTM 438:440 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 2 70 theme post-translational 404:421 arg1 modifications 423:435 post-translational modifications 404:435 post-translational modifications (PTM) 404:441 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 3 71 theme analysis 842:849 arg1 level 833:837 a glycan and glycopeptide level 807:837 a glycan and glycopeptide level of analysis 807:849 The glycosylation pattern of mAbs is considered to be an important critical quality attribute (CQA), and several analytical approaches have been proposed that facilitate characterizing and monitoring a glycosylation profile, albeit mainly at a glycan and glycopeptide level of analysis. 28208257 2 72 theme drug 550:553 arg1 products 555:562 originator drug products 539:562 originator drug products 539:562 Therefore, analytical comparisons are required to assess structural features and post-translational modifications (PTM) and thereby minimize the risk of being clinically meaningful differences between biosimilar and originator drug products. 28208257 4 73 theme originator 1074:1083 arg1 B 1182:1182 Erbitux/Cetuximab B 1164:1182 Erbitux/Cetuximab B 1164:1182 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 73 theme originator 1074:1083 arg1 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 Remicade/Remsina/Inflectra 1107:1132 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 73 theme originator 1074:1083 arg1 B 1157:1157 Herceptin/Trastuzumab B 1135:1157 Herceptin/Trastuzumab B 1135:1157 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 4 73 theme originator 1074:1083 arg1 mAbs 1101:1104 originator and biosimilars mAbs 1074:1104 originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) 1074:1183 In this study, we demonstrate the utility of hydrophilic interaction chromatography (HILIC) hyphenated with mass spectrometry (MS) for the qualitative profiling of glycosylation patterns at the protein level, by comparing originator and biosimilars mAbs (Remicade/Remsina/Inflectra, Herceptin/Trastuzumab B, and Erbitux/Cetuximab B) using a middle-up approach. 28208257 5 74 theme liquid 1385:1390 arg1 chromatography 1392:1405 reversed phase liquid chromatography 1370:1405 reversed phase liquid chromatography 1370:1405 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 5 75 theme middle-up 1318:1326 arg1 level 1328:1332 the middle-up level 1314:1332 the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control 1314:1621 We demonstrate the ability of HILIC to resolve hydrophilic variants of protein biopharmaceuticals at the middle-up level of analysis, its complementarity to reversed phase liquid chromatography, and its hyphenation to MS. HILIC features combined to MS make a powerful analytical tool for the comparison of originator and biosimilar mAbs that could eventually be applied in routine analyses for quality control. 28208257 1 76 theme biosimilar 261:270 arg1 mAbs 272:275 biosimilar mAbs 261:275 biosimilar mAbs 261:275 The development and approval processes of biosimilar mAbs depend on their comparability to originators. 28895363 1 0 theme mAbs 387:390 arg1 efficacy 375:382 efficacy 375:382 efficacy 375:382 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 1 0 theme mAbs 387:390 arg1 safety 364:369 safety 364:369 safety 364:369 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 8 1 theme cell 1712:1715 arg1 system 1728:1733 CHO cell expression system 1708:1733 CHO cell expression system 1708:1733 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 3 2 theme quadrupole 752:761 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 2 theme quadrupole 752:761 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 6 3 with glycans 1349:1355 arg1 α-Gal 1362:1366 α-Gal 1362:1366 α-Gal 1362:1366 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 7 4 theme neuraminic 1562:1571 arg1 NANA 1579:1582 NANA 1579:1582 NANA 1579:1582 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 4 theme neuraminic 1562:1571 arg1 acid 1573:1576 N-acetyl neuraminic acid 1553:1576 N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1553:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 4 theme neuraminic 1562:1571 arg1 sialylation 1625:1635 the desired, normal human-type sialylation 1594:1635 sialylation 1625:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 5 theme expression 1498:1507 arg1 system 1509:1514 CHO cell expression system 1489:1514 CHO cell expression system containing sialic acid 1489:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 3 6 theme monoclonal 567:576 arg1 antibodies 578:587 anti-EGFR monoclonal antibodies 557:587 anti-EGFR monoclonal antibodies produced by different expression systems 557:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 7 7 theme CHO 1489:1491 arg1 system 1509:1514 CHO cell expression system 1489:1514 CHO cell expression system containing sialic acid 1489:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 8 8 theme active 1798:1803 arg1 reactions 1822:1830 active hypersensitivity reactions 1798:1830 active hypersensitivity reactions 1798:1830 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 8 theme active 1798:1803 arg1 lines 1842:1846 CHO cell lines 1833:1846 CHO cell lines 1833:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 7 9 theme sialic 1527:1532 arg1 acid 1534:1537 sialic acid 1527:1537 sialic acid 1527:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 4 10 theme exoglycosidase 939:952 arg1 digestion 954:962 exoglycosidase digestion 939:962 exoglycosidase digestion method 939:969 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 6 11 theme diverse 1330:1336 arg1 mixture 1338:1344 a much more diverse mixture 1318:1344 a much more diverse mixture of glycans with α-Gal 1318:1366 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 3 12 theme different 601:609 arg1 systems 622:628 different expression systems 601:628 different expression systems 601:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 1 13 theme mAb 197:199 arg1 products 201:208 mAb products 197:208 mAb products 197:208 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 6 14 theme N-glycolylneuraminic 1415:1434 arg1 NGNA 1442:1445 NGNA 1442:1445 NGNA 1442:1445 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 14 theme N-glycolylneuraminic 1415:1434 arg1 acid 1436:1439 N-glycolylneuraminic acid 1415:1439 sialylation N-glycolylneuraminic acid (NGNA) 1403:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 8 15 theme CHO 1833:1835 arg1 reactions 1822:1830 active hypersensitivity reactions 1798:1830 active hypersensitivity reactions 1798:1830 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 15 theme CHO 1833:1835 arg1 lines 1842:1846 CHO cell lines 1833:1846 CHO cell lines 1833:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 6 16 dep sialylation 1403:1413 arg1 NGNA 1442:1445 NGNA 1442:1445 NGNA 1442:1445 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 16 dep sialylation 1403:1413 arg1 acid 1436:1439 N-glycolylneuraminic acid 1415:1439 sialylation N-glycolylneuraminic acid (NGNA) 1403:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 5 17 theme cell 1161:1164 arg1 system 1177:1182 the SP2/0 cell expression system 1151:1182 the SP2/0 cell expression system 1151:1182 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 4 18 from MS/MS 913:917 arg1 combination 922:932 combination 922:932 combination with exoglycosidase digestion method 922:969 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 7 19 contain containing 1516:1525 arg2 acid 1534:1537 sialic acid 1527:1537 sialic acid 1527:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 19 contain containing 1516:1525 arg1 system 1509:1514 CHO cell expression system 1489:1514 CHO cell expression system containing sialic acid 1489:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 6 20 theme undesired 1375:1383 arg1 form 1395:1398 an undesired, aberrant form 1372:1398 an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA) 1372:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 8 21 theme preferred 1859:1867 arg1 potential 1784:1792 very low potential 1775:1792 very low potential for active hypersensitivity reactions, CHO cell lines 1775:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 21 theme preferred 1859:1867 arg1 system 1880:1885 the preferred expression system 1855:1885 the preferred expression system for producing anti-EGFR biobetter 1855:1919 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 1 22 theme expression 137:146 arg1 systems 148:154 mammalian expression systems 127:154 mammalian expression systems 127:154 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 6 23 theme cell 1291:1294 arg1 system 1307:1312 SP2/0 cell expression system 1285:1312 SP2/0 cell expression system 1285:1312 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 0 24 theme different 85:93 arg1 systems 106:112 different expression systems 85:112 different expression systems 85:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 4 25 from energy 855:860 arg1 combination 922:932 combination 922:932 combination with exoglycosidase digestion method 922:969 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 5 26 theme combined 993:1000 arg1 data 1002:1005 The combined data 989:1005 The combined data 989:1005 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 4 27 with combination 922:932 arg1 method 964:969 exoglycosidase digestion method 939:969 exoglycosidase digestion method 939:969 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 0 28 from [Characterization 0:16 arg1 antibody 64:71 an anti-EGFR monoclonal antibody 40:71 an anti-EGFR monoclonal antibody produced by different expression systems 40:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 4 29 theme elevated 836:843 arg1 energy 855:860 An alternating low and elevated collision energy 813:860 An alternating low and elevated collision energy scan, in source collision-induced dissociation 813:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 5 30 theme CHO 1070:1072 arg1 system 1090:1095 CHO cell expression system 1070:1095 CHO cell expression system 1070:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 1 31 theme critical 285:292 arg1 modification 311:322 a critical post-translation modification 283:322 a critical post-translation modification 283:322 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 1 31 theme critical 285:292 arg1 glycosylation 266:278 glycosylation 266:278 glycosylation 266:278 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 1 32 dep safety 364:369 arg1 the 360:362 the 360:362 the 360:362 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 4 33 theme low 828:830 arg1 energy 855:860 An alternating low and elevated collision energy 813:860 An alternating low and elevated collision energy scan, in source collision-induced dissociation 813:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 0 34 theme N-glycosylation 21:35 arg1 [Characterization 0:16 [Characterization 0:16 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems 0:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 5 35 theme anti-EGFR 1039:1047 arg1 antibody 1049:1056 anti-EGFR antibody 1039:1056 anti-EGFR antibody produced by CHO cell expression system 1039:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 1 36 theme systems 148:154 arg1 use 120:122 The use 116:122 The use of mammalian expression systems 116:154 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 2 37 theme expression 461:470 arg1 system 472:477 a different expression system 449:477 a different expression system 449:477 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 3 38 theme free 678:681 arg1 oligosaccharides 683:698 free oligosaccharides 678:698 free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 678:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 0 39 theme monoclonal 53:62 arg1 antibody 64:71 an anti-EGFR monoclonal antibody 40:71 an anti-EGFR monoclonal antibody produced by different expression systems 40:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 5 40 theme glycosylation 1114:1126 arg1 pattern 1103:1109 a pattern 1101:1109 a pattern of glycosylation differing from that of the SP2/0 cell expression system 1101:1182 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 3 41 theme ionization 741:750 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 41 theme ionization 741:750 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 6 42 theme anti-EGFR 1254:1262 arg1 antibody 1264:1271 anti-EGFR antibody 1254:1271 anti-EGFR antibody produced by SP2/0 cell expression system 1254:1312 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 5 43 theme expression 1079:1088 arg1 system 1090:1095 CHO cell expression system 1070:1095 CHO cell expression system 1070:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 8 44 theme anti-EGFR 1677:1685 arg1 antibody 1687:1694 anti-EGFR antibody 1677:1694 anti-EGFR antibody produced by CHO cell expression system 1677:1733 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 3 45 theme chromatography 713:726 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 45 theme chromatography 713:726 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 4 46 theme source 871:876 arg1 dissociation 896:907 source collision-induced dissociation 871:907 source collision-induced dissociation 871:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 8 47 theme better 1744:1749 arg1 tolerance 1760:1768 better clinical tolerance 1744:1768 better clinical tolerance 1744:1768 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 5 48 contain had 1097:1099 arg2 pattern 1103:1109 a pattern 1101:1109 a pattern of glycosylation differing from that of the SP2/0 cell expression system 1101:1182 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 5 48 contain had 1097:1099 arg1 region 1029:1034 the Fab region 1021:1034 the Fab region of anti-EGFR antibody produced by CHO cell expression system 1021:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 1 49 gly heterogeneity 180:192 arg1 products 201:208 mAb products 197:208 mAb products 197:208 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 3 50 theme detailed 507:514 arg1 structures 516:525 The detailed structures 503:525 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems 503:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 7 51 theme N-acetyl 1553:1560 arg1 NANA 1579:1582 NANA 1579:1582 NANA 1579:1582 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 51 theme N-acetyl 1553:1560 arg1 acid 1573:1576 N-acetyl neuraminic acid 1553:1576 N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1553:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 51 theme N-acetyl 1553:1560 arg1 sialylation 1625:1635 the desired, normal human-type sialylation 1594:1635 sialylation 1625:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 4 52 theme digestion 954:962 arg1 method 964:969 exoglycosidase digestion method 939:969 exoglycosidase digestion method 939:969 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 8 53 theme CHO 1708:1710 arg1 system 1728:1733 CHO cell expression system 1708:1733 CHO cell expression system 1708:1733 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 3 54 theme time-of-fight 763:775 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 54 theme time-of-fight 763:775 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 55 theme anti-EGFR 557:565 arg1 antibodies 578:587 anti-EGFR monoclonal antibodies 557:587 anti-EGFR monoclonal antibodies produced by different expression systems 557:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 8 56 theme expression 1717:1726 arg1 system 1728:1733 CHO cell expression system 1708:1733 CHO cell expression system 1708:1733 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 6 57 theme glycans 1349:1355 arg1 form 1395:1398 an undesired, aberrant form 1372:1398 an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA) 1372:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 57 theme glycans 1349:1355 arg1 mixture 1338:1344 a much more diverse mixture 1318:1344 a much more diverse mixture of glycans with α-Gal 1318:1366 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 3 58 theme glycans 541:547 arg1 structures 516:525 The detailed structures 503:525 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems 503:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 8 59 theme low 1780:1782 arg1 potential 1784:1792 very low potential 1775:1792 very low potential for active hypersensitivity reactions, CHO cell lines 1775:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 59 theme low 1780:1782 arg1 system 1880:1885 the preferred expression system 1855:1885 the preferred expression system for producing anti-EGFR biobetter 1855:1919 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 1 60 theme remarkable 169:178 arg1 heterogeneity 180:192 a remarkable heterogeneity 167:192 a remarkable heterogeneity of mAb products 167:208 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 7 61 theme cell 1493:1496 arg1 system 1509:1514 CHO cell expression system 1489:1514 CHO cell expression system containing sialic acid 1489:1537 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 6 62 dep undesired 1375:1383 arg1 aberrant 1386:1393 aberrant 1386:1393 aberrant 1386:1393 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 1 63 contain has 335:337 arg2 impact 350:355 a profound impact 339:355 a profound impact 339:355 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 1 63 contain has 335:337 arg1 it 332:333 it 332:333 it 332:333 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 8 64 theme hypersensitivity 1805:1820 arg1 reactions 1822:1830 active hypersensitivity reactions 1798:1830 active hypersensitivity reactions 1798:1830 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 64 theme hypersensitivity 1805:1820 arg1 lines 1842:1846 CHO cell lines 1833:1846 CHO cell lines 1833:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 1 65 theme products 201:208 arg1 heterogeneity 180:192 a remarkable heterogeneity 167:192 a remarkable heterogeneity of mAb products 167:208 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 6 66 theme sialylation 1403:1413 arg1 form 1395:1398 an undesired, aberrant form 1372:1398 an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA) 1372:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 66 theme sialylation 1403:1413 arg1 mixture 1338:1344 a much more diverse mixture 1318:1344 a much more diverse mixture of glycans with α-Gal 1318:1366 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 5 67 theme SP2/0 1155:1159 arg1 system 1177:1182 the SP2/0 cell expression system 1151:1182 the SP2/0 cell expression system 1151:1182 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 5 68 theme expression 1166:1175 arg1 system 1177:1182 the SP2/0 cell expression system 1151:1182 the SP2/0 cell expression system 1151:1182 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 3 69 theme expression 611:620 arg1 systems 622:628 different expression systems 601:628 different expression systems 601:628 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 8 70 theme cell 1837:1840 arg1 reactions 1822:1830 active hypersensitivity reactions 1798:1830 active hypersensitivity reactions 1798:1830 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 70 theme cell 1837:1840 arg1 lines 1842:1846 CHO cell lines 1833:1846 CHO cell lines 1833:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 1 71 theme mammalian 127:135 arg1 systems 148:154 mammalian expression systems 127:154 mammalian expression systems 127:154 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 7 72 from mAb 1473:1475 arg1 α-Gal 1453:1457 The α-Gal 1449:1457 The α-Gal 1449:1457 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 72 from mAb 1473:1475 arg1 absent 1463:1468 absent 1463:1468 absent 1463:1468 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 6 73 theme expression 1296:1305 arg1 system 1307:1312 SP2/0 cell expression system 1285:1312 SP2/0 cell expression system 1285:1312 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 4 74 dep energy 855:860 arg1 scan 862:865 scan 862:865 scan 862:865 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 2 75 from impact 439:444 arg1 N-glycosylation 486:500 mAb N-glycosylation 482:500 mAb N-glycosylation 482:500 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 6 76 theme SP2/0 1285:1289 arg1 system 1307:1312 SP2/0 cell expression system 1285:1312 SP2/0 cell expression system 1285:1312 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 0 77 theme expression 95:104 arg1 systems 106:112 different expression systems 85:112 different expression systems 85:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 8 78 theme expression 1869:1878 arg1 potential 1784:1792 very low potential 1775:1792 very low potential for active hypersensitivity reactions, CHO cell lines 1775:1846 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 8 78 theme expression 1869:1878 arg1 system 1880:1885 the preferred expression system 1855:1885 the preferred expression system for producing anti-EGFR biobetter 1855:1919 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 2 79 theme mAb 482:484 arg1 N-glycosylation 486:500 mAb N-glycosylation 482:500 mAb N-glycosylation 482:500 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 5 80 theme antibody 1049:1056 arg1 region 1029:1034 the Fab region 1021:1034 the Fab region of anti-EGFR antibody produced by CHO cell expression system 1021:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 7 81 theme human-type 1614:1623 arg1 acid 1573:1576 N-acetyl neuraminic acid 1553:1576 N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1553:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 81 theme human-type 1614:1623 arg1 sialylation 1625:1635 the desired, normal human-type sialylation 1594:1635 sialylation 1625:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 8 82 theme anti-EGFR 1901:1909 arg1 biobetter 1911:1919 anti-EGFR biobetter 1901:1919 anti-EGFR biobetter 1901:1919 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 4 83 theme collision 845:853 arg1 energy 855:860 An alternating low and elevated collision energy 813:860 An alternating low and elevated collision energy scan, in source collision-induced dissociation 813:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 2 84 theme present 397:403 arg1 study 405:409 The present study 393:409 The present study 393:409 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 5 85 theme Fc 1196:1197 arg1 region 1199:1204 the Fc region 1192:1204 the Fc region 1192:1204 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 5 86 theme Fab 1025:1027 arg1 region 1029:1034 the Fab region 1021:1034 the Fab region of anti-EGFR antibody produced by CHO cell expression system 1021:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 6 87 contain had 1314:1316 arg1 antibody 1264:1271 anti-EGFR antibody 1254:1271 anti-EGFR antibody produced by SP2/0 cell expression system 1254:1312 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 87 contain had 1314:1316 arg2 form 1395:1398 an undesired, aberrant form 1372:1398 an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA) 1372:1446 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 6 87 contain had 1314:1316 arg2 mixture 1338:1344 a much more diverse mixture 1318:1344 a much more diverse mixture of glycans with α-Gal 1318:1366 We confirmed that anti-EGFR antibody produced by SP2/0 cell expression system had a much more diverse mixture of glycans with α-Gal and an undesired, aberrant form of sialylation N-glycolylneuraminic acid (NGNA). 28895363 1 88 theme post-translation 294:309 arg1 modification 311:322 a critical post-translation modification 283:322 a critical post-translation modification 283:322 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 1 88 theme post-translation 294:309 arg1 glycosylation 266:278 glycosylation 266:278 glycosylation 266:278 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 3 89 theme oligosaccharides 683:698 arg1 level 669:673 the level 665:673 the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 665:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 4 90 theme alternating 816:826 arg1 energy 855:860 An alternating low and elevated collision energy 813:860 An alternating low and elevated collision energy scan, in source collision-induced dissociation 813:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 0 91 theme anti-EGFR 43:51 arg1 antibody 64:71 an anti-EGFR monoclonal antibody 40:71 an anti-EGFR monoclonal antibody produced by different expression systems 40:112 [Characterization of N-glycosylation in an anti-EGFR monoclonal antibody produced by different expression systems]. 28895363 7 92 theme normal 1607:1612 arg1 acid 1573:1576 N-acetyl neuraminic acid 1553:1576 N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1553:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 92 theme normal 1607:1612 arg1 sialylation 1625:1635 the desired, normal human-type sialylation 1594:1635 sialylation 1625:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 93 from absent 1463:1468 arg1 mAb 1473:1475 mAb 1473:1475 mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1473:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 2 94 theme system 472:477 arg1 impact 439:444 the impact 435:444 the impact of a different expression system on mAb N-glycosylation 435:500 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 4 95 theme collision-induced 878:894 arg1 dissociation 896:907 source collision-induced dissociation 871:907 source collision-induced dissociation 871:907 An alternating low and elevated collision energy scan, in source collision-induced dissociation and MS/MS in combination with exoglycosidase digestion method was also adopted. 28895363 7 96 theme desired 1598:1604 arg1 acid 1573:1576 N-acetyl neuraminic acid 1553:1576 N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation 1553:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 7 96 theme desired 1598:1604 arg1 sialylation 1625:1635 the desired, normal human-type sialylation 1594:1635 sialylation 1625:1635 The α-Gal was absent in mAb produced by CHO cell expression system containing sialic acid predominantly N-acetyl neuraminic acid (NANA) which is the desired, normal human-type sialylation. 28895363 2 97 theme different 451:459 arg1 system 472:477 a different expression system 449:477 a different expression system 449:477 The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. 28895363 3 98 theme electrospray 728:739 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 98 theme electrospray 728:739 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 1 99 theme profound 341:348 arg1 impact 350:355 a profound impact 339:355 a profound impact 339:355 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 5 100 theme cell 1074:1077 arg1 system 1090:1095 CHO cell expression system 1070:1095 CHO cell expression system 1070:1095 The combined data revealed that the Fab region of anti-EGFR antibody produced by CHO cell expression system had a pattern of glycosylation differing from that of the SP2/0 cell expression system whereas the Fc region remained basically unchanged. 28895363 3 101 theme liquid 706:711 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 101 theme liquid 706:711 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 8 102 theme clinical 1751:1758 arg1 tolerance 1760:1768 better clinical tolerance 1744:1768 better clinical tolerance 1744:1768 This study theoretically predicts that anti-EGFR antibody produced by CHO cell expression system may show better clinical tolerance, and very low potential for active hypersensitivity reactions, CHO cell lines can be the preferred expression system for producing anti-EGFR biobetter. 28895363 1 103 theme post-translational 228:245 arg1 modifications 247:259 post-translational modifications 228:259 post-translational modifications 228:259 The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. 28895363 3 104 theme mass 777:780 arg1 spectrometry 782:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry 706:793 liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS) 706:810 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 104 theme mass 777:780 arg1 MS 808:809 LC-ESI-QTof MS 796:809 LC-ESI-QTof MS 796:809 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 28895363 3 105 theme individual 530:539 arg1 glycans 541:547 individual glycans 530:547 individual glycans 530:547 The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS). 24621270 5 0 theme extensive 1177:1185 arg1 procedures 1200:1209 extensive purification procedures 1177:1209 extensive purification procedures 1177:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 6 1 theme A. thaliana 1382:1392 arg1 organs 1394:1399 A. thaliana organs 1382:1399 A. thaliana organs 1382:1399 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 5 2 from sets 1047:1050 arg1 amounts 1082:1088 small amounts 1076:1088 small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1076:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 2 from sets 1047:1050 arg1 materials 1099:1107 plant materials 1093:1107 plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1093:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 2 from sets 1047:1050 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 1 3 theme molecular 260:268 arg1 architectures 270:282 molecular architectures 260:282 molecular architectures 260:282 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 2 4 theme cell 554:557 arg1 walls 559:563 cell walls 554:563 cell walls 554:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 5 5 theme purification 1187:1198 arg1 procedures 1200:1209 extensive purification procedures 1177:1209 extensive purification procedures 1177:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 1 6 theme complex 161:167 arg1 assemblies 191:200 complex, multi-macromolecular assemblies 161:200 complex, multi-macromolecular assemblies of glycans and other molecules 161:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 6 theme complex 161:167 arg1 walls 151:155 Plant cell walls 140:155 Plant cell walls 140:155 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 5 7 theme single 1119:1124 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 2 8 theme mechanical 529:538 arg1 properties 540:549 the biological and mechanical properties 510:549 the biological and mechanical properties of cell walls 510:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 5 9 theme cell-wall 1055:1063 arg1 glycans 1065:1071 cell-wall glycans 1055:1071 cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1055:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 4 10 theme detection 818:826 arg1 tools 828:832 detection tools 818:832 detection tools 818:832 The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. 24621270 4 11 theme glycan-directed 777:791 arg1 antibodies 804:813 glycan-directed monoclonal antibodies 777:813 glycan-directed monoclonal antibodies 777:813 The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. 24621270 5 12 theme thaliana 1147:1154 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 6 13 theme epitope 1235:1241 arg1 chromatography 1253:1266 epitope detection chromatography 1235:1266 epitope detection chromatography 1235:1266 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 5 14 theme motifs 1037:1042 arg1 study 1012:1016 the study 1008:1016 the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1008:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 15 from amounts 1082:1088 arg1 glycans 1065:1071 cell-wall glycans 1055:1071 cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1055:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 15 from amounts 1082:1088 arg1 sets 1047:1050 sets 1047:1050 sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1047:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 16 theme antibodies 990:999 arg1 use 960:962 use 960:962 use of established monoclonal antibodies 960:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 0 17 theme cell-wall 114:122 arg1 glycans 131:137 plant cell-wall matrix glycans 108:137 plant cell-wall matrix glycans 108:137 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 5 18 from glycans 1065:1071 arg1 amounts 1082:1088 small amounts 1076:1088 small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1076:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 18 from glycans 1065:1071 arg1 materials 1099:1107 plant materials 1093:1107 plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1093:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 18 from glycans 1065:1071 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 0 19 gly heterogeneity 69:81 arg1 glycans 131:137 plant cell-wall matrix glycans 108:137 plant cell-wall matrix glycans 108:137 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 5 20 theme Arabidopsis 1135:1145 arg1 thaliana 1147:1154 Arabidopsis thaliana 1135:1154 Arabidopsis thaliana 1135:1154 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 0 21 theme plant 108:112 arg1 glycans 131:137 plant cell-wall matrix glycans 108:137 plant cell-wall matrix glycans 108:137 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 6 22 theme sub-populations 1342:1356 arg1 modulation 1368:1377 their modulation 1362:1377 their modulation in A. thaliana organs 1362:1399 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 6 22 theme sub-populations 1342:1356 arg1 heterogeneity 1282:1294 the heterogeneity 1278:1294 the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations 1278:1356 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 2 23 dep remains 376:382 arg1 relate 489:494 relate 489:494 relate to changes in the biological and mechanical properties of cell walls 489:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 2 24 theme biological 514:523 arg1 properties 540:549 the biological and mechanical properties 510:549 the biological and mechanical properties of cell walls 510:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 0 25 theme detection 8:16 arg1 chromatography 18:31 Epitope detection chromatography 0:31 Epitope detection chromatography 0:31 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 6 26 theme rhamnogalacturonan I 1321:1340 arg1 sub-populations 1342:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 2 27 theme cell-wall 331:339 arg1 glycans 341:347 cell-wall glycans 331:347 cell-wall glycans 331:347 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 2 28 theme glycans 341:347 arg1 chemistry 318:326 the chemistry 314:326 the chemistry of cell-wall glycans 314:347 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 0 29 theme Epitope 0:6 arg1 chromatography 18:31 Epitope detection chromatography 0:31 Epitope detection chromatography 0:31 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 6 30 theme pectic 1314:1319 arg1 sub-populations 1342:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 5 31 theme structural 917:926 arg1 features 928:935 glycan structural features 910:935 glycan structural features (epitopes) provided by use of established monoclonal antibodies 910:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 31 theme structural 917:926 arg1 epitopes 938:945 epitopes 938:945 epitopes 938:945 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 0 32 dep heterogeneity 69:81 arg1 the 54:56 the 54:56 the 54:56 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 6 33 gly heterogeneity 1282:1294 arg1 sub-populations 1342:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 2 34 theme interactions 453:464 arg1 diversity 414:422 the diversity 410:422 the diversity of glycan configurations and interactions in muro 410:472 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 5 35 theme high 872:875 arg1 sensitivity 877:887 high sensitivity 872:887 high sensitivity 872:887 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 0 36 theme glycans 131:137 arg1 heterogeneity 69:81 structural heterogeneity 58:81 structural heterogeneity 58:81 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 0 36 theme glycans 131:137 arg1 inter-connections 87:103 inter-connections 87:103 inter-connections 87:103 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 4 37 theme antibodies 804:813 arg1 use 770:772 use 770:772 use of glycan-directed monoclonal antibodies as detection tools 770:832 The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. 24621270 2 38 theme configurations 434:447 arg1 diversity 414:422 the diversity 410:422 the diversity of glycan configurations and interactions in muro 410:472 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 1 39 dep complex 161:167 arg1 multi-macromolecular 170:189 multi-macromolecular 170:189 multi-macromolecular 170:189 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 6 40 theme detection 1243:1251 arg1 chromatography 1253:1266 epitope detection chromatography 1235:1266 epitope detection chromatography 1235:1266 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 0 41 theme matrix 124:129 arg1 glycans 131:137 plant cell-wall matrix glycans 108:137 plant cell-wall matrix glycans 108:137 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 3 42 theme cell-wall 654:662 arg1 sub-populations 678:692 cell-wall matrix glycan sub-populations 654:692 cell-wall matrix glycan sub-populations 654:692 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 4 43 theme monoclonal 793:802 arg1 antibodies 804:813 glycan-directed monoclonal antibodies 777:813 glycan-directed monoclonal antibodies 777:813 The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. 24621270 2 44 theme glycan 427:432 arg1 configurations 434:447 glycan configurations 427:447 glycan configurations 427:447 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 5 45 theme oligosaccharide 1021:1035 arg1 motifs 1037:1042 oligosaccharide motifs 1021:1042 oligosaccharide motifs 1021:1042 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 3 46 theme matrix 664:669 arg1 sub-populations 678:692 cell-wall matrix glycan sub-populations 654:692 cell-wall matrix glycan sub-populations 654:692 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 5 47 theme small 1076:1080 arg1 amounts 1082:1088 small amounts 1076:1088 small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1076:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 47 theme small 1076:1080 arg1 materials 1099:1107 plant materials 1093:1107 plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1093:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 47 theme small 1076:1080 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 2 48 from diversity 414:422 arg1 muro 469:472 muro 469:472 muro 469:472 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 1 49 theme other 217:221 arg1 molecules 223:231 other molecules 217:231 other molecules 217:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 5 50 theme high 839:842 arg1 capacity 859:866 The high discrimination capacity 835:866 The high discrimination capacity 835:866 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 51 theme established 967:977 arg1 antibodies 990:999 established monoclonal antibodies 967:999 established monoclonal antibodies 967:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 52 theme glycan 910:915 arg1 features 928:935 glycan structural features 910:935 glycan structural features (epitopes) provided by use of established monoclonal antibodies 910:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 52 theme glycan 910:915 arg1 epitopes 938:945 epitopes 938:945 epitopes 938:945 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 1 53 theme molecules 223:231 arg1 assemblies 191:200 complex, multi-macromolecular assemblies 161:200 complex, multi-macromolecular assemblies of glycans and other molecules 161:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 53 theme molecules 223:231 arg1 walls 151:155 Plant cell walls 140:155 Plant cell walls 140:155 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 5 54 theme discrimination 844:857 arg1 capacity 859:866 The high discrimination capacity 835:866 The high discrimination capacity 835:866 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 2 55 from changes 499:505 arg1 properties 540:549 the biological and mechanical properties 510:549 the biological and mechanical properties of cell walls 510:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 0 56 dep method 36:41 arg1 chromatography 18:31 Epitope detection chromatography 0:31 Epitope detection chromatography 0:31 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 2 57 theme walls 559:563 arg1 properties 540:549 the biological and mechanical properties 510:549 the biological and mechanical properties of cell walls 510:563 Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. 24621270 5 58 theme materials 1099:1107 arg1 amounts 1082:1088 small amounts 1076:1088 small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1076:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 58 theme materials 1099:1107 arg1 materials 1099:1107 plant materials 1093:1107 plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1093:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 58 theme materials 1099:1107 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 59 theme plant 1093:1097 arg1 organ 1126:1130 a single organ 1117:1130 a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1117:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 59 theme plant 1093:1097 arg1 materials 1099:1107 plant materials 1093:1107 plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1093:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 4 60 theme chromatographic 737:751 arg1 separations 753:763 chromatographic separations 737:763 chromatographic separations 737:763 The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. 24621270 5 61 from study 1012:1016 arg1 glycans 1065:1071 cell-wall glycans 1055:1071 cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1055:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 61 from study 1012:1016 arg1 sets 1047:1050 sets 1047:1050 sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1047:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 6 62 theme chromatography 1253:1266 arg1 use 1228:1230 the use 1224:1230 the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs 1224:1399 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 3 63 theme inter-connections 698:714 arg1 analysis 642:649 epitope detection chromatography analysis 609:649 epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections 609:714 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 3 64 theme detection 617:625 arg1 analysis 642:649 epitope detection chromatography analysis 609:649 epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections 609:714 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 6 65 theme xyloglucan 1299:1308 arg1 sub-populations 1342:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 xyloglucan and pectic rhamnogalacturonan I sub-populations 1299:1356 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 5 66 theme monoclonal 979:988 arg1 antibodies 990:999 established monoclonal antibodies 967:999 established monoclonal antibodies 967:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 6 67 from heterogeneity 1282:1294 arg1 organs 1394:1399 A. thaliana organs 1382:1399 A. thaliana organs 1382:1399 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 6 68 from modulation 1368:1377 arg1 organs 1394:1399 A. thaliana organs 1382:1399 A. thaliana organs 1382:1399 We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs. 24621270 3 69 theme glycan 671:676 arg1 sub-populations 678:692 cell-wall matrix glycan sub-populations 654:692 cell-wall matrix glycan sub-populations 654:692 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 3 70 theme chromatography 627:640 arg1 analysis 642:649 epitope detection chromatography analysis 609:649 epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections 609:714 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 1 71 theme Plant 140:144 arg1 assemblies 191:200 complex, multi-macromolecular assemblies 161:200 complex, multi-macromolecular assemblies of glycans and other molecules 161:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 71 theme Plant 140:144 arg1 walls 151:155 Plant cell walls 140:155 Plant cell walls 140:155 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 0 72 theme structural 58:67 arg1 heterogeneity 69:81 structural heterogeneity 58:81 structural heterogeneity 58:81 Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans. 24621270 3 73 theme sub-populations 678:692 arg1 analysis 642:649 epitope detection chromatography analysis 609:649 epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections 609:714 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 24621270 5 74 theme glycans 1065:1071 arg1 glycans 1065:1071 cell-wall glycans 1055:1071 cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1055:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 5 74 theme glycans 1065:1071 arg1 sets 1047:1050 sets 1047:1050 sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures 1047:1209 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 1 75 theme glycans 205:211 arg1 assemblies 191:200 complex, multi-macromolecular assemblies 161:200 complex, multi-macromolecular assemblies of glycans and other molecules 161:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 75 theme glycans 205:211 arg1 walls 151:155 Plant cell walls 140:155 Plant cell walls 140:155 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 76 theme cell 146:149 arg1 assemblies 191:200 complex, multi-macromolecular assemblies 161:200 complex, multi-macromolecular assemblies of glycans and other molecules 161:231 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 1 76 theme cell 146:149 arg1 walls 151:155 Plant cell walls 140:155 Plant cell walls 140:155 Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. 24621270 5 77 theme features 928:935 arg1 detection 897:905 the detection 893:905 the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies 893:999 The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. 24621270 3 78 theme epitope 609:615 arg1 analysis 642:649 epitope detection chromatography analysis 609:649 epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections 609:714 Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. 25447136 5 0 theme N-Glycosylation 778:792 arg1 process 794:800 the N-Glycosylation process 774:800 the N-Glycosylation process 774:800 Although, it has been previously shown that N-Glycosylation is essential for motility in S. acidocaldarius, as defects in the N-Glycosylation process resulted in none or reduced motile cells, strains lacking one to all six N-Glycosylation sites within FlaB still remained motile. 25447136 7 1 theme microscopy 1134:1143 arg1 analyses 1145:1152 Transmission electron microscopy analyses 1112:1152 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament 1112:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 2 2 theme proteins 369:376 arg1 aspects 358:364 many biological aspects 342:364 many biological aspects of proteins 342:376 N-Glycosylation has been shown to influence many biological aspects of proteins, like protein folding, stability or activity. 25447136 6 3 from Deletion 932:939 arg1 FlaB 984:987 FlaB 984:987 FlaB 984:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 8 4 theme filament 1359:1366 arg1 assembly 1332:1339 assembly 1332:1339 assembly 1332:1339 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 8 4 theme filament 1359:1366 arg1 stability 1318:1326 stability 1318:1326 stability 1318:1326 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 7 5 theme Transmission 1112:1123 arg1 microscopy 1134:1143 Transmission electron microscopy 1112:1143 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament 1112:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 8 6 theme assembly 1426:1433 arg1 parts 1402:1406 other parts 1396:1406 other parts of the archaellum assembly 1396:1433 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 4 7 theme N-glycan 642:649 arg1 attachment 625:634 the attachment 621:634 the attachment of an N-glycan 621:649 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 7 8 theme electron 1125:1132 arg1 microscopy 1134:1143 Transmission electron microscopy 1112:1143 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament 1112:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 3 9 gly N-glycosylated 527:540 arg1 N-glycosylated 527:540 N-glycosylated 527:540 N-glycosylated 527:540 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 9 gly N-glycosylated 527:540 arg1 FlaB 490:493 the archaellum filament subunit FlaB 458:493 the archaellum filament subunit FlaB of Sulfolobus acidocaldarius 458:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 10 theme filament 473:480 arg1 N-glycosylated 527:540 N-glycosylated 527:540 N-glycosylated 527:540 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 10 theme filament 473:480 arg1 FlaB 490:493 the archaellum filament subunit FlaB 458:493 the archaellum filament subunit FlaB of Sulfolobus acidocaldarius 458:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 7 11 theme structural 1223:1232 arg1 change 1234:1239 no structural change 1220:1239 no structural change in length 1220:1249 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 7 12 theme glycosylated 1161:1172 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 5 13 theme reduced 822:828 arg1 cells 837:841 reduced motile cells 822:841 reduced motile cells 822:841 Although, it has been previously shown that N-Glycosylation is essential for motility in S. acidocaldarius, as defects in the N-Glycosylation process resulted in none or reduced motile cells, strains lacking one to all six N-Glycosylation sites within FlaB still remained motile. 25447136 6 14 theme sites 975:979 arg1 Deletion 932:939 Deletion 932:939 Deletion of the first five N-Glycosylation sites in FlaB 932:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 4 15 mod modified 607:614 arg1 Each 543:546 Each 543:546 Each 543:546 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 4 15 mod modified 607:614 arg1 sites 585:589 the six predicted N-Glycosylation sites 551:589 the six predicted N-Glycosylation sites within FlaB 551:601 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 4 15 mod modified 607:614 arg3 attachment 625:634 the attachment 621:634 the attachment of an N-glycan 621:649 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 5 16 theme N-Glycosylation 875:889 arg1 sites 891:895 all six N-Glycosylation sites 867:895 all six N-Glycosylation sites within FlaB 867:907 Although, it has been previously shown that N-Glycosylation is essential for motility in S. acidocaldarius, as defects in the N-Glycosylation process resulted in none or reduced motile cells, strains lacking one to all six N-Glycosylation sites within FlaB still remained motile. 25447136 5 17 theme motile 830:835 arg1 cells 837:841 reduced motile cells 822:841 reduced motile cells 822:841 Although, it has been previously shown that N-Glycosylation is essential for motility in S. acidocaldarius, as defects in the N-Glycosylation process resulted in none or reduced motile cells, strains lacking one to all six N-Glycosylation sites within FlaB still remained motile. 25447136 4 18 theme N-Glycosylation 569:583 arg1 sites 585:589 the six predicted N-Glycosylation sites 551:589 the six predicted N-Glycosylation sites within FlaB 551:601 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 2 19 theme protein 384:390 arg1 folding 392:398 protein folding 384:398 protein folding 384:398 N-Glycosylation has been shown to influence many biological aspects of proteins, like protein folding, stability or activity. 25447136 0 20 theme archaellum 23:32 arg1 filament 34:41 the archaellum filament 19:41 the archaellum filament 19:41 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 1 21 theme life 292:295 arg1 life 292:295 life 292:295 life 292:295 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 1 21 theme life 292:295 arg1 domains 281:287 all three domains 271:287 all three domains of life 271:295 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 3 22 theme archaellum 462:471 arg1 N-glycosylated 527:540 N-glycosylated 527:540 N-glycosylated 527:540 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 22 theme archaellum 462:471 arg1 FlaB 490:493 the archaellum filament subunit FlaB 458:493 the archaellum filament subunit FlaB of Sulfolobus acidocaldarius 458:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 7 23 theme archaellum 1191:1200 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 6 24 from sites 975:979 arg1 FlaB 984:987 FlaB 984:987 FlaB 984:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 8 25 theme other 1396:1400 arg1 parts 1402:1406 other parts 1396:1406 other parts of the archaellum assembly 1396:1433 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 1 26 theme predominant 208:218 arg1 modifications 238:250 the predominant posttranslational modifications 204:250 the predominant posttranslational modifications 204:250 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 3 27 theme acidocaldarius 509:522 arg1 N-glycosylated 527:540 N-glycosylated 527:540 N-glycosylated 527:540 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 27 theme acidocaldarius 509:522 arg1 FlaB 490:493 the archaellum filament subunit FlaB 458:493 the archaellum filament subunit FlaB of Sulfolobus acidocaldarius 458:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 28 theme Sulfolobus 498:507 arg1 acidocaldarius 509:522 Sulfolobus acidocaldarius 498:522 Sulfolobus acidocaldarius 498:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 5 29 from defects 763:769 arg1 process 794:800 the N-Glycosylation process 774:800 the N-Glycosylation process 774:800 Although, it has been previously shown that N-Glycosylation is essential for motility in S. acidocaldarius, as defects in the N-Glycosylation process resulted in none or reduced motile cells, strains lacking one to all six N-Glycosylation sites within FlaB still remained motile. 25447136 0 30 gly N-Glycosylation 0:14 arg1 filament 34:41 the archaellum filament 19:41 the archaellum filament 19:41 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 6 31 theme sites 1075:1079 arg1 removal 1040:1046 removal 1040:1046 removal of all six N-Glycosylation sites 1040:1079 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 1 32 theme posttranslational 220:236 arg1 modifications 238:250 the predominant posttranslational modifications 204:250 the predominant posttranslational modifications 204:250 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 0 33 theme filament 34:41 arg1 N-Glycosylation 0:14 N-Glycosylation 0:14 N-Glycosylation of the archaellum filament 0:41 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 6 34 theme N-Glycosylation 959:973 arg1 sites 975:979 the first five N-Glycosylation sites 944:979 the first five N-Glycosylation sites in FlaB 944:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 6 35 from FlaB 984:987 arg1 Deletion 932:939 Deletion 932:939 Deletion of the first five N-Glycosylation sites in FlaB 932:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 7 36 from change 1234:1239 arg1 length 1244:1249 length 1244:1249 length 1244:1249 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 7 37 gly glycosylated 1161:1172 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 1 38 theme modifications 238:250 arg1 one 197:199 one 197:199 one 197:199 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 1 38 theme modifications 238:250 arg1 modifications 238:250 the predominant posttranslational modifications 204:250 the predominant posttranslational modifications 204:250 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 0 39 theme Sulfolobus 151:160 arg1 acidocaldarius 162:175 Sulfolobus acidocaldarius 151:175 Sulfolobus acidocaldarius 151:175 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 4 40 theme predicted 559:567 arg1 sites 585:589 the six predicted N-Glycosylation sites 551:589 the six predicted N-Glycosylation sites within FlaB 551:601 Each of the six predicted N-Glycosylation sites within FlaB are modified with the attachment of an N-glycan. 25447136 1 41 located found 262:266 arg2 modifications 238:250 the predominant posttranslational modifications 204:250 the predominant posttranslational modifications 204:250 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 1 41 located found 262:266 arg1 life 292:295 life 292:295 life 292:295 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 1 41 located found 262:266 arg1 domains 281:287 all three domains 271:287 all three domains of life 271:295 N-Glycosylation is one of the predominant posttranslational modifications, which is found in all three domains of life. 25447136 7 42 theme non 1157:1159 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 6 43 theme first 948:952 arg1 sites 975:979 the first five N-Glycosylation sites 944:979 the first five N-Glycosylation sites in FlaB 944:987 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 8 44 theme archaellum 1415:1424 arg1 assembly 1426:1433 the archaellum assembly 1411:1433 the archaellum assembly 1411:1433 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 2 45 theme biological 347:356 arg1 aspects 358:364 many biological aspects 342:364 many biological aspects of proteins 342:376 N-Glycosylation has been shown to influence many biological aspects of proteins, like protein folding, stability or activity. 25447136 7 46 theme glycosylated 1178:1189 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 2 47 theme many 342:345 arg1 aspects 358:364 many biological aspects 342:364 many biological aspects of proteins 342:376 N-Glycosylation has been shown to influence many biological aspects of proteins, like protein folding, stability or activity. 25447136 0 48 from motility 139:146 arg1 acidocaldarius 162:175 Sulfolobus acidocaldarius 151:175 Sulfolobus acidocaldarius 151:175 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 3 49 theme subunit 482:488 arg1 N-glycosylated 527:540 N-glycosylated 527:540 N-glycosylated 527:540 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 3 49 theme subunit 482:488 arg1 FlaB 490:493 the archaellum filament subunit FlaB 458:493 the archaellum filament subunit FlaB of Sulfolobus acidocaldarius 458:522 In this study we demonstrate that the archaellum filament subunit FlaB of Sulfolobus acidocaldarius is N-glycosylated. 25447136 0 50 theme archaella 64:72 arg1 assembly 74:81 archaella assembly 64:81 archaella assembly 64:81 N-Glycosylation of the archaellum filament is not important for archaella assembly and motility, although N-Glycosylation is essential for motility in Sulfolobus acidocaldarius. 25447136 6 51 theme N-Glycosylation 1059:1073 arg1 sites 1075:1079 all six N-Glycosylation sites 1051:1079 all six N-Glycosylation sites 1051:1079 Deletion of the first five N-Glycosylation sites in FlaB did not significantly affect the motility, whereas removal of all six N-Glycosylation sites reduced motility by about 40%. 25447136 8 52 theme archaellum 1348:1357 arg1 filament 1359:1366 the archaellum filament itself 1344:1373 the archaellum filament itself 1344:1373 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 8 53 dep stability 1318:1326 arg1 the 1314:1316 the 1314:1316 the 1314:1316 Therefore N-Glycosylation does not appear to be important for the stability and assembly of the archaellum filament itself, but plays a role in other parts of the archaellum assembly. 25447136 7 54 gly glycosylated 1178:1189 arg1 filament 1202:1209 non glycosylated and glycosylated archaellum filament 1157:1209 non glycosylated and glycosylated archaellum filament 1157:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 25447136 7 55 theme filament 1202:1209 arg1 analyses 1145:1152 Transmission electron microscopy analyses 1112:1152 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament 1112:1209 Transmission electron microscopy analyses of non glycosylated and glycosylated archaellum filament revealed no structural change in length. 28525772 1 0 theme GlyTouCan 309:317 arg1 project 319:325 the GlyTouCan project 305:325 the GlyTouCan project 305:325 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 14 1 theme glycan 2028:2033 arg1 information 2035:2045 glycan information 2028:2045 glycan information 2028:2045 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 12 2 theme diverse 1761:1767 arg1 structures 1784:1793 the complex, diverse, and ambiguous structures 1748:1793 the complex, diverse, and ambiguous structures of glycans 1748:1804 This tool can aid researchers in communicating about the complex, diverse, and ambiguous structures of glycans more rapidly. 28525772 1 3 theme glycan 141:146 arg1 version 87:93 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 1 3 theme glycan 141:146 arg1 repository 158:167 the international glycan structure repository 123:167 the international glycan structure repository 123:167 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 0 4 theme glycans 68:74 arg1 glycans 68:74 ambiguous glycans 58:74 ambiguous glycans 58:74 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 0 4 theme glycans 68:74 arg1 variety 47:53 a wide variety 40:53 a wide variety of ambiguous glycans 40:74 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 1 5 theme project 319:325 arg1 stages 295:300 the early stages 285:300 the early stages of the GlyTouCan project 285:325 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 1 6 theme structure 148:156 arg1 version 87:93 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 1 6 theme structure 148:156 arg1 repository 158:167 the international glycan structure repository 123:167 the international glycan structure repository 123:167 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 10 7 theme GlycanBuilder 1462:1474 arg1 version 1451:1457 This new version 1442:1457 This new version 1442:1457 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 8 theme ambiguous 1538:1546 arg1 glycans 1548:1554 ambiguous glycans 1538:1554 ambiguous glycans 1538:1554 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 14 9 theme glycans 2067:2073 arg1 glycans 2067:2073 glycans 2067:2073 glycans 2067:2073 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 9 theme glycans 2067:2073 arg1 variety 2056:2062 a wider variety 2048:2062 a wider variety of glycans 2048:2073 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 10 theme basic 1981:1985 arg1 format 1987:1992 the basic format 1977:1992 the basic format for registration 1977:2009 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 10 theme basic 1981:1985 arg1 WURCS 1968:1972 WURCS 1968:1972 WURCS 1968:1972 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 1 11 theme Carbohydrate 240:251 arg1 Structures 253:262 Carbohydrate Structures 240:262 Carbohydrate Structures 240:262 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 3 12 theme existing 546:553 arg1 structures 569:578 existing or new glycan structures 546:578 existing or new glycan structures 546:578 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 3 13 theme WURCS 528:532 arg1 strings 534:540 WURCS strings 528:540 WURCS strings for existing or new glycan structures 528:578 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 1 14 theme Structures 253:262 arg1 Representation 222:235 Web3 Unique Representation 210:235 Web3 Unique Representation of Carbohydrate Structures 210:262 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 12 15 theme ambiguous 1774:1782 arg1 structures 1784:1793 the complex, diverse, and ambiguous structures 1748:1793 the complex, diverse, and ambiguous structures of glycans 1748:1804 This tool can aid researchers in communicating about the complex, diverse, and ambiguous structures of glycans more rapidly. 28525772 2 16 from flexibility 440:450 arg1 insufficient 418:429 insufficient 418:429 insufficient 418:429 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 2 17 theme existing 396:403 arg1 formats 405:411 existing formats 396:411 existing formats 396:411 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 9 18 theme symbol 1421:1426 arg1 nomenclature 1428:1439 the latest glycan symbol nomenclature 1403:1439 the latest glycan symbol nomenclature 1403:1439 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 8 19 theme glycan 1215:1220 arg1 structures 1222:1231 cross-linked glycan structures 1202:1231 cross-linked glycan structures 1202:1231 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 8 20 theme cyclic 1130:1135 arg1 glycans 1137:1143 cyclic glycans 1130:1143 cyclic glycans 1130:1143 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 10 21 from bacteria 1610:1617 arg1 monosaccharides 1589:1603 monosaccharides 1589:1603 monosaccharides from bacteria and plants 1589:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 22 theme new 1447:1449 arg1 version 1451:1457 This new version 1442:1457 This new version 1442:1457 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 5 23 theme mammalian 834:842 arg1 glycans 844:850 mammalian glycans 834:850 mammalian glycans 834:850 However, GlycanBuilder was limited because it was originally developed to support mammalian glycans. 28525772 8 24 theme nested 1146:1151 arg1 units 1163:1167 nested repeating units 1146:1167 nested repeating units 1146:1167 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 6 25 theme Symbol 934:939 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 1 26 theme GlyTouCan 77:85 arg1 version 87:93 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 1 26 theme GlyTouCan 77:85 arg1 repository 158:167 the international glycan structure repository 123:167 the international glycan structure repository 123:167 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 12 27 theme glycans 1798:1804 arg1 structures 1784:1793 the complex, diverse, and ambiguous structures 1748:1793 the complex, diverse, and ambiguous structures of glycans 1748:1804 This tool can aid researchers in communicating about the complex, diverse, and ambiguous structures of glycans more rapidly. 28525772 2 28 theme base 356:359 arg1 representation 361:374 its base representation 352:374 its base representation for glycans 352:386 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 9 29 theme export 1269:1274 arg1 utilities 1276:1284 Both import and export utilities 1253:1284 Both import and export utilities for WURCS 1253:1294 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 9 30 theme import 1258:1263 arg1 utilities 1276:1284 Both import and export utilities 1253:1284 Both import and export utilities for WURCS 1253:1294 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 11 31 theme new 1677:1679 arg1 symbols 1686:1692 the new SNFG symbols 1673:1692 the new SNFG symbols 1673:1692 These glycans can also be displayed using the new SNFG symbols. 28525772 14 32 theme information 2035:2045 arg1 format 1987:1992 the basic format 1977:1992 the basic format for registration 1977:2009 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 32 theme information 2035:2045 arg1 WURCS 1968:1972 WURCS 1968:1972 WURCS 1968:1972 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 32 theme information 2035:2045 arg1 searching 2015:2023 searching 2015:2023 searching of glycan information 2015:2045 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 4 33 theme wide 723:726 arg1 usage 728:732 its wide usage 719:732 its wide usage by the community 719:749 GlycanBuilder was an obvious choice to extend due to its wide usage by the community. 28525772 13 34 theme new 1834:1836 arg1 GlycanBuilder 1838:1850 the new GlycanBuilder 1830:1850 the new GlycanBuilder 1830:1850 Moreover, the new GlycanBuilder can now easily output WURCS sequences from glycans drawn on the canvas. 28525772 1 35 theme new 176:178 arg1 format 189:194 a new sequence format 174:194 a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) 174:263 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 0 36 theme GlycanBuilder 18:30 arg1 Implementation 0:13 Implementation 0:13 Implementation of GlycanBuilder 0:30 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 9 37 theme SNFG 1322:1325 arg1 symbols 1327:1333 SNFG symbols 1322:1333 SNFG symbols 1322:1333 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 7 38 theme GlycanBuilder 1030:1042 arg1 version 1019:1025 a new version 1013:1025 a new version of GlycanBuilder 1013:1042 Therefore in this work, we implemented a new version of GlycanBuilder to greatly increase its usability. 28525772 1 39 theme sequence 180:187 arg1 format 189:194 a new sequence format 174:194 a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) 174:263 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 8 40 theme monosaccharide 1170:1183 arg1 compositions 1185:1196 monosaccharide compositions 1170:1196 monosaccharide compositions 1170:1196 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 4 41 theme obvious 687:693 arg1 choice 695:700 an obvious choice to extend due to its wide usage by the community 684:749 an obvious choice to extend due to its wide usage by the community 684:749 GlycanBuilder was an obvious choice to extend due to its wide usage by the community. 28525772 4 41 theme obvious 687:693 arg1 GlycanBuilder 666:678 GlycanBuilder 666:678 GlycanBuilder 666:678 GlycanBuilder was an obvious choice to extend due to its wide usage by the community. 28525772 7 42 theme new 1015:1017 arg1 version 1019:1025 a new version 1013:1025 a new version of GlycanBuilder 1013:1042 Therefore in this work, we implemented a new version of GlycanBuilder to greatly increase its usability. 28525772 10 43 theme glycans 1548:1554 arg1 variety 1527:1533 a wide variety 1520:1533 a wide variety 1520:1533 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 43 theme glycans 1548:1554 arg1 glycans 1548:1554 ambiguous glycans 1538:1554 ambiguous glycans 1538:1554 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 43 theme glycans 1548:1554 arg1 structures 1567:1576 structures 1567:1576 structures containing monosaccharides from bacteria and plants 1567:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 9 44 theme latest 1407:1412 arg1 nomenclature 1428:1439 the latest glycan symbol nomenclature 1403:1439 the latest glycan symbol nomenclature 1403:1439 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 6 45 theme called 927:932 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 1 46 dep WURCS 203:207 arg1 Representation 222:235 Web3 Unique Representation 210:235 Web3 Unique Representation of Carbohydrate Structures 210:262 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 10 47 from plants 1623:1628 arg1 monosaccharides 1589:1603 monosaccharides 1589:1603 monosaccharides from bacteria and plants 1589:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 48 theme wide 1522:1525 arg1 variety 1527:1533 a wide variety 1520:1533 a wide variety 1520:1533 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 48 theme wide 1522:1525 arg1 glycans 1548:1554 ambiguous glycans 1538:1554 ambiguous glycans 1538:1554 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 48 theme wide 1522:1525 arg1 structures 1567:1576 structures 1567:1576 structures containing monosaccharides from bacteria and plants 1567:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 1 49 theme early 289:293 arg1 stages 295:300 the early stages 285:300 the early stages of the GlyTouCan project 285:325 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 2 50 from glycans 477:483 arg1 insufficient 418:429 insufficient 418:429 insufficient 418:429 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 3 51 theme new 558:560 arg1 structures 569:578 existing or new glycan structures 546:578 existing or new glycan structures 546:578 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 0 52 theme wide 42:45 arg1 glycans 68:74 ambiguous glycans 58:74 ambiguous glycans 58:74 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 0 52 theme wide 42:45 arg1 variety 47:53 a wide variety 40:53 a wide variety of ambiguous glycans 40:74 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 6 53 theme symbol 911:916 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 6 54 theme proposed 887:894 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 3 55 theme glycan 562:567 arg1 structures 569:578 existing or new glycan structures 546:578 existing or new glycan structures 546:578 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 3 56 theme structure 608:616 arg1 editors 618:624 glycan structure editors 601:624 glycan structure editors 601:624 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 13 57 theme output 1867:1872 arg1 sequences 1880:1888 output WURCS sequences 1867:1888 output WURCS sequences from glycans drawn on the canvas 1867:1921 Moreover, the new GlycanBuilder can now easily output WURCS sequences from glycans drawn on the canvas. 28525772 6 58 theme monosaccharide 896:909 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 8 59 theme glycan 1083:1088 arg1 system 1100:1105 The glycan rendering system 1079:1105 The glycan rendering system 1079:1105 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 10 60 contain containing 1578:1587 arg2 monosaccharides 1589:1603 monosaccharides 1589:1603 monosaccharides from bacteria and plants 1589:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 10 60 contain containing 1578:1587 arg1 structures 1567:1576 structures 1567:1576 structures containing monosaccharides from bacteria and plants 1567:1628 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 8 61 link cross-linked 1202:1213 arg1 structures 1222:1231 cross-linked glycan structures 1202:1231 cross-linked glycan structures 1202:1231 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 13 62 from glycans 1895:1901 arg1 sequences 1880:1888 output WURCS sequences 1867:1888 output WURCS sequences from glycans drawn on the canvas 1867:1921 Moreover, the new GlycanBuilder can now easily output WURCS sequences from glycans drawn on the canvas. 28525772 13 63 theme WURCS 1874:1878 arg1 sequences 1880:1888 output WURCS sequences 1867:1888 output WURCS sequences from glycans drawn on the canvas 1867:1921 Moreover, the new GlycanBuilder can now easily output WURCS sequences from glycans drawn on the canvas. 28525772 9 64 theme glycan 1414:1419 arg1 nomenclature 1428:1439 the latest glycan symbol nomenclature 1403:1439 the latest glycan symbol nomenclature 1403:1439 Both import and export utilities for WURCS were also implemented and SNFG symbols were incorporated to allow glycans to be exported as graphics using the latest glycan symbol nomenclature. 28525772 10 65 theme GlycanBuilder2 1484:1497 arg1 " 1498:1498 GlycanBuilder2" 1484:1498 GlycanBuilder2" 1484:1498 This new version of GlycanBuilder called "GlycanBuilder2", is able to support a wide variety of ambiguous glycans, including structures containing monosaccharides from bacteria and plants. 28525772 11 66 theme SNFG 1681:1684 arg1 symbols 1686:1692 the new SNFG symbols 1673:1692 the new SNFG symbols 1673:1692 These glycans can also be displayed using the new SNFG symbols. 28525772 3 67 theme glycan 601:606 arg1 editors 618:624 glycan structure editors 601:624 glycan structure editors 601:624 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 14 68 theme wider 2050:2054 arg1 glycans 2067:2073 glycans 2067:2073 glycans 2067:2073 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 14 68 theme wider 2050:2054 arg1 variety 2056:2062 a wider variety 2048:2062 a wider variety of glycans 2048:2073 Most importantly, because GlyTouCan employs WURCS as the basic format for registration and searching of glycan information, a wider variety of glycans can now be readily registered and queried in GlyTouCan. 28525772 0 69 theme ambiguous 58:66 arg1 glycans 68:74 ambiguous glycans 58:74 ambiguous glycans 58:74 Implementation of GlycanBuilder to draw a wide variety of ambiguous glycans. 28525772 3 70 theme conversion 581:590 arg1 tools 592:596 conversion tools 581:596 conversion tools 581:596 Therefore, in order to obtain WURCS strings for existing or new glycan structures, conversion tools or glycan structure editors that can export WURCS became necessary. 28525772 12 71 theme complex 1752:1758 arg1 structures 1784:1793 the complex, diverse, and ambiguous structures 1748:1793 the complex, diverse, and ambiguous structures of glycans 1748:1804 This tool can aid researchers in communicating about the complex, diverse, and ambiguous structures of glycans more rapidly. 28525772 8 72 theme cross-linked 1202:1213 arg1 structures 1222:1231 cross-linked glycan structures 1202:1231 cross-linked glycan structures 1202:1231 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 1 73 theme Web3 210:213 arg1 Representation 222:235 Web3 Unique Representation 210:235 Web3 Unique Representation of Carbohydrate Structures 210:262 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 8 74 theme rendering 1090:1098 arg1 system 1100:1105 The glycan rendering system 1079:1105 The glycan rendering system 1079:1105 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 2 75 from insufficient 418:429 arg1 glycans 477:483 all glycans 473:483 all glycans universally 473:495 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 2 75 from insufficient 418:429 arg1 flexibility 440:450 their flexibility to represent any 434:467 their flexibility to represent any 434:467 GlyTouCan uses WURCS as its base representation for glycans because existing formats were insufficient in their flexibility to represent any and all glycans universally. 28525772 8 76 theme repeating 1153:1161 arg1 units 1163:1167 nested repeating units 1146:1167 nested repeating units 1146:1167 The glycan rendering system was refactored so that cyclic glycans, nested repeating units, monosaccharide compositions and cross-linked glycan structures can be represented. 28525772 1 77 theme international 127:139 arg1 version 87:93 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 77:97 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 1 77 theme international 127:139 arg1 repository 158:167 the international glycan structure repository 123:167 the international glycan structure repository 123:167 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 28525772 6 78 theme standard 918:925 arg1 Nomenclature 941:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature 877:952 the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG) 877:971 It also did not support the newly proposed monosaccharide symbol standard called Symbol Nomenclature for Glycans (SNFG). 28525772 1 79 theme Unique 215:220 arg1 Representation 222:235 Web3 Unique Representation 210:235 Web3 Unique Representation of Carbohydrate Structures 210:262 GlyTouCan version 1.0 was released in 2015 as the international glycan structure repository, and a new sequence format called WURCS (Web3 Unique Representation of Carbohydrate Structures) was proposed during the early stages of the GlyTouCan project. 27428423 6 0 theme set 1197:1199 arg1 regulation 1174:1183 androgen regulation 1165:1183 androgen regulation of the core set of enzymes 1165:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 1 theme glycan 1234:1239 arg1 synthesis 1241:1249 glycan synthesis 1234:1249 glycan synthesis 1234:1249 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 2 theme sialyl 1093:1098 arg1 Lewis 1100:1104 sialyl Lewis 1093:1104 sialyl Lewis(X) (SLe(X)) 1093:1116 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 2 theme sialyl 1093:1098 arg1 X 1106:1106 X 1106:1106 X 1106:1106 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 2 theme sialyl 1093:1098 arg1 SLe 1110:1112 SLe 1110:1112 SLe(X) 1110:1115 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 4 3 from enrichment 709:718 arg1 glycosylation 744:756 the cellular process glycosylation 723:756 the cellular process glycosylation 723:756 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 6 4 theme downstream 1017:1026 arg1 structures 1006:1015 glycan structures 999:1015 glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 999:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 8 5 theme specific 1438:1445 arg1 enzymes 1461:1467 specific glycosylation enzymes 1438:1467 specific glycosylation enzymes 1438:1467 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 3 6 from expression 499:508 arg1 response 513:520 response 513:520 response to acute androgen stimulation in culture 513:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 2 7 theme cellular 353:360 arg1 pathways 362:369 androgen-regulated cellular pathways 334:369 androgen-regulated cellular pathways that may be clinically important in prostate cancer 334:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 3 8 from stimulation 540:550 arg1 culture 555:561 culture 555:561 culture 555:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 5 9 theme clinical 928:935 arg1 carcinoma 946:954 clinical prostate carcinoma 928:954 clinical prostate carcinoma 928:954 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 8 10 theme enzymes 1461:1467 arg1 loss 1430:1433 loss 1430:1433 loss of specific glycosylation enzymes 1430:1467 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 6 11 theme chondroitin 1132:1142 arg1 sulphate 1144:1151 chondroitin sulphate 1132:1151 chondroitin sulphate 1132:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 12 theme regulated 1048:1056 arg1 enzymes 1058:1064 regulated enzymes 1048:1064 regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 1048:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 12 theme regulated 1048:1056 arg1 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 12 theme regulated 1048:1056 arg1 sulphate 1144:1151 chondroitin sulphate 1132:1151 chondroitin sulphate 1132:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 12 theme regulated 1048:1056 arg1 Lewis 1100:1104 sialyl Lewis 1093:1104 sialyl Lewis(X) (SLe(X)) 1093:1116 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 12 theme regulated 1048:1056 arg1 sialyl-Tn 1076:1084 sialyl-Tn 1076:1084 sialyl-Tn (sTn) 1076:1090 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 13 theme glycan 999:1004 arg1 structures 1006:1015 glycan structures 999:1015 glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 999:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 2 14 theme search 306:311 arg1 strategy 313:320 a novel search strategy 298:320 a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer 298:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 6 15 theme key 1221:1223 arg1 steps 1225:1229 key steps 1221:1229 key steps 1221:1229 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 1 16 theme cancer 269:274 arg1 growth 276:281 cancer growth 269:281 cancer growth 269:281 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 3 17 with patients 592:599 arg1 cancer 615:620 prostate cancer 606:620 prostate cancer 606:620 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 4 18 theme cellular 727:734 arg1 glycosylation 744:756 the cellular process glycosylation 723:756 the cellular process glycosylation 723:756 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 5 19 dep 8 820:820 arg1 GCNT1 843:847 GCNT1 843:847 GCNT1 843:847 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 UAP1 850:853 UAP1 850:853 UAP1 850:853 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 ST6GAL1 874:880 ST6GAL1 874:880 ST6GAL1 874:880 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 GALNT7 823:828 GALNT7 823:828 GALNT7 823:828 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 EDEM3 886:890 EDEM3 886:890 EDEM3 886:890 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 ST6GalNAc1 831:840 ST6GalNAc1 831:840 ST6GalNAc1 831:840 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 PGM3 856:859 PGM3 856:859 PGM3 856:859 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 19 dep 8 820:820 arg1 CSGALNACT1 862:871 CSGALNACT1 862:871 CSGALNACT1 862:871 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 4 20 theme 700 631:633 arg1 genes 635:639 700 genes 631:639 700 genes displaying reciprocal expression patterns 631:681 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 3 21 theme acute 525:529 arg1 stimulation 540:550 acute androgen stimulation 525:550 acute androgen stimulation in culture 525:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 3 22 theme RNASeq 430:435 arg1 data 437:440 RNASeq data 430:440 RNASeq data 430:440 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 6 23 theme enzymes 1058:1064 arg1 enzymes 1058:1064 regulated enzymes 1048:1064 regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 1048:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 23 theme enzymes 1058:1064 arg1 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 23 theme enzymes 1058:1064 arg1 Lewis 1100:1104 sialyl Lewis 1093:1104 sialyl Lewis(X) (SLe(X)) 1093:1116 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 23 theme enzymes 1058:1064 arg1 sialyl-Tn 1076:1084 sialyl-Tn 1076:1084 sialyl-Tn (sTn) 1076:1090 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 23 theme enzymes 1058:1064 arg1 set 1041:1043 this core set 1031:1043 this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 1031:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 23 theme enzymes 1058:1064 arg1 sulphate 1144:1151 chondroitin sulphate 1132:1151 chondroitin sulphate 1132:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 5 24 theme reciprocally-regulated 765:786 arg1 enzymes 802:808 31 reciprocally-regulated glycosylation enzymes 762:808 31 reciprocally-regulated glycosylation enzymes 762:808 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 6 25 theme enzymes 1204:1210 arg1 enzymes 1204:1210 enzymes 1204:1210 enzymes 1204:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 25 theme enzymes 1204:1210 arg1 set 1197:1199 the core set 1188:1199 the core set of enzymes 1188:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 3 26 theme androgen 531:538 arg1 stimulation 540:550 acute androgen stimulation 525:550 acute androgen stimulation in culture 525:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 1 27 theme prostate 171:178 arg1 cancer 180:185 prostate cancer 171:185 prostate cancer 171:185 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 8 28 theme tumour 1489:1494 arg1 regression 1496:1505 tumour regression 1489:1505 tumour regression following androgen depletion therapy 1489:1542 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 5 29 theme glycosylation 788:800 arg1 enzymes 802:808 31 reciprocally-regulated glycosylation enzymes 762:808 31 reciprocally-regulated glycosylation enzymes 762:808 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 1 30 theme Steroid 93:99 arg1 hormones 110:117 Steroid androgen hormones 93:117 Steroid androgen hormones 93:117 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 31 theme cancer 180:185 arg1 progression 142:152 progression 142:152 progression 142:152 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 31 theme cancer 180:185 arg1 treatment 158:166 treatment 158:166 treatment 158:166 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 8 32 theme androgen 1517:1524 arg1 therapy 1536:1542 androgen depletion therapy 1517:1542 androgen depletion therapy 1517:1542 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 4 33 theme process 736:742 arg1 glycosylation 744:756 the cellular process glycosylation 723:756 the cellular process glycosylation 723:756 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 3 34 from changes 488:494 arg1 patients 592:599 patients 592:599 patients with prostate cancer 592:620 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 3 34 from changes 488:494 arg1 expression 499:508 expression 499:508 expression in response to acute androgen stimulation in culture 499:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 1 35 theme androgen 101:108 arg1 hormones 110:117 Steroid androgen hormones 93:117 Steroid androgen hormones 93:117 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 3 36 from deprivation 577:587 arg1 patients 592:599 patients 592:599 patients with prostate cancer 592:620 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 3 36 from deprivation 577:587 arg1 expression 499:508 expression 499:508 expression in response to acute androgen stimulation in culture 499:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 0 37 theme Androgen-Regulated 20:37 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation 0:12 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 0 37 theme Androgen-Regulated 20:37 arg1 Process 39:45 an Androgen-Regulated Process 17:45 an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability 17:90 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 5 38 theme 8 820:820 arg1 8 820:820 8 820:820 8 820:820 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 5 38 theme 8 820:820 arg1 set 813:815 a set 811:815 a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) 811:891 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 2 39 theme novel 300:304 arg1 strategy 313:320 a novel search strategy 298:320 a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer 298:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 5 40 theme prostate 937:944 arg1 carcinoma 946:954 clinical prostate carcinoma 928:954 clinical prostate carcinoma 928:954 Of 31 reciprocally-regulated glycosylation enzymes, a set of 8 (GALNT7, ST6GalNAc1, GCNT1, UAP1, PGM3, CSGALNACT1, ST6GAL1 and EDEM3) were significantly up-regulated in clinical prostate carcinoma. 27428423 8 41 theme androgen 1399:1406 arg1 control 1408:1414 androgen control 1399:1414 androgen control 1399:1414 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 1 42 dep progression 142:152 arg1 the 138:140 the 138:140 the 138:140 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 43 theme androgen 193:200 arg1 treatment 243:251 the first-line treatment 228:251 the first-line treatment used to control cancer growth 228:281 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 43 theme androgen 193:200 arg1 therapy 214:220 androgen deprivation therapy 193:220 androgen deprivation therapy 193:220 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 0 44 theme Essential 47:55 arg1 Glycosylation 0:12 Glycosylation 0:12 Glycosylation 0:12 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 0 44 theme Essential 47:55 arg1 Process 39:45 an Androgen-Regulated Process 17:45 an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability 17:90 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 6 45 theme Androgen 957:964 arg1 exposure 966:973 Androgen exposure 957:973 Androgen exposure 957:973 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 4 46 theme reciprocal 652:661 arg1 patterns 674:681 reciprocal expression patterns 652:681 reciprocal expression patterns 652:681 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 6 47 theme core 1192:1195 arg1 enzymes 1204:1210 enzymes 1204:1210 enzymes 1204:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 47 theme core 1192:1195 arg1 set 1197:1199 the core set 1188:1199 the core set of enzymes 1188:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 1 48 theme deprivation 202:212 arg1 treatment 243:251 the first-line treatment 228:251 the first-line treatment used to control cancer growth 228:281 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 48 theme deprivation 202:212 arg1 therapy 214:220 androgen deprivation therapy 193:220 androgen deprivation therapy 193:220 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 6 49 theme structures 1006:1015 arg1 synthesis 986:994 synthesis 986:994 synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 986:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 2 50 theme prostate 407:414 arg1 cancer 416:421 prostate cancer 407:421 prostate cancer 407:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 0 51 theme Prostate 61:68 arg1 Viability 82:90 Prostate Cancer Cell Viability 61:90 Prostate Cancer Cell Viability 61:90 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 7 52 theme cancer 1317:1322 arg1 viability 1329:1337 prostate cancer cell viability 1308:1337 prostate cancer cell viability 1308:1337 Expression of each of these enzymes also contributed to prostate cancer cell viability. 27428423 2 53 theme androgen-regulated 334:351 arg1 pathways 362:369 androgen-regulated cellular pathways 334:369 androgen-regulated cellular pathways that may be clinically important in prostate cancer 334:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 8 54 theme depletion 1526:1534 arg1 therapy 1536:1542 androgen depletion therapy 1517:1542 androgen depletion therapy 1517:1542 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 7 55 theme cell 1324:1327 arg1 viability 1329:1337 prostate cancer cell viability 1308:1337 prostate cancer cell viability 1308:1337 Expression of each of these enzymes also contributed to prostate cancer cell viability. 27428423 1 56 theme key 126:128 arg1 role 130:133 a key role 124:133 a key role 124:133 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 6 57 theme androgen 1165:1172 arg1 regulation 1174:1183 androgen regulation 1165:1183 androgen regulation of the core set of enzymes 1165:1210 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 2 58 from important 394:402 arg1 cancer 416:421 prostate cancer 407:421 prostate cancer 407:421 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 6 59 theme set 1041:1043 arg1 downstream 1017:1026 downstream 1017:1026 downstream 1017:1026 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 7 60 theme prostate 1308:1315 arg1 viability 1329:1337 prostate cancer cell viability 1308:1337 prostate cancer cell viability 1308:1337 Expression of each of these enzymes also contributed to prostate cancer cell viability. 27428423 2 61 from cancer 416:421 arg1 important 394:402 important 394:402 important 394:402 Here we apply a novel search strategy to identify androgen-regulated cellular pathways that may be clinically important in prostate cancer. 27428423 3 62 theme reciprocal 477:486 arg1 changes 488:494 reciprocal changes 477:494 reciprocal changes in expression in response to acute androgen stimulation in culture 477:561 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 7 63 theme each 1266:1269 arg1 Expression 1252:1261 Expression 1252:1261 Expression of each of these enzymes 1252:1286 Expression of each of these enzymes also contributed to prostate cancer cell viability. 27428423 6 64 theme core 1036:1039 arg1 enzymes 1058:1064 regulated enzymes 1048:1064 regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 1048:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 64 theme core 1036:1039 arg1 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 O-GlcNAc 1119:1126 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 64 theme core 1036:1039 arg1 Lewis 1100:1104 sialyl Lewis 1093:1104 sialyl Lewis(X) (SLe(X)) 1093:1116 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 64 theme core 1036:1039 arg1 sialyl-Tn 1076:1084 sialyl-Tn 1076:1084 sialyl-Tn (sTn) 1076:1090 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 64 theme core 1036:1039 arg1 set 1041:1043 this core set 1031:1043 this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate 1031:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 6 64 theme core 1036:1039 arg1 sulphate 1144:1151 chondroitin sulphate 1132:1151 chondroitin sulphate 1132:1151 Androgen exposure stimulated synthesis of glycan structures downstream of this core set of regulated enzymes including sialyl-Tn (sTn), sialyl Lewis(X) (SLe(X)), O-GlcNAc and chondroitin sulphate, suggesting androgen regulation of the core set of enzymes controls key steps in glycan synthesis. 27428423 0 65 theme Cell 77:80 arg1 Viability 82:90 Prostate Cancer Cell Viability 61:90 Prostate Cancer Cell Viability 61:90 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 3 66 theme prostate 606:613 arg1 cancer 615:620 prostate cancer 606:620 prostate cancer 606:620 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27428423 4 67 theme significant 697:707 arg1 enrichment 709:718 a significant enrichment 695:718 a significant enrichment in the cellular process glycosylation 695:756 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 4 68 theme expression 663:672 arg1 patterns 674:681 reciprocal expression patterns 652:681 reciprocal expression patterns 652:681 Amongst 700 genes displaying reciprocal expression patterns we observed a significant enrichment in the cellular process glycosylation. 27428423 1 69 theme first-line 232:241 arg1 treatment 243:251 the first-line treatment 228:251 the first-line treatment used to control cancer growth 228:281 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 1 69 theme first-line 232:241 arg1 therapy 214:220 androgen deprivation therapy 193:220 androgen deprivation therapy 193:220 Steroid androgen hormones play a key role in the progression and treatment of prostate cancer, with androgen deprivation therapy being the first-line treatment used to control cancer growth. 27428423 0 70 theme Cancer 70:75 arg1 Viability 82:90 Prostate Cancer Cell Viability 61:90 Prostate Cancer Cell Viability 61:90 Glycosylation is an Androgen-Regulated Process Essential for Prostate Cancer Cell Viability. 27428423 8 71 theme glycosylation 1447:1459 arg1 enzymes 1461:1467 specific glycosylation enzymes 1438:1467 specific glycosylation enzymes 1438:1467 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 8 72 theme global 1381:1386 arg1 glycosylation 1362:1374 glycosylation 1362:1374 glycosylation 1362:1374 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 8 72 theme global 1381:1386 arg1 target 1388:1393 a global target 1379:1393 a global target for androgen control 1379:1414 This study identifies glycosylation as a global target for androgen control, and suggests loss of specific glycosylation enzymes might contribute to tumour regression following androgen depletion therapy. 27428423 3 73 theme androgen 568:575 arg1 deprivation 577:587 androgen deprivation 568:587 androgen deprivation in patients with prostate cancer 568:620 Using RNASeq data, we searched for genes that showed reciprocal changes in expression in response to acute androgen stimulation in culture, and androgen deprivation in patients with prostate cancer. 27994047 8 0 dep large 1302:1306 arg1 kDa 1315:1317 >2500 kDa 1309:1317 >2500 kDa 1309:1317 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 0 dep large 1302:1306 arg1 glycosylated 1328:1339 glycosylated 1328:1339 glycosylated 1328:1339 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 11 1 theme inaccurate 1802:1811 arg1 conclusions 1813:1823 inaccurate conclusions 1802:1823 inaccurate conclusions 1802:1823 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 0 2 theme proteoglycans 73:85 arg1 preparations 27:38 commercial preparations 16:38 commercial preparations of 'purified' small leucine-rich proteoglycans 16:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 4 3 theme leucine-rich 539:550 arg1 SLRPs 567:571 SLRPs 567:571 SLRPs 567:571 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 3 theme leucine-rich 539:550 arg1 proteoglycans 552:564 the small leucine-rich proteoglycans 529:564 the small leucine-rich proteoglycans (SLRPs) 529:572 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 0 4 theme mechanistic 99:109 arg1 studies 111:117 mechanistic studies 99:117 mechanistic studies 99:117 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 4 5 theme small 533:537 arg1 SLRPs 567:571 SLRPs 567:571 SLRPs 567:571 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 5 theme small 533:537 arg1 proteoglycans 552:564 the small leucine-rich proteoglycans 529:564 the small leucine-rich proteoglycans (SLRPs) 529:572 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 8 6 theme chondroitin 1376:1386 arg1 sulfate 1388:1394 chondroitin sulfate 1376:1394 chondroitin sulfate 1376:1394 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 7 7 from effects 1047:1053 arg1 growth 1097:1102 nerve cell growth 1086:1102 nerve cell growth 1086:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 5 8 theme proteoglycans 747:759 arg1 preparations 725:736 both preparations 720:736 both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting 720:820 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 7 9 theme biglycan 1073:1080 arg1 effects 1047:1053 The unexpected effects 1032:1053 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth 1032:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 8 10 theme large 1302:1306 arg1 aggrecan 1341:1348 the large (>2500 kDa), highly glycosylated aggrecan 1298:1348 the large (>2500 kDa), highly glycosylated aggrecan 1298:1348 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 10 11 theme obtained 1651:1658 arg1 reagent 1660:1666 each commercially obtained reagent 1633:1666 each commercially obtained reagent used in their experiments 1633:1692 These findings beg the question as to whether scientists need to verify the purity of each commercially obtained reagent used in their experiments. 27994047 4 12 from influence 618:626 arg1 growth 642:647 nerve cell growth 631:647 nerve cell growth 631:647 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 5 13 dep composition 694:704 arg1 the 690:692 the 690:692 the 690:692 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 5 14 theme enzymatic 848:856 arg1 deglycosylations 858:873 various enzymatic deglycosylations 840:873 various enzymatic deglycosylations 840:873 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 0 15 theme purified 44:51 arg1 proteoglycans 73:85 'purified' small leucine-rich proteoglycans 43:85 'purified' small leucine-rich proteoglycans 43:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 11 16 theme findings 1721:1728 arg1 vast 1734:1737 vast 1734:1737 vast 1734:1737 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 11 16 theme findings 1721:1728 arg1 implications 1699:1710 The implications 1695:1710 The implications of these findings 1695:1728 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 9 17 theme different 1401:1409 arg1 structure 1411:1419 different structure 1401:1419 different structure 1401:1419 The different structure, molecular weight and composition of these impurities significantly affected our work and any conclusions that could be made. 27994047 4 18 theme sourced 505:511 arg1 decorin 575:581 decorin 575:581 decorin 575:581 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 18 theme sourced 505:511 arg1 preparations 513:524 commercially sourced preparations 492:524 commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs) 492:572 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 18 theme sourced 505:511 arg1 biglycan 587:594 biglycan 587:594 biglycan 587:594 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 5 19 theme Unusual 650:656 arg1 results 658:664 Unusual results 650:664 Unusual results 650:664 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 8 20 contain contains 1350:1357 arg1 aggrecan 1341:1348 the large (>2500 kDa), highly glycosylated aggrecan 1298:1348 the large (>2500 kDa), highly glycosylated aggrecan 1298:1348 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 20 contain contains 1350:1357 arg2 sulfate 1388:1394 chondroitin sulfate 1376:1394 chondroitin sulfate 1376:1394 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 20 contain contains 1350:1357 arg2 keratan 1364:1370 keratan 1364:1370 keratan 1364:1370 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 21 theme glycosaminoglycan 1212:1228 arg1 chains 1236:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 5 22 theme Western 805:811 arg1 blotting 813:820 Western blotting 805:820 Western blotting 805:820 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 7 23 theme decorin 1059:1065 arg1 effects 1047:1053 The unexpected effects 1032:1053 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth 1032:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 8 24 theme sulfate 1204:1210 arg1 glycosaminoglycan 1212:1228 either chondroitin or dermatan sulfate glycosaminoglycan 1173:1228 glycosaminoglycan 1212:1228 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 6 25 theme PGs 947:949 arg1 mixture 936:942 a mixture 934:942 a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan 934:1029 Commercial 'decorin' and 'biglycan' were found to contain a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan. 27994047 9 26 theme molecular 1422:1430 arg1 weight 1432:1437 molecular weight 1422:1437 molecular weight 1422:1437 The different structure, molecular weight and composition of these impurities significantly affected our work and any conclusions that could be made. 27994047 0 27 theme commercial 16:25 arg1 preparations 27:38 commercial preparations 16:38 commercial preparations of 'purified' small leucine-rich proteoglycans 16:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 6 28 theme Commercial 876:885 arg1 decorin 888:894 Commercial 'decorin' 876:895 Commercial 'decorin' 876:895 Commercial 'decorin' and 'biglycan' were found to contain a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan. 27994047 1 29 theme impure 206:211 arg1 reagents 236:243 seriously impure commercially available reagents 196:243 seriously impure commercially available reagents 196:243 The present study reports the perplexing results that came about because of seriously impure commercially available reagents. 27994047 5 30 theme preparations 725:736 arg1 purity 710:715 purity 710:715 purity 710:715 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 5 30 theme preparations 725:736 arg1 composition 694:704 composition 694:704 composition 694:704 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 8 31 theme dermatan 1195:1202 arg1 glycosaminoglycan 1212:1228 either chondroitin or dermatan sulfate glycosaminoglycan 1173:1228 glycosaminoglycan 1212:1228 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 5 32 theme mass 778:781 arg1 MS 797:798 MS 797:798 MS 797:798 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 5 32 theme mass 778:781 arg1 spectrometry 783:794 mass spectrometry 778:794 mass spectrometry (MS) 778:799 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 2 33 theme Commercial 246:255 arg1 reagents 257:264 Commercial reagents 246:264 Commercial reagents 246:264 Commercial reagents and chemicals are routinely ordered by scientists and expected to have been rigorously assessed for their purity. 27994047 1 34 theme available 226:234 arg1 reagents 236:243 seriously impure commercially available reagents 196:243 seriously impure commercially available reagents 196:243 The present study reports the perplexing results that came about because of seriously impure commercially available reagents. 27994047 7 35 theme nerve 1086:1090 arg1 growth 1097:1102 nerve cell growth 1086:1102 nerve cell growth 1086:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 4 36 theme proteoglycans 552:564 arg1 decorin 575:581 decorin 575:581 decorin 575:581 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 36 theme proteoglycans 552:564 arg1 preparations 513:524 commercially sourced preparations 492:524 commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs) 492:572 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 4 36 theme proteoglycans 552:564 arg1 biglycan 587:594 biglycan 587:594 biglycan 587:594 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 7 37 theme unexpected 1036:1045 arg1 effects 1047:1053 The unexpected effects 1032:1053 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth 1032:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 0 38 from Contaminants 0:11 arg1 preparations 27:38 commercial preparations 16:38 commercial preparations of 'purified' small leucine-rich proteoglycans 16:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 11 39 theme impurities 1767:1776 arg1 effects 1750:1756 the effects 1746:1756 the effects of these impurities 1746:1776 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 8 40 contain contains 1269:1276 arg2 sulfate 1286:1292 keratan sulfate 1278:1292 keratan sulfate 1278:1292 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 40 contain contains 1269:1276 arg1 fibromodulin 1251:1262 fibromodulin 1251:1262 fibromodulin 1251:1262 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 41 contain contain 1165:1171 arg1 Decorin 1144:1150 Decorin 1144:1150 Decorin 1144:1150 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 41 contain contain 1165:1171 arg2 chains 1236:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 41 contain contain 1165:1171 arg1 biglycan 1156:1163 biglycan 1156:1163 biglycan 1156:1163 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 1 42 theme present 124:130 arg1 study 132:136 The present study 120:136 The present study 120:136 The present study reports the perplexing results that came about because of seriously impure commercially available reagents. 27994047 4 43 theme Extensive 433:441 arg1 work 443:446 Extensive work 433:446 Extensive work 433:446 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 7 44 theme cell 1092:1095 arg1 growth 1097:1102 nerve cell growth 1086:1102 nerve cell growth 1086:1102 The unexpected effects of 'decorin' and 'biglycan' on nerve cell growth could be explained by these impurities. 27994047 9 45 theme impurities 1464:1473 arg1 structure 1411:1419 different structure 1401:1419 different structure 1401:1419 The different structure, molecular weight and composition of these impurities significantly affected our work and any conclusions that could be made. 27994047 9 45 theme impurities 1464:1473 arg1 weight 1432:1437 molecular weight 1422:1437 molecular weight 1422:1437 The different structure, molecular weight and composition of these impurities significantly affected our work and any conclusions that could be made. 27994047 9 45 theme impurities 1464:1473 arg1 composition 1443:1453 composition 1443:1453 composition 1443:1453 The different structure, molecular weight and composition of these impurities significantly affected our work and any conclusions that could be made. 27994047 11 46 dep researchers 1880:1890 arg1 time 1903:1906 time 1903:1906 time 1903:1906 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 11 46 dep researchers 1880:1890 arg1 funds 1893:1897 funds 1893:1897 funds 1893:1897 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 8 47 theme keratan 1278:1284 arg1 sulfate 1286:1292 keratan sulfate 1278:1292 keratan sulfate 1278:1292 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 8 48 theme chondroitin 1180:1190 arg1 chains 1236:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains 1173:1241 Decorin and biglycan contain either chondroitin or dermatan sulfate glycosaminoglycan (GAG) chains whereas fibromodulin only contains keratan sulfate and the large (>2500 kDa), highly glycosylated aggrecan contains both keratan and chondroitin sulfate. 27994047 10 49 theme reagent 1660:1666 arg1 purity 1623:1628 the purity 1619:1628 the purity of each commercially obtained reagent used in their experiments 1619:1692 These findings beg the question as to whether scientists need to verify the purity of each commercially obtained reagent used in their experiments. 27994047 0 50 theme leucine-rich 60:71 arg1 proteoglycans 73:85 'purified' small leucine-rich proteoglycans 43:85 'purified' small leucine-rich proteoglycans 43:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 5 51 theme various 840:846 arg1 deglycosylations 858:873 various enzymatic deglycosylations 840:873 various enzymatic deglycosylations 840:873 Unusual results compelled us to analyse the composition and purity of both preparations of these proteoglycans (PGs) using both mass spectrometry (MS) and Western blotting, with and without various enzymatic deglycosylations. 27994047 6 52 contain contain 926:932 arg1 biglycan 902:909 'biglycan' 901:910 'biglycan' 901:910 Commercial 'decorin' and 'biglycan' were found to contain a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan. 27994047 6 52 contain contain 926:932 arg1 decorin 888:894 Commercial 'decorin' 876:895 Commercial 'decorin' 876:895 Commercial 'decorin' and 'biglycan' were found to contain a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan. 27994047 6 52 contain contain 926:932 arg2 mixture 936:942 a mixture 934:942 a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan 934:1029 Commercial 'decorin' and 'biglycan' were found to contain a mixture of PGs including not only both decorin and biglycan but also fibromodulin and aggrecan. 27994047 11 53 theme researchers 1880:1890 arg1 loss 1872:1875 concomitant loss 1860:1875 concomitant loss of researchers' funds and time 1860:1906 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 11 54 theme concomitant 1860:1870 arg1 loss 1872:1875 concomitant loss 1860:1875 concomitant loss of researchers' funds and time 1860:1906 The implications of these findings are vast, since the effects of these impurities may already have led to inaccurate conclusions and reports in the literature with concomitant loss of researchers' funds and time. 27994047 4 55 theme cell 637:640 arg1 growth 642:647 nerve cell growth 631:647 nerve cell growth 631:647 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 27994047 0 56 theme small 54:58 arg1 proteoglycans 73:85 'purified' small leucine-rich proteoglycans 43:85 'purified' small leucine-rich proteoglycans 43:85 Contaminants in commercial preparations of 'purified' small leucine-rich proteoglycans may distort mechanistic studies. 27994047 1 57 theme perplexing 150:159 arg1 results 161:167 the perplexing results 146:167 the perplexing results that came about because of seriously impure commercially available reagents 146:243 The present study reports the perplexing results that came about because of seriously impure commercially available reagents. 27994047 4 58 theme nerve 631:635 arg1 growth 642:647 nerve cell growth 631:647 nerve cell growth 631:647 Extensive work was carried out within our laboratory using commercially sourced preparations of the small leucine-rich proteoglycans (SLRPs), decorin and biglycan, to investigate their influence on nerve cell growth. 26878488 4 0 theme lupane 510:515 arg1 compounds 517:525 new lupane compounds 506:525 new lupane compounds 506:525 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 4 1 theme interesting 595:605 arg1 profile 619:625 an interesting bioactivity profile 592:625 an interesting bioactivity profile against human cancer cell lines 592:657 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 5 2 theme protecting 677:686 arg1 groups 688:693 the protecting groups 673:693 the protecting groups 673:693 Influence of the protecting groups on the cytotoxicity was investigated. 26878488 0 3 from Influence 0:8 arg1 regioselectivity 46:61 regioselectivity 46:61 regioselectivity of glycosylation 46:78 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 26878488 5 4 theme groups 688:693 arg1 Influence 660:668 Influence 660:668 Influence of the protecting groups on the cytotoxicity 660:713 Influence of the protecting groups on the cytotoxicity was investigated. 26878488 6 5 theme anticancer 848:857 arg1 compounds 859:867 potential anticancer compounds 838:867 potential anticancer compounds 838:867 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 2 6 used used 299:302 arg2 series 178:183 A series 176:183 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers 176:279 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 1 7 theme lupane-type 94:104 arg1 saponins 106:113 lupane-type saponins 94:113 lupane-type saponins 94:113 Synthesis of lupane-type saponins bearing the OSW-1 saponin disaccharide unit and its isomers. 26878488 6 8 theme potential 838:846 arg1 compounds 859:867 potential anticancer compounds 838:867 potential anticancer compounds 838:867 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 4 9 theme bioactivity 607:617 arg1 profile 619:625 an interesting bioactivity profile 592:625 an interesting bioactivity profile against human cancer cell lines 592:657 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 4 10 theme Cytotoxic 484:492 arg1 activity 494:501 Cytotoxic activity 484:501 Cytotoxic activity of new lupane compounds 484:525 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 3 11 theme Unexpected 358:367 arg1 preference 369:378 Unexpected preference 358:378 Unexpected preference for 1→4-linked regioisomers 358:406 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 4 12 theme new 506:508 arg1 compounds 517:525 new lupane compounds 506:525 new lupane compounds 506:525 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 6 13 theme derivatives 823:833 arg1 synthesis 767:775 the synthesis 763:775 the synthesis of various lupane-type triterpene and saponin derivatives 763:833 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 3 14 theme sugar 460:464 arg1 rings 466:470 the sugar rings 456:470 the sugar rings 456:470 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 6 15 theme saponin 815:821 arg1 derivatives 823:833 various lupane-type triterpene and saponin derivatives 780:833 derivatives 823:833 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 2 16 theme relationships 331:343 arg1 study 351:355 the structure-activity relationships (SAR) study 308:355 the structure-activity relationships (SAR) study 308:355 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 3 17 theme conformation 440:451 arg1 inversion 423:431 an unusual inversion 412:431 an unusual inversion of the conformation of the sugar rings 412:470 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 3 17 theme conformation 440:451 arg1 preference 369:378 Unexpected preference 358:378 Unexpected preference for 1→4-linked regioisomers 358:406 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 5 18 from Influence 660:668 arg1 cytotoxicity 702:713 the cytotoxicity 698:713 the cytotoxicity 698:713 Influence of the protecting groups on the cytotoxicity was investigated. 26878488 2 19 theme structure-activity 312:329 arg1 relationships 331:343 the structure-activity relationships 308:343 the structure-activity relationships (SAR) study 308:355 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 0 20 theme hydrogen 28:35 arg1 bonds 37:41 intramolecular hydrogen bonds 13:41 intramolecular hydrogen bonds 13:41 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 26878488 6 21 theme triterpene 800:809 arg1 synthesis 767:775 the synthesis 763:775 the synthesis of various lupane-type triterpene and saponin derivatives 763:833 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 1 22 theme saponins 106:113 arg1 Synthesis 81:89 Synthesis 81:89 Synthesis of lupane-type saponins 81:113 Synthesis of lupane-type saponins bearing the OSW-1 saponin disaccharide unit and its isomers. 26878488 2 23 theme lupane-type 188:198 arg1 saponins 200:207 lupane-type saponins 188:207 lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers 188:279 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 3 24 theme rings 466:470 arg1 conformation 440:451 the conformation 436:451 the conformation of the sugar rings 436:470 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 0 25 theme bonds 37:41 arg1 Influence 0:8 Influence 0:8 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 0:79 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 26878488 3 26 theme 1→4-linked 384:393 arg1 regioisomers 395:406 1→4-linked regioisomers 384:406 1→4-linked regioisomers 384:406 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 6 27 theme lupane-type 788:798 arg1 triterpene 800:809 various lupane-type triterpene and saponin derivatives 780:833 triterpene 800:809 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 3 28 theme unusual 415:421 arg1 inversion 423:431 an unusual inversion 412:431 an unusual inversion of the conformation of the sugar rings 412:470 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 6 29 theme various 780:786 arg1 triterpene 800:809 various lupane-type triterpene and saponin derivatives 780:833 triterpene 800:809 These results open the way to the synthesis of various lupane-type triterpene and saponin derivatives as potential anticancer compounds. 26878488 1 30 theme OSW-1 127:131 arg1 unit 154:157 the OSW-1 saponin disaccharide unit 123:157 the OSW-1 saponin disaccharide unit 123:157 Synthesis of lupane-type saponins bearing the OSW-1 saponin disaccharide unit and its isomers. 26878488 2 31 theme disaccharide 223:234 arg1 unit 236:239 OSW-1 disaccharide unit 217:239 OSW-1 disaccharide unit as well as its regio- and stereoisomers 217:279 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 1 32 theme saponin 133:139 arg1 unit 154:157 the OSW-1 saponin disaccharide unit 123:157 the OSW-1 saponin disaccharide unit 123:157 Synthesis of lupane-type saponins bearing the OSW-1 saponin disaccharide unit and its isomers. 26878488 2 33 theme OSW-1 217:221 arg1 unit 236:239 OSW-1 disaccharide unit 217:239 OSW-1 disaccharide unit as well as its regio- and stereoisomers 217:279 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 3 34 link 1→4-linked 384:393 arg1 regioisomers 395:406 1→4-linked regioisomers 384:406 1→4-linked regioisomers 384:406 Unexpected preference for 1→4-linked regioisomers and an unusual inversion of the conformation of the sugar rings were noted. 26878488 1 35 theme disaccharide 141:152 arg1 unit 154:157 the OSW-1 saponin disaccharide unit 123:157 the OSW-1 saponin disaccharide unit 123:157 Synthesis of lupane-type saponins bearing the OSW-1 saponin disaccharide unit and its isomers. 26878488 4 36 theme human 635:639 arg1 lines 653:657 human cancer cell lines 635:657 human cancer cell lines 635:657 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 2 37 dep relationships 331:343 arg1 SAR 346:348 SAR 346:348 SAR 346:348 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 2 38 theme saponins 200:207 arg1 series 178:183 A series 176:183 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers 176:279 A series of lupane-type saponins bearing OSW-1 disaccharide unit as well as its regio- and stereoisomers were prepared and used for the structure-activity relationships (SAR) study. 26878488 4 39 theme cell 648:651 arg1 lines 653:657 human cancer cell lines 635:657 human cancer cell lines 635:657 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 0 40 theme glycosylation 66:78 arg1 regioselectivity 46:61 regioselectivity 46:61 regioselectivity of glycosylation 46:78 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 26878488 4 41 theme compounds 517:525 arg1 activity 494:501 Cytotoxic activity 484:501 Cytotoxic activity of new lupane compounds 484:525 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 26878488 0 42 theme intramolecular 13:26 arg1 bonds 37:41 intramolecular hydrogen bonds 13:41 intramolecular hydrogen bonds 13:41 Influence of intramolecular hydrogen bonds on regioselectivity of glycosylation. 26878488 4 43 theme cancer 641:646 arg1 lines 653:657 human cancer cell lines 635:657 human cancer cell lines 635:657 Cytotoxic activity of new lupane compounds was evaluated in vitro and revealed that some saponins exhibited an interesting bioactivity profile against human cancer cell lines. 25402728 12 0 from glycosylation 1380:1392 arg1 suppression 1338:1348 suppression 1338:1348 suppression of ANXA2 at its expression or glycosylation on NPC 1338:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 12 0 from glycosylation 1380:1392 arg1 NPC 1397:1399 NPC 1397:1399 NPC 1397:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 12 1 from expression 1366:1375 arg1 suppression 1338:1348 suppression 1338:1348 suppression of ANXA2 at its expression or glycosylation on NPC 1338:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 12 1 from expression 1366:1375 arg1 NPC 1397:1399 NPC 1397:1399 NPC 1397:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 5 2 theme NPC 630:632 arg1 cells 634:638 NPC cells 630:638 NPC cells 630:638 This research is to reveal the molecular link on NPC cells that induces the immunosuppressive responses in DCs. 25402728 10 3 theme immunosuppressive 1182:1198 arg1 production 1206:1215 the immunosuppressive IL-10 production 1178:1215 the immunosuppressive IL-10 production 1178:1215 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 25402728 8 4 theme NPC 1003:1005 arg1 growth 1007:1012 NPC growth 1003:1012 NPC growth 1003:1012 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 4 5 theme epithelial 498:507 arg1 tumor 532:536 an epithelial cell-derived malignant tumor 495:536 an epithelial cell-derived malignant tumor 495:536 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 5 theme epithelial 498:507 arg1 carcinoma 476:484 Nasopharyngeal carcinoma 461:484 Nasopharyngeal carcinoma (NPC) 461:490 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 3 6 theme C-type 350:355 arg1 molecule 305:312 DC-specific intercellular adhesion molecule 270:312 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN) 270:345 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 6 theme C-type 350:355 arg1 lectin 357:362 a C-type lectin 348:362 a C-type lectin expressed on DCs 348:379 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 8 7 theme crucial 978:984 arg1 role 986:989 a crucial role 976:989 a crucial role of ANXA2 in NPC growth 976:1012 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 7 8 from glycan 824:829 arg1 ANXA2 834:838 ANXA2 834:838 ANXA2 834:838 N-linked mannose-rich glycan on ANXA2 may mediate the interaction. 25402728 8 9 from role 986:989 arg1 growth 1007:1012 NPC growth 1003:1012 NPC growth 1003:1012 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 7 10 theme N-linked 802:809 arg1 glycan 824:829 N-linked mannose-rich glycan 802:829 N-linked mannose-rich glycan on ANXA2 802:838 N-linked mannose-rich glycan on ANXA2 may mediate the interaction. 25402728 4 11 theme Nasopharyngeal 461:474 arg1 tumor 532:536 an epithelial cell-derived malignant tumor 495:536 an epithelial cell-derived malignant tumor 495:536 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 11 theme Nasopharyngeal 461:474 arg1 NPC 487:489 NPC 487:489 NPC 487:489 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 11 theme Nasopharyngeal 461:474 arg1 carcinoma 476:484 Nasopharyngeal carcinoma 461:484 Nasopharyngeal carcinoma (NPC) 461:490 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 6 12 theme NPC 760:762 arg1 cells 764:768 NPC cells 760:768 NPC cells 760:768 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 7 13 theme mannose-rich 811:822 arg1 glycan 824:829 N-linked mannose-rich glycan 802:829 N-linked mannose-rich glycan on ANXA2 802:838 N-linked mannose-rich glycan on ANXA2 may mediate the interaction. 25402728 7 14 link N-linked 802:809 arg1 glycan 824:829 N-linked mannose-rich glycan 802:829 N-linked mannose-rich glycan on ANXA2 802:838 N-linked mannose-rich glycan on ANXA2 may mediate the interaction. 25402728 0 15 theme carcinoma 41:49 arg1 cells 51:55 nasopharygeal carcinoma cells 27:55 nasopharygeal carcinoma cells 27:55 Dendritic cells respond to nasopharygeal carcinoma cells through annexin A2-recognizing DC-SIGN. 25402728 3 16 located found 438:442 arg2 structures 414:423 certain carbohydrate structures 393:423 certain carbohydrate structures which can be found on cancer cells 393:458 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 16 located found 438:442 arg1 cells 454:458 cancer cells 447:458 cancer cells 447:458 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 2 17 theme immunosuppressive 241:257 arg1 responses 259:267 immunosuppressive responses 241:267 immunosuppressive responses 241:267 However, these cells can be tuned by tumors with immunosuppressive responses. 25402728 4 18 link cell-derived 509:520 arg1 tumor 532:536 an epithelial cell-derived malignant tumor 495:536 an epithelial cell-derived malignant tumor 495:536 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 18 link cell-derived 509:520 arg1 carcinoma 476:484 Nasopharyngeal carcinoma 461:484 Nasopharyngeal carcinoma (NPC) 461:490 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 8 19 theme NPC 942:944 arg1 xenograft 946:954 NPC xenograft 942:954 NPC xenograft 942:954 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 1 20 theme Dendritic 97:105 arg1 cells 107:111 Dendritic cells 97:111 Dendritic cells (DCs) 97:117 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 1 20 theme Dendritic 97:105 arg1 DCs 114:116 DCs 114:116 DCs 114:116 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 12 21 theme ANXA2 1353:1357 arg1 suppression 1338:1348 suppression 1338:1348 suppression of ANXA2 at its expression or glycosylation on NPC 1338:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 0 22 theme Dendritic 0:8 arg1 cells 10:14 Dendritic cells 0:14 Dendritic cells 0:14 Dendritic cells respond to nasopharygeal carcinoma cells through annexin A2-recognizing DC-SIGN. 25402728 5 23 from link 622:625 arg1 cells 634:638 NPC cells 630:638 NPC cells 630:638 This research is to reveal the molecular link on NPC cells that induces the immunosuppressive responses in DCs. 25402728 8 24 theme ANXA2 994:998 arg1 role 986:989 a crucial role 976:989 a crucial role of ANXA2 in NPC growth 976:1012 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 1 25 theme cancer 170:175 arg1 immunotherapy 177:189 cancer immunotherapy 170:189 cancer immunotherapy 170:189 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 8 26 theme ANXA2 925:929 arg1 knockdown 912:920 knockdown 912:920 knockdown of ANXA2 912:929 ANXA2 was abundantly expressed in NPC, and knockdown of ANXA2 suppressed NPC xenograft in mice, suggesting a crucial role of ANXA2 in NPC growth. 25402728 9 27 theme NPC 1032:1034 arg1 cells 1036:1040 NPC cells 1032:1040 NPC cells 1032:1040 Interaction with NPC cells caused DC-SIGN activation in DCs. 25402728 11 28 theme ANXA2 1261:1265 arg1 Blockage 1231:1238 Blockage 1231:1238 Blockage of either DC-SIGN or ANXA2 1231:1265 Blockage of either DC-SIGN or ANXA2 eliminated the production of IL-10 from DCs. 25402728 3 29 theme DC-specific 270:280 arg1 molecule 305:312 DC-specific intercellular adhesion molecule 270:312 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN) 270:345 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 29 theme DC-specific 270:280 arg1 lectin 357:362 a C-type lectin 348:362 a C-type lectin expressed on DCs 348:379 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 30 theme carbohydrate 401:412 arg1 structures 414:423 certain carbohydrate structures 393:423 certain carbohydrate structures which can be found on cancer cells 393:458 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 4 31 theme immune 548:553 arg1 response 555:562 immune response 548:562 immune response 548:562 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 0 32 theme nasopharygeal 27:39 arg1 cells 51:55 nasopharygeal carcinoma cells 27:55 nasopharygeal carcinoma cells 27:55 Dendritic cells respond to nasopharygeal carcinoma cells through annexin A2-recognizing DC-SIGN. 25402728 4 33 theme malignant 522:530 arg1 tumor 532:536 an epithelial cell-derived malignant tumor 495:536 an epithelial cell-derived malignant tumor 495:536 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 33 theme malignant 522:530 arg1 carcinoma 476:484 Nasopharyngeal carcinoma 461:484 Nasopharyngeal carcinoma (NPC) 461:490 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 6 34 from ligand 775:780 arg1 DCs 797:799 DCs 797:799 DCs 797:799 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 6 35 theme A2 746:747 arg1 ligand 775:780 a ligand 773:780 a ligand for DC-SIGN on DCs 773:799 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 6 35 theme A2 746:747 arg1 identification 720:733 identification 720:733 identification of annexin A2 (ANXA2) 720:755 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 3 36 theme cancer 447:452 arg1 cells 454:458 cancer cells 447:458 cancer cells 447:458 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 11 37 theme IL-10 1296:1300 arg1 production 1282:1291 the production 1278:1291 the production of IL-10 from DCs 1278:1309 Blockage of either DC-SIGN or ANXA2 eliminated the production of IL-10 from DCs. 25402728 4 38 theme cell-derived 509:520 arg1 tumor 532:536 an epithelial cell-derived malignant tumor 495:536 an epithelial cell-derived malignant tumor 495:536 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 4 38 theme cell-derived 509:520 arg1 carcinoma 476:484 Nasopharyngeal carcinoma 461:484 Nasopharyngeal carcinoma (NPC) 461:490 Nasopharyngeal carcinoma (NPC) is an epithelial cell-derived malignant tumor, in which immune response remains unclear. 25402728 6 39 theme annexin 738:744 arg1 ANXA2 750:754 ANXA2 750:754 ANXA2 750:754 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 6 39 theme annexin 738:744 arg1 A2 746:747 annexin A2 738:747 annexin A2 (ANXA2) 738:755 In this article, we report identification of annexin A2 (ANXA2) on NPC cells as a ligand for DC-SIGN on DCs. 25402728 11 40 theme DC-SIGN 1250:1256 arg1 Blockage 1231:1238 Blockage 1231:1238 Blockage of either DC-SIGN or ANXA2 1231:1265 Blockage of either DC-SIGN or ANXA2 eliminated the production of IL-10 from DCs. 25402728 3 41 theme certain 393:399 arg1 structures 414:423 certain carbohydrate structures 393:423 certain carbohydrate structures which can be found on cancer cells 393:458 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 5 42 theme immunosuppressive 657:673 arg1 responses 675:683 the immunosuppressive responses 653:683 the immunosuppressive responses in DCs 653:690 This research is to reveal the molecular link on NPC cells that induces the immunosuppressive responses in DCs. 25402728 11 43 from DCs 1307:1309 arg1 production 1282:1291 the production 1278:1291 the production of IL-10 from DCs 1278:1309 Blockage of either DC-SIGN or ANXA2 eliminated the production of IL-10 from DCs. 25402728 5 44 theme molecular 612:620 arg1 link 622:625 the molecular link 608:625 the molecular link on NPC cells that induces the immunosuppressive responses in DCs 608:690 This research is to reveal the molecular link on NPC cells that induces the immunosuppressive responses in DCs. 25402728 12 45 gly glycosylation 1380:1392 arg1 NPC 1397:1399 NPC 1397:1399 NPC 1397:1399 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 2 46 with tumors 229:234 arg1 responses 259:267 immunosuppressive responses 241:267 immunosuppressive responses 241:267 However, these cells can be tuned by tumors with immunosuppressive responses. 25402728 3 47 theme intercellular 282:294 arg1 molecule 305:312 DC-specific intercellular adhesion molecule 270:312 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN) 270:345 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 47 theme intercellular 282:294 arg1 lectin 357:362 a C-type lectin 348:362 a C-type lectin expressed on DCs 348:379 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 9 48 with Interaction 1015:1025 arg1 cells 1036:1040 NPC cells 1032:1040 NPC cells 1032:1040 Interaction with NPC cells caused DC-SIGN activation in DCs. 25402728 5 49 from responses 675:683 arg1 DCs 688:690 DCs 688:690 DCs 688:690 This research is to reveal the molecular link on NPC cells that induces the immunosuppressive responses in DCs. 25402728 3 50 theme adhesion 296:303 arg1 molecule 305:312 DC-specific intercellular adhesion molecule 270:312 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN) 270:345 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 3 50 theme adhesion 296:303 arg1 lectin 357:362 a C-type lectin 348:362 a C-type lectin expressed on DCs 348:379 DC-specific intercellular adhesion molecule 3-Grabbing Nonintegrin (DC-SIGN), a C-type lectin expressed on DCs, recognizes certain carbohydrate structures which can be found on cancer cells. 25402728 9 51 theme DC-SIGN 1049:1055 arg1 activation 1057:1066 DC-SIGN activation 1049:1066 DC-SIGN activation 1049:1066 Interaction with NPC cells caused DC-SIGN activation in DCs. 25402728 1 52 theme essential 127:135 arg1 role 137:140 an essential role 124:140 an essential role 124:140 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 10 53 theme proinflammatory 1111:1125 arg1 IL 1140:1141 IL 1140:1141 IL 1140:1141 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 25402728 10 53 theme proinflammatory 1111:1125 arg1 interleukin 1127:1137 the proinflammatory interleukin 1107:1137 the proinflammatory interleukin (IL)-12 production 1107:1156 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 25402728 0 54 theme A2-recognizing 73:86 arg1 DC-SIGN 88:94 annexin A2-recognizing DC-SIGN 65:94 annexin A2-recognizing DC-SIGN 65:94 Dendritic cells respond to nasopharygeal carcinoma cells through annexin A2-recognizing DC-SIGN. 25402728 12 55 theme DC-mediated 1413:1423 arg1 immunotherapy 1425:1437 DC-mediated immunotherapy 1413:1437 DC-mediated immunotherapy for the tumor 1413:1451 This report suggests that suppression of ANXA2 at its expression or glycosylation on NPC may improve DC-mediated immunotherapy for the tumor. 25402728 1 56 used used 162:165 arg2 DCs 114:116 DCs 114:116 DCs 114:116 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 1 56 used used 162:165 arg2 cells 107:111 Dendritic cells 97:111 Dendritic cells (DCs) 97:117 Dendritic cells (DCs) play an essential role in immunity and are used in cancer immunotherapy. 25402728 0 57 theme annexin 65:71 arg1 DC-SIGN 88:94 annexin A2-recognizing DC-SIGN 65:94 annexin A2-recognizing DC-SIGN 65:94 Dendritic cells respond to nasopharygeal carcinoma cells through annexin A2-recognizing DC-SIGN. 25402728 10 58 theme interleukin 1127:1137 arg1 production 1147:1156 the proinflammatory interleukin (IL)-12 production 1107:1156 the proinflammatory interleukin (IL)-12 production 1107:1156 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 25402728 10 59 theme IL-10 1200:1204 arg1 production 1206:1215 the immunosuppressive IL-10 production 1178:1215 the immunosuppressive IL-10 production 1178:1215 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 25402728 10 60 theme DC 1089:1090 arg1 maturation 1092:1101 DC maturation 1089:1101 DC maturation 1089:1101 Consequently DC maturation and the proinflammatory interleukin (IL)-12 production were inhibited, and the immunosuppressive IL-10 production was promoted. 26916835 3 0 theme glycated 465:472 arg1 Properties 438:447 Properties 438:447 Properties of the obtained glycated 438:472 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 3 0 theme glycated 465:472 arg1 GC-SPI 496:501 GC-SPI 496:501 GC-SPI 496:501 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 3 0 theme glycated 465:472 arg1 SPI 491:493 cross-linked SPI 478:493 cross-linked SPI (GC-SPI) 478:502 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 4 1 theme RESULTS 536:542 arg1 GC-SPI 544:549 RESULTS GC-SPI 536:549 RESULTS GC-SPI 536:549 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 8 2 theme protein 1418:1424 arg1 ingredient 1426:1435 potential protein ingredient 1408:1435 potential protein ingredient with good hydration and dispersive stabilisation 1408:1484 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 5 3 contain had 772:774 arg1 GC-SPI 729:734 GC-SPI 729:734 GC-SPI other than SPI and cross-linked SPI 729:770 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 5 3 contain had 772:774 arg1 SPI 747:749 SPI 747:749 SPI 747:749 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 5 3 contain had 772:774 arg2 OH 787:788 more &bond;OH 776:788 more &bond;OH in molecules 776:801 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 5 4 theme Infrared 669:676 arg1 spectra 678:684 Infrared spectra 669:684 Infrared spectra 669:684 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 8 5 theme potential 1408:1416 arg1 ingredient 1426:1435 potential protein ingredient 1408:1435 potential protein ingredient with good hydration and dispersive stabilisation 1408:1484 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 3 6 theme cross-linked 478:489 arg1 GC-SPI 496:501 GC-SPI 496:501 GC-SPI 496:501 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 3 6 theme cross-linked 478:489 arg1 SPI 491:493 cross-linked SPI 478:493 cross-linked SPI (GC-SPI) 478:502 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 6 7 theme water-binding 899:911 arg1 capacity 913:920 enhanced water-binding capacity 890:920 enhanced water-binding capacity 890:920 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 8 dep radius 972:977 arg1 180.2 980:984 180.2 980:984 180.2 980:984 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 8 dep radius 972:977 arg1 82.9 nm 990:996 82.9 nm 990:996 82.9 nm 990:996 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 9 theme lower 1077:1081 arg1 stability 1091:1099 lower thermal stability 1077:1099 lower thermal stability (e.g. greater mass loss) 1077:1124 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 10 theme protein 1247:1253 arg1 cross-linking 1255:1267 the protein cross-linking 1243:1267 the protein cross-linking 1243:1267 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 11 dep greater 1107:1113 arg1 e.g. 1102:1105 e.g. 1102:1105 e.g. 1102:1105 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 11 dep greater 1107:1113 arg1 loss 1120:1123 mass loss 1115:1123 mass loss 1115:1123 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 12 theme 288 °C. 1162:1168 arg1 GC-SPI 1170:1175 288 °C. GC-SPI 1162:1175 288 °C. GC-SPI 1162:1175 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 5 13 theme &bond 781:785 arg1 OH 787:788 more &bond;OH 776:788 more &bond;OH in molecules 776:801 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 6 14 theme thermal 1083:1089 arg1 stability 1091:1099 lower thermal stability 1077:1099 lower thermal stability (e.g. greater mass loss) 1077:1124 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 4 15 theme than 626:629 arg1 protein 659:665 0.42 vs. 0.50 mol kg-1 protein 636:665 0.42 vs. 0.50 mol kg-1 protein 636:665 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 4 15 theme than 626:629 arg1 SPI 631:633 NH2 than SPI 622:633 NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein) 622:666 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 1 16 theme important 151:159 arg1 ingredient 169:178 an important protein ingredient 148:178 an important protein ingredient for the food industry 148:200 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 1 16 theme important 151:159 arg1 protein 137:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 0 17 theme cross-linked 38:49 arg1 protein 55:61 a glycated and cross-linked soy protein 23:61 a glycated and cross-linked soy protein 23:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 6 18 from temperature 1144:1154 arg1 heating 1131:1137 heating 1131:1137 heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking 1131:1267 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 5 19 theme cross-linked 755:766 arg1 SPI 768:770 cross-linked SPI 755:770 cross-linked SPI 755:770 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 1 20 theme protein 161:167 arg1 ingredient 169:178 an important protein ingredient 148:178 an important protein ingredient for the food industry 148:200 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 1 20 theme protein 161:167 arg1 protein 137:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 8 21 theme good 1442:1445 arg1 hydration 1447:1455 good hydration 1442:1455 good hydration 1442:1455 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 6 22 theme negative 1003:1010 arg1 mV 1044:1045 -31.2 vs. -27.7 mV 1028:1045 -31.2 vs. -27.7 mV 1028:1045 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 6 22 theme negative 1003:1010 arg1 zeta-potential 1012:1025 negative zeta-potential 1003:1025 negative zeta-potential (-31.2 vs. -27.7 mV) 1003:1046 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 0 23 link cross-linked 38:49 arg1 protein 55:61 a glycated and cross-linked soy protein 23:61 a glycated and cross-linked soy protein 23:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 7 24 theme transglutaminase 1308:1323 arg1 Oligochitosan 1281:1293 CONCLUSION Oligochitosan 1270:1293 CONCLUSION Oligochitosan of 5 kDa and transglutaminase 1270:1323 CONCLUSION Oligochitosan of 5 kDa and transglutaminase can be used to glycate and cross-link SPI. 26916835 2 25 theme protein 361:367 arg1 isolate 369:375 soy protein isolate 357:375 soy protein isolate (SPI) 357:381 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 2 25 theme protein 361:367 arg1 SPI 378:380 SPI 378:380 SPI 378:380 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 0 26 theme Modified 0:7 arg1 properties 9:18 Modified properties 0:18 Modified properties of a glycated and cross-linked soy protein 0:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 1 27 theme enzymatic 246:254 arg1 modifications 269:281 enzymatic and chemical modifications 246:281 enzymatic and chemical modifications 246:281 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 5 28 dep other 736:740 arg1 than 742:745 than 742:745 than 742:745 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 6 29 with comparison 855:864 arg1 SPI 871:873 SPI 871:873 SPI 871:873 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 2 30 theme soy 357:359 arg1 isolate 369:375 soy protein isolate 357:375 soy protein isolate (SPI) 357:381 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 2 30 theme soy 357:359 arg1 SPI 378:380 SPI 378:380 SPI 378:380 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 6 31 theme enhanced 890:897 arg1 capacity 913:920 enhanced water-binding capacity 890:920 enhanced water-binding capacity 890:920 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 0 32 theme kDa 117:119 arg1 transglutaminase 74:89 transglutaminase 74:89 transglutaminase 74:89 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 0 32 theme kDa 117:119 arg1 oligochitosan 98:110 an oligochitosan 95:110 an oligochitosan of 5 kDa 95:119 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 7 33 theme kDa 1300:1302 arg1 Oligochitosan 1281:1293 CONCLUSION Oligochitosan 1270:1293 CONCLUSION Oligochitosan of 5 kDa and transglutaminase 1270:1323 CONCLUSION Oligochitosan of 5 kDa and transglutaminase can be used to glycate and cross-link SPI. 26916835 8 34 with ingredient 1426:1435 arg1 stabilisation 1472:1484 dispersive stabilisation 1461:1484 dispersive stabilisation 1461:1484 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 8 34 with ingredient 1426:1435 arg1 hydration 1447:1455 good hydration 1442:1455 good hydration 1442:1455 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 1 35 theme chemical 260:267 arg1 modifications 269:281 enzymatic and chemical modifications 246:281 enzymatic and chemical modifications 246:281 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 6 36 theme due 1236:1238 arg1 SPI 1232:1234 SPI 1232:1234 SPI due to the protein cross-linking 1232:1267 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 4 37 dep contained 551:559 arg1 protein 659:665 0.42 vs. 0.50 mol kg-1 protein 636:665 0.42 vs. 0.50 mol kg-1 protein 636:665 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 4 37 dep contained 551:559 arg1 SPI 631:633 NH2 than SPI 622:633 NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein) 622:666 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 1 38 theme food 188:191 arg1 industry 193:200 the food industry 184:200 the food industry 184:200 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 3 39 theme obtained 456:463 arg1 glycated 465:472 the obtained glycated 452:472 the obtained glycated 452:472 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 3 40 link cross-linked 478:489 arg1 GC-SPI 496:501 GC-SPI 496:501 GC-SPI 496:501 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 3 40 link cross-linked 478:489 arg1 SPI 491:493 cross-linked SPI 478:493 cross-linked SPI (GC-SPI) 478:502 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 9 41 theme Chemical 1505:1512 arg1 Industry 1514:1521 Chemical Industry 1505:1521 Chemical Industry 1505:1521 © 2016 Society of Chemical Industry. 26916835 4 42 theme NH2 622:624 arg1 protein 659:665 0.42 vs. 0.50 mol kg-1 protein 636:665 0.42 vs. 0.50 mol kg-1 protein 636:665 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 4 42 theme NH2 622:624 arg1 SPI 631:633 NH2 than SPI 622:633 NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein) 622:666 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 4 43 theme reactable 606:614 arg1 &bond 616:620 less reactable &bond 601:620 less reactable &bond 601:620 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 2 44 theme transglutaminase 420:435 arg1 oligochitosan 393:405 an oligochitosan 390:405 an oligochitosan of 5 kDa and transglutaminase 390:435 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 0 45 theme glycated 25:32 arg1 protein 55:61 a glycated and cross-linked soy protein 23:61 a glycated and cross-linked soy protein 23:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 4 46 contain contained 551:559 arg1 GC-SPI 544:549 RESULTS GC-SPI 536:549 RESULTS GC-SPI 536:549 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 4 46 contain contained 551:559 arg2 glucosamine 561:571 glucosamine 561:571 glucosamine of 13.6 g kg-1 protein 561:594 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 5 47 theme circular 690:697 arg1 dichroism 699:707 circular dichroism 690:707 circular dichroism 690:707 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 0 48 theme soy 51:53 arg1 protein 55:61 a glycated and cross-linked soy protein 23:61 a glycated and cross-linked soy protein 23:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 7 49 theme CONCLUSION 1270:1279 arg1 Oligochitosan 1281:1293 CONCLUSION Oligochitosan 1270:1293 CONCLUSION Oligochitosan of 5 kDa and transglutaminase 1270:1323 CONCLUSION Oligochitosan of 5 kDa and transglutaminase can be used to glycate and cross-link SPI. 26916835 5 50 from OH 787:788 arg1 molecules 793:801 molecules 793:801 molecules 793:801 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 5 51 theme secondary 831:839 arg1 structure 841:849 secondary structure 831:849 secondary structure 831:849 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 2 52 theme kDa 412:414 arg1 oligochitosan 393:405 an oligochitosan 390:405 an oligochitosan of 5 kDa and transglutaminase 390:435 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 6 53 theme proteolytic 1201:1211 arg1 digestibility 1213:1225 vitro proteolytic digestibility 1195:1225 vitro proteolytic digestibility 1195:1225 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 2 54 theme enzymatic 309:317 arg1 glycation 319:327 a new enzymatic glycation 303:327 a new enzymatic glycation 303:327 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 6 55 theme vitro 1195:1199 arg1 digestibility 1213:1225 vitro proteolytic digestibility 1195:1225 vitro proteolytic digestibility 1195:1225 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 8 56 theme dispersive 1461:1470 arg1 stabilisation 1472:1484 dispersive stabilisation 1461:1484 dispersive stabilisation 1461:1484 This approach is applicable to generate potential protein ingredient with good hydration and dispersive stabilisation. 26916835 2 57 theme new 305:307 arg1 glycation 319:327 a new enzymatic glycation 303:327 a new enzymatic glycation 303:327 This study applied a new enzymatic glycation and cross-linking to modify soy protein isolate (SPI), using an oligochitosan of 5 kDa and transglutaminase. 26916835 6 58 theme hydrodynamic 959:970 arg1 radius 972:977 enlarged hydrodynamic radius 950:977 enlarged hydrodynamic radius (180.2 vs. 82.9 nm) 950:997 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 5 59 dep spectra 678:684 arg1 results 709:715 results 709:715 results 709:715 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 6 60 theme mass 1115:1118 arg1 loss 1120:1123 mass loss 1115:1123 mass loss 1115:1123 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 1 61 theme BACKGROUND 122:131 arg1 ingredient 169:178 an important protein ingredient 148:178 an important protein ingredient for the food industry 148:200 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 1 61 theme BACKGROUND 122:131 arg1 protein 137:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 6 62 theme enlarged 950:957 arg1 radius 972:977 enlarged hydrodynamic radius 950:977 enlarged hydrodynamic radius (180.2 vs. 82.9 nm) 950:997 In comparison with SPI, GC-SPI showed enhanced water-binding capacity, could form aggregates with enlarged hydrodynamic radius (180.2 vs. 82.9 nm) and negative zeta-potential (-31.2 vs. -27.7 mV) in dispersion, but exhibited lower thermal stability (e.g. greater mass loss) upon heating at a temperature above 288 °C. GC-SPI also had lower in vitro proteolytic digestibility than SPI due to the protein cross-linking. 26916835 7 63 used used 1332:1335 arg2 Oligochitosan 1281:1293 CONCLUSION Oligochitosan 1270:1293 CONCLUSION Oligochitosan of 5 kDa and transglutaminase 1270:1323 CONCLUSION Oligochitosan of 5 kDa and transglutaminase can be used to glycate and cross-link SPI. 26916835 3 64 dep unknown 509:515 arg1 assessed 526:533 assessed 526:533 assessed 526:533 Properties of the obtained glycated and cross-linked SPI (GC-SPI) were unknown and thus assessed. 26916835 1 65 theme Soy 133:135 arg1 ingredient 169:178 an important protein ingredient 148:178 an important protein ingredient for the food industry 148:200 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 1 65 theme Soy 133:135 arg1 protein 137:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein 122:143 BACKGROUND Soy protein is an important protein ingredient for the food industry; however, its properties can be improved by enzymatic and chemical modifications. 26916835 4 66 theme protein 588:594 arg1 glucosamine 561:571 glucosamine 561:571 glucosamine of 13.6 g kg-1 protein 561:594 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 26916835 0 67 theme protein 55:61 arg1 properties 9:18 Modified properties 0:18 Modified properties of a glycated and cross-linked soy protein 0:61 Modified properties of a glycated and cross-linked soy protein isolate by transglutaminase and an oligochitosan of 5 kDa. 26916835 5 68 link cross-linked 755:766 arg1 SPI 768:770 cross-linked SPI 755:770 cross-linked SPI 755:770 Infrared spectra and circular dichroism results showed that GC-SPI other than SPI and cross-linked SPI had more &bond;OH in molecules, and was more disordered in secondary structure. 26916835 4 69 theme 13.6 g kg-1 576:586 arg1 protein 588:594 13.6 g kg-1 protein 576:594 13.6 g kg-1 protein 576:594 RESULTS GC-SPI contained glucosamine of 13.6 g kg-1 protein, but less reactable &bond;NH2 than SPI (0.42 vs. 0.50 mol kg-1 protein). 24721075 3 0 link -linked 483:489 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 0 link -linked 483:489 arg1 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 1 1 from teas 185:188 arg1 folk 207:210 the folk 203:210 the folk of China for centuries 203:233 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 6 2 theme sulfated 948:955 arg1 derivatives 957:967 the sulfated derivatives 944:967 the sulfated derivatives 944:967 Moreover, the sulfated derivatives exhibited stronger inhibitory abilities on protein glycation than those of DHPD2. 24721075 0 3 theme polysaccharide 95:108 arg1 identification 11:24 Structural identification 0:24 Structural identification 0:24 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 0 3 theme polysaccharide 95:108 arg1 modification 39:50 sulfated modification 30:50 sulfated modification 30:50 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 3 4 theme -linked 557:563 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 4 theme -linked 557:563 arg1 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 5 dep β-d-Glcp 491:498 arg1 1→6 479:481 1→6 479:481 1→6 479:481 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 6 link -linked 531:537 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 6 link -linked 531:537 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 2 7 with DHPD2 290:294 arg1 weight 311:316 molecular weight 301:316 molecular weight of 8.09 × 10(6)Da 301:334 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 3 8 link -linked 460:466 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 8 link -linked 460:466 arg1 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 5 9 from appearance 783:792 arg1 -1 860:861 -1 860:861 -1 860:861 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 5 9 from appearance 783:792 arg1 822cm 854:858 822cm 854:858 822cm(-1) 854:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 5 9 from appearance 783:792 arg1 1250 845:848 near 1250 840:848 near 1250 840:848 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 5 9 from appearance 783:792 arg1 spectra 873:879 FT-IR spectra 867:879 FT-IR spectra 867:879 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 2 10 theme polysaccharide 275:288 arg1 DHPD2 290:294 an antiglycation polysaccharide DHPD2 258:294 an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da 258:334 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 6 11 theme protein 1012:1018 arg1 glycation 1020:1028 protein glycation 1012:1028 protein glycation 1012:1028 Moreover, the sulfated derivatives exhibited stronger inhibitory abilities on protein glycation than those of DHPD2. 24721075 3 12 theme -linked 483:489 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 12 theme -linked 483:489 arg1 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 1 13 used used 172:175 arg2 huoshanense 122:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 1 13 used used 172:175 arg2 material 153:160 an important food material 135:160 an important food material 135:160 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 2 14 theme antiglycation 261:273 arg1 DHPD2 290:294 an antiglycation polysaccharide DHPD2 258:294 an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da 258:334 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 6 15 theme stronger 979:986 arg1 abilities 999:1007 stronger inhibitory abilities 979:1007 stronger inhibitory abilities 979:1007 Moreover, the sulfated derivatives exhibited stronger inhibitory abilities on protein glycation than those of DHPD2. 24721075 3 16 theme terminal 596:603 arg1 α-d-Xlyp 605:612 terminal α-d-Xlyp 596:612 terminal α-d-Xlyp 596:612 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 17 theme -linked 460:466 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 17 theme -linked 460:466 arg1 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 6 18 theme inhibitory 988:997 arg1 abilities 999:1007 stronger inhibitory abilities 979:1007 stronger inhibitory abilities 979:1007 Moreover, the sulfated derivatives exhibited stronger inhibitory abilities on protein glycation than those of DHPD2. 24721075 5 19 theme absorption 820:829 arg1 bands 831:835 two new characteristic absorption bands 797:835 two new characteristic absorption bands at near 1250 and 822cm(-1) 797:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 3 20 theme -linked 437:443 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 20 theme -linked 437:443 arg1 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 20 theme -linked 437:443 arg1 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 20 theme -linked 437:443 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 20 theme -linked 437:443 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 20 theme -linked 437:443 arg1 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 7 21 theme NMR 1051:1053 arg1 analysis 1055:1062 NMR analysis 1051:1062 NMR analysis 1051:1062 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 3 22 link -linked 506:512 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 22 link -linked 506:512 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 4 23 theme substitution 739:750 arg1 degree 752:757 the substitution degree 735:757 the substitution degree of 0.475 and 0.940 735:776 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 1 24 theme Dendrobium 111:120 arg1 material 153:160 an important food material 135:160 an important food material 135:160 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 1 24 theme Dendrobium 111:120 arg1 huoshanense 122:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 0 25 theme Structural 0:9 arg1 identification 11:24 Structural identification 0:24 Structural identification 0:24 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 3 26 theme β-d-Manp 618:625 arg1 branches 584:591 the branches 580:591 the branches of terminal α-d-Xlyp and β-d-Manp 580:625 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 2 27 theme molecular 301:309 arg1 weight 311:316 molecular weight 301:316 molecular weight of 8.09 × 10(6)Da 301:334 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 3 28 link -linked 557:563 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 28 link -linked 557:563 arg1 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 0 29 theme sulfated 30:37 arg1 modification 39:50 sulfated modification 30:50 sulfated modification 30:50 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 3 30 dep β-d-Galp 565:572 arg1 1→6 553:555 1→6 553:555 1→6 553:555 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 2 31 dep 10 328:329 arg1 6 331:331 6 331:331 6 331:331 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 2 32 theme D. 384:385 arg1 huoshanense 387:397 D. huoshanense 384:397 D. huoshanense 384:397 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 2 33 theme huoshanense 387:397 arg1 bodies 374:379 the protocorm-like bodies 355:379 the protocorm-like bodies of D. huoshanense 355:397 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 3 34 theme α-d-Xlyp 605:612 arg1 branches 584:591 the branches 580:591 the branches of terminal α-d-Xlyp and β-d-Manp 580:625 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 5 35 theme bands 831:835 arg1 appearance 783:792 The appearance 779:792 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra 779:879 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 3 36 theme -linked 531:537 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 36 theme -linked 531:537 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 5 37 theme near 840:843 arg1 1250 845:848 near 1250 840:848 near 1250 840:848 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 7 38 theme sugar 1109:1113 arg1 residues 1115:1122 sugar residues 1109:1122 sugar residues 1109:1122 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 3 39 dep α-d-Glcp 468:475 arg1 1→6 456:458 1→6 456:458 1→6 456:458 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 5 40 from 1250 845:848 arg1 appearance 783:792 The appearance 779:792 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra 779:879 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 5 40 from 1250 845:848 arg1 bands 831:835 two new characteristic absorption bands 797:835 two new characteristic absorption bands at near 1250 and 822cm(-1) 797:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 3 41 theme 1→4 502:504 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 41 theme 1→4 502:504 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 42 theme 1→3,6 525:529 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 42 theme 1→3,6 525:529 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 2 43 theme protocorm-like 359:372 arg1 bodies 374:379 the protocorm-like bodies 355:379 the protocorm-like bodies of D. huoshanense 355:397 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 0 44 theme antiglycation 58:70 arg1 polysaccharide 95:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 3 45 contain contained 422:430 arg2 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg2 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg1 backbone 404:411 The backbone 400:411 The backbone of DHPD2 400:420 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg2 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg2 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg2 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 45 contain contained 422:430 arg2 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 4 46 theme sulfated 709:716 arg1 derivatives 718:728 two sulfated derivatives 705:728 two sulfated derivatives 705:728 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 1 47 theme important 138:146 arg1 material 153:160 an important food material 135:160 an important food material 135:160 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 1 47 theme important 138:146 arg1 huoshanense 122:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 4 48 theme 0.940 772:776 arg1 degree 752:757 the substitution degree 735:757 the substitution degree of 0.475 and 0.940 735:776 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 7 49 from sulfation 1083:1091 arg1 C6 1103:1104 C6 1103:1104 C6 1103:1104 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 7 49 from sulfation 1083:1091 arg1 C2 1096:1097 C2 1096:1097 C2 1096:1097 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 1 50 theme food 148:151 arg1 material 153:160 an important food material 135:160 an important food material 135:160 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 1 50 theme food 148:151 arg1 huoshanense 122:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense 111:132 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 5 51 theme FT-IR 867:871 arg1 spectra 873:879 FT-IR spectra 867:879 FT-IR spectra 867:879 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 3 52 link -linked 437:443 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 52 link -linked 437:443 arg1 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 52 link -linked 437:443 arg1 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 52 link -linked 437:443 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 52 link -linked 437:443 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 52 link -linked 437:443 arg1 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 5 53 theme sulfation 905:913 arg1 success 894:900 the success 890:900 the success of sulfation occurred to DHPD2 890:931 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 4 54 theme chlorosulfonic 661:674 arg1 method 690:695 chlorosulfonic acid-pyridine method 661:695 chlorosulfonic acid-pyridine method 661:695 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 3 55 theme 1→5 433:435 arg1 β-d-Galp 539:546 (1→3,6)-linked β-d-Galp 524:546 (1→3,6)-linked β-d-Galp 524:546 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 55 theme 1→5 433:435 arg1 β-d-Galp 565:572 (1→6)-linked β-d-Galp 552:572 (1→6)-linked β-d-Galp 552:572 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 55 theme 1→5 433:435 arg1 β-d-Glcp 491:498 (1→6)-linked β-d-Glcp 478:498 (1→6)-linked β-d-Glcp 478:498 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 55 theme 1→5 433:435 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 55 theme 1→5 433:435 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 55 theme 1→5 433:435 arg1 α-d-Glcp 468:475 (1→6)-linked α-d-Glcp 455:475 (1→6)-linked α-d-Glcp 455:475 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 1 56 theme China 215:219 arg1 folk 207:210 the folk 203:210 the folk of China for centuries 203:233 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 2 57 theme present 243:249 arg1 study 251:255 the present study 239:255 the present study 239:255 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 5 58 theme characteristic 805:818 arg1 bands 831:835 two new characteristic absorption bands 797:835 two new characteristic absorption bands at near 1250 and 822cm(-1) 797:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 4 59 theme 0.475 762:766 arg1 degree 752:757 the substitution degree 735:757 the substitution degree of 0.475 and 0.940 735:776 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 5 60 theme new 801:803 arg1 bands 831:835 two new characteristic absorption bands 797:835 two new characteristic absorption bands at near 1250 and 822cm(-1) 797:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 0 61 theme huoshanense 83:93 arg1 polysaccharide 95:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 3 62 theme DHPD2 416:420 arg1 backbone 404:411 The backbone 400:411 The backbone of DHPD2 400:420 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 63 theme -linked 506:512 arg1 β-d-Glcp 514:521 (1→4)-linked β-d-Glcp 501:521 (1→4)-linked β-d-Glcp 501:521 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 3 63 theme -linked 506:512 arg1 α-l-Araf 445:452 (1→5)-linked α-l-Araf 432:452 (1→5)-linked α-l-Araf 432:452 The backbone of DHPD2 contained (1→5)-linked α-l-Araf, (1→6)-linked α-d-Glcp, (1→6)-linked β-d-Glcp, (1→4)-linked β-d-Glcp, (1→3,6)-linked β-d-Galp and (1→6)-linked β-d-Galp, with the branches of terminal α-d-Xlyp and β-d-Manp. 24721075 1 64 from soups 194:198 arg1 folk 207:210 the folk 203:210 the folk of China for centuries 203:233 Dendrobium huoshanense, an important food material, has been used to make teas and soups in the folk of China for centuries. 24721075 2 65 theme Da 333:334 arg1 weight 311:316 molecular weight 301:316 molecular weight of 8.09 × 10(6)Da 301:334 In the present study, an antiglycation polysaccharide DHPD2 with molecular weight of 8.09 × 10(6)Da was extracted from the protocorm-like bodies of D. huoshanense. 24721075 0 66 theme Dendrobium 72:81 arg1 polysaccharide 95:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 an antiglycation Dendrobium huoshanense polysaccharide 55:108 Structural identification and sulfated modification of an antiglycation Dendrobium huoshanense polysaccharide. 24721075 4 67 theme acid-pyridine 676:688 arg1 method 690:695 chlorosulfonic acid-pyridine method 661:695 chlorosulfonic acid-pyridine method 661:695 DHPD2 was further modified using chlorosulfonic acid-pyridine method, giving two sulfated derivatives with the substitution degree of 0.475 and 0.940. 24721075 7 68 theme residues 1115:1122 arg1 C6 1103:1104 C6 1103:1104 C6 1103:1104 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 7 68 theme residues 1115:1122 arg1 C2 1096:1097 C2 1096:1097 C2 1096:1097 NMR analysis disclosed that the sulfation on C2 and C6 of sugar residues was beneficial to enhance this activity. 24721075 5 69 from 822cm 854:858 arg1 appearance 783:792 The appearance 779:792 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra 779:879 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 24721075 5 69 from 822cm 854:858 arg1 bands 831:835 two new characteristic absorption bands 797:835 two new characteristic absorption bands at near 1250 and 822cm(-1) 797:862 The appearance of two new characteristic absorption bands at near 1250 and 822cm(-1) in FT-IR spectra revealed the success of sulfation occurred to DHPD2. 26047718 5 0 theme expert 752:757 arg1 annotations 759:769 Cartoonist and expert annotations 737:769 Cartoonist and expert annotations 737:769 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 2 1 theme detail 452:457 arg1 level 443:447 the "cartoon" level 429:447 the "cartoon" level of detail 429:457 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 2 from pathways 334:341 arg1 system 361:366 the biological system 346:366 the biological system 346:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 3 theme biological 350:359 arg1 system 361:366 the biological system 346:366 the biological system 346:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 1 4 with N-glycans 139:147 arg1 degree 159:164 some degree 154:164 some degree of accuracy 154:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 3 5 theme developed 485:493 arg1 Cartoonist 460:469 Cartoonist 460:469 Cartoonist 460:469 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 3 5 theme developed 485:493 arg1 tool 523:526 a recently developed, fully automatic annotation tool 474:526 a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles 474:556 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 3 6 theme annotation 512:521 arg1 Cartoonist 460:469 Cartoonist 460:469 Cartoonist 460:469 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 3 6 theme annotation 512:521 arg1 tool 523:526 a recently developed, fully automatic annotation tool 474:526 a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles 474:556 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 2 7 from glycans 309:315 arg1 system 361:366 the biological system 346:366 the biological system 346:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 7 8 theme Proteomics 1135:1144 arg1 part 1087:1090 part 1087:1090 part of a Special Issue entitled: Computational Proteomics 1087:1144 This article is part of a Special Issue entitled: Computational Proteomics. 26047718 2 9 theme biosynthetic 321:332 arg1 pathways 334:341 biosynthetic pathways 321:341 biosynthetic pathways 321:341 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 5 10 theme fewer 846:850 arg1 structures 862:871 meaning fewer suggested structures 838:871 meaning fewer suggested structures per peak 838:880 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 1 11 theme Human 72:76 arg1 experts 78:84 Human experts 72:84 Human experts 72:84 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 5 12 theme suggested 852:860 arg1 structures 862:871 meaning fewer suggested structures 838:871 meaning fewer suggested structures per peak 838:880 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 4 13 theme human 648:652 arg1 data 673:676 human and mouse N-glycan data 648:676 human and mouse N-glycan data from the Consortium for Functional Glycomics 648:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 0 14 theme MALDI-TOF 43:51 arg1 profiles 62:69 MALDI-TOF N-glycan profiles 43:69 MALDI-TOF N-glycan profiles 43:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 2 15 from knowledge 280:288 arg1 system 361:366 the biological system 346:366 the biological system 346:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 16 from system 361:366 arg1 knowledge 280:288 expert knowledge 273:288 expert knowledge of the most common glycans and biosynthetic pathways in the biological system 273:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 17 theme expert 273:278 arg1 knowledge 280:288 expert knowledge 273:288 expert knowledge of the most common glycans and biosynthetic pathways in the biological system 273:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 1 18 theme accuracy 169:176 arg1 degree 159:164 some degree 154:164 some degree of accuracy 154:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 7 19 theme entitled 1111:1118 arg1 Proteomics 1135:1144 a Special Issue entitled: Computational Proteomics 1095:1144 a Special Issue entitled: Computational Proteomics 1095:1144 This article is part of a Special Issue entitled: Computational Proteomics. 26047718 5 20 theme Cartoonist 737:746 arg1 annotations 759:769 Cartoonist and expert annotations 737:769 Cartoonist and expert annotations 737:769 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 3 21 theme glycan 542:547 arg1 profiles 549:556 MALDI-TOF glycan profiles 532:556 MALDI-TOF glycan profiles 532:556 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 4 22 theme mouse 658:662 arg1 data 673:676 human and mouse N-glycan data 648:676 human and mouse N-glycan data from the Consortium for Functional Glycomics 648:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 0 23 theme Benchmark 0:8 arg1 study 10:14 Benchmark study 0:14 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles 0:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 3 24 theme MALDI-TOF 532:540 arg1 profiles 549:556 MALDI-TOF glycan profiles 532:556 MALDI-TOF glycan profiles 532:556 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 4 25 theme Functional 702:711 arg1 Glycomics 713:721 Functional Glycomics 702:721 Functional Glycomics 702:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 2 26 theme glycans 309:315 arg1 knowledge 280:288 expert knowledge 273:288 expert knowledge of the most common glycans and biosynthetic pathways in the biological system 273:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 0 27 theme automatic 19:27 arg1 annotation 29:38 automatic annotation 19:38 automatic annotation of MALDI-TOF N-glycan profiles 19:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 2 28 theme common 302:307 arg1 glycans 309:315 most common glycans 297:315 most common glycans 297:315 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 29 theme glycan 408:413 arg1 structures 415:424 most likely glycan structures 396:424 most likely glycan structures 396:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 30 theme structures 415:424 arg1 structures 415:424 most likely glycan structures 396:424 most likely glycan structures 396:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 30 theme structures 415:424 arg1 set 389:391 a small set 381:391 a small set of most likely glycan structures 381:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 5 31 theme annotated 933:941 arg1 peaks 943:947 Cartoonist more comprehensively, meaning more annotated peaks 887:947 Cartoonist more comprehensively, meaning more annotated peaks 887:947 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 2 32 dep glycans 309:315 arg1 the 293:295 the 293:295 the 293:295 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 33 theme likely 401:406 arg1 structures 415:424 most likely glycan structures 396:424 most likely glycan structures 396:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 4 34 from Consortium 687:696 arg1 data 673:676 human and mouse N-glycan data 648:676 human and mouse N-glycan data from the Consortium for Functional Glycomics 648:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 0 35 theme annotation 29:38 arg1 study 10:14 Benchmark study 0:14 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles 0:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 4 36 theme expert 626:631 arg1 annotations 633:643 human expert annotations 620:643 human expert annotations 620:643 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 2 37 theme " 441:441 arg1 level 443:447 the "cartoon" level 429:447 the "cartoon" level of detail 429:457 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 0 38 theme profiles 62:69 arg1 annotation 29:38 automatic annotation 19:38 automatic annotation of MALDI-TOF N-glycan profiles 19:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 2 39 theme intact 220:225 arg1 masses 227:232 only intact masses 215:232 only intact masses without any fragmentation information 215:270 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 1 40 theme MALDI-TOF 108:116 arg1 profiles 118:125 MALDI-TOF profiles 108:125 MALDI-TOF profiles of detached N-glycans with some degree of accuracy 108:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 2 41 theme cartoon 434:440 arg1 level 443:447 the "cartoon" level 429:447 the "cartoon" level of detail 429:457 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 42 theme only 215:218 arg1 masses 227:232 only intact masses 215:232 only intact masses without any fragmentation information 215:270 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 7 43 dep entitled 1111:1118 arg1 Computational 1121:1133 Computational 1121:1133 Computational 1121:1133 This article is part of a Special Issue entitled: Computational Proteomics. 26047718 1 44 from peaks 99:103 arg1 profiles 118:125 MALDI-TOF profiles 108:125 MALDI-TOF profiles of detached N-glycans with some degree of accuracy 108:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 2 45 theme pathways 334:341 arg1 knowledge 280:288 expert knowledge 273:288 expert knowledge of the most common glycans and biosynthetic pathways in the biological system 273:366 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 3 46 dep developed 485:493 arg1 automatic 502:510 automatic 502:510 automatic 502:510 Cartoonist is a recently developed, fully automatic annotation tool for MALDI-TOF glycan profiles. 26047718 6 47 theme unique 1005:1010 arg1 cartoons 1012:1019 unique cartoons 1005:1019 unique cartoons 1005:1019 On peaks for which both Cartoonist and the expert give unique cartoons, the two cartoons agree in over 90% of all cases. 26047718 2 48 theme MALDI-TOF 191:199 arg1 profiles 201:208 MALDI-TOF profiles 191:208 MALDI-TOF profiles 191:208 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 5 49 dep annotated 933:941 arg1 meaning 920:926 meaning 920:926 meaning 920:926 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 5 49 dep annotated 933:941 arg1 Cartoonist 887:896 Cartoonist 887:896 Cartoonist 887:896 We find that Cartoonist and expert annotations largely agree, but the expert tends to annotate more specifically, meaning fewer suggested structures per peak, and Cartoonist more comprehensively, meaning more annotated peaks. 26047718 1 50 theme detached 130:137 arg1 N-glycans 139:147 detached N-glycans 130:147 detached N-glycans with some degree of accuracy 130:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 4 51 from annotations 600:610 arg1 data 673:676 human and mouse N-glycan data 648:676 human and mouse N-glycan data from the Consortium for Functional Glycomics 648:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 2 52 theme fragmentation 246:258 arg1 information 260:270 any fragmentation information 242:270 any fragmentation information 242:270 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 4 53 theme human 620:624 arg1 annotations 633:643 human expert annotations 620:643 human expert annotations 620:643 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 1 54 theme N-glycans 139:147 arg1 profiles 118:125 MALDI-TOF profiles 108:125 MALDI-TOF profiles of detached N-glycans with some degree of accuracy 108:176 Human experts can annotate peaks in MALDI-TOF profiles of detached N-glycans with some degree of accuracy. 26047718 2 55 theme small 383:387 arg1 structures 415:424 most likely glycan structures 396:424 most likely glycan structures 396:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 2 55 theme small 383:387 arg1 set 389:391 a small set 381:391 a small set of most likely glycan structures 381:424 Even though MALDI-TOF profiles give only intact masses without any fragmentation information, expert knowledge of the most common glycans and biosynthetic pathways in the biological system can point to a small set of most likely glycan structures at the "cartoon" level of detail. 26047718 0 56 theme N-glycan 53:60 arg1 profiles 62:69 MALDI-TOF N-glycan profiles 43:69 MALDI-TOF N-glycan profiles 43:69 Benchmark study of automatic annotation of MALDI-TOF N-glycan profiles. 26047718 6 57 theme cases 1064:1068 arg1 % 1055:1055 over 90% 1048:1055 over 90% of all cases 1048:1068 On peaks for which both Cartoonist and the expert give unique cartoons, the two cartoons agree in over 90% of all cases. 26047718 6 57 theme cases 1064:1068 arg1 cases 1064:1068 all cases 1060:1068 all cases 1060:1068 On peaks for which both Cartoonist and the expert give unique cartoons, the two cartoons agree in over 90% of all cases. 26047718 4 58 theme N-glycan 664:671 arg1 data 673:676 human and mouse N-glycan data 648:676 human and mouse N-glycan data from the Consortium for Functional Glycomics 648:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 26047718 4 59 theme automatic 590:598 arg1 annotations 600:610 Cartoonist's automatic annotations 577:610 Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics 577:721 Here we benchmark Cartoonist's automatic annotations against human expert annotations on human and mouse N-glycan data from the Consortium for Functional Glycomics. 25118826 4 0 theme sodium 912:917 arg1 reduction 931:939 sodium borohydride reduction 912:939 sodium borohydride reduction 912:939 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 2 1 theme data 591:594 arg1 analysis 596:603 data analysis 591:603 data analysis 591:603 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 8 2 theme Glycomics 1752:1760 arg1 Quantification 1785:1798 the Glycomics Quintavariate Informed Quantification 1748:1798 the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al 1748:1846 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 2 theme Glycomics 1752:1760 arg1 Kronewitter 1830:1840 Kronewitter 1830:1840 Kronewitter 1830:1840 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 2 theme Glycomics 1752:1760 arg1 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 1 3 gly glycoproteins 202:214 arg1 glycoproteins 202:214 human serum glycoproteins 190:214 human serum glycoproteins 190:214 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 5 4 from desalting 1056:1064 arg1 column 1119:1124 the analytical column 1104:1124 the analytical column 1104:1124 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 12 5 theme initial 2097:2103 arg1 observations 2105:2116 our initial observations 2093:2116 our initial observations 2093:2116 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 7 6 theme gentler 1561:1567 arg1 conditions 1580:1589 gentler ionization conditions 1561:1589 gentler ionization conditions 1561:1589 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 3 7 theme glycan 716:721 arg1 characterization 723:738 ESI-MS glycan characterization 709:738 ESI-MS glycan characterization 709:738 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 1 8 theme serum 196:200 arg1 glycoproteins 202:214 human serum glycoproteins 190:214 human serum glycoproteins 190:214 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 12 9 theme glycome 2070:2076 arg1 context 2043:2049 the context 2039:2049 the context of the human serum glycome 2039:2076 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 2 10 theme software 572:579 arg1 tools 581:585 software tools 572:585 software tools for data analysis 572:603 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 6 11 theme additional 1412:1421 arg1 power 1433:1437 additional resolving power 1412:1437 additional resolving power 1412:1437 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 4 12 theme quantitative 978:989 arg1 yields 991:996 quantitative yields 978:996 quantitative yields 978:996 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 11 13 theme glycan 1919:1924 arg1 sensitivity 1936:1946 glycan detection sensitivity 1919:1946 glycan detection sensitivity 1919:1946 2014, 86, 6268-6276), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. 25118826 1 14 theme human 227:231 arg1 N-glycome 245:253 the human blood serum N-glycome 223:253 the human blood serum N-glycome 223:253 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 12 15 theme human 2058:2062 arg1 glycome 2070:2076 the human serum glycome 2054:2076 the human serum glycome 2054:2076 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 0 16 from developments 68:79 arg1 spectrometry 150:161 electrospray ionization-mass spectrometry 121:161 electrospray ionization-mass spectrometry 121:161 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 0 16 from developments 68:79 arg1 separations 104:114 separations 104:114 separations 104:114 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 0 16 from developments 68:79 arg1 preparation 91:101 sample preparation 84:101 sample preparation 84:101 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 6 17 theme carbon 1310:1315 arg1 columns 1317:1323 extended length graphitized carbon columns 1282:1323 extended length graphitized carbon columns 1282:1323 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 18 theme phase 1386:1390 arg1 gradient 1392:1399 a formic acid/reversed phase gradient 1363:1399 a formic acid/reversed phase gradient 1363:1399 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 19 theme length 1291:1296 arg1 columns 1317:1323 extended length graphitized carbon columns 1282:1323 extended length graphitized carbon columns 1282:1323 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 12 20 theme N-glycans 2178:2186 arg1 class 2150:2154 a new class 2144:2154 a new class of heavily sialylated N-glycans, including polysialylated N-glycans 2144:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 6 21 theme formic 1365:1370 arg1 gradient 1392:1399 a formic acid/reversed phase gradient 1363:1399 a formic acid/reversed phase gradient 1363:1399 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 12 22 gly polysialylated 2199:2212 arg1 N-glycans 2214:2222 polysialylated N-glycans 2199:2222 polysialylated N-glycans 2199:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 3 23 theme several 621:627 arg1 improvements 629:640 several improvements 621:640 several improvements 621:640 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 1 24 dep both 259:262 arg1 diversity 177:185 The N-glycan diversity 164:185 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, 164:254 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 24 dep both 259:262 arg1 complex 264:270 complex 264:270 complex 264:270 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 24 dep both 259:262 arg1 both 259:262 both 259:262 both 259:262 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 25 theme synthesizable 334:346 arg1 structures 311:320 glycan structures 304:320 glycan structures potentially synthesizable 304:346 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 26 theme N-glycan 168:175 arg1 both 259:262 both 259:262 both 259:262 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 26 theme N-glycan 168:175 arg1 complex 264:270 complex 264:270 complex 264:270 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 26 theme N-glycan 168:175 arg1 diversity 177:185 The N-glycan diversity 164:185 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, 164:254 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 7 27 used utilized 1683:1690 arg2 ionization 1639:1648 subambient pressure ionization 1619:1648 subambient pressure ionization with nanoelectrospray (SPIN) 1619:1677 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 12 28 theme class 2150:2154 arg1 detection 2131:2139 the detection 2127:2139 the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans 2127:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 0 29 theme sample 84:89 arg1 preparation 91:101 sample preparation 84:101 sample preparation 84:101 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 1 30 theme human 351:355 arg1 enzymes 371:377 human glycosylation enzymes 351:377 human glycosylation enzymes 351:377 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 6 31 theme acid 1346:1349 arg1 modifier 1351:1358 an acid modifier 1343:1358 an acid modifier to a formic acid/reversed phase gradient 1343:1399 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 31 theme acid 1346:1349 arg1 TFA 1336:1338 TFA 1336:1338 TFA 1336:1338 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 12 32 from application 2006:2016 arg1 included 2118:2125 included 2118:2125 included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans 2118:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 12 32 from application 2006:2016 arg1 context 2043:2049 the context 2039:2049 the context of the human serum glycome 2039:2076 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 2 33 theme ionization-mass 529:543 arg1 ESI-MS 559:564 ESI-MS 559:564 ESI-MS 559:564 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 2 33 theme ionization-mass 529:543 arg1 spectrometry 545:556 electrospray ionization-mass spectrometry 516:556 electrospray ionization-mass spectrometry (ESI-MS) 516:565 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 4 34 theme preparation 788:798 arg1 improvements 800:811 Sample preparation improvements 781:811 Sample preparation improvements 781:811 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 5 35 theme analyte 1161:1167 arg1 division 1176:1183 analyte signal division 1161:1183 analyte signal division between multiple reducing-end-forms or cation adducts 1161:1237 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 2 36 theme sample 447:452 arg1 preparation 454:464 sample preparation 447:464 sample preparation 447:464 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 0 37 theme Polysialylated 0:13 arg1 N-glycans 15:23 Polysialylated N-glycans 0:23 Polysialylated N-glycans 0:23 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 2 38 theme analytical 477:486 arg1 platforms 488:496 available analytical platforms 467:496 available analytical platforms 467:496 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 4 39 theme F 863:863 arg1 acidified 822:830 acidified 822:830 acidified 822:830 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 4 39 theme F 863:863 arg1 release 874:880 PNGase F N-glycan release 856:880 PNGase F N-glycan release 856:880 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 6 40 theme sialylated 1503:1512 arg1 glycans 1514:1520 both large and heavily sialylated glycans 1480:1520 both large and heavily sialylated glycans 1480:1520 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 0 41 theme human 39:43 arg1 serum 45:49 human serum 39:49 human serum 39:49 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 7 42 theme pressure 1630:1637 arg1 ionization 1639:1648 subambient pressure ionization 1619:1648 subambient pressure ionization with nanoelectrospray (SPIN) 1619:1677 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 5 43 theme solid 1074:1078 arg1 extraction 1086:1095 solid phase extraction 1074:1095 solid phase extraction 1074:1095 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 1 44 theme structures 311:320 arg1 range 295:299 the range 291:299 the range of glycan structures potentially synthesizable 291:346 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 8 45 theme Quintavariate 1762:1774 arg1 Quantification 1785:1798 the Glycomics Quintavariate Informed Quantification 1748:1798 the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al 1748:1846 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 45 theme Quintavariate 1762:1774 arg1 Kronewitter 1830:1840 Kronewitter 1830:1840 Kronewitter 1830:1840 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 45 theme Quintavariate 1762:1774 arg1 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 1 46 theme blood 233:237 arg1 N-glycome 245:253 the human blood serum N-glycome 223:253 the human blood serum N-glycome 223:253 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 7 47 theme source-MS 1598:1606 arg1 interface 1608:1616 the source-MS interface 1594:1616 the source-MS interface 1594:1616 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 5 48 theme Two-stage 1046:1054 arg1 desalting 1056:1064 Two-stage desalting 1046:1064 Two-stage desalting (during solid phase extraction and on the analytical column) 1046:1125 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 8 49 theme improved 1704:1711 arg1 methods 1713:1719 these improved methods 1698:1719 these improved methods 1698:1719 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 3 50 theme sample 667:672 arg1 preparation 674:684 sample preparation 667:684 sample preparation 667:684 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 2 51 theme known 384:388 arg1 glycome 390:396 The known glycome 380:396 The known glycome 380:396 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 5 52 theme cation 1224:1229 arg1 adducts 1231:1237 cation adducts 1224:1237 cation adducts 1224:1237 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 3 53 theme ESI-MS 709:714 arg1 characterization 723:738 ESI-MS glycan characterization 709:738 ESI-MS glycan characterization 709:738 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 11 54 theme glycan 1969:1974 arg1 coverage 1976:1983 expanded glycan coverage 1960:1983 expanded glycan coverage 1960:1983 2014, 86, 6268-6276), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. 25118826 7 55 theme ionization 1569:1578 arg1 conditions 1580:1589 gentler ionization conditions 1561:1589 gentler ionization conditions 1561:1589 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 4 56 theme borohydride 919:929 arg1 reduction 931:939 sodium borohydride reduction 912:939 sodium borohydride reduction 912:939 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 7 57 theme MS 1534:1535 arg1 sensitivity 1537:1547 MS sensitivity 1534:1547 MS sensitivity 1534:1547 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 1 58 theme human 190:194 arg1 glycoproteins 202:214 human serum glycoproteins 190:214 human serum glycoproteins 190:214 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 6 59 theme Online 1240:1245 arg1 separations 1247:1257 Online separations 1240:1257 Online separations 1240:1257 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 12 60 theme serum 2064:2068 arg1 glycome 2070:2076 the human serum glycome 2054:2076 the human serum glycome 2054:2076 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 1 61 theme glycoproteins 202:214 arg1 both 259:262 both 259:262 both 259:262 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 61 theme glycoproteins 202:214 arg1 complex 264:270 complex 264:270 complex 264:270 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 1 61 theme glycoproteins 202:214 arg1 diversity 177:185 The N-glycan diversity 164:185 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, 164:254 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 5 62 theme multiple 1193:1200 arg1 reducing-end-forms 1202:1219 multiple reducing-end-forms 1193:1219 multiple reducing-end-forms 1193:1219 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 6 63 theme resolving 1423:1431 arg1 power 1433:1437 additional resolving power 1412:1437 additional resolving power 1412:1437 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 7 64 with ionization 1639:1648 arg1 SPIN 1673:1676 nanoelectrospray (SPIN) 1655:1677 nanoelectrospray (SPIN) 1655:1677 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 1 65 dep N-glycome 245:253 arg1 i.e. 217:220 i.e. 217:220 i.e. 217:220 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 3 66 theme polysialylated 755:768 arg1 N-glycans 770:778 polysialylated N-glycans 755:778 polysialylated N-glycans 755:778 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 8 67 dep described 1819:1827 arg1 Quantification 1785:1798 the Glycomics Quintavariate Informed Quantification 1748:1798 the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al 1748:1846 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 67 dep described 1819:1827 arg1 Kronewitter 1830:1840 Kronewitter 1830:1840 Kronewitter 1830:1840 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 12 68 theme polysialylated 2199:2212 arg1 N-glycans 2214:2222 polysialylated N-glycans 2199:2222 polysialylated N-glycans 2199:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 11 69 theme detection 1926:1934 arg1 sensitivity 1936:1946 glycan detection sensitivity 1919:1946 glycan detection sensitivity 1919:1946 2014, 86, 6268-6276), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. 25118826 0 70 theme ionization-mass 134:148 arg1 spectrometry 150:161 electrospray ionization-mass spectrometry 121:161 electrospray ionization-mass spectrometry 121:161 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 6 71 theme graphitized 1298:1308 arg1 columns 1317:1323 extended length graphitized carbon columns 1282:1323 extended length graphitized carbon columns 1282:1323 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 72 theme acid/reversed 1372:1384 arg1 gradient 1392:1399 a formic acid/reversed phase gradient 1363:1399 a formic acid/reversed phase gradient 1363:1399 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 73 theme extended 1282:1289 arg1 columns 1317:1323 extended length graphitized carbon columns 1282:1323 extended length graphitized carbon columns 1282:1323 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 12 74 theme sialylated 2167:2176 arg1 N-glycans 2214:2222 polysialylated N-glycans 2199:2222 polysialylated N-glycans 2199:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 12 74 theme sialylated 2167:2176 arg1 N-glycans 2178:2186 heavily sialylated N-glycans 2159:2186 heavily sialylated N-glycans 2159:2186 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 6 75 theme large 1485:1489 arg1 glycans 1514:1520 both large and heavily sialylated glycans 1480:1520 both large and heavily sialylated glycans 1480:1520 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 5 76 theme analytical 1108:1117 arg1 column 1119:1124 the analytical column 1104:1124 the analytical column 1104:1124 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 11 77 dep able 1890:1893 arg1 6268-6276 1871:1879 6268-6276 1871:1879 6268-6276 1871:1879 2014, 86, 6268-6276), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. 25118826 12 78 theme advances 2027:2034 arg1 application 2006:2016 the application 2002:2016 the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans 2002:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 4 79 theme glycoforms 1025:1034 arg1 number 1015:1020 the number 1011:1020 the number of glycoforms detected 1011:1043 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 12 80 theme new 2146:2148 arg1 class 2150:2154 a new class 2144:2154 a new class of heavily sialylated N-glycans, including polysialylated N-glycans 2144:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 6 81 theme improved 1457:1464 arg1 desorption 1466:1475 significantly improved desorption 1443:1475 significantly improved desorption of both large and heavily sialylated glycans 1443:1520 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 1 82 theme glycosylation 357:369 arg1 enzymes 371:377 human glycosylation enzymes 351:377 human glycosylation enzymes 351:377 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 6 83 gly sialylated 1503:1512 arg1 glycans 1514:1520 both large and heavily sialylated glycans 1480:1520 both large and heavily sialylated glycans 1480:1520 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 2 84 theme electrospray 516:527 arg1 ESI-MS 559:564 ESI-MS 559:564 ESI-MS 559:564 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 2 84 theme electrospray 516:527 arg1 spectrometry 545:556 electrospray ionization-mass spectrometry 516:556 electrospray ionization-mass spectrometry (ESI-MS) 516:565 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 4 85 theme Sample 781:786 arg1 improvements 800:811 Sample preparation improvements 781:811 Sample preparation improvements 781:811 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 12 86 gly sialylated 2167:2176 arg1 N-glycans 2214:2222 polysialylated N-glycans 2199:2222 polysialylated N-glycans 2199:2222 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 12 86 gly sialylated 2167:2176 arg1 N-glycans 2178:2186 heavily sialylated N-glycans 2159:2186 heavily sialylated N-glycans 2159:2186 We demonstrated the application of these advances in the context of the human serum glycome, and for which our initial observations included the detection of a new class of heavily sialylated N-glycans, including polysialylated N-glycans. 25118826 3 87 gly polysialylated 755:768 arg1 N-glycans 770:778 polysialylated N-glycans 755:778 polysialylated N-glycans 755:778 In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans. 25118826 5 88 theme signal 1169:1174 arg1 division 1176:1183 analyte signal division 1161:1183 analyte signal division between multiple reducing-end-forms or cation adducts 1161:1237 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 8 89 dep Kronewitter 1830:1840 arg1 al 1845:1846 Kronewitter et al 1830:1846 Kronewitter et al 1830:1846 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 6 90 theme glycans 1514:1520 arg1 power 1433:1437 additional resolving power 1412:1437 additional resolving power 1412:1437 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 6 90 theme glycans 1514:1520 arg1 desorption 1466:1475 significantly improved desorption 1443:1475 significantly improved desorption of both large and heavily sialylated glycans 1443:1520 Online separations were improved by using extended length graphitized carbon columns and adding TFA as an acid modifier to a formic acid/reversed phase gradient, providing additional resolving power and significantly improved desorption of both large and heavily sialylated glycans. 25118826 7 91 theme nanoelectrospray 1655:1670 arg1 SPIN 1673:1676 nanoelectrospray (SPIN) 1655:1677 nanoelectrospray (SPIN) 1655:1677 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 0 92 gly Polysialylated 0:13 arg1 N-glycans 15:23 Polysialylated N-glycans 0:23 Polysialylated N-glycans 0:23 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 2 93 theme available 467:475 arg1 platforms 488:496 available analytical platforms 467:496 available analytical platforms 467:496 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 4 94 theme PNGase 856:861 arg1 F 863:863 PNGase F 856:863 PNGase F N-glycan release 856:880 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 0 95 theme combined 59:66 arg1 developments 68:79 combined developments 59:79 combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry 59:161 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 25118826 1 96 theme glycan 304:309 arg1 structures 311:320 glycan structures 304:320 glycan structures potentially synthesizable 304:346 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 5 97 theme phase 1080:1084 arg1 extraction 1086:1095 solid phase extraction 1074:1095 solid phase extraction 1074:1095 Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division between multiple reducing-end-forms or cation adducts. 25118826 2 98 theme preparation 454:464 arg1 platforms 488:496 available analytical platforms 467:496 available analytical platforms 467:496 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 2 98 theme preparation 454:464 arg1 methods 436:442 methods 436:442 methods of sample preparation 436:464 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 2 98 theme preparation 454:464 arg1 tools 581:585 software tools 572:585 software tools for data analysis 572:603 The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, e.g., based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis. 25118826 4 99 theme N-glycan 865:872 arg1 acidified 822:830 acidified 822:830 acidified 822:830 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 4 99 theme N-glycan 865:872 arg1 release 874:880 PNGase F N-glycan release 856:880 PNGase F N-glycan release 856:880 Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected. 25118826 11 100 theme expanded 1960:1967 arg1 coverage 1976:1983 expanded glycan coverage 1960:1983 expanded glycan coverage 1960:1983 2014, 86, 6268-6276), we are able to significantly extend glycan detection sensitivity and provide expanded glycan coverage. 25118826 8 101 theme Informed 1776:1783 arg1 Quantification 1785:1798 the Glycomics Quintavariate Informed Quantification 1748:1798 the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al 1748:1846 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 101 theme Informed 1776:1783 arg1 Kronewitter 1830:1840 Kronewitter 1830:1840 Kronewitter 1830:1840 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 8 101 theme Informed 1776:1783 arg1 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 GlyQ-IQ 1801:1807 When these improved methods are combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described (Kronewitter et al. 25118826 7 102 theme subambient 1619:1628 arg1 ionization 1639:1648 subambient pressure ionization 1619:1648 subambient pressure ionization with nanoelectrospray (SPIN) 1619:1677 To improve MS sensitivity and provide gentler ionization conditions at the source-MS interface, subambient pressure ionization with nanoelectrospray (SPIN) was utilized. 25118826 1 103 theme serum 239:243 arg1 N-glycome 245:253 the human blood serum N-glycome 223:253 the human blood serum N-glycome 223:253 The N-glycan diversity of human serum glycoproteins, i.e., the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes. 25118826 0 104 theme electrospray 121:132 arg1 spectrometry 150:161 electrospray ionization-mass spectrometry 121:161 electrospray ionization-mass spectrometry 121:161 Polysialylated N-glycans identified in human serum through combined developments in sample preparation, separations, and electrospray ionization-mass spectrometry. 26587982 4 0 from patients 655:662 arg1 sera 583:586 sera 583:586 sera from experimentally infected mice and from LASV-convalescent human patients 583:662 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 7 1 with studies 998:1004 arg1 antibodies 1022:1031 monoclonal antibodies 1011:1031 monoclonal antibodies 1011:1031 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 1 2 theme hemorrhagic 147:157 arg1 fever 159:163 severe hemorrhagic fever 140:163 severe hemorrhagic fever in humans 140:173 Arenaviruses such as Lassa virus (LASV) can cause severe hemorrhagic fever in humans. 26587982 10 3 theme convalescent 1550:1561 arg1 therapy 1569:1575 convalescent serum therapy 1550:1575 convalescent serum therapy 1550:1575 This immune evasion mechanism imposes limitations on antibody-based vaccination and convalescent serum therapy. 26587982 8 4 theme ensuing 1268:1274 arg1 response 1285:1292 the ensuing antibody response 1264:1292 the ensuing antibody response 1264:1292 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 3 5 dep engineered 449:458 arg1 investigate 385:395 investigate 385:395 To investigate the mechanisms underlying arenavirus nAb evasion 382:444 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 8 6 theme infected 1146:1153 arg1 mice 1155:1158 infected mice 1146:1158 infected mice 1146:1158 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 1 7 from fever 159:163 arg1 humans 168:173 humans 168:173 humans 168:173 Arenaviruses such as Lassa virus (LASV) can cause severe hemorrhagic fever in humans. 26587982 4 8 theme LASV-convalescent 631:647 arg1 patients 655:662 LASV-convalescent human patients 631:662 LASV-convalescent human patients 631:662 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 5 9 theme globular 786:793 arg1 head 795:798 the arenavirus envelope protein's globular head 752:798 the arenavirus envelope protein's globular head 752:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 9 10 theme efficient 1423:1431 arg1 control 1457:1463 efficient antibody-mediated virus control 1423:1463 efficient antibody-mediated virus control 1423:1463 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 10 11 theme immune 1471:1476 arg1 evasion 1478:1484 immune evasion 1471:1484 This immune evasion mechanism 1466:1494 This immune evasion mechanism imposes limitations on antibody-based vaccination and convalescent serum therapy. 26587982 9 12 theme antibody-mediated 1433:1449 arg1 control 1457:1463 efficient antibody-mediated virus control 1423:1463 efficient antibody-mediated virus control 1423:1463 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 5 13 theme response 669:676 arg1 kinetics 678:685 NAb response kinetics 665:685 NAb response kinetics in mice 665:693 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 5 14 theme NAb 665:667 arg1 kinetics 678:685 NAb response kinetics 665:685 NAb response kinetics in mice 665:693 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 3 15 theme nAb 434:436 arg1 evasion 438:444 arenavirus nAb evasion 423:444 arenavirus nAb evasion 423:444 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 4 16 from mice 617:620 arg1 sera 583:586 sera 583:586 sera from experimentally infected mice and from LASV-convalescent human patients 583:662 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 9 17 theme virus 1451:1455 arg1 control 1457:1463 efficient antibody-mediated virus control 1423:1463 efficient antibody-mediated virus control 1423:1463 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 6 18 with infection 837:845 arg1 viruses 871:877 fully glycosylated viruses 852:877 fully glycosylated viruses 852:877 Additionally and most intriguingly, infection with fully glycosylated viruses elicited antibodies, which neutralized predominantly their glycan-deficient variants, both in mice and humans. 26587982 9 19 theme envelope 1312:1319 arg1 glycosylation 1321:1333 arenavirus envelope glycosylation 1301:1333 arenavirus envelope glycosylation 1301:1333 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 5 20 from kinetics 678:685 arg1 mice 690:693 mice 690:693 mice 690:693 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 8 21 theme envelope 1165:1172 arg1 glycan 1174:1179 the envelope glycan 1161:1179 the envelope glycan shield 1161:1186 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 8 22 theme viral 1208:1212 arg1 infection 1214:1222 protracted viral infection 1197:1222 protracted viral infection 1197:1222 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 0 23 theme Arenavirus 0:9 arg1 Glycan 11:16 Arenavirus Glycan Shield 0:23 Arenavirus Glycan Shield 0:23 Arenavirus Glycan Shield Promotes Neutralizing Antibody Evasion and Protracted Infection. 26587982 10 24 theme serum 1563:1567 arg1 therapy 1569:1575 convalescent serum therapy 1550:1575 convalescent serum therapy 1550:1575 This immune evasion mechanism imposes limitations on antibody-based vaccination and convalescent serum therapy. 26587982 2 25 theme unifying 289:296 arg1 impediment 187:196 a major impediment 179:196 a major impediment to vaccine development 179:219 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 25 theme unifying 289:296 arg1 characteristic 298:311 a unifying characteristic 287:311 a unifying characteristic of both natural infection and all vaccine candidates tested to date 287:379 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 3 26 theme arenavirus 423:432 arg1 evasion 438:444 arenavirus nAb evasion 423:444 arenavirus nAb evasion 423:444 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 6 27 theme glycosylated 858:869 arg1 viruses 871:877 fully glycosylated viruses 852:877 fully glycosylated viruses 852:877 Additionally and most intriguingly, infection with fully glycosylated viruses elicited antibodies, which neutralized predominantly their glycan-deficient variants, both in mice and humans. 26587982 1 28 theme Lassa 111:115 arg1 LASV 124:127 LASV 124:127 LASV 124:127 Arenaviruses such as Lassa virus (LASV) can cause severe hemorrhagic fever in humans. 26587982 1 28 theme Lassa 111:115 arg1 virus 117:121 Lassa virus 111:121 Lassa virus (LASV) 111:128 Arenaviruses such as Lassa virus (LASV) can cause severe hemorrhagic fever in humans. 26587982 5 29 link N-linked 725:732 arg1 density 741:747 the N-linked glycan density 721:747 the N-linked glycan density in the arenavirus envelope protein's globular head 721:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 3 30 theme several 460:466 arg1 viruses 497:503 several arenavirus envelope-chimeric viruses 460:503 several arenavirus envelope-chimeric viruses 460:503 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 0 31 theme Shield 18:23 arg1 Glycan 11:16 Arenavirus Glycan Shield 0:23 Arenavirus Glycan Shield 0:23 Arenavirus Glycan Shield Promotes Neutralizing Antibody Evasion and Protracted Infection. 26587982 4 32 theme infected 608:615 arg1 mice 617:620 experimentally infected mice 593:620 experimentally infected mice 593:620 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 5 33 theme arenavirus 756:765 arg1 protein 776:782 the arenavirus envelope protein's 752:784 the arenavirus envelope protein's globular head 752:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 9 34 theme protective 1347:1356 arg1 efficacy 1358:1365 the protective efficacy 1343:1365 the protective efficacy rather than the induction of nAbs 1343:1399 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 7 35 theme envelope 1048:1055 arg1 glycans 1057:1063 envelope glycans 1048:1063 envelope glycans 1048:1063 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 3 36 theme arenavirus 468:477 arg1 viruses 497:503 several arenavirus envelope-chimeric viruses 460:503 several arenavirus envelope-chimeric viruses 460:503 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 0 37 theme Antibody 47:54 arg1 Evasion 56:62 Neutralizing Antibody Evasion 34:62 Neutralizing Antibody Evasion 34:62 Arenavirus Glycan Shield Promotes Neutralizing Antibody Evasion and Protracted Infection. 26587982 3 38 theme glycan-deficient 509:524 arg1 variants 526:533 glycan-deficient variants 509:533 glycan-deficient variants thereof 509:541 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 5 39 theme envelope 767:774 arg1 protein 776:782 the arenavirus envelope protein's 752:784 the arenavirus envelope protein's globular head 752:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 2 40 theme delayed 222:228 arg1 nAb 262:264 nAb 262:264 nAb 262:264 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 40 theme delayed 222:228 arg1 antibody 252:259 delayed and weak neutralizing antibody 222:259 delayed and weak neutralizing antibody (nAb) responses 222:275 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 0 41 theme Neutralizing 34:45 arg1 Evasion 56:62 Neutralizing Antibody Evasion 34:62 Neutralizing Antibody Evasion 34:62 Arenavirus Glycan Shield Promotes Neutralizing Antibody Evasion and Protracted Infection. 26587982 4 42 theme neutralization 557:570 arg1 tests 572:576 neutralization tests 557:576 neutralization tests 557:576 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 6 43 theme glycan-deficient 938:953 arg1 both 965:968 both 965:968 both 965:968 Additionally and most intriguingly, infection with fully glycosylated viruses elicited antibodies, which neutralized predominantly their glycan-deficient variants, both in mice and humans. 26587982 6 43 theme glycan-deficient 938:953 arg1 variants 955:962 their glycan-deficient variants 932:962 their glycan-deficient variants 932:962 Additionally and most intriguingly, infection with fully glycosylated viruses elicited antibodies, which neutralized predominantly their glycan-deficient variants, both in mice and humans. 26587982 10 44 theme evasion 1478:1484 arg1 mechanism 1486:1494 This immune evasion mechanism 1466:1494 This immune evasion mechanism 1466:1494 This immune evasion mechanism imposes limitations on antibody-based vaccination and convalescent serum therapy. 26587982 8 45 theme antibody 1276:1283 arg1 response 1285:1292 the ensuing antibody response 1264:1292 the ensuing antibody response 1264:1292 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 5 46 theme N-linked 725:732 arg1 density 741:747 the N-linked glycan density 721:747 the N-linked glycan density in the arenavirus envelope protein's globular head 721:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 7 47 theme virus 1121:1125 arg1 neutralization 1127:1140 virus neutralization 1121:1140 virus neutralization 1121:1140 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 2 48 theme infection 329:337 arg1 impediment 187:196 a major impediment 179:196 a major impediment to vaccine development 179:219 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 48 theme infection 329:337 arg1 characteristic 298:311 a unifying characteristic 287:311 a unifying characteristic of both natural infection and all vaccine candidates tested to date 287:379 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 8 49 dep glycan 1174:1179 arg1 shield 1181:1186 shield 1181:1186 shield 1181:1186 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 6 50 gly glycosylated 858:869 arg1 viruses 871:877 fully glycosylated viruses 852:877 fully glycosylated viruses 852:877 Additionally and most intriguingly, infection with fully glycosylated viruses elicited antibodies, which neutralized predominantly their glycan-deficient variants, both in mice and humans. 26587982 5 51 theme glycan 734:739 arg1 density 741:747 the N-linked glycan density 721:747 the N-linked glycan density in the arenavirus envelope protein's globular head 721:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 2 52 theme vaccine 201:207 arg1 development 209:219 vaccine development 201:219 vaccine development 201:219 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 53 theme natural 321:327 arg1 infection 329:337 natural infection 321:337 natural infection 321:337 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 7 54 theme Binding 990:996 arg1 studies 998:1004 Binding studies 990:1004 Binding studies with monoclonal antibodies 990:1031 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 7 55 theme monoclonal 1011:1020 arg1 antibodies 1022:1031 monoclonal antibodies 1011:1031 monoclonal antibodies 1011:1031 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 8 56 theme protracted 1197:1206 arg1 infection 1214:1222 protracted viral infection 1197:1222 protracted viral infection 1197:1222 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 2 57 theme antibody 252:259 arg1 responses 267:275 delayed and weak neutralizing antibody (nAb) responses 222:275 delayed and weak neutralizing antibody (nAb) responses 222:275 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 3 58 theme envelope-chimeric 479:495 arg1 viruses 497:503 several arenavirus envelope-chimeric viruses 460:503 several arenavirus envelope-chimeric viruses 460:503 To investigate the mechanisms underlying arenavirus nAb evasion we engineered several arenavirus envelope-chimeric viruses and glycan-deficient variants thereof. 26587982 5 59 from density 741:747 arg1 head 795:798 the arenavirus envelope protein's globular head 752:798 the arenavirus envelope protein's globular head 752:798 NAb response kinetics in mice correlated inversely with the N-linked glycan density in the arenavirus envelope protein's globular head. 26587982 2 60 theme major 181:185 arg1 impediment 187:196 a major impediment 179:196 a major impediment to vaccine development 179:219 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 60 theme major 181:185 arg1 characteristic 298:311 a unifying characteristic 287:311 a unifying characteristic of both natural infection and all vaccine candidates tested to date 287:379 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 61 theme neutralizing 239:250 arg1 nAb 262:264 nAb 262:264 nAb 262:264 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 61 theme neutralizing 239:250 arg1 antibody 252:259 delayed and weak neutralizing antibody 222:259 delayed and weak neutralizing antibody (nAb) responses 222:275 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 1 62 theme severe 140:145 arg1 fever 159:163 severe hemorrhagic fever 140:163 severe hemorrhagic fever in humans 140:173 Arenaviruses such as Lassa virus (LASV) can cause severe hemorrhagic fever in humans. 26587982 9 63 theme arenavirus 1301:1310 arg1 glycosylation 1321:1333 arenavirus envelope glycosylation 1301:1333 arenavirus envelope glycosylation 1301:1333 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 4 64 theme human 649:653 arg1 patients 655:662 LASV-convalescent human patients 631:662 LASV-convalescent human patients 631:662 We performed neutralization tests with sera from experimentally infected mice and from LASV-convalescent human patients. 26587982 8 65 theme timely 1242:1247 arg1 elimination 1249:1259 its timely elimination 1238:1259 its timely elimination by the ensuing antibody response 1238:1292 In infected mice, the envelope glycan shield promoted protracted viral infection by preventing its timely elimination by the ensuing antibody response. 26587982 9 66 theme nAbs 1396:1399 arg1 induction 1383:1391 the induction 1379:1391 the protective efficacy rather than the induction of nAbs 1343:1399 Thus, arenavirus envelope glycosylation impairs the protective efficacy rather than the induction of nAbs, and thereby prevents efficient antibody-mediated virus control. 26587982 2 67 theme weak 234:237 arg1 nAb 262:264 nAb 262:264 nAb 262:264 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 67 theme weak 234:237 arg1 antibody 252:259 delayed and weak neutralizing antibody 222:259 delayed and weak neutralizing antibody (nAb) responses 222:275 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 68 theme candidates 355:364 arg1 impediment 187:196 a major impediment 179:196 a major impediment to vaccine development 179:219 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 2 68 theme candidates 355:364 arg1 characteristic 298:311 a unifying characteristic 287:311 a unifying characteristic of both natural infection and all vaccine candidates tested to date 287:379 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 26587982 10 69 theme antibody-based 1519:1532 arg1 vaccination 1534:1544 antibody-based vaccination 1519:1544 antibody-based vaccination 1519:1544 This immune evasion mechanism imposes limitations on antibody-based vaccination and convalescent serum therapy. 26587982 0 70 theme Protracted 68:77 arg1 Infection 79:87 Protracted Infection 68:87 Protracted Infection 68:87 Arenavirus Glycan Shield Promotes Neutralizing Antibody Evasion and Protracted Infection. 26587982 7 71 theme nAb 1073:1075 arg1 occupancy 1086:1094 occupancy 1086:1094 occupancy 1086:1094 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 7 71 theme nAb 1073:1075 arg1 on-rate 1077:1083 nAb on-rate 1073:1083 nAb on-rate 1073:1083 Binding studies with monoclonal antibodies indicated that envelope glycans reduced nAb on-rate, occupancy and thereby counteracted virus neutralization. 26587982 2 72 theme vaccine 347:353 arg1 candidates 355:364 all vaccine candidates 343:364 all vaccine candidates tested to date 343:379 As a major impediment to vaccine development, delayed and weak neutralizing antibody (nAb) responses represent a unifying characteristic of both natural infection and all vaccine candidates tested to date. 25733616 7 0 theme 2-sugar 1328:1334 arg1 glycan 1336:1341 a 2-sugar glycan 1326:1341 a 2-sugar glycan 1326:1341 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 14 1 theme bacterial 2627:2635 arg1 gene 2637:2640 a bacterial gene 2625:2640 a bacterial gene in a complex sugar biosynthesis pathway 2625:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 12 2 mod modified 2267:2274 arg1 Pilins 2256:2261 Pilins 2256:2261 Pilins 2256:2261 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 12 2 mod modified 2267:2274 arg3 N-glycan 2293:2300 a different N-glycan 2281:2300 a different N-glycan consisting of the archaellin tetrasaccharide 2281:2345 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 12 2 mod modified 2267:2274 arg3 hexose 2370:2375 an additional hexose 2356:2375 an additional hexose attached to the linking sugar 2356:2405 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 2 3 mod modified 379:386 arg3 tetrasaccharide 405:419 an N-linked tetrasaccharide 393:419 an N-linked tetrasaccharide 393:419 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 2 3 mod modified 379:386 arg1 Archaellins 363:373 Archaellins 363:373 Archaellins 363:373 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 2 4 with tetrasaccharide 405:419 arg1 Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 440:495 the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 426:495 the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 426:495 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 9 5 theme step 1800:1803 arg1 complementation 1771:1785 Cross-domain complementation 1758:1785 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 1758:1845 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 10 6 theme third 2046:2050 arg1 sugars 2052:2057 the second and third sugars 2031:2057 the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2031:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 1 7 gly glycoprotein 340:351 arg1 glycoprotein 340:351 glycoprotein subunits 340:360 glycoprotein subunits 340:360 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 14 8 theme gene 2597:2600 arg1 example 2574:2580 a rare example 2567:2580 a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway 2567:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 12 9 theme different 2283:2291 arg1 N-glycan 2293:2300 a different N-glycan 2281:2300 a different N-glycan consisting of the archaellin tetrasaccharide 2281:2345 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 0 10 theme di-N-acetylated 202:216 arg1 sugar 218:222 a di-N-acetylated sugar 200:222 a di-N-acetylated sugar 200:222 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 6 11 theme sugar 1243:1247 arg1 biosynthesis 1216:1227 biosynthesis 1216:1227 biosynthesis of the second sugar (GlcNAc3NAcA) 1216:1261 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 10 12 theme N-linked 2066:2073 arg1 tetrasaccharide 2075:2089 the N-linked tetrasaccharide 2062:2089 the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2062:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 4 13 from operons 721:727 arg1 located 672:678 located 672:678 located 672:678 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 13 14 theme mutant 2454:2459 arg1 strain 2461:2466 one mutant strain 2450:2466 one mutant strain 2450:2466 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 6 15 theme Mass 1059:1062 arg1 analysis 1077:1084 Mass spectroscopy analysis 1059:1084 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain 1059:1126 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 7 16 theme sugar 1460:1464 arg1 biosynthesis 1434:1445 biosynthesis 1434:1445 biosynthesis of the third sugar (ManNAc3NAmA6Thr) 1434:1482 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 4 17 from located 672:678 arg1 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 17 from located 672:678 arg1 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 17 from located 672:678 arg1 operons 721:727 two adjacent, divergently transcribed operons 683:727 two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) 683:765 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 0 18 from sugars 51:56 arg1 archaeon 99:106 the archaeon Methanococcus maripaludis 95:132 the archaeon Methanococcus maripaludis 95:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 4 19 dep adjacent 687:694 arg1 transcribed 709:719 transcribed 709:719 transcribed 709:719 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 5 20 contain carrying 885:892 arg2 deletions 894:902 deletions 894:902 deletions in mmp0350, mmp0351, mmp0352, or mmp0353 894:943 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 5 20 contain carrying 885:892 arg1 Mutants 877:883 Mutants 877:883 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 877:943 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 0 21 theme sugars 51:56 arg1 biosynthesis 14:25 biosynthesis 14:25 biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 14:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 1 22 contain has 262:264 arg2 appendages 278:287 two surface appendages 266:287 two surface appendages 266:287 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 22 contain has 262:264 arg2 pili 312:315 type IV pili 304:315 type IV pili 304:315 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 22 contain has 262:264 arg1 maripaludis 250:260 UNLABELLED Methanococcus maripaludis 225:260 UNLABELLED Methanococcus maripaludis 225:260 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 22 contain has 262:264 arg2 archaella 290:298 archaella 290:298 archaella 290:298 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 13 23 theme Mass 2408:2411 arg1 analysis 2426:2433 Mass spectrometry analysis 2408:2433 Mass spectrometry analysis of the pili of one mutant strain 2408:2466 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 14 24 from gene 2637:2640 arg1 pathway 2674:2680 a complex sugar biosynthesis pathway 2645:2680 a complex sugar biosynthesis pathway 2645:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 5 25 theme 1-sugar 1010:1016 arg1 glycan 1018:1023 a 1-sugar glycan 1008:1023 a 1-sugar glycan 1008:1023 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 4 26 theme sugars 869:874 arg1 biosynthesis 844:855 biosynthesis 844:855 biosynthesis of N-glycan sugars 844:874 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 0 27 theme Tetrasaccharide 76:90 arg1 sugars 51:56 the second and third sugars 30:56 the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 30:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 11 28 theme posttranslational 2152:2168 arg1 modification 2170:2181 This posttranslational modification 2147:2181 This posttranslational modification of archaellins 2147:2196 This posttranslational modification of archaellins is important, as it is necessary for archaellum assembly. 25733616 6 29 theme pili 1089:1092 arg1 analysis 1077:1084 Mass spectroscopy analysis 1059:1084 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain 1059:1126 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 2 30 theme N-linked 396:403 arg1 tetrasaccharide 405:419 an N-linked tetrasaccharide 393:419 an N-linked tetrasaccharide 393:419 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 8 31 theme Pseudomonas 1594:1604 arg1 wbpABEDI 1617:1624 Pseudomonas aeruginosa wbpABEDI 1594:1624 Pseudomonas aeruginosa wbpABEDI 1594:1624 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 12 32 theme additional 2359:2368 arg1 hexose 2370:2375 an additional hexose 2356:2375 an additional hexose attached to the linking sugar 2356:2405 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 7 33 theme Δmmp0357 1268:1275 arg1 mutant 1277:1282 The Δmmp0357 mutant 1264:1282 The Δmmp0357 mutant 1264:1282 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 0 34 from Tetrasaccharide 76:90 arg1 archaeon 99:106 the archaeon Methanococcus maripaludis 95:132 the archaeon Methanococcus maripaludis 95:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 5 35 theme Western 1044:1050 arg1 blots 1052:1056 Western blots 1044:1056 Western blots 1044:1056 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 2 36 theme structure 430:438 arg1 Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 440:495 the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 426:495 the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc 426:495 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 3 37 attach attached 631:638 arg2 hexose 624:629 an additional hexose 610:629 an additional hexose attached to GalNAc 610:648 The pilin glycan has an additional hexose attached to GalNAc. 25733616 3 37 attach attached 631:638 arg1 GalNAc 643:648 GalNAc 643:648 GalNAc 643:648 The pilin glycan has an additional hexose attached to GalNAc. 25733616 6 38 theme only 1152:1155 arg1 GalNAc 1157:1162 only GalNAc 1152:1162 only GalNAc 1152:1162 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 9 39 dep P. 1812:1813 arg1 aeruginosa 1815:1824 aeruginosa 1815:1824 aeruginosa 1815:1824 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 1 40 theme IV 309:310 arg1 pili 312:315 type IV pili 304:315 type IV pili 304:315 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 40 theme IV 309:310 arg1 appendages 278:287 two surface appendages 266:287 two surface appendages 266:287 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 7 41 theme purified 1381:1388 arg1 archaella 1390:1398 purified archaella 1381:1398 purified archaella 1381:1398 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 8 42 dep mmp0353 1527:1533 arg1 maripaludis 1488:1498 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 1485:1546 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 1485:1546 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 12 43 theme archaellin 2320:2329 arg1 tetrasaccharide 2331:2345 the archaellin tetrasaccharide 2316:2345 the archaellin tetrasaccharide 2316:2345 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 14 44 theme sugar 2655:2659 arg1 pathway 2674:2680 a complex sugar biosynthesis pathway 2645:2680 a complex sugar biosynthesis pathway 2645:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 9 45 theme Cross-domain 1758:1769 arg1 complementation 1771:1785 Cross-domain complementation 1758:1785 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 1758:1845 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 7 46 from role 1414:1417 arg1 biosynthesis 1434:1445 biosynthesis 1434:1445 biosynthesis of the third sugar (ManNAc3NAmA6Thr) 1434:1482 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 9 47 with complementation 1771:1785 arg1 mmp0357 1839:1845 mmp0357 1839:1845 mmp0357 1839:1845 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 14 48 theme biosynthesis 2661:2672 arg1 pathway 2674:2680 a complex sugar biosynthesis pathway 2645:2680 a complex sugar biosynthesis pathway 2645:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 8 49 theme antigen 1743:1749 arg1 sugar 1751:1755 an O5-specific antigen sugar 1728:1755 an O5-specific antigen sugar 1728:1755 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 8 49 theme antigen 1743:1749 arg1 acid 1722:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 10 50 link N-linked 2066:2073 arg1 tetrasaccharide 2075:2089 the N-linked tetrasaccharide 2062:2089 the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2062:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 1 51 theme surface 270:276 arg1 pili 312:315 type IV pili 304:315 type IV pili 304:315 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 51 theme surface 270:276 arg1 archaella 290:298 archaella 290:298 archaella 290:298 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 51 theme surface 270:276 arg1 appendages 278:287 two surface appendages 266:287 two surface appendages 266:287 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 11 52 theme archaellins 2186:2196 arg1 modification 2170:2181 This posttranslational modification 2147:2181 This posttranslational modification of archaellins 2147:2196 This posttranslational modification of archaellins is important, as it is necessary for archaellum assembly. 25733616 9 53 theme final 1794:1798 arg1 step 1800:1803 the final step 1790:1803 the final step of the P. aeruginosa pathway 1790:1832 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 4 54 theme located 672:678 arg1 genes 666:670 genes 666:670 genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) 666:765 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 10 55 theme sugars 2052:2057 arg1 generation 2017:2026 generation 2017:2026 generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2017:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 10 56 theme genes 1917:1921 arg1 series 1907:1912 a series 1905:1912 a series of genes 1905:1921 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 9 57 theme P. 1812:1813 arg1 pathway 1826:1832 the P. aeruginosa pathway 1808:1832 the P. aeruginosa pathway 1808:1832 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 10 58 theme tetrasaccharide 2075:2089 arg1 sugars 2052:2057 the second and third sugars 2031:2057 the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2031:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 6 59 theme spectroscopy 1064:1075 arg1 analysis 1077:1084 Mass spectroscopy analysis 1059:1084 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain 1059:1126 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 9 60 theme pathway 1826:1832 arg1 step 1800:1803 the final step 1790:1803 the final step of the P. aeruginosa pathway 1790:1832 Cross-domain complementation of the final step of the P. aeruginosa pathway with mmp0357 supports this hypothesis. 25733616 6 61 theme second 1236:1241 arg1 GlcNAc3NAcA 1250:1260 GlcNAc3NAcA 1250:1260 GlcNAc3NAcA 1250:1260 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 6 61 theme second 1236:1241 arg1 sugar 1243:1247 the second sugar 1232:1247 the second sugar (GlcNAc3NAcA) 1232:1261 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 13 62 theme strain 2461:2466 arg1 pili 2442:2445 the pili 2438:2445 the pili of one mutant strain 2438:2466 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 0 63 theme same 148:151 arg1 pathway 153:159 the same pathway 144:159 the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar 144:222 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 10 64 theme adjacent 1926:1933 arg1 operons 1935:1941 adjacent operons 1926:1941 adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 1926:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 10 64 theme adjacent 1926:1933 arg1 enzymes 1972:1978 the enzymes 1968:1978 the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 1968:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 5 65 from deletions 894:902 arg1 mmp0353 937:943 mmp0353 937:943 mmp0353 937:943 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 5 65 from deletions 894:902 arg1 mmp0351 916:922 mmp0351 916:922 mmp0351 916:922 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 5 65 from deletions 894:902 arg1 mmp0350 907:913 mmp0350 907:913 mmp0350 907:913 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 5 65 from deletions 894:902 arg1 mmp0352 925:931 mmp0352 925:931 mmp0352 925:931 Mutants carrying deletions in mmp0350, mmp0351, mmp0352, or mmp0353 were nonarchaellated and synthesized archaellins modified with a 1-sugar glycan, as estimated from Western blots. 25733616 11 66 theme archaellum 2235:2244 arg1 assembly 2246:2253 archaellum assembly 2235:2253 archaellum assembly 2235:2253 This posttranslational modification of archaellins is important, as it is necessary for archaellum assembly. 25733616 0 67 from archaeon 99:106 arg1 sugars 51:56 the second and third sugars 30:56 the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 30:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 7 68 theme third 1454:1458 arg1 ManNAc3NAmA6Thr 1467:1481 ManNAc3NAmA6Thr 1467:1481 ManNAc3NAmA6Thr 1467:1481 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 7 68 theme third 1454:1458 arg1 sugar 1460:1464 the third sugar 1450:1464 the third sugar (ManNAc3NAmA6Thr) 1450:1482 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 13 69 theme pili 2442:2445 arg1 analysis 2426:2433 Mass spectrometry analysis 2408:2433 Mass spectrometry analysis of the pili of one mutant strain 2408:2466 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 7 70 with archaellins 1309:1319 arg1 glycan 1336:1341 a 2-sugar glycan 1326:1341 a 2-sugar glycan 1326:1341 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 3 71 contain has 606:608 arg2 hexose 624:629 an additional hexose 610:629 an additional hexose attached to GalNAc 610:648 The pilin glycan has an additional hexose attached to GalNAc. 25733616 3 71 contain has 606:608 arg1 glycan 599:604 The pilin glycan 589:604 The pilin glycan 589:604 The pilin glycan has an additional hexose attached to GalNAc. 25733616 10 72 dep IMPORTANCE 1873:1882 arg1 identifies 1894:1903 identifies 1894:1903 identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 1894:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 10 73 theme maripaludis 2134:2144 arg1 archaellins 2105:2115 archaellins 2105:2115 archaellins of Methanococcus maripaludis 2105:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 0 74 theme archaellin 65:74 arg1 Tetrasaccharide 76:90 the archaellin Tetrasaccharide 61:90 the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 61:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 13 75 theme spectrometry 2413:2424 arg1 analysis 2426:2433 Mass spectrometry analysis 2408:2433 Mass spectrometry analysis of the pili of one mutant strain 2408:2466 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 1 76 theme glycoprotein 340:351 arg1 subunits 353:360 glycoprotein subunits 340:360 glycoprotein subunits 340:360 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 0 77 dep archaeon 99:106 arg1 maripaludis 122:132 Methanococcus maripaludis 108:132 the archaeon Methanococcus maripaludis 95:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 4 78 theme N-glycan 860:867 arg1 sugars 869:874 N-glycan sugars 860:874 N-glycan sugars 860:874 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 12 79 attach attached 2377:2384 arg1 sugar 2401:2405 the linking sugar 2389:2405 the linking sugar 2389:2405 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 12 79 attach attached 2377:2384 arg2 hexose 2370:2375 an additional hexose 2356:2375 an additional hexose attached to the linking sugar 2356:2405 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 12 80 theme linking 2393:2399 arg1 sugar 2401:2405 the linking sugar 2389:2405 the linking sugar 2389:2405 Pilins are modified with a different N-glycan consisting of the archaellin tetrasaccharide but with an additional hexose attached to the linking sugar. 25733616 4 81 from involvement 829:839 arg1 biosynthesis 844:855 biosynthesis 844:855 biosynthesis of N-glycan sugars 844:874 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 7 82 theme archaella 1390:1398 arg1 spectroscopy 1365:1376 mass spectroscopy 1360:1376 mass spectroscopy of purified archaella 1360:1398 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 0 83 theme second 34:39 arg1 sugars 51:56 the second and third sugars 30:56 the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 30:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 1 84 theme type 304:307 arg1 pili 312:315 type IV pili 304:315 type IV pili 304:315 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 1 84 theme type 304:307 arg1 appendages 278:287 two surface appendages 266:287 two surface appendages 266:287 UNLABELLED Methanococcus maripaludis has two surface appendages, archaella and type IV pili, which are composed of glycoprotein subunits. 25733616 2 85 theme 5S 512:513 arg1 -2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose 515:586 (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose 511:586 (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose 511:586 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 2 86 link N-linked 396:403 arg1 tetrasaccharide 405:419 an N-linked tetrasaccharide 393:419 an N-linked tetrasaccharide 393:419 Archaellins are modified with an N-linked tetrasaccharide with the structure Sug-1,4-β-ManNAc3NAmA6Thr-1,4-β-GlcNAc3NAcA-1,3-β-GalNAc, where Sug is (5S)-2-acetamido-2,4-dideoxy-5-O-methyl-α-L-erythro-hexos-5-ulo-1,5-pyranose. 25733616 6 87 dep mmp0353 1187:1193 arg1 to 1184:1185 to 1184:1185 to 1184:1185 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 14 88 theme archaeal 2588:2595 arg1 gene 2597:2600 an archaeal gene 2585:2600 an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway 2585:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 0 89 theme third 45:49 arg1 sugars 51:56 the second and third sugars 30:56 the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis 30:132 Evidence that biosynthesis of the second and third sugars of the archaellin Tetrasaccharide in the archaeon Methanococcus maripaludis occurs by the same pathway used by Pseudomonas aeruginosa to make a di-N-acetylated sugar. 25733616 4 90 dep operons 721:727 arg1 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 90 dep operons 721:727 arg1 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 90 dep operons 721:727 arg1 operons 721:727 two adjacent, divergently transcribed operons 683:727 two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) 683:765 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 7 91 theme mass 1360:1363 arg1 spectroscopy 1365:1376 mass spectroscopy 1360:1376 mass spectroscopy of purified archaella 1360:1398 The Δmmp0357 mutant was archaellated and had archaellins with a 2-sugar glycan, as confirmed by mass spectroscopy of purified archaella, indicating a role for MMP0357 in biosynthesis of the third sugar (ManNAc3NAmA6Thr). 25733616 8 92 theme UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic 1677:1720 arg1 sugar 1751:1755 an O5-specific antigen sugar 1728:1755 an O5-specific antigen sugar 1728:1755 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 8 92 theme UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic 1677:1720 arg1 acid 1722:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 3 93 theme pilin 593:597 arg1 glycan 599:604 The pilin glycan 589:604 The pilin glycan 589:604 The pilin glycan has an additional hexose attached to GalNAc. 25733616 6 94 with glycan 1140:1145 arg1 GalNAc 1157:1162 only GalNAc 1152:1162 only GalNAc 1152:1162 Mass spectroscopy analysis of pili purified from the Δmmp0352 strain confirmed a glycan with only GalNAc, suggesting mmp0350 to mmp0353 were all involved in biosynthesis of the second sugar (GlcNAc3NAcA). 25733616 13 95 theme different 2499:2507 arg1 glycan 2509:2514 this different glycan 2494:2514 this different glycan 2494:2514 Mass spectrometry analysis of the pili of one mutant strain provided insight into how this different glycan might ultimately be assembled. 25733616 4 96 theme adjacent 687:694 arg1 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 mmp0359-mmp0355 750:764 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 96 theme adjacent 687:694 arg1 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 mmp0350-mmp0354 730:744 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 4 96 theme adjacent 687:694 arg1 operons 721:727 two adjacent, divergently transcribed operons 683:727 two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) 683:765 In this study, genes located in two adjacent, divergently transcribed operons (mmp0350-mmp0354 and mmp0359-mmp0355) were targeted for study based on annotations suggesting their involvement in biosynthesis of N-glycan sugars. 25733616 10 97 theme entire 1998:2003 arg1 pathway 2005:2011 the entire pathway 1994:2011 the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 1994:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 14 98 theme rare 2569:2572 arg1 example 2574:2580 a rare example 2567:2580 a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway 2567:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 8 99 dep Pseudomonas 1594:1604 arg1 aeruginosa 1606:1615 aeruginosa 1606:1615 aeruginosa 1606:1615 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 14 100 theme complex 2647:2653 arg1 pathway 2674:2680 a complex sugar biosynthesis pathway 2645:2680 a complex sugar biosynthesis pathway 2645:2680 This study includes a rare example of an archaeal gene functionally replacing a bacterial gene in a complex sugar biosynthesis pathway. 25733616 8 101 theme O5-specific 1731:1741 arg1 sugar 1751:1755 an O5-specific antigen sugar 1728:1755 an O5-specific antigen sugar 1728:1755 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 8 101 theme O5-specific 1731:1741 arg1 acid 1722:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid 1677:1725 M. maripaludis mmp0350, mmp0351, mmp0352, mmp0353, and mmp0357 are proposed to be functionally equivalent to Pseudomonas aeruginosa wbpABEDI, involved in converting UDP-N-acetylglucosamine to UDP-2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, an O5-specific antigen sugar. 25733616 10 102 theme second 2035:2040 arg1 sugars 2052:2057 the second and third sugars 2031:2057 the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis 2031:2144 IMPORTANCE This work identifies a series of genes in adjacent operons that are shown to encode the enzymes that complete the entire pathway for generation of the second and third sugars of the N-linked tetrasaccharide that modifies archaellins of Methanococcus maripaludis. 25733616 3 103 theme additional 613:622 arg1 hexose 624:629 an additional hexose 610:629 an additional hexose attached to GalNAc 610:648 The pilin glycan has an additional hexose attached to GalNAc. 29194986 7 0 theme plasmon 1259:1265 arg1 resonance 1267:1275 surface plasmon resonance 1251:1275 surface plasmon resonance (SPR) 1251:1281 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 0 theme plasmon 1259:1265 arg1 SPR 1278:1280 SPR 1278:1280 SPR 1278:1280 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 1 theme Fcγ 1185:1187 arg1 IIIa 1210:1213 IIIa 1210:1213 IIIa 1210:1213 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 1 theme Fcγ 1185:1187 arg1 FcγRI 1200:1204 FcγRI 1200:1204 FcγRI 1200:1204 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 1 theme Fcγ 1185:1187 arg1 receptors 1189:1197 two human Fcγ receptors 1175:1197 two human Fcγ receptors (FcγRI and IIIa) 1175:1214 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 1 2 theme cell 163:166 arg1 biofactories 168:179 other cell biofactories 157:179 other cell biofactories commonly used to produce monoclonal antibodies 157:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 4 3 theme recombinant 643:653 arg1 antibody 655:662 this recombinant antibody 638:662 this recombinant antibody 638:662 A biochemical characterization of this recombinant antibody demonstrated that the Asn-297 is N-glycosylated by oligomannosides. 29194986 8 4 with FcyRIIIa 1401:1408 arg1 affinity 1430:1437 3-times higher affinity 1415:1437 3-times higher affinity compared to a control human IgG1 1415:1470 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 5 5 theme immune 739:744 arg1 system 746:751 the immune system 735:751 the immune system 735:751 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 3 6 theme tricornutum 591:601 arg1 medium 556:561 the culture medium 544:561 the culture medium of the diatom Phaeodactylum tricornutum 544:601 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 2 7 theme low 358:360 arg1 costs 373:377 their phototrophic lifestyle allowing low production costs 320:377 their phototrophic lifestyle allowing low production costs 320:377 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 7 theme low 358:360 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 7 8 theme human 1179:1183 arg1 IIIa 1210:1213 IIIa 1210:1213 IIIa 1210:1213 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 8 theme human 1179:1183 arg1 FcγRI 1200:1204 FcγRI 1200:1204 FcγRI 1200:1204 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 8 theme human 1179:1183 arg1 receptors 1189:1197 two human Fcγ receptors 1175:1197 two human Fcγ receptors (FcγRI and IIIa) 1175:1214 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 3 9 theme culture 548:554 arg1 medium 556:561 the culture medium 544:561 the culture medium of the diatom Phaeodactylum tricornutum 544:601 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 4 10 theme antibody 655:662 arg1 characterization 618:633 A biochemical characterization 604:633 A biochemical characterization of this recombinant antibody 604:662 A biochemical characterization of this recombinant antibody demonstrated that the Asn-297 is N-glycosylated by oligomannosides. 29194986 5 11 theme Fc 822:823 arg1 part 825:828 their Fc part 816:828 their Fc part 816:828 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 8 12 theme alga-made 1324:1332 arg1 antibody 1334:1341 the alga-made antibody 1320:1341 the alga-made antibody 1320:1341 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 8 12 theme alga-made 1324:1332 arg1 able 1346:1349 able 1346:1349 able 1346:1349 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 4 13 gly N-glycosylated 697:710 arg1 N-glycosylated 697:710 N-glycosylated 697:710 N-glycosylated by oligomannosides 697:729 A biochemical characterization of this recombinant antibody demonstrated that the Asn-297 is N-glycosylated by oligomannosides. 29194986 4 13 gly N-glycosylated 697:710 arg1 Asn-297 686:692 the Asn-297 682:692 the Asn-297 682:692 A biochemical characterization of this recombinant antibody demonstrated that the Asn-297 is N-glycosylated by oligomannosides. 29194986 3 14 theme diatom 570:575 arg1 tricornutum 591:601 the diatom Phaeodactylum tricornutum 566:601 the diatom Phaeodactylum tricornutum 566:601 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 3 15 theme Hepatitis 488:496 arg1 antigen 508:514 Hepatitis B surface antigen 488:514 Hepatitis B surface antigen 488:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 3 16 theme recombinant 446:456 arg1 antibody 462:469 a fully assembled recombinant IgG antibody 428:469 a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen 428:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 5 17 theme Fcγ 853:855 arg1 FcγR 868:871 FcγR 868:871 FcγR 868:871 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 5 17 theme Fcγ 853:855 arg1 receptors 857:865 Fcγ receptors 853:865 Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes 853:940 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 2 18 theme growth 304:309 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 3 19 theme B 498:498 arg1 antigen 508:514 Hepatitis B surface antigen 488:514 Hepatitis B surface antigen 488:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 0 20 theme Anti-Hepatitis 10:23 arg1 Antibody 27:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody Binds to Human Fcγ Receptors. 29194986 1 21 theme unicellular 81:91 arg1 Microalgae 66:75 Microalgae 66:75 Microalgae 66:75 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 1 21 theme unicellular 81:91 arg1 organisms 104:112 unicellular eukaryotic organisms 81:112 unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies 81:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 7 22 theme anti-hepatitis 1112:1125 arg1 IgG 1141:1143 the anti-hepatitis B recombinant IgG 1108:1143 the anti-hepatitis B recombinant IgG produced in P. tricornutum 1108:1170 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 0 23 theme Alga-Made 0:8 arg1 Antibody 27:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody Binds to Human Fcγ Receptors. 29194986 4 24 theme biochemical 606:616 arg1 characterization 618:633 A biochemical characterization 604:633 A biochemical characterization of this recombinant antibody 604:662 A biochemical characterization of this recombinant antibody demonstrated that the Asn-297 is N-glycosylated by oligomannosides. 29194986 5 25 theme opsonized 923:931 arg1 microbes 933:940 opsonized microbes 923:940 opsonized microbes 923:940 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 2 26 theme allowing 349:356 arg1 costs 373:377 their phototrophic lifestyle allowing low production costs 320:377 their phototrophic lifestyle allowing low production costs 320:377 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 26 theme allowing 349:356 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 5 27 theme effector 779:786 arg1 molecules 788:796 effector molecules 779:796 effector molecules 779:796 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 2 28 theme lifestyle 339:347 arg1 costs 373:377 their phototrophic lifestyle allowing low production costs 320:377 their phototrophic lifestyle allowing low production costs 320:377 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 28 theme lifestyle 339:347 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 8 29 theme higher 1423:1428 arg1 affinity 1430:1437 3-times higher affinity 1415:1437 3-times higher affinity compared to a control human IgG1 1415:1470 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 1 30 theme monoclonal 206:215 arg1 antibodies 217:226 monoclonal antibodies 206:226 monoclonal antibodies 206:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 1 31 theme eukaryotic 93:102 arg1 Microalgae 66:75 Microalgae 66:75 Microalgae 66:75 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 1 31 theme eukaryotic 93:102 arg1 organisms 104:112 unicellular eukaryotic organisms 81:112 unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies 81:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 2 32 theme phototrophic 326:337 arg1 costs 373:377 their phototrophic lifestyle allowing low production costs 320:377 their phototrophic lifestyle allowing low production costs 320:377 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 32 theme phototrophic 326:337 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 3 33 theme IgG 458:460 arg1 antibody 462:469 a fully assembled recombinant IgG antibody 428:469 a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen 428:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 7 34 theme B 1127:1127 arg1 IgG 1141:1143 the anti-hepatitis B recombinant IgG 1108:1143 the anti-hepatitis B recombinant IgG produced in P. tricornutum 1108:1170 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 35 theme cellular 1224:1231 arg1 assay 1241:1245 a cellular binding assay 1222:1245 a cellular binding assay 1222:1245 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 6 36 attach present 1019:1025 arg1 Asn-297 1034:1040 the Asn-297 1030:1040 the Asn-297 1030:1040 Interactions between IgG and FcγR are influenced by the N-glycan structures present on the Asn-297. 29194986 6 36 attach present 1019:1025 arg2 structures 1008:1017 the N-glycan structures 995:1017 the N-glycan structures present on the Asn-297 995:1040 Interactions between IgG and FcγR are influenced by the N-glycan structures present on the Asn-297. 29194986 3 37 theme surface 500:506 arg1 antigen 508:514 Hepatitis B surface antigen 488:514 Hepatitis B surface antigen 488:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 5 38 theme receptors 857:865 arg1 part 825:828 their Fc part 816:828 their Fc part 816:828 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 5 38 theme receptors 857:865 arg1 recognition 838:848 the recognition 834:848 the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes 834:940 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 7 39 theme recombinant 1129:1139 arg1 IgG 1141:1143 the anti-hepatitis B recombinant IgG 1108:1143 the anti-hepatitis B recombinant IgG produced in P. tricornutum 1108:1170 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 2 40 theme biotechnological 256:271 arg1 advantages 273:282 several biotechnological advantages 248:282 several biotechnological advantages such as their rapid growth rate 248:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 40 theme biotechnological 256:271 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 0 41 theme Fcγ 51:53 arg1 Receptors 55:63 Human Fcγ Receptors 45:63 Human Fcγ Receptors 45:63 Alga-Made Anti-Hepatitis B Antibody Binds to Human Fcγ Receptors. 29194986 6 42 theme present 1019:1025 arg1 structures 1008:1017 the N-glycan structures 995:1017 the N-glycan structures present on the Asn-297 995:1040 Interactions between IgG and FcγR are influenced by the N-glycan structures present on the Asn-297. 29194986 2 43 theme several 248:254 arg1 advantages 273:282 several biotechnological advantages 248:282 several biotechnological advantages such as their rapid growth rate 248:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 43 theme several 248:254 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 7 44 theme binding 1088:1094 arg1 capacity 1096:1103 the binding capacity 1084:1103 the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) 1084:1214 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 2 45 theme protein 382:388 arg1 expression 390:399 protein expression 382:399 protein expression 382:399 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 0 46 theme Human 45:49 arg1 Receptors 55:63 Human Fcγ Receptors 45:63 Human Fcγ Receptors 45:63 Alga-Made Anti-Hepatitis B Antibody Binds to Human Fcγ Receptors. 29194986 7 47 theme IgG 1141:1143 arg1 capacity 1096:1103 the binding capacity 1084:1103 the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) 1084:1214 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 8 48 theme control 1453:1459 arg1 IgG1 1467:1470 a control human IgG1 1451:1470 a control human IgG1 1451:1470 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 6 49 theme N-glycan 999:1006 arg1 structures 1008:1017 the N-glycan structures 995:1017 the N-glycan structures present on the Asn-297 995:1040 Interactions between IgG and FcγR are influenced by the N-glycan structures present on the Asn-297. 29194986 2 50 theme rapid 298:302 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 1 51 theme emerging 133:140 arg1 alternative 142:152 an emerging alternative 130:152 an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies 130:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 8 52 theme reduced 1372:1378 arg1 affinity 1380:1387 a reduced affinity 1370:1387 a reduced affinity 1370:1387 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 3 53 theme Phaeodactylum 577:589 arg1 tricornutum 591:601 the diatom Phaeodactylum tricornutum 566:601 the diatom Phaeodactylum tricornutum 566:601 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 2 54 theme production 362:371 arg1 costs 373:377 their phototrophic lifestyle allowing low production costs 320:377 their phototrophic lifestyle allowing low production costs 320:377 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 2 54 theme production 362:371 arg1 rate 311:314 their rapid growth rate 292:314 their rapid growth rate 292:314 Microalgae display several biotechnological advantages such as their rapid growth rate and their phototrophic lifestyle allowing low production costs as protein expression is solar-fueled. 29194986 8 55 theme human 1461:1465 arg1 IgG1 1467:1470 a control human IgG1 1451:1470 a control human IgG1 1451:1470 This allowed us to demonstrate that the alga-made antibody is able to bind FcγRI with a reduced affinity and engages FcyRIIIa with 3-times higher affinity compared to a control human IgG1. 29194986 7 56 dep receptors 1189:1197 arg1 IIIa 1210:1213 IIIa 1210:1213 IIIa 1210:1213 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 56 dep receptors 1189:1197 arg1 FcγRI 1200:1204 FcγRI 1200:1204 FcγRI 1200:1204 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 56 dep receptors 1189:1197 arg1 receptors 1189:1197 two human Fcγ receptors 1175:1197 two human Fcγ receptors (FcγRI and IIIa) 1175:1214 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 5 57 theme microbes 933:940 arg1 phagocytosis 907:918 phagocytosis 907:918 phagocytosis of opsonized microbes 907:940 In the immune system, antibodies interact with effector molecules and cells through their Fc part and the recognition of Fcγ receptors (FcγR) which are important for inducing phagocytosis of opsonized microbes. 29194986 3 58 theme assembled 436:444 arg1 antibody 462:469 a fully assembled recombinant IgG antibody 428:469 a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen 428:514 Recently, a fully assembled recombinant IgG antibody directed against Hepatitis B surface antigen is produced and secreted in the culture medium of the diatom Phaeodactylum tricornutum. 29194986 7 59 theme binding 1233:1239 arg1 assay 1241:1245 a cellular binding assay 1222:1245 a cellular binding assay 1222:1245 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 0 60 theme B 25:25 arg1 Antibody 27:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody 0:34 Alga-Made Anti-Hepatitis B Antibody Binds to Human Fcγ Receptors. 29194986 1 61 theme other 157:161 arg1 biofactories 168:179 other cell biofactories 157:179 other cell biofactories commonly used to produce monoclonal antibodies 157:226 Microalgae are unicellular eukaryotic organisms which represent an emerging alternative to other cell biofactories commonly used to produce monoclonal antibodies. 29194986 7 62 theme surface 1251:1257 arg1 resonance 1267:1275 surface plasmon resonance 1251:1275 surface plasmon resonance (SPR) 1251:1281 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 29194986 7 62 theme surface 1251:1257 arg1 SPR 1278:1280 SPR 1278:1280 SPR 1278:1280 In this study, the authors characterized the binding capacity of the anti-hepatitis B recombinant IgG produced in P. tricornutum to two human Fcγ receptors (FcγRI and IIIa) using a cellular binding assay and surface plasmon resonance (SPR). 28939751 0 0 from development 8:18 arg1 regulation 29:38 Notch regulation 23:38 Notch regulation 23:38 A sweet development in Notch regulation. 28939751 3 1 theme glycosylation 447:459 arg1 types 438:442 different types 428:442 different types of glycosylation 428:459 This work shows how different types of glycosylation can distinctly influence protein stability and structure. 28939751 1 2 theme transmembrane 45:57 arg1 Notch 77:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 2 theme transmembrane 45:57 arg1 crucial 93:99 crucial 93:99 crucial 93:99 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 3 theme signaling 59:67 arg1 Notch 77:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 3 theme signaling 59:67 arg1 crucial 93:99 crucial 93:99 crucial 93:99 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 3 4 theme different 428:436 arg1 types 438:442 different types 428:442 different types of glycosylation 428:459 This work shows how different types of glycosylation can distinctly influence protein stability and structure. 28939751 1 5 contain has 136:138 arg1 crucial 93:99 crucial 93:99 crucial 93:99 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 5 contain has 136:138 arg1 Notch 77:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 5 contain has 136:138 arg2 domains 161:167 36 extracellular EGF domains 140:167 36 extracellular EGF domains that are glycosylated in variable and complex ways 140:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 6 gly glycosylated 178:189 arg1 domains 161:167 36 extracellular EGF domains 140:167 36 extracellular EGF domains that are glycosylated in variable and complex ways 140:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 6 gly glycosylated 178:189 arg1 ways 215:218 variable and complex ways 194:218 variable and complex ways 194:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 6 gly glycosylated 178:189 arg2 domains 161:167 36 extracellular EGF domains 140:167 36 extracellular EGF domains that are glycosylated in variable and complex ways 140:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 7 theme protein 69:75 arg1 Notch 77:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch 41:81 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 7 theme protein 69:75 arg1 crucial 93:99 crucial 93:99 crucial 93:99 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 8 theme extracellular 143:155 arg1 domains 161:167 36 extracellular EGF domains 140:167 36 extracellular EGF domains that are glycosylated in variable and complex ways 140:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 9 theme EGF 157:159 arg1 domains 161:167 36 extracellular EGF domains 140:167 36 extracellular EGF domains that are glycosylated in variable and complex ways 140:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 0 10 theme sweet 2:6 arg1 development 8:18 A sweet development 0:18 A sweet development in Notch regulation 0:38 A sweet development in Notch regulation. 28939751 2 11 theme protein 392:398 arg1 groove 400:405 a protein groove 390:405 a protein groove 390:405 A new study shows that O-fucose and O-glucose stabilize the repeats but that extension of glucose by xylose weakens stability, explained by the binding of the glycan to a protein groove. 28939751 2 12 theme glycan 380:385 arg1 binding 365:371 the binding 361:371 the binding of the glycan to a protein groove 361:405 A new study shows that O-fucose and O-glucose stabilize the repeats but that extension of glucose by xylose weakens stability, explained by the binding of the glycan to a protein groove. 28939751 3 13 theme protein 486:492 arg1 stability 494:502 protein stability 486:502 protein stability 486:502 This work shows how different types of glycosylation can distinctly influence protein stability and structure. 28939751 0 14 theme Notch 23:27 arg1 regulation 29:38 Notch regulation 23:38 Notch regulation 23:38 A sweet development in Notch regulation. 28939751 1 15 theme variable 194:201 arg1 ways 215:218 variable and complex ways 194:218 variable and complex ways 194:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 2 16 theme glucose 311:317 arg1 extension 298:306 extension 298:306 extension of glucose by xylose 298:327 A new study shows that O-fucose and O-glucose stabilize the repeats but that extension of glucose by xylose weakens stability, explained by the binding of the glycan to a protein groove. 28939751 1 17 theme embryonic 105:113 arg1 decisions 125:133 embryonic cell fate decisions 105:133 embryonic cell fate decisions 105:133 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 18 theme cell 115:118 arg1 decisions 125:133 embryonic cell fate decisions 105:133 embryonic cell fate decisions 105:133 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 19 theme fate 120:123 arg1 decisions 125:133 embryonic cell fate decisions 105:133 embryonic cell fate decisions 105:133 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 1 20 theme complex 207:213 arg1 ways 215:218 variable and complex ways 194:218 variable and complex ways 194:218 The transmembrane signaling protein Notch, which is crucial for embryonic cell fate decisions, has 36 extracellular EGF domains that are glycosylated in variable and complex ways. 28939751 2 21 theme new 223:225 arg1 study 227:231 A new study 221:231 A new study 221:231 A new study shows that O-fucose and O-glucose stabilize the repeats but that extension of glucose by xylose weakens stability, explained by the binding of the glycan to a protein groove. 25296196 4 0 from changes 643:649 arg1 ones 677:680 O-glycans and moderate ones 654:680 O-glycans and moderate ones 654:680 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 4 0 from changes 643:649 arg1 pools 694:698 N-glycan pools 685:698 N-glycan pools 685:698 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 1 1 theme bus-4 112:116 arg1 mutants 138:144 bus-4 glycosyltransferase mutants 112:144 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 7 2 from increase 1141:1148 arg1 expression 1178:1187 GDP-Man 4,6 dehydratease expression 1153:1187 GDP-Man 4,6 dehydratease expression 1153:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 9 3 theme combined 1330:1337 arg1 changes 1339:1345 The combined changes 1326:1345 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin 1326:1442 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 4 theme normal 1519:1524 arg1 cues 1551:1554 normal glycoconjugate dependent cues 1519:1554 normal glycoconjugate dependent cues leading to barriers to these bacterial infections 1519:1604 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 6 5 theme fucosyl 923:929 arg1 O-glycans 889:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 5 theme fucosyl 923:929 arg1 forms 931:935 neutral fucosyl forms 915:935 neutral fucosyl forms 915:935 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 6 theme neutral 915:921 arg1 O-glycans 889:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 6 theme neutral 915:921 arg1 forms 931:935 neutral fucosyl forms 915:935 neutral fucosyl forms 915:935 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 9 7 theme mucin 1438:1442 arg1 distribution 1378:1389 cell surface glycoconjugate distribution 1350:1389 cell surface glycoconjugate distribution 1350:1389 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 7 theme mucin 1438:1442 arg1 abundance 1402:1410 increased abundance 1392:1410 increased abundance 1392:1410 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 7 theme mucin 1438:1442 arg1 properties 1424:1433 altered properties 1416:1433 altered properties 1416:1433 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 8 theme above 1484:1488 arg1 pathogens 1490:1498 the above pathogens 1480:1498 the above pathogens 1480:1498 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 7 9 theme Quantitative 998:1009 arg1 analysis 1022:1029 Quantitative expression analysis 998:1029 Quantitative expression analysis 998:1029 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 6 10 theme fucosyl 979:985 arg1 N-glycans 987:995 fucosyl N-glycans 979:995 fucosyl N-glycans 979:995 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 11 theme Ce 900:901 arg1 O-glycans 889:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 11 theme Ce 900:901 arg1 core-II 903:909 Ce core-II 900:909 Ce core-II 900:909 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 7 12 theme 4,6 1161:1163 arg1 expression 1178:1187 GDP-Man 4,6 dehydratease expression 1153:1187 GDP-Man 4,6 dehydratease expression 1153:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 3 13 theme structural 567:576 arg1 detail 578:583 fine structural detail 562:583 fine structural detail for the bus-4 N- and O-glycan pools 562:619 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 7 14 dep revealed 1117:1124 arg1 increase 1141:1148 a dramatic increase 1130:1148 a dramatic increase in GDP-Man 4,6 dehydratease expression 1130:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 5 15 theme mucin 765:769 arg1 glycans 741:747 Ce core-I glycans 731:747 Ce core-I glycans 731:747 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 5 15 theme mucin 765:769 arg1 glycan 771:776 the nematode's mucin glycan 750:776 the nematode's mucin glycan equivalent 750:787 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 8 16 theme binding 1219:1225 arg1 studies 1227:1233 detailed lectin binding studies 1203:1233 detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine 1203:1323 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 1 17 theme Caenorabditis 90:102 arg1 mutants 138:144 bus-4 glycosyltransferase mutants 112:144 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 4 18 theme N-glycan 685:692 arg1 pools 694:698 N-glycan pools 685:698 N-glycan pools 685:698 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 7 19 theme dehydratease 1165:1176 arg1 expression 1178:1187 GDP-Man 4,6 dehydratease expression 1153:1187 GDP-Man 4,6 dehydratease expression 1153:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 3 20 theme fine 562:565 arg1 detail 578:583 fine structural detail 562:583 fine structural detail for the bus-4 N- and O-glycan pools 562:619 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 9 21 from changes 1339:1345 arg1 distribution 1378:1389 cell surface glycoconjugate distribution 1350:1389 cell surface glycoconjugate distribution 1350:1389 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 21 from changes 1339:1345 arg1 abundance 1402:1410 increased abundance 1392:1410 increased abundance 1392:1410 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 9 21 from changes 1339:1345 arg1 properties 1424:1433 altered properties 1416:1433 altered properties 1416:1433 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 0 22 theme Caenorhabditis 0:13 arg1 mutants 58:64 bacterial pathogen resistant bus-4 mutants 23:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 3 23 theme O-glycan 606:613 arg1 pools 615:619 the bus-4 N- and O-glycan pools 589:619 the bus-4 N- and O-glycan pools 589:619 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 1 24 theme Yersinia 225:232 arg1 pseudotuberculosis 234:251 Yersinia pseudotuberculosis 225:251 Yersinia pseudotuberculosis 225:251 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 0 25 theme pathogen 33:40 arg1 mutants 58:64 bacterial pathogen resistant bus-4 mutants 23:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 5 26 theme Ce 731:732 arg1 glycans 741:747 Ce core-I glycans 731:747 Ce core-I glycans 731:747 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 5 26 theme Ce 731:732 arg1 glycan 771:776 the nematode's mucin glycan 750:776 the nematode's mucin glycan equivalent 750:787 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 8 27 theme detailed 1203:1210 arg1 studies 1227:1233 detailed lectin binding studies 1203:1233 detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine 1203:1323 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 1 28 theme glycosyltransferase 118:136 arg1 mutants 138:144 bus-4 glycosyltransferase mutants 112:144 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 9 29 theme increased 1392:1400 arg1 abundance 1402:1410 increased abundance 1392:1410 increased abundance 1392:1410 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 3 30 theme MS 516:517 arg1 experiments 540:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 2 31 theme pathogen 472:479 arg1 resistance 481:490 pathogen resistance 472:490 pathogen resistance 472:490 Our objective in this study was to define the glycosylation changes leading to this phenotype to better understand how these changes lead to pathogen resistance. 25296196 9 32 theme cell 1350:1353 arg1 distribution 1378:1389 cell surface glycoconjugate distribution 1350:1389 cell surface glycoconjugate distribution 1350:1389 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 5 33 theme core-I 734:739 arg1 glycans 741:747 Ce core-I glycans 731:747 Ce core-I glycans 731:747 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 5 33 theme core-I 734:739 arg1 glycan 771:776 the nematode's mucin glycan 750:776 the nematode's mucin glycan equivalent 750:787 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 6 34 theme fucosyl 881:887 arg1 O-glycans 889:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans 877:897 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 34 theme fucosyl 881:887 arg1 core-II 903:909 Ce core-II 900:909 Ce core-II 900:909 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 34 theme fucosyl 881:887 arg1 forms 931:935 neutral fucosyl forms 915:935 neutral fucosyl forms 915:935 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 3 35 theme N- 599:600 arg1 pools 615:619 the bus-4 N- and O-glycan pools 589:619 the bus-4 N- and O-glycan pools 589:619 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 9 36 theme dependent 1541:1549 arg1 cues 1551:1554 normal glycoconjugate dependent cues 1519:1554 normal glycoconjugate dependent cues leading to barriers to these bacterial infections 1519:1604 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 0 37 theme bus-4 52:56 arg1 mutants 58:64 bacterial pathogen resistant bus-4 mutants 23:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 3 38 theme GC/MS 534:538 arg1 experiments 540:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 0 39 theme resistant 42:50 arg1 mutants 58:64 bacterial pathogen resistant bus-4 mutants 23:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 9 40 theme glycoconjugate 1363:1376 arg1 distribution 1378:1389 cell surface glycoconjugate distribution 1350:1389 cell surface glycoconjugate distribution 1350:1389 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 8 41 theme lectin 1212:1217 arg1 studies 1227:1233 detailed lectin binding studies 1203:1233 detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine 1203:1323 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 0 42 dep mutants 58:64 arg1 elegans 15:21 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 5 43 theme terminal 853:860 arg1 substitutions 862:874 terminal substitutions 853:874 terminal substitutions 853:874 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 1 44 theme altered 262:268 arg1 susceptibility 270:283 altered susceptibility 262:283 altered susceptibility to two Leucobacter species Verde1 and Verde2 262:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 5 45 theme equivalent 778:787 arg1 glycans 741:747 Ce core-I glycans 731:747 Ce core-I glycans 731:747 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 5 45 theme equivalent 778:787 arg1 glycan 771:776 the nematode's mucin glycan 750:776 the nematode's mucin glycan equivalent 750:787 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 8 46 theme cuticle 1295:1301 arg1 surface 1303:1309 cuticle surface 1295:1309 cuticle surface 1295:1309 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 7 47 theme dramatic 1132:1139 arg1 increase 1141:1148 a dramatic increase 1130:1148 a dramatic increase in GDP-Man 4,6 dehydratease expression 1130:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 7 48 theme expression 1011:1020 arg1 analysis 1022:1029 Quantitative expression analysis 998:1029 Quantitative expression analysis 998:1029 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 2 49 theme glycosylation 377:389 arg1 changes 391:397 the glycosylation changes 373:397 the glycosylation changes leading to this phenotype to better understand how these changes lead to pathogen resistance 373:490 Our objective in this study was to define the glycosylation changes leading to this phenotype to better understand how these changes lead to pathogen resistance. 25296196 7 50 theme GDP-Man 1153:1159 arg1 expression 1178:1187 GDP-Man 4,6 dehydratease expression 1153:1187 GDP-Man 4,6 dehydratease expression 1153:1187 Quantitative expression analysis revealed that two mucins, let-653 and osm-8, were upregulated nearly 40 fold and also revealed was a dramatic increase in GDP-Man 4,6 dehydratease expression. 25296196 1 51 contain have 257:260 arg2 susceptibility 270:283 altered susceptibility 262:283 altered susceptibility to two Leucobacter species Verde1 and Verde2 262:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 51 contain have 257:260 arg1 mutants 138:144 bus-4 glycosyltransferase mutants 112:144 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 9 52 theme surface 1355:1361 arg1 distribution 1378:1389 cell surface glycoconjugate distribution 1350:1389 cell surface glycoconjugate distribution 1350:1389 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 5 53 from shifts 843:848 arg1 substitutions 862:874 terminal substitutions 853:874 terminal substitutions 853:874 Ce core-I glycans, the nematode's mucin glycan equivalent, were doubled in abundance, halved in charge and bore shifts in terminal substitutions. 25296196 3 54 theme tandem 520:525 arg1 MS 527:528 tandem MS 520:528 tandem MS 520:528 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 8 55 from changes 1247:1253 arg1 surface 1303:1309 cuticle surface 1295:1309 cuticle surface 1295:1309 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 8 55 from changes 1247:1253 arg1 glycoconjugates 1258:1272 glycoconjugates 1258:1272 glycoconjugates 1258:1272 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 8 55 from changes 1247:1253 arg1 intestine 1315:1323 intestine 1315:1323 intestine 1315:1323 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 8 55 from changes 1247:1253 arg1 coat 1289:1292 the surface coat 1277:1292 the surface coat 1277:1292 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 8 56 theme surface 1281:1287 arg1 coat 1289:1292 the surface coat 1277:1292 the surface coat 1277:1292 We performed detailed lectin binding studies that showed changes in glycoconjugates in the surface coat, cuticle surface and intestine. 25296196 9 57 theme bacterial 1585:1593 arg1 infections 1595:1604 these bacterial infections 1579:1604 these bacterial infections 1579:1604 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 1 58 dep species 304:310 arg1 Verde1 312:317 Verde1 312:317 Verde1 312:317 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 58 dep species 304:310 arg1 species 304:310 two Leucobacter species 288:310 two Leucobacter species Verde1 and Verde2 288:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 58 dep species 304:310 arg1 Verde2 323:328 Verde2 323:328 Verde2 323:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 9 59 theme altered 1416:1422 arg1 properties 1424:1433 altered properties 1416:1433 altered properties 1416:1433 The combined changes in cell surface glycoconjugate distribution, increased abundance and altered properties of mucin provide an environment where likely the above pathogens are not exposed to normal glycoconjugate dependent cues leading to barriers to these bacterial infections. 25296196 3 60 theme MS 527:528 arg1 experiments 540:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 MALDI-TOF MS, tandem MS and GC/MS experiments 506:550 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 4 61 theme dramatic 634:641 arg1 changes 643:649 dramatic changes 634:649 dramatic changes in O-glycans and moderate ones in N-glycan pools 634:698 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 3 62 theme MALDI-TOF 506:514 arg1 MS 516:517 MALDI-TOF MS 506:517 MALDI-TOF MS 506:517 We performed MALDI-TOF MS, tandem MS and GC/MS experiments to reveal fine structural detail for the bus-4 N- and O-glycan pools. 25296196 4 63 theme moderate 668:675 arg1 ones 677:680 O-glycans and moderate ones 654:680 O-glycans and moderate ones 654:680 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 4 64 theme parent 716:721 arg1 strain 723:728 the parent strain 712:728 the parent strain 712:728 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 2 65 from objective 335:343 arg1 study 353:357 this study 348:357 this study 348:357 Our objective in this study was to define the glycosylation changes leading to this phenotype to better understand how these changes lead to pathogen resistance. 25296196 4 66 theme O-glycans 654:662 arg1 ones 677:680 O-glycans and moderate ones 654:680 O-glycans and moderate ones 654:680 We observed dramatic changes in O-glycans and moderate ones in N-glycan pools compared to the parent strain. 25296196 0 67 theme altered 74:80 arg1 mucins 82:87 altered mucins 74:87 altered mucins 74:87 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 6 68 dep N-glycans 987:995 arg1 as 971:972 as 971:972 as 971:972 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 6 68 dep N-glycans 987:995 arg1 were 974:977 were 974:977 were 974:977 The fucosyl O-glycans, Ce core-II and neutral fucosyl forms, were also increased in abundance as were fucosyl N-glycans. 25296196 0 69 theme bacterial 23:31 arg1 mutants 58:64 bacterial pathogen resistant bus-4 mutants 23:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants 0:64 Caenorhabditis elegans bacterial pathogen resistant bus-4 mutants produce altered mucins. 25296196 1 70 dep mutants 138:144 arg1 elegans 104:110 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants 90:144 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 71 theme Leucobacter 292:302 arg1 Verde1 312:317 Verde1 312:317 Verde1 312:317 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 71 theme Leucobacter 292:302 arg1 species 304:310 two Leucobacter species 288:310 two Leucobacter species Verde1 and Verde2 288:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 25296196 1 71 theme Leucobacter 292:302 arg1 Verde2 323:328 Verde2 323:328 Verde2 323:328 Caenorabditis elegans bus-4 glycosyltransferase mutants are resistant to infection by Microbacterium nematophilum, Yersinia pestis and Yersinia pseudotuberculosis and have altered susceptibility to two Leucobacter species Verde1 and Verde2. 28035807 7 0 from level 1380:1384 arg1 samples 1356:1362 complex biological samples 1337:1362 complex biological samples at the proteome level 1337:1384 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 3 1 theme decomposition 522:534 arg1 detection 547:555 beam-type collision induced decomposition (beam-CID) detection 494:555 beam-type collision induced decomposition (beam-CID) detection 494:555 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 0 2 theme Integrated 67:76 arg1 Strategy 78:85 an Integrated Strategy 64:85 an Integrated Strategy 64:85 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 0 3 from Analysis 11:18 arg1 Serum 55:59 Human Serum 49:59 Human Serum 49:59 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 6 4 gly glycopeptides 1140:1152 arg2 glycopeptides 1140:1152 the glycopeptides 1136:1152 the glycopeptides 1136:1152 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 6 5 theme glycopeptides 1140:1152 arg1 % 1131:1131 About 81% 1123:1131 About 81% of the glycopeptides 1123:1152 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 6 5 theme glycopeptides 1140:1152 arg1 glycopeptides 1140:1152 the glycopeptides 1136:1152 the glycopeptides 1136:1152 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 7 6 theme proteome 1371:1378 arg1 level 1380:1384 the proteome level 1367:1384 the proteome level 1367:1384 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 4 7 theme multiple 866:873 arg1 modifications 889:901 multiple glycosylation modifications 866:901 multiple glycosylation modifications 866:901 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 4 8 theme in 790:791 arg1 method 816:821 an in silico deglycosylation method 787:821 an in silico deglycosylation method 787:821 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 3 9 theme deglycosylation 572:586 arg1 method 588:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method 431:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation 431:620 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 0 10 from Serum 55:59 arg1 Analysis 11:18 Proteomics Analysis 0:18 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 0:86 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 5 11 theme human 1024:1028 arg1 serum 1030:1034 human serum 1024:1034 human serum 1024:1034 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 7 12 theme largest 1285:1291 arg1 data 1293:1296 the largest data 1281:1296 the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level 1281:1384 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 7 12 theme largest 1285:1291 arg1 this 1273:1276 this 1273:1276 this 1273:1276 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 3 13 theme in 562:563 arg1 deglycosylation 572:586 in silico deglycosylation 562:586 in silico deglycosylation 562:586 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 5 14 from identification 1048:1061 arg1 glycoproteins 1108:1120 93 glycoproteins 1105:1120 93 glycoproteins 1105:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 3 15 dep in 562:563 arg1 silico 565:570 silico 565:570 silico 565:570 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 16 theme integrated 331:340 arg1 strategy 342:349 an integrated strategy 328:349 an integrated strategy 328:349 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 17 theme collision 504:512 arg1 detection 547:555 beam-type collision induced decomposition (beam-CID) detection 494:555 beam-type collision induced decomposition (beam-CID) detection 494:555 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 4 18 theme intact 645:650 arg1 glycopeptides 661:673 the intact O-GalNAc glycopeptides 641:673 the intact O-GalNAc glycopeptides 641:673 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 3 19 theme detection 547:555 arg1 method 588:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method 431:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation 431:620 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 20 theme hydrophilic 431:441 arg1 chromatography 455:468 hydrophilic interaction chromatography 431:468 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 20 theme hydrophilic 431:441 arg1 HILIC 471:475 HILIC 471:475 HILIC 471:475 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 21 theme chromatography 455:468 arg1 enrichment 482:491 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 0 22 theme Human 49:53 arg1 Serum 55:59 Human Serum 49:59 Human Serum 49:59 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 4 23 theme glycosylation 875:887 arg1 modifications 889:901 multiple glycosylation modifications 866:901 multiple glycosylation modifications 866:901 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 5 24 theme intact 1070:1075 arg1 glycopeptides 1086:1098 407 intact O-GalNAc glycopeptides 1066:1098 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1066:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 0 25 from Glycosylation 32:44 arg1 Serum 55:59 Human Serum 49:59 Human Serum 49:59 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 4 26 gly glycopeptides 661:673 arg2 glycopeptides 661:673 the intact O-GalNAc glycopeptides 641:673 the intact O-GalNAc glycopeptides 641:673 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 3 27 theme glycosylation 404:416 arg1 analysis 383:390 comprehensive analysis 369:390 comprehensive analysis of O-GalNAc glycosylation 369:416 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 7 28 theme intact 1305:1310 arg1 glycoforms 1321:1330 intact O-GalNAc glycoforms 1305:1330 intact O-GalNAc glycoforms 1305:1330 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 1 29 theme vital 174:178 arg1 roles 191:195 its vital biological roles 170:195 its vital biological roles 170:195 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 0 30 theme Proteomics 0:9 arg1 Analysis 11:18 Proteomics Analysis 0:18 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 0:86 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 2 31 with O-glycopeptides 214:228 arg1 glycans 245:251 different glycans 235:251 different glycans 235:251 However, intact O-glycopeptides with different glycans are typically not well elucidated using the current methods. 28035807 4 32 theme TOF 754:756 arg1 -CID 758:761 time-of-flight (TOF)-CID 738:761 time-of-flight (TOF)-CID 738:761 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 6 33 gly microheterogeneity 1213:1230 arg1 glycosylation 1244:1256 O-GalNAc glycosylation 1235:1256 O-GalNAc glycosylation 1235:1256 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 3 34 theme interaction 443:453 arg1 chromatography 455:468 hydrophilic interaction chromatography 431:468 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 34 theme interaction 443:453 arg1 HILIC 471:475 HILIC 471:475 HILIC 471:475 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 0 35 theme O-GalNAc 23:30 arg1 Glycosylation 32:44 O-GalNAc Glycosylation 23:44 O-GalNAc Glycosylation in Human Serum 23:59 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 8 36 theme biological 1486:1495 arg1 systems 1497:1503 other complex biological systems 1472:1503 other complex biological systems 1472:1503 Furthermore, this method is readily applicable to study O-glycoform heterogeneity in other complex biological systems. 28035807 5 37 theme glycopeptides 1086:1098 arg1 identification 1048:1061 identification 1048:1061 identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1048:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 5 38 theme O-GalNAc 998:1005 arg1 glycoproteome 1007:1019 the O-GalNAc glycoproteome 994:1019 the O-GalNAc glycoproteome of human serum 994:1034 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 2 39 theme different 235:243 arg1 glycans 245:251 different glycans 235:251 different glycans 235:251 However, intact O-glycopeptides with different glycans are typically not well elucidated using the current methods. 28035807 3 40 theme tip 478:480 arg1 enrichment 482:491 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 hydrophilic interaction chromatography (HILIC) tip enrichment 431:491 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 41 theme comprehensive 369:381 arg1 analysis 383:390 comprehensive analysis 369:390 comprehensive analysis of O-GalNAc glycosylation 369:416 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 4 42 theme deglycosylation 800:814 arg1 method 816:821 an in silico deglycosylation method 787:821 an in silico deglycosylation method 787:821 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 6 43 theme O-GalNAc 1235:1242 arg1 glycosylation 1244:1256 O-GalNAc glycosylation 1235:1256 O-GalNAc glycosylation 1235:1256 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 2 44 theme current 297:303 arg1 methods 305:311 the current methods 293:311 the current methods 293:311 However, intact O-glycopeptides with different glycans are typically not well elucidated using the current methods. 28035807 5 45 theme O-GalNAc 1077:1084 arg1 glycopeptides 1086:1098 407 intact O-GalNAc glycopeptides 1066:1098 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1066:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 3 46 theme enrichment 482:491 arg1 method 588:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method 431:593 hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation 431:620 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 3 47 theme induced 514:520 arg1 detection 547:555 beam-type collision induced decomposition (beam-CID) detection 494:555 beam-type collision induced decomposition (beam-CID) detection 494:555 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 6 48 theme sialic 1177:1182 arg1 acid 1184:1187 at least one sialic acid 1164:1187 at least one sialic acid 1164:1187 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 1 49 dep vital 174:178 arg1 biological 180:189 biological 180:189 biological 180:189 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 0 50 theme Glycosylation 32:44 arg1 Analysis 11:18 Proteomics Analysis 0:18 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 0:86 Proteomics Analysis of O-GalNAc Glycosylation in Human Serum by an Integrated Strategy. 28035807 5 51 gly glycopeptides 1086:1098 arg2 glycopeptides 1086:1098 407 intact O-GalNAc glycopeptides 1066:1098 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1066:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 5 51 gly glycopeptides 1086:1098 arg1 glycoproteins 1108:1120 93 glycoproteins 1105:1120 93 glycoproteins 1105:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 7 52 theme complex 1337:1343 arg1 samples 1356:1362 complex biological samples 1337:1362 complex biological samples at the proteome level 1337:1384 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 3 53 theme O-GalNAc 395:402 arg1 glycosylation 404:416 O-GalNAc glycosylation 395:416 O-GalNAc glycosylation 395:416 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 7 54 gly glycoforms 1321:1330 arg1 O-GalNAc 1312:1319 intact O-GalNAc glycoforms 1305:1330 intact O-GalNAc glycoforms 1305:1330 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 3 55 theme spectra 599:605 arg1 interpretation 607:620 spectra interpretation 599:620 spectra interpretation 599:620 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 1 56 theme O-linked 105:112 arg1 structures 121:130 O-linked glycan structures 105:130 O-linked glycan structures 105:130 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 2 57 gly O-glycopeptides 214:228 arg2 O-glycopeptides 214:228 intact O-glycopeptides 207:228 intact O-glycopeptides with different glycans 207:251 However, intact O-glycopeptides with different glycans are typically not well elucidated using the current methods. 28035807 4 58 theme search 943:948 arg1 space 950:954 the search space 939:954 the search space 939:954 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 2 59 theme intact 207:212 arg1 O-glycopeptides 214:228 intact O-glycopeptides 207:228 intact O-glycopeptides with different glycans 207:251 However, intact O-glycopeptides with different glycans are typically not well elucidated using the current methods. 28035807 7 60 theme O-GalNAc 1312:1319 arg1 glycoforms 1321:1330 intact O-GalNAc glycoforms 1305:1330 intact O-GalNAc glycoforms 1305:1330 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 1 61 theme glycan 114:119 arg1 structures 121:130 O-linked glycan structures 105:130 O-linked glycan structures 105:130 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 8 62 from heterogeneity 1455:1467 arg1 systems 1497:1503 other complex biological systems 1472:1503 other complex biological systems 1472:1503 Furthermore, this method is readily applicable to study O-glycoform heterogeneity in other complex biological systems. 28035807 8 63 theme complex 1478:1484 arg1 systems 1497:1503 other complex biological systems 1472:1503 other complex biological systems 1472:1503 Furthermore, this method is readily applicable to study O-glycoform heterogeneity in other complex biological systems. 28035807 1 64 link O-linked 105:112 arg1 structures 121:130 O-linked glycan structures 105:130 O-linked glycan structures 105:130 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 8 65 theme other 1472:1476 arg1 systems 1497:1503 other complex biological systems 1472:1503 other complex biological systems 1472:1503 Furthermore, this method is readily applicable to study O-glycoform heterogeneity in other complex biological systems. 28035807 4 66 theme original 709:716 arg1 spectra 718:724 the original spectra 705:724 the original spectra obtained by time-of-flight (TOF)-CID 705:761 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 1 67 theme structures 121:130 arg1 diversity 92:100 The diversity 88:100 The diversity of O-linked glycan structures 88:130 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 6 68 contain contained 1154:1162 arg2 acid 1184:1187 at least one sialic acid 1164:1187 at least one sialic acid 1164:1187 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 6 68 contain contained 1154:1162 arg1 % 1131:1131 About 81% 1123:1131 About 81% of the glycopeptides 1123:1152 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 6 68 contain contained 1154:1162 arg1 glycopeptides 1140:1152 the glycopeptides 1136:1152 the glycopeptides 1136:1152 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 5 69 theme serum 1030:1034 arg1 glycoproteome 1007:1019 the O-GalNAc glycoproteome 994:1019 the O-GalNAc glycoproteome of human serum 994:1034 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 3 70 theme beam-type 494:502 arg1 detection 547:555 beam-type collision induced decomposition (beam-CID) detection 494:555 beam-type collision induced decomposition (beam-CID) detection 494:555 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 5 71 from glycoproteins 1108:1120 arg1 identification 1048:1061 identification 1048:1061 identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1048:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 5 71 from glycoproteins 1108:1120 arg1 glycopeptides 1086:1098 407 intact O-GalNAc glycopeptides 1066:1098 407 intact O-GalNAc glycopeptides from 93 glycoproteins 1066:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 5 72 gly glycoproteins 1108:1120 arg1 glycoproteins 1108:1120 93 glycoproteins 1105:1120 93 glycoproteins 1105:1120 This strategy was applied to analyze the O-GalNAc glycoproteome of human serum, leading to identification of 407 intact O-GalNAc glycopeptides from 93 glycoproteins. 28035807 4 73 theme direct 833:838 arg1 searching 840:848 direct searching 833:848 direct searching 833:848 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 4 74 theme O-GalNAc 652:659 arg1 glycopeptides 661:673 the intact O-GalNAc glycopeptides 641:673 the intact O-GalNAc glycopeptides 641:673 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 3 75 theme beam-CID 537:544 arg1 detection 547:555 beam-type collision induced decomposition (beam-CID) detection 494:555 beam-type collision induced decomposition (beam-CID) detection 494:555 In this work, an integrated strategy was developed for comprehensive analysis of O-GalNAc glycosylation by combining hydrophilic interaction chromatography (HILIC) tip enrichment, beam-type collision induced decomposition (beam-CID) detection, and in silico deglycosylation method for spectra interpretation. 28035807 7 76 theme biological 1345:1354 arg1 samples 1356:1362 complex biological samples 1337:1362 complex biological samples at the proteome level 1337:1384 Up until now, this is the largest data set of intact O-GalNAc glycoforms from complex biological samples at the proteome level. 28035807 4 77 dep in 790:791 arg1 silico 793:798 silico 793:798 silico 793:798 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 28035807 6 78 theme glycosylation 1244:1256 arg1 microheterogeneity 1213:1230 the microheterogeneity 1209:1230 the microheterogeneity of O-GalNAc glycosylation 1209:1256 About 81% of the glycopeptides contained at least one sialic acid, which could reveal the microheterogeneity of O-GalNAc glycosylation. 28035807 1 79 theme increasing 142:151 arg1 attention 153:161 increasing attention 142:161 increasing attention 142:161 The diversity of O-linked glycan structures has drawn increasing attention due to its vital biological roles. 28035807 8 80 theme O-glycoform 1443:1453 arg1 heterogeneity 1455:1467 O-glycoform heterogeneity 1443:1467 O-glycoform heterogeneity in other complex biological systems 1443:1503 Furthermore, this method is readily applicable to study O-glycoform heterogeneity in other complex biological systems. 28035807 4 81 theme time-of-flight 738:751 arg1 -CID 758:761 time-of-flight (TOF)-CID 738:761 time-of-flight (TOF)-CID 738:761 In this strategy, the intact O-GalNAc glycopeptides were selectively enriched and the original spectra obtained by time-of-flight (TOF)-CID were preprocessed using an in silico deglycosylation method, enabling direct searching without setting multiple glycosylation modifications, which could significantly decrease the search space. 25601700 9 0 theme different 1653:1661 arg1 chains 1668:1673 the different side chains 1649:1673 the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer 1649:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 7 1 theme cross-ring 1270:1279 arg1 ions 1290:1293 Relevant cross-ring cleavage ions 1261:1293 Relevant cross-ring cleavage ions from (0,2)X(j)-type 1261:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 11 2 theme subtle 2125:2130 arg1 changes 2143:2149 the subtle structural changes 2121:2149 the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2121:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 3 theme quadrupole 597:606 arg1 Fourier 608:614 a hybrid quadrupole Fourier 588:614 a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 588:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 4 from column 353:358 arg1 less 363:366 less 363:366 less 363:366 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 9 5 theme MS 1685:1686 arg1 capabilities 1691:1702 the MS(n) capabilities 1681:1702 the MS(n) capabilities of the LTQ/Orbitrap analyzer 1681:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 11 6 theme ionization 2051:2060 arg1 mode 2062:2065 negative ionization mode 2042:2065 negative ionization mode 2042:2065 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 4 7 theme ionized 893:899 arg1 GalMos 901:906 the ionized GalMos 889:906 the ionized GalMos 889:906 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 7 8 theme acid 1427:1430 arg1 unit 1432:1435 the 1-2-linked glucuronic acid unit 1401:1435 the 1-2-linked glucuronic acid unit 1401:1435 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 11 9 theme cell 2264:2267 arg1 mutants 2274:2280 cell wall mutants 2264:2280 cell wall mutants with altered hemicellulose structure 2264:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 10 theme ionization 468:477 arg1 spectrometry 484:495 negative electrospray ionization mass spectrometry 446:495 negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 446:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 11 theme cyclotron 630:638 arg1 spectrometers 664:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 7 12 theme 1-2-linked 1405:1414 arg1 unit 1432:1435 the 1-2-linked glucuronic acid unit 1401:1435 the 1-2-linked glucuronic acid unit 1401:1435 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 9 13 theme analyzer 1724:1731 arg1 capabilities 1691:1702 the MS(n) capabilities 1681:1702 the MS(n) capabilities of the LTQ/Orbitrap analyzer 1681:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 2 14 theme PA 346:347 arg1 column 353:358 CarboPac PA 200 column 337:358 CarboPac PA 200 column in less than 15 min 337:378 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 11 15 theme useful 2101:2106 arg1 technique 2091:2099 a relevant and powerful technique 2067:2099 a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2067:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 6 16 theme main 1168:1171 arg1 backbone 1186:1193 the main glycosylated backbone 1164:1193 the main glycosylated backbone 1164:1193 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 9 17 theme chains 1668:1673 arg1 fragmentation 1632:1644 the fragmentation 1628:1644 the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer 1628:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 2 18 theme trap 564:567 arg1 /Orbitrap 574:582 a hybrid linear ion trap (LTQ)/Orbitrap 544:582 a hybrid linear ion trap (LTQ)/Orbitrap 544:582 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 11 19 theme organs 2202:2207 arg1 development 2181:2191 the development 2177:2191 the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2177:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 9 20 theme LTQ/Orbitrap 1711:1722 arg1 analyzer 1724:1731 the LTQ/Orbitrap analyzer 1707:1731 the LTQ/Orbitrap analyzer 1707:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 7 21 theme units 1369:1373 arg1 diagnostic 1330:1339 diagnostic 1330:1339 diagnostic 1330:1339 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 5 22 theme ion 958:960 arg1 spectra 962:968 The product ion spectra 946:968 The product ion spectra 946:968 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 7 23 theme 1-2-linked- 1348:1358 arg1 units 1369:1373 the 1-2-linked- glycosyl units 1344:1373 the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans 1344:1456 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 9 24 theme differentiating 1746:1760 arg1 arabinosyl 1771:1780 differentiating terminal arabinosyl 1746:1780 differentiating terminal arabinosyl 1746:1780 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 11 25 theme structural 2132:2141 arg1 changes 2143:2149 the subtle structural changes 2121:2149 the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2121:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 8 26 from activation 1468:1477 arg1 Orbitrap 1490:1497 the LTQ Orbitrap 1482:1497 the LTQ Orbitrap 1482:1497 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 7 27 link 1-2-linked 1405:1414 arg1 unit 1432:1435 the 1-2-linked glucuronic acid unit 1401:1435 the 1-2-linked glucuronic acid unit 1401:1435 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 3 28 located observed 728:735 arg2 routes 721:726 the fragmentation routes 703:726 the fragmentation routes observed on XylGos 703:745 In order to corroborate the fragmentation routes observed on XylGos, some commercial galacto-manno-oligosaccharides (GalMOs) and glucurono-xylo-oligosaccharides were also studied. 25601700 3 28 located observed 728:735 arg1 XylGos 740:745 XylGos 740:745 XylGos 740:745 In order to corroborate the fragmentation routes observed on XylGos, some commercial galacto-manno-oligosaccharides (GalMOs) and glucurono-xylo-oligosaccharides were also studied. 25601700 2 29 theme linear 553:558 arg1 LTQ 570:572 LTQ 570:572 LTQ 570:572 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 29 theme linear 553:558 arg1 trap 564:567 a hybrid linear ion trap 544:567 a hybrid linear ion trap (LTQ)/Orbitrap 544:582 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 30 theme mass 659:662 arg1 spectrometers 664:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 5 31 theme prominent 999:1007 arg1 ions 1029:1032 prominent double cleavage (D) ions 999:1032 prominent double cleavage (D) ions corresponding to the entire inner side chains 999:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 0 32 theme linkage 92:98 arg1 position 100:107 determining linkage position 80:107 determining linkage position 80:107 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 5 33 theme cleavage 1016:1023 arg1 ions 1029:1032 prominent double cleavage (D) ions 999:1032 prominent double cleavage (D) ions corresponding to the entire inner side chains 999:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 11 34 theme wall 2269:2272 arg1 mutants 2274:2280 cell wall mutants 2264:2280 cell wall mutants with altered hemicellulose structure 2264:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 35 theme FT-ICR 651:656 arg1 spectrometers 664:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 9 36 theme XylGos 1837:1842 arg1 side-chains 1822:1832 U side-chains 1820:1832 U side-chains of XylGos 1820:1842 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 9 36 theme XylGos 1837:1842 arg1 S 1814:1814 S 1814:1814 S 1814:1814 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 5 37 theme D 1026:1026 arg1 ions 1029:1032 prominent double cleavage (D) ions 999:1032 prominent double cleavage (D) ions corresponding to the entire inner side chains 999:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 11 38 theme powerful 2082:2089 arg1 technique 2091:2099 a relevant and powerful technique 2067:2099 a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2067:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 39 theme mass 479:482 arg1 spectrometry 484:495 negative electrospray ionization mass spectrometry 446:495 negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 446:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 0 40 theme Negative 0:7 arg1 spectrometry 38:49 Negative electrospray ionization mass spectrometry 0:49 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 0:157 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 8 41 dep allowed 1499:1505 arg1 only 1511:1514 only 1511:1514 only 1511:1514 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 2 42 theme electrospray 455:466 arg1 spectrometry 484:495 negative electrospray ionization mass spectrometry 446:495 negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 446:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 0 43 theme ionization 22:31 arg1 spectrometry 38:49 Negative electrospray ionization mass spectrometry 0:49 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 0:157 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 8 44 gly fucosylated 1593:1603 arg1 side-chains 1605:1615 fucosylated side-chains 1593:1615 fucosylated side-chains 1593:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 7 45 from XylGos 1380:1385 arg1 units 1369:1373 the 1-2-linked- glycosyl units 1344:1373 the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans 1344:1456 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 2 46 theme quadrupole 505:514 arg1 ESI-Q-TOF 532:540 ESI-Q-TOF 532:540 ESI-Q-TOF 532:540 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 46 theme quadrupole 505:514 arg1 time-of-flight 516:529 a quadrupole time-of-flight 503:529 a quadrupole time-of-flight (ESI-Q-TOF) 503:541 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 4 47 theme fragmentation 863:875 arg1 pathways 877:884 The fragmentation pathways 859:884 The fragmentation pathways of the ionized GalMos 859:906 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 4 47 theme fragmentation 863:875 arg1 similar 913:919 similar 913:919 similar 913:919 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 5 48 theme entire 1055:1060 arg1 chains 1073:1078 the entire inner side chains 1051:1078 the entire inner side chains 1051:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 7 49 theme j 1307:1307 arg1 -type 1309:1313 (0,2)X(j)-type 1300:1313 (0,2)X(j)-type 1300:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 11 50 theme fruit 2224:2228 arg1 ripening 2230:2237 fruit ripening 2224:2237 fruit ripening 2224:2237 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 8 51 theme group 1575:1579 arg1 location 1581:1588 the O-acetyl group location 1562:1588 the type of all linkages but also the O-acetyl group location on fucosylated side-chains 1528:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 7 52 theme Relevant 1261:1268 arg1 ions 1290:1293 Relevant cross-ring cleavage ions 1261:1293 Relevant cross-ring cleavage ions from (0,2)X(j)-type 1261:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 8 53 from type 1532:1535 arg1 side-chains 1605:1615 fucosylated side-chains 1593:1615 fucosylated side-chains 1593:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 5 54 theme side 1068:1071 arg1 chains 1073:1078 the entire inner side chains 1051:1078 the entire inner side chains 1051:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 7 55 theme X 1305:1305 arg1 -type 1309:1313 (0,2)X(j)-type 1300:1313 (0,2)X(j)-type 1300:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 1 56 theme commercial 230:239 arg1 β-1-4-D-endoglucanase 246:266 commercial endo β-1-4-D-endoglucanase 230:266 commercial endo β-1-4-D-endoglucanase 230:266 Xyloglucans of apple, tomato, bilberry and tamarind were hydrolyzed by commercial endo β-1-4-D-endoglucanase. 25601700 7 57 theme 0,2 1301:1303 arg1 -type 1309:1313 (0,2)X(j)-type 1300:1313 (0,2)X(j)-type 1300:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 0 58 from hemicelluloses 143:156 arg1 position 100:107 determining linkage position 80:107 determining linkage position 80:107 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 0 58 from hemicelluloses 143:156 arg1 sequencing 65:74 sequencing 65:74 sequencing 65:74 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 0 58 from hemicelluloses 143:156 arg1 oligosaccharides 112:127 oligosaccharides 112:127 oligosaccharides from branched hemicelluloses 112:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 6 59 from end 1126:1128 arg1 fragmentation 1094:1106 The directed fragmentation 1081:1106 The directed fragmentation from the reducing end to the other end 1081:1145 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 11 60 from fragmentation 2025:2037 arg1 mode 2062:2065 negative ionization mode 2042:2065 negative ionization mode 2042:2065 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 10 61 theme isomeric 1925:1932 arg1 structures 1934:1943 isomeric structures 1925:1943 isomeric structures differing only in their substitution pattern 1925:1988 The CID spectra obtained were very informative for distinction of isomeric structures differing only in their substitution pattern. 25601700 9 62 theme side 1663:1666 arg1 chains 1668:1673 the different side chains 1649:1673 the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer 1649:1731 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 7 63 theme cleavage 1281:1288 arg1 ions 1290:1293 Relevant cross-ring cleavage ions 1261:1293 Relevant cross-ring cleavage ions from (0,2)X(j)-type 1261:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 11 64 theme negative 2042:2049 arg1 mode 2062:2065 negative ionization mode 2042:2065 negative ionization mode 2042:2065 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 65 theme hybrid 590:595 arg1 Fourier 608:614 a hybrid quadrupole Fourier 588:614 a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 588:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 66 theme resonance 640:648 arg1 spectrometers 664:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 11 67 theme hemicellulose 2295:2307 arg1 structure 2309:2317 altered hemicellulose structure 2287:2317 altered hemicellulose structure 2287:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 6 68 gly glycosylated 1173:1184 arg1 backbone 1186:1193 the main glycosylated backbone 1164:1193 the main glycosylated backbone 1164:1193 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 8 69 from location 1581:1588 arg1 side-chains 1605:1615 fucosylated side-chains 1593:1615 fucosylated side-chains 1593:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 11 70 theme relevant 2069:2076 arg1 technique 2091:2099 a relevant and powerful technique 2067:2099 a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2067:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 2 71 theme ion 626:628 arg1 spectrometers 664:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 ion cyclotron resonance (FT-ICR) mass spectrometers 626:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 7 72 theme glucuronic 1416:1425 arg1 unit 1432:1435 the 1-2-linked glucuronic acid unit 1401:1435 the 1-2-linked glucuronic acid unit 1401:1435 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 0 73 theme branched 134:141 arg1 hemicelluloses 143:156 branched hemicelluloses 134:156 branched hemicelluloses 134:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 0 74 dep spectrometry 38:49 arg1 method 54:59 a method 52:59 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 0:157 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 2 75 theme CarboPac 337:344 arg1 column 353:358 CarboPac PA 200 column 337:358 CarboPac PA 200 column in less than 15 min 337:378 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 6 76 theme glycosylated 1173:1184 arg1 backbone 1186:1193 the main glycosylated backbone 1164:1193 the main glycosylated backbone 1164:1193 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 8 77 theme Resonant 1459:1466 arg1 activation 1468:1477 Resonant activation 1459:1477 Resonant activation in the LTQ Orbitrap 1459:1497 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 3 78 theme commercial 753:762 arg1 galacto-manno-oligosaccharides 764:793 some commercial galacto-manno-oligosaccharides 748:793 some commercial galacto-manno-oligosaccharides (GalMOs) 748:802 In order to corroborate the fragmentation routes observed on XylGos, some commercial galacto-manno-oligosaccharides (GalMOs) and glucurono-xylo-oligosaccharides were also studied. 25601700 3 78 theme commercial 753:762 arg1 GalMOs 796:801 GalMOs 796:801 GalMOs 796:801 In order to corroborate the fragmentation routes observed on XylGos, some commercial galacto-manno-oligosaccharides (GalMOs) and glucurono-xylo-oligosaccharides were also studied. 25601700 2 79 theme ion 560:562 arg1 LTQ 570:572 LTQ 570:572 LTQ 570:572 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 79 theme ion 560:562 arg1 trap 564:567 a hybrid linear ion trap 544:567 a hybrid linear ion trap (LTQ)/Orbitrap 544:582 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 0 80 from sequencing 65:74 arg1 hemicelluloses 143:156 branched hemicelluloses 134:156 branched hemicelluloses 134:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 0 80 from sequencing 65:74 arg1 oligosaccharides 112:127 oligosaccharides 112:127 oligosaccharides from branched hemicelluloses 112:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 11 81 with mutants 2274:2280 arg1 structure 2309:2317 altered hemicellulose structure 2287:2317 altered hemicellulose structure 2287:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 9 82 theme xylosyl 1786:1792 arg1 substituents 1794:1805 xylosyl substituents 1786:1805 xylosyl substituents 1786:1805 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 5 83 theme product 950:956 arg1 spectra 962:968 The product ion spectra 946:968 The product ion spectra 946:968 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 2 84 dep Fourier 608:614 arg1 transform 616:624 transform 616:624 transform ion cyclotron resonance (FT-ICR) mass spectrometers 616:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 4 85 theme XylGos 933:938 arg1 ones 940:943 XylGos ones 933:943 XylGos ones 933:943 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 6 86 theme complete 1240:1247 arg1 sequencing 1249:1258 their complete sequencing 1234:1258 their complete sequencing 1234:1258 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 0 87 from position 100:107 arg1 hemicelluloses 143:156 branched hemicelluloses 134:156 branched hemicelluloses 134:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 0 87 from position 100:107 arg1 oligosaccharides 112:127 oligosaccharides 112:127 oligosaccharides from branched hemicelluloses 112:156 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 7 88 theme glycosyl 1360:1367 arg1 units 1369:1373 the 1-2-linked- glycosyl units 1344:1373 the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans 1344:1456 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 9 89 theme terminal 1762:1769 arg1 arabinosyl 1771:1780 differentiating terminal arabinosyl 1746:1780 differentiating terminal arabinosyl 1746:1780 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 11 90 theme altered 2287:2293 arg1 structure 2309:2317 altered hemicellulose structure 2287:2317 altered hemicellulose structure 2287:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 0 91 theme determining 80:90 arg1 position 100:107 determining linkage position 80:107 determining linkage position 80:107 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 5 92 theme double 1009:1014 arg1 ions 1029:1032 prominent double cleavage (D) ions 999:1032 prominent double cleavage (D) ions corresponding to the entire inner side chains 999:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 2 93 theme hybrid 546:551 arg1 LTQ 570:572 LTQ 570:572 LTQ 570:572 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 2 93 theme hybrid 546:551 arg1 trap 564:567 a hybrid linear ion trap 544:567 a hybrid linear ion trap (LTQ)/Orbitrap 544:582 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 6 94 theme reducing 1117:1124 arg1 end 1126:1128 the reducing end 1113:1128 the reducing end 1113:1128 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 11 95 theme mutants 2274:2280 arg1 screening 2251:2259 the screening 2247:2259 the screening of cell wall mutants with altered hemicellulose structure 2247:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 8 96 theme linkages 1544:1551 arg1 location 1581:1588 the O-acetyl group location 1562:1588 the type of all linkages but also the O-acetyl group location on fucosylated side-chains 1528:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 8 96 theme linkages 1544:1551 arg1 type 1532:1535 the type 1528:1535 the type of all linkages but also the O-acetyl group location on fucosylated side-chains 1528:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 11 97 theme plant 2196:2200 arg1 ripening 2230:2237 fruit ripening 2224:2237 fruit ripening 2224:2237 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 11 97 theme plant 2196:2200 arg1 organs 2202:2207 plant organs 2196:2207 plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure 2196:2317 These features together makes the fragmentation in negative ionization mode a relevant and powerful technique useful to highlight the subtle structural changes generally observed during the development of plant organs such as during fruit ripening and for the screening of cell wall mutants with altered hemicellulose structure. 25601700 0 98 theme electrospray 9:20 arg1 spectrometry 38:49 Negative electrospray ionization mass spectrometry 0:49 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 0:157 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 10 99 theme substitution 1969:1980 arg1 pattern 1982:1988 their substitution pattern 1963:1988 their substitution pattern 1963:1988 The CID spectra obtained were very informative for distinction of isomeric structures differing only in their substitution pattern. 25601700 9 100 theme U 1820:1820 arg1 side-chains 1822:1832 U side-chains 1820:1832 U side-chains of XylGos 1820:1842 Moreover, the fragmentation of the different side chains using the MS(n) capabilities of the LTQ/Orbitrap analyzer also allowed differentiating terminal arabinosyl and xylosyl substituents inside S and U side-chains of XylGos, respectively. 25601700 0 101 theme mass 33:36 arg1 spectrometry 38:49 Negative electrospray ionization mass spectrometry 0:49 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 0:157 Negative electrospray ionization mass spectrometry: a method for sequencing and determining linkage position in oligosaccharides from branched hemicelluloses. 25601700 6 102 theme directed 1085:1092 arg1 fragmentation 1094:1106 The directed fragmentation 1081:1106 The directed fragmentation from the reducing end to the other end 1081:1145 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 2 103 theme negative 446:453 arg1 spectrometry 484:495 negative electrospray ionization mass spectrometry 446:495 negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers 446:676 The xylo-gluco-oligosaccharides (XylGos) released were separated on CarboPac PA 200 column in less than 15 min, and, after purification, they were structurally characterized by negative electrospray ionization mass spectrometry using a quadrupole time-of-flight (ESI-Q-TOF), a hybrid linear ion trap (LTQ)/Orbitrap and a hybrid quadrupole Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometers. 25601700 8 104 theme O-acetyl 1566:1573 arg1 location 1581:1588 the O-acetyl group location 1562:1588 the type of all linkages but also the O-acetyl group location on fucosylated side-chains 1528:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 10 105 theme CID 1863:1865 arg1 spectra 1867:1873 The CID spectra 1859:1873 The CID spectra obtained 1859:1882 The CID spectra obtained were very informative for distinction of isomeric structures differing only in their substitution pattern. 25601700 10 105 theme CID 1863:1865 arg1 informative 1894:1904 informative 1894:1904 informative 1894:1904 The CID spectra obtained were very informative for distinction of isomeric structures differing only in their substitution pattern. 25601700 5 106 theme inner 1062:1066 arg1 chains 1073:1078 the entire inner side chains 1051:1078 the entire inner side chains 1051:1078 The product ion spectra were mainly characterized by prominent double cleavage (D) ions corresponding to the entire inner side chains. 25601700 4 107 theme GalMos 901:906 arg1 pathways 877:884 The fragmentation pathways 859:884 The fragmentation pathways of the ionized GalMos 859:906 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 4 107 theme GalMos 901:906 arg1 similar 913:919 similar 913:919 similar 913:919 The fragmentation pathways of the ionized GalMos were similar to those of XylGos ones. 25601700 8 108 theme fucosylated 1593:1603 arg1 side-chains 1605:1615 fucosylated side-chains 1593:1615 fucosylated side-chains 1593:1615 Resonant activation in the LTQ Orbitrap allowed not only determining the type of all linkages but also the O-acetyl group location on fucosylated side-chains. 25601700 10 109 theme structures 1934:1943 arg1 distinction 1910:1920 distinction 1910:1920 distinction of isomeric structures differing only in their substitution pattern 1910:1988 The CID spectra obtained were very informative for distinction of isomeric structures differing only in their substitution pattern. 25601700 7 110 from -type 1309:1313 arg1 ions 1290:1293 Relevant cross-ring cleavage ions 1261:1293 Relevant cross-ring cleavage ions from (0,2)X(j)-type 1261:1313 Relevant cross-ring cleavage ions from (0,2)X(j)-type revealed to be diagnostic of the 1-2-linked- glycosyl units from XylGos together with the 1-2-linked glucuronic acid unit from glucuronoxylans. 25601700 1 111 theme endo 241:244 arg1 β-1-4-D-endoglucanase 246:266 commercial endo β-1-4-D-endoglucanase 230:266 commercial endo β-1-4-D-endoglucanase 230:266 Xyloglucans of apple, tomato, bilberry and tamarind were hydrolyzed by commercial endo β-1-4-D-endoglucanase. 25601700 6 112 theme other 1137:1141 arg1 end 1143:1145 the other end 1133:1145 the other end 1133:1145 The directed fragmentation from the reducing end to the other end was observed for the main glycosylated backbone but also for the side-chains, allowing their complete sequencing. 25601700 3 113 theme fragmentation 707:719 arg1 routes 721:726 the fragmentation routes 703:726 the fragmentation routes observed on XylGos 703:745 In order to corroborate the fragmentation routes observed on XylGos, some commercial galacto-manno-oligosaccharides (GalMOs) and glucurono-xylo-oligosaccharides were also studied. 25416425 5 0 theme protein 803:809 arg1 expression 811:820 surface protein expression 795:820 surface protein expression of Kv1.4 795:829 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 6 1 theme total 949:953 arg1 level 963:967 total protein level 949:967 total protein level 949:967 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 9 2 theme Kv1.4 1214:1218 arg1 level 1236:1240 Kv1.4 surface protein level 1214:1240 Kv1.4 surface protein level 1214:1240 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 1 3 theme glycosylated 155:166 arg1 channels 134:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 1 3 theme glycosylated 155:166 arg1 proteins 168:175 heavily glycosylated proteins 147:175 heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity 147:257 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 0 4 theme expression 71:80 arg1 levels 82:87 the trafficking and cell surface expression levels 38:87 the trafficking and cell surface expression levels of Kv1.4 potassium channels 38:115 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 10 5 theme surface 1497:1503 arg1 expression 1505:1514 high cell surface expression 1487:1514 high cell surface expression of Kv1.4 1487:1523 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 9 6 theme surface 1220:1226 arg1 level 1236:1240 Kv1.4 surface protein level 1214:1240 Kv1.4 surface protein level 1214:1240 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 3 7 theme cell 498:501 arg1 expression 511:520 Kv1.4 cell surface expression 492:520 Kv1.4 cell surface expression 492:520 Here we investigated whether the composition of the N-glycan affected Kv1.4 cell surface expression. 25416425 0 8 theme Kv1.4 92:96 arg1 channels 108:115 Kv1.4 potassium channels 92:115 Kv1.4 potassium channels 92:115 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 10 9 theme cell 1492:1495 arg1 expression 1505:1514 high cell surface expression 1487:1514 high cell surface expression of Kv1.4 1487:1523 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 9 10 theme protein 1228:1234 arg1 level 1236:1240 Kv1.4 surface protein level 1214:1240 Kv1.4 surface protein level 1214:1240 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 2 11 dep exhibited 294:302 arg1 Watanabe 369:376 Watanabe 369:376 Watanabe 369:376 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 8 12 from effect 1157:1162 arg1 expression 1180:1189 cell surface expression 1167:1189 cell surface expression of Kv1.4 1167:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 7 13 contain carried 1039:1045 arg2 N-glycan 1074:1081 a unique oligomannose-type N-glycan 1047:1081 a unique oligomannose-type N-glycan that contains three glucose residues 1047:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 7 13 contain carried 1039:1045 arg1 Kv1.4 1011:1015 Kv1.4 1011:1015 Kv1.4 in CSTP-treated cells 1011:1037 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 4 14 contain carrying 538:545 arg1 proteins 529:536 Kv1.4 proteins 523:536 Kv1.4 proteins carrying N-glycans with different compositions 523:583 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 4 14 contain carrying 538:545 arg2 N-glycans 547:555 N-glycans 547:555 N-glycans with different compositions 547:583 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 3 15 theme Kv1.4 492:496 arg1 expression 511:520 Kv1.4 cell surface expression 492:520 Kv1.4 cell surface expression 492:520 Here we investigated whether the composition of the N-glycan affected Kv1.4 cell surface expression. 25416425 8 16 theme surface 1172:1178 arg1 expression 1180:1189 cell surface expression 1167:1189 cell surface expression of Kv1.4 1167:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 0 17 theme channels 108:115 arg1 levels 82:87 the trafficking and cell surface expression levels 38:87 the trafficking and cell surface expression levels of Kv1.4 potassium channels 38:115 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 3 18 theme N-glycan 474:481 arg1 composition 455:465 the composition 451:465 the composition of the N-glycan 451:481 Here we investigated whether the composition of the N-glycan affected Kv1.4 cell surface expression. 25416425 7 19 from Kv1.4 1011:1015 arg1 cells 1033:1037 CSTP-treated cells 1020:1037 CSTP-treated cells 1020:1037 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 0 20 theme potassium 98:106 arg1 channels 108:115 Kv1.4 potassium channels 92:115 Kv1.4 potassium channels 92:115 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 9 21 theme total 1278:1282 arg1 level 1292:1296 total protein level 1278:1296 total protein level 1278:1296 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 2 22 dep Watanabe 369:376 arg1 al. 381:383 Watanabe et al. 369:383 Watanabe et al. in J Biol Chem 279:8879-8885, 2004 369:418 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 5 23 theme negative 776:783 arg1 effect 785:790 a negative effect 774:790 a negative effect 774:790 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 5 24 theme complex-type 845:856 arg1 N-glycans 858:866 complex-type N-glycans 845:866 complex-type N-glycans 845:866 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 5 25 contain had 770:772 arg2 effect 785:790 a negative effect 774:790 a negative effect 774:790 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 5 25 contain had 770:772 arg1 N-glycans 760:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 10 26 theme Kv1.4 1519:1523 arg1 expression 1505:1514 high cell surface expression 1487:1514 high cell surface expression of Kv1.4 1487:1523 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 9 27 theme combined 1247:1254 arg1 mechanism 1256:1264 a combined mechanism 1245:1264 a combined mechanism of reducing total protein level and increasing ER-retention 1245:1324 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 7 28 contain contains 1088:1095 arg1 N-glycan 1074:1081 a unique oligomannose-type N-glycan 1047:1081 a unique oligomannose-type N-glycan that contains three glucose residues 1047:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 7 28 contain contains 1088:1095 arg2 residues 1111:1118 three glucose residues 1097:1118 three glucose residues 1097:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 1 29 theme shaping 189:195 arg1 potentials 204:213 shaping action potentials 189:213 shaping action potentials 189:213 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 7 30 theme glucose 1103:1109 arg1 residues 1111:1118 three glucose residues 1097:1118 three glucose residues 1097:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 10 31 theme complex-type 1456:1467 arg1 N-glycan 1469:1476 a sialylated complex-type N-glycan 1443:1476 a sialylated complex-type N-glycan 1443:1476 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 8 32 theme Kv1.4 1194:1198 arg1 expression 1180:1189 cell surface expression 1167:1189 cell surface expression of Kv1.4 1167:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 1 33 theme action 197:202 arg1 potentials 204:213 shaping action potentials 189:213 shaping action potentials 189:213 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 0 34 theme N-glycosylation 14:28 arg1 degree 4:9 The degree 0:9 The degree of N-glycosylation 0:28 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 10 35 theme sialylated 1445:1454 arg1 N-glycan 1469:1476 a sialylated complex-type N-glycan 1443:1476 a sialylated complex-type N-glycan 1443:1476 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 2 36 theme protein 314:320 arg1 stability 322:330 decreased protein stability 304:330 decreased protein stability 304:330 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 4 37 theme mutant 668:673 arg1 lines 680:684 N-glycosylation-deficient mutant cell lines 642:684 N-glycosylation-deficient mutant cell lines 642:684 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 6 38 theme N-glycan 989:996 arg1 composition 998:1008 altered N-glycan composition 981:1008 altered N-glycan composition 981:1008 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 5 39 theme incomplete 736:745 arg1 N-glycans 760:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 2 40 theme decreased 304:312 arg1 stability 322:330 decreased protein stability 304:330 decreased protein stability 304:330 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 4 41 theme N-glycosylation-deficient 642:666 arg1 lines 680:684 N-glycosylation-deficient mutant cell lines 642:684 N-glycosylation-deficient mutant cell lines 642:684 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 4 42 theme different 562:570 arg1 compositions 572:583 different compositions 562:583 different compositions 562:583 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 5 43 theme complex-type 747:758 arg1 N-glycans 760:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 6 44 theme Kv1.4 910:914 arg1 level 901:905 surface protein level 885:905 surface protein level of Kv1.4 885:914 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 6 45 theme altered 981:987 arg1 composition 998:1008 altered N-glycan composition 981:1008 altered N-glycan composition 981:1008 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 10 46 theme important 1386:1394 arg1 role 1396:1399 an important role 1383:1399 an important role 1383:1399 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 5 47 theme Kv1.4 825:829 arg1 expression 811:820 surface protein expression 795:820 surface protein expression of Kv1.4 795:829 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 4 48 theme Kv1.4 523:527 arg1 proteins 529:536 Kv1.4 proteins 523:536 Kv1.4 proteins carrying N-glycans with different compositions 523:583 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 6 49 from decrease 873:880 arg1 level 901:905 surface protein level 885:905 surface protein level of Kv1.4 885:914 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 9 50 theme protein 1284:1290 arg1 level 1292:1296 total protein level 1278:1296 total protein level 1278:1296 It decreased Kv1.4 surface protein level by a combined mechanism of reducing total protein level and increasing ER-retention. 25416425 10 51 gly sialylated 1445:1454 arg1 N-glycan 1469:1476 a sialylated complex-type N-glycan 1443:1476 a sialylated complex-type N-glycan 1443:1476 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 7 52 theme unique 1049:1054 arg1 N-glycan 1074:1081 a unique oligomannose-type N-glycan 1047:1081 a unique oligomannose-type N-glycan that contains three glucose residues 1047:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 1 53 theme neuronal 222:229 arg1 excitability 231:242 neuronal excitability 222:242 neuronal excitability 222:242 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 10 54 theme high 1487:1490 arg1 expression 1505:1514 high cell surface expression 1487:1514 high cell surface expression of Kv1.4 1487:1523 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 5 55 theme oligomannose-type 701:717 arg1 N-glycans 760:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 5 56 theme surface 795:801 arg1 expression 811:820 surface protein expression 795:820 surface protein expression of Kv1.4 795:829 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 4 57 with N-glycans 547:555 arg1 compositions 572:583 different compositions 562:583 different compositions 562:583 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 2 58 theme cell 355:358 arg1 surface 360:366 the cell surface 351:366 the cell surface 351:366 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 0 59 theme trafficking 42:52 arg1 levels 82:87 the trafficking and cell surface expression levels 38:87 the trafficking and cell surface expression levels of Kv1.4 potassium channels 38:115 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 8 60 theme negative 1148:1155 arg1 effect 1157:1162 the most negative effect 1139:1162 the most negative effect on cell surface expression of Kv1.4 1139:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 6 61 theme protein 893:899 arg1 level 901:905 surface protein level 885:905 surface protein level of Kv1.4 885:914 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 8 62 contain had 1135:1137 arg1 N-glycan 1126:1133 This N-glycan 1121:1133 This N-glycan 1121:1133 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 8 62 contain had 1135:1137 arg2 effect 1157:1162 the most negative effect 1139:1162 the most negative effect on cell surface expression of Kv1.4 1139:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 1 63 theme Kv1.4 118:122 arg1 channels 134:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 1 63 theme Kv1.4 118:122 arg1 proteins 168:175 heavily glycosylated proteins 147:175 heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity 147:257 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 6 64 from reduction 936:944 arg1 level 963:967 total protein level 949:967 total protein level 949:967 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 5 65 theme hybrid-type 720:730 arg1 N-glycans 760:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 oligomannose-type, hybrid-type, or incomplete complex-type N-glycans 701:768 We found that oligomannose-type, hybrid-type, or incomplete complex-type N-glycans had a negative effect on surface protein expression of Kv1.4 compared with complex-type N-glycans. 25416425 6 66 theme surface 885:891 arg1 level 901:905 surface protein level 885:905 surface protein level of Kv1.4 885:914 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 8 67 theme cell 1167:1170 arg1 expression 1180:1189 cell surface expression 1167:1189 cell surface expression of Kv1.4 1167:1198 This N-glycan had the most negative effect on cell surface expression of Kv1.4. 25416425 1 68 theme potassium 124:132 arg1 channels 134:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 1 68 theme potassium 124:132 arg1 proteins 168:175 heavily glycosylated proteins 147:175 heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity 147:257 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 1 69 gly glycosylated 155:166 arg1 channels 134:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels 118:141 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 1 69 gly glycosylated 155:166 arg1 proteins 168:175 heavily glycosylated proteins 147:175 heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity 147:257 Kv1.4 potassium channels are heavily glycosylated proteins involved in shaping action potentials and in neuronal excitability and plasticity. 25416425 7 70 theme oligomannose-type 1056:1072 arg1 N-glycan 1074:1081 a unique oligomannose-type N-glycan 1047:1081 a unique oligomannose-type N-glycan that contains three glucose residues 1047:1118 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 7 71 theme CSTP-treated 1020:1031 arg1 cells 1033:1037 CSTP-treated cells 1020:1037 CSTP-treated cells 1020:1037 Kv1.4 in CSTP-treated cells carried a unique oligomannose-type N-glycan that contains three glucose residues. 25416425 4 72 theme glycosidase 610:620 arg1 inhibitors 622:631 glycosidase inhibitors 610:631 glycosidase inhibitors 610:631 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 0 73 theme surface 63:69 arg1 expression 71:80 cell surface expression 58:80 cell surface expression 58:80 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 4 74 theme cell 675:678 arg1 lines 680:684 N-glycosylation-deficient mutant cell lines 642:684 N-glycosylation-deficient mutant cell lines 642:684 Kv1.4 proteins carrying N-glycans with different compositions were generated by adding glycosidase inhibitors or using N-glycosylation-deficient mutant cell lines. 25416425 10 75 theme N-glycan 1368:1375 arg1 composition 1349:1359 composition 1349:1359 composition of the N-glycan 1349:1375 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25416425 3 76 theme surface 503:509 arg1 expression 511:520 Kv1.4 cell surface expression 492:520 Kv1.4 cell surface expression 492:520 Here we investigated whether the composition of the N-glycan affected Kv1.4 cell surface expression. 25416425 0 77 theme cell 58:61 arg1 expression 71:80 cell surface expression 58:80 cell surface expression 58:80 The degree of N-glycosylation affects the trafficking and cell surface expression levels of Kv1.4 potassium channels. 25416425 6 78 theme protein 955:961 arg1 level 963:967 total protein level 949:967 total protein level 949:967 The decrease in surface protein level of Kv1.4 was mainly due to a reduction in total protein level, induced by altered N-glycan composition. 25416425 2 79 theme Kv1.4 260:264 arg1 N354Q 266:270 Kv1.4 N354Q 260:270 Kv1.4 N354Q 260:270 Kv1.4 N354Q, without an N-glycan, exhibited decreased protein stability and trafficking to the cell surface (Watanabe et al. in J Biol Chem 279:8879-8885, 2004). 25416425 10 80 theme protein 1404:1410 arg1 stability 1412:1420 protein stability 1404:1420 protein stability 1404:1420 Our data suggest that composition of the N-glycan plays an important role in protein stability and trafficking, and a sialylated complex-type N-glycan promoted high cell surface expression of Kv1.4. 25776888 6 0 theme large 1156:1160 arg1 intestine>caecum 1178:1193 large intestine>small intestine>caecum 1156:1193 large intestine>small intestine>caecum 1156:1193 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 10 1 theme large 1800:1804 arg1 samples 1816:1822 large intestine samples 1800:1822 large intestine samples but not small intestine or caecum 1800:1856 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 4 2 theme pathogenic 627:636 arg1 mechanisms 648:657 the pathogenic molecular mechanisms 623:657 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 2 theme pathogenic 627:636 arg1 unknown 689:695 unknown 689:695 unknown 689:695 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 7 3 theme large 1313:1317 arg1 intestine 1319:1327 chicken large intestine 1305:1327 chicken large intestine 1305:1327 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 1 4 from children 325:332 arg1 countries 348:356 developing countries 337:356 developing countries 337:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 11 5 theme chicken 1899:1905 arg1 mucins 1918:1923 chicken intestinal mucins 1899:1923 chicken intestinal mucins 1899:1923 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 5 6 theme colorectal 923:932 arg1 line 954:957 the human colorectal adenocarcinoma cell line HCT-8 913:963 the human colorectal adenocarcinoma cell line HCT-8 913:963 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 11 7 theme C. 1985:1986 arg1 jejuni 1988:1993 C. jejuni 1985:1993 C. jejuni 1985:1993 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 4 8 from humans 678:683 arg1 mechanisms 648:657 the pathogenic molecular mechanisms 623:657 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 8 from humans 678:683 arg1 unknown 689:695 unknown 689:695 unknown 689:695 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 8 from humans 678:683 arg1 interactions 550:561 The interactions 546:561 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 1 9 theme industrialized 233:246 arg1 countries 248:256 industrialized countries 233:256 industrialized countries 233:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 10 10 theme acid 1723:1726 arg1 structures 1739:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures 1683:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes 1683:1779 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 5 11 theme jejuni 903:908 arg1 invasion 888:895 invasion 888:895 invasion 888:895 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 5 11 theme jejuni 903:908 arg1 adherence 874:882 adherence 874:882 adherence 874:882 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 7 12 attach released 1270:1277 arg1 mucins 1284:1289 mucins 1284:1289 mucins purified from chicken large intestine, small intestine, and caecum 1284:1356 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 7 12 attach released 1270:1277 arg2 O-glycans 1260:1268 O-glycans 1260:1268 O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum 1260:1356 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 11 13 theme mucins 1918:1923 arg1 characterization 1879:1894 O-glycan structural characterization 1859:1894 O-glycan structural characterization of chicken intestinal mucins 1859:1923 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 1 14 theme bacteria 145:152 arg1 jejuni 168:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 14 theme bacteria 145:152 arg1 bacteria 190:197 the primary bacteria 178:197 the primary bacteria responsible for food poisoning in industrialized countries 178:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 9 15 theme high 1582:1585 arg1 affinity 1595:1602 a high binding affinity 1580:1602 a high binding affinity to C. jejuni 1580:1615 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 6 16 theme chicken 1069:1075 arg1 intestine 1077:1085 the chicken intestine 1065:1085 the chicken intestine 1065:1085 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 7 17 theme chicken 1305:1311 arg1 intestine 1319:1327 chicken large intestine 1305:1327 chicken large intestine 1305:1327 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 7 18 theme detailed 1228:1235 arg1 analysis 1248:1255 a detailed structural analysis 1226:1255 a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum 1226:1356 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 5 19 theme cell 949:952 arg1 line 954:957 the human colorectal adenocarcinoma cell line HCT-8 913:963 the human colorectal adenocarcinoma cell line HCT-8 913:963 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 4 20 theme potential 742:750 arg1 targets 752:758 potential targets 742:758 potential targets 742:758 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 11 21 theme O-glycan 1859:1866 arg1 characterization 1879:1894 O-glycan structural characterization 1859:1894 O-glycan structural characterization of chicken intestinal mucins 1859:1923 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 5 22 theme chicken 826:832 arg1 mucin 845:849 purified chicken intestinal mucin 817:849 purified chicken intestinal mucin 817:849 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 3 23 from crypts 441:446 arg1 abundant 418:425 abundant 418:425 abundant 418:425 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 9 24 theme C. 1607:1608 arg1 jejuni 1610:1615 C. jejuni 1607:1615 C. jejuni 1607:1615 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 6 25 theme different 1044:1052 arg1 regions 1054:1060 different regions 1044:1060 different regions of the chicken intestine 1044:1085 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 0 26 from Identification 0:13 arg1 Mucins 62:67 Chicken Intestinal Mucins 43:67 Chicken Intestinal Mucins 43:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 4 27 theme campylobacteriosis 787:804 arg1 incidence 774:782 the incidence 770:782 the incidence of campylobacteriosis 770:804 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 0 28 theme jejuni 105:110 arg1 Pathogenicity 112:124 Campylobactor jejuni Pathogenicity 91:124 Campylobactor jejuni Pathogenicity 91:124 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 5 29 theme mucin 1009:1013 arg1 O-glycosylation 1015:1029 mucin O-glycosylation 1009:1029 mucin O-glycosylation 1009:1029 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 7 30 theme O-glycans 1260:1268 arg1 analysis 1248:1255 a detailed structural analysis 1226:1255 a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum 1226:1356 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 3 31 from abundant 418:425 arg1 crypts 441:446 the caecal crypts 430:446 the caecal crypts 430:446 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 5 32 theme intestinal 834:843 arg1 mucin 845:849 purified chicken intestinal mucin 817:849 purified chicken intestinal mucin 817:849 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 6 33 from regions 1054:1060 arg1 Mucins 1032:1037 Mucins 1032:1037 Mucins from different regions of the chicken intestine 1032:1085 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 10 34 theme present 1750:1756 arg1 structures 1739:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures 1683:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes 1683:1779 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 4 35 theme chicken 585:591 arg1 tissues 604:610 chicken intestinal tissues 585:610 chicken intestinal tissues 585:610 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 10 36 theme -like 1766:1770 arg1 epitopes 1772:1779 Sd(a)-like epitopes 1761:1779 Sd(a)-like epitopes 1761:1779 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 0 37 theme O-glycan 18:25 arg1 Structures 27:36 O-glycan Structures 18:36 O-glycan Structures from Chicken Intestinal Mucins 18:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 6 38 theme jejuni 1100:1105 arg1 binding 1107:1113 C. jejuni binding 1097:1113 C. jejuni binding 1097:1113 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 9 39 contain have 1575:1578 arg2 affinity 1595:1602 a high binding affinity 1580:1602 a high binding affinity to C. jejuni 1580:1615 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 9 39 contain have 1575:1578 arg1 residues 1559:1566 alpha 1-2 linked fucose residues 1535:1566 alpha 1-2 linked fucose residues 1535:1566 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 11 40 theme candidate 2022:2030 arg1 molecules 2032:2040 prospective candidate molecules 2010:2040 prospective candidate molecules aimed at reducing the incidence of infection 2010:2085 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 5 41 theme adenocarcinoma 934:947 arg1 line 954:957 the human colorectal adenocarcinoma cell line HCT-8 913:963 the human colorectal adenocarcinoma cell line HCT-8 913:963 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 3 42 theme cross-contamination 507:525 arg1 products 461:468 poultry products 453:468 poultry products 453:468 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 3 42 theme cross-contamination 507:525 arg1 result 497:502 a result 495:502 a result of cross-contamination during processing 495:543 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 0 43 theme Intestinal 51:60 arg1 Mucins 62:67 Chicken Intestinal Mucins 43:67 Chicken Intestinal Mucins 43:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 8 44 theme intestinal 1502:1511 arg1 tract 1513:1517 the chicken intestinal tract 1490:1517 the chicken intestinal tract 1490:1517 The O-glycans identified were abundantly sulfated compared with the human intestines, and sulfate moieties were present throughout the chicken intestinal tract. 25776888 11 45 theme adherence 1948:1956 arg1 properties 1971:1980 adherence and invasion properties 1948:1980 adherence and invasion properties of C. jejuni 1948:1993 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 4 46 dep identifying 702:712 arg1 provide 734:740 provide 734:740 identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis 702:804 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 47 theme molecular 638:646 arg1 mechanisms 648:657 the pathogenic molecular mechanisms 623:657 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 47 theme molecular 638:646 arg1 unknown 689:695 unknown 689:695 unknown 689:695 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 6 48 theme intestine>small 1162:1176 arg1 intestine>caecum 1178:1193 large intestine>small intestine>caecum 1156:1193 large intestine>small intestine>caecum 1156:1193 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 11 49 theme jejuni 1988:1993 arg1 properties 1971:1980 adherence and invasion properties 1948:1980 adherence and invasion properties of C. jejuni 1948:1993 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 1 50 theme diarrheal 269:277 arg1 illness 279:285 acute diarrheal illness 263:285 acute diarrheal illness 263:285 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 50 theme diarrheal 269:277 arg1 cause 300:304 a leading cause 290:304 a leading cause of mortality among children in developing countries 290:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 7 51 theme small 1330:1334 arg1 intestine 1336:1344 small intestine 1330:1344 small intestine 1330:1344 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 4 52 from mechanisms 648:657 arg1 humans 678:683 humans 678:683 humans 678:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 53 from interactions 550:561 arg1 humans 678:683 humans 678:683 humans 678:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 10 54 theme intestine 1806:1814 arg1 samples 1816:1822 large intestine samples 1800:1822 large intestine samples but not small intestine or caecum 1800:1856 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 1 55 theme leading 292:298 arg1 illness 279:285 acute diarrheal illness 263:285 acute diarrheal illness 263:285 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 55 theme leading 292:298 arg1 cause 300:304 a leading cause 290:304 a leading cause of mortality among children in developing countries 290:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 2 56 from commensal 373:381 arg1 chickens 386:393 chickens 386:393 chickens 386:393 C. jejuni are commensal in chickens. 25776888 11 57 theme invasion 1962:1969 arg1 properties 1971:1980 adherence and invasion properties 1948:1980 adherence and invasion properties of C. jejuni 1948:1993 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 10 58 theme small 1832:1836 arg1 intestine 1838:1846 small intestine 1832:1846 small intestine 1832:1846 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 1 59 theme food 215:218 arg1 poisoning 220:228 food poisoning 215:228 food poisoning in industrialized countries 215:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 60 theme responsible 199:209 arg1 jejuni 168:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 60 theme responsible 199:209 arg1 bacteria 190:197 the primary bacteria 178:197 the primary bacteria responsible for food poisoning in industrialized countries 178:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 5 61 theme human 917:921 arg1 line 954:957 the human colorectal adenocarcinoma cell line HCT-8 913:963 the human colorectal adenocarcinoma cell line HCT-8 913:963 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 11 62 theme intestinal 1907:1916 arg1 mucins 1918:1923 chicken intestinal mucins 1899:1923 chicken intestinal mucins 1899:1923 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 9 63 theme linked 1545:1550 arg1 residues 1559:1566 alpha 1-2 linked fucose residues 1535:1566 alpha 1-2 linked fucose residues 1535:1566 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 10 64 theme containing 1728:1737 arg1 structures 1739:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures 1683:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes 1683:1779 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 5 65 theme C. 900:901 arg1 jejuni 903:908 C. jejuni 900:908 C. jejuni 900:908 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 10 66 theme N-glycolylneuraminic/N-acetylneuraminic 1683:1721 arg1 structures 1739:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures 1683:1748 N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes 1683:1779 Additionally, N-glycolylneuraminic/N-acetylneuraminic acid containing structures present as Sd(a)-like epitopes were identified in large intestine samples but not small intestine or caecum. 25776888 1 67 theme Gram-negative 131:143 arg1 bacteria 145:152 Gram-negative bacteria 131:152 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 68 theme Campylobactor 154:166 arg1 jejuni 168:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 68 theme Campylobactor 154:166 arg1 bacteria 190:197 the primary bacteria 178:197 the primary bacteria responsible for food poisoning in industrialized countries 178:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 6 69 theme intestine 1077:1085 arg1 regions 1054:1060 different regions 1044:1060 different regions of the chicken intestine 1044:1085 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 9 70 theme alpha 1535:1539 arg1 residues 1559:1566 alpha 1-2 linked fucose residues 1535:1566 alpha 1-2 linked fucose residues 1535:1566 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 4 71 from colonization 662:673 arg1 humans 678:683 humans 678:683 humans 678:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 11 72 theme infection 2077:2085 arg1 incidence 2064:2072 the incidence 2060:2072 the incidence of infection 2060:2085 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 1 73 from poisoning 220:228 arg1 countries 248:256 industrialized countries 233:256 industrialized countries 233:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 9 74 theme binding 1587:1593 arg1 affinity 1595:1602 a high binding affinity 1580:1602 a high binding affinity to C. jejuni 1580:1615 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 0 75 theme Campylobactor 91:103 arg1 Pathogenicity 112:124 Campylobactor jejuni Pathogenicity 91:124 Campylobactor jejuni Pathogenicity 91:124 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 5 76 theme purified 817:824 arg1 mucin 845:849 purified chicken intestinal mucin 817:849 purified chicken intestinal mucin 817:849 Recently, purified chicken intestinal mucin was shown to attenuate adherence and invasion of C. jejuni in the human colorectal adenocarcinoma cell line HCT-8 in vitro, and this effect was attributed to mucin O-glycosylation. 25776888 1 77 theme primary 182:188 arg1 jejuni 168:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 77 theme primary 182:188 arg1 bacteria 190:197 the primary bacteria 178:197 the primary bacteria responsible for food poisoning in industrialized countries 178:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 7 78 dep detailed 1228:1235 arg1 structural 1237:1246 structural 1237:1246 structural 1237:1246 Here, we use LC-MS to perform a detailed structural analysis of O-glycans released from mucins purified from chicken large intestine, small intestine, and caecum. 25776888 11 79 theme structural 1868:1877 arg1 characterization 1879:1894 O-glycan structural characterization 1859:1894 O-glycan structural characterization of chicken intestinal mucins 1859:1923 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 9 80 link linked 1545:1550 arg1 residues 1559:1566 alpha 1-2 linked fucose residues 1535:1566 alpha 1-2 linked fucose residues 1535:1566 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 0 81 from Mucins 62:67 arg1 Identification 0:13 Identification 0:13 Identification of O-glycan Structures from Chicken Intestinal Mucins 0:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 0 81 from Mucins 62:67 arg1 Structures 27:36 O-glycan Structures 18:36 O-glycan Structures from Chicken Intestinal Mucins 18:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 9 82 theme fucose 1552:1557 arg1 residues 1559:1566 alpha 1-2 linked fucose residues 1535:1566 alpha 1-2 linked fucose residues 1535:1566 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 8 83 theme human 1427:1431 arg1 intestines 1433:1442 the human intestines 1423:1442 the human intestines 1423:1442 The O-glycans identified were abundantly sulfated compared with the human intestines, and sulfate moieties were present throughout the chicken intestinal tract. 25776888 11 84 theme prospective 2010:2020 arg1 molecules 2032:2040 prospective candidate molecules 2010:2040 prospective candidate molecules aimed at reducing the incidence of infection 2010:2085 O-glycan structural characterization of chicken intestinal mucins provides insights into adherence and invasion properties of C. jejuni, and may offer prospective candidate molecules aimed at reducing the incidence of infection. 25776888 4 85 theme intestinal 593:602 arg1 tissues 604:610 chicken intestinal tissues 585:610 chicken intestinal tissues 585:610 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 3 86 theme poultry 453:459 arg1 products 461:468 poultry products 453:468 poultry products 453:468 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 3 86 theme poultry 453:459 arg1 result 497:502 a result 495:502 a result of cross-contamination during processing 495:543 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 0 87 theme Structures 27:36 arg1 Identification 0:13 Identification 0:13 Identification of O-glycan Structures from Chicken Intestinal Mucins 0:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 1 88 theme mortality 309:317 arg1 cause 300:304 a leading cause 290:304 a leading cause of mortality among children in developing countries 290:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 88 theme mortality 309:317 arg1 jejuni 168:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni 127:173 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 88 theme mortality 309:317 arg1 illness 279:285 acute diarrheal illness 263:285 acute diarrheal illness 263:285 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 88 theme mortality 309:317 arg1 bacteria 190:197 the primary bacteria 178:197 the primary bacteria responsible for food poisoning in industrialized countries 178:256 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 9 89 theme small 1641:1645 arg1 intestines 1657:1666 the small and large intestines 1637:1666 the small and large intestines 1637:1666 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 8 90 theme sulfate 1449:1455 arg1 moieties 1457:1464 sulfate moieties 1449:1464 sulfate moieties 1449:1464 The O-glycans identified were abundantly sulfated compared with the human intestines, and sulfate moieties were present throughout the chicken intestinal tract. 25776888 0 91 theme Chicken 43:49 arg1 Mucins 62:67 Chicken Intestinal Mucins 43:67 Chicken Intestinal Mucins 43:67 Identification of O-glycan Structures from Chicken Intestinal Mucins Provides Insight into Campylobactor jejuni Pathogenicity. 25776888 9 92 theme large 1651:1655 arg1 intestines 1657:1666 the small and large intestines 1637:1666 the small and large intestines 1637:1666 Interestingly, alpha 1-2 linked fucose residues, which have a high binding affinity to C. jejuni, were identified in the small and large intestines. 25776888 6 93 theme C. 1097:1098 arg1 binding 1107:1113 C. jejuni binding 1097:1113 C. jejuni binding 1097:1113 Mucins from different regions of the chicken intestine inhibited C. jejuni binding and internalization differentially, with large intestine>small intestine>caecum. 25776888 4 94 theme C. 571:572 arg1 jejuni 574:579 C. jejuni 571:579 C. jejuni 571:579 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 1 95 theme developing 337:346 arg1 countries 348:356 developing countries 337:356 developing countries 337:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 3 96 theme caecal 434:439 arg1 crypts 441:446 the caecal crypts 430:446 the caecal crypts 430:446 They are particularly abundant in the caecal crypts, and poultry products are commonly infected as a result of cross-contamination during processing. 25776888 8 97 theme chicken 1494:1500 arg1 tract 1513:1517 the chicken intestinal tract 1490:1517 the chicken intestinal tract 1490:1517 The O-glycans identified were abundantly sulfated compared with the human intestines, and sulfate moieties were present throughout the chicken intestinal tract. 25776888 2 98 theme C. 359:360 arg1 commensal 373:381 commensal 373:381 commensal in chickens 373:393 C. jejuni are commensal in chickens. 25776888 2 98 theme C. 359:360 arg1 jejuni 362:367 C. jejuni 359:367 C. jejuni 359:367 C. jejuni are commensal in chickens. 25776888 1 99 theme acute 263:267 arg1 illness 279:285 acute diarrheal illness 263:285 acute diarrheal illness 263:285 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 1 99 theme acute 263:267 arg1 cause 300:304 a leading cause 290:304 a leading cause of mortality among children in developing countries 290:356 The Gram-negative bacteria Campylobactor jejuni is the primary bacteria responsible for food poisoning in industrialized countries, and acute diarrheal illness is a leading cause of mortality among children in developing countries. 25776888 4 100 theme colonization 662:673 arg1 mechanisms 648:657 the pathogenic molecular mechanisms 623:657 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 100 theme colonization 662:673 arg1 unknown 689:695 unknown 689:695 unknown 689:695 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 25776888 4 100 theme colonization 662:673 arg1 interactions 550:561 The interactions 546:561 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans 546:683 The interactions between C. jejuni and chicken intestinal tissues as well as the pathogenic molecular mechanisms of colonization in humans are unknown, but identifying these factors could provide potential targets to reduce the incidence of campylobacteriosis. 29404022 6 0 theme MS 951:952 arg1 instrument 954:963 MS instrument software and expandable 951:987 instrument 954:963 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 9 1 theme differentiation 1321:1335 arg1 degrees 1310:1316 different degrees 1300:1316 different degrees of differentiation 1300:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 9 2 theme present 1230:1236 arg1 N-glycans 1220:1228 the N-glycans 1216:1228 the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1216:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 0 3 theme hypothetical 97:108 arg1 database 110:117 a hypothetical database 95:117 a hypothetical database 95:117 N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database. 29404022 5 4 located found 844:848 arg2 residues 835:842 the common sugar residues 818:842 the common sugar residues found in N-glycans 818:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 5 4 located found 844:848 arg1 N-glycans 853:861 N-glycans 853:861 N-glycans 853:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 6 5 dep instrument 954:963 arg1 software 965:972 software 965:972 software 965:972 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 11 6 theme novel 1509:1513 arg1 strategy 1515:1522 the novel strategy 1505:1522 the novel strategy developed 1505:1532 Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis. 29404022 10 7 theme differentiation 1475:1489 arg1 states 1491:1496 their differentiation states 1469:1496 their differentiation states 1469:1496 Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. 29404022 3 8 theme labeled 572:578 arg1 glycans 580:586 labeled glycans 572:586 labeled glycans 572:586 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 1 9 theme proteins 141:148 arg1 responsible 153:163 responsible 153:163 responsible 153:163 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 1 9 theme proteins 141:148 arg1 glycosylation 124:136 The glycosylation 120:136 The glycosylation of proteins 120:148 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 5 10 theme compositional 768:780 arg1 possibilities 782:794 all compositional possibilities 764:794 all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans 764:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 8 11 theme accurate 1110:1117 arg1 data 1137:1140 much more accurate and comprehensive data than had been previously reported 1100:1174 much more accurate and comprehensive data than had been previously reported 1100:1174 The results provided much more accurate and comprehensive data than had been previously reported. 29404022 9 12 theme membrane 1245:1252 arg1 glycoproteins 1254:1266 the membrane glycoproteins 1241:1266 the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1241:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 2 13 theme efficient 284:292 arg1 release 294:300 an efficient release 281:300 an efficient release 281:300 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 10 14 theme N-glycan 1365:1372 arg1 data 1374:1377 Accurate and comprehensive N-glycan data 1338:1377 Accurate and comprehensive N-glycan data from those cells 1338:1394 Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. 29404022 1 15 theme structural 175:184 arg1 roles 201:205 their structural and functional roles 169:205 their structural and functional roles in many cellular activities 169:233 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 9 16 attach present 1230:1236 arg2 N-glycans 1220:1228 the N-glycans 1216:1228 the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1216:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 9 16 attach present 1230:1236 arg1 glycoproteins 1254:1266 the membrane glycoproteins 1241:1266 the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1241:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 10 17 theme comprehensive 1351:1363 arg1 data 1374:1377 Accurate and comprehensive N-glycan data 1338:1377 Accurate and comprehensive N-glycan data from those cells 1338:1394 Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. 29404022 7 18 from glycoproteins 1033:1045 arg1 N-glycans 990:998 N-glycans 990:998 N-glycans from four different well-studied glycoproteins 990:1045 N-glycans from four different well-studied glycoproteins were analyzed by this strategy. 29404022 9 19 with glycoproteins 1254:1266 arg1 degrees 1310:1316 different degrees 1300:1316 different degrees of differentiation 1300:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 8 20 theme comprehensive 1123:1135 arg1 data 1137:1140 much more accurate and comprehensive data than had been previously reported 1100:1174 much more accurate and comprehensive data than had been previously reported 1100:1174 The results provided much more accurate and comprehensive data than had been previously reported. 29404022 2 21 theme chromatography-mass 323:341 arg1 analysis 352:359 liquid chromatography-mass spectral analysis 316:359 liquid chromatography-mass spectral analysis 316:359 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 1 22 theme functional 190:199 arg1 roles 201:205 their structural and functional roles 169:205 their structural and functional roles in many cellular activities 169:233 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 9 23 theme gastric 1271:1277 arg1 cells 1289:1293 gastric carcinoma cells 1271:1293 gastric carcinoma cells 1271:1293 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 3 24 gly deglycosylation 511:525 arg1 kit 492:494 a recently commercialized kit 466:494 a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans 466:586 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 2 25 theme database 393:400 arg1 use 370:372 the use 366:372 the use of a comprehensive database to analyze N-glycans 366:421 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 1 26 gly glycosylation 124:136 arg1 proteins 141:148 proteins 141:148 proteins 141:148 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 9 27 theme carcinoma 1279:1287 arg1 cells 1289:1293 gastric carcinoma cells 1271:1293 gastric carcinoma cells 1271:1293 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 3 28 theme glycans 580:586 arg1 cleanup 561:567 cleanup 561:567 cleanup 561:567 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 3 28 theme glycans 580:586 arg1 labeling 548:555 rapid labeling 542:555 rapid labeling 542:555 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 2 29 theme liquid 316:321 arg1 analysis 352:359 liquid chromatography-mass spectral analysis 316:359 liquid chromatography-mass spectral analysis 316:359 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 7 30 gly glycoproteins 1033:1045 arg1 glycoproteins 1033:1045 four different well-studied glycoproteins 1005:1045 four different well-studied glycoproteins 1005:1045 N-glycans from four different well-studied glycoproteins were analyzed by this strategy. 29404022 2 31 theme comprehensive 379:391 arg1 database 393:400 a comprehensive database 377:400 a comprehensive database to analyze N-glycans 377:421 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 9 32 theme cells 1289:1293 arg1 glycoproteins 1254:1266 the membrane glycoproteins 1241:1266 the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1241:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 1 33 theme many 210:213 arg1 activities 224:233 many cellular activities 210:233 many cellular activities 210:233 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 3 34 theme analytical 428:437 arg1 method 439:444 The analytical method 424:444 The analytical method described 424:454 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 4 35 theme fluorescence 663:674 arg1 separation 615:624 the separation 611:624 the separation 611:624 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 35 theme fluorescence 663:674 arg1 detection 676:684 fluorescence detection 663:684 fluorescence detection of N-glycans 663:697 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 36 theme N-glycans 689:697 arg1 separation 615:624 the separation 611:624 the separation 611:624 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 36 theme N-glycans 689:697 arg1 analysis 650:657 mass spectrometry (MS) analysis 627:657 mass spectrometry (MS) analysis 627:657 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 36 theme N-glycans 689:697 arg1 detection 676:684 fluorescence detection 663:684 fluorescence detection of N-glycans 663:697 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 1 37 theme cellular 215:222 arg1 activities 224:233 many cellular activities 210:233 many cellular activities 210:233 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 5 38 theme common 822:827 arg1 residues 835:842 the common sugar residues 818:842 the common sugar residues found in N-glycans 818:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 9 39 gly glycoproteins 1254:1266 arg1 glycoproteins 1254:1266 the membrane glycoproteins 1241:1266 the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation 1241:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 9 40 theme different 1300:1308 arg1 degrees 1310:1316 different degrees 1300:1316 different degrees of differentiation 1300:1335 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 3 41 theme commercialized 477:490 arg1 kit 492:494 a recently commercialized kit 466:494 a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans 466:586 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 5 42 theme sugar 829:833 arg1 residues 835:842 the common sugar residues 818:842 the common sugar residues found in N-glycans 818:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 0 43 theme commercial 46:55 arg1 kit 57:59 a commercial kit 44:59 a commercial kit 44:59 N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database. 29404022 5 44 theme hypothetical 702:713 arg1 database 715:722 A hypothetical database 700:722 A hypothetical database 700:722 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 0 45 gly glycoproteins 24:36 arg1 glycoproteins 24:36 glycoproteins 24:36 glycoproteins using a commercial kit 24:59 N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database. 29404022 6 46 with compatible 935:944 arg1 expandable 978:987 expandable 978:987 expandable 978:987 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 6 46 with compatible 935:944 arg1 instrument 954:963 MS instrument software and expandable 951:987 instrument 954:963 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 11 47 theme N-glycan 1597:1604 arg1 analysis 1606:1613 N-glycan analysis 1597:1613 N-glycan analysis 1597:1613 Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis. 29404022 6 48 theme initial 871:877 arg1 version 879:885 the initial version 867:885 the initial version 867:885 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 9 49 used used 1200:1203 arg2 strategy 1182:1189 This strategy 1177:1189 This strategy 1177:1189 This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. 29404022 7 50 theme different 1010:1018 arg1 glycoproteins 1033:1045 four different well-studied glycoproteins 1005:1045 four different well-studied glycoproteins 1005:1045 N-glycans from four different well-studied glycoproteins were analyzed by this strategy. 29404022 6 51 contain contains 901:908 arg1 database 892:899 this database 887:899 this database 887:899 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 6 51 contain contains 901:908 arg2 database 892:899 this database 887:899 this database 887:899 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 6 51 contain contains 901:908 arg1 version 879:885 the initial version 867:885 the initial version 867:885 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 6 51 contain contains 901:908 arg2 N-glycans 917:925 >8,700 N-glycans 910:925 >8,700 N-glycans 910:925 In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. 29404022 11 52 theme analysis 1606:1613 arg1 comprehensiveness 1576:1592 comprehensiveness 1576:1592 comprehensiveness 1576:1592 Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis. 29404022 11 52 theme analysis 1606:1613 arg1 efficiency 1561:1570 efficiency 1561:1570 efficiency 1561:1570 Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis. 29404022 11 52 theme analysis 1606:1613 arg1 accuracy 1551:1558 accuracy 1551:1558 accuracy 1551:1558 Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis. 29404022 4 53 theme spectrometry 632:643 arg1 separation 615:624 the separation 611:624 the separation 611:624 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 53 theme spectrometry 632:643 arg1 analysis 650:657 mass spectrometry (MS) analysis 627:657 mass spectrometry (MS) analysis 627:657 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 2 54 theme spectral 343:350 arg1 analysis 352:359 liquid chromatography-mass spectral analysis 316:359 liquid chromatography-mass spectral analysis 316:359 This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. 29404022 4 55 theme mass 627:630 arg1 spectrometry 632:643 mass spectrometry 627:643 mass spectrometry (MS) analysis 627:657 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 4 55 theme mass 627:630 arg1 MS 646:647 MS 646:647 MS 646:647 This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. 29404022 1 56 from roles 201:205 arg1 activities 224:233 many cellular activities 210:233 many cellular activities 210:233 The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. 29404022 3 57 theme rapid 542:546 arg1 labeling 548:555 rapid labeling 542:555 rapid labeling 542:555 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 3 58 theme quick 505:509 arg1 deglycosylation 511:525 quick deglycosylation 505:525 quick deglycosylation 505:525 The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. 29404022 5 59 theme N-glycans 799:807 arg1 possibilities 782:794 all compositional possibilities 764:794 all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans 764:861 A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. 29404022 10 60 from cells 1390:1394 arg1 data 1374:1377 Accurate and comprehensive N-glycan data 1338:1377 Accurate and comprehensive N-glycan data from those cells 1338:1394 Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. 29404022 7 61 theme well-studied 1020:1031 arg1 glycoproteins 1033:1045 four different well-studied glycoproteins 1005:1045 four different well-studied glycoproteins 1005:1045 N-glycans from four different well-studied glycoproteins were analyzed by this strategy. 29404022 10 62 theme Accurate 1338:1345 arg1 data 1374:1377 Accurate and comprehensive N-glycan data 1338:1377 Accurate and comprehensive N-glycan data from those cells 1338:1394 Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. 27209430 4 0 theme microarray 924:933 arg1 analysis 935:942 lectin microarray analysis 917:942 lectin microarray analysis 917:942 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 13 1 dep CONCLUSIONS 2092:2102 arg1 This 2104:2107 This 2104:2107 This 2104:2107 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 13 1 dep CONCLUSIONS 2092:2102 arg1 report 2122:2127 the first report 2112:2127 the first report of a glycoprotein biomarker 2112:2155 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 2 2 theme matrix 350:355 arg1 metalloproteinase-3 357:375 matrix metalloproteinase-3 350:375 matrix metalloproteinase-3 (MMP-3) 350:383 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 2 theme matrix 350:355 arg1 protein 331:337 C-reactive protein 320:337 C-reactive protein (CRP) 320:343 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 2 theme matrix 350:355 arg1 biomarkers 414:423 serologic biomarkers 404:423 serologic biomarkers 404:423 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 2 theme matrix 350:355 arg1 MMP-3 378:382 MMP-3 378:382 MMP-3 378:382 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 1 3 gly glycoproteins 189:201 arg1 glycoproteins 189:201 serum glycoproteins 183:201 serum glycoproteins that exhibit disease-associated glycosylation changes 183:255 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 3 4 theme antibody-overlay 562:577 arg1 technology 597:606 a recently developed antibody-overlay lectin microarray technology 541:606 a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker 541:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 13 5 theme biomarker 2147:2155 arg1 This 2104:2107 This 2104:2107 This 2104:2107 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 13 5 theme biomarker 2147:2155 arg1 report 2122:2127 the first report 2112:2127 the first report of a glycoprotein biomarker 2112:2155 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 10 6 theme DAI 1746:1748 arg1 changes 1750:1756 DAI changes 1746:1756 DAI changes 1746:1756 Further validation using another cohort confirmed that this index correlated well with several DAIs and their components, and reflected DAI changes following RA treatment, with correlations greater than those for MMP-3 and CRP. 27209430 1 7 theme serum 183:187 arg1 glycoproteins 189:201 serum glycoproteins 183:201 serum glycoproteins that exhibit disease-associated glycosylation changes 183:255 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 7 8 theme MMP-3 1421:1425 arg1 levels 1405:1410 subnanogram levels 1393:1410 subnanogram levels of serum MMP-3 1393:1425 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 10 9 theme greater 1800:1806 arg1 correlations 1787:1798 correlations 1787:1798 correlations greater than those for MMP-3 and CRP 1787:1835 Further validation using another cohort confirmed that this index correlated well with several DAIs and their components, and reflected DAI changes following RA treatment, with correlations greater than those for MMP-3 and CRP. 27209430 3 10 theme microarray 586:595 arg1 technology 597:606 a recently developed antibody-overlay lectin microarray technology 541:606 a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker 541:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 1 11 theme disease-associated 216:233 arg1 changes 249:255 disease-associated glycosylation changes 216:255 disease-associated glycosylation changes 216:255 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 7 12 theme subnanogram 1393:1403 arg1 levels 1405:1410 subnanogram levels 1393:1410 subnanogram levels of serum MMP-3 1393:1425 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 13 13 theme first 2116:2120 arg1 This 2104:2107 This 2104:2107 This 2104:2107 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 13 13 theme first 2116:2120 arg1 report 2122:2127 the first report 2112:2127 the first report of a glycoprotein biomarker 2112:2155 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 3 14 theme quantitative 642:653 arg1 analysis 655:662 semicomprehensive and quantitative analysis 620:662 semicomprehensive and quantitative analysis of specific protein glycosylation 620:696 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 10 15 theme Further 1610:1616 arg1 validation 1618:1627 Further validation 1610:1627 Further validation using another cohort 1610:1648 Further validation using another cohort confirmed that this index correlated well with several DAIs and their components, and reflected DAI changes following RA treatment, with correlations greater than those for MMP-3 and CRP. 27209430 3 16 theme semicomprehensive 620:636 arg1 analysis 655:662 semicomprehensive and quantitative analysis 620:662 semicomprehensive and quantitative analysis of specific protein glycosylation 620:696 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 2 17 theme rheumatoid 293:302 arg1 RA 315:316 RA 315:316 RA 315:316 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 17 theme rheumatoid 293:302 arg1 arthritis 304:312 rheumatoid arthritis 293:312 rheumatoid arthritis (RA) 293:317 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 12 18 theme O-glycan 2064:2071 arg1 α-2,6-sialylation 2073:2089 O-glycan α-2,6-sialylation 2064:2089 O-glycan α-2,6-sialylation 2064:2089 Sialidase digestion revealed that the difference in quality was derived from O-glycan α-2,6-sialylation. 27209430 14 19 theme useful 2314:2319 arg1 Differences 2246:2256 Differences 2246:2256 Differences in the degree of serum MMP-3 α-2,6-sialylation 2246:2303 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 14 19 theme useful 2314:2319 arg1 index 2321:2325 a useful index 2312:2325 a useful index for estimating disease activity 2312:2357 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 1 20 gly glycosylated 165:176 arg1 BACKGROUND 121:130 BACKGROUND 121:130 BACKGROUND Nearly all secreted proteins are glycosylated 121:176 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 1 20 gly glycosylated 165:176 arg1 proteins 152:159 Nearly all secreted proteins 132:159 Nearly all secreted proteins 132:159 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 10 21 theme RA 1768:1769 arg1 treatment 1771:1779 RA treatment 1768:1779 RA treatment 1768:1779 Further validation using another cohort confirmed that this index correlated well with several DAIs and their components, and reflected DAI changes following RA treatment, with correlations greater than those for MMP-3 and CRP. 27209430 1 22 theme secreted 143:150 arg1 proteins 152:159 Nearly all secreted proteins 132:159 Nearly all secreted proteins 132:159 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 6 23 theme Synovial 1058:1065 arg1 MMP-3 1073:1077 Synovial fluid MMP-3 1058:1077 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) 1058:1135 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 4 24 theme METHODS 752:758 arg1 Serum 760:764 METHODS Serum 752:764 METHODS Serum 752:764 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 12 25 from difference 2025:2034 arg1 quality 2039:2045 quality 2039:2045 quality 2039:2045 Sialidase digestion revealed that the difference in quality was derived from O-glycan α-2,6-sialylation. 27209430 5 26 theme disease 947:953 arg1 DAI 971:973 DAI 971:973 DAI 971:973 A disease activity index (DAI) based on lectin signal was developed and validated using another cohort (n = 60). 27209430 5 26 theme disease 947:953 arg1 index 964:968 A disease activity index 945:968 A disease activity index (DAI) based on lectin signal 945:997 A disease activity index (DAI) based on lectin signal was developed and validated using another cohort (n = 60). 27209430 2 27 used used 396:399 arg2 MMP-3 378:382 MMP-3 378:382 MMP-3 378:382 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 27 used used 396:399 arg2 CRP 340:342 CRP 340:342 CRP 340:342 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 27 used used 396:399 arg2 metalloproteinase-3 357:375 matrix metalloproteinase-3 350:375 matrix metalloproteinase-3 (MMP-3) 350:383 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 27 used used 396:399 arg2 protein 331:337 C-reactive protein 320:337 C-reactive protein (CRP) 320:343 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 27 used used 396:399 arg2 biomarkers 414:423 serologic biomarkers 404:423 serologic biomarkers 404:423 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 14 28 theme α-2,6-sialylation 2287:2303 arg1 degree 2265:2270 the degree 2261:2270 the degree of serum MMP-3 α-2,6-sialylation 2261:2303 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 0 29 theme disease 79:85 arg1 activity 87:94 disease activity 79:94 disease activity of rheumatoid arthritis 79:118 Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis. 27209430 3 30 theme protein 676:682 arg1 glycosylation 684:696 specific protein glycosylation 667:696 specific protein glycosylation 667:696 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 4 31 theme disease 813:819 arg1 activity 821:828 activity 821:828 activity 821:828 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 14 32 theme serum 2275:2279 arg1 α-2,6-sialylation 2287:2303 serum MMP-3 α-2,6-sialylation 2275:2303 serum MMP-3 α-2,6-sialylation 2275:2303 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 7 33 theme Agrocybe 1227:1234 arg1 galectin 1248:1255 Agrocybe cylindracea galectin [ACG 1227:1260 Agrocybe cylindracea galectin [ACG 1227:1260 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 33 theme Agrocybe 1227:1234 arg1 lectin 1219:1224 a sialic acid-binding lectin 1197:1224 a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) 1197:1262 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 34 dep Jacalin 1294:1300 arg1 [ACA 1374:1377 [ACA 1374:1377 [ACA 1374:1377 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 13 35 from activity 2213:2220 arg1 diseases 2236:2243 autoimmune diseases 2225:2243 autoimmune diseases 2225:2243 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 1 36 dep BACKGROUND 121:130 arg1 glycosylated 165:176 glycosylated 165:176 are glycosylated 161:176 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 0 37 theme arthritis 110:118 arg1 activity 87:94 disease activity 79:94 disease activity of rheumatoid arthritis 79:118 Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis. 27209430 7 38 theme caudatus 1354:1361 arg1 agglutinin 1363:1372 Amaranthus caudatus agglutinin 1343:1372 Amaranthus caudatus agglutinin 1343:1372 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 38 theme caudatus 1354:1361 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 2 39 theme sufficient 440:449 arg1 specificity 451:461 specificity 451:461 specificity 451:461 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 11 40 theme ACG/Jacalin 1881:1891 arg1 score 1893:1897 a high ACG/Jacalin score 1874:1897 a high ACG/Jacalin score 1874:1897 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 27209430 7 41 theme sialic 1199:1204 arg1 galectin 1248:1255 Agrocybe cylindracea galectin [ACG 1227:1260 Agrocybe cylindracea galectin [ACG 1227:1260 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 41 theme sialic 1199:1204 arg1 lectin 1219:1224 a sialic acid-binding lectin 1197:1224 a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) 1197:1262 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 5 42 theme lectin 985:990 arg1 signal 992:997 lectin signal 985:997 lectin signal 985:997 A disease activity index (DAI) based on lectin signal was developed and validated using another cohort (n = 60). 27209430 0 43 theme matrix 14:19 arg1 metalloproteinase-3 21:39 matrix metalloproteinase-3 14:39 matrix metalloproteinase-3 14:39 Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis. 27209430 7 44 dep lectins 1285:1291 arg1 Jacalin 1294:1300 Jacalin 1294:1300 Jacalin 1294:1300 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 44 dep lectins 1285:1291 arg1 agglutinin 1363:1372 Amaranthus caudatus agglutinin 1343:1372 Amaranthus caudatus agglutinin 1343:1372 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 44 dep lectins 1285:1291 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 44 dep lectins 1285:1291 arg1 agglutinin 1321:1330 Agaricus bisporus agglutinin 1303:1330 Agaricus bisporus agglutinin [ABA] 1303:1336 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 14 45 from Differences 2246:2256 arg1 degree 2265:2270 the degree 2261:2270 the degree of serum MMP-3 α-2,6-sialylation 2261:2303 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 2 46 theme serologic 404:412 arg1 metalloproteinase-3 357:375 matrix metalloproteinase-3 350:375 matrix metalloproteinase-3 (MMP-3) 350:383 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 46 theme serologic 404:412 arg1 protein 331:337 C-reactive protein 320:337 C-reactive protein (CRP) 320:343 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 46 theme serologic 404:412 arg1 biomarkers 414:423 serologic biomarkers 404:423 serologic biomarkers 404:423 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 9 47 theme disease 1592:1598 arg1 activity 1600:1607 disease activity 1592:1607 disease activity 1592:1607 The resultant index, ACG/Jacalin, correlated well with disease activity. 27209430 3 48 theme comparative 490:500 arg1 profiling 516:524 comparative glycosylation profiling 490:524 comparative glycosylation profiling of MMP-3 490:533 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 7 49 theme Intense 1164:1170 arg1 signals 1172:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 50 theme Agaricus 1303:1310 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 50 theme Agaricus 1303:1310 arg1 agglutinin 1321:1330 Agaricus bisporus agglutinin 1303:1330 Agaricus bisporus agglutinin [ABA] 1303:1336 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 51 dep agglutinin 1321:1330 arg1 [ABA 1332:1335 [ABA 1332:1335 [ABA 1332:1335 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 3 52 theme disease 724:730 arg1 biomarker 741:749 an RA-specific disease activity biomarker 709:749 an RA-specific disease activity biomarker 709:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 3 53 theme MMP-3 529:533 arg1 profiling 516:524 comparative glycosylation profiling 490:524 comparative glycosylation profiling of MMP-3 490:533 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 12 54 theme Sialidase 1987:1995 arg1 digestion 1997:2005 Sialidase digestion 1987:2005 Sialidase digestion 1987:2005 Sialidase digestion revealed that the difference in quality was derived from O-glycan α-2,6-sialylation. 27209430 4 55 with patients 781:788 arg1 n = 24 799:804 n = 24 799:804 n = 24 799:804 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 4 55 with patients 781:788 arg1 RA 795:796 RA 795:796 RA (n = 24) 795:805 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 4 56 theme lectin 917:922 arg1 analysis 935:942 lectin microarray analysis 917:942 lectin microarray analysis 917:942 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 14 57 theme disease 2342:2348 arg1 activity 2350:2357 disease activity 2342:2357 disease activity 2342:2357 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 3 58 theme developed 552:560 arg1 technology 597:606 a recently developed antibody-overlay lectin microarray technology 541:606 a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker 541:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 13 59 theme glycoprotein 2134:2145 arg1 biomarker 2147:2155 a glycoprotein biomarker 2132:2155 a glycoprotein biomarker 2132:2155 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 6 60 from MMP-3 1073:1077 arg1 patients 1082:1089 patients 1082:1089 patients with RA 1082:1097 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 6 60 from MMP-3 1073:1077 arg1 patients 1103:1110 patients 1103:1110 patients with osteoarthritis (OA) 1103:1135 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 7 61 located observed 1185:1192 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg1 agglutinin 1321:1330 Agaricus bisporus agglutinin 1303:1330 Agaricus bisporus agglutinin [ABA] 1303:1336 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg2 signals 1172:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg1 galectin 1248:1255 Agrocybe cylindracea galectin [ACG 1227:1260 Agrocybe cylindracea galectin [ACG 1227:1260 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg1 agglutinin 1363:1372 Amaranthus caudatus agglutinin 1343:1372 Amaranthus caudatus agglutinin 1343:1372 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg1 Jacalin 1294:1300 Jacalin 1294:1300 Jacalin 1294:1300 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 61 located observed 1185:1192 arg1 lectin 1219:1224 a sialic acid-binding lectin 1197:1224 a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) 1197:1262 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 3 62 theme lectin 579:584 arg1 technology 597:606 a recently developed antibody-overlay lectin microarray technology 541:606 a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker 541:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 12 63 attach derived 2051:2057 arg1 α-2,6-sialylation 2073:2089 O-glycan α-2,6-sialylation 2064:2089 O-glycan α-2,6-sialylation 2064:2089 Sialidase digestion revealed that the difference in quality was derived from O-glycan α-2,6-sialylation. 27209430 12 63 attach derived 2051:2057 arg2 difference 2025:2034 the difference 2021:2034 the difference in quality 2021:2045 Sialidase digestion revealed that the difference in quality was derived from O-glycan α-2,6-sialylation. 27209430 7 64 theme serum 1415:1419 arg1 MMP-3 1421:1425 serum MMP-3 1415:1425 serum MMP-3 1415:1425 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 1 65 theme glycosylation 235:247 arg1 changes 249:255 disease-associated glycosylation changes 216:255 disease-associated glycosylation changes 216:255 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 1 66 contain have 257:260 arg1 glycoproteins 189:201 serum glycoproteins 183:201 serum glycoproteins that exhibit disease-associated glycosylation changes 183:255 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 1 66 contain have 257:260 arg2 potential 262:270 potential 262:270 potential 262:270 BACKGROUND Nearly all secreted proteins are glycosylated, and serum glycoproteins that exhibit disease-associated glycosylation changes have potential to be biomarkers. 27209430 11 67 with patients 1933:1940 arg1 RA 1947:1948 RA 1947:1948 RA 1947:1948 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 27209430 11 68 theme patients 1933:1940 arg1 fluid 1924:1928 synovial fluid 1915:1928 synovial fluid of patients with RA 1915:1948 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 27209430 8 69 theme MMP-3 1470:1474 arg1 quantity 1476:1483 MMP-3 quantity 1470:1483 MMP-3 quantity 1470:1483 ACG, ABA, and ACA revealed differences in MMP-3 quantity, and Jacalin revealed differences in MMP-3 quality. 27209430 3 70 theme specific 667:674 arg1 glycosylation 684:696 specific protein glycosylation 667:696 specific protein glycosylation 667:696 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 2 71 theme C-reactive 320:329 arg1 biomarkers 414:423 serologic biomarkers 404:423 serologic biomarkers 404:423 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 71 theme C-reactive 320:329 arg1 CRP 340:342 CRP 340:342 CRP 340:342 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 71 theme C-reactive 320:329 arg1 protein 331:337 C-reactive protein 320:337 C-reactive protein (CRP) 320:343 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 2 71 theme C-reactive 320:329 arg1 metalloproteinase-3 357:375 matrix metalloproteinase-3 350:375 matrix metalloproteinase-3 (MMP-3) 350:383 In rheumatoid arthritis (RA), C-reactive protein (CRP), and matrix metalloproteinase-3 (MMP-3) are widely used as serologic biomarkers, but they lack sufficient specificity or precision. 27209430 6 72 theme fluid 1067:1071 arg1 MMP-3 1073:1077 Synovial fluid MMP-3 1058:1077 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) 1058:1135 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 8 73 from differences 1455:1465 arg1 quantity 1476:1483 MMP-3 quantity 1470:1483 MMP-3 quantity 1470:1483 ACG, ABA, and ACA revealed differences in MMP-3 quantity, and Jacalin revealed differences in MMP-3 quality. 27209430 13 74 theme autoimmune 2225:2234 arg1 diseases 2236:2243 autoimmune diseases 2225:2243 autoimmune diseases 2225:2243 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 7 75 theme cylindracea 1236:1246 arg1 galectin 1248:1255 Agrocybe cylindracea galectin [ACG 1227:1260 Agrocybe cylindracea galectin [ACG 1227:1260 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 75 theme cylindracea 1236:1246 arg1 lectin 1219:1224 a sialic acid-binding lectin 1197:1224 a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) 1197:1262 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 5 76 theme activity 955:962 arg1 DAI 971:973 DAI 971:973 DAI 971:973 A disease activity index (DAI) based on lectin signal was developed and validated using another cohort (n = 60). 27209430 5 76 theme activity 955:962 arg1 index 964:968 A disease activity index 945:968 A disease activity index (DAI) based on lectin signal 945:997 A disease activity index (DAI) based on lectin signal was developed and validated using another cohort (n = 60). 27209430 14 77 theme MMP-3 2281:2285 arg1 α-2,6-sialylation 2287:2303 serum MMP-3 α-2,6-sialylation 2275:2303 serum MMP-3 α-2,6-sialylation 2275:2303 Differences in the degree of serum MMP-3 α-2,6-sialylation may be a useful index for estimating disease activity. 27209430 0 78 theme rheumatoid 99:108 arg1 arthritis 110:118 rheumatoid arthritis 99:118 rheumatoid arthritis 99:118 Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis. 27209430 8 79 theme MMP-3 1522:1526 arg1 quality 1528:1534 MMP-3 quality 1522:1534 MMP-3 quality 1522:1534 ACG, ABA, and ACA revealed differences in MMP-3 quantity, and Jacalin revealed differences in MMP-3 quality. 27209430 9 80 theme resultant 1541:1549 arg1 index 1551:1555 The resultant index 1537:1555 The resultant index 1537:1555 The resultant index, ACG/Jacalin, correlated well with disease activity. 27209430 9 80 theme resultant 1541:1549 arg1 ACG/Jacalin 1558:1568 ACG/Jacalin 1558:1568 ACG/Jacalin 1558:1568 The resultant index, ACG/Jacalin, correlated well with disease activity. 27209430 7 81 theme acid-binding 1206:1217 arg1 galectin 1248:1255 Agrocybe cylindracea galectin [ACG 1227:1260 Agrocybe cylindracea galectin [ACG 1227:1260 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 81 theme acid-binding 1206:1217 arg1 lectin 1219:1224 a sialic acid-binding lectin 1197:1224 a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) 1197:1262 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 82 theme Amaranthus 1343:1352 arg1 caudatus 1354:1361 Amaranthus caudatus 1343:1361 Amaranthus caudatus agglutinin 1343:1372 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 8 83 from differences 1507:1517 arg1 quality 1528:1534 MMP-3 quality 1522:1534 MMP-3 quality 1522:1534 ACG, ABA, and ACA revealed differences in MMP-3 quantity, and Jacalin revealed differences in MMP-3 quality. 27209430 0 84 theme metalloproteinase-3 21:39 arg1 Alteration 0:9 Alteration 0:9 Alteration of matrix metalloproteinase-3 0:39 Alteration of matrix metalloproteinase-3 O-glycan structure as a biomarker for disease activity of rheumatoid arthritis. 27209430 13 85 theme disease 2205:2211 arg1 activity 2213:2220 disease activity 2205:2220 disease activity in autoimmune diseases 2205:2243 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 11 86 with patients 1969:1976 arg1 OA 1983:1984 OA 1983:1984 OA 1983:1984 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 27209430 7 87 theme bisporus 1312:1319 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 87 theme bisporus 1312:1319 arg1 agglutinin 1321:1330 Agaricus bisporus agglutinin 1303:1330 Agaricus bisporus agglutinin [ABA] 1303:1336 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 3 88 theme glycosylation 684:696 arg1 analysis 655:662 semicomprehensive and quantitative analysis 620:662 semicomprehensive and quantitative analysis of specific protein glycosylation 620:696 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 11 89 theme synovial 1915:1922 arg1 fluid 1924:1928 synovial fluid 1915:1928 synovial fluid of patients with RA 1915:1948 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 27209430 7 90 theme RESULTS 1156:1162 arg1 signals 1172:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals 1156:1178 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 13 91 theme local 2182:2186 arg1 lesion 2188:2193 a local lesion 2180:2193 a local lesion to assess disease activity in autoimmune diseases 2180:2243 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 6 92 with patients 1103:1110 arg1 OA 1133:1134 OA 1133:1134 OA 1133:1134 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 6 92 with patients 1103:1110 arg1 osteoarthritis 1117:1130 osteoarthritis 1117:1130 osteoarthritis (OA) 1117:1135 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 6 92 with patients 1103:1110 arg1 RA 1096:1097 RA 1096:1097 RA 1096:1097 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 3 93 theme activity 732:739 arg1 biomarker 741:749 an RA-specific disease activity biomarker 709:749 an RA-specific disease activity biomarker 709:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 3 94 theme glycosylation 502:514 arg1 profiling 516:524 comparative glycosylation profiling 490:524 comparative glycosylation profiling of MMP-3 490:533 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 13 95 gly glycoprotein 2134:2145 arg1 glycoprotein 2134:2145 a glycoprotein biomarker 2132:2155 a glycoprotein biomarker 2132:2155 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 3 96 theme RA-specific 712:722 arg1 biomarker 741:749 an RA-specific disease activity biomarker 709:749 an RA-specific disease activity biomarker 709:749 We performed comparative glycosylation profiling of MMP-3 using a recently developed antibody-overlay lectin microarray technology that allows semicomprehensive and quantitative analysis of specific protein glycosylation to develop an RA-specific disease activity biomarker. 27209430 10 97 theme several 1697:1703 arg1 DAIs 1705:1708 several DAIs 1697:1708 several DAIs 1697:1708 Further validation using another cohort confirmed that this index correlated well with several DAIs and their components, and reflected DAI changes following RA treatment, with correlations greater than those for MMP-3 and CRP. 27209430 13 98 theme glycan 2163:2168 arg1 change 2170:2175 glycan change 2163:2175 glycan change 2163:2175 CONCLUSIONS This is the first report of a glycoprotein biomarker using glycan change at a local lesion to assess disease activity in autoimmune diseases. 27209430 6 99 with patients 1082:1089 arg1 OA 1133:1134 OA 1133:1134 OA 1133:1134 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 6 99 with patients 1082:1089 arg1 osteoarthritis 1117:1130 osteoarthritis 1117:1130 osteoarthritis (OA) 1117:1135 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 6 99 with patients 1082:1089 arg1 RA 1096:1097 RA 1096:1097 RA 1096:1097 Synovial fluid MMP-3 in patients with RA and patients with osteoarthritis (OA) was also analyzed. 27209430 7 100 theme O-glycan-binding 1268:1283 arg1 Jacalin 1294:1300 Jacalin 1294:1300 Jacalin 1294:1300 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 100 theme O-glycan-binding 1268:1283 arg1 agglutinin 1363:1372 Amaranthus caudatus agglutinin 1343:1372 Amaranthus caudatus agglutinin 1343:1372 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 100 theme O-glycan-binding 1268:1283 arg1 lectins 1285:1291 O-glycan-binding lectins 1268:1291 O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) 1268:1379 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 7 100 theme O-glycan-binding 1268:1283 arg1 agglutinin 1321:1330 Agaricus bisporus agglutinin 1303:1330 Agaricus bisporus agglutinin [ABA] 1303:1336 RESULTS Intense signals were observed on a sialic acid-binding lectin (Agrocybe cylindracea galectin [ACG]) and O-glycan-binding lectins (Jacalin, Agaricus bisporus agglutinin [ABA], and Amaranthus caudatus agglutinin [ACA]) by applying subnanogram levels of serum MMP-3. 27209430 4 101 theme composite 847:855 arg1 measures 857:864 composite measures 847:864 composite measures 847:864 METHODS Serum was taken from patients with RA (n = 24) whose disease activity was scored using composite measures, and MMP-3 was immunoprecipitated and subjected to lectin microarray analysis. 27209430 11 102 theme high 1876:1879 arg1 score 1893:1897 a high ACG/Jacalin score 1874:1897 a high ACG/Jacalin score 1874:1897 Furthermore, MMP-3, which generated a high ACG/Jacalin score, accumulated in synovial fluid of patients with RA but not in that of patients with OA. 24612801 10 0 theme important 1472:1480 arg1 role 1482:1485 an important role 1469:1485 an important role 1469:1485 In addition, heat-labile structures in κ-CN may play an important role in maintenance of κ-CN binding to HRV. 24612801 7 1 theme evanescent-field 967:982 arg1 assay 1006:1010 an evanescent-field fluorescence-assisted assay 964:1010 an evanescent-field fluorescence-assisted assay 964:1010 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 9 2 theme direct 1361:1366 arg1 binding 1368:1374 direct binding 1361:1374 direct binding to viral particles 1361:1393 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 0 3 theme direct 61:66 arg1 binding 68:74 direct binding 61:74 direct binding of glycans to HRV 61:92 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 4 4 gly glycosylated 556:567 arg1 protein 569:575 the only glycosylated protein 547:575 the only glycosylated protein that has been identified 547:600 Among the components of casein, κ-CN is the only glycosylated protein that has been identified. 24612801 4 4 gly glycosylated 556:567 arg1 κ-CN 539:542 κ-CN 539:542 κ-CN 539:542 Among the components of casein, κ-CN is the only glycosylated protein that has been identified. 24612801 8 5 theme heat 1257:1260 arg1 treatment 1262:1270 heat treatment 1257:1270 heat treatment 1257:1270 Although the heated κ-CN retained inhibitory activity in a neutralization assay, the activity was weaker than that observed before heat treatment. 24612801 0 6 theme glycans 79:85 arg1 binding 68:74 direct binding 61:74 direct binding of glycans to HRV 61:92 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 6 7 theme o-glycosidase 825:837 arg1 treatment 839:847 o-glycosidase treatment 825:847 o-glycosidase treatment 825:847 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 10 8 from structures 1441:1450 arg1 κ-CN 1455:1458 κ-CN 1455:1458 κ-CN 1455:1458 In addition, heat-labile structures in κ-CN may play an important role in maintenance of κ-CN binding to HRV. 24612801 10 9 theme binding 1510:1516 arg1 maintenance 1490:1500 maintenance 1490:1500 maintenance of κ-CN binding to HRV 1490:1523 In addition, heat-labile structures in κ-CN may play an important role in maintenance of κ-CN binding to HRV. 24612801 4 10 theme glycosylated 556:567 arg1 protein 569:575 the only glycosylated protein 547:575 the only glycosylated protein that has been identified 547:600 Among the components of casein, κ-CN is the only glycosylated protein that has been identified. 24612801 4 10 theme glycosylated 556:567 arg1 κ-CN 539:542 κ-CN 539:542 κ-CN 539:542 Among the components of casein, κ-CN is the only glycosylated protein that has been identified. 24612801 6 11 theme antiviral 925:933 arg1 activity 935:942 the antiviral activity 921:942 the antiviral activity of CN 921:948 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 10 12 theme κ-CN 1505:1508 arg1 binding 1510:1516 κ-CN binding 1505:1516 κ-CN binding to HRV 1505:1523 In addition, heat-labile structures in κ-CN may play an important role in maintenance of κ-CN binding to HRV. 24612801 6 13 theme Desialylated 706:717 arg1 CN 719:720 Desialylated CN 706:720 Desialylated CN obtained by neuraminidase treatment 706:756 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 5 14 theme glycan 642:647 arg1 residues 649:656 the glycan residues 638:656 the glycan residues in κ-CN 638:664 Therefore, we investigated whether the glycan residues in κ-CN were involved in the anti-HRV activity. 24612801 3 15 theme protective 398:407 arg1 effect 409:414 the protective effect 394:414 the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk 394:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 2 16 theme mature 224:229 arg1 milk 231:234 both human and bovine mature milk 202:234 both human and bovine mature milk 202:234 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 8 17 theme neutralization 1185:1198 arg1 assay 1200:1204 a neutralization assay 1183:1204 a neutralization assay 1183:1204 Although the heated κ-CN retained inhibitory activity in a neutralization assay, the activity was weaker than that observed before heat treatment. 24612801 1 18 theme major 122:126 arg1 agent 138:142 a major etiologic agent 120:142 a major etiologic agent of severe infantile gastroenteritis 120:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 1 18 theme major 122:126 arg1 rotavirus 101:109 Human rotavirus 95:109 Human rotavirus (HRV) 95:115 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 7 19 theme heated 1056:1061 arg1 casein 1063:1068 heated casein 1056:1068 heated casein (at 95°C for 30 min) 1056:1089 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 2 20 theme bovine 217:222 arg1 milk 231:234 both human and bovine mature milk 202:234 both human and bovine mature milk 202:234 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 1 21 theme etiologic 128:136 arg1 agent 138:142 a major etiologic agent 120:142 a major etiologic agent of severe infantile gastroenteritis 120:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 1 21 theme etiologic 128:136 arg1 rotavirus 101:109 Human rotavirus 95:109 Human rotavirus (HRV) 95:115 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 7 22 theme viral 1096:1100 arg1 manner 1118:1123 a viral titer-dependent manner 1094:1123 a viral titer-dependent manner 1094:1123 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 6 23 theme deglycosylated 795:808 arg1 CN 810:811 deglycosylated CN 795:811 deglycosylated CN obtained by o-glycosidase treatment 795:847 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 2 24 theme activity 312:319 arg1 mechanism 294:302 the mechanism 290:302 the mechanism of this activity 290:319 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 0 25 theme Bovine 0:5 arg1 κ-casein 7:14 Bovine κ-casein 0:14 Bovine κ-casein 0:14 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 0 26 theme human 25:29 arg1 HRV 42:44 HRV 42:44 HRV 42:44 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 0 26 theme human 25:29 arg1 rotavirus 31:39 human rotavirus 25:39 human rotavirus (HRV) infection 25:55 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 7 27 theme HRV 1024:1026 arg1 particles 1028:1036 HRV particles 1024:1036 HRV particles 1024:1036 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 1 28 theme severe 147:152 arg1 gastroenteritis 164:178 severe infantile gastroenteritis 147:178 severe infantile gastroenteritis 147:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 10 29 theme heat-labile 1429:1439 arg1 structures 1441:1450 heat-labile structures 1429:1450 heat-labile structures in κ-CN 1429:1458 In addition, heat-labile structures in κ-CN may play an important role in maintenance of κ-CN binding to HRV. 24612801 3 30 dep colostrum 449:457 arg1 d 464:464 6-7 d 460:464 6-7 d after parturition 460:482 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 1 31 theme infantile 154:162 arg1 gastroenteritis 164:178 severe infantile gastroenteritis 147:178 severe infantile gastroenteritis 147:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 3 32 theme κ-CN 426:429 arg1 effect 409:414 the protective effect 394:414 the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk 394:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 5 33 from residues 649:656 arg1 κ-CN 661:664 κ-CN 661:664 κ-CN 661:664 Therefore, we investigated whether the glycan residues in κ-CN were involved in the anti-HRV activity. 24612801 2 34 from milk 231:234 arg1 κ-CN 191:194 κ-CN 191:194 κ-CN 191:194 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 2 34 from milk 231:234 arg1 κ-Casein 181:188 κ-Casein 181:188 κ-Casein (κ-CN) from both human and bovine mature milk 181:234 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 1 35 theme gastroenteritis 164:178 arg1 agent 138:142 a major etiologic agent 120:142 a major etiologic agent of severe infantile gastroenteritis 120:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 1 35 theme gastroenteritis 164:178 arg1 rotavirus 101:109 Human rotavirus 95:109 Human rotavirus (HRV) 95:115 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 0 36 theme rotavirus 31:39 arg1 infection 47:55 human rotavirus (HRV) infection 25:55 human rotavirus (HRV) infection 25:55 Bovine κ-casein inhibits human rotavirus (HRV) infection via direct binding of glycans to HRV. 24612801 3 37 theme present 347:353 arg1 study 355:359 The present study 343:359 The present study 343:359 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 9 38 theme κ-CN 1335:1338 arg1 mechanism 1315:1323 the inhibitory mechanism 1300:1323 the inhibitory mechanism of bovine κ-CN against HRV 1300:1350 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 6 39 gly Desialylated 706:717 arg1 CN 719:720 Desialylated CN 706:720 Desialylated CN obtained by neuraminidase treatment 706:756 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 5 40 theme anti-HRV 687:694 arg1 activity 696:703 the anti-HRV activity 683:703 the anti-HRV activity 683:703 Therefore, we investigated whether the glycan residues in κ-CN were involved in the anti-HRV activity. 24612801 9 41 theme viral 1379:1383 arg1 particles 1385:1393 viral particles 1379:1393 viral particles 1379:1393 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 7 42 theme fluorescence-assisted 984:1004 arg1 assay 1006:1010 an evanescent-field fluorescence-assisted assay 964:1010 an evanescent-field fluorescence-assisted assay 964:1010 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 6 43 theme anti-HRV 768:775 arg1 activity 777:784 anti-HRV activity 768:784 anti-HRV activity 768:784 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 3 44 theme mature 494:499 arg1 milk 501:504 mature milk 494:504 mature milk 494:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 7 45 from 95°C 1074:1077 arg1 casein 1063:1068 heated casein 1056:1068 heated casein (at 95°C for 30 min) 1056:1089 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 3 46 theme bovine 419:424 arg1 κ-CN 426:429 bovine κ-CN 419:429 bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk 419:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 9 47 theme inhibitory 1304:1313 arg1 mechanism 1315:1323 the inhibitory mechanism 1300:1323 the inhibitory mechanism of bovine κ-CN against HRV 1300:1350 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 9 48 theme bovine 1328:1333 arg1 κ-CN 1335:1338 bovine κ-CN 1328:1338 bovine κ-CN 1328:1338 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 1 49 theme Human 95:99 arg1 rotavirus 101:109 Human rotavirus 95:109 Human rotavirus (HRV) 95:115 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 1 49 theme Human 95:99 arg1 agent 138:142 a major etiologic agent 120:142 a major etiologic agent of severe infantile gastroenteritis 120:178 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 1 49 theme Human 95:99 arg1 HRV 112:114 HRV 112:114 HRV 112:114 Human rotavirus (HRV) is a major etiologic agent of severe infantile gastroenteritis. 24612801 2 50 contain have 257:260 arg1 κ-CN 191:194 κ-CN 191:194 κ-CN 191:194 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 2 50 contain have 257:260 arg1 κ-Casein 181:188 κ-Casein 181:188 κ-Casein (κ-CN) from both human and bovine mature milk 181:234 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 2 50 contain have 257:260 arg2 activity 271:278 anti-HRV activity 262:278 anti-HRV activity 262:278 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 3 51 attach derived 431:437 arg2 κ-CN 426:429 bovine κ-CN 419:429 bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk 419:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 3 51 attach derived 431:437 arg1 milk 501:504 mature milk 494:504 mature milk 494:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 3 51 attach derived 431:437 arg1 colostrum 449:457 late colostrum 444:457 late colostrum (6-7 d after parturition) 444:483 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 3 52 theme 6-7 460:462 arg1 d 464:464 6-7 d 460:464 6-7 d after parturition 460:482 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 6 53 theme neuraminidase 734:746 arg1 treatment 748:756 neuraminidase treatment 734:756 neuraminidase treatment 734:756 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 2 54 theme human 207:211 arg1 milk 231:234 both human and bovine mature milk 202:234 both human and bovine mature milk 202:234 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 2 55 theme anti-HRV 262:269 arg1 activity 271:278 anti-HRV activity 262:278 anti-HRV activity 262:278 κ-Casein (κ-CN) from both human and bovine mature milk has been reported to have anti-HRV activity; however, the mechanism of this activity is poorly understood. 24612801 6 56 gly deglycosylated 795:808 arg1 CN 810:811 deglycosylated CN 795:811 deglycosylated CN obtained by o-glycosidase treatment 795:847 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 8 57 theme heated 1139:1144 arg1 κ-CN 1146:1149 the heated κ-CN 1135:1149 the heated κ-CN 1135:1149 Although the heated κ-CN retained inhibitory activity in a neutralization assay, the activity was weaker than that observed before heat treatment. 24612801 9 58 theme glycan 1399:1404 arg1 residues 1406:1413 glycan residues 1399:1413 glycan residues 1399:1413 Our findings indicate that the inhibitory mechanism of bovine κ-CN against HRV involves direct binding to viral particles via glycan residues. 24612801 3 59 theme late 444:447 arg1 colostrum 449:457 late colostrum 444:457 late colostrum (6-7 d after parturition) 444:483 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 3 60 theme molecular 374:382 arg1 basis 384:388 the molecular basis 370:388 the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk 370:504 The present study examined the molecular basis for the protective effect of bovine κ-CN derived from late colostrum (6-7 d after parturition) and from mature milk. 24612801 4 61 theme casein 531:536 arg1 components 517:526 the components 513:526 the components of casein 513:536 Among the components of casein, κ-CN is the only glycosylated protein that has been identified. 24612801 8 62 theme inhibitory 1160:1169 arg1 activity 1171:1178 inhibitory activity 1160:1178 inhibitory activity 1160:1178 Although the heated κ-CN retained inhibitory activity in a neutralization assay, the activity was weaker than that observed before heat treatment. 24612801 6 63 theme antiviral 856:864 arg1 activity 866:873 antiviral activity 856:873 antiviral activity 856:873 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 24612801 7 64 theme titer-dependent 1102:1116 arg1 manner 1118:1123 a viral titer-dependent manner 1094:1123 a viral titer-dependent manner 1094:1123 Furthermore, an evanescent-field fluorescence-assisted assay showed that HRV particles directly bound to heated casein (at 95°C for 30 min) in a viral titer-dependent manner. 24612801 6 65 theme CN 947:948 arg1 activity 935:942 the antiviral activity 921:942 the antiviral activity of CN 921:948 Desialylated CN obtained by neuraminidase treatment exhibited anti-HRV activity, whereas deglycosylated CN obtained by o-glycosidase treatment lacked antiviral activity, indicating that glycans were responsible for the antiviral activity of CN. 26257376 5 0 theme glucosyl 911:918 arg1 donors 920:925 glucosyl donors 911:925 glucosyl donors 911:925 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 2 1 theme motifs 378:383 arg1 introduction 341:352 the introduction 337:352 the introduction of the 2'- and 2"-deoxy motifs the most efficient way 337:406 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 4 2 theme end 818:820 arg1 linkages 841:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 2 3 dep 2'- 361:363 arg1 way 404:406 the most efficient way 385:406 the most efficient way 385:406 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 2 4 theme -deoxy 371:376 arg1 motifs 378:383 2"-deoxy motifs 369:383 2"-deoxy motifs 369:383 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 1 5 theme α-D-Glcp- 266:274 arg1 -α-D-ManpOMe 299:310 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 248:310 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 248:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 2 6 theme trisaccharide 604:616 arg1 level 618:622 the di- or trisaccharide level 593:622 level 618:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 0 7 theme → 93:93 arg1 α-D-Glcp- 81:89 the trisaccharide α-D-Glcp- 63:89 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2) 63:113 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 0 7 theme → 93:93 arg1 -α-D-Manp- 97:106 1 → 3)-α-D-Manp-(1 → 2 91:112 1 → 3)-α-D-Manp-(1 → 2 91:112 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 5 8 theme promoter 1041:1048 arg1 structure 1017:1025 donor/acceptor structure and nature 1002:1036 structure 1017:1025 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 5 8 theme promoter 1041:1048 arg1 nature 1031:1036 donor/acceptor structure and nature 1002:1036 nature 1031:1036 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 3 9 theme " 643:643 arg1 functions 658:666 the 4"- and 6-deoxy functions 638:666 the 4"- and 6-deoxy functions 638:666 In contrast, the 4"- and 6-deoxy functions were introduced already at the monosaccharide stage. 26257376 1 10 dep analogs 237:243 arg1 " 210:210 2" 209:210 2"- 209:211 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 10 dep analogs 237:243 arg1 2'- 214:216 2'- 214:216 2'- 214:216 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 10 dep analogs 237:243 arg1 6- 223:224 6- 223:224 6- 223:224 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 10 dep analogs 237:243 arg1 " 205:205 4" 204:205 4"- 204:206 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 5 11 from effect 879:884 arg1 donors 920:925 glucosyl donors 911:925 glucosyl donors 911:925 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 5 12 theme individual 1094:1103 arg1 glycosylation 1105:1117 each individual glycosylation 1089:1117 each individual glycosylation 1089:1117 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 4 13 theme most 725:728 arg1 part 742:745 The most challenging part 721:745 The most challenging part of the syntheses 721:762 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 4 13 theme most 725:728 arg1 formation 788:796 the stereoselective formation 768:796 the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages 768:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 1 14 theme → 278:278 arg1 α-D-Glcp- 266:274 trisaccharide α-D-Glcp- 252:274 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 248:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 14 theme → 278:278 arg1 -α-D-Manp- 282:291 1 → 3)-α-D-Manp-(1 → 2 276:297 1 → 3)-α-D-Manp-(1 → 2 276:297 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 4 15 theme linkages 841:848 arg1 part 742:745 The most challenging part 721:745 The most challenging part of the syntheses 721:762 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 4 15 theme linkages 841:848 arg1 formation 788:796 the stereoselective formation 768:796 the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages 768:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 5 16 theme 3-O-acetyl 891:900 arg1 group 902:906 a 3-O-acetyl group 889:906 a 3-O-acetyl group 889:906 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 2 17 theme efficient 394:402 arg1 way 404:406 the most efficient way 385:406 the most efficient way 385:406 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 2 18 theme silver 452:457 arg1 couplings 477:485 silver triflate-promoted couplings 452:485 silver triflate-promoted couplings 452:485 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 3 19 theme 6-deoxy 650:656 arg1 functions 658:666 the 4"- and 6-deoxy functions 638:666 the 4"- and 6-deoxy functions 638:666 In contrast, the 4"- and 6-deoxy functions were introduced already at the monosaccharide stage. 26257376 5 20 theme group 902:906 arg1 effect 879:884 The established α-directing effect 851:884 The established α-directing effect of a 3-O-acetyl group in glucosyl donors 851:925 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 2 21 theme 2-deoxy 576:582 arg1 motif 584:588 a 2-deoxy motif 574:588 a 2-deoxy motif at the di- or trisaccharide level 574:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 0 22 dep -α-D-Manp- 97:106 arg1 3 95:95 3 95:95 3 95:95 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 4 23 theme non-reducing 805:816 arg1 linkages 841:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 1 24 theme efficient 175:183 arg1 synthesis 185:193 efficient synthesis 175:193 efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 175:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 0 25 theme monodeoxy 42:50 arg1 analogs 52:58 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs 13:58 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2) 13:113 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 2 26 theme 1,2-di-bromo-mannosyl 421:441 arg1 donor 443:447 a 1,2-di-bromo-mannosyl donor 419:447 a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings 419:485 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 3 27 theme monosaccharide 699:712 arg1 stage 714:718 the monosaccharide stage 695:718 the monosaccharide stage 695:718 In contrast, the 4"- and 6-deoxy functions were introduced already at the monosaccharide stage. 26257376 5 28 theme effect 965:970 arg1 variable 976:983 variable 976:983 variable 976:983 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 5 28 theme effect 965:970 arg1 magnitude 948:956 the magnitude 944:956 the magnitude of the effect 944:970 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 2 29 from level 618:622 arg1 motif 584:588 a 2-deoxy motif 574:588 a 2-deoxy motif at the di- or trisaccharide level 574:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 5 30 theme established 855:865 arg1 effect 879:884 The established α-directing effect 851:884 The established α-directing effect of a 3-O-acetyl group in glucosyl donors 851:925 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 2 31 theme 2-bromo-function 554:569 arg1 reduction 537:545 reduction 537:545 reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level 537:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 4 32 theme syntheses 754:762 arg1 part 742:745 The most challenging part 721:745 The most challenging part of the syntheses 721:762 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 4 32 theme syntheses 754:762 arg1 formation 788:796 the stereoselective formation 768:796 the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages 768:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 0 33 dep -α-D-ManpOMe 114:125 arg1 recognized 127:136 recognized 127:136 -α-D-ManpOMe recognized by Calreticulin/Calnexin 114:161 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 5 34 theme α-directing 867:877 arg1 effect 879:884 The established α-directing effect 851:884 The established α-directing effect of a 3-O-acetyl group in glucosyl donors 851:925 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 4 35 theme stereoselective 772:786 arg1 part 742:745 The most challenging part 721:745 The most challenging part of the syntheses 721:762 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 4 35 theme stereoselective 772:786 arg1 formation 788:796 the stereoselective formation 768:796 the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages 768:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 0 36 theme trisaccharide 67:79 arg1 α-D-Glcp- 81:89 the trisaccharide α-D-Glcp- 63:89 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2) 63:113 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 0 36 theme trisaccharide 67:79 arg1 -α-D-Manp- 97:106 1 → 3)-α-D-Manp-(1 → 2 91:112 1 → 3)-α-D-Manp-(1 → 2 91:112 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 1 37 theme monodeoxy 227:235 arg1 analogs 237:243 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs 198:243 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 198:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 0 38 theme analogs 52:58 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2) 0:113 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 2 39 theme 2'- 361:363 arg1 introduction 341:352 the introduction 337:352 the introduction of the 2'- and 2"-deoxy motifs the most efficient way 337:406 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 1 40 theme analogs 237:243 arg1 synthesis 185:193 efficient synthesis 175:193 efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 175:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 2 41 theme di- 597:599 arg1 level 618:622 the di- or trisaccharide level 593:622 level 618:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 2 42 theme triflate-promoted 459:475 arg1 couplings 477:485 silver triflate-promoted couplings 452:485 silver triflate-promoted couplings 452:485 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 5 43 theme donor/acceptor 1002:1015 arg1 structure 1017:1025 donor/acceptor structure and nature 1002:1036 structure 1017:1025 The established α-directing effect of a 3-O-acetyl group in glucosyl donors was explored but the magnitude of the effect was variable and dependent on donor/acceptor structure and nature of promoter and an optimization had to be made for each individual glycosylation. 26257376 0 44 theme α-D-Glcp- 81:89 arg1 analogs 52:58 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs 13:58 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2) 13:113 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 1 45 theme -α-D-ManpOMe 299:310 arg1 analogs 237:243 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs 198:243 four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 198:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 46 theme trisaccharide 252:264 arg1 α-D-Glcp- 266:274 trisaccharide α-D-Glcp- 252:274 the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe 248:310 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 1 46 theme trisaccharide 252:264 arg1 -α-D-Manp- 282:291 1 → 3)-α-D-Manp-(1 → 2 276:297 1 → 3)-α-D-Manp-(1 → 2 276:297 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26257376 4 47 theme cis-α-D-glucosidic 822:839 arg1 linkages 841:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 the non-reducing end cis-α-D-glucosidic linkages 801:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 2 48 from donor 443:447 arg1 couplings 477:485 silver triflate-promoted couplings 452:485 silver triflate-promoted couplings 452:485 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 4 49 theme challenging 730:740 arg1 part 742:745 The most challenging part 721:745 The most challenging part of the syntheses 721:762 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 4 49 theme challenging 730:740 arg1 formation 788:796 the stereoselective formation 768:796 the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages 768:848 The most challenging part of the syntheses was the stereoselective formation of the non-reducing end cis-α-D-glucosidic linkages. 26257376 2 50 theme α-glycosidic 504:515 arg1 linkage 517:523 the α-glycosidic linkage 500:523 the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level 500:622 For the introduction of the 2'- and 2"-deoxy motifs the most efficient way was to use a 1,2-di-bromo-mannosyl donor in silver triflate-promoted couplings to construct the α-glycosidic linkage followed by reduction of the 2-bromo-function to a 2-deoxy motif at the di- or trisaccharide level. 26257376 0 51 dep analogs 52:58 arg1 " 20:20 4" 19:20 4"- 19:21 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 0 51 dep analogs 52:58 arg1 " 25:25 2" 24:25 2"- 24:26 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 0 51 dep analogs 52:58 arg1 2'- 29:31 2'- 29:31 2'- 29:31 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 0 51 dep analogs 52:58 arg1 6- 38:39 6- 38:39 6- 38:39 Synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe recognized by Calreticulin/Calnexin. 26257376 1 52 dep -α-D-Manp- 282:291 arg1 3 280:280 3 280:280 3 280:280 Routes for efficient synthesis of four (4"-, 2"-, 2'-, and 6-) monodeoxy analogs of the trisaccharide α-D-Glcp-(1 → 3)-α-D-Manp-(1 → 2)-α-D-ManpOMe have been developed. 26001781 0 0 theme Residue 71:77 arg1 Addition 8:15 Limited Addition 0:15 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 0:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 2 1 theme Arabidopsis 310:320 arg1 thaliana 322:329 Arabidopsis thaliana 310:329 Arabidopsis thaliana 310:329 Here, we report a mechanism in Arabidopsis thaliana that efficiently produces the largest N-glycan in plants. 26001781 5 2 theme GlcNAc 1129:1134 arg1 GlcNAc 1148:1153 (GlcNAc)2Man3XylFuc(GlcNAc)2 1128:1155 (GlcNAc)2Man3XylFuc(GlcNAc)2 1128:1155 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 3 theme core 865:868 arg1 β1,2-xylose 870:880 the core β1,2-xylose and α1,3-fucose residues 861:905 β1,2-xylose 870:880 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 5 4 theme prevalent 1079:1087 arg1 N-glycans 1089:1097 the prevalent N-glycans 1075:1097 the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1075:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 5 5 theme addition 964:971 arg1 rate 952:955 the rate 948:955 the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor 948:1031 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 5 5 theme addition 964:971 arg1 mechanism 1038:1046 a mechanism 1036:1046 a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1036:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 6 theme transgenic 704:713 arg1 plants 715:720 35S:GnTII transgenic plants 694:720 35S:GnTII transgenic plants 694:720 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 1 7 theme Man3XylFuc 257:266 arg1 GlcNAc 268:273 Man3XylFuc(GlcNAc)2 257:275 Man3XylFuc(GlcNAc)2 257:275 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 7 theme Man3XylFuc 257:266 arg1 residues 247:254 α1,3-fucose residues 235:254 α1,3-fucose residues (Man3XylFuc(GlcNAc)2) 235:276 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 4 8 theme residues 898:905 arg1 additions 848:856 additions 848:856 additions of the core β1,2-xylose and α1,3-fucose residues 848:905 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 5 9 with N-glycans 1089:1097 arg1 structures 1157:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 5 10 theme 2Man3XylFuc 1136:1146 arg1 GlcNAc 1148:1153 (GlcNAc)2Man3XylFuc(GlcNAc)2 1128:1155 (GlcNAc)2Man3XylFuc(GlcNAc)2 1128:1155 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 11 theme mutant 683:688 arg1 analysis 666:673 analysis 666:673 analysis of gnt2 mutant and 35S:GnTII transgenic plants 666:720 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 4 12 theme N-glycan 801:808 arg1 GlcNAc 830:835 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 2 13 from mechanism 297:305 arg1 thaliana 322:329 Arabidopsis thaliana 310:329 Arabidopsis thaliana 310:329 Here, we report a mechanism in Arabidopsis thaliana that efficiently produces the largest N-glycan in plants. 26001781 4 14 theme common 794:799 arg1 GlcNAc 830:835 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 2 15 theme largest 361:367 arg1 N-glycan 369:376 the largest N-glycan 357:376 the largest N-glycan in plants 357:386 Here, we report a mechanism in Arabidopsis thaliana that efficiently produces the largest N-glycan in plants. 26001781 4 16 theme GlcNAcMan3 819:828 arg1 GlcNAc 830:835 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 4 17 theme GnTII 698:702 arg1 plants 715:720 35S:GnTII transgenic plants 694:720 35S:GnTII transgenic plants 694:720 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 0 18 theme Limited 0:6 arg1 Addition 8:15 Limited Addition 0:15 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 0:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 5 19 theme GlcNAc 1115:1120 arg1 structures 1157:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 20 theme acceptor 810:817 arg1 GlcNAc 830:835 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 790:837 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 0 21 link β1,2-linked 30:40 arg1 Residue 71:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 3 22 theme biochemical 401:411 arg1 evidence 413:420 Genetic and biochemical evidence 389:420 Genetic and biochemical evidence 389:420 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 4 23 theme non-reducing 759:770 arg1 residue 779:785 the 6-arm non-reducing GlcNAc residue 749:785 the 6-arm non-reducing GlcNAc residue 749:785 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 1 24 with N-glycan 200:207 arg1 GlcNAc 268:273 Man3XylFuc(GlcNAc)2 257:275 Man3XylFuc(GlcNAc)2 257:275 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 24 with N-glycan 200:207 arg1 β1,2-xylose 219:229 core β1,2-xylose 214:229 core β1,2-xylose 214:229 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 24 with N-glycan 200:207 arg1 residues 247:254 α1,3-fucose residues 235:254 α1,3-fucose residues (Man3XylFuc(GlcNAc)2) 235:276 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 5 25 theme 6-arm 980:984 arg1 residue 993:999 the 6-arm GlcNAc residue 976:999 the 6-arm GlcNAc residue 976:999 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 26 theme plants 715:720 arg1 analysis 666:673 analysis 666:673 analysis of gnt2 mutant and 35S:GnTII transgenic plants 666:720 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 4 27 theme residue 779:785 arg1 addition 737:744 the addition 733:744 the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 733:837 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 0 28 theme Largest 112:118 arg1 N-Glycan 120:127 the Largest N-Glycan 108:127 the Largest N-Glycan in Plants 108:137 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 5 29 theme common 1008:1013 arg1 acceptor 1024:1031 the common N-glycan acceptor 1004:1031 the common N-glycan acceptor 1004:1031 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 1 30 theme abundant 149:156 arg1 N-glycan 158:165 The most abundant N-glycan 140:165 The most abundant N-glycan in plants 140:175 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 30 theme abundant 149:156 arg1 N-glycan 200:207 the paucimannosidic N-glycan 180:207 the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2) 180:276 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 31 theme paucimannosidic 184:198 arg1 N-glycan 158:165 The most abundant N-glycan 140:165 The most abundant N-glycan in plants 140:175 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 31 theme paucimannosidic 184:198 arg1 N-glycan 200:207 the paucimannosidic N-glycan 180:207 the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2) 180:276 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 0 32 theme β1,2-linked 30:40 arg1 Residue 71:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 4 33 theme gnt2 678:681 arg1 mutant 683:688 gnt2 mutant 678:688 gnt2 mutant 678:688 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 4 34 theme α1,3-fucose 886:896 arg1 residues 898:905 the core β1,2-xylose and α1,3-fucose residues 861:905 residues 898:905 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 5 35 theme N-glycan 1015:1022 arg1 acceptor 1024:1031 the common N-glycan acceptor 1004:1031 the common N-glycan acceptor 1004:1031 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 3 36 theme β1,2-xylose 574:584 arg1 additions 552:560 additions 552:560 additions of the core β1,2-xylose and α1,3-fucose residues 552:609 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 0 37 theme 6-Arm 24:28 arg1 Residue 71:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 5 38 theme N-glycans 1089:1097 arg1 formation 1062:1070 formation 1062:1070 formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1062:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 3 39 theme Genetic 389:395 arg1 evidence 413:420 Genetic and biochemical evidence 389:420 Genetic and biochemical evidence 389:420 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 4 40 theme GlcNAc 772:777 arg1 residue 779:785 the 6-arm non-reducing GlcNAc residue 749:785 the 6-arm non-reducing GlcNAc residue 749:785 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 3 41 theme β1,2-GlcNAc 463:473 arg1 residue 475:481 the 6-arm β1,2-GlcNAc residue 453:481 the 6-arm β1,2-GlcNAc residue 453:481 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 0 42 theme N-Acetylglucosamine 42:60 arg1 Residue 71:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 5 43 theme Man3XylFuc 1104:1113 arg1 GlcNAc 1115:1120 Man3XylFuc(GlcNAc)2 1104:1122 Man3XylFuc(GlcNAc)2 1104:1122 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 0 44 theme N-Glycan 120:127 arg1 Formation 95:103 the Formation 91:103 the Formation of the Largest N-Glycan in Plants 91:137 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 5 45 theme GlcNAc 986:991 arg1 residue 993:999 the 6-arm GlcNAc residue 976:999 the 6-arm GlcNAc residue 976:999 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 3 46 theme core 569:572 arg1 β1,2-xylose 574:584 the core β1,2-xylose and α1,3-fucose residues 565:609 β1,2-xylose 574:584 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 2 47 from N-glycan 369:376 arg1 plants 381:386 plants 381:386 plants 381:386 Here, we report a mechanism in Arabidopsis thaliana that efficiently produces the largest N-glycan in plants. 26001781 3 48 theme residue 475:481 arg1 addition 441:448 the addition 437:448 the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) 437:527 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 3 48 theme residue 475:481 arg1 effective 537:545 effective 537:545 effective 537:545 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 4 49 theme 6-arm 753:757 arg1 residue 779:785 the 6-arm non-reducing GlcNAc residue 749:785 the 6-arm non-reducing GlcNAc residue 749:785 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 5 50 theme residue 993:999 arg1 addition 964:971 the addition 960:971 the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor 960:1031 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 51 theme 35S 694:696 arg1 plants 715:720 35S:GnTII transgenic plants 694:720 35S:GnTII transgenic plants 694:720 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 0 52 from Plants 132:137 arg1 Formation 95:103 the Formation 91:103 the Formation of the Largest N-Glycan in Plants 91:137 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 0 53 from Formation 95:103 arg1 Plants 132:137 Plants 132:137 Plants 132:137 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 3 54 theme 6-arm 457:461 arg1 residue 475:481 the 6-arm β1,2-GlcNAc residue 453:481 the 6-arm β1,2-GlcNAc residue 453:481 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 5 55 theme GlcNAc 1148:1153 arg1 structures 1157:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures 1104:1166 Our findings indicate that plants limit the rate of the addition of the 6-arm GlcNAc residue to the common N-glycan acceptor as a mechanism to facilitate formation of the prevalent N-glycans with Man3XylFuc(GlcNAc)2 and (GlcNAc)2Man3XylFuc(GlcNAc)2 structures. 26001781 4 56 theme β1,2-xylose 870:880 arg1 additions 848:856 additions 848:856 additions of the core β1,2-xylose and α1,3-fucose residues 848:905 Furthermore, analysis of gnt2 mutant and 35S:GnTII transgenic plants shows that the addition of the 6-arm non-reducing GlcNAc residue to the common N-glycan acceptor GlcNAcMan3(GlcNAc)2 inhibits additions of the core β1,2-xylose and α1,3-fucose residues. 26001781 3 57 theme α1,3-fucose 590:600 arg1 residues 602:609 the core β1,2-xylose and α1,3-fucose residues 565:609 residues 602:609 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 0 58 from N-Glycan 120:127 arg1 Plants 132:137 Plants 132:137 Plants 132:137 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 0 59 theme GlcNAc 63:68 arg1 Residue 71:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue 20:77 Limited Addition of the 6-Arm β1,2-linked N-Acetylglucosamine (GlcNAc) Residue Facilitates the Formation of the Largest N-Glycan in Plants. 26001781 3 60 theme residues 602:609 arg1 additions 552:560 additions 552:560 additions of the core β1,2-xylose and α1,3-fucose residues 552:609 Genetic and biochemical evidence indicates that the addition of the 6-arm β1,2-GlcNAc residue by N-acetylglucosaminyltransferase II (GnTII) is less effective than additions of the core β1,2-xylose and α1,3-fucose residues by XylT, FucTA, and FucTB in Arabidopsis. 26001781 1 61 from N-glycan 158:165 arg1 plants 170:175 plants 170:175 plants 170:175 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 62 theme core 214:217 arg1 β1,2-xylose 219:229 core β1,2-xylose 214:229 core β1,2-xylose 214:229 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 63 theme α1,3-fucose 235:245 arg1 GlcNAc 268:273 Man3XylFuc(GlcNAc)2 257:275 Man3XylFuc(GlcNAc)2 257:275 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 26001781 1 63 theme α1,3-fucose 235:245 arg1 residues 247:254 α1,3-fucose residues 235:254 α1,3-fucose residues (Man3XylFuc(GlcNAc)2) 235:276 The most abundant N-glycan in plants is the paucimannosidic N-glycan with core β1,2-xylose and α1,3-fucose residues (Man3XylFuc(GlcNAc)2). 24359113 9 0 theme galactosemia 1401:1412 arg1 patients 1414:1421 galactosemia patients 1401:1421 galactosemia patients 1401:1421 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 7 1 theme N-glycan 1178:1185 arg1 defects 1198:1204 N-glycan processing defects 1178:1204 N-glycan processing defects in children with galactosemia 1178:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 6 2 theme processing 1055:1064 arg1 defects 1066:1072 ongoing N-glycan processing defects 1038:1072 ongoing N-glycan processing defects 1038:1072 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 0 3 from abnormalities 9:21 arg1 children 26:33 children 26:33 children with galactosemia 26:51 N-glycan abnormalities in children with galactosemia. 24359113 7 4 theme processing 1187:1196 arg1 defects 1198:1204 N-glycan processing defects 1178:1204 N-glycan processing defects in children with galactosemia 1178:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 5 5 theme dietary 939:945 arg1 restriction 947:957 dietary restriction 939:957 dietary restriction of galactose 939:970 We have observed that severe N-glycan assembly defects correct in the neonate following dietary restriction of galactose. 24359113 1 6 from intoxication 64:75 arg1 galactosemia 101:112 galactosemia 101:112 galactosemia 101:112 Galactose intoxication and over-restriction in galactosemia may affect glycosylation pathways and cause multisystem effects. 24359113 6 7 theme N-glycan 1046:1053 arg1 defects 1066:1072 ongoing N-glycan processing defects 1038:1072 ongoing N-glycan processing defects 1038:1072 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 3 8 with child 618:622 arg1 variant 638:644 a milder variant 629:644 a milder variant (genotype S135L/S135L) 629:667 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 8 with child 618:622 arg1 S135L/S135L 656:666 genotype S135L/S135L 647:666 genotype S135L/S135L 647:666 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 4 9 theme pediatric 824:832 arg1 samples 842:848 nine pediatric control samples 819:848 nine pediatric control samples 819:848 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 3 10 with children 553:560 arg1 child 618:622 one child 614:622 one child with a milder variant (genotype S135L/S135L) 614:667 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 10 with children 553:560 arg1 galactosemia 574:585 severe galactosemia 567:585 severe galactosemia (genotype Q188R/Q188R) 567:608 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 10 with children 553:560 arg1 Q188R/Q188R 597:607 genotype Q188R/Q188R 588:607 genotype Q188R/Q188R 588:607 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 2 11 theme extracted 346:354 arg1 N-glycans 360:368 extracted IgG N-glycans 346:368 extracted IgG N-glycans 346:368 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 1 12 theme glycosylation 125:137 arg1 pathways 139:146 glycosylation pathways 125:146 glycosylation pathways 125:146 Galactose intoxication and over-restriction in galactosemia may affect glycosylation pathways and cause multisystem effects. 24359113 6 13 theme ongoing 1038:1044 arg1 defects 1066:1072 ongoing N-glycan processing defects 1038:1072 ongoing N-glycan processing defects 1038:1072 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 4 14 theme congenital 769:778 arg1 disorder 780:787 congenital disorder 769:787 congenital disorder of glycosylation type Ia 769:812 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 4 15 theme glycosylation 792:804 arg1 type 806:809 glycosylation type Ia 792:812 glycosylation type Ia 792:812 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 3 16 theme genotype 588:595 arg1 galactosemia 574:585 severe galactosemia 567:585 severe galactosemia (genotype Q188R/Q188R) 567:608 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 16 theme genotype 588:595 arg1 Q188R/Q188R 597:607 genotype Q188R/Q188R 588:607 genotype Q188R/Q188R 588:607 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 8 17 theme processing 1260:1269 arg1 defects 1271:1277 N-glycan processing defects 1251:1277 N-glycan processing defects present in galactosemia children from an early age 1251:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 3 18 theme milder 631:636 arg1 variant 638:644 a milder variant 629:644 a milder variant (genotype S135L/S135L) 629:667 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 18 theme milder 631:636 arg1 S135L/S135L 656:666 genotype S135L/S135L 647:666 genotype S135L/S135L 647:666 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 8 19 from children 1303:1310 arg1 present 1279:1285 present 1279:1285 present 1279:1285 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 3 20 theme severe 567:572 arg1 galactosemia 574:585 severe galactosemia 567:585 severe galactosemia (genotype Q188R/Q188R) 567:608 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 20 theme severe 567:572 arg1 Q188R/Q188R 597:607 genotype Q188R/Q188R 588:607 genotype Q188R/Q188R 588:607 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 5 21 theme assembly 889:896 arg1 defects 898:904 severe N-glycan assembly defects 873:904 severe N-glycan assembly defects 873:904 We have observed that severe N-glycan assembly defects correct in the neonate following dietary restriction of galactose. 24359113 7 22 from presence 1166:1173 arg1 children 1209:1216 children 1209:1216 children with galactosemia 1209:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 8 23 theme galactosemia 1290:1301 arg1 children 1303:1310 galactosemia children 1290:1310 galactosemia children from an early age 1290:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 4 24 with children 745:752 arg1 samples 842:848 nine pediatric control samples 819:848 nine pediatric control samples 819:848 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 4 24 with children 745:752 arg1 PMM2-CDG 759:766 PMM2-CDG 759:766 PMM2-CDG (congenital disorder of glycosylation type Ia) 759:813 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 2 25 theme whole 330:334 arg1 serum 336:340 whole serum 330:340 whole serum 330:340 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 8 26 from present 1279:1285 arg1 children 1303:1310 galactosemia children 1290:1310 galactosemia children from an early age 1290:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 2 27 theme quantitative 479:490 arg1 measure 492:498 a quantitative measure 477:498 a quantitative measure of galactose incorporation 477:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 8 28 theme N-glycan 1251:1258 arg1 defects 1271:1277 N-glycan processing defects 1251:1277 N-glycan processing defects present in galactosemia children from an early age 1251:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 2 29 dep agalactosylated 390:404 arg1 G0 407:408 G0 407:408 G0 407:408 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 9 30 theme /G2 1360:1362 arg1 ratios 1364:1369 (G0/G1)/G2 ratios 1353:1369 (G0/G1)/G2 ratios 1353:1369 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 0 31 theme N-glycan 0:7 arg1 abnormalities 9:21 N-glycan abnormalities 0:21 N-glycan abnormalities in children with galactosemia. 0:52 N-glycan abnormalities in children with galactosemia. 24359113 8 32 theme early 1320:1324 arg1 age 1326:1328 an early age 1317:1328 an early age 1317:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 8 33 theme present 1279:1285 arg1 defects 1271:1277 N-glycan processing defects 1251:1277 N-glycan processing defects present in galactosemia children from an early age 1251:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 8 34 attach present 1279:1285 arg1 children 1303:1310 galactosemia children 1290:1310 galactosemia children from an early age 1290:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 8 34 attach present 1279:1285 arg2 defects 1271:1277 N-glycan processing defects 1251:1277 N-glycan processing defects present in galactosemia children from an early age 1251:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 2 35 theme structures 463:472 arg1 measurement 375:385 measurement 375:385 measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 375:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 7 36 with children 1209:1216 arg1 galactosemia 1223:1234 galactosemia 1223:1234 galactosemia 1223:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 2 37 theme IgG 356:358 arg1 N-glycans 360:368 extracted IgG N-glycans 346:368 extracted IgG N-glycans 346:368 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 2 38 theme incorporation 513:525 arg1 measure 492:498 a quantitative measure 477:498 a quantitative measure of galactose incorporation 477:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 6 39 theme galactosemia 996:1007 arg1 patients 1009:1016 treated adult galactosemia patients 982:1016 treated adult galactosemia patients 982:1016 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 7 40 theme incorporation 1117:1129 arg1 ratios 1131:1136 informative galactose incorporation ratios 1095:1136 informative galactose incorporation ratios 1095:1136 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 7 40 theme incorporation 1117:1129 arg1 method 1143:1148 a method 1141:1148 a method of studying the presence of N-glycan processing defects in children with galactosemia 1141:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 2 41 theme chromatography 281:294 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 5 42 theme severe 873:878 arg1 defects 898:904 severe N-glycan assembly defects 873:904 severe N-glycan assembly defects 873:904 We have observed that severe N-glycan assembly defects correct in the neonate following dietary restriction of galactose. 24359113 2 43 theme galactose 503:511 arg1 incorporation 513:525 galactose incorporation 503:525 galactose incorporation 503:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 6 44 theme adult 990:994 arg1 patients 1009:1016 treated adult galactosemia patients 982:1016 treated adult galactosemia patients 982:1016 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 3 45 with galactosemia 574:585 arg1 variant 638:644 a milder variant 629:644 a milder variant (genotype S135L/S135L) 629:667 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 45 with galactosemia 574:585 arg1 S135L/S135L 656:666 genotype S135L/S135L 647:666 genotype S135L/S135L 647:666 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 2 46 theme liquid 274:279 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 0 47 with children 26:33 arg1 galactosemia 40:51 galactosemia 40:51 galactosemia 40:51 N-glycan abnormalities in children with galactosemia. 24359113 2 48 theme digalactosylated 441:456 arg1 structures 463:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures 390:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 390:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 7 49 theme defects 1198:1204 arg1 presence 1166:1173 the presence 1162:1173 the presence of N-glycan processing defects in children with galactosemia 1162:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 6 50 theme treated 982:988 arg1 patients 1009:1016 treated adult galactosemia patients 982:1016 treated adult galactosemia patients 982:1016 However, treated adult galactosemia patients continue to exhibit ongoing N-glycan processing defects. 24359113 7 51 theme informative 1095:1105 arg1 ratios 1131:1136 informative galactose incorporation ratios 1095:1136 informative galactose incorporation ratios 1095:1136 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 7 51 theme informative 1095:1105 arg1 method 1143:1148 a method 1141:1148 a method of studying the presence of N-glycan processing defects in children with galactosemia 1141:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 2 52 theme ultra-performance 256:272 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 8 53 from age 1326:1328 arg1 children 1303:1310 galactosemia children 1290:1310 galactosemia children from an early age 1290:1328 We identified N-glycan processing defects present in galactosemia children from an early age. 24359113 9 54 theme G0/G1 1354:1358 arg1 ratios 1364:1369 (G0/G1)/G2 ratios 1353:1369 (G0/G1)/G2 ratios 1353:1369 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 7 55 theme galactose 1107:1115 arg1 ratios 1131:1136 informative galactose incorporation ratios 1095:1136 informative galactose incorporation ratios 1095:1136 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 7 55 theme galactose 1107:1115 arg1 method 1143:1148 a method 1141:1148 a method of studying the presence of N-glycan processing defects in children with galactosemia 1141:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 2 56 theme interaction 229:239 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 2 57 theme chromatography 241:254 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 4 58 theme age-matched 733:743 arg1 children 745:752 three age-matched children 727:752 three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples 727:848 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 1 59 theme Galactose 54:62 arg1 intoxication 64:75 Galactose intoxication 54:75 Galactose intoxication 54:75 Galactose intoxication and over-restriction in galactosemia may affect glycosylation pathways and cause multisystem effects. 24359113 2 60 theme hydrophilic 217:227 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 3 61 theme genotype 647:654 arg1 variant 638:644 a milder variant 629:644 a milder variant (genotype S135L/S135L) 629:667 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 3 61 theme genotype 647:654 arg1 S135L/S135L 656:666 genotype S135L/S135L 647:666 genotype S135L/S135L 647:666 This was applied to nine children with severe galactosemia (genotype Q188R/Q188R) and one child with a milder variant (genotype S135L/S135L). 24359113 2 62 dep digalactosylated 441:456 arg1 G2 459:460 G2 459:460 G2 459:460 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 2 63 theme agalactosylated 390:404 arg1 structures 463:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures 390:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 390:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 4 64 dep PMM2-CDG 759:766 arg1 disorder 780:787 congenital disorder 769:787 congenital disorder of glycosylation type Ia 769:812 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 2 65 dep monogalactosylated 412:429 arg1 G1 432:433 G1 432:433 G1 432:433 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 2 66 theme applied 209:215 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 4 67 theme type 806:809 arg1 disorder 780:787 congenital disorder 769:787 congenital disorder of glycosylation type Ia 769:812 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 1 68 theme multisystem 158:168 arg1 effects 170:176 multisystem effects 158:176 multisystem effects 158:176 Galactose intoxication and over-restriction in galactosemia may affect glycosylation pathways and cause multisystem effects. 24359113 9 69 dep significant 1466:1476 arg1 significant 1466:1476 significant 1466:1476 significant 1466:1476 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 9 69 dep significant 1466:1476 arg1 = 1481:1481 = 1481:1481 = 1481:1481 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 9 69 dep significant 1466:1476 arg1 difference 1376:1385 the difference 1372:1385 the difference noted between galactosemia patients and controls 1372:1434 For G0/G1, G0/G2, and (G0/G1)/G2 ratios, the difference noted between galactosemia patients and controls was found to be statistically significant (p = 0.002, 0.01, and 0.006, respectively). 24359113 5 70 theme galactose 962:970 arg1 restriction 947:957 dietary restriction 939:957 dietary restriction of galactose 939:970 We have observed that severe N-glycan assembly defects correct in the neonate following dietary restriction of galactose. 24359113 5 71 theme N-glycan 880:887 arg1 defects 898:904 severe N-glycan assembly defects 873:904 severe N-glycan assembly defects 873:904 We have observed that severe N-glycan assembly defects correct in the neonate following dietary restriction of galactose. 24359113 4 72 theme control 834:840 arg1 samples 842:848 nine pediatric control samples 819:848 nine pediatric control samples 819:848 The profiles were also compared with those obtained from three age-matched children with PMM2-CDG (congenital disorder of glycosylation type Ia) and nine pediatric control samples. 24359113 7 73 from children 1209:1216 arg1 presence 1166:1173 the presence 1162:1173 the presence of N-glycan processing defects in children with galactosemia 1162:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 7 74 from defects 1198:1204 arg1 children 1209:1216 children 1209:1216 children with galactosemia 1209:1234 We have now applied informative galactose incorporation ratios as a method of studying the presence of N-glycan processing defects in children with galactosemia. 24359113 2 75 theme high-throughput 296:310 arg1 method 312:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method 206:317 an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 206:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 2 76 theme monogalactosylated 412:429 arg1 structures 463:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures 390:472 agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation 390:525 In this study, we describe an applied hydrophilic interaction chromatography ultra-performance liquid chromatography high-throughput method to analyze whole serum and extracted IgG N-glycans with measurement of agalactosylated (G0), monogalactosylated (G1), and digalactosylated (G2) structures as a quantitative measure of galactose incorporation. 24359113 1 77 from over-restriction 81:96 arg1 galactosemia 101:112 galactosemia 101:112 galactosemia 101:112 Galactose intoxication and over-restriction in galactosemia may affect glycosylation pathways and cause multisystem effects. 24854630 3 0 theme reproducible 545:556 arg1 changes 558:564 reproducible changes 545:564 reproducible changes in expression of glycoprotein synthetic enzymes 545:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 1 1 theme several 250:256 arg1 malignancies 258:269 several malignancies 250:269 several malignancies including prostate cancer 250:295 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 1 theme several 250:256 arg1 cancer 290:295 prostate cancer 281:295 prostate cancer 281:295 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 8 2 theme MUC1 1154:1157 arg1 antibodies 1167:1176 PSA, PAP, and MUC1 capture antibodies 1140:1176 antibodies 1167:1176 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 3 3 from changes 558:564 arg1 expression 569:578 expression 569:578 expression of glycoprotein synthetic enzymes 569:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 3 4 theme synthetic 596:604 arg1 enzymes 606:612 glycoprotein synthetic enzymes 583:612 glycoprotein synthetic enzymes 583:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 7 5 theme tissue 1112:1117 arg1 samples 1119:1125 fresh-frozen prostate tissue samples 1090:1125 fresh-frozen prostate tissue samples 1090:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 2 6 from differences 361:371 arg1 levels 392:397 gene expression levels 376:397 gene expression levels of glycoprotein synthetic enzymes 376:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 7 7 theme fresh-frozen 1090:1101 arg1 samples 1119:1125 fresh-frozen prostate tissue samples 1090:1125 fresh-frozen prostate tissue samples 1090:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 2 8 theme altered 319:325 arg1 glycosylation 327:339 altered glycosylation 319:339 altered glycosylation 319:339 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 1 9 theme posttranslational 167:183 arg1 modification 185:196 a common posttranslational modification 158:196 a common posttranslational modification 158:196 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 9 theme posttranslational 167:183 arg1 glycosylation 141:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 8 10 theme core 1193:1196 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 6 11 theme cancer 956:961 arg1 datasets 979:986 several prostate cancer gene expression datasets 939:986 several prostate cancer gene expression datasets 939:986 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 8 12 from differences 1377:1387 arg1 P = 0.06 1411:1418 P = 0.06 1411:1418 P = 0.06 1411:1418 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 8 12 from differences 1377:1387 arg1 levels 1403:1408 PAP sLe(x) levels 1392:1408 PAP sLe(x) levels (P = 0.06) 1392:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 5 13 theme prostatic 817:825 arg1 PAP 847:849 PAP 847:849 PAP 847:849 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 13 theme prostatic 817:825 arg1 phosphatase 834:844 prostatic acidic phosphatase 817:844 prostatic acidic phosphatase (PAP) 817:850 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 8 14 theme O-linked 1200:1207 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 6 15 theme several 939:945 arg1 datasets 979:986 several prostate cancer gene expression datasets 939:986 several prostate cancer gene expression datasets 939:986 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 8 16 theme sLe 1396:1398 arg1 P = 0.06 1411:1418 P = 0.06 1411:1418 P = 0.06 1411:1418 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 8 16 theme sLe 1396:1398 arg1 levels 1403:1408 PAP sLe(x) levels 1392:1408 PAP sLe(x) levels (P = 0.06) 1392:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 7 17 theme normal 1064:1069 arg1 tissue 1080:1085 normal prostate tissue 1064:1085 normal prostate tissue in fresh-frozen prostate tissue samples 1064:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 7 18 theme transcript 1016:1025 arg1 levels 1027:1032 increased transcript levels 1006:1032 increased transcript levels in cancer samples 1006:1050 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 5 19 theme phosphatase 834:844 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 0 20 from O-glycosylation 57:71 arg1 cancers 113:119 human prostate cancers 98:119 human prostate cancers 98:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 8 21 theme structure 1273:1281 arg1 elevation 1248:1256 elevation 1248:1256 elevation of this glycan structure in cancer 1248:1291 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 11 22 theme glycosylation 1714:1726 arg1 Alterations 1690:1700 Alterations 1690:1700 Alterations of O-linked glycosylation 1690:1726 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 5 23 theme glycan 730:735 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 11 24 theme biomarkers 1862:1871 arg1 development 1809:1819 development 1809:1819 development of prostate cancer specific glycoprotein biomarkers 1809:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 7 25 theme cancer 1037:1042 arg1 samples 1044:1050 cancer samples 1037:1050 cancer samples 1037:1050 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 5 26 theme specific 774:781 arg1 PSA 792:794 PSA 792:794 PSA 792:794 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 26 theme specific 774:781 arg1 antigen 783:789 prostate specific antigen 765:789 prostate specific antigen (PSA) 765:795 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 8 27 from elevation 1248:1256 arg1 cancer 1286:1291 cancer 1286:1291 cancer 1286:1291 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 1 28 theme BACKGROUND 122:131 arg1 modification 185:196 a common posttranslational modification 158:196 a common posttranslational modification 158:196 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 28 theme BACKGROUND 122:131 arg1 glycosylation 141:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 5 29 dep X 750:750 arg1 x 757:757 x 757:757 x 757:757 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 29 dep X 750:750 arg1 sLe 753:755 sLe 753:755 sLe(x) 753:758 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 30 theme Lewis 744:748 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 0 31 theme PSA 76:78 arg1 O-glycosylation 57:71 altered O-glycosylation 49:71 altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers 49:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 11 32 from important 1737:1745 arg1 biology 1766:1772 prostate cancer biology 1750:1772 prostate cancer biology 1750:1772 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 10 33 from proteins 1680:1687 arg1 levels 1633:1638 higher levels 1626:1638 higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1626:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 0 34 theme PAP 81:83 arg1 O-glycosylation 57:71 altered O-glycosylation 49:71 altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers 49:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 6 35 theme expression 968:977 arg1 datasets 979:986 several prostate cancer gene expression datasets 939:986 several prostate cancer gene expression datasets 939:986 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 2 36 gly glycoprotein 402:413 arg1 glycoprotein 402:413 glycoprotein synthetic enzymes 402:431 glycoprotein synthetic enzymes 402:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 2 37 theme normal 441:446 arg1 tissues 471:477 normal and malignant prostate tissues 441:477 normal and malignant prostate tissues 441:477 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 2 38 theme synthetic 415:423 arg1 enzymes 425:431 glycoprotein synthetic enzymes 402:431 glycoprotein synthetic enzymes 402:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 11 39 theme new 1794:1796 arg1 avenue 1798:1803 a new avenue 1792:1803 a new avenue for development of prostate cancer specific glycoprotein biomarkers 1792:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 0 40 theme Increased 0:8 arg1 expression 10:19 Increased expression 0:19 Increased expression of GCNT1 0:28 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 1 41 theme prostate 281:288 arg1 cancer 290:295 prostate cancer 281:295 prostate cancer 281:295 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 8 42 link O-linked 1200:1207 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 2 43 theme prostate 462:469 arg1 tissues 471:477 normal and malignant prostate tissues 441:477 normal and malignant prostate tissues 441:477 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 11 44 from biology 1766:1772 arg1 important 1737:1745 important 1737:1745 important 1737:1745 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 11 45 theme cancer 1833:1838 arg1 biomarkers 1862:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 9 46 theme normal 1509:1514 arg1 samples 1539:1545 paired normal and malignant prostate samples 1502:1545 paired normal and malignant prostate samples 1502:1545 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 9 47 theme malignant 1520:1528 arg1 samples 1539:1545 paired normal and malignant prostate samples 1502:1545 paired normal and malignant prostate samples 1502:1545 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 8 48 theme near 1360:1363 arg1 differences 1377:1387 near significant differences 1360:1387 near significant differences in PAP sLe(x) levels (P = 0.06) 1360:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 3 49 theme expression 525:534 arg1 data 536:539 prostate cancer gene expression data 504:539 prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes 504:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 5 50 theme Mucin1 798:803 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 10 51 from cancer 1596:1601 arg1 over-expressed 1569:1582 over-expressed 1569:1582 over-expressed 1569:1582 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 52 theme MUC1 1675:1678 arg1 proteins 1680:1687 MUC1 proteins 1675:1687 MUC1 proteins 1675:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 11 53 gly glycoprotein 1849:1860 arg1 glycoprotein 1849:1860 prostate cancer specific glycoprotein biomarkers 1824:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 5 54 link O-linked 721:728 arg1 proteins 852:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 9 55 theme PAP 1436:1438 arg1 levels 1448:1453 PAP protein levels 1436:1453 PAP protein levels 1436:1453 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 4 56 theme prostate 657:664 arg1 samples 666:672 prostate samples 657:672 prostate samples using RT-PCR 657:685 Over-expression of GCNT1 was validated in prostate samples using RT-PCR. 24854630 11 57 theme prostate 1824:1831 arg1 biomarkers 1862:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 8 58 theme PAP 1392:1394 arg1 x 1400:1400 x 1400:1400 x 1400:1400 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 8 58 theme PAP 1392:1394 arg1 sLe 1396:1398 PAP sLe 1392:1398 PAP sLe(x) levels (P = 0.06) 1392:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 2 59 theme expression 381:390 arg1 levels 392:397 gene expression levels 376:397 gene expression levels of glycoprotein synthetic enzymes 376:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 8 60 theme PSA 1140:1142 arg1 PAP 1145:1147 PSA, PAP, and MUC1 capture antibodies 1140:1176 PAP 1145:1147 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 10 61 theme O-sLe 1650:1654 arg1 levels 1633:1638 higher levels 1626:1638 higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1626:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 8 62 from PSA 1341:1343 arg1 P = 0.06 1411:1418 P = 0.06 1411:1418 P = 0.06 1411:1418 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 8 62 from PSA 1341:1343 arg1 levels 1403:1408 PAP sLe(x) levels 1392:1408 PAP sLe(x) levels (P = 0.06) 1392:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 10 63 from levels 1633:1638 arg1 PSA 1662:1664 PSA 1662:1664 PSA 1662:1664 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 63 from levels 1633:1638 arg1 PAP 1667:1669 PAP 1667:1669 PAP 1667:1669 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 63 from levels 1633:1638 arg1 proteins 1680:1687 MUC1 proteins 1675:1687 MUC1 proteins 1675:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 4 64 theme GCNT1 634:638 arg1 Over-expression 615:629 Over-expression 615:629 Over-expression of GCNT1 615:638 Over-expression of GCNT1 was validated in prostate samples using RT-PCR. 24854630 5 65 theme core 714:717 arg1 proteins 852:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 9 66 theme prostate 1530:1537 arg1 samples 1539:1545 paired normal and malignant prostate samples 1502:1545 paired normal and malignant prostate samples 1502:1545 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 3 67 theme glycoprotein 583:594 arg1 enzymes 606:612 glycoprotein synthetic enzymes 583:612 glycoprotein synthetic enzymes 583:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 11 68 link O-linked 1705:1712 arg1 glycosylation 1714:1726 O-linked glycosylation 1705:1726 O-linked glycosylation 1705:1726 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 6 69 theme key 872:874 arg1 GCNT1 897:901 GCNT1 897:901 GCNT1 897:901 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 6 69 theme key 872:874 arg1 glycosyltransferase 876:894 A key glycosyltransferase 870:894 A key glycosyltransferase 870:894 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 11 70 theme prostate 1750:1757 arg1 biology 1766:1772 prostate cancer biology 1750:1772 prostate cancer biology 1750:1772 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 9 71 theme protein 1440:1446 arg1 levels 1448:1453 PAP protein levels 1436:1453 PAP protein levels 1436:1453 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 7 72 from tissue 1080:1085 arg1 samples 1119:1125 fresh-frozen prostate tissue samples 1090:1125 fresh-frozen prostate tissue samples 1090:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 10 73 theme core 1643:1646 arg1 O-sLe 1650:1654 core 2 O-sLe 1643:1654 core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1643:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 73 theme core 1643:1646 arg1 x 1656:1656 x 1656:1656 x 1656:1656 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 7 74 theme prostate 1103:1110 arg1 samples 1119:1125 fresh-frozen prostate tissue samples 1090:1125 fresh-frozen prostate tissue samples 1090:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 8 75 theme capture 1159:1165 arg1 antibodies 1167:1176 PSA, PAP, and MUC1 capture antibodies 1140:1176 antibodies 1167:1176 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 10 76 from O-sLe 1650:1654 arg1 PSA 1662:1664 PSA 1662:1664 PSA 1662:1664 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 76 from O-sLe 1650:1654 arg1 PAP 1667:1669 PAP 1667:1669 PAP 1667:1669 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 10 76 from O-sLe 1650:1654 arg1 proteins 1680:1687 MUC1 proteins 1675:1687 MUC1 proteins 1675:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 8 77 theme x 1213:1213 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 1 78 theme common 160:165 arg1 modification 185:196 a common posttranslational modification 158:196 a common posttranslational modification 158:196 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 78 theme common 160:165 arg1 glycosylation 141:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 6 79 theme gene 963:966 arg1 datasets 979:986 several prostate cancer gene expression datasets 939:986 several prostate cancer gene expression datasets 939:986 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 8 80 theme detection 1216:1224 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 11 81 theme O-linked 1705:1712 arg1 glycosylation 1714:1726 O-linked glycosylation 1705:1726 O-linked glycosylation 1705:1726 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 0 82 theme human 98:102 arg1 cancers 113:119 human prostate cancers 98:119 human prostate cancers 98:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 3 83 theme enzymes 606:612 arg1 expression 569:578 expression 569:578 expression of glycoprotein synthetic enzymes 569:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 8 84 theme specific 1184:1191 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 6 85 theme prostate 947:954 arg1 datasets 979:986 several prostate cancer gene expression datasets 939:986 several prostate cancer gene expression datasets 939:986 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 11 86 theme cancer 1759:1764 arg1 biology 1766:1772 prostate cancer biology 1750:1772 prostate cancer biology 1750:1772 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 5 87 theme acidic 827:832 arg1 PAP 847:849 PAP 847:849 PAP 847:849 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 87 theme acidic 827:832 arg1 phosphatase 834:844 prostatic acidic phosphatase 817:844 prostatic acidic phosphatase (PAP) 817:850 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 7 88 from levels 1027:1032 arg1 samples 1044:1050 cancer samples 1037:1050 cancer samples 1037:1050 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 10 89 theme CONCLUSIONS 1548:1558 arg1 GCNT1 1560:1564 CONCLUSIONS GCNT1 1548:1564 CONCLUSIONS GCNT1 1548:1564 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 5 90 theme O-linked 721:728 arg1 proteins 852:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 11 91 theme glycoprotein 1849:1860 arg1 biomarkers 1862:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 8 92 theme sLe 1209:1211 arg1 antibody 1226:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 a specific core 2 O-linked sLe(x) detection antibody 1182:1233 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 7 93 theme prostate 1071:1078 arg1 tissue 1080:1085 normal prostate tissue 1064:1085 normal prostate tissue in fresh-frozen prostate tissue samples 1064:1125 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 8 94 theme glycan 1266:1271 arg1 structure 1273:1281 this glycan structure 1261:1281 this glycan structure 1261:1281 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 5 95 theme sialyl 737:742 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 7 96 theme increased 1006:1014 arg1 levels 1027:1032 increased transcript levels 1006:1032 increased transcript levels in cancer samples 1006:1050 RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. 24854630 3 97 theme cancer 513:518 arg1 data 536:539 prostate cancer gene expression data 504:539 prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes 504:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 5 98 theme prostate 765:772 arg1 PSA 792:794 PSA 792:794 PSA 792:794 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 5 98 theme prostate 765:772 arg1 antigen 783:789 prostate specific antigen 765:789 prostate specific antigen (PSA) 765:795 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 10 99 from PSA 1662:1664 arg1 levels 1633:1638 higher levels 1626:1638 higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1626:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 11 100 theme specific 1840:1847 arg1 biomarkers 1862:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 prostate cancer specific glycoprotein biomarkers 1824:1871 Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers. 24854630 5 101 theme X 750:750 arg1 proteins 852:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 3 102 gly glycoprotein 583:594 arg1 glycoprotein 583:594 glycoprotein synthetic enzymes 583:612 glycoprotein synthetic enzymes 583:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 5 103 theme antigen 783:789 arg1 X 750:750 glycan sialyl Lewis X 730:750 core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins 714:859 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 1 104 theme Protein 133:139 arg1 modification 185:196 a common posttranslational modification 158:196 a common posttranslational modification 158:196 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 104 theme Protein 133:139 arg1 glycosylation 141:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation 122:153 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 10 105 from PAP 1667:1669 arg1 levels 1633:1638 higher levels 1626:1638 higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1626:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 8 106 theme normal 1305:1310 arg1 tissues 1312:1318 normal tissues 1305:1318 normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06) 1305:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 0 107 gly O-glycosylation 57:71 arg1 PSA 76:78 PSA 76:78 PSA 76:78 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 0 107 gly O-glycosylation 57:71 arg1 cancers 113:119 human prostate cancers 98:119 human prostate cancers 98:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 0 107 gly O-glycosylation 57:71 arg1 PAP 81:83 PAP 81:83 PAP 81:83 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 0 107 gly O-glycosylation 57:71 arg1 MUC1 90:93 MUC1 90:93 MUC1 90:93 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 10 108 theme higher 1626:1631 arg1 levels 1633:1638 higher levels 1626:1638 higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins 1626:1687 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 0 109 theme MUC1 90:93 arg1 O-glycosylation 57:71 altered O-glycosylation 49:71 altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers 49:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 2 110 theme glycoprotein 402:413 arg1 enzymes 425:431 glycoprotein synthetic enzymes 402:431 glycoprotein synthetic enzymes 402:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 0 111 theme GCNT1 24:28 arg1 expression 10:19 Increased expression 0:19 Increased expression of GCNT1 0:28 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 9 112 theme paired 1502:1507 arg1 samples 1539:1545 paired normal and malignant prostate samples 1502:1545 paired normal and malignant prostate samples 1502:1545 MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples. 24854630 2 113 theme malignant 452:460 arg1 tissues 471:477 normal and malignant prostate tissues 441:477 normal and malignant prostate tissues 441:477 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 3 114 theme prostate 504:511 arg1 data 536:539 prostate cancer gene expression data 504:539 prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes 504:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 1 115 theme glycan 202:207 arg1 changes 220:226 glycan structural changes 202:226 glycan structural changes 202:226 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 2 116 theme enzymes 425:431 arg1 levels 392:397 gene expression levels 376:397 gene expression levels of glycoprotein synthetic enzymes 376:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 24854630 0 117 theme altered 49:55 arg1 O-glycosylation 57:71 altered O-glycosylation 49:71 altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers 49:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 1 118 located observed 238:245 arg2 changes 220:226 glycan structural changes 202:226 glycan structural changes 202:226 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 118 located observed 238:245 arg1 malignancies 258:269 several malignancies 250:269 several malignancies including prostate cancer 250:295 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 1 118 located observed 238:245 arg1 cancer 290:295 prostate cancer 281:295 prostate cancer 281:295 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 10 119 from over-expressed 1569:1582 arg1 cancer 1596:1601 prostate cancer 1587:1601 prostate cancer 1587:1601 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 0 120 theme prostate 104:111 arg1 cancers 113:119 human prostate cancers 98:119 human prostate cancers 98:119 Increased expression of GCNT1 is associated with altered O-glycosylation of PSA, PAP, and MUC1 in human prostate cancers. 24854630 3 121 dep METHODS 480:486 arg1 We 488:489 We 488:489 We 488:489 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 6 122 dep RESULTS 862:868 arg1 GCNT1 897:901 GCNT1 897:901 GCNT1 897:901 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 6 122 dep RESULTS 862:868 arg1 glycosyltransferase 876:894 A key glycosyltransferase 870:894 A key glycosyltransferase 870:894 RESULTS A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. 24854630 8 123 theme significant 1365:1375 arg1 differences 1377:1387 near significant differences 1360:1387 near significant differences in PAP sLe(x) levels (P = 0.06) 1360:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 3 124 theme gene 520:523 arg1 data 536:539 prostate cancer gene expression data 504:539 prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes 504:612 METHODS We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. 24854630 8 125 from MUC1 1324:1327 arg1 P = 0.06 1411:1418 P = 0.06 1411:1418 P = 0.06 1411:1418 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 8 125 from MUC1 1324:1327 arg1 levels 1403:1408 PAP sLe(x) levels 1392:1408 PAP sLe(x) levels (P = 0.06) 1392:1419 ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). 24854630 5 126 used used 698:701 arg2 ELISA 688:692 ELISA 688:692 ELISA 688:692 ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins. 24854630 1 127 theme structural 209:218 arg1 changes 220:226 glycan structural changes 202:226 glycan structural changes 202:226 BACKGROUND Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. 24854630 10 128 theme prostate 1587:1594 arg1 cancer 1596:1601 prostate cancer 1587:1601 prostate cancer 1587:1601 CONCLUSIONS GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. 24854630 2 129 theme gene 376:379 arg1 levels 392:397 gene expression levels 376:397 gene expression levels of glycoprotein synthetic enzymes 376:431 We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues. 25279697 0 0 theme α-dystroglycan 81:94 arg1 glycosylation 64:76 the LARGE-dependent functional glycosylation 33:76 the LARGE-dependent functional glycosylation of α-dystroglycan 33:94 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 2 1 with responsible 426:436 arg1 structures 507:516 no established O-mannose initiated structures 472:516 no established O-mannose initiated structures containing a β3 linked GlcNAc known 472:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 5 2 theme secondary 1266:1274 arg1 dystroglycanopathies 1276:1295 secondary dystroglycanopathies 1266:1295 secondary dystroglycanopathies 1266:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 2 3 theme glycosylation 390:402 arg1 pathway 404:410 the O-mannosylation protein glycosylation pathway 362:410 the O-mannosylation protein glycosylation pathway 362:410 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 2 4 from defects 351:357 arg1 pathway 404:410 the O-mannosylation protein glycosylation pathway 362:410 the O-mannosylation protein glycosylation pathway 362:410 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 3 5 with β-1,3-N-acetylglucosaminyltransferase 640:676 arg1 activity 693:700 catalytic activity 683:700 catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose 683:821 Here we report this enzyme is not a β-1,3-N-acetylglucosaminyltransferase with catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose. 25279697 2 6 contain containing 518:527 arg2 GlcNAc 541:546 a β3 linked GlcNAc 529:546 a β3 linked GlcNAc known 529:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 2 6 contain containing 518:527 arg1 structures 507:516 no established O-mannose initiated structures 472:516 no established O-mannose initiated structures containing a β3 linked GlcNAc known 472:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 0 7 gly glycosylation 64:76 arg1 α-dystroglycan 81:94 α-dystroglycan 81:94 α-dystroglycan 81:94 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 2 8 theme human 590:594 arg1 enzyme 596:601 this human enzyme 585:601 this human enzyme 585:601 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 5 9 theme functional 1011:1020 arg1 structure 1044:1052 the functional O-mannosylated glycan structure 1007:1052 the functional O-mannosylated glycan structure 1007:1052 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 3 10 theme xylose 816:821 arg1 β-anomers 803:811 β-anomers 803:811 β-anomers 803:811 Here we report this enzyme is not a β-1,3-N-acetylglucosaminyltransferase with catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose. 25279697 3 10 theme xylose 816:821 arg1 α- 796:797 α- 796:797 α- 796:797 Here we report this enzyme is not a β-1,3-N-acetylglucosaminyltransferase with catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose. 25279697 2 11 link linked 534:539 arg1 GlcNAc 541:546 a β3 linked GlcNAc 529:546 a β3 linked GlcNAc known 529:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 1 12 theme α-dystroglycan 296:309 arg1 hypoglycosylation 275:291 hypoglycosylation 275:291 hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies) 275:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 4 13 theme priming 954:960 arg1 enzyme 962:967 the priming enzyme 950:967 the priming enzyme for LARGE 950:977 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 4 13 theme priming 954:960 arg1 B4GAT1 940:945 B4GAT1 940:945 B4GAT1 940:945 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 1 14 theme Recent 97:102 arg1 studies 104:110 Recent studies 97:110 Recent studies 97:110 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 0 15 theme priming 14:20 arg1 B4GAT1 0:5 B4GAT1 0:5 B4GAT1 0:5 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 0 15 theme priming 14:20 arg1 enzyme 22:27 the priming enzyme 10:27 the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan 10:94 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 4 16 theme LARGE 842:846 arg1 capable 858:864 capable 858:864 capable 858:864 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 4 16 theme LARGE 842:846 arg1 enzyme 848:853 The dual-activity LARGE enzyme 824:853 The dual-activity LARGE enzyme 824:853 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 5 17 gly O-mannosylated 1022:1035 arg1 structure 1044:1052 the functional O-mannosylated glycan structure 1007:1052 the functional O-mannosylated glycan structure 1007:1052 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 1 18 from mutations 130:138 arg1 B3GNT1 143:148 B3GNT1 143:148 B3GNT1 143:148 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 1 18 from mutations 130:138 arg1 enzyme 154:159 an enzyme 151:159 an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis 151:221 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 5 19 theme repeating 1132:1140 arg1 disaccharide 1142:1153 the LARGE-dependent repeating disaccharide 1112:1153 the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1112:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 5 19 theme repeating 1132:1140 arg1 necessary 1163:1171 necessary 1163:1171 necessary 1163:1171 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 2 20 theme O-mannose 487:495 arg1 structures 507:516 no established O-mannose initiated structures 472:516 no established O-mannose initiated structures containing a β3 linked GlcNAc known 472:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 2 21 theme initiated 497:505 arg1 structures 507:516 no established O-mannose initiated structures 472:516 no established O-mannose initiated structures containing a β3 linked GlcNAc known 472:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 1 22 theme secondary 312:320 arg1 α-dystroglycan 296:309 α-dystroglycan 296:309 α-dystroglycan (secondary dystroglycanopathies) 296:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 1 22 theme secondary 312:320 arg1 dystroglycanopathies 322:341 secondary dystroglycanopathies 312:341 secondary dystroglycanopathies 312:341 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 5 23 theme protein 1198:1204 arg1 binding 1206:1212 extracellular matrix protein binding 1177:1212 extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1177:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 2 24 theme linked 534:539 arg1 GlcNAc 541:546 a β3 linked GlcNAc 529:546 a β3 linked GlcNAc known 529:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 2 25 theme established 475:485 arg1 structures 507:516 no established O-mannose initiated structures 472:516 no established O-mannose initiated structures containing a β3 linked GlcNAc known 472:552 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 5 26 theme O-mannosylated 1022:1035 arg1 structure 1044:1052 the functional O-mannosylated glycan structure 1007:1052 the functional O-mannosylated glycan structure 1007:1052 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 3 27 theme catalytic 683:691 arg1 activity 693:700 catalytic activity 683:700 catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose 683:821 Here we report this enzyme is not a β-1,3-N-acetylglucosaminyltransferase with catalytic activity towards β-galactose but rather a β-1,4-glucuronyltransferase, designated B4GAT1, towards both α- and β-anomers of xylose. 25279697 4 28 theme experimental 913:924 arg1 evidence 926:933 experimental evidence that B4GAT1 is the priming enzyme for LARGE 913:977 experimental evidence that B4GAT1 is the priming enzyme for LARGE 913:977 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 0 29 theme LARGE-dependent 37:51 arg1 glycosylation 64:76 the LARGE-dependent functional glycosylation 33:76 the LARGE-dependent functional glycosylation of α-dystroglycan 33:94 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 5 30 theme O-mannosylated 1217:1230 arg1 α-dystroglycan 1232:1245 O-mannosylated α-dystroglycan 1217:1245 O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1217:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 2 31 theme protein 382:388 arg1 pathway 404:410 the O-mannosylation protein glycosylation pathway 362:410 the O-mannosylation protein glycosylation pathway 362:410 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 1 32 with dystrophy 260:268 arg1 hypoglycosylation 275:291 hypoglycosylation 275:291 hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies) 275:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 2 33 theme O-mannosylation 366:380 arg1 pathway 404:410 the O-mannosylation protein glycosylation pathway 362:410 the O-mannosylation protein glycosylation pathway 362:410 Since defects in the O-mannosylation protein glycosylation pathway are primarily responsible for dystroglycanopathies and with no established O-mannose initiated structures containing a β3 linked GlcNAc known, we biochemically interrogated this human enzyme. 25279697 1 34 gly hypoglycosylation 275:291 arg1 α-dystroglycan 296:309 α-dystroglycan 296:309 α-dystroglycan (secondary dystroglycanopathies) 296:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 1 34 gly hypoglycosylation 275:291 arg1 dystroglycanopathies 322:341 secondary dystroglycanopathies 312:341 secondary dystroglycanopathies 312:341 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 1 34 gly hypoglycosylation 275:291 arg1 dystrophy 260:268 congenital muscular dystrophy 240:268 congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies) 240:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 5 35 theme disaccharide 1142:1153 arg1 initiation 1098:1107 the initiation 1094:1107 the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1094:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 4 36 theme dual-activity 828:840 arg1 capable 858:864 capable 858:864 capable 858:864 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 4 36 theme dual-activity 828:840 arg1 enzyme 848:853 The dual-activity LARGE enzyme 824:853 The dual-activity LARGE enzyme 824:853 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 5 37 theme extracellular 1177:1189 arg1 matrix 1191:1196 extracellular matrix 1177:1196 extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1177:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 4 38 theme B4GAT1 891:896 arg1 products 879:886 products 879:886 products of B4GAT1 879:896 The dual-activity LARGE enzyme is capable of extending products of B4GAT1 and we provide experimental evidence that B4GAT1 is the priming enzyme for LARGE. 25279697 5 39 gly O-mannosylated 1217:1230 arg1 α-dystroglycan 1232:1245 O-mannosylated α-dystroglycan 1217:1245 O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1217:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 5 40 theme glycan 1037:1042 arg1 structure 1044:1052 the functional O-mannosylated glycan structure 1007:1052 the functional O-mannosylated glycan structure 1007:1052 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 5 41 theme matrix 1191:1196 arg1 binding 1206:1212 extracellular matrix protein binding 1177:1212 extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1177:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 1 42 theme poly-N-acetyllactosamine 188:211 arg1 synthesis 213:221 poly-N-acetyllactosamine synthesis 188:221 poly-N-acetyllactosamine synthesis 188:221 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 1 43 theme congenital 240:249 arg1 dystrophy 260:268 congenital muscular dystrophy 240:268 congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies) 240:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25279697 0 44 theme functional 53:62 arg1 glycosylation 64:76 the LARGE-dependent functional glycosylation 33:76 the LARGE-dependent functional glycosylation of α-dystroglycan 33:94 B4GAT1 is the priming enzyme for the LARGE-dependent functional glycosylation of α-dystroglycan. 25279697 5 45 theme LARGE-dependent 1116:1130 arg1 disaccharide 1142:1153 the LARGE-dependent repeating disaccharide 1112:1153 the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies 1112:1295 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 5 45 theme LARGE-dependent 1116:1130 arg1 necessary 1163:1171 necessary 1163:1171 necessary 1163:1171 Our results further define the functional O-mannosylated glycan structure and indicate that B4GAT1 is involved in the initiation of the LARGE-dependent repeating disaccharide that is necessary for extracellular matrix protein binding to O-mannosylated α-dystroglycan that is lacking in secondary dystroglycanopathies. 25279697 1 46 theme muscular 251:258 arg1 dystrophy 260:268 congenital muscular dystrophy 240:268 congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies) 240:342 Recent studies demonstrated that mutations in B3GNT1, an enzyme proposed to be involved in poly-N-acetyllactosamine synthesis, were causal for congenital muscular dystrophy with hypoglycosylation of α-dystroglycan (secondary dystroglycanopathies). 25318901 0 0 theme Escherichia 78:88 arg1 strain 100:105 Escherichia coli O174 strain 78:105 Escherichia coli O174 strain 78:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 1 theme O-antigen 150:158 arg1 unit 138:141 unit 138:141 unit of the O-antigen of Escherichia coli O174 strain 138:190 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 1 2 theme monosaccharide 270:283 arg1 intermediates 285:297 suitably functionalized monosaccharide intermediates 246:297 suitably functionalized monosaccharide intermediates 246:297 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 2 3 used used 406:409 arg2 Activation 300:309 Activation 300:309 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) 300:395 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 1 4 theme intermediates 285:297 arg1 glycosylations 228:241 sequential glycosylations 217:241 sequential glycosylations of suitably functionalized monosaccharide intermediates 217:297 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 0 5 theme O174 95:98 arg1 strain 100:105 Escherichia coli O174 strain 78:105 Escherichia coli O174 strain 78:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 6 gly glycosylations 228:241 arg1 intermediates 285:297 suitably functionalized monosaccharide intermediates 246:297 suitably functionalized monosaccharide intermediates 246:297 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 2 7 theme derivatives 344:354 arg1 Activation 300:309 Activation 300:309 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) 300:395 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 2 8 theme trichloroacetimidate 323:342 arg1 derivatives 344:354 glycosyl trichloroacetimidate derivatives 314:354 glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) 314:395 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 2 9 theme glycosyl 314:321 arg1 derivatives 344:354 glycosyl trichloroacetimidate derivatives 314:354 glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) 314:395 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 3 10 theme satisfactory 481:492 arg1 outcome 501:507 satisfactory stereo outcome 481:507 satisfactory stereo outcome 481:507 The glycosylation steps were high yielding with satisfactory stereo outcome. 25318901 1 11 theme Escherichia 163:173 arg1 strain 185:190 Escherichia coli O174 strain 163:190 Escherichia coli O174 strain 163:190 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 0 12 theme strain 100:105 arg1 O-antigen 65:73 the O-antigen 61:73 the O-antigen of Escherichia coli O174 strain 61:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 3 13 theme glycosylation 437:449 arg1 high 462:465 high 462:465 high 462:465 The glycosylation steps were high yielding with satisfactory stereo outcome. 25318901 3 13 theme glycosylation 437:449 arg1 steps 451:455 The glycosylation steps 433:455 The glycosylation steps 433:455 The glycosylation steps were high yielding with satisfactory stereo outcome. 25318901 0 14 dep Escherichia 78:88 arg1 coli 90:93 coli 90:93 coli 90:93 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 15 theme O174 180:183 arg1 strain 185:190 Escherichia coli O174 strain 163:190 Escherichia coli O174 strain 163:190 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 0 16 theme Efficient 0:8 arg1 synthesis 10:18 Efficient synthesis 0:18 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 0:106 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 17 theme strain 185:190 arg1 O-antigen 150:158 the O-antigen 146:158 the O-antigen of Escherichia coli O174 strain 146:190 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 2 18 theme nitrosyl 362:369 arg1 NOBF4 390:394 NOBF4 390:394 NOBF4 390:394 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 2 18 theme nitrosyl 362:369 arg1 tetrafluoroborate 371:387 nitrosyl tetrafluoroborate 362:387 nitrosyl tetrafluoroborate (NOBF4) 362:395 Activation of glycosyl trichloroacetimidate derivatives using nitrosyl tetrafluoroborate (NOBF4) has been used during the synthesis. 25318901 0 19 theme repeating 43:51 arg1 unit 53:56 the tetrasaccharide repeating unit 23:56 the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain 23:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 3 20 theme stereo 494:499 arg1 outcome 501:507 satisfactory stereo outcome 481:507 satisfactory stereo outcome 481:507 The glycosylation steps were high yielding with satisfactory stereo outcome. 25318901 0 21 theme tetrasaccharide 27:41 arg1 unit 53:56 the tetrasaccharide repeating unit 23:56 the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain 23:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 22 theme sequential 217:226 arg1 glycosylations 228:241 sequential glycosylations 217:241 sequential glycosylations of suitably functionalized monosaccharide intermediates 217:297 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 1 23 dep Escherichia 163:173 arg1 coli 175:178 coli 175:178 coli 175:178 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25318901 0 24 theme unit 53:56 arg1 synthesis 10:18 Efficient synthesis 0:18 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 0:106 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 0 25 theme O-antigen 65:73 arg1 unit 53:56 the tetrasaccharide repeating unit 23:56 the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain 23:105 Efficient synthesis of the tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain. 25318901 1 26 theme functionalized 255:268 arg1 intermediates 285:297 suitably functionalized monosaccharide intermediates 246:297 suitably functionalized monosaccharide intermediates 246:297 The tetrasaccharide repeating unit of the O-antigen of Escherichia coli O174 strain was synthesized applying sequential glycosylations of suitably functionalized monosaccharide intermediates. 25648510 0 0 theme sulfates 88:95 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 0 0 theme sulfates 88:95 arg1 sites 55:59 branching sites 45:59 branching sites of fucosylated chondroitin sulfates from sea cucumbers 45:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 9 1 from FCS 1645:1647 arg1 determination 1591:1603 the determination 1587:1603 the determination of a structure-activity relationship in FCS 1587:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 2 2 with oligosaccharides 485:500 arg1 structure 520:528 well-defined structure 507:528 well-defined structure related to various fragments of these polysaccharides 507:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 4 3 theme target 753:758 arg1 compounds 760:768 The target compounds 749:768 The target compounds 749:768 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 0 4 theme chondroitin 76:86 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 5 5 theme targets 952:958 arg1 units 929:933 the fucose units 918:933 the fucose units of the synthetic targets 918:958 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 7 6 theme remote 1236:1241 arg1 participation 1243:1255 the remote participation 1232:1255 the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side 1232:1407 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 8 7 theme such 1498:1501 arg1 participation 1503:1515 such participation 1498:1515 such participation 1498:1515 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 0 8 theme sea 102:104 arg1 cucumbers 106:114 sea cucumbers 102:114 sea cucumbers 102:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 5 9 theme FCS 1016:1018 arg1 structures 1020:1029 holothurian FCS structures 1004:1029 holothurian FCS structures 1004:1029 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 5 10 theme holothurian 1004:1014 arg1 structures 1020:1029 holothurian FCS structures 1004:1029 holothurian FCS structures 1004:1029 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 1 11 theme various 274:280 arg1 coagulation 318:328 blood coagulation 312:328 blood coagulation 312:328 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 thrombosis 331:340 thrombosis 331:340 thrombosis 331:340 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 bacterial 371:379 bacterial 371:379 bacterial 371:379 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 processes 293:301 various biological processes 274:301 various biological processes 274:301 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 angiogenesis 343:354 angiogenesis 343:354 angiogenesis 343:354 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 adhesion 391:398 viral adhesion 385:398 viral adhesion 385:398 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 11 theme various 274:280 arg1 inflammation 357:368 inflammation 357:368 inflammation 357:368 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 12 theme glycosyl 1321:1328 arg1 cations 1330:1336 the stabilized glycosyl cations 1306:1336 the stabilized glycosyl cations 1306:1336 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 0 13 from sites 55:59 arg1 cucumbers 106:114 sea cucumbers 102:114 sea cucumbers 102:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 6 14 theme α-glycoside 1047:1057 arg1 formation 1064:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. 25648510 0 15 gly fucosylated 64:74 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 1 16 theme biological 282:291 arg1 coagulation 318:328 blood coagulation 312:328 blood coagulation 312:328 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 thrombosis 331:340 thrombosis 331:340 thrombosis 331:340 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 bacterial 371:379 bacterial 371:379 bacterial 371:379 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 processes 293:301 various biological processes 274:301 various biological processes 274:301 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 angiogenesis 343:354 angiogenesis 343:354 angiogenesis 343:354 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 adhesion 391:398 viral adhesion 385:398 viral adhesion 385:398 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 16 theme biological 282:291 arg1 inflammation 357:368 inflammation 357:368 inflammation 357:368 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 17 theme stabilized 1310:1319 arg1 cations 1330:1336 the stabilized glycosyl cations 1306:1336 the stabilized glycosyl cations 1306:1336 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 7 18 theme glycosyl 1370:1377 arg1 acceptor 1379:1386 the glycosyl acceptor 1366:1386 the glycosyl acceptor only from the α-side 1366:1407 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 5 19 theme O-Sulfation 895:905 arg1 pattern 907:913 O-Sulfation pattern 895:913 O-Sulfation pattern in the fucose units of the synthetic targets 895:958 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 4 20 theme O-3 832:834 arg1 α-l-fucosyl 836:846 O-3 α-l-fucosyl 832:846 O-3 α-l-fucosyl 832:846 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 2 21 theme various 541:547 arg1 fragments 549:557 various fragments 541:557 various fragments of these polysaccharides 541:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 2 22 theme well-defined 507:518 arg1 structure 520:528 well-defined structure 507:528 well-defined structure related to various fragments of these polysaccharides 507:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 5 23 theme fucose 922:927 arg1 units 929:933 the fucose units 918:933 the fucose units of the synthetic targets 918:958 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 2 24 theme systematic 461:470 arg1 synthesis 472:480 systematic synthesis 461:480 systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides 461:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 1 25 theme viral 385:389 arg1 adhesion 391:398 viral adhesion 385:398 viral adhesion 385:398 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 26 theme glycosylation 1201:1213 arg1 outcome 1186:1192 Stereochemical outcome 1171:1192 Stereochemical outcome of the glycosylation 1171:1213 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 4 27 theme propyl 786:791 arg1 residue 813:819 propyl β-d-glucuronic acid residue 786:819 propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents 786:892 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 1 28 theme sea 193:195 arg1 cucumbers 197:205 sea cucumbers 193:205 sea cucumbers 193:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 29 theme Stereochemical 1171:1184 arg1 outcome 1186:1192 Stereochemical outcome 1171:1192 Stereochemical outcome of the glycosylation 1171:1213 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 6 30 theme Stereospecific 1032:1045 arg1 formation 1064:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. 25648510 2 31 dep started 453:459 arg1 determine 404:412 determine 404:412 To determine pharmacophore fragments in FCS 401:443 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 8 32 with agreement 1448:1456 arg1 energies 1486:1493 the SCF/MP2 calculated energies 1463:1493 the SCF/MP2 calculated energies of such participation 1463:1515 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 8 33 theme calculated 1475:1484 arg1 energies 1486:1493 the SCF/MP2 calculated energies 1463:1493 the SCF/MP2 calculated energies of such participation 1463:1515 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 4 34 dep α-l-fucosyl 836:846 arg1 substituents 881:892 -α-l-fucosyl substituents 868:892 -α-l-fucosyl substituents 868:892 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 0 35 theme oligosaccharides 17:32 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of the oligosaccharides 0:32 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 8 36 theme experimental 1414:1425 arg1 results 1427:1433 The experimental results 1410:1433 The experimental results 1410:1433 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 4 37 theme -α-l-fucosyl 868:879 arg1 substituents 881:892 -α-l-fucosyl substituents 868:892 -α-l-fucosyl substituents 868:892 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 4 38 theme acid 808:811 arg1 residue 813:819 propyl β-d-glucuronic acid residue 786:819 propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents 786:892 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 6 39 theme bond 1059:1062 arg1 formation 1064:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation 1032:1072 Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. 25648510 9 40 from determination 1591:1603 arg1 FCS 1645:1647 FCS 1645:1647 FCS 1645:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 7 41 with groups 1277:1282 arg1 formation 1293:1301 the formation 1289:1301 the formation 1289:1301 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 4 42 theme β-d-glucuronic 793:806 arg1 residue 813:819 propyl β-d-glucuronic acid residue 786:819 propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents 786:892 The target compounds are built up of propyl β-d-glucuronic acid residue bearing at O-3 α-l-fucosyl or α-l-fucosyl-(1→3)-α-l-fucosyl substituents. 25648510 9 43 theme model 1567:1571 arg1 compounds 1573:1581 model compounds 1567:1581 model compounds for the determination of a structure-activity relationship in FCS 1567:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 9 43 theme model 1567:1571 arg1 oligosaccharides 1534:1549 The synthesized oligosaccharides 1518:1549 The synthesized oligosaccharides 1518:1549 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 1 44 theme great 215:219 arg1 attention 221:229 great attention 215:229 great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion 215:398 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 45 theme groups 1277:1282 arg1 participation 1243:1255 the remote participation 1232:1255 the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side 1232:1407 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 8 46 theme good 1443:1446 arg1 agreement 1448:1456 good agreement 1443:1456 good agreement with the SCF/MP2 calculated energies of such participation 1443:1515 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 1 47 gly fucosylated 149:159 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 47 gly fucosylated 149:159 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 3 48 theme branching 712:720 arg1 FCS 731:733 FCS 731:733 FCS 731:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 3 48 theme branching 712:720 arg1 sites 722:726 branching sites 712:726 branching sites of FCS 712:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 9 49 theme structure-activity 1610:1627 arg1 relationship 1629:1640 a structure-activity relationship 1608:1640 a structure-activity relationship in FCS 1608:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 2 50 theme oligosaccharides 485:500 arg1 synthesis 472:480 systematic synthesis 461:480 systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides 461:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 1 51 theme blood 312:316 arg1 coagulation 318:328 blood coagulation 312:328 blood coagulation 312:328 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 2 52 theme polysaccharides 568:582 arg1 fragments 549:557 various fragments 541:557 various fragments of these polysaccharides 541:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 1 53 dep due 240:242 arg1 nowadays 231:238 nowadays 231:238 nowadays 231:238 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 5 54 from pattern 907:913 arg1 units 929:933 the fucose units 918:933 the fucose units of the synthetic targets 918:958 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 25648510 2 55 theme pharmacophore 414:426 arg1 fragments 428:436 pharmacophore fragments 414:436 pharmacophore fragments in FCS 414:443 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 7 56 theme cations 1330:1336 arg1 formation 1293:1301 the formation 1289:1301 the formation 1289:1301 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 3 57 theme related 701:707 arg1 trisaccharides 673:686 trisaccharides 673:686 trisaccharides structurally related to branching sites of FCS 673:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 9 58 from relationship 1629:1640 arg1 FCS 1645:1647 FCS 1645:1647 FCS 1645:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 1 59 theme Natural 117:123 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 59 theme Natural 117:123 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 0 60 theme branching 45:53 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 0 60 theme branching 45:53 arg1 sites 55:59 branching sites 45:59 branching sites of fucosylated chondroitin sulfates from sea cucumbers 45:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 3 61 theme FCS 731:733 arg1 FCS 731:733 FCS 731:733 FCS 731:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 3 61 theme FCS 731:733 arg1 sites 722:726 branching sites 712:726 branching sites of FCS 712:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 3 62 theme non-sulfated 625:636 arg1 di- 665:667 non-sulfated and selectively O-sulfated di- 625:667 non-sulfated and selectively O-sulfated di- 625:667 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 1 63 theme anionic 125:131 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 63 theme anionic 125:131 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 7 64 from α-side 1402:1407 arg1 acceptor 1379:1386 the glycosyl acceptor 1366:1386 the glycosyl acceptor only from the α-side 1366:1407 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 0 65 from cucumbers 106:114 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 0 65 from cucumbers 106:114 arg1 sites 55:59 branching sites 45:59 branching sites of fucosylated chondroitin sulfates from sea cucumbers 45:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 7 66 theme chloroacetyl 1264:1275 arg1 groups 1277:1282 the chloroacetyl groups 1260:1282 the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side 1260:1407 Stereochemical outcome of the glycosylation was explained by the remote participation of the chloroacetyl groups with the formation of the stabilized glycosyl cations, which could be attacked by the glycosyl acceptor only from the α-side. 25648510 1 67 theme polysaccharides 133:147 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 67 theme polysaccharides 133:147 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 8 68 theme participation 1503:1515 arg1 energies 1486:1493 the SCF/MP2 calculated energies 1463:1493 the SCF/MP2 calculated energies of such participation 1463:1515 The experimental results were in good agreement with the SCF/MP2 calculated energies of such participation. 25648510 1 69 theme due 240:242 arg1 attention 221:229 great attention 215:229 great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion 215:398 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 3 70 theme trisaccharides 673:686 arg1 synthesis 612:620 the synthesis 608:620 the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS 608:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 2 71 from fragments 428:436 arg1 FCS 441:443 FCS 441:443 FCS 441:443 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 1 72 theme fucosylated 149:159 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 72 theme fucosylated 149:159 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 6 73 theme 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl 1093:1136 arg1 donor 1164:1168 a donor 1162:1168 a donor 1162:1168 Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. 25648510 6 73 theme 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl 1093:1136 arg1 trichloroacetimidate 1138:1157 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate 1093:1157 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate 1093:1157 Stereospecific α-glycoside bond formation was achieved using 2-O-benzyl-3,4-di-O-chloroacetyl-α-l-fucosyl trichloroacetimidate as a donor. 25648510 0 74 theme fucosylated 64:74 arg1 sulfates 88:95 fucosylated chondroitin sulfates 64:95 fucosylated chondroitin sulfates from sea cucumbers 64:114 Synthesis of the oligosaccharides related to branching sites of fucosylated chondroitin sulfates from sea cucumbers. 25648510 9 75 theme relationship 1629:1640 arg1 determination 1591:1603 the determination 1587:1603 the determination of a structure-activity relationship in FCS 1587:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 3 76 theme O-sulfated 654:663 arg1 di- 665:667 non-sulfated and selectively O-sulfated di- 625:667 non-sulfated and selectively O-sulfated di- 625:667 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 1 77 from cucumbers 197:205 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 77 from cucumbers 197:205 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 78 theme chondroitin 161:171 arg1 sulfates 173:180 Natural anionic polysaccharides fucosylated chondroitin sulfates 117:180 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers 117:205 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 1 78 theme chondroitin 161:171 arg1 FCS 183:185 FCS 183:185 FCS 183:185 Natural anionic polysaccharides fucosylated chondroitin sulfates (FCS) from sea cucumbers attract great attention nowadays due to their ability to influence various biological processes, such as blood coagulation, thrombosis, angiogenesis, inflammation, bacterial and viral adhesion. 25648510 9 79 theme synthesized 1522:1532 arg1 compounds 1573:1581 model compounds 1567:1581 model compounds for the determination of a structure-activity relationship in FCS 1567:1647 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 9 79 theme synthesized 1522:1532 arg1 oligosaccharides 1534:1549 The synthesized oligosaccharides 1518:1549 The synthesized oligosaccharides 1518:1549 The synthesized oligosaccharides are regarded as model compounds for the determination of a structure-activity relationship in FCS. 25648510 2 80 theme related 530:536 arg1 structure 520:528 well-defined structure 507:528 well-defined structure related to various fragments of these polysaccharides 507:582 To determine pharmacophore fragments in FCS we have started systematic synthesis of oligosaccharides with well-defined structure related to various fragments of these polysaccharides. 25648510 3 81 theme di- 665:667 arg1 synthesis 612:620 the synthesis 608:620 the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS 608:733 In this communication, the synthesis of non-sulfated and selectively O-sulfated di- and trisaccharides structurally related to branching sites of FCS is described. 25648510 5 82 theme synthetic 942:950 arg1 targets 952:958 the synthetic targets 938:958 the synthetic targets 938:958 O-Sulfation pattern in the fucose units of the synthetic targets was selected according to the known to date holothurian FCS structures. 26630650 16 0 theme families 2248:2255 arg1 position 2230:2237 a more isolated position 2214:2237 a more isolated position of these families 2214:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 5 1 theme transcriptome 629:641 arg1 analysis 601:608 cDNA analysis 596:608 cDNA analysis of the venom gland transcriptome 596:641 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 4 2 attach isolated 484:491 arg1 venom 502:506 the venom 498:506 the venom of the spider Cupiennius salei 498:537 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 4 2 attach isolated 484:491 arg2 enzyme 473:478 a hyaluronidase-like enzyme 452:478 a hyaluronidase-like enzyme 452:478 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 10 3 theme heparan 1204:1210 arg1 sulfate 1212:1218 heparan sulfate 1204:1218 heparan sulfate 1204:1218 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 12 4 from end 1467:1469 arg1 Identification 1379:1392 Identification 1379:1392 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers 1379:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 12 4 from end 1467:1469 arg1 N-acetylglucosamine 1406:1424 terminal N-acetylglucosamine 1397:1424 terminal N-acetylglucosamine 1397:1424 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 12 4 from end 1467:1469 arg1 N-acetylgalactosamine 1429:1449 N-acetylgalactosamine 1429:1449 N-acetylgalactosamine 1429:1449 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 12 5 theme oligomers 1478:1486 arg1 end 1467:1469 the reducing end 1454:1469 the reducing end of the oligomers 1454:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 12 6 theme N-acetylglucosamine 1406:1424 arg1 Identification 1379:1392 Identification 1379:1392 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers 1379:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 14 7 theme neurotoxic 1788:1797 arg1 activity 1799:1806 the neurotoxic activity 1784:1806 the neurotoxic activity of CsTx-1 1784:1816 The enzyme significantly enhances the neurotoxic activity of CsTx-1. 26630650 8 8 theme M 984:984 arg1 KCl 986:988 0.2 M KCl 980:988 0.2 M KCl 980:988 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 12 9 theme N-acetylgalactosamine 1429:1449 arg1 Identification 1379:1392 Identification 1379:1392 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers 1379:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 15 10 from families 1980:1987 arg1 species 1964:1970 39 spider species 1954:1970 39 spider species from 21 families 1954:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 6 11 theme mass 781:784 arg1 spectrometry 786:797 tandem mass spectrometry 774:797 tandem mass spectrometry 774:797 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 13 12 theme enzyme 1671:1676 arg1 coinjection 1652:1662 coinjection 1652:1662 coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1652:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 12 13 theme reducing 1458:1465 arg1 end 1467:1469 the reducing end 1454:1469 the reducing end of the oligomers 1454:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 13 14 from effect 1574:1579 arg1 tissue 1630:1635 invertebrate tissue 1617:1635 invertebrate tissue 1617:1635 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 16 15 theme current 2185:2191 arg1 idea 2206:2209 the current phylogenetic idea 2181:2209 the current phylogenetic idea on a more isolated position of these families 2181:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 15 theme current 2185:2191 arg1 This 2116:2119 This 2116:2119 This 2116:2119 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 15 theme current 2185:2191 arg1 loss 2141:2144 a loss 2139:2144 a loss of this enzyme 2139:2159 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 5 16 theme venom 617:621 arg1 transcriptome 629:641 the venom gland transcriptome 613:641 the venom gland transcriptome 613:641 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 2 17 theme vertebrate 317:326 arg1 tissue 328:333 vertebrate tissue 317:333 vertebrate tissue 317:333 In several studies this spreading effect was tested on vertebrate tissue. 26630650 1 18 theme venom 198:202 arg1 STRUCTURE 119:127 STRUCTURE 119:127 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 119:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 1 18 theme venom 198:202 arg1 components 204:213 important venom components 188:213 important venom components acting as spreading factor of toxic compounds 188:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 11 19 theme chondroitin 1325:1335 arg1 A 1345:1345 chondroitin sulfate A 1325:1345 chondroitin sulfate A 1325:1345 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 0 20 theme salei 112:116 arg1 Venom 81:85 the Venom 77:85 the Venom of the Spider Cupiennius salei 77:116 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 15 21 with tests 1853:1857 arg1 hyaluronan 1864:1873 hyaluronan 1864:1873 hyaluronan 1864:1873 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 21 with tests 1853:1857 arg1 sulfate 1929:1935 heparan sulfate 1921:1935 heparan sulfate 1921:1935 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 21 with tests 1853:1857 arg1 A 1896:1896 chondroitin sulfate A 1876:1896 chondroitin sulfate A 1876:1896 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 21 with tests 1853:1857 arg1 sulfate 1908:1914 dermatan sulfate 1899:1914 dermatan sulfate 1899:1914 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 21 with tests 1853:1857 arg1 venoms 1942:1947 venoms 1942:1947 venoms from 39 spider species from 21 families 1942:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 16 22 theme catching 2306:2313 arg1 techniques 2315:2324 specialized prey catching techniques 2289:2324 specialized prey catching techniques 2289:2324 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 7 23 theme EGF-like 813:820 arg1 domain 822:827 A C-terminal EGF-like domain 800:827 A C-terminal EGF-like domain 800:827 A C-terminal EGF-like domain was identified in spider hyaluronidase using InterPro. 26630650 9 24 theme Divalent 991:998 arg1 ions 1000:1003 Divalent ions 991:1003 Divalent ions 991:1003 Divalent ions did not enhance HA degradation activity, indicating that they are not recruited for catalysis. 26630650 13 25 theme enzyme 1607:1612 arg1 effect 1574:1579 The spreading effect 1560:1579 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue 1560:1635 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 1 26 theme spreading 225:233 arg1 factor 235:240 spreading factor 225:240 spreading factor of toxic compounds 225:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 10 27 theme HYALURONIDASES 1118:1131 arg1 FUNCTION 1100:1107 FUNCTION 1100:1107 FUNCTION OF VENOM HYALURONIDASES 1100:1131 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 16 28 theme isolated 2221:2228 arg1 position 2230:2237 a more isolated position 2214:2237 a more isolated position of these families 2214:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 5 29 theme protein 660:666 arg1 analysis 668:675 protein analysis 660:675 protein analysis 660:675 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 13 30 theme invertebrate 1617:1628 arg1 tissue 1630:1635 invertebrate tissue 1617:1635 invertebrate tissue 1617:1635 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 5 31 theme amino 544:548 arg1 sequence 555:562 The amino acid sequence 540:562 The amino acid sequence of the enzyme 540:576 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 15 32 theme degradation 1841:1851 arg1 tests 1853:1857 Comparative substrate degradation tests 1819:1857 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families 1819:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 2 33 theme several 265:271 arg1 studies 273:279 several studies 265:279 several studies 265:279 In several studies this spreading effect was tested on vertebrate tissue. 26630650 12 34 from Identification 1379:1392 arg1 end 1467:1469 the reducing end 1454:1469 the reducing end of the oligomers 1454:1486 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 9 35 theme degradation 1024:1034 arg1 activity 1036:1043 HA degradation activity 1021:1043 HA degradation activity 1021:1043 Divalent ions did not enhance HA degradation activity, indicating that they are not recruited for catalysis. 26630650 0 36 theme Enzyme 65:70 arg1 N-glycosylations 11:26 N-glycosylations 11:26 N-glycosylations 11:26 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 0 36 theme Enzyme 65:70 arg1 Isolation 0:8 Isolation 0:8 Isolation 0:8 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 0 36 theme Enzyme 65:70 arg1 Function 32:39 Function 32:39 Function 32:39 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 1 37 theme SALEI 143:147 arg1 STRUCTURE 119:127 STRUCTURE 119:127 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 119:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 1 37 theme SALEI 143:147 arg1 components 204:213 important venom components 188:213 important venom components acting as spreading factor of toxic compounds 188:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 15 38 theme hyaluronidase-like 2088:2105 arg1 enzymes 2107:2113 hyaluronidase-like enzymes 2088:2113 hyaluronidase-like enzymes 2088:2113 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 1 39 theme HYALURONIDASE 155:167 arg1 Hyaluronidases 169:182 VENOM HYALURONIDASE Hyaluronidases 149:182 CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 132:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 3 40 theme spreading 360:368 arg1 activity 370:377 the spreading activity 356:377 the spreading activity on invertebrates 356:394 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 11 41 theme end 1263:1265 arg1 tetramers 1306:1314 tetramers 1306:1314 tetramers 1306:1314 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 11 41 theme end 1263:1265 arg1 products 1267:1274 The end products 1259:1274 The end products of hyaluronan degradation 1259:1300 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 15 42 theme chondroitin 1876:1886 arg1 sulfate 1888:1894 chondroitin sulfate 1876:1894 chondroitin sulfate A 1876:1896 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 6 43 link N-linked 690:697 arg1 glycans 699:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 4 44 theme hyaluronidase-like 454:471 arg1 enzyme 473:478 a hyaluronidase-like enzyme 452:478 a hyaluronidase-like enzyme 452:478 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 13 45 theme Drosophila 1732:1741 arg1 flies 1743:1747 Drosophila flies 1732:1747 Drosophila flies 1732:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 6 46 theme N-linked 690:697 arg1 glycans 699:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 0 47 theme Spider 94:99 arg1 salei 112:116 the Spider Cupiennius salei 90:116 the Spider Cupiennius salei 90:116 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 8 48 theme spider 888:893 arg1 enzyme 914:919 The spider hyaluronidase-like enzyme 884:919 The spider hyaluronidase-like enzyme 884:919 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 11 49 theme degradation 1290:1300 arg1 tetramers 1306:1314 tetramers 1306:1314 tetramers 1306:1314 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 11 49 theme degradation 1290:1300 arg1 products 1267:1274 The end products 1259:1274 The end products of hyaluronan degradation 1259:1300 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 15 50 theme sulfate 1888:1894 arg1 A 1896:1896 chondroitin sulfate A 1876:1896 chondroitin sulfate A 1876:1896 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 3 51 theme spiders 424:430 arg1 data 345:348 data 345:348 data about the spreading activity on invertebrates 345:394 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 3 51 theme spiders 424:430 arg1 organisms 411:419 the main prey organisms 397:419 the main prey organisms of spiders 397:430 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 13 52 theme main 1704:1707 arg1 CsTx-1 1720:1725 main neurotoxin CsTx-1 1704:1725 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 10 53 theme VENOM 1112:1116 arg1 HYALURONIDASES 1118:1131 VENOM HYALURONIDASES 1112:1131 VENOM HYALURONIDASES 1112:1131 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 8 54 theme maximal 928:934 arg1 activity 936:943 maximal activity 928:943 maximal activity 928:943 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 15 55 theme Comparative 1819:1829 arg1 tests 1853:1857 Comparative substrate degradation tests 1819:1857 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families 1819:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 13 56 with coinjection 1652:1662 arg1 salei 1698:1702 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 3 57 theme main 401:404 arg1 data 345:348 data 345:348 data about the spreading activity on invertebrates 345:394 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 3 57 theme main 401:404 arg1 organisms 411:419 the main prey organisms 397:419 the main prey organisms of spiders 397:430 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 16 58 theme prey 2301:2304 arg1 techniques 2315:2324 specialized prey catching techniques 2289:2324 specialized prey catching techniques 2289:2324 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 9 59 theme HA 1021:1022 arg1 activity 1036:1043 HA degradation activity 1021:1043 HA degradation activity 1021:1043 Divalent ions did not enhance HA degradation activity, indicating that they are not recruited for catalysis. 26630650 4 60 theme Cupiennius 522:531 arg1 salei 533:537 the spider Cupiennius salei 511:537 the spider Cupiennius salei 511:537 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 13 61 theme Cupiennius 1687:1696 arg1 salei 1698:1702 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 6 62 theme hyaluronidase 725:737 arg1 glycosylations 739:752 honey bee hyaluronidase glycosylations 715:752 honey bee hyaluronidase glycosylations 715:752 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 15 63 dep families 2012:2019 arg1 Nephilidae 2056:2065 Nephilidae 2056:2065 Nephilidae 2056:2065 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 63 dep families 2012:2019 arg1 families 2012:2019 some spider families 2000:2019 some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) 2000:2066 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 63 dep families 2012:2019 arg1 Araneidae 2042:2050 Araneidae 2042:2050 Araneidae 2042:2050 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 63 dep families 2012:2019 arg1 Eresidae 2032:2039 Eresidae 2032:2039 Eresidae 2032:2039 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 6 64 theme honey 715:719 arg1 bee 721:723 honey bee 715:723 honey bee hyaluronidase glycosylations 715:752 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 10 65 theme chondroitin 1173:1183 arg1 sulfate 1185:1191 chondroitin sulfate 1173:1191 chondroitin sulfate A 1173:1193 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 11 66 dep tetramers 1306:1314 arg1 whereas 1317:1323 whereas 1317:1323 whereas 1317:1323 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 16 67 theme phylogenetic 2193:2204 arg1 idea 2206:2209 the current phylogenetic idea 2181:2209 the current phylogenetic idea on a more isolated position of these families 2181:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 67 theme phylogenetic 2193:2204 arg1 This 2116:2119 This 2116:2119 This 2116:2119 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 67 theme phylogenetic 2193:2204 arg1 loss 2141:2144 a loss 2139:2144 a loss of this enzyme 2139:2159 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 15 68 theme heparan 1921:1927 arg1 sulfate 1929:1935 heparan sulfate 1921:1935 heparan sulfate 1921:1935 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 5 69 theme gland 623:627 arg1 transcriptome 629:641 the venom gland transcriptome 613:641 the venom gland transcriptome 613:641 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 6 70 theme akin 707:710 arg1 glycans 699:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 14 71 theme CsTx-1 1811:1816 arg1 activity 1799:1806 the neurotoxic activity 1784:1806 the neurotoxic activity of CsTx-1 1784:1816 The enzyme significantly enhances the neurotoxic activity of CsTx-1. 26630650 8 72 theme 0.2 980:982 arg1 M 984:984 M 984:984 M 984:984 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 15 73 from species 1964:1970 arg1 venoms 1942:1947 venoms 1942:1947 venoms from 39 spider species from 21 families 1942:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 13 74 dep salei 1698:1702 arg1 CsTx-1 1720:1725 main neurotoxin CsTx-1 1704:1725 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 12 75 theme terminal 1397:1404 arg1 N-acetylglucosamine 1406:1424 terminal N-acetylglucosamine 1397:1424 terminal N-acetylglucosamine 1397:1424 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 15 76 theme dermatan 1899:1906 arg1 sulfate 1908:1914 dermatan sulfate 1899:1914 dermatan sulfate 1899:1914 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 2 77 theme spreading 286:294 arg1 effect 296:301 this spreading effect 281:301 this spreading effect 281:301 In several studies this spreading effect was tested on vertebrate tissue. 26630650 11 78 theme sulfate 1337:1343 arg1 A 1345:1345 chondroitin sulfate A 1325:1345 chondroitin sulfate A 1325:1345 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 6 79 theme tandem 774:779 arg1 spectrometry 786:797 tandem mass spectrometry 774:797 tandem mass spectrometry 774:797 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 7 80 theme spider 847:852 arg1 hyaluronidase 854:866 spider hyaluronidase 847:866 spider hyaluronidase using InterPro 847:881 A C-terminal EGF-like domain was identified in spider hyaluronidase using InterPro. 26630650 1 81 theme important 188:196 arg1 STRUCTURE 119:127 STRUCTURE 119:127 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 119:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 1 81 theme important 188:196 arg1 components 204:213 important venom components 188:213 important venom components acting as spreading factor of toxic compounds 188:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 0 82 from Venom 81:85 arg1 N-glycosylations 11:26 N-glycosylations 11:26 N-glycosylations 11:26 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 0 82 from Venom 81:85 arg1 Isolation 0:8 Isolation 0:8 Isolation 0:8 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 0 82 from Venom 81:85 arg1 Function 32:39 Function 32:39 Function 32:39 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 15 83 theme spider 1957:1962 arg1 species 1964:1970 39 spider species 1954:1970 39 spider species from 21 families 1954:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 13 84 theme hyaluronidase-like 1588:1605 arg1 enzyme 1607:1612 the hyaluronidase-like enzyme 1584:1612 the hyaluronidase-like enzyme 1584:1612 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 3 85 from activity 370:377 arg1 invertebrates 382:394 invertebrates 382:394 invertebrates 382:394 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 15 86 theme substrate 1831:1839 arg1 tests 1853:1857 Comparative substrate degradation tests 1819:1857 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families 1819:1987 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 13 87 theme spreading 1564:1572 arg1 effect 1574:1579 The spreading effect 1560:1579 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue 1560:1635 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 5 88 theme acid 550:553 arg1 sequence 555:562 The amino acid sequence 540:562 The amino acid sequence of the enzyme 540:576 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 10 89 theme dermatan 1224:1231 arg1 sulfate 1233:1239 dermatan sulfate 1224:1239 dermatan sulfate 1224:1239 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 1 90 theme CUPIENNIUS 132:141 arg1 SALEI 143:147 CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 132:182 CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 132:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 16 91 from idea 2206:2209 arg1 position 2230:2237 a more isolated position 2214:2237 a more isolated position of these families 2214:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 1 92 theme VENOM 149:153 arg1 Hyaluronidases 169:182 VENOM HYALURONIDASE Hyaluronidases 149:182 CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 132:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 11 93 theme hyaluronan 1279:1288 arg1 degradation 1290:1300 hyaluronan degradation 1279:1300 hyaluronan degradation 1279:1300 The end products of hyaluronan degradation are tetramers, whereas chondroitin sulfate A is mainly degraded to hexamers. 26630650 16 94 theme enzyme 2154:2159 arg1 This 2116:2119 This 2116:2119 This 2116:2119 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 94 theme enzyme 2154:2159 arg1 idea 2206:2209 the current phylogenetic idea 2181:2209 the current phylogenetic idea on a more isolated position of these families 2181:2255 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 94 theme enzyme 2154:2159 arg1 loss 2141:2144 a loss 2139:2144 a loss of this enzyme 2139:2159 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 16 95 theme specialized 2289:2299 arg1 techniques 2315:2324 specialized prey catching techniques 2289:2324 specialized prey catching techniques 2289:2324 This is interpreted as a loss of this enzyme and fits quite well the current phylogenetic idea on a more isolated position of these families and can perhaps be explained by specialized prey catching techniques. 26630650 0 96 theme Cupiennius 101:110 arg1 salei 112:116 the Spider Cupiennius salei 90:116 the Spider Cupiennius salei 90:116 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 5 97 theme cDNA 596:599 arg1 analysis 601:608 cDNA analysis 596:608 cDNA analysis of the venom gland transcriptome 596:641 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 6 98 theme complex 682:688 arg1 glycans 699:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans 678:705 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 15 99 theme spider 2005:2010 arg1 families 2012:2019 some spider families 2000:2019 some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) 2000:2066 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 99 theme spider 2005:2010 arg1 Nephilidae 2056:2065 Nephilidae 2056:2065 Nephilidae 2056:2065 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 99 theme spider 2005:2010 arg1 Araneidae 2042:2050 Araneidae 2042:2050 Araneidae 2042:2050 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 15 99 theme spider 2005:2010 arg1 Eresidae 2032:2039 Eresidae 2032:2039 Eresidae 2032:2039 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 7 100 theme C-terminal 802:811 arg1 domain 822:827 A C-terminal EGF-like domain 800:827 A C-terminal EGF-like domain 800:827 A C-terminal EGF-like domain was identified in spider hyaluronidase using InterPro. 26630650 5 101 theme enzyme 571:576 arg1 sequence 555:562 The amino acid sequence 540:562 The amino acid sequence of the enzyme 540:576 The amino acid sequence of the enzyme was determined by cDNA analysis of the venom gland transcriptome and confirmed by protein analysis. 26630650 15 102 theme enzymes 2107:2113 arg1 activity 2076:2083 activity 2076:2083 activity of hyaluronidase-like enzymes 2076:2113 Comparative substrate degradation tests with hyaluronan, chondroitin sulfate A, dermatan sulfate, and heparan sulfate with venoms from 39 spider species from 21 families identified some spider families (Atypidae, Eresidae, Araneidae and Nephilidae) without activity of hyaluronidase-like enzymes. 26630650 8 103 theme hyaluronidase-like 895:912 arg1 enzyme 914:919 The spider hyaluronidase-like enzyme 884:919 The spider hyaluronidase-like enzyme 884:919 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 13 104 theme neurotoxin 1709:1718 arg1 CsTx-1 1720:1725 main neurotoxin CsTx-1 1704:1725 the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies 1683:1747 The spreading effect of the hyaluronidase-like enzyme on invertebrate tissue was studied by coinjection of the enzyme with the Cupiennius salei main neurotoxin CsTx-1 into Drosophila flies. 26630650 1 105 dep SALEI 143:147 arg1 Hyaluronidases 169:182 VENOM HYALURONIDASE Hyaluronidases 149:182 CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases 132:182 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 12 106 theme endo-β-N-acetyl-D-hexosaminidase 1516:1547 arg1 hydrolase 1549:1557 an endo-β-N-acetyl-D-hexosaminidase hydrolase 1513:1557 an endo-β-N-acetyl-D-hexosaminidase hydrolase 1513:1557 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 12 106 theme endo-β-N-acetyl-D-hexosaminidase 1516:1547 arg1 enzyme 1503:1508 the enzyme 1499:1508 the enzyme 1499:1508 Identification of terminal N-acetylglucosamine or N-acetylgalactosamine at the reducing end of the oligomers identified the enzyme as an endo-β-N-acetyl-D-hexosaminidase hydrolase. 26630650 4 107 theme spider 515:520 arg1 salei 533:537 the spider Cupiennius salei 511:537 the spider Cupiennius salei 511:537 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 1 108 theme toxic 245:249 arg1 compounds 251:259 toxic compounds 245:259 toxic compounds 245:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 26630650 0 109 theme Hyaluronidase-Like 46:63 arg1 Enzyme 65:70 a Hyaluronidase-Like Enzyme 44:70 a Hyaluronidase-Like Enzyme 44:70 Isolation, N-glycosylations and Function of a Hyaluronidase-Like Enzyme from the Venom of the Spider Cupiennius salei. 26630650 3 110 theme prey 406:409 arg1 data 345:348 data 345:348 data about the spreading activity on invertebrates 345:394 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 3 110 theme prey 406:409 arg1 organisms 411:419 the main prey organisms 397:419 the main prey organisms of spiders 397:430 However, data about the spreading activity on invertebrates, the main prey organisms of spiders, are lacking. 26630650 4 111 theme salei 533:537 arg1 venom 502:506 the venom 498:506 the venom of the spider Cupiennius salei 498:537 Here, a hyaluronidase-like enzyme was isolated from the venom of the spider Cupiennius salei. 26630650 8 112 theme acidic 948:953 arg1 pH 955:956 acidic pH 948:956 acidic pH 948:956 The spider hyaluronidase-like enzyme showed maximal activity at acidic pH, between 40-60°C, and 0.2 M KCl. 26630650 6 113 theme bee 721:723 arg1 glycosylations 739:752 honey bee hyaluronidase glycosylations 715:752 honey bee hyaluronidase glycosylations 715:752 Two complex N-linked glycans akin to honey bee hyaluronidase glycosylations, were identified by tandem mass spectrometry. 26630650 10 114 theme sulfate 1185:1191 arg1 A 1193:1193 chondroitin sulfate A 1173:1193 chondroitin sulfate A 1173:1193 FUNCTION OF VENOM HYALURONIDASES Besides hyaluronan, the enzyme degrades chondroitin sulfate A, whereas heparan sulfate and dermatan sulfate are not affected. 26630650 1 115 theme compounds 251:259 arg1 factor 235:240 spreading factor 225:240 spreading factor of toxic compounds 225:259 STRUCTURE OF CUPIENNIUS SALEI VENOM HYALURONIDASE Hyaluronidases are important venom components acting as spreading factor of toxic compounds. 29029079 4 0 theme glycan 546:551 arg1 structures 553:562 glycan structures 546:562 glycan structures on native EC-SOD purified from human sera 546:604 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 4 1 theme biantennary 633:643 arg1 structures 645:654 sialylated biantennary structures 622:654 sialylated biantennary structures 622:654 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 5 2 theme furin-mediated 831:844 arg1 cleavage 857:864 furin-mediated C-terminal cleavage 831:864 furin-mediated C-terminal cleavage of EC-SOD 831:874 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 4 3 theme native 567:572 arg1 EC-SOD 574:579 native EC-SOD 567:579 native EC-SOD purified from human sera 567:604 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 4 4 theme sialylated 622:631 arg1 structures 645:654 sialylated biantennary structures 622:654 sialylated biantennary structures 622:654 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 4 5 from structures 553:562 arg1 EC-SOD 574:579 native EC-SOD 567:579 native EC-SOD purified from human sera 567:604 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 5 6 theme glycan 663:668 arg1 cells 702:706 glycan maturation-defective CHO mutant cells 663:706 glycan maturation-defective CHO mutant cells 663:706 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 2 7 theme posttranslational 311:327 arg1 N-glycosylation 353:367 N-glycosylation 353:367 N-glycosylation 353:367 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 2 7 theme posttranslational 311:327 arg1 cleavage 385:392 proteolytic cleavage 373:392 proteolytic cleavage 373:392 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 2 7 theme posttranslational 311:327 arg1 modifications 329:341 several posttranslational modifications 303:341 several posttranslational modifications including N-glycosylation and proteolytic cleavage 303:392 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 3 8 theme proteolytic 414:424 arg1 cleavage 426:433 proteolytic cleavage 414:433 proteolytic cleavage 414:433 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 5 9 theme maturation-defective 670:689 arg1 cells 702:706 glycan maturation-defective CHO mutant cells 663:706 glycan maturation-defective CHO mutant cells 663:706 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 2 10 theme several 303:309 arg1 N-glycosylation 353:367 N-glycosylation 353:367 N-glycosylation 353:367 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 2 10 theme several 303:309 arg1 cleavage 385:392 proteolytic cleavage 373:392 proteolytic cleavage 373:392 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 2 10 theme several 303:309 arg1 modifications 329:341 several posttranslational modifications 303:341 several posttranslational modifications including N-glycosylation and proteolytic cleavage 303:392 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 5 11 theme mutant 695:700 arg1 cells 702:706 glycan maturation-defective CHO mutant cells 663:706 glycan maturation-defective CHO mutant cells 663:706 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 5 12 theme CHO 691:693 arg1 cells 702:706 glycan maturation-defective CHO mutant cells 663:706 glycan maturation-defective CHO mutant cells 663:706 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 1 13 dep dismutase 132:140 arg1 EC-SOD 143:148 EC-SOD 143:148 EC-SOD 143:148 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 1 13 dep dismutase 132:140 arg1 SOD3 151:154 SOD3 151:154 SOD3 151:154 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 4 14 gly sialylated 622:631 arg1 structures 645:654 sialylated biantennary structures 622:654 sialylated biantennary structures 622:654 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 1 15 theme oxidative 182:190 arg1 damage 192:197 oxidative damage 182:197 oxidative damage 182:197 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 6 16 theme EC-SOD 960:965 arg1 modifications 943:955 the posttranslational modifications 921:955 the posttranslational modifications of EC-SOD 921:965 These results provide new insights into how the posttranslational modifications of EC-SOD control its functions. 29029079 3 17 dep structure 463:471 arg1 the 459:461 the 459:461 the 459:461 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 3 18 theme cleavage 426:433 arg1 roles 405:409 the roles 401:409 the roles of proteolytic cleavage 401:433 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 5 19 theme acids 766:770 arg1 presence 738:745 the presence 734:745 the presence of terminal sialic acids in the N-glycans of EC-SOD 734:797 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 1 20 theme Extracellular 107:119 arg1 dismutase 132:140 Extracellular superoxide dismutase 107:140 Extracellular superoxide dismutase (EC-SOD, SOD3) 107:155 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 5 21 attach presence 738:745 arg2 acids 766:770 terminal sialic acids 750:770 terminal sialic acids 750:770 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 5 21 attach presence 738:745 arg1 N-glycans 779:787 the N-glycans 775:787 the N-glycans of EC-SOD 775:797 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 0 22 theme extracellular 15:27 arg1 EC-SOD 51:56 EC-SOD 51:56 EC-SOD 51:56 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 0 22 theme extracellular 15:27 arg1 dismutase 40:48 extracellular superoxide dismutase 15:48 extracellular superoxide dismutase (EC-SOD) 15:57 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 1 23 theme superoxide 121:130 arg1 dismutase 132:140 Extracellular superoxide dismutase 107:140 Extracellular superoxide dismutase (EC-SOD, SOD3) 107:155 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 1 24 theme superoxide 214:223 arg1 anions 225:230 superoxide anions 214:230 superoxide anions 214:230 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 5 25 theme C-terminal 846:855 arg1 cleavage 857:864 furin-mediated C-terminal cleavage 831:864 furin-mediated C-terminal cleavage of EC-SOD 831:874 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 3 26 theme N-glycans 496:504 arg1 function 477:484 function 477:484 function 477:484 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 3 26 theme N-glycans 496:504 arg1 structure 463:471 structure 463:471 structure 463:471 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 0 27 theme dismutase 40:48 arg1 Sialylation 0:10 Sialylation 0:10 Sialylation of extracellular superoxide dismutase (EC-SOD) 0:57 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 1 28 theme cardiovascular 263:276 arg1 system 278:283 cardiovascular system 263:283 cardiovascular system 263:283 Extracellular superoxide dismutase (EC-SOD, SOD3) protects tissues against oxidative damage by detoxifying superoxide anions, particularly in the lungs and cardiovascular system. 29029079 6 29 theme posttranslational 925:941 arg1 modifications 943:955 the posttranslational modifications 921:955 the posttranslational modifications of EC-SOD 921:965 These results provide new insights into how the posttranslational modifications of EC-SOD control its functions. 29029079 0 30 theme superoxide 29:38 arg1 EC-SOD 51:56 EC-SOD 51:56 EC-SOD 51:56 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 0 30 theme superoxide 29:38 arg1 dismutase 40:48 extracellular superoxide dismutase 15:48 extracellular superoxide dismutase (EC-SOD) 15:57 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 5 31 theme EC-SOD 869:874 arg1 secretion 817:825 the secretion 813:825 the secretion 813:825 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 5 31 theme EC-SOD 869:874 arg1 cleavage 857:864 furin-mediated C-terminal cleavage 831:864 furin-mediated C-terminal cleavage of EC-SOD 831:874 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 4 32 theme human 595:599 arg1 sera 601:604 human sera 595:604 human sera 595:604 Here we analyzed glycan structures on native EC-SOD purified from human sera, and identified sialylated biantennary structures. 29029079 5 33 theme terminal 750:757 arg1 acids 766:770 terminal sialic acids 750:770 terminal sialic acids 750:770 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 5 34 theme sialic 759:764 arg1 acids 766:770 terminal sialic acids 750:770 terminal sialic acids 750:770 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 2 35 theme proteolytic 373:383 arg1 cleavage 385:392 proteolytic cleavage 373:392 proteolytic cleavage 373:392 EC-SOD undergoes several posttranslational modifications including N-glycosylation and proteolytic cleavage. 29029079 0 36 theme furin-mediated 68:81 arg1 cleavage 83:90 furin-mediated cleavage 68:90 furin-mediated cleavage 68:90 Sialylation of extracellular superoxide dismutase (EC-SOD) enhances furin-mediated cleavage and secretion. 29029079 6 37 theme new 899:901 arg1 insights 903:910 new insights 899:910 new insights into how the posttranslational modifications of EC-SOD control its functions 899:987 These results provide new insights into how the posttranslational modifications of EC-SOD control its functions. 29029079 3 38 theme EC-SOD 489:494 arg1 N-glycans 496:504 EC-SOD N-glycans 489:504 EC-SOD N-glycans 489:504 While the roles of proteolytic cleavage have been well studied, the structure and function of EC-SOD N-glycans are poorly understood. 29029079 5 39 from presence 738:745 arg1 N-glycans 779:787 the N-glycans 775:787 the N-glycans of EC-SOD 775:797 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 29029079 5 40 theme EC-SOD 792:797 arg1 N-glycans 779:787 the N-glycans 775:787 the N-glycans of EC-SOD 775:797 Using glycan maturation-defective CHO mutant cells, we further revealed that the presence of terminal sialic acids in the N-glycans of EC-SOD enhanced both the secretion and furin-mediated C-terminal cleavage of EC-SOD. 25210040 2 0 theme high 665:668 arg1 determinants 681:692 high stringency determinants 665:692 high stringency determinants 665:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 8 1 theme determinant 1760:1770 arg1 features 1726:1733 The structural features 1711:1733 The structural features of the LacdiNAc-inducing determinant on human TFF2 1711:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 2 theme extracellular 262:274 arg1 glycoproteins 283:295 extracellular matrix glycoproteins 262:295 extracellular matrix glycoproteins 262:295 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 8 3 theme candidate 1947:1955 arg1 proteins 1957:1964 novel candidate proteins 1941:1964 novel candidate proteins in the secretome of HEK-293 cells 1941:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 8 4 theme human 1775:1779 arg1 TFF2 1781:1784 human TFF2 1775:1784 human TFF2 1775:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 5 theme glycoproteins 283:295 arg1 number 252:257 a limited number 242:257 a limited number of extracellular matrix glycoproteins and neuropeptide hormones 242:321 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 0 6 theme Trefoil 0:6 arg1 domains 22:28 Trefoil factor family domains 0:28 Trefoil factor family domains 0:28 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 2 7 theme single 626:631 arg1 TFF 633:635 single TFF 626:635 single TFF (Trefoil factor) domains as high stringency determinants 626:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 2 7 theme single 626:631 arg1 factor 646:651 Trefoil factor 638:651 Trefoil factor 638:651 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 5 8 from LacdiNAc 1255:1262 arg1 chains 1293:1298 bi-antennary complex-type chains 1267:1298 bi-antennary complex-type chains 1267:1298 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 2 9 theme transfer 614:621 arg1 control 596:602 an entirely independent conformational control 557:602 an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants 557:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 5 10 theme TFF 1325:1327 arg1 domain 1329:1334 a single TFF domain 1316:1334 a single TFF domain 1316:1334 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 5 10 theme TFF 1325:1327 arg1 sufficient 1340:1349 sufficient 1340:1349 sufficient 1340:1349 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 8 11 theme crystal 1816:1822 arg1 structures 1824:1833 crystal structures 1816:1833 crystal structures of porcine TFF2 1816:1849 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 5 12 theme domain 1200:1205 arg1 probes 1207:1212 the recombinant full-length and single domain probes 1161:1212 the recombinant full-length and single domain probes 1161:1212 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 3 13 theme P1 839:840 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 13 theme P1 839:840 arg1 probes 855:860 single P1 or P2 domain probes 832:860 single P1 or P2 domain probes 832:860 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 1 14 located found 233:237 arg2 diamine 192:198 N'-di-N-acetyllactose diamine 170:198 N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) 170:228 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 1 14 located found 233:237 arg2 N 168:168 The disaccharide N 151:168 The disaccharide N 151:168 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 1 14 located found 233:237 arg1 number 252:257 a limited number 242:257 a limited number of extracellular matrix glycoproteins and neuropeptide hormones 242:321 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 3 15 theme Human 695:699 arg1 series 868:873 a series 866:873 a series of Cys/Gly mutant forms with aberrant domain structures 866:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 15 theme Human 695:699 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 15 theme Human 695:699 arg1 probes 855:860 single P1 or P2 domain probes 832:860 single P1 or P2 domain probes 832:860 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 15 theme Human 695:699 arg1 probe 962:966 a double point-mutated probe 939:966 a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch 939:1031 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 15 theme Human 695:699 arg1 probe 778:782 a wild type full-length probe 754:782 a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2) 754:826 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 16 theme TFF2 1846:1849 arg1 structures 1824:1833 crystal structures 1816:1833 crystal structures of porcine TFF2 1816:1849 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 7 17 theme determinant 1646:1656 arg1 components 1658:1667 essential determinant components 1636:1667 essential determinant components 1636:1667 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 17 theme determinant 1646:1656 arg1 these 1626:1630 these 1626:1630 these 1626:1630 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 5 18 theme fucosylated 1243:1253 arg1 LacdiNAc 1255:1262 fucosylated LacdiNAc 1243:1262 fucosylated LacdiNAc on bi-antennary complex-type chains 1243:1298 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 5 19 theme natural 1139:1145 arg1 TFF2 1155:1158 natural gastric TFF2 1139:1158 natural gastric TFF2 1139:1158 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 2 20 theme controlling 492:502 arg1 elements 504:511 controlling elements 492:511 controlling elements in LacdiNAc biosynthesis 492:536 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 3 21 theme forms 893:897 arg1 series 868:873 a series 866:873 a series of Cys/Gly mutant forms with aberrant domain structures 866:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 21 theme forms 893:897 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 0 22 theme N-linked 89:96 arg1 synthesis 140:148 N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis 89:148 synthesis 140:148 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 3 23 with forms 893:897 arg1 structures 920:929 aberrant domain structures 904:929 aberrant domain structures 904:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 1 24 theme N'-di-N-acetyllactose 170:190 arg1 N 168:168 The disaccharide N 151:168 The disaccharide N 151:168 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 1 24 theme N'-di-N-acetyllactose 170:190 arg1 diamine 192:198 N'-di-N-acetyllactose diamine 170:198 N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) 170:228 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 3 25 theme Cys/Gly 878:884 arg1 forms 893:897 Cys/Gly mutant forms 878:897 Cys/Gly mutant forms with aberrant domain structures 878:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 26 from features 1726:1733 arg1 TFF2 1781:1784 human TFF2 1775:1784 human TFF2 1775:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 5 27 theme single 1318:1323 arg1 domain 1329:1334 a single TFF domain 1316:1334 a single TFF domain 1316:1334 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 5 27 theme single 1318:1323 arg1 sufficient 1340:1349 sufficient 1340:1349 sufficient 1340:1349 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 3 28 theme P2 845:846 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 28 theme P2 845:846 arg1 probes 855:860 single P1 or P2 domain probes 832:860 single P1 or P2 domain probes 832:860 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 29 theme extracellular 1868:1880 arg1 glycoproteins 1915:1927 extracellular matrix-related LacdiNAc-positive glycoproteins 1868:1927 extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1868:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 2 30 theme conformational 581:594 arg1 control 596:602 an entirely independent conformational control 557:602 an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants 557:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 5 31 theme modification 1378:1389 arg1 transfer 1361:1368 transfer 1361:1368 transfer of this modification 1361:1389 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 8 32 theme LacdiNAc-positive 1897:1913 arg1 glycoproteins 1915:1927 extracellular matrix-related LacdiNAc-positive glycoproteins 1868:1927 extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1868:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 3 33 theme aberrant 904:911 arg1 structures 920:929 aberrant domain structures 904:929 aberrant domain structures 904:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 2 34 theme LacdiNAc 516:523 arg1 biosynthesis 525:536 LacdiNAc biosynthesis 516:536 LacdiNAc biosynthesis 516:536 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 8 35 from determinant 1760:1770 arg1 TFF2 1781:1784 human TFF2 1775:1784 human TFF2 1775:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 6 36 theme LacNAc 1498:1503 arg1 expression 1505:1514 preponderant LacNAc expression 1485:1514 preponderant LacNAc expression 1485:1514 Contrasting to this, the Cys/Gly mutants showed strongly reduced LacdiNAc levels and instead preponderant LacNAc expression. 25210040 5 37 theme single 1193:1198 arg1 probes 1207:1212 the recombinant full-length and single domain probes 1161:1212 the recombinant full-length and single domain probes 1161:1212 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 1 38 gly glycoproteins 283:295 arg1 glycoproteins 283:295 extracellular matrix glycoproteins 262:295 extracellular matrix glycoproteins 262:295 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 5 39 gly fucosylated 1243:1253 arg1 LacdiNAc 1255:1262 fucosylated LacdiNAc 1243:1262 fucosylated LacdiNAc on bi-antennary complex-type chains 1243:1298 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 0 40 theme family 15:20 arg1 domains 22:28 Trefoil factor family domains 0:28 Trefoil factor family domains 0:28 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 1 41 theme hormones 314:321 arg1 number 252:257 a limited number 242:257 a limited number of extracellular matrix glycoproteins and neuropeptide hormones 242:321 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 2 42 theme peptide 468:474 arg1 determinants 476:487 peptide determinants 468:487 peptide determinants as controlling elements in LacdiNAc biosynthesis 468:536 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 3 43 dep probe 778:782 arg1 containing 794:803 containing 794:803 containing TFF domains P1 and P2 794:825 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 44 gly glycoproteins 1915:1927 arg1 glycoproteins 1915:1927 extracellular matrix-related LacdiNAc-positive glycoproteins 1868:1927 extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1868:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 45 dep diamine 192:198 arg1 LacdiNAc 201:208 LacdiNAc 201:208 LacdiNAc 201:208 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 1 45 dep diamine 192:198 arg1 GalNAcβ1-4GlcNAcβ 211:227 GalNAcβ1-4GlcNAcβ 211:227 GalNAcβ1-4GlcNAcβ 211:227 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 6 46 theme reduced 1449:1455 arg1 levels 1466:1471 strongly reduced LacdiNAc levels 1440:1471 strongly reduced LacdiNAc levels 1440:1471 Contrasting to this, the Cys/Gly mutants showed strongly reduced LacdiNAc levels and instead preponderant LacNAc expression. 25210040 5 47 theme full-length 1177:1187 arg1 probes 1207:1212 the recombinant full-length and single domain probes 1161:1212 the recombinant full-length and single domain probes 1161:1212 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 3 48 dep containing 794:803 arg1 TFF2-Fl 785:791 TFF2-Fl 785:791 TFF2-Fl 785:791 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 7 49 theme aromatic 1572:1579 arg1 residues 1581:1588 two highly conserved aromatic residues 1551:1588 two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1551:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 3 50 theme point-mutated 948:960 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 50 theme point-mutated 948:960 arg1 T68Q/F59Q 969:977 T68Q/F59Q 969:977 T68Q/F59Q 969:977 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 50 theme point-mutated 948:960 arg1 probe 962:966 a double point-mutated probe 939:966 a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch 939:1031 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 51 from TFF2 1781:1784 arg1 features 1726:1733 The structural features 1711:1733 The structural features of the LacdiNAc-inducing determinant on human TFF2 1711:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 3 52 theme aromatic 988:995 arg1 residues 997:1004 aromatic residues 988:1004 aromatic residues within a hydrophobic patch 988:1031 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 53 theme HEK-293 737:743 arg1 cells 745:749 HEK-293 cells 737:749 HEK-293 cells 737:749 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 2 54 theme previous 424:431 arg1 studies 433:439 previous studies 424:439 previous studies 424:439 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 8 55 theme structural 1715:1724 arg1 features 1726:1733 The structural features 1711:1733 The structural features of the LacdiNAc-inducing determinant on human TFF2 1711:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 5 56 theme complex-type 1280:1291 arg1 chains 1293:1298 bi-antennary complex-type chains 1267:1298 bi-antennary complex-type chains 1267:1298 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 0 57 theme conformational 57:70 arg1 determinants 72:83 highly efficient conformational determinants 40:83 highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis 40:148 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 5 58 theme recombinant 1165:1175 arg1 probes 1207:1212 the recombinant full-length and single domain probes 1161:1212 the recombinant full-length and single domain probes 1161:1212 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 3 59 theme full-length 766:776 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 59 theme full-length 766:776 arg1 probe 778:782 a wild type full-length probe 754:782 a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2) 754:826 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 60 theme novel 1941:1945 arg1 proteins 1957:1964 novel candidate proteins 1941:1964 novel candidate proteins in the secretome of HEK-293 cells 1941:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 7 61 from residues 1581:1588 arg1 loop 1593:1596 loop 3 1593:1598 loop 3 (T68Q/F59Q) 1593:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 61 from residues 1581:1588 arg1 T68Q/F59Q 1601:1609 T68Q/F59Q 1601:1609 T68Q/F59Q 1601:1609 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 62 theme point 1532:1536 arg1 mutations 1538:1546 point mutations 1532:1546 point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1532:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 3 63 theme hydrophobic 1015:1025 arg1 patch 1027:1031 a hydrophobic patch 1013:1031 a hydrophobic patch 1013:1031 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 4 64 theme mass 1078:1081 arg1 spectrometry 1083:1094 mass spectrometry 1078:1094 mass spectrometry 1078:1094 The N-glycosylation probes were analyzed by mass spectrometry for their glycoprofiles. 25210040 3 65 theme wild 756:759 arg1 type 761:764 a wild type 754:764 a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2) 754:826 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 66 theme LacdiNAc-inducing 1742:1758 arg1 determinant 1760:1770 the LacdiNAc-inducing determinant 1738:1770 the LacdiNAc-inducing determinant on human TFF2 1738:1784 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 67 theme matrix 276:281 arg1 glycoproteins 283:295 extracellular matrix glycoproteins 262:295 extracellular matrix glycoproteins 262:295 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 7 68 from loop 1593:1596 arg1 mutations 1538:1546 point mutations 1532:1546 point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1532:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 0 69 theme efficient 47:55 arg1 determinants 72:83 highly efficient conformational determinants 40:83 highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis 40:148 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 6 70 theme Cys/Gly 1417:1423 arg1 mutants 1425:1431 the Cys/Gly mutants 1413:1431 the Cys/Gly mutants 1413:1431 Contrasting to this, the Cys/Gly mutants showed strongly reduced LacdiNAc levels and instead preponderant LacNAc expression. 25210040 3 71 dep domains 809:815 arg1 domains 809:815 TFF domains P1 and P2 805:825 TFF domains P1 and P2 805:825 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 71 dep domains 809:815 arg1 P2 824:825 P2 824:825 P2 824:825 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 71 dep domains 809:815 arg1 P1 817:818 P1 817:818 P1 817:818 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 7 72 theme LacdiNAc 1690:1697 arg1 expression 1699:1708 LacdiNAc expression 1690:1708 LacdiNAc expression 1690:1708 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 2 73 theme stringency 670:679 arg1 determinants 681:692 high stringency determinants 665:692 high stringency determinants 665:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 8 74 theme cells 1994:1998 arg1 secretome 1973:1981 the secretome 1969:1981 the secretome of HEK-293 cells 1969:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 3 75 theme TFF 805:807 arg1 domains 809:815 TFF domains P1 and P2 805:825 TFF domains P1 and P2 805:825 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 75 theme TFF 805:807 arg1 P2 824:825 P2 824:825 P2 824:825 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 75 theme TFF 805:807 arg1 P1 817:818 P1 817:818 P1 817:818 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 7 76 from mutations 1538:1546 arg1 loop 1593:1596 loop 3 1593:1598 loop 3 (T68Q/F59Q) 1593:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 76 from mutations 1538:1546 arg1 T68Q/F59Q 1601:1609 T68Q/F59Q 1601:1609 T68Q/F59Q 1601:1609 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 2 77 theme GalNAc 607:612 arg1 transfer 614:621 GalNAc transfer 607:621 GalNAc transfer 607:621 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 2 78 theme Trefoil 638:644 arg1 TFF 633:635 single TFF 626:635 single TFF (Trefoil factor) domains as high stringency determinants 626:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 2 78 theme Trefoil 638:644 arg1 factor 646:651 Trefoil factor 638:651 Trefoil factor 638:651 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 5 79 with agreement 1124:1132 arg1 TFF2 1155:1158 natural gastric TFF2 1139:1158 natural gastric TFF2 1139:1158 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 8 80 theme structures 1824:1833 arg1 series 1858:1863 a series 1856:1863 a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1856:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 8 80 theme structures 1824:1833 arg1 basis 1807:1811 the basis 1803:1811 the basis of crystal structures of porcine TFF2 1803:1849 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 7 81 theme essential 1636:1644 arg1 components 1658:1667 essential determinant components 1636:1667 essential determinant components 1636:1667 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 81 theme essential 1636:1644 arg1 these 1626:1630 these 1626:1630 these 1626:1630 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 2 82 theme TFF 633:635 arg1 domains 654:660 single TFF (Trefoil factor) domains 626:660 single TFF (Trefoil factor) domains as high stringency determinants 626:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 8 83 theme porcine 1838:1844 arg1 TFF2 1846:1849 porcine TFF2 1838:1849 porcine TFF2 1838:1849 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 84 theme disaccharide 155:166 arg1 N 168:168 The disaccharide N 151:168 The disaccharide N 151:168 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 1 84 theme disaccharide 155:166 arg1 diamine 192:198 N'-di-N-acetyllactose diamine 170:198 N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) 170:228 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 2 85 from elements 504:511 arg1 biosynthesis 525:536 LacdiNAc biosynthesis 516:536 LacdiNAc biosynthesis 516:536 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 3 86 theme single 832:837 arg1 P1 839:840 single P1 832:840 single P1 832:840 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 0 87 theme N 98:98 arg1 synthesis 140:148 N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis 89:148 synthesis 140:148 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 5 88 theme gastric 1147:1153 arg1 TFF2 1155:1158 natural gastric TFF2 1139:1158 natural gastric TFF2 1139:1158 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 0 89 dep synthesis 140:148 arg1 N'-di-N-acetyllactosediamine 100:127 N'-di-N-acetyllactosediamine 100:127 N'-di-N-acetyllactosediamine (LacdiNAc) 100:138 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 0 89 dep synthesis 140:148 arg1 LacdiNAc 130:137 LacdiNAc 130:137 LacdiNAc 130:137 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 8 90 theme HEK-293 1986:1992 arg1 cells 1994:1998 HEK-293 cells 1986:1998 HEK-293 cells 1986:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 1 91 theme glycosyltransferases 379:398 arg1 β4GalNAc-T3/T4 400:413 the glycosyltransferases β4GalNAc-T3/T4 375:413 the glycosyltransferases β4GalNAc-T3/T4 375:413 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 3 92 theme mutant 886:891 arg1 forms 893:897 Cys/Gly mutant forms 878:897 Cys/Gly mutant forms with aberrant domain structures 878:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 1 93 theme GalNAc 365:370 arg1 transfer 353:360 a protein-specific transfer 334:360 a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4 334:413 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 3 94 theme domain 848:853 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 94 theme domain 848:853 arg1 probes 855:860 single P1 or P2 domain probes 832:860 single P1 or P2 domain probes 832:860 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 2 95 theme independent 569:579 arg1 control 596:602 an entirely independent conformational control 557:602 an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants 557:692 Whereas previous studies have revealed evidence for peptide determinants as controlling elements in LacdiNAc biosynthesis, we report here on an entirely independent conformational control of GalNAc transfer by single TFF (Trefoil factor) domains as high stringency determinants. 25210040 0 96 theme factor 8:13 arg1 domains 22:28 Trefoil factor family domains 0:28 Trefoil factor family domains 0:28 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 8 97 theme matrix-related 1882:1895 arg1 glycoproteins 1915:1927 extracellular matrix-related LacdiNAc-positive glycoproteins 1868:1927 extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1868:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 3 98 theme domain 913:918 arg1 structures 920:929 aberrant domain structures 904:929 aberrant domain structures 904:929 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 8 99 from proteins 1957:1964 arg1 secretome 1973:1981 the secretome 1969:1981 the secretome of HEK-293 cells 1969:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 6 100 theme preponderant 1485:1496 arg1 expression 1505:1514 preponderant LacNAc expression 1485:1514 preponderant LacNAc expression 1485:1514 Contrasting to this, the Cys/Gly mutants showed strongly reduced LacdiNAc levels and instead preponderant LacNAc expression. 25210040 1 101 theme neuropeptide 301:312 arg1 hormones 314:321 neuropeptide hormones 301:321 neuropeptide hormones 301:321 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 8 102 theme glycoproteins 1915:1927 arg1 series 1858:1863 a series 1856:1863 a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells 1856:1998 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 8 102 theme glycoproteins 1915:1927 arg1 basis 1807:1811 the basis 1803:1811 the basis of crystal structures of porcine TFF2 1803:1849 The structural features of the LacdiNAc-inducing determinant on human TFF2 are discussed on the basis of crystal structures of porcine TFF2, and a series of extracellular matrix-related LacdiNAc-positive glycoproteins detected as novel candidate proteins in the secretome of HEK-293 cells. 25210040 3 103 theme double 941:946 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 103 theme double 941:946 arg1 T68Q/F59Q 969:977 T68Q/F59Q 969:977 T68Q/F59Q 969:977 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 103 theme double 941:946 arg1 probe 962:966 a double point-mutated probe 939:966 a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch 939:1031 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 4 104 theme N-glycosylation 1038:1052 arg1 probes 1054:1059 The N-glycosylation probes 1034:1059 The N-glycosylation probes 1034:1059 The N-glycosylation probes were analyzed by mass spectrometry for their glycoprofiles. 25210040 1 105 theme limited 244:250 arg1 number 252:257 a limited number 242:257 a limited number of extracellular matrix glycoproteins and neuropeptide hormones 242:321 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 6 106 theme LacdiNAc 1457:1464 arg1 levels 1466:1471 strongly reduced LacdiNAc levels 1440:1471 strongly reduced LacdiNAc levels 1440:1471 Contrasting to this, the Cys/Gly mutants showed strongly reduced LacdiNAc levels and instead preponderant LacNAc expression. 25210040 1 107 theme protein-specific 336:351 arg1 transfer 353:360 a protein-specific transfer 334:360 a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4 334:413 The disaccharide N,N'-di-N-acetyllactose diamine (LacdiNAc, GalNAcβ1-4GlcNAcβ) is found in a limited number of extracellular matrix glycoproteins and neuropeptide hormones indicating a protein-specific transfer of GalNAc by the glycosyltransferases β4GalNAc-T3/T4. 25210040 0 108 link N-linked 89:96 arg1 synthesis 140:148 N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis 89:148 synthesis 140:148 Trefoil factor family domains represent highly efficient conformational determinants for N-linked N,N'-di-N-acetyllactosediamine (LacdiNAc) synthesis. 25210040 7 109 theme residues 1581:1588 arg1 mutations 1538:1546 point mutations 1532:1546 point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1532:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 7 110 with probe 1521:1525 arg1 mutations 1538:1546 point mutations 1532:1546 point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1532:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 5 111 theme bi-antennary 1267:1278 arg1 chains 1293:1298 bi-antennary complex-type chains 1267:1298 bi-antennary complex-type chains 1267:1298 In agreement with natural gastric TFF2, the recombinant full-length and single domain probes expressed nearly exclusively fucosylated LacdiNAc on bi-antennary complex-type chains indicating that a single TFF domain was sufficient to induce transfer of this modification. 25210040 7 112 theme conserved 1562:1570 arg1 residues 1581:1588 two highly conserved aromatic residues 1551:1588 two highly conserved aromatic residues in loop 3 (T68Q/F59Q) 1551:1610 The probe with point mutations of two highly conserved aromatic residues in loop 3 (T68Q/F59Q) revealed that these are essential determinant components, as the probe lacked LacdiNAc expression. 25210040 3 113 theme type 761:764 arg1 TFF2 701:704 Human TFF2 695:704 Human TFF2 695:704 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 25210040 3 113 theme type 761:764 arg1 probe 778:782 a wild type full-length probe 754:782 a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2) 754:826 Human TFF2 was recombinantly expressed in HEK-293 cells as a wild type full-length probe (TFF2-Fl, containing TFF domains P1 and P2), as single P1 or P2 domain probes, as a series of Cys/Gly mutant forms with aberrant domain structures, and as a double point-mutated probe (T68Q/F59Q) lacking aromatic residues within a hydrophobic patch. 27558752 7 0 theme stress-responsive 1430:1446 arg1 degradation 1448:1458 the stress-responsive degradation 1426:1458 the stress-responsive degradation of high-mannose type N-glycans 1426:1489 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 4 1 theme glycoproteins 975:987 arg1 quantification 957:970 quantification 957:970 quantification 957:970 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 1 theme glycoproteins 975:987 arg1 identification 938:951 comprehensive identification 924:951 comprehensive identification 924:951 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 2 2 theme surface 402:408 arg1 topology 410:417 surface topology 402:417 surface topology 402:417 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 0 3 theme chilling 103:110 arg1 stress 112:117 chilling stress 103:117 chilling stress in Arabidopsis 103:132 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 4 4 from glycoproteins 975:987 arg1 mixture 1030:1036 a complex mixture 1020:1036 a complex mixture 1020:1036 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 6 5 theme molecular 1241:1249 arg1 signature 1251:1259 a molecular signature 1239:1259 a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature 1239:1347 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 7 6 theme Structural 1350:1359 arg1 N-glycoproteomics 1361:1377 Structural N-glycoproteomics 1350:1377 Structural N-glycoproteomics following time-course cold treatments 1350:1415 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 1 7 contain has 151:153 arg2 impact 163:168 a great impact 155:168 a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity 155:267 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 7 contain has 151:153 arg1 N-glycosylation 135:149 N-glycosylation 135:149 N-glycosylation 135:149 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 4 8 theme heterogeneous 993:1005 arg1 N-glycans 1007:1015 heterogeneous N-glycans 993:1015 heterogeneous N-glycans 993:1015 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 3 9 theme automated 659:667 arg1 searching 678:686 automated database searching 659:686 automated database searching 659:686 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 10 theme glycopeptides 610:622 arg1 searching 678:686 automated database searching 659:686 automated database searching 659:686 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 10 theme glycopeptides 610:622 arg1 labeling 576:583 (15) N-metabolic labeling 559:583 (15) N-metabolic labeling 559:583 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 10 theme glycopeptides 610:622 arg1 enrichment 596:605 selective enrichment 586:605 selective enrichment of glycopeptides 586:622 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 10 theme glycopeptides 610:622 arg1 analysis 646:653 data-dependent MS/MS analysis 625:653 data-dependent MS/MS analysis 625:653 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 4 11 theme database 846:853 arg1 size 855:858 the public database size 835:858 the public database size 835:858 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 7 12 theme relocation 1593:1602 arg1 degradation 1644:1654 degradation 1644:1654 degradation 1644:1654 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 12 theme relocation 1593:1602 arg1 misfolding 1629:1638 misfolding 1629:1638 misfolding 1629:1638 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 12 theme relocation 1593:1602 arg1 trafficking 1616:1626 trafficking 1616:1626 trafficking 1616:1626 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 12 theme relocation 1593:1602 arg1 transport 1605:1613 transport 1605:1613 transport 1605:1613 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 12 theme relocation 1593:1602 arg1 mechanisms 1571:1580 the cellular mechanisms 1558:1580 the cellular mechanisms of protein relocation 1558:1602 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 4 13 from quantification 957:970 arg1 mixture 1030:1036 a complex mixture 1020:1036 a complex mixture 1020:1036 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 7 14 theme time-course 1389:1399 arg1 treatments 1406:1415 time-course cold treatments 1389:1415 time-course cold treatments 1389:1415 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 3 15 theme N-metabolic 564:574 arg1 labeling 576:583 (15) N-metabolic labeling 559:583 (15) N-metabolic labeling 559:583 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 7 16 theme cellular 1562:1569 arg1 degradation 1644:1654 degradation 1644:1654 degradation 1644:1654 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 16 theme cellular 1562:1569 arg1 misfolding 1629:1638 misfolding 1629:1638 misfolding 1629:1638 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 16 theme cellular 1562:1569 arg1 trafficking 1616:1626 trafficking 1616:1626 trafficking 1616:1626 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 16 theme cellular 1562:1569 arg1 transport 1605:1613 transport 1605:1613 transport 1605:1613 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 16 theme cellular 1562:1569 arg1 mechanisms 1571:1580 the cellular mechanisms 1558:1580 the cellular mechanisms of protein relocation 1558:1602 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 6 17 theme specific 1261:1268 arg1 signature 1251:1259 a molecular signature 1239:1259 a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature 1239:1347 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 5 18 theme endoplasmic 1125:1135 arg1 reticulum 1137:1145 endoplasmic reticulum 1125:1145 the endoplasmic reticulum (ER) resident proteins 1121:1168 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 5 18 theme endoplasmic 1125:1135 arg1 ER 1148:1149 ER 1148:1149 ER 1148:1149 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 4 19 gly glycoproteins 723:735 arg1 glycoproteins 723:735 Arabidopsis glycoproteins 711:735 Arabidopsis glycoproteins 711:735 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 6 20 theme stress 1323:1328 arg1 degree 1307:1312 the degree 1303:1312 the degree of plant stress 1303:1328 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 5 21 theme Proteome-wide 1039:1051 arg1 analysis 1053:1060 Proteome-wide analysis 1039:1060 Proteome-wide analysis 1039:1060 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 3 22 theme proteomics 451:460 arg1 method 462:467 a highly sensitive proteomics method 432:467 a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins 432:534 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 4 23 from identification 938:951 arg1 mixture 1030:1036 a complex mixture 1020:1036 a complex mixture 1020:1036 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 24 theme comprehensive 924:936 arg1 identification 938:951 comprehensive identification 924:951 comprehensive identification 924:951 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 6 25 theme plant 1317:1321 arg1 stress 1323:1328 plant stress 1317:1328 plant stress 1317:1328 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 4 26 gly glycoproteins 975:987 arg1 glycoproteins 975:987 glycoproteins 975:987 glycoproteins 975:987 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 0 27 theme reticulum-associated 12:31 arg1 degradation 42:52 Endoplasmic reticulum-associated N-glycan degradation 0:52 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 0:133 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 3 28 theme selective 586:594 arg1 enrichment 596:605 selective enrichment 586:605 selective enrichment of glycopeptides 586:622 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 4 29 gly glycopeptides 741:753 arg2 glycopeptides 741:753 glycopeptides 741:753 glycopeptides 741:753 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 1 30 from impact 163:168 arg1 stability 211:219 stability 211:219 stability 211:219 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 30 from impact 163:168 arg1 activity 260:267 enzyme activity 253:267 enzyme activity 253:267 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 30 from impact 163:168 arg1 structure 186:194 glycoprotein structure 173:194 glycoprotein structure 173:194 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 30 from impact 163:168 arg1 solubility 222:231 solubility 222:231 solubility 222:231 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 30 from impact 163:168 arg1 immunogenicity 234:247 immunogenicity 234:247 immunogenicity 234:247 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 1 30 from impact 163:168 arg1 conformation 197:208 conformation 197:208 conformation 197:208 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 4 31 theme public 839:844 arg1 database 846:853 the public database 835:853 the public database size 835:858 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 7 32 theme high-mannose 1463:1474 arg1 N-glycans 1481:1489 high-mannose type N-glycans 1463:1489 high-mannose type N-glycans 1463:1489 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 3 33 theme large-scale 473:483 arg1 identification 485:498 identification 485:498 identification 485:498 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 0 34 theme Endoplasmic 0:10 arg1 degradation 42:52 Endoplasmic reticulum-associated N-glycan degradation 0:52 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 0:133 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 4 35 from N-glycans 1007:1015 arg1 mixture 1030:1036 a complex mixture 1020:1036 a complex mixture 1020:1036 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 5 36 theme c. 1073:1074 arg1 N-glycoproteins 1095:1109 c. 100 stress-related N-glycoproteins 1073:1109 c. 100 stress-related N-glycoproteins 1073:1109 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 3 37 gly glycopeptides 610:622 arg2 glycopeptides 610:622 glycopeptides 610:622 glycopeptides 610:622 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 5 38 theme reticulum 1137:1145 arg1 proteins 1161:1168 the endoplasmic reticulum (ER) resident proteins 1121:1168 the endoplasmic reticulum (ER) resident proteins 1121:1168 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 4 39 theme datasets 911:918 arg1 analysis 893:900 a rapid analysis 885:900 a rapid analysis of large datasets 885:918 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 3 40 theme data-dependent 625:638 arg1 analysis 646:653 data-dependent MS/MS analysis 625:653 data-dependent MS/MS analysis 625:653 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 4 41 theme large 905:909 arg1 datasets 911:918 large datasets 905:918 large datasets 905:918 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 0 42 theme N-glycan 33:40 arg1 degradation 42:52 Endoplasmic reticulum-associated N-glycan degradation 0:52 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 0:133 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 6 43 theme low 1333:1335 arg1 temperature 1337:1347 low temperature 1333:1347 low temperature 1333:1347 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 0 44 from stress 112:117 arg1 Arabidopsis 122:132 Arabidopsis 122:132 Arabidopsis 122:132 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 3 45 theme MS/MS 640:644 arg1 analysis 646:653 data-dependent MS/MS analysis 625:653 data-dependent MS/MS analysis 625:653 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 5 46 theme resident 1152:1159 arg1 proteins 1161:1168 the endoplasmic reticulum (ER) resident proteins 1121:1168 the endoplasmic reticulum (ER) resident proteins 1121:1168 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 4 47 theme glycoproteins 723:735 arg1 databases 698:706 In-house databases 689:706 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs 689:789 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 0 48 theme cold-upregulated 57:72 arg1 glycoproteins 74:86 cold-upregulated glycoproteins 57:86 cold-upregulated glycoproteins 57:86 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 7 49 theme N-glycans 1481:1489 arg1 degradation 1448:1458 the stress-responsive degradation 1426:1458 the stress-responsive degradation of high-mannose type N-glycans 1426:1489 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 4 50 contain containing 755:764 arg1 glycoproteins 723:735 Arabidopsis glycoproteins 711:735 Arabidopsis glycoproteins 711:735 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 50 contain containing 755:764 arg2 motifs 784:789 Asn-X-Ser/Thr/Cys motifs 766:789 Asn-X-Ser/Thr/Cys motifs 766:789 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 2 51 theme Structural 270:279 arg1 characterization 281:296 Structural characterization 270:296 Structural characterization of N-glycoproteome 270:315 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 7 52 theme chilling 1512:1519 arg1 stress 1521:1526 chilling stress 1512:1526 chilling stress 1512:1526 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 7 53 theme stress 1662:1667 arg1 conditions 1669:1678 stress conditions 1662:1678 stress conditions 1662:1678 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 3 54 theme glycoproteins 522:534 arg1 quantification 504:517 quantification 504:517 quantification 504:517 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 54 theme glycoproteins 522:534 arg1 identification 485:498 identification 485:498 identification 485:498 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 3 55 theme sensitive 441:449 arg1 method 462:467 a highly sensitive proteomics method 432:467 a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins 432:534 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 1 56 gly glycoprotein 173:184 arg1 glycoprotein 173:184 glycoprotein structure 173:194 glycoprotein structure 173:194 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 4 57 from mixture 1030:1036 arg1 quantification 957:970 quantification 957:970 quantification 957:970 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 57 from mixture 1030:1036 arg1 identification 938:951 comprehensive identification 924:951 comprehensive identification 924:951 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 3 58 gly glycoproteins 522:534 arg1 glycoproteins 522:534 glycoproteins 522:534 glycoproteins 522:534 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 2 59 theme folding 390:396 arg1 extent 372:377 the extent 368:377 the extent of protein folding and surface topology 368:417 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 0 60 theme glycoproteins 74:86 arg1 degradation 42:52 Endoplasmic reticulum-associated N-glycan degradation 0:52 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 0:133 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 4 61 theme N-glycans 1007:1015 arg1 quantification 957:970 quantification 957:970 quantification 957:970 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 61 theme N-glycans 1007:1015 arg1 identification 938:951 comprehensive identification 924:951 comprehensive identification 924:951 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 6 62 gly glycoproteins 1273:1285 arg1 glycoproteins 1273:1285 glycoproteins 1273:1285 glycoproteins 1273:1285 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 3 63 theme database 669:676 arg1 searching 678:686 automated database searching 659:686 automated database searching 659:686 We describe a highly sensitive proteomics method for large-scale identification and quantification of glycoproteins in Arabidopsis through (15) N-metabolic labeling, selective enrichment of glycopeptides, data-dependent MS/MS analysis and automated database searching. 27558752 7 64 theme type 1476:1479 arg1 N-glycans 1481:1489 high-mannose type N-glycans 1463:1489 high-mannose type N-glycans 1463:1489 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 2 65 theme protein 382:388 arg1 folding 390:396 protein folding 382:396 protein folding 382:396 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 4 66 theme complex 1022:1028 arg1 mixture 1030:1036 a complex mixture 1020:1036 a complex mixture 1020:1036 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 6 67 theme Quantitative 1204:1215 arg1 measurements 1217:1228 Quantitative measurements 1204:1228 Quantitative measurements 1204:1228 Quantitative measurements provided a molecular signature specific to glycoproteins for determining the degree of plant stress at low temperature. 27558752 1 68 theme great 157:161 arg1 impact 163:168 a great impact 155:168 a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity 155:267 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 4 69 theme size 855:858 arg1 % 822:822 20% 820:822 20% 820:822 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 69 theme size 855:858 arg1 % 830:830 90% 828:830 90% of the public database size 828:858 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 69 theme size 855:858 arg1 size 855:858 the public database size 835:858 the public database size 835:858 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 1 70 theme enzyme 253:258 arg1 activity 260:267 enzyme activity 253:267 enzyme activity 253:267 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 2 71 theme N-glycoproteome 301:315 arg1 characterization 281:296 Structural characterization 270:296 Structural characterization of N-glycoproteome 270:315 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 1 72 theme glycoprotein 173:184 arg1 structure 186:194 glycoprotein structure 173:194 glycoprotein structure 173:194 N-glycosylation has a great impact on glycoprotein structure, conformation, stability, solubility, immunogenicity and enzyme activity. 27558752 4 73 theme In-house 689:696 arg1 databases 698:706 In-house databases 689:706 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs 689:789 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 7 74 theme cold 1401:1404 arg1 treatments 1406:1415 time-course cold treatments 1389:1415 time-course cold treatments 1389:1415 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27558752 4 75 theme glycopeptides 741:753 arg1 databases 698:706 In-house databases 689:706 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs 689:789 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 0 76 gly glycoproteins 74:86 arg1 glycoproteins 74:86 cold-upregulated glycoproteins 57:86 cold-upregulated glycoproteins 57:86 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 5 77 theme stress-related 1080:1093 arg1 N-glycoproteins 1095:1109 c. 100 stress-related N-glycoproteins 1073:1109 c. 100 stress-related N-glycoproteins 1073:1109 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 4 78 theme Asn-X-Ser/Thr/Cys 766:782 arg1 motifs 784:789 Asn-X-Ser/Thr/Cys motifs 766:789 Asn-X-Ser/Thr/Cys motifs 766:789 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 4 79 theme Arabidopsis 711:721 arg1 glycoproteins 723:735 Arabidopsis glycoproteins 711:735 Arabidopsis glycoproteins 711:735 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 5 80 gly N-glycoproteins 1095:1109 arg1 N-glycoproteins 1095:1109 c. 100 stress-related N-glycoproteins 1073:1109 c. 100 stress-related N-glycoproteins 1073:1109 Proteome-wide analysis identified c. 100 stress-related N-glycoproteins, of which the endoplasmic reticulum (ER) resident proteins were examined to be up-regulated. 27558752 0 81 from degradation 42:52 arg1 response 91:98 response 91:98 response to chilling stress in Arabidopsis 91:132 Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. 27558752 4 82 theme rapid 887:891 arg1 analysis 893:900 a rapid analysis 885:900 a rapid analysis of large datasets 885:918 In-house databases of Arabidopsis glycoproteins and glycopeptides containing Asn-X-Ser/Thr/Cys motifs were constructed by reducing 20% and 90% of the public database size, respectively, to enable a rapid analysis of large datasets for comprehensive identification and quantification of glycoproteins and heterogeneous N-glycans in a complex mixture. 27558752 2 83 theme topology 410:417 arg1 extent 372:377 the extent 368:377 the extent of protein folding and surface topology 368:417 Structural characterization of N-glycoproteome has been challenging but can provide insights into the extent of protein folding and surface topology. 27558752 7 84 theme protein 1585:1591 arg1 relocation 1593:1602 protein relocation 1585:1602 protein relocation 1585:1602 Structural N-glycoproteomics following time-course cold treatments revealed the stress-responsive degradation of high-mannose type N-glycans in ER in response to chilling stress, which may aid in elucidating the cellular mechanisms of protein relocation, transport, trafficking, misfolding and degradation under stress conditions. 27614258 3 0 theme electrospray 564:575 arg1 HPAEC-MS 605:612 HPAEC-MS 605:612 HPAEC-MS 605:612 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 0 theme electrospray 564:575 arg1 spectrometry 591:602 electrospray ion trap mass spectrometry 564:602 electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 564:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 1 1 theme oligosaccharides 282:297 arg1 analysis 270:277 the analysis 266:277 the analysis of oligosaccharides without the need for prior derivatization 266:339 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 8 2 theme relative 1796:1803 arg1 quantification 1805:1818 the relative quantification 1792:1818 the relative quantification of heterogeneous samples of N-glycans 1792:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 1 3 theme amperometric 204:215 arg1 detection 217:225 pulsed amperometric detection 197:225 pulsed amperometric detection (PAD) 197:231 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 3 theme amperometric 204:215 arg1 PAD 228:230 PAD 228:230 PAD 228:230 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 3 4 theme heterogeneous 763:775 arg1 mixtures 777:784 heterogeneous mixtures 763:784 heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 763:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 4 5 theme 3mm 939:941 arg1 column 948:953 an analytical scale 3mm I.D. column 919:953 an analytical scale 3mm I.D. column 919:953 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 6 6 theme species 1370:1376 arg1 quantification 1327:1340 quantification 1327:1340 quantification 1327:1340 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 6 6 theme species 1370:1376 arg1 detection 1313:1321 detection 1313:1321 detection 1313:1321 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 3 7 theme trap 581:584 arg1 HPAEC-MS 605:612 HPAEC-MS 605:612 HPAEC-MS 605:612 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 7 theme trap 581:584 arg1 spectrometry 591:602 electrospray ion trap mass spectrometry 564:602 electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 564:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 8 8 theme samples 1837:1843 arg1 quantification 1805:1818 the relative quantification 1792:1818 the relative quantification of heterogeneous samples of N-glycans 1792:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 6 9 theme abundant 1354:1361 arg1 species 1370:1376 even low abundant glycan species 1345:1376 even low abundant glycan species 1345:1376 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 8 10 theme N-glycans 1848:1856 arg1 samples 1837:1843 heterogeneous samples 1823:1843 heterogeneous samples of N-glycans 1823:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 1 11 theme anion-exchange 148:161 arg1 chromatography 163:176 High-performance anion-exchange chromatography 131:176 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) 131:231 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 11 theme anion-exchange 148:161 arg1 method 255:260 a highly sensitive method 236:260 a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization 236:339 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 11 theme anion-exchange 148:161 arg1 HPAEC 179:183 HPAEC 179:183 HPAEC 179:183 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 4 12 theme analytical 922:931 arg1 column 948:953 an analytical scale 3mm I.D. column 919:953 an analytical scale 3mm I.D. column 919:953 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 0 13 from IgGs 125:128 arg1 N-glycans 81:89 Fc N-glycans 78:89 Fc N-glycans from heterogeneously glycosylated IgGs 78:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 0 13 from IgGs 125:128 arg1 characterization 39:54 characterization 39:54 characterization 39:54 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 0 13 from IgGs 125:128 arg1 quantification 60:73 quantification 60:73 quantification 60:73 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 3 14 theme immunoglobulin 812:825 arg1 G 827:827 immunoglobulin G 812:827 immunoglobulin G 812:827 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 6 15 theme hybrid 1428:1433 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 5 16 theme current 1142:1148 arg1 chromatograms 1150:1162 extracted ion current chromatograms 1128:1162 extracted ion current chromatograms 1128:1162 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 3 17 theme prototype 623:631 arg1 column 669:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 17 theme prototype 623:631 arg1 I.D. 648:651 1mm I.D. 644:651 1mm I.D. 644:651 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 2 18 theme reference 500:508 arg1 materials 510:518 reference materials 500:518 reference materials 500:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 3 19 theme glyco-engineered 848:863 arg1 cells 869:873 glyco-engineered CHO cells 848:873 glyco-engineered CHO cells 848:873 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 0 20 theme N-glycans 81:89 arg1 characterization 39:54 characterization 39:54 characterization 39:54 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 0 20 theme N-glycans 81:89 arg1 quantification 60:73 quantification 60:73 quantification 60:73 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 8 21 theme labelled 1903:1910 arg1 N-glycans 1912:1920 2-AB labelled N-glycans 1898:1920 2-AB labelled N-glycans 1898:1920 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 22 dep spectrometry 591:602 arg1 using 615:619 using 615:619 using a prototype mini-bore (1mm I.D.) CarboPac PA200 column 615:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 22 dep spectrometry 591:602 arg1 challenged 680:689 challenged 680:689 challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 680:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 23 theme mouse 886:890 arg1 cells 900:904 Sp2/0 mouse myeloma cells 880:904 Sp2/0 mouse myeloma cells 880:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 8 24 from performance 1749:1759 arg1 quantification 1805:1818 the relative quantification 1792:1818 the relative quantification of heterogeneous samples of N-glycans 1792:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 7 25 theme protonated 1603:1612 arg1 ions 1614:1617 protonated ions 1603:1617 protonated ions 1603:1617 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 2 26 theme peak 413:416 arg1 assignments 418:428 peak assignments 413:428 peak assignments based on the retention times of authentic standards or known peaks of reference materials 413:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 6 27 theme biantennary 1388:1398 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 5 28 theme extracted 1128:1136 arg1 chromatograms 1150:1162 extracted ion current chromatograms 1128:1162 extracted ion current chromatograms 1128:1162 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 2 29 theme authentic 462:470 arg1 standards 472:480 authentic standards 462:480 authentic standards 462:480 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 2 30 theme information 396:406 arg1 lack 379:382 the lack 375:382 the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials 375:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 5 31 theme femtomole 1204:1212 arg1 range 1214:1218 the 50-100 femtomole range 1193:1218 the 50-100 femtomole range using ion trap MS operated in positive ionization mode 1193:1273 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 0 32 gly glycosylated 112:123 arg1 IgGs 125:128 heterogeneously glycosylated IgGs 96:128 heterogeneously glycosylated IgGs 96:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 4 33 theme detection 1049:1057 arg1 limit 1040:1044 up to 8-fold improved limit 1018:1044 up to 8-fold improved limit of detection 1018:1057 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 3 34 attach derived 799:805 arg2 N-glycans 789:797 N-glycans 789:797 N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 789:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 34 attach derived 799:805 arg1 cells 900:904 Sp2/0 mouse myeloma cells 880:904 Sp2/0 mouse myeloma cells 880:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 34 attach derived 799:805 arg1 cells 869:873 glyco-engineered CHO cells 848:873 glyco-engineered CHO cells 848:873 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 34 attach derived 799:805 arg1 plasma 840:845 human plasma 834:845 human plasma 834:845 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 34 attach derived 799:805 arg1 G 827:827 immunoglobulin G 812:827 immunoglobulin G 812:827 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 7 35 theme signal-to-noise 1576:1590 arg1 ratio 1592:1596 a 65-fold improved signal-to-noise ratio 1557:1596 a 65-fold improved signal-to-noise ratio with protonated ions 1557:1617 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 2 36 theme peaks 491:495 arg1 times 453:457 the retention times 439:457 the retention times of authentic standards or known peaks of reference materials 439:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 3 37 theme CarboPac 654:661 arg1 column 669:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 37 theme CarboPac 654:661 arg1 I.D. 648:651 1mm I.D. 644:651 1mm I.D. 644:651 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 38 with on-line 551:557 arg1 HPAEC-MS 605:612 HPAEC-MS 605:612 HPAEC-MS 605:612 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 38 with on-line 551:557 arg1 spectrometry 591:602 electrospray ion trap mass spectrometry 564:602 electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 564:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 1 39 theme prior 320:324 arg1 derivatization 326:339 prior derivatization 320:339 prior derivatization 320:339 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 4 40 with performance 1001:1011 arg1 limit 1040:1044 up to 8-fold improved limit 1018:1044 up to 8-fold improved limit of detection 1018:1057 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 4 41 theme 8-fold 1024:1029 arg1 limit 1040:1044 up to 8-fold improved limit 1018:1044 up to 8-fold improved limit of detection 1018:1057 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 6 42 theme hybrid 1439:1444 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 8 43 theme quantitative 1634:1645 arg1 evaluation 1647:1656 Relative quantitative evaluation 1625:1656 Relative quantitative evaluation by single ion current chromatograms 1625:1692 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 5 44 theme positive 1250:1257 arg1 mode 1270:1273 positive ionization mode 1250:1273 positive ionization mode 1250:1273 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 2 45 with information 396:406 arg1 assignments 418:428 peak assignments 413:428 peak assignments based on the retention times of authentic standards or known peaks of reference materials 413:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 4 46 theme specific 1063:1070 arg1 N-glycans 1072:1080 specific N-glycans 1063:1080 specific N-glycans determined by PAD 1063:1098 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 8 47 theme ion 1668:1670 arg1 chromatograms 1680:1692 single ion current chromatograms 1661:1692 single ion current chromatograms 1661:1692 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 48 theme based 717:721 arg1 method 723:728 the analytical separation based method 691:728 the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 691:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 4 49 dep 8-fold 1024:1029 arg1 to 1021:1022 to 1021:1022 to 1021:1022 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 5 50 theme trap 1230:1233 arg1 MS 1235:1236 ion trap MS 1226:1236 ion trap MS operated in positive ionization mode 1226:1273 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 7 51 theme lithiated 1514:1522 arg1 adducts 1536:1542 lithiated or sodiated adducts 1514:1542 lithiated or sodiated adducts 1514:1542 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 3 52 theme analytical 695:704 arg1 method 723:728 the analytical separation based method 691:728 the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 691:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 7 53 theme N-glycans 1501:1509 arg1 detection 1488:1496 the detection 1484:1496 the detection of N-glycans as lithiated or sodiated adducts 1484:1542 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 3 54 theme structural 738:747 arg1 assignment 749:758 the structural assignment 734:758 the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 734:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 1 55 theme pulsed 197:202 arg1 detection 217:225 pulsed amperometric detection 197:225 pulsed amperometric detection (PAD) 197:231 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 55 theme pulsed 197:202 arg1 PAD 228:230 PAD 228:230 PAD 228:230 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 6 56 dep detection 1313:1321 arg1 the 1309:1311 the 1309:1311 the 1309:1311 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 6 57 theme mannose 1419:1425 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 3 58 theme mass 586:589 arg1 HPAEC-MS 605:612 HPAEC-MS 605:612 HPAEC-MS 605:612 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 58 theme mass 586:589 arg1 spectrometry 591:602 electrospray ion trap mass spectrometry 564:602 electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 564:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 59 theme mixtures 777:784 arg1 assignment 749:758 the structural assignment 734:758 the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 734:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 60 theme ion 577:579 arg1 HPAEC-MS 605:612 HPAEC-MS 605:612 HPAEC-MS 605:612 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 60 theme ion 577:579 arg1 spectrometry 591:602 electrospray ion trap mass spectrometry 564:602 electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 564:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 4 61 theme scale 933:937 arg1 column 948:953 an analytical scale 3mm I.D. column 919:953 an analytical scale 3mm I.D. column 919:953 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 6 62 theme glycan 1363:1368 arg1 species 1370:1376 even low abundant glycan species 1345:1376 even low abundant glycan species 1345:1376 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 7 63 with ratio 1592:1596 arg1 ions 1614:1617 protonated ions 1603:1617 protonated ions 1603:1617 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 4 64 theme I.D. 943:946 arg1 column 948:953 an analytical scale 3mm I.D. column 919:953 an analytical scale 3mm I.D. column 919:953 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 6 65 theme high 1414:1417 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 6 66 theme low 1350:1352 arg1 species 1370:1376 even low abundant glycan species 1345:1376 even low abundant glycan species 1345:1376 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 5 67 theme Quantitative 1101:1112 arg1 evaluation 1114:1123 Quantitative evaluation 1101:1123 Quantitative evaluation by extracted ion current chromatograms 1101:1162 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 1 68 theme High-performance 131:146 arg1 chromatography 163:176 High-performance anion-exchange chromatography 131:176 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) 131:231 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 68 theme High-performance 131:146 arg1 method 255:260 a highly sensitive method 236:260 a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization 236:339 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 68 theme High-performance 131:146 arg1 HPAEC 179:183 HPAEC 179:183 HPAEC 179:183 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 3 69 theme N-glycans 789:797 arg1 mixtures 777:784 heterogeneous mixtures 763:784 heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 763:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 70 theme mini-bore 633:641 arg1 column 669:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 70 theme mini-bore 633:641 arg1 I.D. 648:651 1mm I.D. 644:651 1mm I.D. 644:651 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 71 theme Sp2/0 880:884 arg1 cells 900:904 Sp2/0 mouse myeloma cells 880:904 Sp2/0 mouse myeloma cells 880:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 72 theme human 834:838 arg1 plasma 840:845 human plasma 834:845 human plasma 834:845 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 0 73 theme Fc 78:79 arg1 N-glycans 81:89 Fc N-glycans 78:89 Fc N-glycans from heterogeneously glycosylated IgGs 78:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 2 74 theme materials 510:518 arg1 standards 472:480 authentic standards 462:480 authentic standards 462:480 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 2 74 theme materials 510:518 arg1 peaks 491:495 known peaks 485:495 known peaks 485:495 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 4 75 theme superior 992:999 arg1 performance 1001:1011 a superior performance 990:1011 a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD 990:1098 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 3 76 theme CHO 865:867 arg1 cells 869:873 glyco-engineered CHO cells 848:873 glyco-engineered CHO cells 848:873 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 0 77 theme glycosylated 112:123 arg1 IgGs 125:128 heterogeneously glycosylated IgGs 96:128 heterogeneously glycosylated IgGs 96:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 5 78 theme detection 1173:1181 arg1 limits 1183:1188 detection limits 1173:1188 detection limits 1173:1188 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 8 79 theme 2-AB 1898:1901 arg1 N-glycans 1912:1920 2-AB labelled N-glycans 1898:1920 2-AB labelled N-glycans 1898:1920 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 80 theme myeloma 892:898 arg1 cells 900:904 Sp2/0 mouse myeloma cells 880:904 Sp2/0 mouse myeloma cells 880:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 0 81 from quantification 60:73 arg1 IgGs 125:128 heterogeneously glycosylated IgGs 96:128 heterogeneously glycosylated IgGs 96:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 6 82 theme complex-type 1400:1411 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 8 83 theme N-glycans 1912:1920 arg1 HILIC-UPLC 1884:1893 HILIC-UPLC 1884:1893 HILIC-UPLC 1884:1893 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 8 83 theme N-glycans 1912:1920 arg1 HPAEC-PAD 1870:1878 HPAEC-PAD 1870:1878 HPAEC-PAD 1870:1878 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 4 84 theme mini-bore 960:968 arg1 column 970:975 the mini-bore column 956:975 the mini-bore column 956:975 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 0 85 from characterization 39:54 arg1 IgGs 125:128 heterogeneously glycosylated IgGs 96:128 heterogeneously glycosylated IgGs 96:128 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 1 86 theme sensitive 245:253 arg1 method 255:260 a highly sensitive method 236:260 a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization 236:339 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 1 86 theme sensitive 245:253 arg1 chromatography 163:176 High-performance anion-exchange chromatography 131:176 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) 131:231 High-performance anion-exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) is a highly sensitive method for the analysis of oligosaccharides without the need for prior derivatization. 27614258 0 87 theme mini-bore 9:17 arg1 HPAEC-MS/MS 19:29 mini-bore HPAEC-MS/MS 9:29 mini-bore HPAEC-MS/MS 9:29 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 5 88 theme ion 1138:1140 arg1 chromatograms 1150:1162 extracted ion current chromatograms 1128:1162 extracted ion current chromatograms 1128:1162 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 4 89 theme improved 1031:1038 arg1 limit 1040:1044 up to 8-fold improved limit 1018:1044 up to 8-fold improved limit of detection 1018:1057 Compared to an analytical scale 3mm I.D. column, the mini-bore column demonstrated a superior performance with up to 8-fold improved limit of detection for specific N-glycans determined by PAD. 27614258 2 90 theme chemical 387:394 arg1 information 396:406 chemical information 387:406 chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials 387:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 2 91 theme retention 443:451 arg1 times 453:457 the retention times 439:457 the retention times of authentic standards or known peaks of reference materials 439:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 8 92 theme heterogeneous 1823:1835 arg1 samples 1837:1843 heterogeneous samples 1823:1843 heterogeneous samples of N-glycans 1823:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 5 93 theme ion 1226:1228 arg1 MS 1235:1236 ion trap MS 1226:1236 ion trap MS operated in positive ionization mode 1226:1273 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 2 94 theme known 485:489 arg1 peaks 491:495 known peaks 485:495 known peaks 485:495 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 8 95 theme Relative 1625:1632 arg1 evaluation 1647:1656 Relative quantitative evaluation 1625:1656 Relative quantitative evaluation by single ion current chromatograms 1625:1692 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 96 theme 1mm 644:646 arg1 column 669:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 96 theme 1mm 644:646 arg1 I.D. 648:651 1mm I.D. 644:651 1mm I.D. 644:651 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 6 97 theme bisected 1446:1453 arg1 structures 1455:1464 high mannose, hybrid and hybrid bisected structures 1414:1464 structures 1455:1464 In our hands HPAEC-MS/MS allowed the detection and quantification of even low abundant glycan species including biantennary complex-type, high mannose, hybrid and hybrid bisected structures. 27614258 7 98 theme improved 1567:1574 arg1 ratio 1592:1596 a 65-fold improved signal-to-noise ratio 1557:1596 a 65-fold improved signal-to-noise ratio with protonated ions 1557:1617 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 0 99 dep characterization 39:54 arg1 the 35:37 the 35:37 the 35:37 Applying mini-bore HPAEC-MS/MS for the characterization and quantification of Fc N-glycans from heterogeneously glycosylated IgGs. 27614258 2 100 theme standards 472:480 arg1 times 453:457 the retention times 439:457 the retention times of authentic standards or known peaks of reference materials 439:518 However, the method suffers from the lack of chemical information with peak assignments based on the retention times of authentic standards or known peaks of reference materials. 27614258 7 101 theme sodiated 1527:1534 arg1 adducts 1536:1542 lithiated or sodiated adducts 1514:1542 lithiated or sodiated adducts 1514:1542 In comparison to the detection of N-glycans as lithiated or sodiated adducts, we obtained a 65-fold improved signal-to-noise ratio with protonated ions only. 27614258 8 102 theme single 1661:1666 arg1 chromatograms 1680:1692 single ion current chromatograms 1661:1692 single ion current chromatograms 1661:1692 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 103 theme PA200 663:667 arg1 column 669:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 a prototype mini-bore (1mm I.D.) CarboPac PA200 column 621:674 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 3 103 theme PA200 663:667 arg1 I.D. 648:651 1mm I.D. 644:651 1mm I.D. 644:651 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27614258 8 104 theme excellent 1739:1747 arg1 performance 1749:1759 an excellent performance 1736:1759 an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans 1736:1856 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 5 105 theme ionization 1259:1268 arg1 mode 1270:1273 positive ionization mode 1250:1273 positive ionization mode 1250:1273 Quantitative evaluation by extracted ion current chromatograms revealed detection limits in the 50-100 femtomole range using ion trap MS operated in positive ionization mode. 27614258 8 106 theme current 1672:1678 arg1 chromatograms 1680:1692 single ion current chromatograms 1661:1692 single ion current chromatograms 1661:1692 Relative quantitative evaluation by single ion current chromatograms was successfully applied and demonstrated an excellent performance with respect to selectivity in the relative quantification of heterogeneous samples of N-glycans compared to HPAEC-PAD and HILIC-UPLC of 2-AB labelled N-glycans. 27614258 3 107 theme separation 706:715 arg1 method 723:728 the analytical separation based method 691:728 the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells 691:904 Here we applied HPAEC coupled on-line with electrospray ion trap mass spectrometry (HPAEC-MS) using a prototype mini-bore (1mm I.D.) CarboPac PA200 column and challenged the analytical separation based method for the structural assignment of heterogeneous mixtures of N-glycans derived from immunoglobulin G from human plasma, glyco-engineered CHO cells, and Sp2/0 mouse myeloma cells. 27867116 9 0 theme rapid 1668:1672 arg1 identification 1674:1687 their automatic and rapid identification 1648:1687 identification 1674:1687 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 3 1 gly glycoforms 671:680 arg1 glycopeptide 658:669 glycopeptide glycoforms 658:680 glycopeptide glycoforms with the maximum number of identification percentages 658:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 9 2 gly glycosites 1724:1733 arg2 glycosites 1724:1733 protein glycosites 1716:1733 protein glycosites 1716:1733 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 2 gly glycosites 1724:1733 arg2 glycans 1764:1770 the glycans 1760:1770 the glycans 1760:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 2 gly glycosites 1724:1733 arg1 glycans 1764:1770 the glycans 1760:1770 the glycans 1760:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 1 3 theme O-glycopeptide 261:274 arg1 detection 221:229 detection 221:229 detection 221:229 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 3 theme O-glycopeptide 261:274 arg1 identification 236:249 identification 236:249 identification 236:249 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 3 theme O-glycopeptide 261:274 arg1 separation 209:218 separation 209:218 separation 209:218 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 4 theme engine 194:199 arg1 potential 128:136 the potential 124:136 the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins 124:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 9 5 theme protein 1716:1722 arg1 glycans 1764:1770 the glycans 1760:1770 the glycans 1760:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 5 theme protein 1716:1722 arg1 glycosites 1724:1733 protein glycosites 1716:1733 protein glycosites 1716:1733 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 3 6 theme acquisition 577:587 arg1 strategies 589:598 several acquisition strategies 569:598 several acquisition strategies 569:598 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 9 7 theme glycoproteomics-based 1512:1532 arg1 methodology 1534:1544 a simple and high-throughput glycoproteomics-based methodology 1483:1544 a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans 1483:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 1 8 dep N- 254:255 arg1 glycoforms 276:285 glycoforms 276:285 glycoforms 276:285 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 7 9 theme glycopeptide 1371:1382 arg1 species 1384:1390 certain glycopeptide species 1363:1390 certain glycopeptide species 1363:1390 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 1 10 gly glycoproteins 299:311 arg1 glycoproteins 299:311 standard glycoproteins 290:311 standard glycoproteins 290:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 5 11 gly glycosylated 913:924 arg1 map 926:928 a general glycosylated map 903:928 a general glycosylated map of a given protein 903:947 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 4 12 theme proposed 741:748 arg1 simple 762:767 simple 762:767 simple 762:767 The proposed strategy is simple to set up with the technology platforms commonly used in proteomic labs. 27867116 4 12 theme proposed 741:748 arg1 strategy 750:757 The proposed strategy 737:757 The proposed strategy 737:757 The proposed strategy is simple to set up with the technology platforms commonly used in proteomic labs. 27867116 8 13 with ProteomeXchange 1416:1430 arg1 identifier 1437:1446 identifier PXD003578 1437:1456 identifier PXD003578 1437:1456 Data are available via ProteomeXchange with identifier PXD003578. 27867116 1 14 from separation 209:218 arg1 glycoproteins 299:311 standard glycoproteins 290:311 standard glycoproteins 290:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 5 15 theme given 935:939 arg1 protein 941:947 a given protein 933:947 a given protein 933:947 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 9 16 from identification 1674:1687 arg1 basis 1607:1611 the basis 1603:1611 the basis of the number of sialic acids 1603:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 17 theme glycans 1764:1770 arg1 structure 1747:1755 structure 1747:1755 structure 1747:1755 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 17 theme glycans 1764:1770 arg1 glycans 1764:1770 the glycans 1760:1770 the glycans 1760:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 17 theme glycans 1764:1770 arg1 type 1738:1741 type 1738:1741 type 1738:1741 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 17 theme glycans 1764:1770 arg1 glycosites 1724:1733 protein glycosites 1716:1733 protein glycosites 1716:1733 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 6 18 theme peptides 1205:1212 arg1 number 1195:1200 the total number 1185:1200 the total number of peptides identified 1185:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 1 19 from identification 236:249 arg1 glycoproteins 299:311 standard glycoproteins 290:311 standard glycoproteins 290:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 20 theme nano-UPLC-LTQ-FT-MS 141:159 arg1 potential 128:136 the potential 124:136 the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins 124:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 5 21 gly glycosylated 995:1006 arg1 structures 1008:1017 their corresponding glycosylated structures 975:1017 their corresponding glycosylated structures 975:1017 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 6 22 theme number 1195:1200 arg1 % 1180:1180 78% 1178:1180 78% of the total number of peptides identified 1178:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 6 22 theme number 1195:1200 arg1 number 1195:1200 the total number 1185:1200 the total number of peptides identified 1185:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 5 23 theme glycosylated 913:924 arg1 map 926:928 a general glycosylated map 903:928 a general glycosylated map of a given protein 903:947 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 0 24 theme search 89:94 arg1 engine 96:101 proteomic search engine 79:101 proteomic search engine 79:101 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 1 25 theme proteomic 177:185 arg1 engine 194:199 the Byonic™ proteomic search engine 165:199 the Byonic™ proteomic search engine 165:199 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 2 26 theme sialic 532:537 arg1 acids 539:543 sialic acids 532:543 sialic acids 532:543 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 27 gly glycoprotein 413:424 arg1 glycoprotein 413:424 the glycoprotein digest 409:431 the glycoprotein digest 409:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 5 28 theme glycosylated 995:1006 arg1 structures 1008:1017 their corresponding glycosylated structures 975:1017 their corresponding glycosylated structures 975:1017 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 1 29 gly O-glycopeptide 261:274 arg2 O-glycopeptide 261:274 O-glycopeptide 261:274 O-glycopeptide 261:274 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 2 30 theme glycoforms 466:475 arg1 separation 366:375 the separation 362:375 the separation of the glycopeptides present in the glycoprotein digest 362:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 30 theme glycoforms 466:475 arg1 baseline-resolution 439:457 a baseline-resolution 437:457 a baseline-resolution of the glycoforms of the same glycopeptide 437:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 31 gly glycopeptides 384:396 arg2 glycopeptides 384:396 the glycopeptides 380:396 the glycopeptides present in the glycoprotein digest 380:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 32 theme present 398:404 arg1 glycopeptides 384:396 the glycopeptides 380:396 the glycopeptides present in the glycoprotein digest 380:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 1 33 theme standard 290:297 arg1 glycoproteins 299:311 standard glycoproteins 290:311 standard glycoproteins 290:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 6 34 theme phase 1074:1078 arg1 approach 1094:1101 a gas phase fractionation approach 1068:1101 a gas phase fractionation approach 1068:1101 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 9 35 gly glycopeptide 1576:1587 arg2 glycopeptide 1576:1587 glycopeptide glycoforms 1576:1598 glycopeptide glycoforms 1576:1598 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 6 36 from proteins 1150:1157 arg1 peptides 1122:1129 136 unique peptides 1111:1129 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified 1111:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 0 37 theme rapid 14:18 arg1 identification 20:33 rapid identification 14:33 rapid identification 14:33 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 9 38 theme simple 1485:1490 arg1 methodology 1534:1544 a simple and high-throughput glycoproteomics-based methodology 1483:1544 a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans 1483:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 39 theme number 1620:1625 arg1 basis 1607:1611 the basis 1603:1611 the basis of the number of sialic acids 1603:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 2 40 theme glycopeptide 489:500 arg1 glycoforms 466:475 the glycoforms 462:475 the glycoforms of the same glycopeptide 462:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 3 41 theme glycoforms 671:680 arg1 characterization 638:653 characterization 638:653 characterization 638:653 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 3 41 theme glycoforms 671:680 arg1 detection 624:632 detection 624:632 detection 624:632 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 9 42 dep SIGNIFICANCE 1459:1470 arg1 propose 1475:1481 propose 1475:1481 propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans 1475:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 5 43 theme rapid 884:888 arg1 obtention 890:898 the straightforward and rapid obtention 860:898 the straightforward and rapid obtention 860:898 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 2 44 gly glycopeptide 489:500 arg2 glycopeptide 489:500 the same glycopeptide 480:500 the same glycopeptide 480:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 45 from present 398:404 arg1 digest 426:431 the glycoprotein digest 409:431 the glycoprotein digest 409:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 7 46 theme glycopeptide 1273:1284 arg1 step 1297:1300 an extra glycopeptide enrichment step 1264:1300 an extra glycopeptide enrichment step 1264:1300 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 9 47 theme sialic 1630:1635 arg1 acids 1637:1641 sialic acids 1630:1641 sialic acids 1630:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 2 48 theme nanoACQUITY 335:345 arg1 column 347:352 a BEH C18 nanoACQUITY column 325:352 a BEH C18 nanoACQUITY column 325:352 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 49 gly glycoforms 466:475 arg1 glycopeptide 489:500 the same glycopeptide 480:500 the same glycopeptide 480:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 50 from digest 426:431 arg1 present 398:404 present 398:404 present 398:404 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 1 51 dep separation 209:218 arg1 the 205:207 the 205:207 the 205:207 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 9 52 from separation 1562:1571 arg1 basis 1607:1611 the basis 1603:1611 the basis of the number of sialic acids 1603:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 3 53 gly glycopeptide 658:669 arg2 glycopeptide 658:669 glycopeptide glycoforms 658:680 glycopeptide glycoforms with the maximum number of identification percentages 658:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 3 54 theme identification 709:722 arg1 percentages 724:734 identification percentages 709:734 identification percentages 709:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 2 55 theme BEH 327:329 arg1 column 347:352 a BEH C18 nanoACQUITY column 325:352 a BEH C18 nanoACQUITY column 325:352 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 9 56 theme prior 1697:1701 arg1 knowledge 1703:1711 prior knowledge 1697:1711 prior knowledge of protein glycosites or type and structure of the glycans 1697:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 5 57 theme map 926:928 arg1 obtention 890:898 the straightforward and rapid obtention 860:898 the straightforward and rapid obtention 860:898 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 7 58 theme extra 1267:1271 arg1 step 1297:1300 an extra glycopeptide enrichment step 1264:1300 an extra glycopeptide enrichment step 1264:1300 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 1 59 theme search 187:192 arg1 engine 194:199 the Byonic™ proteomic search engine 165:199 the Byonic™ proteomic search engine 165:199 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 3 60 theme several 569:575 arg1 strategies 589:598 several acquisition strategies 569:598 several acquisition strategies 569:598 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 9 61 theme glycosites 1724:1733 arg1 knowledge 1703:1711 prior knowledge 1697:1711 prior knowledge of protein glycosites or type and structure of the glycans 1697:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 62 theme type 1738:1741 arg1 knowledge 1703:1711 prior knowledge 1697:1711 prior knowledge of protein glycosites or type and structure of the glycans 1697:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 3 63 with glycoforms 671:680 arg1 number 699:704 the maximum number 687:704 the maximum number of identification percentages 687:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 5 64 theme straightforward 864:878 arg1 obtention 890:898 the straightforward and rapid obtention 860:898 the straightforward and rapid obtention 860:898 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 7 65 gly glycopeptide 1371:1382 arg2 glycopeptide 1371:1382 certain glycopeptide species 1363:1390 certain glycopeptide species 1363:1390 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 2 66 theme acids 539:543 arg1 number 522:527 the number 518:527 the number of sialic acids 518:543 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 5 67 gly glycosites 960:969 arg2 glycosites 960:969 glycosites 960:969 glycosites 960:969 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 9 68 theme high-throughput 1496:1510 arg1 methodology 1534:1544 a simple and high-throughput glycoproteomics-based methodology 1483:1544 a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans 1483:1770 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 7 69 theme certain 1363:1369 arg1 species 1384:1390 certain glycopeptide species 1363:1390 certain glycopeptide species 1363:1390 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 5 70 theme protein 941:947 arg1 map 926:928 a general glycosylated map 903:928 a general glycosylated map of a given protein 903:947 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 9 71 theme glycopeptide 1576:1587 arg1 glycoforms 1589:1598 glycopeptide glycoforms 1576:1598 glycopeptide glycoforms 1576:1598 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 0 72 theme proteomic 79:87 arg1 engine 96:101 proteomic search engine 79:101 proteomic search engine 79:101 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 5 73 theme general 905:911 arg1 map 926:928 a general glycosylated map 903:928 a general glycosylated map of a given protein 903:947 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 6 74 theme total 1189:1193 arg1 number 1195:1200 the total number 1185:1200 the total number of peptides identified 1185:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 1 75 theme Byonic™ 169:175 arg1 engine 194:199 the Byonic™ proteomic search engine 165:199 the Byonic™ proteomic search engine 165:199 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 0 76 gly glycopeptides 38:50 arg2 glycopeptides 38:50 glycopeptides 38:50 glycopeptides 38:50 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 5 77 theme corresponding 981:993 arg1 structures 1008:1017 their corresponding glycosylated structures 975:1017 their corresponding glycosylated structures 975:1017 The method allows the straightforward and rapid obtention of a general glycosylated map of a given protein, including glycosites and their corresponding glycosylated structures. 27867116 6 78 theme MS 1024:1025 arg1 strategy 1027:1034 The MS strategy 1020:1034 The MS strategy 1020:1034 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 9 79 theme glycoforms 1589:1598 arg1 separation 1562:1571 the separation 1558:1571 the separation of glycopeptide glycoforms on the basis of the number of sialic acids 1558:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 9 79 theme glycoforms 1589:1598 arg1 identification 1674:1687 their automatic and rapid identification 1648:1687 identification 1674:1687 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 4 80 theme technology 788:797 arg1 platforms 799:807 the technology platforms 784:807 the technology platforms commonly used in proteomic labs 784:839 The proposed strategy is simple to set up with the technology platforms commonly used in proteomic labs. 27867116 6 81 theme fractionation 1080:1092 arg1 approach 1094:1101 a gas phase fractionation approach 1068:1101 a gas phase fractionation approach 1068:1101 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 9 82 gly glycoforms 1589:1598 arg1 glycopeptide 1576:1587 glycopeptide glycoforms 1576:1598 glycopeptide glycoforms 1576:1598 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 2 83 theme glycopeptides 384:396 arg1 separation 366:375 the separation 362:375 the separation of the glycopeptides present in the glycoprotein digest 362:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 83 theme glycopeptides 384:396 arg1 baseline-resolution 439:457 a baseline-resolution 437:457 a baseline-resolution of the glycoforms of the same glycopeptide 437:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 4 84 theme proteomic 826:834 arg1 labs 836:839 proteomic labs 826:839 proteomic labs 826:839 The proposed strategy is simple to set up with the technology platforms commonly used in proteomic labs. 27867116 2 85 dep number 522:527 arg1 basis 509:513 basis 509:513 basis 509:513 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 85 dep number 522:527 arg1 the 505:507 the 505:507 the 505:507 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 6 86 theme gas 1070:1072 arg1 approach 1094:1101 a gas phase fractionation approach 1068:1101 a gas phase fractionation approach 1068:1101 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 3 87 theme glycopeptide 658:669 arg1 glycoforms 671:680 glycopeptide glycoforms 658:680 glycopeptide glycoforms with the maximum number of identification percentages 658:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 0 88 theme glycopeptides 38:50 arg1 Automatic 0:8 Automatic 0:8 Automatic 0:8 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 0 88 theme glycopeptides 38:50 arg1 identification 20:33 rapid identification 14:33 rapid identification 14:33 Automatic and rapid identification of glycopeptides by nano-UPLC-LTQ-FT-MS and proteomic search engine. 27867116 3 89 dep detection 624:632 arg1 the 620:622 the 620:622 the 620:622 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 7 90 theme enrichment 1286:1295 arg1 step 1297:1300 an extra glycopeptide enrichment step 1264:1300 an extra glycopeptide enrichment step 1264:1300 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 2 91 theme same 484:487 arg1 glycopeptide 489:500 the same glycopeptide 480:500 the same glycopeptide 480:500 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 92 theme glycoprotein 413:424 arg1 digest 426:431 the glycoprotein digest 409:431 the glycoprotein digest 409:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 7 93 gly glycopeptide 1273:1284 arg2 glycopeptide 1273:1284 an extra glycopeptide enrichment step 1264:1300 an extra glycopeptide enrichment step 1264:1300 Moreover, the method does not require an extra glycopeptide enrichment step, thus preventing the bias that this step could cause towards certain glycopeptide species. 27867116 9 94 theme acids 1637:1641 arg1 number 1620:1625 the number 1616:1625 the number of sialic acids 1616:1641 SIGNIFICANCE We propose a simple and high-throughput glycoproteomics-based methodology that allows the separation of glycopeptide glycoforms on the basis of the number of sialic acids, and their automatic and rapid identification without prior knowledge of protein glycosites or type and structure of the glycans. 27867116 6 95 theme standard 1141:1148 arg1 proteins 1150:1157 four standard proteins 1136:1157 four standard proteins 1136:1157 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 3 96 theme maximum 691:697 arg1 number 699:704 the maximum number 687:704 the maximum number of identification percentages 687:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 2 97 theme column 347:352 arg1 use 318:320 The use 314:320 The use of a BEH C18 nanoACQUITY column 314:352 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 1 98 from detection 221:229 arg1 glycoproteins 299:311 standard glycoproteins 290:311 standard glycoproteins 290:311 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 2 99 theme C18 331:333 arg1 column 347:352 a BEH C18 nanoACQUITY column 325:352 a BEH C18 nanoACQUITY column 325:352 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 100 attach present 398:404 arg2 glycopeptides 384:396 the glycopeptides 380:396 the glycopeptides present in the glycoprotein digest 380:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 2 100 attach present 398:404 arg1 digest 426:431 the glycoprotein digest 409:431 the glycoprotein digest 409:431 The use of a BEH C18 nanoACQUITY column allowed the separation of the glycopeptides present in the glycoprotein digest and a baseline-resolution of the glycoforms of the same glycopeptide on the basis of the number of sialic acids. 27867116 6 101 theme unique 1115:1120 arg1 peptides 1122:1129 136 unique peptides 1111:1129 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified 1111:1223 The MS strategy selected in this work, based on a gas phase fractionation approach, led to 136 unique peptides from four standard proteins, which represented 78% of the total number of peptides identified. 27867116 3 102 theme percentages 724:734 arg1 number 699:704 the maximum number 687:704 the maximum number of identification percentages 687:734 Moreover, we evaluated several acquisition strategies in order to improve the detection and characterization of glycopeptide glycoforms with the maximum number of identification percentages. 27867116 1 103 theme N- 254:255 arg1 detection 221:229 detection 221:229 detection 221:229 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 103 theme N- 254:255 arg1 identification 236:249 identification 236:249 identification 236:249 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 27867116 1 103 theme N- 254:255 arg1 separation 209:218 separation 209:218 separation 209:218 Here we demonstrate the potential of nano-UPLC-LTQ-FT-MS and the Byonic™ proteomic search engine for the separation, detection, and identification of N- and O-glycopeptide glycoforms in standard glycoproteins. 25164466 4 0 theme O-linked 736:743 arg1 N-acetylglucosamine 745:763 O-linked N-acetylglucosamine 736:763 O-linked N-acetylglucosamine (O-GlcNAc) 736:774 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 0 theme O-linked 736:743 arg1 O-GlcNAc 766:773 O-GlcNAc 766:773 O-GlcNAc 766:773 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 1 gly glycosylation 158:170 arg1 components 188:197 its components 184:197 its components 184:197 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 1 gly glycosylation 158:170 arg1 membrane 141:148 the plasma membrane 130:148 the plasma membrane 130:148 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 2 theme membrane 141:148 arg1 organization 114:125 the spatial organization 102:125 the spatial organization of the plasma membrane 102:148 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 2 theme membrane 141:148 arg1 patterns 172:179 the glycosylation patterns 154:179 the glycosylation patterns of its components 154:197 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 4 3 theme homogeneous 640:650 arg1 distribution 652:663 a homogeneous distribution 638:663 a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface 638:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 4 with labeling 382:389 arg1 precursors 459:468 modified monosaccharide precursors 435:468 modified monosaccharide precursors 435:468 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 2 4 with labeling 382:389 arg1 fluorophores 482:493 organic fluorophores 474:493 organic fluorophores 474:493 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 5 from densities 839:847 arg1 surface 889:895 each cell surface 879:895 each cell surface 879:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 6 theme N-acetylglucosamine 745:763 arg1 distribution 652:663 a homogeneous distribution 638:663 a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface 638:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 7 used used 346:349 arg2 strategy 333:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 3 8 theme single-molecule 585:599 arg1 sensitivity 601:611 single-molecule sensitivity 585:611 single-molecule sensitivity 585:611 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 4 9 from distribution 652:663 arg1 lines 828:832 different cell lines 813:832 different cell lines with densities of several million glycans on each cell surface 813:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 10 theme cell 823:826 arg1 lines 828:832 different cell lines 813:832 different cell lines with densities of several million glycans on each cell surface 813:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 11 theme plasma 785:790 arg1 proteins 801:808 -modified plasma membrane proteins 775:808 -modified plasma membrane proteins 775:808 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 12 theme -modified 775:783 arg1 proteins 801:808 -modified plasma membrane proteins 775:808 -modified plasma membrane proteins 775:808 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 13 theme glycoconjugates 414:428 arg1 labeling 382:389 the efficient and specific labeling 355:389 the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores 355:493 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 3 14 with glycans 572:578 arg1 sensitivity 601:611 single-molecule sensitivity 585:611 single-molecule sensitivity 585:611 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 4 15 theme N-acetylmannosamine 668:686 arg1 ManNAc 689:694 N-acetylmannosamine (ManNAc)- 668:696 N-acetylmannosamine (ManNAc)- 668:696 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 16 theme glycosylation 158:170 arg1 patterns 172:179 the glycosylation patterns 154:179 the glycosylation patterns of its components 154:197 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 3 17 theme membrane 563:570 arg1 glycans 572:578 plasma membrane glycans 556:578 plasma membrane glycans with single-molecule sensitivity 556:611 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 4 18 theme GalNAc 722:727 arg1 distribution 652:663 a homogeneous distribution 638:663 a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface 638:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 19 theme cells 79:83 arg1 physiology 65:74 the physiology 61:74 the physiology of cells 61:83 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 0 20 theme Super-resolution 0:15 arg1 imaging 17:23 Super-resolution imaging 0:23 Super-resolution imaging of plasma membrane glycans 0:50 Super-resolution imaging of plasma membrane glycans. 25164466 4 21 theme membrane 792:799 arg1 proteins 801:808 -modified plasma membrane proteins 775:808 -modified plasma membrane proteins 775:808 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 22 theme efficient 359:367 arg1 labeling 382:389 the efficient and specific labeling 355:389 the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores 355:493 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 0 23 theme plasma 28:33 arg1 glycans 44:50 plasma membrane glycans 28:50 plasma membrane glycans 28:50 Super-resolution imaging of plasma membrane glycans. 25164466 4 24 link O-linked 736:743 arg1 N-acetylglucosamine 745:763 O-linked N-acetylglucosamine 736:763 O-linked N-acetylglucosamine (O-GlcNAc) 736:774 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 24 link O-linked 736:743 arg1 O-GlcNAc 766:773 O-GlcNAc 766:773 O-GlcNAc 766:773 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 25 theme modified 435:442 arg1 precursors 459:468 modified monosaccharide precursors 435:468 modified monosaccharide precursors 435:468 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 26 with lines 828:832 arg1 densities 839:847 densities 839:847 densities of several million glycans on each cell surface 839:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 27 theme components 188:197 arg1 organization 114:125 the spatial organization 102:125 the spatial organization of the plasma membrane 102:148 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 27 theme components 188:197 arg1 patterns 172:179 the glycosylation patterns 154:179 the glycosylation patterns of its components 154:197 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 0 28 theme glycans 44:50 arg1 imaging 17:23 Super-resolution imaging 0:23 Super-resolution imaging of plasma membrane glycans 0:50 Super-resolution imaging of plasma membrane glycans. 25164466 1 29 theme components 267:276 arg1 size 236:239 size 236:239 size 236:239 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 29 theme components 267:276 arg1 composition 246:256 composition 246:256 composition 246:256 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 1 29 theme components 267:276 arg1 distribution 222:233 distribution 222:233 distribution 222:233 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 2 30 theme membrane-associated 394:412 arg1 glycoconjugates 414:428 membrane-associated glycoconjugates 394:428 membrane-associated glycoconjugates 394:428 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 0 31 theme membrane 35:42 arg1 glycans 44:50 plasma membrane glycans 28:50 plasma membrane glycans 28:50 Super-resolution imaging of plasma membrane glycans. 25164466 2 32 theme monosaccharide 444:457 arg1 precursors 459:468 modified monosaccharide precursors 435:468 modified monosaccharide precursors 435:468 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 33 from surface 889:895 arg1 densities 839:847 densities 839:847 densities of several million glycans on each cell surface 839:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 34 theme N-acetylgalactosamine 699:719 arg1 GalNAc 722:727 N-acetylgalactosamine (GalNAc)- 699:729 N-acetylgalactosamine (GalNAc)- 699:729 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 35 dep studying 209:216 arg1 controlled 88:97 controlled 88:97 is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components 85:197 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 3 36 used used 538:541 arg2 imaging 526:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 3 37 theme Super-resolution 496:511 arg1 imaging 526:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 2 38 theme organic 474:480 arg1 fluorophores 482:493 organic fluorophores 474:493 organic fluorophores 474:493 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 39 theme cell 884:887 arg1 surface 889:895 each cell surface 879:895 each cell surface 879:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 40 theme spatial 106:112 arg1 organization 114:125 the spatial organization 102:125 the spatial organization of the plasma membrane 102:148 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 4 41 dep ManNAc 689:694 arg1 proteins 801:808 -modified plasma membrane proteins 775:808 -modified plasma membrane proteins 775:808 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 42 theme several 852:858 arg1 glycans 868:874 several million glycans 852:874 several million glycans on each cell surface 852:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 43 theme chemical 315:322 arg1 strategy 333:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 44 theme different 813:821 arg1 lines 828:832 different cell lines 813:832 different cell lines with densities of several million glycans on each cell surface 813:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 4 45 from glycans 868:874 arg1 surface 889:895 each cell surface 879:895 each cell surface 879:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 2 46 theme bioorthogonal 301:313 arg1 strategy 333:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 3 47 theme fluorescence 513:524 arg1 imaging 526:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging 496:532 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 2 48 theme reporter 324:331 arg1 strategy 333:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy 299:340 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 49 theme glycans 868:874 arg1 densities 839:847 densities 839:847 densities of several million glycans on each cell surface 839:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 3 50 theme plasma 556:561 arg1 glycans 572:578 plasma membrane glycans 556:578 plasma membrane glycans with single-molecule sensitivity 556:611 Super-resolution fluorescence imaging was used to visualize plasma membrane glycans with single-molecule sensitivity. 25164466 2 51 theme specific 373:380 arg1 labeling 382:389 the efficient and specific labeling 355:389 the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores 355:493 A bioorthogonal chemical reporter strategy was used for the efficient and specific labeling of membrane-associated glycoconjugates with modified monosaccharide precursors and organic fluorophores. 25164466 4 52 theme million 860:866 arg1 glycans 868:874 several million glycans 852:874 several million glycans on each cell surface 852:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 25164466 1 53 theme plasma 134:139 arg1 membrane 141:148 the plasma membrane 130:148 the plasma membrane 130:148 Much of the physiology of cells is controlled by the spatial organization of the plasma membrane and the glycosylation patterns of its components, however, studying the distribution, size, and composition of these components remains challenging. 25164466 4 54 theme ManNAc 689:694 arg1 distribution 652:663 a homogeneous distribution 638:663 a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface 638:895 Our results demonstrate a homogeneous distribution of N-acetylmannosamine (ManNAc)-, N-acetylgalactosamine (GalNAc)-, and O-linked N-acetylglucosamine (O-GlcNAc)-modified plasma membrane proteins in different cell lines with densities of several million glycans on each cell surface. 27997162 9 0 theme potential 1862:1870 arg1 application 1689:1699 This novel application 1678:1699 This novel application of MGE 1678:1706 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 9 0 theme potential 1862:1870 arg1 drug 1872:1875 a potential drug 1860:1875 a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo 1860:1967 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 8 1 from levels 1544:1549 arg1 NCAM 1554:1557 NCAM 1554:1557 NCAM 1554:1557 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 8 1 from levels 1544:1549 arg1 mice 1568:1571 adult mice 1562:1571 adult mice 1562:1571 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 6 2 from administration 1064:1077 arg1 mice 1099:1102 mice 1099:1102 mice (C57BL/6J and BALB/cByJ) 1099:1127 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 2 from administration 1064:1077 arg1 BALB/cByJ 1118:1126 BALB/cByJ 1118:1126 BALB/cByJ 1118:1126 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 2 from administration 1064:1077 arg1 C57BL/6J 1105:1112 C57BL/6J 1105:1112 C57BL/6J 1105:1112 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 8 3 theme hybrid 1645:1650 arg1 administration 1605:1618 administration 1605:1618 administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut 1605:1675 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 6 4 theme nonhybrid 1258:1266 arg1 Ac4ManNAz 1277:1285 the nonhybrid molecule Ac4ManNAz 1254:1285 the nonhybrid molecule Ac4ManNAz 1254:1285 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 2 5 theme CNS 348:350 arg1 injury 352:357 CNS injury 348:357 CNS injury 348:357 Dysregulation in biosynthesis of sialo-glycoconjugates is known to be associated with neurological disorders, CNS injury, and brain cancer. 27997162 7 6 theme polysialic 1468:1477 arg1 polySia 1485:1491 polySia 1485:1491 polySia 1485:1491 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 7 6 theme polysialic 1468:1477 arg1 acid 1479:1482 polysialic acid 1468:1482 polysialic acid (polySia) 1468:1492 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 6 7 theme N-azidoacetyl-neuraminic 1162:1185 arg1 glycoproteins 1209:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 4 8 theme CNH 649:651 arg1 molecules 654:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 5 9 theme neuroactive 871:881 arg1 acid 926:929 valproic acid 917:929 valproic acid 917:929 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 9 theme neuroactive 871:881 arg1 acid 911:914 nicotinic acid 901:914 nicotinic acid 901:914 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 9 theme neuroactive 871:881 arg1 carriers 883:890 neuroactive carriers 871:890 neuroactive carriers 871:890 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 9 theme neuroactive 871:881 arg1 acid 954:957 theophylline-7-acetic acid 932:957 theophylline-7-acetic acid 932:957 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 9 theme neuroactive 871:881 arg1 choline 964:970 choline 964:970 choline 964:970 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 9 10 theme neural 1900:1905 arg1 biosynthesis 1914:1925 neural glycan biosynthesis 1900:1925 neural glycan biosynthesis 1900:1925 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 9 11 theme biosynthesis 1914:1925 arg1 modulation 1886:1895 modulation 1886:1895 modulation of neural glycan biosynthesis and thus function 1886:1943 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 3 12 theme sialoglycans 533:544 arg1 investigations 515:528 direct investigations 508:528 direct investigations of sialoglycans in brain 508:553 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 9 13 theme MGE 1704:1706 arg1 application 1689:1699 This novel application 1678:1699 This novel application of MGE 1678:1706 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 9 13 theme MGE 1704:1706 arg1 drug 1872:1875 a potential drug 1860:1875 a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo 1860:1967 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 4 14 theme hybrid 641:646 arg1 molecules 654:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 1 15 theme brain 197:201 arg1 development 203:213 brain development 197:213 brain development 197:213 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 5 16 theme valproic 917:924 arg1 carriers 883:890 neuroactive carriers 871:890 neuroactive carriers 871:890 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 16 theme valproic 917:924 arg1 acid 926:929 valproic acid 917:929 valproic acid 917:929 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 7 17 theme neuroactive 1349:1359 arg1 carriers 1361:1368 Successful neuroactive carriers 1338:1368 Successful neuroactive carriers 1338:1368 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 6 18 theme glycoproteins 1209:1221 arg1 expression 1148:1157 robust expression 1141:1157 robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart 1141:1245 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 19 theme NeuAz 1294:1298 arg1 expression 1300:1309 NeuAz expression 1294:1309 NeuAz expression 1294:1309 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 3 20 theme direct 508:513 arg1 investigations 515:528 direct investigations 508:528 direct investigations of sialoglycans in brain 508:553 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 1 21 theme central 146:152 arg1 CNS 170:172 CNS 170:172 CNS 170:172 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 1 21 theme central 146:152 arg1 system 162:167 the central nervous system 142:167 the central nervous system (CNS) 142:173 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 4 22 theme carrier-mediated 679:694 arg1 systems 706:712 carrier-mediated transport systems 679:712 carrier-mediated transport systems available at the blood-brain barrier 679:749 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 0 23 theme Nervous 89:95 arg1 System 97:102 the Central Nervous System 77:102 the Central Nervous System 77:102 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 3 24 from investigations 515:528 arg1 brain 549:553 brain 549:553 brain 549:553 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 6 25 theme acid 1187:1190 arg1 glycoproteins 1209:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 1 26 from abundant 130:137 arg1 CNS 170:172 CNS 170:172 CNS 170:172 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 1 26 from abundant 130:137 arg1 system 162:167 the central nervous system 142:167 the central nervous system (CNS) 142:173 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 4 27 theme blood-brain 731:741 arg1 barrier 743:749 the blood-brain barrier 727:749 the blood-brain barrier 727:749 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 2 28 theme brain 364:368 arg1 cancer 370:375 brain cancer 364:375 brain cancer 364:375 Dysregulation in biosynthesis of sialo-glycoconjugates is known to be associated with neurological disorders, CNS injury, and brain cancer. 27997162 0 29 theme Carbohydrate-Neuroactive 0:23 arg1 Strategy 32:39 Carbohydrate-Neuroactive Hybrid Strategy 0:39 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo 0:110 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 6 30 theme Intravenous 1052:1062 arg1 administration 1064:1077 Intravenous administration 1052:1077 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) 1052:1127 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 1 31 theme Sialic 113:118 arg1 acids 120:124 Sialic acids 113:124 Sialic acids 113:124 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 2 32 from Dysregulation 238:250 arg1 biosynthesis 255:266 biosynthesis 255:266 biosynthesis of sialo-glycoconjugates 255:291 Dysregulation in biosynthesis of sialo-glycoconjugates is known to be associated with neurological disorders, CNS injury, and brain cancer. 27997162 9 33 theme brain 1813:1817 arg1 applications 1827:1838 brain mapping applications 1813:1838 brain mapping applications 1813:1838 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 0 34 theme Metabolic 45:53 arg1 Engineering 62:72 Metabolic Glycan Engineering 45:72 Metabolic Glycan Engineering of the Central Nervous System in Vivo 45:110 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 5 35 theme human 1016:1020 arg1 cells 1037:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 3 36 theme Metabolic 378:386 arg1 MGE 408:410 MGE 408:410 MGE 408:410 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 3 36 theme Metabolic 378:386 arg1 engineering 395:405 Metabolic glycan engineering 378:405 Metabolic glycan engineering (MGE) 378:411 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 3 37 theme roles 473:477 arg1 study 453:457 study 453:457 study of biological roles of glycans 453:488 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 7 38 theme cell 1504:1507 arg1 NCAM 1529:1532 NCAM 1529:1532 NCAM 1529:1532 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 7 38 theme cell 1504:1507 arg1 molecules 1518:1526 neural cell adhesion molecules 1497:1526 neural cell adhesion molecules (NCAM) 1497:1533 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 8 39 theme adult 1562:1566 arg1 mice 1568:1571 adult mice 1562:1571 adult mice 1562:1571 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 4 40 theme vein 777:780 arg1 injection 782:790 tail vein injection 772:790 tail vein injection in mice 772:798 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 5 41 theme SH-SY5Y 1007:1013 arg1 cells 1037:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 3 42 theme glycans 482:488 arg1 roles 473:477 biological roles 462:477 biological roles of glycans 462:488 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 7 43 theme acid 1479:1482 arg1 modulation 1454:1463 modulation 1454:1463 modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM) 1454:1533 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 6 44 theme molecules 1086:1094 arg1 administration 1064:1077 Intravenous administration 1052:1077 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) 1052:1127 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 45 theme molecule 1268:1275 arg1 Ac4ManNAz 1277:1285 the nonhybrid molecule Ac4ManNAz 1254:1285 the nonhybrid molecule Ac4ManNAz 1254:1285 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 2 46 theme sialo-glycoconjugates 271:291 arg1 biosynthesis 255:266 biosynthesis 255:266 biosynthesis of sialo-glycoconjugates 255:291 Dysregulation in biosynthesis of sialo-glycoconjugates is known to be associated with neurological disorders, CNS injury, and brain cancer. 27997162 5 47 theme nicotinic 901:909 arg1 acid 911:914 nicotinic acid 901:914 nicotinic acid 901:914 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 47 theme nicotinic 901:909 arg1 carriers 883:890 neuroactive carriers 871:890 neuroactive carriers 871:890 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 8 48 theme Ac3ManNBut-nicotinate 1623:1643 arg1 hybrid 1645:1650 Ac3ManNBut-nicotinate hybrid 1623:1650 Ac3ManNBut-nicotinate hybrid 1623:1650 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 7 49 from modulation 1454:1463 arg1 NCAM 1529:1532 NCAM 1529:1532 NCAM 1529:1532 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 7 49 from modulation 1454:1463 arg1 molecules 1518:1526 neural cell adhesion molecules 1497:1526 neural cell adhesion molecules (NCAM) 1497:1533 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 9 50 theme novel 1683:1687 arg1 application 1689:1699 This novel application 1678:1699 This novel application of MGE 1678:1706 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 9 50 theme novel 1683:1687 arg1 drug 1872:1875 a potential drug 1860:1875 a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo 1860:1967 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 6 51 theme robust 1141:1146 arg1 expression 1148:1157 robust expression 1141:1157 robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart 1141:1245 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 9 52 theme glycan 1907:1912 arg1 biosynthesis 1914:1925 neural glycan biosynthesis 1900:1925 neural glycan biosynthesis 1900:1925 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 9 53 theme function 1936:1943 arg1 modulation 1886:1895 modulation 1886:1895 modulation of neural glycan biosynthesis and thus function 1886:1943 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 4 54 theme carbohydrate-neuroactive 616:639 arg1 molecules 654:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 carbohydrate-neuroactive hybrid (CNH) molecules 616:662 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 4 55 theme transport 696:704 arg1 systems 706:712 carrier-mediated transport systems 679:712 carrier-mediated transport systems available at the blood-brain barrier 679:749 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 9 56 theme noninvasive 1726:1736 arg1 tool 1738:1741 a noninvasive tool 1724:1741 a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications 1724:1838 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 6 57 theme -carrying 1199:1207 arg1 glycoproteins 1209:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 4 58 theme available 714:722 arg1 systems 706:712 carrier-mediated transport systems 679:712 carrier-mediated transport systems available at the blood-brain barrier 679:749 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 0 59 theme Central 81:87 arg1 System 97:102 the Central Nervous System 77:102 the Central Nervous System 77:102 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 4 60 from injection 782:790 arg1 mice 795:798 mice 795:798 mice 795:798 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 7 61 theme Successful 1338:1347 arg1 carriers 1361:1368 Successful neuroactive carriers 1338:1368 Successful neuroactive carriers 1338:1368 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 0 62 theme System 97:102 arg1 Engineering 62:72 Metabolic Glycan Engineering 45:72 Metabolic Glycan Engineering of the Central Nervous System in Vivo 45:110 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 1 63 theme nervous 154:160 arg1 CNS 170:172 CNS 170:172 CNS 170:172 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 1 63 theme nervous 154:160 arg1 system 162:167 the central nervous system 142:167 the central nervous system (CNS) 142:173 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 9 64 theme brain 1761:1765 arg1 glycosylation 1767:1779 brain glycosylation 1761:1779 brain glycosylation 1761:1779 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 1 65 from system 162:167 arg1 abundant 130:137 abundant 130:137 abundant 130:137 Sialic acids are abundant in the central nervous system (CNS) and are essential for brain development, learning, and memory. 27997162 8 66 with administration 1605:1618 arg1 Ac4ManNBut 1666:1675 Ac4ManNBut 1666:1675 Ac4ManNBut 1666:1675 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 6 67 theme CNH 1082:1084 arg1 molecules 1086:1094 CNH molecules 1082:1094 CNH molecules 1082:1094 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 3 68 theme bioorthogonal 417:429 arg1 ligation 431:438 bioorthogonal ligation 417:438 bioorthogonal ligation 417:438 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 0 69 theme Hybrid 25:30 arg1 Strategy 32:39 Carbohydrate-Neuroactive Hybrid Strategy 0:39 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo 0:110 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 4 70 from barrier 743:749 arg1 available 714:722 available 714:722 available 714:722 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 5 71 theme theophylline-7-acetic 932:952 arg1 carriers 883:890 neuroactive carriers 871:890 neuroactive carriers 871:890 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 71 theme theophylline-7-acetic 932:952 arg1 acid 954:957 theophylline-7-acetic acid 932:957 theophylline-7-acetic acid 932:957 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 8 72 theme PolySia 1536:1542 arg1 levels 1544:1549 PolySia levels 1536:1549 PolySia levels on NCAM in adult mice 1536:1571 PolySia levels on NCAM in adult mice were reduced significantly upon administration of Ac3ManNBut-nicotinate hybrid, but not with Ac4ManNBut. 27997162 5 73 theme Peracetylated 801:813 arg1 Ac4ManNAz 844:852 Ac4ManNAz 844:852 Ac4ManNAz 844:852 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 5 73 theme Peracetylated 801:813 arg1 N-azidoacetyl-d-mannosamine 815:841 Peracetylated N-azidoacetyl-d-mannosamine 801:841 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, 801:971 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 6 74 from expression 1148:1157 arg1 heart 1241:1245 heart 1241:1245 heart 1241:1245 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 74 from expression 1148:1157 arg1 brain 1231:1235 brain 1231:1235 brain 1231:1235 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 0 75 theme Glycan 55:60 arg1 Engineering 62:72 Metabolic Glycan Engineering 45:72 Metabolic Glycan Engineering of the Central Nervous System in Vivo 45:110 Carbohydrate-Neuroactive Hybrid Strategy for Metabolic Glycan Engineering of the Central Nervous System in Vivo. 27997162 6 76 theme NeuAz 1193:1197 arg1 glycoproteins 1209:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 9 77 theme mapping 1819:1825 arg1 applications 1827:1838 brain mapping applications 1813:1838 brain mapping applications 1813:1838 This novel application of MGE not only offers a noninvasive tool for investigating brain glycosylation, which could be developed in to brain mapping applications, but also serves as a potential drug by which modulation of neural glycan biosynthesis and thus function can be achieved in vivo. 27997162 5 78 theme neuroblastoma 1022:1034 arg1 cells 1037:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 SH-SY5Y (human neuroblastoma) cells 1007:1041 Peracetylated N-azidoacetyl-d-mannosamine (Ac4ManNAz) conjugated with neuroactive carriers, namely, nicotinic acid, valproic acid, theophylline-7-acetic acid, and choline, were synthesized and evaluated in SH-SY5Y (human neuroblastoma) cells for MGE. 27997162 3 79 theme glycan 388:393 arg1 MGE 408:410 MGE 408:410 MGE 408:410 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 3 79 theme glycan 388:393 arg1 engineering 395:405 Metabolic glycan engineering 378:405 Metabolic glycan engineering (MGE) 378:411 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 6 80 dep mice 1099:1102 arg1 mice 1099:1102 mice 1099:1102 mice (C57BL/6J and BALB/cByJ) 1099:1127 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 80 dep mice 1099:1102 arg1 BALB/cByJ 1118:1126 BALB/cByJ 1118:1126 BALB/cByJ 1118:1126 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 6 80 dep mice 1099:1102 arg1 C57BL/6J 1105:1112 C57BL/6J 1105:1112 C57BL/6J 1105:1112 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27997162 2 81 theme neurological 324:335 arg1 disorders 337:345 neurological disorders 324:345 neurological disorders 324:345 Dysregulation in biosynthesis of sialo-glycoconjugates is known to be associated with neurological disorders, CNS injury, and brain cancer. 27997162 7 82 theme adhesion 1509:1516 arg1 NCAM 1529:1532 NCAM 1529:1532 NCAM 1529:1532 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 7 82 theme adhesion 1509:1516 arg1 molecules 1518:1526 neural cell adhesion molecules 1497:1526 neural cell adhesion molecules (NCAM) 1497:1533 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 4 83 theme tail 772:775 arg1 injection 782:790 tail vein injection 772:790 tail vein injection in mice 772:798 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 3 84 theme biological 462:471 arg1 roles 473:477 biological roles 462:477 biological roles of glycans 462:488 Metabolic glycan engineering (MGE) and bioorthogonal ligation have enabled study of biological roles of glycans in vivo; however, direct investigations of sialoglycans in brain have been intractable. 27997162 7 85 theme neural 1497:1502 arg1 NCAM 1529:1532 NCAM 1529:1532 NCAM 1529:1532 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 7 85 theme neural 1497:1502 arg1 molecules 1518:1526 neural cell adhesion molecules 1497:1526 neural cell adhesion molecules (NCAM) 1497:1533 Successful neuroactive carriers were then conjugated with N-butanoyl-d-mannosamine (ManNBut) with a goal to achieve modulation of polysialic acid (polySia) on neural cell adhesion molecules (NCAM). 27997162 4 86 theme simple 590:595 arg1 strategy 597:604 a simple strategy 588:604 a simple strategy 588:604 We report a simple strategy utilizing carbohydrate-neuroactive hybrid (CNH) molecules, which exploit carrier-mediated transport systems available at the blood-brain barrier, to access brain via tail vein injection in mice. 27997162 6 87 gly glycoproteins 1209:1221 arg1 glycoproteins 1209:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins 1162:1221 Intravenous administration of CNH molecules in mice (C57BL/6J and BALB/cByJ) resulted in robust expression of N-azidoacetyl-neuraminic acid (NeuAz)-carrying glycoproteins in both brain and heart, while the nonhybrid molecule Ac4ManNAz showed NeuAz expression in heart but not in brain. 27851982 7 0 theme key 1614:1616 arg1 factor 1618:1623 the key factor 1610:1623 the key factor modulating the functioning of CDH 1610:1657 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 7 0 theme key 1614:1616 arg1 repulsion 1597:1605 the interdomain electrostatic repulsion 1567:1605 the interdomain electrostatic repulsion 1567:1605 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 6 1 theme electron 1404:1411 arg1 state 1439:1443 its inactive (open) and interdomain electron transfer-capable (closed) state 1368:1443 state 1439:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 2 2 theme polysaccharide 388:401 arg1 monooxygenase 403:415 lytic polysaccharide monooxygenase 382:415 lytic polysaccharide monooxygenase 382:415 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 8 3 theme repulsion 1770:1778 arg1 role 1755:1758 the role 1751:1758 the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells 1751:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 3 4 with combination 691:701 arg1 rhizopuspepsin 725:738 rhizopuspepsin 725:738 rhizopuspepsin 725:738 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 7 5 theme electrostatic 1583:1595 arg1 factor 1618:1623 the key factor 1610:1623 the key factor modulating the functioning of CDH 1610:1657 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 7 5 theme electrostatic 1583:1595 arg1 repulsion 1597:1605 the interdomain electrostatic repulsion 1567:1605 the interdomain electrostatic repulsion 1567:1605 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 3 6 theme mass 749:752 arg1 spectrometry 754:765 native mass spectrometry 742:765 native mass spectrometry with ion mobility and electrostatics calculations 742:815 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 7 theme structural 560:569 arg1 aspects 571:577 the structural aspects 556:577 the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin) 556:739 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 1 8 theme lytic 192:196 arg1 monooxygenase 213:225 lytic polysaccharide monooxygenase 192:225 lytic polysaccharide monooxygenase 192:225 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 3 9 theme electrostatics 789:802 arg1 calculations 804:815 electrostatics calculations 789:815 electrostatics calculations 789:815 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 5 10 theme Electrostatics 949:962 arg1 calculations 964:975 Electrostatics calculations 949:975 Electrostatics calculations 949:975 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 3 11 theme domain 594:599 arg1 interaction 601:611 the domain interaction 590:611 the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin) 590:739 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 8 12 theme biotechnological 1886:1901 arg1 potential 1903:1911 their biotechnological potential 1880:1911 their biotechnological potential in biosensors and biofuel cells 1880:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 2 13 theme Interdomain 272:282 arg1 transfer 293:300 Interdomain electron transfer 272:300 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, 272:416 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 1 14 theme cellulose 249:257 arg1 degradation 259:269 cellulose degradation 249:269 cellulose degradation 249:269 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 6 15 theme CDH 1356:1358 arg1 Transition 1342:1351 CONCLUSIONS Transition 1330:1351 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state 1330:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 8 16 from role 1755:1758 arg1 relevant 1856:1863 relevant 1856:1863 relevant 1856:1863 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 8 16 from role 1755:1758 arg1 transfer 1808:1815 the interdomain electron transfer 1783:1815 the interdomain electron transfer 1783:1815 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 6 17 from interface 1532:1540 arg1 changes 1472:1478 changes 1472:1478 changes in the protein surface electrostatics at the domain interface 1472:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 6 17 from interface 1532:1540 arg1 electrostatics 1503:1516 the protein surface electrostatics 1483:1516 the protein surface electrostatics at the domain interface 1483:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 4 18 from changes 865:871 arg1 accessibility 884:896 solvent accessibility 876:896 solvent accessibility 876:896 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 4 18 from changes 865:871 arg1 bonding 911:917 hydrogen bonding 902:917 hydrogen bonding 902:917 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 5 19 theme charge 1021:1026 arg1 neutralization 1028:1041 charge neutralization 1021:1041 charge neutralization by protonation 1021:1056 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 6 20 theme surface 1495:1501 arg1 electrostatics 1503:1516 the protein surface electrostatics 1483:1516 the protein surface electrostatics at the domain interface 1483:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 5 21 theme linker 1225:1230 arg1 region 1232:1237 the linker region 1221:1237 the linker region 1221:1237 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 1 22 theme extracellular 151:163 arg1 dehydrogenase 119:131 BACKGROUND Cellobiose dehydrogenase 97:131 BACKGROUND Cellobiose dehydrogenase (CDH) 97:137 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 22 theme extracellular 151:163 arg1 oxidoreductase 165:178 a fungal extracellular oxidoreductase 142:178 a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation 142:269 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 0 23 from transfer 21:28 arg1 dehydrogenase 44:56 cellobiose dehydrogenase 33:56 cellobiose dehydrogenase 33:56 Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics. 27851982 5 24 theme pH. 1162:1164 arg1 addition 1169:1176 neutral pH. In addition 1154:1176 neutral pH. In addition 1154:1176 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 8 25 from transfer 1808:1815 arg1 dehydrogenases 1831:1844 cellobiose dehydrogenases 1820:1844 cellobiose dehydrogenases 1820:1844 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 5 26 theme extensive 1192:1200 arg1 O-glycosylation 1202:1216 extensive O-glycosylation 1192:1216 extensive O-glycosylation in the linker region 1192:1237 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 4 27 theme RESULTS 828:834 arg1 HDX-MS 836:841 RESULTS HDX-MS 828:841 RESULTS HDX-MS 828:841 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 0 28 theme Interdomain 0:10 arg1 transfer 21:28 Interdomain electron transfer 0:28 Interdomain electron transfer in cellobiose dehydrogenase 0:56 Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics. 27851982 8 29 dep SIGNIFICANCE 1668:1679 arg1 provide 1717:1723 provide 1717:1723 provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells 1717:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 1 30 theme Cellobiose 108:117 arg1 oxidoreductase 165:178 a fungal extracellular oxidoreductase 142:178 a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation 142:269 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 30 theme Cellobiose 108:117 arg1 CDH 134:136 CDH 134:136 CDH 134:136 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 30 theme Cellobiose 108:117 arg1 dehydrogenase 119:131 BACKGROUND Cellobiose dehydrogenase 97:131 BACKGROUND Cellobiose dehydrogenase (CDH) 97:137 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 5 31 from O-glycosylation 1202:1216 arg1 region 1232:1237 the linker region 1221:1237 the linker region 1221:1237 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 32 theme papain-mediated 1295:1309 arg1 separation 1318:1327 papain-mediated domain separation 1295:1327 papain-mediated domain separation 1295:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 4 33 theme solvent 876:882 arg1 accessibility 884:896 solvent accessibility 876:896 solvent accessibility 876:896 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 0 34 theme cellobiose 33:42 arg1 dehydrogenase 44:56 cellobiose dehydrogenase 33:56 cellobiose dehydrogenase 33:56 Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics. 27851982 6 35 theme domain 1525:1530 arg1 interface 1532:1540 the domain interface 1521:1540 the domain interface 1521:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 3 36 theme proteolytic 672:682 arg1 setup 684:688 improved proteolytic setup 663:688 improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin) 663:739 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 2 37 theme lytic 382:386 arg1 monooxygenase 403:415 lytic polysaccharide monooxygenase 382:415 lytic polysaccharide monooxygenase 382:415 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 5 38 theme exact 1271:1275 arg1 point 1286:1290 the long-unknown exact cleavage point 1254:1290 the long-unknown exact cleavage point in papain-mediated domain separation 1254:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 3 39 with spectrometry 754:765 arg1 mobility 776:783 ion mobility 772:783 ion mobility 772:783 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 39 with spectrometry 754:765 arg1 calculations 804:815 electrostatics calculations 789:815 electrostatics calculations 789:815 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 8 40 theme experimental 1725:1736 arg1 evidence 1738:1745 experimental evidence 1725:1745 experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells 1725:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 8 41 theme interdomain 1787:1797 arg1 relevant 1856:1863 relevant 1856:1863 relevant 1856:1863 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 8 41 theme interdomain 1787:1797 arg1 transfer 1808:1815 the interdomain electron transfer 1783:1815 the interdomain electron transfer 1783:1815 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 3 42 dep setup 684:688 arg1 combination 691:701 combination 691:701 combination of nepenthesin-1 with rhizopuspepsin 691:738 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 5 43 theme electrostatic 1107:1119 arg1 repulsion 1121:1129 higher electrostatic repulsion 1100:1129 higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation 1100:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 3 44 theme native 742:747 arg1 spectrometry 754:765 native mass spectrometry 742:765 native mass spectrometry with ion mobility and electrostatics calculations 742:815 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 6 45 theme transfer-capable 1413:1428 arg1 state 1439:1443 its inactive (open) and interdomain electron transfer-capable (closed) state 1368:1443 state 1439:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 4 46 theme interdomain 926:936 arg1 interface 938:946 the interdomain interface 922:946 the interdomain interface 922:946 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 6 47 theme interdomain 1392:1402 arg1 state 1439:1443 its inactive (open) and interdomain electron transfer-capable (closed) state 1368:1443 state 1439:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 8 48 theme cellobiose 1820:1829 arg1 dehydrogenases 1831:1844 cellobiose dehydrogenases 1820:1844 cellobiose dehydrogenases 1820:1844 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 7 49 theme interdomain 1571:1581 arg1 factor 1618:1623 the key factor 1610:1623 the key factor modulating the functioning of CDH 1610:1657 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 7 49 theme interdomain 1571:1581 arg1 repulsion 1597:1605 the interdomain electrostatic repulsion 1567:1605 the interdomain electrostatic repulsion 1567:1605 Our study confirms that the interdomain electrostatic repulsion is the key factor modulating the functioning of CDH. 27851982 8 50 theme charge 1763:1768 arg1 repulsion 1770:1778 charge repulsion 1763:1778 charge repulsion 1763:1778 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 3 51 theme ion 772:774 arg1 mobility 776:783 ion mobility 772:783 ion mobility 772:783 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 2 52 theme cytochrome 325:334 arg1 domain 336:341 the flavin and cytochrome domain 310:341 domain 336:341 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 4 53 theme hydrogen 902:909 arg1 bonding 911:917 hydrogen bonding 902:917 hydrogen bonding 902:917 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 27851982 5 54 dep domains 1143:1149 arg1 identified 1243:1252 identified 1243:1252 identified the long-unknown exact cleavage point in papain-mediated domain separation 1243:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 54 dep domains 1143:1149 arg1 uncovered 1182:1190 uncovered 1182:1190 uncovered extensive O-glycosylation in the linker region 1182:1237 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 55 from point 1286:1290 arg1 separation 1318:1327 papain-mediated domain separation 1295:1327 papain-mediated domain separation 1295:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 1 56 theme polysaccharide 198:211 arg1 monooxygenase 213:225 lytic polysaccharide monooxygenase 192:225 lytic polysaccharide monooxygenase 192:225 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 5 57 gly O-glycosylation 1202:1216 arg1 region 1232:1237 the linker region 1221:1237 the linker region 1221:1237 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 57 gly O-glycosylation 1202:1216 arg1 addition 1169:1176 neutral pH. In addition 1154:1176 neutral pH. In addition 1154:1176 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 8 58 from potential 1903:1911 arg1 biosensors 1916:1925 biosensors 1916:1925 biosensors 1916:1925 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 8 58 from potential 1903:1911 arg1 cells 1939:1943 biofuel cells 1931:1943 biofuel cells 1931:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 3 59 theme hydrogen/deuterium 621:638 arg1 HDX-MS 650:655 HDX-MS 650:655 HDX-MS 650:655 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 59 theme hydrogen/deuterium 621:638 arg1 CDH 616:618 CDH 616:618 CDH 616:618 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 59 theme hydrogen/deuterium 621:638 arg1 exchange 640:647 hydrogen/deuterium exchange 621:647 hydrogen/deuterium exchange (HDX-MS) 621:656 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 60 used used 822:825 arg2 spectrometry 754:765 native mass spectrometry 742:765 native mass spectrometry with ion mobility and electrostatics calculations 742:815 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 6 61 theme closed 1431:1436 arg1 state 1439:1443 its inactive (open) and interdomain electron transfer-capable (closed) state 1368:1443 state 1439:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 5 62 theme neutral 1154:1160 arg1 addition 1169:1176 neutral pH. In addition 1154:1176 neutral pH. In addition 1154:1176 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 2 63 theme flavin 314:319 arg1 domain 336:341 the flavin and cytochrome domain 310:341 domain 336:341 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 6 64 theme CONCLUSIONS 1330:1340 arg1 Transition 1342:1351 CONCLUSIONS Transition 1330:1351 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state 1330:1443 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 3 65 from interaction 601:611 arg1 CDH 616:618 CDH 616:618 CDH 616:618 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 3 65 from interaction 601:611 arg1 exchange 640:647 hydrogen/deuterium exchange 621:647 hydrogen/deuterium exchange (HDX-MS) 621:656 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 0 66 theme surface 73:79 arg1 electrostatics 81:94 surface electrostatics 73:94 surface electrostatics 73:94 Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics. 27851982 5 67 theme In 1166:1167 arg1 addition 1169:1176 neutral pH. In addition 1154:1176 neutral pH. In addition 1154:1176 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 8 68 theme biofuel 1931:1937 arg1 cells 1939:1943 biofuel cells 1931:1943 biofuel cells 1931:1943 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 1 69 theme fungal 144:149 arg1 dehydrogenase 119:131 BACKGROUND Cellobiose dehydrogenase 97:131 BACKGROUND Cellobiose dehydrogenase (CDH) 97:137 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 69 theme fungal 144:149 arg1 oxidoreductase 165:178 a fungal extracellular oxidoreductase 142:178 a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation 142:269 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 2 70 from domain 336:341 arg1 CDH 346:348 CDH 346:348 CDH 346:348 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 6 71 theme protein 1487:1493 arg1 electrostatics 1503:1516 the protein surface electrostatics 1483:1516 the protein surface electrostatics at the domain interface 1483:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 5 72 theme ion 1076:1078 arg1 mobility 1080:1087 ion mobility 1076:1087 ion mobility 1076:1087 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 8 73 theme GENERAL 1660:1666 arg1 SIGNIFICANCE 1668:1679 GENERAL SIGNIFICANCE 1660:1679 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 1660:1944 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 5 74 dep result 1009:1014 arg1 pointed 1089:1095 pointed 1089:1095 pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation 1089:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 0 75 theme electron 12:19 arg1 transfer 21:28 Interdomain electron transfer 0:28 Interdomain electron transfer in cellobiose dehydrogenase 0:56 Interdomain electron transfer in cellobiose dehydrogenase is governed by surface electrostatics. 27851982 6 76 from changes 1472:1478 arg1 interface 1532:1540 the domain interface 1521:1540 the domain interface 1521:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 6 76 from changes 1472:1478 arg1 electrostatics 1503:1516 the protein surface electrostatics 1483:1516 the protein surface electrostatics at the domain interface 1483:1540 CONCLUSIONS Transition of CDH between its inactive (open) and interdomain electron transfer-capable (closed) state is shown to be governed by changes in the protein surface electrostatics at the domain interface. 27851982 1 77 theme BACKGROUND 97:106 arg1 oxidoreductase 165:178 a fungal extracellular oxidoreductase 142:178 a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation 142:269 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 77 theme BACKGROUND 97:106 arg1 CDH 134:136 CDH 134:136 CDH 134:136 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 1 77 theme BACKGROUND 97:106 arg1 dehydrogenase 119:131 BACKGROUND Cellobiose dehydrogenase 97:131 BACKGROUND Cellobiose dehydrogenase (CDH) 97:137 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 3 78 with interaction 601:611 arg1 setup 684:688 improved proteolytic setup 663:688 improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin) 663:739 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 2 79 theme electron 284:291 arg1 transfer 293:300 Interdomain electron transfer 272:300 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, 272:416 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 2 80 theme exact 455:459 arg1 mechanism 461:469 the exact mechanism 451:469 the exact mechanism of this regulation 451:488 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 5 81 dep identified 977:986 arg1 result 1009:1014 result 1009:1014 to result from charge neutralization by protonation 1006:1056 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 81 dep identified 977:986 arg1 mobility 1080:1087 ion mobility 1076:1087 ion mobility 1076:1087 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 82 theme domain 1311:1316 arg1 separation 1318:1327 papain-mediated domain separation 1295:1327 papain-mediated domain separation 1295:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 3 83 theme improved 663:670 arg1 setup 684:688 improved proteolytic setup 663:688 improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin) 663:739 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 5 84 theme long-unknown 1258:1269 arg1 point 1286:1290 the long-unknown exact cleavage point 1254:1290 the long-unknown exact cleavage point in papain-mediated domain separation 1254:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 5 85 theme CDH 1139:1141 arg1 domains 1143:1149 CDH domains 1139:1149 CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation 1139:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 2 86 theme regulation 479:488 arg1 mechanism 461:469 the exact mechanism 451:469 the exact mechanism of this regulation 451:488 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 3 87 theme nepenthesin-1 706:718 arg1 combination 691:701 combination 691:701 combination of nepenthesin-1 with rhizopuspepsin 691:738 METHODS To investigate the structural aspects underlying the domain interaction in CDH, hydrogen/deuterium exchange (HDX-MS) with improved proteolytic setup (combination of nepenthesin-1 with rhizopuspepsin), native mass spectrometry with ion mobility and electrostatics calculations were used. 27851982 5 88 theme cleavage 1277:1284 arg1 point 1286:1290 the long-unknown exact cleavage point 1254:1290 the long-unknown exact cleavage point in papain-mediated domain separation 1254:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 1 89 with fuels 186:190 arg1 electrons 232:240 electrons 232:240 electrons 232:240 BACKGROUND Cellobiose dehydrogenase (CDH) is a fungal extracellular oxidoreductase which fuels lytic polysaccharide monooxygenase with electrons during cellulose degradation. 27851982 2 90 theme electron 365:372 arg1 flow 374:377 the electron flow 361:377 the electron flow to lytic polysaccharide monooxygenase 361:415 Interdomain electron transfer between the flavin and cytochrome domain in CDH, preceding the electron flow to lytic polysaccharide monooxygenase, is known to be pH dependent, but the exact mechanism of this regulation has not been experimentally proven so far. 27851982 8 91 theme electron 1799:1806 arg1 relevant 1856:1863 relevant 1856:1863 relevant 1856:1863 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 8 91 theme electron 1799:1806 arg1 transfer 1808:1815 the interdomain electron transfer 1783:1815 the interdomain electron transfer 1783:1815 GENERAL SIGNIFICANCE The results presented in this paper provide experimental evidence for the role of charge repulsion in the interdomain electron transfer in cellobiose dehydrogenases, which is relevant for exploiting their biotechnological potential in biosensors and biofuel cells. 27851982 5 92 theme higher 1100:1105 arg1 repulsion 1121:1129 higher electrostatic repulsion 1100:1129 higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation 1100:1327 Electrostatics calculations identified these differences to result from charge neutralization by protonation and together with ion mobility pointed at higher electrostatic repulsion between CDH domains at neutral pH. In addition, we uncovered extensive O-glycosylation in the linker region and identified the long-unknown exact cleavage point in papain-mediated domain separation. 27851982 4 93 theme pH-dependent 852:863 arg1 changes 865:871 pH-dependent changes 852:871 pH-dependent changes in solvent accessibility and hydrogen bonding 852:917 RESULTS HDX-MS revealed pH-dependent changes in solvent accessibility and hydrogen bonding at the interdomain interface. 26321095 0 0 theme clinical 82:89 arg1 outcome 91:97 clinical outcome 82:97 clinical outcome 82:97 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 0 1 from analysis 15:22 arg1 cancer 34:39 breast cancer 27:39 breast cancer 27:39 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 7 2 theme basal-like 1181:1190 arg1 subgroup 1192:1199 the basal-like subgroup 1177:1199 the basal-like subgroup 1177:1199 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 1 3 theme cancer 160:165 arg1 development 167:177 cancer development 160:177 cancer development 160:177 Glycosylation and related processes play important roles in cancer development and progression, including metastasis. 26321095 2 4 contain have 260:263 arg2 value 286:290 potential diagnostic value 265:290 potential diagnostic value 265:290 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 4 contain have 260:263 arg1 N-glycans 250:258 N-glycans 250:258 N-glycans 250:258 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 4 contain have 260:263 arg2 biomarkers 308:317 cancer serum biomarkers 295:317 cancer serum biomarkers 295:317 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 5 5 theme trigalactosylated 793:809 arg1 glycans 825:831 tri antennary trigalactosylated trisialylated glycans 779:831 tri antennary trigalactosylated trisialylated glycans 779:831 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 5 6 theme trisialylated 811:823 arg1 glycans 825:831 tri antennary trigalactosylated trisialylated glycans 779:831 tri antennary trigalactosylated trisialylated glycans 779:831 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 5 7 theme integrin-mediated 933:949 arg1 adhesion 956:963 integrin-mediated cell adhesion 933:963 integrin-mediated cell adhesion 933:963 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 5 8 theme cell 951:954 arg1 adhesion 956:963 integrin-mediated cell adhesion 933:963 integrin-mediated cell adhesion 933:963 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 6 9 theme ER 1042:1043 arg1 tumours 1054:1060 ER negative tumours 1042:1060 ER negative tumours 1042:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 3 10 theme particular 374:383 arg1 structures 400:409 particular serum N-glycan structures 374:409 particular serum N-glycan structures as important features of breast tumour biology 374:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 11 dep glycome 480:486 arg1 mRNA 514:517 mRNA 514:517 mRNA 514:517 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 11 dep glycome 480:486 arg1 the 470:472 the 470:472 the 470:472 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 11 dep glycome 480:486 arg1 miRNA 523:527 miRNA 523:527 miRNA 523:527 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 12 theme important 414:422 arg1 features 424:431 important features 414:431 important features of breast tumour biology 414:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 13 theme patients 551:558 arg1 glycome 480:486 serum glycome 474:486 serum glycome 474:486 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 13 theme patients 551:558 arg1 transcriptome 499:511 tumour transcriptome 492:511 tumour transcriptome 492:511 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 14 theme serum 385:389 arg1 structures 400:409 particular serum N-glycan structures 374:409 particular serum N-glycan structures as important features of breast tumour biology 374:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 2 15 theme serum 302:306 arg1 biomarkers 308:317 cancer serum biomarkers 295:317 cancer serum biomarkers 295:317 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 15 theme serum 302:306 arg1 value 286:290 potential diagnostic value 265:290 potential diagnostic value 265:290 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 4 16 theme data 601:604 arg1 types 582:586 these types 576:586 these types of molecular data 576:604 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 3 17 theme abundance 361:369 arg1 significance 341:352 the significance 337:352 the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology 337:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 18 theme breast 537:542 arg1 patients 551:558 104 breast cancer patients 533:558 104 breast cancer patients 533:558 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 4 19 theme functional 693:702 arg1 pathways 732:739 metabolic pathways 722:739 metabolic pathways 722:739 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 4 19 theme functional 693:702 arg1 pathways 704:711 functional pathways 693:711 functional pathways 693:711 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 4 19 theme functional 693:702 arg1 response 755:762 DNA damage response 744:762 DNA damage response 744:762 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 5 20 theme transcripts 890:900 arg1 levels 873:878 lower levels 867:878 lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion 867:963 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 8 21 theme cells 1237:1241 arg1 presence 1206:1213 The presence 1202:1213 The presence of circulating tumour cells 1202:1241 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 4 22 theme molecular 591:599 arg1 data 601:604 molecular data 591:604 molecular data 591:604 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 3 23 theme cancer 544:549 arg1 patients 551:558 104 breast cancer patients 533:558 104 breast cancer patients 533:558 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 8 24 theme glycome 1303:1309 arg1 trisialylated 1383:1395 trisialylated 1383:1395 trisialylated 1383:1395 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 24 theme glycome 1303:1309 arg1 trigalactosylated 1356:1372 trigalactosylated 1356:1372 trigalactosylated 1356:1372 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 24 theme glycome 1303:1309 arg1 di- 1375:1377 di- 1375:1377 di- 1375:1377 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 24 theme glycome 1303:1309 arg1 structures 1311:1320 several serum glycome structures 1289:1320 several serum glycome structures like bi and triantennary 1289:1345 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 24 theme glycome 1303:1309 arg1 di- 1348:1350 di- 1348:1350 di- 1348:1350 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 0 25 theme N-glycan 6:13 arg1 analysis 15:22 Serum N-glycan analysis 0:22 Serum N-glycan analysis in breast cancer 0:39 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 7 26 theme increased 1141:1149 arg1 survival 1151:1158 increased survival 1141:1158 increased survival 1141:1158 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 3 27 theme biology 450:456 arg1 features 424:431 important features 414:431 important features of breast tumour biology 414:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 0 28 theme Serum 0:4 arg1 analysis 15:22 Serum N-glycan analysis 0:22 Serum N-glycan analysis in breast cancer 0:39 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 4 29 theme damage 748:753 arg1 response 755:762 DNA damage response 744:762 DNA damage response 744:762 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 6 30 theme negative 1045:1052 arg1 tumours 1054:1060 ER negative tumours 1042:1060 ER negative tumours 1042:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 5 31 theme focal 914:918 arg1 adhesion 920:927 focal adhesion 914:927 focal adhesion 914:927 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 3 32 theme N-glycan 391:398 arg1 structures 400:409 particular serum N-glycan structures 374:409 particular serum N-glycan structures as important features of breast tumour biology 374:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 7 33 from survival 1151:1158 arg1 subgroup 1192:1199 the basal-like subgroup 1177:1199 the basal-like subgroup 1177:1199 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 6 34 theme poor 1010:1013 arg1 prognosis 1015:1023 poor prognosis 1010:1023 poor prognosis in patients with ER negative tumours 1010:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 5 35 theme lower 867:871 arg1 levels 873:878 lower levels 867:878 lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion 867:963 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 3 36 theme structures 400:409 arg1 abundance 361:369 the abundance 357:369 the abundance of particular serum N-glycan structures as important features of breast tumour biology 357:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 4 37 theme types 582:586 arg1 Integration 561:571 Integration 561:571 Integration of these types of molecular data 561:604 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 2 38 theme cancer 295:300 arg1 biomarkers 308:317 cancer serum biomarkers 295:317 cancer serum biomarkers 295:317 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 38 theme cancer 295:300 arg1 value 286:290 potential diagnostic value 265:290 potential diagnostic value 265:290 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 3 39 theme tumour 492:497 arg1 transcriptome 499:511 tumour transcriptome 492:511 tumour transcriptome 492:511 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 3 40 theme breast 436:441 arg1 biology 450:456 breast tumour biology 436:456 breast tumour biology 436:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 9 41 theme miRNA 1422:1426 arg1 levels 1439:1444 tumour miRNA expression levels 1415:1444 tumour miRNA expression levels 1415:1444 The link between tumour miRNA expression levels and N-glycan production is also examined. 26321095 7 42 theme structures 1109:1118 arg1 abundance 1068:1076 High abundance 1063:1076 High abundance of simple monoantennary glycan structures 1063:1118 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 3 43 theme serum 474:478 arg1 glycome 480:486 serum glycome 474:486 serum glycome 474:486 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 0 44 theme breast 27:32 arg1 cancer 34:39 breast cancer 27:39 breast cancer 27:39 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 5 45 gly trisialylated 811:823 arg1 glycans 825:831 tri antennary trigalactosylated trisialylated glycans 779:831 tri antennary trigalactosylated trisialylated glycans 779:831 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 3 46 theme tumour 443:448 arg1 biology 450:456 breast tumour biology 436:456 breast tumour biology 436:456 We have explored the significance of the abundance of particular serum N-glycan structures as important features of breast tumour biology by studying the serum glycome and tumour transcriptome (mRNA and miRNA) of 104 breast cancer patients. 26321095 6 47 attach linked 1000:1005 arg2 structures 979:988 These glycan structures 966:988 These glycan structures 966:988 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 6 47 attach linked 1000:1005 arg1 prognosis 1015:1023 poor prognosis 1010:1023 poor prognosis in patients with ER negative tumours 1010:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 7 48 theme High 1063:1066 arg1 abundance 1068:1076 High abundance 1063:1076 High abundance of simple monoantennary glycan structures 1063:1118 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 4 49 theme serum 650:654 arg1 glycans 656:662 serum glycans 650:662 serum glycans 650:662 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 8 50 theme serum 1297:1301 arg1 trisialylated 1383:1395 trisialylated 1383:1395 trisialylated 1383:1395 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 50 theme serum 1297:1301 arg1 trigalactosylated 1356:1372 trigalactosylated 1356:1372 trigalactosylated 1356:1372 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 50 theme serum 1297:1301 arg1 di- 1375:1377 di- 1375:1377 di- 1375:1377 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 50 theme serum 1297:1301 arg1 structures 1311:1320 several serum glycome structures 1289:1320 several serum glycome structures like bi and triantennary 1289:1345 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 50 theme serum 1297:1301 arg1 di- 1348:1350 di- 1348:1350 di- 1348:1350 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 51 theme tumour 1230:1235 arg1 cells 1237:1241 circulating tumour cells 1218:1241 circulating tumour cells 1218:1241 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 7 52 theme monoantennary 1088:1100 arg1 structures 1109:1118 simple monoantennary glycan structures 1081:1118 simple monoantennary glycan structures 1081:1118 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 1 53 theme related 118:124 arg1 processes 126:134 related processes 118:134 related processes 118:134 Glycosylation and related processes play important roles in cancer development and progression, including metastasis. 26321095 6 54 from prognosis 1015:1023 arg1 patients 1028:1035 patients 1028:1035 patients with ER negative tumours 1028:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 7 55 theme glycan 1102:1107 arg1 structures 1109:1118 simple monoantennary glycan structures 1081:1118 simple monoantennary glycan structures 1081:1118 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 4 56 theme metabolic 722:730 arg1 pathways 732:739 metabolic pathways 722:739 metabolic pathways 722:739 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 9 57 theme N-glycan 1450:1457 arg1 production 1459:1468 N-glycan production 1450:1468 N-glycan production 1450:1468 The link between tumour miRNA expression levels and N-glycan production is also examined. 26321095 2 58 theme Several 218:224 arg1 studies 226:232 Several studies 218:232 Several studies 218:232 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 9 59 theme tumour 1415:1420 arg1 levels 1439:1444 tumour miRNA expression levels 1415:1444 tumour miRNA expression levels 1415:1444 The link between tumour miRNA expression levels and N-glycan production is also examined. 26321095 4 60 theme DNA 744:746 arg1 damage 748:753 DNA damage 744:753 DNA damage response 744:762 Integration of these types of molecular data allows us to study the relationship between serum glycans and transcripts representing functional pathways, such as metabolic pathways or DNA damage response. 26321095 7 61 theme simple 1081:1086 arg1 structures 1109:1118 simple monoantennary glycan structures 1081:1118 simple monoantennary glycan structures 1081:1118 High abundance of simple monoantennary glycan structures were associated with increased survival, particularly in the basal-like subgroup. 26321095 8 62 theme several 1289:1295 arg1 trisialylated 1383:1395 trisialylated 1383:1395 trisialylated 1383:1395 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 62 theme several 1289:1295 arg1 trigalactosylated 1356:1372 trigalactosylated 1356:1372 trigalactosylated 1356:1372 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 62 theme several 1289:1295 arg1 di- 1375:1377 di- 1375:1377 di- 1375:1377 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 62 theme several 1289:1295 arg1 structures 1311:1320 several serum glycome structures 1289:1320 several serum glycome structures like bi and triantennary 1289:1345 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 8 62 theme several 1289:1295 arg1 di- 1348:1350 di- 1348:1350 di- 1348:1350 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 26321095 2 63 theme diagnostic 275:284 arg1 biomarkers 308:317 cancer serum biomarkers 295:317 cancer serum biomarkers 295:317 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 63 theme diagnostic 275:284 arg1 value 286:290 potential diagnostic value 265:290 potential diagnostic value 265:290 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 1 64 theme important 141:149 arg1 roles 151:155 important roles 141:155 important roles 141:155 Glycosylation and related processes play important roles in cancer development and progression, including metastasis. 26321095 9 65 theme expression 1428:1437 arg1 levels 1439:1444 tumour miRNA expression levels 1415:1444 tumour miRNA expression levels 1415:1444 The link between tumour miRNA expression levels and N-glycan production is also examined. 26321095 5 66 theme tumour 883:888 arg1 transcripts 890:900 tumour transcripts 883:900 tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion 883:963 We identified tri antennary trigalactosylated trisialylated glycans in serum as being associated with lower levels of tumour transcripts involved in focal adhesion and integrin-mediated cell adhesion. 26321095 2 67 theme potential 265:273 arg1 biomarkers 308:317 cancer serum biomarkers 295:317 cancer serum biomarkers 295:317 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 2 67 theme potential 265:273 arg1 value 286:290 potential diagnostic value 265:290 potential diagnostic value 265:290 Several studies have shown that N-glycans have potential diagnostic value as cancer serum biomarkers. 26321095 6 68 with patients 1028:1035 arg1 tumours 1054:1060 ER negative tumours 1042:1060 ER negative tumours 1042:1060 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 0 69 theme tumour 63:68 arg1 biology 70:76 tumour biology 63:76 tumour biology 63:76 Serum N-glycan analysis in breast cancer patients--Relation to tumour biology and clinical outcome. 26321095 6 70 theme glycan 972:977 arg1 structures 979:988 These glycan structures 966:988 These glycan structures 966:988 These glycan structures were also linked to poor prognosis in patients with ER negative tumours. 26321095 8 71 theme circulating 1218:1228 arg1 cells 1237:1241 circulating tumour cells 1218:1241 circulating tumour cells 1218:1241 The presence of circulating tumour cells was found to be significantly associated with several serum glycome structures like bi and triantennary, di- and trigalactosylated, di- and trisialylated. 27131291 0 0 theme hiemalis 88:95 arg1 endo-β-N-acetylglucosaminidase 97:126 Mucor hiemalis endo-β-N-acetylglucosaminidase 82:126 Mucor hiemalis endo-β-N-acetylglucosaminidase 82:126 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 2 1 theme linkage 458:464 arg1 structure 431:439 the structure 427:439 the structure of the glycoside linkage formed by the Endo-M transglycosylation 427:504 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 1 2 theme Mucor 277:281 arg1 Endo-M 324:329 Endo-M 324:329 Endo-M 324:329 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 2 theme Mucor 277:281 arg1 endo-β-N-acetylglucosaminidase 292:321 Mucor hiemalis endo-β-N-acetylglucosaminidase 277:321 Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 277:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 0 3 theme Mucor 82:86 arg1 endo-β-N-acetylglucosaminidase 97:126 Mucor hiemalis endo-β-N-acetylglucosaminidase 82:126 Mucor hiemalis endo-β-N-acetylglucosaminidase 82:126 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 2 4 theme C-NMR 355:359 arg1 HMBC 378:381 (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY 351:399 HMBC 378:381 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 1 5 theme hiemalis 283:290 arg1 Endo-M 324:329 Endo-M 324:329 Endo-M 324:329 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 5 theme hiemalis 283:290 arg1 endo-β-N-acetylglucosaminidase 292:321 Mucor hiemalis endo-β-N-acetylglucosaminidase 277:321 Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 277:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 2 6 dep connection 517:526 arg1 i.e. 507:510 i.e. 507:510 i.e. 507:510 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 2 7 from GlcNAc 547:552 arg1 oligosaccharide 557:571 oligosaccharide 1 557:573 oligosaccharide 1 557:573 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 2 8 theme Endo-M 480:485 arg1 transglycosylation 487:504 the Endo-M transglycosylation 476:504 the Endo-M transglycosylation 476:504 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 2 9 from GlcNAc 536:541 arg1 oligosaccharide 557:571 oligosaccharide 1 557:573 oligosaccharide 1 557:573 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 1 10 theme spectral 196:203 arg1 assignment 205:214 the complete nuclear magnetic resonance (NMR) spectral assignment 150:214 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 0 11 theme NMR 9:11 arg1 assignment 13:22 Complete NMR assignment 0:22 Complete NMR assignment of a bisecting hybrid-type oligosaccharide 0:65 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 2 12 theme glycoside 448:456 arg1 linkage 458:464 the glycoside linkage 444:464 the glycoside linkage formed by the Endo-M transglycosylation 444:504 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 2 13 theme H- 344:345 arg1 experiments 401:411 (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments 341:411 (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments 341:411 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 0 14 theme Complete 0:7 arg1 assignment 13:22 Complete NMR assignment 0:22 Complete NMR assignment of a bisecting hybrid-type oligosaccharide 0:65 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 1 15 theme bisecting 221:229 arg1 oligosaccharide 243:257 a bisecting hybrid-type oligosaccharide 1 219:259 a bisecting hybrid-type oligosaccharide 1 219:259 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 0 16 theme bisecting 29:37 arg1 oligosaccharide 51:65 a bisecting hybrid-type oligosaccharide 27:65 a bisecting hybrid-type oligosaccharide 27:65 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 1 17 theme hybrid-type 231:241 arg1 oligosaccharide 243:257 a bisecting hybrid-type oligosaccharide 1 219:259 a bisecting hybrid-type oligosaccharide 1 219:259 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 18 theme oligosaccharide 243:257 arg1 assignment 205:214 the complete nuclear magnetic resonance (NMR) spectral assignment 150:214 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 0 19 theme oligosaccharide 51:65 arg1 assignment 13:22 Complete NMR assignment 0:22 Complete NMR assignment of a bisecting hybrid-type oligosaccharide 0:65 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 0 20 theme hybrid-type 39:49 arg1 oligosaccharide 51:65 a bisecting hybrid-type oligosaccharide 27:65 a bisecting hybrid-type oligosaccharide 27:65 Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase. 27131291 1 21 theme complete 154:161 arg1 resonance 180:188 complete nuclear magnetic resonance 154:188 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 21 theme complete 154:161 arg1 NMR 191:193 NMR 191:193 NMR 191:193 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 22 theme nuclear 163:169 arg1 resonance 180:188 complete nuclear magnetic resonance 154:188 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 22 theme nuclear 163:169 arg1 NMR 191:193 NMR 191:193 NMR 191:193 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 2 23 dep H- 344:345 arg1 1 342:342 1 342:342 1 342:342 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 1 24 theme magnetic 171:178 arg1 resonance 180:188 complete nuclear magnetic resonance 154:188 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 1 24 theme magnetic 171:178 arg1 NMR 191:193 NMR 191:193 NMR 191:193 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 27131291 2 25 theme HMBC 378:381 arg1 experiments 401:411 (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments 341:411 (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments 341:411 Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. 27131291 1 26 theme resonance 180:188 arg1 assignment 205:214 the complete nuclear magnetic resonance (NMR) spectral assignment 150:214 the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M) 150:330 This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). 25048706 9 0 theme unique 2007:2012 arg1 structures 2014:2023 27 unique structures 2004:2023 27 unique structures 2004:2023 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 8 1 theme heretofore-unavailable 1844:1865 arg1 level 1867:1871 a heretofore-unavailable level 1842:1871 a heretofore-unavailable level of analytical detail to glycan structure analysis 1842:1921 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 5 2 theme ion 1364:1366 arg1 spectrometer 1378:1389 a single ion trap mass spectrometer 1355:1389 a single ion trap mass spectrometer 1355:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 7 3 dep de 1682:1683 arg1 novo 1685:1688 novo 1685:1688 novo 1685:1688 The application of this approach exploits both library matching with standard materials and de novo approaches. 25048706 5 4 theme mass 1373:1376 arg1 spectrometer 1378:1389 a single ion trap mass spectrometer 1355:1389 a single ion trap mass spectrometer 1355:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 6 5 theme compositions 1495:1506 arg1 several 1509:1515 several 1509:1515 several 1509:1515 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 1 6 theme neonatal 239:246 arg1 N155 257:260 N155 257:260 N155 257:260 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 6 theme neonatal 239:246 arg1 strains 248:254 human neonatal strains 233:254 human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) 233:286 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 6 theme neonatal 239:246 arg1 RV3 276:278 RV3 276:278 RV3 276:278 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 3 7 theme ionization-mass 921:935 arg1 spectrometry 937:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 2 8 from library 567:573 arg1 identification 497:510 the identification 493:510 the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 493:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 8 9 theme detail 1887:1892 arg1 level 1867:1871 a heretofore-unavailable level 1842:1871 a heretofore-unavailable level of analytical detail to glycan structure analysis 1842:1921 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 1 10 theme milk 371:374 arg1 glycans 376:382 human milk glycans 365:382 human milk glycans 365:382 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 11 theme VP8 161:163 arg1 * 164:164 VP8* 161:164 VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 161:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 6 12 contain contained 1526:1534 arg2 mixtures 1545:1552 isomeric mixtures 1536:1552 isomeric mixtures including several novel structures 1536:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 6 12 contain contained 1526:1534 arg1 several 1509:1515 several 1509:1515 several 1509:1515 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 6 12 contain contained 1526:1534 arg2 structures 1578:1587 several novel structures 1564:1587 several novel structures 1564:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 10 13 theme glycans 2293:2299 arg1 glycans 2293:2299 glycans 2293:2299 glycans comprising the human milk metaglycome 2293:2337 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 10 13 theme glycans 2293:2299 arg1 repertoire 2279:2288 the repertoire 2275:2288 the repertoire of glycans comprising the human milk metaglycome 2275:2337 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 8 14 theme glycan 1897:1902 arg1 analysis 1914:1921 glycan structure analysis 1897:1921 glycan structure analysis 1897:1921 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 1 15 theme domains 166:172 arg1 forms 152:156 recombinant forms 140:156 recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 140:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 10 16 theme glycans 2233:2239 arg1 analysis 2210:2217 The functional glycomics analysis 2185:2217 The functional glycomics analysis of human milk glycans 2185:2239 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 10 17 theme milk 2322:2325 arg1 metaglycome 2327:2337 the human milk metaglycome 2312:2337 the human milk metaglycome 2312:2337 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 1 18 theme rotavirus 187:195 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 2 19 theme shotgun 455:461 arg1 microarray 470:479 a human milk glycan shotgun glycan microarray 435:479 a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 435:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 6 20 theme isomeric 1536:1543 arg1 structures 1578:1587 several novel structures 1564:1587 several novel structures 1564:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 6 20 theme isomeric 1536:1543 arg1 mixtures 1545:1552 isomeric mixtures 1536:1552 isomeric mixtures including several novel structures 1536:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 3 21 theme anti-glycan 739:749 arg1 antibody 751:758 anti-glycan antibody 739:758 anti-glycan antibody 739:758 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 2 22 theme milk 443:446 arg1 microarray 470:479 a human milk glycan shotgun glycan microarray 435:479 a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 435:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 4 23 theme glycan 1109:1114 arg1 library 1116:1122 the tagged glycan library 1098:1122 the tagged glycan library 1098:1122 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 3 24 theme metadata 677:684 arg1 metadata 677:684 metadata 677:684 metadata 677:684 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 3 24 theme metadata 677:684 arg1 variety 666:672 a variety 664:672 a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions 664:826 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 2 25 theme glycan 560:565 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 8 26 theme lectin 1761:1766 arg1 data 1789:1792 lectin and antibody-binding data 1761:1792 data 1789:1792 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 4 27 theme targets 1070:1076 arg1 targets 1070:1076 the glycan targets 1059:1076 the glycan targets 1059:1076 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 4 27 theme targets 1070:1076 arg1 28 1053:1054 28 1053:1054 28 1053:1054 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 8 28 theme antibody-binding 1772:1787 arg1 data 1789:1792 lectin and antibody-binding data 1761:1792 data 1789:1792 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 3 29 theme glycan 707:712 arg1 structures 714:723 the recognized glycan structures 692:723 the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions 692:826 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 0 30 theme human 69:73 arg1 glycans 80:86 human milk glycans 69:86 human milk glycans 69:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 2 31 theme glycans 527:533 arg1 identification 497:510 the identification 493:510 the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 493:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 4 32 theme glycans 1180:1186 arg1 disassembly 1165:1175 sequential disassembly 1154:1175 sequential disassembly of glycans by ion-trap mass spectrometry 1154:1216 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 0 33 theme glycans 80:86 arg1 spectrometry 47:58 multistage mass spectrometry 31:58 multistage mass spectrometry (MSn) of human milk glycans 31:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 0 33 theme glycans 80:86 arg1 MSn 61:63 MSn 61:63 MSn 61:63 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 3 34 theme specific 796:803 arg1 digestions 817:826 specific glycosidase digestions 796:826 specific glycosidase digestions 796:826 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 10 35 theme glycomics 2200:2208 arg1 analysis 2210:2217 The functional glycomics analysis 2185:2217 The functional glycomics analysis of human milk glycans 2185:2239 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 2 36 theme milk 548:551 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 0 37 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans 0:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 6 38 theme novel 1572:1576 arg1 structures 1578:1587 several novel structures 1564:1587 several novel structures 1564:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 6 39 from complexities 1445:1456 arg1 compositions 1495:1506 multiple compositions 1486:1506 multiple compositions 1486:1506 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 10 40 theme milk 2228:2231 arg1 glycans 2233:2239 human milk glycans 2222:2239 human milk glycans 2222:2239 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 9 41 theme glycan 1979:1984 arg1 targets 1986:1992 the 28 glycan targets 1972:1992 the 28 glycan targets analyzed 1972:2001 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 7 42 theme approach 1614:1621 arg1 application 1594:1604 The application 1590:1604 The application of this approach 1590:1621 The application of this approach exploits both library matching with standard materials and de novo approaches. 25048706 5 43 theme deeper 1242:1247 arg1 understanding 1260:1272 a deeper structural understanding 1240:1272 a deeper structural understanding of these key glycans 1240:1293 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 0 44 theme mass 42:45 arg1 spectrometry 47:58 multistage mass spectrometry 31:58 multistage mass spectrometry (MSn) of human milk glycans 31:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 0 44 theme mass 42:45 arg1 MSn 61:63 MSn 61:63 MSn 61:63 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 9 45 theme human 2056:2060 arg1 glycans 2067:2073 the human milk glycans 2052:2073 the human milk glycans recognized by human rotaviruses 2052:2105 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 3 46 theme compositional 840:852 arg1 information 854:864 compositional information 840:864 compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry 840:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 2 47 from identification 497:510 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 9 48 theme glycans 2067:2073 arg1 glycans 2067:2073 the human milk glycans 2052:2073 the human milk glycans recognized by human rotaviruses 2052:2105 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 9 48 theme glycans 2067:2073 arg1 23 2046:2047 23 2046:2047 23 2046:2047 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 5 49 theme structural 1328:1337 arg1 confirmation 1339:1350 structural confirmation 1328:1350 structural confirmation in a single ion trap mass spectrometer 1328:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 1 50 theme outer 197:201 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 3 51 theme matrix-assisted 888:902 arg1 spectrometry 937:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 4 52 theme metadata 1026:1033 arg1 analyses 1014:1021 analyses 1014:1021 analyses of metadata 1014:1033 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 4 53 theme mass 1200:1203 arg1 spectrometry 1205:1216 ion-trap mass spectrometry 1191:1216 ion-trap mass spectrometry 1191:1216 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 5 54 theme glycans 1287:1293 arg1 understanding 1260:1272 a deeper structural understanding 1240:1272 a deeper structural understanding of these key glycans 1240:1293 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 6 55 theme disassembly 1412:1422 arg1 strategy 1424:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 1 56 theme spike 210:214 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 6 57 theme sequential 1397:1406 arg1 strategy 1424:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 1 58 theme VP4 224:226 arg1 domains 166:172 VP8* domains 161:172 VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 161:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 58 theme VP4 224:226 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 8 59 theme structure 1904:1912 arg1 analysis 1914:1921 glycan structure analysis 1897:1921 glycan structure analysis 1897:1921 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 9 60 theme novel 2117:2121 arg1 structures 2123:2132 novel structures 2117:2132 novel structures not previously described as glycans in human milk 2117:2182 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 8 61 theme analytical 1876:1885 arg1 detail 1887:1892 analytical detail 1876:1892 analytical detail to glycan structure analysis 1876:1921 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 5 62 theme trap 1368:1371 arg1 spectrometer 1378:1389 a single ion trap mass spectrometer 1355:1389 a single ion trap mass spectrometer 1355:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 1 63 theme human 233:237 arg1 N155 257:260 N155 257:260 N155 257:260 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 63 theme human 233:237 arg1 strains 248:254 human neonatal strains 233:254 human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) 233:286 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 63 theme human 233:237 arg1 RV3 276:278 RV3 276:278 RV3 276:278 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 2 64 theme glycan 463:468 arg1 microarray 470:479 a human milk glycan shotgun glycan microarray 435:479 a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 435:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 3 65 theme desorption 910:919 arg1 spectrometry 937:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 1 66 theme human 365:369 arg1 glycans 376:382 human milk glycans 365:382 human milk glycans 365:382 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 6 67 theme multiple 1486:1493 arg1 compositions 1495:1506 multiple compositions 1486:1506 multiple compositions 1486:1506 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 4 68 theme glycan 1063:1068 arg1 targets 1070:1076 the glycan targets 1059:1076 the glycan targets 1059:1076 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 1 69 from strains 248:254 arg1 domains 166:172 VP8* domains 161:172 VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 161:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 69 from strains 248:254 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 69 from strains 248:254 arg1 forms 152:156 recombinant forms 140:156 recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 140:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 70 theme glycans 376:382 arg1 glycans 376:382 human milk glycans 365:382 human milk glycans 365:382 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 70 theme glycans 376:382 arg1 repertoire 351:360 the repertoire 347:360 the repertoire of human milk glycans 347:382 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 6 71 theme branching 1473:1481 arg1 complexities 1445:1456 the complexities 1441:1456 the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures 1441:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 1 72 theme * 164:164 arg1 domains 166:172 VP8* domains 161:172 VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 161:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 72 theme * 164:164 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 9 73 from glycans 2162:2168 arg1 milk 2179:2182 human milk 2173:2182 human milk 2173:2182 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 9 74 theme milk 2062:2065 arg1 glycans 2067:2073 the human milk glycans 2052:2073 the human milk glycans recognized by human rotaviruses 2052:2105 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 3 75 theme microarray 630:639 arg1 analyses 641:648 These microarray analyses 624:648 These microarray analyses 624:648 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 10 76 theme functional 2189:2198 arg1 analysis 2210:2217 The functional glycomics analysis 2185:2217 The functional glycomics analysis of human milk glycans 2185:2239 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 5 77 theme single 1357:1362 arg1 spectrometer 1378:1389 a single ion trap mass spectrometer 1355:1389 a single ion trap mass spectrometer 1355:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 6 78 theme linkage 1461:1467 arg1 complexities 1445:1456 the complexities 1441:1456 the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures 1441:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 9 79 theme studies 1945:1951 arg1 results 1928:1934 The results 1924:1934 The results of these studies 1924:1951 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 9 80 theme human 2173:2177 arg1 milk 2179:2182 human milk 2173:2182 human milk 2173:2182 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 1 81 theme human 181:185 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 10 82 theme human 2316:2320 arg1 metaglycome 2327:2337 the human milk metaglycome 2312:2337 the human milk metaglycome 2312:2337 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 4 83 theme tagged 1102:1107 arg1 library 1116:1122 the tagged glycan library 1098:1122 the tagged glycan library 1098:1122 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 2 84 theme glycan 448:453 arg1 microarray 470:479 a human milk glycan shotgun glycan microarray 435:479 a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 435:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 1 85 theme unique 325:330 arg1 glycans 332:338 unique glycans 325:338 unique glycans 325:338 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 4 86 theme sequential 1154:1163 arg1 disassembly 1165:1175 sequential disassembly 1154:1175 sequential disassembly of glycans by ion-trap mass spectrometry 1154:1216 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 2 87 theme human 437:441 arg1 microarray 470:479 a human milk glycan shotgun glycan microarray 435:479 a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 435:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 2 88 theme human 605:609 arg1 rotaviruses 611:621 these human rotaviruses 599:621 these human rotaviruses 599:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 2 89 from glycans 527:533 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 7 90 theme de 1682:1683 arg1 approaches 1690:1699 de novo approaches 1682:1699 de novo approaches 1682:1699 The application of this approach exploits both library matching with standard materials and de novo approaches. 25048706 0 91 theme milk 75:78 arg1 glycans 80:86 human milk glycans 69:86 human milk glycans 69:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 1 92 theme recombinant 140:150 arg1 forms 152:156 recombinant forms 140:156 recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 140:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 2 93 theme accompanying 389:400 arg1 study 402:406 The accompanying study 385:406 The accompanying study 385:406 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 1 94 dep strains 248:254 arg1 N155 257:260 N155 257:260 N155 257:260 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 94 dep strains 248:254 arg1 G3P[6 280:284 G3P[6 280:284 G3P[6 280:284 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 94 dep strains 248:254 arg1 RV3 276:278 RV3 276:278 RV3 276:278 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 94 dep strains 248:254 arg1 strains 248:254 human neonatal strains 233:254 human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) 233:286 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 94 dep strains 248:254 arg1 G10P[11 262:268 G10P[11 262:268 G10P[11 262:268 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 10 95 theme human 2222:2226 arg1 glycans 2233:2239 human milk glycans 2222:2239 human milk glycans 2222:2239 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 1 96 from forms 152:156 arg1 N155 257:260 N155 257:260 N155 257:260 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 96 from forms 152:156 arg1 RV3 276:278 RV3 276:278 RV3 276:278 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 96 from forms 152:156 arg1 strains 248:254 human neonatal strains 233:254 human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) 233:286 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 96 from forms 152:156 arg1 strain 301:306 a bovine strain 292:306 a bovine strain (B223) 292:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 96 from forms 152:156 arg1 B223 309:312 B223 309:312 B223 309:312 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 3 97 theme lectin 764:769 arg1 binding 771:777 lectin binding 764:777 lectin binding 764:777 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 2 98 theme specific 518:525 arg1 glycans 527:533 32 specific glycans 515:533 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses 515:621 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 5 99 theme structural 1249:1258 arg1 understanding 1260:1272 a deeper structural understanding 1240:1272 a deeper structural understanding of these key glycans 1240:1293 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 3 100 theme recognized 696:705 arg1 structures 714:723 the recognized glycan structures 692:723 the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions 692:826 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 0 101 theme human 102:106 arg1 rotaviruses 108:118 human rotaviruses 102:118 human rotaviruses 102:118 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 7 102 theme standard 1659:1666 arg1 materials 1668:1676 standard materials 1659:1676 standard materials 1659:1676 The application of this approach exploits both library matching with standard materials and de novo approaches. 25048706 3 103 theme glycosidase 805:815 arg1 digestions 817:826 specific glycosidase digestions 796:826 specific glycosidase digestions 796:826 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 3 104 from analysis 876:883 arg1 information 854:864 compositional information 840:864 compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry 840:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 4 105 theme detailed 1128:1135 arg1 sequencing 1137:1146 detailed sequencing 1128:1146 detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry 1128:1216 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 1 106 from strain 301:306 arg1 domains 166:172 VP8* domains 161:172 VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 161:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 106 from strain 301:306 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 106 from strain 301:306 arg1 forms 152:156 recombinant forms 140:156 recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) 140:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 107 theme bovine 294:299 arg1 B223 309:312 B223 309:312 B223 309:312 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 1 107 theme bovine 294:299 arg1 strain 301:306 a bovine strain 292:306 a bovine strain (B223) 292:313 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 8 108 theme glycosidase 1811:1821 arg1 digestions 1823:1832 glycosidase digestions 1811:1832 glycosidase digestions 1811:1832 This combination together with the metadata generated from lectin and antibody-binding data before and after glycosidase digestions provide a heretofore-unavailable level of analytical detail to glycan structure analysis. 25048706 2 109 theme human 542:546 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 0 110 theme multistage 31:40 arg1 spectrometry 47:58 multistage mass spectrometry 31:58 multistage mass spectrometry (MSn) of human milk glycans 31:86 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 0 110 theme multistage 31:40 arg1 MSn 61:63 MSn 61:63 MSn 61:63 Structural characterization by multistage mass spectrometry (MSn) of human milk glycans recognized by human rotaviruses. 25048706 6 111 theme several 1564:1570 arg1 structures 1578:1587 several novel structures 1564:1587 several novel structures 1564:1587 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 4 112 theme glycan 961:966 arg1 sequence 968:975 glycan sequence 961:975 glycan sequence 961:975 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 5 113 theme orthogonal 1304:1313 arg1 approach 1315:1322 an orthogonal approach 1301:1322 an orthogonal approach for structural confirmation in a single ion trap mass spectrometer 1301:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 10 114 theme significant 2250:2260 arg1 insight 2262:2268 significant insight 2250:2268 significant insight into the repertoire of glycans comprising the human milk metaglycome 2250:2337 The functional glycomics analysis of human milk glycans provides significant insight into the repertoire of glycans comprising the human milk metaglycome. 25048706 3 115 theme mass 871:874 arg1 analysis 876:883 mass analysis 871:883 mass analysis by matrix-assisted laser desorption ionization-mass spectrometry 871:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 4 116 theme ion-trap 1191:1198 arg1 spectrometry 1205:1216 ion-trap mass spectrometry 1191:1216 ion-trap mass spectrometry 1191:1216 To deduce glycan sequence and utilize information predicted by analyses of metadata from each glycan, 28 of the glycan targets were retrieved from the tagged glycan library for detailed sequencing using sequential disassembly of glycans by ion-trap mass spectrometry. 25048706 2 117 dep Yu 411:412 arg1 al. 417:419 Yu et al. 411:419 Yu et al.(2) 411:422 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 2 118 theme tagged 553:558 arg1 library 567:573 the human milk tagged glycan library 538:573 the human milk tagged glycan library 538:573 The accompanying study by Yu et al.(2), describes a human milk glycan shotgun glycan microarray that led to the identification of 32 specific glycans in the human milk tagged glycan library that were recognized by these human rotaviruses. 25048706 5 119 from confirmation 1339:1350 arg1 spectrometer 1378:1389 a single ion trap mass spectrometer 1355:1389 a single ion trap mass spectrometer 1355:1389 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 5 120 theme key 1283:1285 arg1 glycans 1287:1293 these key glycans 1277:1293 these key glycans 1277:1293 Our aim is to obtain a deeper structural understanding of these key glycans using an orthogonal approach for structural confirmation in a single ion trap mass spectrometer. 25048706 1 121 theme capsid 203:208 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25048706 3 122 theme laser 904:908 arg1 spectrometry 937:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 matrix-assisted laser desorption ionization-mass spectrometry 888:948 These microarray analyses also provided a variety of metadata about the recognized glycan structures compiled from anti-glycan antibody and lectin binding before and after specific glycosidase digestions, along with compositional information from mass analysis by matrix-assisted laser desorption ionization-mass spectrometry. 25048706 9 123 theme human 2089:2093 arg1 rotaviruses 2095:2105 human rotaviruses 2089:2105 human rotaviruses 2089:2105 The results of these studies showed that, among the 28 glycan targets analyzed, 27 unique structures were identified, and 23 of the human milk glycans recognized by human rotaviruses represent novel structures not previously described as glycans in human milk. 25048706 6 124 theme ion 1408:1410 arg1 strategy 1424:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy 1392:1431 This sequential ion disassembly strategy details the complexities of linkage and branching in multiple compositions, several of which contained isomeric mixtures including several novel structures. 25048706 1 125 theme protein 216:222 arg1 VP4 224:226 the human rotavirus outer capsid spike protein VP4 177:226 the human rotavirus outer capsid spike protein VP4 177:226 We have shown that recombinant forms of VP8* domains of the human rotavirus outer capsid spike protein VP4 from human neonatal strains (N155(G10P[11]) and RV3(G3P[6]) and a bovine strain (B223) recognize unique glycans within the repertoire of human milk glycans. 25658763 7 0 theme monocytes/macrophages 1605:1625 arg1 activities 1587:1596 inflammatory activities 1574:1596 inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression) 1574:1771 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 2 1 contain have 443:446 arg1 heterogeneousness 385:401 high-level heterogeneousness 374:401 high-level heterogeneousness of protein glycosylation of EMMPRIN 374:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 2 1 contain have 443:446 arg2 relevance 459:467 functional relevance 448:467 functional relevance to its biological effects 448:493 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 4 2 theme endothelial 1010:1020 arg1 cells 1022:1026 endothelial cells 1010:1026 endothelial cells 1010:1026 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 3 theme EMMPRIN 1291:1297 arg1 Glycosylation 1274:1286 Glycosylation 1274:1286 Glycosylation of EMMPRIN in monocytes/macrophages 1274:1322 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 8 4 theme biological 1875:1884 arg1 effects 1886:1892 the biological effects 1871:1892 the biological effects of EMMPRIN in monocytes/macrophages 1871:1928 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 7 5 dep activation 1634:1643 arg1 i.e. 1628:1631 i.e. 1628:1631 i.e. 1628:1631 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 6 theme inflammatory 1574:1585 arg1 activities 1587:1596 inflammatory activities 1574:1596 inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression) 1574:1771 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 3 7 theme extracellular 678:690 arg1 stimuli 692:698 different extracellular stimuli 668:698 different extracellular stimuli 668:698 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 4 8 theme cytokine 1046:1053 arg1 expression 1055:1064 cytokine expression 1046:1064 cytokine expression 1046:1064 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 9 theme less-glycosylated 1433:1449 arg1 form 1451:1454 the less-glycosylated form 1429:1454 the less-glycosylated form (LG) 1429:1459 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 6 9 theme less-glycosylated 1433:1449 arg1 LG 1457:1458 LG 1457:1458 LG 1457:1458 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 0 10 from effects 70:76 arg1 monocytes/macrophages 141:161 monocytes/macrophages 141:161 monocytes/macrophages 141:161 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 4 11 theme proteins 840:847 arg1 purification 794:805 the purification 790:805 the purification of highly- and less-glycosylated proteins respectively 790:860 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 12 gly Glycosylation 1274:1286 arg1 EMMPRIN 1291:1297 EMMPRIN 1291:1297 EMMPRIN in monocytes/macrophages 1291:1322 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 7 13 theme matrix 1733:1738 arg1 metalloproteinase 1740:1756 matrix metalloproteinase -9 1733:1759 matrix metalloproteinase -9 expression 1733:1770 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 6 14 theme complex-type 1404:1415 arg1 glycans 1417:1423 complex-type glycans 1404:1423 complex-type glycans 1404:1423 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 4 15 theme pathway 1109:1115 arg1 adhesion 998:1005 cell adhesion 993:1005 cell adhesion to endothelial cells 993:1026 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 15 theme pathway 1109:1115 arg1 migrations 1034:1043 cell migrations 1029:1043 cell migrations 1029:1043 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 15 theme pathway 1109:1115 arg1 expression 1055:1064 cytokine expression 1046:1064 cytokine expression 1046:1064 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 15 theme pathway 1109:1115 arg1 activation 1071:1080 activation 1071:1080 activation of inflammatory signalling pathway 1071:1115 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 8 16 dep CONCLUSIONS 1774:1784 arg1 represents 1821:1830 represents 1821:1830 represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages 1821:1928 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 7 17 dep HG-EMMPRIN 1490:1499 arg1 3 1478:1478 3 1478:1478 3 1478:1478 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 9 18 theme EMMPRIN 1948:1954 arg1 Glycosylation 1931:1943 Glycosylation 1931:1943 Glycosylation of EMMPRIN 1931:1954 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 9 18 theme EMMPRIN 1948:1954 arg1 target 1981:1986 a potential target 1969:1986 a potential target for regulating the inflammatory activities of monocytes/macrophages 1969:2054 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 0 19 theme extracellular 81:93 arg1 metalloproteinase 102:118 extracellular matrix metalloproteinase inducer (EMMPRIN) 81:136 extracellular matrix metalloproteinase inducer (EMMPRIN) 81:136 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 1 20 theme Extracellular 189:201 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 20 theme Extracellular 189:201 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 7 21 theme monocyte-endothelium 1683:1702 arg1 adhesion 1704:1711 enhanced monocyte-endothelium adhesion 1674:1711 enhanced monocyte-endothelium adhesion 1674:1711 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 4 22 theme forms 902:906 arg1 impacts 867:873 the impacts 863:873 the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN 863:949 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 0 23 theme metalloproteinase 102:118 arg1 effects 70:76 the pro-inflammatory effects 49:76 the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages 49:161 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 4 24 theme inflammatory 1085:1096 arg1 pathway 1109:1115 inflammatory signalling pathway 1085:1115 inflammatory signalling pathway 1085:1115 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 0 25 dep metalloproteinase 102:118 arg1 inducer 120:126 inducer 120:126 extracellular matrix metalloproteinase inducer (EMMPRIN) 81:136 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 0 25 dep metalloproteinase 102:118 arg1 EMMPRIN 129:135 EMMPRIN 129:135 EMMPRIN 129:135 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 0 26 theme Functional 0:9 arg1 relevance 11:19 Functional relevance 0:19 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages 0:161 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 4 27 from impacts 867:873 arg1 effects 932:938 the pro-inflammatory effects 911:938 the pro-inflammatory effects of EMMPRIN 911:949 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 2 28 theme inflammatory 510:521 arg1 activity 523:530 the inflammatory activity 506:530 the inflammatory activity of monocytes/macrophages 506:555 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 4 29 theme different 878:886 arg1 forms 902:906 different glycosylation forms 878:906 different glycosylation forms 878:906 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 9 30 theme inflammatory 2007:2018 arg1 activities 2020:2029 the inflammatory activities 2003:2029 the inflammatory activities of monocytes/macrophages 2003:2054 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 0 31 theme glycosylation 32:44 arg1 relevance 11:19 Functional relevance 0:19 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages 0:161 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 3 32 theme glycosylation 747:759 arg1 forms 761:765 different glycosylation forms 737:765 different glycosylation forms 737:765 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 4 33 theme various 968:974 arg1 adhesion 998:1005 cell adhesion 993:1005 cell adhesion to endothelial cells 993:1026 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 33 theme various 968:974 arg1 aspects 976:982 various aspects 968:982 various aspects 968:982 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 33 theme various 968:974 arg1 migrations 1034:1043 cell migrations 1029:1043 cell migrations 1029:1043 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 33 theme various 968:974 arg1 expression 1055:1064 cytokine expression 1046:1064 cytokine expression 1046:1064 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 33 theme various 968:974 arg1 activation 1071:1080 activation 1071:1080 activation of inflammatory signalling pathway 1071:1115 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 2 34 theme glycosylation 414:426 arg1 heterogeneousness 385:401 high-level heterogeneousness 374:401 high-level heterogeneousness of protein glycosylation of EMMPRIN 374:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 3 35 theme glycosylation 570:582 arg1 patterns 584:591 The glycosylation patterns 566:591 The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli 566:698 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 8 36 theme Post-transcriptional 1786:1805 arg1 glycosylation 1807:1819 Post-transcriptional glycosylation 1786:1819 Post-transcriptional glycosylation 1786:1819 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 1 37 theme important 252:260 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 37 theme important 252:260 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 4 38 theme EMMPRIN 943:949 arg1 effects 932:938 the pro-inflammatory effects 911:938 the pro-inflammatory effects of EMMPRIN 911:949 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 2 39 theme EMMPRIN 431:437 arg1 glycosylation 414:426 protein glycosylation 406:426 protein glycosylation of EMMPRIN 406:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 8 40 theme important 1835:1843 arg1 mechanism 1845:1853 an important mechanism 1832:1853 an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages 1832:1928 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 4 41 theme cell 993:996 arg1 adhesion 998:1005 cell adhesion 993:1005 cell adhesion to endothelial cells 993:1026 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 42 theme HG 1385:1386 arg1 form 1388:1391 the HG form 1381:1391 the HG form containing complex-type glycans 1381:1423 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 1 43 theme cellular 303:310 arg1 functions 312:320 the cellular functions 299:320 the cellular functions of monocytes/macrophages 299:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 3 44 theme different 668:676 arg1 stimuli 692:698 different extracellular stimuli 668:698 different extracellular stimuli 668:698 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 7 45 theme pro-inflammatory 1534:1549 arg1 effects 1551:1557 pro-inflammatory effects 1534:1557 pro-inflammatory effects 1534:1557 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 3 46 theme THP-1 640:644 arg1 monocytes/macrophages 617:637 monocytes/macrophages 617:637 monocytes/macrophages (THP-1 cells) 617:651 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 3 46 theme THP-1 640:644 arg1 cells 646:650 THP-1 cells 640:650 THP-1 cells 640:650 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 1 47 theme matrix 203:208 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 47 theme matrix 203:208 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 6 48 dep form 1388:1391 arg1 type 1472:1475 the simple type 1461:1475 the simple type 1461:1475 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 5 49 dep form 1167:1170 arg1 1 1126:1126 1 1126:1126 1 1126:1126 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 2 50 theme high-level 374:383 arg1 heterogeneousness 385:401 high-level heterogeneousness 374:401 high-level heterogeneousness of protein glycosylation of EMMPRIN 374:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 3 51 dep observed 705:712 arg1 METHODS 558:564 METHODS 558:564 METHODS 558:564 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 4 52 gly less-glycosylated 822:838 arg1 proteins 840:847 highly- and less-glycosylated proteins 810:847 highly- and less-glycosylated proteins respectively 810:860 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 8 53 theme EMMPRIN 1897:1903 arg1 effects 1886:1892 the biological effects 1871:1892 the biological effects of EMMPRIN in monocytes/macrophages 1871:1928 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 2 54 theme protein 406:412 arg1 glycosylation 414:426 protein glycosylation 406:426 protein glycosylation of EMMPRIN 406:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 6 55 gly less-glycosylated 1433:1449 arg1 form 1451:1454 the less-glycosylated form 1429:1454 the less-glycosylated form (LG) 1429:1459 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 6 55 gly less-glycosylated 1433:1449 arg1 LG 1457:1458 LG 1457:1458 LG 1457:1458 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 7 56 theme metalloproteinase 1740:1756 arg1 expression 1761:1770 matrix metalloproteinase -9 expression 1733:1770 matrix metalloproteinase -9 expression 1733:1770 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 57 theme Only 1481:1484 arg1 HG-EMMPRIN 1490:1499 the HG-EMMPRIN 1486:1499 3) Only the HG-EMMPRIN but not the LG-EMMPRIN 1478:1522 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 9 58 gly Glycosylation 1931:1943 arg1 EMMPRIN 1948:1954 EMMPRIN 1948:1954 EMMPRIN 1948:1954 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 4 59 theme less-glycosylated 822:838 arg1 proteins 840:847 highly- and less-glycosylated proteins 810:847 highly- and less-glycosylated proteins respectively 810:860 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 5 60 theme EMMPRIN 1175:1181 arg1 It 1129:1130 It 1129:1130 It 1129:1130 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 60 theme EMMPRIN 1175:1181 arg1 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 60 theme EMMPRIN 1175:1181 arg1 form 1167:1170 the highly-glycosylated form 1143:1170 the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2) 1143:1268 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 7 61 theme cell 1714:1717 arg1 migration 1719:1727 cell migration 1714:1727 cell migration 1714:1727 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 4 62 theme cell 1029:1032 arg1 migrations 1034:1043 cell migrations 1029:1043 cell migrations 1029:1043 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 63 dep led 1324:1326 arg1 2 1271:1271 2 1271:1271 2 1271:1271 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 1 64 theme BACKGROUND 164:173 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 64 theme BACKGROUND 164:173 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 4 65 theme signalling 1098:1107 arg1 pathway 1109:1115 inflammatory signalling pathway 1085:1115 inflammatory signalling pathway 1085:1115 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 66 from monocytes/macrophages 1302:1322 arg1 Glycosylation 1274:1286 Glycosylation 1274:1286 Glycosylation of EMMPRIN in monocytes/macrophages 1274:1322 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 6 67 theme simple 1465:1470 arg1 type 1472:1475 the simple type 1461:1475 the simple type 1461:1475 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 0 68 theme matrix 95:100 arg1 metalloproteinase 102:118 extracellular matrix metalloproteinase inducer (EMMPRIN) 81:136 extracellular matrix metalloproteinase inducer (EMMPRIN) 81:136 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 1 69 theme OBJECTIVE 179:187 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 69 theme OBJECTIVE 179:187 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 7 70 theme enhanced 1674:1681 arg1 adhesion 1704:1711 enhanced monocyte-endothelium adhesion 1674:1711 enhanced monocyte-endothelium adhesion 1674:1711 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 4 71 theme highly- 810:816 arg1 proteins 840:847 highly- and less-glycosylated proteins 810:847 highly- and less-glycosylated proteins respectively 810:860 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 4 72 theme glycosylation 888:900 arg1 forms 902:906 different glycosylation forms 878:906 different glycosylation forms 878:906 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 6 73 from Glycosylation 1274:1286 arg1 monocytes/macrophages 1302:1322 monocytes/macrophages 1302:1322 monocytes/macrophages 1302:1322 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 9 74 theme potential 1971:1979 arg1 target 1981:1986 a potential target 1969:1986 a potential target for regulating the inflammatory activities of monocytes/macrophages 1969:2054 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 9 74 theme potential 1971:1979 arg1 Glycosylation 1931:1943 Glycosylation 1931:1943 Glycosylation of EMMPRIN 1931:1954 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 7 75 dep ERK1/2 1648:1653 arg1 pathway 1665:1671 pathway 1665:1671 pathway 1665:1671 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 6 76 from EMMPRIN 1291:1297 arg1 monocytes/macrophages 1302:1322 monocytes/macrophages 1302:1322 monocytes/macrophages 1302:1322 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 5 77 theme inflammatory 1234:1245 arg1 H2O2 1264:1267 H2O2 1264:1267 H2O2 1264:1267 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 77 theme inflammatory 1234:1245 arg1 PMA 1256:1258 PMA 1256:1258 PMA 1256:1258 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 77 theme inflammatory 1234:1245 arg1 signals 1247:1253 inflammatory signals 1234:1253 inflammatory signals (PMA and H2O2) 1234:1268 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 6 78 contain containing 1393:1402 arg1 form 1388:1391 the HG form 1381:1391 the HG form containing complex-type glycans 1381:1423 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 6 78 contain containing 1393:1402 arg2 glycans 1417:1423 complex-type glycans 1404:1423 complex-type glycans 1404:1423 2) Glycosylation of EMMPRIN in monocytes/macrophages led to N-linked-glycans being added to the protein, with the HG form containing complex-type glycans and the less-glycosylated form (LG) the simple type. 25658763 2 79 theme functional 448:457 arg1 relevance 459:467 functional relevance 448:467 functional relevance to its biological effects 448:493 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 0 80 theme protein 24:30 arg1 glycosylation 32:44 protein glycosylation 24:44 protein glycosylation 24:44 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 1 81 theme monocytes/macrophages 325:345 arg1 functions 312:320 the cellular functions 299:320 the cellular functions of monocytes/macrophages 299:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 9 82 theme monocytes/macrophages 2034:2054 arg1 activities 2020:2029 the inflammatory activities 2003:2029 the inflammatory activities of monocytes/macrophages 2003:2054 Glycosylation of EMMPRIN may serve as a potential target for regulating the inflammatory activities of monocytes/macrophages. 25658763 5 83 gly highly-glycosylated 1147:1165 arg1 It 1129:1130 It 1129:1130 It 1129:1130 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 83 gly highly-glycosylated 1147:1165 arg1 HG-EMMPRIN 1184:1193 HG-EMMPRIN 1184:1193 HG-EMMPRIN 1184:1193 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 83 gly highly-glycosylated 1147:1165 arg1 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 83 gly highly-glycosylated 1147:1165 arg1 EMMPRIN 1175:1181 EMMPRIN 1175:1181 EMMPRIN (HG-EMMPRIN) 1175:1194 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 83 gly highly-glycosylated 1147:1165 arg1 form 1167:1170 the highly-glycosylated form 1143:1170 the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2) 1143:1268 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 84 dep signals 1247:1253 arg1 H2O2 1264:1267 H2O2 1264:1267 H2O2 1264:1267 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 84 dep signals 1247:1253 arg1 PMA 1256:1258 PMA 1256:1258 PMA 1256:1258 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 84 dep signals 1247:1253 arg1 signals 1247:1253 inflammatory signals 1234:1253 inflammatory signals (PMA and H2O2) 1234:1268 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 3 85 theme forms 761:765 arg1 structures 723:732 the structures 719:732 the structures of different glycosylation forms 719:765 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 3 86 gly glycosylation 570:582 arg1 EMMPRIN 596:602 EMMPRIN 596:602 EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli 596:698 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 7 87 theme NF-κB 1659:1663 arg1 activation 1634:1643 activation 1634:1643 activation of ERK1/2 and NF-κB pathway 1634:1671 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 87 theme NF-κB 1659:1663 arg1 adhesion 1704:1711 enhanced monocyte-endothelium adhesion 1674:1711 enhanced monocyte-endothelium adhesion 1674:1711 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 87 theme NF-κB 1659:1663 arg1 expression 1761:1770 matrix metalloproteinase -9 expression 1733:1770 matrix metalloproteinase -9 expression 1733:1770 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 87 theme NF-κB 1659:1663 arg1 migration 1719:1727 cell migration 1714:1727 cell migration 1714:1727 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 3 88 theme different 737:745 arg1 forms 761:765 different glycosylation forms 737:765 different glycosylation forms 737:765 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 8 89 from effects 1886:1892 arg1 monocytes/macrophages 1908:1928 monocytes/macrophages 1908:1928 monocytes/macrophages 1908:1928 CONCLUSIONS Post-transcriptional glycosylation represents an important mechanism that determines the biological effects of EMMPRIN in monocytes/macrophages. 25658763 2 90 theme biological 476:485 arg1 effects 487:493 its biological effects 472:493 its biological effects 472:493 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 0 91 theme pro-inflammatory 53:68 arg1 effects 70:76 the pro-inflammatory effects 49:76 the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages 49:161 Functional relevance of protein glycosylation to the pro-inflammatory effects of extracellular matrix metalloproteinase inducer (EMMPRIN) on monocytes/macrophages. 25658763 7 92 theme ERK1/2 1648:1653 arg1 activation 1634:1643 activation 1634:1643 activation of ERK1/2 and NF-κB pathway 1634:1671 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 92 theme ERK1/2 1648:1653 arg1 adhesion 1704:1711 enhanced monocyte-endothelium adhesion 1674:1711 enhanced monocyte-endothelium adhesion 1674:1711 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 92 theme ERK1/2 1648:1653 arg1 expression 1761:1770 matrix metalloproteinase -9 expression 1733:1770 matrix metalloproteinase -9 expression 1733:1770 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 7 92 theme ERK1/2 1648:1653 arg1 migration 1719:1727 cell migration 1714:1727 cell migration 1714:1727 3) Only the HG-EMMPRIN but not the LG-EMMPRIN exhibited pro-inflammatory effects and stimulated inflammatory activities of the monocytes/macrophages (i.e., activation of ERK1/2 and NF-κB pathway, enhanced monocyte-endothelium adhesion, cell migration and matrix metalloproteinase -9 expression). 25658763 5 93 theme highly-glycosylated 1147:1165 arg1 It 1129:1130 It 1129:1130 It 1129:1130 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 93 theme highly-glycosylated 1147:1165 arg1 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1118:1124 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 5 93 theme highly-glycosylated 1147:1165 arg1 form 1167:1170 the highly-glycosylated form 1143:1170 the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2) 1143:1268 RESULTS 1) It was mainly the highly-glycosylated form of EMMPRIN (HG-EMMPRIN) that increased after being exposed to inflammatory signals (PMA and H2O2). 25658763 2 94 theme monocytes/macrophages 535:555 arg1 activity 523:530 the inflammatory activity 506:530 the inflammatory activity of monocytes/macrophages 506:555 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 2 95 gly glycosylation 414:426 arg1 EMMPRIN 431:437 EMMPRIN 431:437 EMMPRIN 431:437 We have hypothesized that high-level heterogeneousness of protein glycosylation of EMMPRIN may have functional relevance to its biological effects and affect the inflammatory activity of monocytes/macrophages. 25658763 1 96 dep metalloproteinase 210:226 arg1 inducer 228:234 inducer 228:234 inducer (EMMPRIN) 228:244 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 96 dep metalloproteinase 210:226 arg1 EMMPRIN 237:243 EMMPRIN 237:243 EMMPRIN 237:243 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 3 97 theme EMMPRIN 596:602 arg1 patterns 584:591 The glycosylation patterns 566:591 The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli 566:698 METHODS The glycosylation patterns of EMMPRIN expressed by monocytes/macrophages (THP-1 cells) in response to different extracellular stimuli were observed, and the structures of different glycosylation forms were identified. 25658763 4 98 theme pro-inflammatory 915:930 arg1 effects 932:938 the pro-inflammatory effects 911:938 the pro-inflammatory effects of EMMPRIN 911:949 After the purification of highly- and less-glycosylated proteins respectively, the impacts of different glycosylation forms on the pro-inflammatory effects of EMMPRIN were examined in various aspects, such as cell adhesion to endothelial cells, cell migrations, cytokine expression, and activation of inflammatory signalling pathway. 25658763 1 99 theme pro-inflammatory 262:277 arg1 protein 279:285 an important pro-inflammatory protein 249:285 an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages 249:345 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 25658763 1 99 theme pro-inflammatory 262:277 arg1 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) 164:244 metalloproteinase 210:226 BACKGROUND AND OBJECTIVE Extracellular matrix metalloproteinase inducer (EMMPRIN) is an important pro-inflammatory protein involved in the cellular functions of monocytes/macrophages. 24965454 7 0 theme induced 1447:1453 arg1 hindrance 1462:1470 induced steric hindrance 1447:1470 induced steric hindrance upon primary receptor binding 1447:1500 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 3 1 theme vulnerability 671:683 arg1 vulnerability 671:683 vulnerability 671:683 vulnerability 671:683 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 3 1 theme vulnerability 671:683 arg1 supersite 658:666 a supersite 656:666 a supersite of vulnerability on gp120 656:692 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 6 2 theme glycan 1276:1281 arg1 knockouts 1283:1291 glycan knockouts 1276:1291 glycan knockouts 1276:1291 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 1 3 theme viral 265:269 arg1 challenge 271:279 viral challenge 265:279 viral challenge in animal models 265:296 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 6 4 theme high-resolution 1198:1212 arg1 structures 1222:1231 high-resolution crystal structures 1198:1231 high-resolution crystal structures 1198:1231 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 11 5 theme structural 2074:2083 arg1 data 2085:2088 structural data 2074:2088 structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness 2074:2235 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 5 6 dep soluble 1009:1015 arg1 trimeric 1018:1025 trimeric 1018:1025 trimeric 1018:1025 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 2 7 link N-linked 379:386 arg1 glycans 388:394 conserved N-linked glycans 369:394 conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332 369:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 3 8 theme 2G12 528:531 arg1 epitope 533:539 the 2G12 epitope 524:539 the 2G12 epitope 524:539 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 8 9 theme glycan 1677:1682 arg1 assignments 1684:1694 our low-resolution model and glycan assignments 1648:1694 assignments 1684:1694 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 4 10 theme 2G12 845:848 arg1 interaction 830:840 the interaction 826:840 the interaction of 2G12 with gp120 or the Env trimer 826:877 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 7 11 theme soluble 1377:1383 arg1 CD4 1385:1387 soluble CD4 1377:1387 soluble CD4 1377:1387 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 9 12 theme people 1873:1878 arg1 millions 1861:1868 millions 1861:1868 millions of people around the world 1861:1895 IMPORTANCE HIV-1 is a human virus that results in the deaths of millions of people around the world each year. 24965454 9 13 dep IMPORTANCE 1797:1806 arg1 HIV-1 1808:1812 HIV-1 1808:1812 IMPORTANCE HIV-1 1797:1812 IMPORTANCE HIV-1 is a human virus that results in the deaths of millions of people around the world each year. 24965454 2 14 theme unique 309:314 arg1 2G12 299:302 2G12 299:302 2G12 299:302 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 2 14 theme unique 309:314 arg1 antibody 334:341 a unique, domain-exchanged antibody 307:341 a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332 307:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 1 15 theme particular 194:203 arg1 interest 205:212 particular interest 194:212 particular interest 194:212 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 6 16 dep mapped 1147:1152 arg1 performing 1240:1249 performing 1240:1249 performing neutralization assays of glycan knockouts 1240:1291 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 6 16 dep mapped 1147:1152 arg1 fitting 1190:1196 fitting 1190:1196 fitting high-resolution crystal structures 1190:1231 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 0 17 theme electron 114:121 arg1 microscopy 123:132 negative-stain single-particle electron microscopy 83:132 negative-stain single-particle electron microscopy 83:132 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 2 18 from position 479:486 arg1 glycan 469:474 a glycan 467:474 a glycan at position N332 467:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 12 19 theme antibody-based 2393:2406 arg1 therapies 2408:2416 antibody-based therapies 2393:2416 antibody-based therapies 2393:2416 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 8 20 theme higher-resolution 1716:1732 arg1 studies 1734:1740 higher-resolution studies 1716:1740 higher-resolution studies 1716:1740 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 1 21 dep protection 237:246 arg1 it 248:249 it 248:249 it 248:249 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 1 21 dep protection 237:246 arg1 provides 251:258 provides 251:258 provides from viral challenge in animal models 251:296 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 7 22 theme complex 1348:1354 arg1 Å 1331:1331 ∼26 Å 1327:1331 ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env 1327:1396 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 4 23 theme crystal 701:707 arg1 structures 709:718 crystal structures 701:718 crystal structures of 2G12 and 2G12 bound to high-mannose glycans 701:765 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 10 24 theme several 1924:1930 arg1 therapeutics 1942:1953 several effective therapeutics 1924:1953 several effective therapeutics available to prolong life 1924:1979 While there are several effective therapeutics available to prolong life, a vaccine is the best long-term solution for curbing this global epidemic. 24965454 3 25 theme other 574:578 arg1 antibodies 609:618 other potent, broadly neutralizing antibodies 574:618 other potent, broadly neutralizing antibodies 574:618 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 1 26 theme UNLABELLED 135:144 arg1 antibody 174:181 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 11 27 theme first 2139:2143 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 5 28 gly glycosylated 1117:1128 arg1 epitope 1130:1136 its native and fully glycosylated epitope 1096:1136 its native and fully glycosylated epitope 1096:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 3 29 dep shown 551:555 arg1 constitutes 644:654 constitutes 644:654 constitutes a supersite of vulnerability on gp120 644:692 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 7 30 theme coreceptor 1545:1554 arg1 s 1556:1556 coreceptor(s) 1545:1557 coreceptor(s) 1545:1557 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 0 31 theme glycosylated 57:68 arg1 Env 76:78 soluble and fully glycosylated HIV-1 Env 39:78 soluble and fully glycosylated HIV-1 Env 39:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 4 32 with interaction 830:840 arg1 gp120 855:859 gp120 855:859 gp120 855:859 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 4 32 with interaction 830:840 arg1 trimer 872:877 the Env trimer 864:877 the Env trimer 864:877 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 0 33 from Structure 0:8 arg1 complex 26:32 complex 26:32 complex with soluble and fully glycosylated HIV-1 Env 26:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 4 34 theme high-mannose 746:757 arg1 glycans 759:765 high-mannose glycans 746:765 high-mannose glycans 746:765 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 8 35 theme epitope 1788:1794 arg1 nature 1769:1774 the molecular nature 1755:1774 the molecular nature of the 2G12 epitope 1755:1794 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 5 36 theme microscopy 952:961 arg1 reconstruction 963:976 a negative-stain single-particle electron microscopy reconstruction 910:976 a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 910:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 7 37 from Å 1331:1331 arg1 reconstruction 1309:1322 a reconstruction 1307:1322 a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env 1307:1396 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 11 38 theme antibodies 2164:2173 arg1 2G12 2185:2188 2G12 2185:2188 2G12 2185:2188 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 38 theme antibodies 2164:2173 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 38 theme antibodies 2164:2173 arg1 one 2128:2130 one 2128:2130 one 2128:2130 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 0 39 theme negative-stain 83:96 arg1 microscopy 123:132 negative-stain single-particle electron microscopy 83:132 negative-stain single-particle electron microscopy 83:132 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 1 40 theme animal 284:289 arg1 models 291:296 animal models 284:296 animal models 284:296 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 2 41 theme outer 442:446 arg1 domain 448:453 the gp120 outer domain 432:453 the gp120 outer domain centered on a glycan at position N332 432:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 5 42 theme single-particle 927:941 arg1 microscopy 952:961 a negative-stain single-particle electron microscopy 910:961 a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 910:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 5 43 theme glycosylated 1117:1128 arg1 epitope 1130:1136 its native and fully glycosylated epitope 1096:1136 its native and fully glycosylated epitope 1096:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 7 44 theme receptor 1485:1492 arg1 binding 1494:1500 primary receptor binding 1477:1500 primary receptor binding 1477:1500 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 11 45 theme binding 2112:2118 arg1 site 2120:2123 the viral binding site 2102:2123 the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12 2102:2188 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 45 theme binding 2112:2118 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 45 theme binding 2112:2118 arg1 one 2128:2130 one 2128:2130 one 2128:2130 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 12 46 theme 2G12 2342:2345 arg1 epitope 2347:2353 the 2G12 epitope 2338:2353 the 2G12 epitope 2338:2353 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 1 47 theme sterilizing 225:235 arg1 protection 237:246 the sterilizing protection it provides from viral challenge in animal models 221:296 the sterilizing protection it provides from viral challenge in animal models 221:296 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 7 48 theme steric 1455:1460 arg1 hindrance 1462:1470 induced steric hindrance 1447:1470 induced steric hindrance upon primary receptor binding 1447:1500 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 5 49 theme Fab2 986:989 arg1 reconstruction 963:976 a negative-stain single-particle electron microscopy reconstruction 910:976 a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 910:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 2 50 from patch 423:427 arg1 domain 448:453 the gp120 outer domain 432:453 the gp120 outer domain centered on a glycan at position N332 432:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 1 51 theme neutralizing 150:161 arg1 antibody 174:181 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 9 52 dep year 1902:1905 arg1 each 1897:1900 each 1897:1900 each 1897:1900 IMPORTANCE HIV-1 is a human virus that results in the deaths of millions of people around the world each year. 24965454 2 53 theme N-linked 379:386 arg1 glycans 388:394 conserved N-linked glycans 369:394 conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332 369:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 3 54 from supersite 658:666 arg1 gp120 688:692 gp120 688:692 gp120 688:692 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 6 55 theme knockouts 1283:1291 arg1 assays 1266:1271 neutralization assays 1251:1271 neutralization assays of glycan knockouts 1251:1291 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 7 56 theme membrane 1428:1435 arg1 fusion 1437:1442 membrane fusion 1428:1442 membrane fusion 1428:1442 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 6 57 theme relevant 1154:1161 arg1 glycans 1163:1169 relevant glycans 1154:1169 relevant glycans 1154:1169 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 6 58 theme neutralization 1251:1264 arg1 assays 1266:1271 neutralization assays 1251:1271 neutralization assays of glycan knockouts 1251:1291 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 8 59 theme low-resolution 1652:1665 arg1 model 1667:1671 our low-resolution model and glycan assignments 1648:1694 model 1667:1671 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 3 60 dep potent 580:585 arg1 neutralizing 596:607 neutralizing 596:607 neutralizing 596:607 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 1 61 from challenge 271:279 arg1 models 291:296 animal models 284:296 animal models 284:296 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 3 62 theme potent 580:585 arg1 antibodies 609:618 other potent, broadly neutralizing antibodies 574:618 other potent, broadly neutralizing antibodies 574:618 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 10 63 dep is 1992:1993 arg1 solution 2014:2021 the best long-term solution 1995:2021 the best long-term solution for curbing this global epidemic 1995:2054 While there are several effective therapeutics available to prolong life, a vaccine is the best long-term solution for curbing this global epidemic. 24965454 7 64 theme 2G12 1366:1369 arg1 Fab2 1371:1374 2G12 Fab2 1366:1374 2G12 Fab2 1366:1374 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 10 65 theme effective 1932:1940 arg1 therapeutics 1942:1953 several effective therapeutics 1924:1953 several effective therapeutics available to prolong life 1924:1979 While there are several effective therapeutics available to prolong life, a vaccine is the best long-term solution for curbing this global epidemic. 24965454 4 66 theme Env 868:870 arg1 trimer 872:877 the Env trimer 864:877 the Env trimer 864:877 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 5 67 with interaction 1079:1089 arg1 epitope 1130:1136 its native and fully glycosylated epitope 1096:1136 its native and fully glycosylated epitope 1096:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 7 68 with interaction 1528:1538 arg1 s 1556:1556 coreceptor(s) 1545:1557 coreceptor(s) 1545:1557 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 7 69 theme ternary 1340:1346 arg1 complex 1348:1354 the ternary complex 1336:1354 the ternary complex formed by 2G12 Fab2, soluble CD4, and Env 1336:1396 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 2 70 dep unique 309:314 arg1 domain-exchanged 317:332 domain-exchanged 317:332 domain-exchanged 317:332 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 5 71 theme ∼17-Å 1034:1038 arg1 resolution 1040:1049 ∼17-Å resolution 1034:1049 ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 1034:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 10 72 theme long-term 2004:2012 arg1 solution 2014:2021 the best long-term solution 1995:2021 the best long-term solution for curbing this global epidemic 1995:2054 While there are several effective therapeutics available to prolong life, a vaccine is the best long-term solution for curbing this global epidemic. 24965454 11 73 theme HIV-1-neutralizing 2145:2162 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 0 74 from Fab2 18:21 arg1 complex 26:32 complex 26:32 complex with soluble and fully glycosylated HIV-1 Env 26:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 10 75 theme available 1955:1963 arg1 therapeutics 1942:1953 several effective therapeutics 1924:1953 several effective therapeutics available to prolong life 1924:1979 While there are several effective therapeutics available to prolong life, a vaccine is the best long-term solution for curbing this global epidemic. 24965454 0 76 theme HIV-1 70:74 arg1 Env 76:78 soluble and fully glycosylated HIV-1 Env 39:78 soluble and fully glycosylated HIV-1 Env 39:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 5 77 theme native 1100:1105 arg1 epitope 1130:1136 its native and fully glycosylated epitope 1096:1136 its native and fully glycosylated epitope 1096:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 8 78 theme molecular 1759:1767 arg1 nature 1769:1774 the molecular nature 1755:1774 the molecular nature of the 2G12 epitope 1755:1794 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 1 79 theme anti-HIV-1 163:172 arg1 antibody 174:181 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 135:186 UNLABELLED The neutralizing anti-HIV-1 antibody 2G12 is of particular interest due to the sterilizing protection it provides from viral challenge in animal models. 24965454 5 80 theme electron 943:950 arg1 microscopy 952:961 a negative-stain single-particle electron microscopy 910:961 a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 910:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 0 81 theme single-particle 98:112 arg1 microscopy 123:132 negative-stain single-particle electron microscopy 83:132 negative-stain single-particle electron microscopy 83:132 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 8 82 theme 2G12 1783:1786 arg1 epitope 1788:1794 the 2G12 epitope 1779:1794 the 2G12 epitope 1779:1794 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 0 83 gly glycosylated 57:68 arg1 Env 76:78 soluble and fully glycosylated HIV-1 Env 39:78 soluble and fully glycosylated HIV-1 Env 39:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 0 84 from complex 26:32 arg1 Structure 0:8 Structure 0:8 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 0:133 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 12 85 theme molecular 2318:2326 arg1 nature 2328:2333 the molecular nature 2314:2333 the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies 2314:2416 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 5 86 from resolution 1040:1049 arg1 Env 1027:1029 a soluble, trimeric Env 1007:1029 a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 1007:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 11 87 theme viral 2106:2110 arg1 site 2120:2123 the viral binding site 2102:2123 the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12 2102:2188 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 87 theme viral 2106:2110 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 87 theme viral 2106:2110 arg1 one 2128:2130 one 2128:2130 one 2128:2130 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 4 88 theme 2G12 723:726 arg1 structures 709:718 crystal structures 701:718 crystal structures of 2G12 and 2G12 bound to high-mannose glycans 701:765 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 0 89 theme Fab2 18:21 arg1 Structure 0:8 Structure 0:8 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 0:133 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 5 90 theme negative-stain 912:925 arg1 microscopy 952:961 a negative-stain single-particle electron microscopy 910:961 a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 910:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 2 91 theme gp120 436:440 arg1 domain 448:453 the gp120 outer domain 432:453 the gp120 outer domain centered on a glycan at position N332 432:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 12 92 theme vaccine 2374:2380 arg1 design 2382:2387 vaccine design 2374:2387 vaccine design 2374:2387 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 4 93 theme 2G12 732:735 arg1 structures 709:718 crystal structures 701:718 crystal structures of 2G12 and 2G12 bound to high-mannose glycans 701:765 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 11 94 theme one 2128:2130 arg1 site 2120:2123 the viral binding site 2102:2123 the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12 2102:2188 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 94 theme one 2128:2130 arg1 antibodies 2164:2173 the first HIV-1-neutralizing antibodies 2135:2173 the first HIV-1-neutralizing antibodies isolated 2135:2182 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 11 94 theme one 2128:2130 arg1 one 2128:2130 one 2128:2130 one 2128:2130 Here, we present structural data that reveal the viral binding site of one of the first HIV-1-neutralizing antibodies isolated, 2G12, and provide a rationale for its effectiveness. 24965454 12 95 theme epitope 2347:2353 arg1 nature 2328:2333 the molecular nature 2314:2333 the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies 2314:2416 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 4 96 theme structural 788:797 arg1 information 799:809 no structural information 785:809 no structural information that describes the interaction of 2G12 with gp120 or the Env trimer 785:877 While crystal structures of 2G12 and 2G12 bound to high-mannose glycans have been solved, no structural information that describes the interaction of 2G12 with gp120 or the Env trimer is available. 24965454 2 97 theme high-mannose 410:421 arg1 patch 423:427 the high-mannose patch 406:427 the high-mannose patch on the gp120 outer domain centered on a glycan at position N332 406:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 0 98 theme soluble 39:45 arg1 Env 76:78 soluble and fully glycosylated HIV-1 Env 39:78 soluble and fully glycosylated HIV-1 Env 39:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 5 99 theme soluble 1009:1015 arg1 Env 1027:1029 a soluble, trimeric Env 1007:1029 a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 1007:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 12 100 theme higher-resolution 2275:2291 arg1 studies 2293:2299 higher-resolution studies 2275:2299 higher-resolution studies 2275:2299 These structures provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope, which will aid in vaccine design and antibody-based therapies. 24965454 3 101 theme Several 494:500 arg1 glycans 502:508 Several glycans 494:508 Several glycans in and around the 2G12 epitope 494:539 Several glycans in and around the 2G12 epitope have been shown to interact with other potent, broadly neutralizing antibodies; therefore, this region constitutes a supersite of vulnerability on gp120. 24965454 8 102 theme 2G12 1611:1614 arg1 binding 1616:1622 2G12 binding 1611:1622 2G12 binding 1611:1622 These structures provide a basis for understanding 2G12 binding and neutralization, and our low-resolution model and glycan assignments provide a basis for higher-resolution studies to determine the molecular nature of the 2G12 epitope. 24965454 7 103 theme primary 1477:1483 arg1 binding 1494:1500 primary receptor binding 1477:1500 primary receptor binding 1477:1500 In addition, a reconstruction at ∼26 Å of the ternary complex formed by 2G12 Fab2, soluble CD4, and Env indicates that 2G12 may block membrane fusion by induced steric hindrance upon primary receptor binding, thereby abrogating Env's interaction with coreceptor(s). 24965454 6 104 theme crystal 1214:1220 arg1 structures 1222:1231 high-resolution crystal structures 1198:1231 high-resolution crystal structures 1198:1231 We also mapped relevant glycans in this epitope by fitting high-resolution crystal structures and by performing neutralization assays of glycan knockouts. 24965454 9 105 theme human 1819:1823 arg1 virus 1825:1829 a human virus 1817:1829 a human virus that results in the deaths of millions of people around the world each year 1817:1905 IMPORTANCE HIV-1 is a human virus that results in the deaths of millions of people around the world each year. 24965454 9 105 theme human 1819:1823 arg1 IMPORTANCE 1797:1806 IMPORTANCE HIV-1 1797:1812 IMPORTANCE HIV-1 1797:1812 IMPORTANCE HIV-1 is a human virus that results in the deaths of millions of people around the world each year. 24965454 5 106 with complex 994:1000 arg1 Env 1027:1029 a soluble, trimeric Env 1007:1029 a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 1007:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 24965454 0 107 with complex 26:32 arg1 Env 76:78 soluble and fully glycosylated HIV-1 Env 39:78 soluble and fully glycosylated HIV-1 Env 39:78 Structure of 2G12 Fab2 in complex with soluble and fully glycosylated HIV-1 Env by negative-stain single-particle electron microscopy. 24965454 2 108 theme conserved 369:377 arg1 glycans 388:394 conserved N-linked glycans 369:394 conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332 369:491 2G12 is a unique, domain-exchanged antibody that binds exclusively to conserved N-linked glycans that form the high-mannose patch on the gp120 outer domain centered on a glycan at position N332. 24965454 5 109 from reconstruction 963:976 arg1 complex 994:1000 complex 994:1000 complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope 994:1136 Here, we present a negative-stain single-particle electron microscopy reconstruction of 2G12 Fab2 in complex with a soluble, trimeric Env at ∼17-Å resolution that reveals the antibody's interaction with its native and fully glycosylated epitope. 26015571 8 0 from important 1501:1509 arg1 pathway 1527:1533 a biological pathway 1514:1533 a biological pathway 1514:1533 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 7 1 theme tight 1243:1247 arg1 integration 1249:1259 tight integration 1243:1259 tight integration 1243:1259 In addition, all three are up-regulated in TGF-β-induced EMT, suggesting tight integration within the EMT-signaling network. 26015571 1 2 theme biological 174:183 arg1 function 185:192 biological function 174:192 biological function 174:192 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 8 3 theme difficult-to-analyze 1416:1435 arg1 structures 1437:1446 difficult-to-analyze structures 1416:1446 difficult-to-analyze structures (e.g., proteoglycans, glycolipids) 1416:1481 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 8 3 theme difficult-to-analyze 1416:1435 arg1 glycolipids 1470:1480 glycolipids 1470:1480 glycolipids 1470:1480 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 4 4 theme promesenchymal 690:703 arg1 glycogenes 728:737 glycogenes 728:737 glycogenes 728:737 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 4 theme promesenchymal 690:703 arg1 enzymes 719:725 multiple promesenchymal glycosylation enzymes 681:725 multiple promesenchymal glycosylation enzymes (glycogenes) 681:738 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 5 5 theme -N-acetylgalactosaminide 920:943 arg1 enzymes 759:765 three enzymes 753:765 three enzymes 753:765 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 5 theme -N-acetylgalactosaminide 920:943 arg1 alpha-2,6-sialyltransferase 945:971 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 869:973 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5) 869:986 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 5 theme -N-acetylgalactosaminide 920:943 arg1 ST6GALNAC5 976:985 ST6GALNAC5 976:985 ST6GALNAC5 976:985 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 7 6 theme TGF-β-induced 1213:1225 arg1 EMT 1227:1229 TGF-β-induced EMT 1213:1229 TGF-β-induced EMT 1213:1229 In addition, all three are up-regulated in TGF-β-induced EMT, suggesting tight integration within the EMT-signaling network. 26015571 8 7 theme simple 1349:1354 arg1 miRNA 1319:1323 miRNA 1319:1323 miRNA 1319:1323 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 8 7 theme simple 1349:1354 arg1 proxy 1356:1360 a relatively simple proxy 1336:1360 a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway 1336:1533 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 0 8 theme proxy 6:10 arg1 approach 12:19 miRNA proxy approach 0:19 miRNA proxy approach 0:19 miRNA proxy approach reveals hidden functions of glycosylation. 26015571 3 9 theme regulatory 458:467 arg1 elements 469:476 regulatory elements 458:476 regulatory elements within a pathway using microRNA (miRNA) as a proxy 458:527 Herein we identify glycosylation enzymes acting as regulatory elements within a pathway using microRNA (miRNA) as a proxy. 26015571 0 10 theme miRNA 0:4 arg1 approach 12:19 miRNA proxy approach 0:19 miRNA proxy approach 0:19 miRNA proxy approach reveals hidden functions of glycosylation. 26015571 4 11 theme family 577:582 arg1 network 552:558 the target network 541:558 the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), 541:652 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 1 12 theme abundant 88:95 arg1 Glycosylation 64:76 Glycosylation 64:76 Glycosylation 64:76 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 1 12 theme abundant 88:95 arg1 modification 115:126 the most abundant posttranslational modification 79:126 the most abundant posttranslational modification 79:126 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 4 13 theme multiple 681:688 arg1 glycogenes 728:737 glycogenes 728:737 glycogenes 728:737 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 13 theme multiple 681:688 arg1 enzymes 719:725 multiple promesenchymal glycosylation enzymes 681:725 multiple promesenchymal glycosylation enzymes (glycogenes) 681:738 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 14 theme miRNA-200 567:575 arg1 regulators 596:605 regulators 596:605 regulators of epithelial-to-mesenchymal transition (EMT) 596:651 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 14 theme miRNA-200 567:575 arg1 family 577:582 the miRNA-200 family 563:582 the miRNA-200 family (miR-200f) 563:593 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 14 theme miRNA-200 567:575 arg1 miR-200f 585:592 miR-200f 585:592 miR-200f 585:592 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 1 15 theme posttranslational 97:113 arg1 Glycosylation 64:76 Glycosylation 64:76 Glycosylation 64:76 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 1 15 theme posttranslational 97:113 arg1 modification 115:126 the most abundant posttranslational modification 79:126 the most abundant posttranslational modification 79:126 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 8 16 theme biological 1516:1525 arg1 pathway 1527:1533 a biological pathway 1514:1533 a biological pathway 1514:1533 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 2 17 theme specific 318:325 arg1 glycans 327:333 the specific glycans 314:333 the specific glycans 314:333 Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. 26015571 8 18 dep proteoglycans 1455:1467 arg1 e.g. 1449:1452 e.g. 1449:1452 e.g. 1449:1452 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 8 19 theme rapid 1562:1566 arg1 identification 1568:1581 the rapid identification 1558:1581 the rapid identification of glycosylation enzymes driving disease states 1558:1629 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 5 20 theme traditional 1039:1049 arg1 methods 1051:1057 traditional methods 1039:1057 traditional methods 1039:1057 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 7 21 theme EMT-signaling 1272:1284 arg1 network 1286:1292 the EMT-signaling network 1268:1292 the EMT-signaling network 1268:1292 In addition, all three are up-regulated in TGF-β-induced EMT, suggesting tight integration within the EMT-signaling network. 26015571 8 22 from pathway 1527:1533 arg1 important 1501:1509 important 1501:1509 important 1501:1509 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 4 23 dep pinpoint 657:664 arg1 Leveraging 530:539 Leveraging 530:539 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), 530:652 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 5 24 theme alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3 870:918 arg1 enzymes 759:765 three enzymes 753:765 three enzymes 753:765 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 24 theme alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3 870:918 arg1 alpha-2,6-sialyltransferase 945:971 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 869:973 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5) 869:986 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 24 theme alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3 870:918 arg1 ST6GALNAC5 976:985 ST6GALNAC5 976:985 ST6GALNAC5 976:985 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 0 25 theme hidden 29:34 arg1 functions 36:44 hidden functions 29:44 hidden functions of glycosylation 29:61 miRNA proxy approach reveals hidden functions of glycosylation. 26015571 5 26 theme beta-galactoside 807:822 arg1 alpha-2,3-sialyltransferase 824:850 beta-galactoside alpha-2,3-sialyltransferase 5 807:852 beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5) 807:862 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 26 theme beta-galactoside 807:822 arg1 ST3GAL5 855:861 ST3GAL5 855:861 ST3GAL5 855:861 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 5 26 theme beta-galactoside 807:822 arg1 enzymes 759:765 three enzymes 753:765 three enzymes 753:765 We focus on three enzymes, beta-1,3-glucosyltransferase (B3GLCT), beta-galactoside alpha-2,3-sialyltransferase 5 (ST3GAL5), and (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 5 (ST6GALNAC5), encoding glycans that are difficult to analyze by traditional methods. 26015571 6 27 theme mesenchymal-to-epithelial 1132:1156 arg1 transition 1158:1167 mesenchymal-to-epithelial transition 1132:1167 mesenchymal-to-epithelial transition 1132:1167 Silencing these glycogenes phenocopied the effect of miR-200f, inducing mesenchymal-to-epithelial transition. 26015571 0 28 theme glycosylation 49:61 arg1 functions 36:44 hidden functions 29:44 hidden functions of glycosylation 29:61 miRNA proxy approach reveals hidden functions of glycosylation. 26015571 4 29 theme target 545:550 arg1 network 552:558 the target network 541:558 the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), 541:652 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 2 30 theme biological 387:396 arg1 process 398:404 a biological process 385:404 a biological process 385:404 Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. 26015571 8 31 theme enzymes 1600:1606 arg1 identification 1568:1581 the rapid identification 1558:1581 the rapid identification of glycosylation enzymes driving disease states 1558:1629 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 2 32 theme biosynthetic 353:364 arg1 enzymes 366:372 corresponding biosynthetic enzymes 339:372 corresponding biosynthetic enzymes 339:372 Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. 26015571 8 33 theme glycosylation 1586:1598 arg1 enzymes 1600:1606 glycosylation enzymes 1586:1606 glycosylation enzymes driving disease states 1586:1629 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 2 34 theme corresponding 339:351 arg1 enzymes 366:372 corresponding biosynthetic enzymes 339:372 corresponding biosynthetic enzymes 339:372 Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. 26015571 4 35 theme transition 636:645 arg1 regulators 596:605 regulators 596:605 regulators of epithelial-to-mesenchymal transition (EMT) 596:651 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 35 theme transition 636:645 arg1 family 577:582 the miRNA-200 family 563:582 the miRNA-200 family (miR-200f) 563:593 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 8 36 theme disease 1616:1622 arg1 states 1624:1629 disease states 1616:1629 disease states 1616:1629 Our work indicates that miRNA can act as a relatively simple proxy to decrypt which glycogenes, including those encoding difficult-to-analyze structures (e.g., proteoglycans, glycolipids), are functionally important in a biological pathway, setting the stage for the rapid identification of glycosylation enzymes driving disease states. 26015571 1 37 theme unprecedented 138:150 arg1 capacity 152:159 an unprecedented capacity 135:159 an unprecedented capacity for altering biological function 135:192 Glycosylation, the most abundant posttranslational modification, holds an unprecedented capacity for altering biological function. 26015571 4 38 theme epithelial-to-mesenchymal 610:634 arg1 transition 636:645 epithelial-to-mesenchymal transition 610:645 epithelial-to-mesenchymal transition (EMT) 610:651 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 38 theme epithelial-to-mesenchymal 610:634 arg1 EMT 648:650 EMT 648:650 EMT 648:650 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 6 39 theme miR-200f 1113:1120 arg1 effect 1103:1108 the effect 1099:1108 the effect of miR-200f 1099:1120 Silencing these glycogenes phenocopied the effect of miR-200f, inducing mesenchymal-to-epithelial transition. 26015571 3 40 theme glycosylation 426:438 arg1 enzymes 440:446 glycosylation enzymes 426:446 glycosylation enzymes acting as regulatory elements within a pathway using microRNA (miRNA) as a proxy 426:527 Herein we identify glycosylation enzymes acting as regulatory elements within a pathway using microRNA (miRNA) as a proxy. 26015571 2 41 theme biological 254:263 arg1 systems 265:271 biological systems 254:271 biological systems 254:271 Our ability to harness glycosylation as a means to control biological systems is hampered by our inability to pinpoint the specific glycans and corresponding biosynthetic enzymes underlying a biological process. 26015571 4 42 theme glycosylation 705:717 arg1 glycogenes 728:737 glycogenes 728:737 glycogenes 728:737 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 26015571 4 42 theme glycosylation 705:717 arg1 enzymes 719:725 multiple promesenchymal glycosylation enzymes 681:725 multiple promesenchymal glycosylation enzymes (glycogenes) 681:738 Leveraging the target network of the miRNA-200 family (miR-200f), regulators of epithelial-to-mesenchymal transition (EMT), we pinpoint genes encoding multiple promesenchymal glycosylation enzymes (glycogenes). 27287011 11 0 from changes 1747:1753 arg1 glycosylation 1758:1770 glycosylation 1758:1770 glycosylation 1758:1770 It could help in assessing the changes in glycosylation, controlling batch-to-batch consistency, and establishing acceptance limits according to the glycosylation changes, ensuring safety and efficacy. 27287011 6 1 theme high-resolution 876:890 arg1 HILIC-HRMS 911:920 HILIC-HRMS 911:920 HILIC-HRMS 911:920 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 6 1 theme high-resolution 876:890 arg1 spectrometer 897:908 a high-resolution mass spectrometer 874:908 a high-resolution mass spectrometer (HILIC-HRMS) 874:921 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 0 2 theme glycoproteins 183:195 arg1 profiles 159:166 N-glycosylation profiles 143:166 N-glycosylation profiles of therapeutic glycoproteins 143:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 6 3 theme interaction 829:839 arg1 chromatography 848:861 hydrophilic interaction liquid chromatography 817:861 hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS) 817:921 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 0 4 theme N-glycosylation 143:157 arg1 profiles 159:166 N-glycosylation profiles 143:166 N-glycosylation profiles of therapeutic glycoproteins 143:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 7 5 theme other 1059:1063 arg1 glycoproteins 1065:1077 other glycoproteins 1059:1077 other glycoproteins (such as ribonuclease B and fetuin) 1059:1113 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 0 6 theme therapeutic 171:181 arg1 glycoproteins 183:195 therapeutic glycoproteins 171:195 therapeutic glycoproteins 171:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 9 7 theme current 1502:1508 arg1 technique 1510:1518 current technique 1502:1518 current technique to detect N-glycosylation changes in batch-to-batch analysis 1502:1579 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 10 8 theme processing 1693:1702 arg1 approaches 1704:1713 MS and appropriate data processing approaches 1669:1713 MS and appropriate data processing approaches 1669:1713 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 0 9 from profiles 159:166 arg1 detection 119:127 a better detection 110:127 a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins 110:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 12 10 theme LC-MS-PCA 1976:1984 arg1 approach 1986:1993 LC-MS-PCA approach 1976:1993 LC-MS-PCA approach 1976:1993 Graphical abstract N-glycosylation characterization using LC-MS-PCA approach. 27287011 7 11 theme ribonuclease 1088:1099 arg1 B 1101:1101 ribonuclease B 1088:1101 ribonuclease B 1088:1101 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 8 12 theme glycosylation 1300:1312 arg1 changes 1314:1320 minor glycosylation changes 1294:1320 minor glycosylation changes 1294:1320 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 0 13 theme class 92:96 arg1 analogy 98:104 class analogy 92:104 class analogy 92:104 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 8 14 theme minor 1294:1298 arg1 changes 1314:1320 minor glycosylation changes 1294:1320 minor glycosylation changes 1294:1320 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 6 15 dep reduced 786:792 arg1 ii 782:783 ii 782:783 ii 782:783 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 10 16 theme HILIC-MS-PCA/SIMCA 1586:1603 arg1 approach 1605:1612 The HILIC-MS-PCA/SIMCA approach 1582:1612 The HILIC-MS-PCA/SIMCA approach 1582:1612 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 10 16 theme HILIC-MS-PCA/SIMCA 1586:1603 arg1 approach 1634:1641 highly sensitive approach 1617:1641 highly sensitive approach 1617:1641 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 10 17 theme data 1688:1691 arg1 processing 1693:1702 appropriate data processing 1676:1702 appropriate data processing 1676:1702 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 12 18 theme Graphical 1918:1926 arg1 characterization 1953:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization using LC-MS-PCA approach. 27287011 5 19 theme analytical 589:598 arg1 methods 620:626 analytical and data processing methods 589:626 methods 620:626 In this study, analytical and data processing methods were developed ensuring an easier characterization of glycoprofiles. 27287011 11 20 dep changes 1879:1885 arg1 ensuring 1888:1895 ensuring 1888:1895 ensuring safety and efficacy 1888:1915 It could help in assessing the changes in glycosylation, controlling batch-to-batch consistency, and establishing acceptance limits according to the glycosylation changes, ensuring safety and efficacy. 27287011 8 21 theme Principal 1160:1168 arg1 PCA 1190:1192 PCA 1190:1192 PCA 1190:1192 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 8 21 theme Principal 1160:1168 arg1 analysis 1180:1187 Principal component analysis 1160:1187 Principal component analysis (PCA) 1160:1193 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 6 22 theme liquid 841:846 arg1 chromatography 848:861 hydrophilic interaction liquid chromatography 817:861 hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS) 817:921 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 5 23 theme data 604:607 arg1 methods 620:626 analytical and data processing methods 589:626 methods 620:626 In this study, analytical and data processing methods were developed ensuring an easier characterization of glycoprofiles. 27287011 9 24 theme N-glycosylation 1530:1544 arg1 changes 1546:1552 N-glycosylation changes 1530:1552 N-glycosylation changes 1530:1552 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 11 25 theme batch-to-batch 1785:1798 arg1 consistency 1800:1810 batch-to-batch consistency 1785:1810 batch-to-batch consistency 1785:1810 It could help in assessing the changes in glycosylation, controlling batch-to-batch consistency, and establishing acceptance limits according to the glycosylation changes, ensuring safety and efficacy. 27287011 0 26 gly glycoproteins 183:195 arg1 glycoproteins 183:195 therapeutic glycoproteins 171:195 therapeutic glycoproteins 171:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 3 27 from changes 388:394 arg1 glycosylation 399:411 glycosylation 399:411 glycosylation 399:411 Nowadays, it is well accepted that changes in glycosylation may affect the safety and the efficacy of the therapeutic proteins. 27287011 5 28 theme processing 609:618 arg1 methods 620:626 analytical and data processing methods 589:626 methods 620:626 In this study, analytical and data processing methods were developed ensuring an easier characterization of glycoprofiles. 27287011 7 29 dep modified 1040:1047 arg1 digesting 1121:1129 digesting 1121:1129 digesting with enzyme (neuraminidase) 1121:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 7 29 dep modified 1040:1047 arg1 adding 1052:1057 adding 1052:1057 adding other glycoproteins (such as ribonuclease B and fetuin) 1052:1113 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 2 30 theme therapeutic 314:324 arg1 proteins 326:333 the approved therapeutic proteins 301:333 the approved therapeutic proteins 301:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 11 31 theme glycosylation 1865:1877 arg1 changes 1879:1885 the glycosylation changes 1861:1885 the glycosylation changes 1861:1885 It could help in assessing the changes in glycosylation, controlling batch-to-batch consistency, and establishing acceptance limits according to the glycosylation changes, ensuring safety and efficacy. 27287011 8 32 theme soft 1222:1225 arg1 modelling 1239:1247 soft independent modelling 1222:1247 soft independent modelling by class analogy (SIMCA) 1222:1272 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 0 33 theme LC-MS 0:4 arg1 analysis 6:13 LC-MS analysis 0:13 LC-MS analysis 0:13 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 12 34 theme abstract 1928:1935 arg1 characterization 1953:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization using LC-MS-PCA approach. 27287011 2 35 theme approved 305:312 arg1 proteins 326:333 the approved therapeutic proteins 301:333 the approved therapeutic proteins 301:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 8 36 theme independent 1227:1237 arg1 modelling 1239:1247 soft independent modelling 1222:1247 soft independent modelling by class analogy (SIMCA) 1222:1272 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 1 37 theme Therapeutic 198:208 arg1 proteins 210:217 Therapeutic proteins 198:217 Therapeutic proteins 198:217 Therapeutic proteins are among the top selling drugs in the pharmaceutical industry. 27287011 8 38 theme component 1170:1178 arg1 PCA 1190:1192 PCA 1190:1192 PCA 1190:1192 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 8 38 theme component 1170:1178 arg1 analysis 1180:1187 Principal component analysis 1160:1187 Principal component analysis (PCA) 1160:1193 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 9 39 from changes 1390:1396 arg1 N-glycosylation 1401:1415 N-glycosylation 1401:1415 N-glycosylation 1401:1415 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 6 40 theme PNGase 770:775 arg1 F 767:767 peptide-N-glycosidase F 745:767 peptide-N-glycosidase F (PNGase F) 745:778 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 6 40 theme PNGase 770:775 arg1 F 777:777 PNGase F 770:777 PNGase F 770:777 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 9 41 theme batch-to-batch 1557:1570 arg1 analysis 1572:1579 batch-to-batch analysis 1557:1579 batch-to-batch analysis 1557:1579 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 10 42 theme appropriate 1676:1686 arg1 processing 1693:1702 appropriate data processing 1676:1702 appropriate data processing 1676:1702 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 7 43 theme enzyme 1136:1141 arg1 neuraminidase 1144:1156 enzyme (neuraminidase) 1136:1157 enzyme (neuraminidase) 1136:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 0 44 theme better 112:117 arg1 detection 119:127 a better detection 110:127 a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins 110:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 6 45 theme hydrophilic 817:827 arg1 chromatography 848:861 hydrophilic interaction liquid chromatography 817:861 hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS) 817:921 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 4 46 theme glycan 555:560 arg1 structures 562:571 the glycan structures 551:571 the glycan structures 551:571 For this reason, it is important to characterize both the protein and the glycan structures. 27287011 3 47 theme therapeutic 459:469 arg1 proteins 471:478 the therapeutic proteins 455:478 the therapeutic proteins 455:478 Nowadays, it is well accepted that changes in glycosylation may affect the safety and the efficacy of the therapeutic proteins. 27287011 0 48 theme component 39:47 arg1 analysis 49:56 principal component analysis 29:56 principal component analysis 29:56 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 0 49 theme changes 132:138 arg1 detection 119:127 a better detection 110:127 a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins 110:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 7 50 theme Glycosylation 924:936 arg1 changes 938:944 Glycosylation changes 924:944 Glycosylation changes 924:944 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 7 51 gly glycoproteins 1065:1077 arg1 glycoproteins 1065:1077 other glycoproteins 1059:1077 other glycoproteins (such as ribonuclease B and fetuin) 1059:1113 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 8 52 theme class 1252:1256 arg1 SIMCA 1267:1271 SIMCA 1267:1271 SIMCA 1267:1271 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 8 52 theme class 1252:1256 arg1 analogy 1258:1264 class analogy 1252:1264 class analogy (SIMCA) 1252:1272 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 0 53 theme principal 29:37 arg1 analysis 49:56 principal component analysis 29:56 principal component analysis 29:56 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 6 54 dep analyzed 805:812 arg1 iii 800:802 iii 800:802 iii 800:802 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 1 55 theme pharmaceutical 258:271 arg1 industry 273:280 the pharmaceutical industry 254:280 the pharmaceutical industry 254:280 Therapeutic proteins are among the top selling drugs in the pharmaceutical industry. 27287011 9 56 theme HILIC-MS-PCA/SIMCA 1329:1346 arg1 approach 1348:1355 HILIC-MS-PCA/SIMCA approach 1329:1355 HILIC-MS-PCA/SIMCA approach 1329:1355 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 2 57 gly glycosylated 339:350 arg1 proteins 326:333 the approved therapeutic proteins 301:333 the approved therapeutic proteins 301:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 2 57 gly glycosylated 339:350 arg1 % 296:296 More than 60 % 283:296 More than 60 % of the approved therapeutic proteins 283:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 7 58 theme immunoglobulin 976:989 arg1 samples 993:999 human plasma immunoglobulin G samples 963:999 human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase) 963:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 11 59 theme acceptance 1830:1839 arg1 limits 1841:1846 acceptance limits 1830:1846 acceptance limits according to the glycosylation changes, ensuring safety and efficacy 1830:1915 It could help in assessing the changes in glycosylation, controlling batch-to-batch consistency, and establishing acceptance limits according to the glycosylation changes, ensuring safety and efficacy. 27287011 10 60 theme MS 1669:1670 arg1 approaches 1704:1713 MS and appropriate data processing approaches 1669:1713 MS and appropriate data processing approaches 1669:1713 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 6 61 theme peptide-N-glycosidase 745:765 arg1 F 767:767 peptide-N-glycosidase F 745:767 peptide-N-glycosidase F (PNGase F) 745:778 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 6 61 theme peptide-N-glycosidase 745:765 arg1 F 777:777 PNGase F 770:777 PNGase F 770:777 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 10 62 theme approaches 1704:1713 arg1 sensitivity 1654:1664 the sensitivity 1650:1664 the sensitivity of MS and appropriate data processing approaches 1650:1713 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 5 63 theme easier 655:660 arg1 characterization 662:677 an easier characterization 652:677 an easier characterization of glycoprofiles 652:694 In this study, analytical and data processing methods were developed ensuring an easier characterization of glycoprofiles. 27287011 7 64 theme G 991:991 arg1 samples 993:999 human plasma immunoglobulin G samples 963:999 human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase) 963:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 3 65 theme proteins 471:478 arg1 safety 428:433 the safety 424:433 the safety 424:433 Nowadays, it is well accepted that changes in glycosylation may affect the safety and the efficacy of the therapeutic proteins. 27287011 3 65 theme proteins 471:478 arg1 efficacy 443:450 the efficacy 439:450 the efficacy 439:450 Nowadays, it is well accepted that changes in glycosylation may affect the safety and the efficacy of the therapeutic proteins. 27287011 0 66 from changes 132:138 arg1 profiles 159:166 N-glycosylation profiles 143:166 N-glycosylation profiles of therapeutic glycoproteins 143:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 5 67 theme glycoprofiles 682:694 arg1 characterization 662:677 an easier characterization 652:677 an easier characterization of glycoprofiles 652:694 In this study, analytical and data processing methods were developed ensuring an easier characterization of glycoprofiles. 27287011 9 68 theme small 1384:1388 arg1 changes 1390:1396 small changes 1384:1396 small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms 1384:1490 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 7 69 theme human 963:967 arg1 samples 993:999 human plasma immunoglobulin G samples 963:999 human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase) 963:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 2 70 theme proteins 326:333 arg1 proteins 326:333 the approved therapeutic proteins 301:333 the approved therapeutic proteins 301:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 2 70 theme proteins 326:333 arg1 % 296:296 More than 60 % 283:296 More than 60 % of the approved therapeutic proteins 283:333 More than 60 % of the approved therapeutic proteins are glycosylated. 27287011 6 71 dep released 730:737 arg1 i 713:713 i 713:713 i 713:713 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 9 72 theme extracted-ion 1464:1476 arg1 chromatograms 1478:1490 the extracted-ion chromatograms 1460:1490 the extracted-ion chromatograms 1460:1490 Using HILIC-MS-PCA/SIMCA approach, it was possible to detect small changes in N-glycosylation, which had not been detected directly from the extracted-ion chromatograms, which is current technique to detect N-glycosylation changes in batch-to-batch analysis. 27287011 0 73 theme independent 67:77 arg1 modelling 79:87 soft independent modelling 62:87 soft independent modelling 62:87 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 7 74 dep glycoproteins 1065:1077 arg1 such 1080:1083 such 1080:1083 such 1080:1083 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 7 75 theme plasma 969:974 arg1 samples 993:999 human plasma immunoglobulin G samples 963:999 human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase) 963:1157 Glycosylation changes were analyzed in human plasma immunoglobulin G samples which had previously been artificially modified by adding other glycoproteins (such as ribonuclease B and fetuin) or by digesting with enzyme (neuraminidase). 27287011 0 76 from detection 119:127 arg1 profiles 159:166 N-glycosylation profiles 143:166 N-glycosylation profiles of therapeutic glycoproteins 143:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 0 77 theme soft 62:65 arg1 modelling 79:87 soft independent modelling 62:87 soft independent modelling 62:87 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 6 78 theme mass 892:895 arg1 HILIC-HRMS 911:920 HILIC-HRMS 911:920 HILIC-HRMS 911:920 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 6 78 theme mass 892:895 arg1 spectrometer 897:908 a high-resolution mass spectrometer 874:908 a high-resolution mass spectrometer (HILIC-HRMS) 874:921 N-glycans were (i) enzymatically released using peptide-N-glycosidase F (PNGase F), (ii) reduced, and (iii) analyzed by hydrophilic interaction liquid chromatography coupled to a high-resolution mass spectrometer (HILIC-HRMS). 27287011 8 79 used used 1279:1282 arg2 PCA 1190:1192 PCA 1190:1192 PCA 1190:1192 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 8 79 used used 1279:1282 arg2 analysis 1180:1187 Principal component analysis 1160:1187 Principal component analysis (PCA) 1160:1193 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 8 79 used used 1279:1282 arg2 classification 1199:1212 classification 1199:1212 classification through soft independent modelling by class analogy (SIMCA) 1199:1272 Principal component analysis (PCA) and classification through soft independent modelling by class analogy (SIMCA) were used to detect minor glycosylation changes. 27287011 12 80 theme N-glycosylation 1937:1951 arg1 characterization 1953:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization 1918:1968 Graphical abstract N-glycosylation characterization using LC-MS-PCA approach. 27287011 0 81 gly N-glycosylation 143:157 arg1 glycoproteins 183:195 therapeutic glycoproteins 171:195 therapeutic glycoproteins 171:195 LC-MS analysis combined with principal component analysis and soft independent modelling by class analogy for a better detection of changes in N-glycosylation profiles of therapeutic glycoproteins. 27287011 10 82 theme sensitive 1624:1632 arg1 approach 1605:1612 The HILIC-MS-PCA/SIMCA approach 1582:1612 The HILIC-MS-PCA/SIMCA approach 1582:1612 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 27287011 10 82 theme sensitive 1624:1632 arg1 approach 1634:1641 highly sensitive approach 1617:1641 highly sensitive approach 1617:1641 The HILIC-MS-PCA/SIMCA approach is highly sensitive approach due to the sensitivity of MS and appropriate data processing approaches. 26476458 5 0 theme glycan 682:687 arg1 structures 689:698 glycan structures 682:698 glycan structures 682:698 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 6 1 theme developers 828:837 arg1 team 820:823 an international team 803:823 an international team of developers and glycobiologists 803:857 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 3 2 theme nucleotides 416:426 arg1 sequences 388:396 linear sequences 381:396 linear sequences of amino acids or nucleotides 381:426 The structure of glycans, however, differs greatly from DNA and proteins in that they are branched, as opposed to linear sequences of amino acids or nucleotides. 26476458 6 3 from http 942:945 arg1 available 929:937 available 929:937 available 929:937 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 4 4 theme let 488:490 arg1 curation 504:511 let alone their curation 488:511 let alone their curation 488:511 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 4 4 theme let 488:490 arg1 storage 444:450 the storage 440:450 the storage of glycan information in databases 440:485 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 6 5 theme glycan 1014:1019 arg1 structures 1021:1030 glycan structures 1014:1030 glycan structures 1014:1030 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 5 6 theme many 560:563 arg1 efforts 576:582 many duplicated efforts 560:582 many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary 560:791 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 6 7 theme accession 1078:1086 arg1 numbers 1088:1094 accession numbers 1078:1094 accession numbers for each of these registered entries 1078:1131 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 6 8 theme international 806:818 arg1 team 820:823 an international team 803:823 an international team of developers and glycobiologists 803:857 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 5 9 theme duplicated 565:574 arg1 efforts 576:582 many duplicated efforts 560:582 many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary 560:791 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 2 10 theme other 226:230 arg1 biomolecules 232:243 other biomolecules 226:243 other biomolecules 226:243 They cover the surfaces of many cells, serving as the 'face' of cells, whereby other biomolecules and viruses interact. 26476458 3 11 theme acids 407:411 arg1 sequences 388:396 linear sequences 381:396 linear sequences of amino acids or nucleotides 381:426 The structure of glycans, however, differs greatly from DNA and proteins in that they are branched, as opposed to linear sequences of amino acids or nucleotides. 26476458 5 12 theme identified 754:763 arg1 structure 772:780 every identified glycan structure 748:780 every identified glycan structure 748:780 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 7 13 theme chemical 1244:1251 arg1 burden 1281:1286 a burden 1279:1286 a burden to integrate glycomics databases in the past 1279:1331 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 26476458 7 13 theme chemical 1244:1251 arg1 structure 1253:1261 chemical structure 1244:1261 chemical structure 1244:1261 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 26476458 5 14 theme international 653:665 arg1 repository 667:676 an international repository 650:676 an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, 650:781 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 0 15 theme 1.0--The 10:17 arg1 structure 40:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure repository. 26476458 1 16 theme third 87:91 arg1 class 99:103 the third major class 83:103 the third major class of biopolymers, next to DNA and proteins 83:144 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 1 16 theme third 87:91 arg1 Glycans 62:68 Glycans 62:68 Glycans 62:68 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 6 17 theme centralized 978:988 arg1 resource 990:997 a centralized resource 976:997 a centralized resource for depositing glycan structures, compositions and topologies 976:1059 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 0 18 theme GlyTouCan 0:8 arg1 structure 40:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure repository. 26476458 1 19 theme major 93:97 arg1 class 99:103 the third major class 83:103 the third major class of biopolymers, next to DNA and proteins 83:144 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 1 19 theme major 93:97 arg1 Glycans 62:68 Glycans 62:68 Glycans 62:68 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 0 20 theme glycan 33:38 arg1 structure 40:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure repository. 26476458 4 21 theme information 462:472 arg1 curation 504:511 let alone their curation 488:511 let alone their curation 488:511 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 4 21 theme information 462:472 arg1 storage 444:450 the storage 440:450 the storage of glycan information in databases 440:485 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 4 21 theme information 462:472 arg1 problem 535:541 a difficult problem 523:541 a difficult problem 523:541 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 5 22 theme unique 707:712 arg1 numbers 724:730 unique accession numbers 707:730 unique accession numbers 707:730 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 3 23 theme linear 381:386 arg1 sequences 388:396 linear sequences 381:396 linear sequences of amino acids or nucleotides 381:426 The structure of glycans, however, differs greatly from DNA and proteins in that they are branched, as opposed to linear sequences of amino acids or nucleotides. 26476458 0 24 theme international 19:31 arg1 structure 40:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure repository. 26476458 4 25 theme difficult 525:533 arg1 problem 535:541 a difficult problem 523:541 a difficult problem 523:541 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 4 25 theme difficult 525:533 arg1 storage 444:450 the storage 440:450 the storage of glycan information in databases 440:485 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 5 26 theme different 622:630 arg1 databases 632:640 different databases 622:640 different databases 622:640 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 5 27 theme glycan 765:770 arg1 structure 772:780 every identified glycan structure 748:780 every identified glycan structure 748:780 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 4 28 from storage 444:450 arg1 databases 477:485 databases 477:485 databases 477:485 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 0 29 dep repository 50:59 arg1 structure 40:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure 0:48 GlyTouCan 1.0--The international glycan structure repository. 26476458 1 30 theme biopolymers 108:118 arg1 class 99:103 the third major class 83:103 the third major class of biopolymers, next to DNA and proteins 83:144 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 1 30 theme biopolymers 108:118 arg1 Glycans 62:68 Glycans 62:68 Glycans 62:68 Glycans are known as the third major class of biopolymers, next to DNA and proteins. 26476458 5 31 theme accession 714:722 arg1 numbers 724:730 unique accession numbers 707:730 unique accession numbers 707:730 This has caused many duplicated efforts when integration is attempted between different databases, making an international repository for glycan structures, where unique accession numbers are assigned to every identified glycan structure, necessary. 26476458 4 32 theme glycan 455:460 arg1 information 462:472 glycan information 455:472 glycan information 455:472 Therefore, the storage of glycan information in databases, let alone their curation, has been a difficult problem. 26476458 7 33 theme accession 1209:1217 arg1 number 1219:1224 accession number 1209:1224 accession number 1209:1224 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 26476458 6 34 theme glycobiologists 843:857 arg1 team 820:823 an international team 803:823 an international team of developers and glycobiologists 803:857 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 2 35 theme cells 179:183 arg1 surfaces 162:169 the surfaces 158:169 the surfaces 158:169 They cover the surfaces of many cells, serving as the 'face' of cells, whereby other biomolecules and viruses interact. 26476458 2 36 theme cells 211:215 arg1 face 202:205 the 'face' 197:206 the 'face' of cells 197:215 They cover the surfaces of many cells, serving as the 'face' of cells, whereby other biomolecules and viruses interact. 26476458 7 37 theme glycomics 1301:1309 arg1 databases 1311:1319 glycomics databases 1301:1319 glycomics databases 1301:1319 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 26476458 2 38 theme many 174:177 arg1 cells 179:183 many cells 174:183 many cells 174:183 They cover the surfaces of many cells, serving as the 'face' of cells, whereby other biomolecules and viruses interact. 26476458 3 39 theme amino 401:405 arg1 acids 407:411 amino acids 401:411 amino acids 401:411 The structure of glycans, however, differs greatly from DNA and proteins in that they are branched, as opposed to linear sequences of amino acids or nucleotides. 26476458 6 40 dep repository 893:902 arg1 called 905:910 called 905:910 called GlyTouCan 905:920 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 6 40 dep repository 893:902 arg1 available 929:937 available 929:937 available 929:937 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 6 41 theme registered 1114:1123 arg1 entries 1125:1131 these registered entries 1108:1131 these registered entries 1108:1131 As such, an international team of developers and glycobiologists have collaborated to develop this repository, called GlyTouCan and is available at http://glytoucan.org/, to provide a centralized resource for depositing glycan structures, compositions and topologies, and to retrieve accession numbers for each of these registered entries. 26476458 7 42 theme reference 1171:1179 arg1 structures 1188:1197 reference glycan structures 1171:1197 reference glycan structures 1171:1197 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 26476458 3 43 theme glycans 284:290 arg1 structure 271:279 The structure 267:279 The structure of glycans 267:290 The structure of glycans, however, differs greatly from DNA and proteins in that they are branched, as opposed to linear sequences of amino acids or nucleotides. 26476458 7 44 theme glycan 1181:1186 arg1 structures 1188:1197 reference glycan structures 1171:1197 reference glycan structures 1171:1197 This will thus enable researchers to reference glycan structures simply by accession number, as opposed to by chemical structure, which has been a burden to integrate glycomics databases in the past. 27089023 4 0 from glycopeptides 893:905 arg1 differentiation 822:836 differentiation 822:836 differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides 822:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 1 theme O-linked 884:891 arg1 glycopeptides 893:905 O-linked glycopeptides 884:905 O-linked glycopeptides 884:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 2 2 with approaches 227:236 arg1 spectrometry 248:259 mass spectrometry 243:259 mass spectrometry 243:259 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 5 3 from elucidation 994:1004 arg1 level 1042:1046 a site-specific level 1026:1046 a site-specific level 1026:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 3 4 with level 595:599 arg1 analyses 616:623 proteomic analyses 606:623 proteomic analyses 606:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 3 5 theme glycan 546:551 arg1 information 564:574 glycan structural information 546:574 glycan structural information on a site-specific level with proteomic analyses 546:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 5 6 theme structural 983:992 arg1 elucidation 994:1004 rapid and complete structural elucidation 964:1004 rapid and complete structural elucidation of glycan chains at a site-specific level 964:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 5 7 theme rapid 964:968 arg1 elucidation 994:1004 rapid and complete structural elucidation 964:1004 rapid and complete structural elucidation of glycan chains at a site-specific level 964:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 3 8 theme structural 553:562 arg1 information 564:574 glycan structural information 546:574 glycan structural information on a site-specific level with proteomic analyses 546:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 4 9 theme glycoproteomics 710:724 arg1 platforms 726:734 conventional glycoproteomics platforms 697:734 conventional glycoproteomics platforms 697:734 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 5 10 theme glycan 1009:1014 arg1 chains 1016:1021 glycan chains 1009:1021 glycan chains at a site-specific level 1009:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 5 11 from level 1042:1046 arg1 elucidation 994:1004 rapid and complete structural elucidation 964:1004 rapid and complete structural elucidation of glycan chains at a site-specific level 964:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 5 11 from level 1042:1046 arg1 chains 1016:1021 glycan chains 1009:1021 glycan chains at a site-specific level 1009:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 1 12 theme several 177:183 arg1 techniques 185:194 several techniques 177:194 several techniques 177:194 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 3 13 theme mobility 420:427 arg1 IMS 443:445 IMS 443:445 IMS 443:445 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 3 13 theme mobility 420:427 arg1 spectrometry 429:440 ion mobility spectrometry 416:440 ion mobility spectrometry (IMS) 416:446 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 5 14 theme chains 1016:1021 arg1 elucidation 994:1004 rapid and complete structural elucidation 964:1004 rapid and complete structural elucidation of glycan chains at a site-specific level 964:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 4 15 gly glycopeptides 893:905 arg2 glycopeptides 893:905 O-linked glycopeptides 884:905 O-linked glycopeptides 884:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 1 16 theme Detailed 83:90 arg1 process 153:159 a tedious process 143:159 a tedious process often involving several techniques 143:194 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 1 16 theme Detailed 83:90 arg1 elucidation 103:113 Detailed structural elucidation 83:113 Detailed structural elucidation of protein glycosylation 83:138 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 4 17 from N- 877:878 arg1 differentiation 822:836 differentiation 822:836 differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides 822:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 0 18 theme Site-Specific 0:12 arg1 Mapping 14:20 Site-Specific Mapping 0:20 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 0:81 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 1 19 theme structural 92:101 arg1 process 153:159 a tedious process 143:159 a tedious process often involving several techniques 143:194 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 1 19 theme structural 92:101 arg1 elucidation 103:113 Detailed structural elucidation 83:113 Detailed structural elucidation of protein glycosylation 83:138 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 0 20 theme Sialic 25:30 arg1 Isomers 45:51 Sialic Acid Linkage Isomers 25:51 Sialic Acid Linkage Isomers 25:51 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 4 21 theme additional 741:750 arg1 information 752:762 additional information 741:762 additional information regarding glycan structure 741:789 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 22 theme linkage 853:859 arg1 isomers 861:867 sialic acid linkage isomers 841:867 sialic acid linkage isomers on both N- and O-linked glycopeptides 841:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 23 theme glycan 774:779 arg1 structure 781:789 glycan structure 774:789 glycan structure 774:789 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 24 theme acid 848:851 arg1 isomers 861:867 sialic acid linkage isomers 841:867 sialic acid linkage isomers on both N- and O-linked glycopeptides 841:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 3 25 theme site-specific 581:593 arg1 level 595:599 a site-specific level 579:599 a site-specific level with proteomic analyses 579:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 2 26 theme Glycomics 197:205 arg1 approaches 227:236 Glycomics and glycoproteomics approaches 197:236 Glycomics and glycoproteomics approaches with mass spectrometry 197:259 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 4 27 theme IMS 690:692 arg1 addition 678:685 the addition 674:685 the addition of IMS to conventional glycoproteomics platforms 674:734 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 27 theme IMS 690:692 arg1 useful 811:816 useful 811:816 useful 811:816 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 1 28 theme tedious 145:151 arg1 process 153:159 a tedious process 143:159 a tedious process often involving several techniques 143:194 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 1 28 theme tedious 145:151 arg1 elucidation 103:113 Detailed structural elucidation 83:113 Detailed structural elucidation of protein glycosylation 83:138 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 0 29 theme Linkage 37:43 arg1 Isomers 45:51 Sialic Acid Linkage Isomers 25:51 Sialic Acid Linkage Isomers 25:51 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 4 30 theme conventional 697:708 arg1 platforms 726:734 conventional glycoproteomics platforms 697:734 conventional glycoproteomics platforms 697:734 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 31 link O-linked 884:891 arg1 glycopeptides 893:905 O-linked glycopeptides 884:905 O-linked glycopeptides 884:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 3 32 theme proteomic 606:614 arg1 analyses 616:623 proteomic analyses 606:623 proteomic analyses 606:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 0 33 theme Acid 32:35 arg1 Isomers 45:51 Sialic Acid Linkage Isomers 25:51 Sialic Acid Linkage Isomers 25:51 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 4 34 theme sialic 841:846 arg1 isomers 861:867 sialic acid linkage isomers 841:867 sialic acid linkage isomers on both N- and O-linked glycopeptides 841:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 35 from isomers 861:867 arg1 N- 877:878 N- 877:878 N- 877:878 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 35 from isomers 861:867 arg1 glycopeptides 893:905 O-linked glycopeptides 884:905 O-linked glycopeptides 884:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 3 36 theme ion 416:418 arg1 IMS 443:445 IMS 443:445 IMS 443:445 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 3 36 theme ion 416:418 arg1 spectrometry 429:440 ion mobility spectrometry 416:440 ion mobility spectrometry (IMS) 416:446 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 5 37 theme further 913:919 arg1 development 921:931 further development 913:931 further development 913:931 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 4 38 theme isomers 861:867 arg1 differentiation 822:836 differentiation 822:836 differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides 822:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 1 39 theme protein 118:124 arg1 glycosylation 126:138 protein glycosylation 118:138 protein glycosylation 118:138 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 0 40 theme Isomers 45:51 arg1 Mapping 14:20 Site-Specific Mapping 0:20 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 0:81 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 2 41 theme mass 243:246 arg1 spectrometry 248:259 mass spectrometry 243:259 mass spectrometry 243:259 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 1 42 theme glycosylation 126:138 arg1 process 153:159 a tedious process 143:159 a tedious process often involving several techniques 143:194 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 1 42 theme glycosylation 126:138 arg1 elucidation 103:113 Detailed structural elucidation 83:113 Detailed structural elucidation of protein glycosylation 83:138 Detailed structural elucidation of protein glycosylation is a tedious process often involving several techniques. 27089023 3 43 from information 564:574 arg1 level 595:599 a site-specific level 579:599 a site-specific level with proteomic analyses 579:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 2 44 theme glycan 288:293 arg1 profiling 295:303 glycan profiling 288:303 glycan profiling 288:303 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 2 45 theme isobaric 349:356 arg1 species 358:364 isobaric species 349:364 isobaric species such as linkage and positional isomers 349:403 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 2 45 theme isobaric 349:356 arg1 linkage 374:380 linkage 374:380 linkage 374:380 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 2 45 theme isobaric 349:356 arg1 isomers 397:403 positional isomers 386:403 positional isomers 386:403 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 5 46 theme complete 974:981 arg1 elucidation 994:1004 rapid and complete structural elucidation 964:1004 rapid and complete structural elucidation of glycan chains at a site-specific level 964:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 2 47 theme glycoproteomics 211:225 arg1 approaches 227:236 Glycomics and glycoproteomics approaches 197:236 Glycomics and glycoproteomics approaches with mass spectrometry 197:259 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 3 48 theme isobaric 486:493 arg1 oligosaccharides 495:510 isobaric oligosaccharides 486:510 isobaric oligosaccharides 486:510 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 27089023 0 49 theme Mobility 60:67 arg1 Spectrometry 69:80 Ion Mobility Spectrometry 56:80 Ion Mobility Spectrometry 56:80 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 5 50 theme site-specific 1028:1040 arg1 level 1042:1046 a site-specific level 1026:1046 a site-specific level 1026:1046 With further development IMS may hold the potential for rapid and complete structural elucidation of glycan chains at a site-specific level. 27089023 4 51 from differentiation 822:836 arg1 N- 877:878 N- 877:878 N- 877:878 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 4 51 from differentiation 822:836 arg1 glycopeptides 893:905 O-linked glycopeptides 884:905 O-linked glycopeptides 884:905 Here, we report that the addition of IMS to conventional glycoproteomics platforms adds additional information regarding glycan structure and is particularly useful for differentiation of sialic acid linkage isomers on both N- and O-linked glycopeptides. 27089023 2 52 theme positional 386:395 arg1 isomers 397:403 positional isomers 386:403 positional isomers 386:403 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 0 53 theme Ion 56:58 arg1 Spectrometry 69:80 Ion Mobility Spectrometry 56:80 Ion Mobility Spectrometry 56:80 Site-Specific Mapping of Sialic Acid Linkage Isomers by Ion Mobility Spectrometry. 27089023 2 54 theme rapid 269:273 arg1 platform 275:282 a rapid platform 267:282 a rapid platform for glycan profiling 267:303 Glycomics and glycoproteomics approaches with mass spectrometry offer a rapid platform for glycan profiling but are limited by the inability to resolve isobaric species such as linkage and positional isomers. 27089023 3 55 theme IMS 532:534 arg1 utility 521:527 the utility 517:527 the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses 517:623 Recently, ion mobility spectrometry (IMS) has been shown to effectively resolve isobaric oligosaccharides, but the utility of IMS to obtain glycan structural information on a site-specific level with proteomic analyses has yet to be investigated. 26940433 9 0 theme downstream 1755:1764 arg1 signaling 1781:1789 downstream trans-synaptic signaling 1755:1789 downstream trans-synaptic signaling 1755:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 10 1 theme novel 2028:2032 arg1 strategies 2056:2065 novel therapeutic treatment strategies 2028:2065 novel therapeutic treatment strategies 2028:2065 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 10 2 theme Drosophila 1840:1849 arg1 model 1863:1867 this Drosophila CDG disease model 1835:1867 this Drosophila CDG disease model 1835:1867 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 9 3 theme signaling 1781:1789 arg1 Loss 1701:1704 Loss 1701:1704 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling 1701:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 10 4 theme new 1880:1882 arg1 avenue 1884:1889 a new avenue 1878:1889 a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments 1878:1981 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 5 5 theme PMM2 880:883 arg1 knockdown 858:866 RNAi-targeted knockdown 844:866 RNAi-targeted knockdown of neuronal PMM2 844:883 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 6 theme structural 1262:1271 arg1 architecture 1273:1284 structural architecture 1262:1284 structural architecture 1262:1284 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 4 7 theme first 697:701 arg1 model 721:725 the first Drosophila CDG-Ia model 693:725 the first Drosophila CDG-Ia model 693:725 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 1 8 theme growing 183:189 arg1 family 191:196 a rapidly growing family 173:196 a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins 173:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 5 9 theme pmm2-null 745:753 arg1 mutants 766:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 3 10 theme phosphomannomutase 594:611 arg1 type 613:616 phosphomannomutase type 2 594:618 phosphomannomutase type 2 (PMM2) mutations 594:635 The most common CDG, CDG-Ia (PMM2-CDG), arises from phosphomannomutase type 2 (PMM2) mutations. 26940433 3 10 theme phosphomannomutase 594:611 arg1 PMM2 621:624 PMM2 621:624 PMM2 621:624 The most common CDG, CDG-Ia (PMM2-CDG), arises from phosphomannomutase type 2 (PMM2) mutations. 26940433 5 11 theme strong 898:903 arg1 shift 905:909 a strong shift 896:909 a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality 896:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 1 12 theme Congenital 117:126 arg1 disorders 128:136 Congenital disorders 117:136 Congenital disorders of glycosylation (CDGs) 117:160 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 1 12 theme Congenital 117:126 arg1 CDGs 156:159 CDGs 156:159 CDGs 156:159 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 8 13 theme heparan 1582:1588 arg1 proteoglycan 1598:1609 heparan sulfate proteoglycan 1582:1609 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp) 1578:1653 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 13 theme heparan 1582:1588 arg1 HSPG 1612:1615 HSPG 1612:1615 HSPG 1612:1615 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 10 14 theme molecular 1926:1934 arg1 mechanisms 1936:1945 cellular and molecular mechanisms 1913:1945 cellular and molecular mechanisms underlying neurological impairments 1913:1981 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 5 15 from shift 905:909 arg1 abundance 918:926 the abundance 914:926 the abundance of pauci-mannose glycan, progressive incoordination and later lethality 914:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 1 16 theme diseases 207:214 arg1 family 191:196 a rapidly growing family 173:196 a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins 173:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 8 17 theme proteoglycan 1598:1609 arg1 Dlp 1650:1652 Dlp 1650:1652 Dlp 1650:1652 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 17 theme proteoglycan 1598:1609 arg1 protein 1641:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein 1578:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp) 1578:1653 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 1 18 from mutations 241:249 arg1 genes 254:258 genes 254:258 genes driving the production and modification of glycoproteins 254:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 10 19 theme cellular 1913:1920 arg1 mechanisms 1936:1945 cellular and molecular mechanisms 1913:1945 cellular and molecular mechanisms underlying neurological impairments 1913:1981 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 5 20 theme later 984:988 arg1 lethality 990:998 later lethality 984:998 later lethality 984:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 5 21 dep display 774:780 arg1 whereas 836:842 whereas 836:842 whereas 836:842 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 8 22 theme co-receptor 1618:1628 arg1 Dlp 1650:1652 Dlp 1650:1652 Dlp 1650:1652 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 22 theme co-receptor 1618:1628 arg1 protein 1641:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein 1578:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp) 1578:1653 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 4 23 theme CDG-Ia 714:719 arg1 model 721:725 the first Drosophila CDG-Ia model 693:725 the first Drosophila CDG-Ia model 693:725 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 5 24 theme early 819:823 arg1 lethality 825:833 early lethality 819:833 early lethality 819:833 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 0 25 theme congenital 65:74 arg1 disorder 76:83 a new Drosophila congenital disorder 48:83 a new Drosophila congenital disorder of glycosylation disease model 48:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 5 26 theme incoordination 965:978 arg1 abundance 918:926 the abundance 914:926 the abundance of pauci-mannose glycan, progressive incoordination and later lethality 914:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 9 27 theme synaptic 1709:1716 arg1 MMP2 1718:1721 synaptic MMP2 1709:1721 synaptic MMP2 1709:1721 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 8 28 theme trans-synaptic 1509:1522 arg1 signaling 1524:1532 trans-synaptic signaling 1509:1532 trans-synaptic signaling by the Wnt protein Wingless (Wg) 1509:1565 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 0 29 theme disease 102:108 arg1 model 110:114 glycosylation disease model 88:114 glycosylation disease model 88:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 5 30 theme lifespan 1056:1063 arg1 symptoms 1034:1041 human CDG-Ia symptoms 1021:1041 human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development 1021:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 31 theme junction 1181:1188 arg1 Analyses 1121:1128 Analyses 1121:1128 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) 1121:1194 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 0 32 theme Drosophila 54:63 arg1 disorder 76:83 a new Drosophila congenital disorder 48:83 a new Drosophila congenital disorder of glycosylation disease model 48:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 9 33 theme MMP2 1718:1721 arg1 Loss 1701:1704 Loss 1701:1704 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling 1701:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 0 34 theme Synaptic 0:7 arg1 roles 9:13 Synaptic roles 0:13 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 0:115 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 6 35 theme Drosophila 1156:1165 arg1 NMJ 1191:1193 NMJ 1191:1193 NMJ 1191:1193 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 6 35 theme Drosophila 1156:1165 arg1 junction 1181:1188 the well-characterized Drosophila neuromuscular junction 1133:1188 the well-characterized Drosophila neuromuscular junction (NMJ) 1133:1194 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 8 36 theme Wnt 1541:1543 arg1 Wg 1563:1564 Wg 1563:1564 Wg 1563:1564 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 36 theme Wnt 1541:1543 arg1 Wingless 1553:1560 the Wnt protein Wingless 1537:1560 the Wnt protein Wingless (Wg) 1537:1565 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 6 37 from defects 1246:1252 arg1 architecture 1273:1284 structural architecture 1262:1284 structural architecture 1262:1284 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 6 37 from defects 1246:1252 arg1 neurotransmission 1301:1317 functional neurotransmission 1290:1317 functional neurotransmission 1290:1317 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 5 38 theme CDG-Ia 1027:1032 arg1 symptoms 1034:1041 human CDG-Ia symptoms 1021:1041 human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development 1021:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 7 39 theme matrix 1461:1466 arg1 MMPs 1488:1491 MMPs 1488:1491 MMPs 1488:1491 NMJ synaptogenesis is driven by intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs). 26940433 7 39 theme matrix 1461:1466 arg1 metalloproteinases 1468:1485 matrix metalloproteinases 1461:1485 matrix metalloproteinases (MMPs) 1461:1492 NMJ synaptogenesis is driven by intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs). 26940433 8 40 theme MMP 1687:1689 arg1 activity 1691:1698 synaptic MMP activity 1678:1698 synaptic MMP activity 1678:1698 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 5 41 theme RNAi-targeted 844:856 arg1 knockdown 858:866 RNAi-targeted knockdown 844:866 RNAi-targeted knockdown of neuronal PMM2 844:883 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 1 42 theme glycoproteins 303:315 arg1 modification 287:298 modification 287:298 modification 287:298 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 1 42 theme glycoproteins 303:315 arg1 production 272:281 production 272:281 production 272:281 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 4 43 dep generation 658:667 arg1 the 654:656 the 654:656 the 654:656 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 0 44 theme new 50:52 arg1 disorder 76:83 a new Drosophila congenital disorder 48:83 a new Drosophila congenital disorder of glycosylation disease model 48:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 2 45 theme multisystemic 369:381 arg1 defects 383:389 multisystemic defects 369:389 multisystemic defects that include severe neurological impairments 369:434 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 9 46 theme ligand 1727:1732 arg1 Loss 1701:1704 Loss 1701:1704 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling 1701:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 5 47 theme neural 1101:1106 arg1 development 1108:1118 defective neural development 1091:1118 defective neural development 1091:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 5 48 theme movement 1066:1073 arg1 impairments 1075:1085 movement impairments 1066:1085 movement impairments 1066:1085 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 49 theme glycosylation 1212:1224 arg1 loss 1226:1229 synaptic glycosylation loss 1203:1229 synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission 1203:1317 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 2 50 theme severe 404:409 arg1 impairments 424:434 severe neurological impairments 404:434 severe neurological impairments 404:434 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 1 51 theme heritable 231:239 arg1 mutations 241:249 heritable mutations 231:249 heritable mutations in genes driving the production and modification of glycoproteins 231:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 2 52 from requirement 471:481 arg1 system 534:539 the nervous system 522:539 the nervous system 522:539 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 7 53 theme extracellular 1391:1403 arg1 synaptomatrix 1405:1417 an extracellular synaptomatrix 1388:1417 an extracellular synaptomatrix 1388:1417 NMJ synaptogenesis is driven by intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs). 26940433 10 54 theme therapeutic 2034:2044 arg1 strategies 2056:2065 novel therapeutic treatment strategies 2028:2065 novel therapeutic treatment strategies 2028:2065 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 10 55 theme CDG 1851:1853 arg1 model 1863:1867 this Drosophila CDG disease model 1835:1867 this Drosophila CDG disease model 1835:1867 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 9 56 theme trans-synaptic 1766:1779 arg1 signaling 1781:1789 downstream trans-synaptic signaling 1755:1789 downstream trans-synaptic signaling 1755:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 9 57 theme Wg 1724:1725 arg1 ligand 1727:1732 Wg ligand 1724:1732 Wg ligand 1724:1732 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 2 58 theme resulting 322:330 arg1 hypoglycosylation 344:360 The resulting symptomatic hypoglycosylation 318:360 The resulting symptomatic hypoglycosylation 318:360 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 6 59 theme functional 1290:1299 arg1 neurotransmission 1301:1317 functional neurotransmission 1290:1317 functional neurotransmission 1290:1317 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 9 60 theme PMM2 1803:1806 arg1 knockdown 1808:1816 PMM2 knockdown 1803:1816 PMM2 knockdown 1803:1816 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 3 61 theme type 613:616 arg1 mutations 627:635 phosphomannomutase type 2 (PMM2) mutations 594:635 phosphomannomutase type 2 (PMM2) mutations 594:635 The most common CDG, CDG-Ia (PMM2-CDG), arises from phosphomannomutase type 2 (PMM2) mutations. 26940433 0 62 from roles 9:13 arg1 disorder 76:83 a new Drosophila congenital disorder 48:83 a new Drosophila congenital disorder of glycosylation disease model 48:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 5 63 theme CRISPR-generated 728:743 arg1 mutants 766:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 8 64 theme sulfate 1590:1596 arg1 proteoglycan 1598:1609 heparan sulfate proteoglycan 1582:1609 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp) 1578:1653 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 64 theme sulfate 1590:1596 arg1 HSPG 1612:1615 HSPG 1612:1615 HSPG 1612:1615 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 1 65 theme glycosylation 141:153 arg1 disorders 128:136 Congenital disorders 117:136 Congenital disorders of glycosylation (CDGs) 117:160 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 1 65 theme glycosylation 141:153 arg1 CDGs 156:159 CDGs 156:159 CDGs 156:159 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 5 66 theme Drosophila 755:764 arg1 mutants 766:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants 728:772 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 10 67 theme mechanisms 1936:1945 arg1 dissection 1899:1908 the dissection 1895:1908 the dissection of cellular and molecular mechanisms underlying neurological impairments 1895:1981 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 5 68 theme pauci-mannose 931:943 arg1 glycan 945:950 pauci-mannose glycan 931:950 pauci-mannose glycan 931:950 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 1 69 theme human 201:205 arg1 diseases 207:214 human diseases 201:214 human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins 201:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 5 70 theme lethality 990:998 arg1 abundance 918:926 the abundance 914:926 the abundance of pauci-mannose glycan, progressive incoordination and later lethality 914:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 8 71 theme Dally-like 1630:1639 arg1 Dlp 1650:1652 Dlp 1650:1652 Dlp 1650:1652 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 71 theme Dally-like 1630:1639 arg1 protein 1641:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein 1578:1647 the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp) 1578:1653 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 7 72 theme intercellular 1352:1364 arg1 signals 1366:1372 intercellular signals 1352:1372 intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs) 1352:1492 NMJ synaptogenesis is driven by intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs). 26940433 10 73 theme neurological 1958:1969 arg1 impairments 1971:1981 neurological impairments 1958:1981 neurological impairments 1958:1981 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 1 74 gly glycoproteins 303:315 arg1 glycoproteins 303:315 glycoproteins 303:315 glycoproteins 303:315 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 5 75 theme glycan 945:950 arg1 abundance 918:926 the abundance 914:926 the abundance of pauci-mannose glycan, progressive incoordination and later lethality 914:998 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 4 76 theme Drosophila 703:712 arg1 model 721:725 the first Drosophila CDG-Ia model 693:725 the first Drosophila CDG-Ia model 693:725 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 5 77 theme progressive 953:963 arg1 incoordination 965:978 progressive incoordination 953:978 progressive incoordination 953:978 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 1 78 dep production 272:281 arg1 the 268:270 the 268:270 the 268:270 Congenital disorders of glycosylation (CDGs) constitute a rapidly growing family of human diseases resulting from heritable mutations in genes driving the production and modification of glycoproteins. 26940433 4 79 theme model 721:725 arg1 generation 658:667 generation 658:667 generation 658:667 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 4 79 theme model 721:725 arg1 characterization 673:688 characterization 673:688 characterization 673:688 Here, we report the generation and characterization of the first Drosophila CDG-Ia model. 26940433 0 80 theme glycosylation 88:100 arg1 model 110:114 glycosylation disease model 88:114 glycosylation disease model 88:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 9 81 theme Dlp 1735:1737 arg1 co-receptor 1739:1749 Dlp co-receptor 1735:1749 Dlp co-receptor 1735:1749 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 0 82 theme model 110:114 arg1 disorder 76:83 a new Drosophila congenital disorder 48:83 a new Drosophila congenital disorder of glycosylation disease model 48:114 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 5 83 theme shortened 1046:1054 arg1 lifespan 1056:1063 lifespan 1056:1063 lifespan 1056:1063 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 5 84 theme disrupted 791:799 arg1 glycosylation 801:813 severely disrupted glycosylation 782:813 severely disrupted glycosylation 782:813 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 5 85 theme neuronal 871:878 arg1 PMM2 880:883 neuronal PMM2 871:883 neuronal PMM2 871:883 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 86 theme neuromuscular 1167:1179 arg1 NMJ 1191:1193 NMJ 1191:1193 NMJ 1191:1193 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 6 86 theme neuromuscular 1167:1179 arg1 junction 1181:1188 the well-characterized Drosophila neuromuscular junction 1133:1188 the well-characterized Drosophila neuromuscular junction (NMJ) 1133:1194 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 2 87 theme regulated 495:503 arg1 glycosylation 505:517 tightly regulated glycosylation 487:517 tightly regulated glycosylation 487:517 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 8 88 theme synaptic 1678:1685 arg1 activity 1691:1698 synaptic MMP activity 1678:1698 synaptic MMP activity 1678:1698 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 2 89 theme neurological 411:422 arg1 impairments 424:434 severe neurological impairments 404:434 severe neurological impairments 404:434 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 0 90 theme phosphomannomutase 19:36 arg1 type 38:41 phosphomannomutase type 2 19:43 phosphomannomutase type 2 19:43 Synaptic roles for phosphomannomutase type 2 in a new Drosophila congenital disorder of glycosylation disease model. 26940433 9 91 theme co-receptor 1739:1749 arg1 Loss 1701:1704 Loss 1701:1704 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling 1701:1789 Loss of synaptic MMP2, Wg ligand, Dlp co-receptor and downstream trans-synaptic signaling occurs with PMM2 knockdown. 26940433 5 92 theme human 1021:1025 arg1 symptoms 1034:1041 human CDG-Ia symptoms 1021:1041 human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development 1021:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 93 theme well-characterized 1137:1154 arg1 NMJ 1191:1193 NMJ 1191:1193 NMJ 1191:1193 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 6 93 theme well-characterized 1137:1154 arg1 junction 1181:1188 the well-characterized Drosophila neuromuscular junction 1133:1188 the well-characterized Drosophila neuromuscular junction (NMJ) 1133:1194 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 2 94 theme critical 462:469 arg1 requirement 471:481 a particularly critical requirement 447:481 a particularly critical requirement for tightly regulated glycosylation in the nervous system 447:539 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 5 95 theme defective 1091:1099 arg1 development 1108:1118 defective neural development 1091:1118 defective neural development 1091:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 2 96 theme nervous 526:532 arg1 system 534:539 the nervous system 522:539 the nervous system 522:539 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 8 97 theme protein 1545:1551 arg1 Wg 1563:1564 Wg 1563:1564 Wg 1563:1564 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 8 97 theme protein 1545:1551 arg1 Wingless 1553:1560 the Wnt protein Wingless 1537:1560 the Wnt protein Wingless (Wg) 1537:1565 Specifically, trans-synaptic signaling by the Wnt protein Wingless (Wg) depends on the heparan sulfate proteoglycan (HSPG) co-receptor Dally-like protein (Dlp), which is regulated by synaptic MMP activity. 26940433 7 98 theme NMJ 1320:1322 arg1 synaptogenesis 1324:1337 NMJ synaptogenesis 1320:1337 NMJ synaptogenesis 1320:1337 NMJ synaptogenesis is driven by intercellular signals that traverse an extracellular synaptomatrix and are co-regulated by glycosylation and matrix metalloproteinases (MMPs). 26940433 5 99 theme development 1108:1118 arg1 symptoms 1034:1041 human CDG-Ia symptoms 1021:1041 human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development 1021:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 3 100 theme common 551:556 arg1 CDG-Ia 563:568 CDG-Ia 563:568 CDG-Ia (PMM2-CDG) 563:579 The most common CDG, CDG-Ia (PMM2-CDG), arises from phosphomannomutase type 2 (PMM2) mutations. 26940433 3 100 theme common 551:556 arg1 CDG 558:560 The most common CDG 542:560 The most common CDG 542:560 The most common CDG, CDG-Ia (PMM2-CDG), arises from phosphomannomutase type 2 (PMM2) mutations. 26940433 2 101 theme symptomatic 332:342 arg1 hypoglycosylation 344:360 The resulting symptomatic hypoglycosylation 318:360 The resulting symptomatic hypoglycosylation 318:360 The resulting symptomatic hypoglycosylation causes multisystemic defects that include severe neurological impairments, revealing a particularly critical requirement for tightly regulated glycosylation in the nervous system. 26940433 5 102 theme impairments 1075:1085 arg1 symptoms 1034:1041 human CDG-Ia symptoms 1021:1041 human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development 1021:1118 CRISPR-generated pmm2-null Drosophila mutants display severely disrupted glycosylation and early lethality, whereas RNAi-targeted knockdown of neuronal PMM2 results in a strong shift in the abundance of pauci-mannose glycan, progressive incoordination and later lethality, closely paralleling human CDG-Ia symptoms of shortened lifespan, movement impairments and defective neural development. 26940433 6 103 theme synaptic 1203:1210 arg1 loss 1226:1229 synaptic glycosylation loss 1203:1229 synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission 1203:1317 Analyses of the well-characterized Drosophila neuromuscular junction (NMJ) reveal synaptic glycosylation loss accompanied by defects in both structural architecture and functional neurotransmission. 26940433 10 104 theme treatment 2046:2054 arg1 strategies 2056:2065 novel therapeutic treatment strategies 2028:2065 novel therapeutic treatment strategies 2028:2065 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 26940433 10 105 theme disease 1855:1861 arg1 model 1863:1867 this Drosophila CDG disease model 1835:1867 this Drosophila CDG disease model 1835:1867 Taken together, this Drosophila CDG disease model provides a new avenue for the dissection of cellular and molecular mechanisms underlying neurological impairments and is a means by which to discover and test novel therapeutic treatment strategies. 29155734 5 0 theme gel 764:766 arg1 system 776:781 a modified gel imaging system 753:781 a modified gel imaging system (see Table of Materials) 753:806 Gels can be visualized with a modified gel imaging system (see Table of Materials). 29155734 9 1 theme polysaccharide 1311:1324 arg1 substrates 1326:1335 defined polysaccharide substrates 1303:1335 defined polysaccharide substrates 1303:1335 Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates. 29155734 7 2 theme glycosyl 996:1003 arg1 hydrolases 1005:1014 additional glycosyl hydrolases 985:1014 additional glycosyl hydrolases (e.g., mannosidases and galactosidases) 985:1054 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 3 3 theme glycosyl 502:509 arg1 hydrolases 511:520 glycosyl hydrolases 502:520 glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan) 502:593 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 1 4 theme purified 242:249 arg1 material 251:258 purified material 242:258 purified material 242:258 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 5 5 theme imaging 768:774 arg1 system 776:781 a modified gel imaging system 753:781 a modified gel imaging system (see Table of Materials) 753:806 Gels can be visualized with a modified gel imaging system (see Table of Materials). 29155734 0 6 from Characterization 11:26 arg1 Plants 67:72 Plants 67:72 Plants Using PACE 67:83 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 1 7 theme most 160:163 arg1 methods 165:171 most methods 160:171 most methods 160:171 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 1 8 theme material 251:258 arg1 amounts 231:237 large amounts 225:237 large amounts of purified material 225:258 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 1 8 theme material 251:258 arg1 material 251:258 purified material 242:258 purified material 242:258 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 1 8 theme material 251:258 arg1 equipment 191:199 expensive equipment 181:199 expensive equipment 181:199 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 1 8 theme material 251:258 arg1 operators 210:218 skilled operators 202:218 skilled operators 202:218 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 3 9 theme interest 556:563 arg1 polysaccharide 538:551 the polysaccharide 534:551 the polysaccharide of interest 534:563 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 8 10 theme characterized 1184:1196 arg1 hydrolases 1207:1216 characterized glycosyl hydrolases 1184:1216 characterized glycosyl hydrolases 1184:1216 Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. 29155734 3 11 theme wall 469:472 arg1 material 474:481 plant cell wall material 458:481 plant cell wall material 458:481 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 1 12 theme expensive 181:189 arg1 equipment 191:199 expensive equipment 181:199 expensive equipment 181:199 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 7 13 theme branching 938:946 arg1 information 948:958 branching information 938:958 branching information 938:958 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 4 14 theme Large 596:600 arg1 gels 624:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. 29155734 7 15 dep mannosidases 1023:1034 arg1 e.g. 1017:1020 e.g. 1017:1020 e.g. 1017:1020 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 5 16 dep system 776:781 arg1 see 784:786 see 784:786 see Table of Materials 784:805 Gels can be visualized with a modified gel imaging system (see Table of Materials). 29155734 1 17 theme Plant 86:90 arg1 polysaccharides 102:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 2 18 theme isomers 390:396 arg1 resolution 365:374 resolution 365:374 resolution of structural isomers 365:396 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 0 19 theme Structural 0:9 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE 0:83 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 1 20 theme cell 92:95 arg1 polysaccharides 102:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 2 21 theme structural 379:388 arg1 isomers 390:396 structural isomers 379:396 structural isomers 379:396 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 0 22 theme Mannan 31:36 arg1 Polysaccharides 48:62 Mannan Cell Wall Polysaccharides 31:62 Mannan Cell Wall Polysaccharides 31:62 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 5 23 theme Materials 797:805 arg1 Table 788:792 Table 788:792 Table of Materials 788:805 Gels can be visualized with a modified gel imaging system (see Table of Materials). 29155734 1 24 theme wall 97:100 arg1 polysaccharides 102:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides 86:116 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 3 25 theme cell 464:467 arg1 material 474:481 plant cell wall material 458:481 plant cell wall material 458:481 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 3 26 dep hydrolases 511:520 arg1 mannanases 572:581 mannanases 572:581 mannanases for mannan 572:592 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 2 27 theme simple 281:286 arg1 method 288:293 a simple method 279:293 a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers 279:396 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 3 28 dep mannanases 572:581 arg1 e.g. 566:569 e.g. 566:569 e.g. 566:569 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 1 29 theme skilled 202:208 arg1 operators 210:218 skilled operators 202:218 skilled operators 202:218 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 0 30 theme Wall 43:46 arg1 Polysaccharides 48:62 Mannan Cell Wall Polysaccharides 31:62 Mannan Cell Wall Polysaccharides 31:62 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 8 31 theme glucomannan 1111:1121 arg1 structure 1123:1131 glucomannan structure 1111:1131 glucomannan structure 1111:1131 Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. 29155734 4 32 theme released 659:666 arg1 oligosaccharides 668:683 the released oligosaccharides 655:683 the released oligosaccharides 655:683 Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. 29155734 7 33 dep hydrolases 1005:1014 arg1 galactosidases 1040:1053 galactosidases 1040:1053 galactosidases 1040:1053 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 7 33 dep hydrolases 1005:1014 arg1 mannosidases 1023:1034 mannosidases 1023:1034 mannosidases 1023:1034 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 0 34 theme Cell 38:41 arg1 Polysaccharides 48:62 Mannan Cell Wall Polysaccharides 31:62 Mannan Cell Wall Polysaccharides 31:62 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 3 35 theme gel 430:432 arg1 PACE 451:454 PACE 451:454 PACE 451:454 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 3 35 theme gel 430:432 arg1 electrophoresis 434:448 gel electrophoresis 430:448 gel electrophoresis (PACE) 430:455 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 3 36 theme plant 458:462 arg1 material 474:481 plant cell wall material 458:481 plant cell wall material 458:481 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 3 37 theme polysaccharide 403:416 arg1 analysis 418:425 polysaccharide analysis 403:425 polysaccharide analysis by gel electrophoresis (PACE) 403:455 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 0 38 theme Polysaccharides 48:62 arg1 Characterization 11:26 Structural Characterization 0:26 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE 0:83 Structural Characterization of Mannan Cell Wall Polysaccharides in Plants Using PACE. 29155734 9 39 theme novel 1271:1275 arg1 hydrolases 1286:1295 novel glycosyl hydrolases 1271:1295 novel glycosyl hydrolases using defined polysaccharide substrates 1271:1335 Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates. 29155734 9 40 theme glycosyl 1277:1284 arg1 hydrolases 1286:1295 novel glycosyl hydrolases 1271:1295 novel glycosyl hydrolases using defined polysaccharide substrates 1271:1335 Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates. 29155734 2 41 theme structural 331:340 arg1 information 342:352 detailed polysaccharide structural information 307:352 detailed polysaccharide structural information 307:352 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 9 42 used used 1250:1253 arg2 it 1240:1241 it 1240:1241 it 1240:1241 Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates. 29155734 6 43 theme oligosaccharide 823:837 arg1 fingerprint 839:849 The resulting oligosaccharide fingerprint 809:849 The resulting oligosaccharide fingerprint 809:849 The resulting oligosaccharide fingerprint can either be compared qualitatively or, with replication, quantitatively. 29155734 2 44 theme polysaccharide 316:329 arg1 information 342:352 detailed polysaccharide structural information 307:352 detailed polysaccharide structural information 307:352 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 4 45 theme format 602:607 arg1 gels 624:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. 29155734 6 46 theme resulting 813:821 arg1 fingerprint 839:849 The resulting oligosaccharide fingerprint 809:849 The resulting oligosaccharide fingerprint 809:849 The resulting oligosaccharide fingerprint can either be compared qualitatively or, with replication, quantitatively. 29155734 8 47 theme glycosyl 1198:1205 arg1 hydrolases 1207:1216 characterized glycosyl hydrolases 1184:1216 characterized glycosyl hydrolases 1184:1216 Whilst this protocol describes a method for analyzing glucomannan structure, it can be applied to any polysaccharide for which characterized glycosyl hydrolases exist. 29155734 4 48 used used 638:641 arg2 gels 624:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. 29155734 2 49 theme detailed 307:314 arg1 information 342:352 detailed polysaccharide structural information 307:352 detailed polysaccharide structural information 307:352 Here, we describe a simple method for gaining detailed polysaccharide structural information, including resolution of structural isomers. 29155734 1 50 theme large 225:229 arg1 amounts 231:237 large amounts 225:237 large amounts of purified material 225:258 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 1 50 theme large 225:229 arg1 material 251:258 purified material 242:258 purified material 242:258 Plant cell wall polysaccharides are notoriously difficult to analyze, and most methods require expensive equipment, skilled operators, and large amounts of purified material. 29155734 3 51 theme specific 522:529 arg1 hydrolases 511:520 glycosyl hydrolases 502:520 glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan) 502:593 For polysaccharide analysis by gel electrophoresis (PACE), plant cell wall material is hydrolyzed with glycosyl hydrolases specific to the polysaccharide of interest (e.g., mannanases for mannan). 29155734 5 52 theme modified 755:762 arg1 system 776:781 a modified gel imaging system 753:781 a modified gel imaging system (see Table of Materials) 753:806 Gels can be visualized with a modified gel imaging system (see Table of Materials). 29155734 9 53 theme defined 1303:1309 arg1 substrates 1326:1335 defined polysaccharide substrates 1303:1335 defined polysaccharide substrates 1303:1335 Alternatively, it can be used to characterize novel glycosyl hydrolases using defined polysaccharide substrates. 29155734 7 54 theme additional 985:994 arg1 hydrolases 1005:1014 additional glycosyl hydrolases 985:1014 additional glycosyl hydrolases (e.g., mannosidases and galactosidases) 985:1054 Linkage and branching information can be established using additional glycosyl hydrolases (e.g., mannosidases and galactosidases). 29155734 4 55 theme polyacrylamide 609:622 arg1 gels 624:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels 596:627 Large format polyacrylamide gels are then used to separate the released oligosaccharides, which have been fluorescently labeled. 25132658 4 0 theme sucrose 738:744 arg1 supplementation 719:733 the supplementation 715:733 the supplementation of sucrose and tagatose 715:757 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 3 1 theme recombinant 545:555 arg1 protein 557:563 a recombinant protein therapeutic 543:575 a recombinant protein therapeutic 543:575 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 8 2 from tool 1478:1481 arg1 toolbox 1503:1509 the cell culture toolbox 1486:1509 the cell culture toolbox 1486:1509 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 6 3 theme latter 1063:1068 arg1 effect 1070:1075 The latter effect 1059:1075 The latter effect 1059:1075 The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. 25132658 8 4 theme glycosylation 1415:1427 arg1 profiles 1429:1436 protein glycosylation profiles 1407:1436 protein glycosylation profiles 1407:1436 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 8 5 theme sugars 1372:1377 arg1 supplementation 1337:1351 the selective supplementation 1323:1351 the selective supplementation of these described sugars 1323:1377 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 4 6 from profiles 836:843 arg1 proteins 860:867 recombinant proteins 848:867 recombinant proteins 848:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 3 7 theme protein 557:563 arg1 developer 530:538 the developer 526:538 the developer of a recombinant protein therapeutic 526:575 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 1 8 theme recombinant 286:296 arg1 therapeutics 298:309 recombinant therapeutics 286:309 recombinant therapeutics 286:309 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 0 9 theme targeted 90:97 arg1 shifting 99:106 the targeted shifting 86:106 the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics 86:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 7 10 from scales 1252:1257 arg1 reproducible 1226:1237 reproducible 1226:1237 reproducible 1226:1237 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 3 11 theme therapeutic 565:575 arg1 protein 557:563 a recombinant protein therapeutic 543:575 a recombinant protein therapeutic 543:575 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 2 12 theme oligosaccharides 328:343 arg1 addition 316:323 The addition 312:323 The addition of oligosaccharides to recombinant proteins 312:367 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 6 13 theme antibody-dependent 1106:1123 arg1 cytotoxicity 1139:1150 antibody-dependent cell-mediated cytotoxicity 1106:1150 antibody-dependent cell-mediated cytotoxicity 1106:1150 The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. 25132658 1 14 theme therapeutics 298:309 arg1 function 274:281 function 274:281 function 274:281 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 1 14 theme therapeutics 298:309 arg1 structure 260:268 structure 260:268 structure 260:268 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 7 15 theme cell 1303:1306 arg1 lines 1308:1312 different Chinese hamster ovary cell lines 1271:1312 different Chinese hamster ovary cell lines 1271:1312 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 5 16 theme concomitant 1020:1030 arg1 reduction 1032:1040 a concomitant reduction 1018:1040 a concomitant reduction in fucosylation 1018:1056 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 4 17 theme tagatose 750:757 arg1 supplementation 719:733 the supplementation 715:733 the supplementation of sucrose and tagatose 715:757 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 5 18 theme species 998:1004 arg1 percentage 962:971 the percentage 958:971 the percentage of high mannose N-glycan species 958:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 7 19 theme Chinese 1281:1287 arg1 lines 1308:1312 different Chinese hamster ovary cell lines 1271:1312 different Chinese hamster ovary cell lines 1271:1312 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 5 20 theme high 976:979 arg1 species 998:1004 high mannose N-glycan species 976:1004 high mannose N-glycan species 976:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 7 21 theme aforementioned 1186:1199 arg1 results 1201:1207 These aforementioned results 1180:1207 These aforementioned results 1180:1207 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 6 22 theme cell-mediated 1125:1137 arg1 cytotoxicity 1139:1150 antibody-dependent cell-mediated cytotoxicity 1106:1150 antibody-dependent cell-mediated cytotoxicity 1106:1150 The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. 25132658 4 23 theme profiles 836:843 arg1 modulation 800:809 the targeted modulation 787:809 the targeted modulation of protein glycosylation profiles on recombinant proteins 787:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 1 24 theme Protein 179:185 arg1 modification 236:247 an important post-translational modification 204:247 an important post-translational modification toward the structure and function of recombinant therapeutics 204:309 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 1 24 theme Protein 179:185 arg1 glycosylation 187:199 Protein glycosylation 179:199 Protein glycosylation 179:199 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 7 25 theme hamster 1289:1295 arg1 lines 1308:1312 different Chinese hamster ovary cell lines 1271:1312 different Chinese hamster ovary cell lines 1271:1312 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 8 26 theme profiles 1429:1436 arg1 modulation 1393:1402 the targeted modulation 1380:1402 the targeted modulation of protein glycosylation profiles 1380:1436 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 7 27 theme different 1271:1279 arg1 lines 1308:1312 different Chinese hamster ovary cell lines 1271:1312 different Chinese hamster ovary cell lines 1271:1312 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 4 28 from modulation 800:809 arg1 proteins 860:867 recombinant proteins 848:867 recombinant proteins 848:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 4 29 theme glycosylation 822:834 arg1 profiles 836:843 protein glycosylation profiles 814:843 protein glycosylation profiles on recombinant proteins 814:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 2 30 theme many 446:449 arg1 proteins 451:458 many proteins 446:458 many proteins 446:458 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 0 31 theme culture 5:11 arg1 media 13:17 Cell culture media 0:17 Cell culture media 0:17 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 32 theme Cell 0:3 arg1 media 13:17 Cell culture media 0:17 Cell culture media 0:17 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 4 33 theme protein 814:820 arg1 profiles 836:843 protein glycosylation profiles 814:843 protein glycosylation profiles on recombinant proteins 814:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 6 34 theme recombinant 1158:1168 arg1 antibody 1170:1177 a recombinant antibody 1156:1177 a recombinant antibody 1156:1177 The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. 25132658 0 35 theme glycosylation 119:131 arg1 profiles 133:140 protein glycosylation profiles 111:140 protein glycosylation profiles of recombinant protein therapeutics 111:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 8 36 theme culture 1495:1501 arg1 toolbox 1503:1509 the cell culture toolbox 1486:1509 the cell culture toolbox 1486:1509 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 2 37 theme recombinant 348:358 arg1 proteins 360:367 recombinant proteins 348:367 recombinant proteins 348:367 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 8 38 theme cell 1490:1493 arg1 toolbox 1503:1509 the cell culture toolbox 1486:1509 the cell culture toolbox 1486:1509 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 1 39 theme important 207:215 arg1 modification 236:247 an important post-translational modification 204:247 an important post-translational modification toward the structure and function of recombinant therapeutics 204:309 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 1 39 theme important 207:215 arg1 glycosylation 187:199 Protein glycosylation 179:199 Protein glycosylation 179:199 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 5 40 from increase 946:953 arg1 percentage 962:971 the percentage 958:971 the percentage of high mannose N-glycan species 958:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 5 40 from increase 946:953 arg1 fucosylation 1045:1056 fucosylation 1045:1056 fucosylation 1045:1056 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 0 41 theme protein 111:117 arg1 profiles 133:140 protein glycosylation profiles 111:140 protein glycosylation profiles of recombinant protein therapeutics 111:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 4 42 theme targeted 791:798 arg1 modulation 800:809 the targeted modulation 787:809 the targeted modulation of protein glycosylation profiles on recombinant proteins 787:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 1 43 theme post-translational 217:234 arg1 modification 236:247 an important post-translational modification 204:247 an important post-translational modification toward the structure and function of recombinant therapeutics 204:309 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 1 43 theme post-translational 217:234 arg1 glycosylation 187:199 Protein glycosylation 179:199 Protein glycosylation 179:199 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 7 44 theme different 1242:1250 arg1 scales 1252:1257 different scales 1242:1257 different scales 1242:1257 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 5 45 theme mannose 981:987 arg1 species 998:1004 high mannose N-glycan species 976:1004 high mannose N-glycan species 976:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 5 46 theme oligosaccharide 903:917 arg1 maturation 919:928 oligosaccharide maturation 903:928 oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation 903:1056 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 0 47 dep sugars 54:59 arg1 tagatose 73:80 tagatose 73:80 tagatose 73:80 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 47 dep sugars 54:59 arg1 sucrose 61:67 sucrose 61:67 sucrose 61:67 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 47 dep sugars 54:59 arg1 sugars 54:59 uncommonly used sugars 38:59 uncommonly used sugars sucrose and tagatose 38:80 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 4 48 theme systematic 691:700 arg1 study 702:706 a systematic study 689:706 a systematic study toward the supplementation of sucrose and tagatose 689:757 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 2 49 theme proteins 451:458 arg1 attributes 432:441 the overall physiochemical attributes 405:441 the overall physiochemical attributes of many proteins 405:458 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 0 50 theme profiles 133:140 arg1 shifting 99:106 the targeted shifting 86:106 the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics 86:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 8 51 theme described 1362:1370 arg1 sugars 1372:1377 these described sugars 1356:1377 these described sugars 1356:1377 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 0 52 theme sugars 54:59 arg1 supplementation 19:33 supplementation 19:33 supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics 19:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 7 53 theme ovary 1297:1301 arg1 lines 1308:1312 different Chinese hamster ovary cell lines 1271:1312 different Chinese hamster ovary cell lines 1271:1312 These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. 25132658 0 54 theme protein 157:163 arg1 therapeutics 165:176 recombinant protein therapeutics 145:176 recombinant protein therapeutics 145:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 1 55 dep structure 260:268 arg1 the 256:258 the 256:258 the 256:258 Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. 25132658 3 56 theme critical 634:641 arg1 attribute 651:659 a critical quality attribute 632:659 a critical quality attribute 632:659 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 4 57 theme cell 764:767 arg1 media 777:781 cell culture media 764:781 cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins 764:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 0 58 theme used 49:52 arg1 tagatose 73:80 tagatose 73:80 tagatose 73:80 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 58 theme used 49:52 arg1 sucrose 61:67 sucrose 61:67 sucrose 61:67 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 58 theme used 49:52 arg1 sugars 54:59 uncommonly used sugars 38:59 uncommonly used sugars sucrose and tagatose 38:80 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 0 59 theme recombinant 145:155 arg1 therapeutics 165:176 recombinant protein therapeutics 145:176 recombinant protein therapeutics 145:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 3 60 theme quality 643:649 arg1 attribute 651:659 a critical quality attribute 632:659 a critical quality attribute 632:659 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 0 61 gly glycosylation 119:131 arg1 therapeutics 165:176 recombinant protein therapeutics 145:176 recombinant protein therapeutics 145:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 4 62 theme recombinant 848:858 arg1 proteins 860:867 recombinant proteins 848:867 recombinant proteins 848:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 0 63 dep supplementation 19:33 arg1 media 13:17 Cell culture media 0:17 Cell culture media 0:17 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 5 64 theme N-glycan 989:996 arg1 species 998:1004 high mannose N-glycan species 976:1004 high mannose N-glycan species 976:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 0 65 theme therapeutics 165:176 arg1 profiles 133:140 protein glycosylation profiles 111:140 protein glycosylation profiles of recombinant protein therapeutics 111:176 Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics. 25132658 3 66 theme protein 586:592 arg1 glycosylation 594:606 protein glycosylation 586:606 protein glycosylation 586:606 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 4 67 theme culture 769:775 arg1 media 777:781 cell culture media 764:781 cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins 764:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 8 68 theme protein 1407:1413 arg1 profiles 1429:1436 protein glycosylation profiles 1407:1436 protein glycosylation profiles 1407:1436 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 8 69 theme targeted 1384:1391 arg1 modulation 1393:1402 the targeted modulation 1380:1402 the targeted modulation of protein glycosylation profiles 1380:1436 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 4 70 from proteins 860:867 arg1 modulation 800:809 the targeted modulation 787:809 the targeted modulation of protein glycosylation profiles on recombinant proteins 787:867 In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. 25132658 2 71 theme physiochemical 417:430 arg1 attributes 432:441 the overall physiochemical attributes 405:441 the overall physiochemical attributes of many proteins 405:458 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 3 72 theme protein 488:494 arg1 glycosylation 496:508 protein glycosylation 488:508 protein glycosylation 488:508 It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. 25132658 8 73 theme selective 1327:1335 arg1 supplementation 1337:1351 the selective supplementation 1323:1351 the selective supplementation of these described sugars 1323:1377 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 8 74 theme product 1524:1530 arg1 comparability 1532:1544 product comparability 1524:1544 product comparability 1524:1544 Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. 25132658 2 75 theme overall 409:415 arg1 attributes 432:441 the overall physiochemical attributes 405:441 the overall physiochemical attributes of many proteins 405:458 The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. 25132658 5 76 from reduction 1032:1040 arg1 percentage 962:971 the percentage 958:971 the percentage of high mannose N-glycan species 958:1004 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 25132658 5 76 from reduction 1032:1040 arg1 fucosylation 1045:1056 fucosylation 1045:1056 fucosylation 1045:1056 Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. 28334934 7 0 theme teichoic 1345:1352 arg1 acid 1354:1357 teichoic acid 1345:1357 teichoic acid biosynthesis 1345:1370 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 10 1 with Cross-complementation 1760:1780 arg1 transferase 1833:1843 the nonnative nonulosonic acid transferase 1802:1843 the nonnative nonulosonic acid transferase 1802:1843 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 4 2 theme abundant 815:822 arg1 glycoproteins 832:844 the bacterium's abundant S-layer glycoproteins 799:844 the bacterium's abundant S-layer glycoproteins 799:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 5 3 dep genes 893:897 arg1 TFUB4_00887 934:944 TFUB4_00887 934:944 TFUB4_00887 934:944 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 3 dep genes 893:897 arg1 UB4 954:956 strain UB4 947:956 strain UB4 947:956 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 3 dep genes 893:897 arg1 Tanf_01245 899:908 Tanf_01245 899:908 Tanf_01245 899:908 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 3 dep genes 893:897 arg1 ATCC 918:921 strain ATCC 43037 911:927 strain ATCC 43037 911:927 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 3 dep genes 893:897 arg1 genes 893:897 the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4) 879:957 the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4) 879:957 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 4 4 theme acid 738:741 arg1 residue 743:749 a proximal N-acetylmannosaminuronic acid residue 702:749 a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins 702:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 11 5 theme legionaminic 1974:1985 arg1 transferase 2003:2013 a legionaminic acid derivative transferase 1972:2013 a legionaminic acid derivative transferase 1972:2013 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 2 6 theme acid 333:336 arg1 transferases 338:349 cognate nonulosonic acid transferases 313:349 cognate nonulosonic acid transferases 313:349 However, the knowledge of cognate nonulosonic acid transferases is scarce. 28334934 9 7 theme gene 1592:1595 arg1 mutants 1607:1613 Single gene knock-out mutants 1585:1613 Single gene knock-out mutants targeted at either transferase 1585:1644 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 0 8 theme O-glycosylation 122:136 arg1 system 138:143 the general protein O-glycosylation system 102:143 the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia 102:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 2 9 theme cognate 313:319 arg1 transferases 338:349 cognate nonulosonic acid transferases 313:349 cognate nonulosonic acid transferases 313:349 However, the knowledge of cognate nonulosonic acid transferases is scarce. 28334934 1 10 theme acids 225:229 arg1 occurrence 199:208 The occurrence 195:208 The occurrence of nonulosonic acids in bacteria 195:241 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 1 10 theme acids 225:229 arg1 wide-spread 246:256 wide-spread 246:256 wide-spread 246:256 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 4 11 theme proximal 704:711 arg1 residue 743:749 a proximal N-acetylmannosaminuronic acid residue 702:749 a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins 702:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 11 12 theme derivative 1992:2001 arg1 transferase 2003:2013 a legionaminic acid derivative transferase 1972:2013 a legionaminic acid derivative transferase 1972:2013 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 0 13 theme periodontal 152:162 arg1 forsythia 184:192 the periodontal pathogen Tannerella forsythia 148:192 the periodontal pathogen Tannerella forsythia 148:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 8 14 theme sugars 1425:1430 arg1 pool 1396:1399 a cellular pool 1385:1399 a cellular pool of nucleotide-activated sugars 1385:1430 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 0 15 theme Tannerella 173:182 arg1 forsythia 184:192 the periodontal pathogen Tannerella forsythia 148:192 the periodontal pathogen Tannerella forsythia 148:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 1 16 from occurrence 199:208 arg1 bacteria 234:241 bacteria 234:241 bacteria 234:241 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 5 17 theme acid 1011:1014 arg1 derivative 1016:1025 a legionaminic acid derivative 996:1025 a legionaminic acid derivative 996:1025 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 4 18 theme O-glycan 762:769 arg1 structure 771:779 the O-glycan structure 758:779 the O-glycan structure 758:779 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 5 19 theme Bioinformatic 847:859 arg1 analyses 861:868 Bioinformatic analyses 847:868 Bioinformatic analyses 847:868 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 9 20 theme S-layer 1664:1670 arg1 composition 1681:1691 S-layer O-glycan composition 1664:1691 S-layer O-glycan composition 1664:1691 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 7 21 theme N-acetylmannosamine 1314:1332 arg1 residue 1334:1340 an N-acetylmannosamine residue 1311:1340 an N-acetylmannosamine residue in teichoic acid biosynthesis 1311:1370 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 5 22 theme pseudaminic 980:990 arg1 transferase 1027:1037 a putative pseudaminic and a legionaminic acid derivative transferase 969:1037 a putative pseudaminic and a legionaminic acid derivative transferase 969:1037 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 3 23 theme Tannerella 388:397 arg1 forsythia 399:407 the periodontopathogen Tannerella forsythia 365:407 the periodontopathogen Tannerella forsythia 365:407 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 8 24 theme CMP-activated 1462:1474 arg1 substrates 1539:1548 substrates 1539:1548 substrates for the corresponding transferase 1539:1582 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 8 24 theme CMP-activated 1462:1474 arg1 derivatives 1493:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 4 25 theme S-layer 824:830 arg1 glycoproteins 832:844 the bacterium's abundant S-layer glycoproteins 799:844 the bacterium's abundant S-layer glycoproteins 799:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 6 26 theme identical 1078:1086 arg1 C-termini 1088:1096 identical C-termini 1078:1096 identical C-termini 1078:1096 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 8 27 theme acid 1488:1491 arg1 substrates 1539:1548 substrates 1539:1548 substrates for the corresponding transferase 1539:1582 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 8 27 theme acid 1488:1491 arg1 derivatives 1493:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 10 28 theme nonulosonic 1816:1826 arg1 transferase 1833:1843 the nonnative nonulosonic acid transferase 1802:1843 the nonnative nonulosonic acid transferase 1802:1843 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 7 29 theme glycerol 1290:1297 arg1 transfer 1299:1306 glycerol transfer 1290:1306 glycerol transfer 1290:1306 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 3 30 theme proposed 418:425 arg1 factors 437:443 several proposed virulence factors 410:443 several proposed virulence factors 410:443 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 31 dep strain-specifically 451:469 arg1 either 471:476 either 471:476 either 471:476 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 6 32 dep sialyltransferases 1169:1186 arg1 sialyltransferases 1169:1186 bacterial sialyltransferases 1159:1186 bacterial sialyltransferases (D/E-D/E-G and HP) 1159:1205 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 32 dep sialyltransferases 1169:1186 arg1 HP 1203:1204 HP 1203:1204 HP 1203:1204 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 32 dep sialyltransferases 1169:1186 arg1 D/E-D/E-G 1189:1197 D/E-D/E-G 1189:1197 D/E-D/E-G 1189:1197 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 11 33 theme novel 2069:2073 arg1 sialoglycoconjugates 2075:2094 novel sialoglycoconjugates 2069:2094 novel sialoglycoconjugates 2069:2094 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 5 34 theme strain 911:916 arg1 Tanf_01245 899:908 Tanf_01245 899:908 Tanf_01245 899:908 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 34 theme strain 911:916 arg1 ATCC 918:921 strain ATCC 43037 911:927 strain ATCC 43037 911:927 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 6 35 theme bacterial 1159:1167 arg1 sialyltransferases 1169:1186 bacterial sialyltransferases 1159:1186 bacterial sialyltransferases (D/E-D/E-G and HP) 1159:1205 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 35 theme bacterial 1159:1167 arg1 HP 1203:1204 HP 1203:1204 HP 1203:1204 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 35 theme bacterial 1159:1167 arg1 D/E-D/E-G 1189:1197 D/E-D/E-G 1189:1197 D/E-D/E-G 1189:1197 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 0 36 theme acid 14:17 arg1 transferase 53:63 A pseudaminic acid or a legionaminic acid derivative transferase 0:63 A pseudaminic acid or a legionaminic acid derivative transferase 0:63 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 3 37 theme legionaminic 497:508 arg1 derivative 515:524 a legionaminic acid derivative 495:524 a legionaminic acid derivative 495:524 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 5 38 theme candidate 883:891 arg1 genes 893:897 the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4) 879:957 the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4) 879:957 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 38 theme candidate 883:891 arg1 TFUB4_00887 934:944 TFUB4_00887 934:944 TFUB4_00887 934:944 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 38 theme candidate 883:891 arg1 Tanf_01245 899:908 Tanf_01245 899:908 Tanf_01245 899:908 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 9 39 theme acid 1743:1746 arg1 derivative 1748:1757 the nonulosonic acid derivative 1727:1757 the nonulosonic acid derivative 1727:1757 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 0 40 theme acid 37:40 arg1 derivative 42:51 a legionaminic acid derivative 22:51 a legionaminic acid derivative 22:51 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 3 41 theme terminal 529:536 arg1 sugar 538:542 terminal sugar 529:542 terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide 529:612 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 42 theme acid 510:513 arg1 derivative 515:524 a legionaminic acid derivative 495:524 a legionaminic acid derivative 495:524 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 8 43 theme corresponding 1558:1570 arg1 transferase 1572:1582 the corresponding transferase 1554:1582 the corresponding transferase 1554:1582 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 3 44 theme identical 573:581 arg1 oligosaccharide 598:612 an otherwise structurally identical, protein-bound oligosaccharide 547:612 an otherwise structurally identical, protein-bound oligosaccharide 547:612 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 0 45 theme derivative 42:51 arg1 transferase 53:63 A pseudaminic acid or a legionaminic acid derivative transferase 0:63 A pseudaminic acid or a legionaminic acid derivative transferase 0:63 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 4 46 theme nonulosonic 671:681 arg1 derivative 688:697 either nonulosonic acid derivative 664:697 either nonulosonic acid derivative 664:697 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 3 47 contain carry 445:449 arg3 forsythia 399:407 the periodontopathogen Tannerella forsythia 365:407 the periodontopathogen Tannerella forsythia 365:407 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 47 contain carry 445:449 arg2 derivative 515:524 a legionaminic acid derivative 495:524 a legionaminic acid derivative 495:524 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 47 contain carry 445:449 arg2 strain-specifically 451:469 strain-specifically 451:469 strain-specifically 451:469 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 47 contain carry 445:449 arg2 pseudaminic 480:490 a pseudaminic 478:490 a pseudaminic 478:490 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 3 47 contain carry 445:449 arg1 factors 437:443 several proposed virulence factors 410:443 several proposed virulence factors 410:443 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 7 48 theme acid 1354:1357 arg1 biosynthesis 1359:1370 teichoic acid biosynthesis 1345:1370 teichoic acid biosynthesis 1345:1370 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 9 49 theme O-glycan 1672:1679 arg1 composition 1681:1691 S-layer O-glycan composition 1664:1691 S-layer O-glycan composition 1664:1691 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 5 50 theme strain 947:952 arg1 TFUB4_00887 934:944 TFUB4_00887 934:944 TFUB4_00887 934:944 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 5 50 theme strain 947:952 arg1 UB4 954:956 strain UB4 947:956 strain UB4 947:956 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 7 51 theme B 1236:1236 arg1 glycosyltransferases 1238:1257 type B glycosyltransferases 1231:1257 type B glycosyltransferases 1231:1257 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 4 52 theme N-acetylmannosaminuronic 713:736 arg1 residue 743:749 a proximal N-acetylmannosaminuronic acid residue 702:749 a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins 702:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 3 53 from sugar 538:542 arg1 oligosaccharide 598:612 an otherwise structurally identical, protein-bound oligosaccharide 547:612 an otherwise structurally identical, protein-bound oligosaccharide 547:612 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 2 54 theme transferases 338:349 arg1 knowledge 300:308 the knowledge 296:308 the knowledge of cognate nonulosonic acid transferases 296:349 However, the knowledge of cognate nonulosonic acid transferases is scarce. 28334934 2 54 theme transferases 338:349 arg1 scarce 354:359 scarce 354:359 scarce 354:359 However, the knowledge of cognate nonulosonic acid transferases is scarce. 28334934 9 55 theme Single 1585:1590 arg1 mutants 1607:1613 Single gene knock-out mutants 1585:1613 Single gene knock-out mutants targeted at either transferase 1585:1644 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 11 56 theme acid 1987:1990 arg1 transferase 2003:2013 a legionaminic acid derivative transferase 1972:2013 a legionaminic acid derivative transferase 1972:2013 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 2 57 theme nonulosonic 321:331 arg1 transferases 338:349 cognate nonulosonic acid transferases 313:349 cognate nonulosonic acid transferases 313:349 However, the knowledge of cognate nonulosonic acid transferases is scarce. 28334934 9 58 theme knock-out 1597:1605 arg1 mutants 1607:1613 Single gene knock-out mutants 1585:1613 Single gene knock-out mutants targeted at either transferase 1585:1644 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 4 59 theme derivative 688:697 arg1 transfer 652:659 the transfer 648:659 the transfer of either nonulosonic acid derivative 648:697 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 10 60 theme mutants 1789:1795 arg1 Cross-complementation 1760:1780 Cross-complementation 1760:1780 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase 1760:1843 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 11 61 theme valuable 2035:2042 arg1 tools 2044:2048 valuable tools 2035:2048 valuable tools for engineering of novel sialoglycoconjugates 2035:2094 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 11 61 theme valuable 2035:2042 arg1 these 2016:2020 these 2016:2020 these 2016:2020 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 0 62 theme pathogen 164:171 arg1 forsythia 184:192 the periodontal pathogen Tannerella forsythia 148:192 the periodontal pathogen Tannerella forsythia 148:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 1 63 attach linked 262:267 arg1 pathogenicity 272:284 pathogenicity 272:284 pathogenicity 272:284 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 1 63 attach linked 262:267 arg2 wide-spread 246:256 wide-spread 246:256 wide-spread 246:256 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 1 63 attach linked 262:267 arg2 occurrence 199:208 The occurrence 195:208 The occurrence of nonulosonic acids in bacteria 195:241 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 8 64 theme pool 1396:1399 arg1 Analysis 1373:1380 Analysis 1373:1380 Analysis of a cellular pool of nucleotide-activated sugars 1373:1430 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 0 65 theme forsythia 184:192 arg1 system 138:143 the general protein O-glycosylation system 102:143 the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia 102:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 5 66 theme legionaminic 998:1009 arg1 derivative 1016:1025 a legionaminic acid derivative 996:1025 a legionaminic acid derivative 996:1025 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 6 67 theme glycosyltransferases 1128:1147 arg1 typical 1117:1123 typical 1117:1123 typical 1117:1123 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 7 68 from residue 1334:1340 arg1 biosynthesis 1359:1370 teichoic acid biosynthesis 1345:1370 teichoic acid biosynthesis 1345:1370 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 8 69 theme nucleotide-activated 1404:1423 arg1 sugars 1425:1430 nucleotide-activated sugars 1404:1430 nucleotide-activated sugars 1404:1430 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 11 70 theme plausible 1929:1937 arg1 candidates 1939:1948 plausible candidates 1929:1948 plausible candidates for a pseudaminic and a legionaminic acid derivative transferase 1929:2013 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 1 71 theme nonulosonic 213:223 arg1 acids 225:229 nonulosonic acids 213:229 nonulosonic acids 213:229 The occurrence of nonulosonic acids in bacteria is wide-spread and linked to pathogenicity. 28334934 6 72 theme typical 1117:1123 arg1 motifs 1110:1115 motifs 1110:1115 motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP) 1110:1205 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 5 73 theme putative 971:978 arg1 pseudaminic 980:990 putative pseudaminic 971:990 putative pseudaminic 971:990 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 0 74 theme protein 114:120 arg1 system 138:143 the general protein O-glycosylation system 102:143 the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia 102:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 3 75 theme periodontopathogen 369:386 arg1 forsythia 399:407 the periodontopathogen Tannerella forsythia 365:407 the periodontopathogen Tannerella forsythia 365:407 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 8 76 theme nonulosonic 1476:1486 arg1 substrates 1539:1548 substrates 1539:1548 substrates for the corresponding transferase 1539:1582 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 8 76 theme nonulosonic 1476:1486 arg1 derivatives 1493:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 the CMP-activated nonulosonic acid derivatives 1458:1503 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 10 77 theme nonnative 1806:1814 arg1 transferase 1833:1843 the nonnative nonulosonic acid transferase 1802:1843 the nonnative nonulosonic acid transferase 1802:1843 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 0 78 theme pseudaminic 2:12 arg1 acid 14:17 A pseudaminic acid 0:17 A pseudaminic acid 0:17 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 5 79 theme derivative 1016:1025 arg1 transferase 1027:1037 a putative pseudaminic and a legionaminic acid derivative transferase 969:1037 a putative pseudaminic and a legionaminic acid derivative transferase 969:1037 Bioinformatic analyses provided the candidate genes Tanf_01245 (strain ATCC 43037) and TFUB4_00887 (strain UB4), encoding a putative pseudaminic and a legionaminic acid derivative transferase, respectively. 28334934 6 80 theme sialyltransferases 1169:1186 arg1 typical 1117:1123 typical 1117:1123 typical 1117:1123 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 3 81 theme virulence 427:435 arg1 factors 437:443 several proposed virulence factors 410:443 several proposed virulence factors 410:443 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 8 82 theme derivatives 1493:1503 arg1 presence 1446:1453 the presence 1442:1453 the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase 1442:1582 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 11 83 theme sialoglycoconjugates 2075:2094 arg1 engineering 2054:2064 engineering 2054:2064 engineering of novel sialoglycoconjugates 2054:2094 This study identified plausible candidates for a pseudaminic and a legionaminic acid derivative transferase; these may serve as valuable tools for engineering of novel sialoglycoconjugates. 28334934 3 84 dep identical 573:581 arg1 protein-bound 584:596 protein-bound 584:596 protein-bound 584:596 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 10 85 theme acid 1828:1831 arg1 transferase 1833:1843 the nonnative nonulosonic acid transferase 1802:1843 the nonnative nonulosonic acid transferase 1802:1843 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 3 86 theme several 410:416 arg1 factors 437:443 several proposed virulence factors 410:443 several proposed virulence factors 410:443 In the periodontopathogen Tannerella forsythia, several proposed virulence factors carry strain-specifically either a pseudaminic or a legionaminic acid derivative as terminal sugar on an otherwise structurally identical, protein-bound oligosaccharide. 28334934 6 87 contain contain 1102:1108 arg1 transferases 1060:1071 These transferases 1054:1071 These transferases 1054:1071 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 87 contain contain 1102:1108 arg2 motifs 1110:1115 motifs 1110:1115 motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP) 1110:1205 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 0 88 theme legionaminic 24:35 arg1 derivative 42:51 a legionaminic acid derivative 22:51 a legionaminic acid derivative 22:51 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 8 89 theme cellular 1387:1394 arg1 pool 1396:1399 a cellular pool 1385:1399 a cellular pool of nucleotide-activated sugars 1385:1430 Analysis of a cellular pool of nucleotide-activated sugars confirmed the presence of the CMP-activated nonulosonic acid derivatives, which are most likely serving as substrates for the corresponding transferase. 28334934 9 90 theme nonulosonic 1731:1741 arg1 derivative 1748:1757 the nonulosonic acid derivative 1727:1757 the nonulosonic acid derivative 1727:1757 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 0 91 theme general 106:112 arg1 system 138:143 the general protein O-glycosylation system 102:143 the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia 102:192 A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia. 28334934 7 92 theme type 1231:1234 arg1 glycosyltransferases 1238:1257 type B glycosyltransferases 1231:1257 type B glycosyltransferases 1231:1257 They share homology to type B glycosyltransferases and TagB, an enzyme catalyzing glycerol transfer to an N-acetylmannosamine residue in teichoic acid biosynthesis. 28334934 9 93 theme derivative 1748:1757 arg1 loss 1719:1722 the loss 1715:1722 the loss of the nonulosonic acid derivative 1715:1757 Single gene knock-out mutants targeted at either transferase were analyzed for S-layer O-glycan composition by ESI-MS, confirming the loss of the nonulosonic acid derivative. 28334934 6 94 contain have 1073:1076 arg1 transferases 1060:1071 These transferases 1054:1071 These transferases 1054:1071 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 6 94 contain have 1073:1076 arg2 C-termini 1088:1096 identical C-termini 1078:1096 identical C-termini 1078:1096 These transferases have identical C-termini and contain motifs typical of glycosyltransferases (DXD) and bacterial sialyltransferases (D/E-D/E-G and HP). 28334934 10 95 theme high 1875:1878 arg1 stringency 1880:1889 high stringency 1875:1889 high stringency of the enzymes 1875:1904 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 28334934 4 96 gly glycoproteins 832:844 arg1 glycoproteins 832:844 the bacterium's abundant S-layer glycoproteins 799:844 the bacterium's abundant S-layer glycoproteins 799:844 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 4 97 theme acid 683:686 arg1 derivative 688:697 either nonulosonic acid derivative 664:697 either nonulosonic acid derivative 664:697 This study aims to shed light on the transfer of either nonulosonic acid derivative on a proximal N-acetylmannosaminuronic acid residue within the O-glycan structure, exemplified with the bacterium's abundant S-layer glycoproteins. 28334934 10 98 theme enzymes 1898:1904 arg1 stringency 1880:1889 high stringency 1875:1889 high stringency of the enzymes 1875:1904 Cross-complementation of the mutants with the nonnative nonulosonic acid transferase was not successful indicating high stringency of the enzymes. 25296255 5 0 theme gp41-tryptophan 1003:1017 arg1 clasp 1019:1023 a gp41-tryptophan clasp 1001:1023 a gp41-tryptophan clasp 1001:1023 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 7 1 theme closed 1193:1198 arg1 spike 1200:1204 the pre-fusion closed spike 1178:1204 the pre-fusion closed spike 1178:1204 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 4 2 theme recognition 794:804 arg1 parameters 754:763 parameters 754:763 parameters of immune evasion and immune recognition 754:804 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 2 3 from antigen 369:375 arg1 surface 397:403 the HIV-1 virion surface 380:403 the HIV-1 virion surface 380:403 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 5 4 theme amino- 833:838 arg1 strands 861:867 amino- and carboxy-terminal strands 833:867 amino- and carboxy-terminal strands of gp120 833:876 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 1 5 theme receptor-bound 297:310 arg1 intermediates 312:324 receptor-bound intermediates 297:324 receptor-bound intermediates 297:324 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 4 6 theme immune 787:792 arg1 recognition 794:804 immune recognition 787:804 immune recognition 787:804 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 1 7 theme spike 130:134 arg1 type 100:103 The human immunodeficiency virus type 1 67:105 The human immunodeficiency virus type 1 (HIV-1) 67:113 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 7 theme spike 130:134 arg1 HIV-1 108:112 HIV-1 108:112 HIV-1 108:112 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 7 theme spike 130:134 arg1 machine 205:211 a conformational machine 188:211 a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state 188:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 6 8 dep opening 1074:1080 arg1 expelling 1096:1104 expelling 1096:1104 expelling 1096:1104 Spike rearrangements required for entry involve opening the clasp and expelling the termini. 25296255 6 8 dep opening 1074:1080 arg1 clasp 1086:1090 clasp 1086:1090 clasp 1086:1090 Spike rearrangements required for entry involve opening the clasp and expelling the termini. 25296255 7 9 dep prevalence 1242:1251 arg1 the 1238:1240 the 1238:1240 the 1238:1240 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 4 10 theme pre-fusion 654:663 arg1 conformation 665:676 the pre-fusion conformation 650:676 the pre-fusion conformation of gp41 650:684 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 4 11 theme gp41 681:684 arg1 conformation 665:676 the pre-fusion conformation 650:676 the pre-fusion conformation of gp41 650:684 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 1 12 theme HIV-1 230:234 arg1 entry 236:240 HIV-1 entry 230:240 HIV-1 entry 230:240 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 4 13 theme immune 768:773 arg1 evasion 775:781 immune evasion 768:781 immune evasion 768:781 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 5 14 theme membrane-proximal 908:924 arg1 collar 926:931 a membrane-proximal collar 906:931 a membrane-proximal collar 906:931 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 2 15 theme antibodies 445:454 arg1 Env 406:408 Env 406:408 Env 406:408 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 2 15 theme antibodies 445:454 arg1 target 422:427 the target 418:427 the target of neutralizing antibodies 418:454 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 2 15 theme antibodies 445:454 arg1 focus 462:466 a focus 460:466 a focus of vaccine efforts 460:485 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 1 16 theme human 71:75 arg1 type 100:103 The human immunodeficiency virus type 1 67:105 The human immunodeficiency virus type 1 (HIV-1) 67:113 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 16 theme human 71:75 arg1 HIV-1 108:112 HIV-1 108:112 HIV-1 108:112 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 16 theme human 71:75 arg1 machine 205:211 a conformational machine 188:211 a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state 188:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 6 17 theme Spike 1026:1030 arg1 rearrangements 1032:1045 Spike rearrangements 1026:1045 Spike rearrangements required for entry 1026:1064 Spike rearrangements required for entry involve opening the clasp and expelling the termini. 25296255 1 18 theme post-fusion 332:342 arg1 state 344:348 a post-fusion state 330:348 a post-fusion state 330:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 19 theme immunodeficiency 77:92 arg1 type 100:103 The human immunodeficiency virus type 1 67:105 The human immunodeficiency virus type 1 (HIV-1) 67:113 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 19 theme immunodeficiency 77:92 arg1 HIV-1 108:112 HIV-1 108:112 HIV-1 108:112 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 19 theme immunodeficiency 77:92 arg1 machine 205:211 a conformational machine 188:211 a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state 188:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 7 20 theme responses 1298:1306 arg1 location 1257:1264 location 1257:1264 location 1257:1264 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 20 theme responses 1298:1306 arg1 prevalence 1242:1251 prevalence 1242:1251 prevalence 1242:1251 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 2 21 theme virion 390:395 arg1 surface 397:403 the HIV-1 virion surface 380:403 the HIV-1 virion surface 380:403 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 4 22 theme evasion 775:781 arg1 parameters 754:763 parameters 754:763 parameters of immune evasion and immune recognition 754:804 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 1 23 theme virus 94:98 arg1 type 100:103 The human immunodeficiency virus type 1 67:105 The human immunodeficiency virus type 1 (HIV-1) 67:113 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 23 theme virus 94:98 arg1 HIV-1 108:112 HIV-1 108:112 HIV-1 108:112 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 23 theme virus 94:98 arg1 machine 205:211 a conformational machine 188:211 a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state 188:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 5 24 theme Pre-fusion 807:816 arg1 gp41 818:821 Pre-fusion gp41 807:821 Pre-fusion gp41 807:821 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 2 25 theme HIV-1 384:388 arg1 surface 397:403 the HIV-1 virion surface 380:403 the HIV-1 virion surface 380:403 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 7 26 theme N-linked 1358:1365 arg1 glycan 1367:1372 N-linked glycan 1358:1372 N-linked glycan 1358:1372 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 27 theme glycan 1367:1372 arg1 tolerance 1378:1386 tolerance 1378:1386 tolerance for epitope-sequence variation 1378:1417 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 27 theme glycan 1367:1372 arg1 recognition 1343:1353 their recognition 1337:1353 their recognition of N-linked glycan 1337:1372 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 28 theme effective 1269:1277 arg1 responses 1298:1306 effective HIV-1-neutralizing responses 1269:1306 effective HIV-1-neutralizing responses 1269:1306 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 0 29 theme immune 14:19 arg1 recognition 21:31 immune recognition 14:31 immune recognition 14:31 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 2 30 theme efforts 479:485 arg1 Env 406:408 Env 406:408 Env 406:408 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 2 30 theme efforts 479:485 arg1 target 422:427 the target 418:427 the target of neutralizing antibodies 418:454 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 2 30 theme efforts 479:485 arg1 focus 462:466 a focus 460:466 a focus of vaccine efforts 460:485 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 7 31 theme chronic 1215:1221 arg1 cohorts 1223:1229 chronic cohorts 1215:1229 chronic cohorts 1215:1229 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 32 theme epitope-sequence 1392:1407 arg1 variation 1409:1417 epitope-sequence variation 1392:1417 epitope-sequence variation 1392:1417 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 33 theme HIV-1-neutralizing 1279:1296 arg1 responses 1298:1306 effective HIV-1-neutralizing responses 1269:1306 effective HIV-1-neutralizing responses 1269:1306 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 0 34 theme trimeric 36:43 arg1 Env 62:64 trimeric pre-fusion HIV-1 Env 36:64 trimeric pre-fusion HIV-1 Env 36:64 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 2 35 theme vaccine 471:477 arg1 efforts 479:485 vaccine efforts 471:485 vaccine efforts 471:485 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 2 36 theme viral 363:367 arg1 antigen 369:375 the sole viral antigen 354:375 the sole viral antigen on the HIV-1 virion surface 354:403 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 3 37 theme 3.5 Å 520:524 arg1 resolution 526:535 3.5 Å resolution 520:535 3.5 Å resolution 520:535 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 3 38 from resolution 526:535 arg1 structure 507:515 the structure 503:515 the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22 503:624 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 1 39 theme gp41 170:173 arg1 subunits 175:182 three gp120 and three gp41 subunits 148:182 subunits 175:182 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 7 40 link N-linked 1358:1365 arg1 glycan 1367:1372 N-linked glycan 1358:1372 N-linked glycan 1358:1372 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 7 41 theme N-linked 1119:1126 arg1 glycosylation 1128:1140 N-linked glycosylation 1119:1140 N-linked glycosylation 1119:1140 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 2 42 theme sole 358:361 arg1 antigen 369:375 the sole viral antigen 354:375 the sole viral antigen on the HIV-1 virion surface 354:403 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 0 43 theme HIV-1 56:60 arg1 Env 62:64 trimeric pre-fusion HIV-1 Env 36:64 trimeric pre-fusion HIV-1 Env 36:64 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 1 44 theme mature 264:269 arg1 state 282:286 a mature unliganded state 262:286 a mature unliganded state 262:286 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 5 45 theme fusion 961:966 arg1 methionine 985:994 a fusion peptide-proximal methionine 959:994 a fusion peptide-proximal methionine 959:994 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 5 46 theme carboxy-terminal 844:859 arg1 strands 861:867 amino- and carboxy-terminal strands 833:867 amino- and carboxy-terminal strands of gp120 833:876 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 0 47 theme pre-fusion 45:54 arg1 Env 62:64 trimeric pre-fusion HIV-1 Env 36:64 trimeric pre-fusion HIV-1 Env 36:64 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 1 48 theme unliganded 271:280 arg1 state 282:286 a mature unliganded state 262:286 a mature unliganded state 262:286 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 5 49 theme methionine 985:994 arg1 insertion 946:954 insertion 946:954 insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp 946:1023 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 5 50 theme peptide-proximal 968:983 arg1 methionine 985:994 a fusion peptide-proximal methionine 959:994 a fusion peptide-proximal methionine 959:994 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 7 51 theme pre-fusion 1182:1191 arg1 spike 1200:1204 the pre-fusion closed spike 1178:1204 the pre-fusion closed spike 1178:1204 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 6 52 dep clasp 1086:1090 arg1 termini 1110:1116 the termini 1106:1116 the termini 1106:1116 Spike rearrangements required for entry involve opening the clasp and expelling the termini. 25296255 6 52 dep clasp 1086:1090 arg1 the 1082:1084 the 1082:1084 the 1082:1084 Spike rearrangements required for entry involve opening the clasp and expelling the termini. 25296255 7 53 link N-linked 1119:1126 arg1 glycosylation 1128:1140 N-linked glycosylation 1119:1140 N-linked glycosylation 1119:1140 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 3 54 theme HIV-1 544:548 arg1 trimer 554:559 an HIV-1 Env trimer 541:559 an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22 541:624 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 7 55 used used 1210:1213 arg2 we 1207:1208 we 1207:1208 we 1207:1208 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 3 56 theme mature 575:580 arg1 state 589:593 a mature closed state 573:593 a mature closed state 573:593 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 3 57 theme Env 550:552 arg1 trimer 554:559 an HIV-1 Env trimer 541:559 an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22 541:624 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 5 58 theme gp120 872:876 arg1 strands 861:867 amino- and carboxy-terminal strands 833:867 amino- and carboxy-terminal strands of gp120 833:876 Pre-fusion gp41 encircles amino- and carboxy-terminal strands of gp120 with four helices that form a membrane-proximal collar, fastened by insertion of a fusion peptide-proximal methionine into a gp41-tryptophan clasp. 25296255 2 59 theme neutralizing 432:443 arg1 antibodies 445:454 neutralizing antibodies 432:454 neutralizing antibodies 432:454 As the sole viral antigen on the HIV-1 virion surface, Env is both the target of neutralizing antibodies and a focus of vaccine efforts. 25296255 3 60 theme closed 582:587 arg1 state 589:593 a mature closed state 573:593 a mature closed state 573:593 Here we report the structure at 3.5 Å resolution for an HIV-1 Env trimer captured in a mature closed state by antibodies PGT122 and 35O22. 25296255 7 61 dep used 1210:1213 arg1 distinguished 1320:1332 distinguished 1320:1332 were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation 1315:1417 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 0 62 theme Env 62:64 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 0 62 theme Env 62:64 arg1 recognition 21:31 immune recognition 14:31 immune recognition 14:31 Structure and immune recognition of trimeric pre-fusion HIV-1 Env. 25296255 4 63 theme fusion 723:728 arg1 activation 730:739 fusion activation 723:739 fusion activation 723:739 This structure reveals the pre-fusion conformation of gp41, indicates rearrangements needed for fusion activation, and defines parameters of immune evasion and immune recognition. 25296255 7 64 theme sequence-variable 1146:1162 arg1 regions 1164:1170 sequence-variable regions 1146:1170 sequence-variable regions 1146:1170 N-linked glycosylation and sequence-variable regions cover the pre-fusion closed spike; we used chronic cohorts to map the prevalence and location of effective HIV-1-neutralizing responses, which were distinguished by their recognition of N-linked glycan and tolerance for epitope-sequence variation. 25296255 1 65 theme conformational 190:203 arg1 type 100:103 The human immunodeficiency virus type 1 67:105 The human immunodeficiency virus type 1 (HIV-1) 67:113 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 25296255 1 65 theme conformational 190:203 arg1 machine 205:211 a conformational machine 188:211 a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state 188:348 The human immunodeficiency virus type 1 (HIV-1) envelope (Env) spike, comprising three gp120 and three gp41 subunits, is a conformational machine that facilitates HIV-1 entry by rearranging from a mature unliganded state, through receptor-bound intermediates, to a post-fusion state. 26316330 7 0 theme protein 1044:1050 arg1 species 1052:1058 protein species 1044:1058 protein species 1044:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 8 1 gly glycoprotein 1158:1169 arg1 glycoprotein 1158:1169 glycoprotein species 1158:1177 The new term "glycoprotein species" 1144:1178 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 8 2 theme new 1148:1150 arg1 " 1178:1178 The new term "glycoprotein species" 1144:1178 The new term "glycoprotein species" 1144:1178 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 6 3 theme glycoprotein 896:907 arg1 species 909:915 glycoprotein species 896:915 glycoprotein species 896:915 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 2 4 theme protein 276:282 arg1 species 284:290 protein species 276:290 protein species 276:290 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 5 5 theme glycoproteins 619:631 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 5 theme glycoproteins 619:631 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 5 theme glycoproteins 619:631 arg1 antibody 651:658 a therapeutical antibody 635:658 a therapeutical antibody 635:658 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 10 6 theme conditions 1386:1395 arg1 influence 1362:1370 The influence 1358:1370 The influence of production conditions for recombinant glycoproteins on glycoprotein species generation 1358:1460 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 8 7 theme species 1171:1177 arg1 " 1178:1178 The new term "glycoprotein species" 1144:1178 The new term "glycoprotein species" 1144:1178 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 5 8 theme protease 674:681 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 8 theme protease 674:681 arg1 A1AT 714:717 A1AT 714:717 A1AT 714:717 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 8 theme protease 674:681 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 8 9 dep contributions 1126:1138 arg1 defined 1184:1190 defined 1184:1190 was defined to introduce the concept of glycoforms into the field 1180:1244 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 5 10 gly glycoproteins 619:631 arg1 glycoproteins 619:631 two recombinant glycoproteins 603:631 two recombinant glycoproteins 603:631 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 6 11 from conditions 852:861 arg1 lines 824:828 the used cell lines 810:828 the used cell lines 810:828 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 0 12 theme species 104:110 arg1 generation 82:91 the generation 78:91 the generation of protein species 78:110 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 4 13 theme species 447:453 arg1 " 454:454 glycoprotein species" 434:454 glycoprotein species" 434:454 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 6 14 theme A1AT 940:943 arg1 production 945:954 recombinant A1AT production 928:954 recombinant A1AT production 928:954 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 3 15 theme structural 297:306 arg1 heterogeneity 308:320 The structural heterogeneity 293:320 The structural heterogeneity of glycoproteins 293:337 The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. 26316330 7 16 theme species 1052:1058 arg1 amount 1034:1039 the huge amount 1025:1039 the huge amount of protein species 1025:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 16 theme species 1052:1058 arg1 species 1052:1058 protein species 1044:1058 protein species 1044:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 2 17 theme chemical 214:221 arg1 compositions 223:234 versatile chemical compositions 204:234 versatile chemical compositions of proteins 204:246 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 5 18 theme alpha-1-antitrypsin 693:711 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 18 theme alpha-1-antitrypsin 693:711 arg1 A1AT 714:717 A1AT 714:717 A1AT 714:717 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 18 theme alpha-1-antitrypsin 693:711 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 4 19 theme different 486:494 arg1 fields 496:501 different fields 486:501 different fields of biology 486:512 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 0 20 theme conditions 64:73 arg1 impact 31:36 The impact 27:36 The impact of cell lines and culture conditions on the generation of protein species 27:110 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 5 21 theme glycoprotein 579:590 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 21 theme glycoprotein 579:590 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 21 theme glycoprotein 579:590 arg1 antibody 651:658 a therapeutical antibody 635:658 a therapeutical antibody 635:658 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 1 22 theme post-translational 147:164 arg1 Glycosylation 113:125 Glycosylation 113:125 Glycosylation 113:125 Glycosylation is the most complex post-translational modification. 26316330 1 22 theme post-translational 147:164 arg1 modification 166:177 the most complex post-translational modification 130:177 the most complex post-translational modification 130:177 Glycosylation is the most complex post-translational modification. 26316330 10 23 theme recombinant 1401:1411 arg1 glycoproteins 1413:1425 recombinant glycoproteins 1401:1425 recombinant glycoproteins 1401:1425 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 10 24 dep displayed 1466:1474 arg1 3 1355:1355 3 1355:1355 3 1355:1355 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 9 25 dep given 1348:1352 arg1 2 1247:1247 2 1247:1247 2 1247:1247 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 0 26 from impact 31:36 arg1 generation 82:91 the generation 78:91 the generation of protein species 78:110 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 10 27 theme species 1443:1449 arg1 generation 1451:1460 glycoprotein species generation 1430:1460 glycoprotein species generation 1430:1460 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 9 28 gly glycoproteins 1330:1342 arg1 glycoproteins 1330:1342 two particular glycoproteins 1315:1342 two particular glycoproteins 1315:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 0 29 theme Recombinant 0:10 arg1 glycoproteins 12:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 9 30 theme glycoprotein 1291:1302 arg1 species 1304:1310 potential glycoprotein species 1281:1310 potential glycoprotein species of two particular glycoproteins 1281:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 9 31 theme particular 1319:1328 arg1 glycoproteins 1330:1342 two particular glycoproteins 1315:1342 two particular glycoproteins 1315:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 6 32 theme cultivation 840:850 arg1 conditions 852:861 their cultivation conditions 834:861 their cultivation conditions 834:861 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 0 33 gly glycoproteins 12:24 arg1 glycoproteins 12:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 0 34 theme cell 41:44 arg1 lines 46:50 cell lines 41:50 cell lines 41:50 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 8 35 theme species 1101:1107 arg1 sight 1084:1088 the sight 1080:1088 the sight of protein species 1080:1107 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 0 36 dep impact 31:36 arg1 glycoproteins 12:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins 0:24 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 6 37 theme cell 819:822 arg1 lines 824:828 the used cell lines 810:828 the used cell lines 810:828 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 10 38 gly glycoprotein 1430:1441 arg1 glycoprotein 1430:1441 glycoprotein species 1430:1449 glycoprotein species generation 1430:1460 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 5 39 gly glycoprotein 579:590 arg1 glycoprotein 579:590 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 10 40 from influence 1362:1370 arg1 generation 1451:1460 glycoprotein species generation 1430:1460 glycoprotein species generation 1430:1460 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 10 41 gly glycoproteins 1413:1425 arg1 glycoproteins 1413:1425 recombinant glycoproteins 1401:1425 recombinant glycoproteins 1401:1425 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 5 42 theme therapeutical 637:649 arg1 antibody 651:658 a therapeutical antibody 635:658 a therapeutical antibody 635:658 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 42 theme therapeutical 637:649 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 6 43 theme species 909:915 arg1 number 886:891 the number 882:891 the number of glycoprotein species 882:915 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 2 44 theme species 284:290 arg1 amounts 265:271 high amounts 260:271 high amounts of protein species 260:290 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 2 44 theme species 284:290 arg1 species 284:290 protein species 276:290 protein species 276:290 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 4 45 gly glycoprotein 434:445 arg1 glycoprotein 434:445 glycoprotein species" 434:454 glycoprotein species" 434:454 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 4 46 theme new 417:419 arg1 term 421:424 a new term 415:424 a new term called "glycoprotein species" 415:454 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 5 47 theme recombinant 607:617 arg1 glycoproteins 619:631 two recombinant glycoproteins 603:631 two recombinant glycoproteins 603:631 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 8 48 theme term 1152:1155 arg1 " 1178:1178 The new term "glycoprotein species" 1144:1178 The new term "glycoprotein species" 1144:1178 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 10 49 theme production 1375:1384 arg1 conditions 1386:1395 production conditions 1375:1395 production conditions 1375:1395 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 8 50 theme glycoprotein 1158:1169 arg1 species 1171:1177 glycoprotein species 1158:1177 The new term "glycoprotein species" 1144:1178 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 8 51 theme glycoforms 1220:1229 arg1 concept 1209:1215 the concept 1205:1215 the concept of glycoforms 1205:1229 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 7 52 theme SIGNIFICANCE 968:979 arg1 glycosylation 989:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 52 theme SIGNIFICANCE 968:979 arg1 source 1014:1019 a major source 1006:1019 a major source for the huge amount of protein species 1006:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 2 53 theme proteins 239:246 arg1 compositions 223:234 versatile chemical compositions 204:234 versatile chemical compositions of proteins 204:246 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 5 54 theme human 668:672 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 54 theme human 668:672 arg1 A1AT 714:717 A1AT 714:717 A1AT 714:717 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 54 theme human 668:672 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 7 55 theme huge 1029:1032 arg1 amount 1034:1039 the huge amount 1025:1039 the huge amount of protein species 1025:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 55 theme huge 1029:1032 arg1 species 1052:1058 protein species 1044:1058 protein species 1044:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 6 56 theme recombinant 928:938 arg1 production 945:954 recombinant A1AT production 928:954 recombinant A1AT production 928:954 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 2 57 theme high 260:263 arg1 amounts 265:271 high amounts 260:271 high amounts of protein species 260:290 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 2 57 theme high 260:263 arg1 species 284:290 protein species 276:290 protein species 276:290 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 7 58 theme Protein 981:987 arg1 glycosylation 989:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 58 theme Protein 981:987 arg1 source 1014:1019 a major source 1006:1019 a major source for the huge amount of protein species 1006:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 4 59 theme glycoprotein 434:445 arg1 " 454:454 glycoprotein species" 434:454 glycoprotein species" 434:454 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 0 60 theme culture 56:62 arg1 conditions 64:73 culture conditions 56:73 culture conditions 56:73 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 7 61 theme major 1008:1012 arg1 glycosylation 989:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 61 theme major 1008:1012 arg1 source 1014:1019 a major source 1006:1019 a major source for the huge amount of protein species 1006:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 3 62 gly glycoproteins 325:337 arg1 glycoproteins 325:337 glycoproteins 325:337 glycoproteins 325:337 The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. 26316330 5 63 theme N-glycans 760:768 arg1 analysis 742:749 structural analysis 731:749 structural analysis of their N-glycans 731:768 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 1 64 theme complex 139:145 arg1 Glycosylation 113:125 Glycosylation 113:125 Glycosylation 113:125 Glycosylation is the most complex post-translational modification. 26316330 1 64 theme complex 139:145 arg1 modification 166:177 the most complex post-translational modification 130:177 the most complex post-translational modification 130:177 Glycosylation is the most complex post-translational modification. 26316330 3 65 theme glycoproteins 325:337 arg1 heterogeneity 308:320 The structural heterogeneity 293:320 The structural heterogeneity of glycoproteins 293:337 The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. 26316330 4 66 theme biology 506:512 arg1 fields 496:501 different fields 486:501 different fields of biology 486:512 We therefore introduced a new term called "glycoprotein species" to join the two concepts from different fields of biology. 26316330 6 67 gly glycoprotein 896:907 arg1 glycoprotein 896:907 glycoprotein species 896:915 glycoprotein species 896:915 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 5 68 theme species 592:598 arg1 numbers 568:574 the theoretical numbers 552:574 the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 552:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 9 69 gly glycoprotein 1291:1302 arg1 glycoprotein 1291:1302 potential glycoprotein species 1281:1310 potential glycoprotein species of two particular glycoproteins 1281:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 0 70 theme protein 96:102 arg1 species 104:110 protein species 96:110 protein species 96:110 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 9 71 theme number 1271:1276 arg1 estimation 1253:1262 An estimation 1250:1262 An estimation of the number of potential glycoprotein species of two particular glycoproteins 1250:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 7 72 theme BIOLOGICAL 957:966 arg1 glycosylation 989:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation 957:1001 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 7 72 theme BIOLOGICAL 957:966 arg1 source 1014:1019 a major source 1006:1019 a major source for the huge amount of protein species 1006:1058 BIOLOGICAL SIGNIFICANCE Protein glycosylation is a major source for the huge amount of protein species. 26316330 6 73 from variations 796:805 arg1 lines 824:828 the used cell lines 810:828 the used cell lines 810:828 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 5 74 theme structural 731:740 arg1 analysis 742:749 structural analysis 731:749 structural analysis of their N-glycans 731:768 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 9 75 theme potential 1281:1289 arg1 species 1304:1310 potential glycoprotein species 1281:1310 potential glycoprotein species of two particular glycoproteins 1281:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 5 76 theme theoretical 556:566 arg1 numbers 568:574 the theoretical numbers 552:574 the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 552:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 10 77 theme glycoprotein 1430:1441 arg1 species 1443:1449 glycoprotein species 1430:1449 glycoprotein species generation 1430:1460 3) The influence of production conditions for recombinant glycoproteins on glycoprotein species generation was displayed. 26316330 9 78 theme species 1304:1310 arg1 number 1271:1276 the number 1267:1276 the number of potential glycoprotein species of two particular glycoproteins 1267:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 3 79 theme glycoforms 379:388 arg1 definition 365:374 the definition 361:374 the definition of glycoforms 361:388 The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. 26316330 8 80 theme protein 1093:1099 arg1 species 1101:1107 protein species 1093:1107 protein species 1093:1107 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 8 81 dep defined 1184:1190 arg1 1 1141:1141 1 1141:1141 1 1141:1141 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 0 82 theme lines 46:50 arg1 impact 31:36 The impact 27:36 The impact of cell lines and culture conditions on the generation of protein species 27:110 Recombinant glycoproteins: The impact of cell lines and culture conditions on the generation of protein species. 26316330 9 83 theme glycoproteins 1330:1342 arg1 species 1304:1310 potential glycoprotein species 1281:1310 potential glycoprotein species of two particular glycoproteins 1281:1342 2) An estimation of the number of potential glycoprotein species of two particular glycoproteins was given. 26316330 2 84 theme versatile 204:212 arg1 compositions 223:234 versatile chemical compositions 204:234 versatile chemical compositions of proteins 204:246 Thus, it contributes to versatile chemical compositions of proteins, leading to high amounts of protein species. 26316330 8 85 theme following 1116:1124 arg1 contributions 1126:1138 the following contributions 1112:1138 the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field 1112:1244 This study extends the sight of protein species by the following contributions: 1) The new term "glycoprotein species" was defined to introduce the concept of glycoforms into the field. 26316330 3 86 gly heterogeneity 308:320 arg1 glycoproteins 325:337 glycoproteins 325:337 glycoproteins 325:337 The structural heterogeneity of glycoproteins was also described by the definition of glycoforms. 26316330 6 87 theme used 814:817 arg1 lines 824:828 the used cell lines 810:828 the used cell lines 810:828 Moreover, we showed that variations in the used cell lines and their cultivation conditions strongly influence the number of glycoprotein species in case of recombinant A1AT production. 26316330 5 88 dep species 592:598 arg1 inhibitor 683:691 the human protease inhibitor alpha-1-antitrypsin 664:711 the human protease inhibitor alpha-1-antitrypsin (A1AT) 664:718 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 88 dep species 592:598 arg1 antibody 651:658 a therapeutical antibody 635:658 a therapeutical antibody 635:658 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 88 dep species 592:598 arg1 species 592:598 glycoprotein species 579:598 glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - 579:720 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 26316330 5 88 dep species 592:598 arg1 A1AT 714:717 A1AT 714:717 A1AT 714:717 In this study, we further determined the theoretical numbers of glycoprotein species of two recombinant glycoproteins - a therapeutical antibody and the human protease inhibitor alpha-1-antitrypsin (A1AT) - based on structural analysis of their N-glycans. 29016103 0 0 theme Strong 80:85 arg1 Exchange 93:100 Strong Anion Exchange 80:100 Strong Anion Exchange 80:100 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 8 1 used used 1286:1289 arg2 a 1254:1254 a 1254:1254 a 1254:1254 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 6 2 theme N-glycans 774:782 arg1 removal 763:769 removal 763:769 removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC 763:837 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 1 3 theme protein 170:176 arg1 glycosylation 178:190 protein glycosylation 170:190 protein glycosylation 170:190 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 5 4 theme glycopeptides 730:742 arg1 enrichment 700:709 enrichment 700:709 enrichment of N- and O-linked glycopeptides 700:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 6 5 gly glycopeptides 816:828 arg2 glycopeptides 816:828 O-linked glycopeptides 807:828 O-linked glycopeptides 807:828 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 7 6 theme O-linked 1163:1170 arg1 glycopeptides 1172:1184 O-linked glycopeptides 1163:1184 O-linked glycopeptides 1163:1184 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 5 7 theme N- 714:715 arg1 enrichment 700:709 enrichment 700:709 enrichment of N- and O-linked glycopeptides 700:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 6 8 theme mass 892:895 arg1 spectrometry 897:908 mass spectrometry 892:908 mass spectrometry 892:908 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 1 9 theme glycosylation 178:190 arg1 Heterogeneity 153:165 Heterogeneity 153:165 Heterogeneity of protein glycosylation 153:190 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 7 10 theme intact 946:951 arg1 N- 953:954 intact N- 946:954 intact N- 946:954 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 10 11 theme cartridges 1517:1526 arg1 use 1498:1500 the use 1494:1500 the use of SAX and RAX cartridges 1494:1526 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 8 12 theme AX 1263:1264 arg1 RAX 1277:1279 RAX 1277:1279 RAX 1277:1279 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 8 12 theme AX 1263:1264 arg1 cartridge 1266:1274 AX cartridge 1263:1274 AX cartridge (RAX) 1263:1280 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 7 13 theme N- 953:954 arg1 enrichment 932:941 the enrichment 928:941 the enrichment of intact N- and O-linked glycopeptides using other chromatography methods 928:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 1 14 theme glycoproteins 280:292 arg1 structure 254:262 structure 254:262 structure 254:262 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 1 14 theme glycoproteins 280:292 arg1 function 268:275 function 268:275 function 268:275 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 0 15 theme Anion 87:91 arg1 Exchange 93:100 Strong Anion Exchange 80:100 Strong Anion Exchange 80:100 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 7 16 theme O-linked 960:967 arg1 glycopeptides 969:981 O-linked glycopeptides 960:981 O-linked glycopeptides using other chromatography methods 960:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 5 17 theme O-linked 721:728 arg1 glycopeptides 730:742 O-linked glycopeptides 721:742 O-linked glycopeptides 721:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 6 18 theme glycopeptides 875:887 arg1 identification 848:861 identification 848:861 identification of O-linked glycopeptides 848:887 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 8 19 gly glycopeptides 1214:1226 arg2 glycopeptides 1214:1226 O-linked glycopeptides 1205:1226 O-linked glycopeptides 1205:1226 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 0 20 theme Hydrophilic 106:116 arg1 Chromatography 137:150 Hydrophilic Interaction Liquid Chromatography 106:150 Hydrophilic Interaction Liquid Chromatography 106:150 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 5 21 theme enrichment 677:686 arg1 methods 688:694 three different enrichment methods 661:694 three different enrichment methods for enrichment of N- and O-linked glycopeptides 661:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 5 22 theme different 667:675 arg1 methods 688:694 three different enrichment methods 661:694 three different enrichment methods for enrichment of N- and O-linked glycopeptides 661:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 10 23 theme RAX 1513:1515 arg1 cartridges 1517:1526 SAX and RAX cartridges 1505:1526 SAX and RAX cartridges 1505:1526 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 7 24 theme glycopeptides 969:981 arg1 enrichment 932:941 the enrichment 928:941 the enrichment of intact N- and O-linked glycopeptides using other chromatography methods 928:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 25 contain containing 1050:1059 arg2 materials 1061:1069 materials 1061:1069 materials for strong anion exchange (SAX) chromatography 1061:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 25 contain containing 1050:1059 arg1 cartridges 1039:1048 cartridges 1039:1048 cartridges containing materials for strong anion exchange (SAX) chromatography 1039:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 10 26 theme broad 1544:1548 arg1 applications 1550:1561 broad applications 1544:1561 broad applications of identifications 1544:1580 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 3 27 gly glycopeptides 540:552 arg2 glycopeptides 540:552 O-linked glycopeptides 531:552 O-linked glycopeptides 531:552 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 9 28 theme labeled 1320:1326 arg1 glycopeptides 1328:1340 isobaric tag labeled glycopeptides 1307:1340 isobaric tag labeled glycopeptides after C18 desalting 1307:1360 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 1 29 theme great 198:202 arg1 challenges 204:213 great challenges 198:213 great challenges 198:213 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 6 30 link O-linked 807:814 arg1 glycopeptides 816:828 O-linked glycopeptides 807:828 O-linked glycopeptides 807:828 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 7 31 theme glycopeptides 1172:1184 arg1 identification 1138:1151 identification 1138:1151 identification 1138:1151 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 31 theme glycopeptides 1172:1184 arg1 yield 1128:1132 yield 1128:1132 yield 1128:1132 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 10 32 theme SAX 1505:1507 arg1 cartridges 1517:1526 SAX and RAX cartridges 1505:1526 SAX and RAX cartridges 1505:1526 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 3 33 used used 502:505 arg2 HILIC 477:481 HILIC 477:481 HILIC 477:481 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 3 33 used used 502:505 arg2 chromatography 461:474 hydrophilic interaction chromatography 437:474 hydrophilic interaction chromatography (HILIC) 437:482 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 10 34 gly glycoproteins 1602:1614 arg1 glycoproteins 1602:1614 glycoproteins 1602:1614 glycoproteins 1602:1614 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 9 35 gly glycopeptides 1328:1340 arg2 glycopeptides 1328:1340 isobaric tag labeled glycopeptides 1307:1340 isobaric tag labeled glycopeptides after C18 desalting 1307:1360 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 10 36 theme identifications 1566:1580 arg1 quantitation 1586:1597 quantitation 1586:1597 quantitation of glycoproteins 1586:1614 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 10 36 theme identifications 1566:1580 arg1 applications 1550:1561 broad applications 1544:1561 broad applications of identifications 1544:1580 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 4 37 theme labeled 605:611 arg1 glycopeptides 613:625 isobarically labeled glycopeptides 592:625 isobarically labeled glycopeptides 592:625 However, its effectiveness to enrich isobarically labeled glycopeptides remains unclear. 29016103 9 38 theme quantitative 1441:1452 arg1 glycoproteomics 1454:1468 quantitative glycoproteomics 1441:1468 quantitative glycoproteomics 1441:1468 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 2 39 theme specific 343:350 arg1 enrichment 352:361 efficient and specific enrichment 329:361 efficient and specific enrichment of intact glycopeptides for identification and quantitation 329:421 Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. 29016103 7 40 gly glycopeptides 1172:1184 arg2 glycopeptides 1172:1184 O-linked glycopeptides 1163:1184 O-linked glycopeptides 1163:1184 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 0 41 theme Methods 25:31 arg1 Comparison 0:9 Comparison 0:9 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides 0:72 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 6 42 theme O-linked 807:814 arg1 glycopeptides 816:828 O-linked glycopeptides 807:828 O-linked glycopeptides 807:828 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 0 43 theme Liquid 130:135 arg1 Chromatography 137:150 Hydrophilic Interaction Liquid Chromatography 106:150 Hydrophilic Interaction Liquid Chromatography 106:150 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 1 44 gly glycoproteins 280:292 arg1 glycoproteins 280:292 glycoproteins 280:292 glycoproteins 280:292 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 0 45 theme Enrichment 14:23 arg1 Methods 25:31 Enrichment Methods 14:31 Enrichment Methods for Intact N- and O-Linked Glycopeptides 14:72 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 0 46 theme Interaction 118:128 arg1 Chromatography 137:150 Hydrophilic Interaction Liquid Chromatography 106:150 Hydrophilic Interaction Liquid Chromatography 106:150 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 9 47 theme isobaric 1307:1314 arg1 glycopeptides 1328:1340 isobaric tag labeled glycopeptides 1307:1340 isobaric tag labeled glycopeptides after C18 desalting 1307:1360 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 2 48 theme efficient 329:337 arg1 enrichment 352:361 efficient and specific enrichment 329:361 efficient and specific enrichment of intact glycopeptides for identification and quantitation 329:421 Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. 29016103 8 49 link O-linked 1205:1212 arg1 glycopeptides 1214:1226 O-linked glycopeptides 1205:1226 O-linked glycopeptides 1205:1226 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 5 50 link O-linked 721:728 arg1 glycopeptides 730:742 O-linked glycopeptides 721:742 O-linked glycopeptides 721:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 0 51 theme Intact 37:42 arg1 N- 44:45 Intact N- 37:45 Intact N- 37:45 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 3 52 theme O-linked 531:538 arg1 glycopeptides 540:552 O-linked glycopeptides 531:552 O-linked glycopeptides 531:552 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 8 53 theme O-linked 1205:1212 arg1 glycopeptides 1214:1226 O-linked glycopeptides 1205:1226 O-linked glycopeptides 1205:1226 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 9 54 theme tag 1316:1318 arg1 glycopeptides 1328:1340 isobaric tag labeled glycopeptides 1307:1340 isobaric tag labeled glycopeptides after C18 desalting 1307:1360 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 7 55 link O-linked 960:967 arg1 glycopeptides 969:981 O-linked glycopeptides 960:981 O-linked glycopeptides using other chromatography methods 960:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 56 theme N- 1156:1157 arg1 identification 1138:1151 identification 1138:1151 identification 1138:1151 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 56 theme N- 1156:1157 arg1 yield 1128:1132 yield 1128:1132 yield 1128:1132 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 57 link O-linked 1163:1170 arg1 glycopeptides 1172:1184 O-linked glycopeptides 1163:1184 O-linked glycopeptides 1163:1184 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 3 58 theme hydrophilic 437:447 arg1 HILIC 477:481 HILIC 477:481 HILIC 477:481 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 3 58 theme hydrophilic 437:447 arg1 chromatography 461:474 hydrophilic interaction chromatography 437:474 hydrophilic interaction chromatography (HILIC) 437:482 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 2 59 gly glycopeptides 373:385 arg2 glycopeptides 373:385 intact glycopeptides 366:385 intact glycopeptides 366:385 Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. 29016103 6 60 link O-linked 866:873 arg1 glycopeptides 875:887 O-linked glycopeptides 866:887 O-linked glycopeptides 866:887 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 6 61 theme glycopeptides 816:828 arg1 enrichment 793:802 enrichment 793:802 enrichment of O-linked glycopeptides 793:828 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 3 62 theme interaction 449:459 arg1 HILIC 477:481 HILIC 477:481 HILIC 477:481 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 3 62 theme interaction 449:459 arg1 chromatography 461:474 hydrophilic interaction chromatography 437:474 hydrophilic interaction chromatography (HILIC) 437:482 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 7 63 theme strong 1075:1080 arg1 chromatography 1103:1116 strong anion exchange (SAX) chromatography 1075:1116 strong anion exchange (SAX) chromatography 1075:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 1 64 gly Heterogeneity 153:165 arg1 glycosylation 178:190 protein glycosylation 170:190 protein glycosylation 170:190 Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. 29016103 8 65 dep a 1254:1254 arg1 Retain 1256:1261 Retain 1256:1261 Retain AX cartridge (RAX) 1256:1280 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 29016103 4 66 gly glycopeptides 613:625 arg2 glycopeptides 613:625 isobarically labeled glycopeptides 592:625 isobarically labeled glycopeptides 592:625 However, its effectiveness to enrich isobarically labeled glycopeptides remains unclear. 29016103 2 67 theme glycopeptides 373:385 arg1 enrichment 352:361 efficient and specific enrichment 329:361 efficient and specific enrichment of intact glycopeptides for identification and quantitation 329:421 Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. 29016103 7 68 theme other 989:993 arg1 methods 1010:1016 other chromatography methods 989:1016 other chromatography methods 989:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 6 69 gly glycopeptides 875:887 arg2 glycopeptides 875:887 O-linked glycopeptides 866:887 O-linked glycopeptides 866:887 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 10 70 theme glycoproteins 1602:1614 arg1 quantitation 1586:1597 quantitation 1586:1597 quantitation of glycoproteins 1586:1614 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 10 70 theme glycoproteins 1602:1614 arg1 applications 1550:1561 broad applications 1544:1561 broad applications of identifications 1544:1580 It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins. 29016103 9 71 theme C18 1348:1350 arg1 desalting 1352:1360 C18 desalting 1348:1360 C18 desalting 1348:1360 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 7 72 theme SAX 1098:1100 arg1 chromatography 1103:1116 strong anion exchange (SAX) chromatography 1075:1116 strong anion exchange (SAX) chromatography 1075:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 2 73 theme intact 366:371 arg1 glycopeptides 373:385 intact glycopeptides 366:385 intact glycopeptides 366:385 Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. 29016103 7 74 theme chromatography 995:1008 arg1 methods 1010:1016 other chromatography methods 989:1016 other chromatography methods 989:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 3 75 link O-linked 531:538 arg1 glycopeptides 540:552 O-linked glycopeptides 531:552 O-linked glycopeptides 531:552 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 7 76 theme anion 1082:1086 arg1 chromatography 1103:1116 strong anion exchange (SAX) chromatography 1075:1116 strong anion exchange (SAX) chromatography 1075:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 9 77 dep SAX 1391:1393 arg1 cartridges 1403:1412 cartridges 1403:1412 cartridges 1403:1412 In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. 29016103 6 78 theme O-linked 866:873 arg1 glycopeptides 875:887 O-linked glycopeptides 866:887 O-linked glycopeptides 866:887 It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. 29016103 3 79 theme intact 517:522 arg1 N- 524:525 intact N- 517:525 intact N- 517:525 To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. 29016103 7 80 gly glycopeptides 969:981 arg2 glycopeptides 969:981 O-linked glycopeptides 960:981 O-linked glycopeptides using other chromatography methods 960:1016 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 7 81 theme exchange 1088:1095 arg1 chromatography 1103:1116 strong anion exchange (SAX) chromatography 1075:1116 strong anion exchange (SAX) chromatography 1075:1116 We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. 29016103 0 82 theme O-Linked 51:58 arg1 Glycopeptides 60:72 O-Linked Glycopeptides 51:72 O-Linked Glycopeptides 51:72 Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography. 29016103 5 83 gly glycopeptides 730:742 arg2 glycopeptides 730:742 O-linked glycopeptides 721:742 O-linked glycopeptides 721:742 Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. 29016103 8 84 theme glycopeptides 1214:1226 arg1 enrichment 1191:1200 The enrichment 1187:1200 The enrichment of O-linked glycopeptides 1187:1226 The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. 27900782 7 0 theme residue-specific 808:823 arg1 information 834:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information is stored internally, permitting glycoengineering and design. 27900782 3 1 theme Rosetta 353:359 arg1 modeling 361:368 the Rosetta modeling 349:368 the Rosetta modeling 349:368 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 4 2 theme scoring 545:551 arg1 challenges 518:527 challenges 518:527 challenges of sampling and scoring 518:551 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 1 3 theme saccharide 160:169 arg1 structures 190:199 glycoconjugate structures 175:199 glycoconjugate structures 175:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 3 theme saccharide 160:169 arg1 variety 149:155 a wide variety 142:155 a wide variety of saccharide and glycoconjugate structures 142:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 3 theme saccharide 160:169 arg1 saccharide 160:169 saccharide 160:169 saccharide 160:169 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 8 4 theme Carbohydrate-specific 908:928 arg1 applications 930:941 Carbohydrate-specific applications 908:941 Carbohydrate-specific applications described herein 908:958 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 4 5 theme ring 633:636 arg1 forms 638:642 ring forms 633:642 ring forms 633:642 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 0 6 theme Residue-centric 0:14 arg1 modeling 16:23 Residue-centric modeling 0:23 Residue-centric modeling 0:23 Residue-centric modeling and design of saccharide and glycoconjugate structures. 27900782 1 7 theme structures 190:199 arg1 structures 190:199 glycoconjugate structures 175:199 glycoconjugate structures 175:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 7 theme structures 190:199 arg1 variety 149:155 a wide variety 142:155 a wide variety of saccharide and glycoconjugate structures 142:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 7 theme structures 190:199 arg1 saccharide 160:169 saccharide 160:169 saccharide 160:169 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 8 theme RosettaCarbohydrate 85:103 arg1 tool 124:127 a new tool 118:127 a new tool for modeling a wide variety of saccharide and glycoconjugate structures 118:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 8 theme RosettaCarbohydrate 85:103 arg1 framework 105:113 The RosettaCarbohydrate framework 81:113 The RosettaCarbohydrate framework 81:113 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 9 theme glycoconjugate 175:188 arg1 structures 190:199 glycoconjugate structures 175:199 glycoconjugate structures 175:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 9 10 theme glyco-ligand 1173:1184 arg1 binding 1186:1192 glyco-ligand binding 1173:1192 glyco-ligand binding 1173:1192 Benchmarking data are presented and compared to other studies, demonstrating Rosetta's ability to predict glyco-ligand binding. 27900782 3 11 theme carbohydrate 438:449 arg1 molecules 451:459 carbohydrate molecules 438:459 carbohydrate molecules 438:459 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 11 theme carbohydrate 438:449 arg1 branching 472:480 branching 472:480 branching 472:480 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 11 theme carbohydrate 438:449 arg1 modifications 492:504 sugar modifications 486:504 sugar modifications 486:504 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 12 theme vast 407:410 arg1 complexity 412:421 the vast complexity 403:421 the vast complexity 403:421 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 10 13 theme available 1227:1235 arg1 tools 1221:1225 the tools 1217:1225 the tools available to glycoscientists and engineers 1217:1268 The framework expands the tools available to glycoscientists and engineers. 27900782 8 14 theme virtual 968:974 arg1 glycosylation 976:988 virtual glycosylation 968:988 virtual glycosylation 968:988 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 3 15 theme molecules 451:459 arg1 branching 472:480 branching 472:480 branching 472:480 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 15 theme molecules 451:459 arg1 variety 427:433 variety 427:433 variety 427:433 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 15 theme molecules 451:459 arg1 molecules 451:459 carbohydrate molecules 438:459 carbohydrate molecules 438:459 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 15 theme molecules 451:459 arg1 complexity 412:421 the vast complexity 403:421 the vast complexity 403:421 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 3 15 theme molecules 451:459 arg1 modifications 492:504 sugar modifications 486:504 sugar modifications 486:504 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 4 16 theme glycosidic 585:594 arg1 bonds 596:600 glycosidic bonds 585:600 glycosidic bonds 585:600 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 6 17 theme file 783:786 arg1 formats 788:794 gws) file formats 778:794 gws) file formats. 778:795 gws) file formats. 27900782 3 18 theme sugar 486:490 arg1 modifications 492:504 sugar modifications 486:504 sugar modifications 486:504 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 9 19 theme Benchmarking 1067:1078 arg1 data 1080:1083 Benchmarking data 1067:1083 Benchmarking data 1067:1083 Benchmarking data are presented and compared to other studies, demonstrating Rosetta's ability to predict glyco-ligand binding. 27900782 4 20 theme side-chain 603:612 arg1 conformations 614:626 side-chain conformations 603:626 side-chain conformations 603:626 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 5 21 theme standard 735:742 arg1 PDB 744:746 standard PDB 735:746 standard PDB 735:746 Rosetta can work with standard PDB, GLYCAM, and GlycoWorkbench (. 27900782 1 22 theme new 120:122 arg1 tool 124:127 a new tool 118:127 a new tool for modeling a wide variety of saccharide and glycoconjugate structures 118:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 22 theme new 120:122 arg1 framework 105:113 The RosettaCarbohydrate framework 81:113 The RosettaCarbohydrate framework 81:113 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 0 23 theme saccharide 39:48 arg1 structures 69:78 saccharide and glycoconjugate structures 39:78 saccharide and glycoconjugate structures 39:78 Residue-centric modeling and design of saccharide and glycoconjugate structures. 27900782 7 24 theme chemical 825:832 arg1 information 834:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information is stored internally, permitting glycoengineering and design. 27900782 3 25 dep modeling 361:368 arg1 suite 381:385 suite 381:385 suite 381:385 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 2 26 theme framework 247:255 arg1 development 228:238 the development 224:238 the development of the framework 224:255 This report describes the development of the framework and highlights its applications. 27900782 4 27 theme scoring 695:701 arg1 function 703:710 its scoring function 691:710 its scoring function 691:710 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 4 28 theme glycan-specific 663:677 arg1 term 679:682 a glycan-specific term 661:682 a glycan-specific term 661:682 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 0 29 theme glycoconjugate 54:67 arg1 structures 69:78 saccharide and glycoconjugate structures 39:78 saccharide and glycoconjugate structures 39:78 Residue-centric modeling and design of saccharide and glycoconjugate structures. 27900782 7 30 theme Saccharide 797:806 arg1 information 834:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information 797:844 Saccharide residue-specific chemical information is stored internally, permitting glycoengineering and design. 27900782 6 31 theme gws 778:780 arg1 formats 788:794 gws) file formats 778:794 gws) file formats. 778:795 gws) file formats. 27900782 11 32 dep © 1271:1271 arg1 Inc. 1297:1300 Inc. 1297:1300 Inc. 1297:1300 © 2016 Wiley Periodicals, Inc. 27900782 8 33 gly glycosylation 976:988 arg1 carbohydrates 1008:1020 carbohydrates 1008:1020 carbohydrates 1008:1020 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 33 gly glycosylation 976:988 arg1 glyco-ligands 1038:1050 glyco-ligands 1038:1050 glyco-ligands 1038:1050 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 34 theme carbohydrates 1008:1020 arg1 docking 1027:1033 docking 1027:1033 docking of glyco-ligands to antibodies 1027:1064 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 34 theme carbohydrates 1008:1020 arg1 loop-modeling 991:1003 loop-modeling 991:1003 loop-modeling of carbohydrates 991:1020 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 34 theme carbohydrates 1008:1020 arg1 glycosylation 976:988 virtual glycosylation 968:988 virtual glycosylation 968:988 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 3 35 theme established 320:330 arg1 protocols 332:340 established protocols 320:340 established protocols 320:340 The framework integrates with established protocols within the Rosetta modeling and design suite, and it handles the vast complexity and variety of carbohydrate molecules, including branching and sugar modifications. 27900782 9 36 theme other 1115:1119 arg1 studies 1121:1127 other studies 1115:1127 other studies 1115:1127 Benchmarking data are presented and compared to other studies, demonstrating Rosetta's ability to predict glyco-ligand binding. 27900782 1 37 theme wide 144:147 arg1 structures 190:199 glycoconjugate structures 175:199 glycoconjugate structures 175:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 37 theme wide 144:147 arg1 variety 149:155 a wide variety 142:155 a wide variety of saccharide and glycoconjugate structures 142:199 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 1 37 theme wide 144:147 arg1 saccharide 160:169 saccharide 160:169 saccharide 160:169 The RosettaCarbohydrate framework is a new tool for modeling a wide variety of saccharide and glycoconjugate structures. 27900782 0 38 theme structures 69:78 arg1 design 29:34 design 29:34 design 29:34 Residue-centric modeling and design of saccharide and glycoconjugate structures. 27900782 0 38 theme structures 69:78 arg1 modeling 16:23 Residue-centric modeling 0:23 Residue-centric modeling 0:23 Residue-centric modeling and design of saccharide and glycoconjugate structures. 27900782 4 39 theme sampling 532:539 arg1 challenges 518:527 challenges 518:527 challenges of sampling and scoring 518:551 To address challenges of sampling and scoring, RosettaCarbohydrate can sample glycosidic bonds, side-chain conformations, and ring forms, and it utilizes a glycan-specific term within its scoring function. 27900782 8 40 theme glyco-ligands 1038:1050 arg1 docking 1027:1033 docking 1027:1033 docking of glyco-ligands to antibodies 1027:1064 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 40 theme glyco-ligands 1038:1050 arg1 loop-modeling 991:1003 loop-modeling 991:1003 loop-modeling of carbohydrates 991:1020 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27900782 8 40 theme glyco-ligands 1038:1050 arg1 glycosylation 976:988 virtual glycosylation 968:988 virtual glycosylation 968:988 Carbohydrate-specific applications described herein include virtual glycosylation, loop-modeling of carbohydrates, and docking of glyco-ligands to antibodies. 27269286 4 0 dep enzyme 664:669 arg1 β1,3-N-acetylglucosaminyltransferase 671:706 β1,3-N-acetylglucosaminyltransferase 671:706 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 3 1 theme branching 333:341 arg1 deficiency 343:352 N-glycan branching deficiency 324:352 N-glycan branching deficiency 324:352 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 8 2 theme N-glycans 1224:1232 arg1 equivalency 1185:1195 the functional equivalency 1170:1195 the functional equivalency of structurally dissimilar N-glycans 1170:1232 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 5 3 theme LacNAc 737:742 arg1 content 744:750 Marginally reducing LacNAc content 717:750 Marginally reducing LacNAc content by limiting N-glycans to three branches 717:790 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 3 4 theme galectin-glycoprotein 423:443 arg1 interactions 445:456 galectin-glycoprotein interactions 423:456 galectin-glycoprotein interactions 423:456 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 6 5 theme poly-LacNAc 931:941 arg1 extension 943:951 new poly-LacNAc extension 927:951 new poly-LacNAc extension by B3GNT 927:960 Rather, new poly-LacNAc extension by B3GNT maintains galectin binding and immune homeostasis. 27269286 5 6 theme T-cell 803:808 arg1 hyperactivity 810:822 T-cell hyperactivity 803:822 T-cell hyperactivity 803:822 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 8 7 theme functional 1174:1183 arg1 equivalency 1185:1195 the functional equivalency 1170:1195 the functional equivalency of structurally dissimilar N-glycans 1170:1232 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 5 8 theme restricting 854:864 arg1 branching 866:874 yet further restricting branching 842:874 yet further restricting branching 842:874 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 7 9 theme inter-cisternal 1122:1136 arg1 tubules 1138:1144 inter-cisternal tubules 1122:1144 inter-cisternal tubules 1122:1144 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 7 10 from redistribution 1051:1064 arg1 medial 1096:1101 medial 1096:1101 medial 1096:1101 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 2 11 theme cell 195:198 arg1 function 200:207 Mammalian cell function 185:207 Mammalian cell function 185:207 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 4 12 theme N-acetyllactosamine 499:517 arg1 units 528:532 N-acetyllactosamine (LacNAc) units 499:532 N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 499:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 1 13 theme Essential 88:96 arg1 systems 109:115 Essential biological systems 88:115 Essential biological systems 88:115 Essential biological systems employ self-correcting mechanisms to maintain cellular homeostasis. 27269286 2 14 theme Mammalian 185:193 arg1 function 200:207 Mammalian cell function 185:207 Mammalian cell function 185:207 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 8 15 theme self-correcting 1248:1262 arg1 feature 1264:1270 a self-correcting feature 1246:1270 a self-correcting feature of the Golgi that sustains cellular homeostasis 1246:1318 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 1 16 theme biological 98:107 arg1 systems 109:115 Essential biological systems 88:115 Essential biological systems 88:115 Essential biological systems employ self-correcting mechanisms to maintain cellular homeostasis. 27269286 4 17 theme medial-Golgi 584:595 arg1 enzymes 607:613 the medial-Golgi branching enzymes 580:613 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 5 18 theme hyperactive 900:910 arg1 state 912:916 a more hyperactive state 893:916 a more hyperactive state 893:916 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 7 19 theme Poly-LacNAc 1013:1023 arg1 extension 1025:1033 Poly-LacNAc extension 1013:1033 Poly-LacNAc extension 1013:1033 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 0 20 theme Golgi 0:4 arg1 self-correction 6:20 Golgi self-correction 0:20 Golgi self-correction 0:20 Golgi self-correction generates bioequivalent glycans to preserve cellular homeostasis. 27269286 6 21 theme new 927:929 arg1 extension 943:951 new poly-LacNAc extension 927:951 new poly-LacNAc extension by B3GNT 927:960 Rather, new poly-LacNAc extension by B3GNT maintains galectin binding and immune homeostasis. 27269286 0 22 theme bioequivalent 32:44 arg1 glycans 46:52 bioequivalent glycans 32:52 bioequivalent glycans 32:52 Golgi self-correction generates bioequivalent glycans to preserve cellular homeostasis. 27269286 8 23 theme dissimilar 1213:1222 arg1 N-glycans 1224:1232 structurally dissimilar N-glycans 1200:1232 structurally dissimilar N-glycans 1200:1232 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 3 24 theme N-glycan 324:331 arg1 deficiency 343:352 N-glycan branching deficiency 324:352 N-glycan branching deficiency 324:352 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 5 25 theme reducing 728:735 arg1 content 744:750 Marginally reducing LacNAc content 717:750 Marginally reducing LacNAc content by limiting N-glycans to three branches 717:790 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 1 26 theme self-correcting 124:138 arg1 mechanisms 140:149 self-correcting mechanisms 124:149 self-correcting mechanisms 124:149 Essential biological systems employ self-correcting mechanisms to maintain cellular homeostasis. 27269286 6 27 theme galectin 972:979 arg1 binding 981:987 galectin binding 972:987 galectin binding 972:987 Rather, new poly-LacNAc extension by B3GNT maintains galectin binding and immune homeostasis. 27269286 3 28 theme cellular 462:469 arg1 homeostasis 471:481 cellular homeostasis 462:481 cellular homeostasis 462:481 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 3 29 gly galectin-glycoprotein 423:443 arg1 galectin-glycoprotein 423:443 galectin-glycoprotein 423:443 galectin-glycoprotein 423:443 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 8 30 theme cellular 1299:1306 arg1 homeostasis 1308:1318 cellular homeostasis 1299:1318 cellular homeostasis 1299:1318 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 2 31 theme branched 284:291 arg1 N-glycans 293:301 branched N-glycans 284:301 branched N-glycans 284:301 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 4 32 theme trans-Golgi 630:640 arg1 enzyme 664:669 the trans-Golgi poly-LacNAc extension enzyme 626:669 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 4 32 theme trans-Golgi 630:640 arg1 B3GNT 709:713 B3GNT 709:713 B3GNT 709:713 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 0 33 theme cellular 66:73 arg1 homeostasis 75:85 cellular homeostasis 66:85 cellular homeostasis 66:85 Golgi self-correction generates bioequivalent glycans to preserve cellular homeostasis. 27269286 2 34 theme galectins 269:277 arg1 interaction 241:251 the interaction 237:251 the interaction of cell surface galectins with branched N-glycans 237:301 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 4 35 theme branching 597:605 arg1 enzymes 607:613 the medial-Golgi branching enzymes 580:613 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 7 36 from medial 1096:1101 arg1 redistribution 1051:1064 redistribution 1051:1064 redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules 1051:1144 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 7 36 from medial 1096:1101 arg1 UDP-GlcNAc 1076:1085 unused UDP-GlcNAc 1069:1085 unused UDP-GlcNAc from the medial 1069:1101 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 6 37 theme immune 993:998 arg1 homeostasis 1000:1010 immune homeostasis 993:1010 immune homeostasis 993:1010 Rather, new poly-LacNAc extension by B3GNT maintains galectin binding and immune homeostasis. 27269286 4 38 theme extension 654:662 arg1 enzyme 664:669 the trans-Golgi poly-LacNAc extension enzyme 626:669 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 4 38 theme extension 654:662 arg1 B3GNT 709:713 B3GNT 709:713 B3GNT 709:713 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 5 39 theme further 846:852 arg1 branching 866:874 yet further restricting branching 842:874 yet further restricting branching 842:874 Marginally reducing LacNAc content by limiting N-glycans to three branches results in T-cell hyperactivity and autoimmunity; yet further restricting branching does not produce a more hyperactive state. 27269286 2 40 theme cell 256:259 arg1 galectins 269:277 cell surface galectins 256:277 cell surface galectins 256:277 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 2 41 theme surface 261:267 arg1 galectins 269:277 cell surface galectins 256:277 cell surface galectins 256:277 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 4 42 theme poly-LacNAc 642:652 arg1 enzyme 664:669 the trans-Golgi poly-LacNAc extension enzyme 626:669 the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 580:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 4 42 theme poly-LacNAc 642:652 arg1 B3GNT 709:713 B3GNT 709:713 B3GNT 709:713 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 4 43 theme LacNAc 520:525 arg1 units 528:532 N-acetyllactosamine (LacNAc) units 499:532 N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT) 499:714 Galectins bind N-acetyllactosamine (LacNAc) units within N-glycans initiated from UDP-GlcNAc by the medial-Golgi branching enzymes as well as the trans-Golgi poly-LacNAc extension enzyme β1,3-N-acetylglucosaminyltransferase (B3GNT). 27269286 1 44 theme cellular 163:170 arg1 homeostasis 172:182 cellular homeostasis 163:182 cellular homeostasis 163:182 Essential biological systems employ self-correcting mechanisms to maintain cellular homeostasis. 27269286 8 45 theme Golgi 1279:1283 arg1 feature 1264:1270 a self-correcting feature 1246:1270 a self-correcting feature of the Golgi that sustains cellular homeostasis 1246:1318 These data demonstrate the functional equivalency of structurally dissimilar N-glycans and suggest a self-correcting feature of the Golgi that sustains cellular homeostasis. 27269286 7 46 theme unused 1069:1074 arg1 UDP-GlcNAc 1076:1085 unused UDP-GlcNAc 1069:1085 unused UDP-GlcNAc from the medial 1069:1101 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 27269286 3 47 theme bioequivalent 385:397 arg1 N-glycans 399:407 bioequivalent N-glycans 385:407 bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis 385:481 Here we report that N-glycan branching deficiency triggers the Golgi to generate bioequivalent N-glycans that preserve galectin-glycoprotein interactions and cellular homeostasis. 27269286 2 48 with interaction 241:251 arg1 N-glycans 293:301 branched N-glycans 284:301 branched N-glycans 284:301 Mammalian cell function is dynamically regulated by the interaction of cell surface galectins with branched N-glycans. 27269286 7 49 theme UDP-GlcNAc 1076:1085 arg1 redistribution 1051:1064 redistribution 1051:1064 redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules 1051:1144 Poly-LacNAc extension is triggered by redistribution of unused UDP-GlcNAc from the medial to trans-Golgi via inter-cisternal tubules. 28960760 0 0 theme Catalytic 93:101 arg1 Mechanism 103:111 the Catalytic Mechanism 89:111 the Catalytic Mechanism of Retaining Glycosyltransferases 89:145 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 4 1 theme divalent 963:970 arg1 cofactor 979:986 the divalent cation cofactor 959:986 the divalent cation cofactor 959:986 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 2 theme catalytic 420:428 arg1 mechanism 430:438 the catalytic mechanism 416:438 the catalytic mechanism of retaining GTs 416:455 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 2 theme catalytic 420:428 arg1 topic 462:466 a topic 460:466 a topic 460:466 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 1 3 theme living 258:263 arg1 organisms 265:273 all living organisms 254:273 all living organisms 254:273 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 1 4 theme key 181:183 arg1 family 185:190 a key family 179:190 a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms 179:273 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 1 4 theme key 181:183 arg1 Glycosyltransferases 148:167 Glycosyltransferases 148:167 Glycosyltransferases (GTs) 148:173 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 4 5 theme acceptor 927:934 arg1 lactose 946:952 the acceptor substrate lactose 923:952 the acceptor substrate lactose 923:952 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 5 6 theme conserved 1125:1133 arg1 mechanism 1152:1160 a conserved common catalytic mechanism 1123:1160 a conserved common catalytic mechanism among retaining GTs 1123:1180 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 3 7 theme putative 534:541 arg1 residue 556:562 a putative nucleophilic residue 532:562 a putative nucleophilic residue in the active site 532:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 5 8 theme reaction 1089:1096 arg1 occurrence 1033:1042 the occurrence 1029:1042 the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT 1029:1107 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 3 9 theme nucleophilic 543:554 arg1 residue 556:562 a putative nucleophilic residue 532:562 a putative nucleophilic residue in the active site 532:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 10 theme glycosyltransferase 716:734 arg1 α3GalT 765:770 α3GalT 765:770 α3GalT 765:770 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 10 theme glycosyltransferase 716:734 arg1 α-1,3-galactosyltransferase 736:762 the retaining glycosyltransferase α-1,3-galactosyltransferase 702:762 the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) 702:771 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 0 11 theme Glycosyltransferases 126:145 arg1 Mechanism 103:111 the Catalytic Mechanism 89:111 the Catalytic Mechanism of Retaining Glycosyltransferases 89:145 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 4 12 theme retaining 706:714 arg1 α3GalT 765:770 α3GalT 765:770 α3GalT 765:770 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 12 theme retaining 706:714 arg1 α-1,3-galactosyltransferase 736:762 the retaining glycosyltransferase α-1,3-galactosyltransferase 702:762 the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) 702:771 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 13 from Bos 778:780 arg1 complexes 689:697 native ternary complexes 674:697 native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal 674:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 13 from Bos 778:780 arg1 lactose 946:952 the acceptor substrate lactose 923:952 the acceptor substrate lactose 923:952 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 13 from Bos 778:780 arg1 cofactor 979:986 the divalent cation cofactor 959:986 the divalent cation cofactor 959:986 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 2 14 theme anomeric 383:390 arg1 configuration 392:404 the anomeric configuration 379:404 the anomeric configuration 379:404 The reaction involves the transfer of a glycosyl moiety and can proceed with retention or inversion of the anomeric configuration. 28960760 5 15 theme i-type 1082:1087 arg1 reaction 1089:1096 a front-side substrate-assisted SN i-type reaction 1047:1096 a front-side substrate-assisted SN i-type reaction for α3GalT 1047:1107 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 0 16 theme Retaining 116:124 arg1 Glycosyltransferases 126:145 Retaining Glycosyltransferases 116:145 Retaining Glycosyltransferases 116:145 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 2 17 theme configuration 392:404 arg1 inversion 366:374 inversion 366:374 inversion 366:374 The reaction involves the transfer of a glycosyl moiety and can proceed with retention or inversion of the anomeric configuration. 28960760 2 17 theme configuration 392:404 arg1 retention 353:361 retention 353:361 retention 353:361 The reaction involves the transfer of a glycosyl moiety and can proceed with retention or inversion of the anomeric configuration. 28960760 4 18 theme productive 849:858 arg1 mode 860:863 a productive mode 847:863 a productive mode for catalysis 847:877 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 2 19 theme moiety 325:330 arg1 transfer 302:309 the transfer 298:309 the transfer of a glycosyl moiety 298:330 The reaction involves the transfer of a glycosyl moiety and can proceed with retention or inversion of the anomeric configuration. 28960760 1 20 theme enzymes 195:201 arg1 family 185:190 a key family 179:190 a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms 179:273 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 1 20 theme enzymes 195:201 arg1 Glycosyltransferases 148:167 Glycosyltransferases 148:167 Glycosyltransferases (GTs) 148:173 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 0 21 theme Structural 0:9 arg1 Snapshots 11:19 Structural Snapshots 0:19 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 0:146 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 5 22 theme retaining 1168:1176 arg1 GTs 1178:1180 retaining GTs 1168:1180 retaining GTs 1168:1180 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 2 23 theme glycosyl 316:323 arg1 moiety 325:330 a glycosyl moiety 314:330 a glycosyl moiety 314:330 The reaction involves the transfer of a glycosyl moiety and can proceed with retention or inversion of the anomeric configuration. 28960760 0 24 theme α-1,3-Galactosyltransferase 24:50 arg1 Snapshots 11:19 Structural Snapshots 0:19 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 0:146 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 4 25 theme donor 908:912 arg1 UDP-Gal 914:920 its sugar donor UDP-Gal 898:920 its sugar donor UDP-Gal 898:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 26 theme active 571:576 arg1 site 578:581 the active site 567:581 the active site 567:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 27 theme sugar 902:906 arg1 UDP-Gal 914:920 its sugar donor UDP-Gal 898:920 its sugar donor UDP-Gal 898:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 5 28 theme front-side 1049:1058 arg1 reaction 1089:1096 a front-side substrate-assisted SN i-type reaction 1047:1096 a front-side substrate-assisted SN i-type reaction for α3GalT 1047:1107 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 4 29 dep Bos 778:780 arg1 taurus 782:787 Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal 778:920 taurus 782:787 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 29 dep Bos 778:780 arg1 contains 796:803 contains 796:803 contains 796:803 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 0 30 theme Native 57:62 arg1 Substrates 64:73 Native Substrates 57:73 Native Substrates 57:73 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 5 31 theme substrate-assisted 1060:1077 arg1 reaction 1089:1096 a front-side substrate-assisted SN i-type reaction 1047:1096 a front-side substrate-assisted SN i-type reaction for α3GalT 1047:1107 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 5 32 theme new 994:996 arg1 evidence 1011:1018 This new experimental evidence 989:1018 This new experimental evidence 989:1018 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 3 33 theme GTs 453:455 arg1 mechanism 430:438 the catalytic mechanism 416:438 the catalytic mechanism of retaining GTs 416:455 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 33 theme GTs 453:455 arg1 topic 462:466 a topic 460:466 a topic 460:466 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 34 theme cation 972:977 arg1 cofactor 979:986 the divalent cation cofactor 959:986 the divalent cation cofactor 959:986 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 5 35 theme common 1135:1140 arg1 mechanism 1152:1160 a conserved common catalytic mechanism 1123:1160 a conserved common catalytic mechanism among retaining GTs 1123:1180 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 4 36 theme UDP-Gal 914:920 arg1 presence 886:893 the presence 882:893 the presence of its sugar donor UDP-Gal 882:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 37 theme double-displacement 614:632 arg1 mechanism 634:642 a double-displacement mechanism 612:642 a double-displacement mechanism 612:642 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 5 38 theme experimental 998:1009 arg1 evidence 1011:1018 This new experimental evidence 989:1018 This new experimental evidence 989:1018 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 1 39 from synthesis 221:229 arg1 organisms 265:273 all living organisms 254:273 all living organisms 254:273 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 4 40 theme substrate 936:944 arg1 lactose 946:952 the acceptor substrate lactose 923:952 the acceptor substrate lactose 923:952 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 41 contain containing 521:530 arg2 residue 556:562 a putative nucleophilic residue 532:562 a putative nucleophilic residue in the active site 532:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 41 contain containing 521:530 arg1 enzymes 513:519 those enzymes 507:519 those enzymes containing a putative nucleophilic residue in the active site 507:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 42 theme mechanism 634:642 arg1 occurrence 598:607 the occurrence 594:607 the occurrence of a double-displacement mechanism 594:642 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 43 theme controversy 477:487 arg1 mechanism 430:438 the catalytic mechanism 416:438 the catalytic mechanism of retaining GTs 416:455 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 3 43 theme controversy 477:487 arg1 topic 462:466 a topic 460:466 a topic 460:466 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 44 theme active 831:836 arg1 site 838:841 the active site 827:841 the active site 827:841 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 0 45 with Snapshots 11:19 arg1 Substrates 64:73 Native Substrates 57:73 Native Substrates 57:73 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 5 46 theme SN 1079:1080 arg1 reaction 1089:1096 a front-side substrate-assisted SN i-type reaction 1047:1096 a front-side substrate-assisted SN i-type reaction for α3GalT 1047:1107 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 28960760 3 47 from residue 556:562 arg1 site 578:581 the active site 567:581 the active site 567:581 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 48 theme α-1,3-galactosyltransferase 736:762 arg1 complexes 689:697 native ternary complexes 674:697 native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal 674:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 48 theme α-1,3-galactosyltransferase 736:762 arg1 lactose 946:952 the acceptor substrate lactose 923:952 the acceptor substrate lactose 923:952 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 48 theme α-1,3-galactosyltransferase 736:762 arg1 cofactor 979:986 the divalent cation cofactor 959:986 the divalent cation cofactor 959:986 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 1 49 theme glycosidic 234:243 arg1 bonds 245:249 glycosidic bonds 234:249 glycosidic bonds 234:249 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 4 50 from nucleophile 812:822 arg1 site 838:841 the active site 827:841 the active site 827:841 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 51 theme great 471:475 arg1 controversy 477:487 great controversy 471:487 great controversy 471:487 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 1 52 theme bonds 245:249 arg1 synthesis 221:229 the synthesis 217:229 the synthesis of glycosidic bonds in all living organisms 217:273 Glycosyltransferases (GTs) are a key family of enzymes that catalyze the synthesis of glycosidic bonds in all living organisms. 28960760 0 53 dep Snapshots 11:19 arg1 Insight 76:82 Insight 76:82 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 0:146 Structural Snapshots of α-1,3-Galactosyltransferase with Native Substrates: Insight into the Catalytic Mechanism of Retaining Glycosyltransferases. 28960760 4 54 theme ternary 681:687 arg1 complexes 689:697 native ternary complexes 674:697 native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal 674:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 3 55 theme retaining 443:451 arg1 GTs 453:455 retaining GTs 443:455 retaining GTs 443:455 To date, the catalytic mechanism of retaining GTs is a topic of great controversy, particularly for those enzymes containing a putative nucleophilic residue in the active site, for which the occurrence of a double-displacement mechanism has been suggested. 28960760 4 56 contain contains 796:803 arg2 which 790:794 which 790:794 which 790:794 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 56 contain contains 796:803 arg2 nucleophile 812:822 such a nucleophile 805:822 such a nucleophile in the active site 805:841 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 56 contain contains 796:803 arg1 presence 886:893 the presence 882:893 the presence of its sugar donor UDP-Gal 882:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 56 contain contains 796:803 arg1 mode 860:863 a productive mode 847:863 a productive mode for catalysis 847:877 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 56 contain contains 796:803 arg1 which 790:794 which 790:794 which 790:794 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 4 57 theme native 674:679 arg1 complexes 689:697 native ternary complexes 674:697 native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal 674:920 We report native ternary complexes of the retaining glycosyltransferase α-1,3-galactosyltransferase (α3GalT) from Bos taurus, which contains such a nucleophile in the active site, in a productive mode for catalysis in the presence of its sugar donor UDP-Gal, the acceptor substrate lactose, and the divalent cation cofactor. 28960760 5 58 theme catalytic 1142:1150 arg1 mechanism 1152:1160 a conserved common catalytic mechanism 1123:1160 a conserved common catalytic mechanism among retaining GTs 1123:1180 This new experimental evidence supports the occurrence of a front-side substrate-assisted SN i-type reaction for α3GalT, and suggests a conserved common catalytic mechanism among retaining GTs. 23581857 0 0 theme arylsulfatase 68:80 arg1 ARSA 102:105 ARSA 102:105 ARSA 102:105 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 0 theme arylsulfatase 68:80 arg1 A 99:99 human arylsulfatase A. Arylsulfatase A 62:99 human arylsulfatase A. Arylsulfatase A (ARSA) 62:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 3 1 dep ARSAglyc 1015:1022 arg1 pH5 1024:1026 pH5 1024:1026 pH5 1024:1026 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 2 2 theme different 571:579 arg1 pH 581:582 (1) different pH's 567:584 (1) different pH's 567:584 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 7 3 theme ARSA 1713:1716 arg1 pseudodeficiency 1718:1733 ARSA pseudodeficiency 1713:1733 ARSA pseudodeficiency 1713:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 2 4 theme high 777:780 arg1 oligosaccharides 795:810 three high mannose-type oligosaccharides 771:810 three high mannose-type oligosaccharides 771:810 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 0 5 theme human 62:66 arg1 ARSA 102:105 ARSA 102:105 ARSA 102:105 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 5 theme human 62:66 arg1 A 99:99 human arylsulfatase A. Arylsulfatase A 62:99 human arylsulfatase A. Arylsulfatase A (ARSA) 62:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 7 6 theme molecular 1691:1699 arg1 etiology 1701:1708 molecular etiology 1691:1708 molecular etiology of ARSA pseudodeficiency 1691:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 2 7 theme natural 602:608 arg1 environment 621:631 its natural lysossomal environment 598:631 its natural lysossomal environment 598:631 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 7 8 theme defect 1798:1803 arg1 absence 1771:1777 the absence 1767:1777 the absence of polyadenylation defect 1767:1803 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 7 9 theme obtained 1549:1556 arg1 data 1558:1561 the obtained data 1545:1561 the obtained data 1545:1561 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 3 10 dep ARSAN350S 1067:1075 arg1 pH5 1077:1079 pH5 1077:1079 pH5 1077:1079 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 0 11 theme Arylsulfatase 85:97 arg1 ARSA 102:105 ARSA 102:105 ARSA 102:105 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 11 theme Arylsulfatase 85:97 arg1 A 99:99 human arylsulfatase A. Arylsulfatase A 62:99 human arylsulfatase A. Arylsulfatase A (ARSA) 62:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 12 from Effects 0:6 arg1 dynamics 50:57 the dynamics 46:57 the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) 46:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 13 theme cerebroside 166:176 arg1 sulfate 178:184 cerebroside sulfate 166:184 cerebroside sulfate 166:184 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 7 14 theme biological 1659:1668 arg1 activity 1670:1677 biological activity 1659:1677 biological activity of ARSA 1659:1685 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 0 15 theme A. 82:83 arg1 ARSA 102:105 ARSA 102:105 ARSA 102:105 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 15 theme A. 82:83 arg1 A 99:99 human arylsulfatase A. Arylsulfatase A 62:99 human arylsulfatase A. Arylsulfatase A (ARSA) 62:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 5 16 theme triple 1366:1371 arg1 glycosylation 1373:1385 triple glycosylation 1366:1385 triple glycosylation 1366:1385 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 7 17 theme ARSA 1682:1685 arg1 activity 1670:1677 biological activity 1659:1677 biological activity of ARSA 1659:1685 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 7 17 theme ARSA 1682:1685 arg1 etiology 1701:1708 molecular etiology 1691:1708 molecular etiology of ARSA pseudodeficiency 1691:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 2 18 theme glycan 698:703 arg1 state 753:757 its normal glycosylation state 728:757 its normal glycosylation state 728:757 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 2 18 theme glycan 698:703 arg1 occupancies 705:715 glycan occupancies 698:715 glycan occupancies 698:715 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 1 19 theme minor 253:257 arg1 condition 259:267 a minor condition 251:267 a minor condition called ARSA pseudodeficiency 251:296 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 5 20 theme nonglycosylated 1301:1315 arg1 counterpart 1317:1327 its nonglycosylated counterpart 1297:1327 its nonglycosylated counterpart 1297:1327 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 21 theme conformational 1255:1268 arg1 stability 1270:1278 a higher secondary conformational stability 1236:1278 a higher secondary conformational stability 1236:1278 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 3 22 theme different 875:883 arg1 conditions 885:894 different conditions 875:894 different conditions 875:894 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 7 23 theme etiology 1701:1708 arg1 understanding 1642:1654 the understanding 1638:1654 the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency 1638:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 0 24 theme A 99:99 arg1 dynamics 50:57 the dynamics 46:57 the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) 46:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 25 theme sulfate 178:184 arg1 hydrolysis 152:161 the hydrolysis 148:161 the hydrolysis of cerebroside sulfate 148:184 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 5 26 theme higher 1238:1243 arg1 stability 1270:1278 a higher secondary conformational stability 1236:1278 a higher secondary conformational stability 1236:1278 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 3 27 gly glycosylated 991:1002 arg2 pH ∼ 5 1007:1012 pH ∼ 5 1007:1012 pH ∼ 5 1007:1012 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 27 gly glycosylated 991:1002 arg2 pH ∼ 5 1059:1064 pH ∼ 5 1059:1064 pH ∼ 5 (ARSAN350S,pH5) 1059:1080 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 27 gly glycosylated 991:1002 arg2 ARSAglyc 1015:1022 ARSAglyc 1015:1022 ARSAglyc 1015:1022 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 27 gly glycosylated 991:1002 arg2 mutant 1049:1054 (4) ARSA-N350S mutant 1034:1054 (4) ARSA-N350S mutant 1034:1054 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 27 gly glycosylated 991:1002 arg2 ARSAN350S 1067:1075 ARSAN350S 1067:1075 ARSAN350S 1067:1075 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 4 28 theme enzyme 1167:1172 arg1 flexibility 1148:1158 the flexibility 1144:1158 the flexibility of the enzyme 1144:1172 Lowering pH and increasing glycosylation was found to reduce the flexibility of the enzyme. 23581857 1 29 theme polyadenylation 438:452 arg1 signal 454:459 the polyadenylation signal 434:459 the polyadenylation signal 434:459 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 2 30 dep pH 581:582 arg1 response 555:562 response 555:562 response 555:562 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 2 30 dep pH 581:582 arg1 1 568:568 1 568:568 1 568:568 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 5 31 theme experimental 1341:1352 arg1 findings 1354:1361 experimental findings 1341:1361 experimental findings on triple glycosylation 1341:1385 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 31 theme experimental 1341:1352 arg1 state 1404:1408 the essential state 1390:1408 the essential state of ARSA 1390:1416 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 1 32 theme signal 454:459 arg1 loss 426:429 the loss 422:429 the loss of the polyadenylation signal 422:459 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 3 33 from triple 984:989 arg1 non-glycosylated 942:957 non-glycosylated 942:957 non-glycosylated 942:957 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 2 34 theme dynamics 531:538 arg1 simulations 540:550 molecular dynamics simulations 521:550 molecular dynamics simulations 521:550 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 2 35 theme lysossomal 610:619 arg1 environment 621:631 its natural lysossomal environment 598:631 its natural lysossomal environment 598:631 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 6 36 theme unfolding 1469:1477 arg1 related 1493:1499 related 1493:1499 related 1493:1499 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 6 36 theme unfolding 1469:1477 arg1 degree 1459:1464 a consistent degree 1446:1464 a consistent degree 1446:1464 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 0 37 theme glycosylation 11:23 arg1 sulfatase 123:131 a lysosomal sulfatase 111:131 a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate 111:184 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 37 theme glycosylation 11:23 arg1 Effects 0:6 Effects 0:6 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) 0:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 1 38 theme ARSA 276:279 arg1 pseudodeficiency 281:296 ARSA pseudodeficiency 276:296 ARSA pseudodeficiency 276:296 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 2 39 theme molecular 521:529 arg1 simulations 540:550 molecular dynamics simulations 521:550 molecular dynamics simulations 521:550 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 7 40 theme able 1616:1619 arg1 evidences 1606:1614 structural evidences 1595:1614 structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency 1595:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 3 41 gly nonglycosylated 901:915 arg2 ARSApH7 928:934 ARSApH7 928:934 ARSApH7 928:934 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 41 gly nonglycosylated 901:915 arg2 pH ∼ 7 920:925 pH ∼ 7 920:925 pH ∼ 7 (ARSApH7) 920:935 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 5 42 gly nonglycosylated 1301:1315 arg1 counterpart 1317:1327 its nonglycosylated counterpart 1297:1327 its nonglycosylated counterpart 1297:1327 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 0 43 theme conditions 32:41 arg1 sulfatase 123:131 a lysosomal sulfatase 111:131 a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate 111:184 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 43 theme conditions 32:41 arg1 Effects 0:6 Effects 0:6 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) 0:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 3 44 dep nonglycosylated 901:915 arg1 non-glycosylated 942:957 non-glycosylated 942:957 non-glycosylated 942:957 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 44 dep nonglycosylated 901:915 arg1 2 939:939 2 939:939 2 939:939 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 3 44 dep nonglycosylated 901:915 arg1 1 898:898 1 898:898 1 898:898 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 5 45 theme ARSA 1413:1416 arg1 findings 1354:1361 experimental findings 1341:1361 experimental findings on triple glycosylation 1341:1385 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 45 theme ARSA 1413:1416 arg1 state 1404:1408 the essential state 1390:1408 the essential state of ARSA 1390:1416 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 2 46 theme normal 732:737 arg1 state 753:757 its normal glycosylation state 728:757 its normal glycosylation state 728:757 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 5 47 attach presented 1226:1234 arg1 addition 1178:1185 addition 1178:1185 addition 1178:1185 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 47 attach presented 1226:1234 arg2 enzyme 1219:1224 the glycosylated enzyme 1202:1224 the glycosylated enzyme 1202:1224 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 47 attach presented 1226:1234 arg1 pH 1198:1199 acidic pH 1191:1199 acidic pH 1191:1199 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 0 48 theme pH 29:30 arg1 conditions 32:41 pH conditions 29:41 pH conditions 29:41 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 3 49 dep triple 984:989 arg1 3 981:981 3 981:981 3 981:981 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 1 50 theme glycosylated 384:395 arg1 Asn350 397:402 the glycosylated Asn350 380:402 the glycosylated Asn350 380:402 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 5 51 theme glycosylated 1206:1217 arg1 enzyme 1219:1224 the glycosylated enzyme 1202:1224 the glycosylated enzyme 1202:1224 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 52 theme secondary 1245:1253 arg1 stability 1270:1278 a higher secondary conformational stability 1236:1278 a higher secondary conformational stability 1236:1278 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 2 53 theme ARSA 497:500 arg1 dynamics 502:509 ARSA dynamics 497:509 ARSA dynamics 497:509 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 3 54 dep mutant 1049:1054 arg1 4 1035:1035 4 1035:1035 4 1035:1035 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 1 55 theme Asn350 397:402 arg1 substitution 364:375 the substitution 360:375 the substitution of the glycosylated Asn350 by a Ser 360:411 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 7 56 theme pseudodeficiency 1718:1733 arg1 activity 1670:1677 biological activity 1659:1677 biological activity of ARSA 1659:1685 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 7 56 theme pseudodeficiency 1718:1733 arg1 etiology 1701:1708 molecular etiology 1691:1708 molecular etiology of ARSA pseudodeficiency 1691:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 2 57 dep occupancies 705:715 arg1 2 695:695 2 695:695 2 695:695 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 1 58 theme healthy 308:314 arg1 individuals 316:326 healthy individuals 308:326 healthy individuals 308:326 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 0 59 theme lysosomal 113:121 arg1 sulfatase 123:131 a lysosomal sulfatase 111:131 a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate 111:184 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 0 59 theme lysosomal 113:121 arg1 Effects 0:6 Effects 0:6 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) 0:106 Effects of glycosylation and pH conditions in the dynamics of human arylsulfatase A. Arylsulfatase A (ARSA) is a lysosomal sulfatase that catalyzes the hydrolysis of cerebroside sulfate. 23581857 7 60 from ARSA-N350S 1753:1762 arg1 absence 1771:1777 the absence 1767:1777 the absence of polyadenylation defect 1767:1803 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 6 61 theme consistent 1448:1457 arg1 related 1493:1499 related 1493:1499 related 1493:1499 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 6 61 theme consistent 1448:1457 arg1 degree 1459:1464 a consistent degree 1446:1464 a consistent degree 1446:1464 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 7 62 theme polyadenylation 1782:1796 arg1 defect 1798:1803 polyadenylation defect 1782:1803 polyadenylation defect 1782:1803 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 5 63 from findings 1354:1361 arg1 glycosylation 1373:1385 triple glycosylation 1366:1385 triple glycosylation 1366:1385 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 6 64 theme reduced 1517:1523 arg1 stability 1525:1533 its in vitro reduced stability 1504:1533 its in vitro reduced stability 1504:1533 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 6 65 theme N350S 1423:1427 arg1 mutant 1429:1434 The N350S mutant 1419:1434 The N350S mutant 1419:1434 The N350S mutant exhibited a consistent degree of unfolding, which may be related to its in vitro reduced stability. 23581857 5 66 theme essential 1394:1402 arg1 findings 1354:1361 experimental findings 1341:1361 experimental findings on triple glycosylation 1341:1385 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 5 66 theme essential 1394:1402 arg1 state 1404:1408 the essential state 1390:1408 the essential state of ARSA 1390:1416 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 7 67 theme structural 1595:1604 arg1 evidences 1606:1614 structural evidences 1595:1614 structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency 1595:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 5 68 theme acidic 1191:1196 arg1 pH 1198:1199 acidic pH 1191:1199 acidic pH 1191:1199 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 2 69 theme glycosylation 739:751 arg1 state 753:757 its normal glycosylation state 728:757 its normal glycosylation state 728:757 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 7 70 theme activity 1670:1677 arg1 understanding 1642:1654 the understanding 1638:1654 the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency 1638:1733 Finally, the obtained data are discussed in the search for structural evidences able to contribute to the understanding of biological activity of ARSA and molecular etiology of ARSA pseudodeficiency, as determined by ARSA-N350S in the absence of polyadenylation defect. 23581857 3 71 gly non-glycosylated 942:957 arg2 triple 984:989 triple 984:989 triple 984:989 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 1 72 gly glycosylated 384:395 arg1 Asn350 397:402 the glycosylated Asn350 380:402 the glycosylated Asn350 380:402 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 1 73 theme Metachromatic 213:225 arg1 Leukodystrophy 227:240 Metachromatic Leukodystrophy 213:240 Metachromatic Leukodystrophy 213:240 Its deficiency results in Metachromatic Leukodystrophy, whereas a minor condition called ARSA pseudodeficiency occurs in healthy individuals, which has been associated with the substitution of the glycosylated Asn350 by a Ser and with the loss of the polyadenylation signal. 23581857 3 74 theme ARSA-N350S 1038:1047 arg1 mutant 1049:1054 (4) ARSA-N350S mutant 1034:1054 (4) ARSA-N350S mutant 1034:1054 Accordingly, four systems were studied considering ARSA under different conditions: (1) nonglycosylated at pH ∼ 7 (ARSApH7); (2) non-glycosylated at pH ∼ 5 (ARSApH5); (3) triple glycosylated at pH ∼ 5 (ARSAglyc,pH5); and (4) ARSA-N350S mutant at pH ∼ 5 (ARSAN350S,pH5). 23581857 2 75 theme cytoplasmatic 669:681 arg1 medium 683:688 cytoplasmatic medium 669:688 cytoplasmatic medium 669:688 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 23581857 5 76 gly glycosylated 1206:1217 arg1 enzyme 1219:1224 the glycosylated enzyme 1202:1224 the glycosylated enzyme 1202:1224 In addition, at acidic pH, the glycosylated enzyme presented a higher secondary conformational stability when compared to its nonglycosylated counterpart, supporting experimental findings on triple glycosylation as the essential state of ARSA. 23581857 2 77 theme mannose-type 782:793 arg1 oligosaccharides 795:810 three high mannose-type oligosaccharides 771:810 three high mannose-type oligosaccharides 771:810 In this work, we have investigated ARSA dynamics employing molecular dynamics simulations in response to (1) different pH's, as, beyond its natural lysossomal environment, it has been recently identified in cytoplasmatic medium and (2) glycan occupancies, including its normal glycosylation state, presenting three high mannose-type oligosaccharides. 24295106 1 0 theme reliable 321:328 arg1 methods 330:336 more sensitive and reliable methods 302:336 more sensitive and reliable methods for early detection of the disease 302:371 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 0 1 theme cell 209:212 arg1 lines 214:218 the more metastatic/aggressive cell lines 178:218 the more metastatic/aggressive cell lines 178:218 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 2 2 theme cell 794:797 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 8 3 theme cell 1936:1939 arg1 lines 1941:1945 the other two cell lines 1922:1945 the other two cell lines 1922:1945 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 4 4 theme mannose 1069:1075 arg1 % 1106:1106 70-90% 1101:1106 70-90% 1101:1106 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 4 theme mannose 1069:1075 arg1 structures 1089:1098 high mannose type glycan structures 1064:1098 high mannose type glycan structures (70-90%) 1064:1107 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 5 5 located observed 1261:1268 arg2 N-acetylglucosamine 1236:1254 bisecting N-acetylglucosamine 1226:1254 bisecting N-acetylglucosamine 1226:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 5 located observed 1261:1268 arg1 level 1280:1284 a high level 1273:1284 a high level in the metastatic LIM1215 cells 1273:1316 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 5 located observed 1261:1268 arg2 determinants 1205:1216 unique glycan determinants 1191:1216 unique glycan determinants such as bisecting N-acetylglucosamine 1191:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 2 6 theme primary 782:788 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 4 7 theme key 976:978 arg1 landscape 1017:1025 the similar N-glycan landscape 996:1025 the similar N-glycan landscape in these cells 996:1040 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 7 theme key 976:978 arg1 observation 980:990 A key observation 974:990 A key observation 974:990 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 8 8 theme accumulation 1829:1840 arg1 degree 1784:1789 the higher degree 1773:1789 the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans 1773:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 8 9 theme incomplete 1794:1803 arg1 processing 1814:1823 incomplete N-glycan processing 1794:1823 incomplete N-glycan processing 1794:1823 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 10 theme proteins 565:572 arg1 profiles 544:551 N-glycan profiles 535:551 N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively 535:817 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 9 11 theme membrane 2000:2007 arg1 N-glycome 2009:2017 the membrane N-glycome 1996:2017 the membrane N-glycome in three CRC cell lines 1996:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 8 12 theme processing 1814:1823 arg1 degree 1784:1789 the higher degree 1773:1789 the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans 1773:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 1 13 theme colorectal 233:242 arg1 CRC 252:254 CRC 252:254 CRC 252:254 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 1 13 theme colorectal 233:242 arg1 cancer 244:249 colorectal cancer 233:249 colorectal cancer (CRC) diagnosis 233:265 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 2 14 theme cell 614:617 arg1 LIM1215 626:632 LIM1215 626:632 LIM1215 626:632 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 14 theme cell 614:617 arg1 lines 619:623 three phenotypically different CRC cell lines 579:623 three phenotypically different CRC cell lines 579:623 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 14 theme cell 614:617 arg1 LIM1899 635:641 LIM1899 635:641 LIM1899 635:641 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 14 theme cell 614:617 arg1 LIM2405 648:654 LIM2405 648:654 LIM2405 648:654 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 9 15 from analysis 1984:1991 arg1 lines 2037:2041 three CRC cell lines 2022:2041 three CRC cell lines 2022:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 2 16 theme different 600:608 arg1 LIM1215 626:632 LIM1215 626:632 LIM1215 626:632 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 16 theme different 600:608 arg1 lines 619:623 three phenotypically different CRC cell lines 579:623 three phenotypically different CRC cell lines 579:623 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 16 theme different 600:608 arg1 LIM1899 635:641 LIM1899 635:641 LIM1899 635:641 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 16 theme different 600:608 arg1 LIM2405 648:654 LIM2405 648:654 LIM2405 648:654 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 0 17 theme membrane 157:164 arg1 proteins 166:173 membrane proteins 157:173 membrane proteins of the more metastatic/aggressive cell lines 157:218 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 2 18 with glycoproteins 464:476 arg1 functions 504:512 important biological functions 483:512 important biological functions 483:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 3 19 theme underivatized 899:911 arg1 abundances 863:872 their relative abundances 848:872 their relative abundances 848:872 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 19 theme underivatized 899:911 arg1 structures 833:842 The N-glycan structures 820:842 The N-glycan structures 820:842 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 19 theme underivatized 899:911 arg1 forms 921:925 their underivatized reduced forms 893:925 their underivatized reduced forms 893:925 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 7 20 theme Mgat3 1666:1670 arg1 binding 1676:1682 binding 1676:1682 binding 1676:1682 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 7 20 theme Mgat3 1666:1670 arg1 upregulation 1650:1661 the highest upregulation 1638:1661 the highest upregulation of Mgat3 1638:1670 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 6 21 attach present 1529:1535 arg2 α-2,3-sialylation 1462:1478 α-2,3-sialylation 1462:1478 α-2,3-sialylation 1462:1478 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 6 21 attach present 1529:1535 arg1 LIM2405 1545:1551 LIM2405 1545:1551 LIM2405 1545:1551 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 9 22 theme CRC 2028:2030 arg1 lines 2037:2041 three CRC cell lines 2022:2041 three CRC cell lines 2022:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 10 23 theme molecular 2229:2237 arg1 phenotypes 2193:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 10 23 theme molecular 2229:2237 arg1 feature 2239:2245 a molecular feature 2227:2245 a molecular feature to differentiate CRC disease stages 2227:2281 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 8 24 theme high 1845:1848 arg1 structures 1863:1872 high mannose type structures 1845:1872 high mannose type structures as well as bisecting N-glycans 1845:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 25 dep differentiated 719:732 arg1 primary 734:740 primary 734:740 primary 734:740 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 3 26 theme N-glycan 824:831 arg1 abundances 863:872 their relative abundances 848:872 their relative abundances 848:872 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 26 theme N-glycan 824:831 arg1 structures 833:842 The N-glycan structures 820:842 The N-glycan structures 820:842 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 26 theme N-glycan 824:831 arg1 forms 921:925 their underivatized reduced forms 893:925 their underivatized reduced forms 893:925 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 8 27 theme type 1858:1861 arg1 structures 1863:1872 high mannose type structures 1845:1872 high mannose type structures as well as bisecting N-glycans 1845:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 28 theme differentiated 754:767 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 5 29 theme metastatic 1293:1302 arg1 cells 1312:1316 the metastatic LIM1215 cells 1289:1316 the metastatic LIM1215 cells 1289:1316 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 0 30 theme bisecting 78:86 arg1 GlcNAc 88:93 unique bisecting GlcNAc 71:93 unique bisecting GlcNAc 71:93 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 8 31 theme other 1926:1930 arg1 lines 1941:1945 the other two cell lines 1922:1945 the other two cell lines 1922:1945 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 32 dep differentiated 681:694 arg1 aggressive 770:779 aggressive 770:779 aggressive 770:779 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 32 dep differentiated 681:694 arg1 metastatic 696:705 metastatic 696:705 metastatic 696:705 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 4 33 theme similar 1000:1006 arg1 landscape 1017:1025 the similar N-glycan landscape 996:1025 the similar N-glycan landscape in these cells 996:1040 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 33 theme similar 1000:1006 arg1 observation 980:990 A key observation 974:990 A key observation 974:990 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 3 34 theme relative 854:861 arg1 abundances 863:872 their relative abundances 848:872 their relative abundances 848:872 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 34 theme relative 854:861 arg1 structures 833:842 The N-glycan structures 820:842 The N-glycan structures 820:842 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 34 theme relative 854:861 arg1 forms 921:925 their underivatized reduced forms 893:925 their underivatized reduced forms 893:925 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 9 35 theme N-glycosylation 2058:2072 arg1 differences 2074:2084 N-glycosylation differences 2058:2084 N-glycosylation differences that correlate with the histological and pathological features of the cell lines 2058:2165 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 2 36 theme N-glycan 535:542 arg1 profiles 544:551 N-glycan profiles 535:551 N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively 535:817 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 8 37 from Downregulation 1707:1720 arg1 LIM1215 1745:1751 LIM1215 1745:1751 LIM1215 1745:1751 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 0 38 theme sialic 112:117 arg1 determinants 124:135 α-2,3-linked sialic acid determinants 99:135 α-2,3-linked sialic acid determinants 99:135 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 7 39 from PHA-E 1689:1693 arg1 LIM1215 1698:1704 LIM1215 1698:1704 LIM1215 1698:1704 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 8 40 theme N-glycans 1895:1903 arg1 accumulation 1829:1840 accumulation 1829:1840 accumulation of high mannose type structures as well as bisecting N-glycans 1829:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 8 40 theme N-glycans 1895:1903 arg1 processing 1814:1823 incomplete N-glycan processing 1794:1823 incomplete N-glycan processing 1794:1823 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 3 41 theme graphitized 941:951 arg1 LC-ESI-MS/MS 960:971 porous graphitized carbon LC-ESI-MS/MS 934:971 porous graphitized carbon LC-ESI-MS/MS 934:971 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 4 42 theme type 1077:1080 arg1 % 1106:1106 70-90% 1101:1106 70-90% 1101:1106 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 42 theme type 1077:1080 arg1 structures 1089:1098 high mannose type glycan structures 1064:1098 high mannose type glycan structures (70-90%) 1064:1107 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 0 43 theme Comparative 0:10 arg1 profiling 21:29 Comparative N-glycan profiling 0:29 Comparative N-glycan profiling of colorectal cancer cell lines 0:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 9 44 from lines 2037:2041 arg1 analysis 1984:1991 a comprehensive analysis 1968:1991 a comprehensive analysis of the membrane N-glycome in three CRC cell lines 1968:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 4 45 theme structures 1089:1098 arg1 dominance 1051:1059 the dominance 1047:1059 the dominance of high mannose type glycan structures (70-90%) in all three cell lines 1047:1131 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 2 46 from lines 619:623 arg1 profiles 544:551 N-glycan profiles 535:551 N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively 535:817 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 47 theme disease 426:432 arg1 biomarkers 434:443 many known disease biomarkers 415:443 many known disease biomarkers 415:443 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 47 theme disease 426:432 arg1 glycoproteins 464:476 membrane-bound glycoproteins 449:476 membrane-bound glycoproteins with important biological functions 449:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 4 48 theme incomplete 1148:1157 arg1 processing 1166:1175 an incomplete glycan processing 1145:1175 an incomplete glycan processing 1145:1175 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 5 49 theme differentiated 1357:1370 arg1 LIM1899 1372:1378 the moderately differentiated LIM1899 1342:1378 the moderately differentiated LIM1899 1342:1378 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 2 50 theme many 415:418 arg1 biomarkers 434:443 many known disease biomarkers 415:443 many known disease biomarkers 415:443 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 50 theme many 415:418 arg1 glycoproteins 464:476 membrane-bound glycoproteins 449:476 membrane-bound glycoproteins with important biological functions 449:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 0 51 theme colorectal 34:43 arg1 lines 57:61 colorectal cancer cell lines 34:61 colorectal cancer cell lines 34:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 1 52 theme sensitive 307:315 arg1 methods 330:336 more sensitive and reliable methods 302:336 more sensitive and reliable methods for early detection of the disease 302:371 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 7 53 theme immunofluorescence 1573:1590 arg1 data 1592:1595 lectin immunofluorescence data 1566:1595 lectin immunofluorescence data 1566:1595 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 6 54 from present 1529:1535 arg1 LIM2405 1545:1551 LIM2405 1545:1551 LIM2405 1545:1551 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 4 55 theme high 1064:1067 arg1 % 1106:1106 70-90% 1101:1106 70-90% 1101:1106 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 55 theme high 1064:1067 arg1 structures 1089:1098 high mannose type glycan structures 1064:1098 high mannose type glycan structures (70-90%) 1064:1107 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 5 56 theme differentiated 1420:1433 arg1 cells 1443:1447 the poorly differentiated LIM2405 cells 1409:1447 the poorly differentiated LIM2405 cells 1409:1447 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 0 57 theme cell 52:55 arg1 lines 57:61 colorectal cancer cell lines 34:61 colorectal cancer cell lines 34:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 5 58 theme unique 1191:1196 arg1 N-acetylglucosamine 1236:1254 bisecting N-acetylglucosamine 1226:1254 bisecting N-acetylglucosamine 1226:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 58 theme unique 1191:1196 arg1 determinants 1205:1216 unique glycan determinants 1191:1216 unique glycan determinants such as bisecting N-acetylglucosamine 1191:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 9 59 theme lines 2161:2165 arg1 features 2140:2147 the histological and pathological features 2106:2147 the histological and pathological features of the cell lines 2106:2165 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 1 60 theme cancer 244:249 arg1 diagnosis 257:265 colorectal cancer (CRC) diagnosis 233:265 colorectal cancer (CRC) diagnosis 233:265 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 6 61 from LIM1215 1505:1511 arg1 absent 1495:1500 absent 1495:1500 absent 1495:1500 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 4 62 with landscape 1017:1025 arg1 dominance 1051:1059 the dominance 1047:1059 the dominance of high mannose type glycan structures (70-90%) in all three cell lines 1047:1131 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 63 theme cell 1122:1125 arg1 lines 1127:1131 all three cell lines 1112:1131 all three cell lines 1112:1131 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 10 64 theme glycosylation 2179:2191 arg1 phenotypes 2193:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 10 64 theme glycosylation 2179:2191 arg1 feature 2239:2245 a molecular feature 2227:2245 a molecular feature to differentiate CRC disease stages 2227:2281 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 2 65 theme important 483:491 arg1 functions 504:512 important biological functions 483:512 important biological functions 483:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 6 66 from LIM1899 1517:1523 arg1 absent 1495:1500 absent 1495:1500 absent 1495:1500 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 0 67 link α-2,3-linked 99:110 arg1 determinants 124:135 α-2,3-linked sialic acid determinants 99:135 α-2,3-linked sialic acid determinants 99:135 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 0 68 theme metastatic/aggressive 187:207 arg1 lines 214:218 the more metastatic/aggressive cell lines 178:218 the more metastatic/aggressive cell lines 178:218 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 4 69 from landscape 1017:1025 arg1 cells 1036:1040 these cells 1030:1040 these cells 1030:1040 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 5 70 located detected 1397:1404 arg2 none 1387:1390 none 1387:1390 none 1387:1390 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 70 located detected 1397:1404 arg1 cells 1443:1447 the poorly differentiated LIM2405 cells 1409:1447 the poorly differentiated LIM2405 cells 1409:1447 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 0 71 theme lines 214:218 arg1 proteins 166:173 membrane proteins 157:173 membrane proteins of the more metastatic/aggressive cell lines 157:218 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 10 72 theme CRC 2264:2266 arg1 stages 2276:2281 CRC disease stages 2264:2281 CRC disease stages 2264:2281 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 4 73 theme glycan 1159:1164 arg1 processing 1166:1175 an incomplete glycan processing 1145:1175 an incomplete glycan processing 1145:1175 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 9 74 theme comprehensive 1970:1982 arg1 analysis 1984:1991 a comprehensive analysis 1968:1991 a comprehensive analysis of the membrane N-glycome in three CRC cell lines 1968:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 2 75 theme CRC 790:792 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 4 76 from dominance 1051:1059 arg1 lines 1127:1131 all three cell lines 1112:1131 all three cell lines 1112:1131 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 8 77 theme higher 1777:1782 arg1 degree 1784:1789 the higher degree 1773:1789 the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans 1773:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 78 theme membrane 556:563 arg1 proteins 565:572 membrane proteins 556:572 membrane proteins 556:572 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 9 79 theme N-glycome 2009:2017 arg1 analysis 1984:1991 a comprehensive analysis 1968:1991 a comprehensive analysis of the membrane N-glycome in three CRC cell lines 1968:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 1 80 theme methods 330:336 arg1 development 287:297 development 287:297 development of more sensitive and reliable methods for early detection of the disease 287:371 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 5 81 from level 1280:1284 arg1 cells 1312:1316 the metastatic LIM1215 cells 1289:1316 the metastatic LIM1215 cells 1289:1316 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 6 82 from absent 1495:1500 arg1 LIM1899 1517:1523 LIM1899 1517:1523 LIM1899 1517:1523 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 6 82 from absent 1495:1500 arg1 LIM1215 1505:1511 LIM1215 1505:1511 LIM1215 1505:1511 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 8 83 theme N-glycan 1805:1812 arg1 processing 1814:1823 incomplete N-glycan processing 1794:1823 incomplete N-glycan processing 1794:1823 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 2 84 theme CRC 610:612 arg1 LIM1215 626:632 LIM1215 626:632 LIM1215 626:632 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 84 theme CRC 610:612 arg1 lines 619:623 three phenotypically different CRC cell lines 579:623 three phenotypically different CRC cell lines 579:623 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 84 theme CRC 610:612 arg1 LIM1899 635:641 LIM1899 635:641 LIM1899 635:641 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 84 theme CRC 610:612 arg1 LIM2405 648:654 LIM2405 648:654 LIM2405 648:654 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 1 85 from Advances 221:228 arg1 diagnosis 257:265 colorectal cancer (CRC) diagnosis 233:265 colorectal cancer (CRC) diagnosis 233:265 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 3 86 theme reduced 913:919 arg1 abundances 863:872 their relative abundances 848:872 their relative abundances 848:872 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 86 theme reduced 913:919 arg1 structures 833:842 The N-glycan structures 820:842 The N-glycan structures 820:842 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 3 86 theme reduced 913:919 arg1 forms 921:925 their underivatized reduced forms 893:925 their underivatized reduced forms 893:925 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 7 87 theme highest 1642:1648 arg1 upregulation 1650:1661 the highest upregulation 1638:1661 the highest upregulation of Mgat3 1638:1670 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 9 88 theme cell 2032:2035 arg1 lines 2037:2041 three CRC cell lines 2022:2041 three CRC cell lines 2022:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 0 89 theme unique 71:76 arg1 GlcNAc 88:93 unique bisecting GlcNAc 71:93 unique bisecting GlcNAc 71:93 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 10 90 theme disease 2268:2274 arg1 stages 2276:2281 CRC disease stages 2264:2281 CRC disease stages 2264:2281 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 1 91 theme early 342:346 arg1 detection 348:356 early detection 342:356 early detection of the disease 342:371 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 8 92 theme mannose 1850:1856 arg1 structures 1863:1872 high mannose type structures 1845:1872 high mannose type structures as well as bisecting N-glycans 1845:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 3 93 theme porous 934:939 arg1 LC-ESI-MS/MS 960:971 porous graphitized carbon LC-ESI-MS/MS 934:971 porous graphitized carbon LC-ESI-MS/MS 934:971 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 2 94 theme differentiated 681:694 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 5 95 theme LIM1215 1304:1310 arg1 cells 1312:1316 the metastatic LIM1215 cells 1289:1316 the metastatic LIM1215 cells 1289:1316 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 0 96 theme α-2,3-linked 99:110 arg1 determinants 124:135 α-2,3-linked sialic acid determinants 99:135 α-2,3-linked sialic acid determinants 99:135 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 2 97 gly glycoproteins 464:476 arg1 biomarkers 434:443 many known disease biomarkers 415:443 many known disease biomarkers 415:443 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 97 gly glycoproteins 464:476 arg1 glycoproteins 464:476 membrane-bound glycoproteins 449:476 membrane-bound glycoproteins with important biological functions 449:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 8 98 theme structures 1863:1872 arg1 accumulation 1829:1840 accumulation 1829:1840 accumulation of high mannose type structures as well as bisecting N-glycans 1829:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 8 98 theme structures 1863:1872 arg1 processing 1814:1823 incomplete N-glycan processing 1794:1823 incomplete N-glycan processing 1794:1823 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 10 99 theme unique 2172:2177 arg1 phenotypes 2193:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes 2168:2202 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 10 99 theme unique 2172:2177 arg1 feature 2239:2245 a molecular feature 2227:2245 a molecular feature to differentiate CRC disease stages 2227:2281 The unique glycosylation phenotypes may therefore serve as a molecular feature to differentiate CRC disease stages. 24295106 0 100 theme acid 119:122 arg1 determinants 124:135 α-2,3-linked sialic acid determinants 99:135 α-2,3-linked sialic acid determinants 99:135 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 1 101 theme disease 365:371 arg1 detection 348:356 early detection 342:356 early detection of the disease 342:371 Advances in colorectal cancer (CRC) diagnosis will be enhanced by development of more sensitive and reliable methods for early detection of the disease when treatment is more effective. 24295106 4 102 theme N-glycan 1008:1015 arg1 landscape 1017:1025 the similar N-glycan landscape 996:1025 the similar N-glycan landscape in these cells 996:1040 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 102 theme N-glycan 1008:1015 arg1 observation 980:990 A key observation 974:990 A key observation 974:990 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 2 103 theme differentiated 719:732 arg1 lines 799:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines 670:803 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 0 104 theme N-glycan 12:19 arg1 profiling 21:29 Comparative N-glycan profiling 0:29 Comparative N-glycan profiling of colorectal cancer cell lines 0:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 8 105 theme bisecting 1885:1893 arg1 N-glycans 1895:1903 bisecting N-glycans 1885:1903 high mannose type structures as well as bisecting N-glycans 1845:1903 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 3 106 theme carbon 953:958 arg1 LC-ESI-MS/MS 960:971 porous graphitized carbon LC-ESI-MS/MS 934:971 porous graphitized carbon LC-ESI-MS/MS 934:971 The N-glycan structures and their relative abundances were determined as their underivatized reduced forms, using porous graphitized carbon LC-ESI-MS/MS. 24295106 2 107 theme membrane-bound 449:462 arg1 biomarkers 434:443 many known disease biomarkers 415:443 many known disease biomarkers 415:443 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 107 theme membrane-bound 449:462 arg1 glycoproteins 464:476 membrane-bound glycoproteins 449:476 membrane-bound glycoproteins with important biological functions 449:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 9 108 theme histological 2110:2121 arg1 features 2140:2147 the histological and pathological features 2106:2147 the histological and pathological features of the cell lines 2106:2165 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 4 109 theme glycan 1082:1087 arg1 % 1106:1106 70-90% 1101:1106 70-90% 1101:1106 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 4 109 theme glycan 1082:1087 arg1 structures 1089:1098 high mannose type glycan structures 1064:1098 high mannose type glycan structures (70-90%) 1064:1107 A key observation was the similar N-glycan landscape in these cells with the dominance of high mannose type glycan structures (70-90%) in all three cell lines, suggesting an incomplete glycan processing. 24295106 9 110 theme pathological 2127:2138 arg1 features 2140:2147 the histological and pathological features 2106:2147 the histological and pathological features of the cell lines 2106:2165 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 2 111 theme known 420:424 arg1 biomarkers 434:443 many known disease biomarkers 415:443 many known disease biomarkers 415:443 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 2 111 theme known 420:424 arg1 glycoproteins 464:476 membrane-bound glycoproteins 449:476 membrane-bound glycoproteins with important biological functions 449:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 0 112 theme cancer 45:50 arg1 lines 57:61 colorectal cancer cell lines 34:61 colorectal cancer cell lines 34:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 5 113 theme bisecting 1226:1234 arg1 N-acetylglucosamine 1236:1254 bisecting N-acetylglucosamine 1226:1254 bisecting N-acetylglucosamine 1226:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 9 114 from N-glycome 2009:2017 arg1 lines 2037:2041 three CRC cell lines 2022:2041 three CRC cell lines 2022:2041 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 2 115 theme biological 493:502 arg1 functions 504:512 important biological functions 483:512 important biological functions 483:512 Because many known disease biomarkers are membrane-bound glycoproteins with important biological functions, we chose to compare N-glycan profiles of membrane proteins from three phenotypically different CRC cell lines, LIM1215, LIM1899, and LIM2405, representing moderately differentiated metastatic, moderately differentiated primary, and poorly differentiated (aggressive) primary CRC cell lines, respectively. 24295106 0 116 theme lines 57:61 arg1 profiling 21:29 Comparative N-glycan profiling 0:29 Comparative N-glycan profiling of colorectal cancer cell lines 0:61 Comparative N-glycan profiling of colorectal cancer cell lines reveals unique bisecting GlcNAc and α-2,3-linked sialic acid determinants are associated with membrane proteins of the more metastatic/aggressive cell lines. 24295106 6 117 from LIM2405 1545:1551 arg1 present 1529:1535 present 1529:1535 present 1529:1535 Conversely, α-2,3-sialylation was completely absent in LIM1215 and LIM1899 and present only in LIM2405. 24295106 9 118 theme cell 2156:2159 arg1 lines 2161:2165 the cell lines 2152:2165 the cell lines 2152:2165 This study provides a comprehensive analysis of the membrane N-glycome in three CRC cell lines and identifies N-glycosylation differences that correlate with the histological and pathological features of the cell lines. 24295106 5 119 theme LIM2405 1435:1441 arg1 cells 1443:1447 the poorly differentiated LIM2405 cells 1409:1447 the poorly differentiated LIM2405 cells 1409:1447 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 120 theme glycan 1198:1203 arg1 N-acetylglucosamine 1236:1254 bisecting N-acetylglucosamine 1226:1254 bisecting N-acetylglucosamine 1226:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 5 120 theme glycan 1198:1203 arg1 determinants 1205:1216 unique glycan determinants 1191:1216 unique glycan determinants such as bisecting N-acetylglucosamine 1191:1254 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 7 121 theme lectin 1566:1571 arg1 data 1592:1595 lectin immunofluorescence data 1566:1595 lectin immunofluorescence data 1566:1595 RNA-Seq and lectin immunofluorescence data correlated well with these data, showing the highest upregulation of Mgat3 and binding with PHA-E in LIM1215. 24295106 8 122 theme Man1α1 1725:1730 arg1 Downregulation 1707:1720 Downregulation 1707:1720 Downregulation of Man1α1 and Mgat1 in LIM1215 1707:1751 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 24295106 5 123 theme high 1275:1278 arg1 level 1280:1284 a high level 1273:1284 a high level in the metastatic LIM1215 cells 1273:1316 Importantly, unique glycan determinants such as bisecting N-acetylglucosamine were observed at a high level in the metastatic LIM1215 cells, with some expressed in the moderately differentiated LIM1899, while none were detected in the poorly differentiated LIM2405 cells. 24295106 8 124 theme Mgat1 1736:1740 arg1 Downregulation 1707:1720 Downregulation 1707:1720 Downregulation of Man1α1 and Mgat1 in LIM1215 1707:1751 Downregulation of Man1α1 and Mgat1 in LIM1215 also coincided with the higher degree of incomplete N-glycan processing and accumulation of high mannose type structures as well as bisecting N-glycans when compared to the other two cell lines. 28244757 3 0 theme azidosugar-labeled 530:547 arg1 glycopeptides 549:561 azidosugar-labeled glycopeptides 530:561 azidosugar-labeled glycopeptides bearing fully intact glycans 530:590 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 1 1 theme glycan 256:261 arg1 structures 263:272 glycan structures 256:272 glycan structures 256:272 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 0 2 theme Intact 74:79 arg1 N- 81:82 Intact N- 74:82 Intact N- 74:82 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 7 3 theme permethylation 1198:1211 arg1 analysis 1213:1220 permethylation analysis 1198:1220 permethylation analysis 1198:1220 Glycopeptide-bound glycans observed by IsoTaG were found to be comparable to released N-glycans identified by permethylation analysis. 28244757 4 4 theme broad 615:619 arg1 application 621:631 the broad application 611:631 the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines 611:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 5 5 gly glycopeptide 789:800 arg2 glycopeptide 789:800 a new high-fidelity pattern-searching and glycopeptide validation algorithm 747:821 glycopeptide 789:800 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 6 6 theme discrete 1060:1067 arg1 structures 1076:1085 74 discrete glycan structures 1057:1085 74 discrete glycan structures 1057:1085 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 0 7 theme Whole 109:113 arg1 Proteomes 120:128 Whole Cell Proteomes 109:128 Whole Cell Proteomes 109:128 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 1 8 contain have 157:160 arg2 variety 174:180 an enormous variety 162:180 an enormous variety of biological consequences 162:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 1 8 contain have 157:160 arg1 glycosylation 139:151 Protein glycosylation 131:151 Protein glycosylation 131:151 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 1 8 contain have 157:160 arg2 consequences 196:207 biological consequences 185:207 biological consequences 185:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 2 9 theme structural 285:294 arg1 diversity 296:304 This same structural diversity 275:304 This same structural diversity 275:304 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 1 10 theme enormous 165:172 arg1 variety 174:180 an enormous variety 162:180 an enormous variety of biological consequences 162:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 1 10 theme enormous 165:172 arg1 consequences 196:207 biological consequences 185:207 biological consequences 185:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 5 11 theme validation 802:811 arg1 algorithm 813:821 a new high-fidelity pattern-searching and glycopeptide validation algorithm 747:821 algorithm 813:821 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 6 12 theme cell 929:932 arg1 lines 934:938 15 cell lines 926:938 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz 926:988 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 7 13 theme released 1165:1172 arg1 N-glycans 1174:1182 released N-glycans 1165:1182 released N-glycans identified by permethylation analysis 1165:1220 Glycopeptide-bound glycans observed by IsoTaG were found to be comparable to released N-glycans identified by permethylation analysis. 28244757 6 14 gly O-glycopeptides 1016:1030 arg2 O-glycopeptides 1016:1030 1375 N- and 2159 O-glycopeptides 999:1030 O-glycopeptides 1016:1030 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 2 15 theme same 280:283 arg1 diversity 296:304 This same structural diversity 275:304 This same structural diversity 275:304 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 4 16 theme tissue-diverse 694:707 arg1 lines 714:718 tissue-diverse cell lines 694:718 tissue-diverse cell lines 694:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 3 17 theme intact 577:582 arg1 glycans 584:590 fully intact glycans 571:590 fully intact glycans 571:590 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 3 18 theme isotope-targeted 439:454 arg1 IsoTaG 473:478 IsoTaG 473:478 IsoTaG 473:478 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 3 18 theme isotope-targeted 439:454 arg1 glycoproteomics 456:470 isotope-targeted glycoproteomics 439:470 isotope-targeted glycoproteomics (IsoTaG) 439:479 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 5 19 theme novel 859:863 arg1 isotope 872:878 novel stable isotope 859:878 novel stable isotope probes 859:885 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 1 20 theme biological 185:194 arg1 consequences 196:207 biological consequences 185:207 biological consequences 185:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 5 21 theme stable 865:870 arg1 isotope 872:878 novel stable isotope 859:878 novel stable isotope probes 859:885 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 3 22 gly glycopeptides 549:561 arg2 glycopeptides 549:561 azidosugar-labeled glycopeptides 530:561 azidosugar-labeled glycopeptides bearing fully intact glycans 530:590 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 2 23 theme major 318:322 arg1 challenges 324:333 imposed major challenges 310:333 imposed major challenges 310:333 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 4 24 from collection 680:689 arg1 glycoproteomes 658:671 glycoproteomes 658:671 glycoproteomes from a collection of tissue-diverse cell lines 658:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 1 25 theme consequences 196:207 arg1 variety 174:180 an enormous variety 162:180 an enormous variety of biological consequences 162:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 1 25 theme consequences 196:207 arg1 consequences 196:207 biological consequences 185:207 biological consequences 185:207 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 0 26 theme IsoTaG 15:20 arg1 Development 0:10 Development 0:10 Development of IsoTaG 0:20 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 0 26 theme IsoTaG 15:20 arg1 Technique 50:58 a Chemical Glycoproteomics Technique 23:58 a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes 23:128 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 0 27 theme Cell 115:118 arg1 Proteomes 120:128 Whole Cell Proteomes 109:128 Whole Cell Proteomes 109:128 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 2 28 theme imposed 310:316 arg1 challenges 324:333 imposed major challenges 310:333 imposed major challenges 310:333 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 0 29 theme Chemical 25:32 arg1 Development 0:10 Development 0:10 Development of IsoTaG 0:20 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 0 29 theme Chemical 25:32 arg1 Technique 50:58 a Chemical Glycoproteomics Technique 23:58 a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes 23:128 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 2 30 contain has 306:308 arg1 diversity 296:304 This same structural diversity 275:304 This same structural diversity 275:304 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 2 30 contain has 306:308 arg2 challenges 324:333 imposed major challenges 310:333 imposed major challenges 310:333 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 5 31 theme new 749:751 arg1 pattern-searching 767:783 a new high-fidelity pattern-searching and glycopeptide validation algorithm 747:821 pattern-searching 767:783 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 6 32 theme platform 914:921 arg1 Application 888:898 Application 888:898 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz 888:988 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 4 33 theme lines 714:718 arg1 collection 680:689 a collection 678:689 a collection of tissue-diverse cell lines 678:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 4 34 theme cell 709:712 arg1 lines 714:718 tissue-diverse cell lines 694:718 tissue-diverse cell lines 694:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 5 35 theme high-fidelity 753:765 arg1 pattern-searching 767:783 a new high-fidelity pattern-searching and glycopeptide validation algorithm 747:821 pattern-searching 767:783 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 6 36 theme IsoTaG 907:912 arg1 platform 914:921 the IsoTaG platform 903:921 the IsoTaG platform 903:921 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 6 37 theme glycan 1069:1074 arg1 structures 1076:1085 74 discrete glycan structures 1057:1085 74 discrete glycan structures 1057:1085 Application of the IsoTaG platform to 15 cell lines metabolically labeled with Ac4GalNAz or Ac4ManNAz revealed 1375 N- and 2159 O-glycopeptides, variously modified with 74 discrete glycan structures. 28244757 8 38 theme glycoproteome 1297:1309 arg1 understanding 1276:1288 structural understanding 1265:1288 structural understanding of the glycoproteome 1265:1309 IsoTaG is therefore positioned to enhance structural understanding of the glycoproteome. 28244757 2 39 theme methods 357:363 arg1 development 342:352 the development 338:352 the development of methods to study the intact glycoproteome 338:397 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 0 40 theme Glycoproteomics 34:48 arg1 Development 0:10 Development 0:10 Development of IsoTaG 0:20 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 0 40 theme Glycoproteomics 34:48 arg1 Technique 50:58 a Chemical Glycoproteomics Technique 23:58 a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes 23:128 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 7 41 theme Glycopeptide-bound 1088:1105 arg1 glycans 1107:1113 Glycopeptide-bound glycans 1088:1113 Glycopeptide-bound glycans observed by IsoTaG 1088:1132 Glycopeptide-bound glycans observed by IsoTaG were found to be comparable to released N-glycans identified by permethylation analysis. 28244757 3 42 theme isotope 497:503 arg1 recoding 505:512 isotope recoding 497:512 isotope recoding 497:512 We recently introduced a method termed isotope-targeted glycoproteomics (IsoTaG), which utilizes isotope recoding to characterize azidosugar-labeled glycopeptides bearing fully intact glycans. 28244757 1 43 theme molecular 225:233 arg1 diversity 235:243 the molecular diversity 221:243 the molecular diversity encoded in glycan structures 221:272 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 8 44 theme structural 1265:1274 arg1 understanding 1276:1288 structural understanding 1265:1288 structural understanding of the glycoproteome 1265:1309 IsoTaG is therefore positioned to enhance structural understanding of the glycoproteome. 28244757 1 45 theme Protein 131:137 arg1 glycosylation 139:151 Protein glycosylation 131:151 Protein glycosylation 131:151 Protein glycosylation can have an enormous variety of biological consequences, reflecting the molecular diversity encoded in glycan structures. 28244757 4 46 theme method 640:645 arg1 application 621:631 the broad application 611:631 the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines 611:718 Here, we describe the broad application of the method to analyze glycoproteomes from a collection of tissue-diverse cell lines. 28244757 2 47 theme intact 378:383 arg1 glycoproteome 385:397 the intact glycoproteome 374:397 the intact glycoproteome 374:397 This same structural diversity has imposed major challenges on the development of methods to study the intact glycoproteome. 28244757 0 48 from Proteomes 120:128 arg1 O-Glycopeptides 88:102 O-Glycopeptides 88:102 O-Glycopeptides 88:102 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 0 48 from Proteomes 120:128 arg1 N- 81:82 Intact N- 74:82 Intact N- 74:82 Development of IsoTaG, a Chemical Glycoproteomics Technique for Profiling Intact N- and O-Glycopeptides from Whole Cell Proteomes. 28244757 5 49 theme isotope 872:878 arg1 probes 880:885 novel stable isotope probes 859:885 novel stable isotope probes 859:885 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28244757 5 50 theme glycopeptide 789:800 arg1 algorithm 813:821 a new high-fidelity pattern-searching and glycopeptide validation algorithm 747:821 algorithm 813:821 The effort was enabled by a new high-fidelity pattern-searching and glycopeptide validation algorithm termed IsoStamp v2.0, as well as by novel stable isotope probes. 28382776 3 0 from situation 477:485 arg1 cavity 499:504 the oral cavity 490:504 the oral cavity 490:504 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 4 1 theme cell 580:583 arg1 composition 593:603 cell surface composition 580:603 cell surface composition 580:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 4 2 with mutants 542:548 arg1 defects 569:575 characterized defects 555:575 characterized defects in cell surface composition 555:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 6 3 theme numbers 1200:1206 arg1 exception 1161:1169 the exception 1157:1169 the exception of Campylobacter rectus cell numbers 1157:1206 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 2 4 theme S- 270:271 arg1 layer 274:278 a characteristic cell surface (S-) layer 239:278 a characteristic cell surface (S-) layer modified with a unique O-glycan 239:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 5 5 theme quantitative 863:874 arg1 reaction 903:910 quantitative real-time polymerase chain reaction 863:910 quantitative real-time polymerase chain reaction 863:910 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 6 6 theme rectus 1188:1193 arg1 numbers 1200:1206 Campylobacter rectus cell numbers 1174:1206 Campylobacter rectus cell numbers 1174:1206 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 2 7 theme unique 296:301 arg1 O-glycan 303:310 a unique O-glycan 294:310 a unique O-glycan 294:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 5 8 theme polymerase 886:895 arg1 reaction 903:910 quantitative real-time polymerase chain reaction 863:910 quantitative real-time polymerase chain reaction 863:910 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 5 9 theme colony-forming 832:845 arg1 counts 852:857 colony-forming unit counts 832:857 colony-forming unit counts 832:857 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 7 10 with aggregation 1355:1365 arg1 gingivalis 1386:1395 Porphyromonas gingivalis 1372:1395 Porphyromonas gingivalis 1372:1395 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 2 11 theme surface 261:267 arg1 layer 274:278 a characteristic cell surface (S-) layer 239:278 a characteristic cell surface (S-) layer modified with a unique O-glycan 239:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 4 12 theme select 669:674 arg1 bacteria 681:688 select oral bacteria 669:688 select oral bacteria 669:688 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 5 13 theme glycan 746:751 arg1 influence 695:703 The influence 691:703 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms 691:796 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 2 14 theme characteristic 241:254 arg1 layer 274:278 a characteristic cell surface (S-) layer 239:278 a characteristic cell surface (S-) layer modified with a unique O-glycan 239:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 1 15 theme multispecies 134:145 arg1 biofilms 147:154 subgingival multispecies biofilms 122:154 subgingival multispecies biofilms 122:154 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 6 16 from changes 1035:1041 arg1 surface 1068:1074 the T. forsythia cell surface 1046:1074 the T. forsythia cell surface 1046:1074 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 4 17 theme bacteria 681:688 arg1 species 658:664 nine species 653:664 nine species of select oral bacteria 653:688 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 0 18 theme mutants 68:74 arg1 Behavior 0:7 Behavior 0:7 Behavior of two Tannerella forsythia strains and their cell surface mutants in 0:77 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 3 19 from role 385:388 arg1 formation 401:409 biofilm formation 393:409 biofilm formation 393:409 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 5 20 theme T. forsythia 712:723 arg1 S-layer 725:731 the T. forsythia S-layer 708:731 the T. forsythia S-layer 708:731 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 5 21 theme confocal 980:987 arg1 microscopy 1004:1013 confocal laser scanning microscopy 980:1013 confocal laser scanning microscopy 980:1013 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 0 22 theme strains 37:43 arg1 Behavior 0:7 Behavior 0:7 Behavior of two Tannerella forsythia strains and their cell surface mutants in 0:77 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 5 23 theme scanning 995:1002 arg1 microscopy 1004:1013 confocal laser scanning microscopy 980:1013 confocal laser scanning microscopy 980:1013 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 7 24 theme Porphyromonas 1372:1384 arg1 gingivalis 1386:1395 Porphyromonas gingivalis 1372:1395 Porphyromonas gingivalis 1372:1395 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 0 25 theme surface 60:66 arg1 mutants 68:74 their cell surface mutants 49:74 their cell surface mutants 49:74 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 8 26 theme selective 1414:1422 arg1 role 1424:1427 a selective role 1412:1427 a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm 1412:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 3 27 theme biofilm 393:399 arg1 formation 401:409 biofilm formation 393:409 biofilm formation 393:409 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 5 28 theme in 954:955 arg1 hybridization 962:974 fluorescence in situ hybridization 941:974 fluorescence in situ hybridization 941:974 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 9 29 theme potential 1637:1645 arg1 role 1647:1650 a potential role 1635:1650 a potential role of T. forsythia cell surface structures in the virulence of this species 1635:1723 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 9 30 from role 1647:1650 arg1 virulence 1699:1707 the virulence 1695:1707 the virulence of this species 1695:1723 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 1 31 theme subgingival 122:132 arg1 biofilms 147:154 subgingival multispecies biofilms 122:154 subgingival multispecies biofilms 122:154 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 8 32 with prevalence 1574:1583 arg1 P. gingivalis 1551:1563 P. gingivalis 1551:1563 P. gingivalis 1551:1563 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 0 33 theme forsythia 27:35 arg1 strains 37:43 two Tannerella forsythia strains 12:43 two Tannerella forsythia strains 12:43 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 6 34 theme T. forsythia 1050:1061 arg1 surface 1068:1074 the T. forsythia cell surface 1046:1074 the T. forsythia cell surface 1046:1074 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 0 35 theme cell 55:58 arg1 mutants 68:74 their cell surface mutants 49:74 their cell surface mutants 49:74 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 9 36 theme cell 1668:1671 arg1 structures 1681:1690 T. forsythia cell surface structures 1655:1690 T. forsythia cell surface structures 1655:1690 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 4 37 theme Different 507:515 arg1 strains 530:536 Different T. forsythia strains 507:536 Different T. forsythia strains 507:536 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 3 38 theme biofilm 449:455 arg1 model 457:461 an in vitro multispecies biofilm model 424:461 an in vitro multispecies biofilm model mimicking the situation in the oral cavity 424:504 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 4 39 with strains 530:536 arg1 defects 569:575 characterized defects 555:575 characterized defects in cell surface composition 555:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 3 40 dep in 427:428 arg1 vitro 430:434 vitro 430:434 vitro 430:434 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 6 41 theme consortium 1140:1149 arg1 composition 1108:1118 the quantitative composition 1091:1118 the quantitative composition of the multispecies consortium 1091:1149 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 4 42 theme surface 585:591 arg1 composition 593:603 cell surface composition 580:603 cell surface composition 580:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 8 43 with co-localization 1530:1544 arg1 P. gingivalis 1551:1563 P. gingivalis 1551:1563 P. gingivalis 1551:1563 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 2 44 contain has 235:237 arg1 bacterium 225:233 The bacterium 221:233 The bacterium 221:233 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 2 44 contain has 235:237 arg2 layer 274:278 a characteristic cell surface (S-) layer 239:278 a characteristic cell surface (S-) layer modified with a unique O-glycan 239:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 8 45 from role 1424:1427 arg1 positioning 1478:1488 the positioning 1474:1488 the positioning of this species within the biofilm 1474:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 5 46 from influence 695:703 arg1 composition 770:780 the bacterial composition 756:780 the bacterial composition of the biofilms 756:796 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 4 47 theme characterized 555:567 arg1 defects 569:575 characterized defects 555:575 characterized defects in cell surface composition 555:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 9 48 theme species 1717:1723 arg1 virulence 1699:1707 the virulence 1695:1707 the virulence of this species 1695:1723 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 7 49 from changes 1297:1303 arg1 glycan 1320:1325 the S-layer glycan 1308:1325 the S-layer glycan 1308:1325 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 1 50 theme Tannerella 157:166 arg1 member 112:117 a member 110:117 a member of subgingival multispecies biofilms 110:154 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 1 50 theme Tannerella 157:166 arg1 forsythia 168:176 Tannerella forsythia 157:176 Tannerella forsythia 157:176 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 6 51 theme cell 1195:1198 arg1 numbers 1200:1206 Campylobacter rectus cell numbers 1174:1206 Campylobacter rectus cell numbers 1174:1206 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 7 52 theme bacterial 1253:1261 arg1 agglomeration 1263:1275 the bacterial agglomeration 1249:1275 the bacterial agglomeration 1249:1275 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 5 53 theme real-time 876:884 arg1 reaction 903:910 quantitative real-time polymerase chain reaction 863:910 quantitative real-time polymerase chain reaction 863:910 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 9 54 theme host 1747:1750 arg1 tissues 1752:1758 host tissues 1747:1758 host tissues 1747:1758 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 6 55 theme Campylobacter 1174:1186 arg1 numbers 1200:1206 Campylobacter rectus cell numbers 1174:1206 Campylobacter rectus cell numbers 1174:1206 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 8 56 theme species 1498:1504 arg1 positioning 1478:1488 the positioning 1474:1488 the positioning of this species within the biofilm 1474:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 5 57 dep in 954:955 arg1 situ 957:960 situ 957:960 situ 957:960 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 5 58 theme unit 847:850 arg1 counts 852:857 colony-forming unit counts 832:857 colony-forming unit counts 832:857 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 9 59 theme immune 1768:1773 arg1 system 1775:1780 the immune system 1764:1780 the immune system 1764:1780 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 7 60 theme T. forsythia 1229:1240 arg1 localization 1213:1224 The localization 1209:1224 The localization of T. forsythia within the bacterial agglomeration 1209:1275 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 5 61 theme attached 737:744 arg1 glycan 746:751 attached glycan 737:751 attached glycan 737:751 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 2 62 theme cell 256:259 arg1 layer 274:278 a characteristic cell surface (S-) layer 239:278 a characteristic cell surface (S-) layer modified with a unique O-glycan 239:310 The bacterium has a characteristic cell surface (S-) layer modified with a unique O-glycan. 28382776 5 63 theme chain 897:901 arg1 reaction 903:910 quantitative real-time polymerase chain reaction 863:910 quantitative real-time polymerase chain reaction 863:910 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 1 64 theme biofilms 147:154 arg1 member 112:117 a member 110:117 a member of subgingival multispecies biofilms 110:154 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 1 64 theme biofilms 147:154 arg1 forsythia 168:176 Tannerella forsythia 157:176 Tannerella forsythia 157:176 As a member of subgingival multispecies biofilms, Tannerella forsythia is commonly associated with periodontitis. 28382776 4 65 theme oral 676:679 arg1 bacteria 681:688 select oral bacteria 669:688 select oral bacteria 669:688 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 8 66 with role 1424:1427 arg1 P. gingivalis 1551:1563 P. gingivalis 1551:1563 P. gingivalis 1551:1563 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 0 67 theme oral 92:95 arg1 biofilms 97:104 oral biofilms 92:104 oral biofilms 92:104 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 9 68 theme structures 1681:1690 arg1 role 1647:1650 a potential role 1635:1650 a potential role of T. forsythia cell surface structures in the virulence of this species 1635:1723 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 7 69 theme S-layer 1312:1318 arg1 glycan 1320:1325 the S-layer glycan 1308:1325 the S-layer glycan 1308:1325 The localization of T. forsythia within the bacterial agglomeration varied depending on changes in the S-layer glycan, and this also affected its aggregation with Porphyromonas gingivalis. 28382776 4 70 from defects 569:575 arg1 composition 593:603 cell surface composition 580:603 cell surface composition 580:603 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 5 71 theme laser 989:993 arg1 microscopy 1004:1013 confocal laser scanning microscopy 980:1013 confocal laser scanning microscopy 980:1013 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 6 72 theme quantitative 1095:1106 arg1 composition 1108:1118 the quantitative composition 1091:1118 the quantitative composition of the multispecies consortium 1091:1149 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 8 73 theme C. rectus 1588:1596 arg1 prevalence 1574:1583 the prevalence 1570:1583 the prevalence of C. rectus 1570:1596 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 8 73 theme C. rectus 1588:1596 arg1 role 1424:1427 a selective role 1412:1427 a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm 1412:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 8 73 theme C. rectus 1588:1596 arg1 co-localization 1530:1544 its co-localization 1526:1544 its co-localization with P. gingivalis 1526:1563 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 5 74 theme S-layer 725:731 arg1 influence 695:703 The influence 691:703 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms 691:796 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 5 75 theme biofilms 789:796 arg1 composition 770:780 the bacterial composition 756:780 the bacterial composition of the biofilms 756:796 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 5 76 theme fluorescence 941:952 arg1 hybridization 962:974 fluorescence in situ hybridization 941:974 fluorescence in situ hybridization 941:974 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 0 77 theme Tannerella 16:25 arg1 strains 37:43 two Tannerella forsythia strains 12:43 two Tannerella forsythia strains 12:43 Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. 28382776 3 78 theme multispecies 436:447 arg1 model 457:461 an in vitro multispecies biofilm model 424:461 an in vitro multispecies biofilm model mimicking the situation in the oral cavity 424:504 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 5 79 theme bacterial 760:768 arg1 composition 770:780 the bacterial composition 756:780 the bacterial composition of the biofilms 756:796 The influence of the T. forsythia S-layer and attached glycan on the bacterial composition of the biofilms was analyzed quantitatively using colony-forming unit counts and quantitative real-time polymerase chain reaction, as well as qualitatively by fluorescence in situ hybridization and confocal laser scanning microscopy. 28382776 6 80 theme cell 1063:1066 arg1 surface 1068:1074 the T. forsythia cell surface 1046:1074 the T. forsythia cell surface 1046:1074 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 3 81 theme in 427:428 arg1 model 457:461 an in vitro multispecies biofilm model 424:461 an in vitro multispecies biofilm model mimicking the situation in the oral cavity 424:504 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 28382776 8 82 from prevalence 1574:1583 arg1 positioning 1478:1488 the positioning 1474:1488 the positioning of this species within the biofilm 1474:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 8 83 from co-localization 1530:1544 arg1 positioning 1478:1488 the positioning 1474:1488 the positioning of this species within the biofilm 1474:1523 This suggests a selective role for the glycosylated T. forsythia S-layer in the positioning of this species within the biofilm, its co-localization with P. gingivalis, and the prevalence of C. rectus. 28382776 9 84 theme T. forsythia 1655:1666 arg1 structures 1681:1690 T. forsythia cell surface structures 1655:1690 T. forsythia cell surface structures 1655:1690 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 4 85 theme T. forsythia 517:528 arg1 strains 530:536 Different T. forsythia strains 507:536 Different T. forsythia strains 507:536 Different T. forsythia strains and mutants with characterized defects in cell surface composition were incorporated into the model, together with nine species of select oral bacteria. 28382776 9 86 theme surface 1673:1679 arg1 structures 1681:1690 T. forsythia cell surface structures 1655:1690 T. forsythia cell surface structures 1655:1690 These findings might translate into a potential role of T. forsythia cell surface structures in the virulence of this species when interacting with host tissues and the immune system, from within or beyond the biofilm. 28382776 6 87 theme multispecies 1127:1138 arg1 consortium 1140:1149 the multispecies consortium 1123:1149 the multispecies consortium 1123:1149 This revealed that changes in the T. forsythia cell surface did not affect the quantitative composition of the multispecies consortium, with the exception of Campylobacter rectus cell numbers. 28382776 3 88 theme oral 494:497 arg1 cavity 499:504 the oral cavity 490:504 the oral cavity 490:504 Both the S-layer and the O-glycan were analyzed in this study for their role in biofilm formation by employing an in vitro multispecies biofilm model mimicking the situation in the oral cavity. 27714445 11 0 theme beneficial 1863:1872 arg1 microbiota 1885:1894 beneficial intestinal microbiota 1863:1894 beneficial intestinal microbiota 1863:1894 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 10 1 theme DNA 1521:1523 arg1 experiments 1539:1549 DNA hybridization experiments 1521:1549 DNA hybridization experiments 1521:1549 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 5 2 theme synthesized 870:880 arg1 disaccharides 882:894 the synthesized disaccharides 866:894 the synthesized disaccharides 866:894 Those reactions were performed in a semi-preparative scale, and the synthesized disaccharides were purified. 27714445 9 3 theme casei 1377:1381 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 1 4 theme moieties 248:255 arg1 Lacto-N-biose 140:152 Lacto-N-biose 140:152 Lacto-N-biose (LNB) 140:158 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 4 theme moieties 248:255 arg1 blocks 205:210 major building blocks 190:210 major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa 190:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 4 theme moieties 248:255 arg1 galacto-N-biose 164:178 galacto-N-biose 164:178 galacto-N-biose (GNB) 164:184 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 0 5 from milk 6:9 arg1 species 131:137 Lactobacillus species 117:137 Lactobacillus species 117:137 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 2 6 theme phospho-β-galactosidase 364:386 arg1 GnbG 388:391 the phospho-β-galactosidase GnbG 360:391 the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB 360:471 We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. 27714445 9 7 theme gasseri 1483:1489 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 10 8 theme L. 1641:1642 arg1 strains 1654:1660 all L. rhamnosus strains 1637:1660 all L. rhamnosus strains 1637:1660 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 9 theme disaccharides 1590:1602 arg1 metabolism 1570:1579 the metabolism 1566:1579 the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested 1566:1691 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 7 10 theme molecular 1028:1036 arg1 structures 1038:1047 the molecular structures 1024:1047 the molecular structures 1024:1047 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 11 11 theme milk 1813:1816 arg1 oligosaccharides 1818:1833 human milk oligosaccharides 1807:1833 human milk oligosaccharides 1807:1833 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 1 12 theme glyco-complexes 260:274 arg1 oligosaccharides 220:235 free oligosaccharides 215:235 free oligosaccharides 215:235 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 12 theme glyco-complexes 260:274 arg1 moieties 248:255 glycan moieties 241:255 glycan moieties 241:255 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 0 13 theme prebiotic 94:102 arg1 potential 104:112 their prebiotic potential 88:112 their prebiotic potential 88:112 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 0 14 from synthesis 50:58 arg1 species 131:137 Lactobacillus species 117:137 Lactobacillus species 117:137 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 1 15 from mucosa 319:324 arg1 present 276:282 present 276:282 present 276:282 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 10 16 theme zeae 1673:1676 arg1 strains 1678:1684 all L. zeae strains 1666:1684 all L. zeae strains 1666:1684 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 1 17 theme present 276:282 arg1 glyco-complexes 260:274 glyco-complexes 260:274 glyco-complexes present in human milk and gastrointestinal mucosa 260:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 0 18 theme Lactobacillus 117:129 arg1 species 131:137 Lactobacillus species 117:137 Lactobacillus species 117:137 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 1 19 from milk 293:296 arg1 present 276:282 present 276:282 present 276:282 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 11 20 theme intestinal 1874:1883 arg1 microbiota 1885:1894 beneficial intestinal microbiota 1863:1894 beneficial intestinal microbiota 1863:1894 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 5 21 theme semi-preparative 838:853 arg1 scale 855:859 a semi-preparative scale 836:859 a semi-preparative scale 836:859 Those reactions were performed in a semi-preparative scale, and the synthesized disaccharides were purified. 27714445 9 22 theme Lactobacillus 1384:1396 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 7 23 theme β1,3-glycosidic 1157:1171 arg1 linkages 1173:1180 β1,3-glycosidic linkages 1157:1180 β1,3-glycosidic linkages 1157:1180 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 9 24 theme Lactobacillus 1449:1461 arg1 zeae 1463:1466 Lactobacillus zeae 1449:1466 Lactobacillus zeae 1449:1466 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 8 25 theme prebiotic 1248:1256 arg1 properties 1258:1267 their potential prebiotic properties 1232:1267 their potential prebiotic properties using Lactobacillus species 1232:1295 The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. 27714445 2 26 theme GNB 469:471 arg1 metabolism 447:456 the metabolism 443:456 the metabolism of LNB and GNB 443:471 We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. 27714445 0 27 from lacto- 23:28 arg1 species 131:137 Lactobacillus species 117:137 Lactobacillus species 117:137 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 7 28 theme NMR 997:999 arg1 spectroscopy 1001:1012 NMR spectroscopy 997:1012 NMR spectroscopy 997:1012 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 3 29 theme GNB 571:573 arg1 production 549:558 the production 545:558 the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine 545:624 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 3 29 theme GNB 571:573 arg1 acceptors 629:637 acceptors 629:637 acceptors 629:637 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 0 30 theme Human 0:4 arg1 milk 6:9 Human milk 0:9 Human milk 0:9 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 10 31 theme L. 1670:1671 arg1 strains 1678:1684 all L. zeae strains 1666:1684 all L. zeae strains 1666:1684 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 32 theme hybridization 1525:1537 arg1 experiments 1539:1549 DNA hybridization experiments 1521:1549 DNA hybridization experiments 1521:1549 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 0 33 theme mucosal 15:21 arg1 lacto- 23:28 mucosal lacto- and galacto-N-biose synthesis 15:58 lacto- 23:28 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 1 34 theme human 287:291 arg1 milk 293:296 human milk 287:296 human milk 287:296 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 3 35 theme LNB 563:565 arg1 production 549:558 the production 545:558 the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine 545:624 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 3 35 theme LNB 563:565 arg1 acceptors 629:637 acceptors 629:637 acceptors 629:637 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 1 36 theme major 190:194 arg1 Lacto-N-biose 140:152 Lacto-N-biose 140:152 Lacto-N-biose (LNB) 140:158 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 36 theme major 190:194 arg1 blocks 205:210 major building blocks 190:210 major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa 190:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 36 theme major 190:194 arg1 galacto-N-biose 164:178 galacto-N-biose 164:178 galacto-N-biose (GNB) 164:184 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 7 37 theme carbohydrates 1057:1069 arg1 structures 1038:1047 the molecular structures 1024:1047 the molecular structures 1024:1047 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 9 38 theme L. 1416:1417 arg1 ATCC 1436:1439 L. rhamnosus strain ATCC 53103 1416:1445 L. rhamnosus strain ATCC 53103 1416:1445 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 1 39 from present 276:282 arg1 milk 293:296 human milk 287:296 human milk 287:296 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 39 from present 276:282 arg1 mucosa 319:324 gastrointestinal mucosa 302:324 gastrointestinal mucosa 302:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 11 40 theme microbiota 1885:1894 arg1 enrichment 1849:1858 selective enrichment 1839:1858 selective enrichment of beneficial intestinal microbiota in breast-fed infants 1839:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 11 41 theme oligosaccharides 1818:1833 arg1 role 1799:1802 the putative role 1786:1802 the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants 1786:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 1 42 theme building 196:203 arg1 Lacto-N-biose 140:152 Lacto-N-biose 140:152 Lacto-N-biose (LNB) 140:158 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 42 theme building 196:203 arg1 blocks 205:210 major building blocks 190:210 major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa 190:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 42 theme building 196:203 arg1 galacto-N-biose 164:178 galacto-N-biose 164:178 galacto-N-biose (GNB) 164:184 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 9 43 theme rhamnosus 1419:1427 arg1 ATCC 1436:1439 L. rhamnosus strain ATCC 53103 1416:1445 L. rhamnosus strain ATCC 53103 1416:1445 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 11 44 from enrichment 1849:1858 arg1 infants 1910:1916 breast-fed infants 1899:1916 breast-fed infants 1899:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 6 45 theme maximum 915:921 arg1 yield 923:927 The maximum yield 911:927 The maximum yield obtained for LNB 911:944 The maximum yield obtained for LNB was 10.7 ± 0.2 g/l and for GNB was 10.8 ± 0.3 g/l. 27714445 8 46 theme purified 1187:1194 arg1 sugars 1196:1201 The purified sugars 1183:1201 The purified sugars 1183:1201 The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. 27714445 1 47 theme gastrointestinal 302:317 arg1 mucosa 319:324 gastrointestinal mucosa 302:324 gastrointestinal mucosa 302:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 2 48 theme LNB 461:463 arg1 metabolism 447:456 the metabolism 443:456 the metabolism of LNB and GNB 443:471 We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. 27714445 7 49 theme absence 1079:1085 arg1 structures 1038:1047 the molecular structures 1024:1047 the molecular structures 1024:1047 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 8 50 theme potential 1238:1246 arg1 properties 1258:1267 their potential prebiotic properties 1232:1267 their potential prebiotic properties using Lactobacillus species 1232:1295 The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. 27714445 3 51 with production 549:558 arg1 N-acetylgalactosamine 604:624 N-acetylgalactosamine 604:624 N-acetylgalactosamine 604:624 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 3 51 with production 549:558 arg1 N-acetylglucosamine 580:598 N-acetylglucosamine 580:598 N-acetylglucosamine 580:598 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 9 52 theme strain 1429:1434 arg1 ATCC 1436:1439 L. rhamnosus strain ATCC 53103 1416:1445 L. rhamnosus strain ATCC 53103 1416:1445 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 2 53 from casei 412:416 arg1 GnbG 388:391 the phospho-β-galactosidase GnbG 360:391 the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB 360:471 We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. 27714445 4 54 theme o-nitrophenyl-β-D-galactopyranoside 734:768 arg1 o-nitrophenyl-β-D-galactopyranoside 734:768 o-nitrophenyl-β-D-galactopyranoside 734:768 o-nitrophenyl-β-D-galactopyranoside 734:768 The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside into LNB and GNB, respectively. 27714445 4 54 theme o-nitrophenyl-β-D-galactopyranoside 734:768 arg1 % 729:729 69 ± 4 and 71 ± 1 % 711:729 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside 711:768 The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside into LNB and GNB, respectively. 27714445 0 55 theme galacto-N-biose 34:48 arg1 synthesis 50:58 mucosal lacto- and galacto-N-biose synthesis 15:58 synthesis 50:58 Human milk and mucosal lacto- and galacto-N-biose synthesis by transgalactosylation and their prebiotic potential in Lactobacillus species. 27714445 11 56 theme selective 1839:1847 arg1 enrichment 1849:1858 selective enrichment 1839:1858 selective enrichment of beneficial intestinal microbiota in breast-fed infants 1839:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 11 57 theme putative 1790:1797 arg1 role 1799:1802 the putative role 1786:1802 the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants 1786:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 3 58 theme transglycosylation 501:518 arg1 reactions 520:528 transglycosylation reactions 501:528 transglycosylation reactions 501:528 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 7 59 theme reaction 1090:1097 arg1 byproducts 1099:1108 reaction byproducts 1090:1108 reaction byproducts 1090:1108 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 3 60 used used 488:491 arg2 GnbG 474:477 GnbG 474:477 GnbG 474:477 GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. 27714445 9 61 dep Lactobacillus 1384:1396 arg1 rhamnosus 1398:1406 Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103) 1384:1446 Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103) 1384:1446 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 1 62 theme free 215:218 arg1 oligosaccharides 220:235 free oligosaccharides 215:235 free oligosaccharides 215:235 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 7 63 theme byproducts 1099:1108 arg1 carbohydrates 1057:1069 carbohydrates 1057:1069 carbohydrates 1057:1069 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 7 63 theme byproducts 1099:1108 arg1 absence 1079:1085 the absence 1075:1085 the absence of reaction byproducts 1075:1108 NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. 27714445 11 64 theme human 1807:1811 arg1 oligosaccharides 1818:1833 human milk oligosaccharides 1807:1833 human milk oligosaccharides 1807:1833 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 11 65 theme breast-fed 1899:1908 arg1 infants 1910:1916 breast-fed infants 1899:1916 breast-fed infants 1899:1916 The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants. 27714445 1 66 theme oligosaccharides 220:235 arg1 Lacto-N-biose 140:152 Lacto-N-biose 140:152 Lacto-N-biose (LNB) 140:158 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 66 theme oligosaccharides 220:235 arg1 blocks 205:210 major building blocks 190:210 major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa 190:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 66 theme oligosaccharides 220:235 arg1 galacto-N-biose 164:178 galacto-N-biose 164:178 galacto-N-biose (GNB) 164:184 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 10 67 theme casei 1622:1626 arg1 strains 1628:1634 9 out of 10 L. casei strains 1607:1634 9 out of 10 L. casei strains 1607:1634 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 68 theme homologous 1731:1740 arg1 phospho-β-galactosidase 1707:1729 a phospho-β-galactosidase 1705:1729 a phospho-β-galactosidase homologous to GnbG 1705:1748 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 1 69 attach present 276:282 arg1 milk 293:296 human milk 287:296 human milk 287:296 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 69 attach present 276:282 arg1 mucosa 319:324 gastrointestinal mucosa 302:324 gastrointestinal mucosa 302:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 1 69 attach present 276:282 arg2 glyco-complexes 260:274 glyco-complexes 260:274 glyco-complexes present in human milk and gastrointestinal mucosa 260:324 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 27714445 4 70 theme reaction 658:665 arg1 kinetics 667:674 The reaction kinetics 654:674 The reaction kinetics 654:674 The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-D-galactopyranoside into LNB and GNB, respectively. 27714445 2 71 theme Lactobacillus 398:410 arg1 casei 412:416 Lactobacillus casei BL23 398:421 Lactobacillus casei BL23 398:421 We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. 27714445 9 72 theme johnsonii 1510:1518 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 9 73 theme zeae 1463:1466 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 10 74 theme L. 1619:1620 arg1 strains 1628:1634 9 out of 10 L. casei strains 1607:1634 9 out of 10 L. casei strains 1607:1634 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 9 75 theme tested 1356:1361 arg1 strains 1363:1369 the tested strains 1352:1369 the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii 1352:1518 The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. 27714445 8 76 theme Lactobacillus 1275:1287 arg1 species 1289:1295 Lactobacillus species 1275:1295 Lactobacillus species 1275:1295 The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. 27714445 10 77 theme rhamnosus 1644:1652 arg1 strains 1654:1660 all L. rhamnosus strains 1637:1660 all L. rhamnosus strains 1637:1660 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 78 from metabolism 1570:1579 arg1 strains 1678:1684 all L. zeae strains 1666:1684 all L. zeae strains 1666:1684 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 78 from metabolism 1570:1579 arg1 strains 1654:1660 all L. rhamnosus strains 1637:1660 all L. rhamnosus strains 1637:1660 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 10 78 from metabolism 1570:1579 arg1 strains 1628:1634 9 out of 10 L. casei strains 1607:1634 9 out of 10 L. casei strains 1607:1634 DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. 27714445 1 79 theme glycan 241:246 arg1 moieties 248:255 glycan moieties 241:255 glycan moieties 241:255 Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. 28487369 7 0 link N-linked 1361:1368 arg1 oligosaccharides 1370:1385 N-linked oligosaccharides 1361:1385 N-linked oligosaccharides 1361:1385 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 4 1 with structures 747:756 arg1 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 4 1 with structures 747:756 arg1 ligand 805:810 a preferred galectin ligand 784:810 a preferred galectin ligand 784:810 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 3 2 theme barrier 551:557 arg1 maintenance 521:531 maintenance 521:531 maintenance of the epithelial barrier at the ocular surface 521:579 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 7 3 theme transmembrane 1431:1443 arg1 mucins 1445:1450 transmembrane mucins 1431:1450 transmembrane mucins 1431:1450 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 0 4 theme mucin 72:76 arg1 stability 28:36 stability 28:36 stability 28:36 N-Glycosylation affects the stability and barrier function of the MUC16 mucin. 28487369 0 4 theme mucin 72:76 arg1 function 50:57 barrier function 42:57 barrier function 42:57 N-Glycosylation affects the stability and barrier function of the MUC16 mucin. 28487369 7 5 from stability 1405:1413 arg1 surfaces 1463:1470 mucosal surfaces 1455:1470 mucosal surfaces 1455:1470 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 1 6 from glycocalyx 161:170 arg1 surfaces 183:190 mucosal surfaces 175:190 mucosal surfaces 175:190 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 6 7 theme binding 1207:1213 arg1 affinity 1215:1222 MUC16's binding affinity 1199:1222 MUC16's binding affinity toward galectin-3 1199:1240 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 5 8 from mucin 1058:1062 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 9 theme levels 1036:1041 arg1 loss 1000:1003 partial loss 992:1003 partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 992:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 7 10 theme mucosal 1455:1461 arg1 surfaces 1463:1470 mucosal surfaces 1455:1470 mucosal surfaces 1455:1470 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 2 11 theme N-glycans 296:304 arg1 levels 286:291 Relatively low levels 271:291 Relatively low levels of N-glycans 271:304 Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remain poorly characterized. 28487369 7 12 from function 1419:1426 arg1 surfaces 1463:1470 mucosal surfaces 1455:1470 mucosal surfaces 1455:1470 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 2 13 theme transmembrane 319:331 arg1 mucins 333:338 transmembrane mucins 319:338 transmembrane mucins 319:338 Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remain poorly characterized. 28487369 5 14 with knockdown 915:923 arg1 tunicamycin 884:894 tunicamycin 884:894 tunicamycin 884:894 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 6 15 from retention 1257:1265 arg1 surface 1304:1310 the epithelial cell surface 1284:1310 the epithelial cell surface 1284:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 4 16 theme galectin 796:803 arg1 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 4 16 theme galectin 796:803 arg1 ligand 805:810 a preferred galectin ligand 784:810 a preferred galectin ligand 784:810 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 17 theme N-glycosylation 816:830 arg1 experiments 843:853 N-glycosylation inhibition experiments 816:853 N-glycosylation inhibition experiments 816:853 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 18 theme preferred 786:794 arg1 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 N-acetyllactosamine 763:781 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 4 18 theme preferred 786:794 arg1 ligand 805:810 a preferred galectin ligand 784:810 a preferred galectin ligand 784:810 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 19 theme Golgi 932:936 arg1 MGAT1 978:982 MGAT1 978:982 MGAT1 978:982 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 19 theme Golgi 932:936 arg1 gene 972:975 Golgi N-acetylglucosaminyltransferase I gene 932:975 the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) 928:983 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 20 theme differentiated 670:683 arg1 cells 710:714 differentiated human corneal epithelial cells 670:714 differentiated human corneal epithelial cells 670:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 21 theme MUC16 1065:1069 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 22 from reduction 1090:1098 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 23 theme mucosal 175:181 arg1 surfaces 183:190 mucosal surfaces 175:190 mucosal surfaces 175:190 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 5 24 theme inhibition 832:841 arg1 experiments 843:853 N-glycosylation inhibition experiments 816:853 N-glycosylation inhibition experiments 816:853 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 2 25 theme low 282:284 arg1 levels 286:291 Relatively low levels 271:291 Relatively low levels of N-glycans 271:304 Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remain poorly characterized. 28487369 4 26 theme mucin 651:655 arg1 N-glycans 657:665 transmembrane mucin N-glycans 637:665 transmembrane mucin N-glycans in differentiated human corneal epithelial cells 637:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 3 27 theme mucins 484:489 arg1 interactions 454:465 carbohydrate-dependent interactions 431:465 carbohydrate-dependent interactions of transmembrane mucins with galectin-3 431:505 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 4 28 theme mass 603:606 arg1 spectrometry 608:619 MALDI-TOF mass spectrometry 593:619 MALDI-TOF mass spectrometry 593:619 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 7 29 theme N-linked 1361:1368 arg1 oligosaccharides 1370:1385 N-linked oligosaccharides 1361:1385 N-linked oligosaccharides 1361:1385 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 1 30 theme Transmembrane 79:91 arg1 mucins 93:98 Transmembrane mucins 79:98 Transmembrane mucins 79:98 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 1 30 theme Transmembrane 79:91 arg1 glycoproteins 126:138 highly O-glycosylated glycoproteins 104:138 highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury 104:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 4 31 theme complex-type 734:745 arg1 structures 747:756 complex-type structures 734:756 complex-type structures with N-acetyllactosamine, a preferred galectin ligand 734:810 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 32 theme siRNA-mediated 900:913 arg1 knockdown 915:923 siRNA-mediated knockdown 900:923 siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) 900:983 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 33 theme MALDI-TOF 593:601 arg1 spectrometry 608:619 MALDI-TOF mass spectrometry 593:619 MALDI-TOF mass spectrometry 593:619 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 34 theme gene 972:975 arg1 knockdown 915:923 siRNA-mediated knockdown 900:923 siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) 900:983 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 34 theme gene 972:975 arg1 treatment 869:877 treatment 869:877 treatment with tunicamycin 869:894 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 7 35 theme mucins 1445:1450 arg1 stability 1405:1413 stability 1405:1413 stability 1405:1413 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 7 35 theme mucins 1445:1450 arg1 function 1419:1426 function 1419:1426 function 1419:1426 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 1 36 gly glycoproteins 126:138 arg1 mucins 93:98 Transmembrane mucins 79:98 Transmembrane mucins 79:98 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 1 36 gly glycoproteins 126:138 arg1 glycoproteins 126:138 highly O-glycosylated glycoproteins 104:138 highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury 104:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 5 37 theme cell-surface 1023:1034 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 6 38 dep role 1167:1170 arg1 promoting 1189:1197 promoting 1189:1197 promoting MUC16's binding affinity toward galectin-3 1189:1240 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 6 38 dep role 1167:1170 arg1 causing 1249:1255 causing 1249:1255 causing retention of the lectin on the epithelial cell surface 1249:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 5 39 from function 1122:1129 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 40 theme human 685:689 arg1 cells 710:714 differentiated human corneal epithelial cells 670:714 differentiated human corneal epithelial cells 670:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 41 theme total 1013:1017 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 42 gly O-glycosylated 111:124 arg1 mucins 93:98 Transmembrane mucins 79:98 Transmembrane mucins 79:98 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 1 42 gly O-glycosylated 111:124 arg1 glycoproteins 126:138 highly O-glycosylated glycoproteins 104:138 highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury 104:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 5 43 theme largest 1050:1056 arg1 mucin 1058:1062 the largest mucin 1046:1062 the largest mucin 1046:1062 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 44 theme first 210:214 arg1 line 216:219 the first line 206:219 the first line of cellular defense against infection and injury 206:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 0 45 theme barrier 42:48 arg1 function 50:57 barrier function 42:57 barrier function 42:57 N-Glycosylation affects the stability and barrier function of the MUC16 mucin. 28487369 2 46 located found 310:314 arg1 mucins 333:338 transmembrane mucins 319:338 transmembrane mucins 319:338 Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remain poorly characterized. 28487369 2 46 located found 310:314 arg2 levels 286:291 Relatively low levels 271:291 Relatively low levels of N-glycans 271:304 Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remain poorly characterized. 28487369 5 47 theme N-acetylglucosaminyltransferase 938:968 arg1 MGAT1 978:982 MGAT1 978:982 MGAT1 978:982 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 47 theme N-acetylglucosaminyltransferase 938:968 arg1 gene 972:975 Golgi N-acetylglucosaminyltransferase I gene 932:975 the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) 928:983 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 48 theme epithelial 699:708 arg1 cells 710:714 differentiated human corneal epithelial cells 670:714 differentiated human corneal epithelial cells 670:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 3 49 theme transmembrane 470:482 arg1 mucins 484:489 transmembrane mucins 470:489 transmembrane mucins 470:489 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 3 50 theme carbohydrate-dependent 431:452 arg1 interactions 454:465 carbohydrate-dependent interactions 431:465 carbohydrate-dependent interactions of transmembrane mucins with galectin-3 431:505 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 5 51 theme glycocalyx 1103:1112 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 52 theme I 970:970 arg1 MGAT1 978:982 MGAT1 978:982 MGAT1 978:982 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 52 theme I 970:970 arg1 gene 972:975 Golgi N-acetylglucosaminyltransferase I gene 932:975 the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) 928:983 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 4 53 theme corneal 691:697 arg1 cells 710:714 differentiated human corneal epithelial cells 670:714 differentiated human corneal epithelial cells 670:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 3 54 with interactions 454:465 arg1 galectin-3 496:505 galectin-3 496:505 galectin-3 496:505 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 5 55 theme barrier 1114:1120 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 56 theme O-glycosylated 111:124 arg1 mucins 93:98 Transmembrane mucins 79:98 Transmembrane mucins 79:98 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 1 56 theme O-glycosylated 111:124 arg1 glycoproteins 126:138 highly O-glycosylated glycoproteins 104:138 highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury 104:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 4 57 theme transmembrane 637:649 arg1 N-glycans 657:665 transmembrane mucin N-glycans 637:665 transmembrane mucin N-glycans in differentiated human corneal epithelial cells 637:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 6 58 theme cell 1299:1302 arg1 surface 1304:1310 the epithelial cell surface 1284:1310 the epithelial cell surface 1284:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 3 59 theme epithelial 540:549 arg1 barrier 551:557 the epithelial barrier 536:557 the epithelial barrier 536:557 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 6 60 theme lectin 1274:1279 arg1 retention 1257:1265 retention 1257:1265 retention of the lectin on the epithelial cell surface 1257:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 6 61 theme epithelial 1288:1297 arg1 surface 1304:1310 the epithelial cell surface 1284:1310 the epithelial cell surface 1284:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 0 62 dep stability 28:36 arg1 the 24:26 the 24:26 the 24:26 N-Glycosylation affects the stability and barrier function of the MUC16 mucin. 28487369 7 63 dep stability 1405:1413 arg1 the 1401:1403 the 1401:1403 the 1401:1403 Taken together, these studies define a role for N-linked oligosaccharides in supporting the stability and function of transmembrane mucins on mucosal surfaces. 28487369 1 64 theme cellular 224:231 arg1 defense 233:239 cellular defense 224:239 cellular defense against infection and injury 224:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 5 65 with treatment 869:877 arg1 tunicamycin 884:894 tunicamycin 884:894 tunicamycin 884:894 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 66 theme mucin 1058:1062 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 3 67 theme ocular 566:571 arg1 surface 573:579 the ocular surface 562:579 the ocular surface 562:579 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 28487369 5 68 from levels 1036:1041 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 69 theme defense 233:239 arg1 line 216:219 the first line 206:219 the first line of cellular defense against infection and injury 206:268 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 0 70 theme MUC16 66:70 arg1 mucin 72:76 the MUC16 mucin 62:76 the MUC16 mucin 62:76 N-Glycosylation affects the stability and barrier function of the MUC16 mucin. 28487369 4 71 contain contain 716:722 arg2 structures 747:756 complex-type structures 734:756 complex-type structures with N-acetyllactosamine, a preferred galectin ligand 734:810 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 4 71 contain contain 716:722 arg1 N-glycans 657:665 transmembrane mucin N-glycans 637:665 transmembrane mucin N-glycans in differentiated human corneal epithelial cells 637:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 72 theme concomitant 1078:1088 arg1 reduction 1090:1098 a concomitant reduction 1076:1098 a concomitant reduction in glycocalyx barrier function 1076:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 73 from MUC16 1065:1069 arg1 function 1122:1129 glycocalyx barrier function 1103:1129 glycocalyx barrier function 1103:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 5 74 theme partial 992:998 arg1 loss 1000:1003 partial loss 992:1003 partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 992:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 6 75 theme distinct 1158:1165 arg1 role 1167:1170 a distinct role 1156:1170 a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface 1156:1310 Moreover, we identified a distinct role for N-glycans in promoting MUC16's binding affinity toward galectin-3 and in causing retention of the lectin on the epithelial cell surface. 28487369 4 76 from N-glycans 657:665 arg1 cells 710:714 differentiated human corneal epithelial cells 670:714 differentiated human corneal epithelial cells 670:714 Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand. 28487369 5 77 theme reduction 1090:1098 arg1 levels 1036:1041 both total and cell-surface levels 1008:1041 both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function 1008:1129 In N-glycosylation inhibition experiments, we find that treatment with tunicamycin and siRNA-mediated knockdown of the Golgi N-acetylglucosaminyltransferase I gene (MGAT1) induce partial loss of both total and cell-surface levels of the largest mucin, MUC16, and a concomitant reduction in glycocalyx barrier function. 28487369 1 78 theme apical 154:159 arg1 glycocalyx 161:170 the apical glycocalyx 150:170 the apical glycocalyx on mucosal surfaces 150:190 Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. 28487369 3 79 from surface 573:579 arg1 maintenance 521:531 maintenance 521:531 maintenance of the epithelial barrier at the ocular surface 521:579 We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintenance of the epithelial barrier at the ocular surface. 24607157 5 0 theme glycan 883:888 arg1 units 870:874 native monosaccharide units 848:874 native monosaccharide units of the glycan 848:888 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 1 1 theme carbohydrate 252:263 arg1 structure 265:273 the carbohydrate structure 248:273 the carbohydrate structure determining main function in the cell 248:311 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 2 theme strong 931:936 arg1 correlation 938:948 a strong correlation 929:948 a strong correlation 929:948 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 2 3 theme structure 351:359 arg1 importance 325:334 the importance 321:334 the importance of the primary structure 321:359 Due to the importance of the primary structure, the monosaccharide composition is crucial to show the glycan structure. 24607157 5 4 theme monosaccharide 995:1008 arg1 units 1010:1014 the monosaccharide units 991:1014 the monosaccharide units 991:1014 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 4 5 theme anomeric 726:733 arg1 aldoses 735:741 (1)H/(13)C anomeric aldoses 715:741 (1)H/(13)C anomeric aldoses 715:741 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 4 6 theme 13 721:722 arg1 C 724:724 C 724:724 C 724:724 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 4 7 theme C 724:724 arg1 aldoses 735:741 (1)H/(13)C anomeric aldoses 715:741 (1)H/(13)C anomeric aldoses 715:741 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 3 8 theme similar 535:541 arg1 results 543:549 similar results 535:549 similar results 535:549 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 1 9 theme main 287:290 arg1 function 292:299 main function 287:299 main function in the cell 287:311 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 4 10 theme best 700:703 arg1 value 705:709 the best value 696:709 the best value for (1)H/(13)C anomeric aldoses 696:741 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 4 10 theme best 700:703 arg1 Hz 690:691 155 Hz 686:691 155 Hz 686:691 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 2 11 theme primary 343:349 arg1 structure 351:359 the primary structure 339:359 the primary structure 339:359 Due to the importance of the primary structure, the monosaccharide composition is crucial to show the glycan structure. 24607157 3 12 theme complex 462:468 arg1 carbohydrates 470:482 complex carbohydrates 462:482 complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach 462:634 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 5 13 theme parent 1079:1084 arg1 glycan 1086:1091 the parent glycan 1075:1091 the parent glycan 1075:1091 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 1 14 theme essential 72:80 arg1 functions 82:90 essential functions 72:90 essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction 72:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 15 theme native 848:853 arg1 units 870:874 native monosaccharide units 848:874 native monosaccharide units of the glycan 848:888 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 5 16 theme units 1010:1014 arg1 mobility 979:986 the molecular mobility 965:986 the molecular mobility of the monosaccharide units 965:1014 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 5 17 theme glycan 1086:1091 arg1 properties 1061:1070 the dynamic properties 1049:1070 the dynamic properties of the parent glycan 1049:1091 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 1 18 from function 292:299 arg1 cell 308:311 the cell 304:311 the cell 304:311 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 19 theme monosaccharide 855:868 arg1 units 870:874 native monosaccharide units 848:874 native monosaccharide units of the glycan 848:888 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 0 20 theme Monosaccharide 0:13 arg1 composition 15:25 Monosaccharide composition 0:25 Monosaccharide composition of glycans 0:36 Monosaccharide composition of glycans based on Q-HSQC NMR. 24607157 4 21 theme 1 716:716 arg1 C 724:724 C 724:724 C 724:724 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 1 22 theme related 92:98 arg1 functions 82:90 essential functions 72:90 essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction 72:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 23 theme units 870:874 arg1 quantification 830:843 the quantification 826:843 the quantification of native monosaccharide units of the glycan 826:888 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 0 24 theme glycans 30:36 arg1 composition 15:25 Monosaccharide composition 0:25 Monosaccharide composition of glycans 0:36 Monosaccharide composition of glycans based on Q-HSQC NMR. 24607157 1 25 contain have 67:70 arg1 Glycans 59:65 Glycans 59:65 Glycans 59:65 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 25 contain have 67:70 arg2 functions 82:90 essential functions 72:90 essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction 72:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 26 theme hydrolyzed 909:918 arg1 product 920:926 the hydrolyzed product 905:926 the hydrolyzed product 905:926 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 4 27 theme inverse 795:801 arg1 heads 809:813 conventional inverse probe heads 782:813 conventional inverse probe heads 782:813 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 1 28 theme structural 103:112 arg1 architecture 114:125 structural architecture 103:125 structural architecture 103:125 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 2 29 theme glycan 416:421 arg1 structure 423:431 the glycan structure 412:431 the glycan structure 412:431 Due to the importance of the primary structure, the monosaccharide composition is crucial to show the glycan structure. 24607157 3 30 theme classic 576:582 arg1 approach 627:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 4 31 theme JCH 664:666 arg1 dependence 668:677 JCH dependence 664:677 JCH dependence 664:677 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 5 32 theme dynamic 1053:1059 arg1 properties 1061:1070 the dynamic properties 1049:1070 the dynamic properties of the parent glycan 1049:1091 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 3 33 theme GC-MS-carboxy-reduction/deuterium 584:616 arg1 approach 627:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 1 34 theme cell-cell 220:228 arg1 interaction 230:240 cell-cell interaction 220:240 cell-cell interaction 220:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 5 35 from insights 1037:1044 arg1 properties 1061:1070 the dynamic properties 1049:1070 the dynamic properties of the parent glycan 1049:1091 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 24607157 1 36 theme architecture 114:125 arg1 phenomena 153:161 structural architecture and specific cell surface phenomena 103:161 structural architecture and specific cell surface phenomena 103:161 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 36 theme architecture 114:125 arg1 interaction 230:240 cell-cell interaction 220:240 cell-cell interaction 220:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 36 theme architecture 114:125 arg1 biosignalling 189:201 biosignalling 189:201 biosignalling 189:201 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 36 theme architecture 114:125 arg1 differentiation 172:186 differentiation 172:186 differentiation 172:186 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 36 theme architecture 114:125 arg1 recognition 204:214 recognition 204:214 recognition 204:214 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 0 37 theme Q-HSQC 47:52 arg1 NMR 54:56 Q-HSQC NMR 47:56 Q-HSQC NMR 47:56 Monosaccharide composition of glycans based on Q-HSQC NMR. 24607157 4 38 theme probe 803:807 arg1 heads 809:813 conventional inverse probe heads 782:813 conventional inverse probe heads 782:813 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 3 39 theme NMR 493:495 arg1 spectroscopy 497:508 NMR spectroscopy 493:508 NMR spectroscopy 493:508 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 4 40 dep C 724:724 arg1 H/ 718:719 H/ 718:719 H/ 718:719 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 1 41 theme surface 145:151 arg1 phenomena 153:161 structural architecture and specific cell surface phenomena 103:161 structural architecture and specific cell surface phenomena 103:161 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 41 theme surface 145:151 arg1 interaction 230:240 cell-cell interaction 220:240 cell-cell interaction 220:240 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 41 theme surface 145:151 arg1 biosignalling 189:201 biosignalling 189:201 biosignalling 189:201 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 41 theme surface 145:151 arg1 differentiation 172:186 differentiation 172:186 differentiation 172:186 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 41 theme surface 145:151 arg1 recognition 204:214 recognition 204:214 recognition 204:214 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 4 42 theme milli-microM 753:764 arg1 detection 766:774 milli-microM detection 753:774 milli-microM detection using conventional inverse probe heads 753:813 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 4 43 theme conventional 782:793 arg1 heads 809:813 conventional inverse probe heads 782:813 conventional inverse probe heads 782:813 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 1 44 theme specific 131:138 arg1 surface 145:151 specific cell surface 131:151 specific cell surface 131:151 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 1 45 theme cell 140:143 arg1 surface 145:151 specific cell surface 131:151 specific cell surface 131:151 Glycans have essential functions related to structural architecture and specific cell surface phenomena, such as differentiation, biosignalling, recognition and cell-cell interaction, with the carbohydrate structure determining main function in the cell. 24607157 3 46 theme labeling 618:625 arg1 approach 627:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 the classic GC-MS-carboxy-reduction/deuterium labeling approach 572:634 We now present a method for complex carbohydrates based on NMR spectroscopy, which has shown to give similar results to those obtained by the classic GC-MS-carboxy-reduction/deuterium labeling approach. 24607157 2 47 theme monosaccharide 366:379 arg1 crucial 396:402 crucial 396:402 crucial 396:402 Due to the importance of the primary structure, the monosaccharide composition is crucial to show the glycan structure. 24607157 2 47 theme monosaccharide 366:379 arg1 composition 381:391 the monosaccharide composition 362:391 the monosaccharide composition 362:391 Due to the importance of the primary structure, the monosaccharide composition is crucial to show the glycan structure. 24607157 4 48 theme Quantitative 637:648 arg1 HSQC 650:653 Quantitative HSQC 637:653 Quantitative HSQC 637:653 Quantitative HSQC, through JCH dependence showed 155 Hz as the best value for (1)H/(13)C anomeric aldoses, allowing milli-microM detection using conventional inverse probe heads. 24607157 5 49 theme molecular 969:977 arg1 mobility 979:986 the molecular mobility 965:986 the molecular mobility of the monosaccharide units 965:1014 Combining the quantification of native monosaccharide units of the glycan and those from the hydrolyzed product, a strong correlation occurs between the molecular mobility of the monosaccharide units, giving rise to some insights on the dynamic properties of the parent glycan. 25095792 13 0 dep temperature 2109:2119 arg1 i.e. 2094:2097 i.e. 2094:2097 i.e. 2094:2097 Furthermore, glycosylation protected Cyt c from detrimental effects by some stresses (i.e., elevated temperature and humidity) and from proteolytic degradation. 25095792 9 1 theme ± 1584:1584 arg1 % 1598:1598 92 ± 7% to 96 ± 4% 1581:1598 92 ± 7% to 96 ± 4% within the error 1581:1615 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 2 2 theme physical 353:360 arg1 instability 362:372 their chemical and physical instability 334:372 their chemical and physical instability during production, storage, and delivery 334:413 Although proteins have been increasingly important as pharmaceutical agents, their chemical and physical instability during production, storage, and delivery remains a problem. 25095792 15 3 theme chemical 2384:2391 arg1 method 2433:2438 a potentially valuable method 2410:2438 a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration 2410:2557 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 15 3 theme chemical 2384:2391 arg1 glycosylation 2393:2405 chemical glycosylation 2384:2405 chemical glycosylation 2384:2405 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 14 4 theme important 2301:2309 arg1 glycoconjugates 2284:2298 its glycoconjugates 2280:2298 its glycoconjugates 2280:2298 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 9 5 theme % 1587:1587 arg1 % 1598:1598 92 ± 7% to 96 ± 4% 1581:1598 92 ± 7% to 96 ± 4% within the error 1581:1615 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 1 6 theme BACKGROUND 89:98 arg1 protein 148:154 an apoptosis-initiating protein 124:154 an apoptosis-initiating protein 124:154 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 6 theme BACKGROUND 89:98 arg1 c 111:111 BACKGROUND Cytochrome c 89:111 BACKGROUND Cytochrome c (Cyt c) 89:119 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 6 theme BACKGROUND 89:98 arg1 c 118:118 Cyt c 114:118 Cyt c 114:118 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 7 7 theme c 1041:1041 arg1 glycosylation 1043:1055 Cyt c glycosylation 1037:1055 Cyt c glycosylation 1037:1055 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 14 8 theme non-modified 2182:2193 arg1 c 2199:2199 non-modified Cyt c 2182:2199 non-modified Cyt c 2182:2199 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 3 9 theme long-lasting 539:550 arg1 bioavailability 552:566 their long-lasting bioavailability 533:566 their long-lasting bioavailability 533:566 Chemical glycosylation has been devised as a method to increase protein stability and thus enhance their long-lasting bioavailability. 25095792 5 10 theme 10kD 854:857 arg1 -Cyt-c 859:864 Dex5(10kD)-Cyt-c 849:864 Dex5(10kD)-Cyt-c 849:864 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 14 11 from addition 2172:2179 arg1 susceptible 2210:2220 susceptible 2210:2220 susceptible 2210:2220 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 12 12 theme Cyt 1963:1965 arg1 c 1967:1967 Cyt c 1963:1967 Cyt c 1963:1967 This demonstrates that tertiary structure intactness of Cyt c was essential for apoptosis induction. 25095792 13 13 theme proteolytic 2144:2154 arg1 degradation 2156:2166 proteolytic degradation 2144:2166 proteolytic degradation 2144:2166 Furthermore, glycosylation protected Cyt c from detrimental effects by some stresses (i.e., elevated temperature and humidity) and from proteolytic degradation. 25095792 2 14 theme pharmaceutical 311:324 arg1 agents 326:331 pharmaceutical agents 311:331 pharmaceutical agents 311:331 Although proteins have been increasingly important as pharmaceutical agents, their chemical and physical instability during production, storage, and delivery remains a problem. 25095792 15 15 theme c 2456:2456 arg1 stability 2458:2466 Cyt c stability 2452:2466 Cyt c stability 2452:2466 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 9 16 theme enzyme 1428:1433 arg1 activity 1435:1442 ≥94% residual enzyme activity 1414:1442 ≥94% residual enzyme activity 1414:1442 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 11 17 theme ± 1887:1887 arg1 % 1890:1890 26 ± 6% 1884:1890 26 ± 6% 1884:1890 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 8 18 theme Cyt 1224:1226 arg1 glycoconjugates 1230:1244 the Cyt c glycoconjugates 1220:1244 the Cyt c glycoconjugates 1220:1244 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 9 19 theme Cyt 1521:1523 arg1 activation 1538:1547 non-modified Cyt c. Caspase 9 activation 1508:1547 non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation 1508:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 11 20 theme ± 1875:1875 arg1 % 1878:1878 only 24 ± 8% 1867:1878 only 24 ± 8% 1867:1878 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 1 21 theme eukaryotic 192:201 arg1 cells 203:207 eukaryotic cells 192:207 eukaryotic cells 192:207 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 9 22 theme non-modified 1508:1519 arg1 activation 1538:1547 non-modified Cyt c. Caspase 9 activation 1508:1547 non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation 1508:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 11 23 from drop 1829:1832 arg1 caspase 1837:1843 caspase 3 and 9 activation 1837:1862 caspase 1837:1843 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 23 from drop 1829:1832 arg1 structure 1813:1821 the tertiary structure 1800:1821 the tertiary structure 1800:1821 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 23 from drop 1829:1832 arg1 activation 1853:1862 caspase 3 and 9 activation 1837:1862 activation 1853:1862 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 8 24 theme glycoconjugates 1230:1244 arg1 Functionality 1203:1215 Functionality 1203:1215 Functionality of the Cyt c glycoconjugates 1203:1244 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 7 25 theme CD 1007:1008 arg1 spectra 1011:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 4 26 dep glycans 610:616 arg1 lactose 619:625 lactose 619:625 lactose 619:625 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 26 dep glycans 610:616 arg1 dextrans 635:642 two dextrans 631:642 two dextrans with 1 kD and 10 kD 631:662 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 10 27 theme c 1690:1690 arg1 activity 1692:1699 Cyt c activity 1686:1699 Cyt c activity 1686:1699 There were no major changes in Cyt c activity upon glycosylation. 25095792 5 28 theme Dex5 849:852 arg1 -Cyt-c 859:864 Dex5(10kD)-Cyt-c 849:864 Dex5(10kD)-Cyt-c 849:864 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 4 29 theme c 712:712 arg1 Lys 722:724 Lys 722:724 Lys 722:724 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 29 theme c 712:712 arg1 lysine 714:719 surface exposed Cyt c lysine 692:719 surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds 692:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 30 theme dichroism 996:1004 arg1 spectra 1011:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 9 31 dep showed 1407:1412 arg1 compared 1496:1503 compared 1496:1503 compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation 1496:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 9 32 theme c. 1525:1526 arg1 activation 1538:1547 non-modified Cyt c. Caspase 9 activation 1508:1547 non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation 1508:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 9 33 theme ± 1461:1461 arg1 % 1464:1464 95 ± 1% 1458:1464 95 ± 1% 1458:1464 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 8 34 theme caspase 1285:1291 arg1 assays 1319:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 7 35 theme structure 1178:1186 arg1 perturbations 1188:1200 some minor tertiary structure perturbations 1158:1200 some minor tertiary structure perturbations 1158:1200 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 5 36 theme Dex8 867:870 arg1 -Cyt-c 877:882 Dex8(10kD)-Cyt-c 867:882 Dex8(10kD)-Cyt-c 867:882 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 4 37 theme exposed 700:706 arg1 Lys 722:724 Lys 722:724 Lys 722:724 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 37 theme exposed 700:706 arg1 lysine 714:719 surface exposed Cyt c lysine 692:719 surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds 692:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 38 theme minor 1163:1167 arg1 perturbations 1188:1200 some minor tertiary structure perturbations 1158:1200 some minor tertiary structure perturbations 1158:1200 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 5 39 theme 10kD 872:875 arg1 -Cyt-c 877:882 Dex8(10kD)-Cyt-c 867:882 Dex8(10kD)-Cyt-c 867:882 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 10 40 from changes 1675:1681 arg1 activity 1692:1699 Cyt c activity 1686:1699 Cyt c activity 1686:1699 There were no major changes in Cyt c activity upon glycosylation. 25095792 6 41 theme glycoconjugate 936:949 arg1 structure 951:959 glycoconjugate structure 936:959 glycoconjugate structure 936:959 Subsequently, we investigated glycoconjugate structure, activity, and stability. 25095792 14 42 from formulation 2319:2329 arg1 polymers 2334:2341 polymers 2334:2341 polymers 2334:2341 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 15 43 dep CONCLUSION 2344:2353 arg1 demonstrate 2367:2377 demonstrate 2367:2377 demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration 2367:2557 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 4 44 theme amide 769:773 arg1 bonds 775:779 amide bonds 769:779 amide bonds 769:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 45 theme glycosylation 1130:1142 arg1 levels 1144:1149 high glycosylation levels 1125:1149 high glycosylation levels 1125:1149 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 0 46 theme protein 70:76 arg1 stability 78:86 physical and chemical protein stability 48:86 physical and chemical protein stability 48:86 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 9 47 with same 1621:1624 arg1 % 1598:1598 92 ± 7% to 96 ± 4% 1581:1598 92 ± 7% to 96 ± 4% within the error 1581:1615 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 0 48 theme Chemical 0:7 arg1 glycosylation 9:21 Chemical glycosylation 0:21 Chemical glycosylation of cytochrome c 0:37 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 1 49 theme Cytochrome 100:109 arg1 protein 148:154 an apoptosis-initiating protein 124:154 an apoptosis-initiating protein 124:154 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 49 theme Cytochrome 100:109 arg1 c 111:111 BACKGROUND Cytochrome c 89:111 BACKGROUND Cytochrome c (Cyt c) 89:119 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 49 theme Cytochrome 100:109 arg1 c 118:118 Cyt c 114:118 Cyt c 114:118 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 13 50 theme detrimental 2056:2066 arg1 effects 2068:2074 detrimental effects 2056:2074 detrimental effects 2056:2074 Furthermore, glycosylation protected Cyt c from detrimental effects by some stresses (i.e., elevated temperature and humidity) and from proteolytic degradation. 25095792 9 51 theme Caspase 1528:1534 arg1 activation 1538:1547 non-modified Cyt c. Caspase 9 activation 1508:1547 non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation 1508:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 4 52 theme different 583:591 arg1 glycans 610:616 Three different molecular weight glycans 577:616 Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) 577:663 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 8 53 theme pseudo 1363:1368 arg1 activity 1377:1384 the peroxidase-like pseudo enzyme activity 1343:1384 the peroxidase-like pseudo enzyme activity 1343:1384 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 0 54 theme c 37:37 arg1 glycosylation 9:21 Chemical glycosylation 0:21 Chemical glycosylation of cytochrome c 0:37 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 9 55 dep % 1598:1598 arg1 to 1589:1590 to 1589:1590 to 1589:1590 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 0 56 gly glycosylation 9:21 arg1 c 37:37 cytochrome c 26:37 cytochrome c 26:37 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 0 57 theme chemical 61:68 arg1 stability 78:86 physical and chemical protein stability 48:86 physical and chemical protein stability 48:86 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 14 58 theme solvent 2257:2263 arg1 interface 2265:2273 a water-organic solvent interface 2241:2273 a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers 2241:2341 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 0 59 theme physical 48:55 arg1 stability 78:86 physical and chemical protein stability 48:86 physical and chemical protein stability 48:86 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 12 60 theme apoptosis 1987:1995 arg1 induction 1997:2005 apoptosis induction 1987:2005 apoptosis induction 1987:2005 This demonstrates that tertiary structure intactness of Cyt c was essential for apoptosis induction. 25095792 1 61 theme apoptosis-initiating 127:146 arg1 c 111:111 BACKGROUND Cytochrome c 89:111 BACKGROUND Cytochrome c (Cyt c) 89:119 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 61 theme apoptosis-initiating 127:146 arg1 protein 148:154 an apoptosis-initiating protein 124:154 an apoptosis-initiating protein 124:154 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 11 62 theme -Cyt 1745:1748 arg1 kD 1742:1743 1 kD 1740:1743 1 kD 1740:1743 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 62 theme -Cyt 1745:1748 arg1 c 1750:1750 Dex3(1 kD)-Cyt c 1735:1750 Dex3(1 kD)-Cyt c 1735:1750 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 9 63 theme % 1417:1417 arg1 activity 1435:1442 ≥94% residual enzyme activity 1414:1442 ≥94% residual enzyme activity 1414:1442 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 1 64 theme possible 225:232 arg1 candidate 246:254 a possible cancer drug candidate 223:254 a possible cancer drug candidate 223:254 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 65 theme drug 241:244 arg1 candidate 246:254 a possible cancer drug candidate 223:254 a possible cancer drug candidate 223:254 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 4 66 theme weight 603:608 arg1 glycans 610:616 Three different molecular weight glycans 577:616 Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) 577:663 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 67 theme Cyt 1037:1039 arg1 c 1041:1041 Cyt c 1037:1041 Cyt c glycosylation 1037:1055 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 14 68 theme Cyt 2195:2197 arg1 c 2199:2199 non-modified Cyt c 2182:2199 non-modified Cyt c 2182:2199 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 9 69 theme ≥94 1414:1416 arg1 % 1417:1417 % 1417:1417 % 1417:1417 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 9 70 theme ± 1595:1595 arg1 % 1598:1598 92 ± 7% to 96 ± 4% 1581:1598 92 ± 7% to 96 ± 4% within the error 1581:1615 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 12 71 theme structure 1939:1947 arg1 intactness 1949:1958 tertiary structure intactness 1930:1958 tertiary structure intactness of Cyt c 1930:1967 This demonstrates that tertiary structure intactness of Cyt c was essential for apoptosis induction. 25095792 9 72 theme residual 1419:1426 arg1 activity 1435:1442 ≥94% residual enzyme activity 1414:1442 ≥94% residual enzyme activity 1414:1442 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 10 73 theme major 1669:1673 arg1 changes 1675:1681 no major changes 1666:1681 no major changes in Cyt c activity upon glycosylation 1666:1718 There were no major changes in Cyt c activity upon glycosylation. 25095792 9 74 theme caspase 1475:1481 arg1 activation 1485:1494 95 ± 1% relative caspase 3 activation 1458:1494 95 ± 1% relative caspase 3 activation 1458:1494 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 2 75 theme chemical 340:347 arg1 instability 362:372 their chemical and physical instability 334:372 their chemical and physical instability during production, storage, and delivery 334:413 Although proteins have been increasingly important as pharmaceutical agents, their chemical and physical instability during production, storage, and delivery remains a problem. 25095792 15 76 theme Cyt 2452:2454 arg1 stability 2458:2466 Cyt c stability 2452:2466 Cyt c stability 2452:2466 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 1 77 theme cells 203:207 arg1 cytoplasm 179:187 the cytoplasm 175:187 the cytoplasm of eukaryotic cells 175:207 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 11 78 from loss 1792:1795 arg1 caspase 1837:1843 caspase 3 and 9 activation 1837:1862 caspase 1837:1843 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 78 from loss 1792:1795 arg1 structure 1813:1821 the tertiary structure 1800:1821 the tertiary structure 1800:1821 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 78 from loss 1792:1795 arg1 activation 1853:1862 caspase 3 and 9 activation 1837:1862 activation 1853:1862 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 8 79 theme c 1228:1228 arg1 glycoconjugates 1230:1244 the Cyt c glycoconjugates 1220:1244 the Cyt c glycoconjugates 1220:1244 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 7 80 theme Circular 987:994 arg1 spectra 1011:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra 987:1017 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 10 81 theme Cyt 1686:1688 arg1 activity 1692:1699 Cyt c activity 1686:1699 Cyt c activity 1686:1699 There were no major changes in Cyt c activity upon glycosylation. 25095792 1 82 theme Cyt 114:116 arg1 c 111:111 BACKGROUND Cytochrome c 89:111 BACKGROUND Cytochrome c (Cyt c) 89:119 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 1 82 theme Cyt 114:116 arg1 c 118:118 Cyt c 114:118 Cyt c 114:118 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 25095792 4 83 theme Cyt 708:710 arg1 Lys 722:724 Lys 722:724 Lys 722:724 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 83 theme Cyt 708:710 arg1 lysine 714:719 surface exposed Cyt c lysine 692:719 surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds 692:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 84 theme secondary 1098:1106 arg1 structure 1108:1116 the secondary structure 1094:1116 the secondary structure 1094:1116 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 4 85 with lactose 619:625 arg1 kD 651:652 1 kD 649:652 1 kD 649:652 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 85 with lactose 619:625 arg1 kD 661:662 10 kD 658:662 10 kD 658:662 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 13 86 theme elevated 2100:2107 arg1 temperature 2109:2119 elevated temperature 2100:2119 elevated temperature 2100:2119 Furthermore, glycosylation protected Cyt c from detrimental effects by some stresses (i.e., elevated temperature and humidity) and from proteolytic degradation. 25095792 9 87 theme caspase 1633:1639 arg1 activation 1643:1652 the caspase 3 activation 1629:1652 the caspase 3 activation 1629:1652 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 14 88 from susceptible 2210:2220 arg1 addition 2172:2179 addition 2172:2179 addition 2172:2179 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 4 89 theme lysine 714:719 arg1 residues 727:734 surface exposed Cyt c lysine (Lys) residues 692:734 surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds 692:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 7 90 theme tertiary 1169:1176 arg1 perturbations 1188:1200 some minor tertiary structure perturbations 1158:1200 some minor tertiary structure perturbations 1158:1200 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 8 91 theme cell-free 1275:1283 arg1 assays 1319:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 9 92 theme relative 1466:1473 arg1 activation 1485:1494 95 ± 1% relative caspase 3 activation 1458:1494 95 ± 1% relative caspase 3 activation 1458:1494 The glycoconjugates showed ≥94% residual enzyme activity and 86 ± 3 to 95 ± 1% relative caspase 3 activation compared to non-modified Cyt c. Caspase 9 activation by the glycoconjugates was with 92 ± 7% to 96 ± 4% within the error the same as the caspase 3 activation. 25095792 4 93 dep exposed 700:706 arg1 surface 692:698 surface 692:698 surface 692:698 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 94 theme succinimidyl 742:753 arg1 chemistry 755:763 succinimidyl chemistry 742:763 succinimidyl chemistry 742:763 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 8 95 theme caspase 1299:1305 arg1 assays 1319:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 7 96 theme high 1125:1128 arg1 levels 1144:1149 high glycosylation levels 1125:1149 high glycosylation levels 1125:1149 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 11 97 with Incubation 1721:1730 arg1 mercaptoethanol 1757:1771 mercaptoethanol 1757:1771 mercaptoethanol 1757:1771 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 8 98 theme induction 1309:1317 arg1 assays 1319:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 cell-free caspase 3 and caspase 9 induction assays 1275:1324 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 11 99 theme significant 1780:1790 arg1 loss 1792:1795 significant loss 1780:1795 significant loss in the tertiary structure 1780:1821 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 4 100 dep RESULTS 569:575 arg1 coupled 681:687 coupled 681:687 were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds 665:779 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 5 101 theme Dex3 889:892 arg1 -Cyt-c 898:903 Dex3(1kD)-Cyt-c 889:903 Dex3(1kD)-Cyt-c 889:903 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 3 102 theme Chemical 434:441 arg1 glycosylation 443:455 Chemical glycosylation 434:455 Chemical glycosylation 434:455 Chemical glycosylation has been devised as a method to increase protein stability and thus enhance their long-lasting bioavailability. 25095792 3 102 theme Chemical 434:441 arg1 method 479:484 a method 477:484 a method to increase protein stability and thus enhance their long-lasting bioavailability 477:566 Chemical glycosylation has been devised as a method to increase protein stability and thus enhance their long-lasting bioavailability. 25095792 0 103 theme cytochrome 26:35 arg1 c 37:37 cytochrome c 26:37 cytochrome c 26:37 Chemical glycosylation of cytochrome c improves physical and chemical protein stability. 25095792 5 104 theme 1kD 894:896 arg1 -Cyt-c 898:903 Dex3(1kD)-Cyt-c 889:903 Dex3(1kD)-Cyt-c 889:903 Five neo-glycoconjugates were synthesized, Lac4-Cyt-c, Lac9-Cyt-c, Dex5(10kD)-Cyt-c, Dex8(10kD)-Cyt-c, and Dex3(1kD)-Cyt-c. 25095792 4 105 theme molecular 593:601 arg1 glycans 610:616 Three different molecular weight glycans 577:616 Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) 577:663 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 11 106 theme tertiary 1804:1811 arg1 structure 1813:1821 the tertiary structure 1800:1821 the tertiary structure 1800:1821 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 12 107 theme tertiary 1930:1937 arg1 intactness 1949:1958 tertiary structure intactness 1930:1958 tertiary structure intactness of Cyt c 1930:1967 This demonstrates that tertiary structure intactness of Cyt c was essential for apoptosis induction. 25095792 15 108 theme valuable 2424:2431 arg1 method 2433:2438 a potentially valuable method 2410:2438 a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration 2410:2557 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 15 108 theme valuable 2424:2431 arg1 glycosylation 2393:2405 chemical glycosylation 2384:2405 chemical glycosylation 2384:2405 CONCLUSION The results demonstrate that chemical glycosylation is a potentially valuable method to increase Cyt c stability during formulation and storage and potentially during its application after administration. 25095792 8 109 theme peroxidase-like 1347:1361 arg1 activity 1377:1384 the peroxidase-like pseudo enzyme activity 1343:1384 the peroxidase-like pseudo enzyme activity 1343:1384 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 7 110 theme significant 1071:1081 arg1 changes 1083:1089 significant changes 1071:1089 significant changes 1071:1089 Circular dichroism (CD) spectra demonstrated that Cyt c glycosylation did not cause significant changes to the secondary structure, while high glycosylation levels caused some minor tertiary structure perturbations. 25095792 8 111 theme enzyme 1370:1375 arg1 activity 1377:1384 the peroxidase-like pseudo enzyme activity 1343:1384 the peroxidase-like pseudo enzyme activity 1343:1384 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 14 112 theme water-organic 2243:2255 arg1 interface 2265:2273 a water-organic solvent interface 2241:2273 a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers 2241:2341 In addition, non-modified Cyt c was more susceptible to denaturation by a water-organic solvent interface than its glycoconjugates, important for the formulation in polymers. 25095792 3 113 theme protein 498:504 arg1 stability 506:514 protein stability 498:514 protein stability 498:514 Chemical glycosylation has been devised as a method to increase protein stability and thus enhance their long-lasting bioavailability. 25095792 13 114 theme Cyt 2045:2047 arg1 c 2049:2049 Cyt c 2045:2049 Cyt c 2045:2049 Furthermore, glycosylation protected Cyt c from detrimental effects by some stresses (i.e., elevated temperature and humidity) and from proteolytic degradation. 25095792 4 115 with dextrans 635:642 arg1 kD 651:652 1 kD 649:652 1 kD 649:652 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 4 115 with dextrans 635:642 arg1 kD 661:662 10 kD 658:662 10 kD 658:662 RESULTS Three different molecular weight glycans (lactose and two dextrans with 1 kD and 10 kD) were chemically coupled to surface exposed Cyt c lysine (Lys) residues using succinimidyl chemistry via amide bonds. 25095792 12 116 theme c 1967:1967 arg1 intactness 1949:1958 tertiary structure intactness 1930:1958 tertiary structure intactness of Cyt c 1930:1967 This demonstrates that tertiary structure intactness of Cyt c was essential for apoptosis induction. 25095792 11 117 theme Dex3 1735:1738 arg1 kD 1742:1743 1 kD 1740:1743 1 kD 1740:1743 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 11 117 theme Dex3 1735:1738 arg1 c 1750:1750 Dex3(1 kD)-Cyt c 1735:1750 Dex3(1 kD)-Cyt c 1735:1750 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 8 118 dep determined 1250:1259 arg1 performing 1264:1273 performing 1264:1273 performing cell-free caspase 3 and caspase 9 induction assays 1264:1324 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 8 118 dep determined 1250:1259 arg1 measuring 1333:1341 measuring 1333:1341 measuring the peroxidase-like pseudo enzyme activity 1333:1384 Functionality of the Cyt c glycoconjugates was determined by performing cell-free caspase 3 and caspase 9 induction assays and by measuring the peroxidase-like pseudo enzyme activity. 25095792 11 119 theme c 1750:1750 arg1 Incubation 1721:1730 Incubation 1721:1730 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol 1721:1771 Incubation of Dex3(1 kD)-Cyt c with mercaptoethanol caused significant loss in the tertiary structure and a drop in caspase 3 and 9 activation to only 24 ± 8% and 26 ± 6%, respectively. 25095792 1 120 theme cancer 234:239 arg1 candidate 246:254 a possible cancer drug candidate 223:254 a possible cancer drug candidate 223:254 BACKGROUND Cytochrome c (Cyt c) is an apoptosis-initiating protein when released into the cytoplasm of eukaryotic cells and therefore a possible cancer drug candidate. 24650731 5 0 theme pgl 610:612 arg1 operon 614:619 The pgl operon 606:619 The pgl operon 606:619 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 9 1 theme glycan 1166:1171 arg1 study 1184:1188 Further glycan structural study 1158:1188 Further glycan structural study using the conventional fluorophore-labeling method 1158:1239 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 8 2 theme residue 1149:1155 arg1 presence 1028:1035 the presence 1024:1035 the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1024:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 4 3 theme nucleotide 547:556 arg1 sequence 558:565 overall nucleotide sequence 539:565 overall nucleotide sequence 539:565 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 6 4 theme protein 775:781 arg1 glycosylation 783:795 the protein glycosylation 771:795 the protein glycosylation on the recombinant CmeA 771:819 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 5 5 from CmeA 672:675 arg1 mutant 712:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 9 6 theme fluorophore-labeling 1213:1232 arg1 method 1234:1239 the conventional fluorophore-labeling method 1196:1239 the conventional fluorophore-labeling method 1196:1239 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 2 7 theme C. 327:328 arg1 jejuni 330:335 C. jejuni 327:335 C. jejuni JCM 2013 327:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 11 8 theme prokaryotic 1580:1590 arg1 glycosylation 1592:1604 the observed prokaryotic glycosylation 1567:1604 the observed prokaryotic glycosylation 1567:1604 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 0 9 from 2013 79:82 arg1 operon 42:47 a protein glycosylation operon 18:47 a protein glycosylation operon from Campylobacter jejuni JCM 2013 18:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 0 9 from 2013 79:82 arg1 Identification 0:13 Identification 0:13 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 0:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 0 9 from 2013 79:82 arg1 expression 105:114 its heterologous expression 88:114 its heterologous expression in Escherichia coli 88:134 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 4 10 theme operon 443:448 arg1 similar 467:473 similar 467:473 similar 467:473 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 4 10 theme operon 443:448 arg1 organization 450:461 The operon organization 439:461 The operon organization 439:461 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 8 11 from presence 1028:1035 arg1 end 1087:1089 the reducing end 1074:1089 the reducing end 1074:1089 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 5 12 dep Escherichia 684:694 arg1 coli 696:699 coli 696:699 coli 696:699 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 0 13 from expression 105:114 arg1 2013 79:82 Campylobacter jejuni JCM 2013 54:82 Campylobacter jejuni JCM 2013 54:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 0 13 from expression 105:114 arg1 coli 131:134 Escherichia coli 119:134 Escherichia coli 119:134 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 9 14 theme structural 1173:1182 arg1 study 1184:1188 Further glycan structural study 1158:1188 Further glycan structural study using the conventional fluorophore-labeling method 1158:1239 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 5 15 theme model 658:662 arg1 CmeA 672:675 model protein CmeA 658:675 model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant 658:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 9 16 theme HexNAc-HexNAc-HexNAc-HexNAc 1276:1302 arg1 structure 1304:1312 the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure 1250:1312 the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure 1250:1312 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 7 17 theme mass 958:961 arg1 spectrometry 963:974 mass spectrometry 958:974 mass spectrometry analysis 958:983 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 1 18 theme Campylobacter 137:149 arg1 bacterium 186:194 a human enteropathogenic bacterium 161:194 a human enteropathogenic bacterium possessing an N-glycosylation system 161:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 1 18 theme Campylobacter 137:149 arg1 jejuni 151:156 Campylobacter jejuni 137:156 Campylobacter jejuni 137:156 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 0 19 theme jejuni 68:73 arg1 2013 79:82 Campylobacter jejuni JCM 2013 54:82 Campylobacter jejuni JCM 2013 54:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 2 20 theme protein 250:256 arg1 operon 278:283 a protein glycosylation (pgl) operon 248:283 a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 248:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 8 21 theme spectrometry 995:1006 arg1 analysis 1008:1015 The mass spectrometry analysis 986:1015 The mass spectrometry analysis 986:1015 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 7 22 theme trypsin 928:934 arg1 digestion 936:944 in-gel trypsin digestion 921:944 in-gel trypsin digestion 921:944 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 1 23 theme human 163:167 arg1 bacterium 186:194 a human enteropathogenic bacterium 161:194 a human enteropathogenic bacterium possessing an N-glycosylation system 161:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 1 23 theme human 163:167 arg1 jejuni 151:156 Campylobacter jejuni 137:156 Campylobacter jejuni 137:156 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 0 24 from Identification 0:13 arg1 2013 79:82 Campylobacter jejuni JCM 2013 54:82 Campylobacter jejuni JCM 2013 54:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 0 24 from Identification 0:13 arg1 coli 131:134 Escherichia coli 119:134 Escherichia coli 119:134 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 7 25 dep subjected 908:916 arg1 followed 946:953 followed 946:953 followed by mass spectrometry analysis 946:983 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 10 26 theme UDP-diNAcBac 1352:1363 arg1 synthases 1365:1373 UDP-diNAcBac synthases 1352:1373 UDP-diNAcBac synthases 1352:1373 Transcriptional analysis showed that UDP-diNAcBac synthases and diNAcBac transferase are transcribed but might not function in the constructed system. 24650731 8 27 from end 1087:1089 arg1 residue 1063:1069 the N-acetylhexosamine residue 1040:1069 the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1040:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 8 27 from end 1087:1089 arg1 presence 1028:1035 the presence 1024:1035 the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1024:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 3 28 theme pgl 427:429 arg1 operon 431:436 the pgl operon 423:436 the pgl operon 423:436 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 5 29 theme ΔwaaL 706:710 arg1 mutant 712:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 11 30 theme pgl 1483:1485 arg1 operon 1487:1492 a pgl operon 1481:1492 a pgl operon from C. jejuni JCM 2013 1481:1516 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 6 31 from glycosylation 783:795 arg1 CmeA 816:819 the recombinant CmeA 800:819 the recombinant CmeA 800:819 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 3 32 theme open 382:385 arg1 frames 395:400 Fourteen open reading frames 373:400 Fourteen open reading frames (ORFs) 373:407 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 3 32 theme open 382:385 arg1 ORFs 403:406 ORFs 403:406 ORFs 403:406 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 4 33 theme % 515:515 arg1 identities 525:534 98% and 99% identities 513:534 98% and 99% identities in overall nucleotide sequence and amino acid sequence 513:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 8 34 theme residue 1063:1069 arg1 presence 1028:1035 the presence 1024:1035 the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1024:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 3 35 located found 414:418 arg1 operon 431:436 the pgl operon 423:436 the pgl operon 423:436 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 3 35 located found 414:418 arg2 ORFs 403:406 ORFs 403:406 ORFs 403:406 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 3 35 located found 414:418 arg2 frames 395:400 Fourteen open reading frames 373:400 Fourteen open reading frames (ORFs) 373:407 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 1 36 contain possessing 196:205 arg2 system 226:231 an N-glycosylation system 207:231 an N-glycosylation system 207:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 1 36 contain possessing 196:205 arg1 bacterium 186:194 a human enteropathogenic bacterium 161:194 a human enteropathogenic bacterium possessing an N-glycosylation system 161:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 1 36 contain possessing 196:205 arg1 jejuni 151:156 Campylobacter jejuni 137:156 Campylobacter jejuni 137:156 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 11 37 theme jejuni 1502:1507 arg1 2013 1513:1516 C. jejuni JCM 2013 1499:1516 C. jejuni JCM 2013 1499:1516 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 0 38 theme glycosylation 28:40 arg1 operon 42:47 a protein glycosylation operon 18:47 a protein glycosylation operon from Campylobacter jejuni JCM 2013 18:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 11 39 from 2013 1513:1516 arg1 operon 1487:1492 a pgl operon 1481:1492 a pgl operon from C. jejuni JCM 2013 1481:1516 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 10 40 theme diNAcBac 1379:1386 arg1 transferase 1388:1398 diNAcBac transferase 1379:1398 diNAcBac transferase 1379:1398 Transcriptional analysis showed that UDP-diNAcBac synthases and diNAcBac transferase are transcribed but might not function in the constructed system. 24650731 6 41 theme recombinant 804:814 arg1 CmeA 816:819 the recombinant CmeA 800:819 the recombinant CmeA 800:819 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 8 42 theme predicted 1103:1111 arg1 residue 1149:1155 the predicted di-N-acetylbacillosamine (diNAcBac) residue 1099:1155 the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1040:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 4 43 theme amino 571:575 arg1 sequence 582:589 amino acid sequence 571:589 amino acid sequence 571:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 2 44 gly N-glycosylation 308:322 arg1 2013 341:344 C. jejuni JCM 2013 327:344 C. jejuni JCM 2013 327:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 9 45 theme Further 1158:1164 arg1 study 1184:1188 Further glycan structural study 1158:1188 Further glycan structural study using the conventional fluorophore-labeling method 1158:1239 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 4 46 with similar 467:473 arg1 identities 525:534 98% and 99% identities 513:534 98% and 99% identities in overall nucleotide sequence and amino acid sequence 513:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 10 47 theme constructed 1446:1456 arg1 system 1458:1463 the constructed system 1442:1463 the constructed system 1442:1463 Transcriptional analysis showed that UDP-diNAcBac synthases and diNAcBac transferase are transcribed but might not function in the constructed system. 24650731 2 48 theme prokaryotic 296:306 arg1 N-glycosylation 308:322 prokaryotic N-glycosylation 296:322 prokaryotic N-glycosylation in C. jejuni JCM 2013 296:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 4 49 theme overall 539:545 arg1 sequence 558:565 overall nucleotide sequence 539:565 overall nucleotide sequence 539:565 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 11 50 theme observed 1571:1578 arg1 glycosylation 1592:1604 the observed prokaryotic glycosylation 1567:1604 the observed prokaryotic glycosylation 1567:1604 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 1 51 theme enteropathogenic 169:184 arg1 bacterium 186:194 a human enteropathogenic bacterium 161:194 a human enteropathogenic bacterium possessing an N-glycosylation system 161:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 1 51 theme enteropathogenic 169:184 arg1 jejuni 151:156 Campylobacter jejuni 137:156 Campylobacter jejuni 137:156 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 9 52 theme conventional 1200:1211 arg1 method 1234:1239 the conventional fluorophore-labeling method 1196:1239 the conventional fluorophore-labeling method 1196:1239 Further glycan structural study using the conventional fluorophore-labeling method revealed the GalNAcα-GalNAcα-(Hex-)HexNAc-HexNAc-HexNAc-HexNAc structure. 24650731 2 53 theme jejuni 330:335 arg1 2013 341:344 C. jejuni JCM 2013 327:344 C. jejuni JCM 2013 327:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 8 54 theme mass 990:993 arg1 analysis 1008:1015 The mass spectrometry analysis 986:1015 The mass spectrometry analysis 986:1015 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 6 55 theme lectin-blotting 736:750 arg1 analysis 752:759 The immuno- and lectin-blotting analysis 720:759 The immuno- and lectin-blotting analysis 720:759 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 5 56 theme protein 664:670 arg1 CmeA 672:675 model protein CmeA 658:675 model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant 658:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 1 57 theme N-glycosylation 210:224 arg1 system 226:231 an N-glycosylation system 207:231 an N-glycosylation system 207:231 Campylobacter jejuni is a human enteropathogenic bacterium possessing an N-glycosylation system. 24650731 6 58 gly glycosylation 783:795 arg1 CmeA 816:819 the recombinant CmeA 800:819 the recombinant CmeA 800:819 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 5 59 theme Escherichia 684:694 arg1 mutant 712:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 4 60 theme jejuni 489:494 arg1 11168 501:505 C. jejuni NCTC 11168 486:505 C. jejuni NCTC 11168 486:505 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 2 61 theme pgl 273:275 arg1 operon 278:283 a protein glycosylation (pgl) operon 248:283 a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 248:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 0 62 theme JCM 75:77 arg1 2013 79:82 Campylobacter jejuni JCM 2013 54:82 Campylobacter jejuni JCM 2013 54:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 2 63 theme glycosylation 258:270 arg1 operon 278:283 a protein glycosylation (pgl) operon 248:283 a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 248:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 7 64 theme spectrometry 963:974 arg1 analysis 976:983 mass spectrometry analysis 958:983 mass spectrometry analysis 958:983 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 4 65 theme C. 486:487 arg1 11168 501:505 C. jejuni NCTC 11168 486:505 C. jejuni NCTC 11168 486:505 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 2 66 from N-glycosylation 308:322 arg1 2013 341:344 C. jejuni JCM 2013 327:344 C. jejuni JCM 2013 327:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 4 67 theme NCTC 496:499 arg1 11168 501:505 C. jejuni NCTC 11168 486:505 C. jejuni NCTC 11168 486:505 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 0 68 theme heterologous 92:103 arg1 expression 105:114 its heterologous expression 88:114 its heterologous expression in Escherichia coli 88:134 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 5 69 theme BL21 701:704 arg1 mutant 712:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 the Escherichia coli BL21 ΔwaaL mutant 680:717 The pgl operon was heterologously co-expressed with model protein CmeA in the Escherichia coli BL21 ΔwaaL mutant. 24650731 7 70 theme in-gel 921:926 arg1 trypsin 928:934 in-gel trypsin 921:934 in-gel trypsin digestion 921:944 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 10 71 theme Transcriptional 1315:1329 arg1 analysis 1331:1338 Transcriptional analysis 1315:1338 Transcriptional analysis 1315:1338 Transcriptional analysis showed that UDP-diNAcBac synthases and diNAcBac transferase are transcribed but might not function in the constructed system. 24650731 8 72 theme N-acetylhexosamine 1044:1061 arg1 residue 1063:1069 the N-acetylhexosamine residue 1040:1069 the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1040:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 11 73 theme JCM 1509:1511 arg1 2013 1513:1516 C. jejuni JCM 2013 1499:1516 C. jejuni JCM 2013 1499:1516 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 0 74 theme protein 20:26 arg1 operon 42:47 a protein glycosylation operon 18:47 a protein glycosylation operon from Campylobacter jejuni JCM 2013 18:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 3 75 theme reading 387:393 arg1 frames 395:400 Fourteen open reading frames 373:400 Fourteen open reading frames (ORFs) 373:407 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 3 75 theme reading 387:393 arg1 ORFs 403:406 ORFs 403:406 ORFs 403:406 Fourteen open reading frames (ORFs) were found in the pgl operon. 24650731 4 76 theme % 523:523 arg1 identities 525:534 98% and 99% identities 513:534 98% and 99% identities in overall nucleotide sequence and amino acid sequence 513:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 11 77 theme C. 1499:1500 arg1 2013 1513:1516 C. jejuni JCM 2013 1499:1516 C. jejuni JCM 2013 1499:1516 In conclusion, a pgl operon from C. jejuni JCM 2013 successfully functioned in E. coli, resulting in the observed prokaryotic glycosylation. 24650731 0 78 theme operon 42:47 arg1 Identification 0:13 Identification 0:13 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 0:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 0 78 theme operon 42:47 arg1 expression 105:114 its heterologous expression 88:114 its heterologous expression in Escherichia coli 88:134 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 8 79 theme reducing 1078:1085 arg1 end 1087:1089 the reducing end 1074:1089 the reducing end 1074:1089 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 6 80 theme immuno- 724:730 arg1 analysis 752:759 The immuno- and lectin-blotting analysis 720:759 The immuno- and lectin-blotting analysis 720:759 The immuno- and lectin-blotting analysis indicated the protein glycosylation on the recombinant CmeA. 24650731 4 81 from identities 525:534 arg1 sequence 582:589 amino acid sequence 571:589 amino acid sequence 571:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 4 81 from identities 525:534 arg1 sequence 558:565 overall nucleotide sequence 539:565 overall nucleotide sequence 539:565 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 0 82 theme Campylobacter 54:66 arg1 jejuni 68:73 Campylobacter jejuni 54:73 Campylobacter jejuni JCM 2013 54:82 Identification of a protein glycosylation operon from Campylobacter jejuni JCM 2013 and its heterologous expression in Escherichia coli. 24650731 7 83 theme recombinant 874:884 arg1 CmeA 886:889 the recombinant CmeA 870:889 the recombinant CmeA 870:889 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 24650731 2 84 theme JCM 337:339 arg1 2013 341:344 C. jejuni JCM 2013 327:344 C. jejuni JCM 2013 327:344 In this work, a protein glycosylation (pgl) operon conferring prokaryotic N-glycosylation in C. jejuni JCM 2013 was cloned and identified. 24650731 8 85 theme di-N-acetylbacillosamine 1113:1136 arg1 residue 1149:1155 the predicted di-N-acetylbacillosamine (diNAcBac) residue 1099:1155 the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue 1040:1155 The mass spectrometry analysis showed the presence of the N-acetylhexosamine residue at the reducing end but not the predicted di-N-acetylbacillosamine (diNAcBac) residue. 24650731 4 86 theme acid 577:580 arg1 sequence 582:589 amino acid sequence 571:589 amino acid sequence 571:589 The operon organization was similar to that of C. jejuni NCTC 11168, with 98% and 99% identities in overall nucleotide sequence and amino acid sequence, respectively. 24650731 7 87 theme glycan 850:855 arg1 composition 857:867 the glycan composition 846:867 the glycan composition 846:867 In addition, to analyze the glycan composition, the recombinant CmeA was purified and subjected to in-gel trypsin digestion followed by mass spectrometry analysis. 25464075 2 0 gly glycosylation 312:324 arg1 acceptor 359:366 a readily available lactoside acceptor 329:366 a readily available lactoside acceptor 329:366 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 2 1 theme lacto-N-biose 375:387 arg1 donor 389:393 a lacto-N-biose donor 373:393 a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis 373:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 2 2 with glycosylation 312:324 arg1 donor 389:393 a lacto-N-biose donor 373:393 a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis 373:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 1 3 theme concise 91:97 arg1 synthesis 128:136 A concise and practical chemoenzymatic synthesis 89:136 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) 89:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 2 4 theme efficient 419:427 arg1 synthesis 448:456 a highly efficient one-pot two-enzyme synthesis 410:456 a highly efficient one-pot two-enzyme synthesis 410:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 1 5 theme practical 103:111 arg1 synthesis 128:136 A concise and practical chemoenzymatic synthesis 89:136 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) 89:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 6 theme common 197:202 arg1 structures 209:218 the most common core structures 188:218 the most common core structures of human milk oligosaccharides (HMOs) 188:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 7 theme chemoenzymatic 113:126 arg1 synthesis 128:136 A concise and practical chemoenzymatic synthesis 89:136 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) 89:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 8 theme core 204:207 arg1 structures 209:218 the most common core structures 188:218 the most common core structures of human milk oligosaccharides (HMOs) 188:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 0 9 theme Chemoenzymatic 0:13 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl 0:61 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl lacto-N-tetrasaccharides. 25464075 1 10 theme structures 209:218 arg1 component 167:175 a major component 159:175 a major component 159:175 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 10 theme structures 209:218 arg1 lacto-N-tetraose 141:156 lacto-N-tetraose 141:156 lacto-N-tetraose 141:156 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 0 11 theme lacto-N-tetrasaccharide 28:50 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl 0:61 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl lacto-N-tetrasaccharides. 25464075 1 12 theme lacto-N-tetraose 141:156 arg1 synthesis 128:136 A concise and practical chemoenzymatic synthesis 89:136 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) 89:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 13 theme human 223:227 arg1 HMOs 252:255 HMOs 252:255 HMOs 252:255 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 13 theme human 223:227 arg1 oligosaccharides 234:249 human milk oligosaccharides 223:249 human milk oligosaccharides (HMOs) 223:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 0 14 theme sialyl 56:61 arg1 synthesis 15:23 Chemoenzymatic synthesis 0:23 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl 0:61 Chemoenzymatic synthesis of lacto-N-tetrasaccharide and sialyl lacto-N-tetrasaccharides. 25464075 1 15 theme milk 229:232 arg1 HMOs 252:255 HMOs 252:255 HMOs 252:255 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 15 theme milk 229:232 arg1 oligosaccharides 234:249 human milk oligosaccharides 223:249 human milk oligosaccharides (HMOs) 223:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 2 16 theme two-enzyme 437:446 arg1 synthesis 448:456 a highly efficient one-pot two-enzyme synthesis 410:456 a highly efficient one-pot two-enzyme synthesis 410:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 1 17 theme oligosaccharides 234:249 arg1 structures 209:218 the most common core structures 188:218 the most common core structures of human milk oligosaccharides (HMOs) 188:256 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 2 18 theme acceptor 359:366 arg1 glycosylation 312:324 the glycosylation 308:324 the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis 308:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 2 19 theme convergent 277:286 arg1 synthesis 288:296 This convergent synthesis 272:296 This convergent synthesis 272:296 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 1 20 theme major 161:165 arg1 component 167:175 a major component 159:175 a major component 159:175 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 1 20 theme major 161:165 arg1 lacto-N-tetraose 141:156 lacto-N-tetraose 141:156 lacto-N-tetraose 141:156 A concise and practical chemoenzymatic synthesis of lacto-N-tetraose, a major component and one of the most common core structures of human milk oligosaccharides (HMOs) was reported. 25464075 2 21 theme available 339:347 arg1 acceptor 359:366 a readily available lactoside acceptor 329:366 a readily available lactoside acceptor 329:366 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 2 22 theme lactoside 349:357 arg1 acceptor 359:366 a readily available lactoside acceptor 329:366 a readily available lactoside acceptor 329:366 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 25464075 2 23 theme one-pot 429:435 arg1 synthesis 448:456 a highly efficient one-pot two-enzyme synthesis 410:456 a highly efficient one-pot two-enzyme synthesis 410:456 This convergent synthesis relies on the glycosylation of a readily available lactoside acceptor with a lacto-N-biose donor generated from a highly efficient one-pot two-enzyme synthesis. 29158133 1 0 dep BACKGROUND 123:132 arg1 regulated 192:200 regulated 192:200 regulated 192:200 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 7 1 theme N-deacetylase 1087:1099 arg1 inactive 1112:1119 inactive 1112:1119 inactive 1112:1119 Unlike NDST-1, NDST-4 did not show directional N-sulfotransferase activity while the N-deacetylase domain was inactive. 29158133 7 1 theme N-deacetylase 1087:1099 arg1 domain 1101:1106 the N-deacetylase domain 1083:1106 the N-deacetylase domain 1083:1106 Unlike NDST-1, NDST-4 did not show directional N-sulfotransferase activity while the N-deacetylase domain was inactive. 29158133 5 2 theme rNDST-4 880:886 arg1 specificity 865:875 the substrate specificity 851:875 the substrate specificity of rNDST-4 851:886 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 6 3 theme Full-length 897:907 arg1 rNDST-4 909:915 RESULTS Full-length rNDST-4 889:915 RESULTS Full-length rNDST-4 889:915 RESULTS Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. 29158133 8 4 theme key 1209:1211 arg1 role 1213:1216 the key role 1205:1216 the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis 1205:1287 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 6 5 theme N-deacetylase 931:943 arg1 activity 945:952 obvious N-deacetylase activity 923:952 obvious N-deacetylase activity 923:952 RESULTS Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. 29158133 2 6 theme first 319:323 arg1 enzyme 338:343 the first modification enzyme 315:343 the first modification enzyme in the HS biosynthetic pathway 315:374 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 2 6 theme first 319:323 arg1 N-deacetylase/N-sulfotransferase 272:303 N-deacetylase/N-sulfotransferase 272:303 N-deacetylase/N-sulfotransferase (NDST) 272:310 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 4 7 theme regulatory 665:674 arg1 mechanism 676:684 the regulatory mechanism 661:684 the regulatory mechanism of HS biosynthesis 661:703 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 6 8 theme obvious 923:929 arg1 activity 945:952 obvious N-deacetylase activity 923:952 obvious N-deacetylase activity 923:952 RESULTS Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. 29158133 2 9 theme HS 352:353 arg1 pathway 368:374 the HS biosynthetic pathway 348:374 the HS biosynthetic pathway 348:374 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 8 10 theme Individual 1158:1167 arg1 NDST-4 1169:1174 Individual NDST-4 1158:1174 Individual NDST-4 1158:1174 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 8 11 theme GENERAL 1137:1143 arg1 SIGNIFICANCE 1145:1156 GENERAL SIGNIFICANCE 1137:1156 GENERAL SIGNIFICANCE 1137:1156 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 3 12 theme N-sulfo 381:387 arg1 groups 389:394 The N-sulfo groups 377:394 The N-sulfo groups introduced by NDST 377:413 The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases. 29158133 3 13 theme subsequent 481:490 arg1 processes 492:500 subsequent processes 481:500 subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases 481:551 The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases. 29158133 5 14 theme Highly-purified 714:728 arg1 rNDST-4 750:756 rNDST-4 750:756 rNDST-4 750:756 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 14 theme Highly-purified 714:728 arg1 METHODS 706:712 METHODS 706:712 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides 706:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 14 theme Highly-purified 714:728 arg1 NDST-4 742:747 Highly-purified recombinant NDST-4 714:747 Highly-purified recombinant NDST-4 (rNDST-4) 714:757 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 15 theme substrate 855:863 arg1 specificity 865:875 the substrate specificity 851:875 the substrate specificity of rNDST-4 851:886 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 16 theme recombinant 730:740 arg1 rNDST-4 750:756 rNDST-4 750:756 rNDST-4 750:756 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 16 theme recombinant 730:740 arg1 METHODS 706:712 METHODS 706:712 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides 706:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 16 theme recombinant 730:740 arg1 NDST-4 742:747 Highly-purified recombinant NDST-4 714:747 Highly-purified recombinant NDST-4 (rNDST-4) 714:757 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 6 17 theme RESULTS 889:895 arg1 rNDST-4 909:915 RESULTS Full-length rNDST-4 889:915 RESULTS Full-length rNDST-4 889:915 RESULTS Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. 29158133 3 18 theme susceptibility 463:476 arg1 determination 442:454 determination 442:454 determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases 442:551 The N-sulfo groups introduced by NDST are reportedly involved in determination of the susceptibility to subsequent processes catalyzed by C5-epimerse and 3-O-sulfotransferases. 29158133 0 19 theme sulfate 28:34 arg1 Characterization 0:15 Characterization 0:15 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 0:77 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 4 20 theme isoforms 619:626 arg1 specificities 582:594 the substrate specificities 568:594 the substrate specificities of the four human NDST isoforms 568:626 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 8 21 theme N-S 1241:1243 arg1 domains 1245:1251 N-S domains 1241:1251 N-S domains 1241:1251 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 1 22 theme final 138:142 arg1 structure 144:152 The final structure 134:152 The final structure of heparan sulfate chains 134:178 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 0 23 theme heparan 20:26 arg1 sulfate 28:34 heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 20:77 heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 20:77 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 5 24 theme structurally-defined 786:805 arg1 oligosaccharides 807:822 structurally-defined oligosaccharides 786:822 structurally-defined oligosaccharides 786:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 2 25 theme modification 325:336 arg1 enzyme 338:343 the first modification enzyme 315:343 the first modification enzyme in the HS biosynthetic pathway 315:374 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 2 25 theme modification 325:336 arg1 N-deacetylase/N-sulfotransferase 272:303 N-deacetylase/N-sulfotransferase 272:303 N-deacetylase/N-sulfotransferase (NDST) 272:310 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 5 26 theme oligosaccharides 807:822 arg1 library 775:781 a selective library 763:781 a selective library of structurally-defined oligosaccharides 763:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 26 theme oligosaccharides 807:822 arg1 rNDST-4 750:756 rNDST-4 750:756 rNDST-4 750:756 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 26 theme oligosaccharides 807:822 arg1 METHODS 706:712 METHODS 706:712 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides 706:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 26 theme oligosaccharides 807:822 arg1 NDST-4 742:747 Highly-purified recombinant NDST-4 714:747 Highly-purified recombinant NDST-4 (rNDST-4) 714:757 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 8 27 dep CONCLUSION 1122:1131 arg1 NDST-4 1169:1174 Individual NDST-4 1158:1174 Individual NDST-4 1158:1174 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 0 28 theme N-deacetylase/N-sulfotransferase 36:67 arg1 isoform 69:75 N-deacetylase/N-sulfotransferase isoform 4 36:77 heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 20:77 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 8 29 theme domains 1245:1251 arg1 distribution 1225:1236 the distribution 1221:1236 the distribution of N-S domains and N-Ac domains 1221:1268 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 4 30 theme biosynthesis 692:703 arg1 mechanism 676:684 the regulatory mechanism 661:684 the regulatory mechanism of HS biosynthesis 661:703 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 8 31 from role 1213:1216 arg1 biosynthesis 1276:1287 HS biosynthesis 1273:1287 HS biosynthesis 1273:1287 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 8 31 from role 1213:1216 arg1 distribution 1225:1236 the distribution 1221:1236 the distribution of N-S domains and N-Ac domains 1221:1268 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 8 32 theme domains 1262:1268 arg1 distribution 1225:1236 the distribution 1221:1236 the distribution of N-S domains and N-Ac domains 1221:1268 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 6 33 theme N-sulfotransferase 973:990 arg1 activity 992:999 only N-sulfotransferase activity 968:999 only N-sulfotransferase activity 968:999 RESULTS Full-length rNDST-4 lacks obvious N-deacetylase activity, and displays only N-sulfotransferase activity. 29158133 8 34 theme HS 1273:1274 arg1 biosynthesis 1276:1287 HS biosynthesis 1273:1287 HS biosynthesis 1273:1287 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 5 35 dep METHODS 706:712 arg1 library 775:781 a selective library 763:781 a selective library of structurally-defined oligosaccharides 763:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 35 dep METHODS 706:712 arg1 rNDST-4 750:756 rNDST-4 750:756 rNDST-4 750:756 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 35 dep METHODS 706:712 arg1 METHODS 706:712 METHODS 706:712 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides 706:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 35 dep METHODS 706:712 arg1 NDST-4 742:747 Highly-purified recombinant NDST-4 714:747 Highly-purified recombinant NDST-4 (rNDST-4) 714:757 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 4 36 theme NDST 614:617 arg1 isoforms 619:626 the four human NDST isoforms 599:626 the four human NDST isoforms 599:626 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 8 37 theme N-Ac 1257:1260 arg1 domains 1262:1268 N-Ac domains 1257:1268 N-Ac domains 1257:1268 CONCLUSION AND GENERAL SIGNIFICANCE Individual NDST-4 could not effectively assume the key role in the distribution of N-S domains and N-Ac domains in HS biosynthesis in vivo. 29158133 1 38 theme heparan 157:163 arg1 chains 173:178 heparan sulfate chains 157:178 heparan sulfate chains 157:178 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 7 39 theme directional 1037:1047 arg1 activity 1068:1075 directional N-sulfotransferase activity 1037:1075 directional N-sulfotransferase activity 1037:1075 Unlike NDST-1, NDST-4 did not show directional N-sulfotransferase activity while the N-deacetylase domain was inactive. 29158133 4 40 theme human 608:612 arg1 isoforms 619:626 the four human NDST isoforms 599:626 the four human NDST isoforms 599:626 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 5 41 theme selective 765:773 arg1 library 775:781 a selective library 763:781 a selective library of structurally-defined oligosaccharides 763:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 5 41 theme selective 765:773 arg1 METHODS 706:712 METHODS 706:712 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides 706:822 METHODS Highly-purified recombinant NDST-4 (rNDST-4) and a selective library of structurally-defined oligosaccharides were employed to determine the substrate specificity of rNDST-4. 29158133 0 42 dep sulfate 28:34 arg1 isoform 69:75 N-deacetylase/N-sulfotransferase isoform 4 36:77 heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 20:77 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 1 43 theme sulfate 165:171 arg1 chains 173:178 heparan sulfate chains 157:178 heparan sulfate chains 157:178 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 7 44 theme N-sulfotransferase 1049:1066 arg1 activity 1068:1075 directional N-sulfotransferase activity 1037:1075 directional N-sulfotransferase activity 1037:1075 Unlike NDST-1, NDST-4 did not show directional N-sulfotransferase activity while the N-deacetylase domain was inactive. 29158133 1 45 theme chains 173:178 arg1 structure 144:152 The final structure 134:152 The final structure of heparan sulfate chains 134:178 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 2 46 theme biosynthetic 355:366 arg1 pathway 368:374 the HS biosynthetic pathway 348:374 the HS biosynthetic pathway 348:374 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 0 47 theme oligosaccharide 95:109 arg1 substrates 111:120 synthetic oligosaccharide substrates 85:120 synthetic oligosaccharide substrates 85:120 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 1 48 theme template 254:261 arg1 process 263:269 a template process 252:269 a template process 252:269 BACKGROUND The final structure of heparan sulfate chains is strictly regulated in vivo, though the biosynthesis is not guided by a template process. 29158133 0 49 theme synthetic 85:93 arg1 substrates 111:120 synthetic oligosaccharide substrates 85:120 synthetic oligosaccharide substrates 85:120 Characterization of heparan sulfate N-deacetylase/N-sulfotransferase isoform 4 using synthetic oligosaccharide substrates. 29158133 4 50 theme substrate 572:580 arg1 specificities 582:594 the substrate specificities 568:594 the substrate specificities of the four human NDST isoforms 568:626 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 29158133 2 51 from enzyme 338:343 arg1 pathway 368:374 the HS biosynthetic pathway 348:374 the HS biosynthetic pathway 348:374 N-deacetylase/N-sulfotransferase (NDST) is the first modification enzyme in the HS biosynthetic pathway. 29158133 4 52 theme HS 689:690 arg1 biosynthesis 692:703 HS biosynthesis 689:703 HS biosynthesis 689:703 Understanding the substrate specificities of the four human NDST isoforms has become central to uncovering the regulatory mechanism of HS biosynthesis. 24233973 7 0 contain carrying 1699:1706 arg1 protein 1691:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein 1637:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1637:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 0 contain carrying 1699:1706 arg2 Siaα2-3Gal 1708:1717 Siaα2-3Gal 1708:1717 Siaα2-3Gal 1708:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 4 1 theme mass 1062:1065 arg1 spectrometry 1067:1078 mass spectrometry 1062:1078 mass spectrometry 1062:1078 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 3 2 theme embryonic 708:716 arg1 HEK 726:728 HEK 726:728 HEK 726:728 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 3 2 theme embryonic 708:716 arg1 kidney 718:723 Human embryonic kidney 702:723 Human embryonic kidney (HEK) 702:729 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 3 theme multiple 252:259 arg1 copies 261:266 multiple copies 252:266 multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal 252:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 4 theme cell 735:738 arg1 lines 740:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 7 5 theme clone 1457:1461 arg1 lactosamine 1444:1454 lactosamine 1444:1454 lactosamine (clone 293-P) 1444:1468 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 5 theme clone 1457:1461 arg1 293-P 1463:1467 clone 293-P 1457:1467 clone 293-P 1457:1467 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 6 theme sialylated 1411:1420 arg1 core 1422:1425 sialylated core 1 1411:1427 sialylated core 1 1411:1427 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 1 7 theme glycoprotein 176:187 arg1 PSGL-1/mIgG 211:221 PSGL-1/mIgG 211:221 PSGL-1/mIgG(2b) 211:225 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 1 7 theme glycoprotein 176:187 arg1 IgG2b 204:208 P-selectin glycoprotein ligand-1/mouse IgG2b 165:208 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 8 theme lactosamine 1444:1454 arg1 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 6 9 theme multivalent 1264:1274 arg1 binding 1276:1282 a multivalent binding 1262:1282 a multivalent binding 1262:1282 The binding curves displayed a slow dissociation indicating a multivalent binding. 24233973 3 10 theme 293-P 613:617 arg1 lines 624:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 11 theme IgG2b 204:208 arg1 protein 235:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein 165:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 12 gly sialylated 1369:1378 arg1 core 1380:1383 mostly sialylated core 1 1362:1385 mostly sialylated core 1 (clone C-P55) 1362:1399 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 12 gly sialylated 1369:1378 arg1 C-P55 1394:1398 clone C-P55 1388:1398 clone C-P55 1388:1398 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 13 theme large 1641:1645 arg1 PSGL-1/mIgG 1657:1667 the large, flexible PSGL-1/mIgG 1637:1667 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1637:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 13 theme large 1641:1645 arg1 2b 1669:1670 2b 1669:1670 2b 1669:1670 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 14 theme clone 1388:1392 arg1 core 1380:1383 mostly sialylated core 1 1362:1385 mostly sialylated core 1 (clone C-P55) 1362:1399 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 14 theme clone 1388:1392 arg1 C-P55 1394:1398 clone C-P55 1388:1398 clone C-P55 1388:1398 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 3 15 theme TheC-P55 600:607 arg1 lines 624:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 2 16 theme gel 569:571 arg1 chromatography 584:597 gel filtration chromatography 569:597 gel filtration chromatography 569:597 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 0 17 from acid 67:70 arg1 structures 99:108 different O-linked core structures 75:108 different O-linked core structures on mucin-type fusion proteins 75:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 0 18 theme fusion 124:129 arg1 proteins 131:138 mucin-type fusion proteins 113:138 mucin-type fusion proteins 113:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 7 19 theme potential 1620:1628 arg1 use 1630:1632 these structures.The potential use 1599:1632 these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1599:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 1 20 theme influenza 275:283 arg1 Siaα2-3Gal 308:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 21 theme PSGL-1/mIgG 773:783 arg1 protein 796:802 the PSGL-1/mIgG(2b) fusion protein 769:802 the PSGL-1/mIgG(2b) fusion protein 769:802 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 22 theme Biacore 441:447 arg1 biosensor 449:457 a Biacore biosensor 439:457 a Biacore biosensor 439:457 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 23 contain carrying 1353:1360 arg1 PSGL-1/mIgG 1337:1347 PSGL-1/mIgG 1337:1347 PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1337:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 23 contain carrying 1353:1360 arg1 2b 1349:1350 2b 1349:1350 2b 1349:1350 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 23 contain carrying 1353:1360 arg2 core 1380:1383 mostly sialylated core 1 1362:1385 mostly sialylated core 1 (clone C-P55) 1362:1399 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 23 contain carrying 1353:1360 arg2 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 23 contain carrying 1353:1360 arg2 mix 1404:1406 a mix 1402:1406 a mix of sialylated core 1 1402:1427 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 23 contain carrying 1353:1360 arg2 C-P55 1394:1398 clone C-P55 1388:1398 clone C-P55 1388:1398 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 3 24 theme Chinese 667:673 arg1 CHO 690:692 CHO 690:692 CHO 690:692 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 3 24 theme Chinese 667:673 arg1 ovary 683:687 the Chinese hamster ovary 663:687 the Chinese hamster ovary (CHO)-K1 663:696 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 3 25 theme fusion 789:794 arg1 protein 796:802 the PSGL-1/mIgG(2b) fusion protein 769:802 the PSGL-1/mIgG(2b) fusion protein 769:802 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 26 contain carrying 243:250 arg1 protein 235:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein 165:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 1 26 contain carrying 243:250 arg2 copies 261:266 multiple copies 252:266 multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal 252:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 2 27 theme large 516:520 arg1 cultures 528:535 large scale cultures 516:535 large scale cultures 516:535 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 3 28 theme cell 823:826 arg1 line 828:831 the C-PSLex cell line 811:831 the C-PSLex cell line 811:831 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 29 theme recombinant 352:362 arg1 hemagglutinin 403:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 30 theme CHO-K1 864:869 arg1 cells 871:875 CHO-K1 cells 864:875 CHO-K1 cells 864:875 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 0 31 theme different 75:83 arg1 structures 99:108 different O-linked core structures 75:108 different O-linked core structures on mucin-type fusion proteins 75:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 1 32 theme influenza 370:378 arg1 hemagglutinin 403:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 33 theme mucin-type 1673:1682 arg1 protein 1691:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein 1637:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1637:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 0 34 theme core 94:97 arg1 structures 99:108 different O-linked core structures 75:108 different O-linked core structures on mucin-type fusion proteins 75:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 1 35 theme A/Vietnam/1203/04 385:401 arg1 hemagglutinin 403:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 4 36 theme liquid 1038:1043 arg1 chromatography 1045:1058 liquid chromatography 1038:1058 liquid chromatography 1038:1058 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 0 37 theme Avian 0:4 arg1 hemagglutinin 19:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 7 38 theme PSGL-1/mIgG 1657:1667 arg1 protein 1691:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein 1637:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1637:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 0 39 theme H5 16:17 arg1 hemagglutinin 19:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 1 40 theme hemagglutinin 285:297 arg1 Siaα2-3Gal 308:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 41 theme multivalent 1724:1734 arg1 inhibitor 1736:1744 a multivalent inhibitor 1722:1744 a multivalent inhibitor of influenza virus 1722:1763 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 1 42 theme Siaα2-3Gal 308:317 arg1 copies 261:266 multiple copies 252:266 multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal 252:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 4 43 theme proteins 1025:1032 arg1 blotting 1009:1016 lectin Western blotting 994:1016 lectin Western blotting of the proteins 994:1032 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 4 43 theme proteins 1025:1032 arg1 chromatography 1045:1058 liquid chromatography 1038:1058 liquid chromatography 1038:1058 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 4 43 theme proteins 1025:1032 arg1 spectrometry 1067:1078 mass spectrometry 1062:1078 mass spectrometry 1062:1078 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 3 44 theme ovary 683:687 arg1 lines 740:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 0 45 theme high 44:47 arg1 avidity 49:55 high avidity 44:55 high avidity 44:55 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 5 46 theme binding 1179:1185 arg1 partner 1187:1193 a good binding partner 1172:1193 a good binding partner of H5 1172:1199 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 5 46 theme binding 1179:1185 arg1 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG(2b) 1153:1167 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 4 47 theme O-glycans 1108:1116 arg1 blotting 1009:1016 lectin Western blotting 994:1016 lectin Western blotting of the proteins 994:1032 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 4 47 theme O-glycans 1108:1116 arg1 chromatography 1045:1058 liquid chromatography 1038:1058 liquid chromatography 1038:1058 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 4 47 theme O-glycans 1108:1116 arg1 spectrometry 1067:1078 mass spectrometry 1062:1078 mass spectrometry 1062:1078 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 7 48 theme sialylated 1369:1378 arg1 core 1380:1383 mostly sialylated core 1 1362:1385 mostly sialylated core 1 (clone C-P55) 1362:1399 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 48 theme sialylated 1369:1378 arg1 C-P55 1394:1398 clone C-P55 1388:1398 clone C-P55 1388:1398 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 3 49 theme Human 702:706 arg1 HEK 726:728 HEK 726:728 HEK 726:728 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 3 49 theme Human 702:706 arg1 kidney 718:723 Human embryonic kidney 702:723 Human embryonic kidney (HEK) 702:729 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 7 50 theme sialylated 1480:1489 arg1 lactosamine 1491:1501 mainly sialylated lactosamine 1473:1501 mainly sialylated lactosamine (clone C-PSLex) 1473:1517 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 50 theme sialylated 1480:1489 arg1 C-PSLex 1510:1516 clone C-PSLex 1504:1516 clone C-PSLex 1504:1516 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 1 51 theme fusion 228:233 arg1 protein 235:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein 165:241 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 2 52 theme stable 495:500 arg1 lines 507:511 stable cell lines 495:511 stable cell lines in large scale cultures 495:535 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 0 53 link O-linked 85:92 arg1 structures 99:108 different O-linked core structures 75:108 different O-linked core structures on mucin-type fusion proteins 75:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 4 54 theme Western 1001:1007 arg1 blotting 1009:1016 lectin Western blotting 994:1016 lectin Western blotting of the proteins 994:1032 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 1 55 gly glycoprotein 176:187 arg1 glycoprotein 176:187 P-selectin glycoprotein ligand-1/mouse IgG2b 165:208 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 4 56 theme released 1083:1090 arg1 O-glycans 1108:1116 released non-derivatized O-glycans 1083:1116 released non-derivatized O-glycans 1083:1116 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 3 57 theme kidney 718:723 arg1 lines 740:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines 663:744 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 58 theme P-selectin 165:174 arg1 PSGL-1/mIgG 211:221 PSGL-1/mIgG 211:221 PSGL-1/mIgG(2b) 211:225 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 1 58 theme P-selectin 165:174 arg1 IgG2b 204:208 P-selectin glycoprotein ligand-1/mouse IgG2b 165:208 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 59 gly sialylated 1411:1420 arg1 core 1422:1425 sialylated core 1 1411:1427 sialylated core 1 1411:1427 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 60 theme sialylated 1433:1442 arg1 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 61 theme core 1422:1425 arg1 core 1380:1383 mostly sialylated core 1 1362:1385 mostly sialylated core 1 (clone C-P55) 1362:1399 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 61 theme core 1422:1425 arg1 mix 1404:1406 a mix 1402:1406 a mix of sialylated core 1 1402:1427 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 61 theme core 1422:1425 arg1 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 61 theme core 1422:1425 arg1 C-P55 1394:1398 clone C-P55 1388:1398 clone C-P55 1388:1398 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 1 62 theme different 322:330 arg1 chains 341:346 different O-glycan chains 322:346 different O-glycan chains 322:346 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 1 63 theme ligand-1/mouse 189:202 arg1 PSGL-1/mIgG 211:221 PSGL-1/mIgG 211:221 PSGL-1/mIgG(2b) 211:225 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 1 63 theme ligand-1/mouse 189:202 arg1 IgG2b 204:208 P-selectin glycoprotein ligand-1/mouse IgG2b 165:208 P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 165:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 64 theme influenza 1749:1757 arg1 virus 1759:1763 influenza virus 1749:1763 influenza virus 1749:1763 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 0 65 theme mucin-type 113:122 arg1 proteins 131:138 mucin-type fusion proteins 113:138 mucin-type fusion proteins 113:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 2 66 theme filtration 573:582 arg1 chromatography 584:597 gel filtration chromatography 569:597 gel filtration chromatography 569:597 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 7 67 theme protein 1691:1697 arg1 use 1630:1632 these structures.The potential use 1599:1632 these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1599:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 68 theme structures.The 1605:1618 arg1 use 1630:1632 these structures.The potential use 1599:1632 these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1599:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 69 theme fusion 1684:1689 arg1 protein 1691:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein 1637:1697 the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal 1637:1717 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 5 70 theme Biacore 1119:1125 arg1 experiments 1127:1137 Biacore experiments 1119:1137 Biacore experiments 1119:1137 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 3 71 theme cell 619:622 arg1 lines 624:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines 600:628 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 7 72 dep large 1641:1645 arg1 flexible 1648:1655 flexible 1648:1655 flexible 1648:1655 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 0 73 from structures 99:108 arg1 proteins 131:138 mucin-type fusion proteins 113:138 mucin-type fusion proteins 113:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 6 74 theme slow 1233:1236 arg1 dissociation 1238:1249 a slow dissociation 1231:1249 a slow dissociation indicating a multivalent binding 1231:1282 The binding curves displayed a slow dissociation indicating a multivalent binding. 24233973 7 75 theme virus 1759:1763 arg1 inhibitor 1736:1744 a multivalent inhibitor 1722:1744 a multivalent inhibitor of influenza virus 1722:1763 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 0 76 theme sialic 60:65 arg1 acid 67:70 sialic acid 60:70 sialic acid on different O-linked core structures on mucin-type fusion proteins 60:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 7 77 gly sialylated 1480:1489 arg1 lactosamine 1491:1501 mainly sialylated lactosamine 1473:1501 mainly sialylated lactosamine (clone C-PSLex) 1473:1517 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 77 gly sialylated 1480:1489 arg1 C-PSLex 1510:1516 clone C-PSLex 1504:1516 clone C-PSLex 1504:1516 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 2 78 theme scale 522:526 arg1 cultures 528:535 large scale cultures 516:535 large scale cultures 516:535 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 6 79 theme binding 1206:1212 arg1 curves 1214:1219 The binding curves 1202:1219 The binding curves 1202:1219 The binding curves displayed a slow dissociation indicating a multivalent binding. 24233973 4 80 theme lectin 994:999 arg1 blotting 1009:1016 lectin Western blotting 994:1016 lectin Western blotting of the proteins 994:1032 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 0 81 theme O-linked 85:92 arg1 structures 99:108 different O-linked core structures 75:108 different O-linked core structures on mucin-type fusion proteins 75:138 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 1 82 theme human 364:368 arg1 hemagglutinin 403:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 83 theme C-PSLex 815:821 arg1 line 828:831 the C-PSLex cell line 811:831 the C-PSLex cell line 811:831 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 1 84 theme H5N1 380:383 arg1 hemagglutinin 403:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin 352:415 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 85 theme core 908:911 arg1 β1,6GnT-I 915:923 the core 2 β1,6GnT-I 904:923 the core 2 β1,6GnT-I 904:923 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 2 86 from lines 507:511 arg1 cultures 528:535 large scale cultures 516:535 large scale cultures 516:535 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 0 87 theme influenza 6:14 arg1 hemagglutinin 19:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin 0:31 Avian influenza H5 hemagglutinin binds with high avidity to sialic acid on different O-linked core structures on mucin-type fusion proteins. 24233973 7 88 gly sialylated 1433:1442 arg1 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 5 89 theme H5 1198:1199 arg1 partner 1187:1193 a good binding partner 1172:1193 a good binding partner of H5 1172:1199 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 5 89 theme H5 1198:1199 arg1 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG(2b) 1153:1167 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 3 90 theme FUT-VII 929:935 arg1 glycosyltransferases 937:956 the core 2 β1,6GnT-I and FUT-VII glycosyltransferases 904:956 the core 2 β1,6GnT-I and FUT-VII glycosyltransferases 904:956 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 2 91 theme fusion 464:469 arg1 protein 471:477 The fusion protein 460:477 The fusion protein 460:477 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 1 92 theme receptor 299:306 arg1 Siaα2-3Gal 308:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 the influenza hemagglutinin receptor Siaα2-3Gal 271:317 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 3 93 theme β1,6GnT-I 915:923 arg1 glycosyltransferases 937:956 the core 2 β1,6GnT-I and FUT-VII glycosyltransferases 904:956 the core 2 β1,6GnT-I and FUT-VII glycosyltransferases 904:956 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 7 94 theme similar 1317:1323 arg1 strength 1325:1332 similar strength 1317:1332 similar strength 1317:1332 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 2 95 theme cell 502:505 arg1 lines 507:511 stable cell lines 495:511 stable cell lines in large scale cultures 495:535 The fusion protein was produced by stable cell lines in large scale cultures and purified with affinity- and gel filtration chromatography. 24233973 3 96 theme hamster 675:681 arg1 CHO 690:692 CHO 690:692 CHO 690:692 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 3 96 theme hamster 675:681 arg1 ovary 683:687 the Chinese hamster ovary 663:687 the Chinese hamster ovary (CHO)-K1 663:696 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 5 97 theme good 1174:1177 arg1 partner 1187:1193 a good binding partner 1172:1193 a good binding partner of H5 1172:1199 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 5 97 theme good 1174:1177 arg1 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG 1153:1163 PSGL-1/mIgG(2b) 1153:1167 Biacore experiments revealed that PSGL-1/mIgG(2b) is a good binding partner of H5. 24233973 1 98 theme O-glycan 332:339 arg1 chains 341:346 different O-glycan chains 322:346 different O-glycan chains 322:346 The interaction between P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL-1/mIgG(2b)) fusion protein carrying multiple copies of the influenza hemagglutinin receptor Siaα2-3Gal on different O-glycan chains and recombinant human influenza H5N1 A/Vietnam/1203/04 hemagglutinin was investigated with a Biacore biosensor. 24233973 7 99 theme clone 1504:1508 arg1 lactosamine 1491:1501 mainly sialylated lactosamine 1473:1501 mainly sialylated lactosamine (clone C-PSLex) 1473:1517 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 99 theme clone 1504:1508 arg1 C-PSLex 1510:1516 clone C-PSLex 1504:1516 clone C-PSLex 1504:1516 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 7 100 theme H5 1289:1290 arg1 hemagglutinin 1292:1304 The H5 hemagglutinin 1285:1304 The H5 hemagglutinin 1285:1304 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24233973 4 101 theme non-derivatized 1092:1106 arg1 O-glycans 1108:1116 released non-derivatized O-glycans 1083:1116 released non-derivatized O-glycans 1083:1116 Glycosylation was characterized by lectin Western blotting of the proteins and liquid chromatography - mass spectrometry of released non-derivatized O-glycans. 24233973 3 102 dep ovary 683:687 arg1 -K1 694:696 -K1 694:696 the Chinese hamster ovary (CHO)-K1 663:696 TheC-P55 and 293-P cell lines were established by transfecting the Chinese hamster ovary (CHO)-K1 and Human embryonic kidney (HEK)-293 cell lines with plasmids encoding the PSGL-1/mIgG(2b) fusion protein, while the C-PSLex cell line was engineered by transfecting CHO-K1 cells with the plasmids encoding the core 2 β1,6GnT-I and FUT-VII glycosyltransferases. 24233973 7 103 theme lactosamine 1491:1501 arg1 O-glycans 1519:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans 1433:1527 The H5 hemagglutinin binds with similar strength to PSGL-1/mIgG(2b) carrying mostly sialylated core 1 (clone C-P55), a mix of sialylated core 1 and sialylated lactosamine (clone 293-P) or mainly sialylated lactosamine (clone C-PSLex) O-glycans, indicating that this hemagglutinin is unable to discriminate between these structures.The potential use of the large, flexible PSGL-1/mIgG(2b) mucin-type fusion protein carrying Siaα2-3Gal as a multivalent inhibitor of influenza virus is discussed. 24385394 5 0 theme synthesis 653:661 arg1 by-product 639:648 a by-product 637:648 a by-product of synthesis 637:661 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 5 0 theme synthesis 653:661 arg1 linker 629:634 a readily released reducing-end linker 597:634 a readily released reducing-end linker 597:634 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 5 1 theme collaborating 527:539 arg1 colleagues 541:550 collaborating colleagues 527:550 collaborating colleagues 527:550 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 1 2 theme accepted 155:162 arg1 protocols 164:172 accepted protocols 155:172 accepted protocols 155:172 Documenting mass spectral data is a fundamental aspect of accepted protocols. 24385394 4 3 theme HexNAc 461:466 arg1 linkage 479:485 a CID-labile HexNAc glycosidic linkage 448:485 a CID-labile HexNAc glycosidic linkage 448:485 The latter are most frequently pendant through a CID-labile HexNAc glycosidic linkage. 24385394 1 4 theme protocols 164:172 arg1 aspect 145:150 a fundamental aspect 131:150 a fundamental aspect of accepted protocols 131:172 Documenting mass spectral data is a fundamental aspect of accepted protocols. 24385394 9 5 theme various 1047:1053 arg1 Lewis 1055:1059 various Lewis and H antigens 1047:1074 Lewis 1055:1059 Included in this study were various Lewis and H antigens, 3- and 6-linked sialyl-lactosamine, NeuAc-2,8-NeuAc dimer, and Galα1,3Gal. 24385394 5 6 theme released 607:614 arg1 by-product 639:648 a by-product 637:648 a by-product of synthesis 637:661 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 5 6 theme released 607:614 arg1 linker 629:634 a readily released reducing-end linker 597:634 a readily released reducing-end linker 597:634 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 2 7 theme glycan 276:281 arg1 epitopes 283:290 natural and synthetic glycan epitopes 254:290 natural and synthetic glycan epitopes 254:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 10 8 theme reproducible 1253:1264 arg1 spectra 1284:1290 reproducible high signal/noise spectra 1253:1290 reproducible high signal/noise spectra for spectral matching 1253:1312 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 3 9 located found 330:334 arg1 N- 371:372 N- 371:372 N- 371:372 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 9 located found 330:334 arg2 epitopes 297:304 The epitopes 293:304 The epitopes considered 293:315 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 9 located found 330:334 arg1 termini 349:355 conjugate termini 339:355 conjugate termini of lipids 339:365 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 9 located found 330:334 arg2 clusters 321:328 clusters 321:328 clusters found on conjugate termini of lipids and N- and O-glycans of proteins 321:398 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 5 10 theme reducing-end 616:627 arg1 by-product 639:648 a by-product 637:648 a by-product of synthesis 637:661 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 5 10 theme reducing-end 616:627 arg1 linker 629:634 a readily released reducing-end linker 597:634 a readily released reducing-end linker 597:634 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 2 11 theme synthetic 266:274 arg1 epitopes 283:290 natural and synthetic glycan epitopes 254:290 natural and synthetic glycan epitopes 254:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 10 12 theme spectral 1190:1197 arg1 data 1199:1202 high quality spectral data 1177:1202 high quality spectral data 1177:1202 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 4 13 theme CID-labile 450:459 arg1 linkage 479:485 a CID-labile HexNAc glycosidic linkage 448:485 a CID-labile HexNAc glycosidic linkage 448:485 The latter are most frequently pendant through a CID-labile HexNAc glycosidic linkage. 24385394 6 14 dep methylated 692:701 arg1 extracted 704:712 extracted 704:712 extracted 704:712 All samples were comparably methylated, extracted, and MS(n) disassembled to compare their linkage and branching spectral details. 24385394 12 15 theme mass 1447:1450 arg1 spectrometers 1452:1464 several mass spectrometers 1439:1464 several mass spectrometers 1439:1464 This study includes comparability statistics and evaluations on several mass spectrometers. 24385394 7 16 theme disassembly 853:863 arg1 pathway 865:871 a disassembly pathway 851:871 a disassembly pathway 851:871 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 2 17 from contrast 194:201 arg1 report 183:188 this report 178:188 this report 178:188 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 8 18 theme linkage 996:1002 arg1 details 985:991 the details 981:991 the details of linkage and branching 981:1016 Further steps of disassembly are necessary to confirm the details of linkage and branching. 24385394 2 19 theme natural 254:260 arg1 epitopes 283:290 natural and synthetic glycan epitopes 254:290 natural and synthetic glycan epitopes 254:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 5 20 theme synthetic 492:500 arg1 samples 502:508 The synthetic samples 488:508 The synthetic samples 488:508 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 11 21 theme adducted 1351:1358 arg1 samples 1319:1325 All samples 1315:1325 All samples 1315:1325 All samples were analyzed as sodium adducted positive ions. 24385394 11 21 theme adducted 1351:1358 arg1 ions 1369:1372 sodium adducted positive ions 1344:1372 sodium adducted positive ions 1344:1372 All samples were analyzed as sodium adducted positive ions. 24385394 4 22 theme glycosidic 468:477 arg1 linkage 479:485 a CID-labile HexNAc glycosidic linkage 448:485 a CID-labile HexNAc glycosidic linkage 448:485 The latter are most frequently pendant through a CID-labile HexNAc glycosidic linkage. 24385394 11 23 theme positive 1360:1367 arg1 samples 1319:1325 All samples 1315:1325 All samples 1315:1325 All samples were analyzed as sodium adducted positive ions. 24385394 11 23 theme positive 1360:1367 arg1 ions 1369:1372 sodium adducted positive ions 1344:1372 sodium adducted positive ions 1344:1372 All samples were analyzed as sodium adducted positive ions. 24385394 6 24 theme spectral 777:784 arg1 details 786:792 their linkage and branching spectral details 749:792 details 786:792 All samples were comparably methylated, extracted, and MS(n) disassembled to compare their linkage and branching spectral details. 24385394 12 25 theme comparability 1395:1407 arg1 statistics 1409:1418 comparability statistics 1395:1418 comparability statistics 1395:1418 This study includes comparability statistics and evaluations on several mass spectrometers. 24385394 12 26 from evaluations 1424:1434 arg1 spectrometers 1452:1464 several mass spectrometers 1439:1464 several mass spectrometers 1439:1464 This study includes comparability statistics and evaluations on several mass spectrometers. 24385394 0 27 theme Structural 0:9 arg1 documentation 11:23 Structural documentation 0:23 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 0:95 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 7 28 theme sample 800:805 arg1 types 807:811 Both sample types 795:811 Both sample types 795:811 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 6 29 theme branching 767:775 arg1 details 786:792 their linkage and branching spectral details 749:792 details 786:792 All samples were comparably methylated, extracted, and MS(n) disassembled to compare their linkage and branching spectral details. 24385394 7 30 theme type 827:830 arg1 fragments 832:840 B-ion type fragments 821:840 B-ion type fragments early in a disassembly pathway 821:871 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 8 31 theme branching 1008:1016 arg1 details 985:991 the details 981:991 the details of linkage and branching 981:1016 Further steps of disassembly are necessary to confirm the details of linkage and branching. 24385394 0 32 theme glycan 28:33 arg1 epitopes 35:42 glycan epitopes 28:42 glycan epitopes 28:42 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 7 33 theme structure 916:924 arg1 compositions 883:894 their compositions 877:894 their compositions 877:894 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 7 33 theme structure 916:924 arg1 suggestion 902:911 a suggestion 900:911 a suggestion of structure 900:924 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 1 34 theme mass 109:112 arg1 data 123:126 mass spectral data 109:126 mass spectral data 109:126 Documenting mass spectral data is a fundamental aspect of accepted protocols. 24385394 12 35 from statistics 1409:1418 arg1 spectrometers 1452:1464 several mass spectrometers 1439:1464 several mass spectrometers 1439:1464 This study includes comparability statistics and evaluations on several mass spectrometers. 24385394 11 36 theme sodium 1344:1349 arg1 samples 1319:1325 All samples 1315:1325 All samples 1315:1325 All samples were analyzed as sodium adducted positive ions. 24385394 11 36 theme sodium 1344:1349 arg1 ions 1369:1372 sodium adducted positive ions 1344:1372 sodium adducted positive ions 1344:1372 All samples were analyzed as sodium adducted positive ions. 24385394 1 37 theme spectral 114:121 arg1 data 123:126 mass spectral data 109:126 mass spectral data 109:126 Documenting mass spectral data is a fundamental aspect of accepted protocols. 24385394 9 38 theme 6-linked 1084:1091 arg1 sialyl-lactosamine 1093:1110 6-linked sialyl-lactosamine 1084:1110 6-linked sialyl-lactosamine 1084:1110 Included in this study were various Lewis and H antigens, 3- and 6-linked sialyl-lactosamine, NeuAc-2,8-NeuAc dimer, and Galα1,3Gal. 24385394 3 39 theme proteins 391:398 arg1 N- 371:372 N- 371:372 N- 371:372 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 39 theme proteins 391:398 arg1 termini 349:355 conjugate termini 339:355 conjugate termini of lipids 339:365 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 39 theme proteins 391:398 arg1 O-glycans 378:386 O-glycans 378:386 O-glycans 378:386 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 9 40 theme H 1065:1065 arg1 antigens 1067:1074 various Lewis and H antigens 1047:1074 antigens 1067:1074 Included in this study were various Lewis and H antigens, 3- and 6-linked sialyl-lactosamine, NeuAc-2,8-NeuAc dimer, and Galα1,3Gal. 24385394 10 41 theme high 1266:1269 arg1 spectra 1284:1290 reproducible high signal/noise spectra 1253:1290 reproducible high signal/noise spectra for spectral matching 1253:1312 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 2 42 theme sequential 209:218 arg1 spectra 232:238 MS(n) sequential disassembly spectra 203:238 MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes 203:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 7 43 theme B-ion 821:825 arg1 fragments 832:840 B-ion type fragments 821:840 B-ion type fragments early in a disassembly pathway 821:871 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 0 44 theme epitopes 35:42 arg1 documentation 11:23 Structural documentation 0:23 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 0:95 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 10 45 theme Sample 1152:1157 arg1 infusion 1159:1166 Sample infusion 1152:1166 Sample infusion 1152:1166 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 10 46 theme signal/noise 1271:1282 arg1 spectra 1284:1290 reproducible high signal/noise spectra 1253:1290 reproducible high signal/noise spectra for spectral matching 1253:1312 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 3 47 theme lipids 360:365 arg1 N- 371:372 N- 371:372 N- 371:372 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 47 theme lipids 360:365 arg1 termini 349:355 conjugate termini 339:355 conjugate termini of lipids 339:365 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 47 theme lipids 360:365 arg1 O-glycans 378:386 O-glycans 378:386 O-glycans 378:386 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 3 48 theme conjugate 339:347 arg1 termini 349:355 conjugate termini 339:355 conjugate termini of lipids 339:365 The epitopes considered are clusters found on conjugate termini of lipids and N- and O-glycans of proteins. 24385394 0 49 theme mass 56:59 arg1 spectrometry 61:72 sequential mass spectrometry 45:72 sequential mass spectrometry 45:72 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 2 50 theme n 206:206 arg1 spectra 232:238 MS(n) sequential disassembly spectra 203:238 MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes 203:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 9 51 theme NeuAc-2,8-NeuAc 1113:1127 arg1 dimer 1129:1133 NeuAc-2,8-NeuAc dimer 1113:1133 NeuAc-2,8-NeuAc dimer 1113:1133 Included in this study were various Lewis and H antigens, 3- and 6-linked sialyl-lactosamine, NeuAc-2,8-NeuAc dimer, and Galα1,3Gal. 24385394 0 52 theme sequential 45:54 arg1 spectrometry 61:72 sequential mass spectrometry 45:72 sequential mass spectrometry 45:72 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 2 53 theme MS 203:204 arg1 spectra 232:238 MS(n) sequential disassembly spectra 203:238 MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes 203:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24385394 5 54 theme commercial 556:565 arg1 sources 567:573 commercial sources 556:573 commercial sources 556:573 The synthetic samples were supplied by collaborating colleagues and commercial sources and usually possessed a readily released reducing-end linker, a by-product of synthesis. 24385394 7 55 from early 842:846 arg1 pathway 865:871 a disassembly pathway 851:871 a disassembly pathway 851:871 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 8 56 theme Further 927:933 arg1 steps 935:939 Further steps 927:939 Further steps of disassembly 927:954 Further steps of disassembly are necessary to confirm the details of linkage and branching. 24385394 10 57 theme high 1177:1180 arg1 data 1199:1202 high quality spectral data 1177:1202 high quality spectral data 1177:1202 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 10 58 theme quality 1182:1188 arg1 data 1199:1202 high quality spectral data 1177:1202 high quality spectral data 1177:1202 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 12 59 theme several 1439:1445 arg1 spectrometers 1452:1464 several mass spectrometers 1439:1464 several mass spectrometers 1439:1464 This study includes comparability statistics and evaluations on several mass spectrometers. 24385394 7 60 from pathway 865:871 arg1 early 842:846 early 842:846 early 842:846 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 8 61 theme disassembly 944:954 arg1 steps 935:939 Further steps 927:939 Further steps of disassembly 927:954 Further steps of disassembly are necessary to confirm the details of linkage and branching. 24385394 1 62 theme fundamental 133:143 arg1 aspect 145:150 a fundamental aspect 131:150 a fundamental aspect of accepted protocols 131:172 Documenting mass spectral data is a fundamental aspect of accepted protocols. 24385394 10 63 theme small 1227:1231 arg1 fragments 1233:1241 small fragments 1227:1241 small fragments 1227:1241 Sample infusion provided high quality spectral data whereas disassembly to small fragments generates reproducible high signal/noise spectra for spectral matching. 24385394 7 64 theme early 842:846 arg1 fragments 832:840 B-ion type fragments 821:840 B-ion type fragments early in a disassembly pathway 821:871 Both sample types provide B-ion type fragments early in a disassembly pathway and their compositions are a suggestion of structure. 24385394 0 65 dep documentation 11:23 arg1 spectral 78:85 spectral 78:85 spectral 78:85 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 0 65 dep documentation 11:23 arg1 spectrometry 61:72 sequential mass spectrometry 45:72 sequential mass spectrometry 45:72 Structural documentation of glycan epitopes: sequential mass spectrometry and spectral matching. 24385394 9 66 link 6-linked 1084:1091 arg1 sialyl-lactosamine 1093:1110 6-linked sialyl-lactosamine 1084:1110 6-linked sialyl-lactosamine 1084:1110 Included in this study were various Lewis and H antigens, 3- and 6-linked sialyl-lactosamine, NeuAc-2,8-NeuAc dimer, and Galα1,3Gal. 24385394 2 67 theme disassembly 220:230 arg1 spectra 232:238 MS(n) sequential disassembly spectra 203:238 MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes 203:290 In this report, we contrast MS(n) sequential disassembly spectra obtained from natural and synthetic glycan epitopes. 24680511 9 0 theme relative 1666:1673 arg1 distribution 1675:1686 the relative distribution 1662:1686 the relative distribution of CPS and LOS structures 1662:1712 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 12 1 theme host 2052:2055 arg1 tropism 2062:2068 host cell tropism 2052:2068 the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures 2002:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 2 2 from humans 366:371 arg1 pathogenic 352:361 pathogenic 352:361 pathogenic 352:361 While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. 24680511 10 3 theme mass 1776:1779 arg1 species 1781:1787 LOS molecular mass species 1762:1787 LOS molecular mass species distribution between temperatures 1762:1821 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 11 4 theme plate 1933:1937 arg1 assay 1953:1957 the plate agglutination assay 1929:1957 the plate agglutination assay 1929:1957 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 10 5 theme LOS 1762:1764 arg1 species 1781:1787 LOS molecular mass species 1762:1787 LOS molecular mass species distribution between temperatures 1762:1821 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 9 6 theme structures 1703:1712 arg1 distribution 1675:1686 the relative distribution 1662:1686 the relative distribution of CPS and LOS structures 1662:1712 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 12 7 theme C. 2042:2043 arg1 jejuni 2045:2050 C. jejuni 2042:2050 C. jejuni 2042:2050 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 6 8 theme avian 982:986 arg1 conditions 993:1002 human and avian host conditions 972:1002 human and avian host conditions 972:1002 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 10 9 from change 1752:1757 arg1 distribution 1789:1800 LOS molecular mass species distribution 1762:1800 LOS molecular mass species distribution between temperatures 1762:1821 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 4 10 theme important 670:678 arg1 stimulus 680:687 an important stimulus 667:687 an important stimulus for specific adaptation 667:711 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 7 11 dep jejuni 1136:1141 arg1 profiles 1149:1156 81116 profiles 1143:1156 C. jejuni 81116 profiles 1133:1156 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 6 12 theme human 972:976 arg1 conditions 993:1002 human and avian host conditions 972:1002 human and avian host conditions 972:1002 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 5 13 theme total 757:761 arg1 glycome 771:777 the total surface glycome 753:777 the total surface glycome of C. jejuni in response to temperature 753:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 10 14 theme SDS-PAGE 1840:1847 arg1 analysis 1849:1856 SDS-PAGE analysis 1840:1856 SDS-PAGE analysis 1840:1856 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 1 15 theme infectious 260:269 arg1 trigger 271:277 the most frequently identified infectious trigger 229:277 the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 229:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 15 theme infectious 260:269 arg1 humans 208:213 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 12 16 theme important 2006:2014 arg1 glycosylation 2016:2028 the important glycosylation 2002:2028 the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures 2002:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 8 17 theme strains 1480:1486 arg1 distinguishable 1504:1518 distinguishable 1504:1518 distinguishable 1504:1518 The profiles of both strains varied and were distinguishable at both temperatures. 24680511 8 17 theme strains 1480:1486 arg1 profiles 1463:1470 The profiles 1459:1470 The profiles of both strains 1459:1486 The profiles of both strains varied and were distinguishable at both temperatures. 24680511 12 18 theme microarray 2132:2141 arg1 platform 2143:2150 the lectin microarray platform 2121:2150 the lectin microarray platform 2121:2150 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 5 19 from response 795:802 arg1 modulation 739:748 the modulation 735:748 the modulation of the total surface glycome of C. jejuni in response to temperature 735:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 10 20 theme species 1781:1787 arg1 distribution 1789:1800 LOS molecular mass species distribution 1762:1800 LOS molecular mass species distribution between temperatures 1762:1821 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 9 21 theme higher 1549:1554 arg1 temperature 1556:1566 the higher temperature 1545:1566 the higher temperature 1545:1566 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 7 22 from 37°C 1453:1456 arg1 recognised 1421:1430 recognised 1421:1430 recognised 1421:1430 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 0 23 theme lectin 101:106 arg1 microarray 108:117 lectin microarray 101:117 lectin microarray 101:117 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 5 24 theme glycome 771:777 arg1 modulation 739:748 the modulation 735:748 the modulation of the total surface glycome of C. jejuni in response to temperature 735:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 5 25 theme C. 782:783 arg1 jejuni 785:790 C. jejuni 782:790 C. jejuni 782:790 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 3 26 theme Bacterial 408:416 arg1 carbohydrates 431:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 26 theme Bacterial 408:416 arg1 factors 469:475 important virulence factors 449:475 important virulence factors 449:475 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 5 27 theme jejuni 785:790 arg1 glycome 771:777 the total surface glycome 753:777 the total surface glycome of C. jejuni in response to temperature 753:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 3 28 theme virulence 459:467 arg1 carbohydrates 431:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 28 theme virulence 459:467 arg1 factors 469:475 important virulence factors 449:475 important virulence factors 449:475 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 0 29 theme Campylobacter 0:12 arg1 discrimination 28:41 strain discrimination 21:41 strain discrimination 21:41 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 29 theme Campylobacter 0:12 arg1 jejuni 14:19 Campylobacter jejuni 0:19 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression 0:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 29 theme Campylobacter 0:12 arg1 expression 77:86 temperature-dependent glycome expression 47:86 temperature-dependent glycome expression 47:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 1 30 theme Gram-negative 120:132 arg1 cause 170:174 the leading cause 158:174 the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 158:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 30 theme Gram-negative 120:132 arg1 jejuni 148:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 0 31 theme strain 21:26 arg1 discrimination 28:41 strain discrimination 21:41 strain discrimination 21:41 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 31 theme strain 21:26 arg1 jejuni 14:19 Campylobacter jejuni 0:19 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression 0:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 6 32 theme jejuni 899:904 arg1 strains 906:912 C. jejuni strains 81116 and 81-176 896:929 strains 906:912 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 6 33 theme lectin 1052:1057 arg1 microarrays 1059:1069 lectin microarrays 1052:1069 lectin microarrays constructed to include a wide range of binding specificities 1052:1130 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 4 34 theme jejuni 603:608 arg1 infection 560:568 infection 560:568 infection 560:568 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 4 34 theme jejuni 603:608 arg1 colonisation 584:595 persistent colonisation 573:595 persistent colonisation 573:595 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 3 35 theme surface 423:429 arg1 carbohydrates 431:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 35 theme surface 423:429 arg1 factors 469:475 important virulence factors 449:475 important virulence factors 449:475 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 1 36 dep humans 208:213 arg1 trigger 271:277 the most frequently identified infectious trigger 229:277 the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 229:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 36 dep humans 208:213 arg1 worldwide 215:223 worldwide 215:223 worldwide 215:223 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 36 dep humans 208:213 arg1 humans 208:213 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 11 37 theme certain 1895:1901 arg1 lectins 1903:1909 certain lectins 1895:1909 certain lectins 1895:1909 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 0 38 theme glycome 69:75 arg1 jejuni 14:19 Campylobacter jejuni 0:19 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression 0:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 38 theme glycome 69:75 arg1 expression 77:86 temperature-dependent glycome expression 47:86 temperature-dependent glycome expression 47:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 9 39 theme structure 1603:1611 arg1 dominance 1577:1585 reduced dominance 1569:1585 reduced dominance of the LPS-like structure 1569:1611 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 4 40 theme persistent 573:582 arg1 colonisation 584:595 persistent colonisation 573:595 persistent colonisation 573:595 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 7 41 theme surface 1301:1307 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 41 theme surface 1301:1307 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 41 theme surface 1301:1307 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 4 42 theme host 638:641 arg1 temperature 643:653 host temperature 638:653 host temperature 638:653 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 2 43 theme avian 395:399 arg1 hosts 401:405 avian hosts 395:405 avian hosts 395:405 While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. 24680511 12 44 from temperatures 2090:2101 arg1 tropism 2062:2068 host cell tropism 2052:2068 the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures 2002:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 7 45 theme recognised 1285:1294 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 45 theme recognised 1285:1294 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 45 theme recognised 1285:1294 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 46 theme C. 1133:1134 arg1 jejuni 1136:1141 C. jejuni 1133:1141 C. jejuni 81116 profiles 1133:1156 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 1 47 theme Guillain-Barré 302:315 arg1 syndrome 317:324 Guillain-Barré syndrome 302:324 Guillain-Barré syndrome (GBS) 302:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 47 theme Guillain-Barré 302:315 arg1 GBS 327:329 GBS 327:329 GBS 327:329 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 12 48 theme growth 2083:2088 arg1 temperatures 2090:2101 different growth temperatures 2073:2101 different growth temperatures 2073:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 6 49 theme wide 1096:1099 arg1 range 1101:1105 a wide range 1094:1105 a wide range of binding specificities 1094:1130 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 2 50 theme C. 339:340 arg1 jejuni 342:347 C. jejuni 339:347 C. jejuni 339:347 While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. 24680511 5 51 from modulation 739:748 arg1 response 795:802 response 795:802 response to temperature 795:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 9 52 theme CPS 1691:1693 arg1 distribution 1675:1686 the relative distribution 1662:1686 the relative distribution of CPS and LOS structures 1662:1712 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 10 53 theme molecular 1766:1774 arg1 species 1781:1787 LOS molecular mass species 1762:1787 LOS molecular mass species distribution between temperatures 1762:1821 This change in LOS molecular mass species distribution between temperatures was confirmed by SDS-PAGE analysis. 24680511 12 54 theme cell 2057:2060 arg1 tropism 2062:2068 host cell tropism 2052:2068 the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures 2002:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 9 55 theme LOS 1699:1701 arg1 structures 1703:1712 LOS structures 1699:1712 LOS structures 1699:1712 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 2 56 from commensal 382:390 arg1 hosts 401:405 avian hosts 395:405 avian hosts 395:405 While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. 24680511 4 57 theme specific 693:700 arg1 adaptation 702:711 specific adaptation 693:711 specific adaptation 693:711 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 6 58 theme host 988:991 arg1 conditions 993:1002 human and avian host conditions 972:1002 human and avian host conditions 972:1002 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 5 59 theme modulation 739:748 arg1 examination 720:730 examination 720:730 examination of the modulation of the total surface glycome of C. jejuni in response to temperature 720:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 7 60 link N-linked 1245:1252 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 60 link N-linked 1245:1252 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 60 link N-linked 1245:1252 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 5 61 theme pathogenic 856:865 arg1 mechanisms 867:876 commensal and pathogenic mechanisms 842:876 commensal and pathogenic mechanisms for this species 842:893 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 1 62 theme identified 249:258 arg1 trigger 271:277 the most frequently identified infectious trigger 229:277 the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 229:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 62 theme identified 249:258 arg1 humans 208:213 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 11 63 theme opposite 1873:1880 arg1 behaviour 1882:1890 opposite behaviour 1873:1890 opposite behaviour of certain lectins 1873:1909 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 5 64 from glycome 771:777 arg1 response 795:802 response 795:802 response to temperature 795:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 12 65 theme lectin 2125:2130 arg1 platform 2143:2150 the lectin microarray platform 2121:2150 the lectin microarray platform 2121:2150 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 1 66 from trigger 271:277 arg1 patients 282:289 patients 282:289 patients developing Guillain-Barré syndrome (GBS) 282:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 67 theme leading 162:168 arg1 cause 170:174 the leading cause 158:174 the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 158:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 67 theme leading 162:168 arg1 jejuni 148:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 7 68 theme N-linked 1388:1395 arg1 glycosylation 1397:1409 N-linked glycosylation 1388:1409 N-linked glycosylation 1388:1409 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 69 theme N-linked 1245:1252 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 69 theme N-linked 1245:1252 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 69 theme N-linked 1245:1252 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 8 70 from temperatures 1528:1539 arg1 distinguishable 1504:1518 distinguishable 1504:1518 distinguishable 1504:1518 The profiles of both strains varied and were distinguishable at both temperatures. 24680511 8 70 from temperatures 1528:1539 arg1 profiles 1463:1470 The profiles 1459:1470 The profiles of both strains 1459:1486 The profiles of both strains varied and were distinguishable at both temperatures. 24680511 6 71 theme whole 1023:1027 arg1 cells 1029:1033 whole cells 1023:1033 whole cells 1023:1033 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 7 72 theme -like 1226:1230 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 72 theme -like 1226:1230 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 72 theme -like 1226:1230 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 1 73 from worldwide 215:223 arg1 patients 282:289 patients 282:289 patients developing Guillain-Barré syndrome (GBS) 282:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 74 theme gastroenteritis 189:203 arg1 cause 170:174 the leading cause 158:174 the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 158:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 74 theme gastroenteritis 189:203 arg1 jejuni 148:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 5 75 theme surface 763:769 arg1 glycome 771:777 the total surface glycome 753:777 the total surface glycome of C. jejuni in response to temperature 753:817 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 9 76 theme reduced 1569:1575 arg1 dominance 1577:1585 reduced dominance 1569:1585 reduced dominance of the LPS-like structure 1569:1611 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 2 77 from pathogenic 352:361 arg1 humans 366:371 humans 366:371 humans 366:371 While C. jejuni is pathogenic in humans, it is a commensal in avian hosts. 24680511 6 78 theme specificities 1118:1130 arg1 range 1101:1105 a wide range 1094:1105 a wide range of binding specificities 1094:1130 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 7 79 theme characterised 1188:1200 arg1 LPS 1222:1224 LPS 1222:1224 LPS 1222:1224 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 79 theme characterised 1188:1200 arg1 lipopolysaccharide 1202:1219 the previously characterised lipopolysaccharide 1173:1219 the previously characterised lipopolysaccharide (LPS) 1173:1225 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 1 80 from cause 170:174 arg1 humans 208:213 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 208:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 80 from cause 170:174 arg1 trigger 271:277 the most frequently identified infectious trigger 229:277 the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 229:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 80 from cause 170:174 arg1 worldwide 215:223 worldwide 215:223 worldwide 215:223 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 9 81 theme LPS-like 1594:1601 arg1 structure 1603:1611 the LPS-like structure 1590:1611 the LPS-like structure 1590:1611 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 9 82 from change 1652:1657 arg1 distribution 1675:1686 the relative distribution 1662:1686 the relative distribution of CPS and LOS structures 1662:1712 At the higher temperature, reduced dominance of the LPS-like structure was associated with strain 81116 and a change in the relative distribution of CPS and LOS structures was indicated for strain 81-176. 24680511 3 83 theme cell 418:421 arg1 carbohydrates 431:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 83 theme cell 418:421 arg1 factors 469:475 important virulence factors 449:475 important virulence factors 449:475 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 4 84 theme C. 600:601 arg1 jejuni 603:608 C. jejuni 600:608 C. jejuni 600:608 The mechanisms leading to infection or persistent colonisation of C. jejuni are not well understood but host temperature may provide an important stimulus for specific adaptation. 24680511 12 85 dep glycosylation 2016:2028 arg1 tropism 2062:2068 host cell tropism 2052:2068 the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures 2002:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 11 86 theme agglutination 1939:1951 arg1 assay 1953:1957 the plate agglutination assay 1929:1957 the plate agglutination assay 1929:1957 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 5 87 theme commensal 842:850 arg1 mechanisms 867:876 commensal and pathogenic mechanisms 842:876 commensal and pathogenic mechanisms for this species 842:893 Thus, examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species. 24680511 1 88 theme Campylobacter 134:146 arg1 cause 170:174 the leading cause 158:174 the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS) 158:330 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 1 88 theme Campylobacter 134:146 arg1 jejuni 148:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni 120:153 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 7 89 link N-linked 1388:1395 arg1 glycosylation 1397:1409 N-linked glycosylation 1388:1409 N-linked glycosylation 1388:1409 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 0 90 theme temperature-dependent 47:67 arg1 jejuni 14:19 Campylobacter jejuni 0:19 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression 0:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 90 theme temperature-dependent 47:67 arg1 expression 77:86 temperature-dependent glycome expression 47:86 temperature-dependent glycome expression 47:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 6 91 theme C. 896:897 arg1 strains 906:912 C. jejuni strains 81116 and 81-176 896:929 strains 906:912 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 24680511 7 92 theme capsular 1326:1333 arg1 CPS 1351:1353 CPS 1351:1353 CPS 1351:1353 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 92 theme capsular 1326:1333 arg1 polysaccharide 1335:1348 capsular polysaccharide 1326:1348 capsular polysaccharide (CPS) 1326:1354 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 3 93 theme important 449:457 arg1 carbohydrates 431:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates 408:443 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 93 theme important 449:457 arg1 factors 469:475 important virulence factors 449:475 important virulence factors 449:475 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 94 from roles 486:490 arg1 infection 523:531 infection 523:531 infection 523:531 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 94 from roles 486:490 arg1 colonisation 506:517 colonisation 506:517 colonisation 506:517 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 3 94 from roles 486:490 arg1 adherence 495:503 adherence 495:503 adherence 495:503 Bacterial cell surface carbohydrates are important virulence factors and play roles in adherence, colonisation and infection. 24680511 11 95 theme microarray 1967:1976 arg1 platform 1978:1985 the microarray platform 1963:1985 the microarray platform 1963:1985 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 1 96 theme bacterial 179:187 arg1 gastroenteritis 189:203 bacterial gastroenteritis 179:203 bacterial gastroenteritis 179:203 Gram-negative Campylobacter jejuni is the leading cause of bacterial gastroenteritis in humans worldwide and the most frequently identified infectious trigger in patients developing Guillain-Barré syndrome (GBS). 24680511 11 97 theme lectins 1903:1909 arg1 behaviour 1882:1890 opposite behaviour 1873:1890 opposite behaviour of certain lectins 1873:1909 Additionally, opposite behaviour of certain lectins was noted between the plate agglutination assay and the microarray platform. 24680511 7 98 theme cell 1296:1299 arg1 structures 1309:1318 the predominantly recognised cell surface structures 1267:1318 the predominantly recognised cell surface structures 1267:1318 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 98 theme cell 1296:1299 arg1 molecule 1232:1239 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 molecule 1232:1239 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 7 98 theme cell 1296:1299 arg1 glycans 1254:1260 the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans 1173:1260 glycans 1254:1260 C. jejuni 81116 profiles indicated that the previously characterised lipopolysaccharide (LPS)-like molecule and N-linked glycans were the predominantly recognised cell surface structures while capsular polysaccharide (CPS), lipooligosaccharides (LOS) and N-linked glycosylation were best recognised for strain 81-176 at 37°C. 24680511 12 99 theme different 2073:2081 arg1 temperatures 2090:2101 different growth temperatures 2073:2101 different growth temperatures 2073:2101 Insights into the important glycosylation involved in C. jejuni host cell tropism at different growth temperatures were gained using the lectin microarray platform. 24680511 0 100 dep jejuni 14:19 arg1 discrimination 28:41 strain discrimination 21:41 strain discrimination 21:41 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 100 dep jejuni 14:19 arg1 jejuni 14:19 Campylobacter jejuni 0:19 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression 0:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 0 100 dep jejuni 14:19 arg1 expression 77:86 temperature-dependent glycome expression 47:86 temperature-dependent glycome expression 47:86 Campylobacter jejuni strain discrimination and temperature-dependent glycome expression profiling by lectin microarray. 24680511 6 101 theme binding 1110:1116 arg1 specificities 1118:1130 binding specificities 1110:1130 binding specificities 1110:1130 C. jejuni strains 81116 and 81-176 were cultured at 37 and 42°C to simulate human and avian host conditions, respectively, and whole cells were profiled on lectin microarrays constructed to include a wide range of binding specificities. 26929048 0 0 theme Amide-Protection 86:101 arg1 Strategy 103:110 an Amide-Protection Strategy 83:110 an Amide-Protection Strategy 83:110 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 1 1 theme Gal 157:159 arg1 [Neu5Ac 166:172 Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans 145:238 [Neu5Ac 166:172 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 1 1 theme Gal 157:159 arg1 tetrasaccharide 128:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 0 2 from Synthesis 10:18 arg1 N-Glycans 65:73 N-Glycans 65:73 N-Glycans 65:73 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 5 3 from Protection 687:696 arg1 residues 737:744 the sialic acid residues 721:744 the sialic acid residues 721:744 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 4 4 gly sialylated 614:623 arg1 disaccharides 625:637 two sialylated disaccharides 610:637 two sialylated disaccharides 610:637 A more-convergent route based on the glycosylation of two sialylated disaccharides was also established to scale up the synthesis. 26929048 3 5 theme synthetic 474:482 arg1 route 484:488 This synthetic route 469:488 This synthetic route 469:488 This synthetic route enabled the synthesis of the as-described disialylated structure. 26929048 5 6 theme sialic 911:916 arg1 residues 923:930 the sialic acid residues 907:930 the sialic acid residues 907:930 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 2 7 theme glucosaminyl 449:460 arg1 donor 462:466 a glucosaminyl donor 447:466 a glucosaminyl donor 447:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 5 8 gly sialylated 826:835 arg1 disaccharides 837:849 the two sialylated disaccharides 818:849 the two sialylated disaccharides 818:849 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 2 9 with disaccharide 429:440 arg1 donor 462:466 a glucosaminyl donor 447:466 a glucosaminyl donor 447:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 1 10 gly disialylated 115:126 arg1 [Neu5Ac 166:172 Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans 145:238 [Neu5Ac 166:172 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 1 10 gly disialylated 115:126 arg1 tetrasaccharide 128:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 5 11 theme glycosylation 787:799 arg1 reaction 801:808 the glycosylation reaction 783:808 the glycosylation reaction 783:808 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 12 theme sialylated 826:835 arg1 disaccharides 837:849 the two sialylated disaccharides 818:849 the two sialylated disaccharides 818:849 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 13 theme sialic 725:730 arg1 residues 737:744 the sialic acid residues 721:744 the sialic acid residues 721:744 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 14 theme bonds 898:902 arg1 presence 877:884 the presence 873:884 the presence of hydrogen bonds on the sialic acid residues 873:930 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 15 theme amide 705:709 arg1 groups 711:716 the amide groups 701:716 the amide groups 701:716 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 2 16 gly sialylated 418:427 arg1 disaccharide 429:440 a sialylated disaccharide 416:440 a sialylated disaccharide with a glucosaminyl donor 416:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 4 17 theme disaccharides 625:637 arg1 glycosylation 593:605 the glycosylation 589:605 the glycosylation of two sialylated disaccharides 589:637 A more-convergent route based on the glycosylation of two sialylated disaccharides was also established to scale up the synthesis. 26929048 0 18 theme Efficient 0:8 arg1 Synthesis 10:18 Efficient Synthesis 0:18 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 0:111 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 4 19 gly glycosylation 593:605 arg1 disaccharides 625:637 two sialylated disaccharides 610:637 two sialylated disaccharides 610:637 A more-convergent route based on the glycosylation of two sialylated disaccharides was also established to scale up the synthesis. 26929048 2 20 theme α-sialylation 298:310 arg1 reaction 312:319 an α-sialylation reaction 295:319 an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor 295:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 5 21 from presence 877:884 arg1 residues 923:930 the sialic acid residues 907:930 the sialic acid residues 907:930 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 3 22 theme disialylated 532:543 arg1 structure 545:553 the as-described disialylated structure 515:553 the as-described disialylated structure 515:553 This synthetic route enabled the synthesis of the as-described disialylated structure. 26929048 1 23 theme disialylated 115:126 arg1 [Neu5Ac 166:172 Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans 145:238 [Neu5Ac 166:172 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 1 23 theme disialylated 115:126 arg1 tetrasaccharide 128:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide 113:142 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 3 24 gly disialylated 532:543 arg1 structure 545:553 the as-described disialylated structure 515:553 the as-described disialylated structure 515:553 This synthetic route enabled the synthesis of the as-described disialylated structure. 26929048 0 25 theme Tetrasaccharide 40:54 arg1 Motif 56:60 the Disialylated Tetrasaccharide Motif 23:60 the Disialylated Tetrasaccharide Motif 23:60 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 5 26 theme reaction 801:808 arg1 yield 774:778 the yield 770:778 the yield of the glycosylation reaction between the two sialylated disaccharides 770:849 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 1 27 dep [Neu5Ac 166:172 arg1 α2,6 174:177 α2,6 174:177 α2,6 174:177 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 1 27 dep [Neu5Ac 166:172 arg1 GlcNAc 180:185 GlcNAc (1) 180:189 GlcNAc (1) 180:189 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 4 28 theme sialylated 614:623 arg1 disaccharides 625:637 two sialylated disaccharides 610:637 two sialylated disaccharides 610:637 A more-convergent route based on the glycosylation of two sialylated disaccharides was also established to scale up the synthesis. 26929048 2 29 theme acid 338:341 arg1 donor 343:347 a sialic acid donor 329:347 a sialic acid donor 329:347 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 0 30 theme Disialylated 27:38 arg1 Motif 56:60 the Disialylated Tetrasaccharide Motif 23:60 the Disialylated Tetrasaccharide Motif 23:60 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 1 31 theme N-glycans 230:238 arg1 termini 214:220 the termini 210:220 the termini of some N-glycans 210:238 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 5 32 theme acid 732:735 arg1 residues 737:744 the sialic acid residues 721:744 the sialic acid residues 721:744 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 2 33 gly glycosylation 399:411 arg1 disaccharide 429:440 a sialylated disaccharide 416:440 a sialylated disaccharide with a glucosaminyl donor 416:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 2 34 theme sialic 331:336 arg1 donor 343:347 a sialic acid donor 329:347 a sialic acid donor 329:347 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 5 35 theme groups 711:716 arg1 Protection 687:696 Protection 687:696 Protection of the amide groups in the sialic acid residues 687:744 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 0 36 theme Motif 56:60 arg1 Synthesis 10:18 Efficient Synthesis 0:18 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 0:111 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 3 37 theme as-described 519:530 arg1 structure 545:553 the as-described disialylated structure 515:553 the as-described disialylated structure 515:553 This synthetic route enabled the synthesis of the as-described disialylated structure. 26929048 0 38 gly Disialylated 27:38 arg1 Motif 56:60 the Disialylated Tetrasaccharide Motif 23:60 the Disialylated Tetrasaccharide Motif 23:60 Efficient Synthesis of the Disialylated Tetrasaccharide Motif in N-Glycans through an Amide-Protection Strategy. 26929048 3 39 theme structure 545:553 arg1 synthesis 502:510 the synthesis 498:510 the synthesis of the as-described disialylated structure 498:553 This synthetic route enabled the synthesis of the as-described disialylated structure. 26929048 1 40 theme Neu5Ac 145:150 arg1 Gal 157:159 Neu5Ac(α2,3)Gal(β1,3) 145:165 Neu5Ac(α2,3)Gal(β1,3) 145:165 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 5 41 theme acid 918:921 arg1 residues 923:930 the sialic acid residues 907:930 the sialic acid residues 907:930 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 42 theme hydrogen 889:896 arg1 bonds 898:902 hydrogen bonds 889:902 hydrogen bonds 889:902 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 2 43 theme disaccharide 429:440 arg1 glycosylation 399:411 the glycosylation 395:411 the glycosylation of a sialylated disaccharide with a glucosaminyl donor 395:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 2 44 theme sialylated 418:427 arg1 disaccharide 429:440 a sialylated disaccharide 416:440 a sialylated disaccharide with a glucosaminyl donor 416:466 Compound 1 was obtained through an α-sialylation reaction between a sialic acid donor and a trisaccharide that was synthesized from the glycosylation of a sialylated disaccharide with a glucosaminyl donor. 26929048 5 45 attach presence 877:884 arg2 bonds 898:902 hydrogen bonds 889:902 hydrogen bonds 889:902 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 5 45 attach presence 877:884 arg1 residues 923:930 the sialic acid residues 907:930 the sialic acid residues 907:930 Protection of the amide groups in the sialic acid residues significantly increased the yield of the glycosylation reaction between the two sialylated disaccharides, thus suggesting that the presence of hydrogen bonds on the sialic acid residues diminished their reactivity. 26929048 1 46 located found 201:205 arg1 termini 214:220 the termini 210:220 the termini of some N-glycans 210:238 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 1 46 located found 201:205 arg2 which 192:196 which 192:196 which 192:196 A disialylated tetrasaccharide, Neu5Ac(α2,3)Gal(β1,3)[Neu5Ac(α2,6)]GlcNAc (1), which is found at the termini of some N-glycans, has been synthesized. 26929048 4 47 theme more-convergent 558:572 arg1 route 574:578 A more-convergent route 556:578 A more-convergent route based on the glycosylation of two sialylated disaccharides 556:637 A more-convergent route based on the glycosylation of two sialylated disaccharides was also established to scale up the synthesis. 24947385 5 0 theme proteins 806:813 arg1 endocytosis 783:793 endocytosis 783:793 endocytosis of various proteins 783:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 3 1 from role 445:448 arg1 internalization 477:491 the internalization 473:491 the internalization of insulin-responsive glucose transporter GLUT4 473:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 1 2 theme intracellular 152:164 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 2 theme intracellular 152:164 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 0 3 theme proteins 77:84 arg1 internalization 45:59 the internalization 41:59 the internalization of glycosylated proteins 41:84 Use of transmembrane FRET to investigate the internalization of glycosylated proteins. 24947385 3 4 theme transporter 523:533 arg1 GLUT4 535:539 insulin-responsive glucose transporter GLUT4 496:539 insulin-responsive glucose transporter GLUT4 496:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 5 5 theme glycan 759:764 arg1 modifications 766:778 specific glycan modifications 750:778 specific glycan modifications 750:778 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 5 6 theme glycan 695:700 arg1 tool 710:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 2 7 theme specific 337:344 arg1 modification 353:364 a specific glycan modification 335:364 a specific glycan modification 335:364 Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. 24947385 4 8 gly sialylated 556:565 arg1 glycoforms 567:576 sialylated glycoforms 556:576 sialylated glycoforms of GFP-tagged GLUT4 556:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 4 8 gly sialylated 556:565 arg1 GLUT4 592:596 GFP-tagged GLUT4 581:596 GFP-tagged GLUT4 581:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 5 9 theme modifications 766:778 arg1 roles 741:745 probing functional roles 722:745 probing functional roles of specific glycan modifications in endocytosis of various proteins 722:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 0 10 gly glycosylated 64:75 arg1 proteins 77:84 glycosylated proteins 64:84 glycosylated proteins 64:84 Use of transmembrane FRET to investigate the internalization of glycosylated proteins. 24947385 3 11 theme glucose 515:521 arg1 transporter 523:533 insulin-responsive glucose transporter 496:533 insulin-responsive glucose transporter GLUT4 496:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 5 12 from roles 741:745 arg1 endocytosis 783:793 endocytosis 783:793 endocytosis of various proteins 783:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 4 13 gly non-sialylated 641:654 arg1 GLUT4 656:660 non-sialylated GLUT4 641:660 non-sialylated GLUT4 641:660 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 2 14 theme novel 267:271 arg1 method 273:278 a novel method 265:278 a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification 265:364 Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. 24947385 1 15 theme intercellular 170:182 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 15 theme intercellular 170:182 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 3 16 dep transfer 405:412 arg1 technique 414:422 technique 414:422 a fluorescence resonance energy transfer technique 373:422 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 1 17 theme numerous 230:237 arg1 studies 239:245 numerous studies 230:245 numerous studies 230:245 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 3 18 theme fluorescence 375:386 arg1 transfer 405:412 a fluorescence resonance energy transfer 373:412 a fluorescence resonance energy transfer technique 373:422 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 4 19 theme GFP-tagged 581:590 arg1 GLUT4 592:596 GFP-tagged GLUT4 581:596 GFP-tagged GLUT4 581:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 2 20 theme proteins 306:313 arg1 endocytosis 291:301 endocytosis 291:301 endocytosis of proteins of interest bearing a specific glycan modification 291:364 Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. 24947385 3 21 theme resonance 388:396 arg1 transfer 405:412 a fluorescence resonance energy transfer 373:412 a fluorescence resonance energy transfer technique 373:422 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 1 22 theme proteins 199:206 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 22 theme proteins 199:206 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 0 23 theme FRET 21:24 arg1 Use 0:2 Use 0:2 Use of transmembrane FRET 0:24 Use of transmembrane FRET to investigate the internalization of glycosylated proteins. 24947385 5 24 theme functional 730:739 arg1 roles 741:745 probing functional roles 722:745 probing functional roles of specific glycan modifications in endocytosis of various proteins 722:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 3 25 theme structure 460:468 arg1 role 445:448 the role 441:448 the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4 441:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 0 26 theme transmembrane 7:19 arg1 FRET 21:24 transmembrane FRET 7:24 transmembrane FRET 7:24 Use of transmembrane FRET to investigate the internalization of glycosylated proteins. 24947385 4 27 theme GLUT4 592:596 arg1 glycoforms 567:576 sialylated glycoforms 556:576 sialylated glycoforms of GFP-tagged GLUT4 556:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 1 28 theme glycans 105:111 arg1 importance 91:100 The importance 87:100 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, 87:207 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 3 29 theme GLUT4 535:539 arg1 internalization 477:491 the internalization 473:491 the internalization of insulin-responsive glucose transporter GLUT4 473:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 3 30 theme insulin-responsive 496:513 arg1 transporter 523:533 insulin-responsive glucose transporter 496:533 insulin-responsive glucose transporter GLUT4 496:539 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 5 31 theme specific 750:757 arg1 modifications 766:778 specific glycan modifications 750:778 specific glycan modifications 750:778 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 4 32 gly glycoforms 567:576 arg1 GLUT4 592:596 GFP-tagged GLUT4 581:596 GFP-tagged GLUT4 581:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 3 33 theme glycan 453:458 arg1 structure 460:468 glycan structure 453:468 glycan structure 453:468 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 3 34 theme energy 398:403 arg1 transfer 405:412 a fluorescence resonance energy transfer 373:412 a fluorescence resonance energy transfer technique 373:422 Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. 24947385 5 35 theme imaging 702:708 arg1 tool 710:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 4 36 theme sialylated 556:565 arg1 glycoforms 567:576 sialylated glycoforms 556:576 sialylated glycoforms of GFP-tagged GLUT4 556:596 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 5 37 theme novel 689:693 arg1 tool 710:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool 684:713 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 1 38 theme various 116:122 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 38 theme various 116:122 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 39 theme cellular 124:131 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 39 theme cellular 124:131 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 5 40 theme probing 722:728 arg1 roles 741:745 probing functional roles 722:745 probing functional roles of specific glycan modifications in endocytosis of various proteins 722:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 24947385 2 41 theme interest 318:325 arg1 proteins 306:313 proteins 306:313 proteins of interest bearing a specific glycan modification 306:364 Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. 24947385 0 42 theme glycosylated 64:75 arg1 proteins 77:84 glycosylated proteins 64:84 glycosylated proteins 64:84 Use of transmembrane FRET to investigate the internalization of glycosylated proteins. 24947385 4 43 theme insulin 667:673 arg1 removal 675:681 insulin removal 667:681 insulin removal 667:681 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 2 44 theme glycan 346:351 arg1 modification 353:364 a specific glycan modification 335:364 a specific glycan modification 335:364 Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. 24947385 1 45 from importance 91:100 arg1 events 133:138 various cellular events 116:138 various cellular events 116:138 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 1 45 from importance 91:100 arg1 trafficking 184:194 intracellular and intercellular trafficking 152:194 trafficking 184:194 The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. 24947385 4 46 theme non-sialylated 641:654 arg1 GLUT4 656:660 non-sialylated GLUT4 641:660 non-sialylated GLUT4 641:660 We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. 24947385 5 47 theme various 798:804 arg1 proteins 806:813 various proteins 798:813 various proteins 798:813 This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins. 29136858 8 0 theme Hydrophilic 1241:1251 arg1 Chromatography 1265:1278 Hydrophilic Interaction Chromatography 1241:1278 Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans 1241:1315 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 1 1 theme major 187:191 arg1 challenge 193:201 a major challenge 185:201 a major challenge for analytical sciences 185:225 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 9 2 theme glycosylation 1558:1570 arg1 heterogeneity 1572:1584 the glycosylation heterogeneity 1554:1584 the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision 1554:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 7 3 theme reproducibility 1028:1042 arg1 terms 1004:1008 terms 1004:1008 terms of robustness and reproducibility 1004:1042 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 2 4 theme comprehensive 381:393 arg1 identification 395:408 comprehensive identification 381:408 comprehensive identification 381:408 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 4 5 theme analytical 622:631 arg1 methods 633:639 several analytical methods 614:639 several analytical methods 614:639 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 29136858 4 5 theme analytical 622:631 arg1 methods 678:684 separation-based or MS-based methods 649:684 separation-based or MS-based methods 649:684 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 29136858 0 6 theme method 114:119 arg1 validation 121:130 method validation 114:130 method validation 114:130 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 9 7 gly glycosylation 1558:1570 arg1 range 1596:1600 a wide range 1589:1600 a wide range of therapeutic mAbs with high accuracy and precision 1589:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 6 8 theme mAbs 865:868 arg1 characterization 845:860 the comprehensive characterization 827:860 the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species 827:963 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 7 9 from Validation 966:975 arg1 terms 1004:1008 terms 1004:1008 terms of robustness and reproducibility 1004:1042 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 5 10 theme CE-ESI-MS 715:723 arg1 approach 725:732 no CE-ESI-MS approach 712:732 no CE-ESI-MS approach 712:732 However, no CE-ESI-MS approach has been assessed and validated. 29136858 7 11 theme robustness 1013:1022 arg1 terms 1004:1008 terms 1004:1008 terms of robustness and reproducibility 1004:1042 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 3 12 theme glycosylation 538:550 arg1 consequence 525:535 a consequence 523:535 a consequence 523:535 As a consequence, glycosylation profiling must be deeply characterized. 29136858 3 12 theme glycosylation 538:550 arg1 profiling 552:560 glycosylation profiling 538:560 glycosylation profiling 538:560 As a consequence, glycosylation profiling must be deeply characterized. 29136858 6 13 gly glycopeptide 893:904 arg2 glycopeptide 893:904 the glycopeptide level 889:910 the glycopeptide level 889:910 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 8 14 theme glycans 1309:1315 arg1 Chromatography 1265:1278 Hydrophilic Interaction Chromatography 1241:1278 Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans 1241:1315 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 8 15 theme 2-aminobenzamide 1283:1298 arg1 glycans 1309:1315 2-aminobenzamide labelled glycans 1283:1315 2-aminobenzamide labelled glycans 1283:1315 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 6 16 theme species 957:963 arg1 quantitation 932:943 relative quantitation 923:943 relative quantitation of N-glycan species 923:963 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 8 17 theme Interaction 1253:1263 arg1 Chromatography 1265:1278 Hydrophilic Interaction Chromatography 1241:1278 Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans 1241:1315 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 9 18 theme wide 1591:1594 arg1 range 1596:1600 a wide range 1589:1600 a wide range of therapeutic mAbs with high accuracy and precision 1589:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 9 19 theme mAbs 1617:1620 arg1 range 1596:1600 a wide range 1589:1600 a wide range of therapeutic mAbs with high accuracy and precision 1589:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 6 20 theme N-glycan 948:955 arg1 species 957:963 N-glycan species 948:963 N-glycan species 948:963 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 6 21 theme comprehensive 831:843 arg1 characterization 845:860 the comprehensive characterization 827:860 the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species 827:963 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 0 22 theme Monoclonal 0:9 arg1 profiling 36:44 Monoclonal antibody N-glycosylation profiling 0:44 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry 0:96 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 1 23 theme analytical 207:216 arg1 sciences 218:225 analytical sciences 207:225 analytical sciences 207:225 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 0 24 theme antibody 11:18 arg1 profiling 36:44 Monoclonal antibody N-glycosylation profiling 0:44 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry 0:96 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 8 25 theme labelled 1300:1307 arg1 glycans 1309:1315 2-aminobenzamide labelled glycans 1283:1315 2-aminobenzamide labelled glycans 1283:1315 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 0 26 dep Assessment 99:108 arg1 profiling 36:44 Monoclonal antibody N-glycosylation profiling 0:44 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry 0:96 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 2 27 theme major 427:431 arg1 influence 433:441 their major influence 421:441 their major influence on protein structure and effector functions of monoclonal antibodies (mAbs) 421:517 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 2 28 theme monoclonal 490:499 arg1 mAbs 513:516 mAbs 513:516 mAbs 513:516 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 2 28 theme monoclonal 490:499 arg1 antibodies 501:510 monoclonal antibodies 490:510 monoclonal antibodies (mAbs) 490:517 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 7 29 theme method 994:999 arg1 Validation 966:975 Validation 966:975 Validation of the CE-ESI-MS method in terms of robustness and reproducibility 966:1042 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 9 30 theme therapeutic 1605:1615 arg1 mAbs 1617:1620 therapeutic mAbs 1605:1620 therapeutic mAbs with high accuracy and precision 1605:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 8 31 theme fluorescence 1322:1333 arg1 HILIC-FD 1345:1352 HILIC-FD 1345:1352 HILIC-FD 1345:1352 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 8 31 theme fluorescence 1322:1333 arg1 detector 1335:1342 fluorescence detector 1322:1342 fluorescence detector (HILIC-FD) analysis considered as a reference method 1322:1395 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 7 32 theme different 1156:1164 arg1 lines 1171:1175 different cell lines 1156:1175 different cell lines 1156:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 6 33 theme method 812:817 arg1 use 795:797 the use 791:797 the use of CE-ESI-MS method 791:817 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 2 34 theme effector 468:475 arg1 functions 477:485 effector functions 468:485 effector functions 468:485 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 0 35 theme N-glycosylation 20:34 arg1 profiling 36:44 Monoclonal antibody N-glycosylation profiling 0:44 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry 0:96 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 4 36 theme several 614:620 arg1 methods 633:639 several analytical methods 614:639 several analytical methods 614:639 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 29136858 4 36 theme several 614:620 arg1 methods 678:684 separation-based or MS-based methods 649:684 separation-based or MS-based methods 649:684 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 29136858 7 37 theme cell 1166:1169 arg1 lines 1171:1175 different cell lines 1156:1175 different cell lines 1156:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 8 38 theme reference 1380:1388 arg1 method 1390:1395 a reference method 1378:1395 a reference method 1378:1395 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 0 39 theme capillary 52:60 arg1 electrophoresis 62:76 capillary electrophoresis 52:76 capillary electrophoresis 52:76 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 9 40 with mAbs 1617:1620 arg1 precision 1645:1653 precision 1645:1653 precision 1645:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 9 40 with mAbs 1617:1620 arg1 accuracy 1632:1639 high accuracy 1627:1639 high accuracy 1627:1639 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 6 41 theme relative 923:930 arg1 quantitation 932:943 relative quantitation 923:943 relative quantitation of N-glycan species 923:963 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 6 42 dep mAbs 865:868 arg1 N-glycosylation 870:884 N-glycosylation 870:884 N-glycosylation 870:884 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 9 43 theme high 1627:1630 arg1 accuracy 1632:1639 high accuracy 1627:1639 high accuracy 1627:1639 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 8 44 theme detector 1335:1342 arg1 analysis 1355:1362 fluorescence detector (HILIC-FD) analysis 1322:1362 fluorescence detector (HILIC-FD) analysis considered as a reference method 1322:1395 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 9 45 dep CE-ESI-MS 1449:1457 arg1 the 1445:1447 the 1445:1447 the 1445:1447 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 8 46 theme Glycosylation 1178:1190 arg1 patterns 1192:1199 Glycosylation patterns 1178:1199 Glycosylation patterns obtained for each mAbs 1178:1222 Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. 29136858 6 47 theme CE-ESI-MS 802:810 arg1 method 812:817 CE-ESI-MS method 802:817 CE-ESI-MS method 802:817 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 2 48 theme protein 446:452 arg1 structure 454:462 protein structure 446:462 protein structure 446:462 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 6 49 from level 906:910 arg1 N-glycosylation 870:884 N-glycosylation 870:884 N-glycosylation 870:884 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 6 50 gly N-glycosylation 870:884 arg1 level 906:910 the glycopeptide level 889:910 the glycopeptide level 889:910 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 6 50 gly N-glycosylation 870:884 arg2 level 906:910 the glycopeptide level 889:910 the glycopeptide level 889:910 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 7 51 theme different 1129:1137 arg1 mAbs 1139:1142 ten different mAbs 1125:1142 ten different mAbs produced in different cell lines 1125:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 7 52 theme profiles 1112:1119 arg1 quantitation 1082:1093 the relative quantitation 1069:1093 the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines 1069:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 4 53 theme separation-based 649:664 arg1 methods 678:684 separation-based or MS-based methods 649:684 separation-based or MS-based methods 649:684 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 29136858 9 54 theme method 1518:1523 arg1 attractiveness 1490:1503 the attractiveness 1486:1503 the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision 1486:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 9 55 theme similar 1403:1409 arg1 glycoprofiling 1411:1424 Very similar glycoprofiling 1398:1424 Very similar glycoprofiling 1398:1424 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 6 56 theme glycopeptide 893:904 arg1 level 906:910 the glycopeptide level 889:910 the glycopeptide level 889:910 Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. 29136858 9 57 gly heterogeneity 1572:1584 arg1 range 1596:1600 a wide range 1589:1600 a wide range of therapeutic mAbs with high accuracy and precision 1589:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 2 58 from influence 433:441 arg1 structure 454:462 protein structure 446:462 protein structure 446:462 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 2 58 from influence 433:441 arg1 functions 477:485 effector functions 468:485 effector functions 468:485 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 1 59 theme therapeutic 153:163 arg1 proteins 165:172 therapeutic proteins 153:172 therapeutic proteins 153:172 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 7 60 theme CE-ESI-MS 984:992 arg1 method 994:999 the CE-ESI-MS method 980:999 the CE-ESI-MS method 980:999 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 7 61 theme glycosylation 1098:1110 arg1 profiles 1112:1119 glycosylation profiles 1098:1119 glycosylation profiles for ten different mAbs produced in different cell lines 1098:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 1 62 theme proteins 165:172 arg1 Characterization 133:148 Characterization 133:148 Characterization of therapeutic proteins 133:172 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 1 63 theme post-translational 264:281 arg1 modifications 283:295 post-translational modifications 264:295 post-translational modifications (PTM) 264:301 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 1 63 theme post-translational 264:281 arg1 PTM 298:300 PTM 298:300 PTM 298:300 Characterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). 29136858 7 64 theme relative 1073:1080 arg1 quantitation 1082:1093 the relative quantitation 1069:1093 the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines 1069:1175 Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. 29136858 9 65 theme range 1596:1600 arg1 heterogeneity 1572:1584 the glycosylation heterogeneity 1554:1584 the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision 1554:1653 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 0 66 theme Mass 80:83 arg1 spectrometry 85:96 Mass spectrometry 80:96 Mass spectrometry 80:96 Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. 29136858 9 67 theme CE-ESI-MS 1508:1516 arg1 method 1518:1523 CE-ESI-MS method 1508:1523 CE-ESI-MS method 1508:1523 Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision. 29136858 2 68 theme antibodies 501:510 arg1 structure 454:462 protein structure 446:462 protein structure 446:462 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 2 68 theme antibodies 501:510 arg1 functions 477:485 effector functions 468:485 effector functions 468:485 Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). 29136858 4 69 theme MS-based 669:676 arg1 methods 678:684 separation-based or MS-based methods 649:684 separation-based or MS-based methods 649:684 For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. 28534482 6 0 theme deeper 1125:1130 arg1 coverage 1132:1139 a deeper coverage 1123:1139 a deeper coverage (compared to lectin enrichment) 1123:1171 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 1 1 gly glycopeptides 182:194 arg2 glycopeptides 182:194 N-linked glycopeptides 173:194 N-linked glycopeptides 173:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 6 2 dep coverage 1132:1139 arg1 compared 1142:1149 compared 1142:1149 compared to lectin enrichment 1142:1170 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 3 3 theme diverse 531:537 arg1 types 539:543 diverse types 531:543 diverse types of N-linked glycomolecules 531:570 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 6 4 gly N-glycopeptide 1005:1018 arg2 N-glycopeptide 1005:1018 this highly efficient N-glycopeptide enrichment method 983:1036 this highly efficient N-glycopeptide enrichment method 983:1036 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 2 5 theme pentasaccharide 434:448 arg1 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 5 theme pentasaccharide 434:448 arg1 motif 450:454 the common core pentasaccharide motif 418:454 the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins 418:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 5 6 link N-linked 924:931 arg1 glycopeptides 933:945 N-linked glycopeptides 924:945 N-linked glycopeptides from human serum 924:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 2 7 link N-linked 473:480 arg1 glycoproteins 482:494 N-linked glycoproteins 473:494 N-linked glycoproteins 473:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 8 theme degradation 386:396 arg1 system 398:403 the ubiquitin-mediated degradation system 363:403 the ubiquitin-mediated degradation system 363:403 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 6 9 theme composition 1089:1099 arg1 determination 1063:1075 the simultaneous determination 1046:1075 the simultaneous determination of N-glycan composition and N-glycosites 1046:1116 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 6 10 theme N-glycosites 1105:1116 arg1 determination 1063:1075 the simultaneous determination 1046:1075 the simultaneous determination of N-glycan composition and N-glycosites 1046:1116 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 1 11 gly micro-heterogeneity 237:255 arg1 N-glycosylation 260:274 N-glycosylation 260:274 N-glycosylation 260:274 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 2 12 theme properties 320:329 arg1 advantage 294:302 advantage 294:302 advantage 294:302 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 0 13 theme N-glycans 87:95 arg1 capture 76:82 selective capture 66:82 selective capture of N-glycans and N-glycopeptides 66:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 3 14 theme wild-type 582:590 arg1 Fbs1 592:595 wild-type Fbs1 582:595 wild-type Fbs1 582:595 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 2 15 theme inherent 311:318 arg1 properties 320:329 the inherent properties 307:329 the inherent properties of Fbs1 307:337 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 4 16 theme complex 785:791 arg1 N-glycomolecules 793:808 complex N-glycomolecules 785:808 complex N-glycomolecules 785:808 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 6 17 theme large-scale 1186:1196 arg1 studies 1216:1222 large-scale N-glycoproteomics studies 1186:1222 large-scale N-glycoproteomics studies 1186:1222 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 3 18 theme glycomolecules 557:570 arg1 types 539:543 diverse types 531:543 diverse types of N-linked glycomolecules 531:570 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 5 19 theme human 952:956 arg1 serum 958:962 human serum 952:962 human serum 952:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 1 20 theme N-linked 173:180 arg1 glycopeptides 182:194 N-linked glycopeptides 173:194 N-linked glycopeptides 173:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 0 21 theme N-glycopeptides 101:115 arg1 capture 76:82 selective capture 66:82 selective capture of N-glycans and N-glycopeptides 66:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 5 22 theme Fbs1 850:853 arg1 variant 859:865 the Fbs1 GYR variant 846:865 the Fbs1 GYR variant 846:865 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 2 23 theme core 429:432 arg1 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 23 theme core 429:432 arg1 motif 450:454 the common core pentasaccharide motif 418:454 the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins 418:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 3 24 theme N-linked 548:555 arg1 glycomolecules 557:570 N-linked glycomolecules 548:570 N-linked glycomolecules 548:570 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 1 25 link N-linked 173:180 arg1 glycopeptides 182:194 N-linked glycopeptides 173:194 N-linked glycopeptides 173:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 2 26 theme common 422:427 arg1 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 26 theme common 422:427 arg1 motif 450:454 the common core pentasaccharide motif 418:454 the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins 418:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 27 theme glycoproteins 482:494 arg1 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 Man3GlcNAc2 457:467 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 2 27 theme glycoproteins 482:494 arg1 motif 450:454 the common core pentasaccharide motif 418:454 the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins 418:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 6 28 theme N-glycan 1080:1087 arg1 composition 1089:1099 N-glycan composition 1080:1099 N-glycan composition 1080:1099 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 5 29 theme glycopeptides 933:945 arg1 enrichment 910:919 substantially unbiased enrichment 887:919 substantially unbiased enrichment of N-linked glycopeptides from human serum 887:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 5 30 gly glycopeptides 933:945 arg2 glycopeptides 933:945 N-linked glycopeptides 924:945 N-linked glycopeptides from human serum 924:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 5 30 gly glycopeptides 933:945 arg1 serum 958:962 human serum 952:962 human serum 952:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 0 31 theme affinity 19:26 arg1 protein 54:60 high affinity Fbs1 carbohydrate binding protein 14:60 high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides 14:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 0 32 gly N-glycopeptides 101:115 arg2 N-glycopeptides 101:115 N-glycopeptides 101:115 N-glycopeptides 101:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 5 33 theme unbiased 901:908 arg1 enrichment 910:919 substantially unbiased enrichment 887:919 substantially unbiased enrichment of N-linked glycopeptides from human serum 887:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 0 34 theme high 14:17 arg1 protein 54:60 high affinity Fbs1 carbohydrate binding protein 14:60 high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides 14:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 4 35 theme Fbs1 665:668 arg1 variants 670:677 Fbs1 variants 665:677 Fbs1 variants 665:677 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 6 36 theme N-glycoproteomics 1198:1214 arg1 studies 1216:1222 large-scale N-glycoproteomics studies 1186:1222 large-scale N-glycoproteomics studies 1186:1222 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 6 37 theme efficient 995:1003 arg1 method 1031:1036 this highly efficient N-glycopeptide enrichment method 983:1036 this highly efficient N-glycopeptide enrichment method 983:1036 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 6 38 theme lectin 1154:1159 arg1 enrichment 1161:1170 lectin enrichment 1154:1170 lectin enrichment 1154:1170 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 0 39 theme carbohydrate 33:44 arg1 protein 54:60 high affinity Fbs1 carbohydrate binding protein 14:60 high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides 14:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 6 40 theme simultaneous 1050:1061 arg1 determination 1063:1075 the simultaneous determination 1046:1075 the simultaneous determination of N-glycan composition and N-glycosites 1046:1116 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 4 41 theme display 711:717 arg1 selection 719:727 plasmid display selection 703:727 plasmid display selection 703:727 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 0 42 theme Fbs1 28:31 arg1 protein 54:60 high affinity Fbs1 carbohydrate binding protein 14:60 high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides 14:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 3 43 link N-linked 548:555 arg1 glycomolecules 557:570 N-linked glycomolecules 548:570 N-linked glycomolecules 548:570 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 2 44 theme N-linked 473:480 arg1 glycoproteins 482:494 N-linked glycoproteins 473:494 N-linked glycoproteins 473:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 6 45 gly N-glycosites 1105:1116 arg2 N-glycosites 1105:1116 N-glycosites 1105:1116 N-glycosites 1105:1116 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 4 46 theme plasmid 703:709 arg1 selection 719:727 plasmid display selection 703:727 plasmid display selection 703:727 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 3 47 theme high-mannose-containing 618:640 arg1 glycans 642:648 high-mannose-containing glycans 618:648 high-mannose-containing glycans 618:648 We show that Fbs1 is able to bind diverse types of N-linked glycomolecules; however, wild-type Fbs1 preferentially binds high-mannose-containing glycans. 28534482 2 48 theme ubiquitin-mediated 367:384 arg1 system 398:403 the ubiquitin-mediated degradation system 363:403 the ubiquitin-mediated degradation system 363:403 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 0 49 theme binding 46:52 arg1 protein 54:60 high affinity Fbs1 carbohydrate binding protein 14:60 high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides 14:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 6 50 theme sample 1247:1252 arg1 complexity 1254:1263 greatly reduced sample complexity 1231:1263 greatly reduced sample complexity 1231:1263 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 6 51 theme enrichment 1020:1029 arg1 method 1031:1036 this highly efficient N-glycopeptide enrichment method 983:1036 this highly efficient N-glycopeptide enrichment method 983:1036 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 4 52 contain possess 736:742 arg1 mutagenesis 687:697 mutagenesis 687:697 mutagenesis 687:697 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 4 52 contain possess 736:742 arg1 selection 719:727 plasmid display selection 703:727 plasmid display selection 703:727 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 4 52 contain possess 736:742 arg2 affinity 751:758 higher affinity 744:758 higher affinity 744:758 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 2 53 gly glycoproteins 482:494 arg1 glycoproteins 482:494 N-linked glycoproteins 473:494 N-linked glycoproteins 473:494 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 4 54 theme higher 744:749 arg1 affinity 751:758 higher affinity 744:758 higher affinity 744:758 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 4 55 theme N-glycomolecules 793:808 arg1 recovery 773:780 recovery 773:780 recovery of complex N-glycomolecules 773:808 We identified Fbs1 variants through mutagenesis and plasmid display selection, which possess higher affinity and improved recovery of complex N-glycomolecules. 28534482 2 56 theme Fbs1 334:337 arg1 properties 320:329 the inherent properties 307:329 the inherent properties of Fbs1 307:337 The method takes advantage of the inherent properties of Fbs1, which functions within the ubiquitin-mediated degradation system to recognize the common core pentasaccharide motif (Man3GlcNAc2) of N-linked glycoproteins. 28534482 1 57 theme micro-heterogeneity 237:255 arg1 detection 224:232 detection 224:232 detection of micro-heterogeneity of N-glycosylation 224:274 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 5 58 from enrichment 910:919 arg1 serum 958:962 human serum 952:962 human serum 952:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 1 59 theme selective 131:139 arg1 enrichment 159:168 selective and comprehensive enrichment 131:168 selective and comprehensive enrichment of N-linked glycopeptides 131:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 6 60 theme reduced 1239:1245 arg1 complexity 1254:1263 greatly reduced sample complexity 1231:1263 greatly reduced sample complexity 1231:1263 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 0 61 theme selective 66:74 arg1 capture 76:82 selective capture 66:82 selective capture of N-glycans and N-glycopeptides 66:115 An engineered high affinity Fbs1 carbohydrate binding protein for selective capture of N-glycans and N-glycopeptides. 28534482 5 62 theme N-linked 924:931 arg1 glycopeptides 933:945 N-linked glycopeptides 924:945 N-linked glycopeptides from human serum 924:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 5 63 from serum 958:962 arg1 enrichment 910:919 substantially unbiased enrichment 887:919 substantially unbiased enrichment of N-linked glycopeptides from human serum 887:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 5 63 from serum 958:962 arg1 glycopeptides 933:945 N-linked glycopeptides 924:945 N-linked glycopeptides from human serum 924:962 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 1 64 theme glycopeptides 182:194 arg1 enrichment 159:168 selective and comprehensive enrichment 131:168 selective and comprehensive enrichment of N-linked glycopeptides 131:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 1 65 theme N-glycosylation 260:274 arg1 micro-heterogeneity 237:255 micro-heterogeneity 237:255 micro-heterogeneity of N-glycosylation 237:274 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 28534482 6 66 theme N-glycopeptide 1005:1018 arg1 method 1031:1036 this highly efficient N-glycopeptide enrichment method 983:1036 this highly efficient N-glycopeptide enrichment method 983:1036 Most importantly, this highly efficient N-glycopeptide enrichment method enables the simultaneous determination of N-glycan composition and N-glycosites with a deeper coverage (compared to lectin enrichment) and improves large-scale N-glycoproteomics studies due to greatly reduced sample complexity. 28534482 5 67 theme GYR 855:857 arg1 variant 859:865 the Fbs1 GYR variant 846:865 the Fbs1 GYR variant 846:865 In particular, we demonstrate that the Fbs1 GYR variant may be employed for substantially unbiased enrichment of N-linked glycopeptides from human serum. 28534482 1 68 theme comprehensive 145:157 arg1 enrichment 159:168 selective and comprehensive enrichment 131:168 selective and comprehensive enrichment of N-linked glycopeptides 131:194 A method for selective and comprehensive enrichment of N-linked glycopeptides was developed to facilitate detection of micro-heterogeneity of N-glycosylation. 27423877 4 0 theme trisaccharide 621:633 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 0 theme trisaccharide 621:633 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 6 1 theme good 966:969 arg1 yields 971:976 good yields 966:976 good yields 966:976 The heavily branched hexasaccharide was obtained in good yields and excellent α-selectivity. 27423877 7 2 theme octasaccharide 1025:1038 arg1 motif 1057:1061 The frame shifted octasaccharide structural triad motif 1007:1061 The frame shifted octasaccharide structural triad motif for serotype C 1007:1076 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 2 3 theme polysaccharide 362:375 arg1 heterogeneity 332:344 the heterogeneity 328:344 the heterogeneity of the capsular polysaccharide 328:375 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 2 3 theme polysaccharide 362:375 arg1 difficulty 385:394 the difficulty 381:394 the difficulty of identifying protective epitopes 381:429 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 4 4 theme protected 611:619 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 4 theme protected 611:619 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 7 5 theme structural 1040:1049 arg1 motif 1057:1061 The frame shifted octasaccharide structural triad motif 1007:1061 The frame shifted octasaccharide structural triad motif for serotype C 1007:1076 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 2 6 theme open 306:309 arg1 development 215:225 The development 211:225 The development of a vaccine based on the capsular polysaccharide of C. neoformans 211:292 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 2 6 theme open 306:309 arg1 challenge 311:319 an open challenge 303:319 an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes 303:429 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 4 7 theme building 649:656 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 7 theme building 649:656 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 8 from polysaccharide 709:722 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 8 from polysaccharide 709:722 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 9 theme thioglycoside 635:647 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 9 theme thioglycoside 635:647 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 5 10 theme temporary 886:894 arg1 group 907:911 the temporary protecting group 882:911 the temporary protecting group 882:911 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 1 11 theme fungal 121:126 arg1 neoformans 105:114 Cryptococcus neoformans 92:114 Cryptococcus neoformans 92:114 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27423877 1 11 theme fungal 121:126 arg1 pathogen 128:135 a fungal pathogen 119:135 a fungal pathogen that can cause life-threatening infections in immunocompromised patients 119:208 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27423877 8 12 theme C. 1229:1230 arg1 serotypes 1243:1251 the four C. neoformans serotypes 1220:1251 the four C. neoformans serotypes 1220:1251 For the first time this structural motif, which is the most substituted amongst the four C. neoformans serotypes, was prepared. 27423877 5 13 with reaction 768:775 arg1 acceptor 790:797 a model acceptor 782:797 a model acceptor 782:797 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 1 14 theme immunocompromised 183:199 arg1 patients 201:208 immunocompromised patients 183:208 immunocompromised patients 183:208 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27423877 9 15 theme synthetic 1483:1491 arg1 vaccine 1493:1499 a synthetic vaccine 1481:1499 a synthetic vaccine against this pathogen 1481:1521 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 9 16 from penta-up 1343:1350 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 5 17 from donor 743:747 arg1 reaction 768:775 a glycosylation reaction 752:775 a glycosylation reaction with a model acceptor 752:797 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 3 18 theme capsule 521:527 arg1 structures 485:494 structurally defined part structures 459:494 structurally defined part structures of the C. neoformans GXM capsule 459:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 2 19 theme protective 411:420 arg1 epitopes 422:429 protective epitopes 411:429 protective epitopes 411:429 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 6 20 theme branched 926:933 arg1 hexasaccharide 935:948 The heavily branched hexasaccharide 914:948 The heavily branched hexasaccharide 914:948 The heavily branched hexasaccharide was obtained in good yields and excellent α-selectivity. 27423877 4 21 theme C 707:707 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 1 22 theme Cryptococcus 92:103 arg1 neoformans 105:114 Cryptococcus neoformans 92:114 Cryptococcus neoformans 92:114 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27423877 1 22 theme Cryptococcus 92:103 arg1 pathogen 128:135 a fungal pathogen 119:135 a fungal pathogen that can cause life-threatening infections in immunocompromised patients 119:208 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27423877 8 23 theme structural 1164:1173 arg1 most 1195:1198 most 1195:1198 most 1195:1198 For the first time this structural motif, which is the most substituted amongst the four C. neoformans serotypes, was prepared. 27423877 8 23 theme structural 1164:1173 arg1 motif 1175:1179 this structural motif 1159:1179 this structural motif 1159:1179 For the first time this structural motif, which is the most substituted amongst the four C. neoformans serotypes, was prepared. 27423877 7 24 theme block 1124:1128 arg1 strategy 1130:1137 the same building block strategy 1106:1137 the same building block strategy 1106:1137 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 5 25 theme group 907:911 arg1 removal 871:877 removal 871:877 removal of the temporary protecting group 871:911 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 0 26 dep polysaccharide 76:89 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of part structures of Cryptococcus neoformans 0:54 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 0 27 theme structures 18:27 arg1 Synthesis 0:8 Synthesis 0:8 Synthesis of part structures of Cryptococcus neoformans 0:54 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 8 28 theme first 1148:1152 arg1 time 1154:1157 the first time 1144:1157 the first time 1144:1157 For the first time this structural motif, which is the most substituted amongst the four C. neoformans serotypes, was prepared. 27423877 4 29 located present 673:679 arg2 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 29 located present 673:679 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 29 located present 673:679 arg2 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 2 30 theme due 321:323 arg1 development 215:225 The development 211:225 The development of a vaccine based on the capsular polysaccharide of C. neoformans 211:292 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 2 30 theme due 321:323 arg1 challenge 311:319 an open challenge 303:319 an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes 303:429 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 4 31 theme C. 684:685 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 0 32 theme part 13:16 arg1 structures 18:27 part structures 13:27 part structures of Cryptococcus neoformans 13:54 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 4 33 theme 3-O-naphthalenylmethyl 588:609 arg1 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 33 theme 3-O-naphthalenylmethyl 588:609 arg1 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 9 34 theme C. 1286:1287 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 3 35 theme great 535:539 arg1 demand 541:546 great demand 535:546 great demand 535:546 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 7 36 theme shifted 1017:1023 arg1 motif 1057:1061 The frame shifted octasaccharide structural triad motif 1007:1061 The frame shifted octasaccharide structural triad motif for serotype C 1007:1076 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 3 37 theme C. 503:504 arg1 capsule 521:527 the C. neoformans GXM capsule 499:527 the C. neoformans GXM capsule 499:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 0 38 theme Cryptococcus 32:43 arg1 neoformans 45:54 Cryptococcus neoformans 32:54 Cryptococcus neoformans 32:54 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 5 39 theme glycosylation 754:766 arg1 reaction 768:775 a glycosylation reaction 752:775 a glycosylation reaction with a model acceptor 752:797 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 4 40 theme serotype 698:705 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 2 41 theme capsular 253:260 arg1 polysaccharide 262:275 the capsular polysaccharide 249:275 the capsular polysaccharide of C. neoformans 249:292 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 3 42 theme structures 485:494 arg1 construction 443:454 construction 443:454 construction of structurally defined part structures of the C. neoformans GXM capsule 443:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 3 43 theme defined 472:478 arg1 structures 485:494 structurally defined part structures 459:494 structurally defined part structures of the C. neoformans GXM capsule 459:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 9 44 theme serotype 1300:1307 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 4 45 from present 673:679 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 5 46 theme protecting 896:905 arg1 group 907:911 the temporary protecting group 882:911 the temporary protecting group 882:911 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 0 47 theme serotype 56:63 arg1 polysaccharide 76:89 serotype C capsular polysaccharide 56:89 serotype C capsular polysaccharide 56:89 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 9 48 theme unique 1412:1417 arg1 glycoarrays 1419:1429 unique glycoarrays 1412:1429 unique glycoarrays 1412:1429 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 7 49 theme same 1110:1113 arg1 block 1124:1128 the same building block 1106:1128 the same building block strategy 1106:1137 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 9 50 theme C 1309:1309 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 4 51 dep C. 684:685 arg1 neoformans 687:696 neoformans 687:696 neoformans 687:696 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 9 52 theme synthesized 1274:1284 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 2 53 theme capsular 353:360 arg1 polysaccharide 362:375 the capsular polysaccharide 349:375 the capsular polysaccharide 349:375 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 0 54 theme neoformans 45:54 arg1 structures 18:27 part structures 13:27 part structures of Cryptococcus neoformans 13:54 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 7 55 theme building 1115:1122 arg1 block 1124:1128 the same building block 1106:1128 the same building block strategy 1106:1137 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 2 56 theme vaccine 232:238 arg1 challenge 311:319 an open challenge 303:319 an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes 303:429 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 2 56 theme vaccine 232:238 arg1 development 215:225 The development 211:225 The development of a vaccine based on the capsular polysaccharide of C. neoformans 211:292 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 4 57 attach present 673:679 arg2 present 673:679 present 673:679 present 673:679 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 57 attach present 673:679 arg1 polysaccharide 709:722 C. neoformans serotype C polysaccharide 684:722 C. neoformans serotype C polysaccharide 684:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 4 57 attach present 673:679 arg2 block 658:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block 586:662 a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 586:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 6 58 theme excellent 982:990 arg1 α-selectivity 992:1004 excellent α-selectivity 982:1004 excellent α-selectivity 982:1004 The heavily branched hexasaccharide was obtained in good yields and excellent α-selectivity. 27423877 2 59 gly heterogeneity 332:344 arg1 polysaccharide 362:375 the capsular polysaccharide 349:375 the capsular polysaccharide 349:375 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 2 60 theme neoformans 283:292 arg1 polysaccharide 262:275 the capsular polysaccharide 249:275 the capsular polysaccharide of C. neoformans 249:292 The development of a vaccine based on the capsular polysaccharide of C. neoformans is still an open challenge due to the heterogeneity of the capsular polysaccharide and the difficulty of identifying protective epitopes. 27423877 7 61 dep shifted 1017:1023 arg1 frame 1011:1015 frame 1011:1015 frame 1011:1015 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 5 62 theme model 784:788 arg1 acceptor 790:797 a model acceptor 782:797 a model acceptor 782:797 Its property as a donor in a glycosylation reaction with a model acceptor has been evaluated together with its behaviour as an acceptor following removal of the temporary protecting group. 27423877 9 63 theme varying 1324:1330 arg1 size 1332:1335 varying size 1324:1335 varying size 1324:1335 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 8 64 theme neoformans 1232:1241 arg1 serotypes 1243:1251 the four C. neoformans serotypes 1220:1251 the four C. neoformans serotypes 1220:1251 For the first time this structural motif, which is the most substituted amongst the four C. neoformans serotypes, was prepared. 27423877 3 65 theme GXM 517:519 arg1 capsule 521:527 the C. neoformans GXM capsule 499:527 the C. neoformans GXM capsule 499:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 4 66 theme block 658:662 arg1 synthesis 573:581 the synthesis 569:581 the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide 569:722 Herein is presented the synthesis of a 3-O-naphthalenylmethyl protected trisaccharide thioglycoside building block which is present in C. neoformans serotype C polysaccharide. 27423877 0 67 theme capsular 67:74 arg1 polysaccharide 76:89 serotype C capsular polysaccharide 56:89 serotype C capsular polysaccharide 56:89 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 9 68 theme size 1332:1335 arg1 fragments 1311:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments 1268:1319 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 7 69 theme serotype 1067:1074 arg1 C 1076:1076 serotype C 1067:1076 serotype C 1067:1076 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 3 70 theme part 480:483 arg1 structures 485:494 structurally defined part structures 459:494 structurally defined part structures of the C. neoformans GXM capsule 459:527 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 9 71 dep C. 1286:1287 arg1 neoformans 1289:1298 neoformans 1289:1298 neoformans 1289:1298 Three synthesized C. neoformans serotype C fragments of varying size, from penta-up to octasaccharide, were deprotected and will be included in unique glycoarrays to further investigate the possibility to develop a synthetic vaccine against this pathogen. 27423877 0 72 theme C 65:65 arg1 polysaccharide 76:89 serotype C capsular polysaccharide 56:89 serotype C capsular polysaccharide 56:89 Synthesis of part structures of Cryptococcus neoformans serotype C capsular polysaccharide. 27423877 3 73 dep C. 503:504 arg1 neoformans 506:515 neoformans 506:515 neoformans 506:515 Therefore, construction of structurally defined part structures of the C. neoformans GXM capsule is in great demand. 27423877 7 74 theme triad 1051:1055 arg1 motif 1057:1061 The frame shifted octasaccharide structural triad motif 1007:1061 The frame shifted octasaccharide structural triad motif for serotype C 1007:1076 The frame shifted octasaccharide structural triad motif for serotype C was also prepared following the same building block strategy. 27423877 1 75 theme life-threatening 152:167 arg1 infections 169:178 life-threatening infections 152:178 life-threatening infections 152:178 Cryptococcus neoformans is a fungal pathogen that can cause life-threatening infections in immunocompromised patients. 27705835 2 0 theme type 419:422 arg1 N-glycans 430:438 Lewis a type plant N-glycans 411:438 Lewis a type plant N-glycans 411:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 5 1 theme Lewis 1062:1066 arg1 type 1070:1073 the Lewis a type 1058:1073 the Lewis a type 1058:1073 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 2 theme a 172:172 arg1 epitope 174:180 Lewis a epitope 166:180 Lewis a epitope 166:180 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 2 3 from significance 395:406 arg1 system 478:483 the human immune system 461:483 the human immune system 461:483 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 2 4 theme a 417:417 arg1 N-glycans 430:438 Lewis a type plant N-glycans 411:438 Lewis a type plant N-glycans 411:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 1 5 theme plant 297:301 arg1 N-glycans 316:324 plant complex type N-glycans 297:324 plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-) 297:377 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 0 6 theme Lewis 166:170 arg1 epitope 174:180 Lewis a epitope 166:180 Lewis a epitope 166:180 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 3 7 theme N-glycans 560:568 arg1 N-glycans 560:568 such plant specific N-glycans 540:568 such plant specific N-glycans 540:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 3 7 theme N-glycans 560:568 arg1 amount 530:535 a substantial amount 516:535 a substantial amount of such plant specific N-glycans 516:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 5 8 theme substantial 1179:1189 arg1 amounts 1191:1197 substantial amounts 1179:1197 substantial amounts 1179:1197 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 9 theme plants 91:96 arg1 kinds 76:80 three kinds 70:80 three kinds of water plants 70:96 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 1 10 theme complex 303:309 arg1 N-glycans 316:324 plant complex type N-glycans 297:324 plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-) 297:377 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 5 11 theme N-glycans 912:920 arg1 analysis 896:903 Structural analysis 885:903 Structural analysis of the N-glycans 885:920 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 12 theme water 85:89 arg1 plants 91:96 water plants 85:96 water plants 85:96 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 1 13 theme type 311:314 arg1 N-glycans 316:324 plant complex type N-glycans 297:324 plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-) 297:377 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 5 14 theme plant 1160:1164 arg1 N-glycans 1166:1174 Lewis a type plant N-glycans 1147:1174 Lewis a type plant N-glycans 1147:1174 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 15 theme Structural 885:894 arg1 analysis 896:903 Structural analysis 885:903 Structural analysis of the N-glycans 885:920 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 1 16 theme mountain 235:242 arg1 allergen 257:264 the mountain cedar pollen allergen 231:264 the mountain cedar pollen allergen (Jun a1) 231:273 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 16 theme mountain 235:242 arg1 a1 271:272 Jun a1 267:272 Jun a1 267:272 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 0 17 theme Predominant 99:109 arg1 occurrence 111:120 Predominant occurrence 99:120 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 0:181 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 5 18 theme type 1033:1036 arg1 N-glycans 1038:1046 plant complex type N-glycans 1019:1046 plant complex type N-glycans 1019:1046 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 4 19 theme water 766:770 arg1 plants 772:777 three water plants 760:777 three water plants 760:777 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 4 19 theme water 766:770 arg1 nuttallii 787:795 Elodea nuttallii 780:795 Elodea nuttallii 780:795 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 4 19 theme water 766:770 arg1 densa 805:809 Egeria densa 798:809 Egeria densa 798:809 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 4 19 theme water 766:770 arg1 demersum 830:837 Ceratophyllum demersum 816:837 Ceratophyllum demersum 816:837 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 1 20 gly glycosylated 279:290 arg1 allergen 257:264 the mountain cedar pollen allergen 231:264 the mountain cedar pollen allergen (Jun a1) 231:273 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 20 gly glycosylated 279:290 arg1 Cry 219:221 Cry 219:221 Cry 219:221 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 20 gly glycosylated 279:290 arg1 allergen 209:216 The Japanese cedar pollen allergen 183:216 The Japanese cedar pollen allergen (Cry j1) 183:225 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 20 gly glycosylated 279:290 arg1 a1 271:272 Jun a1 267:272 Jun a1 267:272 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 5 21 theme plant 1019:1023 arg1 N-glycans 1038:1046 plant complex type N-glycans 1019:1046 plant complex type N-glycans 1019:1046 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 4 22 theme a 876:876 arg1 units 878:882 Lewis a units 870:882 Lewis a units 870:882 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 0 23 attach linked 33:38 arg1 glycoproteins 43:55 glycoproteins 43:55 glycoproteins expressed in three kinds of water plants 43:96 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 0 23 attach linked 33:38 arg2 N-glycans 23:31 N-glycans 23:31 N-glycans linked to glycoproteins expressed in three kinds of water plants 23:96 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 5 24 contain carry 1013:1017 arg1 glycoproteins 947:959 almost all glycoproteins 936:959 almost all glycoproteins expressed in these three water plants 936:997 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 24 contain carry 1013:1017 arg2 N-glycans 1038:1046 plant complex type N-glycans 1019:1046 plant complex type N-glycans 1019:1046 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 3 25 theme good 640:643 arg1 sources 658:664 good plant-glycan sources 640:664 good plant-glycan sources 640:664 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 3 26 theme Lewis 682:686 arg1 N-glycans 715:723 Lewis a epitope-containing plant N-glycans 682:723 Lewis a epitope-containing plant N-glycans 682:723 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 4 27 theme Lewis 870:874 arg1 units 878:882 Lewis a units 870:882 Lewis a units 870:882 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 0 28 theme Structural 0:9 arg1 features 11:18 Structural features 0:18 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 0:181 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 1 29 theme cedar 244:248 arg1 allergen 257:264 the mountain cedar pollen allergen 231:264 the mountain cedar pollen allergen (Jun a1) 231:273 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 29 theme cedar 244:248 arg1 a1 271:272 Jun a1 267:272 Jun a1 267:272 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 5 30 theme water 986:990 arg1 plants 992:997 these three water plants 974:997 these three water plants 974:997 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 31 theme N-glycans 23:31 arg1 features 11:18 Structural features 0:18 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 0:181 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 0 32 dep features 11:18 arg1 occurrence 111:120 Predominant occurrence 99:120 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 0:181 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 5 33 from preparation 1132:1142 arg1 amounts 1191:1197 substantial amounts 1179:1197 substantial amounts 1179:1197 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 34 gly glycoproteins 947:959 arg1 glycoproteins 947:959 almost all glycoproteins 936:959 almost all glycoproteins expressed in these three water plants 936:997 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 2 35 theme immune 471:476 arg1 system 478:483 the human immune system 461:483 the human immune system 461:483 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 1 36 theme Japanese 187:194 arg1 Cry 219:221 Cry 219:221 Cry 219:221 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 36 theme Japanese 187:194 arg1 allergen 209:216 The Japanese cedar pollen allergen 183:216 The Japanese cedar pollen allergen (Cry j1) 183:225 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 2 37 theme Lewis 411:415 arg1 N-glycans 430:438 Lewis a type plant N-glycans 411:438 Lewis a type plant N-glycans 411:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 5 38 theme good 1115:1118 arg1 plants 1104:1109 these water plants 1092:1109 these water plants 1092:1109 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 38 theme good 1115:1118 arg1 sources 1120:1126 good sources 1115:1126 good sources for preparation of Lewis a type plant N-glycans in substantial amounts 1115:1197 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 39 theme plant 129:133 arg1 N-glycans 148:156 the plant complex type N-glycans 125:156 the plant complex type N-glycans bearing Lewis a epitope 125:180 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 1 40 theme epitope 342:348 arg1 s 350:350 a epitope(s) 340:351 Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-) 334:377 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 3 41 theme plant-glycan 645:656 arg1 sources 658:664 good plant-glycan sources 640:664 good plant-glycan sources 640:664 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 1 42 theme cedar 196:200 arg1 Cry 219:221 Cry 219:221 Cry 219:221 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 42 theme cedar 196:200 arg1 allergen 209:216 The Japanese cedar pollen allergen 183:216 The Japanese cedar pollen allergen (Cry j1) 183:225 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 3 43 theme a 688:688 arg1 N-glycans 715:723 Lewis a epitope-containing plant N-glycans 682:723 Lewis a epitope-containing plant N-glycans 682:723 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 1 44 theme Jun 267:269 arg1 allergen 257:264 the mountain cedar pollen allergen 231:264 the mountain cedar pollen allergen (Jun a1) 231:273 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 44 theme Jun 267:269 arg1 a1 271:272 Jun a1 267:272 Jun a1 267:272 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 3 45 theme specific 551:558 arg1 N-glycans 560:568 such plant specific N-glycans 540:568 such plant specific N-glycans 540:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 3 46 theme substantial 518:528 arg1 N-glycans 560:568 such plant specific N-glycans 540:568 such plant specific N-glycans 540:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 3 46 theme substantial 518:528 arg1 amount 530:535 a substantial amount 516:535 a substantial amount of such plant specific N-glycans 516:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 1 47 theme pollen 202:207 arg1 Cry 219:221 Cry 219:221 Cry 219:221 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 47 theme pollen 202:207 arg1 allergen 209:216 The Japanese cedar pollen allergen 183:216 The Japanese cedar pollen allergen (Cry j1) 183:225 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 48 dep Lewis 334:338 arg1 s 350:350 a epitope(s) 340:351 Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-) 334:377 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 0 49 theme type 143:146 arg1 N-glycans 148:156 the plant complex type N-glycans 125:156 the plant complex type N-glycans bearing Lewis a epitope 125:180 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 1 50 theme pollen 250:255 arg1 allergen 257:264 the mountain cedar pollen allergen 231:264 the mountain cedar pollen allergen (Jun a1) 231:273 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 1 50 theme pollen 250:255 arg1 a1 271:272 Jun a1 267:272 Jun a1 267:272 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 5 51 theme a 1068:1068 arg1 type 1070:1073 the Lewis a type 1058:1073 the Lewis a type 1058:1073 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 2 52 theme biological 384:393 arg1 significance 395:406 The biological significance 380:406 The biological significance of Lewis a type plant N-glycans 380:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 3 53 theme epitope-containing 690:707 arg1 N-glycans 715:723 Lewis a epitope-containing plant N-glycans 682:723 Lewis a epitope-containing plant N-glycans 682:723 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 0 54 theme complex 135:141 arg1 N-glycans 148:156 the plant complex type N-glycans 125:156 the plant complex type N-glycans bearing Lewis a epitope 125:180 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 3 55 theme immunological 595:607 arg1 activity 609:616 immunological activity 595:616 immunological activity 595:616 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 5 56 theme complex 1025:1031 arg1 N-glycans 1038:1046 plant complex type N-glycans 1019:1046 plant complex type N-glycans 1019:1046 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 3 57 theme plant 709:713 arg1 N-glycans 715:723 Lewis a epitope-containing plant N-glycans 682:723 Lewis a epitope-containing plant N-glycans 682:723 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 3 58 theme such 540:543 arg1 N-glycans 560:568 such plant specific N-glycans 540:568 such plant specific N-glycans 540:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 5 59 theme all 943:945 arg1 glycoproteins 947:959 almost all glycoproteins 936:959 almost all glycoproteins expressed in these three water plants 936:997 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 60 theme Lewis 1147:1151 arg1 N-glycans 1166:1174 Lewis a type plant N-glycans 1147:1174 Lewis a type plant N-glycans 1147:1174 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 0 61 theme N-glycans 148:156 arg1 occurrence 111:120 Predominant occurrence 99:120 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 0:181 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 3 62 theme plant 545:549 arg1 N-glycans 560:568 such plant specific N-glycans 540:568 such plant specific N-glycans 540:568 Since a substantial amount of such plant specific N-glycans are required to evaluate immunological activity, we have searched for good plant-glycan sources to characterize Lewis a epitope-containing plant N-glycans. 27705835 4 63 gly glycoproteins 848:860 arg1 glycoproteins 848:860 glycoproteins 848:860 glycoproteins bearing Lewis a units 848:882 In this study, we have found that three water plants, Elodea nuttallii, Egeria densa, and Ceratophyllum demersum, produce glycoproteins bearing Lewis a units. 27705835 1 64 dep s 350:350 arg1 GlcNAc- 370:376 GlcNAc- 370:376 GlcNAc- 370:376 The Japanese cedar pollen allergen (Cry j1) and the mountain cedar pollen allergen (Jun a1) are glycosylated with plant complex type N-glycans bearing Lewis a epitope(s) (Galβ1-3[Fucα1-4]GlcNAc-). 27705835 2 65 theme N-glycans 430:438 arg1 significance 395:406 The biological significance 380:406 The biological significance of Lewis a type plant N-glycans 380:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 2 65 theme N-glycans 430:438 arg1 effects 450:456 their effects 444:456 their effects on the human immune system 444:483 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 2 66 from effects 450:456 arg1 system 478:483 the human immune system 461:483 the human immune system 461:483 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 0 67 gly glycoproteins 43:55 arg1 glycoproteins 43:55 glycoproteins 43:55 glycoproteins expressed in three kinds of water plants 43:96 Structural features of N-glycans linked to glycoproteins expressed in three kinds of water plants: Predominant occurrence of the plant complex type N-glycans bearing Lewis a epitope. 27705835 5 68 theme water 1098:1102 arg1 plants 1104:1109 these water plants 1092:1109 these water plants 1092:1109 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 68 theme water 1098:1102 arg1 sources 1120:1126 good sources 1115:1126 good sources for preparation of Lewis a type plant N-glycans in substantial amounts 1115:1197 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 69 theme type 1155:1158 arg1 N-glycans 1166:1174 Lewis a type plant N-glycans 1147:1174 Lewis a type plant N-glycans 1147:1174 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 2 70 theme plant 424:428 arg1 N-glycans 430:438 Lewis a type plant N-glycans 411:438 Lewis a type plant N-glycans 411:438 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27705835 5 71 theme a 1153:1153 arg1 N-glycans 1166:1174 Lewis a type plant N-glycans 1147:1174 Lewis a type plant N-glycans 1147:1174 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 5 72 theme N-glycans 1166:1174 arg1 preparation 1132:1142 preparation 1132:1142 preparation of Lewis a type plant N-glycans in substantial amounts 1132:1197 Structural analysis of the N-glycans revealed that almost all glycoproteins expressed in these three water plants predominantly carry plant complex type N-glycans including the Lewis a type, suggesting that these water plants are good sources for preparation of Lewis a type plant N-glycans in substantial amounts. 27705835 2 73 theme human 465:469 arg1 system 478:483 the human immune system 461:483 the human immune system 461:483 The biological significance of Lewis a type plant N-glycans and their effects on the human immune system remain to be elucidated. 27384922 5 0 theme specific 696:703 arg1 role 705:708 the specific role 692:708 the specific role of PrP(C) glycosylation status 692:739 Here, we investigated the specific role of PrP(C) glycosylation status. 27384922 7 1 theme strainness 1069:1078 arg1 perpetuation 1080:1091 strainness perpetuation 1069:1091 strainness perpetuation 1069:1091 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 2 2 theme host 315:318 arg1 C 324:324 C 324:324 C 324:324 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 27384922 2 2 theme host 315:318 arg1 PrP 320:322 host PrP 315:322 host PrP(C) 315:325 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 27384922 8 3 from backbone 1215:1222 arg1 glycans 1245:1251 the attached glycans 1232:1251 the attached glycans 1232:1251 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 3 4 theme glycotype 432:440 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 8 5 theme attached 1236:1243 arg1 glycans 1245:1251 the attached glycans 1232:1251 the attached glycans 1232:1251 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 8 6 theme PrP 1196:1198 arg1 backbone 1215:1222 PrP(Sc) structural backbone 1196:1222 PrP(Sc) structural backbone 1196:1222 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 8 7 theme structural 1204:1213 arg1 backbone 1215:1222 PrP(Sc) structural backbone 1196:1222 PrP(Sc) structural backbone 1196:1222 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 1 8 dep assemblies 144:153 arg1 PrP 156:158 PrP 156:158 PrP(Sc) 156:162 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 1 8 dep assemblies 144:153 arg1 Sc 160:161 Sc 160:161 Sc 160:161 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 1 9 gly N-glycosylated 181:194 arg1 protein 211:217 cellular prion protein 196:217 the variably N-glycosylated cellular prion protein (PrP(C)) 168:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 4 10 theme assemblies 545:554 arg1 conformation 521:532 the conformation 517:532 the conformation of PrP(Sc) assemblies 517:554 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 2 11 theme infected 232:239 arg1 species 241:247 infected species 232:247 infected species 232:247 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 27384922 7 12 theme glycoform 1003:1011 arg1 stoichiometry 1013:1025 glycoform stoichiometry 1003:1025 glycoform stoichiometry 1003:1025 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 6 13 theme amplification 801:813 arg1 method 815:820 an efficient protein misfolding cyclic amplification method 762:820 an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate 762:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 7 14 dep PrP 984:986 arg1 stoichiometry 1013:1025 glycoform stoichiometry 1003:1025 glycoform stoichiometry 1003:1025 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 3 15 theme Distinct 328:335 arg1 strains 343:349 Distinct prion strains 328:349 Distinct prion strains 328:349 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 3 16 theme defined 381:387 arg1 properties 409:418 defined PrP(Sc) biochemical properties 381:418 defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits 381:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 3 16 theme defined 381:387 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 0 17 theme prion 22:26 arg1 conversion 28:37 Glycoform-independent prion conversion 0:37 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 0:111 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 27384922 6 18 theme cyclic 794:799 arg1 method 815:820 an efficient protein misfolding cyclic amplification method 762:820 an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate 762:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 2 19 dep conversion 282:291 arg1 the 278:280 the 278:280 the 278:280 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 27384922 7 20 theme PrP 1047:1049 arg1 formation 1055:1063 PrP(Sc) formation 1047:1063 PrP(Sc) formation 1047:1063 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 4 21 theme information 587:597 arg1 storage 561:567 the storage 557:567 the storage of the structural information 557:597 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 4 21 theme information 587:597 arg1 requirements 617:628 the molecular requirements 603:628 the molecular requirements for self-perpetuation 603:650 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 0 22 theme Glycoform-independent 0:20 arg1 conversion 28:37 Glycoform-independent prion conversion 0:37 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 0:111 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 27384922 5 23 theme glycosylation 720:732 arg1 status 734:739 PrP(C) glycosylation status 713:739 PrP(C) glycosylation status 713:739 Here, we investigated the specific role of PrP(C) glycosylation status. 27384922 4 24 theme structural 576:585 arg1 information 587:597 the structural information 572:597 the structural information 572:597 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 5 25 theme status 734:739 arg1 role 705:708 the specific role 692:708 the specific role of PrP(C) glycosylation status 692:739 Here, we investigated the specific role of PrP(C) glycosylation status. 27384922 4 26 theme strain 480:485 arg1 information 487:497 strain information 480:497 strain information 480:497 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 8 27 theme strain 1127:1132 arg1 glycotype 1164:1172 PrP(Sc) glycotype 1156:1172 PrP(Sc) glycotype 1156:1172 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 8 27 theme strain 1127:1132 arg1 properties 1134:1143 strain properties 1127:1143 strain properties 1127:1143 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 6 28 theme interest 867:874 arg1 species 856:862 the PrP(C) species 845:862 the PrP(C) species of interest 845:874 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 6 28 theme interest 867:874 arg1 substrate 879:887 substrate 879:887 substrate 879:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 3 29 theme biological 455:464 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 6 30 theme misfolding 783:792 arg1 method 815:820 an efficient protein misfolding cyclic amplification method 762:820 an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate 762:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 4 31 theme molecular 607:615 arg1 requirements 617:628 the molecular requirements 603:628 the molecular requirements for self-perpetuation 603:650 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 1 32 theme N-glycosylated 181:194 arg1 protein 211:217 cellular prion protein 196:217 the variably N-glycosylated cellular prion protein (PrP(C)) 168:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 0 33 theme efficient 49:57 arg1 amplification 98:110 highly efficient, cell-based, protein misfolding cyclic amplification 42:110 highly efficient, cell-based, protein misfolding cyclic amplification 42:110 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 27384922 1 34 theme cellular 196:203 arg1 protein 211:217 cellular prion protein 196:217 the variably N-glycosylated cellular prion protein (PrP(C)) 168:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 3 35 theme PrP 389:391 arg1 properties 409:418 defined PrP(Sc) biochemical properties 381:418 defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits 381:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 3 35 theme PrP 389:391 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 5 36 theme PrP 713:715 arg1 status 734:739 PrP(C) glycosylation status 713:739 PrP(C) glycosylation status 713:739 Here, we investigated the specific role of PrP(C) glycosylation status. 27384922 1 37 dep protein 211:217 arg1 PrP 220:222 PrP 220:222 PrP(C) 220:225 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 1 37 dep protein 211:217 arg1 C 224:224 C 224:224 C 224:224 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 1 38 theme prion 205:209 arg1 protein 211:217 cellular prion protein 196:217 the variably N-glycosylated cellular prion protein (PrP(C)) 168:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 3 39 theme prion 337:341 arg1 strains 343:349 Distinct prion strains 328:349 Distinct prion strains 328:349 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 0 40 dep efficient 49:57 arg1 cyclic 91:96 cyclic 91:96 cyclic 91:96 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 27384922 0 40 dep efficient 49:57 arg1 cell-based 60:69 cell-based 60:69 cell-based 60:69 Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification. 27384922 8 41 theme PrP 1156:1158 arg1 glycotype 1164:1172 PrP(Sc) glycotype 1156:1172 PrP(Sc) glycotype 1156:1172 Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans. 27384922 4 42 theme PrP 537:539 arg1 assemblies 545:554 PrP(Sc) assemblies 537:554 PrP(Sc) assemblies 537:554 While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. 27384922 6 43 theme efficient 765:773 arg1 method 815:820 an efficient protein misfolding cyclic amplification method 762:820 an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate 762:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 7 44 theme PrP 916:918 arg1 mutants 937:943 PrP(C) glycosylation mutants 916:943 PrP(C) glycosylation mutants expressing cells 916:960 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 1 45 theme misfolded 134:142 arg1 assemblies 144:153 misfolded assemblies 134:153 misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)) 134:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 1 46 theme protein 211:217 arg1 assemblies 144:153 misfolded assemblies 134:153 misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)) 134:226 Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). 27384922 7 47 theme glycosylation 923:935 arg1 mutants 937:943 PrP(C) glycosylation mutants 916:943 PrP(C) glycosylation mutants expressing cells 916:960 Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. 27384922 3 48 theme specific 446:453 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 3 49 theme biochemical 397:407 arg1 properties 409:418 defined PrP(Sc) biochemical properties 381:418 defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits 381:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 3 49 theme biochemical 397:407 arg1 traits 466:471 the glycotype and specific biological traits 428:471 the glycotype and specific biological traits 428:471 Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. 27384922 6 50 theme protein 775:781 arg1 method 815:820 an efficient protein misfolding cyclic amplification method 762:820 an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate 762:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 6 51 theme PrP 849:851 arg1 species 856:862 the PrP(C) species 845:862 the PrP(C) species of interest 845:874 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 6 51 theme PrP 849:851 arg1 substrate 879:887 substrate 879:887 substrate 879:887 First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. 27384922 2 52 theme PrP 320:322 arg1 polymerization 297:310 polymerization 297:310 polymerization 297:310 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 27384922 2 52 theme PrP 320:322 arg1 conversion 282:291 conversion 282:291 conversion 282:291 In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). 26797104 6 0 theme nDer 1069:1072 arg1 p 1074:1074 nDer p 23 1069:1077 nDer p 23 1069:1077 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 6 0 theme nDer 1069:1072 arg1 allergen 1059:1066 the corresponding natural allergen 1033:1066 the corresponding natural allergen (nDer p 23) 1033:1078 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 4 1 theme degranulation 849:861 arg1 assays 863:868 RBL-SX38 degranulation assays 840:868 RBL-SX38 degranulation assays 840:868 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 3 2 theme dichroism 692:700 arg1 analyses 702:709 circular dichroism analyses 683:709 circular dichroism analyses as well as for its chitin-binding activity 683:752 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 1 3 theme identified 180:189 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 11 4 theme component-resolved 1702:1719 arg1 diagnosis 1721:1729 HDM allergy component-resolved diagnosis 1690:1729 HDM allergy component-resolved diagnosis than HDM extracts 1690:1747 Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts. 26797104 1 5 theme dust 197:200 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 5 6 contain carried 919:925 arg2 O-mannosylation 927:941 O-mannosylation 927:941 O-mannosylation 927:941 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 5 6 contain carried 919:925 arg1 RESULTS 885:891 RESULTS 885:891 RESULTS Purified intact rDer p 23 885:917 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 11 7 theme HDM 1690:1692 arg1 diagnosis 1721:1729 HDM allergy component-resolved diagnosis 1690:1729 HDM allergy component-resolved diagnosis than HDM extracts 1690:1747 Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts. 26797104 11 8 theme HDM 1736:1738 arg1 extracts 1740:1747 HDM extracts 1736:1747 HDM extracts 1736:1747 Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts. 26797104 2 9 theme patients 545:552 arg1 cohort 522:527 a cohort 520:527 a cohort of HDM-allergic patients from Thailand 520:566 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 1 10 dep allergen 219:226 arg1 p 232:232 Der p 23 228:235 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 6 11 theme B-cell 1166:1171 arg1 epitopes 1173:1180 common B-cell epitopes 1159:1180 common B-cell epitopes 1159:1180 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 5 12 theme Purified 893:900 arg1 rDer 909:912 intact rDer 902:912 RESULTS Purified intact rDer p 23 885:917 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 8 13 theme 23-specific 1389:1399 arg1 responses 1405:1413 Der p 23-specific IgE responses 1383:1413 Der p 23-specific IgE responses 1383:1413 Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. 26797104 6 14 theme p 1148:1148 arg1 forms 1135:1139 both forms 1130:1139 both forms of Der p 23 1130:1151 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 11 15 theme rDer 1649:1652 arg1 p 1654:1654 rDer p 23 1649:1657 rDer p 23 1649:1657 Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts. 26797104 3 16 theme rDer 586:589 arg1 p 591:591 Purified rDer p 23 577:594 METHODS Purified rDer p 23 569:594 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 6 17 theme Polyclonal 987:996 arg1 antibodies 998:1007 Polyclonal antibodies 987:1007 Polyclonal antibodies to rDer p 23 987:1020 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 0 18 theme O-Glycan-Independent 53:72 arg1 Reactivities 78:89 O-Glycan-Independent IgE Reactivities 53:89 O-Glycan-Independent IgE Reactivities 53:89 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 9 19 theme rDer 1446:1449 arg1 p 1451:1451 rDer p 23 1446:1454 rDer p 23 1446:1454 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 1 20 dep BACKGROUND 123:132 arg1 requires 237:244 requires 237:244 requires the production of its recombinant counterpart because the natural allergen 237:319 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 1 21 theme fecal 348:352 arg1 pellets 354:360 fecal pellets 348:360 fecal pellets 348:360 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 5 22 dep RESULTS 885:891 arg1 rDer 909:912 intact rDer 902:912 RESULTS Purified intact rDer p 23 885:917 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 6 23 theme natural 1051:1057 arg1 p 1074:1074 nDer p 23 1069:1077 nDer p 23 1069:1077 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 6 23 theme natural 1051:1057 arg1 allergen 1059:1066 the corresponding natural allergen 1033:1066 the corresponding natural allergen (nDer p 23) 1033:1078 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 2 24 theme p 455:455 arg1 characterization 419:434 a detailed physico-chemical characterization 391:434 a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) 391:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 10 25 theme important 1559:1567 arg1 levels 1569:1574 important levels 1559:1574 important levels of Der p 23 sensitizations in Thailand 1559:1613 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 7 26 theme rDer 1262:1265 arg1 p 1267:1267 rDer p 23 1262:1270 rDer p 23 1262:1270 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 2 27 theme recombinant 439:449 arg1 p 455:455 recombinant Der p 23 439:458 recombinant Der p 23 (rDer p 23) 439:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 2 27 theme recombinant 439:449 arg1 p 466:466 rDer p 23 461:469 rDer p 23 461:469 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 10 28 theme sensitizations 1588:1601 arg1 levels 1569:1574 important levels 1559:1574 important levels of Der p 23 sensitizations in Thailand 1559:1613 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 0 29 theme Dust 10:13 arg1 Allergen 26:33 The House Dust Mite Major Allergen 0:33 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 1 30 theme recombinant 268:278 arg1 counterpart 280:290 its recombinant counterpart 264:290 its recombinant counterpart 264:290 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 1 31 from pellets 354:360 arg1 extractable 331:341 extractable 331:341 extractable 331:341 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 3 32 theme recombinant 611:621 arg1 pastoris 630:637 recombinant Pichia pastoris 611:637 recombinant Pichia pastoris 611:637 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 2 33 theme detailed 393:400 arg1 characterization 419:434 a detailed physico-chemical characterization 391:434 a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) 391:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 0 34 theme Major 20:24 arg1 Allergen 26:33 The House Dust Mite Major Allergen 0:33 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 9 35 theme basophil 1498:1505 arg1 cells 1516:1520 rat basophil leukemia cells 1494:1520 rat basophil leukemia cells 1494:1520 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 7 36 theme Der 1249:1251 arg1 p 1253:1253 both natural Der p 23 1236:1256 both natural Der p 23 1236:1256 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 4 37 theme p 806:806 arg1 frequency 771:779 IgE-binding frequency 759:779 IgE-binding frequency 759:779 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 4 37 theme p 806:806 arg1 allergenicity 785:797 allergenicity 785:797 allergenicity 785:797 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 0 38 theme Der 35:37 arg1 p 39:39 Der p 23 35:42 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 1 39 theme natural 304:310 arg1 allergen 312:319 the natural allergen 300:319 the natural allergen 300:319 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 9 40 theme cells 1516:1520 arg1 degranulation 1477:1489 the degranulation 1473:1489 the degranulation of rat basophil leukemia cells 1473:1520 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 6 41 theme aqueous 1083:1089 arg1 extracts 1104:1111 aqueous fecal pellet extracts 1083:1111 aqueous fecal pellet extracts 1083:1111 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 8 42 theme HDM-allergic 1339:1350 arg1 patients 1352:1359 222 Thai HDM-allergic patients 1330:1359 222 Thai HDM-allergic patients tested 1330:1366 Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. 26797104 1 43 theme major 213:217 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 5 44 theme coil 971:974 arg1 structure 976:984 a random coil structure 962:984 a random coil structure 962:984 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 4 45 dep frequency 771:779 arg1 The 755:757 The 755:757 The 755:757 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 1 46 theme Der 228:230 arg1 p 232:232 Der p 23 228:235 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 10 47 dep CONCLUSION 1523:1532 arg1 highlighted 1547:1557 highlighted 1547:1557 highlighted important levels of Der p 23 sensitizations in Thailand 1547:1613 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 2 48 from cohort 522:527 arg1 Thailand 559:566 Thailand 559:566 Thailand 559:566 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 7 49 with homologies 1195:1204 arg1 proteins 1226:1233 chitin-binding proteins 1211:1233 chitin-binding proteins 1211:1233 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 8 50 theme 222 1330:1332 arg1 patients 1352:1359 222 Thai HDM-allergic patients 1330:1359 222 Thai HDM-allergic patients tested 1330:1366 Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. 26797104 4 51 theme RBL-SX38 840:847 arg1 assays 863:868 RBL-SX38 degranulation assays 840:868 RBL-SX38 degranulation assays 840:868 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 10 52 theme p 1583:1583 arg1 sensitizations 1588:1601 Der p 23 sensitizations 1579:1601 Der p 23 sensitizations 1579:1601 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 3 53 theme circular 683:690 arg1 dichroism 692:700 circular dichroism 683:700 circular dichroism analyses as well as for its chitin-binding activity 683:752 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 1 54 theme house 191:195 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 1 55 theme mite 202:205 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 6 56 theme pellet 1097:1102 arg1 extracts 1104:1111 aqueous fecal pellet extracts 1083:1111 aqueous fecal pellet extracts 1083:1111 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 11 57 theme allergy 1694:1700 arg1 diagnosis 1721:1729 HDM allergy component-resolved diagnosis 1690:1729 HDM allergy component-resolved diagnosis than HDM extracts 1690:1747 Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts. 26797104 3 58 dep METHODS 569:575 arg1 p 591:591 Purified rDer p 23 577:594 METHODS Purified rDer p 23 569:594 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 1 59 theme HDM 208:210 arg1 allergen 219:226 the recently identified house dust mite (HDM) major allergen 167:226 the recently identified house dust mite (HDM) major allergen Der p 23 167:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 2 60 theme IgE 502:504 arg1 reactivity 506:515 its IgE reactivity 498:515 its IgE reactivity 498:515 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 7 61 theme chitin 1310:1315 arg1 matrices 1317:1324 chitin matrices 1310:1324 chitin matrices 1310:1324 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 2 62 theme HDM-allergic 532:543 arg1 patients 545:552 HDM-allergic patients 532:552 HDM-allergic patients from Thailand 532:566 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 6 63 theme common 1159:1164 arg1 epitopes 1173:1180 common B-cell epitopes 1159:1180 common B-cell epitopes 1159:1180 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 3 64 theme chitin-binding 730:743 arg1 activity 745:752 its chitin-binding activity 726:752 its chitin-binding activity 726:752 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 4 65 theme IgE-binding 759:769 arg1 frequency 771:779 IgE-binding frequency 759:779 IgE-binding frequency 759:779 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 5 66 theme intact 902:907 arg1 rDer 909:912 intact rDer 902:912 RESULTS Purified intact rDer p 23 885:917 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 6 67 theme rDer 1012:1015 arg1 p 1017:1017 rDer p 23 1012:1020 rDer p 23 1012:1020 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 8 68 theme IgE 1401:1403 arg1 responses 1405:1413 Der p 23-specific IgE responses 1383:1413 Der p 23-specific IgE responses 1383:1413 Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. 26797104 3 69 theme Purified 577:584 arg1 p 591:591 Purified rDer p 23 577:594 METHODS Purified rDer p 23 569:594 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 1 70 theme in-depth 138:145 arg1 characterization 147:162 The in-depth characterization 134:162 The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 134:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 6 71 theme Der 1144:1146 arg1 p 1148:1148 Der p 23 1144:1151 Der p 23 1144:1151 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 0 72 theme IgE 74:76 arg1 Reactivities 78:89 O-Glycan-Independent IgE Reactivities 53:89 O-Glycan-Independent IgE Reactivities 53:89 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 9 73 theme p 1451:1451 arg1 allergenicity 1429:1441 the allergenicity 1425:1441 the allergenicity of rDer p 23 1425:1454 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 5 74 dep rDer 909:912 arg1 p 914:914 p 23 914:917 RESULTS Purified intact rDer p 23 885:917 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 0 75 theme Chitin-Binding 98:111 arg1 Activity 113:120 No Chitin-Binding Activity 95:120 No Chitin-Binding Activity 95:120 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 0 76 theme House 4:8 arg1 Allergen 26:33 The House Dust Mite Major Allergen 0:33 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 6 77 theme corresponding 1037:1049 arg1 p 1074:1074 nDer p 23 1069:1077 nDer p 23 1069:1077 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 6 77 theme corresponding 1037:1049 arg1 allergen 1059:1066 the corresponding natural allergen 1033:1066 the corresponding natural allergen (nDer p 23) 1033:1078 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 2 78 theme Der 451:453 arg1 p 455:455 recombinant Der p 23 439:458 recombinant Der p 23 (rDer p 23) 439:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 2 78 theme Der 451:453 arg1 p 466:466 rDer p 23 461:469 rDer p 23 461:469 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 2 79 from Thailand 559:566 arg1 cohort 522:527 a cohort 520:527 a cohort of HDM-allergic patients from Thailand 520:566 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 2 79 from Thailand 559:566 arg1 patients 545:552 HDM-allergic patients 532:552 HDM-allergic patients from Thailand 532:566 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 0 80 theme Mite 15:18 arg1 Allergen 26:33 The House Dust Mite Major Allergen 0:33 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 2 81 theme physico-chemical 402:417 arg1 characterization 419:434 a detailed physico-chemical characterization 391:434 a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) 391:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 1 82 theme counterpart 280:290 arg1 production 250:259 the production 246:259 the production of its recombinant counterpart 246:290 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 9 83 theme rat 1494:1496 arg1 cells 1516:1520 rat basophil leukemia cells 1494:1520 rat basophil leukemia cells 1494:1520 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 7 84 theme natural 1241:1247 arg1 p 1253:1253 both natural Der p 23 1236:1256 both natural Der p 23 1236:1256 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 2 85 theme rDer 461:464 arg1 p 455:455 recombinant Der p 23 439:458 recombinant Der p 23 (rDer p 23) 439:470 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 2 85 theme rDer 461:464 arg1 p 466:466 rDer p 23 461:469 rDer p 23 461:469 This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. 26797104 10 86 theme Der 1579:1581 arg1 sensitizations 1588:1601 Der p 23 sensitizations 1579:1601 Der p 23 sensitizations 1579:1601 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 0 87 dep Allergen 26:33 arg1 p 39:39 Der p 23 35:42 The House Dust Mite Major Allergen Der p 23 0:42 The House Dust Mite Major Allergen Der p 23 Displays O-Glycan-Independent IgE Reactivities but No Chitin-Binding Activity. 26797104 6 88 theme fecal 1091:1095 arg1 extracts 1104:1111 aqueous fecal pellet extracts 1083:1111 aqueous fecal pellet extracts 1083:1111 Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. 26797104 4 89 theme Der 802:804 arg1 p 806:806 Der p 23 802:809 Der p 23 802:809 The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. 26797104 9 90 theme leukemia 1507:1514 arg1 cells 1516:1520 rat basophil leukemia cells 1494:1520 rat basophil leukemia cells 1494:1520 Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. 26797104 5 91 theme random 964:969 arg1 structure 976:984 a random coil structure 962:984 a random coil structure 962:984 RESULTS Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. 26797104 10 92 from levels 1569:1574 arg1 Thailand 1606:1613 Thailand 1606:1613 Thailand 1606:1613 CONCLUSION Our findings highlighted important levels of Der p 23 sensitizations in Thailand. 26797104 7 93 theme chitin-binding 1211:1224 arg1 proteins 1226:1233 chitin-binding proteins 1211:1233 chitin-binding proteins 1211:1233 Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. 26797104 3 94 theme Pichia 623:628 arg1 pastoris 630:637 recombinant Pichia pastoris 611:637 recombinant Pichia pastoris 611:637 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 26797104 8 95 theme Thai 1334:1337 arg1 patients 1352:1359 222 Thai HDM-allergic patients 1330:1359 222 Thai HDM-allergic patients tested 1330:1366 Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. 26797104 1 96 theme allergen 219:226 arg1 characterization 147:162 The in-depth characterization 134:162 The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 134:235 BACKGROUND The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. 26797104 3 97 theme mass 661:664 arg1 spectrometry 666:677 mass spectrometry 661:677 mass spectrometry 661:677 METHODS Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. 29017882 0 0 theme Cassia 94:99 arg1 seeds 113:117 Cassia obtusifolia seeds 94:117 Cassia obtusifolia seeds 94:117 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 4 1 theme CP-40-M 783:789 arg1 glucuronoxylan 812:825 glucuronoxylan 812:825 glucuronoxylan 812:825 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 4 1 theme CP-40-M 783:789 arg1 structure 770:778 The structure 766:778 The structure of CP-40-M 766:789 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 4 2 with glucuronoxylan 812:825 arg1 group 859:863 glucopyranosyluronic acid group 833:863 glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone 833:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 4 3 attach attached 876:883 arg2 group 859:863 glucopyranosyluronic acid group 833:863 glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone 833:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 4 3 attach attached 876:883 arg1 O-2 888:890 O-2 888:890 O-2 of the →4)-β-Xylp-(1→ backbone 888:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 1 4 theme homogeneous 256:266 arg1 CP-40-M 283:289 a homogeneous polysaccharide CP-40-M 254:289 a homogeneous polysaccharide CP-40-M 254:289 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 0 5 theme seeds 113:117 arg1 extract 83:89 water extract 77:89 water extract of Cassia obtusifolia seeds 77:117 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 3 6 theme molar 715:719 arg1 ratio 721:725 the molar ratio 711:725 the molar ratio of xylose to glucuronic acid 711:754 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 3 6 theme molar 715:719 arg1 4.62 760:763 4.62 760:763 4.62 760:763 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 4 7 theme →4 899:900 arg1 backbone 914:921 the →4)-β-Xylp-(1→ backbone 895:921 the →4)-β-Xylp-(1→ backbone 895:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 5 8 theme Cassia 1001:1006 arg1 seeds 1020:1024 Cassia obtusifolia seeds 1001:1024 Cassia obtusifolia seeds 1001:1024 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 6 9 theme cassia 1135:1140 arg1 polysaccharide 1142:1155 cassia polysaccharide 1135:1155 cassia polysaccharide 1135:1155 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 1 10 theme polysaccharide 268:281 arg1 CP-40-M 283:289 a homogeneous polysaccharide CP-40-M 254:289 a homogeneous polysaccharide CP-40-M 254:289 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 0 11 theme obtusifolia 101:111 arg1 seeds 113:117 Cassia obtusifolia seeds 94:117 Cassia obtusifolia seeds 94:117 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 3 12 theme glucuronic 740:749 arg1 acid 751:754 glucuronic acid 740:754 glucuronic acid 740:754 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 6 13 theme potential 1181:1189 arg1 use 1191:1193 their potential use 1175:1193 their potential use in food industry and folk medicine 1175:1228 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 2 14 theme multiple 436:443 arg1 SEC-MALLS 456:464 SEC-MALLS 456:464 SEC-MALLS 456:464 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 14 theme multiple 436:443 arg1 detectors 445:453 multiple detectors 436:453 multiple detectors (SEC-MALLS) 436:465 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 3 15 theme xylose 730:735 arg1 ratio 721:725 the molar ratio 711:725 the molar ratio of xylose to glucuronic acid 711:754 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 3 15 theme xylose 730:735 arg1 4.62 760:763 4.62 760:763 4.62 760:763 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 2 16 theme anion-exchange 485:498 arg1 HPAEC 516:520 HPAEC 516:520 HPAEC 516:520 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 16 theme anion-exchange 485:498 arg1 chromatography 407:420 size-exclusion chromatography 392:420 size-exclusion chromatography equipped with multiple detectors (SEC-MALLS) 392:465 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 16 theme anion-exchange 485:498 arg1 chromatography 500:513 high-performance anion-exchange chromatography 468:513 high-performance anion-exchange chromatography (HPAEC) 468:521 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 6 17 from use 1191:1193 arg1 industry 1203:1210 food industry 1198:1210 food industry 1198:1210 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 6 17 from use 1191:1193 arg1 medicine 1221:1228 folk medicine 1216:1228 folk medicine 1216:1228 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 6 18 theme CP-40-M 1056:1062 arg1 meaningful 1068:1077 meaningful 1068:1077 meaningful 1068:1077 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 6 18 theme CP-40-M 1056:1062 arg1 elucidation 1041:1051 The structure elucidation 1027:1051 The structure elucidation of CP-40-M 1027:1062 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 0 19 theme Enzymatic 0:8 arg1 purification 10:21 Enzymatic purification 0:21 Enzymatic purification 0:21 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 5 20 theme first 935:939 arg1 time 941:944 the first time 931:944 the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds 931:1024 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 5 20 theme first 935:939 arg1 It 924:925 It 924:925 It 924:925 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 0 21 theme structure 27:35 arg1 characterization 37:52 structure characterization 27:52 structure characterization 27:52 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 6 22 theme food 1198:1201 arg1 industry 1203:1210 food industry 1198:1210 food industry 1198:1210 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 2 23 theme nuclear 584:590 arg1 resonance 601:609 nuclear magnetic resonance 584:609 nuclear magnetic resonance (NMR) spectra 584:623 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 23 theme nuclear 584:590 arg1 NMR 612:614 NMR 612:614 NMR 612:614 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 5 24 theme obtusifolia 1008:1018 arg1 seeds 1020:1024 Cassia obtusifolia seeds 1001:1024 Cassia obtusifolia seeds 1001:1024 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 6 25 theme polysaccharide 1142:1155 arg1 characteristics 1116:1130 the natural characteristics 1104:1130 the natural characteristics of cassia polysaccharide 1104:1155 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 1 26 theme water-extracted 203:217 arg1 CP-40 242:246 CP-40 242:246 CP-40 242:246 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 1 26 theme water-extracted 203:217 arg1 polysaccharide 226:239 water-extracted cassia polysaccharide 203:239 water-extracted cassia polysaccharide (CP-40) 203:247 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 4 27 theme backbone 914:921 arg1 O-2 888:890 O-2 888:890 O-2 of the →4)-β-Xylp-(1→ backbone 888:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 1 28 theme Enzymatic 120:128 arg1 hydrolysis 130:139 Enzymatic hydrolysis 120:139 Enzymatic hydrolysis 120:139 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 1 29 theme cassia 219:224 arg1 CP-40 242:246 CP-40 242:246 CP-40 242:246 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 1 29 theme cassia 219:224 arg1 polysaccharide 226:239 water-extracted cassia polysaccharide 203:239 water-extracted cassia polysaccharide (CP-40) 203:247 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 1 30 theme polysaccharide 226:239 arg1 sub-fraction 187:198 the sub-fraction 183:198 the sub-fraction of water-extracted cassia polysaccharide (CP-40) 183:247 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 1 31 theme high 320:323 arg1 yield 325:329 relatively high yield 309:329 relatively high yield 309:329 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 29017882 0 32 from extract 83:89 arg1 purification 10:21 Enzymatic purification 0:21 Enzymatic purification 0:21 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 0 32 from extract 83:89 arg1 characterization 37:52 structure characterization 27:52 structure characterization 27:52 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 2 33 theme high-performance 468:483 arg1 HPAEC 516:520 HPAEC 516:520 HPAEC 516:520 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 33 theme high-performance 468:483 arg1 chromatography 407:420 size-exclusion chromatography 392:420 size-exclusion chromatography equipped with multiple detectors (SEC-MALLS) 392:465 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 33 theme high-performance 468:483 arg1 chromatography 500:513 high-performance anion-exchange chromatography 468:513 high-performance anion-exchange chromatography (HPAEC) 468:521 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 4 34 theme -β-Xylp- 902:909 arg1 backbone 914:921 the →4)-β-Xylp-(1→ backbone 895:921 the →4)-β-Xylp-(1→ backbone 895:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 0 35 theme water 77:81 arg1 extract 83:89 water extract 77:89 water extract of Cassia obtusifolia seeds 77:117 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 4 36 theme acid 854:857 arg1 group 859:863 glucopyranosyluronic acid group 833:863 glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone 833:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 4 37 theme 1→ 911:912 arg1 backbone 914:921 the →4)-β-Xylp-(1→ backbone 895:921 the →4)-β-Xylp-(1→ backbone 895:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 2 38 theme CP-40-M 359:365 arg1 features 347:354 The structural features 332:354 The structural features of CP-40-M 332:365 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 0 39 theme glucuronoxylan 57:70 arg1 purification 10:21 Enzymatic purification 0:21 Enzymatic purification 0:21 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 0 39 theme glucuronoxylan 57:70 arg1 characterization 37:52 structure characterization 27:52 structure characterization 27:52 Enzymatic purification and structure characterization of glucuronoxylan from water extract of Cassia obtusifolia seeds. 29017882 5 40 theme seeds 1020:1024 arg1 extract 990:996 the water extract 980:996 the water extract of Cassia obtusifolia seeds 980:1024 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 4 41 theme glucopyranosyluronic 833:852 arg1 group 859:863 glucopyranosyluronic acid group 833:863 glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone 833:921 The structure of CP-40-M was elucidated to be glucuronoxylan, with glucopyranosyluronic acid group terminally attached to O-2 of the →4)-β-Xylp-(1→ backbone. 29017882 6 42 theme folk 1216:1219 arg1 medicine 1221:1228 folk medicine 1216:1228 folk medicine 1216:1228 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 5 43 theme water 984:988 arg1 extract 990:996 the water extract 980:996 the water extract of Cassia obtusifolia seeds 980:1024 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 2 44 theme resonance 601:609 arg1 spectra 617:623 nuclear magnetic resonance (NMR) spectra 584:623 nuclear magnetic resonance (NMR) spectra 584:623 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 5 45 theme xylan 969:973 arg1 type 961:964 this type 956:964 this type of xylan 956:973 It was the first time to obtain this type of xylan from the water extract of Cassia obtusifolia seeds. 29017882 6 46 theme natural 1108:1114 arg1 characteristics 1116:1130 the natural characteristics 1104:1130 the natural characteristics of cassia polysaccharide 1104:1155 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 2 47 theme magnetic 592:599 arg1 resonance 601:609 nuclear magnetic resonance 584:609 nuclear magnetic resonance (NMR) spectra 584:623 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 47 theme magnetic 592:599 arg1 NMR 612:614 NMR 612:614 NMR 612:614 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 48 theme structural 336:345 arg1 features 347:354 The structural features 332:354 The structural features of CP-40-M 332:365 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 3 49 theme weight-average 630:643 arg1 0.29×105Da 695:704 0.29×105Da 695:704 0.29×105Da 695:704 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 3 49 theme weight-average 630:643 arg1 weight 655:660 The weight-average molecular weight 626:660 The weight-average molecular weight for CP-40-M 626:672 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 6 50 theme structure 1031:1039 arg1 meaningful 1068:1077 meaningful 1068:1077 meaningful 1068:1077 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 6 50 theme structure 1031:1039 arg1 elucidation 1041:1051 The structure elucidation 1027:1051 The structure elucidation of CP-40-M 1027:1062 The structure elucidation of CP-40-M was meaningful for better understanding the natural characteristics of cassia polysaccharide and important for their potential use in food industry and folk medicine. 29017882 2 51 theme size-exclusion 392:405 arg1 chromatography 407:420 size-exclusion chromatography 392:420 size-exclusion chromatography equipped with multiple detectors (SEC-MALLS) 392:465 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 51 theme size-exclusion 392:405 arg1 chromatography 500:513 high-performance anion-exchange chromatography 468:513 high-performance anion-exchange chromatography (HPAEC) 468:521 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 51 theme size-exclusion 392:405 arg1 methylation 524:534 methylation 524:534 methylation 524:534 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 51 theme size-exclusion 392:405 arg1 chromatography-mass 544:562 gas chromatography-mass 540:562 gas chromatography-mass (GC-MS) 540:570 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 52 theme gas 540:542 arg1 GC-MS 565:569 GC-MS 565:569 GC-MS 565:569 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 52 theme gas 540:542 arg1 chromatography 407:420 size-exclusion chromatography 392:420 size-exclusion chromatography equipped with multiple detectors (SEC-MALLS) 392:465 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 2 52 theme gas 540:542 arg1 chromatography-mass 544:562 gas chromatography-mass 540:562 gas chromatography-mass (GC-MS) 540:570 The structural features of CP-40-M were characterized using size-exclusion chromatography equipped with multiple detectors (SEC-MALLS), high-performance anion-exchange chromatography (HPAEC), methylation and gas chromatography-mass (GC-MS), as well as nuclear magnetic resonance (NMR) spectra. 29017882 3 53 theme molecular 645:653 arg1 0.29×105Da 695:704 0.29×105Da 695:704 0.29×105Da 695:704 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 3 53 theme molecular 645:653 arg1 weight 655:660 The weight-average molecular weight 626:660 The weight-average molecular weight for CP-40-M 626:672 The weight-average molecular weight for CP-40-M was determined to be 0.29×105Da, and the molar ratio of xylose to glucuronic acid was 4.62. 29017882 1 54 theme first 162:166 arg1 time 168:171 the first time 158:171 the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40) 158:247 Enzymatic hydrolysis was proposed for the first time to purify the sub-fraction of water-extracted cassia polysaccharide (CP-40), and a homogeneous polysaccharide CP-40-M was obtained with relatively high yield. 27547921 5 0 theme insulin 735:741 arg1 stimulation 743:753 insulin stimulation 735:753 insulin stimulation 735:753 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 5 1 from retention 792:800 arg1 membrane 814:821 the cell membrane 805:821 the cell membrane 805:821 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 7 2 theme imaging 1009:1015 arg1 tool 1017:1020 this imaging tool 1004:1020 this imaging tool 1004:1020 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 4 3 theme exocytosed 568:577 arg1 GLUT4 579:583 exocytosed GLUT4 568:583 exocytosed GLUT4 568:583 One of the probes, which is cell impermeant, selectively detected exocytosed GLUT4. 27547921 7 4 theme dynamic 1093:1099 arg1 changes 1101:1107 dynamic changes 1093:1107 dynamic changes in localization 1093:1123 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 2 5 theme GLUT4 276:280 arg1 N-glycan 264:271 the N-glycan 260:271 the N-glycan of GLUT4 260:280 To date, it has been unclear whether the N-glycan of GLUT4 contributes to its intracellular trafficking. 27547921 2 6 theme intracellular 301:313 arg1 trafficking 315:325 its intracellular trafficking 297:325 its intracellular trafficking 297:325 To date, it has been unclear whether the N-glycan of GLUT4 contributes to its intracellular trafficking. 27547921 7 7 theme various 1057:1063 arg1 proteins 1074:1081 various membrane proteins 1057:1081 various membrane proteins that show dynamic changes in localization 1057:1123 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 3 8 theme multicolor 459:468 arg1 imaging 470:476 multicolor imaging 459:476 multicolor imaging of GLUT4 translocation 459:499 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 1 9 theme glucose 162:168 arg1 homeostasis 170:180 glucose homeostasis 162:180 glucose homeostasis 162:180 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 5 10 theme cell 716:719 arg1 membrane 721:728 the cell membrane 712:728 the cell membrane 712:728 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 7 11 theme membrane 1065:1072 arg1 proteins 1074:1081 various membrane proteins 1057:1081 various membrane proteins that show dynamic changes in localization 1057:1123 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 3 12 theme N-glycan 361:368 arg1 role 349:352 the role 345:352 the role of the N-glycan 345:368 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 0 13 from role 28:31 arg1 trafficking 75:85 intracellular trafficking 61:85 intracellular trafficking 61:85 Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking. 27547921 6 14 from retention 888:896 arg1 membrane 919:926 the cell membrane 910:926 the cell membrane 910:926 The results strongly suggest that the N-glycan functions in the retention of GLUT4 on the cell membrane. 27547921 1 15 theme Glucose 88:94 arg1 transporter 96:106 Glucose transporter 4 88:108 Glucose transporter 4 (GLUT4) 88:116 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 1 15 theme Glucose 88:94 arg1 GLUT4 111:115 GLUT4 111:115 GLUT4 111:115 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 1 15 theme Glucose 88:94 arg1 protein 139:145 an N-glycosylated protein 121:145 an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation 121:220 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 0 16 theme Fluorogenic 0:10 arg1 probes 12:17 Fluorogenic probes 0:17 Fluorogenic probes 0:17 Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking. 27547921 7 17 from research 1045:1052 arg1 proteins 1074:1081 various membrane proteins 1057:1081 various membrane proteins that show dynamic changes in localization 1057:1123 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 4 18 theme cell 530:533 arg1 impermeant 535:544 cell impermeant 530:544 cell impermeant 530:544 One of the probes, which is cell impermeant, selectively detected exocytosed GLUT4. 27547921 5 19 theme N-glycan-deficient 655:672 arg1 GLUT4 674:678 N-glycan-deficient GLUT4 655:678 N-glycan-deficient GLUT4 655:678 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 5 20 theme cell 809:812 arg1 membrane 814:821 the cell membrane 805:821 the cell membrane 805:821 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 1 21 theme protein 200:206 arg1 translocation 208:220 the protein translocation 196:220 the protein translocation 196:220 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 3 22 theme fluorogenic 384:394 arg1 probes 396:401 fluorogenic probes 384:401 fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation 384:499 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 7 23 from changes 1101:1107 arg1 localization 1112:1123 localization 1112:1123 localization 1112:1123 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 0 24 theme N-glycosylation 42:56 arg1 role 28:31 a role 26:31 a role of GLUT4 N-glycosylation in intracellular trafficking 26:85 Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking. 27547921 4 25 theme probes 513:518 arg1 One 502:504 One 502:504 One 502:504 One of the probes, which is cell impermeant, selectively detected exocytosed GLUT4. 27547921 4 25 theme probes 513:518 arg1 probes 513:518 the probes 509:518 the probes 509:518 One of the probes, which is cell impermeant, selectively detected exocytosed GLUT4. 27547921 5 26 theme trafficking 633:643 arg1 log 621:623 the 'log' 616:624 the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane 616:821 Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. 27547921 3 27 dep cytoplasmic 414:424 arg1 proteins 446:453 proteins 446:453 proteins 446:453 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 3 28 theme GLUT4 481:485 arg1 translocation 487:499 GLUT4 translocation 481:499 GLUT4 translocation 481:499 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 0 29 theme GLUT4 36:40 arg1 N-glycosylation 42:56 GLUT4 N-glycosylation 36:56 GLUT4 N-glycosylation 36:56 Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking. 27547921 1 30 gly N-glycosylated 124:137 arg1 protein 139:145 an N-glycosylated protein 121:145 an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation 121:220 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 1 30 gly N-glycosylated 124:137 arg1 transporter 96:106 Glucose transporter 4 88:108 Glucose transporter 4 (GLUT4) 88:116 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 7 31 theme fluorogenic 966:976 arg1 probes 978:983 the fluorogenic probes 962:983 the fluorogenic probes 962:983 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 3 32 theme translocation 487:499 arg1 imaging 470:476 multicolor imaging 459:476 multicolor imaging of GLUT4 translocation 459:499 Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. 27547921 7 33 theme probes 978:983 arg1 utility 951:957 the utility 947:957 the utility of the fluorogenic probes 947:983 This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization. 27547921 0 34 theme intracellular 61:73 arg1 trafficking 75:85 intracellular trafficking 61:85 intracellular trafficking 61:85 Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking. 27547921 1 35 theme N-glycosylated 124:137 arg1 protein 139:145 an N-glycosylated protein 121:145 an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation 121:220 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 1 35 theme N-glycosylated 124:137 arg1 transporter 96:106 Glucose transporter 4 88:108 Glucose transporter 4 (GLUT4) 88:116 Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. 27547921 6 36 theme GLUT4 901:905 arg1 retention 888:896 the retention 884:896 the retention of GLUT4 on the cell membrane 884:926 The results strongly suggest that the N-glycan functions in the retention of GLUT4 on the cell membrane. 27547921 6 37 theme cell 914:917 arg1 membrane 919:926 the cell membrane 910:926 the cell membrane 910:926 The results strongly suggest that the N-glycan functions in the retention of GLUT4 on the cell membrane. 26110648 6 0 theme design 940:945 arg1 strategies 947:956 design strategies 940:956 design strategies guiding HIV gp120 evolution in a rational way 940:1002 Thus, it will help design strategies guiding HIV gp120 evolution in a rational way. 26110648 1 1 theme Clade 140:144 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 2 theme mean 225:228 arg1 mfDCA 262:266 mfDCA 262:266 mfDCA 262:266 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 2 theme mean 225:228 arg1 analysis 252:259 mean field direct coupling analysis 225:259 mean field direct coupling analysis (mfDCA) 225:267 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 0 3 theme N-Glycosylation 54:68 arg1 Sites 70:74 Envelope's N-Glycosylation Sites 43:74 Envelope's N-Glycosylation Sites 43:74 The Patterns of Coevolution in Clade B HIV Envelope's N-Glycosylation Sites. 26110648 0 4 from Patterns 4:11 arg1 B 37:37 Clade B 31:37 Clade B 31:37 The Patterns of Coevolution in Clade B HIV Envelope's N-Glycosylation Sites. 26110648 2 5 theme suitable 311:318 arg1 entropy 320:326 suitable entropy 311:326 suitable entropy 311:326 This was possible for 327 positions with suitable entropy and gap content. 26110648 1 6 theme field 230:234 arg1 mfDCA 262:266 mfDCA 262:266 mfDCA 262:266 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 6 theme field 230:234 arg1 analysis 252:259 mean field direct coupling analysis 225:259 mean field direct coupling analysis (mfDCA) 225:267 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 5 7 dep map 794:796 arg1 we 798:799 we 798:799 we 798:799 The map we propose fills the gap in previous attempts to tease out sequon evolution by providing a more general molecular context. 26110648 5 8 theme general 894:900 arg1 context 912:918 a more general molecular context 887:918 a more general molecular context 887:918 The map we propose fills the gap in previous attempts to tease out sequon evolution by providing a more general molecular context. 26110648 1 9 theme direct 236:241 arg1 mfDCA 262:266 mfDCA 262:266 mfDCA 262:266 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 9 theme direct 236:241 arg1 analysis 252:259 mean field direct coupling analysis 225:259 mean field direct coupling analysis (mfDCA) 225:267 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 5 10 theme molecular 902:910 arg1 context 912:918 a more general molecular context 887:918 a more general molecular context 887:918 The map we propose fills the gap in previous attempts to tease out sequon evolution by providing a more general molecular context. 26110648 1 11 theme coupling 243:250 arg1 mfDCA 262:266 mfDCA 262:266 mfDCA 262:266 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 11 theme coupling 243:250 arg1 analysis 252:259 mean field direct coupling analysis 225:259 mean field direct coupling analysis (mfDCA) 225:267 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 3 12 dep pressure 360:367 arg1 preserve 372:379 preserve 372:379 to preserve the evolving glycan shield 369:406 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 13 theme N-glycosylation 486:500 arg1 sites 502:506 the potential N-glycosylation sites 472:506 the potential N-glycosylation sites 472:506 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 5 14 theme sequon 857:862 arg1 evolution 864:872 sequon evolution 857:872 sequon evolution 857:872 The map we propose fills the gap in previous attempts to tease out sequon evolution by providing a more general molecular context. 26110648 1 15 theme gp120 148:152 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 15 theme gp120 148:152 arg1 sites 127:131 the potential N-glycosylation sites 97:131 the potential N-glycosylation sites of HIV Clade B gp120 97:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 3 16 theme sites 502:506 arg1 rest 516:519 the rest 512:519 the rest of the structure 512:536 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 16 theme sites 502:506 arg1 majority 460:467 the majority 456:467 the majority of the potential N-glycosylation sites 456:506 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 17 theme potential 476:484 arg1 sites 502:506 the potential N-glycosylation sites 472:506 the potential N-glycosylation sites 472:506 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 18 theme evolving 385:392 arg1 glycan 394:399 the evolving glycan shield 381:406 the evolving glycan shield 381:406 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 4 19 theme core 689:692 arg1 polypeptide 694:704 the core polypeptide 685:704 the core polypeptide 685:704 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 3 20 theme structure 528:536 arg1 rest 516:519 the rest 512:519 the rest of the structure 512:536 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 20 theme structure 528:536 arg1 majority 460:467 the majority 456:467 the majority of the potential N-glycosylation sites 456:506 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 4 21 theme evolving 635:642 arg1 related 674:680 related 674:680 related 674:680 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 4 21 theme evolving 635:642 arg1 glycan 644:649 the evolving glycan shield 631:656 the evolving glycan shield 631:656 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 0 22 theme Coevolution 16:26 arg1 Patterns 4:11 The Patterns 0:11 The Patterns of Coevolution in Clade B 0:37 The Patterns of Coevolution in Clade B HIV Envelope's N-Glycosylation Sites. 26110648 1 23 theme B 146:146 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 24 theme coevolution 174:184 arg1 network 186:192 the coevolution network 170:192 the coevolution network of the protein structure 170:217 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 0 25 theme Clade 31:35 arg1 B 37:37 Clade B 31:37 Clade B 31:37 The Patterns of Coevolution in Clade B HIV Envelope's N-Glycosylation Sites. 26110648 6 26 theme gp120 970:974 arg1 evolution 976:984 HIV gp120 evolution 966:984 HIV gp120 evolution 966:984 Thus, it will help design strategies guiding HIV gp120 evolution in a rational way. 26110648 6 27 theme rational 991:998 arg1 way 1000:1002 a rational way 989:1002 a rational way 989:1002 Thus, it will help design strategies guiding HIV gp120 evolution in a rational way. 26110648 1 28 gly N-glycosylation 111:125 arg2 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 28 gly N-glycosylation 111:125 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 28 gly N-glycosylation 111:125 arg2 sites 127:131 the potential N-glycosylation sites 97:131 the potential N-glycosylation sites of HIV Clade B gp120 97:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 3 29 dep glycan 394:399 arg1 shield 401:406 shield 401:406 shield 401:406 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 4 30 theme antibody 606:613 arg1 neutralization 615:628 antibody neutralization 606:628 antibody neutralization 606:628 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 3 31 theme pressure 360:367 arg1 Indications 345:355 Indications 345:355 Indications of pressure to preserve the evolving glycan shield 345:406 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 3 32 theme strong 428:433 arg1 dependencies 435:446 as well as strong dependencies 417:446 as well as strong dependencies 417:446 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 1 33 theme potential 101:109 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 33 theme potential 101:109 arg1 sites 127:131 the potential N-glycosylation sites 97:131 the potential N-glycosylation sites of HIV Clade B gp120 97:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 4 34 dep mainly 577:582 arg1 adaptation 587:596 an adaptation 584:596 an adaptation against antibody neutralization 584:628 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 4 35 dep glycan 644:649 arg1 shield 651:656 shield 651:656 shield 651:656 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 4 36 from changes 758:764 arg1 N-glycosylation 773:787 the N-glycosylation 769:787 the N-glycosylation 769:787 These findings indicate that although mainly an adaptation against antibody neutralization, the evolving glycan shield is structurally related to the core polypeptide, which, thus, is also under pressure to reflect the changes in the N-glycosylation. 26110648 1 37 theme N-glycosylation 111:125 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 37 theme N-glycosylation 111:125 arg1 sites 127:131 the potential N-glycosylation sites 97:131 the potential N-glycosylation sites of HIV Clade B gp120 97:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 3 38 gly N-glycosylation 486:500 arg2 sites 502:506 the potential N-glycosylation sites 472:506 the potential N-glycosylation sites 472:506 Indications of pressure to preserve the evolving glycan shield are seen as well as strong dependencies between the majority of the potential N-glycosylation sites and the rest of the structure. 26110648 1 39 theme sites 127:131 arg1 co-evolution 81:92 The co-evolution 77:92 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 77:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 1 40 theme protein 201:207 arg1 structure 209:217 the protein structure 197:217 the protein structure 197:217 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 6 41 theme HIV 966:968 arg1 evolution 976:984 HIV gp120 evolution 966:984 HIV gp120 evolution 966:984 Thus, it will help design strategies guiding HIV gp120 evolution in a rational way. 26110648 2 42 theme gap 332:334 arg1 content 336:342 gap content 332:342 gap content 332:342 This was possible for 327 positions with suitable entropy and gap content. 26110648 1 43 theme structure 209:217 arg1 network 186:192 the coevolution network 170:192 the coevolution network of the protein structure 170:217 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 26110648 2 44 with possible 279:286 arg1 entropy 320:326 suitable entropy 311:326 suitable entropy 311:326 This was possible for 327 positions with suitable entropy and gap content. 26110648 2 44 with possible 279:286 arg1 content 336:342 gap content 332:342 gap content 332:342 This was possible for 327 positions with suitable entropy and gap content. 26110648 1 45 theme HIV 136:138 arg1 gp120 148:152 HIV Clade B gp120 136:152 HIV Clade B gp120 136:152 The co-evolution of the potential N-glycosylation sites of HIV Clade B gp120 was mapped onto the coevolution network of the protein structure using mean field direct coupling analysis (mfDCA). 28039392 0 0 theme family 87:92 arg1 α1,3/4-fucosidase 37:53 tomato α1,3/4-fucosidase 30:53 tomato α1,3/4-fucosidase 30:53 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 0 0 theme family 87:92 arg1 member 58:63 a member 56:63 a member of glycosyl hydrolase family 29 56:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 4 1 theme pyridylaminated 767:781 arg1 Fucα1-3GlcNAc 783:795 pyridylaminated Fucα1-3GlcNAc 767:795 pyridylaminated Fucα1-3GlcNAc 767:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 6 2 theme Molecular 988:996 arg1 modelling 1001:1009 Molecular 3D modelling 988:1009 Molecular 3D modelling of α-Fuc 988:1018 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 1 3 theme acidic 220:225 arg1 α-fucosidase 227:238 an acidic α-fucosidase 217:238 an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1) 217:287 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 1 3 theme acidic 220:225 arg1 gene 189:192 a gene 187:192 a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans 187:358 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 0 4 theme hydrolase 77:85 arg1 family 87:92 glycosyl hydrolase family 29 68:95 glycosyl hydrolase family 29 68:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 6 5 theme 3D 998:999 arg1 modelling 1001:1009 Molecular 3D modelling 988:1009 Molecular 3D modelling of α-Fuc 988:1018 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 5 6 theme N-glycans 977:985 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 7 from residue 601:607 arg1 Fucα1-3GlcNAc 783:795 pyridylaminated Fucα1-3GlcNAc 767:795 pyridylaminated Fucα1-3GlcNAc 767:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 7 from residue 601:607 arg1 LNFP 612:615 LNFP III 612:619 LNFP III 612:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 7 from residue 601:607 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 7 from residue 601:607 arg1 I 735:735 LNFP I 730:735 LNFP I 730:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 8 from N-glycans 684:692 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 8 from N-glycans 684:692 arg1 residue 756:762 the α1,3-fucose residue 740:762 the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc 740:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 8 from N-glycans 684:692 arg1 residue 601:607 the non-reducing terminal α1,3-fucose residue 563:607 the non-reducing terminal α1,3-fucose residue on LNFP III 563:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 8 from N-glycans 684:692 arg1 residue 719:725 the α1,2-fucose residue 703:725 the α1,2-fucose residue on LNFP I 703:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 8 from N-glycans 684:692 arg1 residues 637:644 α1,4-fucose residues 625:644 α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans 625:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 9 theme complex-type 671:682 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 3 10 contain has 505:507 arg2 pH 520:521 an optimum pH 509:521 an optimum pH around 4.5 509:532 rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. 28039392 3 10 contain has 505:507 arg1 Sl-1 478:481 rFuc'ase Sl-1 469:481 rFuc'ase Sl-1 469:481 rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. 28039392 2 11 theme expression 450:459 arg1 system 461:466 a baculovirus-insect cell expression system 424:466 a baculovirus-insect cell expression system 424:466 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 4 12 from residue 756:762 arg1 Fucα1-3GlcNAc 783:795 pyridylaminated Fucα1-3GlcNAc 767:795 pyridylaminated Fucα1-3GlcNAc 767:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 12 from residue 756:762 arg1 LNFP 612:615 LNFP III 612:619 LNFP III 612:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 12 from residue 756:762 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 12 from residue 756:762 arg1 I 735:735 LNFP I 730:735 LNFP I 730:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 13 theme plant 665:669 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 3 14 theme ase 474:476 arg1 Sl-1 478:481 rFuc'ase Sl-1 469:481 rFuc'ase Sl-1 469:481 rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. 28039392 5 15 theme α-Fuc 837:841 arg1 S1-1 847:850 this tomato α-Fuc'ase S1-1 825:850 this tomato α-Fuc'ase S1-1 825:850 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 2 16 theme cell 445:448 arg1 system 461:466 a baculovirus-insect cell expression system 424:466 a baculovirus-insect cell expression system 424:466 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 7 17 dep residues 1189:1196 arg1 residues 1189:1196 residues Asp193 and Glu237 1189:1214 residues Asp193 and Glu237 1189:1214 infantis (Blon_2336) indicated that residues Asp193 and Glu237 might be important for substrate binding. 28039392 7 17 dep residues 1189:1196 arg1 Glu237 1209:1214 Glu237 1209:1214 Glu237 1209:1214 infantis (Blon_2336) indicated that residues Asp193 and Glu237 might be important for substrate binding. 28039392 7 17 dep residues 1189:1196 arg1 Asp193 1198:1203 Asp193 1198:1203 Asp193 1198:1203 infantis (Blon_2336) indicated that residues Asp193 and Glu237 might be important for substrate binding. 28039392 7 18 theme substrate 1239:1247 arg1 binding 1249:1255 substrate binding 1239:1255 substrate binding 1239:1255 infantis (Blon_2336) indicated that residues Asp193 and Glu237 might be important for substrate binding. 28039392 2 19 theme baculovirus-insect 426:443 arg1 system 461:466 a baculovirus-insect cell expression system 424:466 a baculovirus-insect cell expression system 424:466 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 5 20 theme core 876:879 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 21 from epitopes 653:660 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 22 theme Lea 649:651 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 23 theme terminal 580:587 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 23 theme terminal 580:587 arg1 residue 601:607 the non-reducing terminal α1,3-fucose residue 563:607 the non-reducing terminal α1,3-fucose residue on LNFP III 563:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 0 24 theme Molecular 0:8 arg1 characterization 10:25 Molecular characterization 0:25 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 0:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 6 25 theme homologous 1094:1103 arg1 α-fucosidase 1105:1116 a homologous α-fucosidase 1092:1116 a homologous α-fucosidase from Bifidobacterium longum subsp 1092:1150 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 4 26 theme α1,3-fucose 744:754 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 26 theme α1,3-fucose 744:754 arg1 residue 756:762 the α1,3-fucose residue 740:762 the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc 740:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 5 27 theme ase 843:845 arg1 S1-1 847:850 this tomato α-Fuc'ase S1-1 825:850 this tomato α-Fuc'ase S1-1 825:850 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 28 theme non-reducing 567:578 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 28 theme non-reducing 567:578 arg1 residue 601:607 the non-reducing terminal α1,3-fucose residue 563:607 the non-reducing terminal α1,3-fucose residue on LNFP III 563:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 0 29 theme tomato 30:35 arg1 α1,3/4-fucosidase 37:53 tomato α1,3/4-fucosidase 30:53 tomato α1,3/4-fucosidase 30:53 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 0 29 theme tomato 30:35 arg1 member 58:63 a member 56:63 a member of glycosyl hydrolase family 29 56:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 5 30 theme [Manβ1-4 908:915 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 2 31 theme ase 379:381 arg1 S1-1 398:401 rFuc'ase S1-1 389:401 rFuc'ase S1-1 389:401 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 2 31 theme ase 379:381 arg1 S1-1 383:386 Recombinant α-Fuc'ase S1-1 361:386 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) 361:402 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 4 32 theme epitopes 653:660 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 32 theme epitopes 653:660 arg1 residue 756:762 the α1,3-fucose residue 740:762 the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc 740:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 32 theme epitopes 653:660 arg1 residue 601:607 the non-reducing terminal α1,3-fucose residue 563:607 the non-reducing terminal α1,3-fucose residue on LNFP III 563:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 32 theme epitopes 653:660 arg1 residue 719:725 the α1,2-fucose residue 703:725 the α1,2-fucose residue on LNFP I 703:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 32 theme epitopes 653:660 arg1 residues 637:644 α1,4-fucose residues 625:644 α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans 625:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 2 33 theme α-Fuc 373:377 arg1 S1-1 398:401 rFuc'ase S1-1 389:401 rFuc'ase S1-1 389:401 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 2 33 theme α-Fuc 373:377 arg1 S1-1 383:386 Recombinant α-Fuc'ase S1-1 361:386 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) 361:402 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 6 34 with comparison 1076:1085 arg1 α-fucosidase 1105:1116 a homologous α-fucosidase 1092:1116 a homologous α-fucosidase from Bifidobacterium longum subsp 1092:1150 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 0 35 theme complex 134:140 arg1 N-glycans 147:155 plant complex type N-glycans 128:155 plant complex type N-glycans 128:155 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 1 36 theme plant 331:335 arg1 N-glycans 350:358 plant complex-type N-glycans 331:358 plant complex-type N-glycans 331:358 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 5 37 theme Xylβ1-2 917:923 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 6 38 from subsp 1146:1150 arg1 α-fucosidase 1105:1116 a homologous α-fucosidase 1092:1116 a homologous α-fucosidase from Bifidobacterium longum subsp 1092:1150 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 2 39 theme Recombinant 361:371 arg1 S1-1 398:401 rFuc'ase S1-1 389:401 rFuc'ase S1-1 389:401 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 2 39 theme Recombinant 361:371 arg1 S1-1 383:386 Recombinant α-Fuc'ase S1-1 361:386 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) 361:402 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 0 40 theme α1,3/4-fucosidase 37:53 arg1 characterization 10:25 Molecular characterization 0:25 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 0:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 0 41 theme plant 128:132 arg1 N-glycans 147:155 plant complex type N-glycans 128:155 plant complex type N-glycans 128:155 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 1 42 theme complex-type 337:348 arg1 N-glycans 350:358 plant complex-type N-glycans 331:358 plant complex-type N-glycans 331:358 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 1 43 from gene 189:192 arg1 tomato 197:202 tomato 197:202 tomato 197:202 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 2 44 theme ase 394:396 arg1 S1-1 398:401 rFuc'ase S1-1 389:401 rFuc'ase S1-1 389:401 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 2 44 theme ase 394:396 arg1 S1-1 383:386 Recombinant α-Fuc'ase S1-1 361:386 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) 361:402 Recombinant α-Fuc'ase S1-1 (rFuc'ase S1-1) was expressed using a baculovirus-insect cell expression system. 28039392 1 45 theme ase 279:281 arg1 S1-1 283:286 α-Fuc'ase S1-1 273:286 α-Fuc'ase S1-1 273:286 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 1 45 theme ase 279:281 arg1 LOC101254568 241:252 LOC101254568 241:252 LOC101254568 241:252 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 0 46 theme N-glycans 147:155 arg1 degradation 113:123 the degradation 109:123 the degradation of plant complex type N-glycans 109:155 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 1 47 theme N-glycans 350:358 arg1 turnover 319:326 the turnover 315:326 the turnover of plant complex-type N-glycans 315:358 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 5 48 theme GlcNAcβ1-4 925:934 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 6 49 theme ase 1020:1022 arg1 Sl-1 1024:1027 ase Sl-1 1020:1027 ase Sl-1 1020:1027 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 0 50 theme type 142:145 arg1 N-glycans 147:155 plant complex type N-glycans 128:155 plant complex type N-glycans 128:155 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 5 51 theme Fucα1-3 936:942 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 52 theme LNFP 730:733 arg1 I 735:735 LNFP I 730:735 LNFP I 730:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 5 53 theme complex-type 964:975 arg1 N-glycans 977:985 plant complex-type N-glycans 958:985 plant complex-type N-glycans 958:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 54 from residues 637:644 arg1 Fucα1-3GlcNAc 783:795 pyridylaminated Fucα1-3GlcNAc 767:795 pyridylaminated Fucα1-3GlcNAc 767:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 54 from residues 637:644 arg1 LNFP 612:615 LNFP III 612:619 LNFP III 612:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 54 from residues 637:644 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 54 from residues 637:644 arg1 I 735:735 LNFP I 730:735 LNFP I 730:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 5 55 theme penta-oligosaccharide 881:901 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 56 from residue 719:725 arg1 Fucα1-3GlcNAc 783:795 pyridylaminated Fucα1-3GlcNAc 767:795 pyridylaminated Fucα1-3GlcNAc 767:795 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 56 from residue 719:725 arg1 LNFP 612:615 LNFP III 612:619 LNFP III 612:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 56 from residue 719:725 arg1 N-glycans 684:692 plant complex-type N-glycans 665:692 plant complex-type N-glycans 665:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 56 from residue 719:725 arg1 I 735:735 LNFP I 730:735 LNFP I 730:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 5 57 theme tomato 830:835 arg1 S1-1 847:850 this tomato α-Fuc'ase S1-1 825:850 this tomato α-Fuc'ase S1-1 825:850 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 6 58 theme longum 1139:1144 arg1 subsp 1146:1150 Bifidobacterium longum subsp 1123:1150 Bifidobacterium longum subsp 1123:1150 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 5 59 theme unit 903:906 arg1 GlcNAc-PA 944:952 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] 872:953 the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans 872:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 5 60 theme plant 958:962 arg1 N-glycans 977:985 plant complex-type N-glycans 958:985 plant complex-type N-glycans 958:985 Furthermore, we found that this tomato α-Fuc'ase S1-1 was inactive toward the core penta-oligosaccharide unit [Manβ1-4(Xylβ1-2)GlcNAcβ1-4(Fucα1-3)GlcNAc-PA] of plant complex-type N-glycans. 28039392 4 61 theme α1,4-fucose 625:635 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 61 theme α1,4-fucose 625:635 arg1 residues 637:644 α1,4-fucose residues 625:644 α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans 625:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 6 62 theme α-Fuc 1014:1018 arg1 modelling 1001:1009 Molecular 3D modelling 988:1009 Molecular 3D modelling of α-Fuc 988:1018 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 0 63 theme glycosyl 68:75 arg1 family 87:92 glycosyl hydrolase family 29 68:95 glycosyl hydrolase family 29 68:95 Molecular characterization of tomato α1,3/4-fucosidase, a member of glycosyl hydrolase family 29 involved in the degradation of plant complex type N-glycans. 28039392 1 64 dep α-fucosidase 227:238 arg1 S1-1 283:286 α-Fuc'ase S1-1 273:286 α-Fuc'ase S1-1 273:286 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 1 64 dep α-fucosidase 227:238 arg1 Solyc03g006980 257:270 Solyc03g006980 257:270 Solyc03g006980 257:270 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 1 64 dep α-fucosidase 227:238 arg1 LOC101254568 241:252 LOC101254568 241:252 LOC101254568 241:252 In this study, we identified a gene in tomato that encodes an acidic α-fucosidase (LOC101254568 or Solyc03g006980, α-Fuc'ase S1-1), which may be involved in the turnover of plant complex-type N-glycans. 28039392 4 65 theme α1,2-fucose 707:717 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 65 theme α1,2-fucose 707:717 arg1 residue 719:725 the α1,2-fucose residue 703:725 the α1,2-fucose residue on LNFP I 703:735 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 3 66 from kDa 489:491 arg1 size 496:499 size 496:499 size 496:499 rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. 28039392 6 67 theme structure/sequence 1033:1050 arg1 interpretation 1052:1065 structure/sequence interpretation 1033:1065 structure/sequence interpretation 1033:1065 Molecular 3D modelling of α-Fuc'ase Sl-1 and structure/sequence interpretation based on comparison with a homologous α-fucosidase from Bifidobacterium longum subsp. 28039392 3 68 theme optimum 512:518 arg1 pH 520:521 an optimum pH 509:521 an optimum pH around 4.5 509:532 rFuc'ase Sl-1 is 55 kDa in size and has an optimum pH around 4.5. 28039392 4 69 theme α1,3-fucose 589:599 arg1 epitopes 653:660 Lea epitopes 649:660 Lea epitopes on plant complex-type N-glycans 649:692 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 28039392 4 69 theme α1,3-fucose 589:599 arg1 residue 601:607 the non-reducing terminal α1,3-fucose residue 563:607 the non-reducing terminal α1,3-fucose residue on LNFP III 563:619 It substantially hydrolyzed the non-reducing terminal α1,3-fucose residue on LNFP III and α1,4-fucose residues of Lea epitopes on plant complex-type N-glycans, but not the α1,2-fucose residue on LNFP I or the α1,3-fucose residue on pyridylaminated Fucα1-3GlcNAc. 27000970 0 0 theme α-amylase 108:116 arg1 inactivation 51:62 mechanism-based inactivation 35:62 mechanism-based inactivation 35:62 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 0 0 theme α-amylase 108:116 arg1 analysis 79:86 structural analysis 68:86 structural analysis 68:86 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 1 1 theme covalent 170:177 arg1 inhibitors 179:188 selective, mechanism-based covalent inhibitors 143:188 inhibitors 179:188 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 2 theme analog 281:286 arg1 epi-cyclophellitol 299:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 0 3 theme pancreatic 97:106 arg1 α-amylase 108:116 human pancreatic α-amylase 91:116 human pancreatic α-amylase 91:116 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 1 4 theme α-glucosyl 288:297 arg1 epi-cyclophellitol 299:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 3 5 from OH4 699:701 arg1 sugar 727:731 the nonreducing end sugar 707:731 the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo 707:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 1 6 theme time 408:411 arg1 dependence 413:422 the time dependence 404:422 the time dependence of its inhibition 404:440 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 7 theme epi-cyclophellitol 299:316 arg1 synthesis 251:259 the chemoenzymatic synthesis 232:259 the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol 232:316 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 2 8 theme bond 582:585 arg1 formation 558:566 formation 558:566 formation of a covalent bond to the catalytic nucleophile D197 558:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 3 9 theme fluorescent 766:776 arg1 tags 778:781 fluorescent tags 766:781 fluorescent tags in generating probes for localization and quantitation of amylase in vivo 766:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 2 10 theme covalent 573:580 arg1 bond 582:585 a covalent bond 571:585 a covalent bond to the catalytic nucleophile D197 571:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 3 11 theme tags 778:781 arg1 attachment 752:761 attachment 752:761 attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo 752:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 2 12 theme X-ray 443:447 arg1 analysis 466:473 X-ray crystallographic analysis 443:473 X-ray crystallographic analysis of the covalent derivative so formed 443:510 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 1 13 theme human 193:197 arg1 α-amylase 210:218 human pancreatic α-amylase 193:218 human pancreatic α-amylase 193:218 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 14 theme inhibition 431:440 arg1 dependence 413:422 the time dependence 404:422 the time dependence of its inhibition 404:440 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 15 with reaction 350:357 arg1 α-amylase 381:389 human pancreatic α-amylase 364:389 human pancreatic α-amylase 364:389 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 16 theme pancreatic 199:208 arg1 α-amylase 210:218 human pancreatic α-amylase 193:218 human pancreatic α-amylase 193:218 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 0 17 theme Glucosyl 0:7 arg1 epi-cyclophellitol 9:26 Glucosyl epi-cyclophellitol 0:26 Glucosyl epi-cyclophellitol 0:26 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 1 18 theme α-amylase 210:218 arg1 inhibitors 179:188 selective, mechanism-based covalent inhibitors 143:188 inhibitors 179:188 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 0 19 theme mechanism-based 35:49 arg1 inactivation 51:62 mechanism-based inactivation 35:62 mechanism-based inactivation 35:62 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 3 20 theme active 674:679 arg1 site 681:684 the active site 670:684 the active site 670:684 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 2 21 theme derivative 491:500 arg1 analysis 466:473 X-ray crystallographic analysis 443:473 X-ray crystallographic analysis of the covalent derivative so formed 443:510 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 1 22 theme stoichiometric 335:348 arg1 reaction 350:357 its stoichiometric reaction 331:357 its stoichiometric reaction with human pancreatic α-amylase 331:389 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 2 23 theme nucleophile 604:614 arg1 D197 616:619 the catalytic nucleophile D197 590:619 the catalytic nucleophile D197 590:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 2 24 theme covalent 482:489 arg1 derivative 491:500 the covalent derivative 478:500 the covalent derivative so formed 478:510 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 2 25 theme active 541:546 arg1 site 548:551 the active site 537:551 the active site 537:551 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 2 26 theme catalytic 594:602 arg1 nucleophile 604:614 the catalytic nucleophile 590:614 the catalytic nucleophile D197 590:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 2 27 from site 548:551 arg1 reaction 525:532 its reaction 521:532 its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197 521:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 1 28 theme chemoenzymatic 236:249 arg1 synthesis 251:259 the chemoenzymatic synthesis 232:259 the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol 232:316 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 29 theme human 364:368 arg1 α-amylase 381:389 human pancreatic α-amylase 364:389 human pancreatic α-amylase 364:389 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 0 30 theme structural 68:77 arg1 analysis 79:86 structural analysis 68:86 structural analysis 68:86 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 1 31 theme pancreatic 370:379 arg1 α-amylase 381:389 human pancreatic α-amylase 364:389 human pancreatic α-amylase 364:389 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 3 32 with interactions 652:663 arg1 site 681:684 the active site 670:684 the active site 670:684 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 1 33 theme search 132:137 arg1 part 122:125 part 122:125 part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase 122:218 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 3 34 theme nonreducing 711:721 arg1 sugar 727:731 the nonreducing end sugar 707:731 the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo 707:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 1 35 theme selective 143:151 arg1 inhibitors 179:188 selective, mechanism-based covalent inhibitors 143:188 inhibitors 179:188 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 0 36 theme human 91:95 arg1 α-amylase 108:116 human pancreatic α-amylase 91:116 human pancreatic α-amylase 91:116 Glucosyl epi-cyclophellitol allows mechanism-based inactivation and structural analysis of human pancreatic α-amylase. 27000970 3 37 theme end 723:725 arg1 sugar 727:731 the nonreducing end sugar 707:731 the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo 707:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 3 38 theme good 738:741 arg1 site 743:746 a good site 736:746 a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo 736:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 2 39 with reaction 525:532 arg1 formation 558:566 formation 558:566 formation of a covalent bond to the catalytic nucleophile D197 558:619 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 2 40 theme crystallographic 449:464 arg1 analysis 466:473 X-ray crystallographic analysis 443:473 X-ray crystallographic analysis of the covalent derivative so formed 443:510 X-ray crystallographic analysis of the covalent derivative so formed confirms its reaction at the active site with formation of a covalent bond to the catalytic nucleophile D197. 27000970 3 41 theme amylase 841:847 arg1 in vivo 849:855 amylase in vivo 841:855 amylase in vivo 841:855 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 3 42 theme in vivo 849:855 arg1 quantitation 825:836 quantitation 825:836 quantitation 825:836 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 3 42 theme in vivo 849:855 arg1 localization 808:819 localization 808:819 localization 808:819 The structure illuminates the interactions with the active site and confirms OH4' on the nonreducing end sugar as a good site for attachment of fluorescent tags in generating probes for localization and quantitation of amylase in vivo. 27000970 1 43 theme mechanism-based 154:168 arg1 inhibitors 179:188 selective, mechanism-based covalent inhibitors 143:188 inhibitors 179:188 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 27000970 1 44 theme disaccharide 268:279 arg1 epi-cyclophellitol 299:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 the disaccharide analog α-glucosyl epi-cyclophellitol 264:316 As part of a search for selective, mechanism-based covalent inhibitors of human pancreatic α-amylase we describe the chemoenzymatic synthesis of the disaccharide analog α-glucosyl epi-cyclophellitol, demonstrate its stoichiometric reaction with human pancreatic α-amylase and evaluate the time dependence of its inhibition. 26774377 3 0 theme average 323:329 arg1 weight 341:346 an average molecular weight 320:346 an average molecular weight of 3.22 × 10(4)Da 320:364 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 4 1 theme periodate 480:488 arg1 degradation 506:516 periodate oxidation-smith degradation 480:516 periodate oxidation-smith degradation 480:516 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 7 2 from acidity 1090:1096 arg1 serum 1139:1143 serum 1139:1143 serum 1139:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 6 3 theme gastric 857:863 arg1 ulcer 865:869 the ethanol-induced gastric ulcer 837:869 the ethanol-induced gastric ulcer 837:869 To study the protective effects of LEP-2a on the ethanol-induced gastric ulcer in mice, LEP-2a (100, 200 and 400mg/kg/d) was given to mice by gavage for 2 weeks. 26774377 0 4 theme expolysaccharide 77:92 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 0 4 theme expolysaccharide 77:92 arg1 effects 25:31 preventive effects 14:31 preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp 14:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 7 5 from volume 1062:1067 arg1 serum 1139:1143 serum 1139:1143 serum 1139:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 4 6 theme chemical 412:419 arg1 analysis 470:477 methylation analysis 458:477 methylation analysis 458:477 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 6 theme chemical 412:419 arg1 degradation 506:516 periodate oxidation-smith degradation 480:516 periodate oxidation-smith degradation 480:516 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 6 theme chemical 412:419 arg1 methods 439:445 chemical and spectroscopic methods 412:445 chemical and spectroscopic methods 412:445 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 6 theme chemical 412:419 arg1 spectroscopy 528:539 infrared spectroscopy 519:539 infrared spectroscopy 519:539 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 6 theme chemical 412:419 arg1 analysis 549:556 NMR analysis 545:556 NMR analysis 545:556 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 7 7 theme pepsin 1031:1036 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 7 theme pepsin 1031:1036 arg1 activity 1038:1045 pepsin activity 1031:1045 pepsin activity 1031:1045 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 6 8 theme LEP-2a 827:832 arg1 effects 816:822 the protective effects 801:822 the protective effects of LEP-2a on the ethanol-induced gastric ulcer 801:869 To study the protective effects of LEP-2a on the ethanol-induced gastric ulcer in mice, LEP-2a (100, 200 and 400mg/kg/d) was given to mice by gavage for 2 weeks. 26774377 7 9 theme malondialdehyde 1106:1120 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 9 theme malondialdehyde 1106:1120 arg1 content 1128:1134 the malondialdehyde (MDA) content 1102:1134 the malondialdehyde (MDA) content in serum 1102:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 0 10 from Structure 0:8 arg1 sp 107:108 Lachnum sp 99:108 Lachnum sp 99:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 0 11 theme Lachnum 99:105 arg1 sp 107:108 Lachnum sp 99:108 Lachnum sp 99:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 4 12 theme methylation 458:468 arg1 analysis 470:477 methylation analysis 458:477 methylation analysis 458:477 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 5 13 theme branch 624:629 arg1 chain 631:635 chain 631:635 chain 631:635 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 4 14 theme LEP-2a 384:389 arg1 structure 371:379 The structure 367:379 The structure of LEP-2a 367:389 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 7 15 from content 1128:1134 arg1 serum 1139:1143 serum 1139:1143 serum 1139:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 9 16 theme significant 1260:1270 arg1 effect 1283:1288 a significant preventive effect 1258:1288 a significant preventive effect 1258:1288 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 2 17 dep purified 168:175 arg1 LEP 159:161 LEP 159:161 LEP 159:161 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 6 18 theme ethanol-induced 841:855 arg1 ulcer 865:869 the ethanol-induced gastric ulcer 837:869 the ethanol-induced gastric ulcer 837:869 To study the protective effects of LEP-2a on the ethanol-induced gastric ulcer in mice, LEP-2a (100, 200 and 400mg/kg/d) was given to mice by gavage for 2 weeks. 26774377 7 19 theme bleeding 1015:1022 arg1 volume 1062:1067 gastric juice volume 1048:1067 gastric juice volume 1048:1067 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 19 theme bleeding 1015:1022 arg1 content 1128:1134 the malondialdehyde (MDA) content 1102:1134 the malondialdehyde (MDA) content in serum 1102:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 19 theme bleeding 1015:1022 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 19 theme bleeding 1015:1022 arg1 activity 1038:1045 pepsin activity 1031:1045 pepsin activity 1031:1045 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 19 theme bleeding 1015:1022 arg1 acidity 1090:1096 gastric juice total acidity 1070:1096 gastric juice total acidity 1070:1096 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 5 20 theme backbone 748:755 arg1 chain 757:761 the backbone chain 744:761 the backbone chain 744:761 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 9 21 theme above 1217:1221 arg1 findings 1223:1230 The above findings 1213:1230 The above findings 1213:1230 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 3 22 with component 305:313 arg1 weight 341:346 an average molecular weight 320:346 an average molecular weight of 3.22 × 10(4)Da 320:364 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 3 23 theme Da 363:364 arg1 weight 341:346 an average molecular weight 320:346 an average molecular weight of 3.22 × 10(4)Da 320:364 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 3 24 theme molecular 331:339 arg1 weight 341:346 an average molecular weight 320:346 an average molecular weight of 3.22 × 10(4)Da 320:364 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 5 25 theme 1→6 603:605 arg1 1→3 596:598 1→3 596:598 1→3 596:598 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 5 25 theme 1→6 603:605 arg1 -β-D-Glcp 607:615 (1→6)-β-D-Glcp 602:615 (1→6)-β-D-Glcp 602:615 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 7 26 theme total 1084:1088 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 26 theme total 1084:1088 arg1 acidity 1090:1096 gastric juice total acidity 1070:1096 gastric juice total acidity 1070:1096 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 27 theme ulcer 1009:1013 arg1 volume 1062:1067 gastric juice volume 1048:1067 gastric juice volume 1048:1067 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 27 theme ulcer 1009:1013 arg1 content 1128:1134 the malondialdehyde (MDA) content 1102:1134 the malondialdehyde (MDA) content in serum 1102:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 27 theme ulcer 1009:1013 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 27 theme ulcer 1009:1013 arg1 activity 1038:1045 pepsin activity 1031:1045 pepsin activity 1031:1045 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 27 theme ulcer 1009:1013 arg1 acidity 1090:1096 gastric juice total acidity 1070:1096 gastric juice total acidity 1070:1096 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 9 28 theme preventive 1272:1281 arg1 effect 1283:1288 a significant preventive effect 1258:1288 a significant preventive effect 1258:1288 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 6 29 from effects 816:822 arg1 ulcer 865:869 the ethanol-induced gastric ulcer 837:869 the ethanol-induced gastric ulcer 837:869 To study the protective effects of LEP-2a on the ethanol-induced gastric ulcer in mice, LEP-2a (100, 200 and 400mg/kg/d) was given to mice by gavage for 2 weeks. 26774377 9 30 theme gastric 1318:1324 arg1 ulcer 1326:1330 the ethanol-induced gastric ulcer 1298:1330 the ethanol-induced gastric ulcer 1298:1330 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 5 31 theme β-1,6-glycosidic 766:781 arg1 linkage 783:789 β-1,6-glycosidic linkage 766:789 β-1,6-glycosidic linkage 766:789 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 7 32 from activity 1038:1045 arg1 serum 1139:1143 serum 1139:1143 serum 1139:1143 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 8 33 theme superoxide 1161:1170 arg1 Meanwhile 1146:1154 Meanwhile 1146:1154 Meanwhile 1146:1154 Meanwhile, the superoxide dismutase (SOD) increased significantly. 26774377 8 33 theme superoxide 1161:1170 arg1 SOD 1183:1185 SOD 1183:1185 SOD 1183:1185 Meanwhile, the superoxide dismutase (SOD) increased significantly. 26774377 8 33 theme superoxide 1161:1170 arg1 dismutase 1172:1180 the superoxide dismutase 1157:1180 the superoxide dismutase (SOD) 1157:1186 Meanwhile, the superoxide dismutase (SOD) increased significantly. 26774377 0 34 theme preventive 14:23 arg1 effects 25:31 preventive effects 14:31 preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp 14:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 5 35 theme d-glucopyranosyl 656:671 arg1 residues 673:680 two d-glucopyranosyl residues 652:680 two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage 652:789 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 4 36 theme spectroscopic 425:437 arg1 analysis 470:477 methylation analysis 458:477 methylation analysis 458:477 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 36 theme spectroscopic 425:437 arg1 degradation 506:516 periodate oxidation-smith degradation 480:516 periodate oxidation-smith degradation 480:516 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 36 theme spectroscopic 425:437 arg1 methods 439:445 chemical and spectroscopic methods 412:445 chemical and spectroscopic methods 412:445 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 36 theme spectroscopic 425:437 arg1 spectroscopy 528:539 infrared spectroscopy 519:539 infrared spectroscopy 519:539 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 36 theme spectroscopic 425:437 arg1 analysis 549:556 NMR analysis 545:556 NMR analysis 545:556 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 0 37 theme ethanol-induced 41:55 arg1 ulcer 65:69 ethanol-induced gastric ulcer 41:69 ethanol-induced gastric ulcer 41:69 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 2 38 theme column 240:245 arg1 chromatography 247:260 Sepharose CL-6B column chromatography 224:260 Sepharose CL-6B column chromatography 224:260 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 2 39 theme CL-6B 234:238 arg1 chromatography 247:260 Sepharose CL-6B column chromatography 224:260 Sepharose CL-6B column chromatography 224:260 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 9 40 contain had 1254:1256 arg1 LEP-2a 1247:1252 LEP-2a 1247:1252 LEP-2a 1247:1252 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 9 40 contain had 1254:1256 arg2 effect 1283:1288 a significant preventive effect 1258:1288 a significant preventive effect 1258:1288 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 7 41 theme gastric 1048:1054 arg1 volume 1062:1067 gastric juice volume 1048:1067 gastric juice volume 1048:1067 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 41 theme gastric 1048:1054 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 4 42 theme infrared 519:526 arg1 spectroscopy 528:539 infrared spectroscopy 519:539 infrared spectroscopy 519:539 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 6 43 theme protective 805:814 arg1 effects 816:822 the protective effects 801:822 the protective effects of LEP-2a on the ethanol-induced gastric ulcer 801:869 To study the protective effects of LEP-2a on the ethanol-induced gastric ulcer in mice, LEP-2a (100, 200 and 400mg/kg/d) was given to mice by gavage for 2 weeks. 26774377 2 44 theme Sepharose 224:232 arg1 chromatography 247:260 Sepharose CL-6B column chromatography 224:260 Sepharose CL-6B column chromatography 224:260 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 3 45 dep 10 358:359 arg1 4 361:361 4 361:361 4 361:361 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 1 46 theme extracellular 114:126 arg1 polysaccharide 128:141 An extracellular polysaccharide 111:141 An extracellular polysaccharide of Lachnum sp 111:155 An extracellular polysaccharide of Lachnum sp. 26774377 0 47 theme gastric 57:63 arg1 ulcer 65:69 ethanol-induced gastric ulcer 41:69 ethanol-induced gastric ulcer 41:69 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 4 48 theme NMR 545:547 arg1 analysis 549:556 NMR analysis 545:556 NMR analysis 545:556 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 4 49 theme oxidation-smith 490:504 arg1 degradation 506:516 periodate oxidation-smith degradation 480:516 periodate oxidation-smith degradation 480:516 The structure of LEP-2a was characterized by chemical and spectroscopic methods, including methylation analysis, periodate oxidation-smith degradation, infrared spectroscopy and NMR analysis. 26774377 7 50 theme gastric 1070:1076 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 50 theme gastric 1070:1076 arg1 acidity 1090:1096 gastric juice total acidity 1070:1096 gastric juice total acidity 1070:1096 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 3 51 theme homogeneous 293:303 arg1 component 305:313 a homogeneous component 291:313 a homogeneous component with an average molecular weight of 3.22 × 10(4)Da 291:364 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 3 51 theme homogeneous 293:303 arg1 LEP-2a 263:268 LEP-2a 263:268 LEP-2a 263:268 LEP-2a was identified to be a homogeneous component with an average molecular weight of 3.22 × 10(4)Da. 26774377 2 52 theme column 198:203 arg1 chromatography 205:218 DEAE-cellulose 52 column chromatography 180:218 DEAE-cellulose 52 column chromatography 180:218 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 7 53 theme juice 1056:1060 arg1 volume 1062:1067 gastric juice volume 1048:1067 gastric juice volume 1048:1067 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 53 theme juice 1056:1060 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 5 54 attach linked 728:733 arg3 C6 738:739 C6 738:739 C6 of the backbone chain 738:761 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 5 54 attach linked 728:733 arg3 linkage 783:789 β-1,6-glycosidic linkage 766:789 β-1,6-glycosidic linkage 766:789 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 5 54 attach linked 728:733 arg2 linkage 709:715 β-1,3-glycosidic linkage 692:715 β-1,3-glycosidic linkage 692:715 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 0 55 from sp 107:108 arg1 expolysaccharide 77:92 an expolysaccharide 74:92 an expolysaccharide from Lachnum sp 74:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 0 55 from sp 107:108 arg1 Structure 0:8 Structure 0:8 Structure 0:8 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 0 55 from sp 107:108 arg1 effects 25:31 preventive effects 14:31 preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp 14:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 1 56 theme Lachnum 146:152 arg1 sp 154:155 Lachnum sp 146:155 Lachnum sp 146:155 An extracellular polysaccharide of Lachnum sp. 26774377 2 57 theme DEAE-cellulose 180:193 arg1 chromatography 205:218 DEAE-cellulose 52 column chromatography 180:218 DEAE-cellulose 52 column chromatography 180:218 (LEP) was purified by DEAE-cellulose 52 column chromatography and Sepharose CL-6B column chromatography. 26774377 1 58 theme sp 154:155 arg1 polysaccharide 128:141 An extracellular polysaccharide 111:141 An extracellular polysaccharide of Lachnum sp 111:155 An extracellular polysaccharide of Lachnum sp. 26774377 9 59 theme ethanol-induced 1302:1316 arg1 ulcer 1326:1330 the ethanol-induced gastric ulcer 1298:1330 the ethanol-induced gastric ulcer 1298:1330 The above findings suggested that LEP-2a had a significant preventive effect against the ethanol-induced gastric ulcer. 26774377 0 60 from effects 25:31 arg1 sp 107:108 Lachnum sp 99:108 Lachnum sp 99:108 Structure and preventive effects against ethanol-induced gastric ulcer of an expolysaccharide from Lachnum sp. 26774377 7 61 theme juice 1078:1082 arg1 areas 1024:1028 the ulcer bleeding areas 1005:1028 the ulcer bleeding areas 1005:1028 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 7 61 theme juice 1078:1082 arg1 acidity 1090:1096 gastric juice total acidity 1070:1096 gastric juice total acidity 1070:1096 Results showed that LEP-2a significantly decreased the ulcer bleeding areas, pepsin activity, gastric juice volume, gastric juice total acidity and the malondialdehyde (MDA) content in serum. 26774377 5 62 theme chain 757:761 arg1 C6 738:739 C6 738:739 C6 of the backbone chain 738:761 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 26774377 5 63 theme β-1,3-glycosidic 692:707 arg1 linkage 709:715 β-1,3-glycosidic linkage 692:715 β-1,3-glycosidic linkage 692:715 Results indicated that LEP-2a was a (1→3)-,(1→6)-β-D-Glcp, whose branch chain was consist of two d-glucopyranosyl residues linked by β-1,3-glycosidic linkage, which was linked at C6 of the backbone chain by β-1,6-glycosidic linkage. 28402650 0 0 from Approach 25:32 arg1 Exosomes 92:99 Urinary Exosomes 84:99 Urinary Exosomes 84:99 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes. 28402650 8 1 theme liquid 1063:1068 arg1 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 8 1 theme liquid 1063:1068 arg1 spectrometry 1097:1108 liquid chromatography/tandem mass spectrometry 1063:1108 liquid chromatography/tandem mass spectrometry (LC/MS-MS) 1063:1119 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 4 2 contain have 575:578 arg2 size 591:594 an average size 580:594 an average size of 146 nm 580:604 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 4 2 contain have 575:578 arg1 exosomes 566:573 The isolated exosomes 553:573 The isolated exosomes 553:573 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 4 2 contain have 575:578 arg2 shape 622:626 a spherical shape 610:626 a spherical shape 610:626 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 12 3 theme unusual 1858:1864 arg1 structures 1866:1875 some unusual structures 1853:1875 some unusual structures 1853:1875 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 2 4 with biomarkers 338:347 arg1 results 369:375 some promising results 354:375 some promising results 354:375 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 9 5 theme graphitized 1301:1311 arg1 carbon 1313:1318 porous graphitized carbon 1294:1318 porous graphitized carbon (PGC) packed in long capillaries 1294:1351 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 9 5 theme graphitized 1301:1311 arg1 PGC 1321:1323 PGC 1321:1323 PGC 1321:1323 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 9 6 theme porous 1294:1299 arg1 carbon 1313:1318 porous graphitized carbon 1294:1318 porous graphitized carbon (PGC) packed in long capillaries 1294:1351 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 9 6 theme porous 1294:1299 arg1 PGC 1321:1323 PGC 1321:1323 PGC 1321:1323 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 7 7 theme relative 962:969 arg1 abundances 971:980 their relative abundances 956:980 their relative abundances 956:980 Paucimannosidic, high-mannose, and complex type glycans were identified and their relative abundances were determined. 28402650 9 8 theme long 1336:1339 arg1 capillaries 1341:1351 long capillaries 1336:1351 long capillaries 1336:1351 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 4 9 theme average 583:589 arg1 size 591:594 an average size 580:594 an average size of 146 nm 580:604 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 8 10 theme mass 1092:1095 arg1 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 8 10 theme mass 1092:1095 arg1 spectrometry 1097:1108 liquid chromatography/tandem mass spectrometry 1063:1108 liquid chromatography/tandem mass spectrometry (LC/MS-MS) 1063:1119 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 2 11 theme cancer 331:336 arg1 biomarkers 338:347 cancer biomarkers 331:347 cancer biomarkers with some promising results 331:375 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 4 12 theme dynamic 646:652 arg1 scattering 660:669 dynamic light scattering 646:669 dynamic light scattering 646:669 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 11 13 theme sulfated 1695:1702 arg1 glycans 1704:1710 sulfated glycans 1695:1710 sulfated glycans 1695:1710 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 1 14 dep bilayers 159:166 arg1 both 210:213 both 210:213 both 210:213 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 11 15 theme large 1581:1585 arg1 glycans 1587:1593 numerous large glycans 1572:1593 numerous large glycans 1572:1593 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 11 16 theme glycans 1704:1710 arg1 detection 1682:1690 detection 1682:1690 detection of sulfated glycans 1682:1710 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 3 17 theme urine 492:496 arg1 samples 498:504 normal urine samples 485:504 normal urine samples 485:504 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 11 18 theme C18 1549:1551 arg1 cartridge 1553:1561 a C18 cartridge 1547:1561 a C18 cartridge 1547:1561 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 11 19 dep revealed 1563:1570 arg1 whereas 1596:1602 whereas 1596:1602 whereas 1596:1602 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 10 20 theme multiantennary 1402:1415 arg1 N-glycans 1417:1425 minor multiantennary N-glycans 1396:1425 minor multiantennary N-glycans 1396:1425 Using microfractionation before LC/MS-MS, minor multiantennary N-glycans were preconcentrated as based on hydrophobicity or charge. 28402650 2 21 theme glycoconjugate 433:446 arg1 content 448:454 their glycoconjugate content 427:454 their glycoconjugate content 427:454 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 0 22 theme Analytical 14:23 arg1 Approach 25:32 Comprehensive Analytical Approach 0:32 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes 0:99 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes. 28402650 4 23 theme isolated 557:564 arg1 exosomes 566:573 The isolated exosomes 553:573 The isolated exosomes 553:573 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 1 24 with vesicles 139:146 arg1 bilayers 159:166 lipid bilayers 153:166 lipid bilayers encapsulating nucleic acids and proteins 153:207 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 11 25 theme glycans 1536:1542 arg1 Preconcentration 1486:1501 Preconcentration 1486:1501 Preconcentration of the reduced and permethylated glycans on a C18 cartridge 1486:1561 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 8 26 theme detailed 1004:1011 arg1 structures 1013:1022 Some detailed structures 999:1022 Some detailed structures of these glycans 999:1039 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 10 27 theme minor 1396:1400 arg1 N-glycans 1417:1425 minor multiantennary N-glycans 1396:1425 minor multiantennary N-glycans 1396:1425 Using microfractionation before LC/MS-MS, minor multiantennary N-glycans were preconcentrated as based on hydrophobicity or charge. 28402650 0 28 theme Comprehensive 0:12 arg1 Approach 25:32 Comprehensive Analytical Approach 0:32 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes 0:99 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes. 28402650 4 29 theme spherical 612:620 arg1 shape 622:626 a spherical shape 610:626 a spherical shape 610:626 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 6 30 theme mass 861:864 arg1 spectrometry 866:877 MALDI mass spectrometry 855:877 MALDI mass spectrometry 855:877 After being reduced and permethylated, N-glycans were measured by MALDI mass spectrometry. 28402650 7 31 theme type 923:926 arg1 glycans 928:934 complex type glycans 915:934 complex type glycans 915:934 Paucimannosidic, high-mannose, and complex type glycans were identified and their relative abundances were determined. 28402650 3 32 theme normal 485:490 arg1 samples 498:504 normal urine samples 485:504 normal urine samples 485:504 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 6 33 theme MALDI 855:859 arg1 spectrometry 866:877 MALDI mass spectrometry 855:877 MALDI mass spectrometry 855:877 After being reduced and permethylated, N-glycans were measured by MALDI mass spectrometry. 28402650 1 34 theme extracellular 115:127 arg1 vesicles 139:146 extracellular nanosized vesicles 115:146 extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation 115:244 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 1 34 theme extracellular 115:127 arg1 Exosomes 102:109 Exosomes 102:109 Exosomes 102:109 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 5 35 attach released 752:759 arg2 N-Glycans 723:731 N-Glycans 723:731 N-Glycans 723:731 N-Glycans were enzymatically released from the isolated vesicles. 28402650 5 35 attach released 752:759 arg1 vesicles 779:786 the isolated vesicles 766:786 the isolated vesicles 766:786 N-Glycans were enzymatically released from the isolated vesicles. 28402650 3 36 theme differential 524:535 arg1 centrifugation 537:550 multistep differential centrifugation 514:550 multistep differential centrifugation 514:550 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 3 37 attach isolated 471:478 arg2 Exosomes 457:464 Exosomes 457:464 Exosomes 457:464 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 3 37 attach isolated 471:478 arg1 samples 498:504 normal urine samples 485:504 normal urine samples 485:504 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 1 38 theme nanosized 129:137 arg1 vesicles 139:146 extracellular nanosized vesicles 115:146 extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation 115:244 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 1 38 theme nanosized 129:137 arg1 Exosomes 102:109 Exosomes 102:109 Exosomes 102:109 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 4 39 theme electron 688:695 arg1 microscopy 697:706 transmission electron microscopy 675:706 transmission electron microscopy 675:706 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 11 40 theme the 1506:1508 arg1 Preconcentration 1486:1501 Preconcentration 1486:1501 Preconcentration of the reduced and permethylated glycans on a C18 cartridge 1486:1561 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 7 41 theme complex 915:921 arg1 glycans 928:934 complex type glycans 915:934 complex type glycans 915:934 Paucimannosidic, high-mannose, and complex type glycans were identified and their relative abundances were determined. 28402650 0 42 theme Glycomic 41:48 arg1 Characterization 50:65 Glycomic Characterization 41:65 Glycomic Characterization 41:65 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes. 28402650 12 43 theme sample 1756:1761 arg1 aliquot 1763:1769 the original sample aliquot 1743:1769 the original sample aliquot 1743:1769 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 12 44 attach released 1798:1805 arg1 exosomes 1820:1827 urinary exosomes 1812:1827 urinary exosomes 1812:1827 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 12 44 attach released 1798:1805 arg2 O-glycans 1772:1780 O-glycans 1772:1780 O-glycans 1772:1780 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 2 45 theme nucleic 262:268 arg1 acids 270:274 exosomal nucleic acids 253:274 exosomal nucleic acids 253:274 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 3 46 theme multistep 514:522 arg1 centrifugation 537:550 multistep differential centrifugation 514:550 multistep differential centrifugation 514:550 Exosomes were isolated from normal urine samples through multistep differential centrifugation. 28402650 12 47 theme original 1747:1754 arg1 aliquot 1763:1769 the original sample aliquot 1743:1769 the original sample aliquot 1743:1769 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 12 48 theme urinary 1812:1818 arg1 exosomes 1820:1827 urinary exosomes 1812:1827 urinary exosomes 1812:1827 After removing N-glycans from the original sample aliquot, O-glycans were chemically released from urinary exosomes and profiled, revealing some unusual structures. 28402650 2 49 theme exosomal 253:260 arg1 acids 270:274 exosomal nucleic acids 253:274 exosomal nucleic acids 253:274 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 2 50 theme little 378:383 arg1 information 385:395 little information 378:395 little information 378:395 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 9 51 theme reduced 1126:1132 arg1 N-glycans 1134:1142 The reduced N-glycans 1122:1142 The reduced N-glycans 1122:1142 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 11 52 theme numerous 1572:1579 arg1 glycans 1587:1593 numerous large glycans 1572:1593 numerous large glycans 1572:1593 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 4 53 theme nm 603:604 arg1 shape 622:626 a spherical shape 610:626 a spherical shape 610:626 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 4 53 theme nm 603:604 arg1 size 591:594 an average size 580:594 an average size of 146 nm 580:604 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 11 54 theme permethylated 1522:1534 arg1 glycans 1536:1542 permethylated glycans 1522:1542 permethylated glycans 1522:1542 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 8 55 theme glycans 1033:1039 arg1 structures 1013:1022 Some detailed structures 999:1022 Some detailed structures of these glycans 999:1039 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 1 56 theme lipid 153:157 arg1 bilayers 159:166 lipid bilayers 153:166 lipid bilayers encapsulating nucleic acids and proteins 153:207 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28402650 11 57 theme ion-exchange 1646:1657 arg1 cartridges 1659:1668 ion-exchange cartridges 1646:1668 ion-exchange cartridges 1646:1668 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 5 58 theme isolated 770:777 arg1 vesicles 779:786 the isolated vesicles 766:786 the isolated vesicles 766:786 N-Glycans were enzymatically released from the isolated vesicles. 28402650 2 59 theme promising 359:367 arg1 results 369:375 some promising results 354:375 some promising results 354:375 While exosomal nucleic acids and proteins have previously been explored to identify cancer biomarkers with some promising results, little information has been available concerning their glycoconjugate content. 28402650 11 60 theme reduced 1625:1631 arg1 N-glycans 1633:1641 the reduced N-glycans 1621:1641 the reduced N-glycans 1621:1641 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 4 61 theme light 654:658 arg1 scattering 660:669 dynamic light scattering 646:669 dynamic light scattering 646:669 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 0 62 theme Urinary 84:90 arg1 Exosomes 92:99 Urinary Exosomes 84:99 Urinary Exosomes 84:99 Comprehensive Analytical Approach toward Glycomic Characterization and Profiling in Urinary Exosomes. 28402650 9 63 theme isomeric 1271:1278 arg1 separation 1280:1289 isomeric separation 1271:1289 isomeric separation 1271:1289 The reduced N-glycans, without being permethylated, were also separated and analyzed by LC/MS-MS, and their structures were further detailed through isomeric separation on porous graphitized carbon (PGC) packed in long capillaries. 28402650 11 64 theme N-glycans 1633:1641 arg1 fractionation 1604:1616 fractionation 1604:1616 fractionation of the reduced N-glycans by ion-exchange cartridges 1604:1668 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 4 65 theme transmission 675:686 arg1 microscopy 697:706 transmission electron microscopy 675:706 transmission electron microscopy 675:706 The isolated exosomes have an average size of 146 nm and a spherical shape, as determined by dynamic light scattering and transmission electron microscopy, respectively. 28402650 11 66 from Preconcentration 1486:1501 arg1 cartridge 1553:1561 a C18 cartridge 1547:1561 a C18 cartridge 1547:1561 Preconcentration of the reduced and permethylated glycans on a C18 cartridge revealed numerous large glycans, whereas fractionation of the reduced N-glycans by ion-exchange cartridges facilitated detection of sulfated glycans. 28402650 8 67 theme chromatography/tandem 1070:1090 arg1 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 LC/MS-MS 1111:1118 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 8 67 theme chromatography/tandem 1070:1090 arg1 spectrometry 1097:1108 liquid chromatography/tandem mass spectrometry 1063:1108 liquid chromatography/tandem mass spectrometry (LC/MS-MS) 1063:1119 Some detailed structures of these glycans were revealed through liquid chromatography/tandem mass spectrometry (LC/MS-MS). 28402650 1 68 theme nucleic 182:188 arg1 acids 190:194 nucleic acids 182:194 nucleic acids 182:194 Exosomes are extracellular nanosized vesicles with lipid bilayers encapsulating nucleic acids and proteins, both with and without glycosylation. 28556908 0 0 theme anionic 87:93 arg1 N-glycans 109:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 6 1 theme N-glycan 1623:1630 arg1 structures 1632:1641 neutral and anionic N-glycan structures 1603:1641 neutral and anionic N-glycan structures 1603:1641 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 3 2 theme unusual 887:893 arg1 extensions 914:923 unusual oligo-α1,2-mannose extensions 887:923 unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications 887:979 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 2 3 theme anionic 641:647 arg1 N-glycome 649:657 the anionic N-glycome 637:657 the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens 637:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 4 theme cellular 163:170 arg1 discoideum 198:207 the cellular slime mould Dictyostelium discoideum 159:207 the cellular slime mould Dictyostelium discoideum 159:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 4 5 theme isobaric 1058:1065 arg1 glycans 1067:1073 isobaric glycans 1058:1073 isobaric glycans differing in the presence of sulphate or phosphate 1058:1124 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 3 6 theme HIAX 822:825 arg1 chromatography 827:840 HIAX chromatography 822:840 HIAX chromatography 822:840 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 0 7 theme N-glycans 109:117 arg1 separation 43:52 separation 43:52 separation of multiply modified neutral and anionic Dictyostelium N-glycans 43:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 5 8 theme enzymes 1466:1472 arg1 function 1363:1370 a similar function 1353:1370 a similar function 1353:1370 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 8 theme enzymes 1466:1472 arg1 modification 1440:1451 the mannose-6-phosphate modification 1416:1451 the mannose-6-phosphate modification of lysosomal enzymes in mammals 1416:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 1 9 theme anionic 440:446 arg1 modifications 448:460 anionic modifications 440:460 anionic modifications 440:460 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 3 10 theme N-glycans 872:880 arg1 identification 854:867 the identification 850:867 the identification of N-glycans 850:880 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 5 11 theme biosynthetic 1226:1237 arg1 intermediates 1239:1251 biosynthetic intermediates 1226:1251 biosynthetic intermediates 1226:1251 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 0 12 theme Dictyostelium 95:107 arg1 N-glycans 109:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 3 13 with Prefractionation 800:815 arg1 chromatography 827:840 HIAX chromatography 822:840 HIAX chromatography 822:840 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 1 14 theme modifications 448:460 arg1 occurrence 426:435 the occurrence 422:435 the occurrence of anionic modifications, which also present an analytical challenge 422:504 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 5 15 theme similar 1355:1361 arg1 function 1363:1370 a similar function 1353:1370 a similar function 1353:1370 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 15 theme similar 1355:1361 arg1 modification 1440:1451 the mannose-6-phosphate modification 1416:1451 the mannose-6-phosphate modification of lysosomal enzymes in mammals 1416:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 16 theme intracellular 1375:1387 arg1 targeting 1389:1397 intracellular targeting 1375:1397 intracellular targeting of hydrolases 1375:1411 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 4 17 contain carrying 1172:1179 arg2 GlcNAc-phosphodiesters 1181:1202 GlcNAc-phosphodiesters 1181:1202 GlcNAc-phosphodiesters 1181:1202 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 4 17 contain carrying 1172:1179 arg1 those 1166:1170 those 1166:1170 those 1166:1170 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 6 18 theme anionic 1615:1621 arg1 structures 1632:1641 neutral and anionic N-glycan structures 1603:1641 neutral and anionic N-glycan structures 1603:1641 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 3 19 dep four 954:957 arg1 to 951:952 to 951:952 to 951:952 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 1 20 theme slime 172:176 arg1 discoideum 198:207 the cellular slime mould Dictyostelium discoideum 159:207 the cellular slime mould Dictyostelium discoideum 159:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 1 21 theme N-glycomes 364:373 arg1 complexity 346:355 The complexity 342:355 The complexity of the N-glycomes of even glycosylation mutants 342:403 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 5 22 theme lysosomal 1456:1464 arg1 enzymes 1466:1472 lysosomal enzymes 1456:1472 lysosomal enzymes in mammals 1456:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 4 23 theme treatment 1018:1026 arg1 use 993:995 the use 989:995 the use of hydrofluoric acid treatment 989:1026 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 2 24 with combination 593:603 arg1 MS/MS 620:624 MALDI-TOF MS/MS 610:624 MALDI-TOF MS/MS 610:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 25 theme mould 178:182 arg1 discoideum 198:207 the cellular slime mould Dictyostelium discoideum 159:207 the cellular slime mould Dictyostelium discoideum 159:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 0 26 theme interaction 12:22 arg1 exchange 30:37 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans 0:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 0:118 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 2 27 theme MALDI-TOF 610:618 arg1 MS/MS 620:624 MALDI-TOF MS/MS 610:624 MALDI-TOF MS/MS 610:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 28 theme M31 666:668 arg1 strain 677:682 the M31 (modA) strain 662:682 the M31 (modA) strain 662:682 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 6 29 from approach 1551:1558 arg1 conclusion 1489:1498 conclusion 1489:1498 conclusion 1489:1498 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 4 30 theme intersected 1129:1139 arg1 structures 1141:1150 intersected structures 1129:1150 intersected structures 1129:1150 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 1 31 theme Dictyostelium 184:196 arg1 discoideum 198:207 the cellular slime mould Dictyostelium discoideum 159:207 the cellular slime mould Dictyostelium discoideum 159:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 0 32 theme Hydrophilic 0:10 arg1 exchange 30:37 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans 0:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 0:118 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 2 33 from HPLC 585:588 arg1 combination 593:603 combination 593:603 combination with MALDI-TOF MS/MS 593:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 6 34 with combination 1509:1519 arg1 MS 1526:1527 MS 1526:1527 MS 1526:1527 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 6 35 theme structures 1632:1641 arg1 separation 1574:1583 fine separation 1569:1583 fine separation 1569:1583 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 6 35 theme structures 1632:1641 arg1 definition 1589:1598 definition 1589:1598 definition of neutral and anionic N-glycan structures 1589:1641 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 5 36 contain have 1348:1351 arg2 function 1363:1370 a similar function 1353:1370 a similar function 1353:1370 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 36 contain have 1348:1351 arg2 modification 1440:1451 the mannose-6-phosphate modification 1416:1451 the mannose-6-phosphate modification of lysosomal enzymes in mammals 1416:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 36 contain have 1348:1351 arg1 epitope 1329:1335 the methylphosphorylated mannose epitope 1296:1335 the methylphosphorylated mannose epitope 1296:1335 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 2 37 theme most 760:763 arg1 eukaryotes 765:774 most eukaryotes 760:774 most eukaryotes including Homo sapiens 760:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 37 theme most 760:763 arg1 sapiens 791:797 Homo sapiens 786:797 Homo sapiens 786:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 38 theme discoideum 198:207 arg1 N-glycans 146:154 the N-glycans 142:154 the N-glycans of the cellular slime mould Dictyostelium discoideum 142:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 2 39 theme interaction 551:561 arg1 HPLC 585:588 hydrophilic interaction anion exchange (HIAX) HPLC 539:588 hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS 539:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 6 40 theme fine 1569:1572 arg1 separation 1574:1583 fine separation 1569:1583 fine separation 1569:1583 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 5 41 from modification 1440:1451 arg1 mammals 1477:1483 mammals 1477:1483 mammals 1477:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 0 42 theme anion 24:28 arg1 exchange 30:37 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans 0:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 0:118 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 1 43 theme radiolabeled 285:296 arg1 glycopeptides 298:310 radiolabeled glycopeptides 285:310 radiolabeled glycopeptides 285:310 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 2 44 theme hydrophilic 539:549 arg1 HPLC 585:588 hydrophilic interaction anion exchange (HIAX) HPLC 539:588 hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS 539:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 45 theme even 378:381 arg1 mutants 397:403 even glycosylation mutants 378:403 even glycosylation mutants 378:403 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 6 46 theme sensitive 1541:1549 arg1 HIAX 1501:1504 HIAX 1501:1504 HIAX in combination with MS 1501:1527 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 6 46 theme sensitive 1541:1549 arg1 approach 1551:1558 a highly sensitive approach 1532:1558 a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures 1532:1641 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 1 47 theme glycopeptides 298:310 arg1 examination 270:280 examination 270:280 examination of radiolabeled glycopeptides 270:310 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 1 48 theme glycosylation 383:395 arg1 mutants 397:403 even glycosylation mutants 378:403 even glycosylation mutants 378:403 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 5 49 theme mannose-6-phosphate 1420:1438 arg1 function 1363:1370 a similar function 1353:1370 a similar function 1353:1370 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 49 theme mannose-6-phosphate 1420:1438 arg1 modification 1440:1451 the mannose-6-phosphate modification 1416:1451 the mannose-6-phosphate modification of lysosomal enzymes in mammals 1416:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 4 50 theme sulphate 1104:1111 arg1 presence 1092:1099 the presence 1088:1099 the presence of sulphate or phosphate 1088:1124 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 1 51 theme mutants 397:403 arg1 N-glycomes 364:373 the N-glycomes 360:373 the N-glycomes of even glycosylation mutants 360:403 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 6 52 theme neutral 1603:1609 arg1 structures 1632:1641 neutral and anionic N-glycan structures 1603:1641 neutral and anionic N-glycan structures 1603:1641 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 1 53 theme analytical 485:494 arg1 challenge 496:504 an analytical challenge 482:504 an analytical challenge 482:504 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 4 54 theme acid 1013:1016 arg1 treatment 1018:1026 hydrofluoric acid treatment 1000:1026 hydrofluoric acid treatment 1000:1026 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 4 55 theme phosphate 1116:1124 arg1 presence 1092:1099 the presence 1088:1099 the presence of sulphate or phosphate 1088:1124 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 2 56 theme HIAX 579:582 arg1 HPLC 585:588 hydrophilic interaction anion exchange (HIAX) HPLC 539:588 hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS 539:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 57 theme strain 677:682 arg1 N-glycome 649:657 the anionic N-glycome 637:657 the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens 637:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 58 theme unusual 124:130 arg1 nature 132:137 The unusual nature 120:137 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum 120:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 4 59 theme hydrofluoric 1000:1011 arg1 acid 1013:1016 hydrofluoric acid 1000:1016 hydrofluoric acid treatment 1000:1026 Due to the use of hydrofluoric acid treatment, we were able to discriminate isobaric glycans differing in the presence of sulphate or phosphate on intersected structures as opposed to those carrying GlcNAc-phosphodiesters. 28556908 1 60 gly glycopeptides 298:310 arg2 glycopeptides 298:310 radiolabeled glycopeptides 285:310 radiolabeled glycopeptides 285:310 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 5 61 from mammals 1477:1483 arg1 function 1363:1370 a similar function 1353:1370 a similar function 1353:1370 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 61 from mammals 1477:1483 arg1 modification 1440:1451 the mannose-6-phosphate modification 1416:1451 the mannose-6-phosphate modification of lysosomal enzymes in mammals 1416:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 62 theme methylphosphorylated 1300:1319 arg1 epitope 1329:1335 the methylphosphorylated mannose epitope 1296:1335 the methylphosphorylated mannose epitope 1296:1335 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 2 63 theme exchange 569:576 arg1 HPLC 585:588 hydrophilic interaction anion exchange (HIAX) HPLC 539:588 hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS 539:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 3 64 with extensions 914:923 arg1 modifications 967:979 up to four anionic modifications 948:979 up to four anionic modifications 948:979 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 2 65 theme modA 671:674 arg1 strain 677:682 the M31 (modA) strain 662:682 the M31 (modA) strain 662:682 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 66 theme Homo 786:789 arg1 sapiens 791:797 Homo sapiens 786:797 Homo sapiens 786:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 67 theme anion 563:567 arg1 HPLC 585:588 hydrophilic interaction anion exchange (HIAX) HPLC 539:588 hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS 539:624 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 3 68 theme anionic 959:965 arg1 modifications 967:979 up to four anionic modifications 948:979 up to four anionic modifications 948:979 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 2 69 theme reticulum 709:717 arg1 α-glucosidase 719:731 endoplasmic reticulum α-glucosidase II 697:734 endoplasmic reticulum α-glucosidase II 697:734 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 69 theme reticulum 709:717 arg1 enzyme 740:745 an enzyme 737:745 an enzyme conserved in most eukaryotes including Homo sapiens 737:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 5 70 from enzymes 1466:1472 arg1 mammals 1477:1483 mammals 1477:1483 mammals 1477:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 5 71 theme epitope 1329:1335 arg1 formation 1283:1291 formation 1283:1291 formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals 1283:1483 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 3 72 theme oligo-α1,2-mannose 895:912 arg1 extensions 914:923 unusual oligo-α1,2-mannose extensions 887:923 unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications 887:979 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 0 73 theme neutral 75:81 arg1 N-glycans 109:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 6 74 from HIAX 1501:1504 arg1 combination 1509:1519 combination 1509:1519 combination with MS 1509:1527 In conclusion, HIAX in combination with MS is a highly sensitive approach for both fine separation and definition of neutral and anionic N-glycan structures. 28556908 2 75 theme endoplasmic 697:707 arg1 α-glucosidase 719:731 endoplasmic reticulum α-glucosidase II 697:734 endoplasmic reticulum α-glucosidase II 697:734 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 2 75 theme endoplasmic 697:707 arg1 enzyme 740:745 an enzyme 737:745 an enzyme conserved in most eukaryotes including Homo sapiens 737:797 In this study, we have employed hydrophilic interaction anion exchange (HIAX) HPLC in combination with MALDI-TOF MS/MS to explore the anionic N-glycome of the M31 (modA) strain, which lacks endoplasmic reticulum α-glucosidase II, an enzyme conserved in most eukaryotes including Homo sapiens. 28556908 1 76 theme N-glycans 146:154 arg1 nature 132:137 The unusual nature 120:137 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum 120:207 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28556908 5 77 theme mannose 1321:1327 arg1 epitope 1329:1335 the methylphosphorylated mannose epitope 1296:1335 the methylphosphorylated mannose epitope 1296:1335 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 0 78 theme modified 66:73 arg1 N-glycans 109:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 multiply modified neutral and anionic Dictyostelium N-glycans 57:117 Hydrophilic interaction anion exchange for separation of multiply modified neutral and anionic Dictyostelium N-glycans. 28556908 5 79 theme hydrolases 1402:1411 arg1 targeting 1389:1397 intracellular targeting 1375:1397 intracellular targeting of hydrolases 1375:1411 The latter represent biosynthetic intermediates during the pathway leading to formation of the methylphosphorylated mannose epitope, which may have a similar function in intracellular targeting of hydrolases as the mannose-6-phosphate modification of lysosomal enzymes in mammals. 28556908 3 80 with others 936:941 arg1 modifications 967:979 up to four anionic modifications 948:979 up to four anionic modifications 948:979 Prefractionation with HIAX chromatography enabled the identification of N-glycans with unusual oligo-α1,2-mannose extensions as well as others with up to four anionic modifications. 28556908 1 81 theme studies 242:248 arg1 number 232:237 a number 230:237 a number of studies 230:248 The unusual nature of the N-glycans of the cellular slime mould Dictyostelium discoideum has been revealed by a number of studies, primarily based on examination of radiolabeled glycopeptides but more recently also by MS. The complexity of the N-glycomes of even glycosylation mutants is compounded by the occurrence of anionic modifications, which also present an analytical challenge. 28211926 2 0 theme access 448:453 arg1 glycoconjugates 455:469 access glycoconjugates 448:469 access glycoconjugates 448:469 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 2 0 theme access 448:453 arg1 glycopeptides 480:492 glycopeptides 480:492 glycopeptides 480:492 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 1 1 with reaction 294:301 arg1 acid 319:322 thioacetic acid 308:322 thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate 308:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 2 theme sugar 245:249 arg1 groups 262:267 any sugar projecting groups 241:267 any sugar projecting groups 241:267 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 2 3 used used 440:443 arg2 thiols 402:407 Glycosyl thiols 393:407 Glycosyl thiols produced by this method 393:431 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 2 4 gly glycopeptides 480:492 arg2 glycopeptides 480:492 glycopeptides 480:492 glycopeptides 480:492 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 1 5 theme projecting 251:260 arg1 groups 262:267 any sugar projecting groups 241:267 any sugar projecting groups 241:267 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 6 theme thiols 44:49 arg1 synthesis 22:30 group free synthesis 11:30 group free synthesis of glycosyl thiols 11:49 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 1 7 theme base 343:346 arg1 presence 331:338 the presence 327:338 the presence of base, and hydrolysis of the anomeric thioacetate 327:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 8 theme group 11:15 arg1 synthesis 22:30 group free synthesis 11:30 group free synthesis of glycosyl thiols 11:49 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 2 9 theme thiol-ene 508:516 arg1 reaction 524:531 the thiol-ene click reaction 504:531 the thiol-ene click reaction 504:531 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 1 10 theme corresponding 182:194 arg1 sugars 205:210 the corresponding reducing sugars 178:210 the corresponding reducing sugars in water without recourse to any sugar projecting groups 178:267 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 11 theme free 17:20 arg1 synthesis 22:30 group free synthesis 11:30 group free synthesis of glycosyl thiols 11:49 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 1 12 theme reducing 196:203 arg1 sugars 205:210 the corresponding reducing sugars 178:210 the corresponding reducing sugars in water without recourse to any sugar projecting groups 178:267 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 13 theme glycosyl 35:42 arg1 thiols 44:49 glycosyl thiols 35:49 glycosyl thiols 35:49 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 0 14 theme glycoconjugates 124:138 arg1 production 101:110 the production 97:110 the production of N-glycan glycoconjugates 97:138 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 1 15 theme hydrolysis 353:362 arg1 presence 331:338 the presence 327:338 the presence of base, and hydrolysis of the anomeric thioacetate 327:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 16 theme N-glycan 115:122 arg1 glycoconjugates 124:138 N-glycan glycoconjugates 115:138 N-glycan glycoconjugates 115:138 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 0 17 from sugars 65:70 arg1 water 75:79 water 75:79 water 75:79 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 1 18 from sugars 205:210 arg1 water 215:219 water 215:219 water without recourse to any sugar projecting groups 215:267 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 19 theme mediated 285:292 arg1 reaction 294:301 a DMC mediated reaction 279:301 a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate 279:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 20 from acid 319:322 arg1 presence 331:338 the presence 327:338 the presence of base, and hydrolysis of the anomeric thioacetate 327:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 21 theme reaction 294:301 arg1 way 272:274 way 272:274 way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate 272:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 22 theme anomeric 371:378 arg1 thioacetate 380:390 the anomeric thioacetate 367:390 the anomeric thioacetate 367:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 2 23 theme reaction 524:531 arg1 use 497:499 use 497:499 use of the thiol-ene click reaction 497:531 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 1 24 theme thioacetate 380:390 arg1 base 343:346 base 343:346 base 343:346 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 1 24 theme thioacetate 380:390 arg1 hydrolysis 353:362 hydrolysis 353:362 hydrolysis of the anomeric thioacetate 353:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 2 25 theme click 518:522 arg1 reaction 524:531 the thiol-ene click reaction 504:531 the thiol-ene click reaction 504:531 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 2 26 theme Glycosyl 393:400 arg1 thiols 402:407 Glycosyl thiols 393:407 Glycosyl thiols produced by this method 393:431 Glycosyl thiols produced by this method may be used to access glycoconjugates, such as glycopeptides by use of the thiol-ene click reaction. 28211926 1 27 theme thioacetic 308:317 arg1 acid 319:322 thioacetic acid 308:322 thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate 308:390 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 28211926 0 28 dep Protecting 0:9 arg1 application 82:92 application 82:92 application to the production of N-glycan glycoconjugates 82:138 Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates. 28211926 1 29 theme Glycosyl 141:148 arg1 thiols 150:155 Glycosyl thiols 141:155 Glycosyl thiols 141:155 Glycosyl thiols may be accessed from the corresponding reducing sugars in water without recourse to any sugar projecting groups by way of a DMC mediated reaction with thioacetic acid in the presence of base, and hydrolysis of the anomeric thioacetate. 27863303 3 0 from modification 495:506 arg1 skeleton 547:554 the dihydroresveratrol skeleton 524:554 the dihydroresveratrol skeleton 524:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 2 1 theme well-known 429:438 arg1 8-methoxypsoralen 408:424 8-methoxypsoralen 408:424 8-methoxypsoralen 408:424 Both analogues stimulated melanogenesis with efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator. 27863303 2 1 theme well-known 429:438 arg1 activator 454:462 a well-known melanogenesis activator 427:462 a well-known melanogenesis activator 427:462 Both analogues stimulated melanogenesis with efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator. 27863303 3 2 from groups 634:639 arg1 moieties 654:661 the sugar moieties 644:661 the sugar moieties 644:661 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 1 3 theme key 320:322 arg1 cellobioside 106:117 Dihydroresveratrol cellobioside 87:117 Dihydroresveratrol cellobioside 87:117 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 3 theme key 320:322 arg1 step 324:327 the key step 316:327 the key step 316:327 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 3 theme key 320:322 arg1 xylobioside 123:133 xylobioside 123:133 xylobioside 123:133 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 4 theme Dihydroresveratrol 87:104 arg1 cellobioside 106:117 Dihydroresveratrol cellobioside 87:117 Dihydroresveratrol cellobioside 87:117 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 4 theme Dihydroresveratrol 87:104 arg1 step 324:327 the key step 316:327 the key step 316:327 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 4 theme Dihydroresveratrol 87:104 arg1 xylobioside 123:133 xylobioside 123:133 xylobioside 123:133 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 3 5 theme hydroxymethyl 620:632 arg1 groups 634:639 hydroxymethyl groups 620:639 hydroxymethyl groups in the sugar moieties 620:661 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 1 6 theme agent 234:238 arg1 glucoside 259:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 7 theme occurring 198:206 arg1 glucoside 259:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 1 8 theme melanogenesis-controlling 208:232 arg1 glucoside 259:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 3 9 theme 4'-OH 515:519 arg1 modification 495:506 diglycosyl modification 484:506 diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton 484:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 2 10 theme comparable 386:395 arg1 efficacies 375:384 efficacies 375:384 efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator 375:462 Both analogues stimulated melanogenesis with efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator. 27863303 1 11 theme dihydroresveratrol 240:257 arg1 glucoside 259:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside 184:267 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 3 12 theme sugar 648:652 arg1 moieties 654:661 the sugar moieties 644:661 the sugar moieties 644:661 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 0 13 theme effective 51:59 arg1 activators 75:84 effective melanogenesis activators 51:84 effective melanogenesis activators 51:84 Dihydroresveratrol cellobioside and xylobioside as effective melanogenesis activators. 27863303 3 14 from skeleton 547:554 arg1 modification 495:506 diglycosyl modification 484:506 diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton 484:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 3 15 theme diglycosyl 484:493 arg1 modification 495:506 diglycosyl modification 484:506 diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton 484:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 3 16 theme melanogenesis 583:595 arg1 activation 569:578 the activation 565:578 the activation of melanogenesis 565:595 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 0 17 theme melanogenesis 61:73 arg1 activators 75:84 effective melanogenesis activators 51:84 effective melanogenesis activators 51:84 Dihydroresveratrol cellobioside and xylobioside as effective melanogenesis activators. 27863303 3 18 from 4'-OH 515:519 arg1 skeleton 547:554 the dihydroresveratrol skeleton 524:554 the dihydroresveratrol skeleton 524:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 1 19 theme Schmidt 291:297 arg1 glycosylation 299:311 Schmidt glycosylation 291:311 Schmidt glycosylation 291:311 Dihydroresveratrol cellobioside and xylobioside, whose structures were designed based on that of the naturally occurring melanogenesis-controlling agent dihydroresveratrol glucoside, were synthesized via Schmidt glycosylation as the key step. 27863303 3 20 theme dihydroresveratrol 528:545 arg1 skeleton 547:554 the dihydroresveratrol skeleton 524:554 the dihydroresveratrol skeleton 524:554 This suggests that diglycosyl modification of the 4'-OH on the dihydroresveratrol skeleton leads to the activation of melanogenesis, both with and without hydroxymethyl groups in the sugar moieties. 27863303 2 21 theme melanogenesis 440:452 arg1 8-methoxypsoralen 408:424 8-methoxypsoralen 408:424 8-methoxypsoralen 408:424 Both analogues stimulated melanogenesis with efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator. 27863303 2 21 theme melanogenesis 440:452 arg1 activator 454:462 a well-known melanogenesis activator 427:462 a well-known melanogenesis activator 427:462 Both analogues stimulated melanogenesis with efficacies comparable to that of 8-methoxypsoralen, a well-known melanogenesis activator. 28816453 5 0 theme relevant 806:813 arg1 trisaccharides 815:828 all relevant trisaccharides 802:828 all relevant trisaccharides 802:828 Next, we systematically study the effect of neighboring residues, by extensively simulating all relevant trisaccharides and one tetrasaccharide. 28816453 1 1 theme three-dimensional 165:181 arg1 data 194:197 relatively little three-dimensional structural data 147:197 relatively little three-dimensional structural data 147:197 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 7 2 theme small 958:962 arg1 number 964:969 a small number 956:969 a small number of exceptions 956:983 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 8 3 from O-glycans 1178:1186 arg1 glycoproteins 1191:1203 glycoproteins 1191:1203 glycoproteins 1191:1203 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 1 4 theme structural 183:192 arg1 data 194:197 relatively little three-dimensional structural data 147:197 relatively little three-dimensional structural data 147:197 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 5 5 theme residues 766:773 arg1 effect 744:749 the effect 740:749 the effect of neighboring residues 740:773 Next, we systematically study the effect of neighboring residues, by extensively simulating all relevant trisaccharides and one tetrasaccharide. 28816453 2 6 theme mammalian 304:312 arg1 oligosaccharides 314:329 mammalian oligosaccharides 304:329 mammalian oligosaccharides appearing in N- and O-glycosylated proteins 304:373 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 4 7 theme complete 626:633 arg1 library 650:656 the first complete conformational library 616:656 the first complete conformational library for the construction of N- and O-glycan structures 616:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 0 8 from Landscapes 67:76 arg1 Glycoproteins 36:48 Glycoproteins 36:48 Glycoproteins from Free-Energy Landscapes 36:76 Modeling of Oligosaccharides within Glycoproteins from Free-Energy Landscapes. 28816453 2 9 theme majority 292:299 arg1 flexibility 268:278 flexibility 268:278 flexibility 268:278 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 2 9 theme majority 292:299 arg1 structure 254:262 structure 254:262 structure 254:262 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 4 10 theme first 620:624 arg1 library 650:656 the first complete conformational library 616:656 the first complete conformational library for the construction of N- and O-glycan structures 616:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 1 11 gly glycoproteins 108:120 arg1 glycoproteins 108:120 glycoproteins 108:120 glycoproteins 108:120 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 8 12 theme relevant 1228:1235 arg1 examples 1237:1244 two relevant examples 1224:1244 two relevant examples 1224:1244 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 7 13 theme larger 1024:1029 arg1 structures 1031:1040 the larger structures 1020:1040 the larger structures 1020:1040 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 2 14 theme vast 287:290 arg1 majority 292:299 the vast majority 283:299 the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins 283:373 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 3 15 theme relevant 462:469 arg1 linkages 482:489 all relevant glycosidic linkages 458:489 all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling 458:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 3 16 theme conformational 417:430 arg1 landscapes 444:453 the conformational free-energy landscapes 413:453 the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling 413:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 6 17 from comparison 888:897 arg1 oligosaccharides 933:948 large oligosaccharides 927:948 large oligosaccharides 927:948 This allows for an unprecedented comparison of disaccharide linkages in large oligosaccharides. 28816453 4 18 theme conformational 635:648 arg1 library 650:656 the first complete conformational library 616:656 the first complete conformational library for the construction of N- and O-glycan structures 616:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 2 19 dep structure 254:262 arg1 the 250:252 the 250:252 the 250:252 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 8 20 theme several 1120:1126 arg1 approaches 1138:1147 several efficient approaches 1120:1147 several efficient approaches 1120:1147 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 4 21 theme O-glycan 689:696 arg1 construction 666:677 the construction 662:677 the construction of N- and O-glycan structures 662:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 8 22 theme complete 1162:1169 arg1 N- 1171:1172 complete N- 1162:1172 complete N- 1162:1172 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 2 23 dep N- 344:345 arg1 proteins 366:373 proteins 366:373 proteins 366:373 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 7 24 theme conformational 990:1003 arg1 preferences 1005:1015 the conformational preferences 986:1015 the conformational preferences in the larger structures 986:1040 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 7 24 theme conformational 990:1003 arg1 similar 1051:1057 similar 1051:1057 similar 1051:1057 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 2 25 theme oligosaccharides 314:329 arg1 majority 292:299 the vast majority 283:299 the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins 283:373 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 1 26 theme glycan 216:221 arg1 structures 223:232 glycan structures 216:232 glycan structures 216:232 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 0 27 theme Oligosaccharides 12:27 arg1 Modeling 0:7 Modeling 0:7 Modeling of Oligosaccharides within Glycoproteins from Free-Energy Landscapes. 0:77 Modeling of Oligosaccharides within Glycoproteins from Free-Energy Landscapes. 28816453 8 28 theme efficient 1128:1136 arg1 approaches 1138:1147 several efficient approaches 1120:1147 several efficient approaches 1120:1147 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 1 29 theme glycoproteins 108:120 arg1 abundance 95:103 the abundance 91:103 the abundance of glycoproteins in biological processes 91:144 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 7 30 from preferences 1005:1015 arg1 structures 1031:1040 the larger structures 1020:1040 the larger structures 1020:1040 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 7 31 from disaccharides 1069:1081 arg1 preferences 1005:1015 the conformational preferences 986:1015 the conformational preferences in the larger structures 986:1040 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 7 31 from disaccharides 1069:1081 arg1 similar 1051:1057 similar 1051:1057 similar 1051:1057 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 3 32 theme free-energy 432:442 arg1 landscapes 444:453 the conformational free-energy landscapes 413:453 the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling 413:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 6 33 theme large 927:931 arg1 oligosaccharides 933:948 large oligosaccharides 927:948 large oligosaccharides 927:948 This allows for an unprecedented comparison of disaccharide linkages in large oligosaccharides. 28816453 3 34 theme linkages 482:489 arg1 landscapes 444:453 the conformational free-energy landscapes 413:453 the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling 413:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 5 35 theme neighboring 754:764 arg1 residues 766:773 neighboring residues 754:773 neighboring residues 754:773 Next, we systematically study the effect of neighboring residues, by extensively simulating all relevant trisaccharides and one tetrasaccharide. 28816453 4 36 dep N- 682:683 arg1 structures 698:707 structures 698:707 structures 698:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 4 37 theme N- 682:683 arg1 construction 666:677 the construction 662:677 the construction of N- and O-glycan structures 662:707 To the best of our knowledge, this represents the first complete conformational library for the construction of N- and O-glycan structures. 28816453 3 38 theme subsequent 540:549 arg1 sampling 560:567 subsequent umbrella sampling 540:567 subsequent umbrella sampling 540:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 6 39 theme linkages 915:922 arg1 comparison 888:897 an unprecedented comparison 871:897 an unprecedented comparison of disaccharide linkages in large oligosaccharides 871:948 This allows for an unprecedented comparison of disaccharide linkages in large oligosaccharides. 28816453 8 40 gly glycoproteins 1191:1203 arg1 glycoproteins 1191:1203 glycoproteins 1191:1203 glycoproteins 1191:1203 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 0 41 theme Free-Energy 55:65 arg1 Landscapes 67:76 Free-Energy Landscapes 55:76 Free-Energy Landscapes 55:76 Modeling of Oligosaccharides within Glycoproteins from Free-Energy Landscapes. 28816453 7 42 from similar 1051:1057 arg1 disaccharides 1069:1081 the disaccharides 1065:1081 the disaccharides 1065:1081 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 6 43 theme disaccharide 902:913 arg1 linkages 915:922 disaccharide linkages 902:922 disaccharide linkages 902:922 This allows for an unprecedented comparison of disaccharide linkages in large oligosaccharides. 28816453 1 44 theme little 158:163 arg1 data 194:197 relatively little three-dimensional structural data 147:197 relatively little three-dimensional structural data 147:197 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 3 45 theme umbrella 551:558 arg1 sampling 560:567 subsequent umbrella sampling 540:567 subsequent umbrella sampling 540:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 1 46 theme biological 125:134 arg1 processes 136:144 biological processes 125:144 biological processes 125:144 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 7 47 theme exceptions 974:983 arg1 number 964:969 a small number 956:969 a small number of exceptions 956:983 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 7 48 with similar 1051:1057 arg1 number 964:969 a small number 956:969 a small number of exceptions 956:983 With a small number of exceptions, the conformational preferences in the larger structures are very similar as in the disaccharides. 28816453 3 49 theme local 508:512 arg1 simulations 524:534 local elevation simulations 508:534 local elevation simulations 508:534 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 3 50 theme elevation 514:522 arg1 simulations 524:534 local elevation simulations 508:534 local elevation simulations 508:534 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 3 51 theme glycosidic 471:480 arg1 linkages 482:489 all relevant glycosidic linkages 458:489 all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling 458:567 We report the conformational free-energy landscapes of all relevant glycosidic linkages as obtained from local elevation simulations and subsequent umbrella sampling. 28816453 6 52 theme unprecedented 874:886 arg1 comparison 888:897 an unprecedented comparison 871:897 an unprecedented comparison of disaccharide linkages in large oligosaccharides 871:948 This allows for an unprecedented comparison of disaccharide linkages in large oligosaccharides. 28816453 2 53 theme up 390:391 arg1 approach 393:400 a bottom up approach 381:400 a bottom up approach 381:400 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 28816453 1 54 from abundance 95:103 arg1 processes 136:144 biological processes 125:144 biological processes 125:144 In spite of the abundance of glycoproteins in biological processes, relatively little three-dimensional structural data is available for glycan structures. 28816453 8 55 from N- 1171:1172 arg1 glycoproteins 1191:1203 glycoproteins 1191:1203 glycoproteins 1191:1203 This, finally, allows us to suggest several efficient approaches to construct complete N- and O-glycans on glycoproteins, as exemplified on two relevant examples. 28816453 2 56 theme bottom 383:388 arg1 approach 393:400 a bottom up approach 381:400 a bottom up approach 381:400 Here, we study the structure and flexibility of the vast majority of mammalian oligosaccharides appearing in N- and O-glycosylated proteins using a bottom up approach. 25843303 5 0 theme glycosyl 840:847 arg1 family 859:864 glycosyl hydrolase family 840:864 glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively 840:1079 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 8 1 dep acceptors 1522:1530 arg1 acceptors 1522:1530 the disaccharide acceptors β-lactose, sucrose, and maltose 1505:1562 the disaccharide acceptors β-lactose, sucrose, and maltose 1505:1562 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 1 dep acceptors 1522:1530 arg1 sucrose 1543:1549 sucrose 1543:1549 sucrose 1543:1549 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 1 dep acceptors 1522:1530 arg1 maltose 1556:1562 maltose 1556:1562 maltose 1556:1562 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 1 dep acceptors 1522:1530 arg1 β-lactose 1532:1540 β-lactose 1532:1540 β-lactose 1532:1540 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 1 2 theme β-N-acetylhexosaminidases 183:207 arg1 characterization 153:168 characterization 153:168 characterization 153:168 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 2 theme β-N-acetylhexosaminidases 183:207 arg1 discovery 139:147 discovery 139:147 discovery 139:147 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 5 3 theme hydrolase 849:857 arg1 family 859:864 glycosyl hydrolase family 840:864 glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively 840:1079 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 0 4 from structures 9:18 arg1 oligosaccharides 34:49 human milk oligosaccharides 23:49 human milk oligosaccharides 23:49 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 3 5 theme N-terminal 576:585 arg1 His6-tag 587:594 an N-terminal His6-tag 573:594 an N-terminal His6-tag 573:594 The β-N-acetylhexosaminidases were expressed in Escherichia coli with an N-terminal His6-tag and were purified by nickel affinity chromatography. 25843303 4 6 from % 756:756 arg1 level 782:786 the protein level 770:786 the protein level 770:786 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 4 7 theme sequence 653:660 arg1 similarities 662:673 The sequence similarities 649:673 The sequence similarities of the enzymes with their respective closest homologues 649:729 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 4 7 theme sequence 653:660 arg1 % 738:738 59 % 735:738 59 % for HEX1 735:747 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 6 8 theme two 1181:1183 arg1 polymerization 1163:1176 polymerization 1163:1176 polymerization of two, three, and four 1163:1200 The enzymes are able to hydrolyze N-acetylchitooligosaccharides with a degree of polymerization of two, three, and four. 25843303 7 9 theme human 1331:1335 arg1 lacto-N-triose 1368:1381 the human milk oligosaccharide precursor lacto-N-triose II 1327:1384 the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate 1327:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 9 theme human 1331:1335 arg1 LNT2 1387:1390 LNT2 1387:1390 LNT2 1387:1390 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 2 10 link soil-derived 469:480 arg1 library 494:500 a soil-derived metagenomic library 467:500 a soil-derived metagenomic library 467:500 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 5 11 theme 6 1047:1047 arg1 optima 1031:1036 pH optima 1028:1036 pH optima of 8 and 6 for HEX1 and HEX2 1028:1065 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 8 12 attach attachment 1651:1660 arg2 GlcNAc 1665:1670 GlcNAc 1665:1670 GlcNAc 1665:1670 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 12 attach attachment 1651:1660 arg1 acceptor 1679:1686 the acceptor 1675:1686 the acceptor in all cases 1675:1699 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 3 13 theme nickel 617:622 arg1 chromatography 633:646 nickel affinity chromatography 617:646 nickel affinity chromatography 617:646 The β-N-acetylhexosaminidases were expressed in Escherichia coli with an N-terminal His6-tag and were purified by nickel affinity chromatography. 25843303 1 14 theme backbone 297:304 arg1 structures 306:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures 224:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor 224:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 14 theme backbone 297:304 arg1 β-N-acetylhexosaminidases 183:207 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 15 dep discovery 139:147 arg1 the 135:137 the 135:137 the 135:137 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 6 16 theme polymerization 1163:1176 arg1 degree 1153:1158 a degree 1151:1158 a degree of polymerization of two, three, and four 1151:1200 The enzymes are able to hydrolyze N-acetylchitooligosaccharides with a degree of polymerization of two, three, and four. 25843303 7 17 theme major 1207:1211 arg1 findings 1213:1220 The major findings 1203:1220 The major findings 1203:1220 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 8 18 dep monosaccharides 1585:1599 arg1 monosaccharides 1585:1599 the monosaccharides galactose and glucose 1581:1621 the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases 1581:1699 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 18 dep monosaccharides 1585:1599 arg1 glucose 1615:1621 glucose 1615:1621 glucose 1615:1621 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 18 dep monosaccharides 1585:1599 arg1 galactose 1601:1609 galactose 1601:1609 galactose 1601:1609 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 0 19 theme Backbone 0:7 arg1 structures 9:18 Backbone structures 0:18 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 0:112 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 5 20 dep family 859:864 arg1 exhibit 1020:1026 exhibit 1020:1026 exhibit 1020:1026 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 5 20 dep family 859:864 arg1 able 881:884 able 881:884 able 881:884 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 1 21 dep β-N-acetylhexosaminidases 183:207 arg1 β-N-acetylhexosaminidases 183:207 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 21 dep β-N-acetylhexosaminidases 183:207 arg1 HEX2 218:221 HEX2 218:221 HEX2 218:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 21 dep β-N-acetylhexosaminidases 183:207 arg1 HEX1 209:212 HEX1 209:212 HEX1 209:212 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 4 22 from % 738:738 arg1 level 782:786 the protein level 770:786 the protein level 770:786 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 2 23 theme library 494:500 arg1 screening 454:462 functional screening 443:462 functional screening of a soil-derived metagenomic library 443:500 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 1 24 with structures 306:315 arg1 yields 327:332 fair yields 322:332 fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor 322:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 25 theme capable 224:230 arg1 structures 306:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures 224:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor 224:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 25 theme capable 224:230 arg1 β-N-acetylhexosaminidases 183:207 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 0 26 theme human 23:27 arg1 oligosaccharides 34:49 human milk oligosaccharides 23:49 human milk oligosaccharides 23:49 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 4 27 with similarities 662:673 arg1 homologues 720:729 their respective closest homologues 695:729 their respective closest homologues 695:729 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 5 28 theme pH 1028:1029 arg1 optima 1031:1036 pH optima 1028:1036 pH optima of 8 and 6 for HEX1 and HEX2 1028:1065 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 3 29 theme affinity 624:631 arg1 chromatography 633:646 nickel affinity chromatography 617:646 nickel affinity chromatography 617:646 The β-N-acetylhexosaminidases were expressed in Escherichia coli with an N-terminal His6-tag and were purified by nickel affinity chromatography. 25843303 2 30 theme soil-derived 469:480 arg1 library 494:500 a soil-derived metagenomic library 467:500 a soil-derived metagenomic library 467:500 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 1 31 theme fair 322:325 arg1 yields 327:332 fair yields 322:332 fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor 322:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 7 32 with lacto-N-triose 1368:1381 arg1 yields 1398:1403 yields 1398:1403 yields of 2 and 8 % based on the donor substrate 1398:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 4 33 theme closest 712:718 arg1 homologues 720:729 their respective closest homologues 695:729 their respective closest homologues 695:729 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 7 34 theme trans-glycosylation 1256:1274 arg1 reactions 1276:1284 trans-glycosylation reactions 1256:1284 trans-glycosylation reactions with lactose 1256:1297 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 35 theme oligosaccharide 1342:1356 arg1 lacto-N-triose 1368:1381 the human milk oligosaccharide precursor lacto-N-triose II 1327:1384 the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate 1327:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 35 theme oligosaccharide 1342:1356 arg1 LNT2 1387:1390 LNT2 1387:1390 LNT2 1387:1390 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 36 theme precursor 1358:1366 arg1 lacto-N-triose 1368:1381 the human milk oligosaccharide precursor lacto-N-triose II 1327:1384 the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate 1327:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 36 theme precursor 1358:1366 arg1 LNT2 1387:1390 LNT2 1387:1390 LNT2 1387:1390 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 0 37 theme milk 29:32 arg1 oligosaccharides 34:49 human milk oligosaccharides 23:49 human milk oligosaccharides 23:49 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 7 38 theme donor 1431:1435 arg1 substrate 1437:1445 the donor substrate 1427:1445 the donor substrate 1427:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 1 39 dep capable 224:230 arg1 HMO 292:294 HMO 292:294 HMO 292:294 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 2 40 theme functional 443:452 arg1 screening 454:462 functional screening 443:462 functional screening of a soil-derived metagenomic library 443:500 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 1 41 theme chitin 340:345 arg1 oligomers 347:355 chitin oligomers 340:355 chitin oligomers 340:355 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 8 42 from acceptor 1679:1686 arg1 cases 1695:1699 all cases 1691:1699 all cases 1691:1699 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 7 43 theme % 1416:1416 arg1 yields 1398:1403 yields 1398:1403 yields of 2 and 8 % based on the donor substrate 1398:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 6 44 theme three 1186:1190 arg1 polymerization 1163:1176 polymerization 1163:1176 polymerization of two, three, and four 1163:1200 The enzymes are able to hydrolyze N-acetylchitooligosaccharides with a degree of polymerization of two, three, and four. 25843303 7 45 theme milk 1337:1340 arg1 lacto-N-triose 1368:1381 the human milk oligosaccharide precursor lacto-N-triose II 1327:1384 the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate 1327:1445 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 7 45 theme milk 1337:1340 arg1 LNT2 1387:1390 LNT2 1387:1390 LNT2 1387:1390 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 2 46 theme metagenomic 482:492 arg1 library 494:500 a soil-derived metagenomic library 467:500 a soil-derived metagenomic library 467:500 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 7 47 with reactions 1276:1284 arg1 lactose 1291:1297 lactose 1291:1297 lactose 1291:1297 The major findings were, that HEX1 and HEX2 catalyze trans-glycosylation reactions with lactose as acceptor, giving rise to the human milk oligosaccharide precursor lacto-N-triose II (LNT2) with yields of 2 and 8 % based on the donor substrate. 25843303 6 48 theme four 1197:1200 arg1 polymerization 1163:1176 polymerization 1163:1176 polymerization of two, three, and four 1163:1200 The enzymes are able to hydrolyze N-acetylchitooligosaccharides with a degree of polymerization of two, three, and four. 25843303 8 49 theme GlcNAc 1665:1670 arg1 attachment 1651:1660 the successful attachment 1636:1660 the successful attachment of GlcNAc to the acceptor in all cases 1636:1699 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 50 theme disaccharide 1509:1520 arg1 acceptors 1522:1530 the disaccharide acceptors β-lactose, sucrose, and maltose 1505:1562 the disaccharide acceptors β-lactose, sucrose, and maltose 1505:1562 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 50 theme disaccharide 1509:1520 arg1 sucrose 1543:1549 sucrose 1543:1549 sucrose 1543:1549 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 50 theme disaccharide 1509:1520 arg1 maltose 1556:1562 maltose 1556:1562 maltose 1556:1562 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 8 50 theme disaccharide 1509:1520 arg1 β-lactose 1532:1540 β-lactose 1532:1540 β-lactose 1532:1540 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 5 51 theme 8 1041:1041 arg1 optima 1031:1036 pH optima 1028:1036 pH optima of 8 and 6 for HEX1 and HEX2 1028:1065 Both β-N-acetylhexosaminidases are classified into glycosyl hydrolase family 20 (GH 20) are able to hydrolyze para-nitrophenyl-β-N-acetylglucosamine (pNP-GlcNAc) as well as para-nitrophenyl-β-N-acetylgalactosamine (pNP-GalNAc) and exhibit pH optima of 8 and 6 for HEX1 and HEX2, respectively. 25843303 8 52 theme successful 1640:1649 arg1 attachment 1651:1660 the successful attachment 1636:1660 the successful attachment of GlcNAc to the acceptor in all cases 1636:1699 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 4 53 theme protein 774:780 arg1 level 782:786 the protein level 770:786 the protein level 770:786 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 1 54 theme human 263:267 arg1 oligosaccharides 274:289 human milk oligosaccharides 263:289 human milk oligosaccharides 263:289 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 0 55 theme metagenomic 75:85 arg1 β-N-acetylhexosaminidases 87:111 metagenomic β-N-acetylhexosaminidases 75:111 metagenomic β-N-acetylhexosaminidases 75:111 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 0 56 dep structures 9:18 arg1 trans-glycosylation 52:70 trans-glycosylation 52:70 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 0:112 Backbone structures in human milk oligosaccharides: trans-glycosylation by metagenomic β-N-acetylhexosaminidases. 25843303 4 57 theme respective 701:710 arg1 homologues 720:729 their respective closest homologues 695:729 their respective closest homologues 695:729 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 2 58 theme enzyme-encoding 402:416 arg1 genes 418:422 The enzyme-encoding genes 398:422 The enzyme-encoding genes 398:422 The enzyme-encoding genes were identified by functional screening of a soil-derived metagenomic library. 25843303 1 59 theme milk 269:272 arg1 oligosaccharides 274:289 human milk oligosaccharides 263:289 human milk oligosaccharides 263:289 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 60 theme β-N-acetylglucosamine 360:380 arg1 donor 391:395 β-N-acetylglucosamine (GlcNAc) donor 360:395 β-N-acetylglucosamine (GlcNAc) donor 360:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 8 61 theme trans-glycosylation 1458:1476 arg1 reactions 1478:1486 trans-glycosylation reactions 1458:1486 trans-glycosylation reactions 1458:1486 In total, trans-glycosylation reactions were tested with the disaccharide acceptors β-lactose, sucrose, and maltose, as well as with the monosaccharides galactose and glucose resulting in the successful attachment of GlcNAc to the acceptor in all cases. 25843303 4 62 theme enzymes 682:688 arg1 similarities 662:673 The sequence similarities 649:673 The sequence similarities of the enzymes with their respective closest homologues 649:729 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 4 62 theme enzymes 682:688 arg1 % 738:738 59 % 735:738 59 % for HEX1 735:747 The sequence similarities of the enzymes with their respective closest homologues are 59 % for HEX1 and 51 % for HEX2 on the protein level. 25843303 1 63 theme novel 177:181 arg1 structures 306:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures 224:315 capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor 224:395 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 63 theme novel 177:181 arg1 β-N-acetylhexosaminidases 183:207 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 two novel β-N-acetylhexosaminidases HEX1 and HEX2 173:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 63 theme novel 177:181 arg1 HEX2 218:221 HEX2 218:221 HEX2 218:221 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 63 theme novel 177:181 arg1 HEX1 209:212 HEX1 209:212 HEX1 209:212 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 25843303 1 64 theme oligosaccharides 274:289 arg1 synthesis 250:258 the synthesis 246:258 the synthesis of human milk oligosaccharides 246:289 This paper describes the discovery and characterization of two novel β-N-acetylhexosaminidases HEX1 and HEX2, capable of catalyzing the synthesis of human milk oligosaccharides (HMO) backbone structures with fair yields using chitin oligomers as β-N-acetylglucosamine (GlcNAc) donor. 26211613 0 0 from Role 15:18 arg1 Interactions 60:71 Receptor Interactions 51:71 Receptor Interactions 51:71 The Structural Role of Antibody N-Glycosylation in Receptor Interactions. 26211613 3 1 theme E 458:458 arg1 loop 460:463 the C'E loop 452:463 the C'E loop 452:463 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 5 2 theme primary 704:710 arg1 role 712:715 the primary role 700:715 the primary role of the IgG1 Fc N-glycan 700:739 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 4 3 with correlation 636:646 arg1 affinity 662:669 FcγRIIIa affinity 653:669 FcγRIIIa affinity 653:669 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 3 4 theme loop 460:463 arg1 conformation 436:447 the conformation 432:447 the conformation of the C'E loop, which contains N297, 432:485 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 4 5 dep loop 576:579 arg1 measured 584:591 measured 584:591 measured with a range of Fc variants 584:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 1 6 dep G 130:130 arg1 Fc 138:139 Fc 138:139 immunoglobulin G (IgG) Fc 115:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 6 7 theme other 1066:1070 arg1 IgM 1118:1120 IgM 1118:1120 IgM 1118:1120 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 6 7 theme other 1066:1070 arg1 IgD 1104:1106 IgD 1104:1106 IgD 1104:1106 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 6 7 theme other 1066:1070 arg1 IgG2-IgG4 1093:1101 IgG2-IgG4 1093:1101 IgG2-IgG4 1093:1101 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 6 7 theme other 1066:1070 arg1 antibodies 1072:1081 other antibodies 1066:1081 other antibodies including IgG2-IgG4, IgD, IgE, and IgM 1066:1120 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 6 7 theme other 1066:1070 arg1 IgE 1109:1111 IgE 1109:1111 IgE 1109:1111 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 5 8 theme optimal 895:901 arg1 affinity 911:918 optimal binding affinity 895:918 optimal binding affinity 895:918 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 4 9 theme FcγRIIIa 653:660 arg1 affinity 662:669 FcγRIIIa affinity 653:669 FcγRIIIa affinity 653:669 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 1 10 gly glycosylation 98:110 arg1 G 130:130 immunoglobulin G 115:130 immunoglobulin G (IgG) Fc 115:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 1 10 gly glycosylation 98:110 arg1 IgG 133:135 IgG 133:135 IgG 133:135 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 5 11 theme Fc 729:730 arg1 N-glycan 732:739 the IgG1 Fc N-glycan 720:739 the IgG1 Fc N-glycan 720:739 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 5 12 theme binding 903:909 arg1 affinity 911:918 optimal binding affinity 895:918 optimal binding affinity 895:918 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 6 13 theme IgG1 973:976 arg1 N-glycan 981:988 the IgG1 Fc N-glycan 969:988 the IgG1 Fc N-glycan 969:988 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 5 14 theme amino 831:835 arg1 residues 842:849 amino acid residues 831:849 amino acid residues 831:849 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 5 15 theme N-glycan 732:739 arg1 role 712:715 the primary role 700:715 the primary role of the IgG1 Fc N-glycan 700:739 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 0 16 theme Structural 4:13 arg1 Role 15:18 The Structural Role 0:18 The Structural Role of Antibody N-Glycosylation in Receptor Interactions 0:71 The Structural Role of Antibody N-Glycosylation in Receptor Interactions. 26211613 6 17 theme protein 1002:1008 arg1 conformation 1010:1021 protein conformation 1002:1021 protein conformation 1002:1021 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 5 18 theme intramolecular 778:791 arg1 interactions 793:804 intramolecular interactions 778:804 intramolecular interactions between carbohydrate and amino acid residues 778:849 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 2 19 theme domain 403:408 arg1 orientation 410:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 20 from Fc 329:330 arg1 indistinguishable 292:308 indistinguishable 292:308 indistinguishable 292:308 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 20 from Fc 329:330 arg1 structure 259:267 the quaternary structure 244:267 the quaternary structure of glycosylated Fc 244:286 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 21 theme quaternary 248:257 arg1 indistinguishable 292:308 indistinguishable 292:308 indistinguishable 292:308 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 21 theme quaternary 248:257 arg1 structure 259:267 the quaternary structure 244:267 the quaternary structure of glycosylated Fc 244:286 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 6 22 theme N-glycan 981:988 arg1 capacity 957:964 the capacity 953:964 the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity 953:1047 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 2 23 theme Cγ2/Cγ3 395:401 arg1 orientation 410:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 24 gly aglycosylated 315:327 arg1 Fc 329:330 aglycosylated Fc 315:330 aglycosylated Fc 315:330 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 1 25 theme 297-linked 87:96 arg1 glycosylation 98:110 Asparagine(N)297-linked glycosylation 74:110 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc 74:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 2 26 theme relative 383:390 arg1 orientation 410:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 27 theme Fc 392:393 arg1 orientation 410:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 relative Fc Cγ2/Cγ3 domain orientation 383:420 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 3 28 theme C' 456:457 arg1 loop 460:463 the C'E loop 452:463 the C'E loop 452:463 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 4 29 theme E 574:574 arg1 loop 576:579 the C'E loop 568:579 the C'E loop as measured with a range of Fc variants 568:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 0 30 theme N-Glycosylation 32:46 arg1 Role 15:18 The Structural Role 0:18 The Structural Role of Antibody N-Glycosylation in Receptor Interactions 0:71 The Structural Role of Antibody N-Glycosylation in Receptor Interactions. 26211613 1 31 theme immunoglobulin 115:128 arg1 G 130:130 immunoglobulin G 115:130 immunoglobulin G (IgG) Fc 115:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 1 31 theme immunoglobulin 115:128 arg1 IgG 133:135 IgG 133:135 IgG 133:135 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 5 32 theme C' 761:762 arg1 loop 765:768 the C'E loop 757:768 the C'E loop 757:768 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 0 33 theme Antibody 23:30 arg1 N-Glycosylation 32:46 Antibody N-Glycosylation 23:46 Antibody N-Glycosylation 23:46 The Structural Role of Antibody N-Glycosylation in Receptor Interactions. 26211613 3 34 gly aglycosylated 522:534 arg1 Fc variant 536:545 the aglycosylated Fc variant 518:545 the aglycosylated Fc variant 518:545 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 0 35 theme Receptor 51:58 arg1 Interactions 60:71 Receptor Interactions 51:71 Receptor Interactions 51:71 The Structural Role of Antibody N-Glycosylation in Receptor Interactions. 26211613 3 36 contain contains 472:479 arg1 loop 460:463 the C'E loop 452:463 the C'E loop 452:463 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 3 36 contain contains 472:479 arg2 N297 481:484 N297 481:484 N297 481:484 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 3 37 theme aglycosylated 522:534 arg1 Fc variant 536:545 the aglycosylated Fc variant 518:545 the aglycosylated Fc variant 518:545 However, the conformation of the C'E loop, which contains N297, was significantly perturbed in the aglycosylated Fc variant. 26211613 6 38 theme binding 1032:1038 arg1 affinity 1040:1047 binding affinity 1032:1047 binding affinity 1032:1047 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 5 39 theme acid 837:840 arg1 residues 842:849 amino acid residues 831:849 amino acid residues 831:849 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 5 40 theme IgG1 724:727 arg1 Fc 729:730 IgG1 Fc 724:730 the IgG1 Fc N-glycan 720:739 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 2 41 theme Fc 285:286 arg1 indistinguishable 292:308 indistinguishable 292:308 indistinguishable 292:308 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 2 41 theme Fc 285:286 arg1 structure 259:267 the quaternary structure 244:267 the quaternary structure of glycosylated Fc 244:286 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 1 42 theme G 130:130 arg1 glycosylation 98:110 Asparagine(N)297-linked glycosylation 74:110 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc 74:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 5 43 theme FcγRIIIa 872:879 arg1 interface 881:889 the FcγRIIIa interface 868:889 the FcγRIIIa interface for optimal binding affinity 868:918 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 2 44 gly glycosylated 272:283 arg1 Fc 285:286 glycosylated Fc 272:286 glycosylated Fc 272:286 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 6 45 theme Fc 978:979 arg1 N-glycan 981:988 the IgG1 Fc N-glycan 969:988 the IgG1 Fc N-glycan 969:988 The features that contribute to the capacity of the IgG1 Fc N-glycan to restrict protein conformation and tune binding affinity are conserved in other antibodies including IgG2-IgG4, IgD, IgE, and IgM. 26211613 4 46 theme Fc 609:610 arg1 variants 612:619 Fc variants 609:619 Fc variants 609:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 2 47 theme glycosylated 272:283 arg1 Fc 285:286 glycosylated Fc 272:286 glycosylated Fc 272:286 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 1 48 link 297-linked 87:96 arg1 glycosylation 98:110 Asparagine(N)297-linked glycosylation 74:110 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc 74:139 Asparagine(N)297-linked glycosylation of immunoglobulin G (IgG) Fc is required for binding to FcγRIIa, IIb, and IIIa, although it is unclear how it contributes. 26211613 5 49 theme E 763:763 arg1 loop 765:768 the C'E loop 757:768 the C'E loop 757:768 These results indicate that the primary role of the IgG1 Fc N-glycan is to stabilize the C'E loop through intramolecular interactions between carbohydrate and amino acid residues, and preorganize the FcγRIIIa interface for optimal binding affinity. 26211613 4 50 theme loop 576:579 arg1 conformation 552:563 The conformation 548:563 The conformation of the C'E loop as measured with a range of Fc variants 548:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 4 51 theme variants 612:619 arg1 range 600:604 a range 598:604 a range of Fc variants 598:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 4 52 theme C' 572:573 arg1 loop 576:579 the C'E loop 568:579 the C'E loop as measured with a range of Fc variants 568:619 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26211613 2 53 theme aglycosylated 315:327 arg1 Fc 329:330 aglycosylated Fc 315:330 aglycosylated Fc 315:330 We found the quaternary structure of glycosylated Fc was indistinguishable from aglycosylated Fc, indicating that N-glycosylation does not maintain relative Fc Cγ2/Cγ3 domain orientation. 26211613 4 54 theme strong 629:634 arg1 correlation 636:646 a strong correlation 627:646 a strong correlation with FcγRIIIa affinity 627:669 The conformation of the C'E loop as measured with a range of Fc variants shows a strong correlation with FcγRIIIa affinity. 26478188 7 0 theme CMP-Ang1 1198:1205 arg1 effects 1179:1185 these effects 1173:1185 these effects of dimeric CMP-Ang1 1173:1205 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 7 1 gly N-glycosylation 1234:1248 arg1 domain 1273:1278 its fibrinogen-like domain 1253:1278 its fibrinogen-like domain 1253:1278 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 3 2 theme native 654:659 arg1 Ang1 661:664 native Ang1 654:664 native Ang1 654:664 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 8 3 theme dimeric 1323:1329 arg1 CMP-Ang1 1331:1338 dimeric CMP-Ang1 1323:1338 dimeric CMP-Ang1 1323:1338 Taken together, our results indicate that dimeric CMP-Ang1 is capable of activating Tie2 and stimulating angiogenesis in N-glycan dependent manner. 26478188 0 4 theme Vascular 99:106 arg1 Stabilization 108:120 Vascular Stabilization 99:120 Vascular Stabilization 99:120 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 3 5 theme Ang1 661:664 arg1 N-terminal 640:649 N-terminal 640:649 N-terminal 640:649 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 6 from mutations 736:744 arg1 residues 762:769 its cysteine residues 749:769 its cysteine residues 749:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 7 7 from N-glycosylation 1234:1248 arg1 domain 1273:1278 its fibrinogen-like domain 1253:1278 its fibrinogen-like domain 1253:1278 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 7 8 theme dimeric 1190:1196 arg1 CMP-Ang1 1198:1205 dimeric CMP-Ang1 1190:1205 dimeric CMP-Ang1 1190:1205 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 1 9 theme unique 346:351 arg1 forms 328:332 high oligomeric forms 312:332 high oligomeric forms 312:332 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 9 theme unique 346:351 arg1 process 367:373 a unique and essential process 344:373 a unique and essential process in this ligand-receptor interaction 344:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 7 10 theme specified 1224:1232 arg1 N-glycosylation 1234:1248 specified N-glycosylation 1224:1248 specified N-glycosylation in its fibrinogen-like domain 1224:1278 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 3 11 theme cartilage 697:705 arg1 CMP 723:725 CMP 723:725 CMP 723:725 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 11 theme cartilage 697:705 arg1 protein 714:720 cartilage matrix protein 697:720 cartilage matrix protein (CMP) bearing mutations in its cysteine residues 697:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 6 12 theme such 1042:1045 arg1 effects 1071:1077 such Tie2-activation-induced effects 1042:1077 such Tie2-activation-induced effects 1042:1077 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 6 13 theme effects 1071:1077 arg1 potencies 1029:1037 the potencies 1025:1037 the potencies of such Tie2-activation-induced effects 1025:1077 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 6 13 theme effects 1071:1077 arg1 comparable 1084:1093 comparable 1084:1093 comparable 1084:1093 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 1 14 theme essential 357:365 arg1 forms 328:332 high oligomeric forms 312:332 high oligomeric forms 312:332 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 14 theme essential 357:365 arg1 process 367:373 a unique and essential process 344:373 a unique and essential process in this ligand-receptor interaction 344:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 4 15 dep migration 820:828 arg1 the 816:818 the 816:818 the 816:818 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 6 16 theme Tie2-activation-induced 1047:1069 arg1 effects 1071:1077 such Tie2-activation-induced effects 1042:1077 such Tie2-activation-induced effects 1042:1077 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 0 17 theme Designed 2:9 arg1 Variant 26:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 0 17 theme Designed 2:9 arg1 CMP-Ang1 43:50 Dimeric CMP-Ang1 35:50 Dimeric CMP-Ang1 35:50 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 1 18 theme potential 177:185 arg1 Angiopoietin-1 152:165 Angiopoietin-1 152:165 Angiopoietin-1 (Ang1) 152:172 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 18 theme potential 177:185 arg1 factor 194:199 a potential growth factor 175:199 a potential growth factor for therapeutic angiogenesis and vascular stabilization 175:255 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 19 from process 367:373 arg1 interaction 399:409 this ligand-receptor interaction 378:409 this ligand-receptor interaction 378:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 20 theme growth 187:192 arg1 Angiopoietin-1 152:165 Angiopoietin-1 152:165 Angiopoietin-1 (Ang1) 152:172 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 20 theme growth 187:192 arg1 factor 194:199 a potential growth factor 175:199 a potential growth factor for therapeutic angiogenesis and vascular stabilization 175:255 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 2 21 theme native 439:444 arg1 Ang1 446:449 highly oligomeric native Ang1 421:449 highly oligomeric native Ang1 421:449 However, highly oligomeric native Ang1 and Ang1 variants are difficult to produce, purify, and store in a stable and active form. 26478188 0 22 theme Dependent 134:142 arg1 Manner 144:149 N-glycan Dependent Manner 125:149 N-glycan Dependent Manner 125:149 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 5 23 theme dimeric 915:921 arg1 CMP-Ang1 923:930 dimeric CMP-Ang1 915:930 dimeric CMP-Ang1 915:930 Furthermore, dimeric CMP-Ang1 induced angiogenesis and suppressed vascular leakage in vivo. 26478188 2 24 theme oligomeric 428:437 arg1 Ang1 446:449 highly oligomeric native Ang1 421:449 highly oligomeric native Ang1 421:449 However, highly oligomeric native Ang1 and Ang1 variants are difficult to produce, purify, and store in a stable and active form. 26478188 3 25 theme matrix 707:712 arg1 CMP 723:725 CMP 723:725 CMP 723:725 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 25 theme matrix 707:712 arg1 protein 714:720 cartilage matrix protein 697:720 cartilage matrix protein (CMP) bearing mutations in its cysteine residues 697:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 0 26 from Angiogenesis 82:93 arg1 Manner 144:149 N-glycan Dependent Manner 125:149 N-glycan Dependent Manner 125:149 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 0 27 theme Dimeric 35:41 arg1 Variant 26:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 0 27 theme Dimeric 35:41 arg1 CMP-Ang1 43:50 Dimeric CMP-Ang1 35:50 Dimeric CMP-Ang1 35:50 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 3 28 theme protein 714:720 arg1 domain 687:692 the coiled-coil domain 671:692 the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues 671:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 28 theme protein 714:720 arg1 protein 714:720 cartilage matrix protein 697:720 cartilage matrix protein (CMP) bearing mutations in its cysteine residues 697:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 2 29 theme stable 518:523 arg1 form 536:539 a stable and active form 516:539 a stable and active form 516:539 However, highly oligomeric native Ang1 and Ang1 variants are difficult to produce, purify, and store in a stable and active form. 26478188 4 30 theme dimeric 777:783 arg1 CMP-Ang1 785:792 This dimeric CMP-Ang1 772:792 This dimeric CMP-Ang1 772:792 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 1 31 theme therapeutic 205:215 arg1 angiogenesis 217:228 therapeutic angiogenesis 205:228 therapeutic angiogenesis 205:228 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 32 theme ligand-receptor 383:397 arg1 interaction 399:409 this ligand-receptor interaction 378:409 this ligand-receptor interaction 378:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 33 theme high 312:315 arg1 process 367:373 a unique and essential process 344:373 a unique and essential process in this ligand-receptor interaction 344:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 33 theme high 312:315 arg1 forms 328:332 high oligomeric forms 312:332 high oligomeric forms 312:332 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 4 34 theme tube 845:848 arg1 formation 850:858 tube formation 845:858 tube formation 845:858 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 3 35 theme dimeric 606:612 arg1 CMP-Ang1 614:621 a simple and active dimeric CMP-Ang1 586:621 a simple and active dimeric CMP-Ang1 586:621 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 36 theme coiled-coil 675:685 arg1 domain 687:692 the coiled-coil domain 671:692 the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues 671:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 3 36 theme coiled-coil 675:685 arg1 protein 714:720 cartilage matrix protein 697:720 cartilage matrix protein (CMP) bearing mutations in its cysteine residues 697:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 6 37 theme engineered 1120:1129 arg1 COMP-Ang1 1140:1148 COMP-Ang1 1140:1148 COMP-Ang1 1140:1148 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 6 37 theme engineered 1120:1129 arg1 protein 1131:1137 a previously engineered protein 1107:1137 a previously engineered protein 1107:1137 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 2 38 theme Ang1 455:458 arg1 variants 460:467 Ang1 variants 455:467 Ang1 variants 455:467 However, highly oligomeric native Ang1 and Ang1 variants are difficult to produce, purify, and store in a stable and active form. 26478188 1 39 theme oligomeric 317:326 arg1 process 367:373 a unique and essential process 344:373 a unique and essential process in this ligand-receptor interaction 344:409 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 1 39 theme oligomeric 317:326 arg1 forms 328:332 high oligomeric forms 312:332 high oligomeric forms 312:332 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 2 40 theme active 529:534 arg1 form 536:539 a stable and active form 516:539 a stable and active form 516:539 However, highly oligomeric native Ang1 and Ang1 variants are difficult to produce, purify, and store in a stable and active form. 26478188 8 41 theme dependent 1411:1419 arg1 manner 1421:1426 N-glycan dependent manner 1402:1426 N-glycan dependent manner 1402:1426 Taken together, our results indicate that dimeric CMP-Ang1 is capable of activating Tie2 and stimulating angiogenesis in N-glycan dependent manner. 26478188 3 42 theme active 599:604 arg1 CMP-Ang1 614:621 a simple and active dimeric CMP-Ang1 586:621 a simple and active dimeric CMP-Ang1 586:621 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 4 43 theme cells 875:879 arg1 migration 820:828 migration 820:828 migration 820:828 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 4 43 theme cells 875:879 arg1 formation 850:858 tube formation 845:858 tube formation 845:858 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 4 43 theme cells 875:879 arg1 survival 831:838 survival 831:838 survival 831:838 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 0 44 from Stabilization 108:120 arg1 Manner 144:149 N-glycan Dependent Manner 125:149 N-glycan Dependent Manner 125:149 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 3 45 theme simple 588:593 arg1 CMP-Ang1 614:621 a simple and active dimeric CMP-Ang1 586:621 a simple and active dimeric CMP-Ang1 586:621 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 26478188 7 46 theme fibrinogen-like 1257:1271 arg1 domain 1273:1278 its fibrinogen-like domain 1253:1278 its fibrinogen-like domain 1253:1278 We also revealed that these effects of dimeric CMP-Ang1 were affected by specified N-glycosylation in its fibrinogen-like domain. 26478188 6 47 theme dimeric 1006:1012 arg1 structure 1014:1022 its dimeric structure 1002:1022 its dimeric structure 1002:1022 Despite its dimeric structure, the potencies of such Tie2-activation-induced effects were comparable to those of a previously engineered protein, COMP-Ang1. 26478188 4 48 theme endothelial 863:873 arg1 cells 875:879 endothelial cells 863:879 endothelial cells 863:879 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 1 49 theme vascular 234:241 arg1 stabilization 243:255 vascular stabilization 234:255 vascular stabilization 234:255 Angiopoietin-1 (Ang1), a potential growth factor for therapeutic angiogenesis and vascular stabilization, is known to specifically cluster and activate Tie2 in high oligomeric forms, which is a unique and essential process in this ligand-receptor interaction. 26478188 4 50 theme Tie2 885:888 arg1 activation 890:899 Tie2 activation 885:899 Tie2 activation 885:899 This dimeric CMP-Ang1 effectively increased the migration, survival, and tube formation of endothelial cells via Tie2 activation. 26478188 5 51 theme vascular 968:975 arg1 leakage 977:983 vascular leakage 968:983 vascular leakage 968:983 Furthermore, dimeric CMP-Ang1 induced angiogenesis and suppressed vascular leakage in vivo. 26478188 0 52 theme Angiopoietin-1 11:24 arg1 Variant 26:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant 0:32 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 0 52 theme Angiopoietin-1 11:24 arg1 CMP-Ang1 43:50 Dimeric CMP-Ang1 35:50 Dimeric CMP-Ang1 35:50 A Designed Angiopoietin-1 Variant, Dimeric CMP-Ang1 Activates Tie2 and Stimulates Angiogenesis and Vascular Stabilization in N-glycan Dependent Manner. 26478188 8 53 from angiogenesis 1386:1397 arg1 manner 1421:1426 N-glycan dependent manner 1402:1426 N-glycan dependent manner 1402:1426 Taken together, our results indicate that dimeric CMP-Ang1 is capable of activating Tie2 and stimulating angiogenesis in N-glycan dependent manner. 26478188 3 54 theme cysteine 753:760 arg1 residues 762:769 its cysteine residues 749:769 its cysteine residues 749:769 To overcome these limitations, we developed a simple and active dimeric CMP-Ang1 by replacing the N-terminal of native Ang1 with the coiled-coil domain of cartilage matrix protein (CMP) bearing mutations in its cysteine residues. 28286305 0 0 theme Yip 82:84 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 0 theme Yip 82:84 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 0 theme Yip 82:84 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 0 theme Yip 82:84 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 3 1 theme TGN 663:665 arg1 marker 667:672 a TGN marker 661:672 a TGN marker in earlier time point 661:694 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 0 2 theme glycan 156:161 arg1 synthesis 163:171 glycan synthesis 156:171 glycan synthesis 156:171 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 11 3 theme glycan 1862:1867 arg1 synthesis 1869:1877 normal glycan synthesis 1855:1877 normal glycan synthesis 1855:1877 Thus, we confirmed that YIPF1, YIPF2, and YIPF6 play a significant role in supporting normal glycan synthesis. 28286305 6 4 dep expression 1178:1187 arg1 the 1174:1176 the 1174:1176 the 1174:1176 Knockdown experiments showed that YIPF1 and YIPF2, by contrast, are not necessary for the expression and localization of YIPF6. 28286305 0 5 theme network-localized 64:80 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 5 theme network-localized 64:80 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 5 theme network-localized 64:80 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 5 theme network-localized 64:80 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 10 6 theme HT-29 1756:1760 arg1 cells 1762:1766 HT-29 cells 1756:1766 HT-29 cells 1756:1766 Knockdown of YIPF1 and YIPF2, but not that of YIPF6, also reduced intracellular glycans in HT-29 cells. 28286305 1 7 theme Yif1p 376:380 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 7 theme Yif1p 376:380 arg1 homologs 364:371 homologs 364:371 homologs of Yif1p 364:380 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 8 8 theme apparatus 1402:1410 arg1 reassembly 1378:1387 reassembly 1378:1387 reassembly of the Golgi apparatus after the removal of BFA 1378:1435 However, reassembly of the Golgi apparatus after the removal of BFA was markedly delayed by the knockdown of YIPF1 and YIPF2, but not by that of YIPF6. 28286305 6 9 theme YIPF6 1209:1213 arg1 localization 1193:1204 localization 1193:1204 localization 1193:1204 Knockdown experiments showed that YIPF1 and YIPF2, by contrast, are not necessary for the expression and localization of YIPF6. 28286305 6 9 theme YIPF6 1209:1213 arg1 expression 1178:1187 expression 1178:1187 expression 1178:1187 Knockdown experiments showed that YIPF1 and YIPF2, by contrast, are not necessary for the expression and localization of YIPF6. 28286305 0 10 theme family 93:98 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 10 theme family 93:98 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 10 theme family 93:98 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 10 theme family 93:98 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 7 11 theme BFA 1279:1281 arg1 treatment 1283:1291 BFA treatment 1279:1291 BFA treatment 1279:1291 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 3 12 theme TGN 814:816 arg1 markers 818:824 the TGN markers 810:824 the TGN markers 810:824 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 3 13 theme punctate 742:749 arg1 structures 751:760 cytoplasmic punctate structures 730:760 cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers 730:824 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 0 14 theme domain 86:91 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 14 theme domain 86:91 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 14 theme domain 86:91 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 14 theme domain 86:91 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 7 15 theme YIPF6 1362:1366 arg1 knockdown 1332:1340 the knockdown 1328:1340 the knockdown of YIPF1, YIPF2, or YIPF6 1328:1366 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 7 16 theme YIPF2 1352:1356 arg1 knockdown 1332:1340 the knockdown 1328:1340 the knockdown of YIPF1, YIPF2, or YIPF6 1328:1366 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 9 17 theme apparatus 1654:1662 arg1 reassembly 1630:1639 the reassembly 1626:1639 the reassembly of the Golgi apparatus 1626:1662 These results strongly suggest that free YIPF6 after disassociating with YIPF1 and YIPF2 interferes with the reassembly of the Golgi apparatus. 28286305 1 18 theme Yip1p 326:330 arg1 YIPF2 347:351 YIPF2 347:351 YIPF2 347:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 18 theme Yip1p 326:330 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 18 theme Yip1p 326:330 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 18 theme Yip1p 326:330 arg1 homolog 315:321 a homolog 313:321 a homolog of Yip1p 313:330 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 18 theme Yip1p 326:330 arg1 YIPF1 337:341 YIPF1 337:341 YIPF1 337:341 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 3 19 from markers 818:824 arg1 distinct 772:779 distinct 772:779 distinct 772:779 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 5 20 with complexes 984:992 arg1 YIPF1 999:1003 YIPF1 999:1003 YIPF1 999:1003 These results suggest that YIPF6 forms complexes with YIPF1 and YIPF2 for their stable expression and localization within the Golgi apparatus. 28286305 5 20 with complexes 984:992 arg1 YIPF2 1009:1013 YIPF2 1009:1013 YIPF2 1009:1013 These results suggest that YIPF6 forms complexes with YIPF1 and YIPF2 for their stable expression and localization within the Golgi apparatus. 28286305 7 21 theme apparatus 1243:1251 arg1 structure 1220:1228 The structure 1216:1228 The structure of the Golgi apparatus 1216:1251 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 7 21 theme apparatus 1243:1251 arg1 disassembly 1261:1271 its disassembly 1257:1271 its disassembly after BFA treatment 1257:1291 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 5 22 theme stable 1025:1030 arg1 expression 1032:1041 their stable expression 1019:1041 their stable expression 1019:1041 These results suggest that YIPF6 forms complexes with YIPF1 and YIPF2 for their stable expression and localization within the Golgi apparatus. 28286305 3 23 from marker 667:672 arg1 point 690:694 earlier time point 677:694 earlier time point 677:694 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 7 24 theme YIPF1 1345:1349 arg1 knockdown 1332:1340 the knockdown 1328:1340 the knockdown of YIPF1, YIPF2, or YIPF6 1328:1366 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 3 25 theme time 685:688 arg1 point 690:694 earlier time point 677:694 earlier time point 677:694 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 4 26 theme YIPF6 907:911 arg1 knockdown 894:902 knockdown 894:902 knockdown of YIPF6 894:911 YIPF6 formed a stable complex separately with YIPF1 and YIPF2, and knockdown of YIPF6 reduced YIPF1 and YIPF2 levels. 28286305 8 27 theme YIPF1 1478:1482 arg1 knockdown 1465:1473 the knockdown 1461:1473 the knockdown of YIPF1 and YIPF2 1461:1492 However, reassembly of the Golgi apparatus after the removal of BFA was markedly delayed by the knockdown of YIPF1 and YIPF2, but not by that of YIPF6. 28286305 6 28 theme Knockdown 1088:1096 arg1 experiments 1098:1108 Knockdown experiments 1088:1108 Knockdown experiments 1088:1108 Knockdown experiments showed that YIPF1 and YIPF2, by contrast, are not necessary for the expression and localization of YIPF6. 28286305 2 29 theme Immunofluorescence 383:400 arg1 staining 402:409 Immunofluorescence staining 383:409 Immunofluorescence staining 383:409 Immunofluorescence staining revealed that YIPF1, YIPF2, and YIPF6 mainly localize in the medial-/trans-Golgi and also partially in the trans-Golgi network (TGN). 28286305 1 30 theme uncharacterized 235:249 arg1 YIPF2 347:351 YIPF2 347:351 YIPF2 347:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 30 theme uncharacterized 235:249 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 30 theme uncharacterized 235:249 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 30 theme uncharacterized 235:249 arg1 homologs 364:371 homologs 364:371 homologs of Yif1p 364:380 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 30 theme uncharacterized 235:249 arg1 YIPF1 337:341 YIPF1 337:341 YIPF1 337:341 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 30 theme uncharacterized 235:249 arg1 homolog 315:321 a homolog 313:321 a homolog of Yip1p 313:330 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 9 31 theme free 1557:1560 arg1 YIPF6 1562:1566 free YIPF6 1557:1566 free YIPF6 after disassociating with YIPF1 and YIPF2 1557:1608 These results strongly suggest that free YIPF6 after disassociating with YIPF1 and YIPF2 interferes with the reassembly of the Golgi apparatus. 28286305 7 32 theme Golgi 1237:1241 arg1 apparatus 1243:1251 the Golgi apparatus 1233:1251 the Golgi apparatus 1233:1251 The structure of the Golgi apparatus and its disassembly after BFA treatment were not significantly affected by the knockdown of YIPF1, YIPF2, or YIPF6. 28286305 1 33 theme mammalian 251:259 arg1 YIPF2 347:351 YIPF2 347:351 YIPF2 347:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 33 theme mammalian 251:259 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 33 theme mammalian 251:259 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 33 theme mammalian 251:259 arg1 homologs 364:371 homologs 364:371 homologs of Yif1p 364:380 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 33 theme mammalian 251:259 arg1 YIPF1 337:341 YIPF1 337:341 YIPF1 337:341 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 33 theme mammalian 251:259 arg1 homolog 315:321 a homolog 313:321 a homolog of Yip1p 313:330 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 3 34 from medial-/trans-Golgi 786:804 arg1 distinct 772:779 distinct 772:779 distinct 772:779 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 8 35 theme Golgi 1396:1400 arg1 apparatus 1402:1410 the Golgi apparatus 1392:1410 the Golgi apparatus 1392:1410 However, reassembly of the Golgi apparatus after the removal of BFA was markedly delayed by the knockdown of YIPF1 and YIPF2, but not by that of YIPF6. 28286305 1 36 theme homologs 261:268 arg1 function 217:224 the function 213:224 the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p 213:380 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 11 37 theme normal 1855:1860 arg1 synthesis 1869:1877 normal glycan synthesis 1855:1877 normal glycan synthesis 1855:1877 Thus, we confirmed that YIPF1, YIPF2, and YIPF6 play a significant role in supporting normal glycan synthesis. 28286305 9 38 with YIPF6 1562:1566 arg1 YIPF2 1604:1608 YIPF2 1604:1608 YIPF2 1604:1608 These results strongly suggest that free YIPF6 after disassociating with YIPF1 and YIPF2 interferes with the reassembly of the Golgi apparatus. 28286305 9 38 with YIPF6 1562:1566 arg1 YIPF1 1594:1598 YIPF1 1594:1598 YIPF1 1594:1598 These results strongly suggest that free YIPF6 after disassociating with YIPF1 and YIPF2 interferes with the reassembly of the Golgi apparatus. 28286305 4 39 theme YIPF1 921:925 arg1 levels 937:942 YIPF1 and YIPF2 levels 921:942 YIPF1 and YIPF2 levels 921:942 YIPF6 formed a stable complex separately with YIPF1 and YIPF2, and knockdown of YIPF6 reduced YIPF1 and YIPF2 levels. 28286305 1 40 theme yeast 273:277 arg1 YIPF2 347:351 YIPF2 347:351 YIPF2 347:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 40 theme yeast 273:277 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 40 theme yeast 273:277 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 40 theme yeast 273:277 arg1 homologs 364:371 homologs 364:371 homologs of Yif1p 364:380 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 40 theme yeast 273:277 arg1 YIPF1 337:341 YIPF1 337:341 YIPF1 337:341 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 40 theme yeast 273:277 arg1 homolog 315:321 a homolog 313:321 a homolog of Yip1p 313:330 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 10 41 theme YIPF1 1678:1682 arg1 Knockdown 1665:1673 Knockdown 1665:1673 Knockdown of YIPF1 and YIPF2 1665:1692 Knockdown of YIPF1 and YIPF2, but not that of YIPF6, also reduced intracellular glycans in HT-29 cells. 28286305 0 42 theme medial-/trans-Golgi 28:46 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 42 theme medial-/trans-Golgi 28:46 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 42 theme medial-/trans-Golgi 28:46 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 42 theme medial-/trans-Golgi 28:46 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 1 43 theme Yip 279:281 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 43 theme Yip 279:281 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 3 44 theme earlier 677:683 arg1 point 690:694 earlier time point 677:694 earlier time point 677:694 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 8 45 theme BFA 1433:1435 arg1 removal 1422:1428 the removal 1418:1428 the removal of BFA 1418:1435 However, reassembly of the Golgi apparatus after the removal of BFA was markedly delayed by the knockdown of YIPF1 and YIPF2, but not by that of YIPF6. 28286305 0 46 dep reassembly 141:150 arg1 the 131:133 the 131:133 the 131:133 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 4 47 theme YIPF2 931:935 arg1 levels 937:942 YIPF1 and YIPF2 levels 921:942 YIPF1 and YIPF2 levels 921:942 YIPF6 formed a stable complex separately with YIPF1 and YIPF2, and knockdown of YIPF6 reduced YIPF1 and YIPF2 levels. 28286305 1 48 theme domain 283:288 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 48 theme domain 283:288 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 2 49 theme trans-Golgi 518:528 arg1 TGN 539:541 TGN 539:541 TGN 539:541 Immunofluorescence staining revealed that YIPF1, YIPF2, and YIPF6 mainly localize in the medial-/trans-Golgi and also partially in the trans-Golgi network (TGN). 28286305 2 49 theme trans-Golgi 518:528 arg1 network 530:536 the trans-Golgi network 514:536 the trans-Golgi network (TGN) 514:542 Immunofluorescence staining revealed that YIPF1, YIPF2, and YIPF6 mainly localize in the medial-/trans-Golgi and also partially in the trans-Golgi network (TGN). 28286305 5 50 theme Golgi 1071:1075 arg1 apparatus 1077:1085 the Golgi apparatus 1067:1085 the Golgi apparatus 1067:1085 These results suggest that YIPF6 forms complexes with YIPF1 and YIPF2 for their stable expression and localization within the Golgi apparatus. 28286305 3 51 theme medial-/trans-Golgi 619:637 arg1 markers 639:645 medial-/trans-Golgi markers 619:645 medial-/trans-Golgi markers 619:645 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 1 52 theme family 290:295 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 52 theme family 290:295 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 4 53 theme stable 842:847 arg1 complex 849:855 a stable complex 840:855 a stable complex 840:855 YIPF6 formed a stable complex separately with YIPF1 and YIPF2, and knockdown of YIPF6 reduced YIPF1 and YIPF2 levels. 28286305 10 54 theme intracellular 1731:1743 arg1 glycans 1745:1751 intracellular glycans 1731:1751 intracellular glycans 1731:1751 Knockdown of YIPF1 and YIPF2, but not that of YIPF6, also reduced intracellular glycans in HT-29 cells. 28286305 11 55 theme significant 1824:1834 arg1 role 1836:1839 a significant role 1822:1839 a significant role 1822:1839 Thus, we confirmed that YIPF1, YIPF2, and YIPF6 play a significant role in supporting normal glycan synthesis. 28286305 8 56 theme YIPF2 1488:1492 arg1 knockdown 1465:1473 the knockdown 1461:1473 the knockdown of YIPF1 and YIPF2 1461:1492 However, reassembly of the Golgi apparatus after the removal of BFA was markedly delayed by the knockdown of YIPF1 and YIPF2, but not by that of YIPF6. 28286305 0 57 theme Golgi 135:139 arg1 reassembly 141:150 Golgi reassembly 135:150 Golgi reassembly 135:150 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 3 58 theme brefeldin 563:571 arg1 BFA 576:578 BFA 576:578 BFA 576:578 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 3 58 theme brefeldin 563:571 arg1 A 573:573 brefeldin A 563:573 brefeldin A (BFA) 563:579 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 3 59 theme cytoplasmic 730:740 arg1 structures 751:760 cytoplasmic punctate structures 730:760 cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers 730:824 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 0 60 theme trans-Golgi 52:62 arg1 YIPF2 7:11 YIPF2 7:11 YIPF2 7:11 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 60 theme trans-Golgi 52:62 arg1 YIPF6 18:22 YIPF6 18:22 YIPF6 18:22 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 60 theme trans-Golgi 52:62 arg1 YIPF1 0:4 YIPF1 0:4 YIPF1 0:4 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 0 60 theme trans-Golgi 52:62 arg1 proteins 100:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins 28:107 YIPF1, YIPF2, and YIPF6 are medial-/trans-Golgi and trans-Golgi network-localized Yip domain family proteins, which play a role in the Golgi reassembly and glycan synthesis. 28286305 3 61 with treatment 548:556 arg1 BFA 576:578 BFA 576:578 BFA 576:578 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 3 61 with treatment 548:556 arg1 A 573:573 brefeldin A 563:573 brefeldin A (BFA) 563:579 On treatment with brefeldin A (BFA), the homologs co-migrated partly with medial-/trans-Golgi markers and also with a TGN marker in earlier time point, but finally redistributed within cytoplasmic punctate structures that were distinct from medial-/trans-Golgi and the TGN markers. 28286305 1 62 dep homologs 261:268 arg1 homologs 261:268 three uncharacterized mammalian homologs 229:268 three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2 229:351 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 62 dep homologs 261:268 arg1 proteins-YIPF6 297:310 Yip domain family proteins-YIPF6 279:310 Yip domain family proteins-YIPF6 279:310 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 62 dep homologs 261:268 arg1 homolog 315:321 a homolog 313:321 a homolog of Yip1p 313:330 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 1 62 dep homologs 261:268 arg1 YIPF1 337:341 YIPF1 337:341 YIPF1 337:341 In this study, we attempted to explore the function of three uncharacterized mammalian homologs of yeast Yip domain family proteins-YIPF6, a homolog of Yip1p, and YIPF1 and YIPF2, which are homologs of Yif1p. 28286305 10 63 theme YIPF2 1688:1692 arg1 Knockdown 1665:1673 Knockdown 1665:1673 Knockdown of YIPF1 and YIPF2 1665:1692 Knockdown of YIPF1 and YIPF2, but not that of YIPF6, also reduced intracellular glycans in HT-29 cells. 28286305 9 64 theme Golgi 1648:1652 arg1 apparatus 1654:1662 the Golgi apparatus 1644:1662 the Golgi apparatus 1644:1662 These results strongly suggest that free YIPF6 after disassociating with YIPF1 and YIPF2 interferes with the reassembly of the Golgi apparatus. 28323417 5 0 theme line 1089:1092 arg1 slope 1076:1080 the slope 1072:1080 the slope of the line connecting CCS values 1072:1114 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 1 theme 3+ 1147:1148 arg1 ions 1150:1153 3+ ions 1147:1153 3+ ions observed as the glycan antennae 1147:1185 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 2 gly glycoproteins 317:329 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 2 gly glycoproteins 317:329 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 2 gly glycoproteins 317:329 arg1 B 351:351 bovine ribonuclease B 331:351 bovine ribonuclease B 331:351 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 2 gly glycoproteins 317:329 arg1 transferrin 360:370 human transferrin 354:370 human transferrin 354:370 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 2 gly glycoproteins 317:329 arg1 fetuin 380:385 bovine fetuin 373:385 bovine fetuin 373:385 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 3 theme mobility 130:137 arg1 spectrometer 170:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 4 theme set 233:235 arg1 structures 217:226 the gas-phase structures 203:226 the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 203:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 6 5 theme CCS 1391:1393 arg1 increments 1395:1404 the CCS increments 1387:1404 the CCS increments that correspond to changes in glycoform compositions 1387:1457 The results form the basis for a database of CCS values and the CCS increments that correspond to changes in glycoform compositions. 28323417 2 6 theme stepwise 614:621 arg1 removal 623:629 stepwise removal 614:629 stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures 614:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 1 7 theme peptides 452:459 arg1 digestion 281:289 proteolytic digestion 269:289 proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 269:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 8 theme quadrupole 139:148 arg1 spectrometer 170:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 9 gly deglycosylated 437:450 arg1 peptides 452:459 the corresponding deglycosylated peptides 419:459 the corresponding deglycosylated peptides 419:459 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 6 10 theme values 1376:1381 arg1 database 1360:1367 a database 1358:1367 a database of CCS values 1358:1381 The results form the basis for a database of CCS values and the CCS increments that correspond to changes in glycoform compositions. 28323417 1 11 theme human 354:358 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 11 theme human 354:358 arg1 transferrin 360:370 human transferrin 354:370 human transferrin 354:370 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 4 12 theme deglycosylated 916:929 arg1 peptides 931:938 the deglycosylated peptides 912:938 the deglycosylated peptides 912:938 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 2 13 theme nonreducing 671:681 arg1 termini 683:689 the nonreducing termini 667:689 the nonreducing termini of the multiantennary structures 667:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 1 14 theme glycopeptides 240:252 arg1 glycopeptides 240:252 glycopeptides 240:252 glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 240:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 14 theme glycopeptides 240:252 arg1 set 233:235 a set 231:235 a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 231:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 2 15 theme related 533:539 arg1 glycoforms 541:550 closely related glycoforms 525:550 closely related glycoforms 525:550 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 1 16 dep glycoproteins 317:329 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 16 dep glycoproteins 317:329 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 16 dep glycoproteins 317:329 arg1 B 351:351 bovine ribonuclease B 331:351 bovine ribonuclease B 331:351 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 16 dep glycoproteins 317:329 arg1 transferrin 360:370 human transferrin 354:370 human transferrin 354:370 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 16 dep glycoproteins 317:329 arg1 fetuin 380:385 bovine fetuin 373:385 bovine fetuin 373:385 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 2 17 theme units 656:660 arg1 removal 623:629 stepwise removal 614:629 stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures 614:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 4 18 gly glycopeptides 897:909 arg2 glycopeptides 897:909 individual N-linked glycopeptides 877:909 individual N-linked glycopeptides 877:909 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 4 19 dep released 949:956 arg1 deutero-reduced 959:973 deutero-reduced 959:973 deutero-reduced 959:973 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 1 20 theme bovine 373:378 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 20 theme bovine 373:378 arg1 fetuin 380:385 bovine fetuin 373:385 bovine fetuin 373:385 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 2 21 theme saccharide 645:654 arg1 units 656:660 individual saccharide units 634:660 individual saccharide units 634:660 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 1 22 theme glycans 470:476 arg1 digestion 281:289 proteolytic digestion 269:289 proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 269:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 23 theme given 1022:1026 arg1 glycopeptide 1028:1039 a given glycopeptide or set 1020:1046 glycopeptide 1028:1039 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 24 theme mass 165:168 arg1 spectrometer 170:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 25 theme peptide 1285:1291 arg1 length 1293:1298 the peptide length 1281:1298 the peptide length 1281:1298 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 2 26 theme individual 634:643 arg1 units 656:660 individual saccharide units 634:660 individual saccharide units 634:660 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 2 27 theme multiantennary 698:711 arg1 structures 713:722 the multiantennary structures 694:722 the multiantennary structures 694:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 4 28 theme peptides 931:938 arg1 glycoforms 863:872 the glycoforms 859:872 the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans 859:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 3 29 theme Collision 725:733 arg1 CCS 751:753 CCS 751:753 CCS 751:753 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 3 29 theme Collision 725:733 arg1 sections 741:748 Collision cross sections 725:748 Collision cross sections (CCS) 725:754 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 3 29 theme Collision 725:733 arg1 function 789:796 a function 787:796 a function of mass-to-charge ratio 787:820 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 4 30 theme permethylated 975:987 arg1 glycans 989:995 the released, deutero-reduced permethylated glycans 945:995 the released, deutero-reduced permethylated glycans 945:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 5 31 theme exoglycosidase 1214:1227 arg1 treatments 1229:1238 stepwise exoglycosidase treatments 1205:1238 stepwise exoglycosidase treatments 1205:1238 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 4 32 theme individual 877:886 arg1 glycopeptides 897:909 individual N-linked glycopeptides 877:909 individual N-linked glycopeptides 877:909 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 3 33 theme mass-to-charge 801:814 arg1 ratio 816:820 mass-to-charge ratio 801:820 mass-to-charge ratio 801:820 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 4 34 gly deglycosylated 916:929 arg1 peptides 931:938 the deglycosylated peptides 912:938 the deglycosylated peptides 912:938 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 0 35 theme Database 18:25 arg1 Construction 0:11 Construction 0:11 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides 0:100 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS. 28323417 1 36 theme proteolytic 269:279 arg1 digestion 281:289 proteolytic digestion 269:289 proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 269:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 37 gly glycopeptides 240:252 arg2 glycopeptides 240:252 glycopeptides 240:252 glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 240:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 4 38 theme glycopeptides 897:909 arg1 glycoforms 863:872 the glycoforms 859:872 the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans 859:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 4 39 gly glycoforms 863:872 arg1 glycans 989:995 the released, deutero-reduced permethylated glycans 945:995 the released, deutero-reduced permethylated glycans 945:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 4 39 gly glycoforms 863:872 arg1 peptides 931:938 the deglycosylated peptides 912:938 the deglycosylated peptides 912:938 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 4 39 gly glycoforms 863:872 arg1 glycopeptides 897:909 individual N-linked glycopeptides 877:909 individual N-linked glycopeptides 877:909 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 4 40 theme N-linked 888:895 arg1 glycopeptides 897:909 individual N-linked glycopeptides 877:909 individual N-linked glycopeptides 877:909 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 6 41 theme glycoform 1436:1444 arg1 compositions 1446:1457 glycoform compositions 1436:1457 glycoform compositions 1436:1457 The results form the basis for a database of CCS values and the CCS increments that correspond to changes in glycoform compositions. 28323417 4 42 theme released 949:956 arg1 glycans 989:995 the released, deutero-reduced permethylated glycans 945:995 the released, deutero-reduced permethylated glycans 945:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 6 43 theme CCS 1372:1374 arg1 values 1376:1381 CCS values 1372:1381 CCS values 1372:1381 The results form the basis for a database of CCS values and the CCS increments that correspond to changes in glycoform compositions. 28323417 5 44 theme glycan 1171:1176 arg1 antennae 1178:1185 the glycan antennae 1167:1185 the glycan antennae 1167:1185 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 0 45 theme Collision 30:38 arg1 Section 46:52 Collision Cross Section 30:52 Collision Cross Section Values 30:59 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS. 28323417 5 46 theme stepwise 1205:1212 arg1 treatments 1229:1238 stepwise exoglycosidase treatments 1205:1238 stepwise exoglycosidase treatments 1205:1238 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 4 47 theme glycans 989:995 arg1 glycoforms 863:872 the glycoforms 859:872 the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans 859:995 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 1 48 theme human 391:395 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 48 theme human 391:395 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 49 theme derivatized 1051:1061 arg1 glycans 1063:1069 derivatized glycans 1051:1069 derivatized glycans 1051:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 50 gly glycopeptide 1028:1039 arg2 glycopeptide 1028:1039 a given glycopeptide or set 1020:1046 glycopeptide 1028:1039 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 50 gly glycopeptide 1028:1039 arg1 glycans 1063:1069 derivatized glycans 1051:1069 derivatized glycans 1051:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 51 theme α1-acid 397:403 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 51 theme α1-acid 397:403 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 52 theme glycans 1063:1069 arg1 set 1044:1046 a given glycopeptide or set 1020:1046 set 1044:1046 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 52 theme glycans 1063:1069 arg1 glycans 1063:1069 derivatized glycans 1051:1069 derivatized glycans 1051:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 52 theme glycans 1063:1069 arg1 glycopeptide 1028:1039 a given glycopeptide or set 1020:1046 glycopeptide 1028:1039 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 53 theme endoglycosidase 494:508 arg1 F 517:517 the endoglycosidase PNGase F 490:517 the endoglycosidase PNGase F 490:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 6 54 from changes 1425:1431 arg1 compositions 1446:1457 glycoform compositions 1436:1457 glycoform compositions 1436:1457 The results form the basis for a database of CCS values and the CCS increments that correspond to changes in glycoform compositions. 28323417 0 55 theme Section 46:52 arg1 Values 54:59 Collision Cross Section Values 30:59 Collision Cross Section Values 30:59 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS. 28323417 1 56 theme well-characterized 298:315 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 56 theme well-characterized 298:315 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 56 theme well-characterized 298:315 arg1 B 351:351 bovine ribonuclease B 331:351 bovine ribonuclease B 331:351 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 56 theme well-characterized 298:315 arg1 transferrin 360:370 human transferrin 354:370 human transferrin 354:370 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 56 theme well-characterized 298:315 arg1 fetuin 380:385 bovine fetuin 373:385 bovine fetuin 373:385 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 57 gly glycoforms 1006:1015 arg1 set 1044:1046 a given glycopeptide or set 1020:1046 set 1044:1046 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 57 gly glycoforms 1006:1015 arg1 glycans 1063:1069 derivatized glycans 1051:1069 derivatized glycans 1051:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 5 57 gly glycoforms 1006:1015 arg1 glycopeptide 1028:1039 a given glycopeptide or set 1020:1046 glycopeptide 1028:1039 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 58 theme PNGase 510:515 arg1 F 517:517 the endoglycosidase PNGase F 490:517 the endoglycosidase PNGase F 490:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 0 59 theme Cross 40:44 arg1 Section 46:52 Collision Cross Section 30:52 Collision Cross Section Values 30:59 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS. 28323417 1 60 theme glycoproteins 317:329 arg1 digestion 281:289 proteolytic digestion 269:289 proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 269:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 3 61 theme cross 735:739 arg1 CCS 751:753 CCS 751:753 CCS 751:753 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 3 61 theme cross 735:739 arg1 sections 741:748 Collision cross sections 725:748 Collision cross sections (CCS) 725:754 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 3 61 theme cross 735:739 arg1 function 789:796 a function 787:796 a function of mass-to-charge ratio 787:820 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 1 62 used used 187:190 arg2 spectrometer 170:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 63 theme bovine 331:336 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 63 theme bovine 331:336 arg1 B 351:351 bovine ribonuclease B 331:351 bovine ribonuclease B 331:351 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 4 64 link N-linked 888:895 arg1 glycopeptides 897:909 individual N-linked glycopeptides 877:909 individual N-linked glycopeptides 877:909 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 2 65 theme structures 713:722 arg1 termini 683:689 the nonreducing termini 667:689 the nonreducing termini of the multiantennary structures 667:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 5 66 theme glycopeptide 1028:1039 arg1 glycoforms 1006:1015 the glycoforms 1002:1015 the glycoforms of a given glycopeptide or set of derivatized glycans 1002:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 2 67 from termini 683:689 arg1 removal 623:629 stepwise removal 614:629 stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures 614:722 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 28323417 1 68 theme gas-phase 207:215 arg1 structures 217:226 the gas-phase structures 203:226 the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F 203:517 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 69 theme corresponding 423:435 arg1 peptides 452:459 the corresponding deglycosylated peptides 419:459 the corresponding deglycosylated peptides 419:459 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 70 theme CCS 1105:1107 arg1 values 1109:1114 CCS values 1105:1114 CCS values 1105:1114 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 71 gly glycoprotein 405:416 arg1 glycoprotein 405:416 human α1-acid glycoprotein 391:416 human α1-acid glycoprotein 391:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 71 gly glycoprotein 405:416 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 72 dep remained 1116:1123 arg1 shortened 1192:1200 shortened 1192:1200 were shortened by stepwise exoglycosidase treatments 1187:1238 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 1 73 theme deglycosylated 437:450 arg1 peptides 452:459 the corresponding deglycosylated peptides 419:459 the corresponding deglycosylated peptides 419:459 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 5 74 theme set 1044:1046 arg1 glycoforms 1006:1015 the glycoforms 1002:1015 the glycoforms of a given glycopeptide or set of derivatized glycans 1002:1069 For the glycoforms of a given glycopeptide or set of derivatized glycans, the slope of the line connecting CCS values remained similar for the [M+3H]3+ ions observed as the glycan antennae were shortened by stepwise exoglycosidase treatments; this trend was consistent regardless of the peptide length or the saccharide removed. 28323417 3 75 theme ratio 816:820 arg1 function 789:796 a function 787:796 a function of mass-to-charge ratio 787:820 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 3 75 theme ratio 816:820 arg1 sections 741:748 Collision cross sections 725:748 Collision cross sections (CCS) 725:754 Collision cross sections (CCS) were calculated and plotted as a function of mass-to-charge ratio. 28323417 0 76 theme Values 54:59 arg1 Database 18:25 a Database 16:25 a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides 16:100 Construction of a Database of Collision Cross Section Values for Glycopeptides, Glycans, and Peptides Determined by IM-MS. 28323417 1 77 theme ribonuclease 338:349 arg1 glycoproteins 317:329 the well-characterized glycoproteins 294:329 the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein 294:416 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 1 77 theme ribonuclease 338:349 arg1 B 351:351 bovine ribonuclease B 331:351 bovine ribonuclease B 331:351 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 4 78 theme Linear 823:828 arg1 trendlines 830:839 Linear trendlines 823:839 Linear trendlines 823:839 Linear trendlines were observed for the glycoforms of individual N-linked glycopeptides, the deglycosylated peptides, and the released, deutero-reduced permethylated glycans. 28323417 1 79 theme ion 126:128 arg1 spectrometer 170:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer 123:181 An ion mobility quadrupole time-of-flight mass spectrometer was used to examine the gas-phase structures of a set of glycopeptides resulting from proteolytic digestion of the well-characterized glycoproteins bovine ribonuclease B, human transferrin, bovine fetuin and human α1-acid glycoprotein, the corresponding deglycosylated peptides, and the glycans released by the endoglycosidase PNGase F. 28323417 2 80 used used 598:601 arg2 exoglycosidases 577:591 exoglycosidases 577:591 exoglycosidases 577:591 When closely related glycoforms did not occur naturally, exoglycosidases were used to achieve stepwise removal of individual saccharide units from the nonreducing termini of the multiantennary structures. 27313150 7 0 theme important 1574:1582 arg1 step 1584:1587 an important step 1571:1587 an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies 1571:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 1 theme due 282:284 arg1 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 1 theme due 282:284 arg1 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 3 2 theme tumor-targeting 573:587 arg1 H10 613:615 the tumor-targeting monoclonal antibody mAb H10 569:615 the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 569:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 3 with H10 693:695 arg1 rhizogenes 705:714 A. rhizogenes 702:714 A. rhizogenes 702:714 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 7 4 theme mAb 1498:1500 arg1 H10 1502:1504 the tumor-targeting mAb H10 1478:1504 the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1478:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 5 attach derived 152:158 arg2 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 5 attach derived 152:158 arg2 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 5 attach derived 152:158 arg1 transformation 190:203 Agrobacterium rhizogenes transformation 165:203 Agrobacterium rhizogenes transformation of plant tissues 165:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 7 6 with co-infection 1325:1336 arg1 rhizogenes 1389:1398 recombinant A. rhizogenes 1374:1398 recombinant A. rhizogenes 1374:1398 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 7 theme Agrobacterium 165:177 arg1 transformation 190:203 Agrobacterium rhizogenes transformation 165:203 Agrobacterium rhizogenes transformation of plant tissues 165:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 7 8 theme glycosylation 1530:1542 arg1 profile 1544:1550 a human-compatible glycosylation profile 1511:1550 a human-compatible glycosylation profile 1511:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 4 9 theme similar 934:940 arg1 yields 942:947 similar yields 934:947 similar yields (2-3 mg/L) 934:958 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 4 9 theme similar 934:940 arg1 mg/L 954:957 2-3 mg/L 950:957 2-3 mg/L 950:957 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 7 10 theme cultures 1458:1465 arg1 generation 1434:1443 the generation 1430:1443 the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1430:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 5 11 theme N. 1091:1092 arg1 H10 1114:1116 N. tabacum-derived mAb H10 1091:1116 N. tabacum-derived mAb H10 1091:1116 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 3 12 theme antibody 600:607 arg1 H10 613:615 the tumor-targeting monoclonal antibody mAb H10 569:615 the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 569:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 13 dep infecting 643:651 arg1 either 633:638 either 633:638 either 633:638 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 7 14 theme stable 1448:1453 arg1 cultures 1458:1465 stable HR cultures 1448:1465 stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1448:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 3 15 theme mAb 806:808 arg1 heavy 814:818 mAb H10 heavy 806:818 mAb H10 heavy 806:818 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 5 16 theme mAb 1110:1112 arg1 H10 1114:1116 N. tabacum-derived mAb H10 1091:1116 N. tabacum-derived mAb H10 1091:1116 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 5 17 theme plant-typical 1054:1066 arg1 structures 1076:1085 plant-typical complex structures 1054:1085 plant-typical complex structures 1054:1085 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 1 18 theme Hairy 127:131 arg1 root 133:136 Hairy root 127:136 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 18 theme Hairy 127:131 arg1 HR 139:140 HR 139:140 HR 139:140 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 7 19 theme antibodies 1689:1698 arg1 production 1639:1648 the production 1635:1648 the production of 'next generation' human therapeutic antibodies 1635:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 6 20 theme binding 1259:1265 arg1 activities 1267:1276 comparable antigen binding activities 1240:1276 comparable antigen binding activities 1240:1276 Both antibody glyco-formats exhibited comparable antigen binding activities. 27313150 3 21 theme light 824:828 arg1 chain 830:834 light chain 824:834 light chain 824:834 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 5 22 theme structures 1076:1085 arg1 presence 1042:1049 the presence 1038:1049 the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart 1038:1199 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 3 23 theme transgenic 655:664 arg1 line 677:680 a transgenic N. tabacum line 653:680 a transgenic N. tabacum line expressing H10 with A. rhizogenes 653:714 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 2 24 used used 474:477 arg2 HRs 400:402 HRs 400:402 HRs descending from transgenic Nicotiana tabacum plants 400:454 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 7 25 theme human 1671:1675 arg1 antibodies 1689:1698 'next generation' human therapeutic antibodies 1653:1698 'next generation' human therapeutic antibodies 1653:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 2 26 theme plant-type 530:539 arg1 glycans 541:547 plant-type glycans 530:547 plant-type glycans 530:547 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 6 27 theme comparable 1240:1249 arg1 activities 1267:1276 comparable antigen binding activities 1240:1276 comparable antigen binding activities 1240:1276 Both antibody glyco-formats exhibited comparable antigen binding activities. 27313150 3 28 from H10 613:615 arg1 infecting 643:651 infecting 643:651 infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 643:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 28 from H10 613:615 arg1 HRs 620:622 HRs 620:622 HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 620:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 29 theme heavy 814:818 arg1 cDNAs 836:840 mAb H10 heavy and light chain cDNAs 806:840 mAb H10 heavy and light chain cDNAs 806:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 0 30 theme glycosylation 66:78 arg1 profile 80:86 a human-compatible glycosylation profile 47:86 a human-compatible glycosylation profile in N. benthamiana hairy root cultures 47:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 7 31 theme generation 1659:1668 arg1 antibodies 1689:1698 'next generation' human therapeutic antibodies 1653:1698 'next generation' human therapeutic antibodies 1653:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 2 32 theme mAbs 520:523 arg1 production 487:496 the production 483:496 the production of several functional mAbs with plant-type glycans 483:547 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 1 33 theme culture 367:373 arg1 medium 375:380 an otherwise largely protein free culture medium 333:380 an otherwise largely protein free culture medium 333:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 2 34 with mAbs 520:523 arg1 glycans 541:547 plant-type glycans 530:547 plant-type glycans 530:547 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 3 35 contain carrying 797:804 arg2 cDNAs 836:840 mAb H10 heavy and light chain cDNAs 806:840 mAb H10 heavy and light chain cDNAs 806:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 35 contain carrying 797:804 arg1 rhizogenes 786:795 A. rhizogenes 783:795 recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 771:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 0 36 theme benthamiana 94:104 arg1 cultures 117:124 N. benthamiana hairy root cultures 91:124 N. benthamiana hairy root cultures 91:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 0 37 theme root 112:115 arg1 cultures 117:124 N. benthamiana hairy root cultures 91:124 N. benthamiana hairy root cultures 91:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 5 38 theme GnGn 1125:1128 arg1 structures 1130:1139 GnGn structures 1125:1139 GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart 1125:1199 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 5 39 theme antibodies 989:998 arg1 profiles 977:984 N-glycosylation profiles 961:984 N-glycosylation profiles of antibodies purified from HR supernatant 961:1027 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 7 40 theme efficient 1406:1414 arg1 co-infection 1325:1336 the co-infection 1321:1336 the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes 1321:1398 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 7 40 theme efficient 1406:1414 arg1 procedure 1416:1424 an efficient procedure 1403:1424 an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1403:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 41 theme recombinant 309:319 arg1 proteins 321:328 recombinant proteins 309:328 recombinant proteins 309:328 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 7 42 theme cultures 1622:1629 arg1 exploitation 1601:1612 the exploitation 1597:1612 the exploitation of root cultures 1597:1629 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 7 43 theme recombinant 1374:1384 arg1 rhizogenes 1389:1398 recombinant A. rhizogenes 1374:1398 recombinant A. rhizogenes 1374:1398 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 2 44 theme several 501:507 arg1 mAbs 520:523 several functional mAbs 501:523 several functional mAbs with plant-type glycans 501:547 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 2 45 theme Nicotiana 431:439 arg1 plants 449:454 transgenic Nicotiana tabacum plants 420:454 transgenic Nicotiana tabacum plants 420:454 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 0 46 theme antibody 33:40 arg1 Production 0:9 Production 0:9 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 0:125 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 7 47 with H10 1502:1504 arg1 profile 1544:1550 a human-compatible glycosylation profile 1511:1550 a human-compatible glycosylation profile 1511:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 5 48 link ΔXTFT-derived 1175:1187 arg1 counterpart 1189:1199 the ΔXTFT-derived counterpart 1171:1199 the ΔXTFT-derived counterpart 1171:1199 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 7 49 theme benthamiana 1357:1367 arg1 co-infection 1325:1336 the co-infection 1321:1336 the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes 1321:1398 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 7 49 theme benthamiana 1357:1367 arg1 procedure 1416:1424 an efficient procedure 1403:1424 an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1403:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 3 50 theme tabacum 669:675 arg1 line 677:680 a transgenic N. tabacum line 653:680 a transgenic N. tabacum line expressing H10 with A. rhizogenes 653:714 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 4 51 theme HR 852:853 arg1 clones 855:860 Selected HR clones 843:860 Selected HR clones derived from both plants 843:885 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 3 52 theme benthamiana 741:751 arg1 line 753:756 a glyco-engineered N. benthamiana line 719:756 a glyco-engineered N. benthamiana line (ΔXTFT) 719:764 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 52 theme benthamiana 741:751 arg1 ΔXTFT 759:763 ΔXTFT 759:763 ΔXTFT 759:763 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 5 53 theme ΔXTFT-derived 1175:1187 arg1 counterpart 1189:1199 the ΔXTFT-derived counterpart 1171:1199 the ΔXTFT-derived counterpart 1171:1199 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 7 54 theme ΔXTFT 1341:1345 arg1 benthamiana 1357:1367 ΔXTFT Nicotiana benthamiana 1341:1367 ΔXTFT Nicotiana benthamiana 1341:1367 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 0 55 with Production 0:9 arg1 profile 80:86 a human-compatible glycosylation profile 47:86 a human-compatible glycosylation profile in N. benthamiana hairy root cultures 47:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 1 56 theme biotechnological 242:257 arg1 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 56 theme biotechnological 242:257 arg1 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 3 57 theme N. 738:739 arg1 line 753:756 a glyco-engineered N. benthamiana line 719:756 a glyco-engineered N. benthamiana line (ΔXTFT) 719:764 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 57 theme N. 738:739 arg1 ΔXTFT 759:763 ΔXTFT 759:763 ΔXTFT 759:763 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 4 58 theme mAb 899:901 arg1 H10 903:905 mAb H10 899:905 mAb H10 899:905 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 6 59 theme antibody 1207:1214 arg1 glyco-formats 1216:1228 Both antibody glyco-formats 1202:1228 Both antibody glyco-formats 1202:1228 Both antibody glyco-formats exhibited comparable antigen binding activities. 27313150 7 60 theme human-compatible 1513:1528 arg1 profile 1544:1550 a human-compatible glycosylation profile 1511:1550 a human-compatible glycosylation profile 1511:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 61 from accumulation 293:304 arg1 medium 375:380 an otherwise largely protein free culture medium 333:380 an otherwise largely protein free culture medium 333:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 3 62 theme mAb 609:611 arg1 H10 613:615 the tumor-targeting monoclonal antibody mAb H10 569:615 the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 569:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 63 dep obtained 624:631 arg1 infecting 643:651 infecting 643:651 infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 643:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 63 dep obtained 624:631 arg1 HRs 620:622 HRs 620:622 HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 620:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 7 64 theme tumor-targeting 1482:1496 arg1 H10 1502:1504 the tumor-targeting mAb H10 1478:1504 the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1478:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 1 65 dep Agrobacterium 165:177 arg1 rhizogenes 179:188 rhizogenes 179:188 rhizogenes 179:188 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 3 66 theme monoclonal 589:598 arg1 H10 613:615 the tumor-targeting monoclonal antibody mAb H10 569:615 the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs 569:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 5 67 theme tabacum-derived 1094:1108 arg1 H10 1114:1116 N. tabacum-derived mAb H10 1091:1116 N. tabacum-derived mAb H10 1091:1116 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 1 68 theme tissues 214:220 arg1 transformation 190:203 Agrobacterium rhizogenes transformation 165:203 Agrobacterium rhizogenes transformation of plant tissues 165:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 0 69 from profile 80:86 arg1 cultures 117:124 N. benthamiana hairy root cultures 91:124 N. benthamiana hairy root cultures 91:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 7 70 theme HR 1455:1456 arg1 cultures 1458:1465 stable HR cultures 1448:1465 stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile 1448:1550 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 3 71 theme H10 810:812 arg1 heavy 814:818 mAb H10 heavy 806:818 mAb H10 heavy 806:818 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 7 72 theme therapeutic 1677:1687 arg1 antibodies 1689:1698 'next generation' human therapeutic antibodies 1653:1698 'next generation' human therapeutic antibodies 1653:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 5 73 theme complex 1068:1074 arg1 structures 1076:1085 plant-typical complex structures 1054:1085 plant-typical complex structures 1054:1085 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 1 74 theme root 133:136 arg1 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 74 theme root 133:136 arg1 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 4 75 theme culture 914:920 arg1 medium 922:927 the culture medium 910:927 the culture medium 910:927 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 6 76 theme antigen 1251:1257 arg1 activities 1267:1276 comparable antigen binding activities 1240:1276 comparable antigen binding activities 1240:1276 Both antibody glyco-formats exhibited comparable antigen binding activities. 27313150 3 77 theme chain 830:834 arg1 cDNAs 836:840 mAb H10 heavy and light chain cDNAs 806:840 mAb H10 heavy and light chain cDNAs 806:840 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 7 78 theme next 1654:1657 arg1 generation 1659:1668 'next generation' 1653:1669 'next generation' human therapeutic antibodies 1653:1698 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 5 79 gly N-glycosylation 961:975 arg1 antibodies 989:998 antibodies 989:998 antibodies purified from HR supernatant 989:1027 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 0 80 theme N. 91:92 arg1 cultures 117:124 N. benthamiana hairy root cultures 91:124 N. benthamiana hairy root cultures 91:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 1 81 theme free 362:365 arg1 medium 375:380 an otherwise largely protein free culture medium 333:380 an otherwise largely protein free culture medium 333:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 4 82 attach derived 862:868 arg2 clones 855:860 Selected HR clones 843:860 Selected HR clones derived from both plants 843:885 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 4 82 attach derived 862:868 arg1 plants 880:885 both plants 875:885 both plants 875:885 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 2 83 theme functional 509:518 arg1 mAbs 520:523 several functional mAbs 501:523 several functional mAbs with plant-type glycans 501:547 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 0 84 theme hairy 106:110 arg1 cultures 117:124 N. benthamiana hairy root cultures 91:124 N. benthamiana hairy root cultures 91:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 5 85 theme N-glycosylation 961:975 arg1 profiles 977:984 N-glycosylation profiles 961:984 N-glycosylation profiles of antibodies purified from HR supernatant 961:1027 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 5 86 link tabacum-derived 1094:1108 arg1 H10 1114:1116 N. tabacum-derived mAb H10 1091:1116 N. tabacum-derived mAb H10 1091:1116 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 1 87 theme plant 208:212 arg1 tissues 214:220 plant tissues 208:220 plant tissues 208:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 5 88 theme structures 1130:1139 arg1 presence 1042:1049 the presence 1038:1049 the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart 1038:1199 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 7 89 theme A. 1386:1387 arg1 rhizogenes 1389:1398 recombinant A. rhizogenes 1374:1398 recombinant A. rhizogenes 1374:1398 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 0 90 theme tumour-targeting 16:31 arg1 antibody 33:40 a tumour-targeting antibody 14:40 a tumour-targeting antibody 14:40 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 2 91 theme tabacum 441:447 arg1 plants 449:454 transgenic Nicotiana tabacum plants 420:454 transgenic Nicotiana tabacum plants 420:454 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 1 92 theme proteins 321:328 arg1 accumulation 293:304 the accumulation 289:304 the accumulation of recombinant proteins in an otherwise largely protein free culture medium 289:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 7 93 theme Nicotiana 1347:1355 arg1 benthamiana 1357:1367 ΔXTFT Nicotiana benthamiana 1341:1367 ΔXTFT Nicotiana benthamiana 1341:1367 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 3 94 theme N. 666:667 arg1 line 677:680 a transgenic N. tabacum line 653:680 a transgenic N. tabacum line expressing H10 with A. rhizogenes 653:714 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 2 95 theme transgenic 420:429 arg1 plants 449:454 transgenic Nicotiana tabacum plants 420:454 transgenic Nicotiana tabacum plants 420:454 In this context, HRs descending from transgenic Nicotiana tabacum plants were successfully used for the production of several functional mAbs with plant-type glycans. 27313150 0 96 theme human-compatible 49:64 arg1 profile 80:86 a human-compatible glycosylation profile 47:86 a human-compatible glycosylation profile in N. benthamiana hairy root cultures 47:124 Production of a tumour-targeting antibody with a human-compatible glycosylation profile in N. benthamiana hairy root cultures. 27313150 7 97 theme root 1617:1620 arg1 cultures 1622:1629 root cultures 1617:1629 root cultures 1617:1629 Collectively, these data demonstrate that the co-infection of ΔXTFT Nicotiana benthamiana with recombinant A. rhizogenes is an efficient procedure for the generation of stable HR cultures expressing the tumor-targeting mAb H10 with a human-compatible glycosylation profile, thus representing an important step towards the exploitation of root cultures for the production of 'next generation' human therapeutic antibodies. 27313150 4 98 theme Selected 843:850 arg1 clones 855:860 Selected HR clones 843:860 Selected HR clones derived from both plants 843:885 Selected HR clones derived from both plants accumulated mAb H10 in the culture medium with similar yields (2-3 mg/L). 27313150 1 99 theme advantageous 229:240 arg1 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 99 theme advantageous 229:240 arg1 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 5 100 theme HR 1014:1015 arg1 supernatant 1017:1027 HR supernatant 1014:1027 HR supernatant 1014:1027 N-glycosylation profiles of antibodies purified from HR supernatant revealed the presence of plant-typical complex structures for N. tabacum-derived mAb H10 and of GnGn structures lacking xylose and fucose for the ΔXTFT-derived counterpart. 27313150 3 101 theme glyco-engineered 721:736 arg1 line 753:756 a glyco-engineered N. benthamiana line 719:756 a glyco-engineered N. benthamiana line (ΔXTFT) 719:764 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 3 101 theme glyco-engineered 721:736 arg1 ΔXTFT 759:763 ΔXTFT 759:763 ΔXTFT 759:763 Here, we expressed the tumor-targeting monoclonal antibody mAb H10 in HRs obtained either by infecting a transgenic N. tabacum line expressing H10 with A. rhizogenes or a glyco-engineered N. benthamiana line (ΔXTFT) with recombinant A. rhizogenes carrying mAb H10 heavy and light chain cDNAs. 27313150 1 102 theme manufacturing 259:271 arg1 cultures 143:150 Hairy root (HR) cultures 127:150 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues 127:220 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 27313150 1 102 theme manufacturing 259:271 arg1 platform 273:280 an advantageous biotechnological manufacturing platform 226:280 an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium 226:380 Hairy root (HR) cultures derived from Agrobacterium rhizogenes transformation of plant tissues are an advantageous biotechnological manufacturing platform due to the accumulation of recombinant proteins in an otherwise largely protein free culture medium. 26432054 3 0 theme oxidative 689:697 arg1 burst 699:703 an oxidative burst 686:703 an oxidative burst 686:703 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 2 1 theme diverse 348:354 arg1 oligosaccharides 365:380 structurally diverse chitosan oligosaccharides 335:380 structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) 335:419 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 5 2 theme PA 1105:1106 arg1 values 1108:1113 defined DA and PA values 1090:1113 defined DA and PA values 1090:1113 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 0 3 theme Serratia 101:108 arg1 proteamaculans 110:123 Serratia proteamaculans 101:123 Serratia proteamaculans 101:123 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 2 4 theme W114A 505:509 arg1 mutant 511:516 the W114A mutant 501:516 the W114A mutant which lacks transglycosylase activity 501:554 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 1 5 theme acetylation 279:289 arg1 degree 238:243 the degree 234:243 the degree of polymerization (DP) 234:266 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 5 theme acetylation 279:289 arg1 pattern 300:306 pattern 300:306 pattern of acetylation (PA) 300:326 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 5 theme acetylation 279:289 arg1 degree 269:274 degree 269:274 degree of acetylation (DA) 269:294 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 3 6 with mixtures 583:590 arg1 ranges 639:644 DA ranges 636:644 DA ranges 636:644 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 3 6 with mixtures 583:590 arg1 DP 629:630 specific DP 620:630 specific DP 620:630 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 3 7 theme rice 708:711 arg1 cultures 729:736 rice cell suspension cultures 708:736 rice cell suspension cultures 708:736 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 2 8 dep proteamaculans 451:464 arg1 SpChiD 489:494 SpChiD 489:494 SpChiD 489:494 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 2 8 dep proteamaculans 451:464 arg1 D 486:486 chitinase D 476:486 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 4 9 theme crude 743:747 arg1 mixtures 749:756 The crude mixtures 739:756 The crude mixtures 739:756 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 4 9 theme crude 743:747 arg1 active 768:773 active 768:773 active 768:773 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 3 10 theme cell 713:716 arg1 cultures 729:736 rice cell suspension cultures 708:736 rice cell suspension cultures 708:736 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 4 11 theme purified 821:828 arg1 fractions 830:838 the purified fractions 817:838 the purified fractions 817:838 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 4 11 theme purified 821:828 arg1 active 850:855 active 850:855 active 850:855 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 5 12 with oligosaccharides 1068:1083 arg1 values 1108:1113 defined DA and PA values 1090:1113 defined DA and PA values 1090:1113 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 2 13 theme chitinase 476:484 arg1 SpChiD 489:494 SpChiD 489:494 SpChiD 489:494 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 2 13 theme chitinase 476:484 arg1 D 486:486 chitinase D 476:486 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 2 14 theme chitosan 387:394 arg1 polymers 396:403 chitosan polymers 387:403 chitosan polymers (DA=35% or 61%) 387:419 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 3 15 theme crude 561:565 arg1 mixtures 583:590 The crude oligosaccharide mixtures 557:590 The crude oligosaccharide mixtures 557:590 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 0 16 theme chitosan 24:31 arg1 Production 0:9 Production 0:9 Production of bioactive chitosan 0:31 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 2 17 theme transglycosylase 530:545 arg1 activity 547:554 transglycosylase activity 530:554 transglycosylase activity 530:554 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 1 18 theme biological 130:139 arg1 activities 141:150 The biological activities 126:150 The biological activities of chitosan and its oligosaccharides 126:187 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 0 19 theme bioactive 14:22 arg1 chitosan 24:31 bioactive chitosan 14:31 bioactive chitosan 14:31 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 3 20 theme suspension 718:727 arg1 cultures 729:736 rice cell suspension cultures 708:736 rice cell suspension cultures 708:736 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 0 21 from proteamaculans 110:123 arg1 chitinase-D 84:94 the hypertransglycosylating chitinase-D 56:94 the hypertransglycosylating chitinase-D from Serratia proteamaculans 56:123 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 2 22 theme wild-type 466:474 arg1 proteamaculans 451:464 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 5 23 gly transglycosylation 915:932 arg1 presence 965:972 the presence 961:972 the presence of fully-deacetylated oligosaccharides 961:1011 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 5 24 theme defined 1090:1096 arg1 values 1108:1113 defined DA and PA values 1090:1113 defined DA and PA values 1090:1113 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 4 25 theme W114A 796:800 arg1 mutant 802:807 the W114A mutant 792:807 the W114A mutant 792:807 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 5 26 theme DA 1098:1099 arg1 values 1108:1113 defined DA and PA values 1090:1113 defined DA and PA values 1090:1113 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 3 27 theme purified 596:603 arg1 fractions 605:613 purified fractions 596:613 purified fractions with specific DP and DA ranges 596:644 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 1 28 theme polymerization 248:261 arg1 degree 238:243 the degree 234:243 the degree of polymerization (DP) 234:266 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 28 theme polymerization 248:261 arg1 pattern 300:306 pattern 300:306 pattern of acetylation (PA) 300:326 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 28 theme polymerization 248:261 arg1 degree 269:274 degree 269:274 degree of acetylation (DA) 269:294 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 2 29 theme Serratia 442:449 arg1 proteamaculans 451:464 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Serratia proteamaculans wild-type chitinase D (SpChiD) 442:495 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 1 30 theme acetylation 311:321 arg1 degree 238:243 the degree 234:243 the degree of polymerization (DP) 234:266 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 30 theme acetylation 311:321 arg1 pattern 300:306 pattern 300:306 pattern of acetylation (PA) 300:326 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 1 30 theme acetylation 311:321 arg1 degree 269:274 degree 269:274 degree of acetylation (DA) 269:294 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 4 31 theme wild-type 874:882 arg1 SpChiD 884:889 wild-type SpChiD 874:889 wild-type SpChiD 874:889 The crude mixtures were more active when produced by the W114A mutant whereas the purified fractions were more active when produced by wild-type SpChiD. 26432054 3 32 theme DA 636:637 arg1 ranges 639:644 DA ranges 636:644 DA ranges 636:644 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 3 33 theme oligosaccharide 567:581 arg1 mixtures 583:590 The crude oligosaccharide mixtures 557:590 The crude oligosaccharide mixtures 557:590 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 1 34 theme chitosan 155:162 arg1 activities 141:150 The biological activities 126:150 The biological activities of chitosan and its oligosaccharides 126:187 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 0 35 gly hypertransglycosylating 60:82 arg0 chitinase-D 84:94 the hypertransglycosylating chitinase-D 56:94 the hypertransglycosylating chitinase-D from Serratia proteamaculans 56:123 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 5 36 theme fully-deacetylated 977:994 arg1 oligosaccharides 996:1011 fully-deacetylated oligosaccharides 977:1011 fully-deacetylated oligosaccharides 977:1011 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 3 37 theme specific 620:627 arg1 DP 629:630 specific DP 620:630 specific DP 620:630 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 2 38 from polymers 396:403 arg1 oligosaccharides 365:380 structurally diverse chitosan oligosaccharides 335:380 structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) 335:419 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 5 39 theme oligosaccharides 996:1011 arg1 presence 965:972 the presence 961:972 the presence of fully-deacetylated oligosaccharides 961:1011 Neither hydrolysis nor transglycosylation by SpChiD was inhibited in the presence of fully-deacetylated oligosaccharides, suggesting that SpChiD could be exploited to generate oligosaccharides with defined DA and PA values. 26432054 3 40 with fractions 605:613 arg1 ranges 639:644 DA ranges 636:644 DA ranges 636:644 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 3 40 with fractions 605:613 arg1 DP 629:630 specific DP 620:630 specific DP 620:630 The crude oligosaccharide mixtures and purified fractions with specific DP and DA ranges were tested for their ability to induce an oxidative burst in rice cell suspension cultures. 26432054 1 41 theme oligosaccharides 172:187 arg1 activities 141:150 The biological activities 126:150 The biological activities of chitosan and its oligosaccharides 126:187 The biological activities of chitosan and its oligosaccharides are greatly influenced by properties such as the degree of polymerization (DP), degree of acetylation (DA) and pattern of acetylation (PA). 26432054 0 42 theme hypertransglycosylating 60:82 arg1 chitinase-D 84:94 the hypertransglycosylating chitinase-D 56:94 the hypertransglycosylating chitinase-D from Serratia proteamaculans 56:123 Production of bioactive chitosan oligosaccharides using the hypertransglycosylating chitinase-D from Serratia proteamaculans. 26432054 2 43 dep polymers 396:403 arg1 % 411:411 DA=35% 406:411 DA=35% 406:411 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 2 43 dep polymers 396:403 arg1 % 418:418 61% 416:418 61% 416:418 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 26432054 2 44 theme chitosan 356:363 arg1 oligosaccharides 365:380 structurally diverse chitosan oligosaccharides 335:380 structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) 335:419 Here, structurally diverse chitosan oligosaccharides from chitosan polymers (DA=35% or 61%) were generated using Serratia proteamaculans wild-type chitinase D (SpChiD) and the W114A mutant which lacks transglycosylase activity. 28328971 0 0 theme germinating 58:68 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 0 theme germinating 58:68 arg1 Oryza 41:45 Oryza 41:45 Oryza 41:45 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 1 1 theme life 124:127 arg1 cycle 129:133 the angiosperm life cycle 109:133 the angiosperm life cycle 109:133 Germination is a key event in the angiosperm life cycle. 28328971 7 2 theme glycosylation-mediated 1262:1283 arg1 mechanism 1325:1333 a key mechanism 1319:1333 a key mechanism regulating rice embryo germination 1319:1368 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 2 theme glycosylation-mediated 1262:1283 arg1 signaling 1302:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 3 theme several 1154:1160 arg1 proteins 1189:1196 several Brassinosteroids signaling proteins 1154:1196 several Brassinosteroids signaling proteins 1154:1196 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 3 theme several 1154:1160 arg1 XIAO 1209:1212 XIAO 1209:1212 XIAO 1209:1212 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 3 theme several 1154:1160 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 4 4 gly glycosites 580:589 arg2 glycosites 580:589 242 glycosites 576:589 242 glycosites from 191 unique proteins 576:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 2 5 theme embryo 312:317 arg1 proteome 284:291 the proteome 280:291 the proteome of the germinating embryo 280:317 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 8 6 theme rice 1505:1508 arg1 germination 1515:1525 rice seed germination 1505:1525 rice seed germination 1505:1525 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 3 7 gly glycosites 355:364 arg2 glycosites 355:364 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 0 8 link N-linked 12:19 arg1 glycosites 21:30 the N-linked glycosites 8:30 the N-linked glycosites of rice (Oryza sativa L.) germinating embryos 8:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 8 link N-linked 12:19 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 3 9 theme high 517:520 arg1 identification 549:562 high accuracy mass spectrometry identification 517:562 high accuracy mass spectrometry identification 517:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 2 10 gly N-glycosylation 136:150 arg1 proteins 155:162 proteins 155:162 proteins 155:162 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 0 11 theme embryos 70:76 arg1 glycosites 21:30 the N-linked glycosites 8:30 the N-linked glycosites of rice (Oryza sativa L.) germinating embryos 8:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 11 theme embryos 70:76 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 5 12 theme motifs 650:655 arg1 Inspection 632:641 Inspection 632:641 Inspection of the motifs and sequence structures involved 632:688 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 3 13 theme accuracy 522:529 arg1 identification 549:562 high accuracy mass spectrometry identification 517:562 high accuracy mass spectrometry identification 517:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 3 14 theme chromatography 444:457 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 4 15 from total 567:571 arg1 proteins 607:614 191 unique proteins 596:614 191 unique proteins 596:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 6 16 theme hydrolase 877:885 arg1 activities 887:896 glycoside hydrolase activities 867:896 glycoside hydrolase activities 867:896 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 3 17 gly glycopeptides 467:479 arg2 glycopeptides 467:479 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 6 18 gly N-glycosylation 810:824 arg2 proteins 853:860 proteins 853:860 proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway 853:976 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 7 19 theme signaling 1179:1187 arg1 proteins 1189:1196 several Brassinosteroids signaling proteins 1154:1196 several Brassinosteroids signaling proteins 1154:1196 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 19 theme signaling 1179:1187 arg1 XIAO 1209:1212 XIAO 1209:1212 XIAO 1209:1212 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 19 theme signaling 1179:1187 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 6 20 theme glycoside 867:875 arg1 activities 887:896 glycoside hydrolase activities 867:896 glycoside hydrolase activities 867:896 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 1 21 from event 100:104 arg1 cycle 129:133 the angiosperm life cycle 109:133 the angiosperm life cycle 109:133 Germination is a key event in the angiosperm life cycle. 28328971 5 22 dep motifs 650:655 arg1 the 646:648 the 646:648 the 646:648 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 7 23 theme interaction 1109:1119 arg1 analysis 1121:1128 protein-protein interaction analysis 1093:1128 protein-protein interaction analysis 1093:1128 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 24 theme BR-responsive 1224:1236 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 5 25 theme them 779:782 arg1 % 774:774 82.3% 770:774 82.3% of them 770:782 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 5 25 theme them 779:782 arg1 them 779:782 them 779:782 them 779:782 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 7 26 theme protein-protein 1093:1107 arg1 analysis 1121:1128 protein-protein interaction analysis 1093:1128 protein-protein interaction analysis 1093:1128 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 3 27 theme rice 377:380 arg1 embryos 382:388 rice embryos 377:388 rice embryos 377:388 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 3 28 theme first 340:344 arg1 mapping 366:372 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 3 29 theme spectrometry 536:547 arg1 identification 549:562 high accuracy mass spectrometry identification 517:562 high accuracy mass spectrometry identification 517:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 6 30 with proteins 853:860 arg1 activities 887:896 glycoside hydrolase activities 867:896 glycoside hydrolase activities 867:896 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 5 31 gly glycosites 713:722 arg2 glycosites 713:722 all the glycosites 705:722 all the glycosites 705:722 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 8 32 from regulation 1491:1500 arg1 germination 1515:1525 rice seed germination 1505:1525 rice seed germination 1505:1525 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 3 33 theme embryos 382:388 arg1 mapping 366:372 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 6 34 theme metabolism 959:968 arg1 pathway 970:976 the starch and sucrose metabolism pathway 936:976 the starch and sucrose metabolism pathway 936:976 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 8 35 theme seed 1510:1513 arg1 germination 1515:1525 rice seed germination 1505:1525 rice seed germination 1505:1525 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 8 36 theme novel 1464:1468 arg1 insight 1470:1476 novel insight 1464:1476 novel insight into the PTM regulation in rice seed germination 1464:1525 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 3 37 theme mass 531:534 arg1 identification 549:562 high accuracy mass spectrometry identification 517:562 high accuracy mass spectrometry identification 517:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 3 38 link N-linked 346:353 arg1 mapping 366:372 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 5 39 theme coil 794:797 arg1 structure 799:807 a coil structure 792:807 a coil structure 792:807 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 0 40 theme N-linked 12:19 arg1 glycosites 21:30 the N-linked glycosites 8:30 the N-linked glycosites of rice (Oryza sativa L.) germinating embryos 8:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 40 theme N-linked 12:19 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 6 41 theme sucrose 951:957 arg1 pathway 970:976 the starch and sucrose metabolism pathway 936:976 the starch and sucrose metabolism pathway 936:976 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 8 42 theme protein 1420:1426 arg1 glycosylation 1428:1440 protein glycosylation 1420:1440 protein glycosylation 1420:1440 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 2 43 theme proteins 155:162 arg1 regulator 267:275 an important regulator 254:275 an important regulator of the proteome of the germinating embryo 254:317 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 2 43 theme proteins 155:162 arg1 N-glycosylation 136:150 N-glycosylation 136:150 N-glycosylation of proteins 136:162 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 2 44 theme important 257:265 arg1 regulator 267:275 an important regulator 254:275 an important regulator of the proteome of the germinating embryo 254:317 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 2 44 theme important 257:265 arg1 N-glycosylation 136:150 N-glycosylation 136:150 N-glycosylation of proteins 136:162 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 3 45 theme hydrophilic 420:430 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 0 46 theme rice 35:38 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 46 theme rice 35:38 arg1 Oryza 41:45 Oryza 41:45 Oryza 41:45 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 3 47 theme HILIC 460:464 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 7 48 theme other 1218:1222 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 0 49 dep Oryza 41:45 arg1 L. 54:55 Oryza sativa L. 41:55 Oryza sativa L. 41:55 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 4 50 from proteins 607:614 arg1 total 567:571 A total 565:571 A total of 242 glycosites from 191 unique proteins 565:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 4 50 from proteins 607:614 arg1 glycosites 580:589 242 glycosites 576:589 242 glycosites from 191 unique proteins 576:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 6 51 theme starch 940:945 arg1 pathway 970:976 the starch and sucrose metabolism pathway 936:976 the starch and sucrose metabolism pathway 936:976 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 2 52 theme post-translational 190:207 arg1 modifications 209:221 the most common post-translational modifications 174:221 the most common post-translational modifications 174:221 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 3 53 theme N-linked 346:353 arg1 mapping 366:372 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 7 54 theme rice 1346:1349 arg1 germination 1358:1368 rice embryo germination 1346:1368 rice embryo germination 1346:1368 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 2 55 theme common 183:188 arg1 modifications 209:221 the most common post-translational modifications 174:221 the most common post-translational modifications 174:221 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 6 56 gly N-glycosylation 995:1009 arg1 germination 1033:1043 embryo germination 1026:1043 embryo germination 1026:1043 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 2 57 theme germinating 300:310 arg1 embryo 312:317 the germinating embryo 296:317 the germinating embryo 296:317 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 3 58 theme glycosites 355:364 arg1 mapping 366:372 the first N-linked glycosites mapping 336:372 the first N-linked glycosites mapping of rice embryos during germination 336:407 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 5 59 theme sequence 661:668 arg1 structures 670:679 sequence structures 661:679 sequence structures 661:679 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 0 60 gly glycosites 21:30 arg2 glycosites 21:30 the N-linked glycosites 8:30 the N-linked glycosites of rice (Oryza sativa L.) germinating embryos 8:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 60 gly glycosites 21:30 arg2 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 60 gly glycosites 21:30 arg1 embryos 70:76 rice (Oryza sativa L.) germinating embryos 35:76 rice (Oryza sativa L.) germinating embryos 35:76 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 0 60 gly glycosites 21:30 arg1 Oryza 41:45 Oryza 41:45 Oryza 41:45 Mapping the N-linked glycosites of rice (Oryza sativa L.) germinating embryos. 28328971 3 61 theme glycopeptides 467:479 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 1 62 theme key 96:98 arg1 event 100:104 a key event 94:104 a key event in the angiosperm life cycle 94:133 Germination is a key event in the angiosperm life cycle. 28328971 1 62 theme key 96:98 arg1 Germination 79:89 Germination 79:89 Germination 79:89 Germination is a key event in the angiosperm life cycle. 28328971 7 63 theme key 1321:1323 arg1 mechanism 1325:1333 a key mechanism 1319:1333 a key mechanism regulating rice embryo germination 1319:1368 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 63 theme key 1321:1323 arg1 signaling 1302:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 5 64 theme structures 670:679 arg1 Inspection 632:641 Inspection 632:641 Inspection of the motifs and sequence structures involved 632:688 Inspection of the motifs and sequence structures involved suggested that all the glycosites were concentrated within [NxS/T] motif, while 82.3% of them were in a coil structure. 28328971 8 65 theme glycosylation 1428:1440 arg1 knowledge 1407:1415 our knowledge 1403:1415 our knowledge of protein glycosylation in rice 1403:1448 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 3 66 theme interaction 432:442 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 6 67 theme embryo 1026:1031 arg1 germination 1033:1043 embryo germination 1026:1043 embryo germination 1026:1043 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 8 68 theme PTM 1487:1489 arg1 regulation 1491:1500 the PTM regulation 1483:1500 the PTM regulation in rice seed germination 1483:1525 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 2 69 theme proteome 284:291 arg1 regulator 267:275 an important regulator 254:275 an important regulator of the proteome of the germinating embryo 254:317 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 2 69 theme proteome 284:291 arg1 N-glycosylation 136:150 N-glycosylation 136:150 N-glycosylation of proteins 136:162 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 8 70 from knowledge 1407:1415 arg1 rice 1445:1448 rice 1445:1448 rice 1445:1448 In summary, this study expanded our knowledge of protein glycosylation in rice, and provided novel insight into the PTM regulation in rice seed germination. 28328971 4 71 theme glycosites 580:589 arg1 total 567:571 A total 565:571 A total of 242 glycosites from 191 unique proteins 565:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 7 72 theme Brassinosteroids 1285:1300 arg1 mechanism 1325:1333 a key mechanism 1319:1333 a key mechanism regulating rice embryo germination 1319:1368 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 72 theme Brassinosteroids 1285:1300 arg1 signaling 1302:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 glycosylation-mediated Brassinosteroids signaling 1262:1310 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 73 with network 1141:1147 arg1 proteins 1189:1196 several Brassinosteroids signaling proteins 1154:1196 several Brassinosteroids signaling proteins 1154:1196 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 73 with network 1141:1147 arg1 XIAO 1209:1212 XIAO 1209:1212 XIAO 1209:1212 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 73 with network 1141:1147 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 6 74 theme carbohydrate 1059:1070 arg1 metabolism 1072:1081 carbohydrate metabolism 1059:1081 carbohydrate metabolism 1059:1081 N-glycosylation preferentially occurred on proteins with glycoside hydrolase activities, which were significantly enriched in the starch and sucrose metabolism pathway, suggesting that N-glycosylation is involved in embryo germination by regulating carbohydrate metabolism. 28328971 3 75 theme enrichment 481:490 arg1 strategy 492:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy 418:499 a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification 418:562 Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. 28328971 4 76 theme unique 600:605 arg1 proteins 607:614 191 unique proteins 596:614 191 unique proteins 596:614 A total of 242 glycosites from 191 unique proteins was discovered. 28328971 7 77 theme Brassinosteroids 1162:1177 arg1 proteins 1189:1196 several Brassinosteroids signaling proteins 1154:1196 several Brassinosteroids signaling proteins 1154:1196 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 77 theme Brassinosteroids 1162:1177 arg1 XIAO 1209:1212 XIAO 1209:1212 XIAO 1209:1212 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 77 theme Brassinosteroids 1162:1177 arg1 proteins 1238:1245 other BR-responsive proteins 1218:1245 other BR-responsive proteins 1218:1245 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 7 78 theme embryo 1351:1356 arg1 germination 1358:1368 rice embryo germination 1346:1368 rice embryo germination 1346:1368 Notably, protein-protein interaction analysis revealed a network with several Brassinosteroids signaling proteins, including XIAO and other BR-responsive proteins, implying that glycosylation-mediated Brassinosteroids signaling may be a key mechanism regulating rice embryo germination. 28328971 2 79 theme modifications 209:221 arg1 one 167:169 one 167:169 one 167:169 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 2 79 theme modifications 209:221 arg1 modifications 209:221 the most common post-translational modifications 174:221 the most common post-translational modifications 174:221 N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. 28328971 1 80 theme angiosperm 113:122 arg1 cycle 129:133 the angiosperm life cycle 109:133 the angiosperm life cycle 109:133 Germination is a key event in the angiosperm life cycle. 28942028 1 0 theme cell 297:300 arg1 walls 302:306 the cell walls 293:306 the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550 293:363 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 0 1 theme Arthrobacter 163:174 arg1 crystallopoietes 176:191 Arthrobacter crystallopoietes 163:191 Arthrobacter crystallopoietes 163:191 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 5 2 theme A. crystallopoietes 935:953 arg1 strain 884:889 the type strain 875:889 the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer 875:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 2 3 theme Arthrobacter 434:445 arg1 representatives 405:419 representatives 405:419 representatives of the genus Arthrobacter for the first time 405:464 The teichulosonic acid was revealed in representatives of the genus Arthrobacter for the first time. 28942028 5 4 theme glycosyl 1022:1029 arg1 polymer 1043:1049 a glycosyl 1-phosphate polymer 1020:1049 a glycosyl 1-phosphate polymer 1020:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 1 5 theme -β-D-GlcpNAc- 246:258 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 5 6 theme 1-phosphate 1031:1041 arg1 polymer 1043:1049 a glycosyl 1-phosphate polymer 1020:1049 a glycosyl 1-phosphate polymer 1020:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 0 7 theme Arthrobacter 82:93 arg1 walls 73:77 the cell walls 64:77 the cell walls of Arthrobacter sp., strains 64:106 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 5 8 theme teichoic 987:994 arg1 acid 996:999 a teichoic acid 985:999 a teichoic acid 985:999 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 5 9 theme cell 848:851 arg1 composition 858:868 the cell wall composition 844:868 the cell wall composition 844:868 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 1 10 theme 1→8 260:262 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 11 theme Arthrobacter 315:326 arg1 strains 328:334 two Arthrobacter strains 311:334 two Arthrobacter strains 311:334 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 11 theme Arthrobacter 315:326 arg1 Ac-2549 341:347 VKM Ac-2549 337:347 VKM Ac-2549 337:347 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 11 theme Arthrobacter 315:326 arg1 Ac-2550 357:363 VKM Ac-2550 353:363 VKM Ac-2550 353:363 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 5 12 theme wall 853:856 arg1 composition 858:868 the cell wall composition 844:868 the cell wall composition 844:868 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 1 13 theme strains 328:334 arg1 walls 302:306 the cell walls 293:306 the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550 293:363 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 2 14 theme genus 428:432 arg1 Arthrobacter 434:445 the genus Arthrobacter 424:445 the genus Arthrobacter for the first time 424:464 The teichulosonic acid was revealed in representatives of the genus Arthrobacter for the first time. 28942028 0 15 dep Arthrobacter 82:93 arg1 sp. 95:97 Arthrobacter sp., strains 82:106 sp. 95:97 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 15 dep Arthrobacter 82:93 arg1 strains 100:106 strains 100:106 strains 100:106 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 16 theme teichulosonic 6:18 arg1 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 2 17 theme first 455:459 arg1 time 461:464 the first time 451:464 the first time 451:464 The teichulosonic acid was revealed in representatives of the genus Arthrobacter for the first time. 28942028 0 18 theme Novel 0:4 arg1 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 3 19 theme -β-D-GlcpNAc- 655:667 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 19 theme -β-D-GlcpNAc- 655:667 arg1 1-P- 684:687 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 684:687 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 1 20 theme -α-Kdn- 264:270 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 21 with acid 214:217 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 5 22 theme related 919:925 arg1 A. crystallopoietes 935:953 phylogenetically closely related species A. crystallopoietes 894:953 phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer 894:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 3 23 theme above 504:508 arg1 strains 510:516 the above strains 500:516 the above strains 500:516 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 1 24 theme Novel 194:198 arg1 acid 214:217 Novel teichulosonic acid 194:217 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 194:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 5 25 contain contain 977:983 arg1 A. crystallopoietes 935:953 phylogenetically closely related species A. crystallopoietes 894:953 phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer 894:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 5 25 contain contain 977:983 arg2 acid 996:999 a teichoic acid 985:999 a teichoic acid 985:999 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 1 26 theme VKM 337:339 arg1 strains 328:334 two Arthrobacter strains 311:334 two Arthrobacter strains 311:334 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 26 theme VKM 337:339 arg1 Ac-2549 341:347 VKM Ac-2549 337:347 VKM Ac-2549 337:347 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 27 located found 284:288 arg1 walls 302:306 the cell walls 293:306 the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550 293:363 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 27 located found 284:288 arg2 acid 214:217 Novel teichulosonic acid 194:217 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 194:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 28 theme teichulosonic 200:212 arg1 acid 214:217 Novel teichulosonic acid 194:217 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 194:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 0 29 theme 1-phosphate 38:48 arg1 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 30 theme VKM 124:126 arg1 Ac-2550 128:134 VKM Ac-2550 124:134 VKM Ac-2550 124:134 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 31 from walls 73:77 arg1 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 acid 20:23 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 31 from walls 73:77 arg1 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 0 32 theme glycosyl 29:36 arg1 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers 0:57 polymers 50:57 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 3 33 theme -α-D-Galp- 673:682 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 33 theme -α-D-Galp- 673:682 arg1 1-P- 684:687 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 684:687 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 1 34 theme VKM 353:355 arg1 strains 328:334 two Arthrobacter strains 311:334 two Arthrobacter strains 311:334 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 1 34 theme VKM 353:355 arg1 Ac-2550 357:363 VKM Ac-2550 353:363 VKM Ac-2550 353:363 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 4 35 theme polymers 718:725 arg1 structures 694:703 The structures 690:703 The structures of all three polymers 690:725 The structures of all three polymers were established by using chemical, NMR spectroscopic and ESI-MS methods. 28942028 4 36 dep chemical 753:760 arg1 methods 792:798 methods 792:798 methods 792:798 The structures of all three polymers were established by using chemical, NMR spectroscopic and ESI-MS methods. 28942028 3 37 theme 1→3 669:671 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 37 theme 1→3 669:671 arg1 1-P- 684:687 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 684:687 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 5 38 theme species 927:933 arg1 A. crystallopoietes 935:953 phylogenetically closely related species A. crystallopoietes 894:953 phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer 894:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 3 39 theme monoglycosyl 528:539 arg1 1-phosphate 541:551 monoglycosyl 1-phosphate 528:551 monoglycosyl 1-phosphate 528:551 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 39 theme monoglycosyl 528:539 arg1 poly 523:526 poly 523:526 poly(monoglycosyl 1-phosphate) 523:552 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 40 theme i.e. 619:622 arg1 1-P- 642:645 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 642:645 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 40 theme i.e. 619:622 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 41 theme unknown 599:605 arg1 structures 607:616 hitherto unknown structures 590:616 hitherto unknown structures 590:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 42 theme diglycosyl 563:572 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 42 theme diglycosyl 563:572 arg1 1-phosphate 574:584 diglycosyl 1-phosphate 563:584 diglycosyl 1-phosphate 563:584 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 1 43 theme repeating 228:236 arg1 unit 238:241 the repeating unit 224:241 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 0 44 theme cell 68:71 arg1 walls 73:77 the cell walls 64:77 the cell walls of Arthrobacter sp., strains 64:106 Novel teichulosonic acid and glycosyl 1-phosphate polymers from the cell walls of Arthrobacter sp., strains VKM Ac-2549 and VKM Ac-2550, phylogenetically close to Arthrobacter crystallopoietes. 28942028 3 45 theme structures 607:616 arg1 1-phosphate 541:551 monoglycosyl 1-phosphate 528:551 monoglycosyl 1-phosphate 528:551 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 45 theme structures 607:616 arg1 1-P- 642:645 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 642:645 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 45 theme structures 607:616 arg1 1-phosphate 574:584 diglycosyl 1-phosphate 563:584 diglycosyl 1-phosphate 563:584 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 45 theme structures 607:616 arg1 poly 523:526 poly 523:526 poly(monoglycosyl 1-phosphate) 523:552 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 45 theme structures 607:616 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 45 theme structures 607:616 arg1 1-P- 684:687 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 684:687 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 5 46 theme type 879:882 arg1 strain 884:889 the type strain 875:889 the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer 875:1049 The strains studied in this work differ in the cell wall composition from the type strain of phylogenetically closely related species A. crystallopoietes which was reported to contain a teichoic acid and supposedly had a glycosyl 1-phosphate polymer. 28942028 2 47 theme teichulosonic 370:382 arg1 acid 384:387 The teichulosonic acid 366:387 The teichulosonic acid 366:387 The teichulosonic acid was revealed in representatives of the genus Arthrobacter for the first time. 28942028 1 48 theme unit 238:241 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 28942028 3 49 theme -α-D-GalpNAc- 628:640 arg1 1-P- 642:645 i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P- 619:687 1-P- 642:645 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 49 theme -α-D-GalpNAc- 628:640 arg1 poly 558:561 poly 558:561 poly(diglycosyl 1-phosphate) of hitherto unknown structures 558:616 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 3 50 theme other 471:475 arg1 polymers 477:484 Two other polymers 467:484 Two other polymers identified in the above strains 467:516 Two other polymers identified in the above strains were poly(monoglycosyl 1-phosphate) and poly(diglycosyl 1-phosphate) of hitherto unknown structures, i.e., -6)-α-D-GalpNAc-(1-P-, and -6)-β-D-GlcpNAc-(1→3)-α-D-Galp-(1-P-. 28942028 1 51 theme →6 243:244 arg1 2→ 272:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ 224:273 Novel teichulosonic acid with the repeating unit →6)-β-D-GlcpNAc-(1→8)-α-Kdn-(2→ has been found in the cell walls of two Arthrobacter strains, VKM Ac-2549 and VKM Ac-2550. 26840264 2 0 theme IgGs 352:355 arg1 levels 326:331 increased plasmatic levels 306:331 increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained 306:447 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 1 1 theme anti-inflammatory 86:102 arg1 activities 104:113 The pro- or anti-inflammatory activities 74:113 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) 74:141 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 6 2 theme whole 1304:1308 arg1 range 1314:1318 the whole age range 1300:1318 the whole age range 1300:1318 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 6 3 theme B4GALTs 1220:1226 arg1 marker 1246:1251 a new marker 1240:1251 a new marker of aging 1240:1260 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 6 3 theme B4GALTs 1220:1226 arg1 activity 1228:1235 1) plasmatic B4GALTs activity 1207:1235 1) plasmatic B4GALTs activity 1207:1235 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 9 4 dep lived 1616:1620 arg1 short 1610:1614 short 1610:1614 short 1610:1614 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 3 5 theme cell 559:562 arg1 membranes 564:572 internal cell membranes 550:572 internal cell membranes 550:572 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 5 6 theme plasmatic 950:958 arg1 s 985:985 s 985:985 s 985:985 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 6 theme plasmatic 950:958 arg1 B4GALTs 989:995 B4GALTs 989:995 B4GALTs 989:995 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 6 theme plasmatic 950:958 arg1 galactosyltransferase 963:983 plasmatic β4 galactosyltransferase 950:983 plasmatic β4 galactosyltransferase(s) (B4GALTs) 950:996 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 6 7 theme linear 1273:1278 arg1 increase 1280:1287 a linear increase 1271:1287 a linear increase throughout the whole age range 1271:1318 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 10 8 theme plasmatic 1761:1769 arg1 glycosyltransferases 1771:1790 the two plasmatic glycosyltransferases 1753:1790 the two plasmatic glycosyltransferases 1753:1790 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 1 9 theme glycan 182:187 arg1 structure 165:173 the structure 161:173 the structure of the glycan N-linked to Asn297 of their heavy chain 161:227 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 5 10 dep test 1076:1079 arg1 marker 1104:1109 a glycosylation-based marker 1082:1109 a glycosylation-based marker of aging 1082:1118 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 11 theme plasma 1129:1134 arg1 level 1136:1140 the plasma level 1125:1140 the plasma level of inflammatory and liver damage markers 1125:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 3 12 theme extracellular 579:591 arg1 glycosylation 593:605 the extracellular glycosylation 575:605 the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases 575:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 3 13 gly glycosylation 593:605 arg1 glycoproteins 622:634 circulating glycoproteins 610:634 circulating glycoproteins mediated by plasmatic glycosyltransferases 610:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 3 14 gly glycoproteins 622:634 arg1 glycoproteins 622:634 circulating glycoproteins 610:634 circulating glycoproteins mediated by plasmatic glycosyltransferases 610:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 0 15 theme plasma 24:29 arg1 markers 56:62 novel plasma glycosylation-associated markers 18:62 novel plasma glycosylation-associated markers of aging 18:71 Identification of novel plasma glycosylation-associated markers of aging. 26840264 9 16 with association 1556:1566 arg1 phase 1579:1583 acute phase proteins 1573:1592 acute phase proteins 1573:1592 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 5 17 theme aging 1114:1118 arg1 marker 1104:1109 a glycosylation-based marker 1082:1109 a glycosylation-based marker of aging 1082:1118 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 3 18 theme glycoproteins 622:634 arg1 glycosylation 593:605 the extracellular glycosylation 575:605 the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases 575:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 10 19 with consistent 1838:1847 arg1 shift 1856:1860 a shift 1854:1860 a shift toward a pro-inflammatory glycotype 1854:1896 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 4 20 theme IgG 803:805 arg1 glycosylation 807:819 IgG glycosylation 803:819 IgG glycosylation 803:819 In this study we have investigated the relationship between plasmatic glycosyltransferases, IgG glycosylation and inflammatory and aging markers. 26840264 2 21 theme low 249:251 arg1 inflammaging 273:284 inflammaging 273:284 inflammaging 273:284 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 2 21 theme low 249:251 arg1 inflammation 259:270 The age-associated low grade inflammation 230:270 The age-associated low grade inflammation (inflammaging) 230:285 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 6 22 theme new 1242:1244 arg1 marker 1246:1251 a new marker 1240:1251 a new marker of aging 1240:1260 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 6 22 theme new 1242:1244 arg1 activity 1228:1235 1) plasmatic B4GALTs activity 1207:1235 1) plasmatic B4GALTs activity 1207:1235 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 10 23 theme age-associated 1815:1828 arg1 changes 1830:1836 progressive age-associated changes 1803:1836 progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype 1803:1896 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 8 24 theme damage 1509:1514 arg1 markers 1492:1498 markers 1492:1498 markers of liver damage 1492:1514 3) Neither plasmatic glycosyltransferase correlated with markers of liver damage. 26840264 9 25 dep phase 1579:1583 arg1 proteins 1585:1592 proteins 1585:1592 acute phase proteins 1573:1592 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 10 26 theme consistent 1838:1847 arg1 changes 1830:1836 progressive age-associated changes 1803:1836 progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype 1803:1896 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 1 27 link N-linked 189:196 arg1 glycan 182:187 the glycan 178:187 the glycan N-linked to Asn297 of their heavy chain 178:227 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 4 28 theme inflammatory 825:836 arg1 markers 848:854 inflammatory and aging markers 825:854 markers 848:854 In this study we have investigated the relationship between plasmatic glycosyltransferases, IgG glycosylation and inflammatory and aging markers. 26840264 10 29 gly glycosylation 1694:1706 arg1 IgGs 1711:1714 IgGs 1711:1714 IgGs 1711:1714 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 5 30 theme damage 1168:1173 arg1 markers 1175:1181 inflammatory and liver damage markers 1145:1181 markers 1175:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 9 31 theme acute 1573:1577 arg1 phase 1579:1583 acute phase proteins 1573:1592 acute phase proteins 1573:1592 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 5 32 theme ST6GAL1 1028:1034 arg1 test 1076:1079 the GlycoAge test 1063:1079 the GlycoAge test (a glycosylation-based marker of aging) 1063:1119 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 32 theme ST6GAL1 1028:1034 arg1 level 1136:1140 the plasma level 1125:1140 the plasma level of inflammatory and liver damage markers 1125:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 32 theme ST6GAL1 1028:1034 arg1 activity 938:945 the activity 934:945 the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1 934:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 32 theme ST6GAL1 1028:1034 arg1 glycosylation 1041:1053 the glycosylation 1037:1053 the glycosylation of IgG 1037:1060 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 2 33 theme age-associated 234:247 arg1 inflammaging 273:284 inflammaging 273:284 inflammaging 273:284 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 2 33 theme age-associated 234:247 arg1 inflammation 259:270 The age-associated low grade inflammation 230:270 The age-associated low grade inflammation (inflammaging) 230:285 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 0 34 theme novel 18:22 arg1 markers 56:62 novel plasma glycosylation-associated markers 18:62 novel plasma glycosylation-associated markers of aging 18:71 Identification of novel plasma glycosylation-associated markers of aging. 26840264 1 35 theme immunoglobulins 118:132 arg1 IgGs 137:140 IgGs 137:140 IgGs 137:140 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 1 35 theme immunoglobulins 118:132 arg1 G 134:134 immunoglobulins G 118:134 immunoglobulins G (IgGs) 118:141 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 0 36 theme glycosylation-associated 31:54 arg1 markers 56:62 novel plasma glycosylation-associated markers 18:62 novel plasma glycosylation-associated markers of aging 18:71 Identification of novel plasma glycosylation-associated markers of aging. 26840264 8 37 dep correlated 1476:1485 arg1 3 1435:1435 3 1435:1435 3 1435:1435 3) Neither plasmatic glycosyltransferase correlated with markers of liver damage. 26840264 5 38 theme glycosylation-based 1084:1102 arg1 marker 1104:1109 a glycosylation-based marker 1082:1109 a glycosylation-based marker of aging 1082:1118 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 2 39 theme agalactosylated 336:350 arg1 IgGs 352:355 agalactosylated IgGs 336:355 agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained 336:447 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 9 40 theme parents 1622:1628 arg1 offspring 1597:1605 offspring 1597:1605 offspring of short lived parents 1597:1628 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 7 41 dep high 1346:1349 arg1 2 1321:1321 2 1321:1321 2 1321:1321 2) plasmatic ST6GAL1 was high only in children and in people above 80, showing a quadratic relationship with age. 26840264 5 42 theme GlycoAge 1067:1074 arg1 test 1076:1079 the GlycoAge test 1063:1079 the GlycoAge test (a glycosylation-based marker of aging) 1063:1119 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 10 43 theme pro-inflammatory 1871:1886 arg1 glycotype 1888:1896 a pro-inflammatory glycotype 1869:1896 a pro-inflammatory glycotype 1869:1896 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 1 44 theme chain 223:227 arg1 Asn297 201:206 Asn297 201:206 Asn297 of their heavy chain 201:227 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 0 45 theme aging 67:71 arg1 markers 56:62 novel plasma glycosylation-associated markers 18:62 novel plasma glycosylation-associated markers of aging 18:71 Identification of novel plasma glycosylation-associated markers of aging. 26840264 6 46 theme age 1310:1312 arg1 range 1314:1318 the whole age range 1300:1318 the whole age range 1300:1318 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 10 47 theme glycosyltransferases 1771:1790 arg1 level 1744:1748 the level 1740:1748 the level of the two plasmatic glycosyltransferases 1740:1790 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 3 48 theme internal 550:557 arg1 membranes 564:572 internal cell membranes 550:572 internal cell membranes 550:572 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 6 49 theme plasmatic 1210:1218 arg1 marker 1246:1251 a new marker 1240:1251 a new marker of aging 1240:1260 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 6 49 theme plasmatic 1210:1218 arg1 activity 1228:1235 1) plasmatic B4GALTs activity 1207:1235 1) plasmatic B4GALTs activity 1207:1235 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 2 50 theme increased 306:314 arg1 levels 326:331 increased plasmatic levels 306:331 increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained 306:447 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 5 51 theme β4 960:961 arg1 s 985:985 s 985:985 s 985:985 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 51 theme β4 960:961 arg1 B4GALTs 989:995 B4GALTs 989:995 B4GALTs 989:995 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 51 theme β4 960:961 arg1 galactosyltransferase 963:983 plasmatic β4 galactosyltransferase 950:983 plasmatic β4 galactosyltransferase(s) (B4GALTs) 950:996 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 1 52 theme N-linked 189:196 arg1 glycan 182:187 the glycan 178:187 the glycan N-linked to Asn297 of their heavy chain 178:227 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 8 53 theme plasmatic 1446:1454 arg1 glycosyltransferase 1456:1474 Neither plasmatic glycosyltransferase 1438:1474 Neither plasmatic glycosyltransferase 1438:1474 3) Neither plasmatic glycosyltransferase correlated with markers of liver damage. 26840264 7 54 theme quadratic 1402:1410 arg1 relationship 1412:1423 a quadratic relationship 1400:1423 a quadratic relationship with age 1400:1432 2) plasmatic ST6GAL1 was high only in children and in people above 80, showing a quadratic relationship with age. 26840264 10 55 theme progressive 1803:1813 arg1 changes 1830:1836 progressive age-associated changes 1803:1836 progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype 1803:1896 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 5 56 theme galactosyltransferase 963:983 arg1 test 1076:1079 the GlycoAge test 1063:1079 the GlycoAge test (a glycosylation-based marker of aging) 1063:1119 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 56 theme galactosyltransferase 963:983 arg1 level 1136:1140 the plasma level 1125:1140 the plasma level of inflammatory and liver damage markers 1125:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 56 theme galactosyltransferase 963:983 arg1 activity 938:945 the activity 934:945 the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1 934:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 56 theme galactosyltransferase 963:983 arg1 glycosylation 1041:1053 the glycosylation 1037:1053 the glycosylation of IgG 1037:1060 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 6 57 dep activity 1228:1235 arg1 1 1207:1207 1 1207:1207 1 1207:1207 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 5 58 theme inflammatory 1145:1156 arg1 markers 1175:1181 inflammatory and liver damage markers 1145:1181 markers 1175:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 3 59 theme circulating 610:620 arg1 glycoproteins 622:634 circulating glycoproteins 610:634 circulating glycoproteins mediated by plasmatic glycosyltransferases 610:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 1 60 theme G 134:134 arg1 activities 104:113 The pro- or anti-inflammatory activities 74:113 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) 74:141 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 6 61 theme aging 1256:1260 arg1 marker 1246:1251 a new marker 1240:1251 a new marker of aging 1240:1260 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 6 61 theme aging 1256:1260 arg1 activity 1228:1235 1) plasmatic B4GALTs activity 1207:1235 1) plasmatic B4GALTs activity 1207:1235 Our results show that: 1) plasmatic B4GALTs activity is a new marker of aging, showing a linear increase throughout the whole age range. 26840264 4 62 theme plasmatic 771:779 arg1 glycosyltransferases 781:800 plasmatic glycosyltransferases 771:800 plasmatic glycosyltransferases 771:800 In this study we have investigated the relationship between plasmatic glycosyltransferases, IgG glycosylation and inflammatory and aging markers. 26840264 2 63 theme grade 253:257 arg1 inflammaging 273:284 inflammaging 273:284 inflammaging 273:284 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 2 63 theme grade 253:257 arg1 inflammation 259:270 The age-associated low grade inflammation 230:270 The age-associated low grade inflammation (inflammaging) 230:285 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 8 64 theme liver 1503:1507 arg1 damage 1509:1514 liver damage 1503:1514 liver damage 1503:1514 3) Neither plasmatic glycosyltransferase correlated with markers of liver damage. 26840264 3 65 theme plasmatic 648:656 arg1 glycosyltransferases 658:677 plasmatic glycosyltransferases 648:677 plasmatic glycosyltransferases 648:677 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 9 66 theme plasmatic 1520:1528 arg1 ST6GAL1 1530:1536 4) plasmatic ST6GAL1 1517:1536 4) plasmatic ST6GAL1 1517:1536 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 5 67 gly glycosylation 1041:1053 arg1 IgG 1058:1060 IgG 1058:1060 IgG 1058:1060 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 s 985:985 s 985:985 s 985:985 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 markers 1175:1181 inflammatory and liver damage markers 1145:1181 markers 1175:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 ST6GAL1 1028:1034 α2,6-sialyltransferase ST6GAL1 1005:1034 α2,6-sialyltransferase ST6GAL1 1005:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 cohorts 860:866 cohorts 860:866 cohorts of individuals ranging from infancy to centenarians 860:918 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 B4GALTs 989:995 B4GALTs 989:995 B4GALTs 989:995 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 67 gly glycosylation 1041:1053 arg1 galactosyltransferase 963:983 plasmatic β4 galactosyltransferase 950:983 plasmatic β4 galactosyltransferase(s) (B4GALTs) 950:996 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 10 68 theme IgGs 1711:1714 arg1 glycosylation 1694:1706 the glycosylation 1690:1706 the glycosylation of IgGs 1690:1714 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 26840264 9 69 theme positive 1547:1554 arg1 association 1556:1566 a positive association 1545:1566 a positive association with acute phase proteins 1545:1592 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 4 70 theme aging 842:846 arg1 markers 848:854 inflammatory and aging markers 825:854 markers 848:854 In this study we have investigated the relationship between plasmatic glycosyltransferases, IgG glycosylation and inflammatory and aging markers. 26840264 5 71 theme liver 1162:1166 arg1 markers 1175:1181 inflammatory and liver damage markers 1145:1181 markers 1175:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 1 72 theme pro- 78:81 arg1 activities 104:113 The pro- or anti-inflammatory activities 74:113 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) 74:141 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 5 73 theme markers 1175:1181 arg1 test 1076:1079 the GlycoAge test 1063:1079 the GlycoAge test (a glycosylation-based marker of aging) 1063:1119 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 73 theme markers 1175:1181 arg1 level 1136:1140 the plasma level 1125:1140 the plasma level of inflammatory and liver damage markers 1125:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 73 theme markers 1175:1181 arg1 activity 938:945 the activity 934:945 the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1 934:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 73 theme markers 1175:1181 arg1 glycosylation 1041:1053 the glycosylation 1037:1053 the glycosylation of IgG 1037:1060 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 74 theme α2,6-sialyltransferase 1005:1026 arg1 ST6GAL1 1028:1034 α2,6-sialyltransferase ST6GAL1 1005:1034 α2,6-sialyltransferase ST6GAL1 1005:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 7 75 with relationship 1412:1423 arg1 age 1430:1432 age 1430:1432 age 1430:1432 2) plasmatic ST6GAL1 was high only in children and in people above 80, showing a quadratic relationship with age. 26840264 7 76 theme plasmatic 1324:1332 arg1 ST6GAL1 1334:1340 plasmatic ST6GAL1 1324:1340 plasmatic ST6GAL1 1324:1340 2) plasmatic ST6GAL1 was high only in children and in people above 80, showing a quadratic relationship with age. 26840264 5 77 theme IgG 1058:1060 arg1 test 1076:1079 the GlycoAge test 1063:1079 the GlycoAge test (a glycosylation-based marker of aging) 1063:1119 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 77 theme IgG 1058:1060 arg1 level 1136:1140 the plasma level 1125:1140 the plasma level of inflammatory and liver damage markers 1125:1181 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 77 theme IgG 1058:1060 arg1 activity 938:945 the activity 934:945 the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1 934:1034 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 5 77 theme IgG 1058:1060 arg1 glycosylation 1041:1053 the glycosylation 1037:1053 the glycosylation of IgG 1037:1060 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 9 78 dep ST6GAL1 1530:1536 arg1 4 1517:1517 4 1517:1517 4 1517:1517 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 3 79 theme glycans 479:485 arg1 biosynthesis 463:474 the biosynthesis 459:474 the biosynthesis of glycans 459:485 Although the biosynthesis of glycans is in general mediated by glycosyltransferases associated with internal cell membranes, the extracellular glycosylation of circulating glycoproteins mediated by plasmatic glycosyltransferases has been recently demonstrated. 26840264 0 80 theme markers 56:62 arg1 Identification 0:13 Identification 0:13 Identification of novel plasma glycosylation-associated markers of aging. 0:72 Identification of novel plasma glycosylation-associated markers of aging. 26840264 5 81 theme individuals 871:881 arg1 cohorts 860:866 cohorts 860:866 cohorts of individuals ranging from infancy to centenarians 860:918 In cohorts of individuals ranging from infancy to centenarians we determined the activity of plasmatic β4 galactosyltransferase(s) (B4GALTs) and of α2,6-sialyltransferase ST6GAL1, the glycosylation of IgG, the GlycoAge test (a glycosylation-based marker of aging) and the plasma level of inflammatory and liver damage markers. 26840264 9 82 theme lived 1616:1620 arg1 parents 1622:1628 short lived parents 1610:1628 short lived parents 1610:1628 4) plasmatic ST6GAL1 showed a positive association with acute phase proteins in offspring of short lived parents, but not in centenarians or in their offspring. 26840264 2 83 theme plasmatic 316:324 arg1 levels 326:331 increased plasmatic levels 306:331 increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained 306:447 The age-associated low grade inflammation (inflammaging) is associated with increased plasmatic levels of agalactosylated IgGs terminating with N-acetylglucosamine (IgG-G0) whose biogenesis has not been fully explained. 26840264 1 84 theme heavy 217:221 arg1 chain 223:227 their heavy chain 211:227 their heavy chain 211:227 The pro- or anti-inflammatory activities of immunoglobulins G (IgGs) are controlled by the structure of the glycan N-linked to Asn297 of their heavy chain. 26840264 10 85 dep showed 1796:1801 arg1 5 1678:1678 5 1678:1678 5 1678:1678 5) Although the glycosylation of IgGs was not correlated with the level of the two plasmatic glycosyltransferases, it showed progressive age-associated changes consistent with a shift toward a pro-inflammatory glycotype. 28924016 0 0 theme Glycan-Directed 73:87 arg1 Antibodies 89:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 5 1 theme xyloglucan-directed 811:829 arg1 antibodies 831:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 Our results reveal the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies. 28924016 4 2 theme plant 669:673 arg1 antibodies 701:710 plant cell wall glycan-directed antibodies 669:710 plant cell wall glycan-directed antibodies 669:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 6 3 with glycans 1099:1105 arg1 precision 1136:1144 unprecedented molecular precision 1112:1144 unprecedented molecular precision 1112:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 6 4 theme wall 1010:1013 arg1 analyses 1015:1022 immunological cell wall analyses 991:1022 immunological cell wall analyses 991:1022 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 0 5 theme Wall 68:71 arg1 Antibodies 89:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 1 6 theme monoclonal 142:151 arg1 antibodies 153:162 more than 200 monoclonal antibodies 128:162 more than 200 monoclonal antibodies 128:162 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 1 7 theme different 240:248 arg1 laboratories 250:261 different laboratories worldwide 240:271 different laboratories worldwide 240:271 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 6 8 theme epitopes 892:899 arg1 knowledge 869:877 knowledge 869:877 knowledge of the exact epitopes recognized by individual antibodies 869:935 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 2 9 from differences 319:329 arg1 components 350:359 plant cell wall components 334:359 plant cell wall components 334:359 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 2 9 from differences 319:329 arg1 stages 415:420 developmental stages 401:420 developmental stages 401:420 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 2 9 from differences 319:329 arg1 types 390:394 organ and tissue types 373:394 organ and tissue types 373:394 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 2 9 from differences 319:329 arg1 mutants 364:370 mutants 364:370 mutants 364:370 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 6 10 theme immunological 991:1003 arg1 analyses 1015:1022 immunological cell wall analyses 991:1022 immunological cell wall analyses 991:1022 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 0 11 theme Antibodies 89:98 arg1 Mapping 46:52 Epitope Mapping 38:52 Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies 38:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 6 12 theme exact 886:890 arg1 epitopes 892:899 the exact epitopes 882:899 the exact epitopes recognized by individual antibodies 882:935 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 6 13 theme molecular 1126:1134 arg1 precision 1136:1144 unprecedented molecular precision 1112:1144 unprecedented molecular precision 1112:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 1 14 theme last 108:111 arg1 decades 119:125 the last three decades 104:125 the last three decades 104:125 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 6 15 theme cell 1089:1092 arg1 glycans 1099:1105 plant cell wall glycans 1083:1105 plant cell wall glycans with unprecedented molecular precision 1083:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 2 16 theme tissue 383:388 arg1 types 390:394 organ and tissue types 373:394 organ and tissue types 373:394 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 3 17 theme precise 467:473 arg1 epitope 483:489 the precise binding epitope 463:489 the precise binding epitope 463:489 Despite their importance and broad use, the precise binding epitope has been determined for only a few of these antibodies. 28924016 6 18 theme plant 1083:1087 arg1 glycans 1099:1105 plant cell wall glycans 1083:1105 plant cell wall glycans with unprecedented molecular precision 1083:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 0 19 theme Synthetic 2:10 arg1 Microarray 19:28 A Synthetic Glycan Microarray 0:28 A Synthetic Glycan Microarray 0:28 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 3 20 theme antibodies 535:544 arg1 few 522:524 few 522:524 few 522:524 Despite their importance and broad use, the precise binding epitope has been determined for only a few of these antibodies. 28924016 6 21 theme wall 1094:1097 arg1 glycans 1099:1105 plant cell wall glycans 1083:1105 plant cell wall glycans with unprecedented molecular precision 1083:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 2 22 theme organ 373:377 arg1 types 390:394 organ and tissue types 373:394 organ and tissue types 373:394 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 5 23 theme xylan- 799:804 arg1 antibodies 831:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 Our results reveal the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies. 28924016 2 24 theme developmental 401:413 arg1 stages 415:420 developmental stages 401:420 developmental stages 401:420 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 0 25 theme Glycan 12:17 arg1 Microarray 19:28 A Synthetic Glycan Microarray 0:28 A Synthetic Glycan Microarray 0:28 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 2 26 theme wall 345:348 arg1 components 350:359 plant cell wall components 334:359 plant cell wall components 334:359 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 1 27 theme most 189:192 arg1 classes 194:200 most classes 189:200 most classes of plant cell wall polysaccharides 189:235 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 0 28 theme Epitope 38:44 arg1 Mapping 46:52 Epitope Mapping 38:52 Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies 38:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 3 29 theme binding 475:481 arg1 epitope 483:489 the precise binding epitope 463:489 the precise binding epitope 463:489 Despite their importance and broad use, the precise binding epitope has been determined for only a few of these antibodies. 28924016 4 30 theme synthetic 603:611 arg1 oligosaccharides 613:628 88 synthetic oligosaccharides 600:628 88 synthetic oligosaccharides 600:628 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 5 31 theme arabinogalactan- 760:775 arg1 antibodies 831:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 Our results reveal the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies. 28924016 6 32 theme glycosyl 947:954 arg1 hydrolases 956:965 specific glycosyl hydrolases 938:965 specific glycosyl hydrolases 938:965 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 6 33 theme cell 1005:1008 arg1 analyses 1015:1022 immunological cell wall analyses 991:1022 immunological cell wall analyses 991:1022 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 4 34 theme cell 675:678 arg1 antibodies 701:710 plant cell wall glycan-directed antibodies 669:710 plant cell wall glycan-directed antibodies 669:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 6 35 theme specific 938:945 arg1 hydrolases 956:965 specific glycosyl hydrolases 938:965 specific glycosyl hydrolases 938:965 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 5 36 theme binding 736:742 arg1 epitopes 744:751 the binding epitopes 732:751 the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 732:840 Our results reveal the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies. 28924016 2 37 theme cell 340:343 arg1 components 350:359 plant cell wall components 334:359 plant cell wall components 334:359 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 5 38 theme rhamnogalacturonan- 778:796 arg1 antibodies 831:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies 757:840 Our results reveal the binding epitopes for 78 arabinogalactan-, rhamnogalacturonan-, xylan-, and xyloglucan-directed antibodies. 28924016 1 39 dep laboratories 250:261 arg1 worldwide 263:271 worldwide 263:271 different laboratories worldwide 240:271 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 2 40 theme plant 334:338 arg1 components 350:359 plant cell wall components 334:359 plant cell wall components 334:359 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 4 41 theme wall 680:683 arg1 antibodies 701:710 plant cell wall glycan-directed antibodies 669:710 plant cell wall glycan-directed antibodies 669:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 6 42 from information 1068:1078 arg1 glycans 1099:1105 plant cell wall glycans 1083:1105 plant cell wall glycans with unprecedented molecular precision 1083:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 4 43 theme antibodies 701:710 arg1 epitopes 657:664 the epitopes 653:664 the epitopes of plant cell wall glycan-directed antibodies 653:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 2 44 used used 302:305 arg2 antibodies 280:289 These antibodies 274:289 These antibodies 274:289 These antibodies are widely used to identify differences in plant cell wall components in mutants, organ and tissue types, and developmental stages. 28924016 1 45 theme plant 205:209 arg1 polysaccharides 221:235 plant cell wall polysaccharides 205:235 plant cell wall polysaccharides 205:235 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 6 46 theme structural 1057:1066 arg1 information 1068:1078 structural information 1057:1078 structural information on plant cell wall glycans with unprecedented molecular precision 1057:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 4 47 theme glycan-directed 685:699 arg1 antibodies 701:710 plant cell wall glycan-directed antibodies 669:710 plant cell wall glycan-directed antibodies 669:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 1 48 theme cell 211:214 arg1 polysaccharides 221:235 plant cell wall polysaccharides 205:235 plant cell wall polysaccharides 205:235 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 0 49 theme Cell 63:66 arg1 Antibodies 89:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 3 50 theme broad 452:456 arg1 use 458:460 broad use 452:460 broad use 452:460 Despite their importance and broad use, the precise binding epitope has been determined for only a few of these antibodies. 28924016 6 51 theme unprecedented 1112:1124 arg1 precision 1136:1144 unprecedented molecular precision 1112:1144 unprecedented molecular precision 1112:1144 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 1 52 theme wall 216:219 arg1 polysaccharides 221:235 plant cell wall polysaccharides 205:235 plant cell wall polysaccharides 205:235 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 28924016 0 53 theme Plant 57:61 arg1 Antibodies 89:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 Plant Cell Wall Glycan-Directed Antibodies 57:98 A Synthetic Glycan Microarray Enables Epitope Mapping of Plant Cell Wall Glycan-Directed Antibodies. 28924016 6 54 theme individual 915:924 arg1 antibodies 926:935 individual antibodies 915:935 individual antibodies 915:935 We demonstrate that, with knowledge of the exact epitopes recognized by individual antibodies, specific glycosyl hydrolases can be implemented into immunological cell wall analyses, providing a framework to obtain structural information on plant cell wall glycans with unprecedented molecular precision. 28924016 4 55 theme glycan 568:573 arg1 microarray 575:584 a plant glycan microarray 560:584 a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies 560:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 4 56 theme plant 562:566 arg1 microarray 575:584 a plant glycan microarray 560:584 a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies 560:710 Here, we use a plant glycan microarray equipped with 88 synthetic oligosaccharides to comprehensively map the epitopes of plant cell wall glycan-directed antibodies. 28924016 1 57 theme polysaccharides 221:235 arg1 classes 194:200 most classes 189:200 most classes of plant cell wall polysaccharides 189:235 In the last three decades, more than 200 monoclonal antibodies have been raised against most classes of plant cell wall polysaccharides by different laboratories worldwide. 26116146 5 0 theme additional 525:534 arg1 linker-region 536:548 an additional linker-region 522:548 an additional linker-region 522:548 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 5 1 theme CAZy 645:648 arg1 database 650:657 the CAZy database 641:657 the CAZy database 641:657 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 0 2 from effect 4:9 arg1 degradation 59:69 substrate degradation 49:69 substrate degradation by the human chitotriosidase 49:98 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 6 3 theme enzyme 689:694 arg1 functionality 696:708 enzyme functionality 689:708 enzyme functionality 689:708 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 5 4 contain has 518:520 arg2 linker-region 536:548 an additional linker-region 522:548 an additional linker-region 522:548 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 5 4 contain has 518:520 arg1 HCHT50 511:516 HCHT50 511:516 HCHT50 511:516 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 12 5 theme high 1665:1668 arg1 degree 1670:1675 a high degree 1663:1675 a high degree of efficiency 1663:1689 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 10 6 theme subsites 1280:1287 arg1 surface 1297:1303 the positive subsites binding surface 1267:1303 the positive subsites binding surface 1267:1303 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 12 7 theme chitinases 1611:1620 arg1 features 1561:1568 features 1561:1568 features of exo-processive and endo-nonprocessive chitinases 1561:1620 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 8 8 from kinetics 983:990 arg1 degradation 1004:1014 chitosan degradation 995:1014 chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase 995:1088 Interestingly, both HCHT50 and HCHT39 show biphasic kinetics on chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase. 26116146 0 9 theme human 78:82 arg1 chitotriosidase 84:98 the human chitotriosidase 74:98 the human chitotriosidase 74:98 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 12 10 theme alleged 1709:1715 arg1 task 1731:1734 its alleged physiological task 1705:1734 its alleged physiological task of being a "complete" chitinolytic machinery by itself 1705:1789 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 10 11 theme positive 1271:1278 arg1 surface 1297:1303 the positive subsites binding surface 1267:1303 the positive subsites binding surface 1267:1303 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 4 12 with domain 471:476 arg1 fold 505:508 the (β/α)8 TIM barrel fold 483:508 the (β/α)8 TIM barrel fold 483:508 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 26116146 4 13 theme β/α 488:490 arg1 fold 505:508 the (β/α)8 TIM barrel fold 483:508 the (β/α)8 TIM barrel fold 483:508 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 26116146 5 14 theme CBM 624:626 arg1 family 628:633 CBM family 14 624:636 CBM family 14 in the CAZy database 624:657 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 6 15 theme further 668:674 arg1 insight 676:682 further insight 668:682 further insight into enzyme functionality 668:708 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 1 16 theme active 144:149 arg1 chitinases 181:190 two active glycoside hydrolase family 18 chitinases 140:190 two active glycoside hydrolase family 18 chitinases produced by humans 140:209 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 12 17 theme efficiency 1680:1689 arg1 degree 1670:1675 a high degree 1663:1675 a high degree of efficiency 1663:1689 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 1 18 theme glycoside 151:159 arg1 chitinases 181:190 two active glycoside hydrolase family 18 chitinases 140:190 two active glycoside hydrolase family 18 chitinases produced by humans 140:209 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 6 19 theme weight 844:849 arg1 chitosans 851:859 chitin and high molecular weight chitosans 818:859 chitin and high molecular weight chitosans 818:859 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 10 20 dep CBM 1253:1255 arg1 extending 1257:1265 extending 1257:1265 extending the positive subsites binding surface 1257:1303 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 10 20 dep CBM 1253:1255 arg1 promoting 1319:1327 promoting 1319:1327 therefore promoting transglycosylation 1309:1346 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 1 21 theme hydrolase 161:169 arg1 chitinases 181:190 two active glycoside hydrolase family 18 chitinases 140:190 two active glycoside hydrolase family 18 chitinases produced by humans 140:209 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 4 22 theme catalytic 461:469 arg1 domain 471:476 a catalytic domain 459:476 a catalytic domain with the (β/α)8 TIM barrel fold 459:508 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 26116146 4 22 theme catalytic 461:469 arg1 Common 431:436 Common 431:436 Common for both isoforms 431:454 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 26116146 11 23 theme processivity 1453:1464 arg1 degree 1443:1448 a similarly low degree 1427:1448 a similarly low degree of processivity 1427:1464 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 10 24 theme increased 1196:1204 arg1 activity 1225:1232 increased transglycosylation activity 1196:1232 increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation 1196:1346 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 2 25 theme system 341:346 arg1 responses 310:318 the anti-parasite responses 292:318 the anti-parasite responses of the innate immune system 292:346 The enzyme is associated with several diseases and is thought to play a role in the anti-parasite responses of the innate immune system. 26116146 6 26 theme molecular 834:842 arg1 weight 844:849 high molecular weight 829:849 high molecular weight 829:849 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 12 27 theme chitinolytic 1758:1769 arg1 machinery 1771:1779 a "complete" chitinolytic machinery 1745:1779 a "complete" chitinolytic machinery 1745:1779 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 11 28 theme low 1439:1441 arg1 degree 1443:1448 a similarly low degree 1427:1448 a similarly low degree of processivity 1427:1464 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 10 29 theme binding 1289:1295 arg1 surface 1297:1303 the positive subsites binding surface 1267:1303 the positive subsites binding surface 1267:1303 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 6 30 theme high 829:832 arg1 weight 844:849 high molecular weight 829:849 high molecular weight 829:849 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 12 31 theme " 1756:1756 arg1 machinery 1771:1779 a "complete" chitinolytic machinery 1745:1779 a "complete" chitinolytic machinery 1745:1779 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 6 32 theme CBM 743:745 arg1 insight 676:682 further insight 668:682 further insight into enzyme functionality 668:708 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 6 32 theme CBM 743:745 arg1 effect 729:734 the effect 725:734 the effect of the CBM 725:745 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 1 33 theme Human 101:105 arg1 HCHT 124:127 HCHT 124:127 HCHT 124:127 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 1 33 theme Human 101:105 arg1 chitotriosidase 107:121 Human chitotriosidase 101:121 Human chitotriosidase (HCHT) 101:128 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 1 34 theme chitinases 181:190 arg1 one 133:135 one 133:135 one 133:135 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 1 34 theme chitinases 181:190 arg1 chitinases 181:190 two active glycoside hydrolase family 18 chitinases 140:190 two active glycoside hydrolase family 18 chitinases produced by humans 140:209 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 0 35 theme binding 31:37 arg1 module 39:44 the carbohydrate binding module 14:44 the carbohydrate binding module 14:44 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 12 36 theme isoforms 1523:1530 arg1 features 1503:1510 functional and structural features 1477:1510 functional and structural features of the two isoforms 1477:1530 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 5 37 from family 628:633 arg1 database 650:657 the CAZy database 641:657 the CAZy database 641:657 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 1 38 theme family 171:176 arg1 chitinases 181:190 two active glycoside hydrolase family 18 chitinases 140:190 two active glycoside hydrolase family 18 chitinases produced by humans 140:209 Human chitotriosidase (HCHT) is one of two active glycoside hydrolase family 18 chitinases produced by humans. 26116146 11 39 with studies 1358:1364 arg1 chitosan 1387:1394 chitosan 1387:1394 chitosan 1387:1394 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 11 39 with studies 1358:1364 arg1 chitin 1376:1381 chitin 1376:1381 chitin 1376:1381 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 0 40 theme carbohydrate 18:29 arg1 module 39:44 the carbohydrate binding module 14:44 the carbohydrate binding module 14:44 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 6 41 theme chitin 818:823 arg1 chitosans 851:859 chitin and high molecular weight chitosans 818:859 chitin and high molecular weight chitosans 818:859 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 8 42 theme second 1077:1082 arg1 phase 1084:1088 the second phase 1073:1088 the second phase 1073:1088 Interestingly, both HCHT50 and HCHT39 show biphasic kinetics on chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase. 26116146 12 43 theme complete 1748:1755 arg1 machinery 1771:1779 a "complete" chitinolytic machinery 1745:1779 a "complete" chitinolytic machinery 1745:1779 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 12 44 theme physiological 1717:1729 arg1 task 1731:1734 its alleged physiological task 1705:1734 its alleged physiological task of being a "complete" chitinolytic machinery by itself 1705:1789 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 12 45 theme structural 1492:1501 arg1 features 1503:1510 functional and structural features 1477:1510 functional and structural features of the two isoforms 1477:1530 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 11 46 contain have 1422:1425 arg1 isoforms 1413:1420 both isoforms 1408:1420 both isoforms 1408:1420 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 11 46 contain have 1422:1425 arg2 degree 1443:1448 a similarly low degree 1427:1448 a similarly low degree of processivity 1427:1464 Finally, studies with both chitin and chitosan showed that both isoforms have a similarly low degree of processivity. 26116146 8 47 theme chitosan 995:1002 arg1 degradation 1004:1014 chitosan degradation 995:1014 chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase 995:1088 Interestingly, both HCHT50 and HCHT39 show biphasic kinetics on chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase. 26116146 0 48 theme module 39:44 arg1 effect 4:9 The effect 0:9 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase 0:98 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 8 49 theme biphasic 974:981 arg1 kinetics 983:990 biphasic kinetics 974:990 biphasic kinetics on chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase 974:1088 Interestingly, both HCHT50 and HCHT39 show biphasic kinetics on chitosan degradation where HCHT50 is faster initially and HCHT39 is faster in the second phase. 26116146 9 50 theme different 1128:1136 arg1 distributions 1147:1159 distinctly different oligomer distributions 1117:1159 distinctly different oligomer distributions 1117:1159 Moreover, HCHT50 produces distinctly different oligomer distributions than HCHT39. 26116146 2 51 theme anti-parasite 296:308 arg1 responses 310:318 the anti-parasite responses 292:318 the anti-parasite responses of the innate immune system 292:346 The enzyme is associated with several diseases and is thought to play a role in the anti-parasite responses of the innate immune system. 26116146 6 52 theme catalytic 794:802 arg1 properties 804:813 their catalytic properties 788:813 their catalytic properties 788:813 To gain further insight into enzyme functionality and especially the effect of the CBM, we expressed both isoforms and compared their catalytic properties on chitin and high molecular weight chitosans. 26116146 5 53 theme C-terminal 565:574 arg1 CBM 605:607 CBM 605:607 CBM 605:607 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 5 53 theme C-terminal 565:574 arg1 module 597:602 a C-terminal carbohydrate-binding module 563:602 a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database 563:657 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 9 54 theme oligomer 1138:1145 arg1 distributions 1147:1159 distinctly different oligomer distributions 1117:1159 distinctly different oligomer distributions 1117:1159 Moreover, HCHT50 produces distinctly different oligomer distributions than HCHT39. 26116146 2 55 theme several 242:248 arg1 diseases 250:257 several diseases 242:257 several diseases 242:257 The enzyme is associated with several diseases and is thought to play a role in the anti-parasite responses of the innate immune system. 26116146 10 56 theme transglycosylation 1206:1223 arg1 activity 1225:1232 increased transglycosylation activity 1196:1232 increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation 1196:1346 This is likely due to increased transglycosylation activity for HCHT50 due the CBM extending the positive subsites binding surface and therefore promoting transglycosylation. 26116146 3 57 theme 39 406:407 arg1 kDa 409:411 kDa 409:411 kDa 409:411 HCHT occurs in two isoforms, one 50 kDa (HCHT50) and one 39 kDa variant (HCHT39). 26116146 12 58 with line 1695:1698 arg1 task 1731:1734 its alleged physiological task 1705:1734 its alleged physiological task of being a "complete" chitinolytic machinery by itself 1705:1789 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 4 59 theme TIM 494:496 arg1 fold 505:508 the (β/α)8 TIM barrel fold 483:508 the (β/α)8 TIM barrel fold 483:508 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 26116146 5 60 theme carbohydrate-binding 576:595 arg1 CBM 605:607 CBM 605:607 CBM 605:607 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 5 60 theme carbohydrate-binding 576:595 arg1 module 597:602 a C-terminal carbohydrate-binding module 563:602 a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database 563:657 HCHT50 has an additional linker-region, followed by a C-terminal carbohydrate-binding module (CBM) classified as CBM family 14 in the CAZy database. 26116146 3 61 theme kDa 409:411 arg1 variant 413:419 one 39 kDa variant 402:419 one 39 kDa variant (HCHT39) 402:428 HCHT occurs in two isoforms, one 50 kDa (HCHT50) and one 39 kDa variant (HCHT39). 26116146 3 61 theme kDa 409:411 arg1 isoforms 368:375 two isoforms 364:375 two isoforms 364:375 HCHT occurs in two isoforms, one 50 kDa (HCHT50) and one 39 kDa variant (HCHT39). 26116146 3 61 theme kDa 409:411 arg1 HCHT39 422:427 HCHT39 422:427 HCHT39 422:427 HCHT occurs in two isoforms, one 50 kDa (HCHT50) and one 39 kDa variant (HCHT39). 26116146 12 62 theme exo-processive 1573:1586 arg1 chitinases 1611:1620 exo-processive and endo-nonprocessive chitinases 1573:1620 exo-processive and endo-nonprocessive chitinases 1573:1620 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 2 63 theme immune 334:339 arg1 system 341:346 the innate immune system 323:346 the innate immune system 323:346 The enzyme is associated with several diseases and is thought to play a role in the anti-parasite responses of the innate immune system. 26116146 12 64 theme unusual 1640:1646 arg1 CBM14 1648:1652 the somewhat unusual CBM14 to reach a high degree of efficiency 1627:1689 the somewhat unusual CBM14 to reach a high degree of efficiency 1627:1689 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 12 65 theme functional 1477:1486 arg1 features 1503:1510 functional and structural features 1477:1510 functional and structural features of the two isoforms 1477:1530 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 0 66 theme substrate 49:57 arg1 degradation 59:69 substrate degradation 49:69 substrate degradation by the human chitotriosidase 49:98 The effect of the carbohydrate binding module on substrate degradation by the human chitotriosidase. 26116146 2 67 theme innate 327:332 arg1 system 341:346 the innate immune system 323:346 the innate immune system 323:346 The enzyme is associated with several diseases and is thought to play a role in the anti-parasite responses of the innate immune system. 26116146 12 68 theme endo-nonprocessive 1592:1609 arg1 chitinases 1611:1620 exo-processive and endo-nonprocessive chitinases 1573:1620 exo-processive and endo-nonprocessive chitinases 1573:1620 Combining functional and structural features of the two isoforms, it seems that HCHT combines features of exo-processive and endo-nonprocessive chitinases with the somewhat unusual CBM14 to reach a high degree of efficiency, in line with its alleged physiological task of being a "complete" chitinolytic machinery by itself. 26116146 4 69 theme barrel 498:503 arg1 fold 505:508 the (β/α)8 TIM barrel fold 483:508 the (β/α)8 TIM barrel fold 483:508 Common for both isoforms is a catalytic domain with the (β/α)8 TIM barrel fold. 27594301 0 0 theme cell-killing 94:105 arg1 activity 107:114 enhanced cell-killing activity 85:114 enhanced cell-killing activity 85:114 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 3 1 theme resulting 344:352 arg1 antibody 354:361 The resulting antibody 340:361 The resulting antibody 340:361 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 7 2 theme enhanced 1280:1287 arg1 activity 1302:1309 enhanced cell killing activity 1280:1309 enhanced cell killing activity 1280:1309 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 6 3 theme glycosylation 1104:1116 arg1 precursors 1084:1093 biosynthetic precursors 1071:1093 biosynthetic precursors of human glycosylation 1071:1116 In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. 27594301 4 4 theme typical 664:670 arg1 counterparts 678:689 the typical human counterparts 660:689 the typical human counterparts 660:689 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 0 5 theme enhanced 85:92 arg1 activity 107:114 enhanced cell-killing activity 85:114 enhanced cell-killing activity 85:114 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 6 6 theme terminal 1016:1023 arg1 structures 1033:1042 terminal glucose structures 1016:1042 terminal glucose structures 1016:1042 In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. 27594301 7 7 theme cell 1289:1292 arg1 activity 1302:1309 enhanced cell killing activity 1280:1309 enhanced cell killing activity 1280:1309 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 5 8 theme deposited 821:829 arg1 structures 839:848 deposited crystal structures 821:848 deposited crystal structures 821:848 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 0 9 gly glycoforms 63:72 arg1 antibody 54:61 unusual antibody glycoforms 46:72 unusual antibody glycoforms displaying enhanced cell-killing activity 46:114 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 3 10 theme unusual 454:460 arg1 glycosylation 471:483 unusual N-linked glycosylation 454:483 unusual N-linked glycosylation 454:483 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 5 11 theme crystal 831:837 arg1 structures 839:848 deposited crystal structures 821:848 deposited crystal structures 821:848 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 2 12 theme ciliate 306:312 arg1 Tetrahymena 315:325 the unicellular ciliate, Tetrahymena 290:325 Tetrahymena 315:325 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 4 13 theme human 672:676 arg1 counterparts 678:689 the typical human counterparts 660:689 the typical human counterparts 660:689 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 3 14 theme antibody-dependent 385:402 arg1 cytotoxicity 418:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 2 15 theme unicellular 294:304 arg1 Tetrahymena 315:325 the unicellular ciliate, Tetrahymena 290:325 Tetrahymena 315:325 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 2 16 from production 227:236 arg1 thermophila 327:337 the unicellular ciliate, Tetrahymena thermophila 290:337 the unicellular ciliate, Tetrahymena thermophila 290:337 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 1 17 theme antibody 182:189 arg1 therapeutics 191:202 antibody therapeutics 182:202 antibody therapeutics 182:202 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 3 18 theme cell-mediated 404:416 arg1 cytotoxicity 418:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 4 19 dep afucosylated 552:563 arg1 oligomannose-type 566:582 oligomannose-type 566:582 oligomannose-type 566:582 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 1 20 theme therapeutics 191:202 arg1 optimization 166:177 the optimization 162:177 the optimization of antibody therapeutics 162:202 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 7 21 theme monoclonal 1220:1229 arg1 antibodies 1231:1240 monoclonal antibodies 1220:1240 monoclonal antibodies exhibiting glycosylation 1220:1265 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 1 22 from parameter 149:157 arg1 optimization 166:177 the optimization 162:177 the optimization of antibody therapeutics 162:202 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 3 23 theme N-linked 462:469 arg1 glycosylation 471:483 unusual N-linked glycosylation 454:483 unusual N-linked glycosylation 454:483 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 5 24 theme native 974:979 arg1 glycans 994:1000 native complex-type glycans 974:1000 native complex-type glycans 974:1000 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 0 25 theme Antibody 0:7 arg1 production 9:18 Antibody production 0:18 Antibody production using a ciliate 0:34 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 6 26 theme human 1098:1102 arg1 glycosylation 1104:1116 human glycosylation 1098:1116 human glycosylation 1098:1116 In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. 27594301 7 27 theme antibodies 1231:1240 arg1 production 1206:1215 large-scale production 1194:1215 large-scale production of monoclonal antibodies exhibiting glycosylation 1194:1265 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 4 28 theme intermediates 753:765 arg1 fragments 730:738 fragments 730:738 fragments of metabolic intermediates observed in human glycoproteins 730:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 1 29 theme Antibody 117:124 arg1 parameter 149:157 a key parameter 143:157 a key parameter in the optimization of antibody therapeutics 143:202 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 1 29 theme Antibody 117:124 arg1 glycosylation 126:138 Antibody glycosylation 117:138 Antibody glycosylation 117:138 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 5 30 theme Fc 936:937 arg1 domain 939:944 the Fc domain 932:944 the Fc domain that closely mimic those of native complex-type glycans 932:1000 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 2 31 theme Tetrahymena 315:325 arg1 thermophila 327:337 the unicellular ciliate, Tetrahymena thermophila 290:337 the unicellular ciliate, Tetrahymena thermophila 290:337 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 6 32 theme glucose 1025:1031 arg1 structures 1033:1042 terminal glucose structures 1016:1042 terminal glucose structures 1016:1042 In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. 27594301 5 33 theme ciliate 871:877 arg1 glycans 879:885 the ciliate glycans 867:885 the ciliate glycans 867:885 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 6 34 theme biosynthetic 1071:1082 arg1 precursors 1084:1093 biosynthetic precursors 1071:1093 biosynthetic precursors of human glycosylation 1071:1116 In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. 27594301 4 35 theme spectrometric 520:532 arg1 analysis 534:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 2 36 theme anti-cancer 245:255 arg1 rituximab 277:285 the anti-cancer monoclonal antibody rituximab 241:285 the anti-cancer monoclonal antibody rituximab 241:285 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 5 37 theme complex-type 981:992 arg1 glycans 994:1000 native complex-type glycans 974:1000 native complex-type glycans 974:1000 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 7 38 theme killing 1294:1300 arg1 activity 1302:1309 enhanced cell killing activity 1280:1309 enhanced cell killing activity 1280:1309 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 4 39 theme metabolic 743:751 arg1 intermediates 753:765 metabolic intermediates 743:765 metabolic intermediates observed in human glycoproteins 743:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 7 40 theme large-scale 1194:1204 arg1 production 1206:1215 large-scale production 1194:1215 large-scale production of monoclonal antibodies exhibiting glycosylation 1194:1265 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 4 41 theme afucosylated 552:563 arg1 glycans 584:590 afucosylated, oligomannose-type glycans 552:590 afucosylated, oligomannose-type glycans 552:590 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 3 42 link N-linked 462:469 arg1 glycosylation 471:483 unusual N-linked glycosylation 454:483 unusual N-linked glycosylation 454:483 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 7 43 theme ciliate-based 1144:1156 arg1 systems 1169:1175 ciliate-based expression systems 1144:1175 ciliate-based expression systems 1144:1175 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 0 44 theme antibody 54:61 arg1 glycoforms 63:72 unusual antibody glycoforms 46:72 unusual antibody glycoforms displaying enhanced cell-killing activity 46:114 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 5 45 theme protein-carbohydrate 893:912 arg1 interactions 914:925 protein-carbohydrate interactions 893:925 protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans 893:1000 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 4 46 located observed 767:774 arg1 glycoproteins 785:797 human glycoproteins 779:797 human glycoproteins 779:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 46 located observed 767:774 arg2 intermediates 753:765 metabolic intermediates 743:765 metabolic intermediates observed in human glycoproteins 743:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 7 47 theme expression 1158:1167 arg1 systems 1169:1175 ciliate-based expression systems 1144:1175 ciliate-based expression systems 1144:1175 Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity. 27594301 0 48 theme unusual 46:52 arg1 glycoforms 63:72 unusual antibody glycoforms 46:72 unusual antibody glycoforms displaying enhanced cell-killing activity 46:114 Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity. 27594301 4 49 theme isomeric 622:629 arg1 whole 605:609 whole 605:609 whole 605:609 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 49 theme isomeric 622:629 arg1 structures 631:640 isomeric structures 622:640 isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins 622:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 50 theme chromatographic 495:509 arg1 analysis 534:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 51 gly glycoproteins 785:797 arg1 glycoproteins 785:797 human glycoproteins 779:797 human glycoproteins 779:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 52 theme Detailed 486:493 arg1 analysis 534:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 1 53 theme key 145:147 arg1 parameter 149:157 a key parameter 143:157 a key parameter in the optimization of antibody therapeutics 143:202 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 1 53 theme key 145:147 arg1 glycosylation 126:138 Antibody glycosylation 117:138 Antibody glycosylation 117:138 Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. 27594301 5 54 with interactions 914:925 arg1 domain 939:944 the Fc domain 932:944 the Fc domain that closely mimic those of native complex-type glycans 932:1000 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 5 55 theme structures 839:848 arg1 analysis 809:816 the analysis 805:816 the analysis of deposited crystal structures 805:848 From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. 27594301 4 56 theme mass 515:518 arg1 analysis 534:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis 486:541 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 2 57 theme rituximab 277:285 arg1 production 227:236 the production 223:236 the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila 223:337 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 2 58 theme antibody 268:275 arg1 rituximab 277:285 the anti-cancer monoclonal antibody rituximab 241:285 the anti-cancer monoclonal antibody rituximab 241:285 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 27594301 4 59 theme human 779:783 arg1 glycoproteins 785:797 human glycoproteins 779:797 human glycoproteins 779:797 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 4 60 gly afucosylated 552:563 arg1 glycans 584:590 afucosylated, oligomannose-type glycans 552:590 afucosylated, oligomannose-type glycans 552:590 Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. 27594301 3 61 theme enhanced 376:383 arg1 cytotoxicity 418:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 enhanced antibody-dependent cell-mediated cytotoxicity 376:429 The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. 27594301 2 62 theme monoclonal 257:266 arg1 rituximab 277:285 the anti-cancer monoclonal antibody rituximab 241:285 the anti-cancer monoclonal antibody rituximab 241:285 Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. 29031045 2 0 theme studied 301:307 arg1 subclass 313:320 the most extensively studied IgG subclass 280:320 the most extensively studied IgG subclass 280:320 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 2 0 theme studied 301:307 arg1 IgG1 272:275 IgG1 272:275 IgG1 272:275 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 1 1 theme unique 193:198 arg1 differences 200:210 unique differences 193:210 unique differences that result in subclass-specific effector functions 193:262 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 2 from subclasses 108:117 arg1 humans 122:127 humans 122:127 humans 122:127 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 3 3 theme binding 488:494 arg1 profile 496:502 a similar binding profile 478:502 a similar binding profile to Fcγ receptors 478:519 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 9 4 gly glycoforms 1574:1583 arg1 Fc 1571:1572 IgG3 Fc glycoforms 1566:1583 IgG3 Fc glycoforms 1566:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 9 4 gly glycoforms 1574:1583 arg1 IgG3 1566:1569 IgG3 Fc glycoforms 1566:1583 IgG3 Fc glycoforms 1566:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 9 5 theme binding 1545:1551 arg1 studies 1553:1559 binding studies 1545:1559 binding studies with IgG3 Fc glycoforms 1545:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 7 6 theme neonatal 1309:1316 arg1 FcRn 1331:1334 FcRn 1331:1334 FcRn 1331:1334 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 7 6 theme neonatal 1309:1316 arg1 receptor 1321:1328 the neonatal Fc receptor 1305:1328 the neonatal Fc receptor (FcRn) 1305:1335 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 9 7 theme Fc 1571:1572 arg1 glycoforms 1574:1583 IgG3 Fc glycoforms 1566:1583 IgG3 Fc glycoforms 1566:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 7 8 theme protein 1291:1297 arg1 G 1299:1299 protein G 1291:1299 protein G 1291:1299 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 5 9 from subclasses 949:958 arg1 structures 919:928 published structures 909:928 published structures from the other IgG subclasses 909:958 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 5 10 theme human 785:789 arg1 Fc 796:797 human IgG3 Fc 785:797 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 5 11 theme Fc 796:797 arg1 glycoform 838:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 7 12 from presence 1136:1143 arg1 IgG3 1156:1159 IgG3 1156:1159 IgG3 1156:1159 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 4 13 theme C1q 644:646 arg1 binding 659:665 C1q complement binding 644:665 C1q complement binding 644:665 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 3 14 theme stronger 525:532 arg1 activation 534:543 stronger activation 525:543 stronger activation of complement 525:557 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 8 15 theme IgG3 1342:1345 arg1 structure 1350:1358 The IgG3 Fc structure 1338:1358 The IgG3 Fc structure 1338:1358 The IgG3 Fc structure helps to explain some of these differences. 29031045 0 16 theme IgG3 59:62 arg1 Fc 64:65 human IgG3 Fc 53:65 human IgG3 Fc 53:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 1 17 contain have 188:191 arg1 subclasses 108:117 four subclasses 103:117 four subclasses in humans: IgG1, IgG2, IgG3 and IgG4 103:154 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 17 contain have 188:191 arg2 differences 200:210 unique differences 193:210 unique differences that result in subclass-specific effector functions 193:262 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 9 18 located observed 1442:1449 arg2 contacts 1433:1440 protein-glycan contacts 1418:1440 protein-glycan contacts observed in the crystal structure 1418:1474 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 9 18 located observed 1442:1449 arg1 structure 1466:1474 the crystal structure 1454:1474 the crystal structure 1454:1474 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 7 19 theme R435 1148:1151 arg1 presence 1136:1143 the presence 1132:1143 the presence of R435 in IgG3 (and H435 in the other IgG subclasses) 1132:1198 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 4 20 theme Fc 698:699 arg1 conformation 701:712 optimal Fc conformation 690:712 optimal Fc conformation 690:712 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 9 21 theme protein-glycan 1418:1431 arg1 contacts 1433:1440 protein-glycan contacts 1418:1440 protein-glycan contacts observed in the crystal structure 1418:1474 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 2 22 theme subclasses 338:347 arg1 study 323:327 study 323:327 study of other subclasses 323:347 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 1 23 theme Immunoglobulin 68:81 arg1 IgG 86:88 IgG 86:88 IgG 86:88 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 23 theme Immunoglobulin 68:81 arg1 G 83:83 Immunoglobulin G 68:83 Immunoglobulin G (IgG) 68:89 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 7 24 theme IgG3-specific 1233:1245 arg1 properties 1247:1256 IgG3-specific properties 1233:1256 IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn) 1233:1335 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 0 25 theme Structural 0:9 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of the Man5 glycoform of human IgG3 Fc. 0:66 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 6 26 theme Fc 1000:1001 arg1 similar 1006:1012 similar 1006:1012 similar 1006:1012 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 6 26 theme Fc 1000:1001 arg1 structure 982:990 the overall structure 970:990 the overall structure of IgG3 Fc 970:1001 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 2 27 theme immune 384:389 arg1 function 391:398 overall immune function 376:398 overall immune function and for development of new therapeutics 376:438 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 10 28 theme specific 1690:1697 arg1 gain 1699:1702 specific gain 1690:1702 specific gain of function 1690:1714 Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function. 29031045 1 29 theme subclass-specific 227:243 arg1 functions 254:262 subclass-specific effector functions 227:262 subclass-specific effector functions 227:262 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 5 30 theme structural 884:893 arg1 features 895:902 its structural features 880:902 its structural features 880:902 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 2 31 theme new 423:425 arg1 therapeutics 427:438 new therapeutics 423:438 new therapeutics 423:438 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 0 32 theme Man5 35:38 arg1 glycoform 40:48 the Man5 glycoform 31:48 the Man5 glycoform of human IgG3 Fc 31:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 6 33 theme overall 974:980 arg1 similar 1006:1012 similar 1006:1012 similar 1006:1012 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 6 33 theme overall 974:980 arg1 structure 982:990 the overall structure 970:990 the overall structure of IgG3 Fc 970:1001 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 10 34 theme function 1707:1714 arg1 gain 1699:1702 specific gain 1690:1702 specific gain of function 1690:1714 Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function. 29031045 6 35 theme structural 1048:1057 arg1 perturbations 1059:1071 some structural perturbations 1043:1071 some structural perturbations based on sequence differences 1043:1101 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 0 36 gly glycoform 40:48 arg1 Fc 64:65 human IgG3 Fc 53:65 human IgG3 Fc 53:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 0 36 gly glycoform 40:48 arg1 Man5 35:38 the Man5 glycoform 31:48 the Man5 glycoform of human IgG3 Fc 31:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 7 37 theme other 1178:1182 arg1 subclasses 1188:1197 the other IgG subclasses 1174:1197 the other IgG subclasses 1174:1197 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 10 38 theme Fc 1605:1606 arg1 structure 1608:1616 this IgG3 Fc structure 1595:1616 this IgG3 Fc structure 1595:1616 Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function. 29031045 5 39 theme IgG 945:947 arg1 subclasses 949:958 the other IgG subclasses 935:958 the other IgG subclasses 935:958 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 6 40 theme other 1025:1029 arg1 subclasses 1031:1040 other subclasses 1025:1040 other subclasses 1025:1040 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 2 41 theme IgG 309:311 arg1 subclass 313:320 the most extensively studied IgG subclass 280:320 the most extensively studied IgG subclass 280:320 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 2 41 theme IgG 309:311 arg1 IgG1 272:275 IgG1 272:275 IgG1 272:275 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 3 42 theme similar 480:486 arg1 profile 496:502 a similar binding profile 478:502 a similar binding profile to Fcγ receptors 478:519 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 7 43 from H435 1166:1169 arg1 subclasses 1188:1197 the other IgG subclasses 1174:1197 the other IgG subclasses 1174:1197 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 5 44 gly glycosylated 772:783 arg1 glycoform 838:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 7 45 theme Fc 1318:1319 arg1 FcRn 1331:1334 FcRn 1331:1334 FcRn 1331:1334 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 7 45 theme Fc 1318:1319 arg1 receptor 1321:1328 the neonatal Fc receptor 1305:1328 the neonatal Fc receptor (FcRn) 1305:1335 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 4 46 gly glycosylated 583:594 arg1 subclasses 568:577 All IgG subclasses 560:577 All IgG subclasses 560:577 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 4 46 gly glycosylated 583:594 arg2 N297 599:602 N297 599:602 N297 599:602 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 3 47 theme Fcγ 507:509 arg1 receptors 511:519 Fcγ receptors 507:519 Fcγ receptors 507:519 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 5 48 theme GlcNAc2Man5 806:816 arg1 mannose 830:836 a GlcNAc2Man5 (Man5) high mannose 804:836 a GlcNAc2Man5 (Man5) high mannose 804:836 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 9 49 theme IgG3 1566:1569 arg1 glycoforms 1574:1583 IgG3 Fc glycoforms 1566:1583 IgG3 Fc glycoforms 1566:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 6 50 theme sequence 1082:1089 arg1 differences 1091:1101 sequence differences 1082:1101 sequence differences 1082:1101 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 10 51 theme further 1642:1648 arg1 studies 1650:1656 further studies 1642:1656 further studies aimed at engineering the Fc for specific gain of function 1642:1714 Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function. 29031045 5 52 theme glycosylated 772:783 arg1 glycoform 838:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 5 53 gly glycoform 838:846 arg1 Fc 796:797 human IgG3 Fc 785:797 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 5 54 theme IgG3 791:794 arg1 Fc 796:797 human IgG3 Fc 785:797 homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform 759:846 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 7 55 attach presence 1136:1143 arg2 R435 1148:1151 R435 1148:1151 R435 1148:1151 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 7 55 attach presence 1136:1143 arg1 IgG3 1156:1159 IgG3 1156:1159 IgG3 1156:1159 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 5 56 theme glycoform 838:846 arg1 structure 746:754 the crystal structure 734:754 the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution 734:865 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 8 57 theme Fc 1347:1348 arg1 structure 1350:1358 The IgG3 Fc structure 1338:1358 The IgG3 Fc structure 1338:1358 The IgG3 Fc structure helps to explain some of these differences. 29031045 7 58 theme protein 1280:1286 arg1 A 1288:1288 protein A 1280:1288 protein A 1280:1288 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 5 59 theme 1.8Å 851:854 arg1 resolution 856:865 1.8Å resolution 851:865 1.8Å resolution 851:865 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 4 60 theme complement 648:657 arg1 binding 659:665 C1q complement binding 644:665 C1q complement binding 644:665 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 0 61 theme Fc 64:65 arg1 glycoform 40:48 the Man5 glycoform 31:48 the Man5 glycoform of human IgG3 Fc 31:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 3 62 theme complement 548:557 arg1 activation 534:543 stronger activation 525:543 stronger activation of complement 525:557 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 3 62 theme complement 548:557 arg1 profile 496:502 a similar binding profile 478:502 a similar binding profile to Fcγ receptors 478:519 When compared to IgG1, IgG3 exhibits a similar binding profile to Fcγ receptors and stronger activation of complement. 29031045 5 63 theme Man5 819:822 arg1 mannose 830:836 a GlcNAc2Man5 (Man5) high mannose 804:836 a GlcNAc2Man5 (Man5) high mannose 804:836 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 7 64 theme related 1258:1264 arg1 properties 1247:1256 IgG3-specific properties 1233:1256 IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn) 1233:1335 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 5 65 theme high 825:828 arg1 mannose 830:836 a GlcNAc2Man5 (Man5) high mannose 804:836 a GlcNAc2Man5 (Man5) high mannose 804:836 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 2 66 theme therapeutics 427:438 arg1 development 408:418 development 408:418 development of new therapeutics 408:438 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 4 67 theme Fcγ 627:629 arg1 receptor 631:638 Fcγ receptor 627:638 Fcγ receptor 627:638 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 4 68 theme optimal 690:696 arg1 conformation 701:712 optimal Fc conformation 690:712 optimal Fc conformation 690:712 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 1 69 dep subclasses 108:117 arg1 IgG4 151:154 IgG4 151:154 IgG4 151:154 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 69 dep subclasses 108:117 arg1 IgG3 142:145 IgG3 142:145 IgG3 142:145 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 69 dep subclasses 108:117 arg1 IgG2 136:139 IgG2 136:139 IgG2 136:139 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 1 69 dep subclasses 108:117 arg1 IgG1 130:133 IgG1 130:133 IgG1 130:133 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 5 70 theme published 909:917 arg1 structures 919:928 published structures 909:928 published structures from the other IgG subclasses 909:958 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 2 71 theme other 332:336 arg1 subclasses 338:347 other subclasses 332:347 other subclasses 332:347 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 5 72 theme crystal 738:744 arg1 structure 746:754 the crystal structure 734:754 the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution 734:865 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 5 73 from resolution 856:865 arg1 structure 746:754 the crystal structure 734:754 the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution 734:865 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 9 74 theme crystal 1458:1464 arg1 structure 1466:1474 the crystal structure 1454:1474 the crystal structure 1454:1474 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 6 75 theme IgG3 995:998 arg1 Fc 1000:1001 IgG3 Fc 995:1001 IgG3 Fc 995:1001 Although the overall structure of IgG3 Fc is similar to that of other subclasses, some structural perturbations based on sequence differences were revealed. 29031045 2 76 theme overall 376:382 arg1 function 391:398 overall immune function 376:398 overall immune function and for development of new therapeutics 376:438 Though IgG1 is the most extensively studied IgG subclass, study of other subclasses is important to understand overall immune function and for development of new therapeutics. 29031045 5 77 with Fc 796:797 arg1 mannose 830:836 a GlcNAc2Man5 (Man5) high mannose 804:836 a GlcNAc2Man5 (Man5) high mannose 804:836 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 0 78 theme glycoform 40:48 arg1 characterization 11:26 Structural characterization 0:26 Structural characterization of the Man5 glycoform of human IgG3 Fc. 0:66 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 1 79 theme effector 245:252 arg1 functions 254:262 subclass-specific effector functions 227:262 subclass-specific effector functions 227:262 Immunoglobulin G (IgG) consists of four subclasses in humans: IgG1, IgG2, IgG3 and IgG4, which are highly conserved but have unique differences that result in subclass-specific effector functions. 29031045 7 80 dep IgG3 1156:1159 arg1 H435 1166:1169 H435 1166:1169 H435 1166:1169 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 0 81 theme human 53:57 arg1 Fc 64:65 human IgG3 Fc 53:65 human IgG3 Fc 53:65 Structural characterization of the Man5 glycoform of human IgG3 Fc. 29031045 7 82 theme IgG 1184:1186 arg1 subclasses 1188:1197 the other IgG subclasses 1174:1197 the other IgG subclasses 1174:1197 For instance, the presence of R435 in IgG3 (and H435 in the other IgG subclasses) has been implicated to result in IgG3-specific properties related to binding to protein A, protein G and the neonatal Fc receptor (FcRn). 29031045 9 83 theme Fcγ 1519:1521 arg1 receptors 1523:1531 Fcγ receptors 1519:1531 Fcγ receptors 1519:1531 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 9 84 theme IgG3 1501:1504 arg1 affinity 1506:1513 IgG3 affinity 1501:1513 IgG3 affinity for Fcγ receptors 1501:1531 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 9 85 with studies 1553:1559 arg1 glycoforms 1574:1583 IgG3 Fc glycoforms 1566:1583 IgG3 Fc glycoforms 1566:1583 Additionally, protein-glycan contacts observed in the crystal structure appear to correlate with IgG3 affinity for Fcγ receptors as shown by binding studies with IgG3 Fc glycoforms. 29031045 5 86 theme other 939:943 arg1 subclasses 949:958 the other IgG subclasses 935:958 the other IgG subclasses 935:958 We have determined the crystal structure of homogenously glycosylated human IgG3 Fc with a GlcNAc2Man5 (Man5) high mannose glycoform at 1.8Å resolution and compared its structural features with published structures from the other IgG subclasses. 29031045 4 87 theme IgG 564:566 arg1 subclasses 568:577 All IgG subclasses 560:577 All IgG subclasses 560:577 All IgG subclasses are glycosylated at N297, which is required for Fcγ receptor and C1q complement binding as well as maintaining optimal Fc conformation. 29031045 10 88 theme IgG3 1600:1603 arg1 structure 1608:1616 this IgG3 Fc structure 1595:1616 this IgG3 Fc structure 1595:1616 Finally, this IgG3 Fc structure provides a template for further studies aimed at engineering the Fc for specific gain of function. 24682362 7 0 from response 1883:1890 arg1 patients 1895:1902 patients 1895:1902 patients receiving therapeutic antibodies 1895:1935 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 4 1 theme labile 1108:1113 arg1 H3 1148:1149 a labile complementary-determining region H3 1106:1149 a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1106:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 4 2 theme region 1141:1146 arg1 H3 1148:1149 a labile complementary-determining region H3 1106:1149 a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1106:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 5 3 theme heterogeneous 1290:1302 arg1 mixture 1304:1310 a heterogeneous mixture 1288:1310 a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties 1288:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 7 4 theme allergic 1874:1881 arg1 response 1883:1890 allergic response 1874:1890 allergic response in patients receiving therapeutic antibodies 1874:1935 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 4 5 theme chain 1211:1215 arg1 glycosylation 1175:1187 significant glycosylation 1163:1187 significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1163:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 1 6 theme binding 192:198 arg1 fragment 200:207 the antigen binding fragment 180:207 the antigen binding fragment of mAb S25-26 180:221 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 7 theme Chlamydiaceae-specific 494:515 arg1 mAbs 517:520 previously characterized Chlamydiaceae-specific mAbs 469:520 previously characterized Chlamydiaceae-specific mAbs 469:520 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 2 8 theme Kdo 596:598 arg1 pocket 612:617 the terminal Kdo recognition pocket 583:617 the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies 583:673 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 0 9 theme chain 140:144 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 10 theme 3-deoxy-α-d-manno-oct-2-ulopyranosonic 355:392 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 10 theme 3-deoxy-α-d-manno-oct-2-ulopyranosonic 355:392 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 11 theme N-linked 1766:1773 arg1 glycan 1775:1780 an αGal-containing N-linked glycan 1747:1780 an αGal-containing N-linked glycan on a mAb variable heavy chain 1747:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 7 12 theme three-dimensional 1716:1732 arg1 structure 1734:1742 the three-dimensional structure 1712:1742 the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain 1712:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 4 13 theme variable 1196:1203 arg1 chain 1211:1215 the variable heavy chain 1192:1215 the variable heavy chain 1192:1215 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 3 14 theme strict 906:911 arg1 specificity 913:923 strict specificity 906:923 strict specificity 906:923 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 2 15 theme germ-line 743:751 arg1 residues 753:760 germ-line residues 743:760 germ-line residues 743:760 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 1 16 theme germ-line-coded 412:426 arg1 paratope 428:435 a germ-line-coded paratope 410:435 a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen 410:573 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 6 17 theme electron 1657:1664 arg1 density 1666:1672 appropriate electron density 1645:1672 appropriate electron density 1645:1672 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 7 18 theme glycan 1775:1780 arg1 structure 1734:1742 the three-dimensional structure 1712:1742 the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain 1712:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 7 19 theme clinical 1826:1833 arg1 interest 1835:1842 potential clinical interest 1816:1842 potential clinical interest 1816:1842 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 2 20 theme trisaccharide 786:798 arg1 antigen 800:806 the entire trisaccharide antigen 775:806 the entire trisaccharide antigen 775:806 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 1 21 theme Kdo 336:338 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 21 theme Kdo 336:338 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 3 22 theme family-specific 933:947 arg1 antigen 963:969 the family-specific trisaccharide antigen 929:969 the family-specific trisaccharide antigen 929:969 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 3 23 theme high 885:888 arg1 affinity 893:900 its rare high μm affinity 876:900 its rare high μm affinity 876:900 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 0 24 theme antibodies 92:101 arg1 family 82:87 a family 80:87 a family of antibodies possessing an unusual variable heavy chain N-linked glycan 80:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 25 theme 2→4 340:342 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 25 theme 2→4 340:342 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 26 theme heavy 1800:1804 arg1 chain 1806:1810 a mAb variable heavy chain 1785:1810 a mAb variable heavy chain 1785:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 3 27 from Interest 842:849 arg1 S25-23 854:859 S25-23 854:859 S25-23 854:859 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 6 28 theme structures 1435:1444 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 28 theme structures 1435:1444 arg1 One 1411:1413 One 1411:1413 One 1411:1413 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 28 theme structures 1435:1444 arg1 Cetuximab 1521:1529 the therapeutic antibody Cetuximab 1496:1529 the therapeutic antibody Cetuximab 1496:1529 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 7 29 theme mAb 1787:1789 arg1 chain 1806:1810 a mAb variable heavy chain 1785:1810 a mAb variable heavy chain 1785:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 0 30 theme variable 125:132 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 31 theme Kdo 349:351 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 31 theme Kdo 349:351 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 0 32 theme antigen 69:75 arg1 recognition 12:22 Groove-type recognition 0:22 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 0:161 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 0 33 link N-linked 146:153 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 4 34 theme S25-26 1094:1099 arg1 forms 1085:1089 three unliganded forms 1068:1089 three unliganded forms of S25-26 1068:1099 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 0 35 theme Groove-type 0:10 arg1 recognition 12:22 Groove-type recognition 0:22 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 0:161 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 36 theme Chlamydiaceae 276:288 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 36 theme Chlamydiaceae 276:288 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 6 37 from structures 1581:1590 arg1 S25-26 1595:1600 S25-26 1595:1600 S25-26 1595:1600 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 3 38 theme related 990:996 arg1 S25-26 1007:1012 only the related antibody S25-26 981:1012 only the related antibody S25-26 981:1012 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 5 39 theme terminal 1383:1390 arg1 moieties 1401:1408 terminal αGal-Gal moieties 1383:1408 terminal αGal-Gal moieties 1383:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 6 40 theme antibody 1512:1519 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 40 theme antibody 1512:1519 arg1 Cetuximab 1521:1529 the therapeutic antibody Cetuximab 1496:1529 the therapeutic antibody Cetuximab 1496:1529 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 40 theme antibody 1512:1519 arg1 One 1411:1413 One 1411:1413 One 1411:1413 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 41 theme trisaccharide 306:318 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 41 theme trisaccharide 306:318 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 5 42 theme moieties 1401:1408 arg1 number 1373:1378 an unusually high number 1355:1378 an unusually high number of terminal αGal-Gal moieties 1355:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 0 43 theme lipopolysaccharide 50:67 arg1 antigen 69:75 chlamydiaceae-specific lipopolysaccharide antigen 27:75 chlamydiaceae-specific lipopolysaccharide antigen 27:75 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 44 theme Kdo 328:330 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 44 theme Kdo 328:330 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 4 45 theme forms 1085:1089 arg1 structures 1054:1063 The structures 1050:1063 The structures of three unliganded forms of S25-26 1050:1099 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 1 46 theme 2→8 332:334 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 46 theme 2→8 332:334 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 47 theme therapeutic 1914:1924 arg1 antibodies 1926:1935 therapeutic antibodies 1914:1935 therapeutic antibodies 1914:1935 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 1 48 theme identical 555:563 arg1 immunogen 565:573 the identical immunogen 551:573 the identical immunogen 551:573 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 4 49 from asparagine 1220:1229 arg1 Region 1247:1252 Framework Region 3 1237:1254 Framework Region 3 1237:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 1 50 theme Å 243:243 arg1 resolution 245:254 1.95 Å resolution 238:254 1.95 Å resolution 238:254 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 4 51 theme complementary-determining 1115:1139 arg1 H3 1148:1149 a labile complementary-determining region H3 1106:1149 a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1106:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 5 52 theme glycan 1273:1278 arg1 Analysis 1257:1264 Analysis 1257:1264 Analysis of the glycan 1257:1278 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 2 53 theme S25-2-type 653:662 arg1 antibodies 664:673 S25-2-type antibodies 653:673 S25-2-type antibodies 653:673 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 7 54 contain has 1812:1814 arg1 elucidation 1697:1707 The elucidation 1693:1707 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain 1693:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 7 54 contain has 1812:1814 arg2 interest 1835:1842 potential clinical interest 1816:1842 potential clinical interest 1816:1842 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 6 55 contain containing 1467:1476 arg1 mAbs 1462:1465 glycosylated mAbs 1449:1465 glycosylated mAbs containing these epitopes 1449:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 55 contain containing 1467:1476 arg2 epitopes 1484:1491 these epitopes 1478:1491 these epitopes 1478:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 7 56 from chain 1806:1810 arg1 structure 1734:1742 the three-dimensional structure 1712:1742 the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain 1712:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 2 57 theme strict 822:827 arg1 specificity 829:839 strict specificity 822:839 strict specificity 822:839 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 7 58 theme potential 1816:1824 arg1 interest 1835:1842 potential clinical interest 1816:1842 potential clinical interest 1816:1842 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 1 59 theme antigen 184:190 arg1 fragment 200:207 the antigen binding fragment 180:207 the antigen binding fragment of mAb S25-26 180:221 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 4 60 contain have 1101:1104 arg2 H3 1148:1149 a labile complementary-determining region H3 1106:1149 a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1106:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 4 60 contain have 1101:1104 arg1 structures 1054:1063 The structures 1050:1063 The structures of three unliganded forms of S25-26 1050:1099 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 6 61 theme glycosylated 1449:1460 arg1 mAbs 1462:1465 glycosylated mAbs 1449:1465 glycosylated mAbs containing these epitopes 1449:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 62 theme characterized 480:492 arg1 mAbs 517:520 previously characterized Chlamydiaceae-specific mAbs 469:520 previously characterized Chlamydiaceae-specific mAbs 469:520 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 6 63 theme significant 1608:1618 arg1 order 1620:1624 significant order 1608:1624 significant order in the glycan with appropriate electron density for nine residues 1608:1690 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 64 theme fragment 200:207 arg1 structure 167:175 The structure 163:175 The structure 163:175 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 65 link N-linked 1766:1773 arg1 glycan 1775:1780 an αGal-containing N-linked glycan 1747:1780 an αGal-containing N-linked glycan on a mAb variable heavy chain 1747:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 7 66 theme structure 1734:1742 arg1 elucidation 1697:1707 The elucidation 1693:1707 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain 1693:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 0 67 theme N-linked 146:153 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 68 theme mAb 212:214 arg1 fragment 200:207 the antigen binding fragment 180:207 the antigen binding fragment of mAb S25-26 180:221 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 2 69 theme terminal 587:594 arg1 pocket 612:617 the terminal Kdo recognition pocket 583:617 the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies 583:673 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 4 70 theme heavy 1205:1209 arg1 chain 1211:1215 the variable heavy chain 1192:1215 the variable heavy chain 1192:1215 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 6 71 theme structures 1581:1590 arg1 structures 1581:1590 the unliganded structures 1566:1590 the unliganded structures in S25-26 1566:1600 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 71 theme structures 1581:1590 arg1 one 1559:1561 one 1559:1561 one 1559:1561 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 4 72 gly glycosylation 1175:1187 arg1 asparagine 1220:1229 asparagine 85 1220:1232 asparagine 85 in Framework Region 3 1220:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 4 72 gly glycosylation 1175:1187 arg1 chain 1211:1215 the variable heavy chain 1192:1215 the variable heavy chain 1192:1215 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 4 73 from glycosylation 1175:1187 arg1 asparagine 1220:1229 asparagine 85 1220:1232 asparagine 85 in Framework Region 3 1220:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 6 74 with order 1620:1624 arg1 density 1666:1672 appropriate electron density 1645:1672 appropriate electron density 1645:1672 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 7 75 from glycan 1775:1780 arg1 chain 1806:1810 a mAb variable heavy chain 1785:1810 a mAb variable heavy chain 1785:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 5 76 contain contains 1346:1353 arg2 number 1373:1378 an unusually high number 1355:1378 an unusually high number of terminal αGal-Gal moieties 1355:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 5 76 contain contains 1346:1353 arg1 structure 1331:1339 a common root structure 1317:1339 a common root structure that contains an unusually high number of terminal αGal-Gal moieties 1317:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 2 77 theme recognition 600:610 arg1 pocket 612:617 the terminal Kdo recognition pocket 583:617 the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies 583:673 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 4 78 theme Framework 1237:1245 arg1 Region 1247:1252 Framework Region 3 1237:1254 Framework Region 3 1237:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 5 79 theme high 1368:1371 arg1 number 1373:1378 an unusually high number 1355:1378 an unusually high number of terminal αGal-Gal moieties 1355:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 6 80 theme appropriate 1645:1655 arg1 density 1666:1672 appropriate electron density 1645:1672 appropriate electron density 1645:1672 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 2 81 theme entire 779:784 arg1 antigen 800:806 the entire trisaccharide antigen 775:806 the entire trisaccharide antigen 775:806 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 5 82 theme root 1326:1329 arg1 structure 1331:1339 a common root structure 1317:1339 a common root structure that contains an unusually high number of terminal αGal-Gal moieties 1317:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 6 83 theme mAbs 1462:1465 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 3 84 theme trisaccharide 949:961 arg1 antigen 963:969 the family-specific trisaccharide antigen 929:969 the family-specific trisaccharide antigen 929:969 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 3 85 theme μm 890:891 arg1 affinity 893:900 its rare high μm affinity 876:900 its rare high μm affinity 876:900 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 0 86 theme unusual 117:123 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 6 87 from order 1620:1624 arg1 glycan 1633:1638 the glycan 1629:1638 the glycan 1629:1638 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 2 88 theme related 699:705 arg1 S25-23 707:712 the closely related S25-23 687:712 the closely related S25-23 687:712 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 3 89 theme rare 880:883 arg1 affinity 893:900 its rare high μm affinity 876:900 its rare high μm affinity 876:900 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 0 90 theme heavy 134:138 arg1 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 7 91 theme variable 1791:1798 arg1 chain 1806:1810 a mAb variable heavy chain 1785:1810 a mAb variable heavy chain 1785:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 6 92 theme reported 1426:1433 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 93 theme = 353:353 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 93 theme = 353:353 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 94 from structure 1734:1742 arg1 chain 1806:1810 a mAb variable heavy chain 1785:1810 a mAb variable heavy chain 1785:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 0 95 theme chlamydiaceae-specific 27:48 arg1 antigen 69:75 chlamydiaceae-specific lipopolysaccharide antigen 27:75 chlamydiaceae-specific lipopolysaccharide antigen 27:75 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 1 96 theme family-specific 290:304 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 96 theme family-specific 290:304 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 3 97 theme antibody 998:1005 arg1 S25-26 1007:1012 only the related antibody S25-26 981:1012 only the related antibody S25-26 981:1012 Interest in S25-23 was sparked by its rare high μm affinity and strict specificity for the family-specific trisaccharide antigen; however, only the related antibody S25-26 proved amenable to crystallization. 24682362 0 98 contain possessing 103:112 arg1 antibodies 92:101 antibodies 92:101 antibodies possessing an unusual variable heavy chain N-linked glycan 92:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 0 98 contain possessing 103:112 arg2 glycan 155:160 an unusual variable heavy chain N-linked glycan 114:160 an unusual variable heavy chain N-linked glycan 114:160 Groove-type recognition of chlamydiaceae-specific lipopolysaccharide antigen by a family of antibodies possessing an unusual variable heavy chain N-linked glycan. 24682362 5 99 theme αGal-Gal 1392:1399 arg1 moieties 1401:1408 terminal αGal-Gal moieties 1383:1408 terminal αGal-Gal moieties 1383:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 6 100 theme therapeutic 1500:1510 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 100 theme therapeutic 1500:1510 arg1 Cetuximab 1521:1529 the therapeutic antibody Cetuximab 1496:1529 the therapeutic antibody Cetuximab 1496:1529 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 6 100 theme therapeutic 1500:1510 arg1 One 1411:1413 One 1411:1413 One 1411:1413 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 101 theme antigen 320:326 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 101 theme antigen 320:326 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 7 102 theme αGal-containing 1750:1764 arg1 glycan 1775:1780 an αGal-containing N-linked glycan 1747:1780 an αGal-containing N-linked glycan on a mAb variable heavy chain 1747:1810 The elucidation of the three-dimensional structure of an αGal-containing N-linked glycan on a mAb variable heavy chain has potential clinical interest, as it has been implicated in allergic response in patients receiving therapeutic antibodies. 24682362 6 103 theme few 1422:1424 arg1 structures 1435:1444 the few reported structures 1418:1444 the few reported structures of glycosylated mAbs containing these epitopes 1418:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 4 104 theme unliganded 1074:1083 arg1 forms 1085:1089 three unliganded forms 1068:1089 three unliganded forms of S25-26 1068:1099 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 6 105 theme unliganded 1570:1579 arg1 structures 1581:1590 the unliganded structures 1566:1590 the unliganded structures in S25-26 1566:1600 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 1 106 with complex 259:265 arg1 acid 394:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid 349:397 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 1 106 with complex 259:265 arg1 Kdo 344:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo 272:346 the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid) 272:398 The structure of the antigen binding fragment of mAb S25-26, determined to 1.95 Å resolution in complex with the Chlamydiaceae family-specific trisaccharide antigen Kdo(2→8)Kdo(2→4)Kdo (Kdo = 3-deoxy-α-d-manno-oct-2-ulopyranosonic acid), displays a germ-line-coded paratope that differs significantly from previously characterized Chlamydiaceae-specific mAbs despite being raised against the identical immunogen. 24682362 4 107 theme adjacent 1151:1158 arg1 H3 1148:1149 a labile complementary-determining region H3 1106:1149 a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1106:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 6 108 gly glycosylated 1449:1460 arg1 mAbs 1462:1465 glycosylated mAbs 1449:1465 glycosylated mAbs containing these epitopes 1449:1491 One of the few reported structures of glycosylated mAbs containing these epitopes is the therapeutic antibody Cetuximab; however, unlike Cetuximab, one of the unliganded structures in S25-26 shows significant order in the glycan with appropriate electron density for nine residues. 24682362 2 109 dep and 808:810 arg1 so 812:813 so 812:813 so 812:813 Unlike the terminal Kdo recognition pocket that promotes cross-reactivity in S25-2-type antibodies, S25-26 and the closely related S25-23 utilize a groove composed of germ-line residues to recognize the entire trisaccharide antigen and so confer strict specificity. 24682362 4 110 theme significant 1163:1173 arg1 glycosylation 1175:1187 significant glycosylation 1163:1187 significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3 1163:1254 The structures of three unliganded forms of S25-26 have a labile complementary-determining region H3 adjacent to significant glycosylation of the variable heavy chain on asparagine 85 in Framework Region 3. 24682362 5 111 with mixture 1304:1310 arg1 structure 1331:1339 a common root structure 1317:1339 a common root structure that contains an unusually high number of terminal αGal-Gal moieties 1317:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 24682362 5 112 theme common 1319:1324 arg1 structure 1331:1339 a common root structure 1317:1339 a common root structure that contains an unusually high number of terminal αGal-Gal moieties 1317:1408 Analysis of the glycan reveals a heterogeneous mixture with a common root structure that contains an unusually high number of terminal αGal-Gal moieties. 28672164 8 0 theme N-glycans 1168:1176 arg1 presence 1133:1140 the presence 1129:1140 the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation 1129:1244 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 2 1 attach isolated 387:394 arg1 lucorum 407:413 Helix lucorum 401:413 Helix lucorum 401:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 2 1 attach isolated 387:394 arg2 hemocyanin 376:385 hemocyanin 376:385 hemocyanin isolated from Helix lucorum 376:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 7 2 theme carbohydrate 1003:1014 arg1 chains 1016:1021 Several carbohydrate chains 995:1021 Several carbohydrate chains in β-HlH 995:1030 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 4 3 theme glycans 609:615 arg1 mixture 598:604 a highly heterogeneous mixture 575:604 a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 575:669 Our study revealed a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1. 28672164 5 4 theme Asn-bound 831:839 arg1 GlcNAc 841:846 the Asn-bound GlcNAc 827:846 the Asn-bound GlcNAc 827:846 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 4 theme Asn-bound 831:839 arg1 residue 818:824 the innermost GlcNAc residue 797:824 the innermost GlcNAc residue (the Asn-bound GlcNAc) 797:847 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 2 5 theme β-HlH 367:371 arg1 N-glycans 331:339 N-glycans 331:339 N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum 331:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 1 6 theme clinical 277:284 arg1 studies 286:292 clinical studies 277:292 clinical studies 277:292 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 5 7 theme inner 726:730 arg1 core 732:735 the inner core 722:735 the inner core 722:735 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 4 8 theme composition 622:632 arg1 Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 634:669 composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 622:669 composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 622:669 Our study revealed a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1. 28672164 1 9 from interest 153:160 arg1 studies 286:292 clinical studies 277:292 clinical studies 277:292 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 2 10 theme first 303:307 arg1 time 309:312 the first time 299:312 the first time 299:312 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 3 11 theme mass 504:507 arg1 spectrometry 509:520 tandem mass spectrometry 497:520 tandem mass spectrometry using a Q-Trap mass spectrometer 497:553 In total, 32 different glycans were enzymatically liberated and characterized by tandem mass spectrometry using a Q-Trap mass spectrometer. 28672164 5 12 theme innermost 801:809 arg1 GlcNAc 841:846 the Asn-bound GlcNAc 827:846 the Asn-bound GlcNAc 827:846 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 12 theme innermost 801:809 arg1 residue 818:824 the innermost GlcNAc residue 797:824 the innermost GlcNAc residue (the Asn-bound GlcNAc) 797:847 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 6 13 contain contain 898:904 arg2 residue 923:929 a terminal MeHex residue 906:929 a terminal MeHex residue 906:929 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 6 13 contain contain 898:904 arg1 glycans 874:880 The glycans 870:880 The glycans of β-HlH 870:889 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 5 14 mod modified 710:717 arg1 chains 692:697 The oligosaccharide chains 672:697 The oligosaccharide chains 672:697 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 14 mod modified 710:717 arg3 xylose 752:757 β1-2-linked xylose 740:757 β1-2-linked xylose to β-mannose 740:770 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 15 theme oligosaccharide 676:690 arg1 chains 692:697 The oligosaccharide chains 672:697 The oligosaccharide chains 672:697 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 3 16 theme different 429:437 arg1 glycans 439:445 32 different glycans 426:445 32 different glycans 426:445 In total, 32 different glycans were enzymatically liberated and characterized by tandem mass spectrometry using a Q-Trap mass spectrometer. 28672164 1 17 theme specific 175:182 arg1 composition 199:209 their specific monosaccharide composition 169:209 their specific monosaccharide composition 169:209 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 5 18 theme GlcNAc 811:816 arg1 GlcNAc 841:846 the Asn-bound GlcNAc 827:846 the Asn-bound GlcNAc 827:846 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 18 theme GlcNAc 811:816 arg1 residue 818:824 the innermost GlcNAc residue 797:824 the innermost GlcNAc residue (the Asn-bound GlcNAc) 797:847 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 1 19 theme carbohydrate 70:81 arg1 structures 83:92 The carbohydrate structures 66:92 The carbohydrate structures of molluscan hemocyanins 66:117 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 7 20 contain possess 1074:1080 arg1 chains 1016:1021 Several carbohydrate chains 995:1021 Several carbohydrate chains in β-HlH 995:1030 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 7 20 contain possess 1074:1080 arg2 degree 1089:1094 a high degree 1082:1094 a high degree of methylation 1082:1109 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 1 21 theme monosaccharide 184:197 arg1 composition 199:209 their specific monosaccharide composition 169:209 their specific monosaccharide composition 169:209 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 8 22 theme structures 1201:1210 arg1 presence 1133:1140 the presence 1129:1140 the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation 1129:1244 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 8 23 theme bi-antennary 1155:1166 arg1 N-glycans 1168:1176 mono- and bi-antennary N-glycans 1145:1176 mono- and bi-antennary N-glycans as well as hybrid type structures 1145:1210 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 7 24 theme methylation 1099:1109 arg1 degree 1089:1094 a high degree 1082:1094 a high degree of methylation 1082:1109 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 2 25 theme Helix 401:405 arg1 lucorum 407:413 Helix lucorum 401:413 Helix lucorum 401:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 0 26 theme N-glycan 0:7 arg1 structures 9:18 N-glycan structures 0:18 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin 0:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 0 27 theme β-HlH 23:27 arg1 subunit 29:35 β-HlH subunit 23:35 β-HlH subunit of Helix lucorum hemocyanin 23:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 1 28 theme molluscan 97:105 arg1 hemocyanins 107:117 molluscan hemocyanins 97:117 molluscan hemocyanins 97:117 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 6 29 theme residues 979:986 arg1 residues 979:986 these residues 973:986 these residues 973:986 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 6 29 theme residues 979:986 arg1 two 951:953 two 951:953 two 951:953 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 6 30 theme MeHex 917:921 arg1 residue 923:929 a terminal MeHex residue 906:929 a terminal MeHex residue 906:929 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 1 31 theme hemocyanins 107:117 arg1 structures 83:92 The carbohydrate structures 66:92 The carbohydrate structures of molluscan hemocyanins 66:117 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 4 32 theme heterogeneous 584:596 arg1 mixture 598:604 a highly heterogeneous mixture 575:604 a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 575:669 Our study revealed a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1. 28672164 6 33 theme terminal 908:915 arg1 residue 923:929 a terminal MeHex residue 906:929 a terminal MeHex residue 906:929 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 0 34 theme subunit 29:35 arg1 structures 9:18 N-glycan structures 0:18 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin 0:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 7 35 theme high 1084:1087 arg1 degree 1089:1094 a high degree 1082:1094 a high degree of methylation 1082:1109 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 0 36 theme lucorum 46:52 arg1 hemocyanin 54:63 Helix lucorum hemocyanin 40:63 Helix lucorum hemocyanin 40:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 7 37 gly core-fucosylated 1036:1051 arg1 chains 1016:1021 Several carbohydrate chains 995:1021 Several carbohydrate chains in β-HlH 995:1030 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 5 38 link β1-2-linked 740:750 arg1 xylose 752:757 β1-2-linked xylose 740:757 β1-2-linked xylose to β-mannose 740:770 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 2 39 theme subunit 359:365 arg1 β-HlH 367:371 the structural subunit β-HlH 344:371 the structural subunit β-HlH of hemocyanin isolated from Helix lucorum 344:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 0 40 theme Helix 40:44 arg1 hemocyanin 54:63 Helix lucorum hemocyanin 40:63 Helix lucorum hemocyanin 40:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 5 41 theme β1-2-linked 740:750 arg1 xylose 752:757 β1-2-linked xylose 740:757 β1-2-linked xylose to β-mannose 740:770 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 3 42 theme Q-Trap 530:535 arg1 spectrometer 542:553 a Q-Trap mass spectrometer 528:553 a Q-Trap mass spectrometer 528:553 In total, 32 different glycans were enzymatically liberated and characterized by tandem mass spectrometry using a Q-Trap mass spectrometer. 28672164 2 43 theme structural 348:357 arg1 β-HlH 367:371 the structural subunit β-HlH 344:371 the structural subunit β-HlH of hemocyanin isolated from Helix lucorum 344:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 28672164 6 44 theme β-HlH 885:889 arg1 glycans 874:880 The glycans 870:880 The glycans of β-HlH 870:889 The glycans of β-HlH mainly contain a terminal MeHex residue; in some cases even two, three or four of these residues occur. 28672164 3 45 theme tandem 497:502 arg1 spectrometry 509:520 tandem mass spectrometry 497:520 tandem mass spectrometry using a Q-Trap mass spectrometer 497:553 In total, 32 different glycans were enzymatically liberated and characterized by tandem mass spectrometry using a Q-Trap mass spectrometer. 28672164 1 46 theme immunostimulatory 229:245 arg1 properties 247:256 their immunostimulatory properties 223:256 their immunostimulatory properties 223:256 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 8 47 theme type 1196:1199 arg1 structures 1201:1210 hybrid type structures 1189:1210 mono- and bi-antennary N-glycans as well as hybrid type structures 1145:1210 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 1 48 theme particular 142:151 arg1 interest 153:160 particular interest 142:160 particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies 142:292 The carbohydrate structures of molluscan hemocyanins have recently received particular interest due to their specific monosaccharide composition, as well as their immunostimulatory properties and application in clinical studies. 28672164 8 49 theme mono- 1145:1149 arg1 N-glycans 1168:1176 mono- and bi-antennary N-glycans 1145:1176 mono- and bi-antennary N-glycans as well as hybrid type structures 1145:1210 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 5 50 gly α1-6-fucosylation 776:792 arg1 GlcNAc 841:846 the Asn-bound GlcNAc 827:846 the Asn-bound GlcNAc 827:846 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 50 gly α1-6-fucosylation 776:792 arg1 residue 818:824 the innermost GlcNAc residue 797:824 the innermost GlcNAc residue (the Asn-bound GlcNAc) 797:847 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 5 51 theme residue 818:824 arg1 α1-6-fucosylation 776:792 α1-6-fucosylation 776:792 α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc) 776:847 The oligosaccharide chains are mostly modified at the inner core by β1-2-linked xylose to β-mannose, by α1-6-fucosylation of the innermost GlcNAc residue (the Asn-bound GlcNAc), and by methylation. 28672164 4 52 with glycans 609:615 arg1 Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 634:669 composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 622:669 composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1 622:669 Our study revealed a highly heterogeneous mixture of glycans with composition Hex3-7HexNAc2-5MeHex0-4Pent0-1Fuc0-1. 28672164 8 53 theme hybrid 1189:1194 arg1 structures 1201:1210 hybrid type structures 1189:1210 mono- and bi-antennary N-glycans as well as hybrid type structures 1145:1210 This study shows the presence of mono- and bi-antennary N-glycans as well as hybrid type structures with or without core-fucosylation. 28672164 3 54 theme mass 537:540 arg1 spectrometer 542:553 a Q-Trap mass spectrometer 528:553 a Q-Trap mass spectrometer 528:553 In total, 32 different glycans were enzymatically liberated and characterized by tandem mass spectrometry using a Q-Trap mass spectrometer. 28672164 7 55 from chains 1016:1021 arg1 β-HlH 1026:1030 β-HlH 1026:1030 β-HlH 1026:1030 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 0 56 theme hemocyanin 54:63 arg1 subunit 29:35 β-HlH subunit 23:35 β-HlH subunit of Helix lucorum hemocyanin 23:63 N-glycan structures of β-HlH subunit of Helix lucorum hemocyanin. 28672164 7 57 theme Several 995:1001 arg1 chains 1016:1021 Several carbohydrate chains 995:1021 Several carbohydrate chains in β-HlH 995:1030 Several carbohydrate chains in β-HlH are core-fucosylated without Xyl and also possess a high degree of methylation. 28672164 2 58 theme hemocyanin 376:385 arg1 β-HlH 367:371 the structural subunit β-HlH 344:371 the structural subunit β-HlH of hemocyanin isolated from Helix lucorum 344:413 For the first time, we investigated N-glycans of the structural subunit β-HlH of hemocyanin isolated from Helix lucorum. 25263869 5 0 theme lignocellulosic 706:720 arg1 materials 722:730 these lignocellulosic materials 700:730 these lignocellulosic materials 700:730 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 6 1 theme chemical 882:889 arg1 properties 891:900 the chemical properties 878:900 the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose 878:970 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 4 2 theme oxides 627:632 arg1 content 588:594 a higher content 579:594 a higher content of alkaline and alkaline-earth oxides 579:632 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 0 3 theme non-carbohydrate 81:96 arg1 components 98:107 non-carbohydrate components 81:107 non-carbohydrate components of bagasse and straw 81:128 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 4 4 theme alkaline-earth 612:625 arg1 oxides 627:632 alkaline and alkaline-earth oxides 599:632 alkaline and alkaline-earth oxides 599:632 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 1 5 theme chemical 236:243 arg1 composition 245:255 their chemical composition 230:255 their chemical composition 230:255 Two fractions of sugarcane, namely bagasse and straw (or trash), were characterized in relation to their chemical composition. 25263869 0 6 theme bagasse 112:118 arg1 components 98:107 non-carbohydrate components 81:107 non-carbohydrate components of bagasse and straw 81:128 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 0 6 theme bagasse 112:118 arg1 composition 49:59 comparative composition 37:59 comparative composition of carbohydrate 37:75 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 2 7 contain had 386:388 arg1 straw 380:384 straw 380:384 straw 380:384 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 7 contain had 386:388 arg2 components 434:443 these same components 423:443 these same components 423:443 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 7 contain had 386:388 arg2 % 418:418 33.77, 27.38, 21.28 and 6.23% 390:418 33.77, 27.38, 21.28 and 6.23% of these same components 390:443 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 3 8 from high 473:476 arg1 fractions 499:507 both cane biomass fractions 481:507 both cane biomass fractions 481:507 Ash content was relatively high in both cane biomass fractions. 25263869 5 9 theme polysaccharide 660:673 arg1 compositions 684:695 the polysaccharide chemical compositions 656:695 the polysaccharide chemical compositions of these lignocellulosic materials 656:730 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 6 10 theme enzyme 1054:1059 arg1 cocktails 1061:1069 more efficient enzyme cocktails 1039:1069 more efficient enzyme cocktails 1039:1069 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 11 theme substrate 1075:1083 arg1 saccharification 1085:1100 substrate saccharification 1075:1100 substrate saccharification 1075:1100 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 2 12 theme components 434:443 arg1 components 434:443 these same components 423:443 these same components 423:443 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 12 theme components 434:443 arg1 % 418:418 33.77, 27.38, 21.28 and 6.23% 390:418 33.77, 27.38, 21.28 and 6.23% of these same components 390:443 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 4 13 theme oxides 556:561 arg1 levels 532:537 higher levels 525:537 higher levels of contaminating oxides 525:561 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 2 14 theme % 357:357 arg1 ash 322:324 ash 322:324 ash 322:324 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 14 theme % 357:357 arg1 lignin 311:316 lignin 311:316 lignin 311:316 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 14 theme % 357:357 arg1 glucans 286:292 glucans 286:292 glucans 286:292 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 2 15 theme same 429:432 arg1 components 434:443 these same components 423:443 these same components 423:443 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 5 16 theme fermentable 765:775 arg1 sugars 777:782 fermentable sugars 765:782 fermentable sugars 765:782 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 6 17 theme efficient 1044:1052 arg1 cocktails 1061:1069 more efficient enzyme cocktails 1039:1069 more efficient enzyme cocktails 1039:1069 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 0 18 theme straw 124:128 arg1 components 98:107 non-carbohydrate components 81:107 non-carbohydrate components of bagasse and straw 81:128 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 0 18 theme straw 124:128 arg1 composition 49:59 comparative composition 37:59 comparative composition of carbohydrate 37:75 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 5 19 theme materials 722:730 arg1 compositions 684:695 the polysaccharide chemical compositions 656:695 the polysaccharide chemical compositions of these lignocellulosic materials 656:730 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 3 20 theme cane 486:489 arg1 fractions 499:507 both cane biomass fractions 481:507 both cane biomass fractions 481:507 Ash content was relatively high in both cane biomass fractions. 25263869 0 21 theme Sugarcane 0:8 arg1 biomass 10:16 Sugarcane biomass 0:16 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 0:129 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 3 22 theme biomass 491:497 arg1 fractions 499:507 both cane biomass fractions 481:507 both cane biomass fractions 481:507 Ash content was relatively high in both cane biomass fractions. 25263869 3 23 from fractions 499:507 arg1 high 473:476 high 473:476 high 473:476 Ash content was relatively high in both cane biomass fractions. 25263869 5 24 theme sugars 777:782 arg1 amounts 754:760 similar amounts 746:760 similar amounts of fermentable sugars 746:782 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 5 24 theme sugars 777:782 arg1 sugars 777:782 fermentable sugars 765:782 fermentable sugars 765:782 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 6 25 theme good 1008:1011 arg1 opportunity 1013:1023 a good opportunity 1006:1023 a good opportunity for designing more efficient enzyme cocktails for substrate saccharification 1006:1100 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 26 theme hemicelluloses 933:946 arg1 properties 891:900 the chemical properties 878:900 the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose 878:970 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 2 27 theme glucans 286:292 arg1 values 276:281 values 276:281 values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53% 276:357 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 1 28 theme sugarcane 148:156 arg1 fractions 135:143 Two fractions 131:143 Two fractions of sugarcane 131:156 Two fractions of sugarcane, namely bagasse and straw (or trash), were characterized in relation to their chemical composition. 25263869 1 28 theme sugarcane 148:156 arg1 straw 178:182 straw 178:182 straw 178:182 Two fractions of sugarcane, namely bagasse and straw (or trash), were characterized in relation to their chemical composition. 25263869 1 28 theme sugarcane 148:156 arg1 bagasse 166:172 bagasse 166:172 bagasse 166:172 Two fractions of sugarcane, namely bagasse and straw (or trash), were characterized in relation to their chemical composition. 25263869 0 29 theme comparative 37:47 arg1 composition 49:59 comparative composition 37:59 comparative composition of carbohydrate 37:75 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 4 30 contain had 575:577 arg1 straw 569:573 straw 569:573 straw 569:573 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 4 30 contain had 575:577 arg2 content 588:594 a higher content 579:594 a higher content of alkaline and alkaline-earth oxides 579:632 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 4 31 theme higher 581:586 arg1 content 588:594 a higher content 579:594 a higher content of alkaline and alkaline-earth oxides 579:632 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 6 32 theme holocellulose 918:930 arg1 properties 891:900 the chemical properties 878:900 the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose 878:970 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 33 theme α-cellulose 960:970 arg1 properties 891:900 the chemical properties 878:900 the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose 878:970 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 5 34 theme similar 746:752 arg1 amounts 754:760 similar amounts 746:760 similar amounts of fermentable sugars 746:782 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 5 34 theme similar 746:752 arg1 sugars 777:782 fermentable sugars 765:782 fermentable sugars 765:782 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 5 35 theme enzymatic 842:850 arg1 hydrolysis 852:861 enzymatic hydrolysis 842:861 enzymatic hydrolysis 842:861 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 6 36 theme cane 905:908 arg1 holocellulose 918:930 cane biomass holocellulose 905:930 cane biomass holocellulose 905:930 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 37 dep hemicelluloses 933:946 arg1 hemicelluloses 933:946 hemicelluloses A and B 933:954 hemicelluloses A and B 933:954 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 37 dep hemicelluloses 933:946 arg1 B 954:954 B 954:954 B 954:954 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 6 37 dep hemicelluloses 933:946 arg1 A 948:948 A 948:948 A 948:948 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25263869 5 38 theme chemical 675:682 arg1 compositions 684:695 the polysaccharide chemical compositions 656:695 the polysaccharide chemical compositions of these lignocellulosic materials 656:730 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 4 39 theme alkaline 599:606 arg1 oxides 627:632 alkaline and alkaline-earth oxides 599:632 alkaline and alkaline-earth oxides 599:632 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 4 40 theme higher 525:530 arg1 levels 532:537 higher levels 525:537 higher levels of contaminating oxides 525:561 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 0 41 dep biomass 10:16 arg1 components 98:107 non-carbohydrate components 81:107 non-carbohydrate components of bagasse and straw 81:128 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 0 41 dep biomass 10:16 arg1 composition 49:59 comparative composition 37:59 comparative composition of carbohydrate 37:75 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 2 42 dep presented 266:274 arg1 while 374:378 while 374:378 while 374:378 Bagasse presented values of glucans, hemicelluloses, lignin and ash of 37.74, 27.23, 20.57 and 6.53%, respectively, while straw had 33.77, 27.38, 21.28 and 6.23% of these same components. 25263869 5 43 theme optimal 817:823 arg1 pretreatment 825:836 optimal pretreatment 817:836 optimal pretreatment 817:836 A comparison between the polysaccharide chemical compositions of these lignocellulosic materials suggests that similar amounts of fermentable sugars are expected to arise from their optimal pretreatment and enzymatic hydrolysis. 25263869 4 44 theme contaminating 542:554 arg1 oxides 556:561 contaminating oxides 542:561 contaminating oxides 542:561 Bagasse showed higher levels of contaminating oxides while straw had a higher content of alkaline and alkaline-earth oxides. 25263869 0 45 theme carbohydrate 64:75 arg1 components 98:107 non-carbohydrate components 81:107 non-carbohydrate components of bagasse and straw 81:128 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 0 45 theme carbohydrate 64:75 arg1 composition 49:59 comparative composition 37:59 comparative composition of carbohydrate 37:75 Sugarcane biomass for biorefineries: comparative composition of carbohydrate and non-carbohydrate components of bagasse and straw. 25263869 3 46 theme Ash 446:448 arg1 content 450:456 Ash content 446:456 Ash content 446:456 Ash content was relatively high in both cane biomass fractions. 25263869 6 47 theme biomass 910:916 arg1 holocellulose 918:930 cane biomass holocellulose 905:930 cane biomass holocellulose 905:930 Details about the chemical properties of cane biomass holocellulose, hemicelluloses A and B and α-cellulose are provided, and these may offer a good opportunity for designing more efficient enzyme cocktails for substrate saccharification. 25030939 1 0 theme diabetes 326:333 arg1 mellitus 335:342 type 1 and type 2 diabetes mellitus 308:342 mellitus 335:342 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 2 1 gly N-glycosylation 402:416 arg1 pramlintide 421:431 pramlintide 421:431 pramlintide 421:431 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 4 2 used used 970:973 arg2 N-glycosylation 947:961 N-glycosylation 947:961 N-glycosylation 947:961 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 2 used used 970:973 arg2 tool 980:983 a tool 978:983 a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity 978:1126 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 3 theme synthetic 502:510 arg1 route 512:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 4 theme pharmacokinetic 1001:1015 arg1 properties 1017:1026 the pharmacokinetic properties 997:1026 the pharmacokinetic properties of pramlintide 997:1041 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 0 5 theme pramlintide 78:88 arg1 analogues 65:73 glycosylated analogues 52:73 glycosylated analogues of pramlintide 52:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 6 theme core 694:697 arg1 GlcNAc 730:735 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 7 gly glycosylated 863:874 arg1 versions 876:883 glycosylated versions 863:883 glycosylated versions of pramlintide 863:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 8 theme pramlintide 1031:1041 arg1 properties 1017:1026 the pharmacokinetic properties 997:1026 the pharmacokinetic properties of pramlintide 997:1041 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 9 theme receptor 912:919 arg1 agonists 921:928 potent receptor agonists 905:928 potent receptor agonists 905:928 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 9 theme receptor 912:919 arg1 majority 851:858 The majority 847:858 The majority of glycosylated versions of pramlintide 847:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 10 theme N-glycan 699:706 arg1 GlcNAc 730:735 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 2 11 contain has 433:435 arg1 N-glycosylation 402:416 N-glycosylation 402:416 N-glycosylation of pramlintide 402:431 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 2 11 contain has 433:435 arg2 effect 390:395 the effect 386:395 the effect that N-glycosylation of pramlintide has on activation of amylin receptors 386:469 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 3 12 theme asparagine 826:835 arg1 residues 837:844 six asparagine residues 822:844 its six asparagine residues 818:844 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 0 13 theme structure-activity 91:108 arg1 relationships 110:122 structure-activity relationships 91:122 structure-activity relationships for amylin receptor agonism 91:150 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 14 theme solid 546:550 arg1 synthesis 566:574 solid phase peptide synthesis 546:574 solid phase peptide synthesis 546:574 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 0 15 theme amylin 128:133 arg1 agonism 144:150 amylin receptor agonism 128:150 amylin receptor agonism 128:150 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 0 16 dep relationships 110:122 arg1 synthesis 26:34 Convergent chemoenzymatic synthesis 0:34 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide 0:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 4 17 theme improved 1058:1065 arg1 agents 1079:1084 improved therapeutic agents 1058:1084 improved therapeutic agents 1058:1084 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 18 gly N-glycosylated 628:641 arg1 variants 643:650 N-glycosylated variants 628:650 N-glycosylated variants of pramlintide bearing either GlcNAc 628:687 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 1 19 theme synthetic 178:186 arg1 analogue 188:195 a synthetic analogue 176:195 a synthetic analogue of the naturally occurring pancreatic hormone amylin 176:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 1 19 theme synthetic 178:186 arg1 Pramlintide 153:163 Pramlintide 153:163 Pramlintide (Symlin®) 153:173 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 0 20 theme chemoenzymatic 11:24 arg1 synthesis 26:34 Convergent chemoenzymatic synthesis 0:34 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide 0:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 21 theme GlcNAc 730:735 arg1 library 617:623 a library 615:623 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 22 theme synthesis 566:574 arg1 combination 531:541 a combination 529:541 a combination of solid phase peptide synthesis 529:574 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 22 theme synthesis 566:574 arg1 glycosylation 590:602 enzymatic glycosylation 580:602 enzymatic glycosylation 580:602 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 23 theme diabetes 1107:1114 arg1 treatment 1094:1102 the treatment 1090:1102 the treatment of diabetes and obesity 1090:1126 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 0 24 theme Convergent 0:9 arg1 synthesis 26:34 Convergent chemoenzymatic synthesis 0:34 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide 0:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 2 25 theme systematic 364:373 arg1 study 375:379 a systematic study 362:379 a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors 362:469 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 1 26 theme adjunctive 285:294 arg1 therapy 296:302 adjunctive therapy 285:302 adjunctive therapy for type 1 and type 2 diabetes mellitus 285:342 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 4 27 dep and 1043:1045 arg1 so 1047:1048 so 1047:1048 so 1047:1048 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 28 theme phase 552:556 arg1 synthesis 566:574 solid phase peptide synthesis 546:574 solid phase peptide synthesis 546:574 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 29 theme therapeutic 1067:1077 arg1 agents 1079:1084 improved therapeutic agents 1058:1084 improved therapeutic agents 1058:1084 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 2 30 theme pramlintide 421:431 arg1 N-glycosylation 402:416 N-glycosylation 402:416 N-glycosylation of pramlintide 402:431 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 0 31 theme receptor 135:142 arg1 agonism 144:150 amylin receptor agonism 128:150 amylin receptor agonism 128:150 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 32 theme pramlintide 655:665 arg1 bearing 667:673 pramlintide bearing 655:673 pramlintide bearing 655:673 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 33 theme peptide 558:564 arg1 synthesis 566:574 solid phase peptide synthesis 546:574 solid phase peptide synthesis 546:574 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 2 34 theme amylin 454:459 arg1 receptors 461:469 amylin receptors 454:469 amylin receptors 454:469 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 3 35 dep library 617:623 arg1 GlcNAc 797:802 (NeuAcGalGlcNAcMan)2Man(GlcNAc)2 773:804 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 0 36 theme library 41:47 arg1 synthesis 26:34 Convergent chemoenzymatic synthesis 0:34 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide 0:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 37 theme bearing 667:673 arg1 glycan 765:770 a complex biantennary glycan 743:770 a complex biantennary glycan 743:770 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 37 theme bearing 667:673 arg1 variants 643:650 N-glycosylated variants 628:650 N-glycosylated variants of pramlintide bearing either GlcNAc 628:687 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 37 theme bearing 667:673 arg1 GlcNAc 730:735 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 38 theme potent 905:910 arg1 agonists 921:928 potent receptor agonists 905:928 potent receptor agonists 905:928 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 38 theme potent 905:910 arg1 majority 851:858 The majority 847:858 The majority of glycosylated versions of pramlintide 847:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 39 gly glycosylation 590:602 arg1 synthesis 566:574 solid phase peptide synthesis 546:574 solid phase peptide synthesis 546:574 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 1 40 theme occurring 214:222 arg1 amylin 243:248 the naturally occurring pancreatic hormone amylin 200:248 the naturally occurring pancreatic hormone amylin 200:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 2 41 theme receptors 461:469 arg1 activation 440:449 activation 440:449 activation of amylin receptors 440:469 Herein we report a systematic study into the effect that N-glycosylation of pramlintide has on activation of amylin receptors. 25030939 4 42 theme pramlintide 888:898 arg1 versions 876:883 glycosylated versions 863:883 glycosylated versions of pramlintide 863:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 1 43 used used 264:267 arg2 Symlin® 166:172 Symlin® 166:172 Symlin® 166:172 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 1 43 used used 264:267 arg2 Pramlintide 153:163 Pramlintide 153:163 Pramlintide (Symlin®) 153:173 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 1 43 used used 264:267 arg2 analogue 188:195 a synthetic analogue 176:195 a synthetic analogue of the naturally occurring pancreatic hormone amylin 176:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 1 44 theme pancreatic 224:233 arg1 amylin 243:248 the naturally occurring pancreatic hormone amylin 200:248 the naturally occurring pancreatic hormone amylin 200:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 0 45 theme glycosylated 52:63 arg1 analogues 65:73 glycosylated analogues 52:73 glycosylated analogues of pramlintide 52:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 46 theme complex 745:751 arg1 glycan 765:770 a complex biantennary glycan 743:770 a complex biantennary glycan 743:770 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 47 theme enzymatic 580:588 arg1 glycosylation 590:602 enzymatic glycosylation 580:602 enzymatic glycosylation 580:602 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 48 theme efficient 481:489 arg1 route 512:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 49 theme biantennary 753:763 arg1 glycan 765:770 a complex biantennary glycan 743:770 a complex biantennary glycan 743:770 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 50 theme variants 643:650 arg1 library 617:623 a library 615:623 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 1 51 theme type 319:322 arg1 mellitus 335:342 type 1 and type 2 diabetes mellitus 308:342 mellitus 335:342 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 4 52 theme glycosylated 863:874 arg1 versions 876:883 glycosylated versions 863:883 glycosylated versions of pramlintide 863:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 1 53 theme hormone 235:241 arg1 amylin 243:248 the naturally occurring pancreatic hormone amylin 200:248 the naturally occurring pancreatic hormone amylin 200:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 3 54 theme glycan 765:770 arg1 library 617:623 a library 615:623 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 55 theme N-glycosylated 628:641 arg1 variants 643:650 N-glycosylated variants 628:650 N-glycosylated variants of pramlintide bearing either GlcNAc 628:687 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 4 56 theme versions 876:883 arg1 agonists 921:928 potent receptor agonists 905:928 potent receptor agonists 905:928 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 4 56 theme versions 876:883 arg1 majority 851:858 The majority 847:858 The majority of glycosylated versions of pramlintide 847:898 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 1 57 theme amylin 243:248 arg1 analogue 188:195 a synthetic analogue 176:195 a synthetic analogue of the naturally occurring pancreatic hormone amylin 176:248 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 1 57 theme amylin 243:248 arg1 Pramlintide 153:163 Pramlintide 153:163 Pramlintide (Symlin®) 153:173 Pramlintide (Symlin®), a synthetic analogue of the naturally occurring pancreatic hormone amylin, is currently used with insulin in adjunctive therapy for type 1 and type 2 diabetes mellitus. 25030939 4 58 theme obesity 1120:1126 arg1 treatment 1094:1102 the treatment 1090:1102 the treatment of diabetes and obesity 1090:1126 The majority of glycosylated versions of pramlintide were potent receptor agonists, suggesting that N-glycosylation may be used as a tool to optimise the pharmacokinetic properties of pramlintide and so deliver improved therapeutic agents for the treatment of diabetes and obesity. 25030939 3 59 theme NeuAcGalGlcNAcMan 774:790 arg1 GlcNAc 797:802 (NeuAcGalGlcNAcMan)2Man(GlcNAc)2 773:804 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 0 60 theme analogues 65:73 arg1 library 41:47 a library 39:47 a library of glycosylated analogues of pramlintide 39:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 61 theme pentasaccharide 708:722 arg1 GlcNAc 730:735 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 0 62 gly glycosylated 52:63 arg1 analogues 65:73 glycosylated analogues 52:73 glycosylated analogues of pramlintide 52:88 Convergent chemoenzymatic synthesis of a library of glycosylated analogues of pramlintide: structure-activity relationships for amylin receptor agonism. 25030939 3 63 theme [Man3 724:728 arg1 GlcNAc 730:735 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 the core N-glycan pentasaccharide [Man3(GlcNAc)2] 690:738 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 64 theme 2Man 792:795 arg1 GlcNAc 797:802 (NeuAcGalGlcNAcMan)2Man(GlcNAc)2 773:804 a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] 615:805 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 65 theme convergent 491:500 arg1 route 512:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route 472:516 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25030939 3 66 dep bearing 667:673 arg1 either 675:680 either 675:680 either 675:680 A highly efficient convergent synthetic route, involving a combination of solid phase peptide synthesis and enzymatic glycosylation, delivered a library of N-glycosylated variants of pramlintide bearing either GlcNAc, the core N-glycan pentasaccharide [Man3(GlcNAc)2] or a complex biantennary glycan [(NeuAcGalGlcNAcMan)2Man(GlcNAc)2] at each of its six asparagine residues. 25209049 7 0 theme lectin-exosome 1099:1112 arg1 assay 1122:1126 the lectin-exosome binding assay 1095:1126 the lectin-exosome binding assay 1095:1126 The uEVs were subsequently immobilized on solid phase and probed with labeled plant lectins using the lectin-exosome binding assay. 25209049 5 1 theme differential 868:879 arg1 centrifugation 881:894 differential centrifugation 868:894 differential centrifugation 868:894 Salt precipitation was employed to remove THP under conditions originally described for its separation from urine, followed by differential centrifugation. 25209049 9 2 theme lectin 1304:1309 arg1 reference 1375:1383 a reference 1373:1383 a reference for assessing the composition of their surface glycans in different physiological and pathological conditions 1373:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 2 theme lectin 1304:1309 arg1 pattern 1319:1325 The basic lectin binding pattern 1294:1325 The basic lectin binding pattern of uEVs obtained by our method 1294:1356 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 5 3 theme Salt 741:744 arg1 precipitation 746:758 Salt precipitation 741:758 Salt precipitation 741:758 Salt precipitation was employed to remove THP under conditions originally described for its separation from urine, followed by differential centrifugation. 25209049 7 4 theme binding 1114:1120 arg1 assay 1122:1126 the lectin-exosome binding assay 1095:1126 the lectin-exosome binding assay 1095:1126 The uEVs were subsequently immobilized on solid phase and probed with labeled plant lectins using the lectin-exosome binding assay. 25209049 3 5 gly glycosylated 496:507 arg1 THP 481:483 THP 481:483 THP 481:483 This may make glycan analysis of uEVs difficult since THP is heavily glycosylated. 25209049 0 6 from Isolation 0:8 arg1 Horsfall 55:62 Tamm- Horsfall 49:62 Tamm- Horsfall 49:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 9 7 theme surface 1424:1430 arg1 glycans 1432:1438 their surface glycans 1418:1438 their surface glycans 1418:1438 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 2 8 theme urinary 289:295 arg1 uEVs 302:305 uEVs 302:305 uEVs 302:305 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 2 8 theme urinary 289:295 arg1 EVs 297:299 urinary EVs 289:299 urinary EVs (uEVs) 289:306 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 9 9 theme basic 1298:1302 arg1 reference 1375:1383 a reference 1373:1383 a reference for assessing the composition of their surface glycans in different physiological and pathological conditions 1373:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 9 theme basic 1298:1302 arg1 pattern 1319:1325 The basic lectin binding pattern 1294:1325 The basic lectin binding pattern of uEVs obtained by our method 1294:1356 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 0 10 from Horsfall 55:62 arg1 Isolation 0:8 Isolation 0:8 Isolation of urinary extracellular vesicles from Tamm- Horsfall 0:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 0 10 from Horsfall 55:62 arg1 vesicles 35:42 urinary extracellular vesicles 13:42 urinary extracellular vesicles from Tamm- Horsfall 13:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 4 11 theme binding 726:732 arg1 assay 734:738 a lectin-exosome binding assay 709:738 a lectin-exosome binding assay 709:738 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 8 12 theme isolated 1159:1166 arg1 uEVs 1168:1171 the isolated uEVs 1155:1171 the isolated uEVs 1155:1171 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 9 13 theme binding 1311:1317 arg1 reference 1375:1383 a reference 1373:1383 a reference for assessing the composition of their surface glycans in different physiological and pathological conditions 1373:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 13 theme binding 1311:1317 arg1 pattern 1319:1325 The basic lectin binding pattern 1294:1325 The basic lectin binding pattern of uEVs obtained by our method 1294:1356 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 7 14 theme plant 1075:1079 arg1 lectins 1081:1087 labeled plant lectins 1067:1087 labeled plant lectins 1067:1087 The uEVs were subsequently immobilized on solid phase and probed with labeled plant lectins using the lectin-exosome binding assay. 25209049 3 15 theme glycan 441:446 arg1 analysis 448:455 glycan analysis 441:455 glycan analysis of uEVs 441:463 This may make glycan analysis of uEVs difficult since THP is heavily glycosylated. 25209049 8 16 dep selective 1252:1260 arg1 carbohydrate-dependent 1263:1284 carbohydrate-dependent 1263:1284 carbohydrate-dependent 1263:1284 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 9 17 theme glycans 1432:1438 arg1 composition 1403:1413 the composition 1399:1413 the composition of their surface glycans in different physiological and pathological conditions 1399:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 8 18 theme plant 1233:1237 arg1 lectins 1239:1245 labeled plant lectins 1225:1245 labeled plant lectins 1225:1245 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 1 19 theme available 182:190 arg1 source 192:197 a readily available source 172:197 a readily available source of relatively large quantities of extracellular vesicles (EVs) 172:260 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 1 19 theme available 182:190 arg1 Urine 163:167 Urine 163:167 Urine 163:167 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 9 20 used used 1365:1368 arg2 pattern 1319:1325 The basic lectin binding pattern 1294:1325 The basic lectin binding pattern of uEVs obtained by our method 1294:1356 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 20 used used 1365:1368 arg2 reference 1375:1383 a reference 1373:1383 a reference for assessing the composition of their surface glycans in different physiological and pathological conditions 1373:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 4 21 theme isolation 641:649 arg1 procedure 651:659 the uEV isolation procedure 633:659 the uEV isolation procedure 633:659 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 7 22 theme solid 1039:1043 arg1 phase 1045:1049 solid phase 1039:1049 solid phase 1039:1049 The uEVs were subsequently immobilized on solid phase and probed with labeled plant lectins using the lectin-exosome binding assay. 25209049 9 23 theme uEVs 1330:1333 arg1 reference 1375:1383 a reference 1373:1383 a reference for assessing the composition of their surface glycans in different physiological and pathological conditions 1373:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 23 theme uEVs 1330:1333 arg1 pattern 1319:1325 The basic lectin binding pattern 1294:1325 The basic lectin binding pattern of uEVs obtained by our method 1294:1356 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 7 24 theme labeled 1067:1073 arg1 lectins 1081:1087 labeled plant lectins 1067:1087 labeled plant lectins 1067:1087 The uEVs were subsequently immobilized on solid phase and probed with labeled plant lectins using the lectin-exosome binding assay. 25209049 4 25 theme uEV 637:639 arg1 procedure 651:659 the uEV isolation procedure 633:659 the uEV isolation procedure 633:659 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 0 26 theme extracellular 21:33 arg1 vesicles 35:42 urinary extracellular vesicles 13:42 urinary extracellular vesicles from Tamm- Horsfall 13:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 4 27 theme glycosylation 524:536 arg1 analysis 538:545 glycosylation analysis 524:545 glycosylation analysis 524:545 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 9 28 from composition 1403:1413 arg1 conditions 1484:1493 different physiological and pathological conditions 1443:1493 different physiological and pathological conditions 1443:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 4 29 theme lectin-exosome 711:724 arg1 assay 734:738 a lectin-exosome binding assay 709:738 a lectin-exosome binding assay 709:738 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 8 30 theme selective 1252:1260 arg1 manner 1286:1291 a selective, carbohydrate-dependent manner 1250:1291 a selective, carbohydrate-dependent manner 1250:1291 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 0 31 theme urinary 13:19 arg1 vesicles 35:42 urinary extracellular vesicles 13:42 urinary extracellular vesicles from Tamm- Horsfall 13:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 4 32 theme non-uEV 587:593 arg1 glycans 595:601 non-uEV glycans 587:601 non-uEV glycans 587:601 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 2 33 theme EVs 297:299 arg1 isolation 276:284 the isolation 272:284 the isolation of urinary EVs (uEVs) 272:306 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 1 34 theme large 213:217 arg1 quantities 219:228 relatively large quantities 202:228 relatively large quantities of extracellular vesicles (EVs) 202:260 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 0 35 theme assay 156:160 arg1 development 116:126 the development 112:126 the development of a lectin-exosome-binding assay 112:160 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 1 36 theme quantities 219:228 arg1 source 192:197 a readily available source 172:197 a readily available source of relatively large quantities of extracellular vesicles (EVs) 172:260 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 1 36 theme quantities 219:228 arg1 Urine 163:167 Urine 163:167 Urine 163:167 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 0 37 theme vesicles 35:42 arg1 Isolation 0:8 Isolation 0:8 Isolation of urinary extracellular vesicles from Tamm- Horsfall 0:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 0 38 theme lectin-exosome-binding 133:154 arg1 assay 156:160 a lectin-exosome-binding assay 131:160 a lectin-exosome-binding assay 131:160 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 0 39 from urine 81:85 arg1 development 116:126 the development 112:126 the development of a lectin-exosome-binding assay 112:160 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 8 40 theme structural 1187:1196 arg1 integrity 1198:1206 structural integrity 1187:1206 structural integrity 1187:1206 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 6 41 theme uEVs 925:928 arg1 quality 901:907 The quality 897:907 The quality of the isolated uEVs 897:928 The quality of the isolated uEVs was examined by electron microscopy, SDS-PAGE, and immunoblotting. 25209049 6 42 theme electron 946:953 arg1 microscopy 955:964 electron microscopy 946:964 electron microscopy 946:964 The quality of the isolated uEVs was examined by electron microscopy, SDS-PAGE, and immunoblotting. 25209049 3 43 theme uEVs 460:463 arg1 analysis 448:455 glycan analysis 441:455 glycan analysis of uEVs 441:463 This may make glycan analysis of uEVs difficult since THP is heavily glycosylated. 25209049 0 44 theme Tamm- 49:53 arg1 Horsfall 55:62 Tamm- Horsfall 49:62 Tamm- Horsfall 49:62 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 6 45 theme isolated 916:923 arg1 uEVs 925:928 the isolated uEVs 912:928 the isolated uEVs 912:928 The quality of the isolated uEVs was examined by electron microscopy, SDS-PAGE, and immunoblotting. 25209049 4 46 theme isolated 673:680 arg1 uEVs 682:685 the isolated uEVs 669:685 the isolated uEVs in the development of a lectin-exosome binding assay 669:738 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 1 47 theme extracellular 233:245 arg1 EVs 257:259 EVs 257:259 EVs 257:259 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 1 47 theme extracellular 233:245 arg1 vesicles 247:254 extracellular vesicles 233:254 extracellular vesicles (EVs) 233:260 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 4 48 theme assay 734:738 arg1 development 694:704 the development 690:704 the development of a lectin-exosome binding assay 690:738 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 5 49 from urine 849:853 arg1 separation 833:842 its separation 829:842 its separation 829:842 Salt precipitation was employed to remove THP under conditions originally described for its separation from urine, followed by differential centrifugation. 25209049 4 50 theme procedure 651:659 arg1 modification 617:628 a modification 615:628 a modification of the uEV isolation procedure 615:659 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 4 51 from uEVs 682:685 arg1 development 694:704 the development 690:704 the development of a lectin-exosome binding assay 690:738 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25209049 1 52 theme vesicles 247:254 arg1 quantities 219:228 relatively large quantities 202:228 relatively large quantities of extracellular vesicles (EVs) 202:260 Urine is a readily available source of relatively large quantities of extracellular vesicles (EVs). 25209049 0 53 from application 97:107 arg1 development 116:126 the development 112:126 the development of a lectin-exosome-binding assay 112:160 Isolation of urinary extracellular vesicles from Tamm- Horsfall protein-depleted urine and their application in the development of a lectin-exosome-binding assay. 25209049 9 54 dep different 1443:1451 arg1 physiological 1453:1465 physiological 1453:1465 physiological 1453:1465 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 54 dep different 1443:1451 arg1 pathological 1471:1482 pathological 1471:1482 pathological 1471:1482 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 9 55 theme different 1443:1451 arg1 conditions 1484:1493 different physiological and pathological conditions 1443:1493 different physiological and pathological conditions 1443:1493 The basic lectin binding pattern of uEVs obtained by our method can be used as a reference for assessing the composition of their surface glycans in different physiological and pathological conditions. 25209049 2 56 theme protein 356:362 arg1 presence 330:337 the presence 326:337 the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant 326:424 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 8 57 theme labeled 1225:1231 arg1 lectins 1239:1245 labeled plant lectins 1225:1245 labeled plant lectins 1225:1245 Our results indicate that the isolated uEVs had preserved structural integrity and reacted with labeled plant lectins in a selective, carbohydrate-dependent manner. 25209049 2 58 theme Tamm-Horsfall 342:354 arg1 THP 365:367 THP 365:367 THP 365:367 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 2 58 theme Tamm-Horsfall 342:354 arg1 protein 356:362 Tamm-Horsfall protein 342:362 Tamm-Horsfall protein (THP) 342:368 However, the isolation of urinary EVs (uEVs) is complicated by the presence of Tamm-Horsfall protein (THP), which polymerizes and co-precipitates as a contaminant. 25209049 4 59 theme glycans 595:601 arg1 elimination 572:582 elimination 572:582 elimination of non-uEV glycans 572:601 To facilitate glycosylation analysis and address the need for elimination of non-uEV glycans, we present a modification of the uEV isolation procedure and use the isolated uEVs in the development of a lectin-exosome binding assay. 25326312 10 0 theme impaired 2343:2350 arg1 movement 2364:2371 impaired coordinated movement 2343:2371 impaired coordinated movement 2343:2371 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 4 1 link O-linked 1015:1022 arg1 glycans 1024:1030 O-linked glycans 1015:1030 O-linked glycans 1015:1030 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 3 2 theme cognitive 784:792 arg1 impairments 794:804 striking neurodevelopmental, motor and cognitive impairments 745:804 striking neurodevelopmental, motor and cognitive impairments 745:804 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 9 3 from changes 2047:2053 arg1 co-receptor 2094:2104 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 9 3 from changes 2047:2053 arg1 ligand 2114:2119 Wnt ligand 2110:2119 Wnt ligand 2110:2119 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 9 4 theme Wnt 2110:2112 arg1 ligand 2114:2119 Wnt ligand 2110:2119 Wnt ligand 2110:2119 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 8 5 theme moieties 1809:1816 arg1 depletion 1717:1725 depletion 1717:1725 depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1717:1816 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 8 6 theme sugarless 1878:1886 arg1 overexpression 1888:1901 sugarless overexpression 1878:1901 sugarless overexpression 1878:1901 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 1 7 theme galactose-1-phosphate 291:311 arg1 conversion 277:286 conversion 277:286 conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis 277:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 4 8 theme glycosylation 963:975 arg1 CG 907:908 CG 907:908 CG 907:908 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 4 8 theme glycosylation 963:975 arg1 disorder 951:958 a congenital disorder 938:958 a congenital disorder of glycosylation (CDG) 938:981 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 7 9 theme cellular 1561:1568 arg1 defects 1570:1576 behavioral and cellular defects 1546:1576 behavioral and cellular defects 1546:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 10 theme behavioral 1546:1555 arg1 defects 1570:1576 behavioral and cellular defects 1546:1576 behavioral and cellular defects 1546:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 3 11 theme acute 663:667 arg1 symptoms 698:705 acute, potentially lethal neonatal symptoms 663:705 acute, potentially lethal neonatal symptoms 663:705 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 6 12 theme structural 1266:1275 arg1 overelaboration 1277:1291 structural overelaboration 1266:1291 structural overelaboration 1266:1291 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 1 13 theme galactose-1-phosphate 213:233 arg1 GALT 254:257 GALT 254:257 GALT 254:257 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 13 theme galactose-1-phosphate 213:233 arg1 uridyltransferase 235:251 galactose-1-phosphate uridyltransferase 213:251 galactose-1-phosphate uridyltransferase (GALT) 213:258 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 2 14 theme extracellular 626:638 arg1 spaces 640:645 extracellular spaces 626:645 extracellular spaces 626:645 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 10 15 theme neurological 2381:2392 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 1 16 theme synthesis 471:479 arg1 upstream 405:412 upstream 405:412 upstream 405:412 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 3 17 theme neonatal 689:696 arg1 symptoms 698:705 acute, potentially lethal neonatal symptoms 663:705 acute, potentially lethal neonatal symptoms 663:705 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 8 18 theme extracellular 1596:1608 arg1 synaptomatrix 1610:1622 the NMJ extracellular synaptomatrix 1588:1622 the NMJ extracellular synaptomatrix with a broad panel of lectin probes 1588:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 7 19 theme critical 1498:1505 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 19 theme critical 1498:1505 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 5 20 theme disease 1167:1173 arg1 model 1175:1179 a GALT-deficient Drosophila disease model 1139:1179 a GALT-deficient Drosophila disease model 1139:1179 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 1 21 dep galactose-1-phosphate 291:311 arg1 -glucose 342:349 -glucose 342:349 -glucose 342:349 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 9 22 theme sugarless 2178:2186 arg1 overexpression 2188:2201 sugarless overexpression 2178:2201 sugarless overexpression 2178:2201 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 4 23 theme neurological 837:848 arg1 symptoms 850:857 neurological symptoms 837:857 neurological symptoms 837:857 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 8 24 theme broad 1631:1635 arg1 panel 1637:1641 a broad panel 1629:1641 a broad panel of lectin probes 1629:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 10 25 theme synaptomatrix 2225:2237 arg1 losses 2253:2258 synaptomatrix glycosylation losses 2225:2258 synaptomatrix glycosylation losses 2225:2258 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 7 26 theme galactose 1370:1378 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 26 theme galactose 1370:1378 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 4 27 from defects 992:998 arg1 N- 1008:1009 N- 1008:1009 N- 1008:1009 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 4 27 from defects 992:998 arg1 glycans 1024:1030 O-linked glycans 1015:1030 O-linked glycans 1015:1030 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 2 28 theme glycolipids 572:582 arg1 synthesis 541:549 the synthesis 537:549 the synthesis of glycoproteins and glycolipids 537:582 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 4 29 theme Previous 807:814 arg1 studies 816:822 Previous studies 807:822 Previous studies 807:822 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 10 30 theme signaling 2284:2292 arg1 components 2302:2311 altered trans-synaptic signaling pathway components 2261:2311 altered trans-synaptic signaling pathway components 2261:2311 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 0 31 theme classic 93:99 arg1 model 122:126 a Drosophila classic galactosemia disease model 80:126 a Drosophila classic galactosemia disease model 80:126 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 1 32 theme recessive 171:179 arg1 disorder 181:188 an autosomal recessive disorder 158:188 an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis 158:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 32 theme recessive 171:179 arg1 galactosemia 137:148 Classic galactosemia 129:148 Classic galactosemia (CG) 129:153 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 9 33 theme synaptomatrix 1970:1982 arg1 environment 1997:2007 this synaptomatrix carbohydrate environment 1965:2007 this synaptomatrix carbohydrate environment 1965:2007 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 8 34 theme probes 1653:1658 arg1 panel 1637:1641 a broad panel 1629:1641 a broad panel of lectin probes 1629:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 10 35 from components 2302:2311 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 1 36 theme Classic 129:135 arg1 CG 151:152 CG 151:152 CG 151:152 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 36 theme Classic 129:135 arg1 galactosemia 137:148 Classic galactosemia 129:148 Classic galactosemia (CG) 129:153 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 36 theme Classic 129:135 arg1 disorder 181:188 an autosomal recessive disorder 158:188 an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis 158:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 10 37 theme altered 2261:2267 arg1 components 2302:2311 altered trans-synaptic signaling pathway components 2261:2311 altered trans-synaptic signaling pathway components 2261:2311 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 0 38 theme disease 114:120 arg1 model 122:126 a Drosophila classic galactosemia disease model 80:126 a Drosophila classic galactosemia disease model 80:126 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 8 39 theme profound 1668:1675 arg1 alterations 1677:1687 profound alterations 1668:1687 profound alterations 1668:1687 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 8 39 theme profound 1668:1675 arg1 depletion 1717:1725 depletion 1717:1725 depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1717:1816 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 2 40 theme glycoproteins 554:566 arg1 synthesis 541:549 the synthesis 537:549 the synthesis of glycoproteins and glycolipids 537:582 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 6 41 theme GALT 1201:1204 arg1 Loss 1182:1185 Loss 1182:1185 Loss of Drosophila GALT (dGALT) 1182:1212 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 0 42 theme Drosophila 82:91 arg1 model 122:126 a Drosophila classic galactosemia disease model 80:126 a Drosophila classic galactosemia disease model 80:126 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 0 43 theme Overelaborated 0:13 arg1 architecture 24:35 Overelaborated synaptic architecture 0:35 Overelaborated synaptic architecture 0:35 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 1 44 theme diphosphate 325:335 arg1 UDP 338:340 uridine diphosphate (UDP) 317:341 uridine diphosphate (UDP) 317:341 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 5 45 theme behavioral 1055:1064 arg1 traits 1066:1071 behavioral traits 1055:1071 behavioral traits 1055:1071 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 5 46 gly glycosylated 1099:1110 arg1 formation 1126:1134 glycosylated synaptomatrix formation 1099:1134 glycosylated synaptomatrix formation 1099:1134 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 3 47 theme striking 745:752 arg1 impairments 794:804 striking neurodevelopmental, motor and cognitive impairments 745:804 striking neurodevelopmental, motor and cognitive impairments 745:804 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 1 48 theme UDP 338:340 arg1 conversion 277:286 conversion 277:286 conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis 277:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 8 49 gly fucosylated 1768:1778 arg1 HRP 1804:1806 HRP 1804:1806 HRP 1804:1806 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 8 49 gly fucosylated 1768:1778 arg1 peroxidase 1792:1801 fucosylated horseradish peroxidase 1768:1801 fucosylated horseradish peroxidase (HRP) 1768:1807 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 7 50 theme dehydrogenase 1433:1445 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 50 theme dehydrogenase 1433:1445 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 8 51 with synaptomatrix 1610:1622 arg1 panel 1637:1641 a broad panel 1629:1641 a broad panel of lectin probes 1629:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 5 52 theme Drosophila 1156:1165 arg1 model 1175:1179 a GALT-deficient Drosophila disease model 1139:1179 a GALT-deficient Drosophila disease model 1139:1179 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 9 53 theme sulfate 2066:2072 arg1 HSPG 2088:2091 HSPG 2088:2091 HSPG 2088:2091 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 9 53 theme sulfate 2066:2072 arg1 proteoglycan 2074:2085 heparan sulfate proteoglycan 2058:2085 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 2 54 gly glycoproteins 554:566 arg1 glycoproteins 554:566 glycoproteins 554:566 glycoproteins 554:566 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 7 55 theme defects 1570:1576 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 55 theme defects 1570:1576 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 5 56 theme glycosylated 1099:1110 arg1 formation 1126:1134 glycosylated synaptomatrix formation 1099:1134 glycosylated synaptomatrix formation 1099:1134 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 6 57 theme coordinated 1230:1240 arg1 movement 1242:1249 coordinated movement 1230:1249 coordinated movement 1230:1249 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 7 58 theme galactokinase 1396:1408 arg1 dGALK 1411:1415 dGALK 1411:1415 dGALK 1411:1415 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 58 theme galactokinase 1396:1408 arg1 mutation 1384:1391 mutation 1384:1391 mutation of galactokinase 1384:1408 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 58 theme galactokinase 1396:1408 arg1 sugarless 1448:1456 sugarless 1448:1456 sugarless 1448:1456 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 58 theme galactokinase 1396:1408 arg1 dehydrogenase 1433:1445 UDP-glucose dehydrogenase 1421:1445 UDP-glucose dehydrogenase (sugarless) 1421:1457 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 58 theme galactokinase 1396:1408 arg1 galactose 1370:1378 Dietary galactose 1362:1378 Dietary galactose 1362:1378 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 10 59 theme coordinated 2352:2362 arg1 movement 2364:2371 impaired coordinated movement 2343:2371 impaired coordinated movement 2343:2371 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 8 60 theme horseradish 1780:1790 arg1 HRP 1804:1806 HRP 1804:1806 HRP 1804:1806 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 8 60 theme horseradish 1780:1790 arg1 peroxidase 1792:1801 fucosylated horseradish peroxidase 1768:1801 fucosylated horseradish peroxidase (HRP) 1768:1807 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 7 61 theme mutation 1384:1391 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 61 theme mutation 1384:1391 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 8 62 theme dGALK 1857:1861 arg1 co-removal 1863:1872 dGALK co-removal 1857:1872 dGALK co-removal 1857:1872 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 10 63 theme CG 2378:2379 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 6 64 theme architectural 1297:1309 arg1 abnormalities 1311:1323 architectural abnormalities 1297:1323 architectural abnormalities 1297:1323 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 3 65 with individuals 717:727 arg1 CG 734:735 CG 734:735 CG 734:735 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 4 66 theme O-linked 1015:1022 arg1 glycans 1024:1030 O-linked glycans 1015:1030 O-linked glycans 1015:1030 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 8 67 theme NMJ 1592:1594 arg1 synaptomatrix 1610:1622 the NMJ extracellular synaptomatrix 1588:1622 the NMJ extracellular synaptomatrix with a broad panel of lectin probes 1588:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 3 68 dep acute 663:667 arg1 lethal 682:687 lethal 682:687 lethal 682:687 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 10 69 theme disease 2394:2400 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 1 70 theme UDP-N-acetylglucosamine 447:469 arg1 synthesis 471:479 UDP-N-acetylglucosamine synthesis 447:479 UDP-N-acetylglucosamine synthesis 447:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 71 theme uridyltransferase 235:251 arg1 loss 205:208 loss 205:208 loss 205:208 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 10 72 theme defective 2314:2322 arg1 synaptogenesis 2324:2337 defective synaptogenesis 2314:2337 defective synaptogenesis 2314:2337 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 7 73 theme genetic 1525:1531 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 73 theme genetic 1525:1531 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 9 74 theme dGALK 2157:2161 arg1 co-removal 2163:2172 dGALK co-removal 2157:2172 dGALK co-removal 2157:2172 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 7 75 theme environmental 1507:1519 arg1 genes 1459:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes 1362:1463 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 75 theme environmental 1507:1519 arg1 modifiers 1533:1541 critical environmental and genetic modifiers 1498:1541 critical environmental and genetic modifiers of behavioral and cellular defects 1498:1576 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 3 76 theme maturing 708:715 arg1 individuals 717:727 maturing individuals 708:727 maturing individuals with CG 708:735 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 9 77 theme trans-synaptic 1929:1942 arg1 signals 1944:1950 trans-synaptic signals 1929:1950 trans-synaptic signals modulated by this synaptomatrix carbohydrate environment 1929:2007 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 10 78 theme glycosylation 2239:2251 arg1 losses 2253:2258 synaptomatrix glycosylation losses 2225:2258 synaptomatrix glycosylation losses 2225:2258 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 7 79 theme Dietary 1362:1368 arg1 dGALK 1411:1415 dGALK 1411:1415 dGALK 1411:1415 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 79 theme Dietary 1362:1368 arg1 galactose 1370:1378 Dietary galactose 1362:1378 Dietary galactose 1362:1378 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 8 80 theme lectin 1646:1651 arg1 probes 1653:1658 lectin probes 1646:1658 lectin probes 1646:1658 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 6 81 theme neuromuscular 1332:1344 arg1 NMJ 1356:1358 NMJ 1356:1358 NMJ 1356:1358 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 6 81 theme neuromuscular 1332:1344 arg1 junction 1346:1353 the neuromuscular junction 1328:1353 the neuromuscular junction (NMJ) 1328:1359 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 6 82 from junction 1346:1353 arg1 overelaboration 1277:1291 structural overelaboration 1266:1291 structural overelaboration 1266:1291 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 6 82 from junction 1346:1353 arg1 abnormalities 1311:1323 architectural abnormalities 1297:1323 architectural abnormalities 1297:1323 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 9 83 theme carbohydrate 1984:1995 arg1 environment 1997:2007 this synaptomatrix carbohydrate environment 1965:2007 this synaptomatrix carbohydrate environment 1965:2007 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 0 84 theme galactosemia 101:112 arg1 model 122:126 a Drosophila classic galactosemia disease model 80:126 a Drosophila classic galactosemia disease model 80:126 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 1 85 theme autosomal 161:169 arg1 disorder 181:188 an autosomal recessive disorder 158:188 an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis 158:479 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 1 85 theme autosomal 161:169 arg1 galactosemia 137:148 Classic galactosemia 129:148 Classic galactosemia (CG) 129:153 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 10 86 theme trans-synaptic 2269:2282 arg1 components 2302:2311 altered trans-synaptic signaling pathway components 2261:2311 altered trans-synaptic signaling pathway components 2261:2311 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 2 87 theme cell 608:611 arg1 surfaces 613:620 cell surfaces 608:620 cell surfaces 608:620 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 4 88 theme glycosylation 879:891 arg1 defects 893:899 glycosylation defects 879:899 glycosylation defects 879:899 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 5 89 theme synaptic 1074:1081 arg1 development 1083:1093 synaptic development 1074:1093 synaptic development 1074:1093 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 8 90 theme dGALT 1692:1696 arg1 mutants 1698:1704 dGALT mutants 1692:1704 dGALT mutants 1692:1704 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 0 91 theme synaptic 15:22 arg1 architecture 24:35 Overelaborated synaptic architecture 0:35 Overelaborated synaptic architecture 0:35 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 1 92 theme UDP-N-acetylgalactosamine 417:441 arg1 upstream 405:412 upstream 405:412 upstream 405:412 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 6 93 theme Drosophila 1190:1199 arg1 dGALT 1207:1211 dGALT 1207:1211 dGALT 1207:1211 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 6 93 theme Drosophila 1190:1199 arg1 GALT 1201:1204 Drosophila GALT 1190:1204 Drosophila GALT (dGALT) 1190:1212 Loss of Drosophila GALT (dGALT) greatly impairs coordinated movement and results in structural overelaboration and architectural abnormalities at the neuromuscular junction (NMJ). 25326312 9 94 theme dGALT-null 2014:2023 arg1 NMJs 2025:2028 dGALT-null NMJs 2014:2028 dGALT-null NMJs 2014:2028 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 10 95 from synaptogenesis 2324:2337 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 1 96 theme uridine 317:323 arg1 UDP 338:340 uridine diphosphate (UDP) 317:341 uridine diphosphate (UDP) 317:341 Classic galactosemia (CG) is an autosomal recessive disorder resulting from loss of galactose-1-phosphate uridyltransferase (GALT), which catalyzes conversion of galactose-1-phosphate and uridine diphosphate (UDP)-glucose to glucose-1-phosphate and UDP-galactose, immediately upstream of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine synthesis. 25326312 9 97 dep co-receptor 2094:2104 arg1 levels 2121:2126 levels 2121:2126 levels 2121:2126 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 2 98 theme essential 508:516 arg1 UDP-sugars 493:502 These four UDP-sugars 482:502 These four UDP-sugars 482:502 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 2 98 theme essential 508:516 arg1 donors 518:523 essential donors 508:523 essential donors 508:523 These four UDP-sugars are essential donors for driving the synthesis of glycoproteins and glycolipids, which heavily decorate cell surfaces and extracellular spaces. 25326312 0 99 theme synaptomatrix 49:61 arg1 glycosylation 63:75 synaptomatrix glycosylation 49:75 synaptomatrix glycosylation 49:75 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 10 100 from losses 2253:2258 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 7 101 theme UDP-glucose 1421:1431 arg1 sugarless 1448:1456 sugarless 1448:1456 sugarless 1448:1456 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 7 101 theme UDP-glucose 1421:1431 arg1 dehydrogenase 1433:1445 UDP-glucose dehydrogenase 1421:1445 UDP-glucose dehydrogenase (sugarless) 1421:1457 Dietary galactose and mutation of galactokinase (dGALK) or UDP-glucose dehydrogenase (sugarless) genes are identified, respectively, as critical environmental and genetic modifiers of behavioral and cellular defects. 25326312 0 102 gly glycosylation 63:75 arg1 model 122:126 a Drosophila classic galactosemia disease model 80:126 a Drosophila classic galactosemia disease model 80:126 Overelaborated synaptic architecture and reduced synaptomatrix glycosylation in a Drosophila classic galactosemia disease model. 25326312 8 103 theme galactosyl 1730:1739 arg1 moieties 1809:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 10 104 from movement 2364:2371 arg1 model 2402:2406 a CG neurological disease model 2376:2406 a CG neurological disease model 2376:2406 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 8 105 from alterations 1677:1687 arg1 mutants 1698:1704 dGALT mutants 1692:1704 dGALT mutants 1692:1704 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 5 106 theme GALT-deficient 1141:1154 arg1 model 1175:1179 a GALT-deficient Drosophila disease model 1139:1179 a GALT-deficient Drosophila disease model 1139:1179 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 9 107 theme heparan 2058:2064 arg1 HSPG 2088:2091 HSPG 2088:2091 HSPG 2088:2091 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 9 107 theme heparan 2058:2064 arg1 proteoglycan 2074:2085 heparan sulfate proteoglycan 2058:2085 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 8 108 theme N-acetylgalactosamine 1742:1762 arg1 moieties 1809:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 3 109 theme motor 774:778 arg1 impairments 794:804 striking neurodevelopmental, motor and cognitive impairments 745:804 striking neurodevelopmental, motor and cognitive impairments 745:804 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 4 110 theme congenital 940:949 arg1 CG 907:908 CG 907:908 CG 907:908 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 4 110 theme congenital 940:949 arg1 disorder 951:958 a congenital disorder 938:958 a congenital disorder of glycosylation (CDG) 938:981 Previous studies suggest that neurological symptoms are associated with glycosylation defects, with CG recently being described as a congenital disorder of glycosylation (CDG), showing defects in both N- and O-linked glycans. 25326312 9 111 theme proteoglycan 2074:2085 arg1 co-receptor 2094:2104 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 heparan sulfate proteoglycan (HSPG) co-receptor 2058:2104 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 8 112 theme fucosylated 1768:1778 arg1 HRP 1804:1806 HRP 1804:1806 HRP 1804:1806 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 8 112 theme fucosylated 1768:1778 arg1 peroxidase 1792:1801 fucosylated horseradish peroxidase 1768:1801 fucosylated horseradish peroxidase (HRP) 1768:1807 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25326312 3 113 theme neurodevelopmental 754:771 arg1 impairments 794:804 striking neurodevelopmental, motor and cognitive impairments 745:804 striking neurodevelopmental, motor and cognitive impairments 745:804 In addition to acute, potentially lethal neonatal symptoms, maturing individuals with CG develop striking neurodevelopmental, motor and cognitive impairments. 25326312 9 114 theme striking 2038:2045 arg1 changes 2047:2053 striking changes 2038:2053 striking changes 2038:2053 Synaptogenesis relies on trans-synaptic signals modulated by this synaptomatrix carbohydrate environment, and dGALT-null NMJs display striking changes in heparan sulfate proteoglycan (HSPG) co-receptor and Wnt ligand levels, which are also corrected by dGALK co-removal and sugarless overexpression. 25326312 5 115 theme synaptomatrix 1112:1124 arg1 formation 1126:1134 glycosylated synaptomatrix formation 1099:1134 glycosylated synaptomatrix formation 1099:1134 Here, we characterize behavioral traits, synaptic development and glycosylated synaptomatrix formation in a GALT-deficient Drosophila disease model. 25326312 10 116 theme pathway 2294:2300 arg1 components 2302:2311 altered trans-synaptic signaling pathway components 2261:2311 altered trans-synaptic signaling pathway components 2261:2311 These results reveal synaptomatrix glycosylation losses, altered trans-synaptic signaling pathway components, defective synaptogenesis and impaired coordinated movement in a CG neurological disease model. 25326312 8 117 theme peroxidase 1792:1801 arg1 moieties 1809:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties 1730:1816 Assaying the NMJ extracellular synaptomatrix with a broad panel of lectin probes reveals profound alterations in dGALT mutants, including depletion of galactosyl, N-acetylgalactosamine and fucosylated horseradish peroxidase (HRP) moieties, which are differentially corrected by dGALK co-removal and sugarless overexpression. 25488666 6 0 theme LC3-II-detected 1002:1016 arg1 autophagosomes 1018:1031 LC3-II-detected autophagosomes 1002:1031 LC3-II-detected autophagosomes 1002:1031 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 9 1 dep focusing 1412:1419 arg1 Isoelectric 1400:1410 Isoelectric 1400:1410 Isoelectric 1400:1410 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 6 2 theme Pompe 909:913 arg1 muscle 924:929 Pompe skeletal muscle 909:929 Pompe skeletal muscle 909:929 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 10 3 attach presence 1660:1667 arg2 precursor 1700:1708 a trimannose complex glycan precursor 1672:1708 a trimannose complex glycan precursor 1672:1708 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 10 3 attach presence 1660:1667 arg1 iPSC-CMs 1719:1726 Pompe iPSC-CMs 1713:1726 Pompe iPSC-CMs 1713:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 10 3 attach presence 1660:1667 arg2 structures 1635:1644 multiantennary structures 1620:1644 multiantennary structures 1620:1644 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 8 4 theme mobility 1291:1298 arg1 forms 1300:1304 the higher mobility forms 1280:1304 the higher mobility forms of the LAMPs 1280:1317 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 3 5 theme iPS 598:600 arg1 cells 603:607 induced pluripotent stem (iPS) cells 572:607 induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM) 572:660 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 6 6 theme comparable 972:981 arg1 rates 993:997 comparable clearance rates 972:997 comparable clearance rates of LC3-II-detected autophagosomes 972:1031 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 11 7 theme hypertrophic 1840:1851 arg1 cardiomyopathies 1853:1868 hypertrophic cardiomyopathies 1840:1868 hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation 1840:1922 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 3 8 from patients 516:523 arg1 fibroblasts 499:509 fibroblasts 499:509 fibroblasts from patients with infantile-onset Pompe disease 499:558 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 3 9 theme stem 592:595 arg1 cells 603:607 induced pluripotent stem (iPS) cells 572:607 induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM) 572:660 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 8 10 theme LAMPs 1313:1317 arg1 forms 1300:1304 the higher mobility forms 1280:1304 the higher mobility forms of the LAMPs 1280:1317 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 4 11 theme glycogen-filled 726:740 arg1 lysosomes 742:750 pathognomonic glycogen-filled lysosomes 712:750 pathognomonic glycogen-filled lysosomes 712:750 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 6 12 theme Impaired 867:874 arg1 autophagy 876:884 Impaired autophagy 867:884 Impaired autophagy 867:884 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 1 13 theme complete 203:210 arg1 loss 212:215 the complete loss 199:215 the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 199:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 10 14 theme MALDI-TOF-MS 1544:1555 arg1 analysis 1557:1564 MALDI-TOF-MS analysis 1544:1564 MALDI-TOF-MS analysis of N-linked glycans 1544:1584 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 8 15 theme Pompe 1336:1340 arg1 iPSC-CMs 1342:1349 Pompe iPSC-CMs 1336:1349 Pompe iPSC-CMs 1336:1349 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 10 16 from diversity 1607:1615 arg1 iPSC-CMs 1719:1726 Pompe iPSC-CMs 1713:1726 Pompe iPSC-CMs 1713:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 4 17 theme GAA 695:697 arg1 activity 699:706 undetectable GAA activity 682:706 undetectable GAA activity 682:706 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 7 18 theme control 1187:1193 arg1 iPSC-CMs 1195:1202 control iPSC-CMs 1187:1202 control iPSC-CMs 1187:1202 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 1 19 theme glycogen-hydrolyzing 230:249 arg1 α-glucosidase 263:275 glycogen-hydrolyzing enzyme acid α-glucosidase 230:275 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 19 theme glycogen-hydrolyzing 230:249 arg1 GAA 278:280 GAA 278:280 GAA 278:280 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 10 20 link N-linked 1569:1576 arg1 glycans 1578:1584 N-linked glycans 1569:1584 N-linked glycans 1569:1584 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 1 21 theme Pompe 140:144 arg1 disease 146:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 21 theme Pompe 140:144 arg1 disorder 180:187 an autosomal recessive disorder 157:187 an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 157:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 11 22 contain has 1774:1776 arg2 abnormality 1798:1808 a glycan processing abnormality 1778:1808 a glycan processing abnormality 1778:1808 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 11 22 contain has 1774:1776 arg1 cardiomyopathy 1759:1772 Pompe cardiomyopathy 1753:1772 Pompe cardiomyopathy 1753:1772 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 11 23 located observed 1870:1877 arg1 disorders 1897:1905 the congenital disorders 1882:1905 the congenital disorders of glycosylation 1882:1922 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 11 23 located observed 1870:1877 arg2 cardiomyopathies 1853:1868 hypertrophic cardiomyopathies 1840:1868 hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation 1840:1922 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 7 24 theme electrophoretic 1148:1162 arg1 mobility 1164:1171 higher electrophoretic mobility 1141:1171 higher electrophoretic mobility 1141:1171 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 9 25 theme focusing 1412:1419 arg1 studies 1421:1427 Isoelectric focusing studies 1400:1427 Isoelectric focusing studies 1400:1427 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 5 26 theme engineered 840:849 arg1 tissue 859:864 engineered cardiac tissue 840:864 engineered cardiac tissue 840:864 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 1 27 theme cardiac 356:362 arg1 pathology 384:392 prominent cardiac and skeletal muscle pathology 346:392 prominent cardiac and skeletal muscle pathology 346:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 28 theme autosomal 160:168 arg1 disease 146:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 28 theme autosomal 160:168 arg1 disorder 180:187 an autosomal recessive disorder 157:187 an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 157:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 0 29 theme induced 70:76 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 1 30 theme skeletal 368:375 arg1 pathology 384:392 prominent cardiac and skeletal muscle pathology 346:392 prominent cardiac and skeletal muscle pathology 346:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 10 31 theme reduced 1599:1605 arg1 diversity 1607:1615 reduced diversity 1599:1615 reduced diversity of multiantennary structures 1599:1644 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 0 32 theme stem 90:93 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 7 33 theme lysosome-associated 1052:1070 arg1 LAMP2 1101:1105 LAMP2 1101:1105 LAMP2 1101:1105 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 33 theme lysosome-associated 1052:1070 arg1 LAMP1 1091:1095 LAMP1 1091:1095 LAMP1 1091:1095 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 33 theme lysosome-associated 1052:1070 arg1 proteins 1081:1088 the lysosome-associated membrane proteins 1048:1088 the lysosome-associated membrane proteins 1048:1088 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 9 34 theme alkaline 1460:1467 arg1 pI 1469:1470 a more alkaline pI 1453:1470 a more alkaline pI in Pompe 1453:1479 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 10 35 theme structures 1635:1644 arg1 diversity 1607:1615 reduced diversity 1599:1615 reduced diversity of multiantennary structures 1599:1644 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 10 35 theme structures 1635:1644 arg1 presence 1660:1667 the major presence 1650:1667 the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs 1650:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 0 36 theme Pompe 0:4 arg1 disease 6:12 Pompe disease 0:12 Pompe disease 0:12 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 8 37 theme Brefeldin 1205:1213 arg1 A 1215:1215 Brefeldin A 1205:1215 Brefeldin A 1205:1215 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 2 38 theme activity 430:437 arg1 loss 418:421 loss 418:421 loss of GAA activity 418:437 The mechanism by which loss of GAA activity causes cardiomyopathy is poorly understood. 25488666 1 39 theme glycogen 320:327 arg1 accumulation 329:340 lysosomal glycogen accumulation 310:340 lysosomal glycogen accumulation 310:340 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 9 40 theme control 1495:1501 arg1 iPSC-CMs 1503:1510 control iPSC-CMs 1495:1510 control iPSC-CMs due largely to hyposialylation 1495:1541 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 10 41 theme precursor 1700:1708 arg1 diversity 1607:1615 reduced diversity 1599:1615 reduced diversity of multiantennary structures 1599:1644 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 10 41 theme precursor 1700:1708 arg1 presence 1660:1667 the major presence 1650:1667 the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs 1650:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 4 42 contain had 678:680 arg2 lysosomes 742:750 pathognomonic glycogen-filled lysosomes 712:750 pathognomonic glycogen-filled lysosomes 712:750 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 4 42 contain had 678:680 arg1 iPSC-CMs 669:676 Pompe iPSC-CMs 663:676 Pompe iPSC-CMs 663:676 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 4 42 contain had 678:680 arg2 activity 699:706 undetectable GAA activity 682:706 undetectable GAA activity 682:706 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 3 43 theme infantile-onset 530:544 arg1 disease 552:558 infantile-onset Pompe disease 530:558 infantile-onset Pompe disease 530:558 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 0 44 theme glycosylation 39:51 arg1 deficit 53:59 a Golgi-based glycosylation deficit 25:59 a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes 25:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 9 45 theme due 1512:1514 arg1 iPSC-CMs 1503:1510 control iPSC-CMs 1495:1510 control iPSC-CMs due largely to hyposialylation 1495:1541 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 10 46 theme complex 1685:1691 arg1 precursor 1700:1708 a trimannose complex glycan precursor 1672:1708 a trimannose complex glycan precursor 1672:1708 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 10 47 theme glycans 1578:1584 arg1 analysis 1557:1564 MALDI-TOF-MS analysis 1544:1564 MALDI-TOF-MS analysis of N-linked glycans 1544:1584 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 1 48 theme enzyme 251:256 arg1 α-glucosidase 263:275 glycogen-hydrolyzing enzyme acid α-glucosidase 230:275 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 48 theme enzyme 251:256 arg1 GAA 278:280 GAA 278:280 GAA 278:280 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 8 49 theme Golgi 1243:1247 arg1 disruption 1225:1234 disruption 1225:1234 disruption of the Golgi 1225:1247 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 1 50 theme α-glucosidase 263:275 arg1 activity 283:290 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 8 51 theme control 1252:1258 arg1 iPSC-CMs 1260:1267 control iPSC-CMs 1252:1267 control iPSC-CMs reproduced the higher mobility forms of the LAMPs 1252:1317 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 6 52 theme autophagosomes 1018:1031 arg1 rates 993:997 comparable clearance rates 972:997 comparable clearance rates of LC3-II-detected autophagosomes 972:1031 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 0 53 link cell-derived 95:106 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 10 54 theme Pompe 1713:1717 arg1 iPSC-CMs 1719:1726 Pompe iPSC-CMs 1713:1726 Pompe iPSC-CMs 1713:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 5 55 theme cardiac 851:857 arg1 tissue 859:864 engineered cardiac tissue 840:864 engineered cardiac tissue 840:864 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 11 56 theme processing 1787:1796 arg1 abnormality 1798:1808 a glycan processing abnormality 1778:1808 a glycan processing abnormality 1778:1808 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 11 57 theme congenital 1886:1895 arg1 disorders 1897:1905 the congenital disorders 1882:1905 the congenital disorders of glycosylation 1882:1922 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 1 58 theme activity 283:290 arg1 loss 212:215 the complete loss 199:215 the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 199:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 8 59 theme higher 1284:1289 arg1 forms 1300:1304 the higher mobility forms 1280:1304 the higher mobility forms of the LAMPs 1280:1317 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 6 60 theme clearance 983:991 arg1 rates 993:997 comparable clearance rates 972:997 comparable clearance rates of LC3-II-detected autophagosomes 972:1031 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 3 61 with patients 516:523 arg1 disease 552:558 infantile-onset Pompe disease 530:558 infantile-onset Pompe disease 530:558 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 3 62 theme pluripotent 580:590 arg1 cells 603:607 induced pluripotent stem (iPS) cells 572:607 induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM) 572:660 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 10 63 theme N-linked 1569:1576 arg1 glycans 1578:1584 N-linked glycans 1569:1584 N-linked glycans 1569:1584 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 9 64 from pI 1469:1470 arg1 Pompe 1475:1479 Pompe 1475:1479 Pompe 1475:1479 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 8 65 theme appropriate 1373:1383 arg1 glycosylation 1385:1397 appropriate glycosylation 1373:1397 appropriate glycosylation 1373:1397 Brefeldin A induced disruption of the Golgi in control iPSC-CMs reproduced the higher mobility forms of the LAMPs, suggesting that Pompe iPSC-CMs produce LAMPs lacking appropriate glycosylation. 25488666 5 66 theme Pompe 766:770 arg1 iPSC-CMs 784:791 Pompe and control iPSC-CMs 766:791 iPSC-CMs 784:791 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 6 67 contain had 968:970 arg1 control 941:947 control 941:947 control 941:947 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 6 67 contain had 968:970 arg2 rates 993:997 comparable clearance rates 972:997 comparable clearance rates of LC3-II-detected autophagosomes 972:1031 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 6 67 contain had 968:970 arg1 Pompe 953:957 Pompe 953:957 Pompe 953:957 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 5 68 theme control 776:782 arg1 iPSC-CMs 784:791 Pompe and control iPSC-CMs 766:791 iPSC-CMs 784:791 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 4 69 theme pathognomonic 712:724 arg1 lysosomes 742:750 pathognomonic glycogen-filled lysosomes 712:750 pathognomonic glycogen-filled lysosomes 712:750 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 1 70 theme lysosomal 220:228 arg1 activity 283:290 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 4 71 theme undetectable 682:693 arg1 activity 699:706 undetectable GAA activity 682:706 undetectable GAA activity 682:706 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 1 72 theme Infantile-onset 124:138 arg1 disease 146:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 72 theme Infantile-onset 124:138 arg1 disorder 180:187 an autosomal recessive disorder 157:187 an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 157:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 7 73 theme higher 1141:1146 arg1 mobility 1164:1171 higher electrophoretic mobility 1141:1171 higher electrophoretic mobility 1141:1171 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 74 theme Pompe 1113:1117 arg1 iPSC-CMs 1119:1126 Pompe iPSC-CMs 1113:1126 Pompe iPSC-CMs 1113:1126 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 10 75 from presence 1660:1667 arg1 iPSC-CMs 1719:1726 Pompe iPSC-CMs 1713:1726 Pompe iPSC-CMs 1713:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 0 76 theme human 64:68 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 5 77 theme comparable 803:812 arg1 properties 826:835 comparable contractile properties 803:835 comparable contractile properties 803:835 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 0 78 theme pluripotent 78:88 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 1 79 theme recessive 170:178 arg1 disease 146:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease 124:152 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 79 theme recessive 170:178 arg1 disorder 180:187 an autosomal recessive disorder 157:187 an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology 157:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 0 80 theme cell-derived 95:106 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 1 81 theme muscle 377:382 arg1 pathology 384:392 prominent cardiac and skeletal muscle pathology 346:392 prominent cardiac and skeletal muscle pathology 346:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 0 82 from deficit 53:59 arg1 cardiomyocytes 108:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 human induced pluripotent stem cell-derived cardiomyocytes 64:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 9 83 contain has 1449:1451 arg1 LAMP2 1443:1447 LAMP2 1443:1447 LAMP2 1443:1447 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 9 83 contain has 1449:1451 arg2 pI 1469:1470 a more alkaline pI 1453:1470 a more alkaline pI in Pompe 1453:1479 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 5 84 theme contractile 814:824 arg1 properties 826:835 comparable contractile properties 803:835 comparable contractile properties 803:835 Nonetheless, Pompe and control iPSC-CMs exhibited comparable contractile properties in engineered cardiac tissue. 25488666 9 85 theme more 1455:1458 arg1 pI 1469:1470 a more alkaline pI 1453:1470 a more alkaline pI in Pompe 1453:1479 Isoelectric focusing studies revealed that LAMP2 has a more alkaline pI in Pompe compared with control iPSC-CMs due largely to hyposialylation. 25488666 7 86 theme membrane 1072:1079 arg1 LAMP2 1101:1105 LAMP2 1101:1105 LAMP2 1101:1105 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 86 theme membrane 1072:1079 arg1 LAMP1 1091:1095 LAMP1 1091:1095 LAMP1 1091:1095 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 86 theme membrane 1072:1079 arg1 proteins 1081:1088 the lysosome-associated membrane proteins 1048:1088 the lysosome-associated membrane proteins 1048:1088 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 10 87 theme multiantennary 1620:1633 arg1 structures 1635:1644 multiantennary structures 1620:1644 multiantennary structures 1620:1644 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 0 88 theme Golgi-based 27:37 arg1 deficit 53:59 a Golgi-based glycosylation deficit 25:59 a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes 25:121 Pompe disease results in a Golgi-based glycosylation deficit in human induced pluripotent stem cell-derived cardiomyocytes. 25488666 1 89 theme lysosomal 310:318 arg1 accumulation 329:340 lysosomal glycogen accumulation 310:340 lysosomal glycogen accumulation 310:340 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 7 90 from iPSC-CMs 1119:1126 arg1 LAMP2 1101:1105 LAMP2 1101:1105 LAMP2 1101:1105 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 90 from iPSC-CMs 1119:1126 arg1 LAMP1 1091:1095 LAMP1 1091:1095 LAMP1 1091:1095 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 7 90 from iPSC-CMs 1119:1126 arg1 proteins 1081:1088 the lysosome-associated membrane proteins 1048:1088 the lysosome-associated membrane proteins 1048:1088 Unexpectedly, the lysosome-associated membrane proteins, LAMP1 and LAMP2, from Pompe iPSC-CMs demonstrated higher electrophoretic mobility compared with control iPSC-CMs. 25488666 6 91 dep control 941:947 arg1 iPSC-CMs 959:966 iPSC-CMs 959:966 iPSC-CMs 959:966 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 10 92 theme major 1654:1658 arg1 presence 1660:1667 the major presence 1650:1667 the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs 1650:1726 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 2 93 theme GAA 426:428 arg1 activity 430:437 GAA activity 426:437 GAA activity 426:437 The mechanism by which loss of GAA activity causes cardiomyopathy is poorly understood. 25488666 10 94 theme glycan 1693:1698 arg1 precursor 1700:1708 a trimannose complex glycan precursor 1672:1708 a trimannose complex glycan precursor 1672:1708 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 11 95 theme Pompe 1753:1757 arg1 cardiomyopathy 1759:1772 Pompe cardiomyopathy 1753:1772 Pompe cardiomyopathy 1753:1772 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 3 96 theme Pompe 546:550 arg1 disease 552:558 infantile-onset Pompe disease 530:558 infantile-onset Pompe disease 530:558 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 25488666 1 97 theme prominent 346:354 arg1 pathology 384:392 prominent cardiac and skeletal muscle pathology 346:392 prominent cardiac and skeletal muscle pathology 346:392 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 10 98 theme trimannose 1674:1683 arg1 precursor 1700:1708 a trimannose complex glycan precursor 1672:1708 a trimannose complex glycan precursor 1672:1708 MALDI-TOF-MS analysis of N-linked glycans demonstrated reduced diversity of multiantennary structures and the major presence of a trimannose complex glycan precursor in Pompe iPSC-CMs. 25488666 11 99 theme glycosylation 1910:1922 arg1 disorders 1897:1905 the congenital disorders 1882:1905 the congenital disorders of glycosylation 1882:1922 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 1 100 theme acid 258:261 arg1 α-glucosidase 263:275 glycogen-hydrolyzing enzyme acid α-glucosidase 230:275 lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity 220:290 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 1 100 theme acid 258:261 arg1 GAA 278:280 GAA 278:280 GAA 278:280 Infantile-onset Pompe disease is an autosomal recessive disorder caused by the complete loss of lysosomal glycogen-hydrolyzing enzyme acid α-glucosidase (GAA) activity, which results in lysosomal glycogen accumulation and prominent cardiac and skeletal muscle pathology. 25488666 6 101 theme skeletal 915:922 arg1 muscle 924:929 Pompe skeletal muscle 909:929 Pompe skeletal muscle 909:929 Impaired autophagy has been implicated in Pompe skeletal muscle; however, control and Pompe iPSC-CMs had comparable clearance rates of LC3-II-detected autophagosomes. 25488666 4 102 theme Pompe 663:667 arg1 iPSC-CMs 669:676 Pompe iPSC-CMs 663:676 Pompe iPSC-CMs 663:676 Pompe iPSC-CMs had undetectable GAA activity and pathognomonic glycogen-filled lysosomes. 25488666 11 103 theme glycan 1780:1785 arg1 abnormality 1798:1808 a glycan processing abnormality 1778:1808 a glycan processing abnormality 1778:1808 These data suggest that Pompe cardiomyopathy has a glycan processing abnormality and thus shares features with hypertrophic cardiomyopathies observed in the congenital disorders of glycosylation. 25488666 3 104 theme induced 572:578 arg1 cells 603:607 induced pluripotent stem (iPS) cells 572:607 induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM) 572:660 We reprogrammed fibroblasts from patients with infantile-onset Pompe disease to generate induced pluripotent stem (iPS) cells that were differentiated to cardiomyocytes (iPSC-CM). 26208585 0 0 theme plant 92:96 arg1 walls 103:107 plant cell walls 92:107 plant cell walls 92:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 5 1 from present 1086:1092 arg1 heteroxylans 1097:1108 heteroxylans 1097:1108 heteroxylans 1097:1108 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 4 2 theme rhamnogalacturonan-I 897:916 arg1 oligosaccharides 925:940 rhamnogalacturonan-I (RG-I) oligosaccharides 897:940 rhamnogalacturonan-I (RG-I) oligosaccharides 897:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 7 3 theme spelt 1285:1289 arg1 xylan 1291:1295 oat spelt xylan 1281:1295 oat spelt xylan 1281:1295 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 5 4 from walls 1021:1025 arg1 preparations 992:1003 heteroxylan preparations 980:1003 heteroxylan preparations from grass cell walls 980:1025 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 7 5 theme oat 1281:1283 arg1 xylan 1291:1295 oat spelt xylan 1281:1295 oat spelt xylan 1281:1295 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 3 6 theme oligosaccharide 724:738 arg1 structures 740:749 oligosaccharide structures 724:749 oligosaccharide structures 724:749 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 6 7 theme potato 1152:1157 arg1 walls 1170:1174 potato tuber cell walls 1152:1174 potato tuber cell walls 1152:1174 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 0 8 theme walls 103:107 arg1 glycans 81:87 heteroxylan and other glycans 59:87 heteroxylan and other glycans of plant cell walls 59:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 3 9 theme structures 740:749 arg1 recognition 709:719 the specific recognition 696:719 the specific recognition of oligosaccharide structures 696:749 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 4 10 theme RG-I 919:922 arg1 oligosaccharides 925:940 rhamnogalacturonan-I (RG-I) oligosaccharides 897:940 rhamnogalacturonan-I (RG-I) oligosaccharides 897:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 4 11 theme potato 859:864 arg1 fraction 876:883 a potato cell wall fraction 857:883 a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides 857:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 3 12 theme Monoclonal 583:592 arg1 MAbs 606:609 MAbs 606:609 MAbs 606:609 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 3 12 theme Monoclonal 583:592 arg1 antibodies 594:603 Monoclonal antibodies 583:603 Monoclonal antibodies (MAbs) 583:610 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 3 12 theme Monoclonal 583:592 arg1 tools 626:630 versatile tools 616:630 versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures 616:749 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 8 13 theme molecular 1539:1547 arg1 tools 1549:1553 molecular tools 1539:1553 molecular tools for the further analysis of such links 1539:1592 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 0 14 theme cell 98:101 arg1 walls 103:107 plant cell walls 92:107 plant cell walls 92:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 1 15 theme oat 392:394 arg1 walls 407:411 oat grain cell walls 392:411 oat grain cell walls 392:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 5 16 theme grass 1010:1014 arg1 walls 1021:1025 grass cell walls 1010:1025 grass cell walls 1010:1025 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 6 17 theme cell 1165:1168 arg1 walls 1170:1174 potato tuber cell walls 1152:1174 potato tuber cell walls 1152:1174 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 2 18 theme inter-linkages 539:552 arg1 terms 517:521 terms 517:521 terms of both in muro inter-linkages and developmental functions 517:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 1 19 theme grain 396:400 arg1 walls 407:411 oat grain cell walls 392:411 oat grain cell walls 392:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 4 20 theme wall 871:874 arg1 fraction 876:883 a potato cell wall fraction 857:883 a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides 857:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 5 21 theme cell 1016:1019 arg1 walls 1021:1025 grass cell walls 1010:1025 grass cell walls 1010:1025 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 2 22 theme Plant 414:418 arg1 glycans 479:485 structurally distinct glycans 457:485 structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 457:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 22 theme Plant 414:418 arg1 walls 425:429 Plant cell walls 414:429 Plant cell walls 414:429 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 22 theme Plant 414:418 arg1 composites 443:452 complex composites 435:452 complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 435:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 1 23 theme heteroxylan 176:186 arg1 preparations 213:224 heteroxylan cell wall polysaccharide preparations 176:224 heteroxylan cell wall polysaccharide preparations 176:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 1 24 theme cell 402:405 arg1 walls 407:411 oat grain cell walls 392:411 oat grain cell walls 392:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 4 25 theme cell 866:869 arg1 fraction 876:883 a potato cell wall fraction 857:883 a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides 857:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 8 26 theme links 1588:1592 arg1 analysis 1571:1578 the further analysis 1559:1578 the further analysis of such links 1559:1592 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 2 27 theme glycans 479:485 arg1 glycans 479:485 structurally distinct glycans 457:485 structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 457:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 27 theme glycans 479:485 arg1 walls 425:429 Plant cell walls 414:429 Plant cell walls 414:429 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 27 theme glycans 479:485 arg1 composites 443:452 complex composites 435:452 complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 435:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 1 28 theme cell 188:191 arg1 preparations 213:224 heteroxylan cell wall polysaccharide preparations 176:224 heteroxylan cell wall polysaccharide preparations 176:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 5 29 from heteroxylans 1097:1108 arg1 present 1086:1092 present 1086:1092 present 1086:1092 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 8 30 theme such 1583:1586 arg1 links 1588:1592 such links 1583:1592 such links 1583:1592 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 1 31 theme wall 193:196 arg1 preparations 213:224 heteroxylan cell wall polysaccharide preparations 176:224 heteroxylan cell wall polysaccharide preparations 176:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 0 32 theme Monoclonal 0:9 arg1 antibodies 11:20 Monoclonal antibodies 0:20 Monoclonal antibodies 0:20 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 4 33 theme MAbs 779:782 arg1 isolation 756:764 The isolation 752:764 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, 752:941 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 1 34 theme polysaccharide 198:211 arg1 preparations 213:224 heteroxylan cell wall polysaccharide preparations 176:224 heteroxylan cell wall polysaccharide preparations 176:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 0 35 theme low-abundance 31:43 arg1 links 45:49 low-abundance links 31:49 low-abundance links between heteroxylan and other glycans of plant cell walls 31:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 1 36 theme potato 313:318 arg1 walls 331:335 potato tuber cell walls 313:335 potato tuber cell walls 313:335 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 7 37 theme arabinogalactan-protein 1390:1412 arg1 epitope 1414:1420 the LM2 arabinogalactan-protein epitope 1382:1420 the LM2 arabinogalactan-protein epitope 1382:1420 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 5 38 theme heteroxylan 980:990 arg1 preparations 992:1003 heteroxylan preparations 980:1003 heteroxylan preparations from grass cell walls 980:1025 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 7 39 dep xylan 1291:1295 arg1 both 1297:1300 both 1297:1300 both 1297:1300 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 1 40 theme tuber 320:324 arg1 walls 331:335 potato tuber cell walls 313:335 potato tuber cell walls 313:335 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 1 41 from pectin 303:308 arg1 walls 331:335 potato tuber cell walls 313:335 potato tuber cell walls 313:335 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 8 42 theme cell 1514:1517 arg1 walls 1519:1523 plant cell walls 1508:1523 plant cell walls 1508:1523 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 5 43 attach present 1086:1092 arg1 heteroxylans 1097:1108 heteroxylans 1097:1108 heteroxylans 1097:1108 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 5 43 attach present 1086:1092 arg2 epitope 1071:1077 a glucuronosyl-containing epitope 1045:1077 a glucuronosyl-containing epitope widely present in heteroxylans 1045:1108 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 1 44 from arabinogalactan-proteins 364:387 arg1 walls 407:411 oat grain cell walls 392:411 oat grain cell walls 392:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 1 45 theme cell 326:329 arg1 walls 331:335 potato tuber cell walls 313:335 potato tuber cell walls 313:335 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 8 46 theme plant 1508:1512 arg1 walls 1519:1523 plant cell walls 1508:1523 plant cell walls 1508:1523 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 2 47 theme complex 435:441 arg1 glycans 479:485 structurally distinct glycans 457:485 structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 457:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 47 theme complex 435:441 arg1 walls 425:429 Plant cell walls 414:429 Plant cell walls 414:429 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 47 theme complex 435:441 arg1 composites 443:452 complex composites 435:452 complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 435:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 8 48 theme potential 1458:1466 arg1 occurrence 1468:1477 the potential occurrence 1454:1477 the potential occurrence of inter-glycan links within plant cell walls 1454:1523 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 2 49 theme functions 572:580 arg1 terms 517:521 terms 517:521 terms of both in muro inter-linkages and developmental functions 517:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 7 50 attach linked 1333:1338 arg2 epitopes 1320:1327 LM28 epitopes 1315:1327 LM28 epitopes 1315:1327 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 7 50 attach linked 1333:1338 arg1 arabinogalactan-proteins 1343:1366 arabinogalactan-proteins 1343:1366 arabinogalactan-proteins 1343:1366 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 2 51 theme distinct 470:477 arg1 glycans 479:485 structurally distinct glycans 457:485 structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 457:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 1 52 from xylan 354:358 arg1 walls 407:411 oat grain cell walls 392:411 oat grain cell walls 392:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 2 53 theme developmental 558:570 arg1 functions 572:580 developmental functions 558:580 developmental functions 558:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 3 54 theme high 671:674 arg1 sensitivity 676:686 high sensitivity 671:686 high sensitivity 671:686 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 3 55 theme versatile 616:624 arg1 antibodies 594:603 Monoclonal antibodies 583:603 Monoclonal antibodies (MAbs) 583:610 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 3 55 theme versatile 616:624 arg1 tools 626:630 versatile tools 616:630 versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures 616:749 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 6 56 theme tuber 1159:1163 arg1 walls 1170:1174 potato tuber cell walls 1152:1174 potato tuber cell walls 1152:1174 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 4 57 with immunisation 839:850 arg1 fraction 876:883 a potato cell wall fraction 857:883 a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides 857:940 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 0 58 theme heteroxylan 59:69 arg1 glycans 81:87 heteroxylan and other glycans 59:87 heteroxylan and other glycans of plant cell walls 59:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 8 59 theme occurrence 1468:1477 arg1 knowledge 1441:1449 knowledge 1441:1449 knowledge of the potential occurrence of inter-glycan links within plant cell walls 1441:1523 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 5 60 theme present 1086:1092 arg1 epitope 1071:1077 a glucuronosyl-containing epitope 1045:1077 a glucuronosyl-containing epitope widely present in heteroxylans 1045:1108 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 8 61 theme links 1495:1499 arg1 occurrence 1468:1477 the potential occurrence 1454:1477 the potential occurrence of inter-glycan links within plant cell walls 1454:1523 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 7 62 theme LM28 1315:1318 arg1 epitopes 1320:1327 LM28 epitopes 1315:1327 LM28 epitopes 1315:1327 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 8 63 theme inter-glycan 1482:1493 arg1 links 1495:1499 inter-glycan links 1482:1499 inter-glycan links 1482:1499 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 4 64 theme subsequent 825:834 arg1 heteroxylan 812:822 heteroxylan 812:822 heteroxylan 812:822 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 2 65 dep in 531:532 arg1 muro 534:537 muro 534:537 muro 534:537 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 6 66 theme pectic 1214:1219 arg1 macromolecules 1221:1234 pectic macromolecules 1214:1234 pectic macromolecules 1214:1234 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 3 67 theme wall 653:656 arg1 glycans 658:664 cell wall glycans 648:664 cell wall glycans 648:664 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 5 68 theme glucuronosyl-containing 1047:1069 arg1 epitope 1071:1077 a glucuronosyl-containing epitope 1045:1077 a glucuronosyl-containing epitope widely present in heteroxylans 1045:1108 LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. 26208585 4 69 theme novel 773:777 arg1 LM27 785:788 LM27 785:788 LM27 785:788 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 4 69 theme novel 773:777 arg1 MAbs 779:782 two novel MAbs 769:782 two novel MAbs 769:782 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 4 69 theme novel 773:777 arg1 LM28 794:797 LM28 794:797 LM28 794:797 The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. 26208585 1 70 theme sensitive 132:140 arg1 antibodies 153:162 two sensitive monoclonal antibodies 128:162 two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations 128:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 6 71 attach linked 1204:1209 arg2 glucuronoxylan 1182:1195 some glucuronoxylan 1177:1195 some glucuronoxylan 1177:1195 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 6 71 attach linked 1204:1209 arg1 macromolecules 1221:1234 pectic macromolecules 1214:1234 pectic macromolecules 1214:1234 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 6 71 attach linked 1204:1209 arg3 walls 1170:1174 potato tuber cell walls 1152:1174 potato tuber cell walls 1152:1174 Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. 26208585 2 72 theme in 531:532 arg1 inter-linkages 539:552 in muro inter-linkages 531:552 in muro inter-linkages 531:552 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 3 73 theme specific 700:707 arg1 recognition 709:719 the specific recognition 696:719 the specific recognition of oligosaccharide structures 696:749 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 7 74 theme LM2 1386:1388 arg1 epitope 1414:1420 the LM2 arabinogalactan-protein epitope 1382:1420 the LM2 arabinogalactan-protein epitope 1382:1420 Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. 26208585 1 75 theme monoclonal 142:151 arg1 antibodies 153:162 two sensitive monoclonal antibodies 128:162 two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations 128:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 2 76 theme cell 420:423 arg1 glycans 479:485 structurally distinct glycans 457:485 structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 457:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 76 theme cell 420:423 arg1 walls 425:429 Plant cell walls 414:429 Plant cell walls 414:429 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 2 76 theme cell 420:423 arg1 composites 443:452 complex composites 435:452 complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions 435:580 Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. 26208585 1 77 theme potential 260:268 arg1 inter-linkages 270:283 potential inter-linkages 260:283 potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls 260:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 0 78 theme other 75:79 arg1 glycans 81:87 heteroxylan and other glycans 59:87 heteroxylan and other glycans of plant cell walls 59:107 Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls. 26208585 8 79 theme further 1563:1569 arg1 analysis 1571:1578 the further analysis 1559:1578 the further analysis of such links 1559:1592 This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links. 26208585 3 80 theme cell 648:651 arg1 glycans 658:664 cell wall glycans 648:664 cell wall glycans 648:664 Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. 26208585 1 81 theme antibodies 153:162 arg1 derivation 114:123 The derivation 110:123 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations 110:224 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 26208585 1 82 theme inter-linkages 270:283 arg1 identification 242:255 the identification 238:255 the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls 238:411 The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. 24327722 10 0 located observed 1480:1487 arg1 ulcer 1501:1505 duodenal ulcer 1492:1505 duodenal ulcer 1492:1505 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 10 0 located observed 1480:1487 arg2 levels 1446:1451 Altered levels 1438:1451 Altered levels of serum glycans 1438:1468 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 9 1 theme glycans 1251:1257 arg1 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 2 theme glycans 1260:1266 arg1 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 1 3 theme biologic 100:107 arg1 systems 109:115 biologic systems 100:115 biologic systems 100:115 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 0 4 theme Serum 0:4 arg1 signatures 13:22 Serum glycan signatures 0:22 Serum glycan signatures of gastric cancer 0:40 Serum glycan signatures of gastric cancer. 24327722 12 5 theme cancer 1835:1840 arg1 pathology 1842:1850 gastric cancer pathology 1827:1850 gastric cancer pathology 1827:1850 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 5 6 with method 806:811 arg1 F 825:825 PNGase F 818:825 PNGase F 818:825 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 9 7 theme type 1285:1288 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 1 8 theme diagnostic 163:172 arg1 tool 174:177 a simple yet highly sensitive diagnostic tool 133:177 a simple yet highly sensitive diagnostic tool for disease onset and progression 133:211 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 12 9 theme Further 1705:1711 arg1 studies 1713:1719 Further studies 1705:1719 Further studies 1705:1719 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 3 10 with Patients 371:378 arg1 ulcer 394:398 duodenal ulcer 385:398 duodenal ulcer 385:398 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 4 11 theme serum 624:628 arg1 samples 630:636 72 serum samples 621:636 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer 621:742 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 9 12 theme biantennary 1316:1326 arg1 glycans 1328:1334 bigalactosylated biantennary glycans 1299:1334 bigalactosylated biantennary glycans 1299:1334 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 3 13 theme duodenal 592:599 arg1 ulcer 601:605 duodenal ulcer 592:605 duodenal ulcer 592:605 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 6 14 theme glycan 971:976 arg1 compositions 978:989 The corresponding glycan compositions 953:989 The corresponding glycan compositions 953:989 The corresponding glycan compositions were calculated based on accurate mass. 24327722 7 15 theme statistical 1043:1053 arg1 analysis 1055:1062 ANOVA-based statistical analysis 1031:1062 ANOVA-based statistical analysis 1031:1062 ANOVA-based statistical analysis was performed to identify potential markers for each subgroup. 24327722 5 16 theme mass 934:937 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 12 17 theme glycosylation 1810:1822 arg1 role 1794:1797 the role 1790:1797 the role of protein glycosylation in gastric cancer pathology 1790:1850 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 11 18 theme Serum 1580:1584 arg1 profiles 1593:1600 Serum glycan profiles 1580:1600 Serum glycan profiles 1580:1600 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 9 19 theme increased 1341:1349 arg1 levels 1351:1356 increased levels 1341:1356 increased levels of nongalactosylated biantennary glycans 1341:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 2 20 theme glycan 263:268 arg1 markers 270:276 glycan markers 263:276 glycan markers that would differentiate patients with gastric cancer from those with nonatrophic gastritis 263:368 This study aimed to use glycomics to investigate glycan markers that would differentiate patients with gastric cancer from those with nonatrophic gastritis. 24327722 11 21 from controls 1668:1675 arg1 cases 1657:1661 gastric cancer cases 1642:1661 gastric cancer cases from controls with nonatrophic gastritis 1642:1702 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 12 22 theme gastric 1827:1833 arg1 pathology 1842:1850 gastric cancer pathology 1827:1850 gastric cancer pathology 1827:1850 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 5 23 theme Fourier 892:898 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 4 24 theme nonatrophic 687:697 arg1 gastritis 699:707 nonatrophic gastritis 687:707 nonatrophic gastritis 687:707 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 9 25 theme biantennary 1379:1389 arg1 glycans 1391:1397 nongalactosylated biantennary glycans 1361:1397 nongalactosylated biantennary glycans 1361:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 1 26 theme simple 135:140 arg1 tool 174:177 a simple yet highly sensitive diagnostic tool 133:177 a simple yet highly sensitive diagnostic tool for disease onset and progression 133:211 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 6 27 theme accurate 1016:1023 arg1 mass 1025:1028 accurate mass 1016:1028 accurate mass 1016:1028 The corresponding glycan compositions were calculated based on accurate mass. 24327722 9 28 with levels 1351:1356 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 10 29 theme same 1546:1549 arg1 direction 1551:1559 the same direction 1542:1559 the same direction as gastric cancer 1542:1577 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 5 30 theme generic 798:804 arg1 method 806:811 the generic method 794:811 the generic method with PNGase F 794:825 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 9 31 theme cancer 1424:1429 arg1 cases 1431:1435 gastric cancer cases 1416:1435 gastric cancer cases 1416:1435 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 1 32 theme glycans 79:85 arg1 study 70:74 a comprehensive study 54:74 a comprehensive study of glycans expressed in biologic systems 54:115 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 1 32 theme glycans 79:85 arg1 Glycomics 43:51 Glycomics 43:51 Glycomics 43:51 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 9 33 with glycans 1328:1334 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 11 34 with controls 1668:1675 arg1 gastritis 1694:1702 nonatrophic gastritis 1682:1702 nonatrophic gastritis 1682:1702 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 0 35 theme gastric 27:33 arg1 cancer 35:40 gastric cancer 27:40 gastric cancer 27:40 Serum glycan signatures of gastric cancer. 24327722 9 36 with glycans 1260:1266 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 5 37 theme serum 774:778 arg1 samples 780:786 serum samples 774:786 serum samples using the generic method with PNGase F 774:825 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 3 38 theme duodenal 385:392 arg1 ulcer 394:398 duodenal ulcer 385:398 duodenal ulcer 385:398 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 9 39 with glycans 1251:1257 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 11 40 theme nonatrophic 1682:1692 arg1 gastritis 1694:1702 nonatrophic gastritis 1682:1702 nonatrophic gastritis 1682:1702 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 10 41 theme serum 1456:1460 arg1 glycans 1462:1468 serum glycans 1456:1468 serum glycans 1456:1468 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 2 42 theme nonatrophic 348:358 arg1 gastritis 360:368 nonatrophic gastritis 348:368 nonatrophic gastritis 348:368 This study aimed to use glycomics to investigate glycan markers that would differentiate patients with gastric cancer from those with nonatrophic gastritis. 24327722 3 43 with infection 494:502 arg1 pylori 522:527 Helicobacter pylori 509:527 Helicobacter pylori 509:527 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 9 44 theme high-mannose-type 1233:1249 arg1 glycans 1251:1257 high-mannose-type glycans 1233:1257 high-mannose-type glycans 1233:1257 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 11 45 theme gastric 1642:1648 arg1 cases 1657:1661 gastric cancer cases 1642:1661 gastric cancer cases from controls with nonatrophic gastritis 1642:1702 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 3 46 theme bacterial 532:540 arg1 infection 542:550 a bacterial infection 530:550 a bacterial infection that can cause either gastric cancer or duodenal ulcer 530:605 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 3 46 theme bacterial 532:540 arg1 infection 494:502 infection 494:502 infection with Helicobacter pylori 494:527 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 1 47 theme sensitive 153:161 arg1 tool 174:177 a simple yet highly sensitive diagnostic tool 133:177 a simple yet highly sensitive diagnostic tool for disease onset and progression 133:211 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 9 48 theme complex 1277:1283 arg1 antenna 1290:1296 one complex type antenna 1273:1296 one complex type antenna 1273:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 4 49 from patients 643:650 arg1 samples 630:636 72 serum samples 621:636 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer 621:742 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 4 49 from patients 643:650 arg1 City 662:665 City 662:665 City 662:665 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 5 50 theme matrix-assisted 848:862 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 9 51 located observed 1404:1411 arg2 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 51 located observed 1404:1411 arg1 cases 1431:1435 gastric cancer cases 1416:1435 gastric cancer cases 1416:1435 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 11 52 theme cancer 1650:1655 arg1 cases 1657:1661 gastric cancer cases 1642:1661 gastric cancer cases from controls with nonatrophic gastritis 1642:1702 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 10 53 theme duodenal 1492:1499 arg1 ulcer 1501:1505 duodenal ulcer 1492:1505 duodenal ulcer 1492:1505 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 5 54 theme PNGase 818:823 arg1 F 825:825 PNGase F 818:825 PNGase F 818:825 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 5 55 theme cyclotron 914:922 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 9 56 theme bigalactosylated 1299:1314 arg1 glycans 1328:1334 bigalactosylated biantennary glycans 1299:1334 bigalactosylated biantennary glycans 1299:1334 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 5 57 theme laser 864:868 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 1 58 theme disease 183:189 arg1 onset 191:195 disease onset 183:195 disease onset 183:195 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 7 59 theme potential 1090:1098 arg1 markers 1100:1106 potential markers 1090:1106 potential markers for each subgroup 1090:1124 ANOVA-based statistical analysis was performed to identify potential markers for each subgroup. 24327722 9 60 theme glycans 1328:1334 arg1 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 3 61 theme gastric 574:580 arg1 cancer 582:587 gastric cancer 574:587 gastric cancer 574:587 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 5 62 theme resonance 924:932 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 6 63 theme corresponding 957:969 arg1 compositions 978:989 The corresponding glycan compositions 953:989 The corresponding glycan compositions 953:989 The corresponding glycan compositions were calculated based on accurate mass. 24327722 12 64 theme protein 1802:1808 arg1 glycosylation 1810:1822 protein glycosylation 1802:1822 protein glycosylation 1802:1822 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 12 65 from role 1794:1797 arg1 pathology 1842:1850 gastric cancer pathology 1827:1850 gastric cancer pathology 1827:1850 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 4 66 theme duodenal 710:717 arg1 ulcer 719:723 duodenal ulcer 710:723 duodenal ulcer 710:723 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 9 67 theme levels 1351:1356 arg1 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 7 68 theme ANOVA-based 1031:1041 arg1 analysis 1055:1062 ANOVA-based statistical analysis 1031:1062 ANOVA-based statistical analysis 1031:1062 ANOVA-based statistical analysis was performed to identify potential markers for each subgroup. 24327722 5 69 theme desorption/ionization 870:890 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 9 70 theme nongalactosylated 1361:1377 arg1 glycans 1391:1397 nongalactosylated biantennary glycans 1361:1397 nongalactosylated biantennary glycans 1361:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 5 71 theme transform-ion 900:912 arg1 spectrometry 939:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry 848:950 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 12 72 gly glycosylation 1810:1822 arg1 cancer 1835:1840 gastric cancer pathology 1827:1850 gastric cancer pathology 1827:1850 Further studies will be needed to validate these findings as biomarkers and identify the role of protein glycosylation in gastric cancer pathology. 24327722 11 73 theme glycan 1586:1591 arg1 profiles 1593:1600 Serum glycan profiles 1580:1600 Serum glycan profiles 1580:1600 Serum glycan profiles may provide biomarkers to differentiate gastric cancer cases from controls with nonatrophic gastritis. 24327722 9 74 theme glycans 1391:1397 arg1 glycans 1260:1266 glycans 1260:1266 glycans with one complex type antenna 1260:1296 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 74 theme glycans 1391:1397 arg1 glycans 1251:1257 high-mannose-type glycans 1233:1257 high-mannose-type glycans 1233:1257 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 74 theme glycans 1391:1397 arg1 glycans 1328:1334 bigalactosylated biantennary glycans 1299:1334 bigalactosylated biantennary glycans 1299:1334 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 74 theme glycans 1391:1397 arg1 levels 1351:1356 increased levels 1341:1356 increased levels of nongalactosylated biantennary glycans 1341:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 9 75 theme decreased 1213:1221 arg1 levels 1223:1228 decreased levels 1213:1228 decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans 1213:1397 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 1 76 theme comprehensive 56:68 arg1 study 70:74 a comprehensive study 54:74 a comprehensive study of glycans expressed in biologic systems 54:115 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 1 76 theme comprehensive 56:68 arg1 Glycomics 43:51 Glycomics 43:51 Glycomics 43:51 Glycomics, a comprehensive study of glycans expressed in biologic systems, is emerging as a simple yet highly sensitive diagnostic tool for disease onset and progression. 24327722 0 77 theme glycan 6:11 arg1 signatures 13:22 Serum glycan signatures 0:22 Serum glycan signatures of gastric cancer 0:40 Serum glycan signatures of gastric cancer. 24327722 9 78 theme gastric 1416:1422 arg1 cases 1431:1435 gastric cancer cases 1416:1435 gastric cancer cases 1416:1435 Generally, decreased levels of high-mannose-type glycans, glycans with one complex type antenna, bigalactosylated biantennary glycans, and increased levels of nongalactosylated biantennary glycans were observed in gastric cancer cases. 24327722 4 79 theme gastric 729:735 arg1 cancer 737:742 gastric cancer 729:742 gastric cancer 729:742 We collected 72 serum samples from patients in Mexico City that presented with nonatrophic gastritis, duodenal ulcer, or gastric cancer. 24327722 3 80 theme Helicobacter 509:520 arg1 pylori 522:527 Helicobacter pylori 509:527 Helicobacter pylori 509:527 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 24327722 0 81 theme cancer 35:40 arg1 signatures 13:22 Serum glycan signatures 0:22 Serum glycan signatures of gastric cancer 0:40 Serum glycan signatures of gastric cancer. 24327722 2 82 with patients 303:310 arg1 cancer 325:330 gastric cancer 317:330 gastric cancer 317:330 This study aimed to use glycomics to investigate glycan markers that would differentiate patients with gastric cancer from those with nonatrophic gastritis. 24327722 10 83 theme gastric 1564:1570 arg1 cancer 1572:1577 gastric cancer 1564:1577 gastric cancer 1564:1577 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 10 84 theme Altered 1438:1444 arg1 levels 1446:1451 Altered levels 1438:1451 Altered levels of serum glycans 1438:1468 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 5 85 attach released 760:767 arg1 samples 780:786 serum samples 774:786 serum samples using the generic method with PNGase F 774:825 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 5 85 attach released 760:767 arg2 N-glycans 745:753 N-glycans 745:753 N-glycans 745:753 N-glycans were released from serum samples using the generic method with PNGase F and were analyzed by matrix-assisted laser desorption/ionization Fourier transform-ion cyclotron resonance mass spectrometry. 24327722 2 86 theme gastric 317:323 arg1 cancer 325:330 gastric cancer 317:330 gastric cancer 317:330 This study aimed to use glycomics to investigate glycan markers that would differentiate patients with gastric cancer from those with nonatrophic gastritis. 24327722 8 87 theme diagnostic 1183:1192 arg1 groups 1194:1199 the diagnostic groups 1179:1199 the diagnostic groups 1179:1199 Nineteen glycans were significantly different among the diagnostic groups. 24327722 10 88 theme glycans 1462:1468 arg1 levels 1446:1451 Altered levels 1438:1451 Altered levels of serum glycans 1438:1468 Altered levels of serum glycans were also observed in duodenal ulcer, but differences were generally in the same direction as gastric cancer. 24327722 3 89 theme different 472:480 arg1 response 482:489 a biologically different response 457:489 a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer 457:605 Patients with duodenal ulcer were also included because they are thought to represent a biologically different response to infection with Helicobacter pylori, a bacterial infection that can cause either gastric cancer or duodenal ulcer. 27223297 7 0 theme Gc 1011:1012 arg1 N794 1014:1017 Gc N794 1011:1017 Gc N794 1011:1017 Results showed that Gc N794, N1035, and N1077 were N-glycosylated but N829 was not. 27223297 4 1 from profile 616:622 arg1 infection 650:658 viral infection 644:658 viral infection via DC-SIGN 644:670 The N-glycosylation profile and significance in viral infection via DC-SIGN have not been elucidated. 27223297 9 2 theme Gc-small 1176:1183 arg1 " 1184:1184 "Gc-small" 1175:1184 "Gc-small" 1175:1184 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 12 3 theme RVFV 1503:1506 arg1 infection 1508:1516 RVFV infection 1503:1516 RVFV infection 1503:1516 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 11 4 theme Gc 1402:1403 arg1 N1077 1405:1409 Gc N1077 1402:1409 Gc N1077 1402:1409 Infection via DC-SIGN was increased in the presence of either Gn N438 or Gc N1077. 27223297 0 5 from N-Glycans 0:8 arg1 Glycoproteins 50:62 the Rift Valley Fever Virus Envelope Glycoproteins 13:62 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 5 from N-Glycans 0:8 arg1 Gc 71:72 Gc 71:72 Gc 71:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 5 from N-Glycans 0:8 arg1 Gn 64:65 Gn 64:65 Gn 64:65 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 3 6 theme -serine 505:511 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 6 theme -serine 505:511 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 7 theme -any 486:489 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 7 theme -any 486:489 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 8 dep sequons 463:469 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 8 dep sequons 463:469 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 9 theme amino 491:495 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 9 theme amino 491:495 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 8 10 theme Gc 1075:1076 arg1 N1077 1078:1082 Gc N1077 1075:1082 Gc N1077 1075:1082 Gc N1077 was heterogeneously N-glycosylated. 27223297 3 11 from positions 535:543 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 11 from positions 535:543 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 11 from positions 535:543 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 11 from positions 535:543 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 11 from positions 535:543 arg1 Gc 591:592 Gc 591:592 Gc 591:592 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 11 from positions 535:543 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 5 12 theme Gc 746:747 arg1 asparagine 749:758 Gc asparagine 746:758 Gc asparagine (N) to glutamine (Q) mutants 746:787 Gc N-glycosylation was first evaluated by using Gc asparagine (N) to glutamine (Q) mutants. 27223297 5 12 theme Gc 746:747 arg1 N 761:761 N 761:761 N 761:761 Gc N-glycosylation was first evaluated by using Gc asparagine (N) to glutamine (Q) mutants. 27223297 6 13 theme strain 852:857 arg1 mutations 888:896 N-to-Q mutations 881:896 N-to-Q mutations 881:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 6 13 theme strain 852:857 arg1 mutants 859:865 recombinant RVFV MP-12 strain mutants 829:865 recombinant RVFV MP-12 strain mutants 829:865 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 3 14 theme N 484:484 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 14 theme N 484:484 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 15 theme virus 398:402 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 1 16 theme mosquito-transmitted 146:165 arg1 fever 135:139 Rift Valley fever 123:139 Rift Valley fever 123:139 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 1 16 theme mosquito-transmitted 146:165 arg1 disease 177:183 a mosquito-transmitted, zoonotic disease 144:183 a mosquito-transmitted, zoonotic disease that infects humans and ruminants 144:217 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 0 17 theme Viral 94:98 arg1 Infection 100:108 Viral Infection 94:108 Viral Infection 94:108 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 8 18 gly N-glycosylated 1104:1117 arg1 N1077 1078:1082 Gc N1077 1075:1082 Gc N1077 1075:1082 Gc N1077 was heterogeneously N-glycosylated. 27223297 3 19 theme Valley 385:390 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 19 theme Valley 385:390 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 6 20 theme Jurkat 937:942 arg1 cells 944:948 Jurkat cells 937:948 Jurkat cells stably expressing DC-SIGN 937:974 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 2 21 theme genus 369:373 arg1 members 342:348 members 342:348 members of the phlebovirus genus 342:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 9 22 dep N-glycoforms 1146:1157 arg1 " 1184:1184 "Gc-small" 1175:1184 "Gc-small" 1175:1184 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 9 22 dep N-glycoforms 1146:1157 arg1 " 1169:1169 "Gc-large" 1160:1169 "Gc-large" 1160:1169 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 6 23 theme N-to-Q 881:886 arg1 mutations 888:896 N-to-Q mutations 881:896 N-to-Q mutations 881:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 6 23 theme N-to-Q 881:886 arg1 mutants 859:865 recombinant RVFV MP-12 strain mutants 829:865 recombinant RVFV MP-12 strain mutants 829:865 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 12 24 theme Gn 1458:1459 arg1 glycoproteins 1469:1481 the Gc and Gn surface glycoproteins 1447:1481 glycoproteins 1469:1481 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 3 25 theme X 503:503 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 25 theme X 503:503 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 2 26 theme phlebovirus 357:367 arg1 genus 369:373 the phlebovirus genus 353:373 the phlebovirus genus 353:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 9 27 theme RVFV 1120:1123 arg1 Gc 1125:1126 RVFV Gc 1120:1126 RVFV Gc 1120:1126 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 11 28 theme N1077 1405:1409 arg1 presence 1372:1379 the presence 1368:1379 the presence of either Gn N438 or Gc N1077 1368:1409 Infection via DC-SIGN was increased in the presence of either Gn N438 or Gc N1077. 27223297 10 29 theme increased 1246:1254 arg1 infection 1256:1264 increased infection 1246:1264 increased infection of cells expressing DC-SIGN 1246:1292 RVFV showed increased infection of cells expressing DC-SIGN compared to cells lacking DC-SIGN. 27223297 2 30 theme non-integrin 296:307 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 30 theme non-integrin 296:307 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 30 theme non-integrin 296:307 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 30 theme non-integrin 296:307 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 3 31 theme S 514:514 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 31 theme S 514:514 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 6 32 theme mutants 859:865 arg1 series 819:824 a series 817:824 a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations 817:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 4 33 theme N-glycosylation 600:614 arg1 profile 616:622 N-glycosylation profile 600:622 N-glycosylation profile 600:622 The N-glycosylation profile and significance in viral infection via DC-SIGN have not been elucidated. 27223297 0 34 dep Glycoproteins 50:62 arg1 Glycoproteins 50:62 the Rift Valley Fever Virus Envelope Glycoproteins 13:62 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 34 dep Glycoproteins 50:62 arg1 Gc 71:72 Gc 71:72 Gc 71:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 34 dep Glycoproteins 50:62 arg1 Gn 64:65 Gn 64:65 Gn 64:65 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 3 35 theme Rift 380:383 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 35 theme Rift 380:383 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 1 36 theme Valley 128:133 arg1 fever 135:139 Rift Valley fever 123:139 Rift Valley fever 123:139 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 1 36 theme Valley 128:133 arg1 disease 177:183 a mosquito-transmitted, zoonotic disease 144:183 a mosquito-transmitted, zoonotic disease that infects humans and ruminants 144:217 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 0 37 theme Fever 29:33 arg1 Virus 35:39 the Rift Valley Fever Virus 13:39 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 5 38 theme asparagine 749:758 arg1 mutants 781:787 Gc asparagine (N) to glutamine (Q) mutants 746:787 Gc asparagine (N) to glutamine (Q) mutants 746:787 Gc N-glycosylation was first evaluated by using Gc asparagine (N) to glutamine (Q) mutants. 27223297 5 38 theme asparagine 749:758 arg1 Q 778:778 Q 778:778 Q 778:778 Gc N-glycosylation was first evaluated by using Gc asparagine (N) to glutamine (Q) mutants. 27223297 9 39 theme " 1226:1226 arg1 band 1228:1231 "Gc-large" band 1217:1231 "Gc-large" band 1217:1231 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 2 40 theme cell 230:233 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 40 theme cell 230:233 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 40 theme cell 230:233 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 40 theme cell 230:233 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 6 41 theme MP-12 846:850 arg1 mutations 888:896 N-to-Q mutations 881:896 N-to-Q mutations 881:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 6 41 theme MP-12 846:850 arg1 mutants 859:865 recombinant RVFV MP-12 strain mutants 829:865 recombinant RVFV MP-12 strain mutants 829:865 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 0 42 theme Valley 22:27 arg1 Virus 35:39 the Rift Valley Fever Virus 13:39 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 9 43 theme distinct 1137:1144 arg1 N-glycoforms 1146:1157 two distinct N-glycoforms 1133:1157 two distinct N-glycoforms: "Gc-large" and "Gc-small" 1133:1184 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 3 44 theme fever 392:396 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 44 theme fever 392:396 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 11 45 theme Gn 1391:1392 arg1 N438 1394:1397 Gn N438 1391:1397 Gn N438 1391:1397 Infection via DC-SIGN was increased in the presence of either Gn N438 or Gc N1077. 27223297 2 46 theme Dendritic 220:228 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 46 theme Dendritic 220:228 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 46 theme Dendritic 220:228 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 46 theme Dendritic 220:228 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 6 47 theme RVFV 841:844 arg1 mutations 888:896 N-to-Q mutations 881:896 N-to-Q mutations 881:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 6 47 theme RVFV 841:844 arg1 mutants 859:865 recombinant RVFV MP-12 strain mutants 829:865 recombinant RVFV MP-12 strain mutants 829:865 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 0 48 theme Envelope 41:48 arg1 Glycoproteins 50:62 the Rift Valley Fever Virus Envelope Glycoproteins 13:62 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 48 theme Envelope 41:48 arg1 Gc 71:72 Gc 71:72 Gc 71:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 48 theme Envelope 41:48 arg1 Gn 64:65 Gn 64:65 Gn 64:65 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 3 49 theme RVFV 405:408 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 49 theme RVFV 405:408 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 1 50 dep mosquito-transmitted 146:165 arg1 zoonotic 168:175 zoonotic 168:175 zoonotic 168:175 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 6 51 from infectivity 907:917 arg1 cells 944:948 Jurkat cells 937:948 Jurkat cells stably expressing DC-SIGN 937:974 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 3 52 theme putative 445:452 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 52 theme putative 445:452 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 0 53 theme Virus 35:39 arg1 Glycoproteins 50:62 the Rift Valley Fever Virus Envelope Glycoproteins 13:62 the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc 13:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 53 theme Virus 35:39 arg1 Gc 71:72 Gc 71:72 Gc 71:72 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 0 53 theme Virus 35:39 arg1 Gn 64:65 Gn 64:65 Gn 64:65 N-Glycans on the Rift Valley Fever Virus Envelope Glycoproteins Gn and Gc Redundantly Support Viral Infection via DC-SIGN. 27223297 10 54 theme cells 1269:1273 arg1 infection 1256:1264 increased infection 1246:1264 increased infection of cells expressing DC-SIGN 1246:1292 RVFV showed increased infection of cells expressing DC-SIGN compared to cells lacking DC-SIGN. 27223297 3 55 gly glycoproteins 411:423 arg1 N1035 573:577 N1035 573:577 N1035 573:577 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 Gn/Gc 426:430 Gn/Gc 426:430 Gn/Gc 426:430 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 N794 561:564 N794 561:564 N794 561:564 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 Gn 552:553 Gn 552:553 Gn 552:553 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 N829 567:570 N829 567:570 N829 567:570 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 55 gly glycoproteins 411:423 arg1 N1077 584:588 N1077 584:588 N1077 (Gc) 584:593 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 9 56 theme Gc-large 1218:1225 arg1 band 1228:1231 "Gc-large" band 1217:1231 "Gc-large" band 1217:1231 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 27223297 3 57 theme N-glycan 454:461 arg1 glycoproteins 411:423 The Rift Valley fever virus (RVFV) glycoproteins 376:423 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) 376:431 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 57 theme N-glycan 454:461 arg1 sequons 463:469 five putative N-glycan sequons 440:469 five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions 440:543 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 12 58 theme surface 1461:1467 arg1 glycoproteins 1469:1481 the Gc and Gn surface glycoproteins 1447:1481 glycoproteins 1469:1481 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 12 59 from N-glycans 1434:1442 arg1 glycoproteins 1469:1481 the Gc and Gn surface glycoproteins 1447:1481 glycoproteins 1469:1481 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 12 59 from N-glycans 1434:1442 arg1 Gc 1451:1452 the Gc and Gn surface glycoproteins 1447:1481 Gc 1451:1452 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 4 60 theme viral 644:648 arg1 infection 650:658 viral infection 644:658 viral infection via DC-SIGN 644:670 The N-glycosylation profile and significance in viral infection via DC-SIGN have not been elucidated. 27223297 3 61 theme acid 497:500 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 61 theme acid 497:500 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 6 62 theme recombinant 829:839 arg1 mutations 888:896 N-to-Q mutations 881:896 N-to-Q mutations 881:896 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 6 62 theme recombinant 829:839 arg1 mutants 859:865 recombinant RVFV MP-12 strain mutants 829:865 recombinant RVFV MP-12 strain mutants 829:865 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 7 63 gly N-glycosylated 1042:1055 arg1 N1077 1031:1035 N1077 1031:1035 N1077 1031:1035 Results showed that Gc N794, N1035, and N1077 were N-glycosylated but N829 was not. 27223297 7 63 gly N-glycosylated 1042:1055 arg1 N794 1014:1017 Gc N794 1011:1017 Gc N794 1011:1017 Results showed that Gc N794, N1035, and N1077 were N-glycosylated but N829 was not. 27223297 7 63 gly N-glycosylated 1042:1055 arg1 N1035 1020:1024 N1035 1020:1024 N1035 1020:1024 Results showed that Gc N794, N1035, and N1077 were N-glycosylated but N829 was not. 27223297 2 64 theme adhesion 258:265 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 64 theme adhesion 258:265 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 64 theme adhesion 258:265 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 64 theme adhesion 258:265 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 1 65 theme Rift 123:126 arg1 fever 135:139 Rift Valley fever 123:139 Rift Valley fever 123:139 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 1 65 theme Rift 123:126 arg1 disease 177:183 a mosquito-transmitted, zoonotic disease 144:183 a mosquito-transmitted, zoonotic disease that infects humans and ruminants 144:217 Rift Valley fever is a mosquito-transmitted, zoonotic disease that infects humans and ruminants. 27223297 6 66 theme mutant 927:932 arg1 infectivity 907:917 the infectivity 903:917 the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN 903:974 Subsequently, we generated a series of recombinant RVFV MP-12 strain mutants, which encode N-to-Q mutations, and the infectivity of each mutant in Jurkat cells stably expressing DC-SIGN was evaluated. 27223297 2 67 theme intercellular 244:256 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 67 theme intercellular 244:256 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 67 theme intercellular 244:256 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 67 theme intercellular 244:256 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 4 68 from significance 628:639 arg1 infection 650:658 viral infection 644:658 viral infection via DC-SIGN 644:670 The N-glycosylation profile and significance in viral infection via DC-SIGN have not been elucidated. 27223297 3 69 theme asparagine 472:481 arg1 T 528:528 T 528:528 T 528:528 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 3 69 theme asparagine 472:481 arg1 /threonine 516:525 asparagine (N)-any amino acid (X)-serine (S)/threonine 472:525 asparagine (N)-any amino acid (X)-serine (S)/threonine (T) 472:529 The Rift Valley fever virus (RVFV) glycoproteins (Gn/Gc) encode five putative N-glycan sequons (asparagine (N)-any amino acid (X)-serine (S)/threonine (T)) at positions: N438 (Gn), and N794, N829, N1035, and N1077 (Gc). 27223297 5 70 theme Gc 698:699 arg1 N-glycosylation 701:715 Gc N-glycosylation 698:715 Gc N-glycosylation 698:715 Gc N-glycosylation was first evaluated by using Gc asparagine (N) to glutamine (Q) mutants. 27223297 12 71 gly glycoproteins 1469:1481 arg1 glycoproteins 1469:1481 the Gc and Gn surface glycoproteins 1447:1481 glycoproteins 1469:1481 Our study showed that N-glycans on the Gc and Gn surface glycoproteins redundantly support RVFV infection via DC-SIGN. 27223297 11 72 theme N438 1394:1397 arg1 presence 1372:1379 the presence 1368:1379 the presence of either Gn N438 or Gc N1077 1368:1409 Infection via DC-SIGN was increased in the presence of either Gn N438 or Gc N1077. 27223297 2 73 theme specific 235:242 arg1 ICAM-3 279:284 ICAM-3 279:284 ICAM-3 279:284 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 73 theme specific 235:242 arg1 DC-SIGN 310:316 DC-SIGN 310:316 DC-SIGN 310:316 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 73 theme specific 235:242 arg1 receptor 329:336 a receptor 327:336 a receptor for members of the phlebovirus genus 327:373 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 2 73 theme specific 235:242 arg1 molecule 267:274 Dendritic cell specific intercellular adhesion molecule 3 220:276 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) 220:317 Dendritic cell specific intercellular adhesion molecule 3 (ICAM-3) grabbing non-integrin (DC-SIGN) acts as a receptor for members of the phlebovirus genus. 27223297 9 74 theme Gc-large 1161:1168 arg1 " 1169:1169 "Gc-large" 1160:1169 "Gc-large" 1160:1169 RVFV Gc made two distinct N-glycoforms: "Gc-large" and "Gc-small", and N1077 was responsible for "Gc-large" band. 28754623 0 0 theme essential 100:108 arg1 production 114:123 essential oil production 100:123 essential oil production of spearmint 100:136 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 3 1 theme quality 754:760 arg1 attributes 762:771 yield and quality attributes 744:771 yield and quality attributes of spearmint (Mentha spicata L.) 744:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 2 2 dep spectroscopy 507:518 arg1 UV-vis 521:526 UV-vis 521:526 UV-vis 521:526 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 1 3 theme radiation-degraded 161:178 arg1 regulators 214:223 regulators 214:223 regulators of growth and development in several crop plants 214:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 3 theme radiation-degraded 161:178 arg1 polysaccharides 180:194 radiation-degraded polysaccharides 161:194 radiation-degraded polysaccharides 161:194 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 5 4 theme spearmint 1193:1201 arg1 oil 1213:1215 spearmint essential oil 1193:1215 spearmint essential oil (EO) 1193:1220 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 5 4 theme spearmint 1193:1201 arg1 EO 1218:1219 EO 1218:1219 EO 1218:1219 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 6 5 theme spectrometry 1335:1346 arg1 analysis 1356:1363 gas chromatography mass spectrometry (GC-MS) analysis 1311:1363 gas chromatography mass spectrometry (GC-MS) analysis 1311:1363 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 5 6 dep content 1172:1178 arg1 the 1168:1170 the 1168:1170 the 1168:1170 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 6 7 theme gas 1311:1313 arg1 GC-MS 1349:1353 GC-MS 1349:1353 GC-MS 1349:1353 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 7 theme gas 1311:1313 arg1 spectrometry 1335:1346 gas chromatography mass spectrometry 1311:1346 gas chromatography mass spectrometry (GC-MS) analysis 1311:1363 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 2 8 theme alginate 381:388 arg1 activity 351:358 previously established growth-promotion activity 311:358 previously established growth-promotion activity of irradiated sodium alginate (ISA) 311:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 6 9 theme carvone 1402:1408 arg1 content 1410:1416 the carvone content 1398:1416 the carvone content 1398:1416 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 5 10 theme essential 1203:1211 arg1 oil 1213:1215 spearmint essential oil 1193:1215 spearmint essential oil (EO) 1193:1220 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 5 10 theme essential 1203:1211 arg1 EO 1218:1219 EO 1218:1219 EO 1218:1219 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 2 11 theme FT-IR 575:579 arg1 studies 582:588 spectroscopic (FT-IR) studies 560:588 spectroscopic (FT-IR) studies 560:588 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 3 12 theme yield 744:748 arg1 attributes 762:771 yield and quality attributes 744:771 yield and quality attributes of spearmint (Mentha spicata L.) 744:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 2 13 theme reasons 300:306 arg1 quest 278:282 quest 278:282 quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA) 278:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 5 14 theme oil 1213:1215 arg1 yield 1184:1188 yield 1184:1188 yield 1184:1188 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 5 14 theme oil 1213:1215 arg1 content 1172:1178 content 1172:1178 content 1172:1178 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 0 15 theme oil 110:112 arg1 production 114:123 essential oil production 100:123 essential oil production of spearmint 100:136 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 3 16 theme spearmint 776:784 arg1 attributes 762:771 yield and quality attributes 744:771 yield and quality attributes of spearmint (Mentha spicata L.) 744:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 2 17 theme relationship 640:651 arg1 understanding 604:616 an understanding 601:616 an understanding of structure-property relationship 601:651 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 5 18 from 80mgL-1 1150:1156 arg1 application 1128:1138 foliar application 1121:1138 foliar application of ISA at 80mgL-1 1121:1156 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 2 19 theme possible 291:298 arg1 reasons 300:306 the possible reasons 287:306 the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA) 287:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 6 20 theme EO 1479:1480 arg1 content 1454:1460 limonene content 1445:1460 limonene content of the spearmint EO 1445:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 2 21 theme activity 351:358 arg1 reasons 300:306 the possible reasons 287:306 the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA) 287:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 0 22 theme spearmint 128:136 arg1 production 114:123 essential oil production 100:123 essential oil production of spearmint 100:136 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 0 22 theme spearmint 128:136 arg1 trichomes 86:94 peltate glandular trichomes 68:94 peltate glandular trichomes 68:94 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 4 23 theme different 813:821 arg1 concentrations 823:836 different concentrations 813:836 different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1] 813:883 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 6 24 theme spearmint 1469:1477 arg1 EO 1479:1480 the spearmint EO 1465:1480 the spearmint EO 1465:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 4 25 dep [0 845:846 arg1 160mgL-1 875:882 160mgL-1 875:882 160mgL-1 875:882 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 4 25 dep [0 845:846 arg1 120 867:869 120 867:869 120 867:869 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 3 26 theme attributes 762:771 arg1 terms 735:739 terms 735:739 terms of yield and quality attributes of spearmint (Mentha spicata L.) 735:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 4 27 theme glandular-trichomes 995:1013 arg1 density 1015:1021 peltate glandular-trichomes density 987:1021 peltate glandular-trichomes density 987:1021 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 6 28 theme % 1393:1393 arg1 decrease 1424:1431 a decrease 1422:1431 a decrease of 15.7% in limonene content of the spearmint EO 1422:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 28 theme % 1393:1393 arg1 increase 1377:1384 an increase 1374:1384 an increase of 18.7% in the carvone content 1374:1416 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 4 29 theme foliar-spray 919:930 arg1 treatment 932:940 the optimum foliar-spray treatment 907:940 the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2 907:1060 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 4 29 theme foliar-spray 919:930 arg1 80mgL-1 886:892 80mgL-1 886:892 80mgL-1 886:892 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 2 30 theme spectroscopic 560:572 arg1 studies 582:588 spectroscopic (FT-IR) studies 560:588 spectroscopic (FT-IR) studies 560:588 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 31 theme growth-promotion 334:349 arg1 activity 351:358 previously established growth-promotion activity 311:358 previously established growth-promotion activity of irradiated sodium alginate (ISA) 311:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 32 theme structural 397:406 arg1 parameters 408:417 structural parameters 397:417 structural parameters of irradiated and un-irradiated sodium alginate 397:465 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 0 33 theme Structural 0:9 arg1 re-arrangement 11:24 Structural re-arrangement 0:24 Structural re-arrangement of depolymerized sodium alginate 0:57 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 5 34 theme ISA 1143:1145 arg1 application 1128:1138 foliar application 1121:1138 foliar application of ISA at 80mgL-1 1121:1156 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 2 35 theme established 322:332 arg1 activity 351:358 previously established growth-promotion activity 311:358 previously established growth-promotion activity of irradiated sodium alginate (ISA) 311:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 0 36 theme depolymerized 29:41 arg1 alginate 50:57 depolymerized sodium alginate 29:57 depolymerized sodium alginate 29:57 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 6 37 theme limonene 1445:1452 arg1 content 1454:1460 limonene content 1445:1460 limonene content of the spearmint EO 1445:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 2 38 theme sodium 451:456 arg1 alginate 458:465 irradiated and un-irradiated sodium alginate 422:465 irradiated and un-irradiated sodium alginate 422:465 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 39 theme alginate 458:465 arg1 parameters 408:417 structural parameters 397:417 structural parameters of irradiated and un-irradiated sodium alginate 397:465 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 3 40 theme Mentha 787:792 arg1 L 802:802 Mentha spicata L. 787:803 Mentha spicata L. 787:803 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 3 40 theme Mentha 787:792 arg1 spearmint 776:784 spearmint 776:784 spearmint (Mentha spicata L.) 776:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 4 41 theme optimum 911:917 arg1 treatment 932:940 the optimum foliar-spray treatment 907:940 the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2 907:1060 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 4 41 theme optimum 911:917 arg1 80mgL-1 886:892 80mgL-1 886:892 80mgL-1 886:892 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 2 42 theme un-irradiated 437:449 arg1 alginate 458:465 irradiated and un-irradiated sodium alginate 422:465 irradiated and un-irradiated sodium alginate 422:465 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 0 43 theme alginate 50:57 arg1 re-arrangement 11:24 Structural re-arrangement 0:24 Structural re-arrangement of depolymerized sodium alginate 0:57 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 3 44 theme spicata 794:800 arg1 L 802:802 Mentha spicata L. 787:803 Mentha spicata L. 787:803 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 3 44 theme spicata 794:800 arg1 spearmint 776:784 spearmint 776:784 spearmint (Mentha spicata L.) 776:804 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 1 45 from plants 267:272 arg1 regulators 214:223 regulators 214:223 regulators of growth and development in several crop plants 214:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 45 from plants 267:272 arg1 polysaccharides 180:194 radiation-degraded polysaccharides 161:194 radiation-degraded polysaccharides 161:194 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 3 46 theme ISA 717:719 arg1 concentrations 699:712 graded concentrations 692:712 graded concentrations of ISA 692:719 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 6 47 from decrease 1424:1431 arg1 content 1410:1416 the carvone content 1398:1416 the carvone content 1398:1416 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 47 from decrease 1424:1431 arg1 content 1454:1460 limonene content 1445:1460 limonene content of the spearmint EO 1445:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 48 theme % 1440:1440 arg1 decrease 1424:1431 a decrease 1422:1431 a decrease of 15.7% in limonene content of the spearmint EO 1422:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 48 theme % 1440:1440 arg1 increase 1377:1384 an increase 1374:1384 an increase of 18.7% in the carvone content 1374:1416 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 1 49 theme growth 228:233 arg1 regulators 214:223 regulators 214:223 regulators of growth and development in several crop plants 214:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 49 theme growth 228:233 arg1 polysaccharides 180:194 radiation-degraded polysaccharides 161:194 radiation-degraded polysaccharides 161:194 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 0 50 theme sodium 43:48 arg1 alginate 50:57 depolymerized sodium alginate 29:57 depolymerized sodium alginate 29:57 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 4 51 theme peltate 987:993 arg1 density 1015:1021 peltate glandular-trichomes density 987:1021 peltate glandular-trichomes density 987:1021 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 3 52 theme graded 692:697 arg1 concentrations 699:712 graded concentrations 692:712 graded concentrations of ISA 692:719 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 3 53 theme foliar 660:665 arg1 application 667:677 foliar application 660:677 foliar application 660:677 Using foliar application, response to graded concentrations of ISA was tested in terms of yield and quality attributes of spearmint (Mentha spicata L.). 28754623 1 54 from growth 228:233 arg1 plants 267:272 several crop plants 254:272 several crop plants 254:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 2 55 theme irradiated 422:431 arg1 alginate 458:465 irradiated and un-irradiated sodium alginate 422:465 irradiated and un-irradiated sodium alginate 422:465 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 0 56 theme peltate 68:74 arg1 trichomes 86:94 peltate glandular trichomes 68:94 peltate glandular trichomes 68:94 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 6 57 theme mass 1330:1333 arg1 GC-MS 1349:1353 GC-MS 1349:1353 GC-MS 1349:1353 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 57 theme mass 1330:1333 arg1 spectrometry 1335:1346 gas chromatography mass spectrometry 1311:1346 gas chromatography mass spectrometry (GC-MS) analysis 1311:1363 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 2 58 theme structure-property 621:638 arg1 relationship 640:651 structure-property relationship 621:651 structure-property relationship 621:651 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 59 dep Transform 541:549 arg1 Infrared 551:558 Infrared 551:558 Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship 541:651 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 60 theme sodium 374:379 arg1 ISA 391:393 ISA 391:393 ISA 391:393 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 60 theme sodium 374:379 arg1 alginate 381:388 irradiated sodium alginate 363:388 irradiated sodium alginate (ISA) 363:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 61 theme Ultraviolet-visible 487:505 arg1 spectroscopy 507:518 Ultraviolet-visible spectroscopy 487:518 Ultraviolet-visible spectroscopy (UV-vis) 487:527 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 1 62 from regulators 214:223 arg1 plants 267:272 several crop plants 254:272 several crop plants 254:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 2 63 theme irradiated 363:372 arg1 ISA 391:393 ISA 391:393 ISA 391:393 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 2 63 theme irradiated 363:372 arg1 alginate 381:388 irradiated sodium alginate 363:388 irradiated sodium alginate (ISA) 363:394 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 1 64 from development 239:249 arg1 plants 267:272 several crop plants 254:272 several crop plants 254:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 65 theme development 239:249 arg1 regulators 214:223 regulators 214:223 regulators of growth and development in several crop plants 214:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 65 theme development 239:249 arg1 polysaccharides 180:194 radiation-degraded polysaccharides 161:194 radiation-degraded polysaccharides 161:194 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 2 66 dep Fourier 533:539 arg1 Transform 541:549 Transform 541:549 Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship 541:651 In quest of the possible reasons of previously established growth-promotion activity of irradiated sodium alginate (ISA), structural parameters of irradiated and un-irradiated sodium alginate were analysed using Ultraviolet-visible spectroscopy (UV-vis) and Fourier Transform Infrared spectroscopic (FT-IR) studies to develop an understanding of structure-property relationship. 28754623 6 67 from increase 1377:1384 arg1 content 1410:1416 the carvone content 1398:1416 the carvone content 1398:1416 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 67 from increase 1377:1384 arg1 content 1454:1460 limonene content 1445:1460 limonene content of the spearmint EO 1445:1480 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 1 68 used used 206:209 arg2 polysaccharides 180:194 radiation-degraded polysaccharides 161:194 radiation-degraded polysaccharides 161:194 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 1 68 used used 206:209 arg2 regulators 214:223 regulators 214:223 regulators of growth and development in several crop plants 214:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 5 69 dep control 1277:1283 arg1 comparison 1259:1268 comparison 1259:1268 comparison 1259:1268 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 1 70 theme past 148:151 arg1 decade 153:158 the past decade 144:158 the past decade 144:158 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 6 71 theme chromatography 1315:1328 arg1 GC-MS 1349:1353 GC-MS 1349:1353 GC-MS 1349:1353 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 6 71 theme chromatography 1315:1328 arg1 spectrometry 1335:1346 gas chromatography mass spectrometry 1311:1346 gas chromatography mass spectrometry (GC-MS) analysis 1311:1363 Compared to the control, gas chromatography mass spectrometry (GC-MS) analysis revealed an increase of 18.7% in the carvone content and a decrease of 15.7% in limonene content of the spearmint EO. 28754623 4 72 theme [0 845:846 arg1 concentrations 823:836 different concentrations 813:836 different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1] 813:883 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 1 73 theme several 254:260 arg1 plants 267:272 several crop plants 254:272 several crop plants 254:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 28754623 0 74 theme glandular 76:84 arg1 trichomes 86:94 peltate glandular trichomes 68:94 peltate glandular trichomes 68:94 Structural re-arrangement of depolymerized sodium alginate enriches peltate glandular trichomes and essential oil production of spearmint. 28754623 5 75 theme foliar 1121:1126 arg1 application 1128:1138 foliar application 1121:1138 foliar application of ISA at 80mgL-1 1121:1156 Measurements made at 150days after planting revealed that foliar application of ISA at 80mgL-1 increased the content and yield of spearmint essential oil (EO) by 36.0 and 122.6%, respectively, in comparison to the control. 28754623 4 76 theme ISA 841:843 arg1 control 849:855 control 849:855 control 849:855 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 4 76 theme ISA 841:843 arg1 [0 845:846 ISA [0 841:846 ISA [0 (control), 40, 80, 120 and 160mgL-1] 841:883 Among different concentrations of ISA [0 (control), 40, 80, 120 and 160mgL-1], 80mgL-1 proved to be the optimum foliar-spray treatment for most of the parameters studied including peltate glandular-trichomes density, which was increased from 20 to 44mm-2. 28754623 1 77 theme crop 262:265 arg1 plants 267:272 several crop plants 254:272 several crop plants 254:272 Over the past decade, radiation-degraded polysaccharides have been used as regulators of growth and development in several crop plants. 26400608 3 0 dep pure 397:400 arg1 homogenous 403:412 homogenous 403:412 homogenous 403:412 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 5 1 theme CS 834:835 arg1 patterns 847:854 CS sulfation patterns 834:854 CS sulfation patterns 834:854 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 2 theme cell 906:909 arg1 types 911:915 different breast cancer cell types 882:915 different breast cancer cell types 882:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 0 3 theme Motifs 87:92 arg1 Synthesis 10:18 Divergent Synthesis 0:18 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 0:136 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 1 4 theme many 173:176 arg1 processes 202:210 many important physiological processes 173:210 many important physiological processes 173:210 Glycosaminoglycans (GAGs) regulate many important physiological processes. 26400608 4 5 theme synthetic 568:576 arg1 strategy 578:585 a versatile synthetic strategy 556:585 a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit 556:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 0 6 theme Triple 107:112 arg1 Cancer 130:135 Triple Negative Breast Cancer 107:135 Triple Negative Breast Cancer 107:135 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 5 7 theme triple 991:996 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 8 theme position 318:325 arg1 groups 344:349 position specific sulfate groups 318:349 position specific sulfate groups 318:349 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 9 theme chondroitin 657:667 arg1 CS 678:679 CS 678:679 CS 678:679 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 4 9 theme chondroitin 657:667 arg1 sulfate 669:675 chondroitin sulfate 657:675 the chondroitin sulfate (CS) repeating unit 653:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 5 10 theme differing 860:868 arg1 effects 870:876 differing effects 860:876 differing effects for different breast cancer cell types 860:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 0 11 theme Breast 123:128 arg1 Cancer 130:135 Triple Negative Breast Cancer 107:135 Triple Negative Breast Cancer 107:135 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 5 12 theme sulfation 837:845 arg1 patterns 847:854 CS sulfation patterns 834:854 CS sulfation patterns 834:854 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 1 13 theme important 178:186 arg1 processes 202:210 many important physiological processes 173:210 many important physiological processes 173:210 Glycosaminoglycans (GAGs) regulate many important physiological processes. 26400608 0 14 theme Negative 114:121 arg1 Cancer 130:135 Triple Negative Breast Cancer 107:135 Triple Negative Breast Cancer 107:135 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 5 15 contain had 856:858 arg1 patterns 847:854 CS sulfation patterns 834:854 CS sulfation patterns 834:854 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 15 contain had 856:858 arg2 effects 870:876 differing effects 860:876 differing effects for different breast cancer cell types 860:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 3 16 theme GAG 414:416 arg1 motifs 418:423 pure, homogenous GAG motifs 397:423 pure, homogenous GAG motifs 397:423 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 1 17 theme physiological 188:200 arg1 processes 202:210 many important physiological processes 173:210 many important physiological processes 173:210 Glycosaminoglycans (GAGs) regulate many important physiological processes. 26400608 5 18 theme Biological 797:806 arg1 evaluation 808:817 Biological evaluation 797:817 Biological evaluation 797:817 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 19 theme inhibitory 935:944 arg1 effect 946:951 the greatest inhibitory effect 922:951 the greatest inhibitory effect 922:951 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 0 20 theme Divergent 0:8 arg1 Synthesis 10:18 Divergent Synthesis 0:18 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 0:136 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 5 21 dep line 1026:1029 arg1 MDA-MB-231 1031:1040 MDA-MB-231 1031:1040 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 22 theme groups 344:349 arg1 introduction 302:313 introduction 302:313 introduction of position specific sulfate groups in the GAG structure 302:370 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 23 theme sulfate 669:675 arg1 unit 692:695 the chondroitin sulfate (CS) repeating unit 653:695 the chondroitin sulfate (CS) repeating unit 653:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 0 24 theme Chondroitin 23:33 arg1 Sulfate 35:41 Chondroitin Sulfate Disaccharides and Identification 23:74 Chondroitin Sulfate Disaccharides and Identification 23:74 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 5 25 theme negative 998:1005 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 26 theme sulfate 336:342 arg1 groups 344:349 position specific sulfate groups 318:349 position specific sulfate groups 318:349 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 27 from patterns 641:648 arg1 unit 692:695 the chondroitin sulfate (CS) repeating unit 653:695 the chondroitin sulfate (CS) repeating unit 653:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 3 28 theme challenging 460:470 arg1 " 453:453 " 453:453 " 453:453 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 3 28 theme challenging 460:470 arg1 task 472:475 a challenging task 458:475 a challenging task 458:475 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 4 29 theme versatile 558:566 arg1 strategy 578:585 a versatile synthetic strategy 556:585 a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit 556:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 2 30 theme specific 327:334 arg1 groups 344:349 position specific sulfate groups 318:349 position specific sulfate groups 318:349 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 31 theme possible 622:629 arg1 patterns 641:648 all the 16 theoretically possible sulfation patterns 597:648 all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit 597:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 5 32 theme cancer 1014:1019 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 33 theme different 882:890 arg1 types 911:915 different breast cancer cell types 882:915 different breast cancer cell types 882:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 0 34 theme Sulfate 35:41 arg1 Synthesis 10:18 Divergent Synthesis 0:18 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 0:136 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 3 35 theme pure 397:400 arg1 motifs 418:423 pure, homogenous GAG motifs 397:423 pure, homogenous GAG motifs 397:423 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 4 36 theme sulfated 743:750 arg1 motifs 752:757 rare but potentially important sulfated motifs 712:757 rare but potentially important sulfated motifs which have not been isolated earlier 712:794 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 3 37 theme motifs 418:423 arg1 procurement 382:392 procurement 382:392 procurement of pure, homogenous GAG motifs to probe the "sulfation 382:447 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 3 38 theme isolation 484:492 arg1 difficulty 494:503 isolation difficulty 484:503 isolation difficulty 484:503 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 0 39 dep Sulfate 35:41 arg1 Disaccharides 43:55 Disaccharides 43:55 Disaccharides 43:55 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 0 39 dep Sulfate 35:41 arg1 Identification 61:74 Identification 61:74 Identification 61:74 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 3 40 theme structural 509:518 arg1 complexity 520:529 structural complexity 509:529 structural complexity 509:529 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 2 41 from introduction 302:313 arg1 structure 362:370 the GAG structure 354:370 the GAG structure 354:370 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 3 42 dep procurement 382:392 arg1 probe 428:432 probe 428:432 to probe the "sulfation 425:447 However, procurement of pure, homogenous GAG motifs to probe the "sulfation code" is a challenging task due to isolation difficulty and structural complexity. 26400608 5 43 theme cell 1021:1024 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 44 theme GAG 358:360 arg1 structure 362:370 the GAG structure 354:370 the GAG structure 354:370 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 45 theme repeating 682:690 arg1 unit 692:695 the chondroitin sulfate (CS) repeating unit 653:695 the chondroitin sulfate (CS) repeating unit 653:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 5 46 theme aggressive 979:988 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 47 theme pertinent 215:223 arg1 issue 225:229 A pertinent issue 213:229 A pertinent issue to address 213:240 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 2 48 theme functional 275:284 arg1 GAGs 253:256 GAGs 253:256 GAGs 253:256 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 2 48 theme functional 275:284 arg1 information 286:296 important functional information 265:296 important functional information 265:296 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 49 theme important 733:741 arg1 motifs 752:757 rare but potentially important sulfated motifs 712:757 rare but potentially important sulfated motifs which have not been isolated earlier 712:794 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 5 50 theme greatest 926:933 arg1 effect 946:951 the greatest inhibitory effect 922:951 the greatest inhibitory effect 922:951 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 4 51 theme rare 712:715 arg1 motifs 752:757 rare but potentially important sulfated motifs 712:757 rare but potentially important sulfated motifs which have not been isolated earlier 712:794 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 0 52 theme Sulfate 79:85 arg1 Motifs 87:92 Sulfate Motifs 79:92 Sulfate Motifs that Inhibit Triple Negative Breast Cancer 79:135 Divergent Synthesis of Chondroitin Sulfate Disaccharides and Identification of Sulfate Motifs that Inhibit Triple Negative Breast Cancer. 26400608 2 53 theme important 265:273 arg1 GAGs 253:256 GAGs 253:256 GAGs 253:256 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 2 53 theme important 265:273 arg1 information 286:296 important functional information 265:296 important functional information 265:296 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26400608 4 54 theme sulfation 631:639 arg1 patterns 641:648 all the 16 theoretically possible sulfation patterns 597:648 all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit 597:695 To this end, we devised a versatile synthetic strategy to obtain all the 16 theoretically possible sulfation patterns in the chondroitin sulfate (CS) repeating unit; these include rare but potentially important sulfated motifs which have not been isolated earlier. 26400608 5 55 theme breast 892:897 arg1 types 911:915 different breast cancer cell types 882:915 different breast cancer cell types 882:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 56 theme breast 1007:1012 arg1 line 1026:1029 triple negative breast cancer cell line 991:1029 triple negative breast cancer cell line MDA-MB-231 991:1040 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 5 57 theme cancer 899:904 arg1 types 911:915 different breast cancer cell types 882:915 different breast cancer cell types 882:915 Biological evaluation indicated that CS sulfation patterns had differing effects for different breast cancer cell types, and the greatest inhibitory effect was observed for the most aggressive, triple negative breast cancer cell line MDA-MB-231. 26400608 2 58 dep issue 225:229 arg1 address 234:240 address 234:240 to address 231:240 A pertinent issue to address is whether GAGs encode important functional information via introduction of position specific sulfate groups in the GAG structure. 26382581 2 0 theme embryonic 553:561 arg1 HEK293rFVII 594:604 HEK293rFVII 594:604 HEK293rFVII 594:604 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 0 theme embryonic 553:561 arg1 cells 587:591 human embryonic kidney-derived (HEK)293 cells 547:591 human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 547:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 1 link kidney-derived 563:576 arg1 HEK293rFVII 594:604 HEK293rFVII 594:604 HEK293rFVII 594:604 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 1 link kidney-derived 563:576 arg1 cells 587:591 human embryonic kidney-derived (HEK)293 cells 547:591 human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 547:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 2 dep cells 718:722 arg1 BHKrFVII 708:715 baby hamster kidney (BHK, BHKrFVII) cells 682:722 BHKrFVII 708:715 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 6 3 theme HEK293rFVII 1383:1393 arg1 glycans 1395:1401 HEK293rFVII glycans 1383:1401 HEK293rFVII glycans 1383:1401 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 3 4 theme HEK293 890:895 arg1 cells 897:901 BHK, CHO, and HEK293 cells 876:901 cells 897:901 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 2 5 theme kidney 695:700 arg1 cells 718:722 baby hamster kidney (BHK, BHKrFVII) cells 682:722 cells 718:722 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 0 6 theme HEK293 102:107 arg1 cells 109:113 BHK, CHO, and HEK293 cells 88:113 cells 109:113 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 2 7 theme baby 682:685 arg1 cells 718:722 baby hamster kidney (BHK, BHKrFVII) cells 682:722 cells 718:722 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 6 8 theme lower 1466:1470 arg1 degree 1472:1477 a lower degree 1464:1477 a lower degree of terminal sialylation 1464:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 2 9 theme rFVII 526:530 arg1 properties 512:521 the biochemical properties 496:521 the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 496:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 6 10 theme galactosamines 1543:1556 arg1 amount 1515:1520 a high amount 1508:1520 a high amount of terminal N-acetyl galactosamines (GalNAc) 1508:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 10 theme galactosamines 1543:1556 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 10 theme galactosamines 1543:1556 arg1 variety 1452:1458 a higher structural variety 1432:1458 a higher structural variety 1432:1458 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 10 theme galactosamines 1543:1556 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 10 theme galactosamines 1543:1556 arg1 degree 1472:1477 a lower degree 1464:1477 a lower degree of terminal sialylation 1464:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 10 theme galactosamines 1543:1556 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 4 11 theme analytical 1059:1068 arg1 results 1070:1076 The analytical results 1055:1076 The analytical results 1055:1076 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 8 12 from cells 1826:1830 arg1 option 1870:1875 the best option 1861:1875 the best option for the production of rFVII 1861:1903 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 8 12 from cells 1826:1830 arg1 CONCLUSIONS 1691:1701 CONCLUSIONS 1691:1701 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells 1691:1830 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 6 13 theme terminal 1525:1532 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 13 theme terminal 1525:1532 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 8 14 theme sialylation 1782:1792 arg1 degree 1772:1777 the highest degree 1760:1777 the highest degree of sialylation and no terminal GalNAc 1760:1815 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 6 15 theme structural 1441:1450 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 15 theme structural 1441:1450 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 15 theme structural 1441:1450 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 15 theme structural 1441:1450 arg1 variety 1452:1458 a higher structural variety 1432:1458 a higher structural variety 1432:1458 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 1 16 theme UNLABELLED 116:125 arg1 rFVII 172:176 rFVII 172:176 rFVII 172:176 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 16 theme UNLABELLED 116:125 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 16 theme UNLABELLED 116:125 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 2 17 theme close 455:459 arg1 modifications 438:450 post-translational modifications 419:450 post-translational modifications as close 419:459 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 0 18 theme VII 71:73 arg1 N-glycosylation 15:29 N-glycosylation 15:29 N-glycosylation of recombinant human coagulation factor VII 15:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 1 19 theme post 276:279 arg1 modifications 295:307 complex post translational modifications 268:307 complex post translational modifications 268:307 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 20 theme METHODS 140:146 arg1 rFVII 172:176 rFVII 172:176 rFVII 172:176 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 20 theme METHODS 140:146 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 20 theme METHODS 140:146 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 2 21 theme hamster 648:654 arg1 ovary 656:660 Chinese hamster ovary 640:660 Chinese hamster ovary (CHO, CHOrFVII) 640:676 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 21 theme hamster 648:654 arg1 CHO 663:665 CHO 663:665 CHO 663:665 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 8 22 theme GalNAc 1810:1815 arg1 degree 1772:1777 the highest degree 1760:1777 the highest degree of sialylation and no terminal GalNAc 1760:1815 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 6 23 theme high 1510:1513 arg1 amount 1515:1520 a high amount 1508:1520 a high amount of terminal N-acetyl galactosamines (GalNAc) 1508:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 23 theme high 1510:1513 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 23 theme high 1510:1513 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 23 theme high 1510:1513 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 1 24 theme factor 160:165 arg1 rFVII 172:176 rFVII 172:176 rFVII 172:176 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 24 theme factor 160:165 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 24 theme factor 160:165 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 0 25 from Differences 0:10 arg1 N-glycosylation 15:29 N-glycosylation 15:29 N-glycosylation of recombinant human coagulation factor VII 15:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 5 26 theme protein 1352:1358 arg1 features 1360:1367 all other protein features 1342:1367 all other protein features 1342:1367 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 2 27 dep rFVII 408:412 arg1 compared 487:494 compared 487:494 compared 487:494 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 4 28 theme FVII 1135:1138 arg1 proteins 1140:1147 the various FVII proteins 1123:1147 the various FVII proteins 1123:1147 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 1 29 theme mammalian 322:330 arg1 cells 332:336 mammalian cells 322:336 mammalian cells 322:336 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 8 30 theme best 1865:1868 arg1 option 1870:1875 the best option 1861:1875 the best option for the production of rFVII 1861:1903 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 8 30 theme best 1865:1868 arg1 CONCLUSIONS 1691:1701 CONCLUSIONS 1691:1701 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells 1691:1830 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 6 31 theme sialylation 1491:1501 arg1 amount 1515:1520 a high amount 1508:1520 a high amount of terminal N-acetyl galactosamines (GalNAc) 1508:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 31 theme sialylation 1491:1501 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 31 theme sialylation 1491:1501 arg1 variety 1452:1458 a higher structural variety 1432:1458 a higher structural variety 1432:1458 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 31 theme sialylation 1491:1501 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 31 theme sialylation 1491:1501 arg1 degree 1472:1477 a lower degree 1464:1477 a lower degree of terminal sialylation 1464:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 31 theme sialylation 1491:1501 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 2 32 dep CHO 663:665 arg1 CHOrFVII 668:675 CHOrFVII 668:675 CHOrFVII 668:675 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 4 33 theme apparent 1088:1095 arg1 differences 1103:1113 no apparent gross differences 1085:1113 no apparent gross differences between the various FVII proteins 1085:1147 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 5 34 theme HEK293 1240:1245 arg1 cells 1247:1251 HEK293 cells 1240:1251 HEK293 cells 1240:1251 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 2 35 dep derived 755:761 arg1 plasma 748:753 plasma 748:753 plasma 748:753 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 36 theme line 383:386 arg1 suitability 355:365 the suitability 351:365 the suitability of a human cell line 351:386 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 37 theme human 372:376 arg1 line 383:386 a human cell line 370:386 a human cell line 370:386 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 4 38 theme N-linked 1166:1173 arg1 pattern 1189:1195 their N-linked glycosylation pattern 1160:1195 their N-linked glycosylation pattern 1160:1195 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 0 39 theme recombinant 34:44 arg1 VII 71:73 recombinant human coagulation factor VII 34:73 recombinant human coagulation factor VII 34:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 3 40 theme protein 967:973 arg1 activity 975:982 protein activity 967:982 protein activity 967:982 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 0 41 theme coagulation 52:62 arg1 VII 71:73 recombinant human coagulation factor VII 34:73 recombinant human coagulation factor VII 34:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 7 42 contain contained 1601:1609 arg2 structures 1679:1688 hybrid and high mannose (Man) structures 1649:1688 hybrid and high mannose (Man) structures 1649:1688 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 7 42 contain contained 1601:1609 arg2 fucoses 1623:1629 one or more fucoses 1611:1629 one or more fucoses (Fuc) 1611:1635 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 7 42 contain contained 1601:1609 arg1 oligosaccharides 1584:1599 All HEK293rFVII oligosaccharides 1568:1599 All HEK293rFVII oligosaccharides 1568:1599 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 7 42 contain contained 1601:1609 arg2 Fuc 1632:1634 Fuc 1632:1634 Fuc 1632:1634 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 2 43 theme analytical 792:801 arg1 methods 803:809 various analytical methods 784:809 various analytical methods 784:809 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 5 44 theme CHO 1285:1287 arg1 cells 1297:1301 CHO and BHK cells 1285:1301 cells 1297:1301 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 7 45 theme high 1660:1663 arg1 Man 1674:1676 Man 1674:1676 Man 1674:1676 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 7 45 theme high 1660:1663 arg1 mannose 1665:1671 high mannose 1660:1671 high mannose (Man) 1660:1677 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 1 46 theme activated 219:227 arg1 rFVIIa 235:240 rFVIIa 235:240 rFVIIa 235:240 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 46 theme activated 219:227 arg1 FVII 229:232 recombinant activated FVII 207:232 recombinant activated FVII (rFVIIa) 207:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 7 47 theme hybrid 1649:1654 arg1 structures 1679:1688 hybrid and high mannose (Man) structures 1649:1688 hybrid and high mannose (Man) structures 1649:1688 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 2 48 theme human 547:551 arg1 HEK293rFVII 594:604 HEK293rFVII 594:604 HEK293rFVII 594:604 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 48 theme human 547:551 arg1 cells 587:591 human embryonic kidney-derived (HEK)293 cells 547:591 human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 547:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 3 49 theme BHK 876:878 arg1 CHO 881:883 BHK, CHO, and HEK293 cells 876:901 CHO 881:883 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 2 50 theme derived 755:761 arg1 FVII 763:766 plasma derived FVII 748:766 plasma derived FVII (pdFVII) 748:775 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 50 theme derived 755:761 arg1 pdFVII 769:774 pdFVII 769:774 pdFVII 769:774 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 51 theme HEK 579:581 arg1 HEK293rFVII 594:604 HEK293rFVII 594:604 HEK293rFVII 594:604 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 51 theme HEK 579:581 arg1 cells 587:591 human embryonic kidney-derived (HEK)293 cells 547:591 human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 547:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 5 52 theme BHK 1293:1295 arg1 cells 1297:1301 CHO and BHK cells 1285:1301 cells 1297:1301 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 2 53 theme kidney-derived 563:576 arg1 HEK293rFVII 594:604 HEK293rFVII 594:604 HEK293rFVII 594:604 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 53 theme kidney-derived 563:576 arg1 cells 587:591 human embryonic kidney-derived (HEK)293 cells 547:591 human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 547:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 8 54 contain contained 1750:1758 arg1 CHOrFVII 1741:1748 CHOrFVII 1741:1748 CHOrFVII 1741:1748 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 8 54 contain contained 1750:1758 arg2 degree 1772:1777 the highest degree 1760:1777 the highest degree of sialylation and no terminal GalNAc 1760:1815 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 2 55 theme hamster 687:693 arg1 cells 718:722 baby hamster kidney (BHK, BHKrFVII) cells 682:722 cells 718:722 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 8 56 theme highest 1764:1770 arg1 degree 1772:1777 the highest degree 1760:1777 the highest degree of sialylation and no terminal GalNAc 1760:1815 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 1 57 theme precursor 184:192 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 57 theme precursor 184:192 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 2 58 theme Chinese 640:646 arg1 ovary 656:660 Chinese hamster ovary 640:660 Chinese hamster ovary (CHO, CHOrFVII) 640:676 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 58 theme Chinese 640:646 arg1 CHO 663:665 CHO 663:665 CHO 663:665 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 6 59 theme N-acetyl 1534:1541 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 59 theme N-acetyl 1534:1541 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 2 60 theme biochemical 500:510 arg1 properties 512:521 the biochemical properties 496:521 the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) 496:605 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 8 61 theme terminal 1801:1808 arg1 GalNAc 1810:1815 no terminal GalNAc 1798:1815 no terminal GalNAc 1798:1815 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 6 62 theme higher 1434:1439 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 62 theme higher 1434:1439 arg1 GalNAc 1559:1564 GalNAc 1559:1564 GalNAc 1559:1564 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 62 theme higher 1434:1439 arg1 galactosamines 1543:1556 terminal N-acetyl galactosamines 1525:1556 terminal N-acetyl galactosamines (GalNAc) 1525:1565 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 6 62 theme higher 1434:1439 arg1 variety 1452:1458 a higher structural variety 1432:1458 a higher structural variety 1432:1458 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 1 63 theme BACKGROUND & 127:138 arg1 rFVII 172:176 rFVII 172:176 rFVII 172:176 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 63 theme BACKGROUND & 127:138 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 63 theme BACKGROUND & 127:138 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 0 64 theme BHK 88:90 arg1 CHO 93:95 BHK, CHO, and HEK293 cells 88:113 CHO 93:95 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 1 65 theme translational 281:293 arg1 modifications 295:307 complex post translational modifications 268:307 complex post translational modifications 268:307 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 2 66 with rFVII 408:412 arg1 modifications 438:450 post-translational modifications 419:450 post-translational modifications as close 419:459 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 1 67 theme Recombinant 148:158 arg1 rFVII 172:176 rFVII 172:176 rFVII 172:176 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 67 theme Recombinant 148:158 arg1 VII 167:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII 116:169 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII) 116:177 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 67 theme Recombinant 148:158 arg1 molecule 194:201 the precursor molecule 180:201 the precursor molecule for recombinant activated FVII (rFVIIa) 180:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 2 68 theme post-translational 419:436 arg1 modifications 438:450 post-translational modifications 419:450 post-translational modifications as close 419:459 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 5 69 theme other 1346:1350 arg1 features 1360:1367 all other protein features 1342:1367 all other protein features 1342:1367 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 4 70 theme various 1127:1133 arg1 proteins 1140:1147 the various FVII proteins 1123:1147 the various FVII proteins 1123:1147 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 8 71 theme CHO 1822:1824 arg1 cells 1826:1830 CHO cells 1822:1830 CHO cells 1822:1830 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 3 72 theme stable 909:914 arg1 transfection 916:927 stable transfection 909:927 stable transfection 909:927 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 4 73 theme gross 1097:1101 arg1 differences 1103:1113 no apparent gross differences 1085:1113 no apparent gross differences between the various FVII proteins 1085:1147 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 3 74 theme rFVII 934:938 arg1 isolates 940:947 rFVII isolates 934:947 rFVII isolates 934:947 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 6 75 theme terminal 1482:1489 arg1 sialylation 1491:1501 terminal sialylation 1482:1501 terminal sialylation 1482:1501 HEK293rFVII glycans were mainly characterized by a higher structural variety and a lower degree of terminal sialylation, and a high amount of terminal N-acetyl galactosamines (GalNAc). 26382581 8 76 theme rFVII 1899:1903 arg1 production 1885:1894 the production 1881:1894 the production of rFVII 1881:1903 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 2 77 theme various 784:790 arg1 methods 803:809 various analytical methods 784:809 various analytical methods 784:809 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 5 78 theme Most 1198:1201 arg1 N-glycans 1203:1211 Most N-glycans 1198:1211 Most N-glycans found on rFVII produced in HEK293 cells 1198:1251 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 2 79 theme cell 378:381 arg1 line 383:386 a human cell line 370:386 a human cell line 370:386 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 4 80 theme glycosylation 1175:1187 arg1 pattern 1189:1195 their N-linked glycosylation pattern 1160:1195 their N-linked glycosylation pattern 1160:1195 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 26382581 0 81 theme human 46:50 arg1 VII 71:73 recombinant human coagulation factor VII 34:73 recombinant human coagulation factor VII 34:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 0 82 theme factor 64:69 arg1 VII 71:73 recombinant human coagulation factor VII 34:73 recombinant human coagulation factor VII 34:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 3 83 theme post-translational 1000:1017 arg1 modifications 1019:1031 post-translational modifications 1000:1031 post-translational modifications 1000:1031 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 0 84 gly N-glycosylation 15:29 arg1 VII 71:73 recombinant human coagulation factor VII 34:73 recombinant human coagulation factor VII 34:73 Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. 26382581 3 85 theme selected 836:843 arg1 clones 856:861 selected production clones 836:861 selected production clones derived from BHK, CHO, and HEK293 cells 836:901 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 5 86 from cells 1297:1301 arg1 rFVII 1274:1278 rFVII 1274:1278 rFVII from CHO and BHK cells 1274:1301 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 8 87 from isolates 1718:1725 arg1 option 1870:1875 the best option 1861:1875 the best option for the production of rFVII 1861:1903 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 8 87 from isolates 1718:1725 arg1 CONCLUSIONS 1691:1701 CONCLUSIONS 1691:1701 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells 1691:1830 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 3 88 attach derived 863:869 arg1 CHO 881:883 BHK, CHO, and HEK293 cells 876:901 CHO 881:883 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 3 88 attach derived 863:869 arg2 clones 856:861 selected production clones 836:861 selected production clones derived from BHK, CHO, and HEK293 cells 836:901 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 3 88 attach derived 863:869 arg1 cells 897:901 BHK, CHO, and HEK293 cells 876:901 cells 897:901 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 1 89 theme complex 268:274 arg1 modifications 295:307 complex post translational modifications 268:307 complex post translational modifications 268:307 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 3 90 theme production 845:854 arg1 clones 856:861 selected production clones 836:861 selected production clones derived from BHK, CHO, and HEK293 cells 836:901 rFVII was purified from selected production clones derived from BHK, CHO, and HEK293 cells after stable transfection, and rFVII isolates were analyzed for protein activity, impurities and post-translational modifications. 26382581 5 91 located found 1213:1217 arg2 N-glycans 1203:1211 Most N-glycans 1198:1211 Most N-glycans found on rFVII produced in HEK293 cells 1198:1251 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 5 91 located found 1213:1217 arg1 rFVII 1222:1226 rFVII 1222:1226 rFVII produced in HEK293 cells 1222:1251 Most N-glycans found on rFVII produced in HEK293 cells were not detected on rFVII from CHO and BHK cells, or, somewhat unexpectedly, on pdFVII; all other protein features were similar. 26382581 2 92 link derived 755:761 arg1 FVII 763:766 plasma derived FVII 748:766 plasma derived FVII (pdFVII) 748:775 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 2 92 link derived 755:761 arg1 pdFVII 769:774 pdFVII 769:774 pdFVII 769:774 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 1 93 theme recombinant 207:217 arg1 rFVIIa 235:240 rFVIIa 235:240 rFVIIa 235:240 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 1 93 theme recombinant 207:217 arg1 FVII 229:232 recombinant activated FVII 207:232 recombinant activated FVII (rFVIIa) 207:241 UNLABELLED BACKGROUND & METHODS Recombinant factor VII (rFVII), the precursor molecule for recombinant activated FVII (rFVIIa), is, due to its need for complex post translational modifications, produced in mammalian cells. 26382581 7 94 theme mannose 1665:1671 arg1 structures 1679:1688 hybrid and high mannose (Man) structures 1649:1688 hybrid and high mannose (Man) structures 1649:1688 All HEK293rFVII oligosaccharides contained one or more fucoses (Fuc), as well as hybrid and high mannose (Man) structures. 26382581 2 95 theme BHK 703:705 arg1 cells 718:722 baby hamster kidney (BHK, BHKrFVII) cells 682:722 cells 718:722 To evaluate the suitability of a human cell line in order to produce rFVII with post-translational modifications as close as possible to pdFVII, we compared the biochemical properties of rFVII synthesized in human embryonic kidney-derived (HEK)293 cells (HEK293rFVII) with those of rFVII expressed in Chinese hamster ovary (CHO, CHOrFVII) and baby hamster kidney (BHK, BHKrFVII) cells, and also with those of plasma derived FVII (pdFVII), using various analytical methods. 26382581 8 96 from contained 1750:1758 arg1 option 1870:1875 the best option 1861:1875 the best option for the production of rFVII 1861:1903 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 8 96 from contained 1750:1758 arg1 CONCLUSIONS 1691:1701 CONCLUSIONS 1691:1701 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells 1691:1830 CONCLUSIONS From all rFVII isolates investigated, CHOrFVII contained the highest degree of sialylation and no terminal GalNAc, and CHO cells were therefore assumed to be the best option for the production of rFVII. 26382581 4 97 link N-linked 1166:1173 arg1 pattern 1189:1195 their N-linked glycosylation pattern 1160:1195 their N-linked glycosylation pattern 1160:1195 RESULTS & DISCUSSION The analytical results showed no apparent gross differences between the various FVII proteins, except in their N-linked glycosylation pattern. 28917901 6 0 theme new 888:890 arg1 tissue 908:913 the new tissue was bone tissue 884:913 the new tissue was bone tissue 884:913 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 7 1 theme tissue 1063:1068 arg1 applications 1082:1093 bone tissue engineering applications 1058:1093 bone tissue engineering applications 1058:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 6 2 theme cranial 710:716 arg1 repair 723:728 rat cranial bone repair 706:728 rat cranial bone repair 706:728 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 1 3 theme bone 237:240 arg1 regeneration 242:253 the complicated cranial bone regeneration 213:253 the complicated cranial bone regeneration 213:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 5 4 theme crosslinking 629:640 arg1 methods 642:648 crosslinking methods 629:648 crosslinking methods 629:648 The degradation behavior of the hydrogel is influenced by hydrogel composition, crosslinking methods and degradation environment. 28917901 6 5 theme " 796:796 arg1 tissue 802:807 "white" new tissue 790:807 "white" new tissue 790:807 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 2 6 theme composite 355:363 arg1 bio-glass 365:373 composite bio-glass 355:373 composite bio-glass (BG) 355:378 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 2 6 theme composite 355:363 arg1 BG 376:377 BG 376:377 BG 376:377 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 6 7 theme white 791:795 arg1 tissue 802:807 "white" new tissue 790:807 "white" new tissue 790:807 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 2 8 theme novel 285:289 arg1 polysaccharides 310:324 a novel triple crosslinked polysaccharides 283:324 a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair 283:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 7 9 link crosslinked 1020:1030 arg1 potential 1044:1052 potential 1044:1052 potential for bone tissue engineering applications 1044:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 7 9 link crosslinked 1020:1030 arg1 hydrogel 1032:1039 the prepared triple crosslinked hydrogel 1000:1039 the prepared triple crosslinked hydrogel 1000:1039 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 7 10 theme bone 1058:1061 arg1 applications 1082:1093 bone tissue engineering applications 1058:1093 bone tissue engineering applications 1058:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 5 11 theme hydrogel 581:588 arg1 behavior 565:572 The degradation behavior 549:572 The degradation behavior of the hydrogel 549:588 The degradation behavior of the hydrogel is influenced by hydrogel composition, crosslinking methods and degradation environment. 28917901 6 12 theme rat 706:708 arg1 repair 723:728 rat cranial bone repair 706:728 rat cranial bone repair 706:728 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 6 13 theme histopathological 847:863 arg1 analysis 865:872 HE histopathological analysis 844:872 HE histopathological analysis 844:872 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 2 14 theme bone 392:395 arg1 repair 397:402 cranial bone repair 384:402 cranial bone repair 384:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 6 15 theme HE 844:845 arg1 analysis 865:872 HE histopathological analysis 844:872 HE histopathological analysis 844:872 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 1 16 theme Bone 89:92 arg1 problem 121:127 a public health problem 105:127 a public health problem around the word 105:143 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 1 16 theme Bone 89:92 arg1 disease 94:100 Bone disease 89:100 Bone disease 89:100 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 2 17 theme cranial 384:390 arg1 repair 397:402 cranial bone repair 384:402 cranial bone repair 384:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 5 18 theme hydrogel 607:614 arg1 composition 616:626 hydrogel composition 607:626 hydrogel composition 607:626 The degradation behavior of the hydrogel is influenced by hydrogel composition, crosslinking methods and degradation environment. 28917901 5 19 theme degradation 553:563 arg1 behavior 565:572 The degradation behavior 549:572 The degradation behavior of the hydrogel 549:588 The degradation behavior of the hydrogel is influenced by hydrogel composition, crosslinking methods and degradation environment. 28917901 2 20 theme crosslinked 298:308 arg1 polysaccharides 310:324 a novel triple crosslinked polysaccharides 283:324 a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair 283:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 7 21 theme engineering 1070:1080 arg1 applications 1082:1093 bone tissue engineering applications 1058:1093 bone tissue engineering applications 1058:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 0 22 theme hydrogel 33:40 arg1 bio-glass 54:62 injectable hydrogel compositing bio-glass 22:62 injectable hydrogel compositing bio-glass for cranial bone repair 22:86 Polysaccharides based injectable hydrogel compositing bio-glass for cranial bone repair. 28917901 7 23 theme prepared 1004:1011 arg1 potential 1044:1052 potential 1044:1052 potential for bone tissue engineering applications 1044:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 7 23 theme prepared 1004:1011 arg1 hydrogel 1032:1039 the prepared triple crosslinked hydrogel 1000:1039 the prepared triple crosslinked hydrogel 1000:1039 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 2 24 theme triple 291:296 arg1 polysaccharides 310:324 a novel triple crosslinked polysaccharides 283:324 a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair 283:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 1 25 theme novel 174:178 arg1 scaffolds 199:207 novel tissue engineering scaffolds 174:207 novel tissue engineering scaffolds for the complicated cranial bone regeneration 174:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 0 26 theme injectable 22:31 arg1 bio-glass 54:62 injectable hydrogel compositing bio-glass 22:62 injectable hydrogel compositing bio-glass for cranial bone repair 22:86 Polysaccharides based injectable hydrogel compositing bio-glass for cranial bone repair. 28917901 7 27 theme triple 1013:1018 arg1 potential 1044:1052 potential 1044:1052 potential for bone tissue engineering applications 1044:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 7 27 theme triple 1013:1018 arg1 hydrogel 1032:1039 the prepared triple crosslinked hydrogel 1000:1039 the prepared triple crosslinked hydrogel 1000:1039 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 1 28 theme public 107:112 arg1 problem 121:127 a public health problem 105:127 a public health problem around the word 105:143 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 1 28 theme public 107:112 arg1 disease 94:100 Bone disease 89:100 Bone disease 89:100 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 1 29 theme tissue 180:185 arg1 scaffolds 199:207 novel tissue engineering scaffolds 174:207 novel tissue engineering scaffolds for the complicated cranial bone regeneration 174:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 3 30 theme Dynamic 405:411 arg1 analysis 424:431 Dynamic mechanical analysis 405:431 Dynamic mechanical analysis 405:431 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 6 31 located detected 813:820 arg2 chunks 780:785 chunks 780:785 chunks of "white" new tissue 780:807 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 6 31 located detected 813:820 arg1 site 838:841 the defected site 825:841 the defected site 825:841 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 6 32 theme defected 829:836 arg1 site 838:841 the defected site 825:841 the defected site 825:841 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 1 33 theme engineering 187:197 arg1 scaffolds 199:207 novel tissue engineering scaffolds 174:207 novel tissue engineering scaffolds for the complicated cranial bone regeneration 174:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 0 34 theme compositing 42:52 arg1 bio-glass 54:62 injectable hydrogel compositing bio-glass 22:62 injectable hydrogel compositing bio-glass for cranial bone repair 22:86 Polysaccharides based injectable hydrogel compositing bio-glass for cranial bone repair. 28917901 4 35 theme compositing 513:523 arg1 BG 525:526 compositing BG 513:526 compositing BG 513:526 While after compositing BG, G' exceeded 4500Pa. 28917901 3 36 theme mechanical 413:422 arg1 analysis 424:431 Dynamic mechanical analysis 405:431 Dynamic mechanical analysis 405:431 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 7 37 theme bone 973:976 arg1 repair 978:983 bone repair 973:983 bone repair 973:983 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 6 38 theme was 899:901 arg1 tissue 908:913 the new tissue was bone tissue 884:913 the new tissue was bone tissue 884:913 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 0 39 theme cranial 68:74 arg1 repair 81:86 cranial bone repair 68:86 cranial bone repair 68:86 Polysaccharides based injectable hydrogel compositing bio-glass for cranial bone repair. 28917901 5 40 theme degradation 654:664 arg1 environment 666:676 degradation environment 654:676 degradation environment 654:676 The degradation behavior of the hydrogel is influenced by hydrogel composition, crosslinking methods and degradation environment. 28917901 7 41 theme crosslinked 1020:1030 arg1 potential 1044:1052 potential 1044:1052 potential for bone tissue engineering applications 1044:1093 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 7 41 theme crosslinked 1020:1030 arg1 hydrogel 1032:1039 the prepared triple crosslinked hydrogel 1000:1039 the prepared triple crosslinked hydrogel 1000:1039 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 6 42 theme regeneration 746:757 arg1 effect 759:764 excellent bone regeneration effect 731:764 excellent bone regeneration effect 731:764 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 1 43 theme health 114:119 arg1 problem 121:127 a public health problem 105:127 a public health problem around the word 105:143 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 1 43 theme health 114:119 arg1 disease 94:100 Bone disease 89:100 Bone disease 89:100 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 6 44 theme bone 903:906 arg1 tissue 908:913 the new tissue was bone tissue 884:913 the new tissue was bone tissue 884:913 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 6 45 theme bone 741:744 arg1 regeneration 746:757 excellent bone regeneration 731:757 excellent bone regeneration effect 731:764 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 3 46 theme hydrogel 472:479 arg1 G 461:461 G' 461:462 G' 461:462 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 3 46 theme hydrogel 472:479 arg1 modulus 452:458 the storage modulus 440:458 the storage modulus (G') of the hydrogel reached to ∼4000Pa 440:498 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 2 47 link crosslinked 298:308 arg1 polysaccharides 310:324 a novel triple crosslinked polysaccharides 283:324 a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair 283:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 6 48 theme excellent 731:739 arg1 regeneration 746:757 excellent bone regeneration 731:757 excellent bone regeneration effect 731:764 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 2 49 theme injectable 332:341 arg1 hydrogel 343:350 injectable hydrogel 332:350 injectable hydrogel to composite bio-glass (BG) for cranial bone repair 332:402 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 2 50 theme present 260:266 arg1 work 268:271 The present work 256:271 The present work 256:271 The present work developed a novel triple crosslinked polysaccharides based injectable hydrogel to composite bio-glass (BG) for cranial bone repair. 28917901 6 51 theme tissue 802:807 arg1 chunks 780:785 chunks 780:785 chunks of "white" new tissue 780:807 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 7 52 theme BG 966:967 arg1 carrier 955:961 the carrier 951:961 the carrier of BG for bone repair 951:983 Thus, the hydrogel is suitable as the carrier of BG for bone repair, demonstrating the prepared triple crosslinked hydrogel is potential for bone tissue engineering applications. 28917901 6 53 theme new 798:800 arg1 tissue 802:807 "white" new tissue 790:807 "white" new tissue 790:807 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 3 54 theme storage 444:450 arg1 G 461:461 G' 461:462 G' 461:462 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 3 54 theme storage 444:450 arg1 modulus 452:458 the storage modulus 440:458 the storage modulus (G') of the hydrogel reached to ∼4000Pa 440:498 Dynamic mechanical analysis showed the storage modulus (G') of the hydrogel reached to ∼4000Pa. 28917901 1 55 theme complicated 217:227 arg1 regeneration 242:253 the complicated cranial bone regeneration 213:253 the complicated cranial bone regeneration 213:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 28917901 0 56 theme bone 76:79 arg1 repair 81:86 cranial bone repair 68:86 cranial bone repair 68:86 Polysaccharides based injectable hydrogel compositing bio-glass for cranial bone repair. 28917901 6 57 theme tissue 892:897 arg1 tissue 908:913 the new tissue was bone tissue 884:913 the new tissue was bone tissue 884:913 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 6 58 theme bone 718:721 arg1 repair 723:728 rat cranial bone repair 706:728 rat cranial bone repair 706:728 Through compositing BG for rat cranial bone repair, excellent bone regeneration effect was achieved (chunks of "white" new tissue was detected in the defected site, HE histopathological analysis confirmed the new tissue was bone tissue). 28917901 1 59 theme cranial 229:235 arg1 regeneration 242:253 the complicated cranial bone regeneration 213:253 the complicated cranial bone regeneration 213:253 Bone disease is a public health problem around the word, and it is urgent to develop novel tissue engineering scaffolds for the complicated cranial bone regeneration. 27722232 6 0 theme general 1154:1160 arg1 utility 1162:1168 The general utility 1150:1168 The general utility of this approach 1150:1185 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 8 1 theme @ 1774:1774 arg1 DHB 1775:1777 HgTe@DHB 1770:1777 HgTe@DHB 1770:1777 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 3 2 theme @ 603:603 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 2 theme @ 603:603 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 8 3 theme relevant 1830:1837 arg1 carbohydrates 1860:1872 biologically relevant and more complicated carbohydrates 1817:1872 biologically relevant and more complicated carbohydrates 1817:1872 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 6 4 theme glycans 1294:1300 arg1 characterization 1211:1226 the characterization 1207:1226 the characterization of labile sialylated glycans 1207:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 4 theme glycans 1294:1300 arg1 glycans 1294:1300 complicated isomeric glycans 1273:1300 complicated isomeric glycans 1273:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 4 theme glycans 1294:1300 arg1 glycans 1249:1255 labile sialylated glycans 1231:1255 labile sialylated glycans 1231:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 4 theme glycans 1294:1300 arg1 sets 1265:1268 two sets 1261:1268 two sets of complicated isomeric glycans 1261:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 3 5 theme dual 618:621 arg1 element 647:653 a dual ionization-dissociation element 616:653 a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry 616:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 5 theme dual 618:621 arg1 acid 546:549 2,5-dihydroxybenzoic acid 525:549 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) 525:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 6 6 theme sialylated 1238:1247 arg1 glycans 1249:1255 labile sialylated glycans 1231:1255 labile sialylated glycans 1231:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 8 7 theme carbohydrates 1860:1872 arg1 analysis 1805:1812 the analysis 1801:1812 the analysis of biologically relevant and more complicated carbohydrates 1801:1872 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 4 8 theme laser-induced 765:777 arg1 dissociation 801:812 laser-induced extensive and intense dissociation 765:812 laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 765:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 3 9 theme matrix-assisted 658:672 arg1 spectrometry 707:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 7 10 theme release 1508:1514 arg1 dynamics 1516:1523 photoelectron release dynamics 1494:1523 photoelectron release dynamics 1494:1523 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 5 11 theme labile 1123:1128 arg1 glycans 1141:1147 labile sialylated glycans 1123:1147 labile sialylated glycans 1123:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 2 12 theme glycan 342:347 arg1 characterization 322:337 the structural characterization 307:337 the structural characterization of glycan 307:347 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 4 13 theme @ 748:748 arg1 NPs 753:755 HgTe@DHB NPs 744:755 HgTe@DHB NPs 744:755 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 5 14 theme glycans 1141:1147 arg1 linkage 1112:1118 linkage 1112:1118 linkage 1112:1118 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 14 theme glycans 1141:1147 arg1 sequence 1087:1094 sequence 1087:1094 sequence 1087:1094 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 14 theme glycans 1141:1147 arg1 composition 1074:1084 composition 1074:1084 composition 1074:1084 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 14 theme glycans 1141:1147 arg1 branching 1097:1105 branching 1097:1105 branching 1097:1105 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 2 15 theme high 270:273 arg1 degree 275:280 incredibly high degree 259:280 incredibly high degree of structural diversity 259:304 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 8 16 theme chemical 1896:1903 arg1 pre-derivatization 1905:1922 chemical pre-derivatization 1896:1922 chemical pre-derivatization 1896:1922 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 7 17 theme UV 1479:1480 arg1 absorption 1482:1491 UV absorption 1479:1491 UV absorption 1479:1491 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 6 18 gly sialylated 1238:1247 arg1 glycans 1249:1255 labile sialylated glycans 1231:1255 labile sialylated glycans 1231:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 3 19 theme desorption/ionization 680:700 arg1 spectrometry 707:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 6 20 theme complicated 1273:1283 arg1 glycans 1294:1300 complicated isomeric glycans 1273:1300 complicated isomeric glycans 1273:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 3 21 from element 647:653 arg1 spectrometry 707:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 5 22 theme linkage 1112:1118 arg1 determination 1053:1065 unambiguous determination 1041:1065 unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans 1041:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 2 23 theme diversity 296:304 arg1 degree 275:280 incredibly high degree 259:280 incredibly high degree of structural diversity 259:304 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 2 23 theme diversity 296:304 arg1 stability 245:253 low stability 241:253 low stability 241:253 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 8 24 theme tandem 1941:1946 arg1 spectrometry 1953:1964 conventional tandem mass spectrometry 1928:1964 conventional tandem mass spectrometry 1928:1964 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 5 25 theme cross-ring 994:1003 arg1 cleavage 1005:1012 cross-ring cleavage 994:1012 cross-ring cleavage (A-type ions) 994:1026 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 25 theme cross-ring 994:1003 arg1 ions 1022:1025 A-type ions 1015:1025 A-type ions 1015:1025 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 7 26 theme thermodynamic 1443:1455 arg1 absorption 1482:1491 UV absorption 1479:1491 UV absorption 1479:1491 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 26 theme thermodynamic 1443:1455 arg1 key 1554:1556 key 1554:1556 key 1554:1556 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 26 theme thermodynamic 1443:1455 arg1 properties 1457:1466 their nanoscale-dependent thermodynamic properties 1417:1466 their nanoscale-dependent thermodynamic properties 1417:1466 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 26 theme thermodynamic 1443:1455 arg1 energy 1537:1542 thermal energy 1529:1542 thermal energy 1529:1542 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 26 theme thermodynamic 1443:1455 arg1 dynamics 1516:1523 photoelectron release dynamics 1494:1523 photoelectron release dynamics 1494:1523 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 0 27 theme glycan 110:115 arg1 sequencing 117:126 comprehensive glycan sequencing 96:126 comprehensive glycan sequencing 96:126 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 1 28 theme various 198:204 arg1 processes 217:225 various biological processes 198:225 various biological processes 198:225 Glycoconjugates are ubiquitously present and play a critical role in various biological processes. 27722232 4 29 theme other 865:869 arg1 ZnO 902:904 ZnO 902:904 ZnO 902:904 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 29 theme other 865:869 arg1 TiO2 896:899 TiO2 896:899 TiO2 896:899 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 29 theme other 865:869 arg1 nanoparticles 881:893 other inorganic nanoparticles 865:893 other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 865:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 5 30 theme ions 1028:1031 arg1 generation 932:941 Abundant generation 923:941 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 923:1031 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 31 theme unambiguous 1041:1051 arg1 determination 1053:1065 unambiguous determination 1041:1065 unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans 1041:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 7 32 theme nanoscale-dependent 1423:1441 arg1 absorption 1482:1491 UV absorption 1479:1491 UV absorption 1479:1491 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 32 theme nanoscale-dependent 1423:1441 arg1 key 1554:1556 key 1554:1556 key 1554:1556 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 32 theme nanoscale-dependent 1423:1441 arg1 properties 1457:1466 their nanoscale-dependent thermodynamic properties 1417:1466 their nanoscale-dependent thermodynamic properties 1417:1466 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 32 theme nanoscale-dependent 1423:1441 arg1 energy 1537:1542 thermal energy 1529:1542 thermal energy 1529:1542 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 7 32 theme nanoscale-dependent 1423:1441 arg1 dynamics 1516:1523 photoelectron release dynamics 1494:1523 photoelectron release dynamics 1494:1523 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 0 33 theme Functionalized 0:13 arg1 nanoparticles 20:32 Functionalized HgTe nanoparticles 0:32 Functionalized HgTe nanoparticles 0:32 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 7 34 theme laser 1679:1683 arg1 process 1707:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 1 35 theme critical 181:188 arg1 role 190:193 a critical role 179:193 a critical role 179:193 Glycoconjugates are ubiquitously present and play a critical role in various biological processes. 27722232 5 36 dep glycosidic 957:966 arg1 Y- 969:970 Y- 969:970 Y- 969:970 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 36 dep glycosidic 957:966 arg1 ions 984:987 B-type ions 977:987 B-type ions 977:987 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 0 37 theme laser-induced 42:54 arg1 ionization/dissociation 68:90 laser-induced solid phase ionization/dissociation 42:90 laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing 42:126 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 5 38 theme sequence 1087:1094 arg1 determination 1053:1065 unambiguous determination 1041:1065 unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans 1041:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 7 39 theme physico-chemical 1368:1383 arg1 characteristics 1385:1399 the NP's physico-chemical characteristics 1359:1399 the NP's physico-chemical characteristics 1359:1399 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 3 40 theme complicated 458:468 arg1 characterization 477:492 complicated glycan characterization 458:492 complicated glycan characterization 458:492 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 0 41 theme phase 62:66 arg1 ionization/dissociation 68:90 laser-induced solid phase ionization/dissociation 42:90 laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing 42:126 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 5 42 theme branching 1097:1105 arg1 determination 1053:1065 unambiguous determination 1041:1065 unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans 1041:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 43 gly sialylated 1130:1139 arg1 glycans 1141:1147 labile sialylated glycans 1123:1147 labile sialylated glycans 1123:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 2 44 theme low 241:243 arg1 stability 245:253 low stability 241:253 low stability 241:253 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 3 45 theme mercury 566:572 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 45 theme mercury 566:572 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 7 46 theme spontaneous 1613:1623 arg1 decomposition 1632:1644 spontaneous glycan decomposition 1613:1644 spontaneous glycan decomposition 1613:1644 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 8 47 theme pseudo-MS/MS 1744:1755 arg1 " 1756:1756 this "pseudo-MS/MS" 1738:1756 this "pseudo-MS/MS" obtained by HgTe@DHB 1738:1777 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 3 48 theme DHB 604:606 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 48 theme DHB 604:606 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 5 49 theme composition 1074:1084 arg1 determination 1053:1065 unambiguous determination 1041:1065 unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans 1041:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 3 50 theme HgTe 599:602 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 50 theme HgTe 599:602 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 8 51 theme HgTe 1770:1773 arg1 DHB 1775:1777 HgTe@DHB 1770:1777 HgTe@DHB 1770:1777 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 6 52 theme glycans 1249:1255 arg1 characterization 1211:1226 the characterization 1207:1226 the characterization of labile sialylated glycans 1207:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 52 theme glycans 1249:1255 arg1 glycans 1294:1300 complicated isomeric glycans 1273:1300 complicated isomeric glycans 1273:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 52 theme glycans 1249:1255 arg1 glycans 1249:1255 labile sialylated glycans 1231:1255 labile sialylated glycans 1231:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 6 52 theme glycans 1249:1255 arg1 sets 1265:1268 two sets 1261:1268 two sets of complicated isomeric glycans 1261:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 2 53 theme structural 311:320 arg1 characterization 322:337 the structural characterization 307:337 the structural characterization of glycan 307:347 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 1 54 theme biological 206:215 arg1 processes 217:225 various biological processes 198:225 various biological processes 198:225 Glycoconjugates are ubiquitously present and play a critical role in various biological processes. 27722232 6 55 theme labile 1231:1236 arg1 glycans 1249:1255 labile sialylated glycans 1231:1255 labile sialylated glycans 1231:1255 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 4 56 theme DHB 749:751 arg1 NPs 753:755 HgTe@DHB NPs 744:755 HgTe@DHB NPs 744:755 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 2 57 theme chemical 368:375 arg1 derivatization 377:390 chemical derivatization 368:390 chemical derivatization 368:390 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 8 58 theme complicated 1848:1858 arg1 carbohydrates 1860:1872 biologically relevant and more complicated carbohydrates 1817:1872 biologically relevant and more complicated carbohydrates 1817:1872 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 5 59 theme glycosidic 957:966 arg1 ions 1028:1031 diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 946:1031 diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 946:1031 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 7 60 theme thermal 1529:1535 arg1 energy 1537:1542 thermal energy 1529:1542 thermal energy 1529:1542 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 3 61 theme ionization-dissociation 623:645 arg1 element 647:653 a dual ionization-dissociation element 616:653 a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry 616:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 61 theme ionization-dissociation 623:645 arg1 acid 546:549 2,5-dihydroxybenzoic acid 525:549 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) 525:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 5 62 theme B-type 977:982 arg1 ions 984:987 B-type ions 977:987 B-type ions 977:987 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 63 dep composition 1074:1084 arg1 the 1070:1072 the 1070:1072 the 1070:1072 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 64 theme sialylated 1130:1139 arg1 glycans 1141:1147 labile sialylated glycans 1123:1147 labile sialylated glycans 1123:1147 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 3 65 theme mass 702:705 arg1 spectrometry 707:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 5 66 theme Abundant 923:930 arg1 generation 932:941 Abundant generation 923:941 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 923:1031 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 7 67 theme photoelectron 1494:1506 arg1 dynamics 1516:1523 photoelectron release dynamics 1494:1523 photoelectron release dynamics 1494:1523 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 4 68 theme HgTe 744:747 arg1 NPs 753:755 HgTe@DHB NPs 744:755 HgTe@DHB NPs 744:755 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 3 69 theme laser 674:678 arg1 spectrometry 707:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 matrix-assisted laser desorption/ionization mass spectrometry 658:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 6 70 theme isomeric 1285:1292 arg1 glycans 1294:1300 complicated isomeric glycans 1273:1300 complicated isomeric glycans 1273:1300 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 4 71 theme glycan 821:826 arg1 dissociation 801:812 laser-induced extensive and intense dissociation 765:812 laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 765:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 0 72 theme comprehensive 96:108 arg1 sequencing 117:126 comprehensive glycan sequencing 96:126 comprehensive glycan sequencing 96:126 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 2 73 theme structural 285:294 arg1 diversity 296:304 structural diversity 285:304 structural diversity 285:304 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 8 74 theme conventional 1928:1939 arg1 spectrometry 1953:1964 conventional tandem mass spectrometry 1928:1964 conventional tandem mass spectrometry 1928:1964 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 4 75 dep nanoparticles 881:893 arg1 nanoparticles 881:893 other inorganic nanoparticles 865:893 other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 865:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 75 dep nanoparticles 881:893 arg1 ZnO 902:904 ZnO 902:904 ZnO 902:904 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 75 dep nanoparticles 881:893 arg1 TiO2 896:899 TiO2 896:899 TiO2 896:899 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 75 dep nanoparticles 881:893 arg1 NPs 917:919 Mn2O3 NPs 911:919 Mn2O3 NPs 911:919 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 76 theme extensive 779:787 arg1 dissociation 801:812 laser-induced extensive and intense dissociation 765:812 laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 765:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 8 77 theme mass 1948:1951 arg1 spectrometry 1953:1964 conventional tandem mass spectrometry 1928:1964 conventional tandem mass spectrometry 1928:1964 Our results show that this "pseudo-MS/MS" obtained by HgTe@DHB can be beneficial for the analysis of biologically relevant and more complicated carbohydrates, without the need for chemical pre-derivatization and conventional tandem mass spectrometry. 27722232 4 78 theme intense 793:799 arg1 dissociation 801:812 laser-induced extensive and intense dissociation 765:812 laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 765:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 79 theme inorganic 871:879 arg1 ZnO 902:904 ZnO 902:904 ZnO 902:904 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 79 theme inorganic 871:879 arg1 TiO2 896:899 TiO2 896:899 TiO2 896:899 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 4 79 theme inorganic 871:879 arg1 nanoparticles 881:893 other inorganic nanoparticles 865:893 other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs) 865:920 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 0 80 theme HgTe 15:18 arg1 nanoparticles 20:32 Functionalized HgTe nanoparticles 0:32 Functionalized HgTe nanoparticles 0:32 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 5 81 theme cleavage 1005:1012 arg1 ions 1028:1031 diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 946:1031 diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions 946:1031 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 4 82 theme superior 829:836 arg1 glycan 821:826 the glycan 817:826 the glycan 817:826 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 3 83 theme single 499:504 arg1 assay 506:510 a single assay 497:510 a single assay 497:510 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 5 84 theme A-type 1015:1020 arg1 cleavage 1005:1012 cross-ring cleavage 994:1012 cross-ring cleavage (A-type ions) 994:1026 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 5 84 theme A-type 1015:1020 arg1 ions 1022:1025 A-type ions 1015:1025 A-type ions 1015:1025 Abundant generation of diagnostic glycosidic (Y-, and B-type ions) and cross-ring cleavage (A-type ions) ions permits unambiguous determination of the composition, sequence, branching, and linkage of labile sialylated glycans. 27722232 4 85 theme HgTe 841:844 arg1 microparticles 846:859 HgTe microparticles 841:859 HgTe microparticles 841:859 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 3 86 theme functionalized 551:564 arg1 element 647:653 a dual ionization-dissociation element 616:653 a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry 616:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 86 theme functionalized 551:564 arg1 acid 546:549 2,5-dihydroxybenzoic acid 525:549 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) 525:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 0 87 theme solid 56:60 arg1 ionization/dissociation 68:90 laser-induced solid phase ionization/dissociation 42:90 laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing 42:126 Functionalized HgTe nanoparticles promote laser-induced solid phase ionization/dissociation for comprehensive glycan sequencing. 27722232 7 88 theme desorption-ionization 1685:1705 arg1 process 1707:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 3 89 theme 2,5-dihydroxybenzoic 525:544 arg1 element 647:653 a dual ionization-dissociation element 616:653 a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry 616:718 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 89 theme 2,5-dihydroxybenzoic 525:544 arg1 acid 546:549 2,5-dihydroxybenzoic acid 525:549 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) 525:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 7 90 theme nanoparticle-assisted 1657:1677 arg1 process 1707:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 the nanoparticle-assisted laser desorption-ionization process 1653:1713 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 3 91 theme glycan 470:475 arg1 characterization 477:492 complicated glycan characterization 458:492 complicated glycan characterization 458:492 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 4 92 theme Mn2O3 911:915 arg1 NPs 917:919 Mn2O3 NPs 911:919 Mn2O3 NPs 911:919 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 27722232 2 93 theme sophisticated 396:408 arg1 instrumentation 410:424 sophisticated instrumentation 396:424 sophisticated instrumentation 396:424 Due to their low stability and incredibly high degree of structural diversity, the structural characterization of glycan generally requires chemical derivatization and sophisticated instrumentation. 27722232 3 94 dep functionalized 551:564 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 94 dep functionalized 551:564 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 6 95 theme approach 1178:1185 arg1 utility 1162:1168 The general utility 1150:1168 The general utility of this approach 1150:1185 The general utility of this approach was demonstrated by the characterization of labile sialylated glycans and two sets of complicated isomeric glycans. 27722232 3 96 theme telluride 574:582 arg1 nanoparticles 584:596 mercury telluride nanoparticles 566:596 mercury telluride nanoparticles (HgTe@DHB NPs) 566:611 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 3 96 theme telluride 574:582 arg1 NPs 608:610 HgTe@DHB NPs 599:610 HgTe@DHB NPs 599:610 Herein, we report a method for complicated glycan characterization in a single assay by employing 2,5-dihydroxybenzoic acid functionalized mercury telluride nanoparticles (HgTe@DHB NPs) as a dual ionization-dissociation element in matrix-assisted laser desorption/ionization mass spectrometry. 27722232 7 97 theme glycan 1625:1630 arg1 decomposition 1632:1644 spontaneous glycan decomposition 1613:1644 spontaneous glycan decomposition 1613:1644 This phenomenon was delineated further by investigating the NP's physico-chemical characteristics, revealing that their nanoscale-dependent thermodynamic properties, including UV absorption, photoelectron release dynamics and thermal energy, were the key to levitate temperature synergistically, thus inducing spontaneous glycan decomposition during the nanoparticle-assisted laser desorption-ionization process. 27722232 4 98 theme linear 729:734 arg1 glycan 736:741 a linear glycan 727:741 a linear glycan 727:741 Using a linear glycan, HgTe@DHB NPs promote laser-induced extensive and intense dissociation of the glycan, superior to HgTe microparticles and other inorganic nanoparticles (TiO2, ZnO, and Mn2O3 NPs). 26270612 1 0 from interest 215:222 arg1 recognition 269:279 structurally defining lectin-carbohydrate recognition 227:279 structurally defining lectin-carbohydrate recognition 227:279 The physiological significance arising from translating information stored in glycans into cellular effects explains the interest in structurally defining lectin-carbohydrate recognition. 26270612 3 1 theme glycan 599:604 arg1 chains 606:611 galactose-terminated glycan chains 578:611 galactose-terminated glycan chains 578:611 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 5 2 theme deuteration 898:908 arg1 reduction 885:893 an effective reduction 872:893 an effective reduction of deuteration 872:908 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 2 3 theme divergence 427:436 arg1 origins 400:406 the origins 396:406 the origins of specificity and divergence 396:436 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 3 4 theme Cell 439:442 arg1 binding 444:450 Cell binding 439:450 Cell binding by using glycosylation mutants 439:481 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 6 5 theme protons 1024:1030 arg1 accessibility 1001:1013 solvent accessibility 993:1013 solvent accessibility of amide protons beyond this site 993:1047 In addition, evidence for changes in solvent accessibility of amide protons beyond this site was obtained. 26270612 4 6 theme key 691:693 arg1 contact 695:701 a key contact 689:701 a key contact for the carboxylic acid 689:725 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 4 6 theme key 691:693 arg1 Arg58 680:684 Arg58 680:684 Arg58 680:684 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 6 7 theme amide 1018:1022 arg1 protons 1024:1030 amide protons 1018:1030 amide protons 1018:1030 In addition, evidence for changes in solvent accessibility of amide protons beyond this site was obtained. 26270612 1 8 theme physiological 98:110 arg1 significance 112:123 The physiological significance 94:123 The physiological significance arising from translating information stored in glycans into cellular effects 94:200 The physiological significance arising from translating information stored in glycans into cellular effects explains the interest in structurally defining lectin-carbohydrate recognition. 26270612 2 9 from set 303:305 arg1 chicken 350:356 chicken 350:356 chicken 350:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 3 10 theme chicken 527:533 arg1 CG-8N 547:551 CG-8N 547:551 CG-8N 547:551 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 3 10 theme chicken 527:533 arg1 galectin-8 535:544 chicken galectin-8 527:544 chicken galectin-8 (CG-8N) 527:552 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 3 11 theme glycosylation 461:473 arg1 mutants 475:481 glycosylation mutants 461:481 glycosylation mutants 461:481 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 3 12 theme N-terminal 506:515 arg1 domain 517:522 the N-terminal domain 502:522 the N-terminal domain of chicken galectin-8 (CG-8N) 502:552 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 3 12 theme N-terminal 506:515 arg1 galectin-8 535:544 chicken galectin-8 527:544 chicken galectin-8 (CG-8N) 527:552 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 5 13 theme contact 942:948 arg1 area 950:953 the contact area 938:953 the contact area 938:953 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 6 14 from changes 982:988 arg1 accessibility 1001:1013 solvent accessibility 993:1013 solvent accessibility of amide protons beyond this site 993:1047 In addition, evidence for changes in solvent accessibility of amide protons beyond this site was obtained. 26270612 5 15 theme mass 845:848 arg1 spectrometry 850:861 mass spectrometry 845:861 mass spectrometry 845:861 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 3 16 theme domain 517:522 arg1 binding 491:497 binding 491:497 binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains 491:611 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 0 17 theme Hydrogen-Deuterium 30:47 arg1 Exchange 49:56 Hydrogen-Deuterium Exchange 30:56 Hydrogen-Deuterium Exchange 30:56 Combining Crystallography and Hydrogen-Deuterium Exchange to Study Galectin-Ligand Complexes. 26270612 4 18 theme 3'-sialylated 646:658 arg1 derivative 660:669 its 3'-sialylated derivative 642:669 its 3'-sialylated derivative 642:669 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 5 19 theme Monitoring 803:812 arg1 exchange 833:840 Monitoring hydrogen-deuterium exchange 803:840 Monitoring hydrogen-deuterium exchange by mass spectrometry 803:861 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 1 20 theme defining 240:247 arg1 recognition 269:279 structurally defining lectin-carbohydrate recognition 227:279 structurally defining lectin-carbohydrate recognition 227:279 The physiological significance arising from translating information stored in glycans into cellular effects explains the interest in structurally defining lectin-carbohydrate recognition. 26270612 5 21 theme hydrogen-deuterium 814:831 arg1 exchange 833:840 Monitoring hydrogen-deuterium exchange 803:840 Monitoring hydrogen-deuterium exchange by mass spectrometry 803:861 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 6 22 theme solvent 993:999 arg1 accessibility 1001:1013 solvent accessibility 993:1013 solvent accessibility of amide protons beyond this site 993:1047 In addition, evidence for changes in solvent accessibility of amide protons beyond this site was obtained. 26270612 4 23 from differences 731:741 arg1 lengths 751:757 loop lengths 746:757 loop lengths to the three homodimeric chicken galectins 746:800 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 1 24 theme lectin-carbohydrate 249:267 arg1 recognition 269:279 structurally defining lectin-carbohydrate recognition 227:279 structurally defining lectin-carbohydrate recognition 227:279 The physiological significance arising from translating information stored in glycans into cellular effects explains the interest in structurally defining lectin-carbohydrate recognition. 26270612 3 25 theme galactose-terminated 578:597 arg1 chains 606:611 galactose-terminated glycan chains 578:611 galactose-terminated glycan chains 578:611 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 4 26 theme loop 746:749 arg1 lengths 751:757 loop lengths 746:757 loop lengths to the three homodimeric chicken galectins 746:800 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 2 27 from galectins 337:345 arg1 chicken 350:356 chicken 350:356 chicken 350:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 3 28 theme galectin-8 535:544 arg1 domain 517:522 the N-terminal domain 502:522 the N-terminal domain of chicken galectin-8 (CG-8N) 502:552 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 3 28 theme galectin-8 535:544 arg1 galectin-8 535:544 chicken galectin-8 527:544 chicken galectin-8 (CG-8N) 527:552 Cell binding by using glycosylation mutants reveals binding of the N-terminal domain of chicken galectin-8 (CG-8N) to α-2,3-sialylated and galactose-terminated glycan chains. 26270612 5 29 theme ligand 916:921 arg1 binding 923:929 ligand binding 916:929 ligand binding within the contact area 916:953 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 2 30 theme small 297:301 arg1 set 303:305 The relatively small set 282:305 The relatively small set of adhesion/growth-regulatory galectins in chicken 282:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 2 30 theme small 297:301 arg1 galectins 337:345 adhesion/growth-regulatory galectins 310:345 adhesion/growth-regulatory galectins in chicken 310:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 4 31 gly 3'-sialylated 646:658 arg1 derivative 660:669 its 3'-sialylated derivative 642:669 its 3'-sialylated derivative 642:669 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 0 32 theme Galectin-Ligand 67:81 arg1 Complexes 83:91 Study Galectin-Ligand Complexes 61:91 Study Galectin-Ligand Complexes 61:91 Combining Crystallography and Hydrogen-Deuterium Exchange to Study Galectin-Ligand Complexes. 26270612 4 33 from contact 695:701 arg1 lengths 751:757 loop lengths 746:757 loop lengths to the three homodimeric chicken galectins 746:800 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 7 34 theme systematic 1147:1156 arg1 studies 1158:1164 systematic studies 1147:1164 systematic studies on galectins and beyond 1147:1188 Their detection, which highlights the sensor capacity of this technique, encourages systematic studies on galectins and beyond. 26270612 4 35 with Cocrystals 614:623 arg1 derivative 660:669 its 3'-sialylated derivative 642:669 its 3'-sialylated derivative 642:669 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 4 35 with Cocrystals 614:623 arg1 lactose 630:636 lactose 630:636 lactose 630:636 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 0 36 theme Study 61:65 arg1 Complexes 83:91 Study Galectin-Ligand Complexes 61:91 Study Galectin-Ligand Complexes 61:91 Combining Crystallography and Hydrogen-Deuterium Exchange to Study Galectin-Ligand Complexes. 26270612 7 37 theme technique 1125:1133 arg1 capacity 1108:1115 the sensor capacity 1097:1115 the sensor capacity of this technique 1097:1133 Their detection, which highlights the sensor capacity of this technique, encourages systematic studies on galectins and beyond. 26270612 2 38 theme specificity 411:421 arg1 origins 400:406 the origins 396:406 the origins of specificity and divergence 396:436 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 2 39 from chicken 350:356 arg1 set 303:305 The relatively small set 282:305 The relatively small set of adhesion/growth-regulatory galectins in chicken 282:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 2 39 from chicken 350:356 arg1 galectins 337:345 adhesion/growth-regulatory galectins 310:345 adhesion/growth-regulatory galectins in chicken 310:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 7 40 from studies 1158:1164 arg1 galectins 1169:1177 galectins 1169:1177 galectins 1169:1177 Their detection, which highlights the sensor capacity of this technique, encourages systematic studies on galectins and beyond. 26270612 7 40 from studies 1158:1164 arg1 beyond 1183:1188 beyond 1183:1188 beyond 1183:1188 Their detection, which highlights the sensor capacity of this technique, encourages systematic studies on galectins and beyond. 26270612 2 41 theme galectins 337:345 arg1 set 303:305 The relatively small set 282:305 The relatively small set of adhesion/growth-regulatory galectins in chicken 282:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 2 41 theme galectins 337:345 arg1 galectins 337:345 adhesion/growth-regulatory galectins 310:345 adhesion/growth-regulatory galectins in chicken 310:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 4 42 theme homodimeric 772:782 arg1 galectins 792:800 the three homodimeric chicken galectins 762:800 the three homodimeric chicken galectins 762:800 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 5 43 theme effective 875:883 arg1 reduction 885:893 an effective reduction 872:893 an effective reduction of deuteration 872:908 Monitoring hydrogen-deuterium exchange by mass spectrometry revealed an effective reduction of deuteration after ligand binding within the contact area. 26270612 2 44 theme adhesion/growth-regulatory 310:335 arg1 galectins 337:345 adhesion/growth-regulatory galectins 310:345 adhesion/growth-regulatory galectins in chicken 310:356 The relatively small set of adhesion/growth-regulatory galectins in chicken makes this system attractive to study the origins of specificity and divergence. 26270612 4 45 theme chicken 784:790 arg1 galectins 792:800 the three homodimeric chicken galectins 762:800 the three homodimeric chicken galectins 762:800 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 26270612 7 46 theme sensor 1101:1106 arg1 capacity 1108:1115 the sensor capacity 1097:1115 the sensor capacity of this technique 1097:1133 Their detection, which highlights the sensor capacity of this technique, encourages systematic studies on galectins and beyond. 26270612 1 47 theme cellular 185:192 arg1 effects 194:200 cellular effects 185:200 cellular effects 185:200 The physiological significance arising from translating information stored in glycans into cellular effects explains the interest in structurally defining lectin-carbohydrate recognition. 26270612 4 48 theme carboxylic 711:720 arg1 acid 722:725 the carboxylic acid 707:725 the carboxylic acid 707:725 Cocrystals with lactose and its 3'-sialylated derivative disclose Arg58 as a key contact for the carboxylic acid and differences in loop lengths to the three homodimeric chicken galectins. 25175058 11 0 contain has 1953:1955 arg1 Identification 1862:1875 Identification 1862:1875 Identification of individual monomeric units, linkages, ring size, branching and anomerity 1862:1951 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 0 contain has 1953:1955 arg2 challenges 1975:1984 posed significant challenges 1957:1984 posed significant challenges 1957:1984 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 6 1 theme complexes 876:884 arg1 stability 823:831 relative stability 814:831 relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase 814:901 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 14 2 theme structural 2362:2371 arg1 details 2373:2379 structural details 2362:2379 structural details 2362:2379 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 6 3 from stability 823:831 arg1 phase 897:901 the gas phase 889:901 the gas phase 889:901 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 10 4 theme modification 1669:1680 arg1 aspects 1631:1637 manifold aspects 1622:1637 manifold aspects of post-translational protein modification 1622:1680 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 6 5 theme glycosidic 836:845 arg1 bonds 847:851 glycosidic bonds 836:851 glycosidic bonds 836:851 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 9 6 from lines 1422:1426 arg1 activities 1367:1376 expressed activities 1357:1376 expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) 1357:1466 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 12 7 theme growing 2022:2028 arg1 method 2047:2052 a growing key instrumental method 2020:2052 a growing key instrumental method to differentiate among isomers [12] 2020:2088 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 12 7 theme growing 2022:2028 arg1 MS 2011:2012 MS 2011:2012 MS(n) 2011:2015 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 19 8 theme potential 3280:3288 arg1 structures 3290:3299 potential structures 3280:3299 potential structures 3280:3299 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 19 9 theme altered 3370:3376 arg1 structures 3378:3387 altered structures 3370:3387 altered structures 3370:3387 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 2 10 theme successive 231:240 arg1 fragmentations 242:255 successive fragmentations 231:255 successive fragmentations of sodiated oligosaccharides 231:284 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 15 11 theme fundamental 2493:2503 arg1 need 2505:2508 a fundamental need 2491:2508 a fundamental need for high throughput tools in glycomics to complement proteome studies 2491:2578 There is a fundamental need for high throughput tools in glycomics to complement proteome studies. 25175058 8 12 theme linkage 1165:1171 arg1 determination 1173:1185 linkage determination 1165:1185 linkage determination of carbohydrates aided by a "B2 library" 1165:1226 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 18 13 theme energy 2960:2965 arg1 independence 2967:2978 energy independence 2960:2978 energy independence from modes of ionization and collisional activation 2960:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 9 14 from transferases 1394:1405 arg1 lines 1422:1426 cancer cell lines 1410:1426 cancer cell lines 1410:1426 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 2 15 theme oligosaccharides 269:284 arg1 fragmentations 242:255 successive fragmentations 231:255 successive fragmentations of sodiated oligosaccharides 231:284 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 9 16 theme activities 1367:1376 arg1 levels 1347:1352 levels 1347:1352 levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ 1347:1516 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 11 17 theme posed 1957:1961 arg1 challenges 1975:1984 posed significant challenges 1957:1984 posed significant challenges 1957:1984 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 15 18 from tools 2530:2534 arg1 glycomics 2539:2547 glycomics 2539:2547 glycomics 2539:2547 There is a fundamental need for high throughput tools in glycomics to complement proteome studies. 25175058 10 19 theme BIOLOGICAL 1519:1528 arg1 SIGNIFICANCE 1530:1541 BIOLOGICAL SIGNIFICANCE 1519:1541 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 7 20 theme B2 1002:1003 arg1 information 1009:1019 B2 ion information 1002:1019 B2 ion information 1002:1019 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 11 21 theme individual 1880:1889 arg1 size 1923:1926 ring size 1918:1926 ring size 1918:1926 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 21 theme individual 1880:1889 arg1 units 1901:1905 individual monomeric units 1880:1905 individual monomeric units 1880:1905 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 21 theme individual 1880:1889 arg1 anomerity 1943:1951 anomerity 1943:1951 anomerity 1943:1951 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 21 theme individual 1880:1889 arg1 branching 1929:1937 branching 1929:1937 branching 1929:1937 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 21 theme individual 1880:1889 arg1 linkages 1908:1915 linkages 1908:1915 linkages 1908:1915 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 16 22 from regard 2589:2594 arg1 important 2619:2627 important 2619:2627 important 2619:2627 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 12 23 dep isomers 2077:2083 arg1 [12 2085:2087 [12 2085:2087 [12 2085:2087 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 7 24 from scarcity 1124:1131 arg1 nature 1136:1141 nature 1136:1141 nature 1136:1141 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 16 25 theme searchable 2634:2643 arg1 files 2662:2666 searchable spectral library files 2634:2666 searchable spectral library files for structural confirmation 2634:2694 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 17 26 theme carbohydrates 2829:2841 arg1 structures 2815:2824 more than 10,000 synthetic structures 2788:2824 more than 10,000 synthetic structures of carbohydrates 2788:2841 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 9 27 theme transferases 1394:1405 arg1 activities 1367:1376 expressed activities 1357:1376 expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) 1357:1466 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 11 28 theme mass 1989:1992 arg1 spectrometrists 1994:2008 mass spectrometrists 1989:2008 mass spectrometrists 1989:2008 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 16 29 theme library 2654:2660 arg1 files 2662:2666 searchable spectral library files 2634:2666 searchable spectral library files for structural confirmation 2634:2694 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 4 30 theme B2 672:673 arg1 " 682:682 a "B2 library" 669:682 a "B2 library" 669:682 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 19 31 theme B2 3158:3159 arg1 library 3161:3167 the new B2 library 3150:3167 the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases 3150:3256 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 7 32 theme linear 921:926 arg1 trisaccharides 938:951 linear, branched trisaccharides 921:951 linear, branched trisaccharides 921:951 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 9 33 theme cancer 1410:1415 arg1 lines 1422:1426 cancer cell lines 1410:1426 cancer cell lines 1410:1426 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 8 34 theme B2 1216:1217 arg1 " 1226:1226 a "B2 library" 1213:1226 a "B2 library" 1213:1226 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 5 35 theme characteristic 702:715 arg1 patterns 731:738 characteristic fragmentation patterns 702:738 characteristic fragmentation patterns 702:738 Linkages produce characteristic fragmentation patterns in the absence of cross-ring fragmentation. 25175058 2 36 theme ion-trap 203:210 arg1 spectrometer 217:228 a quadrupole ion-trap mass spectrometer 190:228 a quadrupole ion-trap mass spectrometer 190:228 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 17 37 theme synthetic 2805:2813 arg1 structures 2815:2824 more than 10,000 synthetic structures 2788:2824 more than 10,000 synthetic structures of carbohydrates 2788:2841 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 11 38 theme monomeric 1891:1899 arg1 size 1923:1926 ring size 1918:1926 ring size 1918:1926 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 38 theme monomeric 1891:1899 arg1 units 1901:1905 individual monomeric units 1880:1905 individual monomeric units 1880:1905 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 38 theme monomeric 1891:1899 arg1 anomerity 1943:1951 anomerity 1943:1951 anomerity 1943:1951 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 38 theme monomeric 1891:1899 arg1 branching 1929:1937 branching 1929:1937 branching 1929:1937 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 38 theme monomeric 1891:1899 arg1 linkages 1908:1915 linkages 1908:1915 linkages 1908:1915 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 18 39 theme trap 2942:2945 arg1 spectra 2947:2953 ion trap spectra 2938:2953 ion trap spectra 2938:2953 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 14 40 theme ID 2430:2431 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 17 41 theme National 2701:2708 arg1 Academy 2710:2716 The National Academy 2697:2716 The National Academy of Science (NAS) report (http://glyco.nas.edu) 2697:2763 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 7 42 from absence 1080:1086 arg1 sequence 1103:1110 an isomeric sequence 1091:1110 an isomeric sequence 1091:1110 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 17 43 theme glycomics 2867:2875 arg1 field 2858:2862 the field 2854:2862 the field of glycomics 2854:2875 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 7 44 theme MS 904:905 arg1 studies 910:916 MS(n) studies 904:916 MS(n) studies of linear, branched trisaccharides and tetrasaccharides 904:972 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 1 45 theme mass 108:111 arg1 libraries 122:130 Searchable mass spectral libraries 97:130 Searchable mass spectral libraries for glycans 97:142 Searchable mass spectral libraries for glycans may be enhanced using a B2 ion library. 25175058 10 46 theme important 1563:1571 arg1 Glycosylation 1543:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 46 theme important 1563:1571 arg1 class 1573:1577 an important class 1560:1577 an important class 1560:1577 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 0 47 theme ion 75:77 arg1 library 88:94 A b2 ion spectral library 70:94 A b2 ion spectral library 70:94 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 17 48 theme structures 2815:2824 arg1 need 2780:2783 the need 2776:2783 the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics 2776:2875 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 15 49 theme proteome 2563:2570 arg1 studies 2572:2578 proteome studies 2563:2578 proteome studies 2563:2578 There is a fundamental need for high throughput tools in glycomics to complement proteome studies. 25175058 5 50 theme fragmentation 769:781 arg1 absence 747:753 the absence 743:753 the absence of cross-ring fragmentation 743:781 Linkages produce characteristic fragmentation patterns in the absence of cross-ring fragmentation. 25175058 18 51 theme achievable 3100:3109 arg1 [26 3127:3129 an achievable research target [26 3097:3129 an achievable research target [26 3097:3129 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 4 52 theme glycosidic 513:522 arg1 linkages 524:531 glycosidic linkages 513:531 glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, 513:599 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 4 52 theme glycosidic 513:522 arg1 Hex-Fuc 541:547 Hex-Fuc 541:547 Hex-Fuc 541:547 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 6 53 theme relative 814:821 arg1 stability 823:831 relative stability 814:831 relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase 814:901 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 17 54 dep http 2743:2746 arg1 //glyco.nas.edu 2748:2762 //glyco.nas.edu 2748:2762 http://glyco.nas.edu 2743:2762 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 1 55 theme spectral 113:120 arg1 libraries 122:130 Searchable mass spectral libraries 97:130 Searchable mass spectral libraries for glycans 97:142 Searchable mass spectral libraries for glycans may be enhanced using a B2 ion library. 25175058 15 56 theme high 2514:2517 arg1 tools 2530:2534 high throughput tools 2514:2534 high throughput tools in glycomics 2514:2547 There is a fundamental need for high throughput tools in glycomics to complement proteome studies. 25175058 10 57 theme unique 1775:1780 arg1 [6,7 1791:1794 unique haptenic [6,7] 1775:1795 unique haptenic [6,7] 1775:1795 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 18 58 theme general 2911:2917 arg1 reproducibility 2919:2933 the general reproducibility 2907:2933 the general reproducibility of ion trap spectra 2907:2953 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 3 59 theme specific 411:418 arg1 linkages 431:438 specific glycosidic linkages 411:438 specific glycosidic linkages using progressive MS stages 411:466 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 3 60 theme disaccharide 356:367 arg1 ions 372:375 disaccharide B2 ions 356:375 disaccharide B2 ions 356:375 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 4 61 theme ions 490:493 arg1 " 494:494 "B2 ions" 486:494 "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, 486:599 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 2 62 theme ion 319:321 arg1 mode 323:326 the positive ion mode 306:326 the positive ion mode 306:326 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 0 63 theme spectrometry 21:32 arg1 strategy 34:41 MS(n) mass spectrometry strategy 10:41 MS(n) mass spectrometry strategy for carbohydrate analysis 10:67 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 13 64 theme potential 2101:2109 arg1 isomers 2111:2117 the potential isomers 2097:2117 the potential isomers in oligosaccharides 2097:2137 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 6 65 theme ions 800:803 arg1 " 804:804 "B2 ions" 796:804 "B2 ions" 796:804 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 18 66 theme activation 3021:3030 arg1 modes 2985:2989 modes 2985:2989 modes of ionization and collisional activation 2985:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 11 67 theme units 1901:1905 arg1 Identification 1862:1875 Identification 1862:1875 Identification of individual monomeric units, linkages, ring size, branching and anomerity 1862:1951 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 14 68 theme collision 2322:2330 arg1 activation 2332:2341 collision activation 2322:2341 collision activation (MS(n)) 2322:2349 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 14 68 theme collision 2322:2330 arg1 MS 2344:2345 MS 2344:2345 MS(n) 2344:2348 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 10 69 theme " 1604:1604 arg1 Glycosylation 1543:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 69 theme " 1604:1604 arg1 class 1573:1577 an important class 1560:1577 an important class 1560:1577 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 70 theme post-translational 1642:1659 arg1 modification 1669:1680 post-translational protein modification 1642:1680 post-translational protein modification 1642:1680 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 14 71 theme precise 2385:2391 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 13 72 theme glycosyl 2252:2259 arg1 transferases 2261:2272 the expressed glycosyl transferases [13-20] 2238:2280 the expressed glycosyl transferases [13-20] 2238:2280 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 16 73 from important 2619:2627 arg1 regard 2589:2594 that regard 2584:2594 that regard 2584:2594 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 14 74 theme sequential 2301:2310 arg1 stages 2312:2317 sequential stages 2301:2317 sequential stages of collision activation (MS(n)) 2301:2349 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 19 75 theme structures 3378:3387 arg1 cohort 3360:3365 the expected cohort 3347:3365 the expected cohort of altered structures 3347:3387 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 0 76 dep Enhancing 0:8 arg1 library 88:94 A b2 ion spectral library 70:94 A b2 ion spectral library 70:94 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 18 77 theme ion 2938:2940 arg1 spectra 2947:2953 ion trap spectra 2938:2953 ion trap spectra 2938:2953 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 10 78 theme oncology 1822:1829 arg1 [8-11 1831:1835 oncology [8-11] 1822:1836 oncology [8-11] 1822:1836 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 79 theme manifold 1622:1629 arg1 aspects 1631:1637 manifold aspects 1622:1637 manifold aspects of post-translational protein modification 1622:1680 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 6 80 theme carbohydrate-metal 857:874 arg1 complexes 876:884 carbohydrate-metal complexes 857:884 carbohydrate-metal complexes 857:884 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 14 81 theme linkage 2413:2419 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 14 82 theme Mass 2283:2286 arg1 spectra 2288:2294 Mass spectra 2283:2294 Mass spectra from sequential stages of collision activation (MS(n)) 2283:2349 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 2 83 theme sodiated 260:267 arg1 oligosaccharides 269:284 sodiated oligosaccharides 260:284 sodiated oligosaccharides 260:284 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 6 84 theme bonds 847:851 arg1 stability 823:831 relative stability 814:831 relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase 814:901 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 16 85 theme structural 2672:2681 arg1 confirmation 2683:2694 structural confirmation 2672:2694 structural confirmation 2672:2694 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 10 86 theme other 1842:1846 arg1 applications 1848:1859 other applications 1842:1859 other applications 1842:1859 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 5 87 theme cross-ring 758:767 arg1 fragmentation 769:781 cross-ring fragmentation 758:781 cross-ring fragmentation 758:781 Linkages produce characteristic fragmentation patterns in the absence of cross-ring fragmentation. 25175058 10 88 theme protein 1661:1667 arg1 modification 1669:1680 post-translational protein modification 1642:1680 post-translational protein modification 1642:1680 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 4 89 theme energy 636:641 arg1 CID 643:645 low energy CID 632:645 low energy CID 632:645 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 12 90 theme instrumental 2034:2045 arg1 method 2047:2052 a growing key instrumental method 2020:2052 a growing key instrumental method to differentiate among isomers [12] 2020:2088 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 12 90 theme instrumental 2034:2045 arg1 MS 2011:2012 MS 2011:2012 MS(n) 2011:2015 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 8 91 theme MS 1149:1150 arg1 strategy 1152:1159 This MS strategy 1144:1159 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" 1144:1226 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 19 92 theme structures 3290:3299 arg1 classes 3269:3275 classes 3269:3275 classes of potential structures 3269:3299 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 18 93 theme target 3120:3125 arg1 [26 3127:3129 an achievable research target [26 3097:3129 an achievable research target [26 3097:3129 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 4 94 theme low 632:634 arg1 CID 643:645 low energy CID 632:645 low energy CID 632:645 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 7 95 theme ion 1005:1007 arg1 information 1009:1019 B2 ion information 1002:1019 B2 ion information 1002:1019 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 10 96 theme protein 1693:1699 arg1 conformation 1701:1712 protein conformation 1693:1712 protein conformation 1693:1712 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 9 97 theme expressed 1357:1365 arg1 activities 1367:1376 expressed activities 1357:1376 expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) 1357:1466 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 18 98 from modes 2985:2989 arg1 reproducibility 2919:2933 the general reproducibility 2907:2933 the general reproducibility of ion trap spectra 2907:2953 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 18 98 from modes 2985:2989 arg1 independence 2967:2978 energy independence 2960:2978 energy independence from modes of ionization and collisional activation 2960:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 19 99 theme cohort 3360:3365 arg1 identification 3329:3342 MS identification 3326:3342 MS identification of the expected cohort of altered structures 3326:3387 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 10 100 theme SIGNIFICANCE 1530:1541 arg1 Glycosylation 1543:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation 1519:1555 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 100 theme SIGNIFICANCE 1530:1541 arg1 class 1573:1577 an important class 1560:1577 an important class 1560:1577 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 101 theme protein 1737:1743 arg1 receptors 1745:1753 protein receptors [1-5] 1737:1759 protein receptors [1-5] 1737:1759 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 12 102 theme key 2030:2032 arg1 method 2047:2052 a growing key instrumental method 2020:2052 a growing key instrumental method to differentiate among isomers [12] 2020:2088 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 12 102 theme key 2030:2032 arg1 MS 2011:2012 MS 2011:2012 MS(n) 2011:2015 MS(n) is a growing key instrumental method to differentiate among isomers [12]. 25175058 1 103 theme ion 171:173 arg1 library 175:181 a B2 ion library 166:181 a B2 ion library 166:181 Searchable mass spectral libraries for glycans may be enhanced using a B2 ion library. 25175058 11 104 theme significant 1963:1973 arg1 challenges 1975:1984 posed significant challenges 1957:1984 posed significant challenges 1957:1984 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 8 105 theme carbohydrates 1190:1202 arg1 determination 1173:1185 linkage determination 1165:1185 linkage determination of carbohydrates aided by a "B2 library" 1165:1226 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 19 106 theme expected 3351:3358 arg1 cohort 3360:3365 the expected cohort 3347:3365 the expected cohort of altered structures 3347:3387 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 7 107 theme tetrasaccharides 957:972 arg1 studies 910:916 MS(n) studies 904:916 MS(n) studies of linear, branched trisaccharides and tetrasaccharides 904:972 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 9 108 theme glycosyl 1385:1392 arg1 transferases 1394:1405 two glycosyl transferases 1381:1405 two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) 1381:1466 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 6 109 theme gas 893:895 arg1 phase 897:901 the gas phase 889:901 the gas phase 889:901 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 19 110 theme carbohydrate 3173:3184 arg1 differences 3186:3196 carbohydrate differences 3173:3196 carbohydrate differences found on cancers, where we profile the glycosyltransferases 3173:3256 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 19 111 located found 3198:3202 arg2 differences 3186:3196 carbohydrate differences 3173:3196 carbohydrate differences found on cancers, where we profile the glycosyltransferases 3173:3256 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 19 111 located found 3198:3202 arg1 cancers 3207:3213 cancers 3207:3213 cancers 3207:3213 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 7 112 theme trisaccharides 938:951 arg1 studies 910:916 MS(n) studies 904:916 MS(n) studies of linear, branched trisaccharides and tetrasaccharides 904:972 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 1 113 theme Searchable 97:106 arg1 libraries 122:130 Searchable mass spectral libraries 97:130 Searchable mass spectral libraries for glycans 97:142 Searchable mass spectral libraries for glycans may be enhanced using a B2 ion library. 25175058 19 114 theme MS 3326:3327 arg1 identification 3329:3342 MS identification 3326:3342 MS identification of the expected cohort of altered structures 3326:3387 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 16 115 theme spectral 2645:2652 arg1 files 2662:2666 searchable spectral library files 2634:2666 searchable spectral library files for structural confirmation 2634:2694 In that regard, nothing could be more important than searchable spectral library files for structural confirmation. 25175058 3 116 theme progressive 446:456 arg1 stages 461:466 progressive MS stages 446:466 progressive MS stages 446:466 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 0 117 theme carbohydrate 47:58 arg1 analysis 60:67 carbohydrate analysis 47:67 carbohydrate analysis 47:67 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 2 118 theme mass 212:215 arg1 spectrometer 217:228 a quadrupole ion-trap mass spectrometer 190:228 a quadrupole ion-trap mass spectrometer 190:228 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 18 119 theme collisional 3009:3019 arg1 activation 3021:3030 collisional activation 3009:3030 collisional activation 3009:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 9 120 theme cell 1417:1420 arg1 lines 1422:1426 cancer cell lines 1410:1426 cancer cell lines 1410:1426 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 14 121 theme branching 2463:2471 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 14 122 from stages 2312:2317 arg1 spectra 2288:2294 Mass spectra 2283:2294 Mass spectra from sequential stages of collision activation (MS(n)) 2283:2349 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 8 123 theme library 1219:1225 arg1 " 1226:1226 a "B2 library" 1213:1226 a "B2 library" 1213:1226 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 19 124 theme new 3154:3156 arg1 library 3161:3167 the new B2 library 3150:3167 the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases 3150:3256 We intend to use the new B2 library for carbohydrate differences found on cancers, where we profile the glycosyltransferases to predict classes of potential structures, and use the library for MS identification of the expected cohort of altered structures. 25175058 5 125 theme fragmentation 717:729 arg1 patterns 731:738 characteristic fragmentation patterns 702:738 characteristic fragmentation patterns 702:738 Linkages produce characteristic fragmentation patterns in the absence of cross-ring fragmentation. 25175058 18 126 theme spectra 2947:2953 arg1 reproducibility 2919:2933 the general reproducibility 2907:2933 the general reproducibility of ion trap spectra 2907:2953 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 18 126 theme spectra 2947:2953 arg1 independence 2967:2978 energy independence 2960:2978 energy independence from modes of ionization and collisional activation 2960:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 14 127 theme anomerity 2449:2457 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 4 128 theme B2 487:488 arg1 ions 490:493 B2 ions 487:493 "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, 486:599 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 13 129 from isomers 2111:2117 arg1 oligosaccharides 2122:2137 oligosaccharides 2122:2137 oligosaccharides 2122:2137 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 0 130 theme b2 72:73 arg1 library 88:94 A b2 ion spectral library 70:94 A b2 ion spectral library 70:94 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 18 131 theme carbohydrate 3069:3080 arg1 identification 3082:3095 carbohydrate identification 3069:3095 carbohydrate identification 3069:3095 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 14 132 theme substitutions 2434:2446 arg1 characterization 2393:2408 precise characterization 2385:2408 precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25] 2385:2479 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 10 133 dep includes 1613:1620 arg1 providing 1715:1723 providing 1715:1723 providing ligands for protein receptors [1-5] 1715:1759 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 133 dep includes 1613:1620 arg1 encoding 1766:1773 encoding 1766:1773 encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications 1766:1859 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 133 dep includes 1613:1620 arg1 affecting 1683:1691 affecting 1683:1691 affecting protein conformation 1683:1712 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 10 134 theme postranslationome 1587:1603 arg1 " 1604:1604 the "postranslationome" 1582:1604 the "postranslationome" 1582:1604 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 0 135 theme spectral 79:86 arg1 library 88:94 A b2 ion spectral library 70:94 A b2 ion spectral library 70:94 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 1 136 theme B2 168:169 arg1 library 175:181 a B2 ion library 166:181 a B2 ion library 166:181 Searchable mass spectral libraries for glycans may be enhanced using a B2 ion library. 25175058 17 137 theme Science 2721:2727 arg1 Academy 2710:2716 The National Academy 2697:2716 The National Academy of Science (NAS) report (http://glyco.nas.edu) 2697:2763 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 9 138 dep transferases 1394:1405 arg1 B4GALNT3&4 1456:1465 B4GALNT3&4 1456:1465 B4GALNT3&4 1456:1465 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 9 138 dep transferases 1394:1405 arg1 B3GALNT2 1432:1439 B3GALNT2 1432:1439 B3GALNT2 1432:1439 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 9 138 dep transferases 1394:1405 arg1 β4GalNAcT 1446:1454 β4GalNAcT 1446:1454 β4GalNAcT(B4GALNT3&4) 1446:1466 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 9 138 dep transferases 1394:1405 arg1 β3 1429:1430 β3 1429:1430 β3(B3GALNT2) 1429:1440 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 15 139 theme throughput 2519:2528 arg1 tools 2530:2534 high throughput tools 2514:2534 high throughput tools in glycomics 2514:2547 There is a fundamental need for high throughput tools in glycomics to complement proteome studies. 25175058 0 140 theme MS 10:11 arg1 strategy 34:41 MS(n) mass spectrometry strategy 10:41 MS(n) mass spectrometry strategy for carbohydrate analysis 10:67 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 18 141 theme research 3111:3118 arg1 [26 3127:3129 an achievable research target [26 3097:3129 an achievable research target [26 3097:3129 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 4 142 theme library 675:681 arg1 " 682:682 a "B2 library" 669:682 a "B2 library" 669:682 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 0 143 theme n 13:13 arg1 strategy 34:41 MS(n) mass spectrometry strategy 10:41 MS(n) mass spectrometry strategy for carbohydrate analysis 10:67 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 2 144 theme quadrupole 192:201 arg1 spectrometer 217:228 a quadrupole ion-trap mass spectrometer 190:228 a quadrupole ion-trap mass spectrometer 190:228 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 3 145 theme Y-type 334:339 arg1 fragmentation 341:353 Y-type fragmentation 334:353 Y-type fragmentation 334:353 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 3 145 theme Y-type 334:339 arg1 B 332:332 B 332:332 B 332:332 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 10 146 theme haptenic 1782:1789 arg1 [6,7 1791:1794 unique haptenic [6,7] 1775:1795 unique haptenic [6,7] 1775:1795 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 7 147 dep linear 921:926 arg1 branched 929:936 branched 929:936 branched 929:936 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 0 148 theme mass 16:19 arg1 strategy 34:41 MS(n) mass spectrometry strategy 10:41 MS(n) mass spectrometry strategy for carbohydrate analysis 10:67 Enhancing MS(n) mass spectrometry strategy for carbohydrate analysis: A b2 ion spectral library. 25175058 18 149 theme ionization 2994:3003 arg1 modes 2985:2989 modes 2985:2989 modes of ionization and collisional activation 2985:3030 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 14 150 theme monomer 2422:2428 arg1 ID 2430:2431 monomer ID 2422:2431 monomer ID 2422:2431 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 4 151 theme " 494:494 arg1 Fragmentation 469:481 Fragmentation 469:481 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, 469:599 Fragmentation of "B2 ions" corresponding to glycosidic linkages such as Hex-Fuc, Hex-Hex, Hex-HexNAc, HexNAc-Hex and HexNAc-HexNAc, were systematically studied in low energy CID and collected to form a "B2 library". 25175058 8 152 theme improved 1283:1290 arg1 tool 1292:1295 an improved tool 1280:1295 an improved tool for glycomics 1280:1309 This MS strategy for linkage determination of carbohydrates aided by a "B2 library" was developed with a scope for expansion, providing an improved tool for glycomics. 25175058 3 153 theme glycosidic 420:429 arg1 linkages 431:438 specific glycosidic linkages 411:438 specific glycosidic linkages using progressive MS stages 411:466 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 6 154 theme " 804:804 arg1 Patterns 784:791 Patterns 784:791 Patterns of "B2 ions" 784:804 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 3 155 theme B2 369:370 arg1 ions 372:375 disaccharide B2 ions 356:375 disaccharide B2 ions 356:375 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 7 156 from problem 1051:1057 arg1 practice 1062:1069 practice 1062:1069 practice 1062:1069 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 25175058 17 157 dep Science 2721:2727 arg1 http 2743:2746 http 2743:2746 http://glyco.nas.edu 2743:2762 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 17 157 dep Science 2721:2727 arg1 report 2735:2740 report 2735:2740 report 2735:2740 The National Academy of Science (NAS) report (http://glyco.nas.edu) recommends the need of more than 10,000 synthetic structures of carbohydrates to advance the field of glycomics. 25175058 10 158 theme antigenic 1800:1808 arg1 markers 1810:1816 antigenic markers 1800:1816 antigenic markers for oncology [8-11] and other applications 1800:1859 BIOLOGICAL SIGNIFICANCE Glycosylation is an important class of the "postranslationome", which includes manifold aspects of post-translational protein modification, affecting protein conformation, providing ligands for protein receptors [1-5], and encoding unique haptenic [6,7] or antigenic markers for oncology [8-11] and other applications. 25175058 14 159 theme activation 2332:2341 arg1 stages 2312:2317 sequential stages 2301:2317 sequential stages of collision activation (MS(n)) 2301:2349 Mass spectra from sequential stages of collision activation (MS(n)) can supply structural details for precise characterization of linkage, monomer ID, substitutions, anomerity and branching [21-25]. 25175058 2 160 theme positive 310:317 arg1 mode 323:326 the positive ion mode 306:326 the positive ion mode 306:326 Using a quadrupole ion-trap mass spectrometer, successive fragmentations of sodiated oligosaccharides were carried out in the positive ion mode. 25175058 6 161 theme B2 797:798 arg1 ions 800:803 B2 ions 797:803 "B2 ions" 796:804 Patterns of "B2 ions" rely on relative stability of glycosidic bonds and carbohydrate-metal complexes in the gas phase. 25175058 13 162 theme expressed 2242:2250 arg1 transferases 2261:2272 the expressed glycosyl transferases [13-20] 2238:2280 the expressed glycosyl transferases [13-20] 2238:2280 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 9 163 from activities 1367:1376 arg1 lines 1422:1426 cancer cell lines 1410:1426 cancer cell lines 1410:1426 We validated this method examining levels of expressed activities of two glycosyl transferases in cancer cell lines: β3(B3GALNT2) and β4GalNAcT(B4GALNT3&4) that generate GalNAcβ3GlcNAcβ and GalNAcβ4GlcNAcβ. 25175058 11 164 theme ring 1918:1921 arg1 size 1923:1926 ring size 1918:1926 ring size 1918:1926 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 11 164 theme ring 1918:1921 arg1 units 1901:1905 individual monomeric units 1880:1905 individual monomeric units 1880:1905 Identification of individual monomeric units, linkages, ring size, branching and anomerity has posed significant challenges to mass spectrometrists. 25175058 3 165 theme MS 458:459 arg1 stages 461:466 progressive MS stages 446:466 progressive MS stages 446:466 In B,Y-type fragmentation, disaccharide B2 ions are generated which correspond to specific glycosidic linkages using progressive MS stages. 25175058 13 166 theme biological 2209:2218 arg1 system 2220:2225 the biological system 2205:2225 the biological system 2205:2225 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 18 167 theme MS 3051:3052 arg1 library 3057:3063 an MS(n) library 3048:3063 an MS(n) library for carbohydrate identification 3048:3095 This study demonstrates that the general reproducibility of ion trap spectra, and energy independence from modes of ionization and collisional activation, make compiling an MS(n) library for carbohydrate identification an achievable research target [26]. 25175058 13 168 theme likely 2166:2171 arg1 possibilities 2173:2185 likely possibilities 2166:2185 likely possibilities 2166:2185 While the potential isomers in oligosaccharides are impossibly large [12], likely possibilities can be limited by the biological system, including the expressed glycosyl transferases [13-20]. 25175058 7 169 theme isomeric 1094:1101 arg1 sequence 1103:1110 an isomeric sequence 1091:1110 an isomeric sequence 1091:1110 MS(n) studies of linear, branched trisaccharides and tetrasaccharides show that isomers for which B2 ion information is not available are rarely a problem in practice by their absence in an isomeric sequence or by their scarcity in nature. 26083095 0 0 theme Chondroitin 81:91 arg1 Sulfate 93:99 Fucosylated Chondroitin Sulfate 69:99 Fucosylated Chondroitin Sulfate 69:99 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 3 1 from source 410:415 arg1 material 384:391 chondroitin raw material 368:391 chondroitin raw material from a microbial source 368:415 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 5 2 theme structural 961:970 arg1 point 972:976 a structural point 959:976 a structural point of view 959:984 A semisynthetic fCS was characterized from a structural point of view by means of 2D-NMR techniques, and preliminarily assayed in an anticoagulant test. 26083095 0 3 theme Fucosylated 69:79 arg1 Sulfate 93:99 Fucosylated Chondroitin Sulfate 69:99 Fucosylated Chondroitin Sulfate 69:99 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 4 4 theme fucosylated 598:608 arg1 polysaccharides 610:624 the fucosylated polysaccharides 594:624 the fucosylated polysaccharides 594:624 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 3 5 from chemo- 421:426 arg1 work 362:365 this work 357:365 this work 357:365 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 6 theme fucosylated 725:735 arg1 sulfates 749:756 sourced fucosylated chondroitin sulfates 717:756 sourced fucosylated chondroitin sulfates (fCSs) 717:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 6 theme fucosylated 725:735 arg1 fCSs 759:762 fCSs 759:762 fCSs 759:762 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 2 7 theme fine 302:305 arg1 control 307:313 a fine control 300:313 a fine control of the obtained structural parameters 300:351 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 4 8 theme interesting 881:891 arg1 activities 904:913 several very interesting biological activities 868:913 several very interesting biological activities 868:913 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 3 9 theme chondroitin 368:378 arg1 material 384:391 chondroitin raw material 368:391 chondroitin raw material from a microbial source 368:415 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 10 theme sea 814:816 arg1 cucumbers 818:826 sea cucumbers 814:826 sea cucumbers (Echinoidea, Holothuroidea) 814:854 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 10 theme sea 814:816 arg1 Holothuroidea 841:853 Holothuroidea 841:853 Holothuroidea 841:853 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 1 11 gly O-glycosylation 111:125 arg1 polysaccharides 130:144 polysaccharides 130:144 polysaccharides 130:144 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 3 12 from acceptors 532:540 arg1 reactions 558:566 fucosylation reactions 545:566 fucosylation reactions 545:566 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 5 13 theme semisynthetic 918:930 arg1 fCS 932:934 A semisynthetic fCS 916:934 A semisynthetic fCS 916:934 A semisynthetic fCS was characterized from a structural point of view by means of 2D-NMR techniques, and preliminarily assayed in an anticoagulant test. 26083095 4 14 theme biological 893:902 arg1 activities 904:913 several very interesting biological activities 868:913 several very interesting biological activities 868:913 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 15 gly fucosylated 598:608 arg1 polysaccharides 610:624 the fucosylated polysaccharides 594:624 the fucosylated polysaccharides 594:624 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 3 16 theme fucosylation 545:556 arg1 reactions 558:566 fucosylation reactions 545:566 fucosylation reactions 545:566 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 17 theme first 696:700 arg1 time 702:705 the first time 692:705 the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs) 692:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 5 18 theme anticoagulant 1049:1061 arg1 test 1063:1066 an anticoagulant test 1046:1066 an anticoagulant test 1046:1066 A semisynthetic fCS was characterized from a structural point of view by means of 2D-NMR techniques, and preliminarily assayed in an anticoagulant test. 26083095 2 19 theme obtained 322:329 arg1 parameters 342:351 the obtained structural parameters 318:351 the obtained structural parameters 318:351 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 4 20 theme sourced 717:723 arg1 sulfates 749:756 sourced fucosylated chondroitin sulfates 717:756 sourced fucosylated chondroitin sulfates (fCSs) 717:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 20 theme sourced 717:723 arg1 fCSs 759:762 fCSs 759:762 fCSs 759:762 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 0 21 theme Chemical 0:7 arg1 Fucosylation 9:20 Chemical Fucosylation 0:20 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 0:100 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 5 22 theme 2D-NMR 998:1003 arg1 techniques 1005:1014 2D-NMR techniques 998:1014 2D-NMR techniques 998:1014 A semisynthetic fCS was characterized from a structural point of view by means of 2D-NMR techniques, and preliminarily assayed in an anticoagulant test. 26083095 4 23 theme nonanimal 707:715 arg1 time 702:705 the first time 692:705 the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs) 692:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 3 24 theme raw 380:382 arg1 material 384:391 chondroitin raw material 368:391 chondroitin raw material from a microbial source 368:415 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 25 theme Further 569:575 arg1 manipulations 577:589 Further manipulations 569:589 Further manipulations on the fucosylated polysaccharides 569:624 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 1 26 theme Chemical 102:109 arg1 reaction 170:177 an almost unexplored reaction 149:177 an almost unexplored reaction 149:177 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 1 26 theme Chemical 102:109 arg1 O-glycosylation 111:125 Chemical O-glycosylation 102:125 Chemical O-glycosylation of polysaccharides 102:144 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 3 27 theme polysaccharide 471:484 arg1 acceptors 532:540 glycosyl acceptors 523:540 glycosyl acceptors in fucosylation reactions 523:566 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 3 27 theme polysaccharide 471:484 arg1 intermediates 486:498 two polysaccharide intermediates 467:498 two polysaccharide intermediates 467:498 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 28 theme chondroitin 737:747 arg1 sulfates 749:756 sourced fucosylated chondroitin sulfates 717:756 sourced fucosylated chondroitin sulfates (fCSs) 717:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 28 theme chondroitin 737:747 arg1 fCSs 759:762 fCSs 759:762 fCSs 759:762 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 5 29 theme view 981:984 arg1 point 972:976 a structural point 959:976 a structural point of view 959:984 A semisynthetic fCS was characterized from a structural point of view by means of 2D-NMR techniques, and preliminarily assayed in an anticoagulant test. 26083095 3 30 from work 362:365 arg1 chemo- 421:426 chemo- 421:426 chemo- 421:426 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 31 theme several 868:874 arg1 activities 904:913 several very interesting biological activities 868:913 several very interesting biological activities 868:913 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 0 32 theme Polysaccharide 27:40 arg1 Fucosylation 9:20 Chemical Fucosylation 0:20 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 0:100 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 0 33 gly Fucosylation 9:20 arg1 Polysaccharide 27:40 a Polysaccharide 25:40 a Polysaccharide 25:40 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 4 34 from manipulations 577:589 arg1 polysaccharides 610:624 the fucosylated polysaccharides 594:624 the fucosylated polysaccharides 594:624 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 0 35 theme Semisynthetic 45:57 arg1 Access 59:64 A Semisynthetic Access 43:64 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 0:100 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 4 36 gly fucosylated 725:735 arg1 sulfates 749:756 sourced fucosylated chondroitin sulfates 717:756 sourced fucosylated chondroitin sulfates (fCSs) 717:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 36 gly fucosylated 725:735 arg1 fCSs 759:762 fCSs 759:762 fCSs 759:762 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 2 37 theme such 235:238 arg1 biomacromolecules 248:264 such complex biomacromolecules 235:264 such complex biomacromolecules 235:264 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 1 38 theme polysaccharides 130:144 arg1 reaction 170:177 an almost unexplored reaction 149:177 an almost unexplored reaction 149:177 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 1 38 theme polysaccharides 130:144 arg1 O-glycosylation 111:125 Chemical O-glycosylation 102:125 Chemical O-glycosylation of polysaccharides 102:144 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 2 39 theme complex 240:246 arg1 biomacromolecules 248:264 such complex biomacromolecules 235:264 such complex biomacromolecules 235:264 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 2 40 theme parameters 342:351 arg1 control 307:313 a fine control 300:313 a fine control of the obtained structural parameters 300:351 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 2 41 theme quantitative 271:282 arg1 manner 284:289 a quantitative manner 269:289 a quantitative manner 269:289 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 2 42 theme structural 331:340 arg1 parameters 342:351 the obtained structural parameters 318:351 the obtained structural parameters 318:351 This is mainly due to the difficulties in derivatizing such complex biomacromolecules in a quantitative manner and with a fine control of the obtained structural parameters. 26083095 4 43 theme multiple 637:644 arg1 de-O-benzylation 646:661 multiple de-O-benzylation 637:661 multiple de-O-benzylation 637:661 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 43 theme multiple 637:644 arg1 -polysaccharides 764:779 furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides 678:779 furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) 678:854 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 0 44 dep Fucosylation 9:20 arg1 Access 59:64 A Semisynthetic Access 43:64 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 0:100 Chemical Fucosylation of a Polysaccharide: A Semisynthetic Access to Fucosylated Chondroitin Sulfate. 26083095 4 45 theme furnished 678:686 arg1 de-O-benzylation 646:661 multiple de-O-benzylation 637:661 multiple de-O-benzylation 637:661 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 45 theme furnished 678:686 arg1 -polysaccharides 764:779 furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides 678:779 furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) 678:854 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 3 46 theme glycosyl 523:530 arg1 intermediates 486:498 two polysaccharide intermediates 467:498 two polysaccharide intermediates 467:498 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 3 46 theme glycosyl 523:530 arg1 acceptors 532:540 glycosyl acceptors 523:540 glycosyl acceptors in fucosylation reactions 523:566 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 3 47 theme microbial 400:408 arg1 source 410:415 a microbial source 398:415 a microbial source 398:415 In this work, chondroitin raw material from a microbial source was chemo- and regioselectively protected to give two polysaccharide intermediates, that acted in turn as glycosyl acceptors in fucosylation reactions. 26083095 4 48 dep nonanimal 707:715 arg1 sulfates 749:756 sourced fucosylated chondroitin sulfates 717:756 sourced fucosylated chondroitin sulfates (fCSs) 717:763 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 4 48 dep nonanimal 707:715 arg1 fCSs 759:762 fCSs 759:762 fCSs 759:762 Further manipulations on the fucosylated polysaccharides, including multiple de-O-benzylation and sulfation, furnished for the first time nonanimal sourced fucosylated chondroitin sulfates (fCSs)-polysaccharides obtained so far exclusively from sea cucumbers (Echinoidea, Holothuroidea) and showing several very interesting biological activities. 26083095 1 49 theme unexplored 159:168 arg1 reaction 170:177 an almost unexplored reaction 149:177 an almost unexplored reaction 149:177 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26083095 1 49 theme unexplored 159:168 arg1 O-glycosylation 111:125 Chemical O-glycosylation 102:125 Chemical O-glycosylation of polysaccharides 102:144 Chemical O-glycosylation of polysaccharides is an almost unexplored reaction. 26507102 9 0 gly monosialylated 1217:1230 arg1 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 10 1 theme normal 1413:1418 arg1 % 1438:1438 26.7 ± 6.7% 1428:1438 26.7 ± 6.7% 1428:1438 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 10 1 theme normal 1413:1418 arg1 joints 1420:1425 normal joints 1413:1425 normal joints (26.7 ± 6.7%) 1413:1439 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 7 2 theme western 950:956 arg1 blot 958:961 western blot 950:961 western blot 950:961 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 11 3 theme C-terminal 1568:1577 arg1 site 1600:1603 A C-terminal proteolytic cleavage site 1566:1603 A C-terminal proteolytic cleavage site in lubricin 1566:1615 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 14 4 theme superficial 2181:2191 arg1 layer 2193:2197 the superficial layer 2177:2197 the superficial layer 2177:2197 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 11 5 theme synovial 1630:1637 arg1 fluid 1639:1643 synovial fluid 1630:1643 synovial fluid from normal and osteochondral fragment joints 1630:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 11 6 from explants 1772:1779 arg1 media 1698:1702 media 1698:1702 media from interleukin-1β stimulated and unstimulated articular cartilage explants 1698:1779 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 10 7 dep = 1557:1557 arg1 % 1552:1552 21.2 ± 4.9% 1542:1552 21.2 ± 4.9% 1542:1552 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 10 8 theme ± 1433:1433 arg1 % 1438:1438 26.7 ± 6.7% 1428:1438 26.7 ± 6.7% 1428:1438 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 10 8 theme ± 1433:1433 arg1 joints 1420:1425 normal joints 1413:1425 normal joints (26.7 ± 6.7%) 1413:1439 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 1 9 theme articular 181:189 arg1 surface 201:207 the articular cartilage surface 177:207 the articular cartilage surface 177:207 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 14 10 theme lubricating 2154:2164 arg1 ability 2166:2172 the boundary lubricating ability 2141:2172 the boundary lubricating ability of the superficial layer of articular cartilage 2141:2220 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 14 11 theme reduced 2061:2067 arg1 sialation 2069:2077 The reduced sialation 2057:2077 The reduced sialation of lubricin in synovial fluid from diseased joints 2057:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 9 12 theme ± 1296:1296 arg1 % 1301:1301 53.8 ± 7.8% 1291:1301 53.8 ± 7.8% 1291:1301 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 1 13 theme surface 201:207 arg1 lubrication 162:172 the boundary lubrication 149:172 the boundary lubrication of the articular cartilage surface 149:207 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 12 14 from profile 1859:1865 arg1 fluid 1891:1895 synovial fluid 1882:1895 synovial fluid from diseased equine joints 1882:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 14 from profile 1859:1865 arg1 This 1794:1797 This 1794:1797 This 1794:1797 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 14 from profile 1859:1865 arg1 demonstration 1812:1824 the first demonstration 1802:1824 the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1802:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 8 15 theme Core 1096:1099 arg1 O-glycans 1103:1111 Core 1 O-glycans predominating 1096:1125 Core 1 O-glycans predominating 1096:1125 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 1 16 gly glycoprotein 112:123 arg1 glycoprotein 112:123 The glycoprotein lubricin 108:132 The glycoprotein lubricin 108:132 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 7 17 theme 3-day 892:896 arg1 intervals 898:906 3-day intervals 892:906 3-day intervals 892:906 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 5 18 with lesions 670:676 arg1 cartilage 630:638 normal articular cartilage 613:638 normal articular cartilage 613:638 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 5 19 dep METHODS 543:549 arg1 samples 566:572 Synovial fluid samples 551:572 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 543:774 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 5 19 dep METHODS 543:549 arg1 analysed 726:733 analysed 726:733 were analysed for the lubricin glycosylation profiles 721:773 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 14 20 theme diseased 2114:2121 arg1 joints 2123:2128 diseased joints 2114:2128 diseased joints 2114:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 10 21 dep fragments 1531:1539 arg1 = 1557:1557 = 1557:1557 = 1557:1557 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 3 22 from profile 351:357 arg1 fluid 383:387 synovial fluid 374:387 synovial fluid from horses with osteoarthritis 374:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 3 22 from profile 351:357 arg1 horses 394:399 horses 394:399 horses with osteoarthritis 394:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 5 23 theme lubricin 743:750 arg1 profiles 766:773 the lubricin glycosylation profiles 739:773 the lubricin glycosylation profiles 739:773 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 5 24 theme Synovial 551:558 arg1 samples 566:572 Synovial fluid samples 551:572 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 543:774 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 3 25 theme lubricin 362:369 arg1 profile 351:357 the glycosylation profile 333:357 the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis 333:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 9 26 theme Synovial 1128:1135 arg1 fluid 1137:1141 Synovial fluid 1128:1141 Synovial fluid from normal joints (49.5 ± 1.9%) 1128:1174 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 13 27 theme proteolytic 1992:2002 arg1 site 2013:2016 an identical proteolytic cleavage site 1979:2016 an identical proteolytic cleavage site of lubricin 1979:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 13 27 theme proteolytic 1992:2002 arg1 lubricin 2021:2028 lubricin 2021:2028 lubricin 2021:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 3 28 from secretion 431:439 arg1 model 507:511 an in vitro inflammation cartilage model 472:511 an in vitro inflammation cartilage model 472:511 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 2 29 theme superficial 257:267 arg1 layer 269:273 the superficial layer 253:273 the superficial layer where lubricin is synthesised 253:303 The early events of osteoarthritis involve the superficial layer where lubricin is synthesised. 26507102 1 30 theme boundary 153:160 arg1 lubrication 162:172 the boundary lubrication 149:172 the boundary lubrication of the articular cartilage surface 149:207 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 10 31 with joints 1505:1510 arg1 fragments 1531:1539 osteochondral fragments 1517:1539 osteochondral fragments (21.2 ± 4.9%, P = 0.03) 1517:1563 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 11 32 theme articular 1752:1760 arg1 explants 1772:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 3 33 from degradation 445:455 arg1 model 507:511 an in vitro inflammation cartilage model 472:511 an in vitro inflammation cartilage model 472:511 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 5 34 with joints 601:606 arg1 cartilage 630:638 normal articular cartilage 613:638 normal articular cartilage 613:638 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 0 35 from Characterisation 0:15 arg1 fluid 41:45 synovial fluid 32:45 synovial fluid from horses with osteoarthritis 32:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 0 35 from Characterisation 0:15 arg1 horses 52:57 horses 52:57 horses with osteoarthritis 52:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 14 36 theme synovial 2094:2101 arg1 fluid 2103:2107 synovial fluid 2094:2107 synovial fluid from diseased joints 2094:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 12 37 theme equine 1911:1916 arg1 joints 1918:1923 diseased equine joints 1902:1923 diseased equine joints 1902:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 5 38 theme osteoarthritic 655:668 arg1 lesions 670:676 structural osteoarthritic lesions 644:676 structural osteoarthritic lesions 644:676 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 5 39 with horses 589:594 arg1 joints 601:606 joints 601:606 joints with normal articular cartilage 601:638 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 5 39 with horses 589:594 arg1 lesions 670:676 structural osteoarthritic lesions 644:676 structural osteoarthritic lesions 644:676 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 3 40 theme synovial 374:381 arg1 fluid 383:387 synovial fluid 374:387 synovial fluid from horses with osteoarthritis 374:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 10 41 theme disialylated 1466:1477 arg1 O-glycan 1486:1493 disialylated Core 1 O-glycan 1466:1493 disialylated Core 1 O-glycan 1466:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 9 42 with joints 1317:1322 arg1 fragments 1343:1351 osteochondral fragments 1329:1351 osteochondral fragments (57.3 ± 8.8%, P = 0.001) 1329:1376 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 42 with joints 1317:1322 arg1 osteoarthritis 1275:1288 osteoarthritis 1275:1288 osteoarthritis (53.8 ± 7.8%, P = 0.03) 1275:1312 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 43 theme lubricin 1870:1877 arg1 profile 1859:1865 the glycosylation profile 1841:1865 the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1841:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 3 44 theme lubricin 460:467 arg1 degradation 445:455 degradation 445:455 degradation 445:455 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 3 44 theme lubricin 460:467 arg1 secretion 431:439 secretion 431:439 secretion 431:439 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 12 45 theme synovial 1882:1889 arg1 fluid 1891:1895 synovial fluid 1882:1895 synovial fluid from diseased equine joints 1882:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 5 46 theme articular 620:628 arg1 cartilage 630:638 normal articular cartilage 613:638 normal articular cartilage 613:638 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 14 47 from events 2252:2257 arg1 progression 2266:2276 the progression 2262:2276 the progression of osteoarthritis 2262:2294 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 3 48 theme cartilage 497:505 arg1 model 507:511 an in vitro inflammation cartilage model 472:511 an in vitro inflammation cartilage model 472:511 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 0 49 theme lubricin 20:27 arg1 Characterisation 0:15 Characterisation 0:15 Characterisation of lubricin in synovial fluid from horses with osteoarthritis 0:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 7 50 theme Media 865:869 arg1 samples 871:877 Media samples 865:877 Media samples collected at 3-day intervals 865:906 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 8 51 theme RESULTS 1002:1008 arg1 profiles 1026:1033 RESULTS O-glycosylation profiles 1002:1033 RESULTS O-glycosylation profiles in synovial fluid 1002:1051 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 11 52 from site 1600:1603 arg1 lubricin 1608:1615 lubricin 1608:1615 lubricin 1608:1615 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 11 53 theme osteochondral 1661:1673 arg1 joints 1684:1689 normal and osteochondral fragment joints 1650:1689 normal and osteochondral fragment joints 1650:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 11 54 located found 1621:1625 arg2 site 1600:1603 A C-terminal proteolytic cleavage site 1566:1603 A C-terminal proteolytic cleavage site in lubricin 1566:1615 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 11 54 located found 1621:1625 arg1 fluid 1639:1643 synovial fluid 1630:1643 synovial fluid from normal and osteochondral fragment joints 1630:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 0 55 theme synovial 32:39 arg1 fluid 41:45 synovial fluid 32:45 synovial fluid from horses with osteoarthritis 32:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 10 56 theme osteochondral 1517:1529 arg1 fragments 1531:1539 osteochondral fragments 1517:1539 osteochondral fragments (21.2 ± 4.9%, P = 0.03) 1517:1563 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 7 57 link enzyme-linked 967:979 arg1 assay 995:999 enzyme-linked immunosorbent assay 967:999 enzyme-linked immunosorbent assay 967:999 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 3 58 dep in 475:476 arg1 vitro 478:482 vitro 478:482 vitro 478:482 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 8 59 theme synovial 1038:1045 arg1 fluid 1047:1051 synovial fluid 1038:1051 synovial fluid 1038:1051 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 2 60 theme early 214:218 arg1 events 220:225 The early events 210:225 The early events of osteoarthritis 210:243 The early events of osteoarthritis involve the superficial layer where lubricin is synthesised. 26507102 9 61 theme monosialylated 1217:1230 arg1 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 62 theme change 1831:1836 arg1 This 1794:1797 This 1794:1797 This 1794:1797 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 62 theme change 1831:1836 arg1 demonstration 1812:1824 the first demonstration 1802:1824 the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1802:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 13 63 theme cleavage 2004:2011 arg1 site 2013:2016 an identical proteolytic cleavage site 1979:2016 an identical proteolytic cleavage site of lubricin 1979:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 13 63 theme cleavage 2004:2011 arg1 lubricin 2021:2028 lubricin 2021:2028 lubricin 2021:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 9 64 dep = 1306:1306 arg1 % 1301:1301 53.8 ± 7.8% 1291:1301 53.8 ± 7.8% 1291:1301 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 10 65 theme ± 1547:1547 arg1 % 1552:1552 21.2 ± 4.9% 1542:1552 21.2 ± 4.9% 1542:1552 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 11 66 theme cleavage 1591:1598 arg1 site 1600:1603 A C-terminal proteolytic cleavage site 1566:1603 A C-terminal proteolytic cleavage site in lubricin 1566:1615 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 10 67 from joints 1420:1425 arg1 fluid 1402:1406 synovial fluid 1393:1406 synovial fluid from normal joints (26.7 ± 6.7%) 1393:1439 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 14 68 theme articular 2202:2210 arg1 cartilage 2212:2220 articular cartilage 2202:2220 articular cartilage 2202:2220 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 5 69 theme osteochondral 696:708 arg1 fragments 710:718 osteochondral fragments 696:718 osteochondral fragments 696:718 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 6 70 theme cartilage 786:794 arg1 explants 796:803 Articular cartilage explants 776:803 Articular cartilage explants 776:803 Articular cartilage explants were stimulated with or without interleukin-1β for 25 days. 26507102 7 71 theme enzyme-linked 967:979 arg1 assay 995:999 enzyme-linked immunosorbent assay 967:999 enzyme-linked immunosorbent assay 967:999 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 11 72 theme normal 1650:1655 arg1 joints 1684:1689 normal and osteochondral fragment joints 1650:1689 normal and osteochondral fragment joints 1650:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 12 73 theme diseased 1902:1909 arg1 joints 1918:1923 diseased equine joints 1902:1923 diseased equine joints 1902:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 14 74 theme layer 2193:2197 arg1 ability 2166:2172 the boundary lubricating ability 2141:2172 the boundary lubricating ability of the superficial layer of articular cartilage 2141:2220 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 14 75 from sialation 2069:2077 arg1 fluid 2103:2107 synovial fluid 2094:2107 synovial fluid from diseased joints 2094:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 14 75 from sialation 2069:2077 arg1 joints 2123:2128 diseased joints 2114:2128 diseased joints 2114:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 10 76 theme synovial 1393:1400 arg1 fluid 1402:1406 synovial fluid 1393:1406 synovial fluid from normal joints (26.7 ± 6.7%) 1393:1439 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 7 77 theme quantitative 925:936 arg1 proteomics 938:947 quantitative proteomics 925:947 quantitative proteomics 925:947 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 1 78 theme cartilage 191:199 arg1 surface 201:207 the articular cartilage surface 177:207 the articular cartilage surface 177:207 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 12 79 theme normal 1949:1954 arg1 joints 1956:1961 normal joints 1949:1961 normal joints 1949:1961 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 14 80 theme boundary 2145:2152 arg1 ability 2166:2172 the boundary lubricating ability 2141:2172 the boundary lubricating ability of the superficial layer of articular cartilage 2141:2220 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 5 81 theme glycosylation 752:764 arg1 profiles 766:773 the lubricin glycosylation profiles 739:773 the lubricin glycosylation profiles 739:773 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 11 82 theme proteolytic 1579:1589 arg1 site 1600:1603 A C-terminal proteolytic cleavage site 1566:1603 A C-terminal proteolytic cleavage site in lubricin 1566:1615 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 14 83 from joints 2123:2128 arg1 fluid 2103:2107 synovial fluid 2094:2107 synovial fluid from diseased joints 2094:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 14 83 from joints 2123:2128 arg1 sialation 2069:2077 The reduced sialation 2057:2077 The reduced sialation of lubricin in synovial fluid from diseased joints 2057:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 3 84 from horses 394:399 arg1 fluid 383:387 synovial fluid 374:387 synovial fluid from horses with osteoarthritis 374:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 3 84 from horses 394:399 arg1 profile 351:357 the glycosylation profile 333:357 the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis 333:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 2 85 theme osteoarthritis 230:243 arg1 events 220:225 The early events 210:225 The early events of osteoarthritis 210:243 The early events of osteoarthritis involve the superficial layer where lubricin is synthesised. 26507102 1 86 theme glycoprotein 112:123 arg1 lubricin 125:132 The glycoprotein lubricin 108:132 The glycoprotein lubricin 108:132 REASON FOR PERFORMING STUDY The glycoprotein lubricin contributes to the boundary lubrication of the articular cartilage surface. 26507102 3 87 theme glycosylation 337:349 arg1 profile 351:357 the glycosylation profile 333:357 the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis 333:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 3 88 with horses 394:399 arg1 osteoarthritis 406:419 osteoarthritis 406:419 osteoarthritis 406:419 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 9 89 dep osteoarthritis 1275:1288 arg1 = 1306:1306 = 1306:1306 = 1306:1306 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 90 from joints 1918:1923 arg1 lubricin 1870:1877 lubricin 1870:1877 lubricin in synovial fluid from diseased equine joints 1870:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 90 from joints 1918:1923 arg1 fluid 1891:1895 synovial fluid 1882:1895 synovial fluid from diseased equine joints 1882:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 13 91 theme lubricin 2021:2028 arg1 site 2013:2016 an identical proteolytic cleavage site 1979:2016 an identical proteolytic cleavage site of lubricin 1979:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 13 91 theme lubricin 2021:2028 arg1 lubricin 2021:2028 lubricin 2021:2028 lubricin 2021:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 11 92 theme stimulated 1724:1733 arg1 explants 1772:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 5 93 theme fluid 560:564 arg1 samples 566:572 Synovial fluid samples 551:572 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 543:774 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 11 94 from joints 1684:1689 arg1 fluid 1639:1643 synovial fluid 1630:1643 synovial fluid from normal and osteochondral fragment joints 1630:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 9 95 dep = 1369:1369 arg1 % 1364:1364 57.3 ± 8.8% 1354:1364 57.3 ± 8.8% 1354:1364 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 14 96 theme osteoarthritis 2281:2294 arg1 progression 2266:2276 the progression 2262:2276 the progression of osteoarthritis 2262:2294 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 9 97 theme osteochondral 1329:1341 arg1 fragments 1343:1351 osteochondral fragments 1329:1351 osteochondral fragments (57.3 ± 8.8%, P = 0.001) 1329:1376 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 8 98 theme predominating 1113:1125 arg1 O-glycans 1103:1111 Core 1 O-glycans predominating 1096:1125 Core 1 O-glycans predominating 1096:1125 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 0 99 from horses 52:57 arg1 fluid 41:45 synovial fluid 32:45 synovial fluid from horses with osteoarthritis 32:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 0 99 from horses 52:57 arg1 Characterisation 0:15 Characterisation 0:15 Characterisation of lubricin in synovial fluid from horses with osteoarthritis 0:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 12 100 from fluid 1891:1895 arg1 profile 1859:1865 the glycosylation profile 1841:1865 the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1841:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 9 101 theme normal 1148:1153 arg1 % 1173:1173 49.5 ± 1.9% 1163:1173 49.5 ± 1.9% 1163:1173 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 101 theme normal 1148:1153 arg1 joints 1155:1160 normal joints 1148:1160 normal joints (49.5 ± 1.9%) 1148:1174 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 11 102 theme cartilage 1762:1770 arg1 explants 1772:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 5 103 theme structural 644:653 arg1 lesions 670:676 structural osteoarthritic lesions 644:676 structural osteoarthritic lesions 644:676 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 0 104 with horses 52:57 arg1 osteoarthritis 64:77 osteoarthritis 64:77 osteoarthritis 64:77 Characterisation of lubricin in synovial fluid from horses with osteoarthritis. 26507102 4 105 theme STUDY 514:518 arg1 DESIGN 520:525 STUDY DESIGN 514:525 STUDY DESIGN 514:525 STUDY DESIGN In vitro study. 26507102 9 106 from joints 1155:1160 arg1 fluid 1137:1141 Synovial fluid 1128:1141 Synovial fluid from normal joints (49.5 ± 1.9%) 1128:1174 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 6 107 theme Articular 776:784 arg1 explants 796:803 Articular cartilage explants 776:803 Articular cartilage explants 776:803 Articular cartilage explants were stimulated with or without interleukin-1β for 25 days. 26507102 11 108 theme unstimulated 1739:1750 arg1 explants 1772:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 interleukin-1β stimulated and unstimulated articular cartilage explants 1709:1779 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 9 109 contain contained 1176:1184 arg2 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 109 contain contained 1176:1184 arg2 amounts 1206:1212 significantly lower amounts 1186:1212 significantly lower amounts of monosialylated Core 1 O-glycans 1186:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 109 contain contained 1176:1184 arg1 fluid 1137:1141 Synovial fluid 1128:1141 Synovial fluid from normal joints (49.5 ± 1.9%) 1128:1174 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 110 from lubricin 1870:1877 arg1 fluid 1891:1895 synovial fluid 1882:1895 synovial fluid from diseased equine joints 1882:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 110 from lubricin 1870:1877 arg1 joints 1918:1923 diseased equine joints 1902:1923 diseased equine joints 1902:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 9 111 theme ± 1168:1168 arg1 % 1173:1173 49.5 ± 1.9% 1163:1173 49.5 ± 1.9% 1163:1173 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 111 theme ± 1168:1168 arg1 joints 1155:1160 normal joints 1148:1160 normal joints (49.5 ± 1.9%) 1148:1174 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 3 112 theme in 475:476 arg1 model 507:511 an in vitro inflammation cartilage model 472:511 an in vitro inflammation cartilage model 472:511 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 5 113 theme normal 613:618 arg1 cartilage 630:638 normal articular cartilage 613:638 normal articular cartilage 613:638 METHODS Synovial fluid samples collected from horses with joints with normal articular cartilage and structural osteoarthritic lesions; with and without osteochondral fragments, were analysed for the lubricin glycosylation profiles. 26507102 8 114 from profiles 1026:1033 arg1 fluid 1047:1051 synovial fluid 1038:1051 synovial fluid 1038:1051 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 9 115 dep fragments 1343:1351 arg1 = 1369:1369 = 1369:1369 = 1369:1369 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 10 116 theme higher 1448:1453 arg1 O-glycan 1486:1493 disialylated Core 1 O-glycan 1466:1493 disialylated Core 1 O-glycan 1466:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 10 116 theme higher 1448:1453 arg1 amounts 1455:1461 higher amounts 1448:1461 higher amounts of disialylated Core 1 O-glycan 1448:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 14 117 theme lubricin 2082:2089 arg1 sialation 2069:2077 The reduced sialation 2057:2077 The reduced sialation of lubricin in synovial fluid from diseased joints 2057:2128 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 12 118 dep CONCLUSIONS 1782:1792 arg1 This 1794:1797 This 1794:1797 This 1794:1797 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 118 dep CONCLUSIONS 1782:1792 arg1 demonstration 1812:1824 the first demonstration 1802:1824 the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1802:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 119 theme glycosylation 1845:1857 arg1 profile 1859:1865 the glycosylation profile 1841:1865 the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1841:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 10 120 theme O-glycan 1486:1493 arg1 O-glycan 1486:1493 disialylated Core 1 O-glycan 1466:1493 disialylated Core 1 O-glycan 1466:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 10 120 theme O-glycan 1486:1493 arg1 amounts 1455:1461 higher amounts 1448:1461 higher amounts of disialylated Core 1 O-glycan 1448:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 12 121 gly glycosylation 1845:1857 arg1 lubricin 1870:1877 lubricin 1870:1877 lubricin in synovial fluid from diseased equine joints 1870:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 10 122 theme Core 1479:1482 arg1 O-glycan 1486:1493 disialylated Core 1 O-glycan 1466:1493 disialylated Core 1 O-glycan 1466:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 9 123 theme lower 1200:1204 arg1 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 123 theme lower 1200:1204 arg1 amounts 1206:1212 significantly lower amounts 1186:1212 significantly lower amounts of monosialylated Core 1 O-glycans 1186:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 3 124 theme inflammation 484:495 arg1 model 507:511 an in vitro inflammation cartilage model 472:511 an in vitro inflammation cartilage model 472:511 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 8 125 theme O-glycosylation 1010:1024 arg1 profiles 1026:1033 RESULTS O-glycosylation profiles 1002:1033 RESULTS O-glycosylation profiles in synovial fluid 1002:1051 RESULTS O-glycosylation profiles in synovial fluid revealed both Core 1 and 2 O-glycans, with Core 1 O-glycans predominating. 26507102 12 126 from demonstration 1812:1824 arg1 profile 1859:1865 the glycosylation profile 1841:1865 the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1841:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 13 127 theme identical 1982:1990 arg1 site 2013:2016 an identical proteolytic cleavage site 1979:2016 an identical proteolytic cleavage site of lubricin 1979:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 13 127 theme identical 1982:1990 arg1 lubricin 2021:2028 lubricin 2021:2028 lubricin 2021:2028 We demonstrate an identical proteolytic cleavage site of lubricin both in vitro and in vivo. 26507102 14 128 theme events 2252:2257 arg1 one 2235:2237 one 2235:2237 one 2235:2237 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 14 128 theme events 2252:2257 arg1 events 2252:2257 the early events 2242:2257 the early events in the progression of osteoarthritis 2242:2294 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 9 129 with joints 1263:1268 arg1 fragments 1343:1351 osteochondral fragments 1329:1351 osteochondral fragments (57.3 ± 8.8%, P = 0.001) 1329:1376 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 129 with joints 1263:1268 arg1 osteoarthritis 1275:1288 osteoarthritis 1275:1288 osteoarthritis (53.8 ± 7.8%, P = 0.03) 1275:1312 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 130 theme first 1806:1810 arg1 This 1794:1797 This 1794:1797 This 1794:1797 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 12 130 theme first 1806:1810 arg1 demonstration 1812:1824 the first demonstration 1802:1824 the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1802:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 26507102 10 131 gly disialylated 1466:1477 arg1 O-glycan 1486:1493 disialylated Core 1 O-glycan 1466:1493 disialylated Core 1 O-glycan 1466:1493 Additionally, synovial fluid from normal joints (26.7 ± 6.7%) showed higher amounts of disialylated Core 1 O-glycan than from joints with osteochondral fragments (21.2 ± 4.9%, P = 0.03). 26507102 11 132 theme fragment 1675:1682 arg1 joints 1684:1689 normal and osteochondral fragment joints 1650:1689 normal and osteochondral fragment joints 1650:1689 A C-terminal proteolytic cleavage site in lubricin was found in synovial fluid from normal and osteochondral fragment joints and in media from interleukin-1β stimulated and unstimulated articular cartilage explants. 26507102 9 133 theme Core 1232:1235 arg1 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 7 134 theme immunosorbent 981:993 arg1 assay 995:999 enzyme-linked immunosorbent assay 967:999 enzyme-linked immunosorbent assay 967:999 Media samples collected at 3-day intervals were analysed by quantitative proteomics, western blot and enzyme-linked immunosorbent assay. 26507102 14 135 theme early 2246:2250 arg1 events 2252:2257 the early events 2242:2257 the early events in the progression of osteoarthritis 2242:2294 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 3 136 gly glycosylation 337:349 arg1 lubricin 362:369 lubricin 362:369 lubricin 362:369 OBJECTIVES To characterise the glycosylation profile of lubricin in synovial fluid from horses with osteoarthritis and study secretion and degradation of lubricin in an in vitro inflammation cartilage model. 26507102 14 137 theme cartilage 2212:2220 arg1 ability 2166:2172 the boundary lubricating ability 2141:2172 the boundary lubricating ability of the superficial layer of articular cartilage 2141:2220 The reduced sialation of lubricin in synovial fluid from diseased joints may affect the boundary lubricating ability of the superficial layer of articular cartilage and could be one of the early events in the progression of osteoarthritis. 26507102 9 138 theme O-glycans 1239:1247 arg1 O-glycans 1239:1247 monosialylated Core 1 O-glycans 1217:1247 monosialylated Core 1 O-glycans 1217:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 9 138 theme O-glycans 1239:1247 arg1 amounts 1206:1212 significantly lower amounts 1186:1212 significantly lower amounts of monosialylated Core 1 O-glycans 1186:1247 Synovial fluid from normal joints (49.5 ± 1.9%) contained significantly lower amounts of monosialylated Core 1 O-glycans compared with joints with osteoarthritis (53.8 ± 7.8%, P = 0.03) or joints with osteochondral fragments (57.3 ± 8.8%, P = 0.001). 26507102 12 139 from change 1831:1836 arg1 profile 1859:1865 the glycosylation profile 1841:1865 the glycosylation profile of lubricin in synovial fluid from diseased equine joints 1841:1923 CONCLUSIONS This is the first demonstration of a change in the glycosylation profile of lubricin in synovial fluid from diseased equine joints compared with that from normal joints. 28594851 0 0 theme carcinoma 96:104 arg1 patients 106:113 oral squamous cell carcinoma patients 77:113 oral squamous cell carcinoma patients 77:113 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 12 1 theme serum 1727:1731 arg1 antibodies 1747:1756 antibodies 1747:1756 antibodies 1747:1756 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 12 1 theme serum 1727:1731 arg1 N-glycans 1733:1741 these serum N-glycans 1721:1741 these serum N-glycans 1721:1741 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 12 1 theme serum 1727:1731 arg1 candidates 1811:1820 the candidates 1807:1820 the candidates of potential biomarkers for OSCC diagnosis 1807:1863 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 10 2 theme serum 1447:1451 arg1 N-glycans 1453:1461 the high-molecular weight serum N-glycans 1421:1461 the high-molecular weight serum N-glycans 1421:1461 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 11 3 from levels 1657:1662 arg1 serum 1698:1702 patient serum 1690:1702 patient serum 1690:1702 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 0 4 theme cell 91:94 arg1 carcinoma 96:104 oral squamous cell carcinoma 77:104 oral squamous cell carcinoma patients 77:113 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 5 5 theme Total 705:709 arg1 proteins 717:724 Total serum proteins 705:724 Total serum proteins 705:724 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 10 6 theme high-molecular 1425:1438 arg1 N-glycans 1453:1461 the high-molecular weight serum N-glycans 1421:1461 the high-molecular weight serum N-glycans 1421:1461 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 1 7 theme important 187:195 arg1 role 197:200 important role 187:200 important role 187:200 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 7 8 theme serum 933:937 arg1 profiles 966:973 the serum anti-carbohydrate antibody profiles 929:973 the serum anti-carbohydrate antibody profiles 929:973 In addition, the serum anti-carbohydrate antibody profiles were also investigated by carbohydrate microarray. 28594851 5 9 theme alkylation 780:789 arg1 digestion 803:811 chemical reduction, alkylation and trypsin digestion 760:811 chemical reduction, alkylation and trypsin digestion 760:811 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 5 10 theme serum 711:715 arg1 proteins 717:724 Total serum proteins 705:724 Total serum proteins 705:724 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 9 11 theme fucosylation 1270:1281 arg1 degrees 1259:1265 varying degrees 1251:1265 varying degrees of fucosylation and sialylation 1251:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 8 12 theme OSCC 1123:1126 arg1 serum 1114:1118 serum 1114:1118 serum of OSCC 1114:1126 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 7 13 theme anti-carbohydrate 939:955 arg1 profiles 966:973 the serum anti-carbohydrate antibody profiles 929:973 the serum anti-carbohydrate antibody profiles 929:973 In addition, the serum anti-carbohydrate antibody profiles were also investigated by carbohydrate microarray. 28594851 4 14 theme squamous 646:653 arg1 carcinoma 660:668 oral squamous cell carcinoma 641:668 oral squamous cell carcinoma (OSCC) patients 641:684 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 4 14 theme squamous 646:653 arg1 OSCC 671:674 OSCC 671:674 OSCC 671:674 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 10 15 theme cutoff 1545:1550 arg1 values 1552:1557 the identified cutoff values 1530:1557 the identified cutoff values 1530:1557 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 8 16 theme N-glycans 1073:1081 arg1 abundances 1053:1062 the relative abundances 1040:1062 the relative abundances of seven N-glycans 1040:1081 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 2 17 theme common 306:311 arg1 phenomenon 313:322 a common phenomenon 304:322 a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis 304:405 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 2 17 theme common 306:311 arg1 Alteration 265:274 Alteration 265:274 Alteration of protein glycosylation 265:299 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 10 18 theme patients 1403:1410 arg1 patients 1403:1410 forty-eight OCCC patients 1386:1410 forty-eight OCCC patients 1386:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 10 18 theme patients 1403:1410 arg1 group 1377:1381 an independent validation group 1351:1381 an independent validation group of forty-eight OCCC patients 1351:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 4 19 theme oral 641:644 arg1 carcinoma 660:668 oral squamous cell carcinoma 641:668 oral squamous cell carcinoma (OSCC) patients 641:684 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 4 19 theme oral 641:644 arg1 OSCC 671:674 OSCC 671:674 OSCC 671:674 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 5 20 theme trypsin 795:801 arg1 digestion 803:811 chemical reduction, alkylation and trypsin digestion 760:811 chemical reduction, alkylation and trypsin digestion 760:811 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 8 21 theme diagnostic 1133:1142 arg1 accuracy 1144:1151 diagnostic accuracy 1133:1151 diagnostic accuracy greater than 75% 1133:1168 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 9 22 theme sialylation 1287:1297 arg1 degrees 1259:1265 varying degrees 1251:1265 varying degrees of fucosylation and sialylation 1251:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 12 23 theme biomarkers 1835:1844 arg1 antibodies 1747:1756 antibodies 1747:1756 antibodies 1747:1756 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 12 23 theme biomarkers 1835:1844 arg1 N-glycans 1733:1741 these serum N-glycans 1721:1741 these serum N-glycans 1721:1741 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 12 23 theme biomarkers 1835:1844 arg1 candidates 1811:1820 the candidates 1807:1820 the candidates of potential biomarkers for OSCC diagnosis 1807:1863 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 3 24 theme induced 450:456 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 24 theme induced 450:456 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 24 theme induced 450:456 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 4 25 from populations 625:635 arg1 N-glycomes 569:578 serum N-glycomes 563:578 serum N-glycomes 563:578 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 4 25 from populations 625:635 arg1 antibodies 602:611 anti-carbohydrate antibodies 584:611 anti-carbohydrate antibodies 584:611 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 10 26 theme identified 1534:1543 arg1 values 1552:1557 the identified cutoff values 1530:1557 the identified cutoff values 1530:1557 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 3 27 theme corresponding 458:470 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 27 theme corresponding 458:470 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 27 theme corresponding 458:470 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 0 28 theme N-glycome 6:14 arg1 characterization 16:31 Serum N-glycome characterization 0:31 Serum N-glycome characterization 0:31 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 11 29 dep elevated 1617:1624 arg1 accompanied 1626:1636 accompanied 1626:1636 accompanied with the decreased levels of nine IgG antibodies in patient serum 1626:1702 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 10 30 theme validation 1366:1375 arg1 patients 1403:1410 forty-eight OCCC patients 1386:1410 forty-eight OCCC patients 1386:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 10 30 theme validation 1366:1375 arg1 group 1377:1381 an independent validation group 1351:1381 an independent validation group of forty-eight OCCC patients 1351:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 1 31 theme protein 205:211 arg1 function 213:220 protein function 205:220 protein function 205:220 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 0 32 theme Serum 0:4 arg1 characterization 16:31 Serum N-glycome characterization 0:31 Serum N-glycome characterization 0:31 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 12 33 theme potential 1825:1833 arg1 biomarkers 1835:1844 potential biomarkers 1825:1844 potential biomarkers for OSCC diagnosis 1825:1863 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 7 34 theme carbohydrate 1001:1012 arg1 microarray 1014:1023 carbohydrate microarray 1001:1023 carbohydrate microarray 1001:1023 In addition, the serum anti-carbohydrate antibody profiles were also investigated by carbohydrate microarray. 28594851 11 35 theme serum 1577:1581 arg1 levels 1583:1588 the serum levels 1573:1588 the serum levels of two IgM antibodies 1573:1610 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 4 36 theme anti-carbohydrate 584:600 arg1 antibodies 602:611 anti-carbohydrate antibodies 584:611 anti-carbohydrate antibodies 584:611 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 10 37 theme high 1484:1487 arg1 sensitivity 1489:1499 significantly high sensitivity 1470:1499 significantly high sensitivity 1470:1499 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 2 38 theme tumor 331:335 arg1 progression 337:347 tumor progression 331:347 tumor progression 331:347 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 9 39 theme relative 1175:1182 arg1 abundances 1184:1193 The relative abundances 1171:1193 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation 1171:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 4 40 theme normal 618:623 arg1 populations 625:635 normal populations 618:635 normal populations 618:635 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 5 41 theme chemical 760:767 arg1 digestion 803:811 chemical reduction, alkylation and trypsin digestion 760:811 chemical reduction, alkylation and trypsin digestion 760:811 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 9 42 theme serum 1322:1326 arg1 N-glycomes 1328:1337 serum N-glycomes 1322:1337 serum N-glycomes of OSCC 1322:1345 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 12 43 theme OSCC 1850:1853 arg1 diagnosis 1855:1863 OSCC diagnosis 1850:1863 OSCC diagnosis 1850:1863 Taken together, these serum N-glycans and antibodies identified in this study should be considered as the candidates of potential biomarkers for OSCC diagnosis. 28594851 4 44 from patients 677:684 arg1 N-glycomes 569:578 serum N-glycomes 563:578 serum N-glycomes 563:578 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 4 44 from patients 677:684 arg1 antibodies 602:611 anti-carbohydrate antibodies 584:611 anti-carbohydrate antibodies 584:611 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 3 45 theme potential 505:513 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 45 theme potential 505:513 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 45 theme potential 505:513 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 46 theme glycan 424:429 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 46 theme glycan 424:429 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 46 theme glycan 424:429 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 5 47 theme reduction 769:777 arg1 digestion 803:811 chemical reduction, alkylation and trypsin digestion 760:811 chemical reduction, alkylation and trypsin digestion 760:811 Total serum proteins were lyophilized and subjected to chemical reduction, alkylation and trypsin digestion. 28594851 10 48 theme OCCC 1398:1401 arg1 patients 1403:1410 forty-eight OCCC patients 1386:1410 forty-eight OCCC patients 1386:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 3 49 theme anti-carbohydrate 472:488 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 49 theme anti-carbohydrate 472:488 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 49 theme anti-carbohydrate 472:488 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 0 50 theme anti-carbohydrate 37:53 arg1 antibody 55:62 anti-carbohydrate antibody 37:62 anti-carbohydrate antibody 37:62 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 4 51 theme serum 563:567 arg1 N-glycomes 569:578 serum N-glycomes 563:578 serum N-glycomes 563:578 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 8 52 dep % 1168:1168 arg1 75 1166:1167 75 1166:1167 75 1166:1167 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 9 53 theme total 1198:1202 arg1 glycans 1238:1244 total tri-antennary and tetra-antennary glycans 1198:1244 total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation 1198:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 8 54 dep accuracy 1144:1151 arg1 % 1168:1168 % 1168:1168 % 1168:1168 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 9 55 theme tri-antennary 1204:1216 arg1 glycans 1238:1244 total tri-antennary and tetra-antennary glycans 1198:1244 total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation 1198:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 3 56 theme cancer 530:535 arg1 diagnosis 537:545 cancer diagnosis 530:545 cancer diagnosis 530:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 57 theme aberrant 415:422 arg1 antibodies 490:499 the induced corresponding anti-carbohydrate antibodies 446:499 the induced corresponding anti-carbohydrate antibodies 446:499 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 57 theme aberrant 415:422 arg1 biomarkers 515:524 potential biomarkers 505:524 potential biomarkers for cancer diagnosis 505:545 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 3 57 theme aberrant 415:422 arg1 structures 431:440 aberrant glycan structures 415:440 aberrant glycan structures 415:440 Hence, aberrant glycan structures and the induced corresponding anti-carbohydrate antibodies are potential biomarkers for cancer diagnosis. 28594851 9 58 theme OSCC 1342:1345 arg1 N-glycomes 1328:1337 serum N-glycomes 1322:1337 serum N-glycomes of OSCC 1322:1345 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 11 59 theme patient 1690:1696 arg1 serum 1698:1702 patient serum 1690:1702 patient serum 1690:1702 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 1 60 theme protein 135:141 arg1 Glycosylation 116:128 Glycosylation 116:128 Glycosylation 116:128 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 1 60 theme protein 135:141 arg1 modification 162:173 a protein post translational modification 133:173 a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover 133:262 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 11 61 theme IgM 1597:1599 arg1 antibodies 1601:1610 two IgM antibodies 1593:1610 two IgM antibodies 1593:1610 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 11 62 theme IgG 1672:1674 arg1 antibodies 1676:1685 nine IgG antibodies 1667:1685 nine IgG antibodies 1667:1685 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 11 63 theme antibodies 1676:1685 arg1 levels 1657:1662 the decreased levels 1643:1662 the decreased levels of nine IgG antibodies in patient serum 1643:1702 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 2 64 theme glycosylation 287:299 arg1 phenomenon 313:322 a common phenomenon 304:322 a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis 304:405 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 2 64 theme glycosylation 287:299 arg1 Alteration 265:274 Alteration 265:274 Alteration of protein glycosylation 265:299 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 9 65 theme tetra-antennary 1222:1236 arg1 glycans 1238:1244 total tri-antennary and tetra-antennary glycans 1198:1244 total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation 1198:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 1 66 theme post 143:146 arg1 Glycosylation 116:128 Glycosylation 116:128 Glycosylation 116:128 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 1 66 theme post 143:146 arg1 modification 162:173 a protein post translational modification 133:173 a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover 133:262 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 9 67 theme varying 1251:1257 arg1 degrees 1259:1265 varying degrees 1251:1265 varying degrees of fucosylation and sialylation 1251:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 4 68 theme carcinoma 660:668 arg1 patients 677:684 oral squamous cell carcinoma (OSCC) patients 641:684 oral squamous cell carcinoma (OSCC) patients 641:684 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 6 69 theme MALDI-TOF-Mass 887:900 arg1 spectrometry 902:913 MALDI-TOF-Mass spectrometry 887:913 MALDI-TOF-Mass spectrometry 887:913 The N-glycans were released, purified, permethylated, and analyzed using MALDI-TOF-Mass spectrometry. 28594851 2 70 theme protein 279:285 arg1 glycosylation 287:299 protein glycosylation 279:299 protein glycosylation 279:299 Alteration of protein glycosylation is a common phenomenon during tumor progression, migration, invasion, angiogenesis, as well as metastasis. 28594851 9 71 theme glycans 1238:1244 arg1 abundances 1184:1193 The relative abundances 1171:1193 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation 1171:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 1 72 theme translational 148:160 arg1 Glycosylation 116:128 Glycosylation 116:128 Glycosylation 116:128 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 1 72 theme translational 148:160 arg1 modification 162:173 a protein post translational modification 133:173 a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover 133:262 Glycosylation is a protein post translational modification which plays important role in protein function, stabilization, trafficking, and turnover. 28594851 10 73 theme weight 1440:1445 arg1 N-glycans 1453:1461 the high-molecular weight serum N-glycans 1421:1461 the high-molecular weight serum N-glycans 1421:1461 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 4 74 theme cell 655:658 arg1 carcinoma 660:668 oral squamous cell carcinoma 641:668 oral squamous cell carcinoma (OSCC) patients 641:684 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 4 74 theme cell 655:658 arg1 OSCC 671:674 OSCC 671:674 OSCC 671:674 In this study, serum N-glycomes and anti-carbohydrate antibodies from normal populations and oral squamous cell carcinoma (OSCC) patients were investigated. 28594851 0 75 theme squamous 82:89 arg1 carcinoma 96:104 oral squamous cell carcinoma 77:104 oral squamous cell carcinoma patients 77:113 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 10 76 theme independent 1354:1364 arg1 patients 1403:1410 forty-eight OCCC patients 1386:1410 forty-eight OCCC patients 1386:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 10 76 theme independent 1354:1364 arg1 group 1377:1381 an independent validation group 1351:1381 an independent validation group of forty-eight OCCC patients 1351:1410 In an independent validation group of forty-eight OCCC patients, most of the high-molecular weight serum N-glycans showed significantly high sensitivity and specificity according to the identified cutoff values. 28594851 11 77 theme antibodies 1601:1610 arg1 levels 1583:1588 the serum levels 1573:1588 the serum levels of two IgM antibodies 1573:1610 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 28594851 9 78 with glycans 1238:1244 arg1 degrees 1259:1265 varying degrees 1251:1265 varying degrees of fucosylation and sialylation 1251:1297 The relative abundances of total tri-antennary and tetra-antennary glycans with varying degrees of fucosylation and sialylation were also increased in serum N-glycomes of OSCC. 28594851 0 79 theme oral 77:80 arg1 carcinoma 96:104 oral squamous cell carcinoma 77:104 oral squamous cell carcinoma patients 77:113 Serum N-glycome characterization and anti-carbohydrate antibody profiling in oral squamous cell carcinoma patients. 28594851 8 80 theme relative 1044:1051 arg1 abundances 1053:1062 the relative abundances 1040:1062 the relative abundances of seven N-glycans 1040:1081 We found that the relative abundances of seven N-glycans were decreased or increased in serum of OSCC with diagnostic accuracy greater than 75%. 28594851 7 81 theme antibody 957:964 arg1 profiles 966:973 the serum anti-carbohydrate antibody profiles 929:973 the serum anti-carbohydrate antibody profiles 929:973 In addition, the serum anti-carbohydrate antibody profiles were also investigated by carbohydrate microarray. 28594851 11 82 theme decreased 1647:1655 arg1 levels 1657:1662 the decreased levels 1643:1662 the decreased levels of nine IgG antibodies in patient serum 1643:1702 Furthermore, the serum levels of two IgM antibodies were elevated accompanied with the decreased levels of nine IgG antibodies in patient serum. 25978763 2 0 used used 430:433 arg2 LC-MS/MS 370:377 LC-MS/MS 370:377 LC-MS/MS with electron-transfer dissociation (ETD) 370:419 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 1 theme side 274:277 arg1 chains 279:284 Cys side chains 270:284 Cys side chains 270:284 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 1 2 theme sulfhydryls 177:187 arg1 alkylation 158:167 the alkylation 154:167 the alkylation of free sulfhydryls in proteomic experiments 154:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 2 3 theme peptides 312:319 arg1 derivatization 252:265 incomplete derivatization 241:265 incomplete derivatization of Cys side chains 241:284 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 3 theme peptides 312:319 arg1 overalkylation 290:303 overalkylation 290:303 overalkylation of the peptides 290:319 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 1 4 theme proteomic 192:200 arg1 experiments 202:212 proteomic experiments 192:212 proteomic experiments 192:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 5 5 from sulfoxide 1258:1266 arg1 loss 1244:1247 the side chain loss 1229:1247 the side chain loss from Met sulfoxide 1229:1266 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 6 theme dissociation 665:676 arg1 " 677:677 "higher-energy C-trap dissociation" 643:677 "higher-energy C-trap dissociation" (HCD) 643:683 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 6 theme dissociation 665:676 arg1 dissociation 720:731 beam-type collision-induced dissociation 692:731 beam-type collision-induced dissociation 692:731 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 6 theme dissociation 665:676 arg1 HCD 680:682 HCD 680:682 HCD 680:682 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 5 7 theme Y1-48 1089:1093 arg1 ion 1098:1100 an abundant Y1-48 Da ion 1077:1100 an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence) 1077:1194 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 8 from spectra 1032:1038 arg1 fragment 1010:1017 the normal Y1 fragment 996:1017 the normal Y1 fragment from the HCD spectra of N-linked glycopeptides 996:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 9 theme residues 854:861 arg1 carbamidomethylation 826:845 the carbamidomethylation 822:845 the carbamidomethylation of Met residues 822:861 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 10 link N-linked 1043:1050 arg1 glycopeptides 1052:1064 N-linked glycopeptides 1043:1064 N-linked glycopeptides 1043:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 6 11 theme ETD 1297:1299 arg1 data 1301:1304 good quality ETD data 1284:1304 good quality ETD data 1284:1304 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 6 12 theme good 1284:1287 arg1 data 1301:1304 good quality ETD data 1284:1304 good quality ETD data 1284:1304 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 1 13 from experiments 202:212 arg1 alkylation 158:167 the alkylation 154:167 the alkylation of free sulfhydryls in proteomic experiments 154:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 4 14 theme database 747:754 arg1 searches 756:763 the database searches 743:763 the database searches with ETD data 743:777 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 15 theme beam-type 692:700 arg1 " 677:677 "higher-energy C-trap dissociation" 643:677 "higher-energy C-trap dissociation" (HCD) 643:683 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 15 theme beam-type 692:700 arg1 dissociation 720:731 beam-type collision-induced dissociation 692:731 beam-type collision-induced dissociation 692:731 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 2 16 theme incomplete 241:250 arg1 derivatization 252:265 incomplete derivatization 241:265 incomplete derivatization of Cys side chains 241:284 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 4 17 from cases 806:810 arg1 decisive 789:796 decisive 789:796 decisive 789:796 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 1 18 from alkylation 158:167 arg1 experiments 202:212 proteomic experiments 192:212 proteomic experiments 192:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 5 19 theme fixed 917:921 arg1 charge 932:937 the resulting fixed positive charge 903:937 the resulting fixed positive charge 903:937 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 20 theme Met 1254:1256 arg1 sulfoxide 1258:1266 Met sulfoxide 1254:1266 Met sulfoxide 1254:1266 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 21 theme nominal 1115:1121 arg1 difference 1128:1137 the nominal mass difference 1111:1137 the nominal mass difference 1111:1137 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 22 theme Y1 1007:1008 arg1 fragment 1010:1017 the normal Y1 fragment 996:1017 the normal Y1 fragment from the HCD spectra of N-linked glycopeptides 996:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 2 23 theme structural 443:452 arg1 characterization 454:469 the structural characterization 439:469 the structural characterization of glycopeptides 439:486 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 0 24 theme Glycopeptide 73:84 arg1 Analysis 86:93 Glycopeptide Analysis 73:93 Glycopeptide Analysis 73:93 Carbamidomethylation Side Reactions May Lead to Glycan Misassignments in Glycopeptide Analysis. 25978763 2 25 theme electron-transfer 384:400 arg1 dissociation 402:413 electron-transfer dissociation 384:413 electron-transfer dissociation (ETD) 384:419 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 25 theme electron-transfer 384:400 arg1 ETD 416:418 ETD 416:418 ETD 416:418 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 0 26 theme Carbamidomethylation 0:19 arg1 Reactions 26:34 Carbamidomethylation Side Reactions 0:34 Carbamidomethylation Side Reactions 0:34 Carbamidomethylation Side Reactions May Lead to Glycan Misassignments in Glycopeptide Analysis. 25978763 5 27 theme acid 1181:1184 arg1 sequence 1186:1193 the unmodified amino acid sequence 1160:1193 the unmodified amino acid sequence 1160:1193 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 28 gly glycopeptides 1052:1064 arg2 glycopeptides 1052:1064 N-linked glycopeptides 1043:1064 N-linked glycopeptides 1043:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 7 29 theme unanticipated 1535:1547 arg1 modifications 1558:1570 different unanticipated covalent modifications 1525:1570 different unanticipated covalent modifications 1525:1570 Our observations also draw attention to the underreported interference of different unanticipated covalent modifications. 25978763 1 30 theme free 172:175 arg1 sulfhydryls 177:187 free sulfhydryls 172:187 free sulfhydryls in proteomic experiments 172:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 4 31 dep " 677:677 arg1 data 733:736 data 733:736 data 733:736 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 0 32 theme Glycan 48:53 arg1 Misassignments 55:68 Glycan Misassignments 48:68 Glycan Misassignments in Glycopeptide Analysis 48:93 Carbamidomethylation Side Reactions May Lead to Glycan Misassignments in Glycopeptide Analysis. 25978763 5 33 theme unmodified 1164:1173 arg1 sequence 1186:1193 the unmodified amino acid sequence 1160:1193 the unmodified amino acid sequence 1160:1193 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 7 34 theme underreported 1495:1507 arg1 interference 1509:1520 the underreported interference 1491:1520 the underreported interference of different unanticipated covalent modifications 1491:1570 Our observations also draw attention to the underreported interference of different unanticipated covalent modifications. 25978763 5 35 theme glycopeptides 1052:1064 arg1 spectra 1032:1038 the HCD spectra 1024:1038 the HCD spectra of N-linked glycopeptides 1024:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 3 36 theme Accurate 489:496 arg1 measurements 503:514 Accurate mass measurements 489:514 Accurate mass measurements 489:514 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 3 37 theme elemental 541:549 arg1 compositions 551:562 the elemental compositions 537:562 the elemental compositions of the misidentified and correct modifications 537:609 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 3 37 theme elemental 541:549 arg1 identical 615:623 identical 615:623 identical 615:623 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 2 38 with LC-MS/MS 370:377 arg1 dissociation 402:413 electron-transfer dissociation 384:413 electron-transfer dissociation (ETD) 384:419 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 38 with LC-MS/MS 370:377 arg1 ETD 416:418 ETD 416:418 ETD 416:418 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 5 39 theme ether 882:886 arg1 formation 888:896 sulfonium ether formation 872:896 sulfonium ether formation 872:896 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 40 theme HCD 1028:1030 arg1 spectra 1032:1038 the HCD spectra 1024:1038 the HCD spectra of N-linked glycopeptides 1024:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 6 41 theme such 1272:1275 arg1 cases 1277:1281 such cases 1272:1281 such cases 1272:1281 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 2 42 theme glycoforms 354:363 arg1 misassignment 337:349 the misassignment 333:349 the misassignment of glycoforms 333:363 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 6 43 theme precursor 1435:1443 arg1 ions 1445:1448 the charge-reduced precursor ions 1416:1448 the charge-reduced precursor ions 1416:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 2 44 theme chains 279:284 arg1 derivatization 252:265 incomplete derivatization 241:265 incomplete derivatization of Cys side chains 241:284 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 44 theme chains 279:284 arg1 overalkylation 290:303 overalkylation 290:303 overalkylation of the peptides 290:319 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 5 45 theme sulfonium 872:880 arg1 formation 888:896 sulfonium ether formation 872:896 sulfonium ether formation 872:896 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 3 46 theme the 567:569 arg1 compositions 551:562 the elemental compositions 537:562 the elemental compositions of the misidentified and correct modifications 537:609 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 3 46 theme the 567:569 arg1 identical 615:623 identical 615:623 identical 615:623 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 5 47 theme Met 850:852 arg1 residues 854:861 Met residues 850:861 Met residues 850:861 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 3 48 theme correct 589:595 arg1 modifications 597:609 correct modifications 589:609 correct modifications 589:609 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 4 49 theme C-trap 658:663 arg1 " 677:677 "higher-energy C-trap dissociation" 643:677 "higher-energy C-trap dissociation" (HCD) 643:683 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 49 theme C-trap 658:663 arg1 dissociation 720:731 beam-type collision-induced dissociation 692:731 beam-type collision-induced dissociation 692:731 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 49 theme C-trap 658:663 arg1 HCD 680:682 HCD 680:682 HCD 680:682 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 5 50 theme Da 1095:1096 arg1 ion 1098:1100 an abundant Y1-48 Da ion 1077:1100 an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence) 1077:1194 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 51 theme " 677:677 arg1 Incorporation 626:638 Incorporation 626:638 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data 626:777 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 6 52 theme quality 1289:1295 arg1 data 1301:1304 good quality ETD data 1284:1304 good quality ETD data 1284:1304 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 3 53 theme modifications 597:609 arg1 compositions 551:562 the elemental compositions 537:562 the elemental compositions of the misidentified and correct modifications 537:609 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 3 53 theme modifications 597:609 arg1 identical 615:623 identical 615:623 identical 615:623 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 5 54 theme positive 923:930 arg1 charge 932:937 the resulting fixed positive charge 903:937 the resulting fixed positive charge 903:937 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 55 with searches 756:763 arg1 data 774:777 ETD data 770:777 ETD data 770:777 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 2 56 theme Cys 270:272 arg1 chains 279:284 Cys side chains 270:284 Cys side chains 270:284 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 1 57 theme used 137:140 arg1 reagent 142:148 the most widely used reagent 121:148 the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments 121:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 1 57 theme used 137:140 arg1 Iodoacetamide 96:108 Iodoacetamide 96:108 Iodoacetamide 96:108 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 6 58 theme abundant 1358:1365 arg1 fragments 1367:1375 abundant fragments 1358:1375 abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions 1358:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 5 59 theme abundant 1080:1087 arg1 ion 1098:1100 an abundant Y1-48 Da ion 1077:1100 an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence) 1077:1194 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 60 theme chain 1238:1242 arg1 loss 1244:1247 the side chain loss 1229:1247 the side chain loss from Met sulfoxide 1229:1266 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 1 61 from sulfhydryls 177:187 arg1 experiments 202:212 proteomic experiments 192:212 proteomic experiments 192:212 Iodoacetamide is perhaps the most widely used reagent for the alkylation of free sulfhydryls in proteomic experiments. 25978763 5 62 theme resulting 907:915 arg1 charge 932:937 the resulting fixed positive charge 903:937 the resulting fixed positive charge 903:937 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 63 theme collision-induced 702:718 arg1 " 677:677 "higher-energy C-trap dissociation" 643:677 "higher-energy C-trap dissociation" (HCD) 643:683 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 63 theme collision-induced 702:718 arg1 dissociation 720:731 beam-type collision-induced dissociation 692:731 beam-type collision-induced dissociation 692:731 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 64 from decisive 789:796 arg1 cases 806:810 most cases 801:810 most cases 801:810 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 7 65 theme modifications 1558:1570 arg1 interference 1509:1520 the underreported interference 1491:1520 the underreported interference of different unanticipated covalent modifications 1491:1570 Our observations also draw attention to the underreported interference of different unanticipated covalent modifications. 25978763 0 66 from Misassignments 55:68 arg1 Analysis 86:93 Glycopeptide Analysis 73:93 Glycopeptide Analysis 73:93 Carbamidomethylation Side Reactions May Lead to Glycan Misassignments in Glycopeptide Analysis. 25978763 5 67 theme normal 1000:1005 arg1 fragment 1010:1017 the normal Y1 fragment 996:1017 the normal Y1 fragment from the HCD spectra of N-linked glycopeptides 996:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 0 68 theme Side 21:24 arg1 Reactions 26:34 Carbamidomethylation Side Reactions 0:34 Carbamidomethylation Side Reactions 0:34 Carbamidomethylation Side Reactions May Lead to Glycan Misassignments in Glycopeptide Analysis. 25978763 5 69 theme mass 1123:1126 arg1 difference 1128:1137 the nominal mass difference 1111:1137 the nominal mass difference 1111:1137 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 70 theme characteristic 950:963 arg1 fragmentation 965:977 a characteristic fragmentation 948:977 a characteristic fragmentation 948:977 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 71 theme ETD 770:772 arg1 data 774:777 ETD data 770:777 ETD data 770:777 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 6 72 theme CH3-S-CH2CONH2 1384:1397 arg1 elimination 1399:1409 CH3-S-CH2CONH2 elimination 1384:1409 CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions 1384:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 7 73 theme covalent 1549:1556 arg1 modifications 1558:1570 different unanticipated covalent modifications 1525:1570 different unanticipated covalent modifications 1525:1570 Our observations also draw attention to the underreported interference of different unanticipated covalent modifications. 25978763 2 74 gly glycopeptides 474:486 arg2 glycopeptides 474:486 glycopeptides 474:486 glycopeptides 474:486 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 2 75 theme glycopeptides 474:486 arg1 characterization 454:469 the structural characterization 439:469 the structural characterization of glycopeptides 439:486 Here, we report that both incomplete derivatization of Cys side chains and overalkylation of the peptides may lead to the misassignment of glycoforms when LC-MS/MS with electron-transfer dissociation (ETD) alone is used for the structural characterization of glycopeptides. 25978763 6 76 theme due 1377:1379 arg1 fragments 1367:1375 abundant fragments 1358:1375 abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions 1358:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 7 77 theme different 1525:1533 arg1 modifications 1558:1570 different unanticipated covalent modifications 1525:1570 different unanticipated covalent modifications 1525:1570 Our observations also draw attention to the underreported interference of different unanticipated covalent modifications. 25978763 5 78 theme N-linked 1043:1050 arg1 glycopeptides 1052:1064 N-linked glycopeptides 1043:1064 N-linked glycopeptides 1043:1064 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 5 79 theme amino 1175:1179 arg1 sequence 1186:1193 the unmodified amino acid sequence 1160:1193 the unmodified amino acid sequence 1160:1193 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 6 80 from ions 1445:1448 arg1 elimination 1399:1409 CH3-S-CH2CONH2 elimination 1384:1409 CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions 1384:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 5 81 dep ion 1098:1100 arg1 given 1142:1146 given 1142:1146 is given relative to the unmodified amino acid sequence 1139:1193 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 25978763 4 82 theme most 801:804 arg1 cases 806:810 most cases 801:810 most cases 801:810 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 3 83 theme mass 498:501 arg1 measurements 503:514 Accurate mass measurements 489:514 Accurate mass measurements 489:514 Accurate mass measurements do not help, because the elemental compositions of the misidentified and correct modifications are identical. 25978763 4 84 theme higher-energy 644:656 arg1 " 677:677 "higher-energy C-trap dissociation" 643:677 "higher-energy C-trap dissociation" (HCD) 643:683 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 84 theme higher-energy 644:656 arg1 dissociation 720:731 beam-type collision-induced dissociation 692:731 beam-type collision-induced dissociation 692:731 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 4 84 theme higher-energy 644:656 arg1 HCD 680:682 HCD 680:682 HCD 680:682 Incorporation of "higher-energy C-trap dissociation" (HCD), i.e., beam-type collision-induced dissociation data into the database searches with ETD data may prove decisive in most cases. 25978763 6 85 theme charge-reduced 1420:1433 arg1 ions 1445:1448 the charge-reduced precursor ions 1416:1448 the charge-reduced precursor ions 1416:1448 In such cases, good quality ETD data may indicate the discrepancy, and will also display abundant fragments due to CH3-S-CH2CONH2 elimination from the charge-reduced precursor ions. 25978763 5 86 theme side 1233:1236 arg1 chain 1238:1242 the side chain 1229:1242 the side chain loss from Met sulfoxide 1229:1266 However, the carbamidomethylation of Met residues leads to sulfonium ether formation, and the resulting fixed positive charge triggers a characteristic fragmentation, that eliminates the normal Y1 fragment from the HCD spectra of N-linked glycopeptides, producing an abundant Y1-48 Da ion instead (the nominal mass difference is given relative to the unmodified amino acid sequence), that easily can be mistaken for the side chain loss from Met sulfoxide. 28109443 5 0 theme IgA1 1284:1287 arg1 fractions 1289:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 1 theme systematic 1051:1060 arg1 study 1062:1066 a systematic study 1049:1066 a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls 1049:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 2 2 theme SialylTn 585:592 arg1 antigen 600:606 the SialylTn (STn) antigen 581:606 its sialylated version the SialylTn (STn) antigen 558:606 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 3 dep version 573:579 arg1 antigen 600:606 the SialylTn (STn) antigen 581:606 its sialylated version the SialylTn (STn) antigen 558:606 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 4 theme homogeneous 1642:1652 arg1 antigens 1661:1668 homogeneous Tn/STn antigens 1642:1668 homogeneous Tn/STn antigens 1642:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 5 from patients 1115:1122 arg1 IgA1 1095:1098 plasma IgA1 1088:1098 plasma IgA1 from both IgAN patients and healthy controls 1088:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 5 from patients 1115:1122 arg1 O-glycans 1075:1083 the O-glycans 1071:1083 the O-glycans of plasma IgA1 from both IgAN patients and healthy controls 1071:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 6 6 theme minor 1360:1364 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 7 contain had 1428:1430 arg2 antigens 1439:1446 Tn/STn antigens 1432:1446 Tn/STn antigens 1432:1446 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 7 contain had 1428:1430 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 2 8 theme truncated 511:519 arg1 O-glycans 521:529 undergalactosylated or truncated O-glycans 488:529 undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR 488:616 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 0 9 theme O-Glycan 116:123 arg1 Analysis 125:132 Mass Spectrometry O-Glycan Analysis 98:132 Mass Spectrometry O-Glycan Analysis 98:132 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 2 10 theme undergalactosylated 488:506 arg1 O-glycans 521:529 undergalactosylated or truncated O-glycans 488:529 undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR 488:616 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 11 theme distinct 1523:1530 arg1 glycoform 1627:1635 one minor glycoform 1617:1635 one minor glycoform with homogeneous Tn/STn antigens 1617:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 11 theme distinct 1523:1530 arg1 glycoform 1556:1564 one major glycoform 1546:1564 one major glycoform with homogeneous normal core 1-based O-glycans 1546:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 11 theme distinct 1523:1530 arg1 O-glycoforms 1532:1543 only two distinct O-glycoforms 1514:1543 only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens 1514:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 3 12 with patients 710:717 arg1 IgAN 724:727 IgAN 724:727 IgAN 724:727 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 1 13 theme mucin-type 193:202 arg1 O-glycans 215:223 O-glycans 215:223 O-glycans 215:223 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 13 theme mucin-type 193:202 arg1 O-glycans 204:212 four to six mucin-type O-glycans 181:212 four to six mucin-type O-glycans (O-glycans) on its hinge region (HR) 181:249 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 9 14 gly O-glycosylated 2021:2034 arg1 proteins 2036:2043 any other O-glycosylated proteins 2011:2043 any other O-glycosylated proteins 2011:2043 The method could also be applied to the analysis of any other O-glycosylated proteins. 28109443 1 15 from O-glycans 204:212 arg1 HR 247:248 HR 247:248 HR 247:248 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 15 from O-glycans 204:212 arg1 region 239:244 its hinge region 229:244 its hinge region (HR) 229:249 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 5 16 from O-glycans 1075:1083 arg1 patients 1115:1122 IgAN patients 1110:1122 IgAN patients 1110:1122 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 16 from O-glycans 1075:1083 arg1 controls 1136:1143 healthy controls 1128:1143 healthy controls 1128:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 17 theme IgAN 1110:1113 arg1 patients 1115:1122 IgAN patients 1110:1122 IgAN patients 1110:1122 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 1 18 theme Human 135:139 arg1 A1 156:157 Human immunoglobulin A1 135:157 Human immunoglobulin A1 (IgA1) 135:164 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 18 theme Human 135:139 arg1 O-glycoprotein 273:286 the most abundant O-glycoprotein 255:286 the most abundant O-glycoprotein in plasma or serum 255:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 18 theme Human 135:139 arg1 IgA1 160:163 IgA1 160:163 IgA1 160:163 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 2 19 theme IgA 457:459 arg1 IgAN 474:477 IgAN 474:477 IgAN 474:477 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 19 theme IgA 457:459 arg1 nephropathy 461:471 IgA nephropathy 457:471 IgA nephropathy (IgAN) 457:478 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 9 20 theme other 2015:2019 arg1 proteins 2036:2043 any other O-glycosylated proteins 2011:2043 any other O-glycosylated proteins 2011:2043 The method could also be applied to the analysis of any other O-glycosylated proteins. 28109443 8 21 dep methodology 1918:1928 arg1 developed 1933:1941 developed 1933:1941 developed in more detail 1933:1956 Herein, we describe the methodology we developed in more detail. 28109443 6 22 theme IgAN 1393:1396 arg1 patients 1398:1405 IgAN patients 1393:1405 IgAN patients 1393:1405 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 7 23 from role 1721:1724 arg1 pathogenesis 1756:1767 pathogenesis 1756:1767 pathogenesis of IgAN 1756:1775 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 7 23 from role 1721:1724 arg1 IgA1 1748:1751 IgA1 1748:1751 IgA1 1748:1751 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 6 24 theme normal 1583:1588 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 25 with glycoform 1556:1564 arg1 antigens 1661:1668 homogeneous Tn/STn antigens 1642:1668 homogeneous Tn/STn antigens 1642:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 25 with glycoform 1556:1564 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 26 theme plasma 1088:1093 arg1 IgA1 1095:1098 plasma IgA1 1088:1098 plasma IgA1 from both IgAN patients and healthy controls 1088:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 9 27 theme proteins 2036:2043 arg1 analysis 1999:2006 the analysis 1995:2006 the analysis of any other O-glycosylated proteins 1995:2043 The method could also be applied to the analysis of any other O-glycosylated proteins. 28109443 0 28 theme Lectin 64:69 arg1 Chromatography 71:84 Serial Lectin Chromatography 57:84 Serial Lectin Chromatography 57:84 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 2 29 with patients 443:450 arg1 IgAN 474:477 IgAN 474:477 IgAN 474:477 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 29 with patients 443:450 arg1 nephropathy 461:471 IgA nephropathy 457:471 IgA nephropathy (IgAN) 457:478 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 30 from patients 1398:1405 arg1 IgA1 1378:1381 IgA1 1378:1381 IgA1 from both IgAN patients and healthy controls 1378:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 30 from patients 1398:1405 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 31 with glycoform 1627:1635 arg1 antigens 1661:1668 homogeneous Tn/STn antigens 1642:1668 homogeneous Tn/STn antigens 1642:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 31 with glycoform 1627:1635 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 32 theme O-glycoprotein 1462:1475 arg1 molecules 1482:1490 the O-glycoprotein IgA1 molecules 1458:1490 the O-glycoprotein IgA1 molecules from most samples 1458:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 33 from fraction 1366:1373 arg1 patients 1398:1405 IgAN patients 1393:1405 IgAN patients 1393:1405 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 33 from fraction 1366:1373 arg1 controls 1419:1426 healthy controls 1411:1426 healthy controls 1411:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 3 34 theme healthy 787:793 arg1 individuals 795:805 healthy individuals 787:805 healthy individuals 787:805 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 6 35 theme major 1550:1554 arg1 glycoform 1556:1564 one major glycoform 1546:1564 one major glycoform with homogeneous normal core 1-based O-glycans 1546:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 35 theme major 1550:1554 arg1 O-glycoforms 1532:1543 only two distinct O-glycoforms 1514:1543 only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens 1514:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 3 36 from Tn/STn 656:661 arg1 HR 670:671 the HR 666:671 the HR of IgA1 666:679 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 0 37 theme Mass 98:101 arg1 Analysis 125:132 Mass Spectrometry O-Glycan Analysis 98:132 Mass Spectrometry O-Glycan Analysis 98:132 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 4 38 theme IgA1 891:894 arg1 molecule 896:903 one IgA1 molecule 887:903 one IgA1 molecule 887:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 6 39 theme minor 1621:1625 arg1 glycoform 1627:1635 one minor glycoform 1617:1635 one minor glycoform with homogeneous Tn/STn antigens 1617:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 39 theme minor 1621:1625 arg1 O-glycoforms 1532:1543 only two distinct O-glycoforms 1514:1543 only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens 1514:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 40 theme lectin 1170:1175 arg1 chromatography 1177:1190 serial HPA and PNA lectin chromatography 1151:1190 chromatography 1177:1190 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 3 41 from patients 710:717 arg1 IgA1 700:703 the IgA1 696:703 the IgA1 from patients with IgAN 696:727 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 2 42 theme complex 384:390 arg1 structures 392:401 core 1-based complex structures 371:401 core 1-based complex structures 371:401 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 42 theme complex 384:390 arg1 O-glycans 321:329 normal O-glycans 314:329 normal O-glycans from hematopoietic-originated cells 314:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 43 theme IgA1 1378:1381 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 44 theme serial 1151:1156 arg1 HPA 1158:1160 serial HPA and PNA lectin chromatography 1151:1190 HPA 1158:1160 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 2 45 theme core 371:374 arg1 structures 392:401 core 1-based complex structures 371:401 core 1-based complex structures 371:401 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 45 theme core 371:374 arg1 O-glycans 321:329 normal O-glycans 314:329 normal O-glycans from hematopoietic-originated cells 314:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 0 46 theme O-Glycoforms 27:38 arg1 Separation 0:9 Separation 0:9 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography 0:84 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 7 47 theme Tn/STn 1729:1734 arg1 antigens 1736:1743 Tn/STn antigens 1729:1743 Tn/STn antigens on IgA1 1729:1751 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 0 48 theme Human 43:47 arg1 IgA1 49:52 Human IgA1 43:52 Human IgA1 43:52 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 3 49 dep debate 641:646 arg1 specific 684:691 specific 684:691 specific 684:691 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 2 50 from cells 361:365 arg1 O-glycans 321:329 normal O-glycans 314:329 normal O-glycans from hematopoietic-originated cells 314:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 50 from cells 361:365 arg1 structures 392:401 core 1-based complex structures 371:401 core 1-based complex structures 371:401 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 51 theme 1-based 1595:1601 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 1 52 dep six 189:191 arg1 to 186:187 to 186:187 to 186:187 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 2 53 theme normal 314:319 arg1 O-glycans 321:329 normal O-glycans 314:329 normal O-glycans from hematopoietic-originated cells 314:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 53 theme normal 314:319 arg1 structures 392:401 core 1-based complex structures 371:401 core 1-based complex structures 371:401 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 1 54 from O-glycoprotein 273:286 arg1 plasma 291:296 plasma 291:296 plasma 291:296 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 54 from O-glycoprotein 273:286 arg1 serum 301:305 serum 301:305 serum 301:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 8 55 theme more 1946:1949 arg1 detail 1951:1956 more detail 1946:1956 more detail 1946:1956 Herein, we describe the methodology we developed in more detail. 28109443 5 56 theme western 1204:1210 arg1 blotting 1212:1219 western blotting 1204:1219 western blotting 1204:1219 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 4 57 from O-glycans 846:854 arg1 HRs 880:882 the two HRs 872:882 the two HRs of one IgA1 molecule 872:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 6 58 contain had 1510:1512 arg2 O-glycoforms 1532:1543 only two distinct O-glycoforms 1514:1543 only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens 1514:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 58 contain had 1510:1512 arg2 glycoform 1627:1635 one minor glycoform 1617:1635 one minor glycoform with homogeneous Tn/STn antigens 1617:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 58 contain had 1510:1512 arg2 glycoform 1556:1564 one major glycoform 1546:1564 one major glycoform with homogeneous normal core 1-based O-glycans 1546:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 58 contain had 1510:1512 arg1 molecules 1482:1490 the O-glycoprotein IgA1 molecules 1458:1490 the O-glycoprotein IgA1 molecules from most samples 1458:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 59 theme PNA-bound 1274:1282 arg1 fractions 1289:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 1 60 theme hinge 233:237 arg1 HR 247:248 HR 247:248 HR 247:248 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 60 theme hinge 233:237 arg1 region 239:244 its hinge region 229:244 its hinge region (HR) 229:249 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 2 61 theme Tn 543:544 arg1 antigen 546:552 the Tn antigen 539:552 the Tn antigen 539:552 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 6 62 theme Tn/STn 1654:1659 arg1 antigens 1661:1668 homogeneous Tn/STn antigens 1642:1668 homogeneous Tn/STn antigens 1642:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 1 63 gly O-glycoprotein 273:286 arg1 A1 156:157 Human immunoglobulin A1 135:157 Human immunoglobulin A1 (IgA1) 135:164 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 63 gly O-glycoprotein 273:286 arg1 O-glycoprotein 273:286 the most abundant O-glycoprotein 255:286 the most abundant O-glycoprotein in plasma or serum 255:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 5 64 theme O-glycans 1245:1253 arg1 blotting 1212:1219 western blotting 1204:1219 western blotting 1204:1219 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 64 theme O-glycans 1245:1253 arg1 analysis 1233:1240 further analysis 1225:1240 further analysis of O-glycans 1225:1253 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 4 65 dep O-glycans 996:1004 arg1 both 973:976 both 973:976 both 973:976 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 65 dep O-glycans 996:1004 arg1 Tn/STn 989:994 Tn/STn 989:994 Tn/STn 989:994 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 2 66 gly sialylated 562:571 arg1 version 573:579 its sialylated version 558:579 its sialylated version the SialylTn (STn) antigen 558:606 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 7 67 from IgA1 1748:1751 arg1 role 1721:1724 the role 1717:1724 the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN 1717:1775 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 2 68 theme sialylated 562:571 arg1 version 573:579 its sialylated version 558:579 its sialylated version the SialylTn (STn) antigen 558:606 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 5 69 theme HPA-bound 1260:1268 arg1 fractions 1289:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 HPA-bound and PNA-bound IgA1 fractions 1260:1297 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 6 70 theme variable 1351:1358 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 7 71 theme practical 1806:1814 arg1 methodology 1816:1826 a practical methodology 1804:1826 a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1 1804:1891 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 2 72 from patients 443:450 arg1 IgA1 433:436 the IgA1 429:436 the IgA1 from patients with IgA nephropathy (IgAN) 429:478 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 4 73 theme additional 811:820 arg1 question 822:829 An additional question 808:829 An additional question 808:829 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 1 74 theme abundant 264:271 arg1 A1 156:157 Human immunoglobulin A1 135:157 Human immunoglobulin A1 (IgA1) 135:164 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 74 theme abundant 264:271 arg1 O-glycoprotein 273:286 the most abundant O-glycoprotein 255:286 the most abundant O-glycoprotein in plasma or serum 255:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 4 75 dep heterogeneous 958:970 arg1 O-glycans 996:1004 both normal and Tn/STn O-glycans 973:1004 both normal and Tn/STn O-glycans 973:1004 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 6 76 from controls 1419:1426 arg1 IgA1 1378:1381 IgA1 1378:1381 IgA1 from both IgAN patients and healthy controls 1378:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 76 from controls 1419:1426 arg1 fraction 1366:1373 a variable minor fraction 1349:1373 a variable minor fraction of IgA1 from both IgAN patients and healthy controls 1349:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 77 dep O-glycoforms 1532:1543 arg1 glycoform 1627:1635 one minor glycoform 1617:1635 one minor glycoform with homogeneous Tn/STn antigens 1617:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 77 dep O-glycoforms 1532:1543 arg1 glycoform 1556:1564 one major glycoform 1546:1564 one major glycoform with homogeneous normal core 1-based O-glycans 1546:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 77 dep O-glycoforms 1532:1543 arg1 O-glycoforms 1532:1543 only two distinct O-glycoforms 1514:1543 only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens 1514:1668 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 78 from controls 1136:1143 arg1 IgA1 1095:1098 plasma IgA1 1088:1098 plasma IgA1 from both IgAN patients and healthy controls 1088:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 78 from controls 1136:1143 arg1 O-glycans 1075:1083 the O-glycans 1071:1083 the O-glycans of plasma IgA1 from both IgAN patients and healthy controls 1071:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 3 79 theme IgA1 676:679 arg1 HR 670:671 the HR 666:671 the HR of IgA1 666:679 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 5 80 from study 1062:1066 arg1 O-glycans 1075:1083 the O-glycans 1071:1083 the O-glycans of plasma IgA1 from both IgAN patients and healthy controls 1071:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 6 81 theme healthy 1411:1417 arg1 controls 1419:1426 healthy controls 1411:1426 healthy controls 1411:1426 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 82 theme most 1497:1500 arg1 samples 1502:1508 most samples 1497:1508 most samples 1497:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 0 83 theme Serial 57:62 arg1 Chromatography 71:84 Serial Lectin Chromatography 57:84 Serial Lectin Chromatography 57:84 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 1 84 theme immunoglobulin 141:154 arg1 A1 156:157 Human immunoglobulin A1 135:157 Human immunoglobulin A1 (IgA1) 135:164 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 84 theme immunoglobulin 141:154 arg1 O-glycoprotein 273:286 the most abundant O-glycoprotein 255:286 the most abundant O-glycoprotein in plasma or serum 255:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 84 theme immunoglobulin 141:154 arg1 IgA1 160:163 IgA1 160:163 IgA1 160:163 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 6 85 theme core 1590:1593 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 9 86 theme O-glycosylated 2021:2034 arg1 proteins 2036:2043 any other O-glycosylated proteins 2011:2043 any other O-glycosylated proteins 2011:2043 The method could also be applied to the analysis of any other O-glycosylated proteins. 28109443 4 87 dep both 973:976 arg1 normal 978:983 normal 978:983 normal 978:983 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 3 88 theme serum 773:777 arg1 IgA1 779:782 serum IgA1 773:782 serum IgA1 of healthy individuals 773:805 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 6 89 theme homogeneous 1571:1581 arg1 O-glycans 1603:1611 homogeneous normal core 1-based O-glycans 1571:1611 homogeneous normal core 1-based O-glycans 1571:1611 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 7 90 theme IgA1 1888:1891 arg1 O-glycans 1875:1883 the O-glycans 1871:1883 the O-glycans of IgA1 1871:1891 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 5 91 theme IgA1 1095:1098 arg1 O-glycans 1075:1083 the O-glycans 1071:1083 the O-glycans of plasma IgA1 from both IgAN patients and healthy controls 1071:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 7 92 theme IgAN 1772:1775 arg1 pathogenesis 1756:1767 pathogenesis 1756:1767 pathogenesis of IgAN 1756:1775 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 6 93 theme IgA1 1477:1480 arg1 molecules 1482:1490 the O-glycoprotein IgA1 molecules 1458:1490 the O-glycoprotein IgA1 molecules from most samples 1458:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 7 94 from antigens 1736:1743 arg1 IgA1 1748:1751 IgA1 1748:1751 IgA1 1748:1751 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 2 95 contain carries 480:486 arg2 O-glycans 521:529 undergalactosylated or truncated O-glycans 488:529 undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR 488:616 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 95 contain carries 480:486 arg1 IgA1 433:436 the IgA1 429:436 the IgA1 from patients with IgA nephropathy (IgAN) 429:478 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 0 96 theme Spectrometry 103:114 arg1 Analysis 125:132 Mass Spectrometry O-Glycan Analysis 98:132 Mass Spectrometry O-Glycan Analysis 98:132 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 4 97 dep homogeneous 909:919 arg1 homogeneous 909:919 homogeneous 909:919 homogeneous 909:919 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 97 dep homogeneous 909:919 arg1 O-glycans 846:854 the O-glycans 842:854 the O-glycans at all sites on the two HRs of one IgA1 molecule 842:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 97 dep homogeneous 909:919 arg1 normal 933:938 normal 933:938 normal 933:938 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 97 dep homogeneous 909:919 arg1 Tn/STn 947:952 all Tn/STn 943:952 all Tn/STn 943:952 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 3 98 theme individuals 795:805 arg1 IgA1 779:782 serum IgA1 773:782 serum IgA1 of healthy individuals 773:805 Yet, there is still a debate whether Tn/STn on the HR of IgA1 is specific to the IgA1 from patients with IgAN since these antigens have also been seen in serum IgA1 of healthy individuals. 28109443 5 99 theme mass 1302:1305 arg1 spectrometry 1307:1318 mass spectrometry 1302:1318 mass spectrometry 1302:1318 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 5 100 theme PNA 1166:1168 arg1 chromatography 1177:1190 serial HPA and PNA lectin chromatography 1151:1190 chromatography 1177:1190 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 2 101 theme hematopoietic-originated 336:359 arg1 cells 361:365 hematopoietic-originated cells 336:365 hematopoietic-originated cells 336:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 102 theme many 404:407 arg1 reports 409:415 many reports 404:415 many reports 404:415 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 103 from antigen 546:552 arg1 HR 615:616 the HR 611:616 the HR 611:616 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 5 104 theme healthy 1128:1134 arg1 controls 1136:1143 healthy controls 1128:1143 healthy controls 1128:1143 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 0 105 theme Distinct 18:25 arg1 O-Glycoforms 27:38 Two Distinct O-Glycoforms 14:38 Two Distinct O-Glycoforms of Human IgA1 14:52 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 7 106 theme antigens 1736:1743 arg1 role 1721:1724 the role 1717:1724 the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN 1717:1775 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 4 107 dep normal 933:938 arg1 either 922:927 either 922:927 either 922:927 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 108 theme molecule 896:903 arg1 HRs 880:882 the two HRs 872:882 the two HRs of one IgA1 molecule 872:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 1 109 contain carries 173:179 arg2 O-glycans 215:223 O-glycans 215:223 O-glycans 215:223 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 109 contain carries 173:179 arg1 A1 156:157 Human immunoglobulin A1 135:157 Human immunoglobulin A1 (IgA1) 135:164 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 109 contain carries 173:179 arg1 O-glycoprotein 273:286 the most abundant O-glycoprotein 255:286 the most abundant O-glycoprotein in plasma or serum 255:305 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 109 contain carries 173:179 arg1 IgA1 160:163 IgA1 160:163 IgA1 160:163 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 1 109 contain carries 173:179 arg2 O-glycans 204:212 four to six mucin-type O-glycans 181:212 four to six mucin-type O-glycans (O-glycans) on its hinge region (HR) 181:249 Human immunoglobulin A1 (IgA1), which carries four to six mucin-type O-glycans (O-glycans) on its hinge region (HR), is the most abundant O-glycoprotein in plasma or serum. 28109443 2 110 theme 1-based 376:382 arg1 structures 392:401 core 1-based complex structures 371:401 core 1-based complex structures 371:401 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 110 theme 1-based 376:382 arg1 O-glycans 321:329 normal O-glycans 314:329 normal O-glycans from hematopoietic-originated cells 314:365 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 2 111 from version 573:579 arg1 HR 615:616 the HR 611:616 the HR 611:616 While normal O-glycans from hematopoietic-originated cells are core 1-based complex structures, many reports showed that the IgA1 from patients with IgA nephropathy (IgAN) carries undergalactosylated or truncated O-glycans such as the Tn antigen and its sialylated version the SialylTn (STn) antigen on the HR. 28109443 0 112 theme IgA1 49:52 arg1 O-Glycoforms 27:38 Two Distinct O-Glycoforms 14:38 Two Distinct O-Glycoforms of Human IgA1 14:52 Separation of Two Distinct O-Glycoforms of Human IgA1 by Serial Lectin Chromatography Followed by Mass Spectrometry O-Glycan Analysis. 28109443 7 113 theme serious 1694:1700 arg1 question 1702:1709 a serious question 1692:1709 a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN 1692:1775 These results raised a serious question about the role of Tn/STn antigens on IgA1 in pathogenesis of IgAN, and there is a demand for a practical methodology that any laboratory can utilize to analyze the O-glycans of IgA1. 28109443 6 114 from samples 1502:1508 arg1 molecules 1482:1490 the O-glycoprotein IgA1 molecules 1458:1490 the O-glycoprotein IgA1 molecules from most samples 1458:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 5 115 theme further 1225:1231 arg1 analysis 1233:1240 further analysis 1225:1240 further analysis of O-glycans 1225:1253 To address these questions, we conducted a systematic study on the O-glycans of plasma IgA1 from both IgAN patients and healthy controls using serial HPA and PNA lectin chromatography followed by western blotting and further analysis of O-glycans from HPA-bound and PNA-bound IgA1 fractions by mass spectrometry. 28109443 6 116 gly O-glycoprotein 1462:1475 arg1 O-glycoprotein 1462:1475 the O-glycoprotein IgA1 molecules 1458:1490 the O-glycoprotein IgA1 molecules from most samples 1458:1508 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 6 117 theme Tn/STn 1432:1437 arg1 antigens 1439:1446 Tn/STn antigens 1432:1446 Tn/STn antigens 1432:1446 Unexpectedly, we found that a variable minor fraction of IgA1 from both IgAN patients and healthy controls had Tn/STn antigens, and that the O-glycoprotein IgA1 molecules from most samples had only two distinct O-glycoforms: one major glycoform with homogeneous normal core 1-based O-glycans and one minor glycoform with homogeneous Tn/STn antigens. 28109443 4 118 from sites 863:867 arg1 homogeneous 909:919 homogeneous 909:919 homogeneous 909:919 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 118 from sites 863:867 arg1 O-glycans 846:854 the O-glycans 842:854 the O-glycans at all sites on the two HRs of one IgA1 molecule 842:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 118 from sites 863:867 arg1 normal 933:938 normal 933:938 normal 933:938 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 118 from sites 863:867 arg1 Tn/STn 947:952 all Tn/STn 943:952 all Tn/STn 943:952 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 119 theme all 943:945 arg1 homogeneous 909:919 homogeneous 909:919 homogeneous 909:919 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 119 theme all 943:945 arg1 O-glycans 846:854 the O-glycans 842:854 the O-glycans at all sites on the two HRs of one IgA1 molecule 842:903 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 119 theme all 943:945 arg1 normal 933:938 normal 933:938 normal 933:938 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 28109443 4 119 theme all 943:945 arg1 Tn/STn 947:952 all Tn/STn 943:952 all Tn/STn 943:952 An additional question is whether the O-glycans at all sites on the two HRs of one IgA1 molecule are homogeneous (either all normal or all Tn/STn) or heterogeneous (both normal and Tn/STn O-glycans). 25252784 2 0 theme GlcN 516:519 arg1 backbone 521:528 the flexible βGlcN(1→6)GlcN backbone 493:528 the flexible βGlcN(1→6)GlcN backbone of Lipid A 493:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 7 1 theme molecular 1455:1463 arg1 shape 1465:1469 the molecular shape 1451:1469 the molecular shape of its carbohydrate backbone 1451:1498 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 2 2 theme Lipid 533:537 arg1 A 539:539 Lipid A 533:539 Lipid A 533:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 3 theme αGlcN 1163:1167 arg1 A 1190:1190 αGlcN(1↔1)αMan-based Lipid A 1163:1190 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) 1163:1212 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 5 4 with acylation 987:995 arg1 acids 1026:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 2 5 theme rigid 560:564 arg1 scaffold 596:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold 558:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 558:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 1 6 theme lipopolysaccharide 136:153 arg1 A 188:188 a glycophospholipid Lipid A 162:188 a glycophospholipid Lipid A 162:188 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 1 6 theme lipopolysaccharide 136:153 arg1 portion 125:131 The endotoxic portion 111:131 The endotoxic portion of lipopolysaccharide (LPS) 111:159 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 7 7 theme A 1439:1439 arg1 flexibility 1418:1428 the conformational flexibility 1399:1428 the conformational flexibility of Lipid A 1399:1439 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 5 8 theme A 1151:1151 arg1 mimetics 1153:1160 monophosphorylated hexaacylated Lipid A mimetics 1113:1160 monophosphorylated hexaacylated Lipid A mimetics 1113:1160 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 2 9 theme A 669:669 arg1 shape 630:634 the molecular shape 616:634 the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 616:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 4 10 theme 1↔1 822:824 arg1 linkage 827:833 The α,α-(1↔1) linkage 813:833 linkage 827:833 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 0 11 theme Toll-like 80:88 arg1 receptor 90:97 potent Toll-like receptor 4 73:99 potent Toll-like receptor 4 agonists 73:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 5 12 theme -3-acyloxyacyl 1005:1018 arg1 acids 1026:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 7 13 theme efficient 1590:1598 arg1 approach 1600:1607 an efficient approach 1587:1607 an efficient approach toward powerful and adjustable TLR4 activation 1587:1654 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 2 14 theme flexible 497:504 arg1 backbone 521:528 the flexible βGlcN(1→6)GlcN backbone 493:528 the flexible βGlcN(1→6)GlcN backbone of Lipid A 493:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 2 15 theme X-ray 688:692 arg1 structure 694:702 the X-ray structure 684:702 the X-ray structure 684:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 16 theme HEK293 1291:1296 arg1 cells 1298:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 1 17 theme pro-inflammatory 317:332 arg1 signaling 341:349 pro-inflammatory immune signaling 317:349 pro-inflammatory immune signaling 317:349 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 1 18 theme glycophospholipid 164:180 arg1 A 188:188 a glycophospholipid Lipid A 162:188 a glycophospholipid Lipid A 162:188 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 1 18 theme glycophospholipid 164:180 arg1 portion 125:131 The endotoxic portion 111:131 The endotoxic portion of lipopolysaccharide (LPS) 111:159 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 4 19 theme α 817:817 arg1 linkage 827:833 The α,α-(1↔1) linkage 813:833 linkage 827:833 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 6 20 theme A 1190:1190 arg1 mimetics 1192:1199 αGlcN(1↔1)αMan-based Lipid A mimetics 1163:1199 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) 1163:1212 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 3 21 theme αMan 774:777 arg1 disaccharide 779:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 7 22 theme αMan 1562:1565 arg1 scaffold 1567:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 6 23 theme αMan-based 1173:1182 arg1 A 1190:1190 αGlcN(1↔1)αMan-based Lipid A 1163:1190 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) 1163:1212 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 24 theme NF-κB 1243:1247 arg1 signaling 1249:1257 NF-κB signaling 1243:1257 NF-κB signaling 1243:1257 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 3 25 theme 1↔1 770:772 arg1 disaccharide 779:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 7 26 theme 1↔1 1558:1560 arg1 scaffold 1567:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 2 27 theme restricted 457:466 arg1 mimetics 476:483 conformationally restricted Lipid A mimetics 440:483 conformationally restricted Lipid A mimetics 440:483 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 28 theme 1↔1 1169:1171 arg1 A 1190:1190 αGlcN(1↔1)αMan-based Lipid A 1163:1190 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) 1163:1212 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 2 29 theme coli 658:661 arg1 A 669:669 TLR4·MD-2-bound E. coli Lipid A 639:669 TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 639:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 5 30 with phosphorylation 1047:1061 arg1 acids 1026:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 3 31 theme convergent 707:716 arg1 route 728:732 A convergent synthetic route 705:732 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide 705:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 6 32 theme dendritic 1364:1372 arg1 cells 1374:1378 dendritic cells 1364:1378 dendritic cells 1364:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 5 33 theme global 1075:1080 arg1 deprotection 1082:1093 global deprotection 1075:1093 global deprotection 1075:1093 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 7 34 theme αGlcN 1552:1556 arg1 scaffold 1567:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 4 35 theme α-GlcN-lactol 896:908 arg1 glycosylation 855:867 the glycosylation 851:867 the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose 851:969 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 2 36 theme TLR4·MD-2-bound 639:653 arg1 A 669:669 TLR4·MD-2-bound E. coli Lipid A 639:669 TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 639:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 3 37 theme αGlcN 764:768 arg1 disaccharide 779:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 0 38 theme novel 58:62 arg1 class 64:68 a novel class 56:68 a novel class of potent Toll-like receptor 4 agonists 56:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 5 39 theme successive 1036:1045 arg1 phosphorylation 1047:1061 successive phosphorylation 1036:1061 successive phosphorylation followed by global deprotection 1036:1093 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 2 40 theme A 474:474 arg1 mimetics 476:483 conformationally restricted Lipid A mimetics 440:483 conformationally restricted Lipid A mimetics 440:483 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 2 41 theme trehalose-like 566:579 arg1 scaffold 596:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold 558:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 558:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 42 theme LPS-like 1315:1322 arg1 expression 1334:1343 robust LPS-like cytokines expression 1308:1343 robust LPS-like cytokines expression in macrophages and dendritic cells 1308:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 1 43 theme Toll-like 223:231 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 7 44 theme adjustable 1629:1638 arg1 activation 1645:1654 powerful and adjustable TLR4 activation 1616:1654 powerful and adjustable TLR4 activation 1616:1654 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 2 45 theme structural 366:375 arg1 requirements 377:388 the structural requirements 362:388 the structural requirements for TLR4·MD-2-specific ligands 362:419 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 0 46 theme αGlcN 15:19 arg1 Development 0:10 Development 0:10 Development of αGlcN(1↔1) 0:24 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 7 47 theme agonistic 1508:1516 arg1 conformation 1519:1530 the "agonistic" conformation 1503:1530 the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold 1503:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 7 48 theme powerful 1616:1623 arg1 activation 1645:1654 powerful and adjustable TLR4 activation 1616:1654 powerful and adjustable TLR4 activation 1616:1654 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 1 49 theme TLR4 245:248 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 4 50 theme 2-O-methylated 948:961 arg1 mannose 963:969 2-O-methylated mannose 948:969 2-O-methylated mannose 948:969 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 0 51 theme A 42:42 arg1 mimetics 44:51 lipid A mimetics 36:51 lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists 36:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 5 52 theme hexaacylated 1132:1143 arg1 A 1151:1151 monophosphorylated hexaacylated Lipid A 1113:1151 monophosphorylated hexaacylated Lipid A mimetics 1113:1160 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 1 53 theme factor 275:280 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 7 54 theme backbone 1491:1498 arg1 shape 1465:1469 the molecular shape 1451:1469 the molecular shape of its carbohydrate backbone 1451:1498 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 1 55 theme Lipid 182:186 arg1 A 188:188 a glycophospholipid Lipid A 162:188 a glycophospholipid Lipid A 162:188 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 1 55 theme Lipid 182:186 arg1 portion 125:131 The endotoxic portion 111:131 The endotoxic portion of lipopolysaccharide (LPS) 111:159 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 2 56 theme A 539:539 arg1 backbone 521:528 the flexible βGlcN(1→6)GlcN backbone 493:528 the flexible βGlcN(1→6)GlcN backbone of Lipid A 493:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 57 theme potent 1222:1227 arg1 activation 1229:1238 potent activation 1222:1238 potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells 1222:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 7 58 theme Lipid 1433:1437 arg1 A 1439:1439 Lipid A 1433:1439 Lipid A 1433:1439 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 7 59 theme conformational 1403:1416 arg1 flexibility 1418:1428 the conformational flexibility 1399:1428 the conformational flexibility of Lipid A 1399:1439 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 1 60 theme -myeloid 250:257 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 6 61 theme robust 1308:1313 arg1 expression 1334:1343 robust LPS-like cytokines expression 1308:1343 robust LPS-like cytokines expression in macrophages and dendritic cells 1308:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 2 62 theme βGlcN 506:510 arg1 backbone 521:528 the flexible βGlcN(1→6)GlcN backbone 493:528 the flexible βGlcN(1→6)GlcN backbone of Lipid A 493:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 0 63 theme agonists 101:108 arg1 class 64:68 a novel class 56:68 a novel class of potent Toll-like receptor 4 agonists 56:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 5 64 theme fatty 1020:1024 arg1 acids 1026:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 (R)-3-acyloxyacyl fatty acids 1002:1030 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 1 65 theme differentiation 259:273 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 6 66 theme hTLR4/hMD-2/CD14-transfected 1262:1289 arg1 cells 1298:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 1 67 theme immune 334:339 arg1 signaling 341:349 pro-inflammatory immune signaling 317:349 pro-inflammatory immune signaling 317:349 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 0 68 theme receptor 90:97 arg1 agonists 101:108 potent Toll-like receptor 4 agonists 73:108 potent Toll-like receptor 4 agonists 73:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 6 69 theme Lipid 1184:1188 arg1 A 1190:1190 αGlcN(1↔1)αMan-based Lipid A 1163:1190 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) 1163:1212 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 2 70 theme molecular 620:628 arg1 shape 630:634 the molecular shape 616:634 the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 616:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 71 theme signaling 1249:1257 arg1 expression 1334:1343 robust LPS-like cytokines expression 1308:1343 robust LPS-like cytokines expression in macrophages and dendritic cells 1308:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 71 theme signaling 1249:1257 arg1 activation 1229:1238 potent activation 1222:1238 potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells 1222:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 2 72 theme 1→6 512:514 arg1 backbone 521:528 the flexible βGlcN(1→6)GlcN backbone 493:528 the flexible βGlcN(1→6)GlcN backbone of Lipid A 493:539 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 2 73 theme Lipid 663:667 arg1 A 669:669 TLR4·MD-2-bound E. coli Lipid A 639:669 TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 639:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 7 74 theme rigid 1546:1550 arg1 scaffold 1567:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 a rigid αGlcN(1↔1)αMan scaffold 1544:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 2 75 theme E. 655:656 arg1 A 669:669 TLR4·MD-2-bound E. coli Lipid A 639:669 TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 639:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 3 76 theme synthetic 718:726 arg1 route 728:732 A convergent synthetic route 705:732 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide 705:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 1 77 theme endotoxic 115:123 arg1 A 188:188 a glycophospholipid Lipid A 162:188 a glycophospholipid Lipid A 162:188 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 1 77 theme endotoxic 115:123 arg1 portion 125:131 The endotoxic portion 111:131 The endotoxic portion of lipopolysaccharide (LPS) 111:159 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 4 78 theme 2-N-carbamate-protected 872:894 arg1 α-GlcN-lactol 896:908 2-N-carbamate-protected α-GlcN-lactol 872:908 2-N-carbamate-protected α-GlcN-lactol 872:908 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 2 79 theme 1↔1 587:589 arg1 scaffold 596:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold 558:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 558:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 80 dep mimetics 1192:1199 arg1 α-GM-LAM 1204:1211 α-GM-LAM 1204:1211 α-GM-LAM 1204:1211 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 80 dep mimetics 1192:1199 arg1 α 1202:1202 α 1202:1202 α 1202:1202 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 4 81 dep linkage 827:833 arg1 α- 819:820 The α,α-(1↔1) linkage 813:833 α- 819:820 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 1 82 theme MD-2 285:288 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 4 83 gly glycosylation 855:867 arg1 α-GlcN-lactol 896:908 2-N-carbamate-protected α-GlcN-lactol 872:908 2-N-carbamate-protected α-GlcN-lactol 872:908 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 2 84 theme αGlcN 581:585 arg1 scaffold 596:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold 558:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 558:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 3 85 theme protected 754:762 arg1 disaccharide 779:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 orthogonally protected αGlcN(1↔1)αMan disaccharide 741:790 A convergent synthetic route toward orthogonally protected αGlcN(1↔1)αMan disaccharide has been elaborated. 25252784 0 86 theme potent 73:78 arg1 receptor 90:97 potent Toll-like receptor 4 73:99 potent Toll-like receptor 4 agonists 73:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 1 87 theme complex 291:297 arg1 activation 205:214 the activation 201:214 the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling 201:349 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 2 88 theme Lipid 468:472 arg1 mimetics 476:483 conformationally restricted Lipid A mimetics 440:483 conformationally restricted Lipid A mimetics 440:483 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 6 89 theme cytokines 1324:1332 arg1 expression 1334:1343 robust LPS-like cytokines expression 1308:1343 robust LPS-like cytokines expression in macrophages and dendritic cells 1308:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 90 from expression 1334:1343 arg1 cells 1298:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 90 from expression 1334:1343 arg1 macrophages 1348:1358 macrophages 1348:1358 macrophages 1348:1358 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 90 from expression 1334:1343 arg1 cells 1374:1378 dendritic cells 1364:1378 dendritic cells 1364:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 4 91 theme mannose 963:969 arg1 N-phenyl-trifluoroacetimidate 915:943 N-phenyl-trifluoroacetimidate 915:943 N-phenyl-trifluoroacetimidate of 2-O-methylated mannose 915:969 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 2 92 theme αMan 591:594 arg1 scaffold 596:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold 558:603 a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure 558:702 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 1 93 theme Receptor 233:240 arg1 complex 291:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex 219:297 The endotoxic portion of lipopolysaccharide (LPS), a glycophospholipid Lipid A, initiates the activation of the Toll-like Receptor 4 (TLR4)-myeloid differentiation factor 2 (MD-2) complex, which results in pro-inflammatory immune signaling. 25252784 7 94 theme TLR4 1640:1643 arg1 activation 1645:1654 powerful and adjustable TLR4 activation 1616:1654 powerful and adjustable TLR4 activation 1616:1654 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 7 95 theme " 1517:1517 arg1 conformation 1519:1530 the "agonistic" conformation 1503:1530 the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold 1503:1574 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 0 96 theme lipid 36:40 arg1 A 42:42 lipid A 36:42 lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists 36:108 Development of αGlcN(1↔1)αMan-based lipid A mimetics as a novel class of potent Toll-like receptor 4 agonists. 25252784 5 97 theme monophosphorylated 1113:1130 arg1 A 1151:1151 monophosphorylated hexaacylated Lipid A 1113:1151 monophosphorylated hexaacylated Lipid A mimetics 1113:1160 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 5 98 theme Regioselective 972:985 arg1 acylation 987:995 Regioselective acylation 972:995 Regioselective acylation with (R)-3-acyloxyacyl fatty acids 972:1030 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 2 99 theme TLR4·MD-2-specific 394:411 arg1 ligands 413:419 TLR4·MD-2-specific ligands 394:419 TLR4·MD-2-specific ligands 394:419 To unveil the structural requirements for TLR4·MD-2-specific ligands, we have developed conformationally restricted Lipid A mimetics wherein the flexible βGlcN(1→6)GlcN backbone of Lipid A is exchanged for a rigid trehalose-like αGlcN(1↔1)αMan scaffold resembling the molecular shape of TLR4·MD-2-bound E. coli Lipid A disclosed in the X-ray structure. 25252784 5 100 theme Lipid 1145:1149 arg1 A 1151:1151 monophosphorylated hexaacylated Lipid A 1113:1151 monophosphorylated hexaacylated Lipid A mimetics 1113:1160 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 4 101 with glycosylation 855:867 arg1 N-phenyl-trifluoroacetimidate 915:943 N-phenyl-trifluoroacetimidate 915:943 N-phenyl-trifluoroacetimidate of 2-O-methylated mannose 915:969 The α,α-(1↔1) linkage was attained by the glycosylation of 2-N-carbamate-protected α-GlcN-lactol with N-phenyl-trifluoroacetimidate of 2-O-methylated mannose. 25252784 7 102 theme carbohydrate 1478:1489 arg1 backbone 1491:1498 its carbohydrate backbone 1474:1498 its carbohydrate backbone 1474:1498 Thus, restricting the conformational flexibility of Lipid A by fixing the molecular shape of its carbohydrate backbone in the "agonistic" conformation attained by a rigid αGlcN(1↔1)αMan scaffold represents an efficient approach toward powerful and adjustable TLR4 activation. 25252784 5 103 dep -3-acyloxyacyl 1005:1018 arg1 R 1003:1003 R 1003:1003 R 1003:1003 Regioselective acylation with (R)-3-acyloxyacyl fatty acids and successive phosphorylation followed by global deprotection afforded bis- and monophosphorylated hexaacylated Lipid A mimetics. 25252784 6 104 from activation 1229:1238 arg1 cells 1298:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 hTLR4/hMD-2/CD14-transfected HEK293 cells 1262:1302 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 104 from activation 1229:1238 arg1 macrophages 1348:1358 macrophages 1348:1358 macrophages 1348:1358 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25252784 6 104 from activation 1229:1238 arg1 cells 1374:1378 dendritic cells 1364:1378 dendritic cells 1364:1378 αGlcN(1↔1)αMan-based Lipid A mimetics (α,α-GM-LAM) induced potent activation of NF-κB signaling in hTLR4/hMD-2/CD14-transfected HEK293 cells and robust LPS-like cytokines expression in macrophages and dendritic cells. 25291402 3 0 from bottlenecks 444:454 arg1 synthesis 472:480 the chemical synthesis 459:480 the chemical synthesis of complex glycans 459:499 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 7 1 theme derivatives 1308:1318 arg1 glycosylation 1278:1290 the glycosylation 1274:1290 the glycosylation of persilylated derivatives, all of which were conducted in a single vessel 1274:1366 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 7 1 theme derivatives 1308:1318 arg1 number 1245:1250 a number 1243:1250 a number of thioglucosides 1243:1268 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 1 2 theme information 191:201 arg1 density 203:209 their large structural information density 168:209 their large structural information density 168:209 Living organisms employ glycans as recognition elements because of their large structural information density. 25291402 4 3 theme building-block 619:632 arg1 acquisition 634:644 building-block acquisition 619:644 building-block acquisition 619:644 Thus, easy, rapid, and efficient methods for building-block acquisition are desirable. 25291402 7 4 theme persilylated 1295:1306 arg1 derivatives 1308:1318 persilylated derivatives 1295:1318 persilylated derivatives 1295:1318 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 3 5 theme blocks 566:571 arg1 preparation 508:518 the preparation 504:518 the preparation of suitably protected monosaccharide building blocks 504:571 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 6 6 dep in 842:843 arg1 situ 845:848 situ 845:848 situ 845:848 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 5 7 theme free 715:718 arg1 sugars 720:725 the free sugars 711:725 the free sugars 711:725 Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. 25291402 3 8 theme building 557:564 arg1 blocks 566:571 suitably protected monosaccharide building blocks 523:571 suitably protected monosaccharide building blocks 523:571 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 2 9 theme natural 390:396 arg1 sources 398:404 natural sources 390:404 natural sources 390:404 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 6 10 theme intermediates 904:916 arg1 generation 850:859 the in situ generation 838:859 the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation 838:1127 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 1 11 theme Living 101:106 arg1 organisms 108:116 Living organisms 101:116 Living organisms 101:116 Living organisms employ glycans as recognition elements because of their large structural information density. 25291402 7 12 gly glycosylation 1278:1290 arg1 thioglucosides 1255:1268 thioglucosides 1255:1268 thioglucosides 1255:1268 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 7 12 gly glycosylation 1278:1290 arg1 derivatives 1308:1318 persilylated derivatives 1295:1318 persilylated derivatives 1295:1318 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 6 13 theme monosaccharide 889:902 arg1 intermediates 904:916 per-O-trimethylsilylated monosaccharide intermediates 864:916 per-O-trimethylsilylated monosaccharide intermediates 864:916 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 7 14 theme single 1354:1359 arg1 vessel 1361:1366 a single vessel 1352:1366 a single vessel 1352:1366 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 3 15 theme complex 485:491 arg1 glycans 493:499 complex glycans 485:499 complex glycans 485:499 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 0 16 theme one-pot 19:25 arg1 synthesis 27:35 Microwave-assisted one-pot synthesis 0:35 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 0:99 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 25291402 7 17 theme thioglucosides 1255:1268 arg1 glycosylation 1278:1290 the glycosylation 1274:1290 the glycosylation of persilylated derivatives, all of which were conducted in a single vessel 1274:1366 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 7 17 theme thioglucosides 1255:1268 arg1 number 1245:1250 a number 1243:1250 a number of thioglucosides 1243:1268 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 6 18 theme per-O-trimethylsilylated 864:887 arg1 intermediates 904:916 per-O-trimethylsilylated monosaccharide intermediates 864:916 per-O-trimethylsilylated monosaccharide intermediates 864:916 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 3 19 theme chemical 463:470 arg1 synthesis 472:480 the chemical synthesis 459:480 the chemical synthesis of complex glycans 459:499 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 0 20 theme Microwave-assisted 0:17 arg1 synthesis 27:35 Microwave-assisted one-pot synthesis 0:35 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 0:99 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 25291402 8 21 theme stages 1560:1565 arg1 number 1537:1542 minimal number 1529:1542 minimal number of purification stages 1529:1565 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 8 22 theme purification 1547:1558 arg1 stages 1560:1565 purification stages 1547:1565 purification stages 1547:1565 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 8 23 theme developed 1375:1383 arg1 approaches 1385:1394 These developed approaches 1369:1394 These developed approaches 1369:1394 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 8 24 theme oligosaccharide 1481:1495 arg1 assembly 1497:1504 oligosaccharide assembly 1481:1504 oligosaccharide assembly 1481:1504 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 2 25 theme compounds 348:356 arg1 quantities 328:337 sufficient quantities 317:337 sufficient quantities of these compounds 317:356 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 7 26 theme protecting 1189:1198 arg1 installation 1206:1217 regioselective protecting group installation 1174:1217 regioselective protecting group installation 1174:1217 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 6 27 theme microwave 1107:1115 arg1 irradiation 1117:1127 microwave irradiation 1107:1127 microwave irradiation 1107:1127 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 2 28 theme sufficient 317:326 arg1 quantities 328:337 sufficient quantities 317:337 sufficient quantities of these compounds 317:356 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 6 29 theme trimethylsilyl 997:1010 arg1 trifluoromethanesulfonate 1012:1036 trimethylsilyl trifluoromethanesulfonate 997:1036 trimethylsilyl trifluoromethanesulfonate 997:1036 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 1 30 theme recognition 136:146 arg1 glycans 125:131 glycans 125:131 glycans 125:131 Living organisms employ glycans as recognition elements because of their large structural information density. 25291402 1 30 theme recognition 136:146 arg1 elements 148:155 recognition elements 136:155 recognition elements 136:155 Living organisms employ glycans as recognition elements because of their large structural information density. 25291402 8 31 theme short 1511:1515 arg1 period 1517:1522 a short period 1509:1522 a short period with minimal number of purification stages 1509:1565 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 4 32 theme efficient 597:605 arg1 methods 607:613 easy, rapid, and efficient methods 580:613 easy, rapid, and efficient methods for building-block acquisition 580:644 Thus, easy, rapid, and efficient methods for building-block acquisition are desirable. 25291402 5 33 theme microwave-assisted 777:794 arg1 synthesis 804:812 microwave-assisted one-pot synthesis 777:812 microwave-assisted one-pot synthesis 777:812 Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. 25291402 2 34 theme sugar 225:229 arg1 structures 231:240 Well-defined sugar structures 212:240 Well-defined sugar structures 212:240 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 3 35 theme glycans 493:499 arg1 synthesis 472:480 the chemical synthesis 459:480 the chemical synthesis of complex glycans 459:499 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 0 36 theme 1,6-anhydrosugars 40:56 arg1 synthesis 27:35 Microwave-assisted one-pot synthesis 0:35 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 0:99 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 25291402 4 37 theme easy 580:583 arg1 methods 607:613 easy, rapid, and efficient methods 580:613 easy, rapid, and efficient methods for building-block acquisition 580:644 Thus, easy, rapid, and efficient methods for building-block acquisition are desirable. 25291402 5 38 theme one-pot 796:802 arg1 synthesis 804:812 microwave-assisted one-pot synthesis 777:812 microwave-assisted one-pot synthesis 777:812 Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. 25291402 2 39 theme Well-defined 212:223 arg1 structures 231:240 Well-defined sugar structures 212:240 Well-defined sugar structures 212:240 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 7 40 theme regioselective 1174:1187 arg1 installation 1206:1217 regioselective protecting group installation 1174:1217 regioselective protecting group installation 1174:1217 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 8 41 theme building 1461:1468 arg1 blocks 1470:1475 suitably protected building blocks 1442:1475 suitably protected building blocks for oligosaccharide assembly 1442:1504 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 0 42 theme protected 75:83 arg1 thioglycosides 85:98 orthogonally protected thioglycosides 62:98 orthogonally protected thioglycosides 62:98 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 25291402 6 43 theme in 842:843 arg1 generation 850:859 the in situ generation 838:859 the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation 838:1127 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 3 44 theme protected 532:540 arg1 blocks 566:571 suitably protected monosaccharide building blocks 523:571 suitably protected monosaccharide building blocks 523:571 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 4 45 theme rapid 586:590 arg1 methods 607:613 easy, rapid, and efficient methods 580:613 easy, rapid, and efficient methods for building-block acquisition 580:644 Thus, easy, rapid, and efficient methods for building-block acquisition are desirable. 25291402 7 46 theme group 1200:1204 arg1 installation 1206:1217 regioselective protecting group installation 1174:1217 regioselective protecting group installation 1174:1217 We successfully extended the methodology to regioselective protecting group installation and manipulation toward a number of thioglucosides and the glycosylation of persilylated derivatives, all of which were conducted in a single vessel. 25291402 3 47 theme monosaccharide 542:555 arg1 blocks 566:571 suitably protected monosaccharide building blocks 523:571 suitably protected monosaccharide building blocks 523:571 Among the bottlenecks in the chemical synthesis of complex glycans is the preparation of suitably protected monosaccharide building blocks. 25291402 5 48 theme notable 734:740 arg1 derivatives 757:767 notable monosaccharide derivatives 734:767 notable monosaccharide derivatives 734:767 Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. 25291402 8 49 with period 1517:1522 arg1 number 1537:1542 minimal number 1529:1542 minimal number of purification stages 1529:1565 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 5 50 theme monosaccharide 742:755 arg1 derivatives 757:767 notable monosaccharide derivatives 734:767 notable monosaccharide derivatives 734:767 Herein, we describe routes directly starting from the free sugars toward notable monosaccharide derivatives through microwave-assisted one-pot synthesis. 25291402 2 51 theme functions 302:310 arg1 advantage 282:290 advantage 282:290 advantage of glycan functions 282:310 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 8 52 theme minimal 1529:1535 arg1 number 1537:1542 minimal number 1529:1542 minimal number of purification stages 1529:1565 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 2 53 theme glycan 295:300 arg1 functions 302:310 glycan functions 295:310 glycan functions 295:310 Well-defined sugar structures are needed to fully understand and take advantage of glycan functions, but sufficient quantities of these compounds cannot be readily obtained from natural sources and have to be synthesized. 25291402 8 54 theme protected 1451:1459 arg1 blocks 1470:1475 suitably protected building blocks 1442:1475 suitably protected building blocks for oligosaccharide assembly 1442:1504 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 1 55 theme large 174:178 arg1 information 191:201 their large structural information 168:201 their large structural information density 168:209 Living organisms employ glycans as recognition elements because of their large structural information density. 25291402 6 56 with treatment 975:983 arg1 trifluoromethanesulfonate 1012:1036 trimethylsilyl trifluoromethanesulfonate 997:1036 trimethylsilyl trifluoromethanesulfonate 997:1036 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 6 56 with treatment 975:983 arg1 silane 1070:1075 silane 1070:1075 silane 1070:1075 The procedure followed the in situ generation of per-O-trimethylsilylated monosaccharide intermediates, which provided 1,6-anhydrosugars or thioglycosides upon treatment with either trimethylsilyl trifluoromethanesulfonate or trimethyl(4-methylphenylthio)silane and ZnI2, respectively, under microwave irradiation. 25291402 0 57 theme thioglycosides 85:98 arg1 synthesis 27:35 Microwave-assisted one-pot synthesis 0:35 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 0:99 Microwave-assisted one-pot synthesis of 1,6-anhydrosugars and orthogonally protected thioglycosides. 25291402 8 58 theme blocks 1470:1475 arg1 arrays 1432:1437 arrays 1432:1437 arrays of suitably protected building blocks for oligosaccharide assembly 1432:1504 These developed approaches open the possibility for generating arrays of suitably protected building blocks for oligosaccharide assembly in a short period with minimal number of purification stages. 25291402 1 59 theme structural 180:189 arg1 information 191:201 their large structural information 168:201 their large structural information density 168:209 Living organisms employ glycans as recognition elements because of their large structural information density. 26802542 6 0 theme GH 1312:1313 arg1 β-galactosidases 1317:1332 both thermostable GH 1 β-galactosidases 1294:1332 both thermostable GH 1 β-galactosidases 1294:1332 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 3 1 dep LNnT 638:641 arg1 LacNAc 668:673 LacNAc 668:673 LacNAc; Gal-β(1,4)-GlcNAc 668:692 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 7 2 theme reaction 1611:1618 arg1 yields 1620:1625 5-6 times higher reaction yields 1594:1625 5-6 times higher reaction yields 1594:1625 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 1 3 theme lactose-based 154:166 arg1 molecules 168:176 bioactive lactose-based molecules 144:176 bioactive lactose-based molecules present in human breast milk 144:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 5 4 from activity 1183:1190 arg1 formation 1199:1207 the formation 1195:1207 the formation of LacNAc and LNnT 1195:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 4 5 from circulans 996:1004 arg1 BgaD-D 925:930 BgaD-D 925:930 BgaD-D 925:930 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 5 from circulans 996:1004 arg1 β-galactosidase 966:980 the GH 42 β-galactosidase 956:980 the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 956:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 5 from circulans 996:1004 arg1 version 945:951 a truncated version 933:951 a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 933:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 1 6 theme molecules 168:176 arg1 family 134:139 a unique family 125:139 a unique family of bioactive lactose-based molecules present in human breast milk 125:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 4 7 theme β-galactosidase 966:980 arg1 BgaD-D 925:930 BgaD-D 925:930 BgaD-D 925:930 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 7 theme β-galactosidase 966:980 arg1 version 945:951 a truncated version 933:951 a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 933:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 7 8 theme thermal 1428:1434 arg1 stability 1436:1444 The thermal stability 1424:1444 The thermal stability of the three β-galactosidases 1424:1474 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 7 9 from efficiency 1542:1551 arg1 formation 1560:1568 the formation 1556:1568 the formation of LacNAc 1556:1578 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 5 10 from formation 1199:1207 arg1 terms 1147:1151 terms 1147:1151 terms of their transgalactosylation activity in the formation of LacNAc and LNnT 1147:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 5 11 theme LacNAc 1212:1217 arg1 formation 1199:1207 the formation 1195:1207 the formation of LacNAc and LNnT 1195:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 4 12 theme substrate 1056:1064 arg1 concentrations 1066:1079 low substrate concentrations 1052:1079 low substrate concentrations 1052:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 5 13 from terms 1147:1151 arg1 formation 1199:1207 the formation 1195:1207 the formation of LacNAc and LNnT 1195:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 6 14 from transgalactosidase 1270:1287 arg1 cases 1236:1240 all cases 1232:1240 all cases 1232:1240 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 4 15 theme hyperthermostable 743:759 arg1 Ttβ-gly 784:790 Ttβ-gly 784:790 Ttβ-gly 784:790 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 15 theme hyperthermostable 743:759 arg1 CelB 827:830 CelB 827:830 CelB 827:830 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 15 theme hyperthermostable 743:759 arg1 β-galactosidases 766:781 Two hyperthermostable GH 1 β-galactosidases 739:781 Two hyperthermostable GH 1 β-galactosidases 739:781 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 3 16 theme core 715:718 arg1 latter 700:705 latter 700:705 latter 700:705 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 3 16 theme core 715:718 arg1 structure 720:728 a core structure 713:728 a core structure in HMOs 713:736 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 1 17 theme Human 80:84 arg1 HMOs 109:112 HMOs 109:112 HMOs 109:112 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 1 17 theme Human 80:84 arg1 oligosaccharides 91:106 Human milk oligosaccharides 80:106 Human milk oligosaccharides (HMOs) 80:113 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 4 18 theme low 1052:1054 arg1 concentrations 1066:1079 low substrate concentrations 1052:1079 low substrate concentrations 1052:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 1 19 theme present 178:184 arg1 molecules 168:176 bioactive lactose-based molecules 144:176 bioactive lactose-based molecules present in human breast milk 144:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 7 20 theme reaction 1653:1660 arg1 times 1662:1666 significantly shorter reaction times 1631:1666 significantly shorter reaction times 1631:1666 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 3 21 theme N-acetyllactosamine 647:665 arg1 synthesis 625:633 synthesis 625:633 synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc) 625:693 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 1 22 theme milk 86:89 arg1 HMOs 109:112 HMOs 109:112 HMOs 109:112 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 1 22 theme milk 86:89 arg1 oligosaccharides 91:106 Human milk oligosaccharides 80:106 Human milk oligosaccharides (HMOs) 80:113 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 6 23 theme potent 1263:1268 arg1 transgalactosidase 1270:1287 the most potent transgalactosidase 1254:1287 the most potent transgalactosidase 1254:1287 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 6 23 theme potent 1263:1268 arg1 BgaD-D 1243:1248 BgaD-D 1243:1248 BgaD-D 1243:1248 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 2 24 theme 1,4 382:384 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 4 25 theme truncated 935:943 arg1 BgaD-D 925:930 BgaD-D 925:930 BgaD-D 925:930 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 25 theme truncated 935:943 arg1 version 945:951 a truncated version 933:951 a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 933:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 5 26 theme current 1130:1136 arg1 study 1138:1142 the current study 1126:1142 the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT 1126:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 7 27 theme higher 1604:1609 arg1 yields 1620:1625 5-6 times higher reaction yields 1594:1625 5-6 times higher reaction yields 1594:1625 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 1 28 theme human 189:193 arg1 milk 202:205 human breast milk 189:205 human breast milk 189:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 2 29 theme 1,4 407:409 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 3 30 theme 1,4 682:684 arg1 -GlcNAc 686:692 Gal-β(1,4)-GlcNAc 676:692 LacNAc; Gal-β(1,4)-GlcNAc 668:692 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 4 31 theme GH 761:762 arg1 Ttβ-gly 784:790 Ttβ-gly 784:790 Ttβ-gly 784:790 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 31 theme GH 761:762 arg1 CelB 827:830 CelB 827:830 CelB 827:830 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 31 theme GH 761:762 arg1 β-galactosidases 766:781 Two hyperthermostable GH 1 β-galactosidases 739:781 Two hyperthermostable GH 1 β-galactosidases 739:781 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 7 32 theme reaction 1533:1540 arg1 efficiency 1542:1551 the reaction efficiency 1529:1551 the reaction efficiency in the formation of LacNAc 1529:1578 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 4 33 theme transgalactosylation 1019:1038 arg1 activity 1040:1047 high transgalactosylation activity 1014:1047 high transgalactosylation activity 1014:1047 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 2 34 theme Gal-β 376:380 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 4 35 from version 945:951 arg1 circulans 996:1004 Bacillus circulans 987:1004 Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 987:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 3 36 from structure 720:728 arg1 HMOs 733:736 HMOs 733:736 HMOs 733:736 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 6 37 theme thermostable 1299:1310 arg1 β-galactosidases 1317:1332 both thermostable GH 1 β-galactosidases 1294:1332 both thermostable GH 1 β-galactosidases 1294:1332 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 4 38 theme high 1014:1017 arg1 activity 1040:1047 high transgalactosylation activity 1014:1047 high transgalactosylation activity 1014:1047 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 2 39 theme -Gal-β 400:405 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 3 40 theme Gal-β 676:680 arg1 -GlcNAc 686:692 Gal-β(1,4)-GlcNAc 676:692 LacNAc; Gal-β(1,4)-GlcNAc 668:692 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 7 41 theme β-galactosidases 1459:1474 arg1 stability 1436:1444 The thermal stability 1424:1444 The thermal stability of the three β-galactosidases 1424:1474 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 2 42 theme 1,3 396:398 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 3 43 theme reaction 437:444 arg1 times 446:450 reaction times 437:450 reaction times 437:450 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 5 44 from study 1138:1142 arg1 terms 1147:1151 terms 1147:1151 terms of their transgalactosylation activity in the formation of LacNAc and LNnT 1147:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 1 45 attach present 178:184 arg2 molecules 168:176 bioactive lactose-based molecules 144:176 bioactive lactose-based molecules present in human breast milk 144:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 1 45 attach present 178:184 arg1 milk 202:205 human breast milk 189:205 human breast milk 189:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 3 46 theme microbial 517:525 arg1 growth 527:532 microbial growth 517:532 microbial growth 517:532 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 1 47 theme breast 195:200 arg1 milk 202:205 human breast milk 189:205 human breast milk 189:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 7 48 theme shorter 1645:1651 arg1 times 1662:1666 significantly shorter reaction times 1631:1666 significantly shorter reaction times 1631:1666 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 4 49 theme Pyrococcus 837:846 arg1 furiosus 848:855 Pyrococcus furiosus 837:855 Pyrococcus furiosus 837:855 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 2 50 theme cheap 227:231 arg1 donor 233:237 a cheap donor 225:237 a cheap donor 225:237 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 2 50 theme cheap 227:231 arg1 lactose 214:220 lactose 214:220 lactose 214:220 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 2 51 theme -GlcNAc-β 386:394 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 0 52 theme milk 57:60 arg1 oligosaccharides 62:77 human milk oligosaccharides 51:77 human milk oligosaccharides 51:77 Thermostable β-galactosidases for the synthesis of human milk oligosaccharides. 26802542 4 53 theme thermophilus 805:816 arg1 HB27 818:821 Thermus thermophilus HB27 797:821 Thermus thermophilus HB27 797:821 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 7 54 theme LacNAc 1573:1578 arg1 formation 1560:1568 the formation 1556:1568 the formation of LacNAc 1556:1578 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 6 55 theme LacNAc 1371:1376 arg1 formation 1349:1357 formation 1349:1357 formation of LNnT and LacNAc 1349:1376 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 2 56 theme oligosaccharide 333:347 arg1 LNnT 370:373 LNnT 370:373 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 2 56 theme oligosaccharide 333:347 arg1 lacto-N-neotetraose 349:367 the human milk oligosaccharide lacto-N-neotetraose 318:367 the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc) 318:415 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 0 57 theme human 51:55 arg1 oligosaccharides 62:77 human milk oligosaccharides 51:77 human milk oligosaccharides 51:77 Thermostable β-galactosidases for the synthesis of human milk oligosaccharides. 26802542 3 58 dep LacNAc 668:673 arg1 -GlcNAc 686:692 Gal-β(1,4)-GlcNAc 676:692 LacNAc; Gal-β(1,4)-GlcNAc 668:692 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 4 59 theme Thermus 797:803 arg1 HB27 818:821 Thermus thermophilus HB27 797:821 Thermus thermophilus HB27 797:821 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 1 60 from milk 202:205 arg1 present 178:184 present 178:184 present 178:184 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 2 61 theme milk 328:331 arg1 LNnT 370:373 LNnT 370:373 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 2 61 theme milk 328:331 arg1 lacto-N-neotetraose 349:367 the human milk oligosaccharide lacto-N-neotetraose 318:367 the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc) 318:415 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 5 62 theme transgalactosylation 1162:1181 arg1 activity 1183:1190 their transgalactosylation activity 1156:1190 their transgalactosylation activity in the formation of LacNAc and LNnT 1156:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 3 63 theme current 539:545 arg1 study 547:551 the current study 535:551 the current study 535:551 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 1 64 theme unique 127:132 arg1 family 134:139 a unique family 125:139 a unique family of bioactive lactose-based molecules present in human breast milk 125:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 5 65 theme LNnT 1223:1226 arg1 formation 1199:1207 the formation 1195:1207 the formation of LacNAc and LNnT 1195:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 2 66 dep LNnT 370:373 arg1 -Glc 411:414 Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 376:414 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 7 67 used used 1512:1515 arg2 results 1499:1505 the results 1495:1505 the results 1495:1505 The thermal stability of the three β-galactosidases was elucidated and the results were used to optimize the reaction efficiency in the formation of LacNAc, resulting in 5-6 times higher reaction yields and significantly shorter reaction times. 26802542 2 68 theme human 322:326 arg1 LNnT 370:373 LNnT 370:373 LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc 370:414 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 2 68 theme human 322:326 arg1 lacto-N-neotetraose 349:367 the human milk oligosaccharide lacto-N-neotetraose 318:367 the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc) 318:415 Using lactose as a cheap donor, some β-galactosidases (EC 3.2.1.23) can catalyze transgalactosylation to form the human milk oligosaccharide lacto-N-neotetraose (LNnT; Gal-β(1,4)-GlcNAc-β(1,3)-Gal-β(1,4)-Glc). 26802542 3 69 theme thermostable 591:602 arg1 β-galactosidases 604:619 thermostable β-galactosidases 591:619 thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc) 591:693 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 6 70 theme higher 1399:1404 arg1 yields 1406:1411 higher yields 1399:1411 higher yields than CelB 1399:1421 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 5 71 theme activity 1183:1190 arg1 terms 1147:1151 terms 1147:1151 terms of their transgalactosylation activity in the formation of LacNAc and LNnT 1147:1226 The three β-galactosidases were compared in the current study in terms of their transgalactosylation activity in the formation of LacNAc and LNnT. 26802542 0 72 theme oligosaccharides 62:77 arg1 synthesis 38:46 the synthesis 34:46 the synthesis of human milk oligosaccharides 34:77 Thermostable β-galactosidases for the synthesis of human milk oligosaccharides. 26802542 3 73 theme LNnT 638:641 arg1 synthesis 625:633 synthesis 625:633 synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc) 625:693 In order to reduce reaction times and be able to work at temperatures, which are less welcoming to microbial growth, the current study investigates the possibility of using thermostable β-galactosidases for synthesis of LNnT and N-acetyllactosamine (LacNAc; Gal-β(1,4)-GlcNAc), the latter being a core structure in HMOs. 26802542 6 74 theme LNnT 1362:1365 arg1 formation 1349:1357 formation 1349:1357 formation of LNnT and LacNAc 1349:1376 In all cases, BgaD-D was the most potent transgalactosidase, but both thermostable GH 1 β-galactosidases could catalyze formation of LNnT and LacNAc, with Ttβ-gly giving higher yields than CelB. 26802542 1 75 from present 178:184 arg1 milk 202:205 human breast milk 189:205 human breast milk 189:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 1 76 theme bioactive 144:152 arg1 molecules 168:176 bioactive lactose-based molecules 144:176 bioactive lactose-based molecules present in human breast milk 144:205 Human milk oligosaccharides (HMOs) designate a unique family of bioactive lactose-based molecules present in human breast milk. 26802542 4 77 theme GH 960:961 arg1 β-galactosidase 966:980 the GH 42 β-galactosidase 956:980 the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations 956:1079 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 26802542 4 78 from expression 883:892 arg1 coli 909:912 Escherichia coli 897:912 Escherichia coli 897:912 Two hyperthermostable GH 1 β-galactosidases, Ttβ-gly from Thermus thermophilus HB27 and CelB from Pyrococcus furiosus, were codon-optimized for expression in Escherichia coli along with BgaD-D, a truncated version of the GH 42 β-galactosidase from Bacillus circulans showing high transgalactosylation activity at low substrate concentrations. 25305020 2 0 with interaction 322:332 arg1 ECD 361:363 the ECD 357:363 the ECD of the receptor 357:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 4 1 theme CaSR 916:919 arg1 ECD 921:923 the CaSR ECD 912:923 the CaSR ECD 912:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 9 2 theme CaSR 1745:1748 arg1 properties 1731:1740 the structural properties 1716:1740 the structural properties of CaSR and other members of family C of the GPCR superfamily 1716:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 2 3 theme weak 591:594 arg1 interactions 596:607 weak interactions 591:607 weak interactions in solution 591:619 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 4 4 dep % 1014:1014 arg1 20 1012:1013 20 1012:1013 20 1012:1013 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 1 5 theme Ca 258:259 arg1 homeostasis 265:275 Ca(2+) homeostasis 258:275 Ca(2+) homeostasis 258:275 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 4 6 theme ∼ 995:995 arg1 % 999:999 % 999:999 % 999:999 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 6 7 with interactions 1249:1260 arg1 Ca 1267:1268 Ca 1267:1268 Ca(2+) 1267:1272 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 6 7 with interactions 1249:1260 arg1 2+ 1270:1271 2+ 1270:1271 2+ 1270:1271 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 1 8 theme extracellular 184:196 arg1 domain 198:203 the dimeric extracellular domain 172:203 the dimeric extracellular domain (ECD) of the CaSR 172:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 8 theme extracellular 184:196 arg1 CaSR 218:221 the CaSR 214:221 the CaSR 214:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 8 theme extracellular 184:196 arg1 ECD 206:208 ECD 206:208 ECD 206:208 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 2 9 theme high 514:517 arg1 degree 519:524 its high degree 510:524 its high degree of glycosylation 510:541 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 4 10 theme % 999:999 arg1 α-helix 1001:1007 ∼ 50% α-helix 995:1007 ∼ 50% α-helix 995:1007 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 2 11 from interactions 596:607 arg1 solution 612:619 solution 612:619 solution 612:619 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 6 12 theme specific 1302:1309 arg1 forms 1329:1333 specific (15)N Phe-labeled forms 1302:1333 specific (15)N Phe-labeled forms of the ECD 1302:1344 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 4 13 theme similar 961:967 arg1 structures 979:988 similar secondary structures 961:988 similar secondary structures 961:988 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 13 theme similar 961:967 arg1 dimers 942:947 dimers 942:947 dimers 942:947 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 13 theme similar 961:967 arg1 forms 903:907 Both glycosylated forms 885:907 Both glycosylated forms of the CaSR ECD 885:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 14 dep content 1024:1030 arg1 % 1014:1014 % 1014:1014 % 1014:1014 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 8 15 theme binding 1527:1533 arg1 affinity 1535:1542 the binding affinity 1523:1542 the binding affinity of the CaSR ECD for Ca(2+) 1523:1569 We further demonstrated that l-Phe increases the binding affinity of the CaSR ECD for Ca(2+). 25305020 7 16 dep CaSR 1458:1461 arg1 ECD 1463:1465 ECD 1463:1465 ECD 1463:1465 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 7 17 theme NMR 1378:1380 arg1 approaches 1382:1391 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 8 18 theme CaSR 1551:1554 arg1 ECD 1556:1558 the CaSR ECD 1547:1558 the CaSR ECD 1547:1558 We further demonstrated that l-Phe increases the binding affinity of the CaSR ECD for Ca(2+). 25305020 7 19 theme transfer 1358:1365 arg1 difference 1367:1376 Saturation transfer difference 1347:1376 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 3 20 theme extracellular 673:685 arg1 domain 687:692 the glycosylated extracellular domain 656:692 the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 656:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 20 theme extracellular 673:685 arg1 receptor 713:720 calcium-sensing receptor 697:720 calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 697:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 0 21 theme extracellular 62:74 arg1 receptor 106:113 the calcium-sensing receptor 86:113 the calcium-sensing receptor 86:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 0 21 theme extracellular 62:74 arg1 domain 76:81 the extracellular domain 58:81 the extracellular domain of the calcium-sensing receptor 58:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 2 22 with impact 399:404 arg1 ECD 361:363 the ECD 357:363 the ECD of the receptor 357:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 23 theme conformational 438:451 arg1 changes 453:459 associated conformational changes 427:459 associated conformational changes 427:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 3 24 theme receptor 713:720 arg1 domain 687:692 the glycosylated extracellular domain 656:692 the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 656:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 24 theme receptor 713:720 arg1 receptor 713:720 calcium-sensing receptor 697:720 calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 697:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 0 25 theme calcium-sensing 90:104 arg1 receptor 106:113 the calcium-sensing receptor 86:113 the calcium-sensing receptor 86:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 3 26 theme complex 772:778 arg1 structures 805:814 either complex or high mannose N-glycan structures 765:814 either complex or high mannose N-glycan structures 765:814 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 6 27 theme N 1315:1315 arg1 forms 1329:1333 specific (15)N Phe-labeled forms 1302:1333 specific (15)N Phe-labeled forms of the ECD 1302:1344 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 2 28 theme glycosylation 529:541 arg1 degree 519:524 its high degree 510:524 its high degree of glycosylation 510:541 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 28 theme glycosylation 529:541 arg1 lack 552:555 the lack 548:555 the lack of biophysical methods to monitor weak interactions in solution 548:619 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 28 theme glycosylation 529:541 arg1 size 493:496 the large size 483:496 the large size of the ECD 483:507 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 1 29 dep in 125:126 arg1 vivo 128:131 vivo 128:131 vivo 128:131 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 3 30 dep residues 736:743 arg1 20-612 745:750 20-612 745:750 20-612 745:750 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 2 31 theme biophysical 560:570 arg1 methods 572:578 biophysical methods 560:578 biophysical methods 560:578 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 0 32 theme Direct 0:5 arg1 determination 7:19 Direct determination 0:19 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor 0:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 3 33 theme N-glycan 796:803 arg1 structures 805:814 either complex or high mannose N-glycan structures 765:814 either complex or high mannose N-glycan structures 765:814 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 4 34 theme buried 1044:1049 arg1 environment 1055:1065 a well buried Trp environment 1037:1065 a well buried Trp environment 1037:1065 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 2 35 theme Phe 348:350 arg1 interaction 322:332 direct interaction 315:332 direct interaction of Ca(2+) and Phe with the ECD of the receptor 315:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 35 theme Phe 348:350 arg1 impact 399:404 the resultant impact 385:404 the resultant impact on its structure and associated conformational changes 385:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 6 36 theme proteins 1223:1230 arg1 changes 1208:1214 The local conformational changes 1183:1214 The local conformational changes of the proteins induced by their interactions with Ca(2+) 1183:1272 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 1 37 theme Numerous 116:123 arg1 studies 144:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 3 38 theme high 783:786 arg1 structures 805:814 either complex or high mannose N-glycan structures 765:814 either complex or high mannose N-glycan structures 765:814 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 0 39 theme ligand 33:38 arg1 interactions 40:51 multiple ligand interactions 24:51 multiple ligand interactions with the extracellular domain of the calcium-sensing receptor 24:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 2 40 theme receptor 372:379 arg1 ECD 361:363 the ECD 357:363 the ECD of the receptor 357:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 9 41 theme C 1778:1778 arg1 CaSR 1745:1748 CaSR 1745:1748 CaSR 1745:1748 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 9 41 theme C 1778:1778 arg1 members 1760:1766 other members 1754:1766 other members of family C of the GPCR superfamily 1754:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 6 42 theme conformational 1193:1206 arg1 changes 1208:1214 The local conformational changes 1183:1214 The local conformational changes of the proteins induced by their interactions with Ca(2+) 1183:1272 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 9 43 theme new 1593:1595 arg1 insights 1597:1604 new insights 1593:1604 new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily 1593:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 1 44 theme CaSR 218:221 arg1 domain 198:203 the dimeric extracellular domain 172:203 the dimeric extracellular domain (ECD) of the CaSR 172:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 44 theme CaSR 218:221 arg1 CaSR 218:221 the CaSR 214:221 the CaSR 214:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 44 theme CaSR 218:221 arg1 ECD 206:208 ECD 206:208 ECD 206:208 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 7 45 theme direct 1427:1432 arg1 interaction 1434:1444 a direct interaction 1425:1444 a direct interaction between the CaSR ECD and l-Phe 1425:1475 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 9 46 theme superfamily 1792:1802 arg1 C 1778:1778 family C 1771:1778 family C of the GPCR superfamily 1771:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 2 47 theme direct 315:320 arg1 interaction 322:332 direct interaction 315:332 direct interaction of Ca(2+) and Phe with the ECD of the receptor 315:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 3 48 theme cell 838:841 arg1 line 843:846 the host cell line 829:846 the host cell line employed for recombinant expression 829:882 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 4 49 theme ECD 921:923 arg1 structures 979:988 similar secondary structures 961:988 similar secondary structures 961:988 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 49 theme ECD 921:923 arg1 forms 903:907 Both glycosylated forms 885:907 Both glycosylated forms of the CaSR ECD 885:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 49 theme ECD 921:923 arg1 dimers 942:947 dimers 942:947 dimers 942:947 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 9 50 theme other 1754:1758 arg1 members 1760:1766 other members 1754:1766 other members of family C of the GPCR superfamily 1754:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 2 51 theme ECD 505:507 arg1 degree 519:524 its high degree 510:524 its high degree of glycosylation 510:541 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 51 theme ECD 505:507 arg1 lack 552:555 the lack 548:555 the lack of biophysical methods to monitor weak interactions in solution 548:619 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 51 theme ECD 505:507 arg1 size 493:496 the large size 483:496 the large size of the ECD 483:507 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 8 52 theme ECD 1556:1558 arg1 affinity 1535:1542 the binding affinity 1523:1542 the binding affinity of the CaSR ECD for Ca(2+) 1523:1569 We further demonstrated that l-Phe increases the binding affinity of the CaSR ECD for Ca(2+). 25305020 3 53 theme host 833:836 arg1 line 843:846 the host cell line 829:846 the host cell line employed for recombinant expression 829:882 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 54 theme recombinant 861:871 arg1 expression 873:882 recombinant expression 861:882 recombinant expression 861:882 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 4 55 theme 50 997:998 arg1 % 999:999 % 999:999 % 999:999 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 1 56 theme dimeric 176:182 arg1 domain 198:203 the dimeric extracellular domain 172:203 the dimeric extracellular domain (ECD) of the CaSR 172:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 56 theme dimeric 176:182 arg1 CaSR 218:221 the CaSR 214:221 the CaSR 214:221 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 1 56 theme dimeric 176:182 arg1 ECD 206:208 ECD 206:208 ECD 206:208 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 4 57 theme β-sheet 1016:1022 arg1 content 1024:1030 ∼ 20% β-sheet content 1010:1030 ∼ 20% β-sheet content 1010:1030 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 7 58 theme first 1414:1418 arg1 time 1420:1423 the first time 1410:1423 the first time 1410:1423 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 4 59 theme secondary 969:977 arg1 structures 979:988 similar secondary structures 961:988 similar secondary structures 961:988 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 59 theme secondary 969:977 arg1 dimers 942:947 dimers 942:947 dimers 942:947 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 59 theme secondary 969:977 arg1 forms 903:907 Both glycosylated forms 885:907 Both glycosylated forms of the CaSR ECD 885:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 2 60 from impact 399:404 arg1 changes 453:459 associated conformational changes 427:459 associated conformational changes 427:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 60 from impact 399:404 arg1 structure 413:421 its structure 409:421 its structure 409:421 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 9 61 theme amino 1646:1650 arg1 acids 1652:1656 amino acids 1646:1656 amino acids 1646:1656 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 3 62 theme present 629:635 arg1 study 637:641 the present study 625:641 the present study 625:641 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 63 theme glycosylated 660:671 arg1 domain 687:692 the glycosylated extracellular domain 656:692 the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 656:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 63 theme glycosylated 660:671 arg1 receptor 713:720 calcium-sensing receptor 697:720 calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 697:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 1 64 theme in 125:126 arg1 studies 144:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 7 65 theme difference 1367:1376 arg1 approaches 1382:1391 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 2 66 theme large 487:491 arg1 size 493:496 the large size 483:496 the large size of the ECD 483:507 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 6 67 theme ECD 1342:1344 arg1 forms 1329:1333 specific (15)N Phe-labeled forms 1302:1333 specific (15)N Phe-labeled forms of the ECD 1302:1344 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 1 68 theme functional 133:142 arg1 studies 144:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies 116:150 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 7 69 theme Saturation 1347:1356 arg1 difference 1367:1376 Saturation transfer difference 1347:1376 Saturation transfer difference NMR approaches 1347:1391 Saturation transfer difference NMR approaches demonstrated for the first time a direct interaction between the CaSR ECD and l-Phe. 25305020 3 70 theme calcium-sensing 697:711 arg1 residues 736:743 residues 20-612 736:750 residues 20-612 736:750 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 70 theme calcium-sensing 697:711 arg1 CaSR 723:726 CaSR 723:726 CaSR 723:726 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 70 theme calcium-sensing 697:711 arg1 ECD 730:732 ECD 730:732 ECD 730:732 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 70 theme calcium-sensing 697:711 arg1 receptor 713:720 calcium-sensing receptor 697:720 calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 697:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 6 71 theme local 1187:1191 arg1 changes 1208:1214 The local conformational changes 1183:1214 The local conformational changes of the proteins induced by their interactions with Ca(2+) 1183:1272 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 3 72 gly glycosylated 660:671 arg1 domain 687:692 the glycosylated extracellular domain 656:692 the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 656:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 3 72 gly glycosylated 660:671 arg1 receptor 713:720 calcium-sensing receptor 697:720 calcium-sensing receptor (CaSR) (ECD) (residues 20-612) 697:751 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 5 73 theme spectroscopic 1082:1094 arg1 methods 1096:1102 various spectroscopic methods 1074:1102 various spectroscopic methods 1074:1102 Using various spectroscopic methods, we have shown that both protein variants bind Ca(2+) with a Kd of 3.0-5.0 mm. 25305020 6 74 theme Phe-labeled 1317:1327 arg1 forms 1329:1333 specific (15)N Phe-labeled forms 1302:1333 specific (15)N Phe-labeled forms of the ECD 1302:1344 The local conformational changes of the proteins induced by their interactions with Ca(2+) were visualized by NMR with specific (15)N Phe-labeled forms of the ECD. 25305020 2 75 theme resultant 389:397 arg1 impact 399:404 the resultant impact 385:404 the resultant impact on its structure and associated conformational changes 385:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 0 76 theme receptor 106:113 arg1 receptor 106:113 the calcium-sensing receptor 86:113 the calcium-sensing receptor 86:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 0 76 theme receptor 106:113 arg1 domain 76:81 the extracellular domain 58:81 the extracellular domain of the calcium-sensing receptor 58:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 2 77 theme associated 427:436 arg1 changes 453:459 associated conformational changes 427:459 associated conformational changes 427:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 9 78 theme structural 1720:1729 arg1 properties 1731:1740 the structural properties 1716:1740 the structural properties of CaSR and other members of family C of the GPCR superfamily 1716:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 2 79 theme methods 572:578 arg1 degree 519:524 its high degree 510:524 its high degree of glycosylation 510:541 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 79 theme methods 572:578 arg1 lack 552:555 the lack 548:555 the lack of biophysical methods to monitor weak interactions in solution 548:619 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 79 theme methods 572:578 arg1 size 493:496 the large size 483:496 the large size of the ECD 483:507 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 80 theme Ca 337:338 arg1 interaction 322:332 direct interaction 315:332 direct interaction of Ca(2+) and Phe with the ECD of the receptor 315:379 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 80 theme Ca 337:338 arg1 impact 399:404 the resultant impact 385:404 the resultant impact on its structure and associated conformational changes 385:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 4 81 theme glycosylated 890:901 arg1 structures 979:988 similar secondary structures 961:988 similar secondary structures 961:988 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 81 theme glycosylated 890:901 arg1 forms 903:907 Both glycosylated forms 885:907 Both glycosylated forms of the CaSR ECD 885:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 81 theme glycosylated 890:901 arg1 dimers 942:947 dimers 942:947 dimers 942:947 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 82 theme Trp 1051:1053 arg1 environment 1055:1065 a well buried Trp environment 1037:1065 a well buried Trp environment 1037:1065 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 5 83 theme protein 1129:1135 arg1 variants 1137:1144 both protein variants 1124:1144 both protein variants 1124:1144 Using various spectroscopic methods, we have shown that both protein variants bind Ca(2+) with a Kd of 3.0-5.0 mm. 25305020 0 84 theme multiple 24:31 arg1 interactions 40:51 multiple ligand interactions 24:51 multiple ligand interactions with the extracellular domain of the calcium-sensing receptor 24:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 9 85 theme properties 1731:1740 arg1 exploration 1701:1711 exploration 1701:1711 exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily 1701:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 9 86 theme members 1760:1766 arg1 properties 1731:1740 the structural properties 1716:1740 the structural properties of CaSR and other members of family C of the GPCR superfamily 1716:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 0 87 theme interactions 40:51 arg1 determination 7:19 Direct determination 0:19 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor 0:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 4 88 gly glycosylated 890:901 arg1 structures 979:988 similar secondary structures 961:988 similar secondary structures 961:988 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 88 gly glycosylated 890:901 arg1 forms 903:907 Both glycosylated forms 885:907 Both glycosylated forms of the CaSR ECD 885:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 88 gly glycosylated 890:901 arg1 ECD 921:923 the CaSR ECD 912:923 the CaSR ECD 912:923 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 4 88 gly glycosylated 890:901 arg1 dimers 942:947 dimers 942:947 dimers 942:947 Both glycosylated forms of the CaSR ECD were purified as dimers and exhibit similar secondary structures with ∼ 50% α-helix, ∼ 20% β-sheet content, and a well buried Trp environment. 25305020 3 89 theme mannose 788:794 arg1 structures 805:814 either complex or high mannose N-glycan structures 765:814 either complex or high mannose N-glycan structures 765:814 In the present study, we purified the glycosylated extracellular domain of calcium-sensing receptor (CaSR) (ECD) (residues 20-612), containing either complex or high mannose N-glycan structures depending on the host cell line employed for recombinant expression. 25305020 9 90 theme family 1771:1776 arg1 C 1778:1778 family C 1771:1778 family C of the GPCR superfamily 1771:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 5 91 theme mm 1179:1180 arg1 Kd 1165:1166 a Kd 1163:1166 a Kd of 3.0-5.0 mm 1163:1180 Using various spectroscopic methods, we have shown that both protein variants bind Ca(2+) with a Kd of 3.0-5.0 mm. 25305020 2 92 from interaction 322:332 arg1 changes 453:459 associated conformational changes 427:459 associated conformational changes 427:459 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 2 92 from interaction 322:332 arg1 structure 413:421 its structure 409:421 its structure 409:421 However, direct interaction of Ca(2+) and Phe with the ECD of the receptor and the resultant impact on its structure and associated conformational changes have been hampered by the large size of the ECD, its high degree of glycosylation, and the lack of biophysical methods to monitor weak interactions in solution. 25305020 9 93 theme GPCR 1787:1790 arg1 superfamily 1792:1802 the GPCR superfamily 1783:1802 the GPCR superfamily 1783:1802 Our findings provide new insights into the mechanisms by which Ca(2+) and amino acids regulate the CaSR and may pave the way for exploration of the structural properties of CaSR and other members of family C of the GPCR superfamily. 25305020 1 94 theme crucial 231:237 arg1 role 239:242 a crucial role 229:242 a crucial role 229:242 Numerous in vivo functional studies have indicated that the dimeric extracellular domain (ECD) of the CaSR plays a crucial role in regulating Ca(2+) homeostasis by sensing Ca(2+) and l-Phe. 25305020 0 95 with interactions 40:51 arg1 receptor 106:113 the calcium-sensing receptor 86:113 the calcium-sensing receptor 86:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 0 95 with interactions 40:51 arg1 domain 76:81 the extracellular domain 58:81 the extracellular domain of the calcium-sensing receptor 58:113 Direct determination of multiple ligand interactions with the extracellular domain of the calcium-sensing receptor. 25305020 5 96 theme various 1074:1080 arg1 methods 1096:1102 various spectroscopic methods 1074:1102 various spectroscopic methods 1074:1102 Using various spectroscopic methods, we have shown that both protein variants bind Ca(2+) with a Kd of 3.0-5.0 mm. 26722726 4 0 theme protein 724:730 arg1 surface 732:738 the protein surface 720:738 the protein surface 720:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 6 1 theme interactions 985:996 arg1 probability 948:958 the probability 944:958 the probability of long- and short-range interactions forming differs between the folded and unfolded states 944:1051 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 2 2 dep in 291:292 arg1 vitro 294:298 vitro 294:298 vitro 294:298 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 6 3 theme short-range 973:983 arg1 interactions 985:996 long- and short-range interactions 963:996 long- and short-range interactions forming differs between the folded and unfolded states 963:1051 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 5 4 theme new 770:772 arg1 interactions 801:812 new short-range glycan-protein interactions 770:812 new short-range glycan-protein interactions that stabilize the conjugated protein 770:850 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 2 5 theme in 291:292 arg1 studies 314:320 in vitro and in silico studies 291:320 in vitro and in silico studies 291:320 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 4 6 theme destabilization 605:619 arg1 distortion 641:650 a conformational distortion 624:650 a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface 624:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 4 6 theme destabilization 605:619 arg1 origin 591:596 the origin 587:596 the origin of the destabilization 587:619 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 5 7 theme short-range 774:784 arg1 interactions 801:812 new short-range glycan-protein interactions 770:812 new short-range glycan-protein interactions that stabilize the conjugated protein 770:850 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 6 8 theme long- 963:967 arg1 interactions 985:996 long- and short-range interactions 963:996 long- and short-range interactions forming differs between the folded and unfolded states 963:1051 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 1 9 theme biophysical 169:179 arg1 properties 181:190 the biophysical properties 165:190 the biophysical properties of the modified protein 165:214 Glycosylation plays not only a functional role but can also modify the biophysical properties of the modified protein. 26722726 5 10 theme glycan-protein 786:799 arg1 interactions 801:812 new short-range glycan-protein interactions 770:812 new short-range glycan-protein interactions that stabilize the conjugated protein 770:850 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 6 11 theme unfolded 1037:1044 arg1 states 1046:1051 the folded and unfolded states 1022:1051 the folded and unfolded states 1022:1051 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 3 12 theme molecular 445:453 arg1 simulations 464:474 coarse-grained and all-atom molecular dynamics simulations 417:474 coarse-grained and all-atom molecular dynamics simulations 417:474 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 5 13 theme long-range 863:872 arg1 interactions 890:901 long-range protein-protein interactions 863:901 long-range protein-protein interactions 863:901 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 7 14 theme long-range 1168:1177 arg1 interactions 1179:1190 the short- and long-range interactions 1153:1190 the short- and long-range interactions 1153:1190 The destabilization originates not from simple loss of interactions but due to a trade-off between the short- and long-range interactions. 26722726 3 15 theme dynamics 455:462 arg1 simulations 464:474 coarse-grained and all-atom molecular dynamics simulations 417:474 coarse-grained and all-atom molecular dynamics simulations 417:474 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 0 16 theme Protein 25:31 arg1 Stability 47:55 Protein Thermodynamic Stability 25:55 Protein Thermodynamic Stability 25:55 Glycosylation May Reduce Protein Thermodynamic Stability by Inducing a Conformational Distortion. 26722726 2 17 theme in 304:305 arg1 studies 314:320 in vitro and in silico studies 291:320 in vitro and in silico studies 291:320 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 1 18 theme modified 199:206 arg1 protein 208:214 the modified protein 195:214 the modified protein 195:214 Glycosylation plays not only a functional role but can also modify the biophysical properties of the modified protein. 26722726 7 19 theme short- 1157:1162 arg1 interactions 1179:1190 the short- and long-range interactions 1153:1190 the short- and long-range interactions 1153:1190 The destabilization originates not from simple loss of interactions but due to a trade-off between the short- and long-range interactions. 26722726 1 20 theme protein 208:214 arg1 properties 181:190 the biophysical properties 165:190 the biophysical properties of the modified protein 165:214 Glycosylation plays not only a functional role but can also modify the biophysical properties of the modified protein. 26722726 5 21 theme conjugated 833:842 arg1 protein 844:850 the conjugated protein 829:850 the conjugated protein 829:850 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 3 22 theme stability 527:535 arg1 loss 519:522 the loss 515:522 the loss of stability of the MM1 protein by glycosylation 515:571 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 3 23 theme all-atom 436:443 arg1 simulations 464:474 coarse-grained and all-atom molecular dynamics simulations 417:474 coarse-grained and all-atom molecular dynamics simulations 417:474 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 0 24 theme Thermodynamic 33:45 arg1 Stability 47:55 Protein Thermodynamic Stability 25:55 Protein Thermodynamic Stability 25:55 Glycosylation May Reduce Protein Thermodynamic Stability by Inducing a Conformational Distortion. 26722726 4 25 with interaction 681:691 arg1 surface 732:738 the protein surface 720:738 the protein surface 720:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 5 26 theme protein-protein 874:888 arg1 interactions 890:901 long-range protein-protein interactions 863:901 long-range protein-protein interactions 863:901 Though glycosylation creates new short-range glycan-protein interactions that stabilize the conjugated protein, it breaks long-range protein-protein interactions. 26722726 6 27 theme folded 1026:1031 arg1 states 1046:1051 the folded and unfolded states 1022:1051 the folded and unfolded states 1022:1051 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 6 28 theme destabilizing 915:927 arg1 effect 929:934 a destabilizing effect 913:934 a destabilizing effect 913:934 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 2 29 theme natural 226:232 arg1 glycosylation 234:246 natural glycosylation 226:246 natural glycosylation 226:246 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 7 30 theme interactions 1109:1120 arg1 loss 1101:1104 simple loss 1094:1104 simple loss of interactions 1094:1120 The destabilization originates not from simple loss of interactions but due to a trade-off between the short- and long-range interactions. 26722726 2 31 dep in 304:305 arg1 silico 307:312 silico 307:312 silico 307:312 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 3 32 theme coarse-grained 417:430 arg1 simulations 464:474 coarse-grained and all-atom molecular dynamics simulations 417:474 coarse-grained and all-atom molecular dynamics simulations 417:474 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 7 33 theme simple 1094:1099 arg1 loss 1101:1104 simple loss 1094:1104 simple loss of interactions 1094:1120 The destabilization originates not from simple loss of interactions but due to a trade-off between the short- and long-range interactions. 26722726 1 34 theme functional 129:138 arg1 role 140:143 a functional role 127:143 a functional role 127:143 Glycosylation plays not only a functional role but can also modify the biophysical properties of the modified protein. 26722726 4 35 theme monosaccharide 700:713 arg1 interaction 681:691 the interaction 677:691 the interaction of the monosaccharide with the protein surface 677:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 4 36 theme conformational 626:639 arg1 distortion 641:650 a conformational distortion 624:650 a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface 624:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 4 36 theme conformational 626:639 arg1 origin 591:596 the origin 587:596 the origin of the destabilization 587:619 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 2 37 theme thermodynamic 369:381 arg1 destabilization 383:397 thermodynamic destabilization 369:397 thermodynamic destabilization 369:397 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 6 38 contain has 909:911 arg2 effect 929:934 a destabilizing effect 913:934 a destabilizing effect 913:934 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 6 38 contain has 909:911 arg1 This 904:907 This 904:907 This 904:907 This has a destabilizing effect because the probability of long- and short-range interactions forming differs between the folded and unfolded states. 26722726 0 39 theme Conformational 71:84 arg1 Distortion 86:95 a Conformational Distortion 69:95 a Conformational Distortion 69:95 Glycosylation May Reduce Protein Thermodynamic Stability by Inducing a Conformational Distortion. 26722726 2 40 theme protein 259:265 arg1 stabilization 267:279 protein stabilization 259:279 protein stabilization 259:279 Usually, natural glycosylation results in protein stabilization; however, in vitro and in silico studies showed that sometimes glycosylation results in thermodynamic destabilization. 26722726 4 41 theme protein 659:665 arg1 distortion 641:650 a conformational distortion 624:650 a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface 624:738 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 4 41 theme protein 659:665 arg1 origin 591:596 the origin 587:596 the origin of the destabilization 587:619 We show that the origin of the destabilization is a conformational distortion of the protein caused by the interaction of the monosaccharide with the protein surface. 26722726 3 42 theme MM1 544:546 arg1 protein 548:554 the MM1 protein 540:554 the MM1 protein 540:554 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26722726 3 43 theme protein 548:554 arg1 stability 527:535 stability 527:535 stability of the MM1 protein 527:554 Here, we applied coarse-grained and all-atom molecular dynamics simulations to understand the mechanism underlying the loss of stability of the MM1 protein by glycosylation. 26002521 0 0 theme Caenorhabditis 70:83 arg1 elegans 85:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 0 1 theme Mutant 63:68 arg1 elegans 85:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 1 2 theme simple 313:318 arg1 capacity 329:336 a rather simple glycomic capacity 304:336 a rather simple glycomic capacity 304:336 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 5 3 from NMR 1013:1015 arg1 conjunction 951:961 conjunction 951:961 conjunction with chemical and enzymatic treatments 951:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 6 4 theme " 1257:1257 arg1 β-galactose 1259:1269 a "bisecting" β-galactose 1245:1269 a "bisecting" β-galactose 1245:1269 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 1 5 theme glycomic 320:327 arg1 capacity 329:336 a rather simple glycomic capacity 304:336 a rather simple glycomic capacity 304:336 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 4 6 theme nematode 797:804 arg1 strain 806:811 a nematode strain 795:811 a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans 795:912 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 3 7 theme residues 496:503 arg1 linkage 472:478 the linkage 468:478 the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core 468:540 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 8 8 theme modifications 1450:1462 arg1 presence 1423:1430 the presence 1419:1430 the presence of novel N-glycan modifications 1419:1462 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 0 9 theme elegans 85:91 arg1 N-Glycans 36:44 N-Glycans 36:44 N-Glycans of Wild-type and Mutant Caenorhabditis elegans 36:91 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 6 10 contain carries 1171:1177 arg1 trisubstituted 1149:1162 trisubstituted 1149:1162 trisubstituted 1149:1162 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 10 contain carries 1171:1177 arg1 β-mannose 1132:1140 the core β-mannose 1123:1140 the core β-mannose 1123:1140 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 10 contain carries 1171:1177 arg2 β-galactose 1259:1269 a "bisecting" β-galactose 1245:1269 a "bisecting" β-galactose 1245:1269 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 10 contain carries 1171:1177 arg2 residues 1226:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 1 11 theme existing 247:254 arg1 impression 256:265 the existing impression 243:265 the existing impression that "simple" organisms possess also a rather simple glycomic capacity 243:336 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 1 12 gly N-glycosylation 98:112 arg1 elegans 151:157 the model nematode Caenorhabditis elegans 117:157 the model nematode Caenorhabditis elegans 117:157 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 4 13 theme responsible 718:728 arg1 genes 712:716 the three genes 702:716 the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8) 702:775 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 2 14 theme fucose 414:419 arg1 residues 421:428 up to four fucose residues 403:428 up to four fucose residues 403:428 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 2 15 with N-glycans 388:396 arg1 residues 421:428 up to four fucose residues 403:428 up to four fucose residues 403:428 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 3 16 theme fucose 489:494 arg1 residues 496:503 three fucose residues 483:503 three fucose residues 483:503 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 5 17 with conjunction 951:961 arg1 treatments 991:1000 chemical and enzymatic treatments 968:1000 chemical and enzymatic treatments 968:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 4 18 dep fucosylation 739:750 arg1 fut-8 770:774 fut-8 770:774 fut-8 770:774 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 4 18 dep fucosylation 739:750 arg1 fut-6 760:764 fut-6 760:764 fut-6 760:764 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 4 18 dep fucosylation 739:750 arg1 fut-1 753:757 fut-1 753:757 fut-1 753:757 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 8 19 theme biological 1517:1526 arg1 functions 1528:1536 the biological functions 1513:1536 the biological functions 1513:1536 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 4 20 dep strain 806:811 arg1 retaining 858:866 retaining 858:866 still retaining maximally one fucose residue on its N-glycans 852:912 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 4 20 dep strain 806:811 arg1 lacking 813:819 lacking 813:819 lacking products of these enzymes 813:845 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 2 21 dep four 409:412 arg1 to 406:407 to 406:407 to 406:407 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 4 22 from deletions 689:697 arg1 genes 712:716 the three genes 702:716 the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8) 702:775 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 8 23 theme N-glycan 1441:1448 arg1 modifications 1450:1462 novel N-glycan modifications 1435:1462 novel N-glycan modifications 1435:1462 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 1 24 contain possess 291:297 arg1 organisms 281:289 "simple" organisms 272:289 "simple" organisms 272:289 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 1 24 contain possess 291:297 arg2 capacity 329:336 a rather simple glycomic capacity 304:336 a rather simple glycomic capacity 304:336 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 1 24 contain possess 291:297 arg2 impression 256:265 the existing impression 243:265 the existing impression that "simple" organisms possess also a rather simple glycomic capacity 243:336 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 5 25 from HPLC 943:946 arg1 conjunction 951:961 conjunction 951:961 conjunction with chemical and enzymatic treatments 951:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 8 26 theme novel 1435:1439 arg1 modifications 1450:1462 novel N-glycan modifications 1435:1462 novel N-glycan modifications 1435:1462 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 4 27 theme enzymes 839:845 arg1 products 821:828 products 821:828 products of these enzymes 821:845 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 3 28 theme N'-diacetylchitobiosyl 514:535 arg1 N 512:512 the N 508:512 the N 508:512 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 3 28 theme N'-diacetylchitobiosyl 514:535 arg1 core 537:540 N'-diacetylchitobiosyl core 514:540 N'-diacetylchitobiosyl core 514:540 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 6 29 theme glycomic 1086:1093 arg1 subpool 1095:1101 this glycomic subpool 1081:1101 this glycomic subpool 1081:1101 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 5 30 theme enzymatic 981:989 arg1 treatments 991:1000 chemical and enzymatic treatments 968:1000 chemical and enzymatic treatments 968:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 5 31 from spectrometry 926:937 arg1 conjunction 951:961 conjunction 951:961 conjunction with chemical and enzymatic treatments 951:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 1 32 theme Caenorhabditis 136:149 arg1 elegans 151:157 the model nematode Caenorhabditis elegans 117:157 the model nematode Caenorhabditis elegans 117:157 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 1 33 theme simple 273:278 arg1 organisms 281:289 "simple" organisms 272:289 "simple" organisms 272:289 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 5 34 theme mass 921:924 arg1 spectrometry 926:937 mass spectrometry 921:937 mass spectrometry 921:937 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 2 35 theme laboratories 374:385 arg1 number 364:369 a number 362:369 a number of laboratories 362:385 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 1 36 theme " 279:279 arg1 organisms 281:289 "simple" organisms 272:289 "simple" organisms 272:289 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 6 37 mod modified 1303:1310 arg3 fucose 1317:1322 fucose 1317:1322 fucose 1317:1322 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 37 mod modified 1303:1310 arg1 β-galactose 1259:1269 a "bisecting" β-galactose 1245:1269 a "bisecting" β-galactose 1245:1269 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 37 mod modified 1303:1310 arg3 methylfucose 1327:1338 methylfucose 1327:1338 methylfucose 1327:1338 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 5 38 theme N-glycans 1063:1071 arg1 set 1032:1034 a set 1030:1034 a set of α-mannosidase-resistant N-glycans 1030:1071 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 5 38 theme N-glycans 1063:1071 arg1 N-glycans 1063:1071 α-mannosidase-resistant N-glycans 1039:1071 α-mannosidase-resistant N-glycans 1039:1071 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 6 39 theme α1,6-mannose 1213:1224 arg1 residues 1226:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 4 40 theme fucose 882:887 arg1 residue 889:895 maximally one fucose residue 868:895 maximally one fucose residue on its N-glycans 868:912 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 6 41 theme core 1127:1130 arg1 β-mannose 1132:1140 the core β-mannose 1123:1140 the core β-mannose 1123:1140 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 6 41 theme core 1127:1130 arg1 trisubstituted 1149:1162 trisubstituted 1149:1162 trisubstituted 1149:1162 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 2 42 from studies 351:357 arg1 number 364:369 a number 362:369 a number of laboratories 362:385 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 0 43 theme N-Glycans 36:44 arg1 Galactose 10:18 Galactose 10:18 Galactose 10:18 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 0 43 theme N-Glycans 36:44 arg1 Feature 25:31 a Feature 23:31 a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans 23:91 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 8 44 theme targeted 1482:1489 arg1 studies 1491:1497 more targeted studies 1477:1497 more targeted studies 1477:1497 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 3 45 theme fourth 614:619 arg1 fucose 621:626 the fourth fucose 610:626 the fourth fucose 610:626 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 5 46 theme α-mannosidase-resistant 1039:1061 arg1 N-glycans 1063:1071 α-mannosidase-resistant N-glycans 1039:1071 α-mannosidase-resistant N-glycans 1039:1071 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 8 47 theme nematode 1558:1565 arg1 glycans 1567:1573 nematode glycans 1558:1573 nematode glycans 1558:1573 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 4 48 with mutant 677:682 arg1 deletions 689:697 deletions 689:697 deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8) 689:775 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 6 49 theme α1,3- 1203:1207 arg1 residues 1226:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 3 50 theme fucose 621:626 arg1 nature 600:605 the nature 596:605 the nature of the fourth fucose 596:626 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 4 51 theme core 734:737 arg1 fucosylation 739:750 core fucosylation 734:750 core fucosylation (fut-1, fut-6 and fut-8) 734:775 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 1 52 theme model 121:125 arg1 elegans 151:157 the model nematode Caenorhabditis elegans 117:157 the model nematode Caenorhabditis elegans 117:157 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 6 53 theme ubiquitous 1192:1201 arg1 residues 1226:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 the ubiquitous α1,3- and α1,6-mannose residues 1188:1233 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 26002521 5 54 theme chemical 968:975 arg1 treatments 991:1000 chemical and enzymatic treatments 968:1000 chemical and enzymatic treatments 968:1000 Using mass spectrometry and HPLC in conjunction with chemical and enzymatic treatments as well as NMR, we examined a set of α-mannosidase-resistant N-glycans. 26002521 3 55 theme enzymatic 576:584 arg1 analyses 586:593 structural and enzymatic analyses 561:593 structural and enzymatic analyses 561:593 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 1 56 theme nematode 127:134 arg1 elegans 151:157 the model nematode Caenorhabditis elegans 117:157 the model nematode Caenorhabditis elegans 117:157 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 4 57 theme triple 670:675 arg1 mutant 677:682 a triple mutant 668:682 a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8) 668:775 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 2 58 theme previous 342:349 arg1 studies 351:357 previous studies 342:357 previous studies in a number of laboratories 342:385 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 8 59 theme glycans 1567:1573 arg1 functions 1528:1536 the biological functions 1513:1536 the biological functions 1513:1536 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 8 59 theme glycans 1567:1573 arg1 interactions 1542:1553 interactions 1542:1553 interactions of nematode glycans 1542:1573 Our data, showing the presence of novel N-glycan modifications, will enable more targeted studies to understand the biological functions and interactions of nematode glycans. 26002521 4 60 from residue 889:895 arg1 N-glycans 904:912 its N-glycans 900:912 its N-glycans 900:912 By constructing a triple mutant with deletions in the three genes responsible for core fucosylation (fut-1, fut-6 and fut-8), we have produced a nematode strain lacking products of these enzymes, but still retaining maximally one fucose residue on its N-glycans. 26002521 0 61 theme Wild-type 49:57 arg1 elegans 85:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Wild-type and Mutant Caenorhabditis elegans 49:91 Bisecting Galactose as a Feature of N-Glycans of Wild-type and Mutant Caenorhabditis elegans. 26002521 3 62 theme structural 561:570 arg1 analyses 586:593 structural and enzymatic analyses 561:593 structural and enzymatic analyses 561:593 However, although the linkage of three fucose residues to the N,N'-diacetylchitobiosyl core has been proven by structural and enzymatic analyses, the nature of the fourth fucose has remained uncertain. 26002521 2 63 located detected 440:447 arg2 N-glycans 388:396 N-glycans 388:396 N-glycans with up to four fucose residues 388:428 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 2 63 located detected 440:447 arg1 studies 351:357 previous studies 342:357 previous studies in a number of laboratories 342:385 In previous studies in a number of laboratories, N-glycans with up to four fucose residues have been detected. 26002521 1 64 theme elegans 151:157 arg1 variable 183:190 variable 183:190 variable 183:190 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 1 64 theme elegans 151:157 arg1 N-glycosylation 98:112 The N-glycosylation 94:112 The N-glycosylation of the model nematode Caenorhabditis elegans 94:157 The N-glycosylation of the model nematode Caenorhabditis elegans has proven to be highly variable and rather complex; it is an example to contradict the existing impression that "simple" organisms possess also a rather simple glycomic capacity. 26002521 6 65 theme bisecting 1248:1256 arg1 β-galactose 1259:1269 a "bisecting" β-galactose 1245:1269 a "bisecting" β-galactose 1245:1269 Within this glycomic subpool, we can reveal that the core β-mannose can be trisubstituted and so carries not only the ubiquitous α1,3- and α1,6-mannose residues, but also a "bisecting" β-galactose, which is substoichiometrically modified with fucose or methylfucose. 27882844 4 0 theme temperature 801:811 arg1 irrespective 781:792 irrespective 781:792 irrespective 781:792 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 6 1 theme native 1237:1242 arg1 form 1261:1264 native and glycosylated form 1237:1264 native and glycosylated form 1237:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 4 2 from rigid 877:881 arg1 form 956:959 its native and glycosylated form 928:959 its native and glycosylated form 928:959 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 3 theme ice-binding 817:827 arg1 IBS 838:840 IBS 838:840 IBS 838:840 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 3 theme ice-binding 817:827 arg1 rigid 877:881 rigid 877:881 rigid 877:881 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 3 theme ice-binding 817:827 arg1 surface 829:835 the ice-binding surface 813:835 irrespective of the temperature the ice-binding surface (IBS) of the protein 781:856 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 3 theme ice-binding 817:827 arg1 NonIBS 917:922 NonIBS 917:922 NonIBS 917:922 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 6 4 theme neighbor 1101:1108 arg1 orientation 1110:1120 nearest neighbor orientation 1093:1120 nearest neighbor orientation 1093:1120 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 3 5 theme protein 652:658 arg1 rigidity 605:612 the conformational rigidity 586:612 the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface 586:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 6 6 theme water 1192:1196 arg1 molecule 1198:1205 water molecule 1192:1205 water molecule near these binding surface in native and glycosylated form 1192:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 6 7 from density 1181:1187 arg1 form 1261:1264 native and glycosylated form 1237:1264 native and glycosylated form 1237:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 7 8 theme strong 1345:1350 arg1 peak 1352:1355 a strong peak 1343:1355 a strong peak than the native one 1343:1375 The glycosylated form shows a strong peak than the native one. 27882844 9 9 theme prism 1734:1738 arg1 plane 1740:1744 the prism plane 1730:1744 the prism plane of ice 1730:1751 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 1 10 contain have 238:241 arg1 AFPs 174:177 AFPs 174:177 AFPs 174:177 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 10 contain have 238:241 arg1 proteins 164:171 Antifreeze proteins 153:171 Antifreeze proteins (AFPs) 153:178 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 10 contain have 238:241 arg2 ability 247:253 the ability to permit their survival in subzero environments by thermal hysteresis mechanism 243:334 the ability to permit their survival in subzero environments by thermal hysteresis mechanism 243:334 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 6 11 theme distribution 1067:1078 arg1 function 1080:1087 density distribution function 1059:1087 density distribution function 1059:1087 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 8 12 from increase 1491:1498 arg1 temperature 1503:1513 temperature 1503:1513 temperature 1503:1513 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 3 13 theme different 514:522 arg1 temperatures 524:535 two different temperatures 510:535 two different temperatures (277 and 298 K) 510:551 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 7 14 theme glycosylated 1319:1330 arg1 form 1332:1335 The glycosylated form 1315:1335 The glycosylated form 1315:1335 The glycosylated form shows a strong peak than the native one. 27882844 3 15 theme aqueous 663:669 arg1 medium 671:676 aqueous medium 663:676 aqueous medium 663:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 16 theme antifreeze 100:109 arg1 protein 111:117 the Ocean Pout type III antifreeze protein 76:117 the Ocean Pout type III antifreeze protein 76:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 8 17 theme water 1547:1551 arg1 oxygen 1553:1558 the water oxygen 1543:1558 the water oxygen 1543:1558 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 6 18 gly glycosylated 1248:1259 arg1 form 1261:1264 native and glycosylated form 1237:1264 native and glycosylated form 1237:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 4 19 dep rigid 877:881 arg1 rigid 877:881 rigid 877:881 rigid 877:881 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 19 dep rigid 877:881 arg1 surface 829:835 the ice-binding surface 813:835 irrespective of the temperature the ice-binding surface (IBS) of the protein 781:856 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 19 dep rigid 877:881 arg1 NonIBS 917:922 NonIBS 917:922 NonIBS 917:922 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 6 20 theme individual 1144:1153 arg1 surfaces 1159:1166 individual two surfaces 1144:1166 individual two surfaces 1144:1166 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 3 21 theme structural 686:695 arg1 arrangements 697:708 the structural arrangements 682:708 the structural arrangements of water molecules hydrating its ice-binding surface 682:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 22 theme molecular 122:130 arg1 simulation 141:150 molecular dynamics simulation 122:150 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 0:151 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 9 23 theme molecular 1664:1672 arg1 reason 1674:1679 a possible molecular reason 1653:1679 a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice 1653:1751 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 8 24 theme rotational 1383:1392 arg1 function 1411:1418 rotational auto correlation function 1383:1418 rotational auto correlation function of water molecules around ice-binding sites 1383:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 6 25 theme binding 1218:1224 arg1 surface 1226:1232 these binding surface 1212:1232 these binding surface 1212:1232 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 5 26 theme Hydrophilic 962:972 arg1 Q44 996:998 Q44 996:998 Q44 996:998 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 26 theme Hydrophilic 962:972 arg1 residues 974:981 Hydrophilic residues 962:981 Hydrophilic residues N14, T18 and Q44 962:998 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 26 theme Hydrophilic 962:972 arg1 T18 988:990 T18 988:990 T18 988:990 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 26 theme Hydrophilic 962:972 arg1 N14 983:985 N14 983:985 N14 983:985 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 4 27 theme non 892:894 arg1 surface 908:914 its non ice-binding surface 888:914 its non ice-binding surface 888:914 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 8 28 theme correlation 1399:1409 arg1 function 1411:1418 rotational auto correlation function 1383:1418 rotational auto correlation function of water molecules around ice-binding sites 1383:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 3 29 theme molecules 719:727 arg1 arrangements 697:708 the structural arrangements 682:708 the structural arrangements of water molecules hydrating its ice-binding surface 682:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 29 theme molecules 719:727 arg1 protein 652:658 the Ocean pout type III antifreeze protein 617:658 the Ocean pout type III antifreeze protein in aqueous medium 617:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 30 theme ice-binding 53:63 arg1 surface 65:71 the ice-binding surface 49:71 the ice-binding surface of the Ocean Pout type III antifreeze protein 49:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 5 31 theme antifreeze 1017:1026 arg1 activity 1028:1035 antifreeze activity 1017:1035 antifreeze activity 1017:1035 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 9 32 theme ice-binding 1692:1702 arg1 activity 1704:1711 the ice-binding activity 1688:1711 the ice-binding activity of ocean pout 1688:1725 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 6 33 theme nonbinding 1295:1304 arg1 surface 1306:1312 the nonbinding surface 1291:1312 the nonbinding surface 1291:1312 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 4 34 from form 956:959 arg1 rigid 877:881 rigid 877:881 rigid 877:881 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 34 from form 956:959 arg1 surface 829:835 the ice-binding surface 813:835 irrespective of the temperature the ice-binding surface (IBS) of the protein 781:856 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 34 from form 956:959 arg1 NonIBS 917:922 NonIBS 917:922 NonIBS 917:922 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 0 35 from Effect 0:5 arg1 behavior 37:44 hydration behavior 27:44 hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein 27:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 0 36 theme Ocean 80:84 arg1 protein 111:117 the Ocean Pout type III antifreeze protein 76:117 the Ocean Pout type III antifreeze protein 76:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 1 37 theme hysteresis 315:324 arg1 mechanism 326:334 thermal hysteresis mechanism 307:334 thermal hysteresis mechanism 307:334 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 9 38 theme pout 1722:1725 arg1 activity 1704:1711 the ice-binding activity 1688:1711 the ice-binding activity of ocean pout 1688:1725 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 8 39 theme ice-binding 1446:1456 arg1 sites 1458:1462 ice-binding sites 1446:1462 ice-binding sites 1446:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 0 40 theme type 91:94 arg1 protein 111:117 the Ocean Pout type III antifreeze protein 76:117 the Ocean Pout type III antifreeze protein 76:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 4 41 theme native 932:937 arg1 form 956:959 its native and glycosylated form 928:959 its native and glycosylated form 928:959 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 42 theme glycosylated 943:954 arg1 form 956:959 its native and glycosylated form 928:959 its native and glycosylated form 928:959 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 2 43 theme inhibition 380:389 arg1 mechanism 356:364 the exact mechanism 346:364 the exact mechanism of ice growth inhibition 346:389 However, the exact mechanism of ice growth inhibition is still not clearly understood. 27882844 0 44 theme glycosylation 10:22 arg1 Effect 0:5 Effect 0:5 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 0:151 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 9 45 theme ice 1749:1751 arg1 plane 1740:1744 the prism plane 1730:1744 the prism plane of ice 1730:1751 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 2 46 theme ice 369:371 arg1 inhibition 380:389 ice growth inhibition 369:389 ice growth inhibition 369:389 However, the exact mechanism of ice growth inhibition is still not clearly understood. 27882844 3 47 dep temperatures 524:535 arg1 277 538:540 277 538:540 277 538:540 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 47 dep temperatures 524:535 arg1 298 K 546:550 298 K 546:550 298 K 546:550 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 48 theme hydration 27:35 arg1 behavior 37:44 hydration behavior 27:44 hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein 27:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 3 49 theme explicit 440:447 arg1 MD 469:470 MD 469:470 MD 469:470 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 49 theme explicit 440:447 arg1 dynamics 459:466 explicit molecular dynamics 440:466 four long explicit molecular dynamics (MD) simulations 430:483 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 1 50 theme subzero 283:289 arg1 environments 291:302 subzero environments 283:302 subzero environments 283:302 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 4 51 gly glycosylated 943:954 arg1 form 956:959 its native and glycosylated form 928:959 its native and glycosylated form 928:959 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 8 52 theme hydrogen 1568:1575 arg1 bond 1577:1580 the hydrogen bond 1564:1580 the hydrogen bond acceptor group 1564:1595 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 6 53 from molecule 1198:1205 arg1 form 1261:1264 native and glycosylated form 1237:1264 native and glycosylated form 1237:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 8 54 theme strong 1516:1521 arg1 interaction 1523:1533 strong interaction 1516:1533 strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface 1516:1626 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 3 55 theme conformational 590:603 arg1 rigidity 605:612 the conformational rigidity 586:612 the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface 586:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 8 56 theme acceptor 1582:1589 arg1 group 1591:1595 the hydrogen bond acceptor group 1564:1595 the hydrogen bond acceptor group 1564:1595 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 0 57 dep Effect 0:5 arg1 simulation 141:150 molecular dynamics simulation 122:150 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 0:151 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 3 58 from rigidity 605:612 arg1 medium 671:676 aqueous medium 663:676 aqueous medium 663:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 59 theme type 632:635 arg1 protein 652:658 the Ocean pout type III antifreeze protein 617:658 the Ocean pout type III antifreeze protein in aqueous medium 617:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 1 60 from survival 271:278 arg1 environments 291:302 subzero environments 283:302 subzero environments 283:302 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 3 61 theme dynamics 459:466 arg1 simulations 473:483 four long explicit molecular dynamics (MD) simulations 430:483 four long explicit molecular dynamics (MD) simulations 430:483 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 7 62 gly glycosylated 1319:1330 arg1 form 1332:1335 The glycosylated form 1315:1335 The glycosylated form 1315:1335 The glycosylated form shows a strong peak than the native one. 27882844 5 63 dep residues 974:981 arg1 Q44 996:998 Q44 996:998 Q44 996:998 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 63 dep residues 974:981 arg1 residues 974:981 Hydrophilic residues 962:981 Hydrophilic residues N14, T18 and Q44 962:998 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 63 dep residues 974:981 arg1 T18 988:990 T18 988:990 T18 988:990 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 5 63 dep residues 974:981 arg1 N14 983:985 N14 983:985 N14 983:985 Hydrophilic residues N14, T18 and Q44 are essential to antifreeze activity. 27882844 3 64 theme Ocean 621:625 arg1 protein 652:658 the Ocean pout type III antifreeze protein 617:658 the Ocean pout type III antifreeze protein in aqueous medium 617:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 6 65 theme molecule 1198:1205 arg1 density 1181:1187 density 1181:1187 density of water molecule near these binding surface in native and glycosylated form 1181:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 8 66 theme protein-binding 1604:1618 arg1 surface 1620:1626 the protein-binding surface 1600:1626 the protein-binding surface 1600:1626 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 6 67 theme nearest 1093:1099 arg1 orientation 1110:1120 nearest neighbor orientation 1093:1120 nearest neighbor orientation 1093:1120 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 1 68 theme Antifreeze 153:162 arg1 AFPs 174:177 AFPs 174:177 AFPs 174:177 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 68 theme Antifreeze 153:162 arg1 proteins 164:171 Antifreeze proteins 153:171 Antifreeze proteins (AFPs) 153:178 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 7 69 theme native 1366:1371 arg1 one 1373:1375 the native one 1362:1375 the native one 1362:1375 The glycosylated form shows a strong peak than the native one. 27882844 6 70 theme function 1080:1087 arg1 plots 1122:1126 Radial distribution, density distribution function and nearest neighbor orientation plots 1038:1126 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces 1038:1166 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 3 71 theme antifreeze 641:650 arg1 protein 652:658 the Ocean pout type III antifreeze protein 617:658 the Ocean pout type III antifreeze protein in aqueous medium 617:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 4 72 theme irrespective 781:792 arg1 IBS 838:840 IBS 838:840 IBS 838:840 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 72 theme irrespective 781:792 arg1 rigid 877:881 rigid 877:881 rigid 877:881 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 72 theme irrespective 781:792 arg1 surface 829:835 the ice-binding surface 813:835 irrespective of the temperature the ice-binding surface (IBS) of the protein 781:856 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 72 theme irrespective 781:792 arg1 NonIBS 917:922 NonIBS 917:922 NonIBS 917:922 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 6 73 theme density 1059:1065 arg1 function 1080:1087 density distribution function 1059:1087 density distribution function 1059:1087 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 3 74 from protein 652:658 arg1 medium 671:676 aqueous medium 663:676 aqueous medium 663:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 75 theme protein 111:117 arg1 surface 65:71 the ice-binding surface 49:71 the ice-binding surface of the Ocean Pout type III antifreeze protein 49:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 6 76 theme distribution 1045:1056 arg1 plots 1122:1126 Radial distribution, density distribution function and nearest neighbor orientation plots 1038:1126 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces 1038:1166 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 6 77 theme glycosylated 1248:1259 arg1 form 1261:1264 native and glycosylated form 1237:1264 native and glycosylated form 1237:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 4 78 theme protein 850:856 arg1 IBS 838:840 IBS 838:840 IBS 838:840 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 78 theme protein 850:856 arg1 rigid 877:881 rigid 877:881 rigid 877:881 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 78 theme protein 850:856 arg1 surface 829:835 the ice-binding surface 813:835 irrespective of the temperature the ice-binding surface (IBS) of the protein 781:856 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 4 78 theme protein 850:856 arg1 NonIBS 917:922 NonIBS 917:922 NonIBS 917:922 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 0 79 from surface 65:71 arg1 behavior 37:44 hydration behavior 27:44 hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein 27:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 3 80 theme water 713:717 arg1 molecules 719:727 water molecules 713:727 water molecules hydrating its ice-binding surface 713:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 81 theme dynamics 132:139 arg1 simulation 141:150 molecular dynamics simulation 122:150 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 0:151 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 8 82 theme molecules 1429:1437 arg1 function 1411:1418 rotational auto correlation function 1383:1418 rotational auto correlation function of water molecules around ice-binding sites 1383:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 9 83 theme possible 1655:1662 arg1 reason 1674:1679 a possible molecular reason 1653:1679 a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice 1653:1751 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 8 84 theme auto 1394:1397 arg1 function 1411:1418 rotational auto correlation function 1383:1418 rotational auto correlation function of water molecules around ice-binding sites 1383:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 3 85 theme arrangements 697:708 arg1 rigidity 605:612 the conformational rigidity 586:612 the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface 586:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 86 theme ice-binding 743:753 arg1 surface 755:761 its ice-binding surface 739:761 its ice-binding surface 739:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 1 87 located found 181:185 arg1 vertebrates 198:208 certain vertebrates 190:208 certain vertebrates 190:208 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 87 located found 181:185 arg1 plants 211:216 plants 211:216 plants 211:216 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 87 located found 181:185 arg2 proteins 164:171 Antifreeze proteins 153:171 Antifreeze proteins (AFPs) 153:178 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 87 located found 181:185 arg2 AFPs 174:177 AFPs 174:177 AFPs 174:177 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 87 located found 181:185 arg1 fungi 219:223 fungi 219:223 fungi 219:223 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 1 87 located found 181:185 arg1 bacteria 229:236 bacteria 229:236 bacteria 229:236 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 8 88 theme water 1423:1427 arg1 molecules 1429:1437 water molecules 1423:1437 water molecules around ice-binding sites 1423:1462 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 6 89 theme orientation 1110:1120 arg1 plots 1122:1126 Radial distribution, density distribution function and nearest neighbor orientation plots 1038:1126 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces 1038:1166 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 1 90 theme certain 190:196 arg1 vertebrates 198:208 certain vertebrates 190:208 certain vertebrates 190:208 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 9 91 dep reason 1674:1679 arg1 behind 1681:1686 behind 1681:1686 behind the ice-binding activity of ocean pout at the prism plane of ice 1681:1751 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 4 92 theme ice-binding 896:906 arg1 surface 908:914 its non ice-binding surface 888:914 its non ice-binding surface 888:914 It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its non ice-binding surface (NonIBS) in its native and glycosylated form. 27882844 3 93 from arrangements 697:708 arg1 medium 671:676 aqueous medium 663:676 aqueous medium 663:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 0 94 theme Pout 86:89 arg1 protein 111:117 the Ocean Pout type III antifreeze protein 76:117 the Ocean Pout type III antifreeze protein 76:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 1 95 theme thermal 307:313 arg1 hysteresis 315:324 thermal hysteresis 307:324 thermal hysteresis mechanism 307:334 Antifreeze proteins (AFPs), found in certain vertebrates, plants, fungi and bacteria have the ability to permit their survival in subzero environments by thermal hysteresis mechanism. 27882844 9 96 theme ocean 1716:1720 arg1 pout 1722:1725 ocean pout 1716:1725 ocean pout 1716:1725 This provides a possible molecular reason behind the ice-binding activity of ocean pout at the prism plane of ice. 27882844 3 97 from medium 671:676 arg1 rigidity 605:612 the conformational rigidity 586:612 the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface 586:761 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 2 98 theme exact 350:354 arg1 mechanism 356:364 the exact mechanism 346:364 the exact mechanism of ice growth inhibition 346:389 However, the exact mechanism of ice growth inhibition is still not clearly understood. 27882844 8 99 from interaction 1523:1533 arg1 surface 1620:1626 the protein-binding surface 1600:1626 the protein-binding surface 1600:1626 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 2 100 theme growth 373:378 arg1 inhibition 380:389 ice growth inhibition 369:389 ice growth inhibition 369:389 However, the exact mechanism of ice growth inhibition is still not clearly understood. 27882844 6 101 from form 1261:1264 arg1 density 1181:1187 density 1181:1187 density of water molecule near these binding surface in native and glycosylated form 1181:1264 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 8 102 from function 1411:1418 arg1 prominent 1471:1479 prominent 1471:1479 prominent 1471:1479 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 0 103 gly glycosylation 10:22 arg1 hydration 27:35 hydration behavior 27:44 hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein 27:117 Effect of glycosylation on hydration behavior at the ice-binding surface of the Ocean Pout type III antifreeze protein: a molecular dynamics simulation. 27882844 6 104 theme Radial 1038:1043 arg1 distribution 1045:1056 Radial distribution 1038:1056 Radial distribution 1038:1056 Radial distribution, density distribution function and nearest neighbor orientation plots with respect to individual two surfaces confirm that density of water molecule near these binding surface in native and glycosylated form are relatively more than the nonbinding surface. 27882844 3 105 theme molecular 449:457 arg1 MD 469:470 MD 469:470 MD 469:470 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 105 theme molecular 449:457 arg1 dynamics 459:466 explicit molecular dynamics 440:466 four long explicit molecular dynamics (MD) simulations 430:483 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 8 106 theme bond 1577:1580 arg1 group 1591:1595 the hydrogen bond acceptor group 1564:1595 the hydrogen bond acceptor group 1564:1595 From rotational auto correlation function of water molecules around ice-binding sites, it is prominent that with increase in temperature, strong interaction between the water oxygen and the hydrogen bond acceptor group on the protein-binding surface decreases. 27882844 3 107 theme long 435:438 arg1 simulations 473:483 four long explicit molecular dynamics (MD) simulations 430:483 four long explicit molecular dynamics (MD) simulations 430:483 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 27882844 3 108 theme pout 627:630 arg1 protein 652:658 the Ocean pout type III antifreeze protein 617:658 the Ocean pout type III antifreeze protein in aqueous medium 617:676 Here, four long explicit molecular dynamics (MD) simulations have been carried out at two different temperatures (277 and 298 K) with and without glycan to study the conformational rigidity of the Ocean pout type III antifreeze protein in aqueous medium and the structural arrangements of water molecules hydrating its ice-binding surface. 28975713 0 0 theme Glycopeptide 70:81 arg1 Spectra 86:92 Intact Glycopeptide MS Spectra 63:92 Intact Glycopeptide MS Spectra 63:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 7 1 theme glycopeptide 1314:1325 arg1 matches 1332:1338 the most probable intact glycopeptide mass matches 1289:1338 the most probable intact glycopeptide mass matches 1289:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 7 2 theme CONCLUSIONS 1205:1215 arg1 PepSweetener 1240:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener 1205:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener 1205:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 6 3 theme fragmentation 1111:1123 arg1 diagram 1125:1131 an in silico peptide fragmentation diagram 1090:1131 an in silico peptide fragmentation diagram 1090:1131 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 3 4 theme correct 524:530 arg1 peptide 532:538 the correct peptide and glycan composition 520:561 peptide 532:538 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 9 5 theme resource 1523:1530 arg1 portal 1532:1537 Bioinformatics resource portal 1508:1537 Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/) 1508:1589 The tool is publicly hosted on ExPASy, the SIB Swiss Institute of Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/). 28975713 0 6 theme Intact 63:68 arg1 Spectra 86:92 Intact Glycopeptide MS Spectra 63:92 Intact Glycopeptide MS Spectra 63:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 5 7 from results 832:838 arg1 chart 867:871 an interactive heat-map chart 843:871 an interactive heat-map chart where theoretical glycopeptide tile colors 843:914 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 6 8 gly glycopeptide 1176:1187 arg2 glycopeptide 1176:1187 the correct glycopeptide identification 1164:1202 the correct glycopeptide identification 1164:1202 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 4 9 gly glycopeptide 626:637 arg2 glycopeptide 626:637 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 3 10 theme peptide 532:538 arg1 determination 503:515 the determination 499:515 the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass 499:607 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 9 11 theme Bioinformatics 1508:1521 arg1 portal 1532:1537 Bioinformatics resource portal 1508:1537 Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/) 1508:1589 The tool is publicly hosted on ExPASy, the SIB Swiss Institute of Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/). 28975713 0 12 theme Spectra 86:92 arg1 Annotation 49:58 Support Manual Annotation 34:58 Support Manual Annotation of Intact Glycopeptide MS Spectra 34:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 5 13 from mass 970:973 arg1 deviations 934:943 ppm deviations 930:943 ppm deviations from the query precursor mass 930:973 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 2 14 theme EXPERIMENTAL 287:298 arg1 DESIGN 300:305 EXPERIMENTAL DESIGN 287:305 EXPERIMENTAL DESIGN This exploratory tool 287:327 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 7 15 gly glycopeptide 1314:1325 arg2 glycopeptide 1314:1325 the most probable intact glycopeptide mass matches 1289:1338 the most probable intact glycopeptide mass matches 1289:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 4 16 theme theoretical 614:624 arg1 space 646:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 4 16 theme theoretical 614:624 arg1 customized 659:668 customized 659:668 customized 659:668 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 0 17 theme MS 83:84 arg1 Spectra 86:92 Intact Glycopeptide MS Spectra 63:92 Intact Glycopeptide MS Spectra 63:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 3 18 theme glycopeptide 568:579 arg1 peptide 532:538 the correct peptide and glycan composition 520:561 peptide 532:538 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 3 18 theme glycopeptide 568:579 arg1 composition 551:561 the correct peptide and glycan composition 520:561 composition 551:561 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 7 19 theme matches 1332:1338 arg1 selection 1276:1284 the selection 1272:1284 the selection of the most probable intact glycopeptide mass matches 1272:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 8 20 theme serum 1399:1403 arg1 samples 1413:1419 serum protein samples 1399:1419 serum protein samples 1399:1419 It is validated on serum protein samples and immunoglobulins. 28975713 3 21 gly glycopeptide 568:579 arg2 glycopeptide 568:579 a glycopeptide 566:579 a glycopeptide based on its precursor mass 566:607 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 2 22 theme error 435:439 arg1 range 441:445 the error range 431:445 the error range of the query precursor ion 431:472 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 3 23 theme composition 551:561 arg1 determination 503:515 the determination 499:515 the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass 499:607 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 4 24 from mode 691:694 arg1 space 646:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 4 24 from mode 691:694 arg1 customized 659:668 customized 659:668 customized 659:668 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 4 25 theme query 685:689 arg1 mode 691:694 an advanced query mode 673:694 an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters 673:795 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 2 26 theme ion 470:472 arg1 range 441:445 the error range 431:445 the error range of the query precursor ion 431:472 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 5 27 theme heat-map 858:865 arg1 chart 867:871 an interactive heat-map chart 843:871 an interactive heat-map chart where theoretical glycopeptide tile colors 843:914 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 5 28 gly glycopeptide 891:902 arg2 glycopeptide 891:902 theoretical glycopeptide 879:902 theoretical glycopeptide 879:902 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 5 29 theme precursor 960:968 arg1 mass 970:973 the query precursor mass 950:973 the query precursor mass 950:973 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 4 30 theme advanced 676:683 arg1 mode 691:694 an advanced query mode 673:694 an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters 673:795 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 2 31 theme precursor 460:468 arg1 ion 470:472 the query precursor ion 450:472 the query precursor ion 450:472 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 6 32 theme composition 1032:1042 arg1 filtering 1044:1052 glycan composition filtering 1025:1052 glycan composition filtering 1025:1052 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 7 33 theme mass 1327:1330 arg1 matches 1332:1338 the most probable intact glycopeptide mass matches 1289:1338 the most probable intact glycopeptide mass matches 1289:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 7 34 theme verification 1358:1369 arg1 process 1371:1377 the verification process 1354:1377 the verification process 1354:1377 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 4 35 theme experimental 773:784 arg1 parameters 786:795 several experimental parameters 765:795 several experimental parameters 765:795 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 6 36 theme in 1093:1094 arg1 diagram 1125:1131 an in silico peptide fragmentation diagram 1090:1131 an in silico peptide fragmentation diagram 1090:1131 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 1 37 theme manual 177:182 arg1 annotation 184:193 the manual annotation 173:193 the manual annotation of intact glycopeptides from MS data 173:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 2 38 theme query 454:458 arg1 ion 470:472 the query precursor ion 450:472 the query precursor ion 450:472 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 1 39 gly glycopeptides 205:217 arg2 glycopeptides 205:217 intact glycopeptides 198:217 intact glycopeptides from MS data 198:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 39 gly glycopeptides 205:217 arg1 data 227:230 MS data 224:230 MS data 224:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 2 40 theme exploratory 312:322 arg1 tool 324:327 This exploratory tool 307:327 EXPERIMENTAL DESIGN This exploratory tool 287:327 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 2 41 theme peptide-glycan 386:399 arg1 combinations 401:412 all peptide-glycan combinations 382:412 all peptide-glycan combinations that fall within the error range of the query precursor ion 382:472 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 0 42 theme Web-Based 16:24 arg1 Tool 26:29 A Web-Based Tool 14:29 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 0:93 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 4 43 theme several 765:771 arg1 parameters 786:795 several experimental parameters 765:795 several experimental parameters 765:795 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 3 44 theme precursor 594:602 arg1 mass 604:607 its precursor mass 590:607 its precursor mass 590:607 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 6 45 theme correct 1168:1174 arg1 identification 1189:1202 the correct glycopeptide identification 1164:1202 the correct glycopeptide identification 1164:1202 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 4 46 theme search 639:644 arg1 space 646:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 4 46 theme search 639:644 arg1 customized 659:668 customized 659:668 customized 659:668 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 6 47 dep chart 1006:1010 arg1 incorporates 1012:1023 incorporates 1012:1023 incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram 1012:1131 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 1 48 theme PURPOSE 95:101 arg1 tool 145:148 a web-based visualization tool 119:148 a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data 119:284 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 48 theme PURPOSE 95:101 arg1 PepSweetener 103:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 4 49 from customized 659:668 arg1 mode 691:694 an advanced query mode 673:694 an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters 673:795 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 1 50 theme intact 198:203 arg1 glycopeptides 205:217 intact glycopeptides 198:217 intact glycopeptides from MS data 198:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 2 51 dep DESIGN 300:305 arg1 tool 324:327 This exploratory tool 307:327 EXPERIMENTAL DESIGN This exploratory tool 287:327 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 6 52 theme peptide 1103:1109 arg1 diagram 1125:1131 an in silico peptide fragmentation diagram 1090:1131 an in silico peptide fragmentation diagram 1090:1131 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 1 53 theme glycopeptides 205:217 arg1 annotation 184:193 the manual annotation 173:193 the manual annotation of intact glycopeptides from MS data 173:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 4 54 theme potential 711:719 arg1 proteins/peptides 721:737 potential proteins/peptides 711:737 potential proteins/peptides 711:737 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 6 55 theme glycan 1025:1030 arg1 filtering 1044:1052 glycan composition filtering 1025:1052 glycan composition filtering 1025:1052 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 6 56 dep in 1093:1094 arg1 silico 1096:1101 silico 1096:1101 silico 1096:1101 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 0 57 theme Manual 42:47 arg1 Annotation 49:58 Support Manual Annotation 34:58 Support Manual Annotation of Intact Glycopeptide MS Spectra 34:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 7 58 theme probable 1298:1305 arg1 matches 1332:1338 the most probable intact glycopeptide mass matches 1289:1338 the most probable intact glycopeptide mass matches 1289:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 5 59 theme interactive 846:856 arg1 chart 867:871 an interactive heat-map chart 843:871 an interactive heat-map chart where theoretical glycopeptide tile colors 843:914 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 6 60 theme glycopeptide 1176:1187 arg1 identification 1189:1202 the correct glycopeptide identification 1164:1202 the correct glycopeptide identification 1164:1202 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 2 61 theme glycopeptide 348:359 arg1 dataset 361:367 a theoretical glycopeptide dataset 334:367 a theoretical glycopeptide dataset 334:367 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 0 62 theme Support 34:40 arg1 Annotation 49:58 Support Manual Annotation 34:58 Support Manual Annotation of Intact Glycopeptide MS Spectra 34:92 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 2 63 gly glycopeptide 348:359 arg2 glycopeptide 348:359 a theoretical glycopeptide dataset 334:367 a theoretical glycopeptide dataset 334:367 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 9 64 dep http 1540:1543 arg1 //glycoproteome.expasy.org/pepsweetener/app/ 1545:1588 //glycoproteome.expasy.org/pepsweetener/app/ 1545:1588 http://glycoproteome.expasy.org/pepsweetener/app/ 1540:1588 The tool is publicly hosted on ExPASy, the SIB Swiss Institute of Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/). 28975713 7 65 theme intact 1307:1312 arg1 matches 1332:1338 the most probable intact glycopeptide mass matches 1289:1338 the most probable intact glycopeptide mass matches 1289:1338 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 2 66 theme theoretical 336:346 arg1 dataset 361:367 a theoretical glycopeptide dataset 334:367 a theoretical glycopeptide dataset 334:367 EXPERIMENTAL DESIGN This exploratory tool uses a theoretical glycopeptide dataset to visualize all peptide-glycan combinations that fall within the error range of the query precursor ion. 28975713 6 67 theme visualization 992:1004 arg1 chart 1006:1010 a visualization chart 990:1010 a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram 990:1131 Additionally, a visualization chart incorporates glycan composition filtering, sorting by mass and tolerance, and an in silico peptide fragmentation diagram is provided to further support the correct glycopeptide identification. 28975713 9 68 dep portal 1532:1537 arg1 http 1540:1543 http 1540:1543 http://glycoproteome.expasy.org/pepsweetener/app/ 1540:1588 The tool is publicly hosted on ExPASy, the SIB Swiss Institute of Bioinformatics resource portal (http://glycoproteome.expasy.org/pepsweetener/app/). 28975713 7 69 theme RELEVANCE 1230:1238 arg1 PepSweetener 1240:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener 1205:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener 1205:1251 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 8 70 theme protein 1405:1411 arg1 samples 1413:1419 serum protein samples 1399:1419 serum protein samples 1399:1419 It is validated on serum protein samples and immunoglobulins. 28975713 1 71 theme MS 224:225 arg1 data 227:230 MS data 224:230 MS data 224:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 72 from data 227:230 arg1 annotation 184:193 the manual annotation 173:193 the manual annotation of intact glycopeptides from MS data 173:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 72 from data 227:230 arg1 glycopeptides 205:217 intact glycopeptides 198:217 intact glycopeptides from MS data 198:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 5 73 theme theoretical 879:889 arg1 glycopeptide 891:902 theoretical glycopeptide 879:902 theoretical glycopeptide 879:902 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 1 74 theme web-based 121:129 arg1 tool 145:148 a web-based visualization tool 119:148 a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data 119:284 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 74 theme web-based 121:129 arg1 PepSweetener 103:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 4 75 theme glycopeptide 626:637 arg1 space 646:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space 610:650 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 4 75 theme glycopeptide 626:637 arg1 customized 659:668 customized 659:668 customized 659:668 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 28975713 5 76 theme RESULTS 798:804 arg1 PepSweetener 806:817 RESULTS PepSweetener 798:817 RESULTS PepSweetener 798:817 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 5 77 theme query 954:958 arg1 mass 970:973 the query precursor mass 950:973 the query precursor mass 950:973 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 1 78 from annotation 184:193 arg1 data 227:230 MS data 224:230 MS data 224:230 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 79 theme visualization 131:143 arg1 tool 145:148 a web-based visualization tool 119:148 a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data 119:284 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 1 79 theme visualization 131:143 arg1 PepSweetener 103:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener 95:114 PURPOSE PepSweetener is a web-based visualization tool designed to facilitate the manual annotation of intact glycopeptides from MS data regardless of the instrument that produced these data. 28975713 5 80 theme ppm 930:932 arg1 deviations 934:943 ppm deviations 930:943 ppm deviations from the query precursor mass 930:973 RESULTS PepSweetener displays the results on an interactive heat-map chart where theoretical glycopeptide tile colors correspond to ppm deviations from the query precursor mass. 28975713 7 81 theme CLINICAL 1221:1228 arg1 RELEVANCE 1230:1238 CLINICAL RELEVANCE 1221:1238 CLINICAL RELEVANCE 1221:1238 CONCLUSIONS AND CLINICAL RELEVANCE PepSweetener efficiently allows the selection of the most probable intact glycopeptide mass matches and speeds up the verification process. 28975713 0 82 dep PepSweetener 0:11 arg1 Tool 26:29 A Web-Based Tool 14:29 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 0:93 PepSweetener: A Web-Based Tool to Support Manual Annotation of Intact Glycopeptide MS Spectra. 28975713 3 83 theme glycan 544:549 arg1 composition 551:561 the correct peptide and glycan composition 520:561 composition 551:561 PepSweetener simplifies the determination of the correct peptide and glycan composition of a glycopeptide based on its precursor mass. 28975713 4 84 theme glycan 740:745 arg1 compositions 747:758 glycan compositions 740:758 glycan compositions 740:758 The theoretical glycopeptide search space can be customized in an advanced query mode that specifies potential proteins/peptides, glycan compositions, and several experimental parameters. 25641685 6 0 theme monosialylated 1023:1036 arg1 apoC-III 1038:1045 monosialylated apoC-III 1023:1045 monosialylated apoC-III 1023:1045 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 7 1 theme glycoforms 1238:1247 arg1 analysis 1217:1224 the analysis 1213:1224 the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs 1213:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 7 2 theme RELEVANCE 1154:1162 arg1 MS 1174:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 3 3 theme CDG-associated 577:590 arg1 defects 601:607 CDG-associated O-glycan defects 577:607 CDG-associated O-glycan defects 577:607 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 4 4 theme ATP6V0A2-CDG 670:681 arg1 patients 683:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 3 5 theme O-glycan 592:599 arg1 defects 601:607 CDG-associated O-glycan defects 577:607 CDG-associated O-glycan defects 577:607 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 6 6 theme abnormal 1100:1107 arg1 structure 1118:1126 an abnormal O-glycan structure 1097:1126 an abnormal O-glycan structure 1097:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 5 7 with CDG-patients 880:891 arg1 emphasis 898:905 emphasis 898:905 emphasis to COG-CDG 898:916 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 25641685 3 8 dep detection 543:551 arg1 the 539:541 the 539:541 the 539:541 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 4 9 from patients 683:690 arg1 plasmas 630:636 EXPERIMENTAL DESIGN plasmas 610:636 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 610:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 6 10 theme particular 1000:1009 arg1 behavior 1011:1018 a particular behavior 998:1018 a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure 998:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 7 11 theme apoC-III 1229:1236 arg1 glycoforms 1238:1247 apoC-III glycoforms 1229:1247 apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs 1229:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 5 12 theme various 834:840 arg1 abnormalities 863:875 various significant O-glycan abnormalities 834:875 various significant O-glycan abnormalities 834:875 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 25641685 2 13 theme circulating 413:423 arg1 C-III 440:444 circulating apolipoprotein C-III 413:444 circulating apolipoprotein C-III 413:444 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 3 14 theme MALDI-TOF 510:518 arg1 MS 520:521 MALDI-TOF MS 510:521 MALDI-TOF MS of apoC-III 510:533 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 5 15 theme significant 842:852 arg1 abnormalities 863:875 various significant O-glycan abnormalities 834:875 various significant O-glycan abnormalities 834:875 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 25641685 2 16 theme O-glycan 170:177 arg1 abnormalities 179:191 The O-glycan abnormalities 166:191 The O-glycan abnormalities 166:191 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 6 17 theme O-glycan 1109:1116 arg1 structure 1118:1126 an abnormal O-glycan structure 1097:1126 an abnormal O-glycan structure 1097:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 3 18 theme study 469:473 arg1 objective 451:459 The objective 447:459 The objective of this study 447:473 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 3 19 theme MS 520:521 arg1 reliability 495:505 the reliability 491:505 the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects 491:607 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 1 20 with Comparison 109:118 arg1 electrophoresis 141:155 two-dimensional electrophoresis 125:155 two-dimensional electrophoresis 125:155 Comparison with two-dimensional electrophoresis. 25641685 5 21 theme O-glycan 854:861 arg1 abnormalities 863:875 various significant O-glycan abnormalities 834:875 various significant O-glycan abnormalities 834:875 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 25641685 2 22 theme electrophoresis 378:392 arg1 methods 394:400 electrophoresis methods 378:400 electrophoresis methods applied to circulating apolipoprotein C-III 378:444 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 7 23 theme ATP6V0A2-CDGs 1293:1305 arg1 screening 1271:1279 potential routine screening 1253:1279 potential routine screening of COG- and ATP6V0A2-CDGs 1253:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 0 24 theme MALDI-TOF 0:8 arg1 MS 10:11 MALDI-TOF MS 0:11 MALDI-TOF MS 0:11 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 6 25 theme CDG 935:937 arg1 samples 939:945 CDG samples 935:945 CDG samples 935:945 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 6 26 theme comparison 948:957 arg1 study 959:963 comparison study 948:963 comparison study between 2DE and MALDI-TOF 948:989 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 7 27 theme COG- 1284:1287 arg1 screening 1271:1279 potential routine screening 1253:1279 potential routine screening of COG- and ATP6V0A2-CDGs 1253:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 4 28 theme CDG-negative 643:654 arg1 patients 683:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 7 29 theme powerful 1190:1197 arg1 technique 1199:1207 a powerful technique 1188:1207 a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs 1188:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 2 30 theme apolipoprotein 425:438 arg1 C-III 440:444 circulating apolipoprotein C-III 413:444 circulating apolipoprotein C-III 413:444 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 7 31 gly glycoforms 1238:1247 arg1 apoC-III 1229:1236 apoC-III glycoforms 1229:1247 apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs 1229:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 4 32 theme COG-CDG 657:663 arg1 patients 683:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 643:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 5 33 theme apoC-III 807:814 arg1 MALDI-TOF 794:802 MALDI-TOF 794:802 RESULTS MALDI-TOF of apoC-III 786:814 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 25641685 7 34 theme potential 1253:1261 arg1 screening 1271:1279 potential routine screening 1253:1279 potential routine screening of COG- and ATP6V0A2-CDGs 1253:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 0 35 theme apoC-III 24:31 arg1 glycoforms 33:42 apoC-III glycoforms 24:42 apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation 24:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 4 36 theme DESIGN 623:628 arg1 plasmas 630:636 EXPERIMENTAL DESIGN plasmas 610:636 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 610:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 2 37 theme congenital 211:220 arg1 disorders 222:230 some congenital disorders 206:230 some congenital disorders of glycosylation 206:247 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 7 38 theme CONCLUSIONS 1129:1139 arg1 MS 1174:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 3 39 theme apoC-III 526:533 arg1 MS 520:521 MALDI-TOF MS 510:521 MALDI-TOF MS of apoC-III 510:533 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 0 40 theme patients 47:54 arg1 glycoforms 33:42 apoC-III glycoforms 24:42 apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation 24:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 4 41 theme EXPERIMENTAL 610:621 arg1 plasmas 630:636 EXPERIMENTAL DESIGN plasmas 610:636 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients 610:690 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 7 42 theme MALDI-TOF 1164:1172 arg1 MS 1174:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS 1129:1175 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 2 43 theme Golgi-congenital 278:293 arg1 disorders 295:303 namely conserved oligomeric Golgi-congenital disorders 250:303 namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) 250:331 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 1 44 theme two-dimensional 125:139 arg1 electrophoresis 141:155 two-dimensional electrophoresis 125:155 two-dimensional electrophoresis 125:155 Comparison with two-dimensional electrophoresis. 25641685 7 45 theme routine 1263:1269 arg1 screening 1271:1279 potential routine screening 1253:1279 potential routine screening of COG- and ATP6V0A2-CDGs 1253:1305 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 2 46 dep PURPOSE 158:164 arg1 detected 363:370 detected 363:370 are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III 352:444 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 47 theme conserved 257:265 arg1 disorders 295:303 namely conserved oligomeric Golgi-congenital disorders 250:303 namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) 250:331 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 48 theme oligomeric 267:276 arg1 disorders 295:303 namely conserved oligomeric Golgi-congenital disorders 250:303 namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) 250:331 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 7 49 theme CLINICAL 1145:1152 arg1 RELEVANCE 1154:1162 CLINICAL RELEVANCE 1145:1162 CLINICAL RELEVANCE 1145:1162 CONCLUSIONS AND CLINICAL RELEVANCE MALDI-TOF MS appears as a powerful technique for the analysis of apoC-III glycoforms for potential routine screening of COG- and ATP6V0A2-CDGs. 25641685 6 50 theme mass 1054:1057 arg1 spectrometer 1059:1070 the mass spectrometer 1050:1070 the mass spectrometer that could be related to an abnormal O-glycan structure 1050:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 6 50 theme mass 1054:1057 arg1 related 1086:1092 related 1086:1092 related 1086:1092 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 0 51 gly glycoforms 33:42 arg1 apoC-III 24:31 apoC-III glycoforms 24:42 apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation 24:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 0 51 gly glycoforms 33:42 arg1 patients 47:54 patients 47:54 patients with congenital disorders affecting O-glycosylation 47:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 0 52 with patients 47:54 arg1 disorders 72:80 congenital disorders 61:80 congenital disorders affecting O-glycosylation 61:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 0 53 theme congenital 61:70 arg1 disorders 72:80 congenital disorders 61:80 congenital disorders affecting O-glycosylation 61:106 MALDI-TOF MS applied to apoC-III glycoforms of patients with congenital disorders affecting O-glycosylation. 25641685 2 54 theme glycosylation 235:247 arg1 disorders 222:230 some congenital disorders 206:230 some congenital disorders of glycosylation 206:247 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 54 theme glycosylation 235:247 arg1 disorders 295:303 namely conserved oligomeric Golgi-congenital disorders 250:303 namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) 250:331 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 54 theme glycosylation 235:247 arg1 ATP6V0A2-CDGs 337:349 ATP6V0A2-CDGs 337:349 ATP6V0A2-CDGs 337:349 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 3 55 theme defects 601:607 arg1 characterization 557:572 characterization 557:572 characterization 557:572 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 3 55 theme defects 601:607 arg1 detection 543:551 detection 543:551 detection 543:551 The objective of this study was to evaluate the reliability of MALDI-TOF MS of apoC-III for the detection and characterization of CDG-associated O-glycan defects. 25641685 6 56 from behavior 1011:1018 arg1 spectrometer 1059:1070 the mass spectrometer 1050:1070 the mass spectrometer that could be related to an abnormal O-glycan structure 1050:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 6 56 from behavior 1011:1018 arg1 related 1086:1092 related 1086:1092 related 1086:1092 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 6 57 theme apoC-III 1038:1045 arg1 behavior 1011:1018 a particular behavior 998:1018 a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure 998:1126 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 2 58 theme glycosylation 308:320 arg1 disorders 222:230 some congenital disorders 206:230 some congenital disorders of glycosylation 206:247 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 58 theme glycosylation 308:320 arg1 disorders 295:303 namely conserved oligomeric Golgi-congenital disorders 250:303 namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) 250:331 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 2 58 theme glycosylation 308:320 arg1 ATP6V0A2-CDGs 337:349 ATP6V0A2-CDGs 337:349 ATP6V0A2-CDGs 337:349 PURPOSE The O-glycan abnormalities accompanying some congenital disorders of glycosylation, namely conserved oligomeric Golgi-congenital disorders of glycosylation (COG-CDGs) and ATP6V0A2-CDGs, are mainly detected using electrophoresis methods applied to circulating apolipoprotein C-III. 25641685 4 59 theme Western 772:778 arg1 blot 780:783 Western blot 772:783 Western blot 772:783 EXPERIMENTAL DESIGN plasmas from CDG-negative, COG-CDG, and ATP6V0A2-CDG patients were analyzed and results were compared to those obtained using 2DE followed by Western blot. 25641685 6 60 gly monosialylated 1023:1036 arg1 apoC-III 1038:1045 monosialylated apoC-III 1023:1045 monosialylated apoC-III 1023:1045 Furthermore, in CDG samples, comparison study between 2DE and MALDI-TOF showed a particular behavior of monosialylated apoC-III in the mass spectrometer that could be related to an abnormal O-glycan structure. 25641685 5 61 dep RESULTS 786:792 arg1 MALDI-TOF 794:802 MALDI-TOF 794:802 RESULTS MALDI-TOF of apoC-III 786:814 RESULTS MALDI-TOF of apoC-III allowed to detect various significant O-glycan abnormalities in CDG-patients with emphasis to COG-CDG. 27873468 0 0 theme Structure/Density/Position 83:108 arg1 Effects 65:71 Effects 65:71 Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays 65:179 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 6 1 theme library 1283:1289 arg1 variants 1365:1372 rationally engineered variants 1343:1372 rationally engineered variants of the lectins 1343:1387 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 1 theme library 1283:1289 arg1 elaboration 1251:1261 further synthetic elaboration 1233:1261 further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins 1233:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 1 2 with pairing 193:199 arg1 lectins 241:247 tissue lectins 234:247 tissue lectins 234:247 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 4 3 theme negative 1056:1063 arg1 which 1032:1036 which 1032:1036 which 1032:1036 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 4 3 theme negative 1056:1063 arg1 control 1065:1071 a negative control 1054:1071 a negative control 1054:1071 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 1 4 theme determinative 286:298 arg1 factors 300:306 determinative factors 286:306 determinative factors 286:306 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 0 5 theme Glycan 76:81 arg1 Structure/Density/Position 83:108 Glycan Structure/Density/Position 76:108 Glycan Structure/Density/Position 76:108 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 5 6 theme core 1154:1157 arg1 sialylation 1161:1171 core 1 sialylation 1154:1171 core 1 sialylation 1154:1171 Remarkably, the two tandem-repeat-type galectins-4 and -8 were distinguished by core 1 sialylation, as the two separated domains were. 27873468 5 7 theme tandem-repeat-type 1094:1111 arg1 galectins-4 1113:1123 the two tandem-repeat-type galectins-4 1086:1123 the two tandem-repeat-type galectins-4 1086:1123 Remarkably, the two tandem-repeat-type galectins-4 and -8 were distinguished by core 1 sialylation, as the two separated domains were. 27873468 6 8 theme network 1310:1316 arg1 library 1283:1289 glycopeptide library 1270:1289 glycopeptide library 1270:1289 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 8 theme network 1310:1316 arg1 testing 1295:1301 testing 1295:1301 testing 1295:1301 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 3 9 theme array-based 726:736 arg1 screening 738:746 an array-based screening 723:746 an array-based screening of tissue lectins 723:764 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 6 10 theme testing 1295:1301 arg1 variants 1365:1372 rationally engineered variants 1343:1372 rationally engineered variants of the lectins 1343:1387 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 10 theme testing 1295:1301 arg1 elaboration 1251:1261 further synthetic elaboration 1233:1261 further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins 1233:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 3 11 theme peptide 555:561 arg1 motif 579:583 a peptide repeat sequence motif 553:583 a peptide repeat sequence motif of the glycoprotein mucin-1 553:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 4 12 theme galectin-related 1006:1021 arg1 protein 1023:1029 the galectin-related protein 1002:1029 the galectin-related protein 1002:1029 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 6 13 theme lectins 1381:1387 arg1 variants 1365:1372 rationally engineered variants 1343:1372 rationally engineered variants of the lectins 1343:1387 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 13 theme lectins 1381:1387 arg1 elaboration 1251:1261 further synthetic elaboration 1233:1261 further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins 1233:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 14 dep library 1283:1289 arg1 the 1266:1268 the 1266:1268 the 1266:1268 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 1 15 theme cellular 204:211 arg1 glycoconjugates 213:227 cellular glycoconjugates 204:227 cellular glycoconjugates 204:227 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 2 16 theme responsiveness 514:527 arg1 degrees 503:509 degrees 503:509 degrees of responsiveness to these factors 503:544 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 4 17 from signals 878:884 arg1 range 859:863 the range 855:863 the range from intense signals 855:884 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 4 18 theme intense 870:876 arg1 signals 878:884 intense signals 870:884 intense signals 870:884 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 2 19 theme different 453:461 arg1 members 463:469 different members 453:469 different members of a lectin family 453:488 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 1 20 theme glycoconjugates 213:227 arg1 pairing 193:199 Functional pairing 182:199 Functional pairing of cellular glycoconjugates with tissue lectins 182:247 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 1 20 theme glycoconjugates 213:227 arg1 process 271:277 a highly selective process 252:277 a highly selective process 252:277 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 0 21 theme Mucin-Like 10:19 arg1 Glycopeptides 21:33 Synthetic Mucin-Like Glycopeptides 0:33 Synthetic Mucin-Like Glycopeptides as Versatile Tools 0:52 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 3 22 from variations 683:692 arg1 parameters 709:718 these three parameters 697:718 these three parameters 697:718 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 6 23 theme glycopeptide 1270:1281 arg1 library 1283:1289 glycopeptide library 1270:1289 glycopeptide library 1270:1289 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 0 24 with Interaction 117:127 arg1 Galectins 161:169 Adhesion/Growth-Regulatory Galectins 134:169 Adhesion/Growth-Regulatory Galectins in Arrays 134:179 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 0 25 theme Synthetic 0:8 arg1 Glycopeptides 21:33 Synthetic Mucin-Like Glycopeptides 0:33 Synthetic Mucin-Like Glycopeptides as Versatile Tools 0:52 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 3 26 theme lectins 758:764 arg1 screening 738:746 an array-based screening 723:746 an array-based screening of tissue lectins 723:764 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 1 27 theme tissue 234:239 arg1 lectins 241:247 tissue lectins 234:247 tissue lectins 234:247 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 5 28 theme separated 1185:1193 arg1 domains 1195:1201 the two separated domains 1177:1201 the two separated domains 1177:1201 Remarkably, the two tandem-repeat-type galectins-4 and -8 were distinguished by core 1 sialylation, as the two separated domains were. 27873468 6 29 theme further 1233:1239 arg1 elaboration 1251:1261 further synthetic elaboration 1233:1261 further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins 1233:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 2 30 theme Glycan 344:349 arg1 structure 351:359 Glycan structure 344:359 Glycan structure 344:359 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 0 31 theme Adhesion/Growth-Regulatory 134:159 arg1 Galectins 161:169 Adhesion/Growth-Regulatory Galectins 134:169 Adhesion/Growth-Regulatory Galectins in Arrays 134:179 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 3 32 theme glyco 654:658 arg1 peptides 660:667 synthetic (glyco)peptides 643:667 synthetic (glyco)peptides 643:667 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 3 33 theme sequence 570:577 arg1 motif 579:583 a peptide repeat sequence motif 553:583 a peptide repeat sequence motif of the glycoprotein mucin-1 553:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 2 34 theme family 483:488 arg1 members 463:469 different members 453:469 different members of a lectin family 453:488 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 3 35 theme tissue 751:756 arg1 lectins 758:764 tissue lectins 751:764 tissue lectins 751:764 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 6 36 theme engineered 1354:1363 arg1 variants 1365:1372 rationally engineered variants 1343:1372 rationally engineered variants of the lectins 1343:1387 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 3 37 theme mucin-1 605:611 arg1 motif 579:583 a peptide repeat sequence motif 553:583 a peptide repeat sequence motif of the glycoprotein mucin-1 553:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 3 38 gly glycoprotein 592:603 arg1 glycoprotein 592:603 the glycoprotein mucin-1 588:611 the glycoprotein mucin-1 588:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 2 39 theme lectin 476:481 arg1 family 483:488 a lectin family 474:488 a lectin family 474:488 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 0 40 theme Versatile 38:46 arg1 Tools 48:52 Versatile Tools 38:52 Versatile Tools 38:52 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 3 41 theme synthetic 643:651 arg1 peptides 660:667 synthetic (glyco)peptides 643:667 synthetic (glyco)peptides 643:667 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 4 42 theme binding 970:976 arg1 lack 962:965 a lack 960:965 a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control 960:1071 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 0 43 from Effects 65:71 arg1 Interaction 117:127 the Interaction 113:127 the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays 113:179 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 6 44 theme synthetic 1241:1249 arg1 elaboration 1251:1261 further synthetic elaboration 1233:1261 further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins 1233:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 6 45 theme galectins 1329:1337 arg1 network 1310:1316 the network 1306:1316 the network of natural galectins 1306:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 3 46 theme distinct 674:681 arg1 variations 683:692 distinct variations 674:692 distinct variations in these three parameters 674:718 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 3 47 theme glycoprotein 592:603 arg1 mucin-1 605:611 the glycoprotein mucin-1 588:611 the glycoprotein mucin-1 588:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 2 48 theme presentation 374:385 arg1 modes 365:369 modes 365:369 modes 365:369 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 2 48 theme presentation 374:385 arg1 structure 351:359 Glycan structure 344:359 Glycan structure 344:359 Glycan structure and modes of presentation, that is, its position and density, can contribute to binding, as different members of a lectin family can regulate degrees of responsiveness to these factors. 27873468 6 49 theme natural 1321:1327 arg1 galectins 1329:1337 natural galectins 1321:1337 natural galectins 1321:1337 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 3 50 theme repeat 563:568 arg1 motif 579:583 a peptide repeat sequence motif 553:583 a peptide repeat sequence motif of the glycoprotein mucin-1 553:611 Using a peptide repeat sequence motif of the glycoprotein mucin-1, the principle of introducing synthetic (glyco)peptides with distinct variations in these three parameters to an array-based screening of tissue lectins is illustrated. 27873468 1 51 theme selective 261:269 arg1 pairing 193:199 Functional pairing 182:199 Functional pairing of cellular glycoconjugates with tissue lectins 182:247 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 1 51 theme selective 261:269 arg1 process 271:277 a highly selective process 252:277 a highly selective process 252:277 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 4 52 theme Interaction 782:792 arg1 profiles 794:801 Interaction profiles 782:801 Interaction profiles of seven adhesion/growth-regulatory galectins 782:847 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 6 53 gly glycopeptide 1270:1281 arg2 glycopeptide 1270:1281 glycopeptide library 1270:1289 glycopeptide library 1270:1289 These results encourage further synthetic elaboration of the glycopeptide library and testing of the network of natural galectins and rationally engineered variants of the lectins. 27873468 4 54 theme galectins 839:847 arg1 profiles 794:801 Interaction profiles 782:801 Interaction profiles of seven adhesion/growth-regulatory galectins 782:847 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 4 55 dep core 891:894 arg1 pentasaccharides 898:913 2 pentasaccharides 896:913 2 pentasaccharides 896:913 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 0 56 from Galectins 161:169 arg1 Arrays 174:179 Arrays 174:179 Arrays 174:179 Synthetic Mucin-Like Glycopeptides as Versatile Tools to Measure Effects of Glycan Structure/Density/Position on the Interaction with Adhesion/Growth-Regulatory Galectins in Arrays. 27873468 4 57 theme adhesion/growth-regulatory 812:837 arg1 galectins 839:847 seven adhesion/growth-regulatory galectins 806:847 seven adhesion/growth-regulatory galectins 806:847 Interaction profiles of seven adhesion/growth-regulatory galectins cover the range from intense signals with core 2 pentasaccharides and core 1 binding for galectins-3 and -5 to a lack of binding for galectin-1 and also the galectin-related protein, which was included as a negative control. 27873468 1 58 theme Functional 182:191 arg1 pairing 193:199 Functional pairing 182:199 Functional pairing of cellular glycoconjugates with tissue lectins 182:247 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 27873468 1 58 theme Functional 182:191 arg1 process 271:277 a highly selective process 252:277 a highly selective process 252:277 Functional pairing of cellular glycoconjugates with tissue lectins is a highly selective process, whose determinative factors have not yet been fully delineated. 24085030 7 0 theme glycan 1203:1208 arg1 structures 1210:1219 altered glycan structures 1195:1219 altered glycan structures 1195:1219 The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. 24085030 4 1 with ELISA 700:704 arg1 lectins 735:741 the lectins 731:741 the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans 731:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 3 2 theme salivary 473:480 arg1 glycoproteins 482:494 salivary glycoproteins 473:494 salivary glycoproteins of alcoholics 473:508 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 9 3 theme salivary 1396:1403 arg1 glycoproteins 1405:1417 the salivary glycoproteins 1392:1417 the salivary glycoproteins of alcohol-dependent people 1392:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 7 4 theme disease-related 1089:1103 arg1 alterations 1105:1115 disease-related alterations 1089:1115 disease-related alterations 1089:1115 The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. 24085030 4 5 theme T 817:817 arg1 antigens 826:833 T and Tn antigens 817:833 antigens 826:833 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 4 6 theme healthy 651:657 arg1 controls 659:666 21 healthy controls 648:666 21 healthy controls 648:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 10 7 theme light 1544:1548 arg1 chains 1550:1555 heavy and light chains 1534:1555 heavy and light chains of immunoglobulins 1534:1574 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 9 8 theme people 1440:1445 arg1 glycoproteins 1405:1417 the salivary glycoproteins 1392:1417 the salivary glycoproteins of alcohol-dependent people 1392:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 4 9 theme Tn 823:824 arg1 antigens 826:833 T and Tn antigens 817:833 antigens 826:833 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 1 10 theme carbohydrate 181:192 arg1 transferrin 204:214 carbohydrate deficient transferrin 181:214 carbohydrate deficient transferrin (CDT) 181:220 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 1 10 theme carbohydrate 181:192 arg1 CDT 217:219 CDT 217:219 CDT 217:219 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 4 11 theme alcohol-dependent 617:633 arg1 patients 635:642 31 alcohol-dependent patients 614:642 31 alcohol-dependent patients 614:642 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 3 12 theme efficient 531:539 arg1 carriers 541:548 the most efficient carriers 522:548 the most efficient carriers of such marker glycoepitopes 522:577 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 5 13 gly fucosylation 886:897 arg1 T-antigen 935:943 T-antigen 935:943 T-antigen 935:943 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 11 14 theme intake 1789:1794 arg1 intake 1823:1828 alcohol intake 1815:1828 alcohol intake 1815:1828 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 14 theme intake 1789:1794 arg1 amount 1771:1776 the amount 1767:1776 the amount of alcohol intake 1767:1794 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 14 theme intake 1789:1794 arg1 intake 1789:1794 alcohol intake 1781:1794 alcohol intake 1781:1794 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 14 theme intake 1789:1794 arg1 duration 1803:1810 the duration 1799:1810 the duration of alcohol intake 1799:1828 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 3 15 theme marker 558:563 arg1 glycoepitopes 565:577 such marker glycoepitopes 553:577 such marker glycoepitopes 553:577 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 4 16 from acid 801:804 arg1 O-glycans 838:846 O-glycans 838:846 O-glycans 838:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 1 17 theme serum 119:123 arg1 proteins 125:132 serum proteins 119:132 serum proteins 119:132 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 8 18 theme glycosylation 1272:1284 arg1 profile 1286:1292 the glycosylation profile 1268:1292 the glycosylation profile 1268:1292 In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. 24085030 1 19 theme proteins 125:132 arg1 Glycosylation 102:114 AIM Glycosylation 98:114 AIM Glycosylation of serum proteins 98:132 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 7 20 theme altered 1195:1201 arg1 structures 1210:1219 altered glycan structures 1195:1219 altered glycan structures 1195:1219 The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. 24085030 4 21 from fucose 775:780 arg1 O-glycans 838:846 O-glycans 838:846 O-glycans 838:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 1 22 theme heavy 161:165 arg1 drinking 167:174 prolonged heavy drinking 151:174 prolonged heavy drinking 151:174 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 4 23 gly glycoproteins 597:609 arg1 glycoproteins 597:609 Salivary glycoproteins 588:609 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls 580:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 4 24 theme lectin 710:715 arg1 blotting 717:724 lectin blotting 710:724 lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans 710:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 10 25 theme salivary 1467:1474 arg1 transferrin 1581:1591 transferrin 1581:1591 transferrin 1581:1591 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 25 theme salivary 1467:1474 arg1 clusterin 1510:1518 clusterin 1510:1518 clusterin 1510:1518 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 25 theme salivary 1467:1474 arg1 chains 1550:1555 heavy and light chains 1534:1555 heavy and light chains of immunoglobulins 1534:1574 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 25 theme salivary 1467:1474 arg1 glycoproteins 1476:1488 salivary glycoproteins 1467:1488 salivary glycoproteins 1467:1488 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 25 theme salivary 1467:1474 arg1 haptoglobin 1521:1531 haptoglobin 1521:1531 haptoglobin 1521:1531 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 25 theme salivary 1467:1474 arg1 α-amylase 1499:1507 α-amylase 1499:1507 α-amylase 1499:1507 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 8 26 theme abstinence 1324:1333 arg1 weeks 1315:1319 7 weeks 1313:1319 7 weeks of abstinence 1313:1333 In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. 24085030 10 27 theme analysis 1638:1645 arg1 worthy 1605:1610 worthy 1605:1610 worthy of detailed glycosylation analysis 1605:1645 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 0 28 dep study 91:95 arg1 changes 14:20 Glycosylation changes 0:20 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients 0:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 10 29 theme detailed 1615:1622 arg1 analysis 1638:1645 detailed glycosylation analysis 1615:1645 detailed glycosylation analysis 1615:1645 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 0 30 theme Glycosylation 0:12 arg1 changes 14:20 Glycosylation changes 0:20 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients 0:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 5 31 theme α2,3 900:903 arg1 sialylation 905:915 α2,3 sialylation 900:915 α2,3 sialylation 900:915 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 1 32 theme alcohol 285:291 arg1 consumption 293:303 sustained alcohol consumption 275:303 sustained alcohol consumption 275:303 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 6 33 theme lectin 1020:1025 arg1 blotting 1027:1034 lectin blotting 1020:1034 lectin blotting 1020:1034 In lectin blotting ten glycoprotein bands were analyzed. 24085030 11 34 theme Further 1687:1693 arg1 studies 1695:1701 Further studies 1687:1701 Further studies 1687:1701 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 0 35 theme salivary 29:36 arg1 glycoproteins 38:50 the salivary glycoproteins 25:50 the salivary glycoproteins of alcohol-dependent patients 25:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 0 36 theme patients 73:80 arg1 glycoproteins 38:50 the salivary glycoproteins 25:50 the salivary glycoproteins of alcohol-dependent patients 25:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 2 37 theme acid 338:341 arg1 amount 321:326 total amount 315:326 total amount of sialic acid 315:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 37 theme acid 338:341 arg1 acid 338:341 sialic acid 331:341 sialic acid 331:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 37 theme acid 338:341 arg1 glycoepitope 359:370 the only glycoepitope 350:370 the only glycoepitope that may be altered as a result of the disease 350:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 5 38 theme patients 1007:1014 arg1 saliva 979:984 the saliva 975:984 the saliva of alcohol-dependent patients 975:1014 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 10 39 theme dependence 1675:1684 arg1 detection 1654:1662 the detection 1650:1662 the detection of alcohol dependence 1650:1684 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 0 40 gly glycoproteins 38:50 arg1 glycoproteins 38:50 the salivary glycoproteins 25:50 the salivary glycoproteins of alcohol-dependent patients 25:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 4 41 theme α2,3-bound 783:792 arg1 acid 801:804 α2,3-bound sialic acid 783:804 α2,3-bound sialic acid 783:804 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 2 42 theme only 354:357 arg1 glycoepitope 359:370 the only glycoepitope 350:370 the only glycoepitope that may be altered as a result of the disease 350:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 42 theme only 354:357 arg1 acid 338:341 sialic acid 331:341 sialic acid 331:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 42 theme only 354:357 arg1 amount 321:326 total amount 315:326 total amount of sialic acid 315:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 42 theme only 354:357 arg1 result 397:402 a result 395:402 a result of the disease 395:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 7 43 theme alterations 1105:1115 arg1 complex 1133:1139 complex 1133:1139 complex 1133:1139 The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. 24085030 7 43 theme alterations 1105:1115 arg1 profile 1078:1084 The profile 1074:1084 The profile of disease-related alterations 1074:1115 The profile of disease-related alterations was found to be complex, but all six lectins studied here were able to detect altered glycan structures. 24085030 10 44 theme immunoglobulins 1560:1574 arg1 transferrin 1581:1591 transferrin 1581:1591 transferrin 1581:1591 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 44 theme immunoglobulins 1560:1574 arg1 chains 1550:1555 heavy and light chains 1534:1555 heavy and light chains of immunoglobulins 1534:1574 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 44 theme immunoglobulins 1560:1574 arg1 haptoglobin 1521:1531 haptoglobin 1521:1531 haptoglobin 1521:1531 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 44 theme immunoglobulins 1560:1574 arg1 clusterin 1510:1518 clusterin 1510:1518 clusterin 1510:1518 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 44 theme immunoglobulins 1560:1574 arg1 α-amylase 1499:1507 α-amylase 1499:1507 α-amylase 1499:1507 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 1 45 theme specific 253:260 arg1 biomarker 262:270 well established and highly specific biomarker 225:270 well established and highly specific biomarker of sustained alcohol consumption 225:303 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 11 46 theme alcohol 1815:1821 arg1 intake 1823:1828 alcohol intake 1815:1828 alcohol intake 1815:1828 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 5 47 theme direct 860:865 arg1 ELISA 874:878 direct lectin ELISA 860:878 direct lectin ELISA 860:878 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 11 48 theme intake 1823:1828 arg1 intake 1823:1828 alcohol intake 1815:1828 alcohol intake 1815:1828 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 48 theme intake 1823:1828 arg1 amount 1771:1776 the amount 1767:1776 the amount of alcohol intake 1767:1794 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 48 theme intake 1823:1828 arg1 intake 1789:1794 alcohol intake 1781:1794 alcohol intake 1781:1794 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 11 48 theme intake 1823:1828 arg1 duration 1803:1810 the duration 1799:1810 the duration of alcohol intake 1799:1828 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 3 49 theme alcoholics 499:508 arg1 glycoproteins 482:494 salivary glycoproteins 473:494 salivary glycoproteins of alcoholics 473:508 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 9 50 theme alcohol-dependent 1422:1438 arg1 people 1440:1445 alcohol-dependent people 1422:1445 alcohol-dependent people 1422:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 10 51 from worthy 1605:1610 arg1 detection 1654:1662 the detection 1650:1662 the detection of alcohol dependence 1650:1684 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 2 52 theme total 315:319 arg1 amount 321:326 total amount 315:326 total amount of sialic acid 315:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 52 theme total 315:319 arg1 acid 338:341 sialic acid 331:341 sialic acid 331:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 52 theme total 315:319 arg1 glycoepitope 359:370 the only glycoepitope 350:370 the only glycoepitope that may be altered as a result of the disease 350:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 4 53 theme controls 659:666 arg1 glycoproteins 597:609 Salivary glycoproteins 588:609 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls 580:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 11 54 theme alcohol 1781:1787 arg1 intake 1789:1794 alcohol intake 1781:1794 alcohol intake 1781:1794 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 1 55 theme deficient 194:202 arg1 transferrin 204:214 carbohydrate deficient transferrin 181:214 carbohydrate deficient transferrin (CDT) 181:220 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 1 55 theme deficient 194:202 arg1 CDT 217:219 CDT 217:219 CDT 217:219 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 5 56 theme lectin 867:872 arg1 ELISA 874:878 direct lectin ELISA 860:878 direct lectin ELISA 860:878 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 10 57 gly glycoproteins 1476:1488 arg1 transferrin 1581:1591 transferrin 1581:1591 transferrin 1581:1591 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 57 gly glycoproteins 1476:1488 arg1 clusterin 1510:1518 clusterin 1510:1518 clusterin 1510:1518 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 57 gly glycoproteins 1476:1488 arg1 chains 1550:1555 heavy and light chains 1534:1555 heavy and light chains of immunoglobulins 1534:1574 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 57 gly glycoproteins 1476:1488 arg1 glycoproteins 1476:1488 salivary glycoproteins 1467:1488 salivary glycoproteins 1467:1488 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 57 gly glycoproteins 1476:1488 arg1 haptoglobin 1521:1531 haptoglobin 1521:1531 haptoglobin 1521:1531 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 10 57 gly glycoproteins 1476:1488 arg1 α-amylase 1499:1507 α-amylase 1499:1507 α-amylase 1499:1507 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 3 58 theme glycoepitopes 565:577 arg1 carriers 541:548 the most efficient carriers 522:548 the most efficient carriers of such marker glycoepitopes 522:577 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 4 59 theme patients 635:642 arg1 glycoproteins 597:609 Salivary glycoproteins 588:609 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls 580:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 3 60 theme such 553:556 arg1 glycoepitopes 565:577 such marker glycoepitopes 553:577 such marker glycoepitopes 553:577 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 4 61 theme lectin 693:698 arg1 ELISA 700:704 lectin ELISA 693:704 lectin ELISA 693:704 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 4 62 with blotting 717:724 arg1 lectins 735:741 the lectins 731:741 the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans 731:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 8 63 gly glycoproteins 1230:1242 arg1 glycoproteins 1230:1242 some glycoproteins 1225:1242 some glycoproteins 1225:1242 In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. 24085030 5 64 theme core 881:884 arg1 fucosylation 886:897 core fucosylation 881:897 core fucosylation 881:897 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 11 65 theme such 1732:1735 arg1 glycomarkers 1737:1748 such glycomarkers 1732:1748 such glycomarkers 1732:1748 Further studies may allow one to estimate if such glycomarkers may also reflect the amount of alcohol intake or the duration of alcohol intake. 24085030 4 66 dep METHODS 580:586 arg1 glycoproteins 597:609 Salivary glycoproteins 588:609 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls 580:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 0 67 theme pilot 85:89 arg1 study 91:95 a pilot study 83:95 a pilot study 83:95 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 9 68 from Alterations 1347:1357 arg1 profiles 1380:1387 the glycosylation profiles 1362:1387 the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people 1362:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 1 69 theme prolonged 151:159 arg1 drinking 167:174 prolonged heavy drinking 151:174 prolonged heavy drinking 151:174 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 0 70 from changes 14:20 arg1 glycoproteins 38:50 the salivary glycoproteins 25:50 the salivary glycoproteins of alcohol-dependent patients 25:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 4 71 from antigens 826:833 arg1 O-glycans 838:846 O-glycans 838:846 O-glycans 838:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 5 72 from RESULTS 849:855 arg1 sialylation 905:915 α2,3 sialylation 900:915 α2,3 sialylation 900:915 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 72 from RESULTS 849:855 arg1 expression 921:930 expression 921:930 expression of T-antigen 921:943 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 72 from RESULTS 849:855 arg1 ELISA 874:878 direct lectin ELISA 860:878 direct lectin ELISA 860:878 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 72 from RESULTS 849:855 arg1 fucosylation 886:897 core fucosylation 881:897 core fucosylation 881:897 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 3 73 gly glycoproteins 482:494 arg1 glycoproteins 482:494 salivary glycoproteins 473:494 salivary glycoproteins of alcoholics 473:508 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 4 74 theme specific 743:750 arg1 lectins 735:741 the lectins 731:741 the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans 731:846 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 10 75 theme alcohol 1667:1673 arg1 dependence 1675:1684 alcohol dependence 1667:1684 alcohol dependence 1667:1684 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 1 76 gly Glycosylation 102:114 arg1 proteins 125:132 serum proteins 119:132 serum proteins 119:132 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 5 77 theme T-antigen 935:943 arg1 sialylation 905:915 α2,3 sialylation 900:915 α2,3 sialylation 900:915 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 77 theme T-antigen 935:943 arg1 expression 921:930 expression 921:930 expression of T-antigen 921:943 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 77 theme T-antigen 935:943 arg1 ELISA 874:878 direct lectin ELISA 860:878 direct lectin ELISA 860:878 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 77 theme T-antigen 935:943 arg1 fucosylation 886:897 core fucosylation 881:897 core fucosylation 881:897 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 5 78 gly sialylation 905:915 arg1 T-antigen 935:943 T-antigen 935:943 T-antigen 935:943 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 4 79 theme core 756:759 arg1 fucose 775:780 core and antennary fucose 756:780 fucose 775:780 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 10 80 theme glycosylation 1624:1636 arg1 analysis 1638:1645 detailed glycosylation analysis 1615:1645 detailed glycosylation analysis 1615:1645 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 6 81 theme glycoprotein 1040:1051 arg1 bands 1053:1057 ten glycoprotein bands 1036:1057 ten glycoprotein bands 1036:1057 In lectin blotting ten glycoprotein bands were analyzed. 24085030 2 82 theme disease 411:417 arg1 glycoepitope 359:370 the only glycoepitope 350:370 the only glycoepitope that may be altered as a result of the disease 350:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 2 82 theme disease 411:417 arg1 result 397:402 a result 395:402 a result of the disease 395:417 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 1 83 theme sustained 275:283 arg1 consumption 293:303 sustained alcohol consumption 275:303 sustained alcohol consumption 275:303 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 1 84 theme AIM 98:100 arg1 Glycosylation 102:114 AIM Glycosylation 98:114 AIM Glycosylation of serum proteins 98:132 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 1 85 theme consumption 293:303 arg1 biomarker 262:270 well established and highly specific biomarker 225:270 well established and highly specific biomarker of sustained alcohol consumption 225:303 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 24085030 4 86 theme sialic 794:799 arg1 acid 801:804 α2,3-bound sialic acid 783:804 α2,3-bound sialic acid 783:804 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 0 87 theme alcohol-dependent 55:71 arg1 patients 73:80 alcohol-dependent patients 55:80 alcohol-dependent patients 55:80 Glycosylation changes in the salivary glycoproteins of alcohol-dependent patients: a pilot study. 24085030 9 88 theme CONCLUSION 1336:1345 arg1 Alterations 1347:1357 CONCLUSION Alterations 1336:1357 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people 1336:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 9 89 from profiles 1380:1387 arg1 glycoproteins 1405:1417 the salivary glycoproteins 1392:1417 the salivary glycoproteins of alcohol-dependent people 1392:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 5 90 theme alcohol-dependent 989:1005 arg1 patients 1007:1014 alcohol-dependent patients 989:1014 alcohol-dependent patients 989:1014 RESULTS In direct lectin ELISA, core fucosylation, α2,3 sialylation and expression of T-antigen were significantly lowered in the saliva of alcohol-dependent patients. 24085030 2 91 theme sialic 331:336 arg1 acid 338:341 sialic acid 331:341 sialic acid 331:341 However, total amount of sialic acid is not the only glycoepitope that may be altered as a result of the disease. 24085030 8 92 located observed 1298:1305 arg2 tendency 1248:1255 the tendency to correct the glycosylation profile 1244:1292 the tendency to correct the glycosylation profile 1244:1292 In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. 24085030 8 92 located observed 1298:1305 arg1 glycoproteins 1230:1242 some glycoproteins 1225:1242 some glycoproteins 1225:1242 In some glycoproteins the tendency to correct the glycosylation profile was observed after 7 weeks of abstinence. 24085030 4 93 theme Salivary 588:595 arg1 glycoproteins 597:609 Salivary glycoproteins 588:609 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls 580:666 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 6 94 gly glycoprotein 1040:1051 arg1 glycoprotein 1040:1051 ten glycoprotein bands 1036:1057 ten glycoprotein bands 1036:1057 In lectin blotting ten glycoprotein bands were analyzed. 24085030 4 95 theme antennary 765:773 arg1 fucose 775:780 core and antennary fucose 756:780 fucose 775:780 METHODS Salivary glycoproteins of 31 alcohol-dependent patients and 21 healthy controls were studied by means of lectin ELISA and lectin blotting with the lectins specific for core and antennary fucose, α2,3-bound sialic acid as well as T and Tn antigens in O-glycans. 24085030 3 96 theme glycan 444:449 arg1 structures 451:460 glycan structures 444:460 glycan structures altered in salivary glycoproteins of alcoholics 444:508 This work is focused on glycan structures altered in salivary glycoproteins of alcoholics, indicating the most efficient carriers of such marker glycoepitopes. 24085030 9 97 theme glycosylation 1366:1378 arg1 profiles 1380:1387 the glycosylation profiles 1362:1387 the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people 1362:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 10 98 theme heavy 1534:1538 arg1 chains 1550:1555 heavy and light chains 1534:1555 heavy and light chains of immunoglobulins 1534:1574 Some of salivary glycoproteins, such as α-amylase, clusterin, haptoglobin, heavy and light chains of immunoglobulins, and transferrin, seem to be worthy of detailed glycosylation analysis in the detection of alcohol dependence. 24085030 9 99 gly glycoproteins 1405:1417 arg1 glycoproteins 1405:1417 the salivary glycoproteins 1392:1417 the salivary glycoproteins of alcohol-dependent people 1392:1445 CONCLUSION Alterations in the glycosylation profiles in the salivary glycoproteins of alcohol-dependent people were found. 24085030 1 100 theme established 230:240 arg1 biomarker 262:270 well established and highly specific biomarker 225:270 well established and highly specific biomarker of sustained alcohol consumption 225:303 AIM Glycosylation of serum proteins is affected with prolonged heavy drinking, and carbohydrate deficient transferrin (CDT) is well established and highly specific biomarker of sustained alcohol consumption. 25038691 0 0 from changes 19:25 arg1 glycosylation 54:66 bovine milk lactoferrin glycosylation 30:66 bovine milk lactoferrin glycosylation using lectin microarrays 30:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 3 1 from cows 425:428 arg1 months 383:388 the first three months 367:388 the first three months of lactation from three individual cows 367:428 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 4 2 theme early 718:722 arg1 production 735:744 early stage milk production 718:744 early stage milk production 718:744 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 3 theme monosaccharide 589:602 arg1 analysis 604:611 accompanying monosaccharide analysis 576:611 accompanying monosaccharide analysis 576:611 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 4 theme diverse 785:791 arg1 structures 822:831 more diverse complex-type oligosaccharide structures 780:831 more diverse complex-type oligosaccharide structures 780:831 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 5 theme later 926:930 arg1 lactation 932:940 later lactation 926:940 later lactation 926:940 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 4 6 theme accompanying 576:587 arg1 analysis 604:611 accompanying monosaccharide analysis 576:611 accompanying monosaccharide analysis 576:611 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 7 theme profile 505:511 arg1 differences 513:523 Substantial profile differences 493:523 Substantial profile differences between early and late lactation 493:556 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 8 attach present 838:844 arg1 bLF 849:851 bLF 849:851 bLF 849:851 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 8 attach present 838:844 arg2 structures 822:831 more diverse complex-type oligosaccharide structures 780:831 more diverse complex-type oligosaccharide structures 780:831 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 4 9 theme N-glycolylneuraminic 672:691 arg1 acid 666:669 the non-human sialic acid 645:669 the non-human sialic acid 645:669 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 9 theme N-glycolylneuraminic 672:691 arg1 acid 693:696 N-glycolylneuraminic acid 672:696 N-glycolylneuraminic acid 672:696 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 3 10 theme time 348:351 arg1 points 353:358 13 time points 345:358 13 time points 345:358 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 6 11 contain have 1040:1043 arg2 functionality 1055:1067 different functionality 1045:1067 different functionality 1045:1067 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 6 11 contain have 1040:1043 arg1 these 1030:1034 these 1030:1034 these 1030:1034 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 2 12 theme structural 276:285 arg1 changes 287:293 bLF's oligosaccharide structural changes 254:293 bLF's oligosaccharide structural changes over the course of lactation 254:322 However, little is known about bLF's oligosaccharide structural changes over the course of lactation. 25038691 1 13 gly glycoprotein 110:121 arg1 glycoprotein 110:121 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 3 14 theme lectin 473:478 arg1 microarrays 480:490 lectin microarrays 473:490 lectin microarrays 473:490 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 2 15 theme oligosaccharide 260:274 arg1 changes 287:293 bLF's oligosaccharide structural changes 254:293 bLF's oligosaccharide structural changes over the course of lactation 254:322 However, little is known about bLF's oligosaccharide structural changes over the course of lactation. 25038691 5 16 theme oligomannose 897:908 arg1 glycans 915:921 oligomannose type glycans 897:921 oligomannose type glycans 897:921 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 3 17 theme individual 414:423 arg1 cows 425:428 three individual cows 408:428 three individual cows 408:428 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 1 18 contain has 148:150 arg2 variety 154:160 a variety 152:160 a variety of biological activities related to its constituent glycans 152:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 18 contain has 148:150 arg1 bLF 143:145 bLF 143:145 bLF 143:145 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 18 contain has 148:150 arg1 lactoferrin 130:140 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 18 contain has 148:150 arg2 activities 176:185 biological activities 165:185 biological activities related to its constituent glycans 165:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 5 19 theme complex-type 793:804 arg1 structures 822:831 more diverse complex-type oligosaccharide structures 780:831 more diverse complex-type oligosaccharide structures 780:831 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 1 20 theme biological 165:174 arg1 activities 176:185 biological activities 165:185 biological activities related to its constituent glycans 165:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 0 21 theme temporal 10:17 arg1 changes 19:25 temporal changes 10:25 temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays 10:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 6 22 from differences 947:957 arg1 glycoprofiles 966:978 the glycoprofiles 962:978 the glycoprofiles of bLF from colostrum to mature milk 962:1015 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 1 23 theme activities 176:185 arg1 activities 176:185 biological activities 165:185 biological activities related to its constituent glycans 165:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 23 theme activities 176:185 arg1 variety 154:160 a variety 152:160 a variety of biological activities related to its constituent glycans 152:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 3 24 theme first 371:375 arg1 months 383:388 the first three months 367:388 the first three months of lactation from three individual cows 367:428 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 6 25 theme mature 1005:1010 arg1 milk 1012:1015 mature milk 1005:1015 mature milk 1005:1015 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 1 26 theme bovine 98:103 arg1 bLF 143:145 bLF 143:145 bLF 143:145 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 26 theme bovine 98:103 arg1 lactoferrin 130:140 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 27 theme related 187:193 arg1 activities 176:185 biological activities 165:185 biological activities related to its constituent glycans 165:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 6 28 from colostrum 992:1000 arg1 bLF 983:985 bLF 983:985 bLF from colostrum to mature milk 983:1015 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 6 28 from colostrum 992:1000 arg1 glycoprofiles 966:978 the glycoprofiles 962:978 the glycoprofiles of bLF from colostrum to mature milk 962:1015 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 1 29 theme milk 105:108 arg1 bLF 143:145 bLF 143:145 bLF 143:145 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 29 theme milk 105:108 arg1 lactoferrin 130:140 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 4 30 theme non-human 649:657 arg1 acid 666:669 the non-human sialic acid 645:669 the non-human sialic acid 645:669 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 30 theme non-human 649:657 arg1 acid 693:696 N-glycolylneuraminic acid 672:696 N-glycolylneuraminic acid 672:696 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 1 31 theme glycoprotein 110:121 arg1 bLF 143:145 bLF 143:145 bLF 143:145 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 31 theme glycoprotein 110:121 arg1 lactoferrin 130:140 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 0 32 theme milk 37:40 arg1 glycosylation 54:66 bovine milk lactoferrin glycosylation 30:66 bovine milk lactoferrin glycosylation using lectin microarrays 30:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 4 33 theme acid 666:669 arg1 occurrence 631:640 the occurrence 627:640 the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, 627:697 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 33 theme acid 666:669 arg1 greater 703:709 greater 703:709 greater 703:709 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 34 theme type 910:913 arg1 glycans 915:921 oligomannose type glycans 897:921 oligomannose type glycans 897:921 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 1 35 theme constituent 202:212 arg1 glycans 214:220 its constituent glycans 198:220 its constituent glycans 198:220 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 0 36 theme bovine 30:35 arg1 glycosylation 54:66 bovine milk lactoferrin glycosylation 30:66 bovine milk lactoferrin glycosylation using lectin microarrays 30:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 4 37 theme early 533:537 arg1 lactation 548:556 early and late lactation 533:556 early and late lactation 533:556 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 3 38 theme lactation 393:401 arg1 months 383:388 the first three months 367:388 the first three months of lactation from three individual cows 367:428 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 3 39 theme glycosylation 434:446 arg1 changes 448:454 glycosylation changes 434:454 glycosylation changes 434:454 BLF was isolated at 13 time points during the first three months of lactation from three individual cows and glycosylation changes were profiled by lectin microarrays. 25038691 6 40 theme bLF 983:985 arg1 glycoprofiles 966:978 the glycoprofiles 962:978 the glycoprofiles of bLF from colostrum to mature milk 962:1015 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 6 41 from glycoprofiles 966:978 arg1 colostrum 992:1000 colostrum 992:1000 colostrum 992:1000 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 1 42 theme bovine 123:128 arg1 bLF 143:145 bLF 143:145 bLF 143:145 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 1 42 theme bovine 123:128 arg1 lactoferrin 130:140 The bovine milk glycoprotein bovine lactoferrin 94:140 The bovine milk glycoprotein bovine lactoferrin (bLF) 94:146 The bovine milk glycoprotein bovine lactoferrin (bLF) has a variety of biological activities related to its constituent glycans. 25038691 0 43 theme lactoferrin 42:52 arg1 glycosylation 54:66 bovine milk lactoferrin glycosylation 30:66 bovine milk lactoferrin glycosylation using lectin microarrays 30:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 4 44 theme late 543:546 arg1 lactation 548:556 early and late lactation 533:556 early and late lactation 533:556 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 45 theme stage 724:728 arg1 production 735:744 early stage milk production 718:744 early stage milk production 718:744 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 46 theme milk 730:733 arg1 production 735:744 early stage milk production 718:744 early stage milk production 718:744 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 47 located present 838:844 arg1 bLF 849:851 bLF 849:851 bLF 849:851 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 47 located present 838:844 arg2 structures 822:831 more diverse complex-type oligosaccharide structures 780:831 more diverse complex-type oligosaccharide structures 780:831 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 48 with present 838:844 arg1 abundance 884:892 an abundance 881:892 an abundance of oligomannose type glycans in later lactation 881:940 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 2 49 theme lactation 314:322 arg1 course 304:309 the course 300:309 the course of lactation 300:322 However, little is known about bLF's oligosaccharide structural changes over the course of lactation. 25038691 6 50 theme different 1045:1053 arg1 functionality 1055:1067 different functionality 1045:1067 different functionality 1045:1067 The differences in the glycoprofiles of bLF from colostrum to mature milk suggest that these may have different functionality in vivo. 25038691 0 51 theme lectin 74:79 arg1 microarrays 81:91 lectin microarrays 74:91 lectin microarrays 74:91 Profiling temporal changes in bovine milk lactoferrin glycosylation using lectin microarrays. 25038691 4 52 theme sialic 659:664 arg1 acid 666:669 the non-human sialic acid 645:669 the non-human sialic acid 645:669 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 4 52 theme sialic 659:664 arg1 acid 693:696 N-glycolylneuraminic acid 672:696 N-glycolylneuraminic acid 672:696 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 53 theme early 860:864 arg1 lactation 866:874 early lactation 860:874 early lactation 860:874 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 54 theme glycans 915:921 arg1 abundance 884:892 an abundance 881:892 an abundance of oligomannose type glycans in later lactation 881:940 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 4 55 theme Substantial 493:503 arg1 differences 513:523 Substantial profile differences 493:523 Substantial profile differences between early and late lactation 493:556 Substantial profile differences between early and late lactation were observed and accompanying monosaccharide analysis revealed that the occurrence of the non-human sialic acid, N-glycolylneuraminic acid, was greater during early stage milk production. 25038691 5 56 theme oligosaccharide 806:820 arg1 structures 822:831 more diverse complex-type oligosaccharide structures 780:831 more diverse complex-type oligosaccharide structures 780:831 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25038691 5 57 from abundance 884:892 arg1 lactation 932:940 later lactation 926:940 later lactation 926:940 Overall, the data suggested that more diverse complex-type oligosaccharide structures were present on bLF during early lactation with an abundance of oligomannose type glycans in later lactation. 25799047 5 0 theme protein 1236:1242 arg1 quantities 1210:1219 significant quantities 1198:1219 significant quantities of full-length protein 1198:1242 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 8 1 theme protein 1682:1688 arg1 transport 1690:1698 either protein transport 1675:1698 either protein transport 1675:1698 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 2 2 theme different 338:346 arg1 species 348:354 different species 338:354 different species 338:354 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 10 3 theme important 2137:2145 arg1 enzyme 2147:2152 this important enzyme 2132:2152 this important enzyme in biomass deconstruction 2132:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 1 4 theme cellulolytic 196:207 arg1 microbes 209:216 the most thermophilic cellulolytic microbes 174:216 the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment 174:307 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 1 4 theme cellulolytic 196:207 arg1 Members 118:124 Members 118:124 Members of the bacterial genus Caldicellulosiruptor 118:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 7 5 theme signal 1404:1409 arg1 sequence 1411:1418 its signal sequence 1400:1418 its signal sequence 1400:1418 Expression of CelA protein with and without its signal sequence allowed comparison of protein retained intracellularly to protein transported extracellularly. 25799047 8 6 theme protein 1791:1797 arg1 export 1799:1804 protein export 1791:1804 protein export 1791:1804 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 7 7 theme protein 1375:1381 arg1 Expression 1356:1365 Expression 1356:1365 Expression of CelA protein with and without its signal sequence 1356:1418 Expression of CelA protein with and without its signal sequence allowed comparison of protein retained intracellularly to protein transported extracellularly. 25799047 10 8 from enzyme 2147:2152 arg1 deconstruction 2165:2178 biomass deconstruction 2157:2178 biomass deconstruction 2157:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 9 9 theme mechanism 1969:1977 arg1 study 1956:1960 the study 1952:1960 the study of the mechanism of protein glycosylation in this thermophile 1952:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 5 10 theme new 1116:1118 arg1 vector 1131:1136 a new expression vector 1114:1136 a new expression vector for C. bescii 1114:1150 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 5 11 from role 1067:1070 arg1 deconstruction 1083:1096 biomass deconstruction 1075:1096 biomass deconstruction 1075:1096 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 4 12 theme complex 996:1002 arg1 biomass 1004:1010 complex biomass 996:1010 complex biomass 996:1010 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 9 13 theme protein 1982:1988 arg1 glycosylation 1990:2002 protein glycosylation 1982:2002 protein glycosylation in this thermophile 1982:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 9 14 dep mechanism 1811:1819 arg1 The 1807:1809 The 1807:1809 The 1807:1809 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 2 15 contain contains 451:458 arg2 family 487:492 a glycoside hydrolase family 9 endoglucanase 465:508 a glycoside hydrolase family 9 endoglucanase 465:508 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 15 contain contains 451:458 arg2 family 536:541 a glycoside hydrolase family 48 514:544 a glycoside hydrolase family 48 514:544 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 15 contain contains 451:458 arg1 CelA 439:442 the multimodular cellulase CelA 412:442 the multimodular cellulase CelA 412:442 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 16 theme linker 651:656 arg1 peptides 658:665 linker peptides 651:665 linker peptides 651:665 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 7 17 theme CelA 1370:1373 arg1 protein 1375:1381 CelA protein 1370:1381 CelA protein 1370:1381 Expression of CelA protein with and without its signal sequence allowed comparison of protein retained intracellularly to protein transported extracellularly. 25799047 2 18 from synergistic 571:581 arg1 activity 592:599 their activity 586:599 their activity 586:599 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 3 19 theme surface 709:715 arg1 ablation 717:724 the cellulose surface ablation 695:724 the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities 695:799 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 1 20 theme genus 143:147 arg1 Caldicellulosiruptor 149:168 the bacterial genus Caldicellulosiruptor 129:168 the bacterial genus Caldicellulosiruptor 129:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 3 21 theme cellulase 867:875 arg1 activity 877:884 cellulase activity 867:884 cellulase activity 867:884 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 9 22 gly glycosylation 1990:2002 arg1 thermophile 2012:2022 this thermophile 2007:2022 this thermophile 2007:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 4 23 contain had 945:947 arg1 deletion 915:922 a deletion 913:922 a deletion of celA in C. bescii 913:943 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 4 23 contain had 945:947 arg2 effect 963:968 a significant effect 949:968 a significant effect 949:968 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 0 24 theme extracellular 79:91 arg1 protein 93:99 the extracellular protein 75:99 the extracellular protein 75:99 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 1 25 theme most 178:181 arg1 microbes 209:216 the most thermophilic cellulolytic microbes 174:216 the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment 174:307 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 1 25 theme most 178:181 arg1 Members 118:124 Members 118:124 Members of the bacterial genus Caldicellulosiruptor 118:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 9 26 theme glycosylation 1841:1853 arg1 role 1825:1828 role 1825:1828 role 1825:1828 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 9 26 theme glycosylation 1841:1853 arg1 mechanism 1811:1819 mechanism 1811:1819 mechanism 1811:1819 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 10 27 from structure 2106:2114 arg1 study 2048:2052 the study 2044:2052 the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction 2044:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 27 from structure 2106:2114 arg1 deconstruction 2165:2178 biomass deconstruction 2157:2178 biomass deconstruction 2157:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 0 28 theme Homologous 0:9 arg1 expression 11:20 Homologous expression 0:20 Homologous expression of the Caldicellulosiruptor bescii CelA 0:60 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 0 29 gly glycosylated 104:115 arg1 protein 93:99 the extracellular protein 75:99 the extracellular protein 75:99 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 8 30 theme extracellular 1577:1589 arg1 protein 1596:1602 the extracellular CelA protein 1573:1602 the extracellular CelA protein 1573:1602 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 10 31 from function 2120:2127 arg1 study 2048:2052 the study 2044:2052 the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction 2044:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 31 from function 2120:2127 arg1 deconstruction 2165:2178 biomass deconstruction 2157:2178 biomass deconstruction 2157:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 9 32 from thermophile 2012:2022 arg1 mechanism 1969:1977 the mechanism 1965:1977 the mechanism of protein glycosylation in this thermophile 1965:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 2 33 theme glycoside 467:475 arg1 family 487:492 a glycoside hydrolase family 9 endoglucanase 465:508 a glycoside hydrolase family 9 endoglucanase 465:508 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 0 34 theme Caldicellulosiruptor 29:48 arg1 CelA 57:60 the Caldicellulosiruptor bescii CelA 25:60 the Caldicellulosiruptor bescii CelA 25:60 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 5 35 theme first 1175:1179 arg1 time 1181:1184 the first time 1171:1184 the first time 1171:1184 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 10 36 from study 2048:2052 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 36 from study 2048:2052 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 2 37 theme hydrolase 526:534 arg1 family 536:541 a glycoside hydrolase family 48 514:544 a glycoside hydrolase family 48 514:544 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 38 dep family 487:492 arg1 exoglucanase 546:557 exoglucanase 546:557 exoglucanase 546:557 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 38 dep family 487:492 arg1 endoglucanase 496:508 endoglucanase 496:508 a glycoside hydrolase family 9 endoglucanase 465:508 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 0 39 theme CelA 57:60 arg1 expression 11:20 Homologous expression 0:20 Homologous expression of the Caldicellulosiruptor bescii CelA 0:60 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 10 40 from glycosylation 2057:2069 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 40 from glycosylation 2057:2069 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 2 41 theme cellulolytic 314:325 arg1 ability 327:333 The cellulolytic ability 310:333 The cellulolytic ability of different species 310:354 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 1 42 theme lignocellulosic 251:265 arg1 biomass 267:273 lignocellulosic biomass 251:273 lignocellulosic biomass 251:273 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 5 43 theme CelA 1054:1057 arg1 structure 1028:1036 structure 1028:1036 structure 1028:1036 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 5 43 theme CelA 1054:1057 arg1 role 1067:1070 its role 1063:1070 its role in biomass deconstruction 1063:1096 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 5 43 theme CelA 1054:1057 arg1 function 1042:1049 function 1042:1049 function 1042:1049 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 3 44 theme cellulase 748:756 arg1 processivity 758:769 its general cellulase processivity 736:769 its general cellulase processivity 736:769 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 2 45 theme CelA 439:442 arg1 presence 400:407 the presence 396:407 the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides 396:665 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 5 46 theme native 1259:1264 arg1 host 1266:1269 the native host 1255:1269 the native host 1255:1269 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 6 47 theme Histidine 1302:1310 arg1 tag 1312:1314 a Histidine tag 1300:1314 a Histidine tag 1300:1314 The protein, which contains a Histidine tag, was active and excreted from the cell. 25799047 2 48 theme multimodular 416:427 arg1 CelA 439:442 the multimodular cellulase CelA 412:442 the multimodular cellulase CelA 412:442 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 5 49 from function 1042:1049 arg1 deconstruction 1083:1096 biomass deconstruction 1075:1096 biomass deconstruction 1075:1096 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 5 50 theme full-length 1224:1234 arg1 protein 1236:1242 full-length protein 1224:1242 full-length protein 1224:1242 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 10 51 dep structure 2106:2114 arg1 the 2102:2104 the 2102:2104 the 2102:2104 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 52 gly glycosylation 2057:2069 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 52 gly glycosylation 2057:2069 arg1 CelA 2074:2077 CelA itself 2074:2084 CelA itself 2074:2084 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 52 gly glycosylation 2057:2069 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 2 53 theme species 348:354 arg1 ability 327:333 The cellulolytic ability 310:333 The cellulolytic ability of different species 310:354 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 5 54 from structure 1028:1036 arg1 deconstruction 1083:1096 biomass deconstruction 1075:1096 biomass deconstruction 1075:1096 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 7 55 theme protein 1442:1448 arg1 comparison 1428:1437 comparison 1428:1437 comparison of protein retained intracellularly to protein transported extracellularly 1428:1512 Expression of CelA protein with and without its signal sequence allowed comparison of protein retained intracellularly to protein transported extracellularly. 25799047 10 56 theme enzyme 2147:2152 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 56 theme enzyme 2147:2152 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 1 57 theme thermophilic 183:194 arg1 microbes 209:216 the most thermophilic cellulolytic microbes 174:216 the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment 174:307 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 1 57 theme thermophilic 183:194 arg1 Members 118:124 Members 118:124 Members of the bacterial genus Caldicellulosiruptor 118:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 4 58 theme celA 927:930 arg1 deletion 915:922 a deletion 913:922 a deletion of celA in C. bescii 913:943 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 8 59 gly glycosylation 1761:1773 arg1 protein 1791:1797 protein export 1791:1804 protein export 1791:1804 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 10 60 theme biomass 2157:2163 arg1 deconstruction 2165:2178 biomass deconstruction 2157:2178 biomass deconstruction 2157:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 3 61 theme substrate 825:833 arg1 surface 810:816 the surface 806:816 the surface of the substrate 806:833 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 5 62 theme expression 1120:1129 arg1 vector 1131:1136 a new expression vector 1114:1136 a new expression vector for C. bescii 1114:1150 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 9 63 theme glycosylation 1990:2002 arg1 mechanism 1969:1977 the mechanism 1965:1977 the mechanism of protein glycosylation in this thermophile 1965:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 4 64 theme significant 951:961 arg1 effect 963:968 a significant effect 949:968 a significant effect 949:968 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 9 65 from mechanism 1969:1977 arg1 thermophile 2012:2022 this thermophile 2007:2022 this thermophile 2007:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 3 66 theme novel 848:852 arg1 paradigm 854:861 a novel paradigm 846:861 a novel paradigm for cellulase activity 846:884 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 3 67 theme cellulose 699:707 arg1 ablation 717:724 the cellulose surface ablation 695:724 the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities 695:799 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 1 68 theme bacterial 133:141 arg1 Caldicellulosiruptor 149:168 the bacterial genus Caldicellulosiruptor 129:168 the bacterial genus Caldicellulosiruptor 129:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 2 69 theme cellulose-binding 621:637 arg1 domains 639:645 three cellulose-binding domains 615:645 three cellulose-binding domains 615:645 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 2 70 from activity 592:599 arg1 synergistic 571:581 synergistic 571:581 synergistic 571:581 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 5 71 dep structure 1028:1036 arg1 the 1024:1026 the 1024:1026 the 1024:1026 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 1 72 theme Caldicellulosiruptor 149:168 arg1 microbes 209:216 the most thermophilic cellulolytic microbes 174:216 the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment 174:307 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 1 72 theme Caldicellulosiruptor 149:168 arg1 Members 118:124 Members 118:124 Members of the bacterial genus Caldicellulosiruptor 118:168 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 25799047 8 73 theme protein 1527:1533 arg1 Analysis 1515:1522 Analysis 1515:1522 Analysis of protein in culture supernatants 1515:1557 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 8 74 theme culture 1538:1544 arg1 supernatants 1546:1557 culture supernatants 1538:1557 culture supernatants 1538:1557 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 10 75 theme CelA 2074:2077 arg1 role 2094:2097 its role 2090:2097 its role in the structure and function of this important enzyme in biomass deconstruction 2090:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 75 theme CelA 2074:2077 arg1 glycosylation 2057:2069 glycosylation 2057:2069 glycosylation of CelA itself 2057:2084 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 8 76 from Analysis 1515:1522 arg1 supernatants 1546:1557 culture supernatants 1538:1557 culture supernatants 1538:1557 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 9 77 theme protein 1833:1839 arg1 glycosylation 1841:1853 protein glycosylation 1833:1853 protein glycosylation 1833:1853 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 10 78 theme glycosylation 2057:2069 arg1 study 2048:2052 the study 2044:2052 the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction 2044:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 6 79 contain contains 1291:1298 arg1 protein 1276:1282 The protein 1272:1282 The protein 1272:1282 The protein, which contains a Histidine tag, was active and excreted from the cell. 25799047 6 79 contain contains 1291:1298 arg2 tag 1312:1314 a Histidine tag 1300:1314 a Histidine tag 1300:1314 The protein, which contains a Histidine tag, was active and excreted from the cell. 25799047 10 80 from role 2094:2097 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 80 from role 2094:2097 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 4 81 from deletion 915:922 arg1 bescii 938:943 C. bescii 935:943 C. bescii 935:943 We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. 25799047 8 82 gly glycosylated 1607:1618 arg1 protein 1596:1602 the extracellular CelA protein 1573:1602 the extracellular CelA protein 1573:1602 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 8 83 theme CelA 1591:1594 arg1 protein 1596:1602 the extracellular CelA protein 1573:1602 the extracellular CelA protein 1573:1602 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 5 84 theme significant 1198:1208 arg1 quantities 1210:1219 significant quantities 1198:1219 significant quantities of full-length protein 1198:1242 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 5 85 dep able 1161:1164 arg1 express 1190:1196 express 1190:1196 to express significant quantities of full-length protein in vivo in the native host 1187:1269 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 2 86 theme hydrolase 477:485 arg1 family 487:492 a glycoside hydrolase family 9 endoglucanase 465:508 a glycoside hydrolase family 9 endoglucanase 465:508 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 0 87 theme bescii 50:55 arg1 CelA 57:60 the Caldicellulosiruptor bescii CelA 25:60 the Caldicellulosiruptor bescii CelA 25:60 Homologous expression of the Caldicellulosiruptor bescii CelA reveals that the extracellular protein is glycosylated. 25799047 9 88 from glycosylation 1990:2002 arg1 thermophile 2012:2022 this thermophile 2007:2022 this thermophile 2007:2022 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 5 89 theme biomass 1075:1081 arg1 deconstruction 1083:1096 biomass deconstruction 1075:1096 biomass deconstruction 1075:1096 To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. 25799047 2 90 theme glycoside 516:524 arg1 family 536:541 a glycoside hydrolase family 48 514:544 a glycoside hydrolase family 48 514:544 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 8 91 theme intracellular 1632:1644 arg1 CelA 1646:1649 the intracellular CelA 1628:1649 the intracellular CelA 1628:1649 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 9 92 from role 1825:1828 arg1 bacteria 1858:1865 bacteria 1858:1865 bacteria 1858:1865 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 10 93 theme role 2094:2097 arg1 study 2048:2052 the study 2044:2052 the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction 2044:2178 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 3 94 theme general 740:746 arg1 processivity 758:769 its general cellulase processivity 736:769 its general cellulase processivity 736:769 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 9 95 from mechanism 1811:1819 arg1 bacteria 1858:1865 bacteria 1858:1865 bacteria 1858:1865 The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. 25799047 3 96 theme excavates 782:790 arg1 cavities 792:799 excavates cavities 782:799 the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities 695:799 This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm for cellulase activity. 25799047 10 97 from deconstruction 2165:2178 arg1 structure 2106:2114 structure 2106:2114 structure 2106:2114 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 10 97 from deconstruction 2165:2178 arg1 function 2120:2127 function 2120:2127 function 2120:2127 It will also allow the study of glycosylation of CelA itself and its role in the structure and function of this important enzyme in biomass deconstruction. 25799047 2 98 theme cellulase 429:437 arg1 CelA 439:442 the multimodular cellulase CelA 412:442 the multimodular cellulase CelA 412:442 The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. 25799047 8 99 theme post-translational 1721:1738 arg1 modification 1740:1751 this post-translational modification 1716:1751 this post-translational modification 1716:1751 Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. 25799047 1 100 theme conventional 283:294 arg1 pretreatment 296:307 conventional pretreatment 283:307 conventional pretreatment 283:307 Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. 24024472 3 0 theme GP2013 365:370 arg1 bioactivities 348:360 bioactivities 348:360 bioactivities 348:360 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 3 0 theme GP2013 365:370 arg1 modifications 330:342 Post-translational modifications 311:342 Post-translational modifications 311:342 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 5 1 theme drug 792:795 arg1 product 797:803 clinical scale drug product 777:803 clinical scale drug product 777:803 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 9 2 dep GP2013 1277:1282 arg1 rituximab 1299:1307 rituximab 1299:1307 rituximab 1299:1307 This preclinical comparability exercise confirms that GP2013 and originator rituximab are pharmacologically similar. 24024472 1 3 theme similar 186:192 arg1 product 194:200 a similar product 184:200 a similar product to the originator 184:218 Biosimilar development involves a target-directed iterative process to ensure a similar product to the originator. 24024472 0 4 theme GP2013 98:103 arg1 development 16:26 Target-directed development 0:26 Target-directed development 0:26 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 0 4 theme GP2013 98:103 arg1 characterization 44:59 preclinical characterization 32:59 preclinical characterization 32:59 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 5 5 dep GP2013 701:706 arg1 rituximab 723:731 rituximab 723:731 rituximab 723:731 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 7 6 theme sensitive 1076:1084 arg1 levels 1107:1112 sensitive sub-therapeutic dose levels 1076:1112 sensitive sub-therapeutic dose levels 1076:1112 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 6 7 theme killer 962:967 arg1 cells 974:978 freshly purified human natural killer (NK) cells 931:978 freshly purified human natural killer (NK) cells 931:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 8 8 from comparable 1189:1198 arg1 monkeys 1214:1220 cynomolgus monkeys 1203:1220 cynomolgus monkeys 1203:1220 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 4 9 theme different 549:557 arg1 patterns 573:580 different glycosylation patterns 549:580 different glycosylation patterns 549:580 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 6 10 theme lymphoma 905:912 arg1 lines 919:923 two lymphoma cell lines 901:923 two lymphoma cell lines using freshly purified human natural killer (NK) cells 901:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 4 11 theme cellular 493:500 arg1 ADCC 516:519 ADCC 516:519 ADCC 516:519 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 4 11 theme cellular 493:500 arg1 cytotoxicity 502:513 Antibody-dependent cellular cytotoxicity 474:513 Antibody-dependent cellular cytotoxicity (ADCC) 474:520 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 9 12 theme preclinical 1228:1238 arg1 exercise 1254:1261 This preclinical comparability exercise 1223:1261 This preclinical comparability exercise 1223:1261 This preclinical comparability exercise confirms that GP2013 and originator rituximab are pharmacologically similar. 24024472 4 13 used used 526:529 arg2 cytotoxicity 502:513 Antibody-dependent cellular cytotoxicity 474:513 Antibody-dependent cellular cytotoxicity (ADCC) 474:520 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 4 13 used used 526:529 arg2 ADCC 516:519 ADCC 516:519 ADCC 516:519 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 8 14 theme CD20 1154:1157 arg1 Pharmacokinetics 1115:1130 Pharmacokinetics 1115:1130 Pharmacokinetics 1115:1130 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 8 14 theme CD20 1154:1157 arg1 depletion 1164:1172 CD20 cell depletion 1154:1172 CD20 cell depletion 1154:1172 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 4 15 theme Antibody-dependent 474:491 arg1 ADCC 516:519 ADCC 516:519 ADCC 516:519 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 4 15 theme Antibody-dependent 474:491 arg1 cytotoxicity 502:513 Antibody-dependent cellular cytotoxicity 474:513 Antibody-dependent cellular cytotoxicity (ADCC) 474:520 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 7 16 theme characterized 1027:1039 arg1 models 1057:1062 two well characterized mouse xenograft models 1018:1062 two well characterized mouse xenograft models 1018:1062 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 8 17 theme cynomolgus 1203:1212 arg1 monkeys 1214:1220 cynomolgus monkeys 1203:1220 cynomolgus monkeys 1203:1220 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 6 18 theme purified 939:946 arg1 cells 974:978 freshly purified human natural killer (NK) cells 931:978 freshly purified human natural killer (NK) cells 931:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 0 19 theme Target-directed 0:14 arg1 development 16:26 Target-directed development 0:26 Target-directed development 0:26 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 7 20 theme In 981:982 arg1 efficacy 989:996 In vivo efficacy 981:996 In vivo efficacy 981:996 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 3 21 theme similar 440:446 arg1 profiles 464:471 similar pharmacological profiles 440:471 similar pharmacological profiles 440:471 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 4 22 theme glycosylation 559:571 arg1 patterns 573:580 different glycosylation patterns 549:580 different glycosylation patterns 549:580 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 5 23 theme clinical 777:784 arg1 product 797:803 clinical scale drug product 777:803 clinical scale drug product 777:803 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 0 24 theme rituximab 88:96 arg1 GP2013 98:103 the proposed biosimilar rituximab GP2013 64:103 the proposed biosimilar rituximab GP2013 64:103 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 1 25 theme Biosimilar 106:115 arg1 development 117:127 Biosimilar development 106:127 Biosimilar development 106:127 Biosimilar development involves a target-directed iterative process to ensure a similar product to the originator. 24024472 7 26 dep In 981:982 arg1 vivo 984:987 vivo 984:987 vivo 984:987 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 3 27 theme pharmacological 448:462 arg1 profiles 464:471 similar pharmacological profiles 440:471 similar pharmacological profiles 440:471 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 5 28 theme scale 786:790 arg1 product 797:803 clinical scale drug product 777:803 clinical scale drug product 777:803 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 0 29 theme preclinical 32:42 arg1 characterization 44:59 preclinical characterization 32:59 preclinical characterization 32:59 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 7 30 theme sub-therapeutic 1086:1100 arg1 levels 1107:1112 sensitive sub-therapeutic dose levels 1076:1112 sensitive sub-therapeutic dose levels 1076:1112 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 5 31 theme preclinical 751:761 arg1 studies 763:769 preclinical studies 751:769 preclinical studies using clinical scale drug product 751:803 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 6 32 theme ADCC 823:826 arg1 potency 828:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 3 33 theme Post-translational 311:328 arg1 modifications 330:342 Post-translational modifications 311:342 Post-translational modifications 311:342 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 7 34 theme dose 1102:1105 arg1 levels 1107:1112 sensitive sub-therapeutic dose levels 1076:1112 sensitive sub-therapeutic dose levels 1076:1112 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 9 35 theme comparability 1240:1252 arg1 exercise 1254:1261 This preclinical comparability exercise 1223:1261 This preclinical comparability exercise 1223:1261 This preclinical comparability exercise confirms that GP2013 and originator rituximab are pharmacologically similar. 24024472 6 36 theme in 814:815 arg1 potency 828:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 7 37 theme xenograft 1047:1055 arg1 models 1057:1062 two well characterized mouse xenograft models 1018:1062 two well characterized mouse xenograft models 1018:1062 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 6 38 theme cell 914:917 arg1 lines 919:923 two lymphoma cell lines 901:923 two lymphoma cell lines using freshly purified human natural killer (NK) cells 901:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 0 39 theme proposed 68:75 arg1 GP2013 98:103 the proposed biosimilar rituximab GP2013 64:103 the proposed biosimilar rituximab GP2013 64:103 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 6 40 theme natural 954:960 arg1 NK 970:971 NK 970:971 NK 970:971 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 6 40 theme natural 954:960 arg1 killer 962:967 human natural killer 948:967 freshly purified human natural killer (NK) cells 931:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 0 41 theme biosimilar 77:86 arg1 GP2013 98:103 the proposed biosimilar rituximab GP2013 64:103 the proposed biosimilar rituximab GP2013 64:103 Target-directed development and preclinical characterization of the proposed biosimilar rituximab GP2013. 24024472 5 42 theme Pharmacological 663:677 arg1 comparability 679:691 Pharmacological comparability 663:691 Pharmacological comparability between GP2013 and originator rituximab 663:731 Pharmacological comparability between GP2013 and originator rituximab were confirmed in preclinical studies using clinical scale drug product. 24024472 1 43 theme target-directed 140:154 arg1 process 166:172 a target-directed iterative process 138:172 a target-directed iterative process to ensure a similar product to the originator 138:218 Biosimilar development involves a target-directed iterative process to ensure a similar product to the originator. 24024472 8 44 from monkeys 1214:1220 arg1 comparable 1189:1198 comparable 1189:1198 comparable 1189:1198 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 2 45 theme preclinical 240:250 arg1 development 252:262 the preclinical development 236:262 the preclinical development of the proposed biosimilar rituximab (GP2013) 236:308 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 24024472 1 46 theme iterative 156:164 arg1 process 166:172 a target-directed iterative process 138:172 a target-directed iterative process to ensure a similar product to the originator 138:218 Biosimilar development involves a target-directed iterative process to ensure a similar product to the originator. 24024472 6 47 theme Similar 806:812 arg1 potency 828:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency 806:834 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 4 48 theme process 642:648 arg1 development 650:660 process development 642:660 process development 642:660 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 8 49 theme cell 1159:1162 arg1 Pharmacokinetics 1115:1130 Pharmacokinetics 1115:1130 Pharmacokinetics 1115:1130 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 8 49 theme cell 1159:1162 arg1 depletion 1164:1172 CD20 cell depletion 1154:1172 CD20 cell depletion 1154:1172 Pharmacokinetics and pharmacodynamics (CD20 cell depletion) were likewise comparable in cynomolgus monkeys. 24024472 6 50 dep in 814:815 arg1 vitro 817:821 vitro 817:821 vitro 817:821 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 2 51 theme rituximab 291:299 arg1 development 252:262 the preclinical development 236:262 the preclinical development of the proposed biosimilar rituximab (GP2013) 236:308 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 24024472 6 52 theme human 948:952 arg1 NK 970:971 NK 970:971 NK 970:971 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 6 52 theme human 948:952 arg1 killer 962:967 human natural killer 948:967 freshly purified human natural killer (NK) cells 931:978 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 3 53 theme originator 379:388 arg1 rituximab 390:398 originator rituximab 379:398 originator rituximab 379:398 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 2 54 theme biosimilar 280:289 arg1 GP2013 302:307 GP2013 302:307 GP2013 302:307 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 24024472 2 54 theme biosimilar 280:289 arg1 rituximab 291:299 the proposed biosimilar rituximab 267:299 the proposed biosimilar rituximab (GP2013) 267:308 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 24024472 6 55 theme dose-response 872:884 arg1 manner 886:891 a dose-response manner 870:891 a dose-response manner 870:891 Similar in vitro ADCC potency was demonstrated when compared in a dose-response manner against two lymphoma cell lines using freshly purified human natural killer (NK) cells. 24024472 7 56 theme mouse 1041:1045 arg1 models 1057:1062 two well characterized mouse xenograft models 1018:1062 two well characterized mouse xenograft models 1018:1062 In vivo efficacy was demonstrated in two well characterized mouse xenograft models, testing at sensitive sub-therapeutic dose levels. 24024472 3 57 theme rituximab 390:398 arg1 bioactivities 348:360 bioactivities 348:360 bioactivities 348:360 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 3 57 theme rituximab 390:398 arg1 modifications 330:342 Post-translational modifications 311:342 Post-translational modifications 311:342 Post-translational modifications and bioactivities of GP2013 versus originator rituximab were engineered and monitored to ensure similar pharmacological profiles. 24024472 4 58 theme structure-function 586:603 arg1 relationships 605:617 structure-function relationships 586:617 structure-function relationships 586:617 Antibody-dependent cellular cytotoxicity (ADCC) was used to illustrate how different glycosylation patterns and structure-function relationships were controlled during process development. 24024472 2 59 theme proposed 271:278 arg1 GP2013 302:307 GP2013 302:307 GP2013 302:307 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 24024472 2 59 theme proposed 271:278 arg1 rituximab 291:299 the proposed biosimilar rituximab 267:299 the proposed biosimilar rituximab (GP2013) 267:308 Here we report the preclinical development of the proposed biosimilar rituximab (GP2013). 28681696 4 0 theme healthy 996:1002 arg1 controls 1004:1011 healthy controls 996:1011 healthy controls 996:1011 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 8 1 theme cancer 2055:2060 arg1 diagnosis 2062:2070 epithelial ovarian cancer diagnosis 2036:2070 epithelial ovarian cancer diagnosis 2036:2070 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 3 2 theme tumor 525:529 arg1 tissues 531:537 tumor tissues 525:537 tumor tissues 525:537 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 7 3 theme high-mannose-type 1629:1645 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 6 4 theme complex 1412:1418 arg1 N-glycans 1420:1428 four high-mannose and three fucosylated neutral complex N-glycans 1364:1428 N-glycans 1420:1428 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 8 5 theme N-glycomic 1884:1893 arg1 changes 1895:1901 N-glycomic changes 1884:1901 N-glycomic changes 1884:1901 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 8 5 theme N-glycomic 1884:1893 arg1 indicator 1916:1924 a strong indicator 1907:1924 a strong indicator for epithelial ovarian cancer pathological grades 1907:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 4 6 theme epithelial 966:975 arg1 cancer 985:990 epithelial ovarian cancer 966:990 epithelial ovarian cancer 966:990 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 7 7 gly fucosylated 1704:1714 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 1 8 attach linked 179:184 arg2 changes 123:129 Aberrant changes 114:129 Aberrant changes of N-glycan modifications on proteins 114:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 1 8 attach linked 179:184 arg1 diseases 197:204 various diseases 189:204 various diseases including different cancers 189:232 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 1 8 attach linked 179:184 arg1 cancers 226:232 different cancers 216:232 different cancers 216:232 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 6 9 theme fucosylated 1392:1402 arg1 N-glycans 1420:1428 four high-mannose and three fucosylated neutral complex N-glycans 1364:1428 N-glycans 1420:1428 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 3 10 theme ovarian 557:563 arg1 cancer 565:570 epithelial ovarian cancer 546:570 epithelial ovarian cancer progression 546:582 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 1 11 theme various 189:195 arg1 diseases 197:204 various diseases 189:204 various diseases including different cancers 189:232 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 1 11 theme various 189:195 arg1 cancers 226:232 different cancers 216:232 different cancers 216:232 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 4 12 theme cancer 985:990 arg1 controls 1004:1011 healthy controls 996:1011 healthy controls 996:1011 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 4 12 theme cancer 985:990 arg1 grades 956:961 different pathological grades 933:961 different pathological grades of epithelial ovarian cancer 933:990 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 6 13 theme characteristic 1297:1310 arg1 analysis 1312:1319 characteristic analysis 1297:1319 characteristic analysis 1297:1319 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 4 14 theme ovarian 726:732 arg1 cancer 734:739 epithelial ovarian cancer 715:739 epithelial ovarian cancer 715:739 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 8 15 theme strong 1909:1914 arg1 changes 1895:1901 N-glycomic changes 1884:1901 N-glycomic changes 1884:1901 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 8 15 theme strong 1909:1914 arg1 indicator 1916:1924 a strong indicator 1907:1924 a strong indicator for epithelial ovarian cancer pathological grades 1907:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 8 16 theme ovarian 2047:2053 arg1 diagnosis 2062:2070 epithelial ovarian cancer diagnosis 2036:2070 epithelial ovarian cancer diagnosis 2036:2070 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 5 17 dep higher 1128:1133 arg1 accompanied 1212:1222 accompanied 1212:1222 accompanied by reduced levels of hybrid-type glycans 1212:1263 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 4 18 dep formalin-fixed 769:782 arg1 paraffin-embedded 784:800 paraffin-embedded 784:800 paraffin-embedded 784:800 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 5 19 theme healthy 1194:1200 arg1 controls 1202:1209 healthy controls 1194:1209 healthy controls 1194:1209 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 7 20 theme ovarian 1810:1816 arg1 grades 1825:1830 epithelial ovarian cancer grades 1799:1830 epithelial ovarian cancer grades 1799:1830 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 21 theme statistical 1534:1544 arg1 analysis 1546:1553 a statistical analysis 1532:1553 a statistical analysis of variance assay 1532:1571 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 6 22 theme ovarian 1475:1481 arg1 cancer 1483:1488 epithelial ovarian cancer 1464:1488 epithelial ovarian cancer 1464:1488 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 8 23 theme ovarian 1941:1947 arg1 grades 1969:1974 epithelial ovarian cancer pathological grades 1930:1974 epithelial ovarian cancer pathological grades 1930:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 5 24 located observed 1182:1189 arg1 controls 1202:1209 healthy controls 1194:1209 healthy controls 1194:1209 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 5 24 located observed 1182:1189 arg2 that 1177:1180 that 1177:1180 that 1177:1180 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 8 25 theme pathological 1956:1967 arg1 grades 1969:1974 epithelial ovarian cancer pathological grades 1930:1974 epithelial ovarian cancer pathological grades 1930:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 1 26 theme N-glycan 134:141 arg1 modifications 143:155 N-glycan modifications 134:155 N-glycan modifications on proteins 134:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 7 27 theme N-glycan 1773:1780 arg1 abundance 1782:1790 N-glycan abundance 1773:1790 N-glycan abundance 1773:1790 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 6 28 theme good 1441:1444 arg1 discrimination 1446:1459 good discrimination 1441:1459 good discrimination of epithelial ovarian cancer from healthy controls 1441:1510 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 2 29 from Changes 323:329 arg1 patterns 343:350 N-glycan patterns 334:350 N-glycan patterns 334:350 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 28681696 5 30 theme hybrid-type 1245:1255 arg1 glycans 1257:1263 hybrid-type glycans 1245:1263 hybrid-type glycans 1245:1263 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 0 31 theme ovarian 86:92 arg1 cancer 94:99 epithelial ovarian cancer 75:99 epithelial ovarian cancer progression 75:111 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 4 32 theme desorption/ionization 844:864 arg1 spectrometry 886:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 1 33 from proteins 160:167 arg1 changes 123:129 Aberrant changes 114:129 Aberrant changes of N-glycan modifications on proteins 114:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 7 34 theme neutral 1716:1722 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 35 theme mass 881:884 arg1 spectrometry 886:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 1 36 from modifications 143:155 arg1 proteins 160:167 proteins 160:167 proteins 160:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 5 37 theme high-mannose 1105:1116 arg1 type 1118:1121 high-mannose type 1105:1121 high-mannose type 1105:1121 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 0 38 theme Mass 0:3 arg1 profiling 19:27 Mass spectrometric profiling 0:27 Mass spectrometric profiling 0:27 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 1 39 theme N-glycomic 302:311 arg1 analysis 313:320 N-glycomic analysis 302:320 N-glycomic analysis 302:320 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 4 40 from slides 809:814 arg1 N-glycans 754:762 N-glycans 754:762 N-glycans 754:762 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 5 41 theme N-glycan 1065:1072 arg1 compositions 1049:1060 the 80 compositions 1042:1060 the 80 compositions of N-glycan detected 1042:1081 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 4 42 theme matrix-assisted 822:836 arg1 spectrometry 886:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 4 43 theme different 933:941 arg1 grades 956:961 different pathological grades 933:961 different pathological grades of epithelial ovarian cancer 933:990 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 5 44 theme ovarian 1149:1155 arg1 samples 1164:1170 epithelial ovarian cancer samples 1138:1170 epithelial ovarian cancer samples 1138:1170 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 0 45 theme N-glycan 52:59 arg1 patterns 61:68 N-glycan patterns 52:68 N-glycan patterns 52:68 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 7 46 theme sialylated 1666:1675 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 3 47 from changes 489:495 arg1 patterns 500:507 patterns 500:507 patterns of N-glycans in tumor tissues 500:537 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 2 48 theme ovarian 370:376 arg1 development 385:395 epithelial ovarian cancer development 359:395 epithelial ovarian cancer development 359:395 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 28681696 1 49 from changes 123:129 arg1 proteins 160:167 proteins 160:167 proteins 160:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 1 50 theme possible 246:253 arg1 avenue 255:260 possible avenue 246:260 possible avenue for exploring their etiologies based on N-glycomic analysis 246:320 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 7 51 from changes 1762:1768 arg1 abundance 1782:1790 N-glycan abundance 1773:1790 N-glycan abundance 1773:1790 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 52 theme fucosylated 1650:1660 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 53 theme N-glycan 651:658 arg1 patterns 660:667 N-glycan patterns 651:667 N-glycan patterns 651:667 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 0 54 with association 37:47 arg1 progression 101:111 epithelial ovarian cancer progression 75:111 epithelial ovarian cancer progression 75:111 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 3 55 theme cancer 565:570 arg1 progression 572:582 epithelial ovarian cancer progression 546:582 epithelial ovarian cancer progression 546:582 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 8 56 theme strong 1863:1868 arg1 evidence 1870:1877 strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring 1863:2085 strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring 1863:2085 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 7 57 theme assay 1567:1571 arg1 analysis 1546:1553 a statistical analysis 1532:1553 a statistical analysis of variance assay 1532:1571 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 58 theme different 1588:1596 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 58 theme different 1588:1596 arg1 patterns 1607:1614 different N-glycan patterns 1588:1614 different N-glycan patterns 1588:1614 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 58 theme different 1588:1596 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 6 59 theme neutral 1404:1410 arg1 N-glycans 1420:1428 four high-mannose and three fucosylated neutral complex N-glycans 1364:1428 N-glycans 1420:1428 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 3 60 theme epithelial 546:555 arg1 cancer 565:570 epithelial ovarian cancer 546:570 epithelial ovarian cancer progression 546:582 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 7 61 theme N-glycan 1598:1605 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 61 theme N-glycan 1598:1605 arg1 patterns 1607:1614 different N-glycan patterns 1588:1614 different N-glycan patterns 1588:1614 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 61 theme N-glycan 1598:1605 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 62 from changes 640:646 arg1 patterns 660:667 N-glycan patterns 651:667 N-glycan patterns 651:667 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 3 63 from patterns 500:507 arg1 tissues 531:537 tumor tissues 525:537 tumor tissues 525:537 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 7 64 theme cancer 1818:1823 arg1 grades 1825:1830 epithelial ovarian cancer grades 1799:1830 epithelial ovarian cancer grades 1799:1830 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 65 theme ovarian 977:983 arg1 cancer 985:990 epithelial ovarian cancer 966:990 epithelial ovarian cancer 966:990 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 4 66 theme epithelial 715:724 arg1 cancer 734:739 epithelial ovarian cancer 715:739 epithelial ovarian cancer 715:739 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 6 67 theme healthy 1495:1501 arg1 controls 1503:1510 healthy controls 1495:1510 healthy controls 1495:1510 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 2 68 theme N-glycan 334:341 arg1 patterns 343:350 N-glycan patterns 334:350 N-glycan patterns 334:350 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 28681696 1 69 theme different 216:224 arg1 cancers 226:232 different cancers 216:232 different cancers 216:232 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 7 70 theme epithelial 1799:1808 arg1 grades 1825:1830 epithelial ovarian cancer grades 1799:1830 epithelial ovarian cancer grades 1799:1830 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 71 theme variance 1558:1565 arg1 assay 1567:1571 variance assay 1558:1571 variance assay 1558:1571 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 2 72 theme cancer 378:383 arg1 development 385:395 epithelial ovarian cancer development 359:395 epithelial ovarian cancer development 359:395 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 28681696 6 73 theme cancer 1483:1488 arg1 discrimination 1446:1459 good discrimination 1441:1459 good discrimination of epithelial ovarian cancer from healthy controls 1441:1510 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 6 74 theme epithelial 1464:1473 arg1 cancer 1483:1488 epithelial ovarian cancer 1464:1488 epithelial ovarian cancer 1464:1488 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 8 75 theme epithelial 1930:1939 arg1 grades 1969:1974 epithelial ovarian cancer pathological grades 1930:1974 epithelial ovarian cancer pathological grades 1930:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 6 76 gly fucosylated 1392:1402 arg1 N-glycans 1420:1428 four high-mannose and three fucosylated neutral complex N-glycans 1364:1428 N-glycans 1420:1428 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 8 77 theme cancer 1949:1954 arg1 grades 1969:1974 epithelial ovarian cancer pathological grades 1930:1974 epithelial ovarian cancer pathological grades 1930:1974 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 1 78 theme modifications 143:155 arg1 changes 123:129 Aberrant changes 114:129 Aberrant changes of N-glycan modifications on proteins 114:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 7 79 gly sialylated 1666:1675 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 7 80 theme specific 1753:1760 arg1 changes 1762:1768 specific changes 1753:1768 specific changes in N-glycan abundance across epithelial ovarian cancer grades 1753:1830 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 6 81 theme combined 1337:1344 arg1 panel 1346:1350 a combined panel 1335:1350 a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans 1335:1428 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 4 82 theme formalin-fixed 769:782 arg1 slides 809:814 formalin-fixed paraffin-embedded tissue slides 769:814 formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 769:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 0 83 theme epithelial 75:84 arg1 cancer 94:99 epithelial ovarian cancer 75:99 epithelial ovarian cancer progression 75:111 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 7 84 gly fucosylated 1650:1660 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 85 theme cancer 734:739 arg1 oncogenesis 684:694 oncogenesis 684:694 oncogenesis 684:694 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 4 85 theme cancer 734:739 arg1 progression 700:710 progression 700:710 progression 700:710 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 0 86 theme cancer 94:99 arg1 progression 101:111 epithelial ovarian cancer progression 75:111 epithelial ovarian cancer progression 75:111 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 5 87 theme glycans 1257:1263 arg1 levels 1235:1240 reduced levels 1227:1240 reduced levels of hybrid-type glycans 1227:1263 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 7 88 theme complex 1724:1730 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 8 89 theme novel 2015:2019 arg1 biomarkers 2021:2030 novel biomarkers 2015:2030 novel biomarkers for epithelial ovarian cancer diagnosis and monitoring 2015:2085 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 5 90 theme expression 1084:1093 arg1 levels 1095:1100 expression levels 1084:1100 expression levels of high-mannose type 1084:1121 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 7 91 theme fucosylated 1704:1714 arg1 N-glycans 1732:1740 10 fucosylated neutral complex N-glycans 1701:1740 10 fucosylated neutral complex N-glycans 1701:1740 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 4 92 theme laser 838:842 arg1 spectrometry 886:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 5 93 theme reduced 1227:1233 arg1 levels 1235:1240 reduced levels 1227:1240 reduced levels of hybrid-type glycans 1227:1263 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 4 94 theme time-of-flight 866:879 arg1 spectrometry 886:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 822:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 0 95 theme spectrometric 5:17 arg1 profiling 19:27 Mass spectrometric profiling 0:27 Mass spectrometric profiling 0:27 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 6 96 from controls 1503:1510 arg1 discrimination 1446:1459 good discrimination 1441:1459 good discrimination of epithelial ovarian cancer from healthy controls 1441:1510 By applying receiver operating characteristic analysis, we show that a combined panel composed of four high-mannose and three fucosylated neutral complex N-glycans allows for good discrimination of epithelial ovarian cancer from healthy controls. 28681696 2 97 theme cell 468:471 arg1 lines 473:477 cell lines 468:477 cell lines 468:477 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 28681696 4 98 theme tissue 802:807 arg1 slides 809:814 formalin-fixed paraffin-embedded tissue slides 769:814 formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 769:897 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 8 99 theme epithelial 2036:2045 arg1 diagnosis 2062:2070 epithelial ovarian cancer diagnosis 2036:2070 epithelial ovarian cancer diagnosis 2036:2070 Together, our results provide strong evidence that N-glycomic changes are a strong indicator for epithelial ovarian cancer pathological grades and should provide avenues to identify novel biomarkers for epithelial ovarian cancer diagnosis and monitoring. 28681696 1 100 theme Aberrant 114:121 arg1 changes 123:129 Aberrant changes 114:129 Aberrant changes of N-glycan modifications on proteins 114:167 Aberrant changes of N-glycan modifications on proteins have been linked to various diseases including different cancers, suggesting possible avenue for exploring their etiologies based on N-glycomic analysis. 28681696 5 101 theme epithelial 1138:1147 arg1 samples 1164:1170 epithelial ovarian cancer samples 1138:1170 epithelial ovarian cancer samples 1138:1170 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 0 102 theme patterns 61:68 arg1 association 37:47 association 37:47 association of N-glycan patterns with epithelial ovarian cancer progression 37:111 Mass spectrometric profiling reveals association of N-glycan patterns with epithelial ovarian cancer progression. 28681696 5 103 theme cancer 1157:1162 arg1 samples 1164:1170 epithelial ovarian cancer samples 1138:1170 epithelial ovarian cancer samples 1138:1170 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 4 104 theme pathological 943:954 arg1 grades 956:961 different pathological grades 933:961 different pathological grades of epithelial ovarian cancer 933:990 To investigate whether changes in N-glycan patterns correlate with oncogenesis and progression of epithelial ovarian cancer, we profiled N-glycans from formalin-fixed paraffin-embedded tissue slides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and quantitatively compared among different pathological grades of epithelial ovarian cancer and healthy controls. 28681696 3 105 theme N-glycans 512:520 arg1 patterns 500:507 patterns 500:507 patterns of N-glycans in tumor tissues 500:537 However, changes in patterns of N-glycans in tumor tissues during epithelial ovarian cancer progression have remained largely undefined. 28681696 7 106 theme complex 1677:1683 arg1 structures 1685:1694 2 fucosylated and sialylated complex structures 1648:1694 2 fucosylated and sialylated complex structures 1648:1694 Furthermore, using a statistical analysis of variance assay, we found that different N-glycan patterns, including 2 high-mannose-type, 2 fucosylated and sialylated complex structures, and 10 fucosylated neutral complex N-glycans, exhibited specific changes in N-glycan abundance across epithelial ovarian cancer grades. 28681696 5 107 theme type 1118:1121 arg1 levels 1095:1100 expression levels 1084:1100 expression levels of high-mannose type 1084:1121 Our results show that among the 80 compositions of N-glycan detected, expression levels of high-mannose type were higher in epithelial ovarian cancer samples than that observed in healthy controls, accompanied by reduced levels of hybrid-type glycans. 28681696 2 108 theme epithelial 359:368 arg1 development 385:395 epithelial ovarian cancer development 359:395 epithelial ovarian cancer development 359:395 Changes in N-glycan patterns during epithelial ovarian cancer development have so far been investigated mainly using serum, plasma, ascites, and cell lines. 27035801 2 0 theme anchimeric 417:426 arg1 assistance 428:437 anchimeric assistance 417:437 anchimeric assistance from a remote participating group 417:471 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 1 1 attach isolated 166:173 arg1 smegmatis 194:202 Mycobacterium smegmatis 180:202 Mycobacterium smegmatis 180:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 1 attach isolated 166:173 arg2 tetrasaccharides 149:164 three important trehalose containing tetrasaccharides 112:164 three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 112:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 3 2 from assembly 568:575 arg1 tuberculosis 626:637 M. tuberculosis 623:637 M. tuberculosis 623:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 1 3 theme stereoselective 292:306 arg1 glycosylations 308:321 stereoselective glycosylations 292:321 stereoselective glycosylations 292:321 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 3 4 theme M. 623:624 arg1 tuberculosis 626:637 M. tuberculosis 623:637 M. tuberculosis 623:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 1 5 theme important 118:126 arg1 tetrasaccharides 149:164 three important trehalose containing tetrasaccharides 112:164 three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 112:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 6 theme glycosylations 308:321 arg1 opening 257:263 regioselective opening 242:263 regioselective opening of benzylidene acetals and stereoselective glycosylations 242:321 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 6 theme glycosylations 308:321 arg1 steps 330:334 key steps 326:334 key steps 326:334 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 3 7 theme oligosaccharides 601:616 arg1 assembly 568:575 the assembly 564:575 the assembly of structurally related oligosaccharides from M. tuberculosis 564:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 1 8 theme Total 93:97 arg1 synthesis 99:107 Total synthesis 93:107 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 93:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 2 9 theme participating 453:465 arg1 group 467:471 a remote participating group 444:471 a remote participating group 444:471 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 0 10 theme total 6:10 arg1 synthesis 12:20 First total synthesis 0:20 First total synthesis of trehalose 0:33 First total synthesis of trehalose containing tetrasaccharides from Mycobacterium smegmatis. 27035801 3 11 theme related 593:599 arg1 oligosaccharides 601:616 structurally related oligosaccharides 580:616 structurally related oligosaccharides from M. tuberculosis 580:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 2 12 theme remote 446:451 arg1 group 467:471 a remote participating group 444:471 a remote participating group 444:471 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 0 13 theme First 0:4 arg1 synthesis 12:20 First total synthesis 0:20 First total synthesis of trehalose 0:33 First total synthesis of trehalose containing tetrasaccharides from Mycobacterium smegmatis. 27035801 1 14 theme key 326:328 arg1 opening 257:263 regioselective opening 242:263 regioselective opening of benzylidene acetals and stereoselective glycosylations 242:321 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 14 theme key 326:328 arg1 steps 330:334 key steps 326:334 key steps 326:334 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 15 theme first 224:228 arg1 time 230:233 the first time 220:233 the first time 220:233 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 2 16 theme solvent 493:499 arg1 participation 501:513 solvent participation 493:513 solvent participation 493:513 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 2 17 theme 1,2-cis 341:347 arg1 stereoselectivity 349:365 The 1,2-cis stereoselectivity 337:365 The 1,2-cis stereoselectivity in the glycosylation reactions 337:396 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 3 18 theme synthetic 520:528 arg1 strategy 530:537 The synthetic strategy 516:537 The synthetic strategy 516:537 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 3 19 used utilized 551:558 arg2 strategy 530:537 The synthetic strategy 516:537 The synthetic strategy 516:537 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 2 20 theme steric 474:479 arg1 effects 481:487 steric effects 474:487 steric effects 474:487 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 0 21 theme trehalose 25:33 arg1 synthesis 12:20 First total synthesis 0:20 First total synthesis of trehalose 0:33 First total synthesis of trehalose containing tetrasaccharides from Mycobacterium smegmatis. 27035801 1 22 theme regioselective 242:255 arg1 opening 257:263 regioselective opening 242:263 regioselective opening of benzylidene acetals and stereoselective glycosylations 242:321 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 22 theme regioselective 242:255 arg1 steps 330:334 key steps 326:334 key steps 326:334 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 23 theme trehalose 128:136 arg1 tetrasaccharides 149:164 three important trehalose containing tetrasaccharides 112:164 three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 112:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 2 24 from group 467:471 arg1 effects 481:487 steric effects 474:487 steric effects 474:487 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 2 24 from group 467:471 arg1 assistance 428:437 anchimeric assistance 417:437 anchimeric assistance from a remote participating group 417:471 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 2 24 from group 467:471 arg1 participation 501:513 solvent participation 493:513 solvent participation 493:513 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 1 25 theme containing 138:147 arg1 tetrasaccharides 149:164 three important trehalose containing tetrasaccharides 112:164 three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 112:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 26 theme tetrasaccharides 149:164 arg1 synthesis 99:107 Total synthesis 93:107 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis 93:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 2 27 theme glycosylation 374:386 arg1 reactions 388:396 the glycosylation reactions 370:396 the glycosylation reactions 370:396 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 3 28 from tuberculosis 626:637 arg1 oligosaccharides 601:616 structurally related oligosaccharides 580:616 structurally related oligosaccharides from M. tuberculosis 580:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 3 28 from tuberculosis 626:637 arg1 assembly 568:575 the assembly 564:575 the assembly of structurally related oligosaccharides from M. tuberculosis 564:637 The synthetic strategy can also be utilized for the assembly of structurally related oligosaccharides from M. tuberculosis. 27035801 1 29 theme benzylidene 268:278 arg1 acetals 280:286 benzylidene acetals 268:286 benzylidene acetals 268:286 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 0 30 theme Mycobacterium 68:80 arg1 smegmatis 82:90 Mycobacterium smegmatis 68:90 Mycobacterium smegmatis 68:90 First total synthesis of trehalose containing tetrasaccharides from Mycobacterium smegmatis. 27035801 1 31 theme Mycobacterium 180:192 arg1 smegmatis 194:202 Mycobacterium smegmatis 180:202 Mycobacterium smegmatis 180:202 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 2 32 from stereoselectivity 349:365 arg1 reactions 388:396 the glycosylation reactions 370:396 the glycosylation reactions 370:396 The 1,2-cis stereoselectivity in the glycosylation reactions was achieved using anchimeric assistance from a remote participating group, steric effects and solvent participation. 27035801 1 33 theme acetals 280:286 arg1 opening 257:263 regioselective opening 242:263 regioselective opening of benzylidene acetals and stereoselective glycosylations 242:321 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 27035801 1 33 theme acetals 280:286 arg1 steps 330:334 key steps 326:334 key steps 326:334 Total synthesis of three important trehalose containing tetrasaccharides isolated from Mycobacterium smegmatis is reported for the first time, using regioselective opening of benzylidene acetals and stereoselective glycosylations as key steps. 24912590 3 0 attach released 467:474 arg2 glycans 459:465 glycans 459:465 glycans released from proteins 459:488 The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. 24912590 3 0 attach released 467:474 arg1 proteins 481:488 proteins 481:488 proteins 481:488 The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. 24912590 1 1 theme plasma 143:148 arg1 samples 163:169 plasma and vitreous samples 143:169 samples 163:169 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 10 2 from profile 1424:1430 arg1 distinct 1464:1471 distinct 1464:1471 distinct 1464:1471 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 10 2 from profile 1424:1430 arg1 fluid 1448:1452 the vitreous fluid 1435:1452 the vitreous fluid 1435:1452 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 10 3 theme vitreous 1439:1446 arg1 distinct 1464:1471 distinct 1464:1471 distinct 1464:1471 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 10 3 theme vitreous 1439:1446 arg1 fluid 1448:1452 the vitreous fluid 1435:1452 the vitreous fluid 1435:1452 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 0 4 from comparison 2:11 arg1 fluid 63:67 vitreous fluid 54:67 vitreous fluid 54:67 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 0 4 from comparison 2:11 arg1 plasma 43:48 human plasma 37:48 human plasma 37:48 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 10 5 from that 1478:1481 arg1 distinct 1464:1471 distinct 1464:1471 distinct 1464:1471 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 10 5 from that 1478:1481 arg1 fluid 1448:1452 the vitreous fluid 1435:1452 the vitreous fluid 1435:1452 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 5 6 theme aminooxy-functionalized 598:620 arg1 aoWR 639:642 aoWR 639:642 aoWR 639:642 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 6 theme aminooxy-functionalized 598:620 arg1 reagent 630:636 aminooxy-functionalized peptide reagent 598:636 aminooxy-functionalized peptide reagent (aoWR) 598:643 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 7 attach released 649:656 arg2 glycans 573:579 Processed glycans 563:579 Processed glycans 563:579 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 7 attach released 649:656 arg1 beads 667:671 the beads 663:671 the beads 663:671 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 8 theme mass 758:761 arg1 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 8 theme mass 758:761 arg1 spectrometry 763:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 699:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 699:789 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 10 9 theme N-glycan 1415:1422 arg1 profile 1424:1430 N-glycan profile 1415:1430 N-glycan profile in the vitreous fluid, which is distinct from that in the plasma 1415:1495 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 7 10 theme 714 ± 29 pmol/100 μg 1018:1037 arg1 p < 0.001 1048:1056 p < 0.001 1048:1056 p < 0.001 1048:1056 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 7 10 theme 714 ± 29 pmol/100 μg 1018:1037 arg1 protein 1039:1045 714 ± 29 pmol/100 μg protein 1018:1045 714 ± 29 pmol/100 μg protein 1018:1045 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 3 11 theme BlotGlyco 507:515 arg1 beads 519:523 BlotGlyco H beads 507:523 BlotGlyco H beads 507:523 The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. 24912590 8 12 dep presumed 1184:1191 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 7 13 theme plasma 1002:1007 arg1 samples 1009:1015 plasma samples 1002:1015 plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001) 1002:1057 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 9 14 from N-glycan 1271:1278 arg1 samples 1292:1298 vitreous samples 1283:1298 vitreous samples (15.6 ± 1.5 %) 1283:1313 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 14 from N-glycan 1271:1278 arg1 % 1312:1312 15.6 ± 1.5 % 1301:1312 15.6 ± 1.5 % 1301:1312 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 1 15 dep concentration 97:109 arg1 the 93:95 the 93:95 the 93:95 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 7 16 theme vitreous 907:914 arg1 samples 916:922 vitreous samples 907:922 vitreous samples (132 ± 29 pmol/100 μg protein) 907:953 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 7 16 theme vitreous 907:914 arg1 protein 946:952 132 ± 29 pmol/100 μg protein 925:952 132 ± 29 pmol/100 μg protein 925:952 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 1 17 theme vitreous 154:161 arg1 samples 163:169 plasma and vitreous samples 143:169 samples 163:169 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 10 18 theme current 1372:1378 arg1 data 1380:1383 The current data 1368:1383 The current data 1368:1383 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 1 19 from composition 115:125 arg1 samples 163:169 plasma and vitreous samples 143:169 samples 163:169 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 8 20 theme Hex 1198:1200 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 21 theme NeuAc 1213:1217 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 22 theme Predominant 1060:1070 arg1 N-glycan 1072:1079 Predominant N-glycan 1060:1079 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) 1060:1143 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 23 theme vitreous 1115:1122 arg1 fluid 1124:1128 vitreous fluid 1115:1128 vitreous fluid (37.2 ± 3.1 %) 1115:1143 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 23 theme vitreous 1115:1122 arg1 % 1142:1142 37.2 ± 3.1 % 1131:1142 37.2 ± 3.1 % 1131:1142 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 7 24 theme N-glycans 894:902 arg1 Concentration 877:889 Concentration 877:889 Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) 877:953 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 5 25 theme desorption/ionization 721:741 arg1 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 25 theme desorption/ionization 721:741 arg1 spectrometry 763:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 699:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 699:789 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 1 26 from concentration 97:109 arg1 samples 163:169 plasma and vitreous samples 143:169 samples 163:169 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 2 27 theme macular 325:331 arg1 MH 339:340 MH 339:340 MH 339:340 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 27 theme macular 325:331 arg1 hole 333:336 idiopathic macular hole 314:336 idiopathic macular hole (MH) 314:341 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 8 28 theme 2+ 1219:1220 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 0 29 theme N-glycan 16:23 arg1 profiles 25:32 N-glycan profiles 16:32 N-glycan profiles 16:32 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 9 30 theme second-ranked 1257:1269 arg1 N-glycan 1271:1278 the second-ranked N-glycan 1253:1278 the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) 1253:1313 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 30 theme second-ranked 1257:1269 arg1 % 1352:1352 2.3 ± 0.2 % 1342:1352 2.3 ± 0.2 % 1342:1352 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 30 theme second-ranked 1257:1269 arg1 seventh 1323:1329 seventh 1323:1329 seventh 1323:1329 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 5 31 theme matrix-assisted 699:713 arg1 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 31 theme matrix-assisted 699:713 arg1 spectrometry 763:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 699:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 699:789 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 2 32 theme idiopathic 314:323 arg1 MH 339:340 MH 339:340 MH 339:340 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 32 theme idiopathic 314:323 arg1 hole 333:336 idiopathic macular hole 314:336 idiopathic macular hole (MH) 314:341 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 10 33 theme useful 1393:1398 arg1 information 1400:1410 useful information 1393:1410 useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma 1393:1495 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 6 34 from composition 814:824 arg1 samples 846:852 the samples 842:852 the samples 842:852 The concentration and composition of N-glycans in the samples were assessed. 24912590 5 35 theme laser 715:719 arg1 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 35 theme laser 715:719 arg1 spectrometry 763:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 699:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 699:789 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 9 36 from seventh 1323:1329 arg1 plasma 1334:1339 plasma 1334:1339 plasma 1334:1339 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 1 37 theme N-glycans 130:138 arg1 composition 115:125 composition 115:125 composition 115:125 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 1 37 theme N-glycans 130:138 arg1 concentration 97:109 concentration 97:109 concentration 97:109 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 1 38 with patients 185:192 arg1 diseases 231:238 non-proliferative vitreoretinal diseases 199:238 non-proliferative vitreoretinal diseases 199:238 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 6 39 dep concentration 796:808 arg1 The 792:794 The 792:794 The 792:794 The concentration and composition of N-glycans in the samples were assessed. 24912590 1 40 theme non-proliferative 199:215 arg1 diseases 231:238 non-proliferative vitreoretinal diseases 199:238 non-proliferative vitreoretinal diseases 199:238 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 0 41 theme profiles 25:32 arg1 comparison 2:11 A comparison 0:11 A comparison of N-glycan profiles in human plasma and vitreous fluid. 0:68 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 9 42 theme vitreous 1283:1290 arg1 samples 1292:1298 vitreous samples 1283:1298 vitreous samples (15.6 ± 1.5 %) 1283:1313 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 42 theme vitreous 1283:1290 arg1 % 1312:1312 15.6 ± 1.5 % 1301:1312 15.6 ± 1.5 % 1301:1312 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 5 43 theme time-of-flight 743:756 arg1 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 MALDI-TOF-MS 777:788 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 43 theme time-of-flight 743:756 arg1 spectrometry 763:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry 699:774 matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 699:789 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 1 44 theme vitreoretinal 217:229 arg1 diseases 231:238 non-proliferative vitreoretinal diseases 199:238 non-proliferative vitreoretinal diseases 199:238 PURPOSE To investigate the concentration and composition of N-glycans in plasma and vitreous samples obtained from patients with non-proliferative vitreoretinal diseases. 24912590 8 45 theme HexNAc 1204:1209 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 3 46 theme enzymatic 422:430 arg1 cleaving 432:439 enzymatic cleaving 422:439 enzymatic cleaving 422:439 The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. 24912590 0 47 theme human 37:41 arg1 plasma 43:48 human plasma 37:48 human plasma 37:48 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 9 48 from plasma 1334:1339 arg1 N-glycan 1271:1278 the second-ranked N-glycan 1253:1278 the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) 1253:1313 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 48 from plasma 1334:1339 arg1 % 1352:1352 2.3 ± 0.2 % 1342:1352 2.3 ± 0.2 % 1342:1352 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 48 from plasma 1334:1339 arg1 seventh 1323:1329 seventh 1323:1329 seventh 1323:1329 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 2 49 with patients 349:356 arg1 ERM 384:386 ERM 384:386 ERM 384:386 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 49 with patients 349:356 arg1 membrane 374:381 epiretinal membrane 363:381 epiretinal membrane (ERM) 363:387 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 49 with patients 349:356 arg1 MH 339:340 MH 339:340 MH 339:340 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 49 with patients 349:356 arg1 hole 333:336 idiopathic macular hole 314:336 idiopathic macular hole (MH) 314:341 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 7 50 from Concentration 877:889 arg1 samples 916:922 vitreous samples 907:922 vitreous samples (132 ± 29 pmol/100 μg protein) 907:953 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 7 50 from Concentration 877:889 arg1 protein 946:952 132 ± 29 pmol/100 μg protein 925:952 132 ± 29 pmol/100 μg protein 925:952 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 8 51 theme GlcNAc 1229:1234 arg1 Man 1223:1225 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3 1197:1227 (Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2 1197:1236 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 4 52 theme Sialic 526:531 arg1 acids 533:537 Sialic acids 526:537 Sialic acids 526:537 Sialic acids were methyl-esterified. 24912590 6 53 theme N-glycans 829:837 arg1 composition 814:824 composition 814:824 composition 814:824 The concentration and composition of N-glycans in the samples were assessed. 24912590 6 53 theme N-glycans 829:837 arg1 concentration 796:808 concentration 796:808 concentration 796:808 The concentration and composition of N-glycans in the samples were assessed. 24912590 5 54 theme peptide 622:628 arg1 aoWR 639:642 aoWR 639:642 aoWR 639:642 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 5 54 theme peptide 622:628 arg1 reagent 630:636 aminooxy-functionalized peptide reagent 598:636 aminooxy-functionalized peptide reagent (aoWR) 598:643 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 6 55 from concentration 796:808 arg1 samples 846:852 the samples 842:852 the samples 842:852 The concentration and composition of N-glycans in the samples were assessed. 24912590 7 56 dep samples 1009:1015 arg1 p < 0.001 1048:1056 p < 0.001 1048:1056 p < 0.001 1048:1056 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 7 56 dep samples 1009:1015 arg1 protein 1039:1045 714 ± 29 pmol/100 μg protein 1018:1045 714 ± 29 pmol/100 μg protein 1018:1045 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 5 57 theme Processed 563:571 arg1 glycans 573:579 Processed glycans 563:579 Processed glycans 563:579 Processed glycans were tagged with aminooxy-functionalized peptide reagent (aoWR) and released from the beads, followed by detection by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). 24912590 9 58 dep seventh 1323:1329 arg1 N-glycan 1271:1278 the second-ranked N-glycan 1253:1278 the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) 1253:1313 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 58 dep seventh 1323:1329 arg1 % 1352:1352 2.3 ± 0.2 % 1342:1352 2.3 ± 0.2 % 1342:1352 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 9 58 dep seventh 1323:1329 arg1 seventh 1323:1329 seventh 1323:1329 seventh 1323:1329 By contrast, the second-ranked N-glycan in vitreous samples (15.6 ± 1.5 %) was the seventh in plasma (2.3 ± 0.2 %). 24912590 8 59 from N-glycan 1072:1079 arg1 fluid 1124:1128 vitreous fluid 1115:1128 vitreous fluid (37.2 ± 3.1 %) 1115:1143 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 59 from N-glycan 1072:1079 arg1 % 1142:1142 37.2 ± 3.1 % 1131:1142 37.2 ± 3.1 % 1131:1142 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 59 from N-glycan 1072:1079 arg1 plasma 1089:1094 plasma 1089:1094 plasma (39.7 ± 1.1 %) 1089:1109 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 8 59 from N-glycan 1072:1079 arg1 % 1108:1108 39.7 ± 1.1 % 1097:1108 39.7 ± 1.1 % 1097:1108 Predominant N-glycan in both plasma (39.7 ± 1.1 %) and vitreous fluid (37.2 ± 3.1 %) was identical, and the composition was presumed as [(Hex)2(HexNAc)2(NeuAc)2+ (Man)3(GlcNAc)2]. 24912590 0 60 theme vitreous 54:61 arg1 fluid 63:67 vitreous fluid 54:67 vitreous fluid 54:67 A comparison of N-glycan profiles in human plasma and vitreous fluid. 24912590 2 61 dep METHODS 241:247 arg1 Plasma 249:254 Plasma 249:254 Plasma 249:254 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 61 dep METHODS 241:247 arg1 samples 269:275 vitreous samples 260:275 vitreous samples 260:275 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 62 theme vitreous 260:267 arg1 samples 269:275 vitreous samples 260:275 vitreous samples 260:275 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 63 theme epiretinal 363:372 arg1 ERM 384:386 ERM 384:386 ERM 384:386 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 63 theme epiretinal 363:372 arg1 membrane 374:381 epiretinal membrane 363:381 epiretinal membrane (ERM) 363:387 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 64 with patients 300:307 arg1 ERM 384:386 ERM 384:386 ERM 384:386 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 64 with patients 300:307 arg1 membrane 374:381 epiretinal membrane 363:381 epiretinal membrane (ERM) 363:387 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 64 with patients 300:307 arg1 MH 339:340 MH 339:340 MH 339:340 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 2 64 with patients 300:307 arg1 hole 333:336 idiopathic macular hole 314:336 idiopathic macular hole (MH) 314:341 METHODS Plasma and vitreous samples were collected from 11 patients with idiopathic macular hole (MH) and 9 patients with epiretinal membrane (ERM). 24912590 10 65 dep CONCLUSIONS 1356:1366 arg1 provide 1385:1391 provide 1385:1391 provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma 1385:1495 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 24912590 7 66 dep RESULTS 869:875 arg1 lower 973:977 lower 973:977 lower 973:977 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 3 67 theme H 517:517 arg1 beads 519:523 BlotGlyco H beads 507:523 BlotGlyco H beads 507:523 The samples were pretreated for enzymatic cleaving, and subsequently glycans released from proteins were captured on BlotGlyco H beads. 24912590 7 68 theme 132 ± 29 pmol/100 μg 925:944 arg1 samples 916:922 vitreous samples 907:922 vitreous samples (132 ± 29 pmol/100 μg protein) 907:953 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 7 68 theme 132 ± 29 pmol/100 μg 925:944 arg1 protein 946:952 132 ± 29 pmol/100 μg protein 925:952 132 ± 29 pmol/100 μg protein 925:952 RESULTS Concentration of N-glycans in vitreous samples (132 ± 29 pmol/100 μg protein) was significantly lower compared with those in plasma samples (714 ± 29 pmol/100 μg protein, p < 0.001). 24912590 10 69 from information 1400:1410 arg1 profile 1424:1430 N-glycan profile 1415:1430 N-glycan profile in the vitreous fluid, which is distinct from that in the plasma 1415:1495 CONCLUSIONS The current data provide useful information on N-glycan profile in the vitreous fluid, which is distinct from that in the plasma. 26085185 6 0 theme N-glycosylation 1053:1067 arg1 phenotypes 1069:1078 CRC-specific N-glycosylation phenotypes 1040:1078 CRC-specific N-glycosylation phenotypes 1040:1078 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 7 1 theme P 1412:1412 arg1 < 1414:1414 P < 0.05 1412:1419 P < 0.05 1412:1419 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 2 2 theme chromatography-tandem 394:414 arg1 platform 451:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform 378:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy 378:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 10 3 theme high 1793:1796 arg1 β1,4-GlcNAcylation 1808:1825 high bisecting β1,4-GlcNAcylation 1793:1825 high bisecting β1,4-GlcNAcylation 1793:1825 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 7 4 theme particular 1262:1271 arg1 α2,6-sialylation 1273:1288 particular α2,6-sialylation 1262:1288 particular α2,6-sialylation 1262:1288 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 3 5 theme tissues 664:670 arg1 proteomes 597:605 isolated membrane proteomes 579:605 isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 579:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 9 6 theme novel 1619:1623 arg1 link 1625:1628 a novel link 1617:1628 a novel link between the EGFR status and the N-glycosylation 1617:1676 Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. 26085185 3 7 theme non-tumorigenic 642:656 arg1 tissues 664:670 paired tumorigenic and adjacent non-tumorigenic colon tissues 610:670 tissues 664:670 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 1 8 theme structure-function 191:208 arg1 relationships 210:222 the structure-function relationships 187:222 the structure-function relationships of protein glycosylation 187:247 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 7 9 theme relative 1330:1337 arg1 tumors 1323:1328 CRC tumors 1319:1328 CRC tumors relative to non-tumorigenic tissues 1319:1364 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 10 10 theme low 1831:1833 arg1 α2,3-sialylation 1835:1850 low α2,3-sialylation 1831:1850 low α2,3-sialylation (both P < 0.05) 1831:1866 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 10 10 theme low 1831:1833 arg1 < 1860:1860 P < 0.05 1858:1865 P < 0.05 1858:1865 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 2 11 theme label-free 341:350 arg1 glycomics 365:373 label-free quantitative glycomics 341:373 label-free quantitative glycomics 341:373 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 9 12 theme EGFR 1642:1645 arg1 status 1647:1652 the EGFR status 1638:1652 the EGFR status 1638:1652 Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. 26085185 6 13 theme high 1129:1132 arg1 N-glycans 1175:1183 high mannose, hybrid and paucimannosidic type N-glycans 1129:1183 N-glycans 1175:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 8 14 theme CRC 1423:1425 arg1 N-glycosylation 1442:1456 CRC stage-specific N-glycosylation 1423:1456 CRC stage-specific N-glycosylation 1423:1456 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 0 15 theme protein 127:133 arg1 N-glycosylation 135:149 EGFR-specific protein N-glycosylation 113:149 EGFR-specific protein N-glycosylation 113:149 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 1 16 theme glycosylation 235:247 arg1 relationships 210:222 the structure-function relationships 187:222 the structure-function relationships of protein glycosylation 187:247 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 10 17 theme P 1858:1858 arg1 α2,3-sialylation 1835:1850 low α2,3-sialylation 1831:1850 low α2,3-sialylation (both P < 0.05) 1831:1866 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 10 17 theme P 1858:1858 arg1 < 1860:1860 P < 0.05 1858:1865 P < 0.05 1858:1865 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 5 18 theme extensive 975:983 arg1 heterogeneity 996:1008 an extensive structural heterogeneity 972:1008 an extensive structural heterogeneity with a total of 91 N-glycans 972:1037 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 4 19 theme epidermal 807:815 arg1 EGFR 841:844 EGFR 841:844 EGFR 841:844 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 4 19 theme epidermal 807:815 arg1 receptor 831:838 their epidermal growth factor receptor 801:838 their epidermal growth factor receptor (EGFR) status 801:852 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 3 20 theme males 694:698 arg1 cohort 679:684 a cohort 677:684 a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 677:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 4 21 theme factor 824:829 arg1 EGFR 841:844 EGFR 841:844 EGFR 841:844 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 4 21 theme factor 824:829 arg1 receptor 831:838 their epidermal growth factor receptor 801:838 their epidermal growth factor receptor (EGFR) status 801:852 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 11 22 theme EGFR 1958:1961 arg1 status 1963:1968 EGFR status 1958:1968 EGFR status 1958:1968 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 1 23 from markers 275:281 arg1 research 310:317 colorectal cancer (CRC) research 286:317 colorectal cancer (CRC) research 286:317 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 9 24 theme hierarchical 1699:1710 arg1 clustering 1712:1721 hierarchical clustering 1699:1721 hierarchical clustering of the N-glycome profiles 1699:1747 Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. 26085185 8 25 theme high 1474:1477 arg1 α2,3-sialylation 1479:1494 high α2,3-sialylation 1474:1494 high α2,3-sialylation 1474:1494 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 0 26 theme non-tumorigenic 61:75 arg1 tissues 77:83 non-tumorigenic tissues 61:83 non-tumorigenic tissues 61:83 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 6 27 theme type 1170:1173 arg1 N-glycans 1175:1183 high mannose, hybrid and paucimannosidic type N-glycans 1129:1183 N-glycans 1175:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 8 28 theme bisecting 1504:1512 arg1 β1,4-GlcNAcylation 1514:1531 low bisecting β1,4-GlcNAcylation 1500:1531 low bisecting β1,4-GlcNAcylation 1500:1531 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 11 29 theme N-glycan 1984:1991 arg1 features 1993:2000 specific N-glycan features 1975:2000 specific N-glycan features 1975:2000 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 5 30 theme N-glycans 1029:1037 arg1 total 1017:1021 a total 1015:1021 a total of 91 N-glycans 1015:1037 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 8 31 gly α2,3-sialylation 1479:1494 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 8 32 theme mid-late 1564:1571 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 0 33 theme In-depth 0:7 arg1 profiling 19:27 In-depth N-glycome profiling 0:27 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues 0:83 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 11 34 theme first 1919:1923 arg1 This 1907:1910 This 1907:1910 This 1907:1910 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 11 34 theme first 1919:1923 arg1 study 1925:1929 the first study 1915:1929 the first study to correlate CRC stage and EGFR status with specific N-glycan features 1915:2000 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 2 35 theme CRC 526:528 arg1 malignancy 530:539 CRC malignancy 526:539 CRC malignancy 526:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 5 36 theme relative 939:946 arg1 quantitation 948:959 relative quantitation 939:959 relative quantitation 939:959 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 6 37 theme complex 1216:1222 arg1 N-glycans 1224:1232 complex N-glycans 1216:1232 complex N-glycans (P < 0.05) 1216:1243 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 6 37 theme complex 1216:1222 arg1 < 1237:1237 P < 0.05 1235:1242 P < 0.05 1235:1242 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 7 38 dep down-regulated 1396:1409 arg1 < 1414:1414 P < 0.05 1412:1419 P < 0.05 1412:1419 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 2 39 theme N-glycosylation 486:500 arg1 changes 502:508 the N-glycosylation changes 482:508 the N-glycosylation changes associated with CRC malignancy 482:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 3 40 theme N-Glycome 542:550 arg1 profiling 552:560 N-Glycome profiling 542:560 N-Glycome profiling 542:560 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 0 41 theme paired 32:37 arg1 cancer 50:55 paired colorectal cancer 32:55 paired colorectal cancer 32:55 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 3 42 theme membrane 588:595 arg1 proteomes 597:605 isolated membrane proteomes 579:605 isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 579:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 5 43 theme Detailed 900:907 arg1 characterization 918:933 Detailed N-glycan characterization 900:933 Detailed N-glycan characterization 900:933 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 11 44 theme CRC 1944:1946 arg1 stage 1948:1952 CRC stage 1944:1952 CRC stage 1944:1952 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 1 45 theme glycoprotein 262:273 arg1 markers 275:281 glycoprotein markers 262:281 glycoprotein markers in colorectal cancer (CRC) research 262:317 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 10 46 theme N-glycan 1764:1771 arg1 signatures 1773:1782 EGFR-specific N-glycan signatures 1750:1782 EGFR-specific N-glycan signatures 1750:1782 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 3 47 theme tumorigenic 617:627 arg1 proteomes 597:605 isolated membrane proteomes 579:605 isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 579:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 8 48 from β1,4-GlcNAcylation 1514:1531 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 2 49 theme mass 416:419 arg1 platform 451:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform 378:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy 378:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 1 50 theme cancer 297:302 arg1 research 310:317 colorectal cancer (CRC) research 286:317 colorectal cancer (CRC) research 286:317 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 6 51 theme CRC-specific 1040:1051 arg1 phenotypes 1069:1078 CRC-specific N-glycosylation phenotypes 1040:1078 CRC-specific N-glycosylation phenotypes 1040:1078 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 10 52 theme bisecting 1798:1806 arg1 β1,4-GlcNAcylation 1808:1825 high bisecting β1,4-GlcNAcylation 1793:1825 high bisecting β1,4-GlcNAcylation 1793:1825 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 10 53 dep < 1860:1860 arg1 both 1853:1856 both 1853:1856 both 1853:1856 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 3 54 theme colon 658:662 arg1 tissues 664:670 paired tumorigenic and adjacent non-tumorigenic colon tissues 610:670 tissues 664:670 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 2 55 theme carbon-liquid 380:392 arg1 platform 451:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform 378:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy 378:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 3 56 theme adjacent 633:640 arg1 tissues 664:670 paired tumorigenic and adjacent non-tumorigenic colon tissues 610:670 tissues 664:670 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 5 57 theme structural 985:994 arg1 heterogeneity 996:1008 an extensive structural heterogeneity 972:1008 an extensive structural heterogeneity with a total of 91 N-glycans 972:1037 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 0 58 theme cancer 50:55 arg1 profiling 19:27 In-depth N-glycome profiling 0:27 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues 0:83 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 10 59 theme CRC 1894:1896 arg1 tissues 1898:1904 EGFR-negative CRC tissues 1880:1904 EGFR-negative CRC tissues 1880:1904 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 8 60 theme Lewis-type 1537:1546 arg1 fucosylation 1548:1559 Lewis-type fucosylation 1537:1559 Lewis-type fucosylation 1537:1559 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 2 61 theme quantitative 352:363 arg1 glycomics 365:373 label-free quantitative glycomics 341:373 label-free quantitative glycomics 341:373 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 6 62 theme mannose 1134:1140 arg1 hybrid 1143:1148 high mannose, hybrid and paucimannosidic type N-glycans 1129:1183 hybrid 1143:1148 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 4 63 theme CRC 765:767 arg1 tissues 769:775 The CRC tissues 761:775 The CRC tissues 761:775 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 3 64 from proteomes 597:605 arg1 cohort 679:684 a cohort 677:684 a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 677:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 1 65 theme protein 227:233 arg1 glycosylation 235:247 protein glycosylation 227:247 protein glycosylation 227:247 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 9 66 theme N-glycome 1730:1738 arg1 profiles 1740:1747 the N-glycome profiles 1726:1747 the N-glycome profiles 1726:1747 Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. 26085185 11 67 theme biomolecular 2066:2077 arg1 deregulation 2079:2090 the biomolecular deregulation 2062:2090 the biomolecular deregulation associated with CRC 2062:2110 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 8 68 theme stage-specific 1427:1440 arg1 N-glycosylation 1442:1456 CRC stage-specific N-glycosylation 1423:1456 CRC stage-specific N-glycosylation 1423:1456 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 11 69 theme mechanisms 2043:2052 arg1 understanding 2022:2034 our understanding 2018:2034 our understanding of the mechanisms causing the biomolecular deregulation associated with CRC 2018:2110 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 4 70 theme growth 817:822 arg1 EGFR 841:844 EGFR 841:844 EGFR 841:844 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 4 70 theme growth 817:822 arg1 receptor 831:838 their epidermal growth factor receptor 801:838 their epidermal growth factor receptor (EGFR) status 801:852 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 3 71 from cohort 679:684 arg1 proteomes 597:605 isolated membrane proteomes 579:605 isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 579:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 3 71 from cohort 679:684 arg1 tumorigenic 617:627 tumorigenic 617:627 tumorigenic 617:627 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 0 72 theme tissues 77:83 arg1 profiling 19:27 In-depth N-glycome profiling 0:27 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues 0:83 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 3 73 theme ± 706:706 arg1 y.o. 713:716 62.6 ± 13.1 y.o. 701:716 62.6 ± 13.1 y.o. 701:716 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 3 73 theme ± 706:706 arg1 males 694:698 five males 689:698 five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 689:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 6 74 theme N-glycans 1175:1183 arg1 under-representation 1192:1211 an under-representation 1189:1211 an under-representation of complex N-glycans (P < 0.05) 1189:1243 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 6 74 theme N-glycans 1175:1183 arg1 overrepresentation 1107:1124 an overrepresentation 1104:1124 an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans 1104:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 6 75 theme paucimannosidic 1154:1168 arg1 N-glycans 1175:1183 high mannose, hybrid and paucimannosidic type N-glycans 1129:1183 N-glycans 1175:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 8 76 theme low 1500:1502 arg1 β1,4-GlcNAcylation 1514:1531 low bisecting β1,4-GlcNAcylation 1500:1531 low bisecting β1,4-GlcNAcylation 1500:1531 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 1 77 gly glycoprotein 262:273 arg1 glycoprotein 262:273 glycoprotein markers 262:281 glycoprotein markers in colorectal cancer (CRC) research 262:317 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 9 78 theme profiles 1740:1747 arg1 clustering 1712:1721 hierarchical clustering 1699:1721 hierarchical clustering of the N-glycome profiles 1699:1747 Interestingly, a novel link between the EGFR status and the N-glycosylation was identified using hierarchical clustering of the N-glycome profiles. 26085185 6 79 theme hybrid 1143:1148 arg1 N-glycans 1175:1183 high mannose, hybrid and paucimannosidic type N-glycans 1129:1183 N-glycans 1175:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 11 80 theme specific 1975:1982 arg1 features 1993:2000 specific N-glycan features 1975:2000 specific N-glycan features 1975:2000 This is the first study to correlate CRC stage and EGFR status with specific N-glycan features, thus advancing our understanding of the mechanisms causing the biomolecular deregulation associated with CRC. 26085185 0 81 theme EGFR-specific 113:125 arg1 N-glycosylation 135:149 EGFR-specific protein N-glycosylation 113:149 EGFR-specific protein N-glycosylation 113:149 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 4 82 theme western 857:863 arg1 blotting 865:872 western blotting 857:872 western blotting 857:872 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 7 83 theme non-tumorigenic 1342:1356 arg1 tissues 1358:1364 non-tumorigenic tissues 1342:1364 non-tumorigenic tissues 1342:1364 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 5 84 theme N-glycan 909:916 arg1 characterization 918:933 Detailed N-glycan characterization 900:933 Detailed N-glycan characterization 900:933 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 6 85 theme P 1235:1235 arg1 N-glycans 1224:1232 complex N-glycans 1216:1232 complex N-glycans (P < 0.05) 1216:1243 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 6 85 theme P 1235:1235 arg1 < 1237:1237 P < 0.05 1235:1242 P < 0.05 1235:1242 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 4 86 theme receptor 831:838 arg1 status 847:852 their epidermal growth factor receptor (EGFR) status 801:852 their epidermal growth factor receptor (EGFR) status 801:852 The CRC tissues were typed according to their epidermal growth factor receptor (EGFR) status by western blotting and immunohistochemistry. 26085185 7 87 theme CRC 1319:1321 arg1 tumors 1323:1328 CRC tumors 1319:1328 CRC tumors relative to non-tumorigenic tissues 1319:1364 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 8 88 from α2,3-sialylation 1479:1494 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 6 89 theme N-glycans 1224:1232 arg1 under-representation 1192:1211 an under-representation 1189:1211 an under-representation of complex N-glycans (P < 0.05) 1189:1243 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 6 89 theme N-glycans 1224:1232 arg1 overrepresentation 1107:1124 an overrepresentation 1104:1124 an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans 1104:1183 CRC-specific N-glycosylation phenotypes were observed including an overrepresentation of high mannose, hybrid and paucimannosidic type N-glycans and an under-representation of complex N-glycans (P < 0.05). 26085185 7 90 from Sialylation 1246:1256 arg1 α2,6-sialylation 1273:1288 particular α2,6-sialylation 1262:1288 particular α2,6-sialylation 1262:1288 Sialylation, in particular α2,6-sialylation, was significantly higher in CRC tumors relative to non-tumorigenic tissues, whereas α2,3-sialylation was down-regulated (P < 0.05). 26085185 3 91 theme colorectal 734:743 arg1 adenocarcinoma 745:758 colorectal adenocarcinoma 734:758 colorectal adenocarcinoma 734:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 8 92 theme relative 1573:1580 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 2 93 theme analytical 440:449 arg1 platform 451:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform 378:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy 378:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 0 94 theme colorectal 39:48 arg1 cancer 50:55 paired colorectal cancer 32:55 paired colorectal cancer 32:55 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 8 95 gly fucosylation 1548:1559 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 8 96 theme early 1585:1589 arg1 stage 1591:1595 early stage 1585:1595 early stage 1585:1595 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 10 97 theme EGFR-negative 1880:1892 arg1 tissues 1898:1904 EGFR-negative CRC tissues 1880:1904 EGFR-negative CRC tissues 1880:1904 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 2 98 theme spectrometry-based 421:438 arg1 platform 451:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform 378:458 a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy 378:539 Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography-tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. 26085185 8 99 from fucosylation 1548:1559 arg1 CRC 1597:1599 mid-late relative to early stage CRC 1564:1599 mid-late relative to early stage CRC 1564:1599 CRC stage-specific N-glycosylation was detected by high α2,3-sialylation and low bisecting β1,4-GlcNAcylation and Lewis-type fucosylation in mid-late relative to early stage CRC. 26085185 5 100 with heterogeneity 996:1008 arg1 total 1017:1021 a total 1015:1021 a total of 91 N-glycans 1015:1037 Detailed N-glycan characterization and relative quantitation identified an extensive structural heterogeneity with a total of 91 N-glycans. 26085185 3 101 theme isolated 579:586 arg1 proteomes 597:605 isolated membrane proteomes 579:605 isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma 579:758 N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. 26085185 10 102 theme relative 1868:1875 arg1 β1,4-GlcNAcylation 1808:1825 high bisecting β1,4-GlcNAcylation 1793:1825 high bisecting β1,4-GlcNAcylation 1793:1825 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 10 103 theme EGFR-specific 1750:1762 arg1 signatures 1773:1782 EGFR-specific N-glycan signatures 1750:1782 EGFR-specific N-glycan signatures 1750:1782 EGFR-specific N-glycan signatures included high bisecting β1,4-GlcNAcylation and low α2,3-sialylation (both P < 0.05) relative to EGFR-negative CRC tissues. 26085185 0 104 theme N-glycome 9:17 arg1 profiling 19:27 In-depth N-glycome profiling 0:27 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues 0:83 In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation. 26085185 1 105 theme colorectal 286:295 arg1 CRC 305:307 CRC 305:307 CRC 305:307 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 26085185 1 105 theme colorectal 286:295 arg1 cancer 297:302 colorectal cancer 286:302 colorectal cancer (CRC) research 286:317 Glycomics may assist in uncovering the structure-function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. 24685714 3 0 theme cell 632:635 arg1 walls 637:641 cell walls 632:641 cell walls of etiolated hypocotyls 632:665 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 1 1 theme Pro-rich 377:384 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 1 theme Pro-rich 377:384 arg1 domain 386:391 a Pro-rich domain 375:391 a Pro-rich domain 375:391 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 5 2 from function 889:896 arg1 walls 906:910 cell walls 901:910 cell walls 901:910 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 15 3 theme organs 2223:2228 arg1 walls 2198:2202 cell walls 2193:2202 cell walls of quickly growing organs such as etiolated hypocotyls 2193:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 14 4 theme complex 2102:2108 arg1 scaffolds 2126:2134 complex supra-molecular scaffolds 2102:2134 complex supra-molecular scaffolds 2102:2134 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 3 5 theme hypocotyls 656:665 arg1 walls 637:641 cell walls 632:641 cell walls of etiolated hypocotyls 632:665 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 1 6 theme PAC 399:401 arg1 domain 428:433 a PAC (PRP-AGP containing Cys) domain 397:433 a PAC (PRP-AGP containing Cys) domain 397:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 6 theme PAC 399:401 arg1 Cys 423:425 PRP-AGP containing Cys 404:425 PRP-AGP containing Cys 404:425 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 6 theme PAC 399:401 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 7 theme PRP-AGP 404:410 arg1 domain 428:433 a PAC (PRP-AGP containing Cys) domain 397:433 a PAC (PRP-AGP containing Cys) domain 397:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 7 theme PRP-AGP 404:410 arg1 Cys 423:425 PRP-AGP containing Cys 404:425 PRP-AGP containing Cys 404:425 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 5 8 theme present 815:821 arg1 study 823:827 the present study 811:827 the present study 811:827 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 14 9 dep CONCLUSIONS 1953:1963 arg1 allow 1979:1983 allow 1979:1983 allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds 1979:2134 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 6 10 theme Brachypodium 1096:1107 arg1 distachyon 1109:1118 Brachypodium distachyon 1096:1118 Brachypodium distachyon 1096:1118 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 10 11 theme first 1526:1530 arg1 evidence 1545:1552 the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates 1522:1659 the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates 1522:1659 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 10 11 theme first 1526:1530 arg1 This 1514:1517 This 1514:1517 This 1514:1517 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 11 12 theme polygalacturonic 1708:1723 arg1 acid 1725:1728 methylesterified polygalacturonic acid 1691:1728 methylesterified polygalacturonic acid 1691:1728 AGP31 was also found to bind methylesterified polygalacturonic acid, possibly through its His-stretch. 24685714 1 13 from thaliana 263:270 arg1 unique 241:246 unique 241:246 unique 241:246 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 14 theme plant 178:182 arg1 protein 143:149 Arabinogalactan protein 31 127:152 Arabinogalactan protein 31 (AGP31) 127:160 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 14 theme plant 178:182 arg1 protein 194:200 a remarkable plant cell-wall protein 165:200 a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana 165:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 0 15 from protein 63:69 arg1 walls 100:104 Arabidopsis thaliana cell walls 74:104 Arabidopsis thaliana cell walls 74:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 1 16 theme acids 353:357 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 16 theme acids 353:357 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 16 theme acids 353:357 arg1 domain 331:336 a short AGP domain 319:336 a short AGP domain of seven amino acids 319:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 14 17 theme cell-wall 2049:2057 arg1 components 2059:2068 different cell-wall components 2039:2068 different cell-wall components 2039:2068 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 2 18 theme AGP 515:517 arg1 domain 519:524 the AGP domain 511:524 the AGP domain 511:524 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 9 19 theme PAC 1445:1447 arg1 domain 1449:1454 its PAC domain 1441:1454 its PAC domain with galactans that are branches of rhamnogalacturonan I 1441:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 0 20 theme putative 38:45 arg1 protein 63:69 a putative network-forming protein 36:69 a putative network-forming protein in Arabidopsis thaliana cell walls 36:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 0 20 theme putative 38:45 arg1 protein 16:22 Arabinogalactan protein 31 0:25 Arabinogalactan protein 31 (AGP31) 0:33 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 14 21 theme different 2039:2047 arg1 components 2059:2068 different cell-wall components 2039:2068 different cell-wall components 2039:2068 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 6 22 theme Nitrocellulose 921:934 arg1 membranes 936:944 Nitrocellulose membranes 921:944 Nitrocellulose membranes 921:944 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 2 23 theme O-glycosylation 461:475 arg1 patterns 477:484 different O-glycosylation patterns 451:484 different O-glycosylation patterns 451:484 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 14 24 theme AGP31 2028:2032 arg1 interactions 2012:2023 interactions 2012:2023 interactions of AGP31 with different cell-wall components 2012:2068 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 8 25 theme light 1300:1304 arg1 scattering 1306:1315 dynamic light scattering 1292:1315 dynamic light scattering (DLS) analyses 1292:1330 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 8 25 theme light 1300:1304 arg1 DLS 1318:1320 DLS 1318:1320 DLS 1318:1320 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 0 26 theme Arabidopsis 74:84 arg1 walls 100:104 Arabidopsis thaliana cell walls 74:104 Arabidopsis thaliana cell walls 74:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 1 27 theme short 321:325 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 27 theme short 321:325 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 27 theme short 321:325 arg1 domain 331:336 a short AGP domain 319:336 a short AGP domain of seven amino acids 319:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 14 28 theme interactions 2012:2023 arg1 model 2003:2007 a model 2001:2007 a model 2001:2007 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 0 29 theme cell 95:98 arg1 walls 100:104 Arabidopsis thaliana cell walls 74:104 Arabidopsis thaliana cell walls 74:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 14 30 theme model 2003:2007 arg1 proposal 1989:1996 the proposal 1985:1996 the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds 1985:2134 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 0 31 theme Arabinogalactan 0:14 arg1 protein 63:69 a putative network-forming protein 36:69 a putative network-forming protein in Arabidopsis thaliana cell walls 36:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 0 31 theme Arabinogalactan 0:14 arg1 protein 16:22 Arabinogalactan protein 31 0:25 Arabinogalactan protein 31 (AGP31) 0:33 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 0 31 theme Arabinogalactan 0:14 arg1 AGP31 28:32 AGP31 28:32 AGP31 28:32 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 15 32 theme cell 2193:2196 arg1 walls 2198:2202 cell walls 2193:2202 cell walls of quickly growing organs such as etiolated hypocotyls 2193:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 1 33 theme Arabinogalactan 127:141 arg1 protein 143:149 Arabinogalactan protein 31 127:152 Arabinogalactan protein 31 (AGP31) 127:160 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 33 theme Arabinogalactan 127:141 arg1 AGP31 155:159 AGP31 155:159 AGP31 155:159 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 33 theme Arabinogalactan 127:141 arg1 protein 194:200 a remarkable plant cell-wall protein 165:200 a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana 165:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 domain 386:391 a Pro-rich domain 375:391 a Pro-rich domain 375:391 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 domain 331:336 a short AGP domain 319:336 a short AGP domain of seven amino acids 319:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 domain 428:433 a PAC (PRP-AGP containing Cys) domain 397:433 a PAC (PRP-AGP containing Cys) domain 397:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 His-stretch 362:372 a His-stretch 360:372 a His-stretch 360:372 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 34 theme predicted 288:296 arg1 SP 314:315 SP 314:315 SP 314:315 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 6 35 theme different 972:980 arg1 polysaccharides 982:996 spotting different polysaccharides 963:996 spotting different polysaccharides 963:996 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 10 36 theme homologues 1626:1635 arg1 protein 1597:1603 an entire protein 1587:1603 an entire protein 1587:1603 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 10 36 theme homologues 1626:1635 arg1 domain 1610:1615 a domain 1608:1615 a domain of AGP31 homologues 1608:1635 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 10 36 theme homologues 1626:1635 arg1 homologues 1626:1635 AGP31 homologues 1620:1635 AGP31 homologues 1620:1635 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 7 37 theme native 1237:1242 arg1 AGP31 1244:1248 native AGP31 1237:1248 native AGP31 1237:1248 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 7 38 dep in 1153:1154 arg1 vitro 1156:1160 vitro 1156:1160 vitro 1156:1160 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 13 39 theme DLS 1846:1848 arg1 data 1850:1853 DLS data 1846:1853 DLS data 1846:1853 DLS data showed that AGP31 forms aggregates in solution, corroborating the hypothesis of an auto-assembly. 24685714 8 40 theme purified 1361:1368 arg1 fraction 1370:1377 an AGP31 purified fraction 1352:1377 an AGP31 purified fraction 1352:1377 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 13 41 theme auto-assembly 1938:1950 arg1 hypothesis 1921:1930 the hypothesis 1917:1930 the hypothesis of an auto-assembly 1917:1950 DLS data showed that AGP31 forms aggregates in solution, corroborating the hypothesis of an auto-assembly. 24685714 3 42 theme abundant 612:619 arg1 protein 621:627 an abundant protein 609:627 an abundant protein in cell walls of etiolated hypocotyls 609:665 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 3 42 theme abundant 612:619 arg1 AGP31 580:584 AGP31 580:584 AGP31 580:584 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 1 43 theme multi-domain 215:226 arg1 organization 228:239 a multi-domain organization 213:239 a multi-domain organization unique in Arabidopsis thaliana 213:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 2 44 from arabinogalactans 491:506 arg1 domain 519:524 the AGP domain 511:524 the AGP domain 511:524 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 2 44 from arabinogalactans 491:506 arg1 motifs 549:554 Hyp-O-Gal/Ara-rich motifs 530:554 Hyp-O-Gal/Ara-rich motifs 530:554 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 3 45 from protein 621:627 arg1 walls 637:641 cell walls 632:641 cell walls of etiolated hypocotyls 632:665 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 3 46 theme etiolated 646:654 arg1 hypocotyls 656:665 etiolated hypocotyls 646:665 etiolated hypocotyls 646:665 AGP31 has been identified as an abundant protein in cell walls of etiolated hypocotyls, but its function has not been investigated thus far. 24685714 1 47 theme unique 241:246 arg1 organization 228:239 a multi-domain organization 213:239 a multi-domain organization unique in Arabidopsis thaliana 213:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 48 theme Arabidopsis 251:261 arg1 thaliana 263:270 Arabidopsis thaliana 251:270 Arabidopsis thaliana 251:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 2 49 theme Hyp-O-Gal/Ara-rich 530:547 arg1 motifs 549:554 Hyp-O-Gal/Ara-rich motifs 530:554 Hyp-O-Gal/Ara-rich motifs 530:554 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 2 50 from domain 519:524 arg1 domain 572:577 the Pro-rich domain 559:577 the Pro-rich domain 559:577 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 14 51 theme supra-molecular 2110:2124 arg1 scaffolds 2126:2134 complex supra-molecular scaffolds 2102:2134 complex supra-molecular scaffolds 2102:2134 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24685714 6 52 theme thaliana 1083:1090 arg1 walls 1062:1066 cell walls 1057:1066 cell walls of Arabidopsis thaliana and Brachypodium distachyon 1057:1118 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 15 53 theme Such 2137:2140 arg1 scaffolds 2142:2150 Such scaffolds 2137:2150 Such scaffolds 2137:2150 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 9 54 theme KEY 1380:1382 arg1 RESULTS 1384:1390 KEY RESULTS 1380:1390 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 1380:1512 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 4 55 theme cell-wall 774:782 arg1 components 784:793 cell-wall components 774:793 cell-wall components 774:793 Literature data suggest that AGP31 may interact with cell-wall components. 24685714 2 56 theme Pro-rich 563:570 arg1 domain 572:577 the Pro-rich domain 559:577 the Pro-rich domain 559:577 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 6 57 theme distachyon 1109:1118 arg1 walls 1062:1066 cell walls 1057:1066 cell walls of Arabidopsis thaliana and Brachypodium distachyon 1057:1118 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 10 58 theme experimental 1532:1543 arg1 evidence 1545:1552 the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates 1522:1659 the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates 1522:1659 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 10 58 theme experimental 1532:1543 arg1 This 1514:1517 This 1514:1517 This 1514:1517 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 1 59 theme containing 412:421 arg1 domain 428:433 a PAC (PRP-AGP containing Cys) domain 397:433 a PAC (PRP-AGP containing Cys) domain 397:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 59 theme containing 412:421 arg1 Cys 423:425 PRP-AGP containing Cys 404:425 PRP-AGP containing Cys 404:425 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 60 theme remarkable 167:176 arg1 protein 143:149 Arabinogalactan protein 31 127:152 Arabinogalactan protein 31 (AGP31) 127:160 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 60 theme remarkable 167:176 arg1 protein 194:200 a remarkable plant cell-wall protein 165:200 a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana 165:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 6 61 theme Arabidopsis 1071:1081 arg1 thaliana 1083:1090 Arabidopsis thaliana 1071:1090 Arabidopsis thaliana 1071:1090 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 1 62 from unique 241:246 arg1 thaliana 263:270 Arabidopsis thaliana 251:270 Arabidopsis thaliana 251:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 63 theme amino 347:351 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 64 theme cell-wall 184:192 arg1 protein 143:149 Arabinogalactan protein 31 127:152 Arabinogalactan protein 31 (AGP31) 127:160 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 64 theme cell-wall 184:192 arg1 protein 194:200 a remarkable plant cell-wall protein 165:200 a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana 165:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 10 65 theme PAC 1561:1563 arg1 domain 1565:1570 a PAC domain 1559:1570 a PAC domain 1559:1570 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 7 66 theme in 1153:1154 arg1 assays 1174:1179 in vitro interaction assays 1153:1179 in vitro interaction assays 1153:1179 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 5 67 theme AGP31 845:849 arg1 partners 851:858 AGP31 partners 845:858 AGP31 partners 845:858 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 8 68 dep purified 1361:1368 arg1 AGP31 1355:1359 AGP31 1355:1359 AGP31 1355:1359 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 6 69 theme spotting 963:970 arg1 polysaccharides 982:996 spotting different polysaccharides 963:996 spotting different polysaccharides 963:996 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 0 70 theme network-forming 47:61 arg1 protein 63:69 a putative network-forming protein 36:69 a putative network-forming protein in Arabidopsis thaliana cell walls 36:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 0 70 theme network-forming 47:61 arg1 protein 16:22 Arabinogalactan protein 31 0:25 Arabinogalactan protein 31 (AGP31) 0:33 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 5 71 theme study 823:827 arg1 purpose 800:806 The purpose 796:806 The purpose of the present study 796:827 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 6 72 dep METHODS 913:919 arg1 prepared 951:958 prepared 951:958 were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon 946:1118 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 7 73 theme interaction 1162:1172 arg1 assays 1174:1179 in vitro interaction assays 1153:1179 in vitro interaction assays 1153:1179 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 12 74 theme PAC 1834:1836 arg1 domain 1838:1843 its PAC domain 1830:1843 its PAC domain 1830:1843 Finally, AGP31 was able to interact with itself in vitro through its PAC domain. 24685714 11 75 theme methylesterified 1691:1706 arg1 acid 1725:1728 methylesterified polygalacturonic acid 1691:1728 methylesterified polygalacturonic acid 1691:1728 AGP31 was also found to bind methylesterified polygalacturonic acid, possibly through its His-stretch. 24685714 0 76 theme thaliana 86:93 arg1 walls 100:104 Arabidopsis thaliana cell walls 74:104 Arabidopsis thaliana cell walls 74:104 Arabinogalactan protein 31 (AGP31), a putative network-forming protein in Arabidopsis thaliana cell walls? 24685714 2 77 theme different 451:459 arg1 patterns 477:484 different O-glycosylation patterns 451:484 different O-glycosylation patterns 451:484 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 8 78 theme dynamic 1292:1298 arg1 scattering 1306:1315 dynamic light scattering 1292:1315 dynamic light scattering (DLS) analyses 1292:1330 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 8 78 theme dynamic 1292:1298 arg1 DLS 1318:1320 DLS 1318:1320 DLS 1318:1320 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 7 79 theme arrays 1145:1150 arg1 validation 1127:1136 validation 1127:1136 validation of the arrays 1127:1150 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 1 80 theme AGP 327:329 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 80 theme AGP 327:329 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 80 theme AGP 327:329 arg1 domain 331:336 a short AGP domain 319:336 a short AGP domain of seven amino acids 319:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 9 81 theme rhamnogalacturonan 1492:1509 arg1 galactans 1461:1469 galactans 1461:1469 galactans that are branches of rhamnogalacturonan I 1461:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 9 81 theme rhamnogalacturonan 1492:1509 arg1 branches 1480:1487 branches 1480:1487 branches of rhamnogalacturonan I 1480:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 9 82 with domain 1449:1454 arg1 galactans 1461:1469 galactans 1461:1469 galactans that are branches of rhamnogalacturonan I 1461:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 9 82 with domain 1449:1454 arg1 branches 1480:1487 branches 1480:1487 branches of rhamnogalacturonan I 1480:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 1 83 dep BACKGROUND 107:116 arg1 comprises 276:284 comprises 276:284 comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain 276:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 83 dep BACKGROUND 107:116 arg1 protein 143:149 Arabinogalactan protein 31 127:152 Arabinogalactan protein 31 (AGP31) 127:160 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 83 dep BACKGROUND 107:116 arg1 protein 194:200 a remarkable plant cell-wall protein 165:200 a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana 165:270 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 5 84 theme new 868:870 arg1 insight 872:878 new insight 868:878 new insight into its function in cell walls 868:910 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 2 85 from motifs 549:554 arg1 domain 572:577 the Pro-rich domain 559:577 the Pro-rich domain 559:577 AGP31 displays different O-glycosylation patterns with arabinogalactans on the AGP domain and Hyp-O-Gal/Ara-rich motifs on the Pro-rich domain. 24685714 7 86 theme recombinant 1253:1263 arg1 PAC-V5-6xHis 1265:1276 recombinant PAC-V5-6xHis 1253:1276 recombinant PAC-V5-6xHis 1253:1276 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 5 87 theme cell 901:904 arg1 walls 906:910 cell walls 901:910 cell walls 901:910 The purpose of the present study was to identify AGP31 partners to gain new insight into its function in cell walls. 24685714 15 88 theme walls 2198:2202 arg1 strengthening 2176:2188 the strengthening 2172:2188 the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls 2172:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 15 89 theme etiolated 2238:2246 arg1 hypocotyls 2248:2257 etiolated hypocotyls 2238:2257 etiolated hypocotyls 2238:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 10 90 theme entire 1590:1595 arg1 protein 1597:1603 an entire protein 1587:1603 an entire protein 1587:1603 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 1 91 theme signal 298:303 arg1 acids 353:357 seven amino acids 341:357 seven amino acids 341:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 peptide 305:311 a predicted signal peptide 286:311 a predicted signal peptide (SP) 286:316 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 domain 386:391 a Pro-rich domain 375:391 a Pro-rich domain 375:391 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 domain 331:336 a short AGP domain 319:336 a short AGP domain of seven amino acids 319:357 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 domain 428:433 a PAC (PRP-AGP containing Cys) domain 397:433 a PAC (PRP-AGP containing Cys) domain 397:433 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 His-stretch 362:372 a His-stretch 360:372 a His-stretch 360:372 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 1 91 theme signal 298:303 arg1 SP 314:315 SP 314:315 SP 314:315 BACKGROUND AND AIMS Arabinogalactan protein 31 (AGP31) is a remarkable plant cell-wall protein displaying a multi-domain organization unique in Arabidopsis thaliana: it comprises a predicted signal peptide (SP), a short AGP domain of seven amino acids, a His-stretch, a Pro-rich domain and a PAC (PRP-AGP containing Cys) domain. 24685714 7 92 theme purified 1228:1235 arg1 AGP31 1244:1248 native AGP31 1237:1248 native AGP31 1237:1248 After validation of the arrays, in vitro interaction assays were carried out by probing the membranes with purified native AGP31 or recombinant PAC-V5-6xHis. 24685714 9 93 dep RESULTS 1384:1390 arg1 demonstrated 1399:1410 demonstrated 1399:1410 was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I 1395:1511 KEY RESULTS It was demonstrated that AGP31 interacts through its PAC domain with galactans that are branches of rhamnogalacturonan I. 24685714 10 94 theme AGP31 1620:1624 arg1 homologues 1626:1635 AGP31 homologues 1620:1635 AGP31 homologues 1620:1635 This is the first experimental evidence that a PAC domain, also found as an entire protein or a domain of AGP31 homologues, can bind carbohydrates. 24685714 15 95 theme growing 2215:2221 arg1 hypocotyls 2248:2257 etiolated hypocotyls 2238:2257 etiolated hypocotyls 2238:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 15 95 theme growing 2215:2221 arg1 organs 2223:2228 quickly growing organs 2207:2228 quickly growing organs such as etiolated hypocotyls 2207:2257 Such scaffolds could contribute to the strengthening of cell walls of quickly growing organs such as etiolated hypocotyls. 24685714 8 96 theme scattering 1306:1315 arg1 analyses 1323:1330 dynamic light scattering (DLS) analyses 1292:1330 dynamic light scattering (DLS) analyses 1292:1330 In addition, dynamic light scattering (DLS) analyses were carried out on an AGP31 purified fraction. 24685714 4 97 theme Literature 721:730 arg1 data 732:735 Literature data 721:735 Literature data 721:735 Literature data suggest that AGP31 may interact with cell-wall components. 24685714 6 98 theme cell 1057:1060 arg1 walls 1062:1066 cell walls 1057:1066 cell walls of Arabidopsis thaliana and Brachypodium distachyon 1057:1118 METHODS Nitrocellulose membranes were prepared by spotting different polysaccharides, which were either obtained commercially or extracted from cell walls of Arabidopsis thaliana and Brachypodium distachyon. 24685714 14 99 with interactions 2012:2023 arg1 components 2059:2068 different cell-wall components 2039:2068 different cell-wall components 2039:2068 CONCLUSIONS These results allow the proposal of a model of interactions of AGP31 with different cell-wall components, in which AGP31 participates in complex supra-molecular scaffolds. 24105809 0 0 theme GCNT2 86:90 arg1 co-expression 58:70 co-expression 58:70 co-expression of β3GnT2 and GCNT2 58:90 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 9 1 theme high 1270:1273 arg1 levels 1275:1280 high levels 1270:1280 high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE 1270:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 5 2 theme N-glycan 703:710 arg1 branching 712:720 N-glycan branching 703:720 N-glycan branching 703:720 N-glycan branching and core composition, however, can also modulate the extent of PLN modification. 24105809 3 3 theme known 531:535 arg1 genes 543:547 eight known β3GnT genes 525:547 eight known β3GnT genes 525:547 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 9 4 theme PLN 1366:1368 arg1 glycans 1370:1376 PLN glycans 1366:1376 PLN glycans 1366:1376 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 10 5 theme PLN 1479:1481 arg1 levels 1483:1488 PLN levels 1479:1488 PLN levels in olfactory neurons 1479:1509 These data clearly suggest that GCNT2 functions in vivo together with β3GnT2 to determine PLN levels in olfactory neurons by regulating β1,6-branches that promote PLN extension. 24105809 3 6 theme β3GnT 537:541 arg1 genes 543:547 eight known β3GnT genes 525:547 eight known β3GnT genes 525:547 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 9 7 from cyclase 1336:1342 arg1 levels 1275:1280 high levels 1270:1280 high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE 1270:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 6 8 theme sensory 972:978 arg1 ganglia 980:986 the OE and other sensory ganglia 955:986 the OE and other sensory ganglia 955:986 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 3 9 from cells 441:445 arg1 ubiquitous 422:431 ubiquitous 422:431 ubiquitous 422:431 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 1 10 theme repeating 155:163 arg1 units 165:169 repeating units 155:169 repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 155:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 9 11 from co-transfection 1226:1240 arg1 cells 1253:1257 HEK293T cells 1245:1257 HEK293T cells 1245:1257 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 12 theme PLN 1285:1287 arg1 expression 1289:1298 PLN expression 1285:1298 PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE 1285:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 13 theme cell 1307:1310 arg1 surface 1312:1318 the cell surface 1303:1318 the cell surface 1303:1318 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 14 from β3GnT2 1209:1214 arg1 cells 1253:1257 HEK293T cells 1245:1257 HEK293T cells 1245:1257 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 15 theme expression 1289:1298 arg1 levels 1275:1280 high levels 1270:1280 high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE 1270:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 0 16 link N-linked 0:7 arg1 synthesis 32:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 4 17 theme axon 666:669 arg1 guidance 671:678 axon guidance 666:678 axon guidance 666:678 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 9 18 from levels 1275:1280 arg1 carrier 1355:1361 a major carrier 1347:1361 a major carrier of PLN glycans in the OE 1347:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 18 from levels 1275:1280 arg1 cyclase 1336:1342 adenylyl cyclase 3 1327:1344 adenylyl cyclase 3 1327:1344 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 18 from levels 1275:1280 arg1 surface 1312:1318 the cell surface 1303:1318 the cell surface 1303:1318 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 0 19 theme N-linked 0:7 arg1 synthesis 32:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 2 20 from expression 226:235 arg1 structures 265:274 glycoprotein core structures 247:274 glycoprotein core structures 247:274 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 10 21 theme olfactory 1493:1501 arg1 neurons 1503:1509 olfactory neurons 1493:1509 olfactory neurons 1493:1509 These data clearly suggest that GCNT2 functions in vivo together with β3GnT2 to determine PLN levels in olfactory neurons by regulating β1,6-branches that promote PLN extension. 24105809 0 22 theme polylactosamine 9:23 arg1 synthesis 32:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 10 23 theme PLN 1552:1554 arg1 extension 1556:1564 PLN extension 1552:1564 PLN extension 1552:1564 These data clearly suggest that GCNT2 functions in vivo together with β3GnT2 to determine PLN levels in olfactory neurons by regulating β1,6-branches that promote PLN extension. 24105809 3 24 from transcription 508:520 arg1 mice 552:555 mice 552:555 mice 552:555 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 2 25 gly glycoprotein 247:258 arg1 glycoprotein 247:258 glycoprotein core structures 247:274 glycoprotein core structures 247:274 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 6 26 theme OE 959:960 arg1 ganglia 980:986 the OE and other sensory ganglia 955:986 the OE and other sensory ganglia 955:986 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 9 27 from surface 1312:1318 arg1 levels 1275:1280 high levels 1270:1280 high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE 1270:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 28 theme this 1203:1206 arg1 support 1192:1198 support 1192:1198 support of this 1192:1206 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 3 29 theme PLN 448:450 arg1 expression 452:461 PLN expression 448:461 PLN expression 448:461 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 4 30 theme olfactory 565:573 arg1 epithelium 575:584 the olfactory epithelium 561:584 the olfactory epithelium (OE) 561:589 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 4 30 theme olfactory 565:573 arg1 OE 587:588 OE 587:588 OE 587:588 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 1 31 theme disaccharide 185:196 arg1 n 218:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 0 32 theme glycan 25:30 arg1 synthesis 32:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis 0:40 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 7 33 theme olfactory 1032:1040 arg1 neurons 1042:1048 mature olfactory neurons 1025:1048 mature olfactory neurons that co-express β3GnT2 and PLN 1025:1079 Postnatally, GCNT2 is maintained in mature olfactory neurons that co-express β3GnT2 and PLN. 24105809 6 34 theme glycosyltransferase 860:878 arg1 GCNT2 880:884 the β1,6-branching glycosyltransferase GCNT2 841:884 the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) 841:909 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 2 35 theme enzyme 295:300 arg1 activities 302:311 enzyme activities 295:311 enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT) 295:399 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 4 36 theme chains 636:641 arg1 expression 609:618 expression 609:618 expression of extended PLN chains that are essential for axon guidance and neuronal survival 609:700 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 3 37 theme most 436:439 arg1 cells 441:445 most cells 436:445 most cells 436:445 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 5 38 theme core 726:729 arg1 composition 731:741 core composition 726:741 core composition 726:741 N-glycan branching and core composition, however, can also modulate the extent of PLN modification. 24105809 6 39 theme β1,6-branching 845:858 arg1 GCNT2 880:884 the β1,6-branching glycosyltransferase GCNT2 841:884 the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) 841:909 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 1 40 theme common 178:183 arg1 n 218:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 1 41 theme Galβ1,4-GlcNAcβ1,3 199:216 arg1 n 218:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 174:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 5 42 theme PLN 785:787 arg1 modification 789:800 PLN modification 785:800 PLN modification 785:800 N-glycan branching and core composition, however, can also modulate the extent of PLN modification. 24105809 9 43 theme adenylyl 1327:1334 arg1 carrier 1355:1361 a major carrier 1347:1361 a major carrier of PLN glycans in the OE 1347:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 43 theme adenylyl 1327:1334 arg1 cyclase 1336:1342 adenylyl cyclase 3 1327:1344 adenylyl cyclase 3 1327:1344 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 44 theme glycans 1370:1376 arg1 carrier 1355:1361 a major carrier 1347:1361 a major carrier of PLN glycans in the OE 1347:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 44 theme glycans 1370:1376 arg1 cyclase 1336:1342 adenylyl cyclase 3 1327:1344 adenylyl cyclase 3 1327:1344 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 45 theme GCNT2 1220:1224 arg1 co-transfection 1226:1240 GCNT2 co-transfection 1220:1240 GCNT2 co-transfection 1220:1240 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 6 46 dep known 896:900 arg1 formerly 887:894 formerly 887:894 formerly 887:894 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 5 47 theme modification 789:800 arg1 extent 775:780 the extent 771:780 the extent of PLN modification 771:800 N-glycan branching and core composition, however, can also modulate the extent of PLN modification. 24105809 6 48 theme high 927:930 arg1 levels 932:937 high levels 927:937 high levels 927:937 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 9 49 from carrier 1355:1361 arg1 OE 1385:1386 the OE 1381:1386 the OE 1381:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 10 50 from levels 1483:1488 arg1 neurons 1503:1509 olfactory neurons 1493:1509 olfactory neurons 1493:1509 These data clearly suggest that GCNT2 functions in vivo together with β3GnT2 to determine PLN levels in olfactory neurons by regulating β1,6-branches that promote PLN extension. 24105809 7 51 theme mature 1025:1030 arg1 neurons 1042:1048 mature olfactory neurons 1025:1048 mature olfactory neurons that co-express β3GnT2 and PLN 1025:1079 Postnatally, GCNT2 is maintained in mature olfactory neurons that co-express β3GnT2 and PLN. 24105809 9 52 theme HEK293T 1245:1251 arg1 cells 1253:1257 HEK293T cells 1245:1257 HEK293T cells 1245:1257 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 6 53 theme first 825:829 arg1 time 831:834 the first time 821:834 the first time 821:834 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 2 54 theme glycoprotein 247:258 arg1 structures 265:274 glycoprotein core structures 247:274 glycoprotein core structures 247:274 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 8 55 theme specific 1094:1101 arg1 co-expression 1103:1115 This highly specific co-expression 1082:1115 This highly specific co-expression 1082:1115 This highly specific co-expression suggests that GCNT2 and β3GnT2 function cooperatively in PLN synthesis. 24105809 3 56 theme tissue-specific 492:506 arg1 transcription 508:520 the tissue-specific transcription 488:520 the tissue-specific transcription of eight known β3GnT genes in mice 488:555 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 1 57 theme n 218:218 arg1 units 165:169 repeating units 155:169 repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 155:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 2 58 theme core 260:263 arg1 structures 265:274 glycoprotein core structures 247:274 glycoprotein core structures 247:274 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 9 59 theme major 1349:1353 arg1 carrier 1355:1361 a major carrier 1347:1361 a major carrier of PLN glycans in the OE 1347:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 59 theme major 1349:1353 arg1 cyclase 1336:1342 adenylyl cyclase 3 1327:1344 adenylyl cyclase 3 1327:1344 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 8 60 theme PLN 1174:1176 arg1 synthesis 1178:1186 PLN synthesis 1174:1186 PLN synthesis 1174:1186 This highly specific co-expression suggests that GCNT2 and β3GnT2 function cooperatively in PLN synthesis. 24105809 1 61 theme unique 129:134 arg1 Poly-N-acetyllactosamine 93:116 Poly-N-acetyllactosamine 93:116 Poly-N-acetyllactosamine (PLN) 93:122 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 1 61 theme unique 129:134 arg1 glycan 136:141 a unique glycan 127:141 a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n 127:218 Poly-N-acetyllactosamine (PLN) is a unique glycan composed of repeating units of the common disaccharide (Galβ1,4-GlcNAcβ1,3)n . 24105809 6 62 dep GCNT2 880:884 arg1 known 896:900 known 896:900 known as IGnT 896:908 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 0 63 theme β3GnT2 75:80 arg1 co-expression 58:70 co-expression 58:70 co-expression of β3GnT2 and GCNT2 58:90 N-linked polylactosamine glycan synthesis is regulated by co-expression of β3GnT2 and GCNT2. 24105809 2 64 theme PLN 240:242 arg1 expression 226:235 The expression 222:235 The expression of PLN on glycoprotein core structures 222:274 The expression of PLN on glycoprotein core structures minimally requires enzyme activities for β1,4-galactosyltransferase (β4GalT) and β1,3-N-acetylglucosminyltransferase (β3GnT). 24105809 3 65 theme genes 543:547 arg1 transcription 508:520 the tissue-specific transcription 488:520 the tissue-specific transcription of eight known β3GnT genes in mice 488:555 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 4 66 theme PLN 632:634 arg1 chains 636:641 extended PLN chains 623:641 extended PLN chains that are essential for axon guidance and neuronal survival 623:700 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 6 67 theme other 966:970 arg1 ganglia 980:986 the OE and other sensory ganglia 955:986 the OE and other sensory ganglia 955:986 Here, we show for the first time that the β1,6-branching glycosyltransferase GCNT2 (formerly known as IGnT) is expressed at high levels specifically in the OE and other sensory ganglia. 24105809 3 68 from ubiquitous 422:431 arg1 cells 441:445 most cells 436:445 most cells 436:445 Because β4GalTs are ubiquitous in most cells, PLN expression is generally ascribed to the tissue-specific transcription of eight known β3GnT genes in mice. 24105809 9 69 from expression 1289:1298 arg1 carrier 1355:1361 a major carrier 1347:1361 a major carrier of PLN glycans in the OE 1347:1386 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 69 from expression 1289:1298 arg1 cyclase 1336:1342 adenylyl cyclase 3 1327:1344 adenylyl cyclase 3 1327:1344 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 9 69 from expression 1289:1298 arg1 surface 1312:1318 the cell surface 1303:1318 the cell surface 1303:1318 In support of this, β3GnT2 and GCNT2 co-transfection in HEK293T cells results in high levels of PLN expression on the cell surface and on adenylyl cyclase 3, a major carrier of PLN glycans in the OE. 24105809 4 70 theme neuronal 684:691 arg1 survival 693:700 neuronal survival 684:700 neuronal survival 684:700 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 24105809 4 71 theme extended 623:630 arg1 chains 636:641 extended PLN chains 623:641 extended PLN chains that are essential for axon guidance and neuronal survival 623:700 In the olfactory epithelium (OE), β3GnT2 regulates expression of extended PLN chains that are essential for axon guidance and neuronal survival. 26416237 3 0 theme aspersa 295:301 arg1 HAA 315:317 HAA 315:317 HAA 315:317 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 0 theme aspersa 295:301 arg1 agglutinin 303:312 Helix aspersa agglutinin 289:312 Helix aspersa agglutinin (HAA) 289:318 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 0 theme aspersa 295:301 arg1 lectin 325:330 a lectin 323:330 a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells 323:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 7 1 theme glycan 1049:1054 arg1 chain 1056:1060 the HAA natural glycan chain 1033:1060 the HAA natural glycan chain 1033:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 4 2 theme diagnostic 536:545 arg1 tool 547:550 a diagnostic tool 534:550 a diagnostic tool 534:550 Although several reports have already described the use of HAA as a diagnostic tool, this protein was not characterized on the molecular level. 26416237 5 3 theme Helix 711:715 arg1 aspersa 717:723 Helix aspersa 711:723 Helix aspersa (garden snail) 711:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 5 3 theme Helix 711:715 arg1 snail 733:737 garden snail 726:737 garden snail 726:737 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 8 4 with topology 1067:1074 arg1 lectin 1125:1130 lectin 1125:1130 lectin 1125:1130 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 7 5 theme chain 1056:1060 arg1 part 1025:1028 a large part 1017:1028 a large part of the HAA natural glycan chain 1017:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 6 6 theme amino 745:749 arg1 sequence 756:763 The amino acid sequence 741:763 The amino acid sequence of this agglutinin 741:782 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 1 7 theme differentiated 133:146 arg1 proteins 157:164 proteins 157:164 proteins known to possess sugar-binding properties 157:206 Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. 26416237 1 7 theme differentiated 133:146 arg1 group 148:152 a differentiated group 131:152 a differentiated group of proteins known to possess sugar-binding properties 131:206 Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. 26416237 9 8 theme important 1346:1354 arg1 tool 1367:1370 this important diagnostic tool 1341:1370 this important diagnostic tool 1341:1370 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 8 9 theme GalNAc 1083:1088 arg1 cleft 1098:1102 the GalNAc binding cleft 1079:1102 the GalNAc binding cleft 1079:1102 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 4 10 theme several 477:483 arg1 reports 485:491 several reports 477:491 several reports 477:491 Although several reports have already described the use of HAA as a diagnostic tool, this protein was not characterized on the molecular level. 26416237 7 11 theme HAA 1037:1039 arg1 chain 1056:1060 the HAA natural glycan chain 1033:1060 the HAA natural glycan chain 1033:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 3 12 contain containing 361:370 arg2 GalNAc 399:404 GalNAc 399:404 GalNAc 399:404 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 12 contain containing 361:370 arg2 α-d-N-acetylgalactosamine 372:396 α-d-N-acetylgalactosamine 372:396 α-d-N-acetylgalactosamine (GalNAc) 372:405 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 12 contain containing 361:370 arg1 epitopes 352:359 the epitopes 348:359 the epitopes 348:359 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 12 contain containing 361:370 arg1 present 417:423 present 417:423 present 417:423 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 9 13 theme diagnostic 1356:1365 arg1 tool 1367:1370 this important diagnostic tool 1341:1370 this important diagnostic tool 1341:1370 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 7 14 theme natural 1041:1047 arg1 chain 1056:1060 the HAA natural glycan chain 1033:1060 the HAA natural glycan chain 1033:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 8 15 theme HSQC 1198:1201 arg1 spectroscopy 1207:1218 STD HSQC NMR spectroscopy 1194:1218 STD HSQC NMR spectroscopy 1194:1218 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 9 16 theme tool 1367:1370 arg1 modifications 1324:1336 rational modifications 1315:1336 rational modifications of this important diagnostic tool 1315:1370 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 8 17 theme cleft 1098:1102 arg1 topology 1067:1074 The topology 1063:1074 The topology of the GalNAc binding cleft 1063:1102 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 8 17 theme cleft 1098:1102 arg1 interaction 1108:1118 interaction 1108:1118 interaction with lectin 1108:1130 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 8 18 theme NMR 1203:1205 arg1 spectroscopy 1207:1218 STD HSQC NMR spectroscopy 1194:1218 STD HSQC NMR spectroscopy 1194:1218 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 3 19 located present 417:423 arg2 epitopes 352:359 the epitopes 348:359 the epitopes 348:359 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 19 located present 417:423 arg2 present 417:423 present 417:423 present 417:423 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 19 located present 417:423 arg1 surface 432:438 the surface 428:438 the surface of metastatic cancer cells 428:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 2 20 theme research 248:255 arg1 targets 257:263 interesting research targets 236:263 interesting research targets in medical diagnostics 236:286 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 2 20 theme research 248:255 arg1 they 227:230 they 227:230 they 227:230 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 8 21 theme binding 1090:1096 arg1 cleft 1098:1102 the GalNAc binding cleft 1079:1102 the GalNAc binding cleft 1079:1102 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 5 22 theme aspersa 717:723 arg1 mucus 702:706 mucus 702:706 mucus of Helix aspersa (garden snail) 702:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 2 23 theme interesting 236:246 arg1 targets 257:263 interesting research targets 236:263 interesting research targets in medical diagnostics 236:286 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 2 23 theme interesting 236:246 arg1 they 227:230 they 227:230 they 227:230 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 7 24 theme resolution 900:909 arg1 1.38Å 930:934 1.38Å 930:934 1.38Å 930:934 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 7 24 theme resolution 900:909 arg1 structure 919:927 The high resolution crystal structure 891:927 The high resolution crystal structure (1.38Å) 891:935 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 6 25 theme Edman 802:806 arg1 degradation 808:818 Edman degradation 802:818 Edman degradation 802:818 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 7 26 theme large 1019:1023 arg1 part 1025:1028 a large part 1017:1028 a large part of the HAA natural glycan chain 1017:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 1 27 theme proteins 157:164 arg1 proteins 157:164 proteins 157:164 proteins known to possess sugar-binding properties 157:206 Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. 26416237 1 27 theme proteins 157:164 arg1 group 148:152 a differentiated group 131:152 a differentiated group of proteins known to possess sugar-binding properties 131:206 Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. 26416237 0 28 theme Structural 0:9 arg1 studies 11:17 Structural studies 0:17 Structural studies of Helix aspersa agglutinin 0:45 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 9 29 theme structural 1245:1254 arg1 clues 1256:1260 structural clues 1245:1260 structural clues regarding HAA specificity 1245:1286 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 0 30 theme Helix 22:26 arg1 agglutinin 36:45 Helix aspersa agglutinin 22:45 Helix aspersa agglutinin 22:45 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 5 31 theme first 637:641 arg1 time 643:646 the first time 633:646 the first time 633:646 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 3 32 attach present 417:423 arg2 epitopes 352:359 the epitopes 348:359 the epitopes 348:359 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 32 attach present 417:423 arg2 present 417:423 present 417:423 present 417:423 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 32 attach present 417:423 arg1 surface 432:438 the surface 428:438 the surface of metastatic cancer cells 428:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 7 33 theme high 895:898 arg1 1.38Å 930:934 1.38Å 930:934 1.38Å 930:934 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 7 33 theme high 895:898 arg1 structure 919:927 The high resolution crystal structure 891:927 The high resolution crystal structure (1.38Å) 891:935 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 0 34 theme diagnostic 96:105 arg1 lectin 72:77 A lectin 70:77 A lectin that serves as a diagnostic tool 70:110 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 0 34 theme diagnostic 96:105 arg1 tool 107:110 a diagnostic tool 94:110 a diagnostic tool 94:110 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 2 35 theme medical 268:274 arg1 diagnostics 276:286 medical diagnostics 268:286 medical diagnostics 268:286 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 0 36 theme agglutinin 36:45 arg1 studies 11:17 Structural studies 0:17 Structural studies of Helix aspersa agglutinin 0:45 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 4 37 theme HAA 527:529 arg1 use 520:522 the use 516:522 the use of HAA as a diagnostic tool 516:550 Although several reports have already described the use of HAA as a diagnostic tool, this protein was not characterized on the molecular level. 26416237 7 38 theme detailed 990:997 arg1 information 999:1009 the detailed information 986:1009 the detailed information about a large part of the HAA natural glycan chain 986:1060 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 4 39 theme molecular 595:603 arg1 level 605:609 the molecular level 591:609 the molecular level 591:609 Although several reports have already described the use of HAA as a diagnostic tool, this protein was not characterized on the molecular level. 26416237 1 40 theme sugar-binding 183:195 arg1 properties 197:206 sugar-binding properties 183:206 sugar-binding properties 183:206 Lectins belong to a differentiated group of proteins known to possess sugar-binding properties. 26416237 0 41 theme aspersa 28:34 arg1 agglutinin 36:45 Helix aspersa agglutinin 22:45 Helix aspersa agglutinin 22:45 Structural studies of Helix aspersa agglutinin complexed with GalNAc: A lectin that serves as a diagnostic tool. 26416237 5 42 theme structural 652:661 arg1 information 663:673 the structural information 648:673 the structural information about lectin isolated from mucus of Helix aspersa (garden snail) 648:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 8 43 theme STD 1194:1196 arg1 spectroscopy 1207:1218 STD HSQC NMR spectroscopy 1194:1218 STD HSQC NMR spectroscopy 1194:1218 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 7 44 theme MALDI-TOF 941:949 arg1 spectrometry 956:967 MALDI-TOF mass spectrometry 941:967 MALDI-TOF mass spectrometry analysis 941:976 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 5 45 theme garden 726:731 arg1 aspersa 717:723 Helix aspersa 711:723 Helix aspersa (garden snail) 711:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 5 45 theme garden 726:731 arg1 snail 733:737 garden snail 726:737 garden snail 726:737 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 6 46 theme X-ray 868:872 arg1 crystallography 874:888 X-ray crystallography 868:888 X-ray crystallography 868:888 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 6 47 theme agglutinin 773:782 arg1 sequence 756:763 The amino acid sequence 741:763 The amino acid sequence of this agglutinin 741:782 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 7 48 theme mass 951:954 arg1 spectrometry 956:967 MALDI-TOF mass spectrometry 941:967 MALDI-TOF mass spectrometry analysis 941:976 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 3 49 theme cancer 454:459 arg1 cells 461:465 metastatic cancer cells 443:465 metastatic cancer cells 443:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 50 theme Helix 289:293 arg1 HAA 315:317 HAA 315:317 HAA 315:317 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 50 theme Helix 289:293 arg1 agglutinin 303:312 Helix aspersa agglutinin 289:312 Helix aspersa agglutinin (HAA) 289:318 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 50 theme Helix 289:293 arg1 lectin 325:330 a lectin 323:330 a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells 323:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 8 51 with interaction 1108:1118 arg1 lectin 1125:1130 lectin 1125:1130 lectin 1125:1130 The topology of the GalNAc binding cleft and interaction with lectin are very well defined in the structure and fully confirmed by STD HSQC NMR spectroscopy. 26416237 3 52 theme cells 461:465 arg1 surface 432:438 the surface 428:438 the surface of metastatic cancer cells 428:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 7 53 theme spectrometry 956:967 arg1 analysis 969:976 MALDI-TOF mass spectrometry analysis 941:976 MALDI-TOF mass spectrometry analysis 941:976 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 9 54 theme HAA 1272:1274 arg1 specificity 1276:1286 HAA specificity 1272:1286 HAA specificity 1272:1286 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 3 55 from surface 432:438 arg1 epitopes 352:359 the epitopes 348:359 the epitopes 348:359 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 3 55 from surface 432:438 arg1 present 417:423 present 417:423 present 417:423 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 7 56 theme crystal 911:917 arg1 1.38Å 930:934 1.38Å 930:934 1.38Å 930:934 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 7 56 theme crystal 911:917 arg1 structure 919:927 The high resolution crystal structure 891:927 The high resolution crystal structure (1.38Å) 891:935 The high resolution crystal structure (1.38Å) and MALDI-TOF mass spectrometry analysis provide the detailed information about a large part of the HAA natural glycan chain. 26416237 3 57 theme metastatic 443:452 arg1 cells 461:465 metastatic cancer cells 443:465 metastatic cancer cells 443:465 Helix aspersa agglutinin (HAA) is a lectin that recognizes the epitopes containing α-d-N-acetylgalactosamine (GalNAc), which is present at the surface of metastatic cancer cells. 26416237 9 58 theme rational 1315:1322 arg1 modifications 1324:1336 rational modifications 1315:1336 rational modifications of this important diagnostic tool 1315:1370 Together, this provides structural clues regarding HAA specificity and opens possibilities to rational modifications of this important diagnostic tool. 26416237 6 59 theme acid 751:754 arg1 sequence 756:763 The amino acid sequence 741:763 The amino acid sequence of this agglutinin 741:782 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 2 60 from targets 257:263 arg1 diagnostics 276:286 medical diagnostics 268:286 medical diagnostics 268:286 Due to this fact, they are interesting research targets in medical diagnostics. 26416237 6 61 theme quaternary 844:853 arg1 structure 855:863 quaternary structure 844:863 Edman degradation and tertiary as well as quaternary structure 802:863 The amino acid sequence of this agglutinin was determined by Edman degradation and tertiary as well as quaternary structure by X-ray crystallography. 26416237 5 62 attach isolated 688:695 arg1 mucus 702:706 mucus 702:706 mucus of Helix aspersa (garden snail) 702:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 26416237 5 62 attach isolated 688:695 arg2 lectin 681:686 lectin 681:686 lectin isolated from mucus of Helix aspersa (garden snail) 681:738 Here, we present for the first time the structural information about lectin isolated from mucus of Helix aspersa (garden snail). 24403418 4 0 theme hERG 543:546 arg1 channel 583:589 hERG (human ether-a-go-go related gene) channel 543:589 hERG (human ether-a-go-go related gene) channel 543:589 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 4 0 theme hERG 543:546 arg1 gene 577:580 human ether-a-go-go related gene 549:580 human ether-a-go-go related gene 549:580 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 9 1 theme repolarization 1351:1364 arg1 phase 1366:1370 the repolarization phase 1347:1370 the repolarization phase of cardiac APs 1347:1385 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 4 2 theme human 549:553 arg1 channel 583:589 hERG (human ether-a-go-go related gene) channel 543:589 hERG (human ether-a-go-go related gene) channel 543:589 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 4 2 theme human 549:553 arg1 gene 577:580 human ether-a-go-go related gene 549:580 human ether-a-go-go related gene 549:580 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 1 3 theme action 117:122 arg1 AP 136:137 AP 136:137 AP 136:137 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 1 3 theme action 117:122 arg1 potentials 124:133 Cardiac action potentials 109:133 Cardiac action potentials (AP) 109:138 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 10 4 theme electrocardiogram 1499:1515 arg1 patterns 1479:1486 the patterns 1475:1486 the patterns of derived electrocardiogram 1475:1515 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 1 5 theme orchestrated 160:171 arg1 functions 173:181 the orchestrated functions 156:181 the orchestrated functions of ion channels 156:197 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 8 6 theme cardiac 1215:1221 arg1 processes 1223:1231 multiscale cardiac processes 1204:1231 multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues) 1204:1274 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 10 7 theme hERG 1552:1555 arg1 channel 1557:1563 hERG channel 1552:1563 hERG channel 1552:1563 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 6 8 theme Ovary 911:915 arg1 cells 923:927 Chinese Hamster Ovary (CHO) cells 895:927 Chinese Hamster Ovary (CHO) cells 895:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 5 9 theme modulation 771:780 arg1 dynamics 782:789 the glycosylation modulation dynamics 753:789 the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals 753:841 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 6 10 theme Chinese 895:901 arg1 cells 923:927 Chinese Hamster Ovary (CHO) cells 895:927 Chinese Hamster Ovary (CHO) cells 895:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 10 11 theme QT 1578:1579 arg1 interval 1581:1588 the QT interval 1574:1588 the QT interval 1574:1588 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 5 12 from dynamics 782:789 arg1 signals 835:841 cardiac electrical signals 816:841 cardiac electrical signals 816:841 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 5 12 from dynamics 782:789 arg1 channels 803:810 hERG ion channels 794:810 hERG ion channels 794:810 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 8 13 theme reduced 1179:1185 arg1 glycosylation 1187:1199 reduced glycosylation 1179:1199 reduced glycosylation 1179:1199 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 2 14 theme slight 202:207 arg1 change 209:214 A slight change 200:214 A slight change in ion channel activity 200:238 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 9 15 theme electrical 1408:1417 arg1 propagation 1419:1429 electrical propagation 1408:1429 electrical propagation in cardiac fibers and tissues 1408:1459 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 6 16 theme hERG 868:871 arg1 channels 873:880 hERG channels 868:880 hERG channels expressed in Chinese Hamster Ovary (CHO) cells 868:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 4 17 theme AP 659:660 arg1 phase 632:636 late phase 2 627:638 late phase 2 627:638 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 4 17 theme AP 659:660 arg1 phase 644:648 phase 3 644:650 phase 3 644:650 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 11 18 theme QT 1698:1699 arg1 syndrome 1701:1708 the long QT syndrome 1689:1708 the long QT syndrome 1689:1708 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 3 19 theme mass 427:430 arg1 % 403:403 up to 30% 395:403 up to 30% of a mature protein's mass comprised of glycan structures 395:461 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 3 19 theme mass 427:430 arg1 mass 427:430 a mature protein's mass 408:430 a mature protein's mass comprised of glycan structures 408:461 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 8 20 theme 2-D 1263:1265 arg1 tissues 1267:1273 2-D tissues 1263:1273 2-D tissues 1263:1273 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 0 21 theme ion 64:66 arg1 channels 68:75 hERG ion channels 59:75 hERG ion channels 59:75 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 2 22 theme channel 223:229 arg1 activity 231:238 ion channel activity 219:238 ion channel activity 219:238 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 5 23 theme electrical 824:833 arg1 signals 835:841 cardiac electrical signals 816:841 cardiac electrical signals 816:841 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 11 24 theme other 1726:1730 arg1 disorders 1740:1748 potentially other cardiac disorders 1714:1748 potentially other cardiac disorders 1714:1748 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 5 25 theme in 699:700 arg1 experiments 708:718 in vitro experiments 699:718 in vitro experiments with in-silico models 699:740 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 3 26 theme ion 351:353 arg1 channels 355:362 Cardiac ion channels 343:362 Cardiac ion channels 343:362 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 0 27 theme electrical 89:98 arg1 signals 100:106 cardiac electrical signals 81:106 cardiac electrical signals 81:106 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 2 28 from change 209:214 arg1 activity 231:238 ion channel activity 219:238 ion channel activity 219:238 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 6 29 theme CHO 918:920 arg1 cells 923:927 Chinese Hamster Ovary (CHO) cells 895:927 Chinese Hamster Ovary (CHO) cells 895:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 10 30 theme re-entry 1608:1615 arg1 rate 1617:1620 the re-entry rate 1604:1620 the re-entry rate of spiral waves 1604:1636 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 3 31 gly glycosylated 376:387 arg1 channels 355:362 Cardiac ion channels 343:362 Cardiac ion channels 343:362 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 8 32 dep processes 1223:1231 arg1 1-D 1255:1257 1-D 1255:1257 1-D 1255:1257 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 8 32 dep processes 1223:1231 arg1 tissues 1267:1273 2-D tissues 1263:1273 2-D tissues 1263:1273 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 8 32 dep processes 1223:1231 arg1 cells 1248:1252 cardiac cells 1240:1252 cardiac cells 1240:1252 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 6 33 dep glycosylation 991:1003 arg1 i.e. 980:983 i.e. 980:983 i.e. 980:983 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 0 34 theme In-silico 0:8 arg1 modeling 10:17 In-silico modeling 0:17 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 0:107 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 6 35 theme full 986:989 arg1 glycosylation 991:1003 full glycosylation 986:1003 full glycosylation 986:1003 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 10 36 theme waves 1632:1636 arg1 rate 1617:1620 the re-entry rate 1604:1620 the re-entry rate of spiral waves 1604:1636 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 1 37 theme Cardiac 109:115 arg1 AP 136:137 AP 136:137 AP 136:137 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 1 37 theme Cardiac 109:115 arg1 potentials 124:133 Cardiac action potentials 109:133 Cardiac action potentials (AP) 109:138 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 9 38 theme cardiac 1375:1381 arg1 APs 1383:1385 cardiac APs 1375:1385 cardiac APs 1375:1385 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 0 39 theme modulation 36:45 arg1 dynamics 47:54 glycosylation modulation dynamics 22:54 glycosylation modulation dynamics 22:54 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 9 40 from propagation 1419:1429 arg1 tissues 1453:1459 tissues 1453:1459 tissues 1453:1459 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 9 40 from propagation 1419:1429 arg1 fibers 1442:1447 cardiac fibers 1434:1447 cardiac fibers 1434:1447 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 9 41 theme APs 1383:1385 arg1 phase 1366:1370 the repolarization phase 1347:1370 the repolarization phase of cardiac APs 1347:1385 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 5 42 theme in-silico 725:733 arg1 models 735:740 in-silico models 725:740 in-silico models 725:740 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 9 43 theme reduced 1304:1310 arg1 glycosylation 1312:1324 reduced glycosylation 1304:1324 reduced glycosylation 1304:1324 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 2 44 theme abnormal 317:324 arg1 rhythms 334:340 abnormal cardiac rhythms 317:340 abnormal cardiac rhythms 317:340 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 4 45 theme ether-a-go-go 555:567 arg1 channel 583:589 hERG (human ether-a-go-go related gene) channel 543:589 hERG (human ether-a-go-go related gene) channel 543:589 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 4 45 theme ether-a-go-go 555:567 arg1 gene 577:580 human ether-a-go-go related gene 549:580 human ether-a-go-go related gene 549:580 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 10 46 theme derived 1491:1497 arg1 electrocardiogram 1499:1515 derived electrocardiogram 1491:1515 derived electrocardiogram 1491:1515 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 11 47 theme pharmaceutical 1662:1675 arg1 targets 1677:1683 new pharmaceutical targets 1658:1683 new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders 1658:1748 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 5 48 theme ion 799:801 arg1 channels 803:810 hERG ion channels 794:810 hERG ion channels 794:810 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 6 49 theme gating 848:853 arg1 behaviors 855:863 The gating behaviors 844:863 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells 844:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 10 50 theme reduced 1527:1533 arg1 glycosylation 1535:1547 reduced glycosylation 1527:1547 reduced glycosylation of hERG channel 1527:1563 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 8 51 theme in-silico 1088:1096 arg1 models 1098:1103 in-silico models 1088:1103 in-silico models 1088:1103 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 2 52 theme AP 255:256 arg1 waveform 258:265 the AP waveform 251:265 the AP waveform 251:265 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 4 53 theme late 627:630 arg1 phase 632:636 late phase 2 627:638 late phase 2 627:638 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 5 54 theme hERG 794:797 arg1 channels 803:810 hERG ion channels 794:810 hERG ion channels 794:810 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 8 55 from effects 1168:1174 arg1 processes 1223:1231 multiscale cardiac processes 1204:1231 multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues) 1204:1274 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 5 56 with experiments 708:718 arg1 models 735:740 in-silico models 725:740 in-silico models 725:740 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 8 57 theme glycosylation 1187:1199 arg1 effects 1168:1174 the effects 1164:1174 the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues) 1164:1274 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 10 58 theme channel 1557:1563 arg1 glycosylation 1535:1547 reduced glycosylation 1527:1547 reduced glycosylation of hERG channel 1527:1563 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 8 59 theme glycosylation-channel 1117:1137 arg1 interactions 1139:1150 glycosylation-channel interactions 1117:1150 glycosylation-channel interactions 1117:1150 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 8 60 theme multiscale 1204:1213 arg1 processes 1223:1231 multiscale cardiac processes 1204:1231 multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues) 1204:1274 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 5 61 theme glycosylation 757:769 arg1 dynamics 782:789 the glycosylation modulation dynamics 753:789 the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals 753:841 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 6 62 theme Hamster 903:909 arg1 cells 923:927 Chinese Hamster Ovary (CHO) cells 895:927 Chinese Hamster Ovary (CHO) cells 895:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 5 63 dep in 699:700 arg1 vitro 702:706 vitro 702:706 vitro 702:706 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 4 64 theme channel 583:589 arg1 gating 533:538 the gating 529:538 the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP 529:660 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 2 65 theme ion 219:221 arg1 activity 231:238 ion channel activity 219:238 ion channel activity 219:238 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 9 66 theme cardiac 1434:1440 arg1 fibers 1442:1447 cardiac fibers 1434:1447 cardiac fibers 1434:1447 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 6 67 theme channels 873:880 arg1 behaviors 855:863 The gating behaviors 844:863 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells 844:927 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 8 68 theme cardiac 1240:1246 arg1 cells 1248:1252 cardiac cells 1240:1252 cardiac cells 1240:1252 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 24403418 6 69 theme glycosylation 954:966 arg1 mannose-rich 1027:1038 mannose-rich 1027:1038 mannose-rich 1027:1038 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 6 69 theme glycosylation 954:966 arg1 conditions 968:977 four glycosylation conditions 949:977 four glycosylation conditions 949:977 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 6 69 theme glycosylation 954:966 arg1 sialylation 1014:1024 reduced sialylation 1006:1024 reduced sialylation 1006:1024 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 1 70 theme ion 186:188 arg1 channels 190:197 ion channels 186:197 ion channels 186:197 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 5 71 theme cardiac 816:822 arg1 signals 835:841 cardiac electrical signals 816:841 cardiac electrical signals 816:841 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 11 72 theme long 1693:1696 arg1 syndrome 1701:1708 the long QT syndrome 1689:1708 the long QT syndrome 1689:1708 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 0 73 theme cardiac 81:87 arg1 signals 100:106 cardiac electrical signals 81:106 cardiac electrical signals 81:106 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 0 74 from modeling 10:17 arg1 signals 100:106 cardiac electrical signals 81:106 cardiac electrical signals 81:106 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 0 74 from modeling 10:17 arg1 channels 68:75 hERG ion channels 59:75 hERG ion channels 59:75 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 11 75 theme cardiac 1732:1738 arg1 disorders 1740:1748 potentially other cardiac disorders 1714:1748 potentially other cardiac disorders 1714:1748 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 5 76 from experiments 708:718 arg1 data 689:692 the data 685:692 the data from in vitro experiments with in-silico models 685:740 This paper integrates the data from in vitro experiments with in-silico models to predict the glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 1 77 theme channels 190:197 arg1 functions 173:181 the orchestrated functions 156:181 the orchestrated functions of ion channels 156:197 Cardiac action potentials (AP) are produced by the orchestrated functions of ion channels. 24403418 6 78 theme reduced 1006:1012 arg1 sialylation 1014:1024 reduced sialylation 1006:1024 reduced sialylation 1006:1024 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 6 78 theme reduced 1006:1012 arg1 conditions 968:977 four glycosylation conditions 949:977 four glycosylation conditions 949:977 The gating behaviors of hERG channels expressed in Chinese Hamster Ovary (CHO) cells were measured under four glycosylation conditions, i.e., full glycosylation, reduced sialylation, mannose-rich. 24403418 3 79 theme Cardiac 343:349 arg1 channels 355:362 Cardiac ion channels 343:362 Cardiac ion channels 343:362 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 3 80 theme mature 410:415 arg1 protein 417:423 a mature protein 408:423 a mature protein's mass comprised of glycan structures 408:461 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 3 81 dep 30 401:402 arg1 to 398:399 to 398:399 to 398:399 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 10 82 gly glycosylation 1535:1547 arg1 channel 1557:1563 hERG channel 1552:1563 hERG channel 1552:1563 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 0 83 theme glycosylation 22:34 arg1 dynamics 47:54 glycosylation modulation dynamics 22:54 glycosylation modulation dynamics 22:54 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 10 84 theme spiral 1625:1630 arg1 waves 1632:1636 spiral waves 1625:1636 spiral waves 1625:1636 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 0 85 theme dynamics 47:54 arg1 modeling 10:17 In-silico modeling 0:17 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 0:107 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 9 86 theme in-silico 1286:1294 arg1 models 1296:1301 the in-silico models 1282:1301 the in-silico models 1282:1301 From the in-silico models, reduced glycosylation was shown to shorten the repolarization phase of cardiac APs, thereby influencing electrical propagation in cardiac fibers and tissues. 24403418 0 87 theme hERG 59:62 arg1 channels 68:75 hERG ion channels 59:75 hERG ion channels 59:75 In-silico modeling of glycosylation modulation dynamics in hERG ion channels and cardiac electrical signals. 24403418 4 88 theme reduced 499:505 arg1 glycosylation 507:519 reduced glycosylation 499:519 reduced glycosylation 499:519 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 10 89 link derived 1491:1497 arg1 electrocardiogram 1499:1515 derived electrocardiogram 1491:1515 derived electrocardiogram 1491:1515 In addition, the patterns of derived electrocardiogram show that reduced glycosylation of hERG channel shortens the QT interval and decreases the re-entry rate of spiral waves. 24403418 2 90 theme cardiac 326:332 arg1 rhythms 334:340 abnormal cardiac rhythms 317:340 abnormal cardiac rhythms 317:340 A slight change in ion channel activity may affect the AP waveform, thereby potentially increasing susceptibility to abnormal cardiac rhythms. 24403418 4 91 theme related 569:575 arg1 channel 583:589 hERG (human ether-a-go-go related gene) channel 543:589 hERG (human ether-a-go-go related gene) channel 543:589 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 4 91 theme related 569:575 arg1 gene 577:580 human ether-a-go-go related gene 549:580 human ether-a-go-go related gene 549:580 However, little is known about how reduced glycosylation impacts the gating of hERG (human ether-a-go-go related gene) channel, which is partially responsible for late phase 2 and phase 3 of the AP. 24403418 11 92 theme new 1658:1660 arg1 targets 1677:1683 new pharmaceutical targets 1658:1683 new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders 1658:1748 This work suggests new pharmaceutical targets for the long QT syndrome and potentially other cardiac disorders. 24403418 3 93 theme glycan 445:450 arg1 structures 452:461 glycan structures 445:461 glycan structures 445:461 Cardiac ion channels are heavily glycosylated, with up to 30% of a mature protein's mass comprised of glycan structures. 24403418 8 94 dep cells 1248:1252 arg1 i.e. 1234:1237 i.e. 1234:1237 i.e. 1234:1237 Further, we developed in-silico models to simulate glycosylation-channel interactions and predict the effects of reduced glycosylation on multiscale cardiac processes (i.e., cardiac cells, 1-D and 2-D tissues). 27697163 7 0 theme mannose/galactose 1311:1327 arg1 ratio 1329:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 2 1 theme synthetic 628:636 arg1 juice 638:642 synthetic juice 628:642 synthetic juice 628:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 1 2 theme wall 289:292 arg1 origin 294:299 cell wall origin 284:299 cell wall origin 284:299 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 9 3 theme yeast 1756:1760 arg1 species 1762:1768 both yeast species 1751:1768 both yeast species 1751:1768 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 10 4 theme possible 1859:1866 arg1 role 1868:1871 a possible role 1857:1871 a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint 1857:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 10 4 theme possible 1859:1866 arg1 tool 1956:1959 a taxonomic tool 1944:1959 a taxonomic tool for this genus 1944:1974 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 10 5 theme MALDI-TOF 1882:1890 arg1 screening 1892:1900 rapid MALDI-TOF screening 1876:1900 rapid MALDI-TOF screening of N-glycans compositional fingerprint 1876:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 1 6 theme Schizosaccharomyces 212:230 arg1 release 238:244 the Schizosaccharomyces genus release 208:244 the Schizosaccharomyces genus release 208:244 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 2 7 theme same 596:599 arg1 conditions 614:623 the same fermentative conditions 592:623 the same fermentative conditions of synthetic juice 592:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 6 8 theme released 1193:1200 arg1 polysaccharides 1202:1216 the released polysaccharides 1189:1216 the released polysaccharides 1189:1216 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 5 9 with strains 972:978 arg1 patterns 1018:1025 different malic acid consumption patterns 985:1025 different malic acid consumption patterns along with high polysaccharide release 985:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 11 10 theme exogenous 2161:2169 arg1 preparations 2186:2197 exogenous polysaccharide preparations 2161:2197 exogenous polysaccharide preparations 2161:2197 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 7 11 located observed 1378:1385 arg2 percentage 1351:1360 a different percentage 1339:1360 a different percentage of proteins 1339:1372 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 7 11 located observed 1378:1385 arg2 ratio 1329:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 7 11 located observed 1378:1385 arg1 polysaccharides 1394:1408 the polysaccharides 1390:1408 the polysaccharides released by S. pombe as compared to S. japonicus 1390:1457 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 10 12 theme compositional 1915:1927 arg1 fingerprint 1929:1939 N-glycans compositional fingerprint 1905:1939 N-glycans compositional fingerprint 1905:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 1 13 theme high 248:251 arg1 quantity 253:260 a high quantity 246:260 a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation 246:353 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 4 14 theme color 931:935 arg1 stability 937:945 color stability 931:945 color stability of wine 931:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 8 15 theme glycoprotein 1534:1545 arg1 presence 1520:1527 the presence 1516:1527 the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus 1516:1618 Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. 27697163 6 16 dep species 1264:1270 arg1 S. pombe 1272:1279 S. pombe 1272:1279 S. pombe 1272:1279 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 6 16 dep species 1264:1270 arg1 species 1264:1270 the two yeast species S. pombe and S. japonicus 1250:1296 the two yeast species S. pombe and S. japonicus 1250:1296 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 6 16 dep species 1264:1270 arg1 S. japonicus 1285:1296 S. japonicus 1285:1296 S. japonicus 1285:1296 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 5 17 theme modified 1103:1110 arg1 wines 1112:1116 naturally modified wines 1093:1116 naturally modified wines 1093:1116 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 10 18 theme taxonomic 1946:1954 arg1 role 1868:1871 a possible role 1857:1871 a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint 1857:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 10 18 theme taxonomic 1946:1954 arg1 tool 1956:1959 a taxonomic tool 1944:1959 a taxonomic tool for this genus 1944:1974 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 1 19 theme present 157:163 arg1 work 165:168 The present work 153:168 The present work 153:168 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 7 20 theme different 1301:1309 arg1 ratio 1329:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 2 21 theme juice 638:642 arg1 conditions 614:623 the same fermentative conditions 592:623 the same fermentative conditions of synthetic juice 592:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 2 22 theme yeast 573:577 arg1 strain 579:584 a commercial Saccharomyces cerevisiae yeast strain 535:584 a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice 535:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 11 23 from release 1993:1999 arg1 media 2008:2012 the media 2004:2012 the media 2004:2012 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 11 23 from release 1993:1999 arg1 galactomannoproteins 2029:2048 particular galactomannoproteins 2018:2048 particular galactomannoproteins in significant amounts 2018:2071 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 0 24 dep quantification 116:129 arg1 polysaccharides 10:24 Cell wall polysaccharides 0:24 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus 0:113 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus: quantification and characterization. 27697163 2 25 theme Saccharomyces 548:560 arg1 strain 579:584 a commercial Saccharomyces cerevisiae yeast strain 535:584 a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice 535:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 9 26 theme carbohydrate 1651:1662 arg1 moieties 1664:1671 carbohydrate moieties 1651:1671 carbohydrate moieties 1651:1671 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 0 27 theme Cell 0:3 arg1 polysaccharides 10:24 Cell wall polysaccharides 0:24 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus 0:113 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus: quantification and characterization. 27697163 2 28 dep polysaccharides 466:480 arg1 times 500:504 times 500:504 times 500:504 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 5 29 theme strains 972:978 arg1 presence 960:967 The presence 956:967 The presence of strains with different malic acid consumption patterns along with high polysaccharide release 956:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 2 30 dep strains 435:441 arg1 Schizosaccharomyces 409:427 the Schizosaccharomyces yeast strains 405:441 the Schizosaccharomyces yeast strains 405:441 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 11 31 theme significant 2053:2063 arg1 amounts 2065:2071 significant amounts 2053:2071 significant amounts 2053:2071 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 5 32 theme different 985:993 arg1 patterns 1018:1025 different malic acid consumption patterns 985:1025 different malic acid consumption patterns along with high polysaccharide release 985:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 4 33 theme acid 872:875 arg1 levels 854:859 high levels 849:859 high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine 849:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 0 34 theme alcoholic 46:54 arg1 fermentation 56:67 the alcoholic fermentation 42:67 the alcoholic fermentation 42:67 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus: quantification and characterization. 27697163 1 35 theme fermentation 342:353 arg1 onset 319:323 the onset 315:323 the onset of the alcoholic fermentation 315:353 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 7 36 dep S. pombe 1422:1429 arg1 compared 1434:1441 compared 1434:1441 compared to S. japonicus 1434:1457 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 7 37 theme proteins 1365:1372 arg1 percentage 1351:1360 a different percentage 1339:1360 a different percentage of proteins 1339:1372 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 7 37 theme proteins 1365:1372 arg1 ratio 1329:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio 1299:1333 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 4 38 theme high 849:852 arg1 levels 854:859 high levels 849:859 high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine 849:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 4 39 theme pyruvic 864:870 arg1 acid 872:875 pyruvic acid 864:875 pyruvic acid 864:875 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 4 39 theme pyruvic 864:870 arg1 compound 918:925 an important compound 905:925 an important compound for color stability of wine 905:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 11 40 theme Polysaccharides 1977:1991 arg1 release 1993:1999 Polysaccharides release 1977:1999 Polysaccharides release 1977:1999 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 1 41 theme polysaccharides 265:279 arg1 quantity 253:260 a high quantity 246:260 a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation 246:353 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 5 42 theme polysaccharide 1043:1056 arg1 release 1058:1064 high polysaccharide release 1038:1064 high polysaccharide release 1038:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 2 43 theme alcoholic 374:382 arg1 fermentation 384:395 the alcoholic fermentation 370:395 the alcoholic fermentation 370:395 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 1 44 theme cell 284:287 arg1 origin 294:299 cell wall origin 284:299 cell wall origin 284:299 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 11 45 theme industrial 2136:2145 arg1 production 2147:2156 the industrial production 2132:2156 the industrial production of exogenous polysaccharide preparations 2132:2197 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 1 46 theme origin 294:299 arg1 polysaccharides 265:279 polysaccharides 265:279 polysaccharides of cell wall origin 265:299 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 10 47 theme screening 1892:1900 arg1 role 1868:1871 a possible role 1857:1871 a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint 1857:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 10 47 theme screening 1892:1900 arg1 tool 1956:1959 a taxonomic tool 1944:1959 a taxonomic tool for this genus 1944:1974 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 4 48 theme important 908:916 arg1 acid 872:875 pyruvic acid 864:875 pyruvic acid 864:875 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 4 48 theme important 908:916 arg1 compound 918:925 an important compound 905:925 an important compound for color stability of wine 905:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 3 49 located found 684:688 arg1 media 693:697 media 693:697 media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe 693:790 A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe. 27697163 3 49 located found 684:688 arg2 content 654:660 A higher content 645:660 A higher content of polysaccharide 645:678 A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe. 27697163 11 50 theme polysaccharide 2171:2184 arg1 preparations 2186:2197 exogenous polysaccharide preparations 2161:2197 exogenous polysaccharide preparations 2161:2197 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 8 51 gly glycoprotein 1534:1545 arg1 glycoprotein 1534:1545 a glycoprotein 1532:1545 a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus 1532:1618 Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. 27697163 6 52 theme polysaccharides 1202:1216 arg1 analysis 1177:1184 The chemical analysis 1164:1184 The chemical analysis of the released polysaccharides 1164:1216 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 10 53 theme rapid 1876:1880 arg1 screening 1892:1900 rapid MALDI-TOF screening 1876:1900 rapid MALDI-TOF screening of N-glycans compositional fingerprint 1876:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 1 54 theme genus 232:236 arg1 release 238:244 the Schizosaccharomyces genus release 208:244 the Schizosaccharomyces genus release 208:244 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 11 55 theme preparations 2186:2197 arg1 production 2147:2156 the industrial production 2132:2156 the industrial production of exogenous polysaccharide preparations 2132:2197 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 4 56 theme wine 950:953 arg1 stability 937:945 color stability 931:945 color stability of wine 931:953 Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. 27697163 9 57 theme Mass 1621:1624 arg1 analysis 1639:1646 Mass spectrometry analysis 1621:1646 Mass spectrometry analysis of carbohydrate moieties 1621:1671 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 5 58 theme enhanced 1123:1130 arg1 mouth 1132:1136 mouth feel 1132:1141 mouth feel 1132:1141 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 10 59 theme fingerprint 1929:1939 arg1 screening 1892:1900 rapid MALDI-TOF screening 1876:1900 rapid MALDI-TOF screening of N-glycans compositional fingerprint 1876:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 1 60 dep quantity 253:260 arg1 yeasts 188:193 yeasts 188:193 yeasts belonging to the Schizosaccharomyces genus release 188:244 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 11 61 from galactomannoproteins 2029:2048 arg1 amounts 2065:2071 significant amounts 2053:2071 significant amounts 2053:2071 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 10 62 theme N-glycans 1905:1913 arg1 fingerprint 1929:1939 N-glycans compositional fingerprint 1905:1939 N-glycans compositional fingerprint 1905:1939 These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. 27697163 8 63 theme molecular 1554:1562 arg1 size 1564:1567 a molecular size 1552:1567 a molecular size around 32-33 kDa only for the species S. japonicus 1552:1618 Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. 27697163 3 64 theme polysaccharide 665:678 arg1 content 654:660 A higher content 645:660 A higher content of polysaccharide 645:678 A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe. 27697163 2 65 theme fermentative 601:612 arg1 conditions 614:623 the same fermentative conditions 592:623 the same fermentative conditions of synthetic juice 592:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 9 66 theme spectrometry 1626:1637 arg1 analysis 1639:1646 Mass spectrometry analysis 1621:1646 Mass spectrometry analysis of carbohydrate moieties 1621:1671 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 7 67 theme different 1341:1349 arg1 percentage 1351:1360 a different percentage 1339:1360 a different percentage of proteins 1339:1372 A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. 27697163 5 68 theme wines 1112:1116 arg1 production 1079:1088 production 1079:1088 production of naturally modified wines 1079:1116 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 6 69 theme yeast 1258:1262 arg1 S. pombe 1272:1279 S. pombe 1272:1279 S. pombe 1272:1279 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 6 69 theme yeast 1258:1262 arg1 species 1264:1270 the two yeast species S. pombe and S. japonicus 1250:1296 the two yeast species S. pombe and S. japonicus 1250:1296 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 6 69 theme yeast 1258:1262 arg1 S. japonicus 1285:1296 S. japonicus 1285:1296 S. japonicus 1285:1296 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 2 70 dep times 500:504 arg1 higher 506:511 higher 506:511 higher 506:511 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 5 71 theme malic 995:999 arg1 patterns 1018:1025 different malic acid consumption patterns 985:1025 different malic acid consumption patterns along with high polysaccharide release 985:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 5 72 theme acid 1001:1004 arg1 patterns 1018:1025 different malic acid consumption patterns 985:1025 different malic acid consumption patterns along with high polysaccharide release 985:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 5 73 theme reduced 1147:1153 arg1 acidity 1155:1161 reduced acidity 1147:1161 reduced acidity 1147:1161 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 5 74 theme consumption 1006:1016 arg1 patterns 1018:1025 different malic acid consumption patterns 985:1025 different malic acid consumption patterns along with high polysaccharide release 985:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 2 75 theme commercial 537:546 arg1 strain 579:584 a commercial Saccharomyces cerevisiae yeast strain 535:584 a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice 535:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 11 76 theme particular 2018:2027 arg1 galactomannoproteins 2029:2048 particular galactomannoproteins 2018:2048 particular galactomannoproteins in significant amounts 2018:2071 Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. 27697163 0 77 theme wall 5:8 arg1 polysaccharides 10:24 Cell wall polysaccharides 0:24 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus 0:113 Cell wall polysaccharides released during the alcoholic fermentation by Schizosaccharomyces pombe and S. japonicus: quantification and characterization. 27697163 9 78 theme moieties 1664:1671 arg1 analysis 1639:1646 Mass spectrometry analysis 1621:1646 Mass spectrometry analysis of carbohydrate moieties 1621:1671 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 2 79 theme polysaccharides 466:480 arg1 quantity 454:461 a quantity 452:461 a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice 452:642 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 2 80 dep Saccharomyces 548:560 arg1 cerevisiae 562:571 cerevisiae 562:571 cerevisiae 562:571 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 9 81 theme similar 1680:1686 arg1 proportions 1688:1698 similar proportions 1680:1698 similar proportions 1680:1698 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 1 82 dep demonstrates 170:181 arg1 quantity 253:260 a high quantity 246:260 a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation 246:353 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 8 83 theme proteins 1476:1483 arg1 Analysis 1460:1467 Analysis 1460:1467 Analysis of the proteins released in the media 1460:1505 Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. 27697163 2 84 theme yeast 429:433 arg1 strains 435:441 yeast strains 429:441 the Schizosaccharomyces yeast strains 405:441 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 1 85 theme alcoholic 332:340 arg1 fermentation 342:353 the alcoholic fermentation 328:353 the alcoholic fermentation 328:353 The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. 27697163 9 86 theme N-glycan 1710:1717 arg1 chains 1719:1724 the N-glycan chains 1706:1724 the N-glycan chains released in the media by both yeast species 1706:1768 Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. 27697163 2 87 theme 3-7 496:498 arg1 times 500:504 times 500:504 times 500:504 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27697163 8 88 with glycoprotein 1534:1545 arg1 size 1564:1567 a molecular size 1552:1567 a molecular size around 32-33 kDa only for the species S. japonicus 1552:1618 Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. 27697163 3 89 theme higher 647:652 arg1 content 654:660 A higher content 645:660 A higher content of polysaccharide 645:678 A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe. 27697163 6 90 theme chemical 1168:1175 arg1 analysis 1177:1184 The chemical analysis 1164:1184 The chemical analysis of the released polysaccharides 1164:1216 The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. 27697163 5 91 dep mouth 1132:1136 arg1 feel 1138:1141 feel 1138:1141 feel 1138:1141 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 5 92 theme high 1038:1041 arg1 release 1058:1064 high polysaccharide release 1038:1064 high polysaccharide release 1038:1064 The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. 27697163 2 93 theme fermentation 384:395 arg1 end 363:365 the end 359:365 the end of the alcoholic fermentation 359:395 By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. 27073020 4 0 theme glycan 854:859 arg1 changes 861:867 different glycan changes 844:867 different glycan changes 844:867 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 3 1 gly glycoproteins 502:514 arg1 glycoproteins 502:514 plasma glycoproteins 495:514 plasma glycoproteins 495:514 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 4 2 theme different 719:727 arg1 concentrations 729:742 Two different concentrations 715:742 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) 715:780 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 3 3 from blood 637:641 arg1 detectable 623:632 detectable 623:632 detectable 623:632 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 3 3 from blood 637:641 arg1 model 575:579 a good model 568:579 a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting 568:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 4 4 theme different 844:852 arg1 changes 861:867 different glycan changes 844:867 different glycan changes 844:867 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 7 5 theme tetra-antennary 1260:1274 arg1 structures 1276:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 7 5 theme tetra-antennary 1260:1274 arg1 result 1326:1331 a result 1324:1331 a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1324:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 1 6 theme few 241:243 arg1 studies 245:251 few studies 241:251 few studies 241:251 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 3 7 from changes 599:605 arg1 HCC 610:612 HCC 610:612 HCC 610:612 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 4 8 theme global 806:811 arg1 methylation 820:830 global genome methylation 806:830 global genome methylation 806:830 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 5 9 theme 5-aza-2dC 954:962 arg1 treatment 964:972 the 5-aza-2dC treatment 950:972 the 5-aza-2dC treatment 950:972 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 3 10 theme analytic 674:681 arg1 which 644:648 which 644:648 which 644:648 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 3 10 theme analytic 674:681 arg1 material 683:690 an easily accessible analytic material 653:690 an easily accessible analytic material in a clinical setting 653:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 6 11 theme glycan 1179:1184 arg1 changes 1186:1192 the glycan changes 1175:1192 the glycan changes consistently induced by both doses of 5-aza-2dC 1175:1240 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 2 12 attach derived 428:434 arg2 line 423:426 the HepG2 cell line 408:426 the HepG2 cell line derived from hepatocellular carcinoma (HCC) 408:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 2 12 attach derived 428:434 arg1 HCC 467:469 HCC 467:469 HCC 467:469 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 2 12 attach derived 428:434 arg1 carcinoma 456:464 hepatocellular carcinoma 441:464 hepatocellular carcinoma (HCC) 441:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 6 13 theme HepG2 1093:1097 arg1 profile 1113:1119 the HepG2 glycosylation profile 1089:1119 the HepG2 glycosylation profile 1089:1119 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 6 14 theme MGAT3 1139:1143 arg1 gene 1145:1148 the MGAT3 gene 1135:1148 the MGAT3 gene 1135:1148 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 6 14 theme MGAT3 1139:1143 arg1 responsible 1159:1169 responsible 1159:1169 responsible 1159:1169 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 7 15 gly Core-fucosylated 1243:1258 arg1 structures 1276:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 7 15 gly Core-fucosylated 1243:1258 arg1 result 1326:1331 a result 1324:1331 a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1324:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 1 16 from Changes 143:149 arg1 N-glycosylation 154:168 N-glycosylation 154:168 N-glycosylation of plasma proteins 154:187 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 3 17 theme plasma 495:500 arg1 glycoproteins 502:514 plasma glycoproteins 495:514 plasma glycoproteins 495:514 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 6 18 theme glyco-gene 1063:1072 arg1 expression 1074:1083 glyco-gene expression 1063:1083 glyco-gene expression 1063:1083 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 7 19 theme gene 1357:1360 arg1 promoter 1362:1369 hypomethylated MGAT3 gene promoter 1336:1369 hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1336:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 4 20 theme genome 813:818 arg1 methylation 820:830 global genome methylation 806:830 global genome methylation 806:830 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 2 21 from impact 345:350 arg1 N-glycome 378:386 the N-glycome 374:386 the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC) 374:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 2 22 theme cell 418:421 arg1 line 423:426 the HepG2 cell line 408:426 the HepG2 cell line derived from hepatocellular carcinoma (HCC) 408:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 1 23 theme plasma 173:178 arg1 proteins 180:187 plasma proteins 173:187 plasma proteins 173:187 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 2 24 theme line 423:426 arg1 secretome 395:403 the secretome 391:403 the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC) 391:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 3 25 theme accessible 663:672 arg1 which 644:648 which 644:648 which 644:648 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 3 25 theme accessible 663:672 arg1 material 683:690 an easily accessible analytic material 653:690 an easily accessible analytic material in a clinical setting 653:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 5 26 theme global 989:994 arg1 hypomethylation 1003:1017 global genome hypomethylation 989:1017 global genome hypomethylation 989:1017 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 5 27 theme expression 927:936 arg1 result 979:984 a result 977:984 a result of global genome hypomethylation 977:1017 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 5 27 theme expression 927:936 arg1 level 938:942 expression level 927:942 expression level 927:942 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 1 28 theme proteins 180:187 arg1 N-glycosylation 154:168 N-glycosylation 154:168 N-glycosylation of plasma proteins 154:187 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 0 29 theme DNA 0:2 arg1 hypomethylation 4:18 DNA hypomethylation 0:18 DNA hypomethylation 0:18 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 3 30 theme clinical 697:704 arg1 setting 706:712 a clinical setting 695:712 a clinical setting 695:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 6 31 from changes 1052:1058 arg1 expression 1074:1083 glyco-gene expression 1063:1083 glyco-gene expression 1063:1083 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 7 32 theme increased 1383:1391 arg1 expression 1393:1402 increased expression 1383:1402 increased expression of this gene 1383:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 2 33 theme methylation 359:369 arg1 impact 345:350 the impact 341:350 the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC) 341:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 2 34 theme HepG2 412:416 arg1 line 423:426 the HepG2 cell line 408:426 the HepG2 cell line derived from hepatocellular carcinoma (HCC) 408:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 1 35 theme exact 265:269 arg1 mechanism 271:279 the exact mechanism 261:279 the exact mechanism involved in aberrant protein glycosylation 261:322 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 7 36 theme gene 1412:1415 arg1 expression 1393:1402 increased expression 1383:1402 increased expression of this gene 1383:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 3 37 from material 683:690 arg1 setting 706:712 a clinical setting 695:712 a clinical setting 695:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 7 38 theme hypomethylated 1336:1349 arg1 promoter 1362:1369 hypomethylated MGAT3 gene promoter 1336:1369 hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1336:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 0 39 theme glycoproteins 128:140 arg1 N-glycosylation 100:114 N-glycosylation 100:114 N-glycosylation of secreted glycoproteins 100:140 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 5 40 theme hypomethylation 1003:1017 arg1 result 979:984 a result 977:984 a result of global genome hypomethylation 977:1017 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 5 40 theme hypomethylation 1003:1017 arg1 level 938:942 expression level 927:942 expression level 927:942 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 7 41 theme MGAT3 1351:1355 arg1 promoter 1362:1369 hypomethylated MGAT3 gene promoter 1336:1369 hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1336:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 1 42 theme many 205:208 arg1 types 210:214 many types 205:214 many types of cancer 205:224 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 2 43 theme hepatocellular 441:454 arg1 HCC 467:469 HCC 467:469 HCC 467:469 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 2 43 theme hepatocellular 441:454 arg1 carcinoma 456:464 hepatocellular carcinoma 441:464 hepatocellular carcinoma (HCC) 441:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 0 44 theme secreted 119:126 arg1 glycoproteins 128:140 secreted glycoproteins 119:140 secreted glycoproteins 119:140 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 6 45 theme 5-aza-2dC 1232:1240 arg1 doses 1223:1227 both doses 1218:1227 both doses of 5-aza-2dC 1218:1240 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 0 46 theme gene 56:59 arg1 expression 32:41 expression 32:41 expression of the MGAT3 gene in HepG2 cells 32:74 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 1 47 theme aberrant 293:300 arg1 glycosylation 310:322 aberrant protein glycosylation 293:322 aberrant protein glycosylation 293:322 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 3 48 theme HepG2 546:550 arg1 cells 552:556 the HepG2 cells 542:556 the HepG2 cells 542:556 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 0 49 gly N-glycosylation 100:114 arg1 glycoproteins 128:140 secreted glycoproteins 119:140 secreted glycoproteins 119:140 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 0 50 theme MGAT3 50:54 arg1 gene 56:59 the MGAT3 gene 46:59 the MGAT3 gene 46:59 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 1 51 theme protein 302:308 arg1 glycosylation 310:322 aberrant protein glycosylation 293:322 aberrant protein glycosylation 293:322 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 1 52 located observed 193:200 arg2 Changes 143:149 Changes 143:149 Changes in N-glycosylation of plasma proteins 143:187 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 1 52 located observed 193:200 arg1 types 210:214 many types 205:214 many types of cancer 205:224 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 2 53 from N-glycome 378:386 arg1 secretome 395:403 the secretome 391:403 the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC) 391:470 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 5 54 theme glyco-genes 898:908 arg1 percent 884:890 Around twenty percent 870:890 Around twenty percent of 84 glyco-genes analysed 870:917 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 1 55 gly N-glycosylation 154:168 arg1 proteins 180:187 plasma proteins 173:187 plasma proteins 173:187 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 0 56 from changes 89:95 arg1 N-glycosylation 100:114 N-glycosylation 100:114 N-glycosylation of secreted glycoproteins 100:140 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 4 57 theme 5-aza-2'-deoxycytidine 747:768 arg1 concentrations 729:742 Two different concentrations 715:742 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) 715:780 Two different concentrations of 5-aza-2'-deoxycytidine (5-aza-2dC) differentially affected global genome methylation and induced different glycan changes. 27073020 6 58 theme correlation 1022:1032 arg1 study 1034:1038 A correlation study 1020:1038 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile 1020:1119 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 3 59 theme glycoproteins 502:514 arg1 majority 483:490 the majority 479:490 the majority of plasma glycoproteins 479:514 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 3 60 theme glycosylation 585:597 arg1 changes 599:605 glycosylation changes 585:605 glycosylation changes in HCC 585:612 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 0 61 gly glycoproteins 128:140 arg1 glycoproteins 128:140 secreted glycoproteins 119:140 secreted glycoproteins 119:140 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 1 62 theme cancer 219:224 arg1 types 210:214 many types 205:214 many types of cancer 205:224 Changes in N-glycosylation of plasma proteins are observed in many types of cancer, nevertheless, few studies suggest the exact mechanism involved in aberrant protein glycosylation. 27073020 0 63 theme HepG2 64:68 arg1 cells 70:74 HepG2 cells 64:74 HepG2 cells 64:74 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 3 64 theme good 570:573 arg1 model 575:579 a good model 568:579 a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting 568:712 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 3 64 theme good 570:573 arg1 detectable 623:632 detectable 623:632 detectable 623:632 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 2 65 theme DNA 355:357 arg1 methylation 359:369 DNA methylation 355:369 DNA methylation 355:369 Here we studied the impact of DNA methylation on the N-glycome in the secretome of the HepG2 cell line derived from hepatocellular carcinoma (HCC). 27073020 7 66 theme promoter 1362:1369 arg1 result 1326:1331 a result 1324:1331 a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1324:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 7 66 theme promoter 1362:1369 arg1 structures 1276:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 3 67 from detectable 623:632 arg1 blood 637:641 blood 637:641 blood 637:641 Since the majority of plasma glycoproteins originate from the liver, the HepG2 cells represent a good model for glycosylation changes in HCC that are detectable in blood, which is an easily accessible analytic material in a clinical setting. 27073020 0 68 from expression 32:41 arg1 cells 70:74 HepG2 cells 64:74 HepG2 cells 64:74 DNA hypomethylation upregulates expression of the MGAT3 gene in HepG2 cells and leads to changes in N-glycosylation of secreted glycoproteins. 27073020 5 69 theme genome 996:1001 arg1 hypomethylation 1003:1017 global genome hypomethylation 989:1017 global genome hypomethylation 989:1017 Around twenty percent of 84 glyco-genes analysed changed expression level after the 5-aza-2dC treatment as a result of global genome hypomethylation. 27073020 6 70 from profile 1113:1119 arg1 expression 1074:1083 glyco-gene expression 1063:1083 glyco-gene expression 1063:1083 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 6 71 theme glycosylation 1099:1111 arg1 profile 1113:1119 the HepG2 glycosylation profile 1089:1119 the HepG2 glycosylation profile 1089:1119 A correlation study between the changes in glyco-gene expression and the HepG2 glycosylation profile suggests that the MGAT3 gene might be responsible for the glycan changes consistently induced by both doses of 5-aza-2dC. 27073020 7 72 theme Core-fucosylated 1243:1258 arg1 structures 1276:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures 1243:1285 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27073020 7 72 theme Core-fucosylated 1243:1258 arg1 result 1326:1331 a result 1324:1331 a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene 1324:1415 Core-fucosylated tetra-antennary structures were decreased in quantity likely as a result of hypomethylated MGAT3 gene promoter followed by increased expression of this gene. 27693835 0 0 theme citrus 93:98 arg1 peels 100:104 citrus peels 93:104 citrus peels (Citrus unshiu) 93:120 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 0 0 theme citrus 93:98 arg1 unshiu 114:119 Citrus unshiu 107:119 Citrus unshiu 107:119 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 6 1 theme AceA-containing 861:875 arg1 nonasaccharide 877:890 an AceA-containing nonasaccharide 858:890 an AceA-containing nonasaccharide 858:890 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 5 2 theme methylation 630:640 arg1 analysis 642:649 methylation analysis 630:649 methylation analysis 630:649 The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. 27693835 1 3 theme study 139:143 arg1 aim 127:129 The aim 123:129 The aim of this study 123:143 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 3 4 theme in 309:310 arg1 metastasis 322:331 in vivo lung metastasis 309:331 in vivo lung metastasis of Colon26-M3.1 309:347 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 4 5 theme glycosyl 497:504 arg1 linkages 506:513 22 different glycosyl linkages 484:513 22 different glycosyl linkages 484:513 CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). 27693835 3 6 theme Colon26-M3.1 336:347 arg1 metastasis 322:331 in vivo lung metastasis 309:331 in vivo lung metastasis of Colon26-M3.1 309:347 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 4 7 theme different 487:495 arg1 linkages 506:513 22 different glycosyl linkages 484:513 22 different glycosyl linkages 484:513 CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). 27693835 0 8 theme peels 100:104 arg1 digest 83:88 the pectinase digest 69:88 the pectinase digest of citrus peels (Citrus unshiu) 69:120 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 5 9 theme oligosaccharide 652:666 arg1 analysis 668:675 oligosaccharide analysis 652:675 oligosaccharide analysis 652:675 The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. 27693835 2 10 theme pectinase 265:273 arg1 digestion 275:283 the pectinase digestion 261:283 the pectinase digestion of citrus peels 261:299 CPE-II was purified by the pectinase digestion of citrus peels. 27693835 8 11 theme anti-metastatic 1141:1155 arg1 CPE-II 1157:1162 the anti-metastatic CPE-II 1137:1162 the anti-metastatic CPE-II 1137:1162 Thus, we propose that the anti-metastatic CPE-II is primarily present as an RG-II dimer. 27693835 8 11 theme anti-metastatic 1141:1155 arg1 present 1177:1183 present 1177:1183 present 1177:1183 Thus, we propose that the anti-metastatic CPE-II is primarily present as an RG-II dimer. 27693835 0 12 theme Citrus 107:112 arg1 peels 100:104 citrus peels 93:104 citrus peels (Citrus unshiu) 93:120 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 0 12 theme Citrus 107:112 arg1 unshiu 114:119 Citrus unshiu 107:119 Citrus unshiu 107:119 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 6 13 theme 1→5 848:850 arg1 -Dha 852:855 Araf-(1→5)-Dha 842:855 Araf-(1→5)-Dha 842:855 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 5 14 theme sugar 611:615 arg1 composition 617:627 sugar composition 611:627 sugar composition 611:627 The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. 27693835 6 15 theme acid 778:781 arg1 hydrolysis 783:792 partial acid hydrolysis 770:792 partial acid hydrolysis 770:792 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 3 16 theme 10μg 368:371 arg1 administration 350:363 administration 350:363 administration of 10μg of CPE-II per mouse 350:391 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 4 17 theme different 454:462 arg1 monosaccharides 464:478 15 different monosaccharides 451:478 15 different monosaccharides 451:478 CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). 27693835 6 18 theme partial 770:776 arg1 hydrolysis 783:792 partial acid hydrolysis 770:792 partial acid hydrolysis 770:792 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 3 19 theme 81.3 400:403 arg1 % 404:404 % 404:404 % 404:404 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 0 20 theme Structural 0:9 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 0:121 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 2 21 theme peels 295:299 arg1 digestion 275:283 the pectinase digestion 261:283 the pectinase digestion of citrus peels 261:299 CPE-II was purified by the pectinase digestion of citrus peels. 27693835 3 22 theme % 404:404 arg1 inhibition 406:415 81.3% inhibition 400:415 81.3% inhibition of metastasis 400:429 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 0 23 theme anti-metastatic 26:40 arg1 rhamnogalacturonan 42:59 anti-metastatic rhamnogalacturonan II 26:62 anti-metastatic rhamnogalacturonan II 26:62 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 4 24 theme rhamnogalacturonan 534:551 arg1 characteristic 516:529 characteristic 516:529 characteristic 516:529 CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). 27693835 7 25 theme CPE-II 1023:1028 arg1 weight 1013:1018 The molecular weight 999:1018 The molecular weight of CPE-II 999:1028 The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. 27693835 5 26 theme primary 569:575 arg1 structure 577:585 The primary structure 565:585 The primary structure 565:585 The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. 27693835 2 27 theme citrus 288:293 arg1 peels 295:299 citrus peels 288:299 citrus peels 288:299 CPE-II was purified by the pectinase digestion of citrus peels. 27693835 7 28 dep 5kDa 1065:1068 arg1 to 1062:1063 to 1062:1063 to 1062:1063 The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. 27693835 1 29 with peels 198:202 arg1 property 228:235 an anti-metastatic property 209:235 an anti-metastatic property 209:235 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 6 30 theme Araf- 842:846 arg1 -Dha 852:855 Araf-(1→5)-Dha 842:855 Araf-(1→5)-Dha 842:855 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 31 theme acid-rich 907:915 arg1 oligosaccharide 917:931 an uronic acid-rich oligosaccharide 897:931 an uronic acid-rich oligosaccharide 897:931 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 3 32 theme metastasis 420:429 arg1 inhibition 406:415 81.3% inhibition 400:415 81.3% inhibition of metastasis 400:429 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 1 33 theme citrus 191:196 arg1 peels 198:202 citrus peels 191:202 citrus peels with an anti-metastatic property 191:235 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 0 34 theme rhamnogalacturonan 42:59 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 0:121 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 6 35 theme uronic 900:905 arg1 oligosaccharide 917:931 an uronic acid-rich oligosaccharide 897:931 an uronic acid-rich oligosaccharide 897:931 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 0 36 from digest 83:88 arg1 elucidation 11:21 Structural elucidation 0:21 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 0:121 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 7 37 theme <2.0 1087:1090 arg1 value 1078:1082 a pH value 1073:1082 a pH value of <2.0 1073:1090 The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. 27693835 3 38 dep in 309:310 arg1 vivo 312:315 vivo 312:315 vivo 312:315 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 3 39 theme CPE-II 376:381 arg1 administration 350:363 administration 350:363 administration of 10μg of CPE-II per mouse 350:391 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 27693835 0 40 theme pectinase 73:81 arg1 digest 83:88 the pectinase digest 69:88 the pectinase digest of citrus peels (Citrus unshiu) 69:120 Structural elucidation of anti-metastatic rhamnogalacturonan II from the pectinase digest of citrus peels (Citrus unshiu). 27693835 7 41 theme pH 1075:1076 arg1 value 1078:1082 a pH value 1073:1082 a pH value of <2.0 1073:1090 The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. 27693835 7 42 theme molecular 1003:1011 arg1 weight 1013:1018 The molecular weight 999:1018 The molecular weight of CPE-II 999:1028 The molecular weight of CPE-II was observed to decrease from 9 to 5kDa at a pH value of <2.0, as observed by HPSEC. 27693835 6 43 theme 1→5 832:834 arg1 -Kdo 836:839 Rhap-(1→5)-Kdo 826:839 Rhap-(1→5)-Kdo 826:839 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 8 44 theme RG-II 1191:1195 arg1 dimer 1197:1201 an RG-II dimer 1188:1201 an RG-II dimer 1188:1201 Thus, we propose that the anti-metastatic CPE-II is primarily present as an RG-II dimer. 27693835 4 45 theme characteristic 516:529 arg1 monosaccharides 464:478 15 different monosaccharides 451:478 15 different monosaccharides 451:478 CPE-II consists of 15 different monosaccharides and 22 different glycosyl linkages, characteristic of rhamnogalacturonan II (RG-II). 27693835 1 46 located found 182:186 arg2 polysaccharide 167:180 a polysaccharide 165:180 a polysaccharide found in citrus peels with an anti-metastatic property 165:235 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 1 46 located found 182:186 arg1 peels 198:202 citrus peels 191:202 citrus peels with an anti-metastatic property 191:235 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 6 47 theme main 987:990 arg1 chain 992:996 an α-(1→4)-galacturono-oligosaccharide main chain 948:996 an α-(1→4)-galacturono-oligosaccharide main chain 948:996 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 48 theme Rhap- 826:830 arg1 -Kdo 836:839 Rhap-(1→5)-Kdo 826:839 Rhap-(1→5)-Kdo 826:839 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 49 theme Sequential 741:750 arg1 degradation 752:762 Sequential degradation 741:762 Sequential degradation using partial acid hydrolysis 741:792 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 50 theme -galacturono-oligosaccharide 958:985 arg1 chain 992:996 an α-(1→4)-galacturono-oligosaccharide main chain 948:996 an α-(1→4)-galacturono-oligosaccharide main chain 948:996 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 5 51 theme ESI-MS/MS 721:729 arg1 analyses 731:738 ESI-MS/MS analyses 721:738 ESI-MS/MS analyses 721:738 The primary structure was elucidated based on sugar composition, methylation analysis, oligosaccharide analysis, and sequencing using GC, GC-MS, LC-MS, and ESI-MS/MS analyses. 27693835 6 52 contain contained 816:824 arg2 nonasaccharide 877:890 an AceA-containing nonasaccharide 858:890 an AceA-containing nonasaccharide 858:890 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 52 contain contained 816:824 arg2 -Kdo 836:839 Rhap-(1→5)-Kdo 826:839 Rhap-(1→5)-Kdo 826:839 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 52 contain contained 816:824 arg2 -Dha 852:855 Araf-(1→5)-Dha 842:855 Araf-(1→5)-Dha 842:855 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 52 contain contained 816:824 arg2 oligosaccharide 917:931 an uronic acid-rich oligosaccharide 897:931 an uronic acid-rich oligosaccharide 897:931 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 6 52 contain contained 816:824 arg1 CPE-II 809:814 CPE-II 809:814 CPE-II 809:814 Sequential degradation using partial acid hydrolysis indicated that CPE-II contained Rhap-(1→5)-Kdo, Araf-(1→5)-Dha, an AceA-containing nonasaccharide, and an uronic acid-rich oligosaccharide in addition to an α-(1→4)-galacturono-oligosaccharide main chain. 27693835 1 53 theme anti-metastatic 212:226 arg1 property 228:235 an anti-metastatic property 209:235 an anti-metastatic property 209:235 The aim of this study was to characterize a polysaccharide found in citrus peels with an anti-metastatic property. 27693835 3 54 theme lung 317:320 arg1 metastasis 322:331 in vivo lung metastasis 309:331 in vivo lung metastasis of Colon26-M3.1 309:347 During in vivo lung metastasis of Colon26-M3.1, administration of 10μg of CPE-II per mouse showed 81.3% inhibition of metastasis. 26179816 0 0 theme Acid 109:112 arg1 Esterification 114:127 Linkage-Specific Sialic Acid Esterification 85:127 Linkage-Specific Sialic Acid Esterification 85:127 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 5 1 theme first 818:822 arg1 samples 834:840 96 samples 831:840 96 samples 831:840 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 5 1 theme first 818:822 arg1 set 824:826 a first set 816:826 a first set of 96 samples 816:840 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 6 2 theme thus-obtained 929:941 arg1 glycans 943:949 the thus-obtained glycans 925:949 the thus-obtained glycans 925:949 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 0 3 theme Sialic 102:107 arg1 Esterification 114:127 Linkage-Specific Sialic Acid Esterification 85:127 Linkage-Specific Sialic Acid Esterification 85:127 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 1 4 theme heterogeneous 204:216 arg1 characteristics 229:243 heterogeneous structural characteristics 204:243 heterogeneous structural characteristics 204:243 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 4 5 from automation 655:664 arg1 system 706:711 a liquid-handling robot system 682:711 a liquid-handling robot system 682:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 4 6 theme current 588:594 arg1 study 596:600 the current study 584:600 the current study 584:600 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 4 7 used used 622:625 arg2 point 641:645 a starting point 630:645 a starting point for the automation of all steps on a liquid-handling robot system 630:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 4 7 used used 622:625 arg2 procedure 608:616 this procedure 603:616 this procedure 603:616 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 2 8 theme human 357:361 arg1 N-glycans 370:378 human plasma N-glycans 357:378 human plasma N-glycans 357:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 5 9 with procedure 771:779 arg1 times 797:801 throughput times 786:801 throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 786:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 6 10 theme interday 1026:1033 arg1 repeatability 1035:1047 improved interday repeatability 1017:1047 improved interday repeatability 1017:1047 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 3 11 theme acid 406:409 arg1 residues 411:418 sialic acid residues 399:418 sialic acid residues 399:418 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 1 12 with modification 168:179 arg1 characteristics 229:243 heterogeneous structural characteristics 204:243 heterogeneous structural characteristics 204:243 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 1 13 theme structural 218:227 arg1 characteristics 229:243 heterogeneous structural characteristics 204:243 heterogeneous structural characteristics 204:243 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 5 14 theme h 810:810 arg1 times 797:801 throughput times 786:801 throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 786:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 2 15 theme comprehensive 330:342 arg1 profiling 344:352 the comprehensive profiling 326:352 the comprehensive profiling of human plasma N-glycans 326:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 3 16 theme sialic 499:504 arg1 residues 511:518 α2,6-linked sialic acid residues 487:518 α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids 487:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 6 17 theme improved 1017:1024 arg1 repeatability 1035:1047 improved interday repeatability 1017:1047 improved interday repeatability 1017:1047 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 3 18 theme acid 506:509 arg1 residues 511:518 α2,6-linked sialic acid residues 487:518 α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids 487:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 5 19 theme h 862:862 arg1 times 797:801 throughput times 786:801 throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 786:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 5 20 theme standardized 758:769 arg1 procedure 771:779 a time-efficient and fully standardized procedure 731:779 a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 731:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 2 21 theme N-glycans 370:378 arg1 profiling 344:352 the comprehensive profiling 326:352 the comprehensive profiling of human plasma N-glycans 326:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 2 22 dep robust 278:283 arg1 high-throughput 286:300 high-throughput 286:300 high-throughput 286:300 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 5 23 theme extra 864:868 arg1 h 862:862 approximately 1 h extra 846:868 approximately 1 h extra for each additional sample plate 846:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 4 24 theme starting 632:639 arg1 procedure 608:616 this procedure 603:616 this procedure 603:616 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 4 24 theme starting 632:639 arg1 point 641:645 a starting point 630:645 a starting point for the automation of all steps on a liquid-handling robot system 630:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 6 25 theme mass 908:911 arg1 analysis 913:920 The mass analysis 904:920 The mass analysis of the thus-obtained glycans 904:949 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 6 25 theme mass 908:911 arg1 reproducible 962:973 reproducible 962:973 reproducible 962:973 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 2 26 theme plasma 363:368 arg1 N-glycans 370:378 human plasma N-glycans 357:378 human plasma N-glycans 357:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 0 27 theme Plasma 54:59 arg1 Analysis 71:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 6 28 theme quantification 996:1009 arg1 terms 978:982 terms 978:982 terms of relative quantification 978:1009 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 5 29 theme additional 879:888 arg1 plate 897:901 each additional sample plate 874:901 each additional sample plate 874:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 5 30 theme throughput 786:795 arg1 times 797:801 throughput times 786:801 throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 786:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 3 31 theme parallel 525:532 arg1 formation 542:550 parallel lactone formation 525:550 parallel lactone formation of α2,3-linked sialic acids 525:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 0 32 theme Spectrometry-Based 35:52 arg1 Analysis 71:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 6 33 theme relative 987:994 arg1 quantification 996:1009 relative quantification 987:1009 relative quantification 987:1009 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 3 34 theme α2,6-linked 487:497 arg1 residues 511:518 α2,6-linked sialic acid residues 487:518 α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids 487:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 5 35 theme sample 890:895 arg1 plate 897:901 each additional sample plate 874:901 each additional sample plate 874:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 3 36 theme lactone 534:540 arg1 formation 542:550 parallel lactone formation 525:550 parallel lactone formation of α2,3-linked sialic acids 525:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 0 37 theme Analysis 71:78 arg1 Automation 0:9 Automation 0:9 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 0:128 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 3 38 link α2,3-linked 555:565 arg1 acids 574:578 α2,3-linked sialic acids 555:578 α2,3-linked sialic acids 555:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 3 39 theme residues 511:518 arg1 ethylation 473:482 the ethylation 469:482 the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids 469:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 3 39 theme residues 511:518 arg1 linkage-specificity 441:459 linkage-specificity 441:459 linkage-specificity 441:459 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 2 40 theme robust 278:283 arg1 method 315:320 a robust, high-throughput MALDI-TOF-MS method 276:320 a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans 276:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 3 41 theme acids 574:578 arg1 formation 542:550 parallel lactone formation 525:550 parallel lactone formation of α2,3-linked sialic acids 525:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 0 42 theme N-Glycome 61:69 arg1 Analysis 71:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis 14:78 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 5 43 theme time-efficient 733:746 arg1 procedure 771:779 a time-efficient and fully standardized procedure 731:779 a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate 731:901 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 3 44 link α2,6-linked 487:497 arg1 residues 511:518 α2,6-linked sialic acid residues 487:518 α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids 487:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 6 45 from terms 978:982 arg1 analysis 913:920 The mass analysis 904:920 The mass analysis of the thus-obtained glycans 904:949 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 6 45 from terms 978:982 arg1 reproducible 962:973 reproducible 962:973 reproducible 962:973 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 1 46 theme post-translational 149:166 arg1 modification 168:179 a post-translational modification 147:179 a post-translational modification of key importance with heterogeneous structural characteristics 147:243 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 1 46 theme post-translational 149:166 arg1 Glycosylation 130:142 Glycosylation 130:142 Glycosylation 130:142 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 3 47 with residues 511:518 arg1 formation 542:550 parallel lactone formation 525:550 parallel lactone formation of α2,3-linked sialic acids 525:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 5 48 theme samples 834:840 arg1 samples 834:840 96 samples 831:840 96 samples 831:840 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 5 48 theme samples 834:840 arg1 set 824:826 a first set 816:826 a first set of 96 samples 816:840 This resulted in a time-efficient and fully standardized procedure with throughput times of 2.5 h for a first set of 96 samples and approximately 1 h extra for each additional sample plate. 26179816 0 49 with Automation 0:9 arg1 Esterification 114:127 Linkage-Specific Sialic Acid Esterification 85:127 Linkage-Specific Sialic Acid Esterification 85:127 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 3 50 theme α2,3-linked 555:565 arg1 acids 574:578 α2,3-linked sialic acids 555:578 α2,3-linked sialic acids 555:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 6 51 theme manual 1072:1077 arg1 processing 1079:1088 manual processing 1072:1088 manual processing 1072:1088 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 3 52 theme sialic 567:572 arg1 acids 574:578 α2,3-linked sialic acids 555:578 α2,3-linked sialic acids 555:578 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 6 53 with reproducible 962:973 arg1 repeatability 1035:1047 improved interday repeatability 1017:1047 improved interday repeatability 1017:1047 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 0 54 theme Linkage-Specific 85:100 arg1 Esterification 114:127 Linkage-Specific Sialic Acid Esterification 85:127 Linkage-Specific Sialic Acid Esterification 85:127 Automation of High-Throughput Mass Spectrometry-Based Plasma N-Glycome Analysis with Linkage-Specific Sialic Acid Esterification. 26179816 4 55 theme robot 700:704 arg1 system 706:711 a liquid-handling robot system 682:711 a liquid-handling robot system 682:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 4 56 theme liquid-handling 684:698 arg1 system 706:711 a liquid-handling robot system 682:711 a liquid-handling robot system 682:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 26179816 1 57 theme key 184:186 arg1 importance 188:197 key importance 184:197 key importance 184:197 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 6 58 theme glycans 943:949 arg1 analysis 913:920 The mass analysis 904:920 The mass analysis of the thus-obtained glycans 904:949 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 6 58 theme glycans 943:949 arg1 reproducible 962:973 reproducible 962:973 reproducible 962:973 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 3 59 theme sialic 399:404 arg1 residues 411:418 sialic acid residues 399:418 sialic acid residues 399:418 In this approach, sialic acid residues are derivatized with linkage-specificity, namely the ethylation of α2,6-linked sialic acid residues with parallel lactone formation of α2,3-linked sialic acids. 26179816 2 60 theme MALDI-TOF-MS 302:313 arg1 method 315:320 a robust, high-throughput MALDI-TOF-MS method 276:320 a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans 276:378 Previously, we have developed a robust, high-throughput MALDI-TOF-MS method for the comprehensive profiling of human plasma N-glycans. 26179816 1 61 theme importance 188:197 arg1 modification 168:179 a post-translational modification 147:179 a post-translational modification of key importance with heterogeneous structural characteristics 147:243 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 1 61 theme importance 188:197 arg1 Glycosylation 130:142 Glycosylation 130:142 Glycosylation 130:142 Glycosylation is a post-translational modification of key importance with heterogeneous structural characteristics. 26179816 6 62 from reproducible 962:973 arg1 terms 978:982 terms 978:982 terms of relative quantification 978:1009 The mass analysis of the thus-obtained glycans was highly reproducible in terms of relative quantification, with improved interday repeatability as compared to that of manual processing. 26179816 4 63 theme steps 673:677 arg1 automation 655:664 the automation 651:664 the automation of all steps on a liquid-handling robot system 651:711 In the current study, this procedure was used as a starting point for the automation of all steps on a liquid-handling robot system. 28842505 7 0 theme receptor 1251:1258 arg1 redirection 1226:1236 the redirection 1222:1236 the redirection of the Notch receptor to the lysosome 1222:1274 The inhibition resulted from the redirection of the Notch receptor to the lysosome, a novel mechanism. 28842505 5 1 theme Notch 917:921 arg1 receptor 923:930 the heavily glycosylated Notch receptor 892:930 the heavily glycosylated Notch receptor involved in normal and malignant development 892:975 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 6 2 theme primary 1023:1029 arg1 cells 1045:1049 primary tumor-derived cells 1023:1049 primary tumor-derived cells 1023:1049 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 2 3 theme forms 505:509 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 3 4 from study 520:524 arg1 interested 534:543 interested 534:543 interested 534:543 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 5 5 theme receptor 923:930 arg1 activation 878:887 activation 878:887 activation 878:887 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 5 5 theme receptor 923:930 arg1 levels 867:872 levels 867:872 levels 867:872 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 5 6 dep levels 867:872 arg1 the 863:865 the 863:865 the 863:865 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 2 7 link N-linked 446:453 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 4 8 theme cancer 782:787 arg1 cells 794:798 cancer stem cells 782:798 cancer stem cells 782:798 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 6 9 theme γ-secretase 1169:1179 arg1 inhibition 1181:1190 γ-secretase inhibition 1169:1190 γ-secretase inhibition 1169:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 7 10 theme Notch 1245:1249 arg1 receptor 1251:1258 the Notch receptor 1241:1258 the Notch receptor 1241:1258 The inhibition resulted from the redirection of the Notch receptor to the lysosome, a novel mechanism. 28842505 6 11 link tumor-derived 1031:1043 arg1 cells 1045:1049 primary tumor-derived cells 1023:1049 primary tumor-derived cells 1023:1049 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 6 12 theme cell 1008:1011 arg1 lines 1013:1017 EOC cell lines 1004:1017 EOC cell lines 1004:1017 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 8 13 theme new 1325:1327 arg1 role 1329:1332 a new role 1323:1332 a new role for bisecting glycosylation in the control of Notch transport 1323:1394 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 2 14 theme gene 292:295 arg1 Mgat3 297:301 the gene Mgat3 288:301 the gene Mgat3 encoding the glycosyltransferase GnT-III 288:342 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 4 15 theme shRNA 672:676 arg1 suppression 683:693 stable shRNA gene suppression 665:693 stable shRNA gene suppression 665:693 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 6 16 theme Notch 1148:1152 arg1 activation 1154:1163 Notch activation 1148:1163 Notch activation via γ-secretase inhibition 1148:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 4 17 theme stem 789:792 arg1 cells 794:798 cancer stem cells 782:798 cancer stem cells 782:798 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 1 18 theme cellular 181:188 arg1 transformation 190:203 cellular transformation 181:203 cellular transformation 181:203 Glycosylation changes associated with cellular transformation can facilitate the growth and progression of tumors. 28842505 0 19 theme glycosyltransferase 4:22 arg1 GnT-III 24:30 The glycosyltransferase GnT-III 0:30 The glycosyltransferase GnT-III 0:30 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 5 20 theme malignant 955:963 arg1 development 965:975 normal and malignant development 944:975 normal and malignant development 944:975 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 2 21 theme structures 462:471 arg1 production 413:422 the production 409:422 the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms 409:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 6 22 theme EOC 1004:1006 arg1 lines 1013:1017 EOC cell lines 1004:1017 EOC cell lines 1004:1017 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 4 23 theme cells 761:765 arg1 expansion 732:740 the expansion 728:740 the expansion of side-population cells 728:765 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 4 24 theme gene 678:681 arg1 suppression 683:693 stable shRNA gene suppression 665:693 stable shRNA gene suppression 665:693 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 2 25 theme glycan 455:460 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 26 theme glycosyltransferase 316:334 arg1 GnT-III 336:342 the glycosyltransferase GnT-III 312:342 the glycosyltransferase GnT-III 312:342 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 27 theme truncated 436:444 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 28 theme N-linked 446:453 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 6 29 theme GnT-III 993:999 arg1 Suppression 978:988 Suppression 978:988 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells 978:1049 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 8 30 theme transport 1386:1394 arg1 control 1369:1375 the control 1365:1375 the control of Notch transport 1365:1394 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 8 31 theme EOC 1491:1493 arg1 growth 1495:1500 EOC growth 1491:1500 EOC growth 1491:1500 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 3 32 theme abnormal 570:577 arg1 glycans 579:585 these abnormal glycans 564:585 these abnormal glycans 564:585 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 2 33 theme abnormal 427:434 arg1 structures 462:471 abnormal truncated N-linked glycan structures 427:471 abnormal truncated N-linked glycan structures instead of the typical bisected forms 427:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 34 theme bisected 496:503 arg1 forms 505:509 the typical bisected forms 484:509 the typical bisected forms 484:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 6 35 theme activation 1154:1163 arg1 blockage 1136:1143 pharmacologic blockage 1122:1143 pharmacologic blockage of Notch activation via γ-secretase inhibition 1122:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 0 36 theme cancer 135:140 arg1 invasion 115:122 invasion 115:122 invasion 115:122 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 0 36 theme cancer 135:140 arg1 growth 104:109 growth 104:109 growth 104:109 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 7 37 theme novel 1279:1283 arg1 mechanism 1285:1293 a novel mechanism 1277:1293 a novel mechanism 1277:1293 The inhibition resulted from the redirection of the Notch receptor to the lysosome, a novel mechanism. 28842505 6 38 theme signaling 1086:1094 arg1 inhibition 1066:1075 an inhibition 1063:1075 an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition 1063:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 2 39 theme typical 488:494 arg1 forms 505:509 the typical bisected forms 484:509 the typical bisected forms 484:509 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 0 40 theme Notch 42:46 arg1 signaling 48:56 Notch signaling 42:56 Notch signaling 42:56 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 8 41 theme bisecting 1338:1346 arg1 glycosylation 1348:1360 bisecting glycosylation 1338:1360 bisecting glycosylation 1338:1360 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 5 42 gly glycosylated 904:915 arg1 receptor 923:930 the heavily glycosylated Notch receptor 892:930 the heavily glycosylated Notch receptor involved in normal and malignant development 892:975 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 8 43 theme GnT-III 1452:1458 arg1 potential 1428:1436 the therapeutic potential 1412:1436 the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence 1412:1515 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 2 44 theme ovarian 370:376 arg1 EOCs 390:393 EOCs 390:393 EOCs 390:393 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 44 theme ovarian 370:376 arg1 carcinomas 378:387 epithelial ovarian carcinomas 359:387 epithelial ovarian carcinomas (EOCs) 359:394 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 0 45 theme ovarian 127:133 arg1 cancer 135:140 ovarian cancer 127:140 ovarian cancer 127:140 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 8 46 theme inhibiting 1441:1450 arg1 GnT-III 1452:1458 inhibiting GnT-III 1441:1458 inhibiting GnT-III 1441:1458 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 2 47 theme epithelial 359:368 arg1 EOCs 390:393 EOCs 390:393 EOCs 390:393 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 2 47 theme epithelial 359:368 arg1 carcinomas 378:387 epithelial ovarian carcinomas 359:387 epithelial ovarian carcinomas (EOCs) 359:394 Previously we discovered that the gene Mgat3 encoding the glycosyltransferase GnT-III is elevated in epithelial ovarian carcinomas (EOCs) and leads to the production of abnormal truncated N-linked glycan structures instead of the typical bisected forms. 28842505 6 48 from Suppression 978:988 arg1 cells 1045:1049 primary tumor-derived cells 1023:1049 primary tumor-derived cells 1023:1049 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 6 48 from Suppression 978:988 arg1 lines 1013:1017 EOC cell lines 1004:1017 EOC cell lines 1004:1017 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 3 49 theme cancer 632:637 arg1 progression 609:619 progression 609:619 progression 609:619 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 3 49 theme cancer 632:637 arg1 growth 598:603 growth 598:603 growth 598:603 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 8 50 from role 1329:1332 arg1 control 1369:1375 the control 1365:1375 the control of Notch transport 1365:1394 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 4 51 theme side-population 745:759 arg1 cells 761:765 side-population cells 745:765 side-population cells 745:765 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 8 52 theme therapeutic 1416:1426 arg1 potential 1428:1436 the therapeutic potential 1412:1436 the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence 1412:1515 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 8 53 theme Notch 1380:1384 arg1 transport 1386:1394 Notch transport 1380:1394 Notch transport 1380:1394 These findings demonstrate a new role for bisecting glycosylation in the control of Notch transport and demonstrate the therapeutic potential of inhibiting GnT-III as a treatment for controlling EOC growth and recurrence. 28842505 5 54 theme GnT-III 834:840 arg1 expression 842:851 GnT-III expression 834:851 GnT-III expression 834:851 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 5 55 theme glycosylated 904:915 arg1 receptor 923:930 the heavily glycosylated Notch receptor 892:930 the heavily glycosylated Notch receptor involved in normal and malignant development 892:975 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 5 56 theme normal 944:949 arg1 development 965:975 normal and malignant development 944:975 normal and malignant development 944:975 More specifically, we found that GnT-III expression regulates the levels and activation of the heavily glycosylated Notch receptor involved in normal and malignant development. 28842505 0 57 theme cell 74:77 arg1 expansion 79:87 stem cell expansion 69:87 stem cell expansion to promote the growth and invasion of ovarian cancer 69:140 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 6 58 theme pharmacologic 1122:1134 arg1 blockage 1136:1143 pharmacologic blockage 1122:1143 pharmacologic blockage of Notch activation via γ-secretase inhibition 1122:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 6 59 theme Notch 1080:1084 arg1 signaling 1086:1094 Notch signaling 1080:1094 Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition 1080:1190 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 28842505 4 60 theme GnT-III 700:706 arg1 expression 708:717 GnT-III expression 700:717 GnT-III expression 700:717 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 4 61 theme stable 665:670 arg1 suppression 683:693 stable shRNA gene suppression 665:693 stable shRNA gene suppression 665:693 We have discovered using stable shRNA gene suppression that GnT-III expression controls the expansion of side-population cells, also known as cancer stem cells. 28842505 1 62 theme tumors 250:255 arg1 progression 235:245 progression 235:245 progression 235:245 Glycosylation changes associated with cellular transformation can facilitate the growth and progression of tumors. 28842505 1 62 theme tumors 250:255 arg1 growth 224:229 growth 224:229 growth 224:229 Glycosylation changes associated with cellular transformation can facilitate the growth and progression of tumors. 28842505 0 63 theme stem 69:72 arg1 expansion 79:87 stem cell expansion 69:87 stem cell expansion to promote the growth and invasion of ovarian cancer 69:140 The glycosyltransferase GnT-III activates Notch signaling and drives stem cell expansion to promote the growth and invasion of ovarian cancer. 28842505 3 64 theme ovarian 624:630 arg1 cancer 632:637 ovarian cancer 624:637 ovarian cancer 624:637 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 1 65 theme Glycosylation 143:155 arg1 changes 157:163 Glycosylation changes 143:163 Glycosylation changes associated with cellular transformation 143:203 Glycosylation changes associated with cellular transformation can facilitate the growth and progression of tumors. 28842505 3 66 from interested 534:543 arg1 study 520:524 this study 515:524 this study 515:524 In this study, we are interested in discovering how these abnormal glycans impact the growth and progression of ovarian cancer. 28842505 6 67 theme tumor-derived 1031:1043 arg1 cells 1045:1049 primary tumor-derived cells 1023:1049 primary tumor-derived cells 1023:1049 Suppression of GnT-III in EOC cell lines and primary tumor-derived cells resulted in an inhibition of Notch signaling that was more potent than pharmacologic blockage of Notch activation via γ-secretase inhibition. 26372017 1 0 theme transglycosylation 154:171 arg1 reaction 173:180 transglycosylation reaction 154:180 transglycosylation reaction of rice seed α-glucosidase 154:207 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 1 theme glucosyl 257:264 arg1 donor 266:270 a glucosyl donor 255:270 a glucosyl donor 255:270 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 1 theme glucosyl 257:264 arg1 maltose 244:250 maltose 244:250 maltose 244:250 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 5 2 theme Rice 858:861 arg1 α-glucosidase 868:880 Rice seed α-glucosidase 858:880 Rice seed α-glucosidase 858:880 Rice seed α-glucosidase acted on maltose, soluble starch, and PNP α-D-glucopyranoside as glycosyl donors, but not on glucose, sucrose, or trehalose. 26372017 5 3 theme seed 863:866 arg1 α-glucosidase 868:880 Rice seed α-glucosidase 858:880 Rice seed α-glucosidase 858:880 Rice seed α-glucosidase acted on maltose, soluble starch, and PNP α-D-glucopyranoside as glycosyl donors, but not on glucose, sucrose, or trehalose. 26372017 3 4 from maltose 555:561 arg1 group 544:548 α-glucosyl group 533:548 α-glucosyl group from maltose to G-Rg1 533:570 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 6 5 theme G-Rg1 1043:1047 arg1 derivatives 1028:1038 These α-monoglucosyl derivatives 1007:1038 These α-monoglucosyl derivatives of G-Rg1 1007:1047 These α-monoglucosyl derivatives of G-Rg1 were easily hydrolyzed to G-Rg1 by rat small intestinal and liver α-glucosidase in vitro. 26372017 3 6 theme -glucosidic 657:667 arg1 linkages 669:676 either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages 583:676 linkages 669:676 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 6 7 theme rat 1084:1086 arg1 α-glucosidase 1115:1127 rat small intestinal and liver α-glucosidase 1084:1127 rat small intestinal and liver α-glucosidase 1084:1127 These α-monoglucosyl derivatives of G-Rg1 were easily hydrolyzed to G-Rg1 by rat small intestinal and liver α-glucosidase in vitro. 26372017 5 8 theme PNP 920:922 arg1 α-D-glucopyranoside 924:942 PNP α-D-glucopyranoside 920:942 PNP α-D-glucopyranoside as glycosyl donors 920:961 Rice seed α-glucosidase acted on maltose, soluble starch, and PNP α-D-glucopyranoside as glycosyl donors, but not on glucose, sucrose, or trehalose. 26372017 3 9 theme seed 504:507 arg1 α-glucosidase 509:521 rice seed α-glucosidase 499:521 rice seed α-glucosidase 499:521 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 6 10 dep rat 1084:1086 arg1 intestinal 1094:1103 intestinal 1094:1103 intestinal 1094:1103 These α-monoglucosyl derivatives of G-Rg1 were easily hydrolyzed to G-Rg1 by rat small intestinal and liver α-glucosidase in vitro. 26372017 1 11 theme seed 190:193 arg1 α-glucosidase 195:207 rice seed α-glucosidase 185:207 rice seed α-glucosidase 185:207 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 3 12 theme β-glucose 681:689 arg1 moieties 691:698 β-glucose moieties 681:698 β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone 681:773 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 1 13 theme α-monoglucosyl 80:93 arg1 acceptor 288:295 an acceptor 285:295 an acceptor 285:295 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 13 theme α-monoglucosyl 80:93 arg1 G-Rg1 127:131 G-Rg1 127:131 G-Rg1 127:131 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 13 theme α-monoglucosyl 80:93 arg1 derivatives 95:105 Six α-monoglucosyl derivatives 76:105 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) 76:132 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 2 14 theme time 395:398 arg1 effects 375:381 the effects 371:381 the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction 371:454 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 2 15 theme reaction 386:393 arg1 time 395:398 reaction time 386:398 reaction time 386:398 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 0 16 theme Enzymatic 0:8 arg1 transglycosylation 10:27 Enzymatic transglycosylation 0:27 Enzymatic transglycosylation of ginsenoside Rg1 by rice 0:54 Enzymatic transglycosylation of ginsenoside Rg1 by rice seed α-glucosidase. 26372017 4 17 theme transglycosylation 805:822 arg1 reaction 824:831 the transglycosylation reaction 801:831 the transglycosylation reaction 801:831 The optimum pH range for the transglycosylation reaction was between 5.0 and 6.0. 26372017 2 18 from effects 375:381 arg1 reaction 447:454 transglycosylation reaction 428:454 transglycosylation reaction 428:454 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 5 19 theme soluble 900:906 arg1 starch 908:913 soluble starch 900:913 soluble starch 900:913 Rice seed α-glucosidase acted on maltose, soluble starch, and PNP α-D-glucopyranoside as glycosyl donors, but not on glucose, sucrose, or trehalose. 26372017 1 20 theme α-glucosidase 195:207 arg1 reaction 173:180 transglycosylation reaction 154:180 transglycosylation reaction of rice seed α-glucosidase 154:207 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 0 21 theme ginsenoside 32:42 arg1 Rg1 44:46 ginsenoside Rg1 32:46 ginsenoside Rg1 32:46 Enzymatic transglycosylation of ginsenoside Rg1 by rice seed α-glucosidase. 26372017 6 22 theme α-monoglucosyl 1013:1026 arg1 derivatives 1028:1038 These α-monoglucosyl derivatives 1007:1038 These α-monoglucosyl derivatives of G-Rg1 1007:1047 These α-monoglucosyl derivatives of G-Rg1 were easily hydrolyzed to G-Rg1 by rat small intestinal and liver α-glucosidase in vitro. 26372017 4 23 theme pH 788:789 arg1 range 791:795 The optimum pH range 776:795 The optimum pH range for the transglycosylation reaction 776:831 The optimum pH range for the transglycosylation reaction was between 5.0 and 6.0. 26372017 2 24 theme chemical 304:311 arg1 structures 313:322 Their chemical structures 298:322 Their chemical structures 298:322 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 2 25 theme glycosyl 409:416 arg1 donors 418:423 glycosyl donors 409:423 glycosyl donors 409:423 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 2 26 theme donors 418:423 arg1 effects 375:381 the effects 371:381 the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction 371:454 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 2 27 theme transglycosylation 428:445 arg1 reaction 447:454 transglycosylation reaction 428:454 transglycosylation reaction 428:454 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 1 28 theme Rg1 122:124 arg1 acceptor 288:295 an acceptor 285:295 an acceptor 285:295 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 28 theme Rg1 122:124 arg1 G-Rg1 127:131 G-Rg1 127:131 G-Rg1 127:131 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 28 theme Rg1 122:124 arg1 derivatives 95:105 Six α-monoglucosyl derivatives 76:105 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) 76:132 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 3 29 theme rice 499:502 arg1 α-glucosidase 509:521 rice seed α-glucosidase 499:521 rice seed α-glucosidase 499:521 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 4 30 theme optimum 780:786 arg1 range 791:795 The optimum pH range 776:795 The optimum pH range for the transglycosylation reaction 776:831 The optimum pH range for the transglycosylation reaction was between 5.0 and 6.0. 26372017 1 31 theme reaction 216:223 arg1 mixture 225:231 the reaction mixture 212:231 the reaction mixture containing maltose as a glucosyl donor and G-Rg1 212:280 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 0 32 theme Rg1 44:46 arg1 transglycosylation 10:27 Enzymatic transglycosylation 0:27 Enzymatic transglycosylation of ginsenoside Rg1 by rice 0:54 Enzymatic transglycosylation of ginsenoside Rg1 by rice seed α-glucosidase. 26372017 0 33 gly transglycosylation 10:27 arg1 Rg1 44:46 ginsenoside Rg1 32:46 ginsenoside Rg1 32:46 Enzymatic transglycosylation of ginsenoside Rg1 by rice seed α-glucosidase. 26372017 2 34 theme pH 401:402 arg1 effects 375:381 the effects 371:381 the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction 371:454 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 3 35 from linkages 669:676 arg1 moieties 691:698 β-glucose moieties 681:698 β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone 681:773 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 36 theme α-1,3 590:594 arg1 linkages 669:676 either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages 583:676 linkages 669:676 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 1 37 contain containing 233:242 arg2 donor 266:270 a glucosyl donor 255:270 a glucosyl donor 255:270 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 37 contain containing 233:242 arg2 maltose 244:250 maltose 244:250 maltose 244:250 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 1 37 contain containing 233:242 arg1 mixture 225:231 the reaction mixture 212:231 the reaction mixture containing maltose as a glucosyl donor and G-Rg1 212:280 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 2 38 theme spectroscopic 343:355 arg1 analysis 357:364 spectroscopic analysis 343:364 spectroscopic analysis 343:364 Their chemical structures were identified by spectroscopic analysis, and the effects of reaction time, pH, and glycosyl donors on transglycosylation reaction were investigated. 26372017 5 39 theme glycosyl 947:954 arg1 donors 956:961 glycosyl donors 947:961 glycosyl donors 947:961 Rice seed α-glucosidase acted on maltose, soluble starch, and PNP α-D-glucopyranoside as glycosyl donors, but not on glucose, sucrose, or trehalose. 26372017 3 40 theme α-1,4 612:616 arg1 linkages 669:676 either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages 583:676 linkages 669:676 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 1 41 theme ginsenoside 110:120 arg1 Rg1 122:124 ginsenoside Rg1 110:124 ginsenoside Rg1 110:124 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 3 42 theme aglycone 766:773 arg1 C20-position 722:733 C20-position 722:733 C20-position 722:733 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 42 theme aglycone 766:773 arg1 C6- 714:716 the C6- 710:716 the C6- 710:716 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 6 43 theme liver 1109:1113 arg1 α-glucosidase 1115:1127 rat small intestinal and liver α-glucosidase 1084:1127 rat small intestinal and liver α-glucosidase 1084:1127 These α-monoglucosyl derivatives of G-Rg1 were easily hydrolyzed to G-Rg1 by rat small intestinal and liver α-glucosidase in vitro. 26372017 3 44 theme α-1,6 636:640 arg1 linkages 669:676 either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages 583:676 linkages 669:676 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 1 45 theme rice 185:188 arg1 α-glucosidase 195:207 rice seed α-glucosidase 185:207 rice seed α-glucosidase 185:207 Six α-monoglucosyl derivatives of ginsenoside Rg1 (G-Rg1) were synthesized by transglycosylation reaction of rice seed α-glucosidase in the reaction mixture containing maltose as a glucosyl donor and G-Rg1 as an acceptor. 26372017 3 46 attach linked 700:705 arg2 moieties 691:698 β-glucose moieties 681:698 β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone 681:773 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 46 attach linked 700:705 arg3 C6- 714:716 the C6- 710:716 the C6- 710:716 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 46 attach linked 700:705 arg3 C20-position 722:733 C20-position 722:733 C20-position 722:733 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 47 theme α-glucosyl 533:542 arg1 group 544:548 α-glucosyl group 533:548 α-glucosyl group from maltose to G-Rg1 533:570 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26372017 3 48 theme -type 760:764 arg1 aglycone 766:773 protopanaxatriol (PPT)-type aglycone 738:773 protopanaxatriol (PPT)-type aglycone 738:773 The results showed that rice seed α-glucosidase transfers α-glucosyl group from maltose to G-Rg1 by forming either α-1,3 (α-nigerosyl)-, α-1,4 (α-maltosyl)-, or α-1,6 (α-isomaltosyl)-glucosidic linkages in β-glucose moieties linked at the C6- and C20-position of protopanaxatriol (PPT)-type aglycone. 26304114 6 0 from substrate 903:911 arg1 site 927:930 the active site 916:930 the active site 916:930 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 7 1 theme catalytic 1153:1161 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 1 theme catalytic 1153:1161 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 1 theme catalytic 1153:1161 arg1 distant 1201:1207 distant 1201:1207 distant 1201:1207 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 8 2 theme catalytic 1463:1471 arg1 acid/base 1473:1481 the catalytic acid/base 1459:1481 the catalytic acid/base 1459:1481 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 6 3 theme lid 881:883 arg1 closing 865:871 The closing 861:871 The closing of this lid 861:883 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 8 4 theme same 1366:1369 arg1 complexes 1371:1379 These same complexes 1360:1379 These same complexes 1360:1379 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 7 5 theme acid/base 1163:1171 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 5 theme acid/base 1163:1171 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 5 theme acid/base 1163:1171 arg1 distant 1201:1207 distant 1201:1207 distant 1201:1207 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 0 6 from Analysis 11:18 arg1 Complex 59:65 Complex 59:65 Complex with Carbohydrates 59:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 5 7 dep residues 814:821 arg1 724-WNW-726 823:833 724-WNW-726 823:833 724-WNW-726 823:833 Upon substrate binding, a tryptophan lid (residues 724-WNW-726) closes on the substrate. 26304114 7 8 located found 1185:1189 arg1 forms 1110:1114 the bound forms 1100:1114 the bound forms of the enzyme 1100:1128 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 8 located found 1185:1189 arg2 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 8 located found 1185:1189 arg2 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 8 located found 1185:1189 arg2 distant 1201:1207 distant 1201:1207 distant 1201:1207 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 8 located found 1185:1189 arg1 all 1093:1095 all 1093:1095 all 1093:1095 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 4 9 with complex 704:710 arg1 analogues 761:769 substrate analogues 751:769 substrate analogues 751:769 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 9 with complex 704:710 arg1 inhibitor 736:744 inhibitor 736:744 inhibitor 736:744 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 9 with complex 704:710 arg1 products 726:733 reaction products 717:733 reaction products 717:733 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 7 10 theme acid/base 1286:1294 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 10 theme acid/base 1286:1294 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 2 11 dep serine 364:369 arg1 residues 384:391 residues 384:391 residues 384:391 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 9 12 theme structural 1532:1541 arg1 observations 1543:1554 these structural observations 1526:1554 these structural observations 1526:1554 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 7 13 dep serve 1248:1252 arg1 facilitate 1316:1325 facilitate 1316:1325 facilitate cleavage of the glycosidic bond 1316:1357 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 14 theme proposed 1144:1151 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 14 theme proposed 1144:1151 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 14 theme proposed 1144:1151 arg1 distant 1201:1207 distant 1201:1207 distant 1201:1207 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 0 15 from Hydrolase 46:54 arg1 Complex 59:65 Complex 59:65 Complex with Carbohydrates 59:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 9 16 theme retaining 1590:1598 arg1 mechanism 1620:1628 the retaining glycoside hydrolase mechanism 1586:1628 the retaining glycoside hydrolase mechanism 1586:1628 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 0 17 with Complex 59:65 arg1 Carbohydrates 72:84 Carbohydrates 72:84 Carbohydrates 72:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 4 18 theme substrate 751:759 arg1 analogues 761:769 substrate analogues 751:769 substrate analogues 751:769 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 6 19 theme sugar 983:987 arg1 residue 989:995 the sugar residue 979:995 the sugar residue bound within the -1 subsite 979:1023 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 2 20 attach α-linked 352:359 arg2 residue 336:342 an N-acetylgalactosamine residue 311:342 an N-acetylgalactosamine residue that is α-linked to serine or threonine residues 311:391 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 20 attach α-linked 352:359 arg1 threonine 374:382 threonine 374:382 threonine 374:382 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 20 attach α-linked 352:359 arg2 α-linked 352:359 α-linked 352:359 α-linked 352:359 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 20 attach α-linked 352:359 arg2 core 275:278 the core 271:278 the core of these glycans 271:295 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 20 attach α-linked 352:359 arg1 serine 364:369 serine 364:369 serine 364:369 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 9 21 theme glycoside 1600:1608 arg1 mechanism 1620:1628 the retaining glycoside hydrolase mechanism 1586:1628 the retaining glycoside hydrolase mechanism 1586:1628 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 5 22 theme tryptophan 798:807 arg1 lid 809:811 a tryptophan lid 796:811 a tryptophan lid (residues 724-WNW-726) 796:834 Upon substrate binding, a tryptophan lid (residues 724-WNW-726) closes on the substrate. 26304114 5 22 theme tryptophan 798:807 arg1 residues 814:821 residues 724-WNW-726 814:833 residues 724-WNW-726 814:833 Upon substrate binding, a tryptophan lid (residues 724-WNW-726) closes on the substrate. 26304114 7 23 theme glycosidic 1343:1352 arg1 bond 1354:1357 the glycosidic bond 1339:1357 the glycosidic bond 1339:1357 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 6 24 theme proposed 1046:1053 arg1 role 1055:1058 its proposed role 1042:1058 its proposed role as the catalytic nucleophile 1042:1087 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 9 25 theme general 1787:1793 arg1 residue 1815:1821 the general acid/base catalytic residue 1783:1821 the general acid/base catalytic residue 1783:1821 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 7 26 theme bound 1104:1108 arg1 forms 1110:1114 the bound forms 1100:1114 the bound forms of the enzyme 1100:1128 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 27 theme bond 1354:1357 arg1 cleavage 1327:1334 cleavage 1327:1334 cleavage of the glycosidic bond 1327:1357 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 3 28 theme known 452:456 arg1 hydrolases 404:413 Glycoside hydrolases 394:413 Glycoside hydrolases in family 101 394:427 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 3 28 theme known 452:456 arg1 enzymes 458:464 the only known enzymes 443:464 the only known enzymes to be able to hydrolyze this glycosidic linkage 443:512 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 3 29 theme Glycoside 394:402 arg1 hydrolases 404:413 Glycoside hydrolases 394:413 Glycoside hydrolases in family 101 394:427 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 3 29 theme Glycoside 394:402 arg1 enzymes 458:464 the only known enzymes 443:464 the only known enzymes to be able to hydrolyze this glycosidic linkage 443:512 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 3 30 from hydrolases 404:413 arg1 family 418:423 family 101 418:427 family 101 418:427 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 9 31 theme mechanism 1620:1628 arg1 variation 1573:1581 a new variation 1567:1581 a new variation of the retaining glycoside hydrolase mechanism 1567:1628 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 2 32 theme N-acetylgalactosamine 314:334 arg1 residue 336:342 an N-acetylgalactosamine residue 311:342 an N-acetylgalactosamine residue that is α-linked to serine or threonine residues 311:391 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 32 theme N-acetylgalactosamine 314:334 arg1 α-linked 352:359 α-linked 352:359 α-linked 352:359 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 32 theme N-acetylgalactosamine 314:334 arg1 core 275:278 the core 271:278 the core of these glycans 271:295 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 7 33 theme enzyme 1123:1128 arg1 forms 1110:1114 the bound forms 1100:1114 the bound forms of the enzyme 1100:1128 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 1 34 theme post-translational 174:191 arg1 modifications 193:205 the most abundant post-translational modifications 156:205 the most abundant post-translational modifications of proteins 156:217 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 0 35 theme Structural 0:9 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates 0:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 7 36 theme catalytic 1276:1284 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 36 theme catalytic 1276:1284 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 1 37 theme modifications 193:205 arg1 one 149:151 one 149:151 one 149:151 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 1 37 theme modifications 193:205 arg1 modifications 193:205 the most abundant post-translational modifications 156:205 the most abundant post-translational modifications of proteins 156:217 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 9 38 theme glycosidic 1728:1737 arg1 oxygen 1739:1744 the glycosidic oxygen 1724:1744 the glycosidic oxygen 1724:1744 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 9 39 dep observations 1543:1554 arg1 the 1513:1515 the 1513:1515 the 1513:1515 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 9 39 dep observations 1543:1554 arg1 basis 1517:1521 basis 1517:1521 basis 1517:1521 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 4 40 theme domain 581:586 arg1 structures 553:562 the high-resolution structures 533:562 the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues 533:769 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 9 41 theme intervening 1642:1652 arg1 molecule 1660:1667 the intervening water molecule 1638:1667 the intervening water molecule 1638:1667 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 1 42 theme proteins 210:217 arg1 modifications 193:205 the most abundant post-translational modifications 156:205 the most abundant post-translational modifications of proteins 156:217 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 5 43 theme substrate 777:785 arg1 binding 787:793 substrate binding 777:793 substrate binding 777:793 Upon substrate binding, a tryptophan lid (residues 724-WNW-726) closes on the substrate. 26304114 4 44 theme catalytic 571:579 arg1 domain 581:586 the catalytic domain 567:586 the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues 567:769 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 9 45 theme water 1654:1658 arg1 molecule 1660:1667 the intervening water molecule 1638:1667 the intervening water molecule 1638:1667 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 6 46 with consistent 1026:1035 arg1 role 1055:1058 its proposed role 1042:1058 its proposed role as the catalytic nucleophile 1042:1087 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 4 47 from TIGR4 649:653 arg1 fragment 601:608 a fragment 599:608 a fragment of GH101 from Streptococcus pneumoniae TIGR4 599:653 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 47 from TIGR4 649:653 arg1 SpGH101 656:662 SpGH101 656:662 SpGH101 656:662 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 0 48 theme Family 25:30 arg1 Hydrolase 46:54 a Family 101 Glycoside Hydrolase 23:54 a Family 101 Glycoside Hydrolase in Complex with Carbohydrates 23:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 0 49 from Complex 59:65 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates 0:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 7 50 from oxygen 1229:1234 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 50 from oxygen 1229:1234 arg1 distant 1201:1207 distant 1201:1207 distant 1201:1207 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 9 51 dep propose 1559:1565 arg1 wherein 1630:1636 wherein 1630:1636 propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue 1559:1821 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 0 52 theme Hydrolase 46:54 arg1 Analysis 11:18 Structural Analysis 0:18 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates 0:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 6 53 theme active 920:925 arg1 site 927:930 the active site 916:930 the active site 916:930 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 2 54 theme secretory 231:239 arg1 pathway 241:247 the secretory pathway 227:247 the secretory pathway of higher eukaryotes 227:268 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 0 55 theme Glycoside 36:44 arg1 Hydrolase 46:54 a Family 101 Glycoside Hydrolase 23:54 a Family 101 Glycoside Hydrolase in Complex with Carbohydrates 23:84 Structural Analysis of a Family 101 Glycoside Hydrolase in Complex with Carbohydrates Reveals Insights into Its Mechanism. 26304114 8 56 theme glycosidic 1491:1500 arg1 oxygen 1502:1507 the glycosidic oxygen 1487:1507 the glycosidic oxygen 1487:1507 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 9 57 theme hydrolase 1610:1618 arg1 mechanism 1620:1628 the retaining glycoside hydrolase mechanism 1586:1628 the retaining glycoside hydrolase mechanism 1586:1628 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 3 58 theme glycosidic 495:504 arg1 linkage 506:512 this glycosidic linkage 490:512 this glycosidic linkage 490:512 Glycoside hydrolases in family 101 are presently the only known enzymes to be able to hydrolyze this glycosidic linkage. 26304114 9 59 theme acid/base 1795:1803 arg1 residue 1815:1821 the general acid/base catalytic residue 1783:1821 the general acid/base catalytic residue 1783:1821 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 4 60 theme high-resolution 537:551 arg1 structures 553:562 the high-resolution structures 533:562 the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues 533:769 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 9 61 theme Grotthuss 1679:1687 arg1 shuttle 1696:1702 a Grotthuss proton shuttle 1677:1702 a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen 1677:1744 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 4 62 dep Streptococcus 624:636 arg1 pneumoniae 638:647 pneumoniae 638:647 pneumoniae 638:647 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 7 63 theme general 1268:1274 arg1 residue 1296:1302 a general catalytic acid/base residue 1266:1302 a general catalytic acid/base residue 1266:1302 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 63 theme general 1268:1274 arg1 residue 1173:1179 the proposed catalytic acid/base residue 1140:1179 the proposed catalytic acid/base residue 1140:1179 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 9 64 theme new 1569:1571 arg1 variation 1573:1581 a new variation 1567:1581 a new variation of the retaining glycoside hydrolase mechanism 1567:1628 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 1 65 theme O-Linked 123:130 arg1 glycosylation 132:144 O-Linked glycosylation 123:144 O-Linked glycosylation 123:144 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 9 66 theme catalytic 1805:1813 arg1 residue 1815:1821 the general acid/base catalytic residue 1783:1821 the general acid/base catalytic residue 1783:1821 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 9 67 theme proton 1689:1694 arg1 shuttle 1696:1702 a Grotthuss proton shuttle 1677:1702 a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen 1677:1744 On the basis of these structural observations we propose a new variation of the retaining glycoside hydrolase mechanism wherein the intervening water molecule enables a Grotthuss proton shuttle between Glu-796 and the glycosidic oxygen, permitting this residue to serve as the general acid/base catalytic residue. 26304114 7 68 theme glycosidic 1218:1227 arg1 oxygen 1229:1234 the glycosidic oxygen 1214:1234 the glycosidic oxygen (>4.3 Å) 1214:1243 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 7 68 theme glycosidic 1218:1227 arg1 Å 1242:1242 >4.3 Å 1237:1242 >4.3 Å 1237:1242 In all of the bound forms of the enzyme, however, the proposed catalytic acid/base residue was found to be too distant from the glycosidic oxygen (>4.3 Å) to serve directly as a general catalytic acid/base residue and thereby facilitate cleavage of the glycosidic bond. 26304114 2 69 theme glycans 289:295 arg1 core 275:278 the core 271:278 the core of these glycans 271:295 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 69 theme glycans 289:295 arg1 α-linked 352:359 α-linked 352:359 α-linked 352:359 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 2 69 theme glycans 289:295 arg1 residue 336:342 an N-acetylgalactosamine residue 311:342 an N-acetylgalactosamine residue that is α-linked to serine or threonine residues 311:391 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 8 70 theme conserved 1415:1423 arg1 molecule 1431:1438 a structurally conserved water molecule 1400:1438 a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen 1400:1507 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 4 71 theme reaction 717:724 arg1 products 726:733 reaction products 717:733 reaction products 717:733 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 72 theme GH101 613:617 arg1 fragment 601:608 a fragment 599:608 a fragment of GH101 from Streptococcus pneumoniae TIGR4 599:653 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 72 theme GH101 613:617 arg1 SpGH101 656:662 SpGH101 656:662 SpGH101 656:662 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 8 73 theme positioned 1440:1449 arg1 molecule 1431:1438 a structurally conserved water molecule 1400:1438 a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen 1400:1507 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 2 74 theme eukaryotes 259:268 arg1 pathway 241:247 the secretory pathway 227:247 the secretory pathway of higher eukaryotes 227:268 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 6 75 theme catalytic 1067:1075 arg1 nucleophile 1077:1087 the catalytic nucleophile 1063:1087 the catalytic nucleophile 1063:1087 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 1 76 theme abundant 165:172 arg1 modifications 193:205 the most abundant post-translational modifications 156:205 the most abundant post-translational modifications of proteins 156:217 O-Linked glycosylation is one of the most abundant post-translational modifications of proteins. 26304114 6 77 theme residue 989:995 arg1 C1 973:974 C1 973:974 C1 of the sugar residue bound within the -1 subsite 973:1023 The closing of this lid fully engages the substrate in the active site with Asp-764 positioned directly beneath C1 of the sugar residue bound within the -1 subsite, consistent with its proposed role as the catalytic nucleophile. 26304114 8 78 theme water 1425:1429 arg1 molecule 1431:1438 a structurally conserved water molecule 1400:1438 a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen 1400:1507 These same complexes, however, revealed a structurally conserved water molecule positioned between the catalytic acid/base and the glycosidic oxygen. 26304114 2 79 theme higher 252:257 arg1 eukaryotes 259:268 higher eukaryotes 252:268 higher eukaryotes 252:268 Within the secretory pathway of higher eukaryotes, the core of these glycans is frequently an N-acetylgalactosamine residue that is α-linked to serine or threonine residues. 26304114 4 80 theme carbohydrate 683:694 arg1 absence 672:678 the absence 668:678 the absence of carbohydrate 668:694 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 26304114 4 81 theme Streptococcus 624:636 arg1 TIGR4 649:653 Streptococcus pneumoniae TIGR4 624:653 Streptococcus pneumoniae TIGR4 624:653 Here we determine the high-resolution structures of the catalytic domain comprising a fragment of GH101 from Streptococcus pneumoniae TIGR4, SpGH101, in the absence of carbohydrate, and in complex with reaction products, inhibitor, and substrate analogues. 25064132 10 0 theme good 1137:1140 arg1 acceptors 1142:1150 good acceptors 1137:1150 good acceptors for transglycosylation 1137:1173 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 9 1 theme mM 860:861 arg1 value 846:850 a K m value 840:850 a K m value of 1.45 mM 840:861 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 2 2 theme α-galactosidase 127:141 arg1 gene 143:146 An α-galactosidase gene 124:146 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 124:188 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 2 2 theme α-galactosidase 127:141 arg1 gal36A4 149:155 gal36A4 149:155 gal36A4 149:155 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 11 3 theme diverse 1199:1205 arg1 applications 1207:1218 diverse applications 1199:1218 diverse applications in industrial fields 1199:1239 Thus, Gal36A4 may find diverse applications in industrial fields, especially in the food industry. 25064132 11 4 theme food 1260:1263 arg1 industry 1265:1272 the food industry 1256:1272 the food industry 1256:1272 Thus, Gal36A4 may find diverse applications in industrial fields, especially in the food industry. 25064132 9 5 contain had 836:838 arg1 It 787:788 It 787:788 It 787:788 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 9 5 contain had 836:838 arg2 value 846:850 a K m value 840:850 a K m value of 1.45 mM 840:861 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 5 6 theme typical 395:401 arg1 organization 408:419 the typical GH36 organization 391:419 the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet 391:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 7 7 theme pH 708:709 arg1 range 711:715 a stable pH range 699:715 a stable pH range of 5.0-11.0 699:727 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 4 8 theme kDa 364:366 arg1 mass 351:354 a calculated molecular mass 328:354 a calculated molecular mass of 82.6 kDa 328:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 5 9 theme GH36 403:406 arg1 organization 408:419 the typical GH36 organization 391:419 the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet 391:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 8 10 theme temperature 757:767 arg1 optimum 769:775 a temperature optimum 755:775 a temperature optimum of 60 °C 755:784 The enzyme had a temperature optimum of 60 °C. 25064132 2 11 theme glycosyl 161:168 arg1 family 180:185 glycosyl hydrolase family 36 161:188 glycosyl hydrolase family 36 161:188 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 5 12 theme antiparallel 513:524 arg1 β-sheet 526:532 the C-terminal antiparallel β-sheet 498:532 the C-terminal antiparallel β-sheet 498:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 5 13 theme catalytic 468:476 arg1 8-barrels 483:491 the catalytic (β/α)8-barrels 464:491 the catalytic (β/α)8-barrels 464:491 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 0 14 from sp 62:63 arg1 α-galactosidase 6:20 A new α-galactosidase 0:20 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp 0:63 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp. 25064132 11 15 theme industrial 1223:1232 arg1 fields 1234:1239 industrial fields 1223:1239 industrial fields 1223:1239 Thus, Gal36A4 may find diverse applications in industrial fields, especially in the food industry. 25064132 8 16 contain had 751:753 arg2 optimum 769:775 a temperature optimum 755:775 a temperature optimum of 60 °C 755:784 The enzyme had a temperature optimum of 60 °C. 25064132 8 16 contain had 751:753 arg1 enzyme 744:749 The enzyme 740:749 The enzyme 740:749 The enzyme had a temperature optimum of 60 °C. 25064132 1 17 theme wide 74:77 arg1 specificity 88:98 wide acceptor specificity 74:98 wide acceptor specificity for transglycosylation 74:121 A4 with wide acceptor specificity for transglycosylation. 25064132 4 18 theme bp 275:276 arg1 ORF 262:264 an ORF 259:264 an ORF of 2,187 bp 259:276 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 19 with polypeptide 292:302 arg1 mass 351:354 a calculated molecular mass 328:354 a calculated molecular mass of 82.6 kDa 328:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 20 contain contains 250:257 arg1 polypeptide 292:302 a polypeptide 290:302 a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa 290:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 20 contain contains 250:257 arg1 It 247:248 It 247:248 It 247:248 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 20 contain contains 250:257 arg2 ORF 262:264 an ORF 259:264 an ORF of 2,187 bp 259:276 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 1 21 theme acceptor 79:86 arg1 specificity 88:98 wide acceptor specificity 74:98 wide acceptor specificity for transglycosylation 74:121 A4 with wide acceptor specificity for transglycosylation. 25064132 5 22 theme β/α 479:481 arg1 8-barrels 483:491 the catalytic (β/α)8-barrels 464:491 the catalytic (β/α)8-barrels 464:491 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 0 23 theme new 2:4 arg1 α-galactosidase 6:20 A new α-galactosidase 0:20 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp 0:63 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp. 25064132 10 24 theme acceptor 1010:1017 arg1 specificity 1019:1029 acceptor specificity 1010:1029 acceptor specificity 1010:1029 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 4 25 theme molecular 341:349 arg1 mass 351:354 a calculated molecular mass 328:354 a calculated molecular mass of 82.6 kDa 328:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 10 26 theme % 983:983 arg1 ratio 968:972 the transfer ratio 955:972 the transfer ratio of 23.25 % 955:983 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 11 27 from applications 1207:1218 arg1 fields 1234:1239 industrial fields 1223:1239 industrial fields 1223:1239 Thus, Gal36A4 may find diverse applications in industrial fields, especially in the food industry. 25064132 5 28 theme N-terminal 443:452 arg1 β-sheets 454:461 three domains--the N-terminal β-sheets 424:461 three domains--the N-terminal β-sheets 424:461 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 5 29 theme β-sheets 454:461 arg1 organization 408:419 the typical GH36 organization 391:419 the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet 391:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 5 30 theme β-sheet 526:532 arg1 organization 408:419 the typical GH36 organization 391:419 the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet 391:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 0 31 theme Alicyclobacillus 45:60 arg1 sp 62:63 thermoacidophilic Alicyclobacillus sp 27:63 thermoacidophilic Alicyclobacillus sp 27:63 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp. 25064132 8 32 theme °C 783:784 arg1 optimum 769:775 a temperature optimum 755:775 a temperature optimum of 60 °C 755:784 The enzyme had a temperature optimum of 60 °C. 25064132 0 33 theme thermoacidophilic 27:43 arg1 sp 62:63 thermoacidophilic Alicyclobacillus sp 27:63 thermoacidophilic Alicyclobacillus sp 27:63 A new α-galactosidase from thermoacidophilic Alicyclobacillus sp. 25064132 7 34 theme stable 701:706 arg1 range 711:715 a stable pH range 699:715 a stable pH range of 5.0-11.0 699:727 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 2 35 theme family 180:185 arg1 gene 143:146 An α-galactosidase gene 124:146 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 124:188 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 2 35 theme family 180:185 arg1 gal36A4 149:155 gal36A4 149:155 gal36A4 149:155 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 2 36 theme sp 239:240 arg1 genome 212:217 the genome 208:217 the genome of Alicyclobacillus sp 208:240 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 5 37 theme C-terminal 502:511 arg1 β-sheet 526:532 the C-terminal antiparallel β-sheet 498:532 the C-terminal antiparallel β-sheet 498:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 1 38 with A4 66:67 arg1 specificity 88:98 wide acceptor specificity 74:98 wide acceptor specificity for transglycosylation 74:121 A4 with wide acceptor specificity for transglycosylation. 25064132 2 39 theme hydrolase 170:178 arg1 family 180:185 glycosyl hydrolase family 36 161:188 glycosyl hydrolase family 36 161:188 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 2 40 theme Alicyclobacillus 222:237 arg1 sp 239:240 Alicyclobacillus sp 222:240 Alicyclobacillus sp 222:240 An α-galactosidase gene (gal36A4) of glycosyl hydrolase family 36 was identified in the genome of Alicyclobacillus sp. 25064132 4 41 theme acids 317:321 arg1 polypeptide 292:302 a polypeptide 290:302 a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa 290:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 41 theme acids 317:321 arg1 It 247:248 It 247:248 It 247:248 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 4 42 theme calculated 330:339 arg1 mass 351:354 a calculated molecular mass 328:354 a calculated molecular mass of 82.6 kDa 328:366 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 25064132 7 43 theme hydrolysis 666:675 arg1 activity 677:684 hydrolysis activity 666:684 hydrolysis activity 666:684 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 10 44 theme galactose 929:937 arg1 acceptor 917:924 acceptor 917:924 acceptor 917:924 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 10 44 theme galactose 929:937 arg1 donor 907:911 donor 907:911 donor 907:911 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 10 45 theme tested 1036:1041 arg1 monosaccharides 1043:1057 all tested monosaccharides 1032:1057 all tested monosaccharides 1032:1057 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 9 46 theme K 842:842 arg1 value 846:850 a K m value 840:850 a K m value of 1.45 mM 840:861 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 9 47 theme m 844:844 arg1 value 846:850 a K m value 840:850 a K m value of 1.45 mM 840:861 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 10 48 theme transfer 959:966 arg1 ratio 968:972 the transfer ratio 955:972 the transfer ratio of 23.25 % 955:983 When using melibiose as both donor and acceptor of galactose, Gal36A4 showed the transfer ratio of 23.25 % at 96 h. With respect to acceptor specificity, all tested monosaccharides, disaccharides, and oligosaccharides except for D-xylose and L-arabinose were good acceptors for transglycosylation. 25064132 5 49 theme 8-barrels 483:491 arg1 organization 408:419 the typical GH36 organization 391:419 the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet 391:532 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 6 50 theme transglycosylation 616:633 arg1 activities 635:644 transglycosylation activities 616:644 transglycosylation activities 616:644 The gene product was produced in Escherichia coli and showed both hydrolysis and transglycosylation activities. 25064132 7 51 theme 5.0-11.0 720:727 arg1 range 711:715 a stable pH range 699:715 a stable pH range of 5.0-11.0 699:727 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 7 52 theme optimal 651:657 arg1 pH 659:660 The optimal pH 647:660 The optimal pH for hydrolysis activity 647:684 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 7 52 theme optimal 651:657 arg1 6.0 690:692 6.0 690:692 6.0 690:692 The optimal pH for hydrolysis activity was 6.0, and a stable pH range of 5.0-11.0 was found. 25064132 9 53 theme α-1,6-glycosidic 806:821 arg1 linkages 823:830 α-1,6-glycosidic linkages 806:830 α-1,6-glycosidic linkages 806:830 It is specific for α-1,6-glycosidic linkages and had a K m value of 1.45 mM toward pNPGal. 25064132 5 54 theme domains--the 430:441 arg1 β-sheets 454:461 three domains--the N-terminal β-sheets 424:461 three domains--the N-terminal β-sheets 424:461 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 5 55 theme Deduced 369:375 arg1 Gal36A4 377:383 Deduced Gal36A4 369:383 Deduced Gal36A4 369:383 Deduced Gal36A4 shows the typical GH36 organization of three domains--the N-terminal β-sheets, the catalytic (β/α)8-barrels, and the C-terminal antiparallel β-sheet. 25064132 6 56 theme gene 539:542 arg1 product 544:550 The gene product 535:550 The gene product 535:550 The gene product was produced in Escherichia coli and showed both hydrolysis and transglycosylation activities. 25064132 4 57 theme amino 311:315 arg1 acids 317:321 728 amino acids 307:321 728 amino acids 307:321 It contains an ORF of 2,187 bp and encodes a polypeptide of 728 amino acids with a calculated molecular mass of 82.6 kDa. 28947169 7 0 theme mannose 1169:1175 arg1 enrichment 1150:1159 enrichment 1150:1159 enrichment of high mannose 1150:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 7 0 theme mannose 1169:1175 arg1 depletion 1137:1145 depletion 1137:1145 depletion 1137:1145 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 8 1 theme normal-phase 1290:1301 arg1 glycans 1313:1319 normal-phase separated glycans 1290:1319 normal-phase separated glycans 1290:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 9 2 theme SPR 1469:1471 arg1 analysis 1488:1495 SPR lectin binding analysis 1469:1495 SPR lectin binding analysis 1469:1495 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 2 theme SPR 1469:1471 arg1 method 1524:1529 a quick alternative method 1504:1529 a quick alternative method to profile protein glycosylation 1504:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 8 3 theme glycan 1271:1276 arg1 analysis 1278:1285 2-AB labeled glycan analysis 1258:1285 2-AB labeled glycan analysis of normal-phase separated glycans 1258:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 9 4 theme lectin 1473:1478 arg1 analysis 1488:1495 SPR lectin binding analysis 1469:1495 SPR lectin binding analysis 1469:1495 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 4 theme lectin 1473:1478 arg1 method 1524:1529 a quick alternative method 1504:1529 a quick alternative method to profile protein glycosylation 1504:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 2 5 from transferrin 309:319 arg1 removal 272:278 sequential removal 261:278 sequential removal of sugar moieties from human transferrin using specific glycosidases 261:347 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 8 6 theme glycan 1424:1429 arg1 profiles 1431:1438 their glycan profiles 1418:1438 their glycan profiles 1418:1438 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 4 7 theme antibodies 575:584 arg1 profiles 535:542 Fc glycosylation profiles 518:542 Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells) 518:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 8 8 theme engineered 1375:1384 arg1 antibodies 1386:1395 the glycan engineered antibodies 1364:1395 the glycan engineered antibodies 1364:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 7 9 with variants 1123:1130 arg1 enrichment 1150:1159 enrichment 1150:1159 enrichment of high mannose 1150:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 7 9 with variants 1123:1130 arg1 depletion 1137:1145 depletion 1137:1145 depletion 1137:1145 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 8 10 theme glycans 1313:1319 arg1 analysis 1278:1285 2-AB labeled glycan analysis 1258:1285 2-AB labeled glycan analysis of normal-phase separated glycans 1258:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 8 11 theme receptor 1335:1342 arg1 activity 1352:1359 Fc gamma receptor binding activity 1326:1359 Fc gamma receptor binding activity of the glycan engineered antibodies 1326:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 4 12 theme mammalian 599:607 arg1 cells 635:639 CHO and HEK 293 6E cells 616:639 cells 635:639 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 12 theme mammalian 599:607 arg1 cells 609:613 mammalian cells 599:613 mammalian cells (CHO and HEK 293 6E cells) 599:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 7 13 theme high 1164:1167 arg1 mannose 1169:1175 high mannose 1164:1175 high mannose 1164:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 6 14 theme expressed 860:868 arg1 antibodies 870:879 CHO cell expressed antibodies 851:879 CHO cell expressed antibodies 851:879 CHO cell expressed antibodies were also more heavily fucosylated than the ones expressed by 293 6E cells. 28947169 8 15 theme separated 1303:1311 arg1 glycans 1313:1319 normal-phase separated glycans 1290:1319 normal-phase separated glycans 1290:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 5 16 contain contained 828:836 arg2 acid 845:848 sialic acid 838:848 sialic acid 838:848 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 5 16 contain contained 828:836 arg1 antibodies 791:800 antibodies 791:800 antibodies expressed in 293 6E cells 791:826 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 2 17 theme sequential 261:270 arg1 removal 272:278 sequential removal 261:278 sequential removal of sugar moieties from human transferrin using specific glycosidases 261:347 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 6 18 theme 6E 947:948 arg1 cells 950:954 293 6E cells 943:954 293 6E cells 943:954 CHO cell expressed antibodies were also more heavily fucosylated than the ones expressed by 293 6E cells. 28947169 6 19 gly fucosylated 904:914 arg1 antibodies 870:879 CHO cell expressed antibodies 851:879 CHO cell expressed antibodies 851:879 CHO cell expressed antibodies were also more heavily fucosylated than the ones expressed by 293 6E cells. 28947169 9 20 theme alternative 1512:1522 arg1 analysis 1488:1495 SPR lectin binding analysis 1469:1495 SPR lectin binding analysis 1469:1495 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 20 theme alternative 1512:1522 arg1 method 1524:1529 a quick alternative method 1504:1529 a quick alternative method to profile protein glycosylation 1504:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 5 21 theme sialic 838:843 arg1 acid 845:848 sialic acid 838:848 sialic acid 838:848 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 1 22 theme Surface 162:168 arg1 Resonance 178:186 Surface Plasmon Resonance 162:186 Surface Plasmon Resonance (SPR) 162:192 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 1 22 theme Surface 162:168 arg1 SPR 189:191 SPR 189:191 SPR 189:191 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 6 23 theme CHO 851:853 arg1 cell 855:858 CHO cell 851:858 CHO cell 851:858 CHO cell expressed antibodies were also more heavily fucosylated than the ones expressed by 293 6E cells. 28947169 1 24 theme Plasmon 170:176 arg1 Resonance 178:186 Surface Plasmon Resonance 162:186 Surface Plasmon Resonance (SPR) 162:192 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 1 24 theme Plasmon 170:176 arg1 SPR 189:191 SPR 189:191 SPR 189:191 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 3 25 from glycans 385:391 arg1 protein 400:406 the protein 396:406 the protein 396:406 The results clearly indicated that glycans on the protein can be identified by their selective binding activity to various lectins. 28947169 7 26 theme mouse 1043:1047 arg1 antibodies 1049:1058 glycan engineered mouse antibodies 1025:1058 glycan engineered mouse antibodies 1025:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 1 27 theme Glycan 80:85 arg1 profiling 87:95 Glycan profiling 80:95 Glycan profiling of proteins 80:107 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 4 28 theme E. coli 648:654 arg1 proteins 666:673 E. coli expressed proteins 648:673 E. coli expressed proteins as the negative controls 648:698 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 0 29 theme Glycan 0:5 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling of proteins 0:27 Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance. 28947169 4 30 theme therapeutic 547:557 arg1 peptibodies 559:569 therapeutic peptibodies 547:569 therapeutic peptibodies 547:569 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 7 31 theme antibodies 1049:1058 arg1 composition 1010:1020 the fucose composition 999:1020 the fucose composition of glycan engineered mouse antibodies 999:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 8 32 from analysis 1278:1285 arg1 results 1245:1251 results 1245:1251 results from 2-AB labeled glycan analysis of normal-phase separated glycans 1245:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 2 33 theme moieties 289:296 arg1 removal 272:278 sequential removal 261:278 sequential removal of sugar moieties from human transferrin using specific glycosidases 261:347 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 9 34 theme profile 1534:1540 arg1 glycosylation 1550:1562 profile protein glycosylation 1534:1562 profile protein glycosylation 1534:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 35 theme quick 1506:1510 arg1 analysis 1488:1495 SPR lectin binding analysis 1469:1495 SPR lectin binding analysis 1469:1495 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 35 theme quick 1506:1510 arg1 method 1524:1529 a quick alternative method 1504:1529 a quick alternative method to profile protein glycosylation 1504:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 0 36 theme proteins 20:27 arg1 profiling 7:15 Glycan profiling 0:15 Glycan profiling of proteins 0:27 Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance. 28947169 7 37 theme glycan 1025:1030 arg1 antibodies 1049:1058 glycan engineered mouse antibodies 1025:1058 glycan engineered mouse antibodies 1025:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 2 38 theme sugar 283:287 arg1 moieties 289:296 sugar moieties 283:296 sugar moieties 283:296 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 7 39 theme variants 1123:1130 arg1 composition 1093:1103 mannose composition 1085:1103 mannose composition of human antibody variants with depletion or enrichment of high mannose 1085:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 7 40 theme engineered 1032:1041 arg1 antibodies 1049:1058 glycan engineered mouse antibodies 1025:1058 glycan engineered mouse antibodies 1025:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 1 41 theme proteins 100:107 arg1 profiling 87:95 Glycan profiling 80:95 Glycan profiling of proteins 80:107 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 4 42 theme HEK 624:626 arg1 cells 635:639 CHO and HEK 293 6E cells 616:639 cells 635:639 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 42 theme HEK 624:626 arg1 cells 609:613 mammalian cells 599:613 mammalian cells (CHO and HEK 293 6E cells) 599:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 0 43 theme lectin 35:40 arg1 binding 42:48 lectin binding 35:48 lectin binding by Surface Plasmon Resonance 35:77 Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance. 28947169 4 44 theme peptibodies 559:569 arg1 profiles 535:542 Fc glycosylation profiles 518:542 Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells) 518:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 7 45 dep applied 968:974 arg1 measure 991:997 measure 991:997 to measure the fucose composition of glycan engineered mouse antibodies 988:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 7 45 dep applied 968:974 arg1 determine 1075:1083 determine 1075:1083 to determine mannose composition of human antibody variants with depletion or enrichment of high mannose 1072:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 3 46 theme selective 435:443 arg1 activity 453:460 their selective binding activity 429:460 their selective binding activity to various lectins 429:479 The results clearly indicated that glycans on the protein can be identified by their selective binding activity to various lectins. 28947169 7 47 theme human 1108:1112 arg1 variants 1123:1130 human antibody variants 1108:1130 human antibody variants with depletion or enrichment of high mannose 1108:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 4 48 theme Fc 518:519 arg1 profiles 535:542 Fc glycosylation profiles 518:542 Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells) 518:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 7 49 theme antibody 1114:1121 arg1 variants 1123:1130 human antibody variants 1108:1130 human antibody variants with depletion or enrichment of high mannose 1108:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 5 50 theme 6E 819:820 arg1 cells 822:826 293 6E cells 815:826 293 6E cells 815:826 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 3 51 theme binding 445:451 arg1 activity 453:460 their selective binding activity 429:460 their selective binding activity to various lectins 429:479 The results clearly indicated that glycans on the protein can be identified by their selective binding activity to various lectins. 28947169 7 52 theme mannose 1085:1091 arg1 composition 1093:1103 mannose composition 1085:1103 mannose composition of human antibody variants with depletion or enrichment of high mannose 1085:1175 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 8 53 with consistent 1402:1411 arg1 profiles 1431:1438 their glycan profiles 1418:1438 their glycan profiles 1418:1438 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 8 54 theme gamma 1329:1333 arg1 receptor 1335:1342 Fc gamma receptor 1326:1342 Fc gamma receptor binding activity of the glycan engineered antibodies 1326:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 5 55 theme CHO 742:744 arg1 cells 746:750 CHO cells 742:750 CHO cells 742:750 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 8 56 theme glycan 1368:1373 arg1 antibodies 1386:1395 the glycan engineered antibodies 1364:1395 the glycan engineered antibodies 1364:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 8 57 theme Fc 1326:1327 arg1 receptor 1335:1342 Fc gamma receptor 1326:1342 Fc gamma receptor binding activity of the glycan engineered antibodies 1326:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 2 58 theme specific 327:334 arg1 glycosidases 336:347 specific glycosidases 327:347 specific glycosidases 327:347 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 5 59 theme sialic 772:777 arg1 acid 779:782 any sialic acid 768:782 any sialic acid 768:782 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 7 60 theme fucose 1003:1008 arg1 composition 1010:1020 the fucose composition 999:1020 the fucose composition of glycan engineered mouse antibodies 999:1058 We further applied this method to measure the fucose composition of glycan engineered mouse antibodies, as well as to determine mannose composition of human antibody variants with depletion or enrichment of high mannose. 28947169 4 61 theme negative 682:689 arg1 controls 691:698 the negative controls 678:698 the negative controls 678:698 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 62 theme glycosylation 521:533 arg1 profiles 535:542 Fc glycosylation profiles 518:542 Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells) 518:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 8 63 theme antibodies 1386:1395 arg1 activity 1352:1359 Fc gamma receptor binding activity 1326:1359 Fc gamma receptor binding activity of the glycan engineered antibodies 1326:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 2 64 theme lectin 241:246 arg1 binding 248:254 specific lectin binding 232:254 specific lectin binding 232:254 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 4 65 theme expressed 656:664 arg1 proteins 666:673 E. coli expressed proteins 648:673 E. coli expressed proteins as the negative controls 648:698 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 3 66 theme various 465:471 arg1 lectins 473:479 various lectins 465:479 various lectins 465:479 The results clearly indicated that glycans on the protein can be identified by their selective binding activity to various lectins. 28947169 8 67 theme binding 1344:1350 arg1 activity 1352:1359 Fc gamma receptor binding activity 1326:1359 Fc gamma receptor binding activity of the glycan engineered antibodies 1326:1395 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 5 68 contain contain 760:766 arg1 antibodies 718:727 antibodies 718:727 antibodies expressed in CHO cells 718:750 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 5 68 contain contain 760:766 arg2 acid 779:782 any sialic acid 768:782 any sialic acid 768:782 We observed that antibodies expressed in CHO cells did not contain any sialic acid, while antibodies expressed in 293 6E cells contained sialic acid. 28947169 2 69 theme specific 232:239 arg1 binding 248:254 specific lectin binding 232:254 specific lectin binding 232:254 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 2 70 theme human 303:307 arg1 transferrin 309:319 human transferrin 303:319 human transferrin using specific glycosidases 303:347 To validate the method, we monitored specific lectin binding with sequential removal of sugar moieties from human transferrin using specific glycosidases. 28947169 8 71 theme glycan 1182:1187 arg1 profiles 1189:1196 The glycan profiles 1178:1196 The glycan profiles generated using this method 1178:1224 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 8 71 theme glycan 1182:1187 arg1 comparable 1231:1240 comparable 1231:1240 comparable 1231:1240 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 8 72 theme labeled 1263:1269 arg1 analysis 1278:1285 2-AB labeled glycan analysis 1258:1285 2-AB labeled glycan analysis of normal-phase separated glycans 1258:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 0 73 theme Plasmon 61:67 arg1 Resonance 69:77 Surface Plasmon Resonance 53:77 Surface Plasmon Resonance 53:77 Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance. 28947169 4 74 theme 6E 632:633 arg1 cells 635:639 CHO and HEK 293 6E cells 616:639 cells 635:639 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 74 theme 6E 632:633 arg1 cells 609:613 mammalian cells 599:613 mammalian cells (CHO and HEK 293 6E cells) 599:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 9 75 theme binding 1480:1486 arg1 analysis 1488:1495 SPR lectin binding analysis 1469:1495 SPR lectin binding analysis 1469:1495 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 9 75 theme binding 1480:1486 arg1 method 1524:1529 a quick alternative method 1504:1529 a quick alternative method to profile protein glycosylation 1504:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 1 76 theme lectin 135:140 arg1 activity 150:157 their lectin binding activity 129:157 their lectin binding activity 129:157 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 8 77 theme 2-AB 1258:1261 arg1 analysis 1278:1285 2-AB labeled glycan analysis 1258:1285 2-AB labeled glycan analysis of normal-phase separated glycans 1258:1319 The glycan profiles generated using this method were comparable to results from 2-AB labeled glycan analysis of normal-phase separated glycans, and Fc gamma receptor binding activity of the glycan engineered antibodies were consistent with their glycan profiles. 28947169 6 78 dep expressed 860:868 arg1 cell 855:858 CHO cell 851:858 CHO cell 851:858 CHO cell expressed antibodies were also more heavily fucosylated than the ones expressed by 293 6E cells. 28947169 0 79 theme Surface 53:59 arg1 Resonance 69:77 Surface Plasmon Resonance 53:77 Surface Plasmon Resonance 53:77 Glycan profiling of proteins using lectin binding by Surface Plasmon Resonance. 28947169 9 80 theme protein 1542:1548 arg1 glycosylation 1550:1562 profile protein glycosylation 1534:1562 profile protein glycosylation 1534:1562 Hence, we demonstrated that SPR lectin binding analysis can be a quick alternative method to profile protein glycosylation. 28947169 4 81 gly glycosylation 521:533 arg1 antibodies 575:584 antibodies 575:584 antibodies 575:584 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 81 gly glycosylation 521:533 arg1 peptibodies 559:569 therapeutic peptibodies 547:569 therapeutic peptibodies 547:569 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 1 82 theme binding 142:148 arg1 activity 150:157 their lectin binding activity 129:157 their lectin binding activity 129:157 Glycan profiling of proteins was studied through their lectin binding activity by Surface Plasmon Resonance (SPR). 28947169 4 83 theme CHO 616:618 arg1 cells 635:639 CHO and HEK 293 6E cells 616:639 cells 635:639 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 28947169 4 83 theme CHO 616:618 arg1 cells 609:613 mammalian cells 599:613 mammalian cells (CHO and HEK 293 6E cells) 599:640 Using this method, we characterized Fc glycosylation profiles of therapeutic peptibodies and antibodies expressed in mammalian cells (CHO and HEK 293 6E cells), with E. coli expressed proteins as the negative controls. 24326352 4 0 theme residues 701:708 arg1 residues 701:708 specific sugar residues 686:708 specific sugar residues 686:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 0 theme residues 701:708 arg1 amounts 675:681 the total amounts 665:681 the total amounts of specific sugar residues 665:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 5 1 theme large 1020:1024 arg1 components 1037:1046 the dimeric and large aggregated components 1004:1046 the dimeric and large aggregated components 1004:1046 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 0 2 theme ovary 77:81 arg1 cells 83:87 Chinese hamster ovary cells 61:87 Chinese hamster ovary cells in bioreactor culture 61:109 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 4 3 from monomers 844:851 arg1 % 825:825 70-80% 820:825 70-80% of the amount in monomers 820:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 3 from monomers 844:851 arg1 amount 834:839 the amount 830:839 the amount in monomers 830:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 6 4 theme dimers 1272:1277 arg1 formation 1259:1267 the formation 1255:1267 the formation of dimers 1255:1277 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 3 5 theme glycosylation 526:538 arg1 patterns 540:547 the glycosylation patterns 522:547 the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody 522:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 0 6 theme bioreactor 92:101 arg1 culture 103:109 bioreactor culture 92:109 bioreactor culture 92:109 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 6 7 theme cells 1202:1206 arg1 aggregation 1149:1159 antibody aggregation 1140:1159 antibody aggregation in cell culture of Chinese hamster ovary cells 1140:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 4 8 from residues 764:771 arg1 dimers 776:781 dimers 776:781 dimers 776:781 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 8 from residues 764:771 arg1 aggregates 793:802 large aggregates 787:802 large aggregates 787:802 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 3 9 theme dimeric 563:569 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 3 10 theme aggregated 582:591 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 4 11 from amount 834:839 arg1 monomers 844:851 monomers 844:851 monomers 844:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 5 12 theme deglycosylation 941:955 arg1 oligosaccharides 911:926 N-linked oligosaccharides 902:926 N-linked oligosaccharides 902:926 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 12 theme deglycosylation 941:955 arg1 result 931:936 a result 929:936 a result of deglycosylation or aglycosylation 929:973 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 6 13 from culture 1169:1175 arg1 heterogeneities 1099:1113 glycosylation heterogeneities 1085:1113 glycosylation heterogeneities 1085:1113 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 6 13 from culture 1169:1175 arg1 cause 1131:1135 a potential cause 1119:1135 a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells 1119:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 3 14 theme apparent 499:506 arg1 difference 508:517 no apparent difference 496:517 no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody 496:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 4 15 theme sugar 695:699 arg1 residues 701:708 specific sugar residues 686:708 specific sugar residues 686:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 16 theme binding 636:642 arg1 assays 644:649 lectin binding assays 629:649 lectin binding assays 629:649 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 3 17 gly glycosylation 526:538 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 2 18 theme Chinese 397:403 arg1 cells 419:423 Chinese hamster ovary cells 397:423 Chinese hamster ovary cells 397:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 5 19 theme oligosaccharides 911:926 arg1 lack 894:897 the lack 890:897 the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, 890:974 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 1 20 theme biological 184:193 arg1 function 195:202 their biological function 178:202 their biological function 178:202 N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. 24326352 4 21 from % 825:825 arg1 monomers 844:851 monomers 844:851 monomers 844:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 22 theme amount 834:839 arg1 % 825:825 70-80% 820:825 70-80% of the amount in monomers 820:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 22 theme amount 834:839 arg1 amount 834:839 the amount 830:839 the amount in monomers 830:851 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 2 23 gly glycosylation 300:312 arg1 antibody 338:345 an aggregated antibody 324:345 an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells 324:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 2 24 theme status 314:319 arg1 impact 286:291 the impact 282:291 the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells 282:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 3 25 theme monomeric 552:560 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 3 26 theme High-performance 426:441 arg1 chromatography 450:463 High-performance liquid chromatography 426:463 High-performance liquid chromatography analysis 426:472 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 0 27 theme Glycosylation 0:12 arg1 analysis 14:21 Glycosylation analysis 0:21 Glycosylation analysis of an aggregated antibody 0:47 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 3 28 from difference 508:517 arg1 patterns 540:547 the glycosylation patterns 522:547 the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody 522:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 2 29 theme glycosylation 300:312 arg1 status 314:319 the glycosylation status 296:319 the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells 296:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 2 30 theme bioreactor 375:384 arg1 culture 386:392 the bioreactor culture 371:392 the bioreactor culture of Chinese hamster ovary cells 371:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 5 31 theme N-linked 902:909 arg1 oligosaccharides 911:926 N-linked oligosaccharides 902:926 N-linked oligosaccharides 902:926 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 31 theme N-linked 902:909 arg1 result 931:936 a result 929:936 a result of deglycosylation or aglycosylation 929:973 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 4 32 theme galactose 743:751 arg1 residues 764:771 both galactose and fucose residues 738:771 residues 764:771 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 3 33 theme forms 593:597 arg1 patterns 540:547 the glycosylation patterns 522:547 the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody 522:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 6 34 theme Chinese 1180:1186 arg1 cells 1202:1206 Chinese hamster ovary cells 1180:1206 Chinese hamster ovary cells 1180:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 1 35 gly N-Glycosylation 112:126 arg1 antibodies 143:152 therapeutic antibodies 131:152 therapeutic antibodies 131:152 N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. 24326352 0 36 from cells 83:87 arg1 culture 103:109 bioreactor culture 92:109 bioreactor culture 92:109 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 3 37 theme large 576:580 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 6 38 from aggregation 1149:1159 arg1 culture 1169:1175 cell culture 1164:1175 cell culture 1164:1175 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 2 39 theme cells 419:423 arg1 culture 386:392 the bioreactor culture 371:392 the bioreactor culture of Chinese hamster ovary cells 371:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 0 40 theme antibody 40:47 arg1 analysis 14:21 Glycosylation analysis 0:21 Glycosylation analysis of an aggregated antibody 0:47 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 6 41 theme potential 1121:1129 arg1 heterogeneities 1099:1113 glycosylation heterogeneities 1085:1113 glycosylation heterogeneities 1085:1113 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 6 41 theme potential 1121:1129 arg1 cause 1131:1135 a potential cause 1119:1135 a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells 1119:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 3 42 theme antibody 606:613 arg1 forms 593:597 monomeric, dimeric, and large aggregated forms 552:597 monomeric, dimeric, and large aggregated forms of the antibody 552:613 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 3 43 theme liquid 443:448 arg1 chromatography 450:463 High-performance liquid chromatography 426:463 High-performance liquid chromatography analysis 426:472 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 2 44 theme ovary 413:417 arg1 cells 419:423 Chinese hamster ovary cells 397:423 Chinese hamster ovary cells 397:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 0 45 theme aggregated 29:38 arg1 antibody 40:47 an aggregated antibody 26:47 an aggregated antibody 26:47 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 6 46 theme aggregation 1149:1159 arg1 heterogeneities 1099:1113 glycosylation heterogeneities 1085:1113 glycosylation heterogeneities 1085:1113 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 6 46 theme aggregation 1149:1159 arg1 cause 1131:1135 a potential cause 1119:1135 a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells 1119:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 2 47 theme hamster 405:411 arg1 cells 419:423 Chinese hamster ovary cells 397:423 Chinese hamster ovary cells 397:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 6 48 theme large 1302:1306 arg1 aggregates 1308:1317 large aggregates 1302:1317 large aggregates 1302:1317 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 3 49 theme chromatography 450:463 arg1 analysis 465:472 High-performance liquid chromatography analysis 426:472 High-performance liquid chromatography analysis 426:472 High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. 24326352 2 50 theme antibody 338:345 arg1 status 314:319 the glycosylation status 296:319 the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells 296:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 5 51 link N-linked 902:909 arg1 oligosaccharides 911:926 N-linked oligosaccharides 902:926 N-linked oligosaccharides 902:926 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 51 link N-linked 902:909 arg1 result 931:936 a result 929:936 a result of deglycosylation or aglycosylation 929:973 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 52 theme dimeric 1008:1014 arg1 components 1037:1046 the dimeric and large aggregated components 1004:1046 the dimeric and large aggregated components 1004:1046 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 6 53 from cause 1131:1135 arg1 culture 1169:1175 cell culture 1164:1175 cell culture 1164:1175 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 2 54 theme aggregated 327:336 arg1 antibody 338:345 an aggregated antibody 324:345 an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells 324:423 Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. 24326352 6 55 theme hamster 1188:1194 arg1 cells 1202:1206 Chinese hamster ovary cells 1180:1206 Chinese hamster ovary cells 1180:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 4 56 theme total 669:673 arg1 residues 701:708 specific sugar residues 686:708 specific sugar residues 686:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 4 56 theme total 669:673 arg1 amounts 675:681 the total amounts 665:681 the total amounts of specific sugar residues 665:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 6 57 theme N-glycosylation 1230:1244 arg1 lack 1222:1225 the lack 1218:1225 the lack of N-glycosylation 1218:1244 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 1 58 theme therapeutic 131:141 arg1 antibodies 143:152 therapeutic antibodies 131:152 therapeutic antibodies 131:152 N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. 24326352 6 59 theme antibody 1140:1147 arg1 aggregation 1149:1159 antibody aggregation 1140:1159 antibody aggregation in cell culture of Chinese hamster ovary cells 1140:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 5 60 theme aglycosylation 960:973 arg1 oligosaccharides 911:926 N-linked oligosaccharides 902:926 N-linked oligosaccharides 902:926 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 60 theme aglycosylation 960:973 arg1 result 931:936 a result 929:936 a result of deglycosylation or aglycosylation 929:973 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 5 61 theme aggregated 1026:1035 arg1 components 1037:1046 the dimeric and large aggregated components 1004:1046 the dimeric and large aggregated components 1004:1046 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 4 62 theme fucose 757:762 arg1 residues 764:771 both galactose and fucose residues 738:771 residues 764:771 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 1 63 theme antibodies 143:152 arg1 N-Glycosylation 112:126 N-Glycosylation 112:126 N-Glycosylation of therapeutic antibodies 112:152 N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. 24326352 0 64 theme hamster 69:75 arg1 cells 83:87 Chinese hamster ovary cells 61:87 Chinese hamster ovary cells in bioreactor culture 61:109 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 5 65 theme components 1037:1046 arg1 proportion 990:999 a proportion 988:999 a proportion of the dimeric and large aggregated components 988:1046 These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. 24326352 6 66 theme ovary 1196:1200 arg1 cells 1202:1206 Chinese hamster ovary cells 1180:1206 Chinese hamster ovary cells 1180:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 6 67 theme cell 1164:1167 arg1 culture 1169:1175 cell culture 1164:1175 cell culture 1164:1175 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 4 68 theme specific 686:693 arg1 residues 701:708 specific sugar residues 686:708 specific sugar residues 686:708 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 6 69 theme present 1053:1059 arg1 study 1061:1065 The present study 1049:1065 The present study 1049:1065 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 4 70 theme lectin 629:634 arg1 assays 644:649 lectin binding assays 629:649 lectin binding assays 629:649 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 0 71 theme Chinese 61:67 arg1 cells 83:87 Chinese hamster ovary cells 61:87 Chinese hamster ovary cells in bioreactor culture 61:109 Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture. 24326352 4 72 theme large 787:791 arg1 aggregates 793:802 large aggregates 787:802 large aggregates 787:802 In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. 24326352 6 73 theme glycosylation 1085:1097 arg1 heterogeneities 1099:1113 glycosylation heterogeneities 1085:1113 glycosylation heterogeneities 1085:1113 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 24326352 6 73 theme glycosylation 1085:1097 arg1 cause 1131:1135 a potential cause 1119:1135 a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells 1119:1206 The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates. 27194760 2 0 with UUKV 334:337 arg1 genome 346:351 a genome 344:351 a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir 344:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 7 1 theme tick 1413:1416 arg1 viruses 1431:1437 tick cell-derived viruses 1413:1437 tick cell-derived viruses 1413:1437 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 3 2 theme many 610:613 arg1 weeks 615:619 many weeks 610:619 many weeks 610:619 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 2 3 theme genetics 308:315 arg1 system 317:322 a reverse genetics system 298:322 a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir 298:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 1 4 from UNLABELLED 45:54 arg1 decade 68:73 the last decade 59:73 the last decade 59:73 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 14 5 theme structural 2434:2443 arg1 properties 2445:2454 specific structural properties 2425:2454 specific structural properties 2425:2454 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 6 6 theme PNGase 1111:1116 arg1 F 1118:1118 PNGase F 1111:1118 PNGase F 1111:1118 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 2 7 theme tick-mammal 248:258 arg1 switch 260:265 the tick-mammal switch 244:265 the tick-mammal switch 244:265 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 14 8 theme tick 2409:2412 arg1 cells 2414:2418 tick cells 2409:2418 tick cells 2409:2418 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 12 9 theme alternation 2123:2133 arg1 importance 2104:2113 the importance 2100:2113 the importance of host alternation in virus transmission to humans and initial infection at the molecular level 2100:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 6 10 theme protein 1161:1167 arg1 mobility 1145:1152 the electrophoretic mobility 1125:1152 the electrophoretic mobility of the protein 1125:1167 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 6 10 theme protein 1161:1167 arg1 slower 1187:1192 slower 1187:1192 slower 1187:1192 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 11 11 theme virus 2024:2028 arg1 stocks 2030:2035 virus stocks 2024:2035 virus stocks produced in mammalian cells 2024:2063 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 12 12 from level 2206:2210 arg1 transmission 2144:2155 virus transmission 2138:2155 virus transmission to humans 2138:2165 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 12 12 from level 2206:2210 arg1 importance 2104:2113 the importance 2100:2113 the importance of host alternation in virus transmission to humans and initial infection at the molecular level 2100:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 12 12 from level 2206:2210 arg1 infection 2179:2187 initial infection 2171:2187 initial infection at the molecular level 2171:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 1 13 theme Uukuniemi 169:177 arg1 UUKV 186:189 UUKV 186:189 UUKV 186:189 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 1 13 theme Uukuniemi 169:177 arg1 virus 179:183 Uukuniemi virus 169:183 Uukuniemi virus (UUKV) 169:190 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 6 14 theme GN 1063:1064 arg1 glycosylations 1066:1079 GN glycosylations 1063:1079 GN glycosylations 1063:1079 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 15 15 theme tick-derived 2568:2579 arg1 particles 2587:2595 tick-derived viral particles 2568:2595 tick-derived viral particles 2568:2595 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 5 16 theme viral 771:775 arg1 protein 777:783 The second envelope viral protein 751:783 The second envelope viral protein 751:783 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 16 theme viral 771:775 arg1 GN 786:787 GN 786:787 GN 786:787 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 13 17 theme in 2251:2252 arg1 model 2276:2280 an in vitro tick cell-based model 2248:2280 an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers 2248:2352 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 6 18 with Treatment 927:935 arg1 β-mercaptoethanol 942:958 β-mercaptoethanol 942:958 β-mercaptoethanol 942:958 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 9 19 theme cell 1762:1765 arg1 biology 1767:1773 the cell biology 1758:1773 the cell biology of arbovirus transmission 1758:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 7 20 theme forming 1291:1297 arg1 unit 1299:1302 focus forming unit 1285:1302 focus forming unit 1285:1302 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 5 21 theme classical 837:845 arg1 F 882:882 peptide-N-glycosidase F 860:882 peptide-N-glycosidase F (PNGase F) 860:893 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 21 theme classical 837:845 arg1 glycosidases 847:858 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 21 theme classical 837:845 arg1 H 915:915 endoglycosidase H 899:915 endoglycosidase H (Endo H) 899:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 9 22 theme working 1676:1682 arg1 entry 1805:1809 entry 1805:1809 entry into mammalian hosts 1805:1830 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 9 22 theme working 1676:1682 arg1 importance 1662:1671 the importance 1658:1671 the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission 1658:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 16 23 theme future 2773:2778 arg1 research 2780:2787 future research 2773:2787 future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks 2773:2886 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 5 24 theme PNGase 885:890 arg1 F 882:882 peptide-N-glycosidase F 860:882 peptide-N-glycosidase F (PNGase F) 860:893 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 24 theme PNGase 885:890 arg1 F 892:892 PNGase F 885:892 PNGase F 885:892 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 6 25 theme disulfide 1217:1225 arg1 bonds 1227:1231 disulfide bonds 1217:1231 disulfide bonds 1217:1231 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 3 26 theme plasmid 554:560 arg1 DNAs 562:565 plasmid DNAs 554:565 plasmid DNAs 554:565 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 5 27 theme peptide-N-glycosidase 860:880 arg1 F 882:882 peptide-N-glycosidase F 860:882 peptide-N-glycosidase F (PNGase F) 860:893 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 27 theme peptide-N-glycosidase 860:880 arg1 F 892:892 PNGase F 885:892 PNGase F 885:892 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 27 theme peptide-N-glycosidase 860:880 arg1 glycosidases 847:858 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 7 28 theme viral 1261:1265 arg1 nucleoprotein 1267:1279 viral nucleoprotein 1261:1279 viral nucleoprotein 1261:1279 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 15 29 theme viral 2581:2585 arg1 particles 2587:2595 tick-derived viral particles 2568:2595 tick-derived viral particles 2568:2595 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 3 30 dep I. 499:500 arg1 ricinus 502:508 I. ricinus 499:508 I. ricinus 499:508 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 4 31 gly mannosylated 702:713 arg1 GC 681:682 The glycoprotein GC 664:682 The glycoprotein GC 664:682 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 9 32 theme vector 1724:1729 arg1 cells 1731:1735 arthropod vector cells 1714:1735 arthropod vector cells in investigations of the cell biology of arbovirus transmission 1714:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 11 33 theme emerging 1967:1974 arg1 diseases 1976:1983 infectious emerging diseases 1956:1983 infectious emerging diseases 1956:1983 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 5 34 contain carried 790:796 arg1 protein 777:783 The second envelope viral protein 751:783 The second envelope viral protein 751:783 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 34 contain carried 790:796 arg2 N-glycans 805:813 N-glycans 805:813 N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 805:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 34 contain carried 790:796 arg1 GN 786:787 GN 786:787 GN 786:787 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 35 dep glycosidases 847:858 arg1 H 923:923 Endo H 918:923 Endo H 918:923 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 35 dep glycosidases 847:858 arg1 F 882:882 peptide-N-glycosidase F 860:882 peptide-N-glycosidase F (PNGase F) 860:893 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 35 dep glycosidases 847:858 arg1 F 892:892 PNGase F 885:892 PNGase F 885:892 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 35 dep glycosidases 847:858 arg1 glycosidases 847:858 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 35 dep glycosidases 847:858 arg1 H 915:915 endoglycosidase H 899:915 endoglycosidase H (Endo H) 899:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 12 36 theme molecular 2196:2204 arg1 level 2206:2210 the molecular level 2192:2210 the molecular level 2192:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 2 37 dep Ixodes 408:413 arg1 ricinus 415:421 ricinus 415:421 ricinus 415:421 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 13 38 theme tick 2260:2263 arg1 model 2276:2280 an in vitro tick cell-based model 2248:2280 an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers 2248:2352 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 15 39 theme receptor/cell 2698:2710 arg1 interactions 2712:2723 early virus-mammalian receptor/cell interactions 2676:2723 early virus-mammalian receptor/cell interactions 2676:2723 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 1 40 theme last 63:66 arg1 decade 68:73 the last decade 59:73 the last decade 59:73 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 3 41 attach derived 486:492 arg1 I. 499:500 I. 499:500 I. 499:500 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 3 41 attach derived 486:492 arg2 IRE/CTVM19 443:452 The IRE/CTVM19 and IRE/CTVM20 cell lines 439:478 IRE/CTVM19 443:452 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 3 41 attach derived 486:492 arg2 both 481:484 both 481:484 both 481:484 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 6 42 theme apparent 979:986 arg1 weight 998:1003 the apparent molecular weight 975:1003 the apparent molecular weight of GN 975:1009 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 2 43 theme authentic 374:382 arg1 virus 384:388 the authentic virus 370:388 the authentic virus isolated from the Ixodes ricinus tick reservoir 370:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 14 44 contain have 2420:2423 arg2 N-glycans 2460:2468 N-glycans 2460:2468 N-glycans 2460:2468 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 14 44 contain have 2420:2423 arg1 virions 2388:2394 virions 2388:2394 virions derived from tick cells 2388:2418 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 14 44 contain have 2420:2423 arg2 properties 2445:2454 specific structural properties 2425:2454 specific structural properties 2425:2454 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 5 45 theme second 755:760 arg1 protein 777:783 The second envelope viral protein 751:783 The second envelope viral protein 751:783 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 45 theme second 755:760 arg1 GN 786:787 GN 786:787 GN 786:787 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 1 46 theme tick-borne 82:91 arg1 phleboviruses 104:116 novel tick-borne pathogenic phleboviruses 76:116 novel tick-borne pathogenic phleboviruses in the family Bunyaviridae 76:143 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 1 46 theme tick-borne 82:91 arg1 all 146:148 all 146:148 all 146:148 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 4 47 link cell-derived 723:734 arg1 progeny 742:748 tick cell-derived viral progeny 718:748 tick cell-derived viral progeny 718:748 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 0 48 theme Tick-Borne 21:30 arg1 Model 38:42 a Tick-Borne Virus Model 19:42 a Tick-Borne Virus Model 19:42 Uukuniemi Virus as a Tick-Borne Virus Model. 27194760 0 48 theme Tick-Borne 21:30 arg1 Virus 10:14 Virus 10:14 Virus 10:14 Uukuniemi Virus as a Tick-Borne Virus Model. 27194760 2 49 theme Ixodes 408:413 arg1 reservoir 428:436 the Ixodes ricinus tick reservoir 404:436 the Ixodes ricinus tick reservoir 404:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 9 50 theme transmission 1788:1799 arg1 biology 1767:1773 the cell biology 1758:1773 the cell biology of arbovirus transmission 1758:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 2 51 theme reverse 300:306 arg1 system 317:322 a reverse genetics system 298:322 a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir 298:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 15 52 theme virus-mammalian 2682:2696 arg1 interactions 2712:2723 early virus-mammalian receptor/cell interactions 2676:2723 early virus-mammalian receptor/cell interactions 2676:2723 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 14 53 theme virus 2487:2491 arg1 infectivity 2493:2503 virus infectivity 2487:2503 virus infectivity 2487:2503 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 6 54 theme BHK-21 1049:1054 arg1 cells 1056:1060 mammalian BHK-21 cells 1039:1060 mammalian BHK-21 cells 1039:1060 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 7 55 link cell-derived 1418:1429 arg1 viruses 1431:1437 tick cell-derived viruses 1413:1437 tick cell-derived viruses 1413:1437 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 8 56 theme tick 1511:1514 arg1 cells 1516:1520 vector tick cells 1504:1520 vector tick cells 1504:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 10 57 theme Tick-borne 1844:1853 arg1 phleboviruses 1855:1867 IMPORTANCE Tick-borne phleboviruses 1833:1867 IMPORTANCE Tick-borne phleboviruses 1833:1867 IMPORTANCE Tick-borne phleboviruses represent a growing threat to humans globally. 27194760 12 58 theme virus 2138:2142 arg1 transmission 2144:2155 virus transmission 2138:2155 virus transmission to humans 2138:2165 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 16 59 theme tick-borne 2801:2810 arg1 phleboviruses 2812:2824 tick-borne phleboviruses 2801:2824 not only tick-borne phleboviruses but also all viruses 2792:2845 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 7 60 theme cell-derived 1418:1429 arg1 viruses 1431:1437 tick cell-derived viruses 1413:1437 tick cell-derived viruses 1413:1437 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 5 61 theme Endo 918:921 arg1 H 923:923 Endo H 918:923 Endo H 918:923 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 61 theme Endo 918:921 arg1 H 915:915 endoglycosidase H 899:915 endoglycosidase H (Endo H) 899:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 9 62 theme mammalian 1816:1824 arg1 hosts 1826:1830 mammalian hosts 1816:1830 mammalian hosts 1816:1830 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 4 63 theme viral 736:740 arg1 progeny 742:748 tick cell-derived viral progeny 718:748 tick cell-derived viral progeny 718:748 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 3 64 theme IRE/CTVM20 458:467 arg1 lines 474:478 The IRE/CTVM19 and IRE/CTVM20 cell lines 439:478 lines 474:478 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 15 65 theme molecular 2547:2555 arg1 aspects 2557:2563 molecular aspects 2547:2563 molecular aspects of tick-derived viral particles 2547:2595 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 13 66 theme high 2342:2345 arg1 titers 2347:2352 high titers 2342:2352 high titers 2342:2352 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 10 67 theme growing 1881:1887 arg1 threat 1889:1894 a growing threat 1879:1894 a growing threat to humans 1879:1904 IMPORTANCE Tick-borne phleboviruses represent a growing threat to humans globally. 27194760 8 68 theme mammalian 1614:1622 arg1 hosts 1624:1628 mammalian hosts 1614:1628 mammalian hosts 1614:1628 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 1 69 theme different 209:217 arg1 continents 219:228 different continents 209:228 different continents 209:228 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 7 70 theme higher 1390:1395 arg1 infectivity 1397:1407 a higher infectivity 1388:1407 a higher infectivity for tick cell-derived viruses 1388:1437 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 6 71 theme GN 1008:1009 arg1 weight 998:1003 the apparent molecular weight 975:1003 the apparent molecular weight of GN 975:1009 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 11 72 theme previous 1986:1993 arg1 studies 1995:2001 previous studies 1986:2001 previous studies 1986:2001 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 14 73 theme specific 2425:2432 arg1 properties 2445:2454 specific structural properties 2425:2454 specific structural properties 2425:2454 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 7 74 theme BHK-21 1363:1368 arg1 cells 1370:1374 tick or BHK-21 cells 1355:1374 cells 1370:1374 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 11 75 theme diseases 1976:1983 arg1 ticks 1925:1929 ticks 1925:1929 ticks 1925:1929 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 11 75 theme diseases 1976:1983 arg1 vectors 1945:1951 important vectors 1935:1951 important vectors of infectious emerging diseases 1935:1983 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 7 76 theme tick 1355:1358 arg1 cells 1370:1374 tick or BHK-21 cells 1355:1374 cells 1370:1374 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 2 77 theme tick 423:426 arg1 reservoir 428:436 the Ixodes ricinus tick reservoir 404:436 the Ixodes ricinus tick reservoir 404:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 12 78 theme host 2118:2121 arg1 alternation 2123:2133 host alternation 2118:2133 host alternation 2118:2133 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 16 79 theme other 2851:2855 arg1 pathogens 2857:2865 other pathogens 2851:2865 other pathogens 2851:2865 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 6 80 from impact 968:973 arg1 viruses 1014:1020 viruses 1014:1020 viruses originating from mammalian BHK-21 cells 1014:1060 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 16 81 from research 2780:2787 arg1 pathogens 2857:2865 other pathogens 2851:2865 other pathogens 2851:2865 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 16 81 from research 2780:2787 arg1 phleboviruses 2812:2824 tick-borne phleboviruses 2801:2824 not only tick-borne phleboviruses but also all viruses 2792:2845 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 16 81 from research 2780:2787 arg1 viruses 2839:2845 all viruses 2835:2845 not only tick-borne phleboviruses but also all viruses 2792:2845 The information gained here lays the basis for future research on not only tick-borne phleboviruses but also all viruses and other pathogens transmitted by ticks. 27194760 5 82 theme envelope 762:769 arg1 protein 777:783 The second envelope viral protein 751:783 The second envelope viral protein 751:783 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 82 theme envelope 762:769 arg1 GN 786:787 GN 786:787 GN 786:787 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 11 83 theme mammalian 2049:2057 arg1 cells 2059:2063 mammalian cells 2049:2063 mammalian cells 2049:2063 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 15 84 theme particles 2587:2595 arg1 aspects 2557:2563 molecular aspects 2547:2563 molecular aspects of tick-derived viral particles 2547:2595 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 13 85 theme tick-borne 2312:2321 arg1 virus 2333:2337 the tick-borne Uukuniemi virus 2308:2337 the tick-borne Uukuniemi virus 2308:2337 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 1 86 theme pathogenic 93:102 arg1 phleboviruses 104:116 novel tick-borne pathogenic phleboviruses 76:116 novel tick-borne pathogenic phleboviruses in the family Bunyaviridae 76:143 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 1 86 theme pathogenic 93:102 arg1 all 146:148 all 146:148 all 146:148 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 6 87 theme electrophoretic 1129:1143 arg1 mobility 1145:1152 the electrophoretic mobility 1125:1152 the electrophoretic mobility of the protein 1125:1167 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 6 87 theme electrophoretic 1129:1143 arg1 slower 1187:1192 slower 1187:1192 slower 1187:1192 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 6 88 theme bonds 1227:1231 arg1 reduction 1204:1212 the reduction 1200:1212 the reduction of disulfide bonds 1200:1231 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 11 89 theme infectious 1956:1965 arg1 diseases 1976:1983 infectious emerging diseases 1956:1983 infectious emerging diseases 1956:1983 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 8 90 attach derived 1491:1497 arg1 cells 1516:1520 vector tick cells 1504:1520 vector tick cells 1504:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 8 90 attach derived 1491:1497 arg2 particles 1481:1489 UUKV particles 1476:1489 UUKV particles derived from vector tick cells 1476:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 7 91 theme focus 1285:1289 arg1 unit 1299:1302 focus forming unit 1285:1302 focus forming unit 1285:1302 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 13 92 theme cell-based 2265:2274 arg1 model 2276:2280 an in vitro tick cell-based model 2248:2280 an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers 2248:2352 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 1 93 theme family 125:130 arg1 Bunyaviridae 132:143 the family Bunyaviridae 121:143 the family Bunyaviridae 121:143 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 7 94 theme nucleoprotein 1267:1279 arg1 amount 1251:1256 the amount 1247:1256 the amount of viral nucleoprotein per focus forming unit 1247:1302 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 7 94 theme nucleoprotein 1267:1279 arg1 nucleoprotein 1267:1279 viral nucleoprotein 1261:1279 viral nucleoprotein 1261:1279 Furthermore, the amount of viral nucleoprotein per focus forming unit differed markedly whether viruses were produced in tick or BHK-21 cells, suggesting a higher infectivity for tick cell-derived viruses. 27194760 15 95 theme host 2652:2655 arg1 switch 2657:2662 the host switch 2648:2662 the host switch in studying early virus-mammalian receptor/cell interactions 2648:2723 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 9 96 theme arthropod 1714:1722 arg1 cells 1731:1735 arthropod vector cells 1714:1735 arthropod vector cells in investigations of the cell biology of arbovirus transmission 1714:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 13 97 theme virus 2333:2337 arg1 production 2294:2303 production 2294:2303 production of the tick-borne Uukuniemi virus to high titers 2294:2352 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 8 98 from infection 1601:1609 arg1 hosts 1624:1628 mammalian hosts 1614:1628 mammalian hosts 1614:1628 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 4 99 gly glycoprotein 668:679 arg1 glycoprotein 668:679 The glycoprotein GC 664:682 The glycoprotein GC 664:682 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 12 100 from importance 2104:2113 arg1 transmission 2144:2155 virus transmission 2138:2155 virus transmission to humans 2138:2165 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 12 100 from importance 2104:2113 arg1 level 2206:2210 the molecular level 2192:2210 the molecular level 2192:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 12 100 from importance 2104:2113 arg1 infection 2179:2187 initial infection 2171:2187 initial infection at the molecular level 2171:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 5 101 theme endoglycosidase 899:913 arg1 H 923:923 Endo H 918:923 Endo H 918:923 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 101 theme endoglycosidase 899:913 arg1 glycosidases 847:858 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) 833:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 5 101 theme endoglycosidase 899:913 arg1 H 915:915 endoglycosidase H 899:915 endoglycosidase H (Endo H) 899:924 The second envelope viral protein, GN, carried mostly N-glycans not recognized by the classical glycosidases peptide-N-glycosidase F (PNGase F) and endoglycosidase H (Endo H). 27194760 13 102 dep in 2251:2252 arg1 vitro 2254:2258 vitro 2254:2258 vitro 2254:2258 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 2 103 attach isolated 390:397 arg1 reservoir 428:436 the Ixodes ricinus tick reservoir 404:436 the Ixodes ricinus tick reservoir 404:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 2 103 attach isolated 390:397 arg2 virus 384:388 the authentic virus 370:388 the authentic virus isolated from the Ixodes ricinus tick reservoir 370:436 To reproduce the tick-mammal switch in vitro, we first established a reverse genetics system to rescue UUKV with a genome close to that of the authentic virus isolated from the Ixodes ricinus tick reservoir. 27194760 6 104 theme molecular 988:996 arg1 weight 998:1003 the apparent molecular weight 975:1003 the apparent molecular weight of GN 975:1009 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 8 105 theme UUKV 1476:1479 arg1 particles 1481:1489 UUKV particles 1476:1489 UUKV particles derived from vector tick cells 1476:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 8 106 contain have 1522:1525 arg1 particles 1481:1489 UUKV particles 1476:1489 UUKV particles derived from vector tick cells 1476:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 8 106 contain have 1522:1525 arg2 specificities 1556:1568 structural specificities 1545:1568 structural specificities 1545:1568 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 8 106 contain have 1522:1525 arg2 glycosylation 1527:1539 glycosylation 1527:1539 glycosylation 1527:1539 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 1 107 from phleboviruses 104:116 arg1 Bunyaviridae 132:143 the family Bunyaviridae 121:143 the family Bunyaviridae 121:143 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 1 108 theme novel 76:80 arg1 phleboviruses 104:116 novel tick-borne pathogenic phleboviruses 76:116 novel tick-borne pathogenic phleboviruses in the family Bunyaviridae 76:143 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 1 108 theme novel 76:80 arg1 all 146:148 all 146:148 all 146:148 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 9 109 theme biology 1767:1773 arg1 investigations 1740:1753 investigations 1740:1753 investigations of the cell biology of arbovirus transmission 1740:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 4 110 theme glycoprotein 668:679 arg1 GC 681:682 The glycoprotein GC 664:682 The glycoprotein GC 664:682 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 0 111 theme Virus 32:36 arg1 Model 38:42 a Tick-Borne Virus Model 19:42 a Tick-Borne Virus Model 19:42 Uukuniemi Virus as a Tick-Borne Virus Model. 27194760 0 111 theme Virus 32:36 arg1 Virus 10:14 Virus 10:14 Virus 10:14 Uukuniemi Virus as a Tick-Borne Virus Model. 27194760 14 112 theme mammalian 2509:2517 arg1 cells 2519:2523 mammalian cells 2509:2523 mammalian cells 2509:2523 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 9 113 theme arbovirus 1778:1786 arg1 transmission 1788:1799 arbovirus transmission 1778:1799 arbovirus transmission 1778:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 8 114 theme vector 1504:1509 arg1 cells 1516:1520 vector tick cells 1504:1520 vector tick cells 1504:1520 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 15 115 theme early 2676:2680 arg1 interactions 2712:2723 early virus-mammalian receptor/cell interactions 2676:2723 early virus-mammalian receptor/cell interactions 2676:2723 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 14 116 attach derived 2396:2402 arg2 virions 2388:2394 virions 2388:2394 virions derived from tick cells 2388:2418 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 14 116 attach derived 2396:2402 arg1 cells 2414:2418 tick cells 2409:2418 tick cells 2409:2418 Using this system, we found that virions derived from tick cells have specific structural properties and N-glycans that may enhance virus infectivity for mammalian cells. 27194760 9 117 from cells 1731:1735 arg1 investigations 1740:1753 investigations 1740:1753 investigations of the cell biology of arbovirus transmission 1740:1799 This study also highlights the importance of working with viruses originating from arthropod vector cells in investigations of the cell biology of arbovirus transmission and entry into mammalian hosts. 27194760 3 118 theme virus 599:603 arg1 production 581:590 production 581:590 production of the virus 581:603 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 27194760 12 119 theme initial 2171:2177 arg1 infection 2179:2187 initial infection 2171:2187 initial infection at the molecular level 2171:2210 This limitation tends to minimize the importance of host alternation in virus transmission to humans and initial infection at the molecular level. 27194760 8 120 theme initial 1593:1599 arg1 infection 1601:1609 the initial infection 1589:1609 the initial infection in mammalian hosts 1589:1628 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 13 121 theme Uukuniemi 2323:2331 arg1 virus 2333:2337 the tick-borne Uukuniemi virus 2308:2337 the tick-borne Uukuniemi virus 2308:2337 With this study, we have developed an in vitro tick cell-based model that allows production of the tick-borne Uukuniemi virus to high titers. 27194760 11 122 theme important 1935:1943 arg1 ticks 1925:1929 ticks 1925:1929 ticks 1925:1929 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 11 122 theme important 1935:1943 arg1 vectors 1945:1951 important vectors 1935:1951 important vectors of infectious emerging diseases 1935:1983 Although ticks are important vectors of infectious emerging diseases, previous studies have mainly involved virus stocks produced in mammalian cells. 27194760 6 123 theme mammalian 1039:1047 arg1 cells 1056:1060 mammalian BHK-21 cells 1039:1060 mammalian BHK-21 cells 1039:1060 Treatment with β-mercaptoethanol did not impact the apparent molecular weight of GN On viruses originating from mammalian BHK-21 cells, GN glycosylations were exclusively sensitive to PNGase F, and the electrophoretic mobility of the protein was substantially slower after the reduction of disulfide bonds. 27194760 10 124 theme IMPORTANCE 1833:1842 arg1 phleboviruses 1855:1867 IMPORTANCE Tick-borne phleboviruses 1833:1867 IMPORTANCE Tick-borne phleboviruses 1833:1867 IMPORTANCE Tick-borne phleboviruses represent a growing threat to humans globally. 27194760 1 125 from decade 68:73 arg1 UNLABELLED 45:54 UNLABELLED 45:54 UNLABELLED 45:54 UNLABELLED In the last decade, novel tick-borne pathogenic phleboviruses in the family Bunyaviridae, all closely related to Uukuniemi virus (UUKV), have emerged on different continents. 27194760 8 126 theme structural 1545:1554 arg1 specificities 1556:1568 structural specificities 1545:1568 structural specificities 1545:1568 Together, our results indicate that UUKV particles derived from vector tick cells have glycosylation and structural specificities that may influence the initial infection in mammalian hosts. 27194760 15 127 link tick-derived 2568:2579 arg1 particles 2587:2595 tick-derived viral particles 2568:2595 tick-derived viral particles 2568:2595 By shedding light on molecular aspects of tick-derived viral particles, our data illustrate the importance of considering the host switch in studying early virus-mammalian receptor/cell interactions. 27194760 4 128 theme cell-derived 723:734 arg1 progeny 742:748 tick cell-derived viral progeny 718:748 tick cell-derived viral progeny 718:748 The glycoprotein GC was mainly highly mannosylated on tick cell-derived viral progeny. 27194760 3 129 theme cell 469:472 arg1 lines 474:478 The IRE/CTVM19 and IRE/CTVM20 cell lines 439:478 lines 474:478 The IRE/CTVM19 and IRE/CTVM20 cell lines, both derived from I. ricinus, were susceptible to the virus rescued from plasmid DNAs and supported production of the virus over many weeks, indicating that infection was persistent. 25267679 8 0 theme Further 1255:1261 arg1 screening 1273:1281 Further secondary screening 1255:1281 Further secondary screening which identified 320 unique molecular structures or validated hits 1255:1348 Further secondary screening which identified 320 unique molecular structures or validated hits was performed. 25267679 9 1 theme cell 1698:1701 arg1 membrane 1703:1710 the Gram-negative cell membrane 1680:1710 the Gram-negative cell membrane 1680:1710 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 3 2 theme glycosylation 459:471 arg1 process 438:444 the process 434:444 the process of flagellin glycosylation 434:471 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 9 3 theme % 1550:1550 arg1 IC50s 1579:1583 IC50s 1579:1583 IC50s 1579:1583 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 3 theme % 1550:1550 arg1 concentrations 1563:1576 50% inhibitory concentrations 1548:1576 50% inhibitory concentrations (IC50s) of approximately 14 μM 1548:1607 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 5 4 theme in 881:882 arg1 [VS 910:912 virtual screening [VS 892:912 virtual screening [VS 892:912 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 5 4 theme in 881:882 arg1 approaches 916:925 in silico (virtual screening [VS]) approaches 881:925 in silico (virtual screening [VS]) approaches 881:925 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 1 5 theme critical 229:236 arg1 role 238:241 a critical role 227:241 a critical role 227:241 Helicobacter pylori is motile by means of polar flagella, and this motility has been shown to play a critical role in pathogenicity. 25267679 0 6 theme key 96:98 arg1 factor 120:125 a key bacterial virulence factor 94:125 a key bacterial virulence factor 94:125 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 9 7 theme μM 1606:1607 arg1 IC50s 1579:1583 IC50s 1579:1583 IC50s 1579:1583 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 7 theme μM 1606:1607 arg1 concentrations 1563:1576 50% inhibitory concentrations 1548:1576 50% inhibitory concentrations (IC50s) of approximately 14 μM 1548:1607 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 8 theme structure-activity 1395:1412 arg1 relationship 1414:1425 structure-activity relationship 1395:1425 structure-activity relationship (SAR) 1395:1431 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 8 theme structure-activity 1395:1412 arg1 SAR 1428:1430 SAR 1428:1430 SAR 1428:1430 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 2 9 theme major 265:269 arg1 proteins 292:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 3 10 theme consequent 522:531 arg1 motility 533:540 consequent motility 522:540 consequent motility 522:540 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 9 11 theme kinetic 1375:1381 arg1 studies 1383:1389 Following kinetic studies 1365:1389 Following kinetic studies 1365:1389 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 7 12 theme reasonable 1198:1207 arg1 inhibition 1209:1218 reasonable inhibition 1198:1218 reasonable inhibition 1198:1218 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 8 13 theme secondary 1263:1271 arg1 screening 1273:1281 Further secondary screening 1255:1281 Further secondary screening which identified 320 unique molecular structures or validated hits 1255:1348 Further secondary screening which identified 320 unique molecular structures or validated hits was performed. 25267679 0 14 theme virulence 110:118 arg1 factor 120:125 a key bacterial virulence factor 94:125 a key bacterial virulence factor 94:125 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 9 15 theme selected 1445:1452 arg1 inhibitors 1454:1463 selected inhibitors 1445:1463 selected inhibitors from our refined list of 320 compounds 1445:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 5 16 theme small-molecule 808:821 arg1 inhibitors 823:832 small-molecule inhibitors 808:832 small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches 808:925 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 3 17 theme H. 489:490 arg1 assembly 509:516 H. pylori flagellar assembly 489:516 H. pylori flagellar assembly 489:516 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 5 18 theme virtual 892:898 arg1 [VS 910:912 virtual screening [VS 892:912 virtual screening [VS 892:912 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 5 18 theme virtual 892:898 arg1 approaches 916:925 in silico (virtual screening [VS]) approaches 881:925 in silico (virtual screening [VS]) approaches 881:925 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 4 19 theme drug 631:634 arg1 target 636:641 an antivirulence drug target 614:641 an antivirulence drug target 614:641 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 4 19 theme drug 631:634 arg1 potential 601:609 considerable potential 588:609 considerable potential 588:609 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 0 20 theme bacterial 100:108 arg1 factor 120:125 a key bacterial virulence factor 94:125 a key bacterial virulence factor 94:125 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 5 21 theme Pse 779:781 arg1 enzymes 796:802 the five Pse biosynthetic enzymes 770:802 the five Pse biosynthetic enzymes 770:802 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 1 22 theme polar 170:174 arg1 flagella 176:183 polar flagella 170:183 polar flagella 170:183 Helicobacter pylori is motile by means of polar flagella, and this motility has been shown to play a critical role in pathogenicity. 25267679 9 23 theme flagella 1737:1744 arg1 formation 1724:1732 formation 1724:1732 formation of flagella 1724:1744 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 4 24 theme antivirulence 617:629 arg1 target 636:641 an antivirulence drug target 614:641 an antivirulence drug target 614:641 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 4 24 theme antivirulence 617:629 arg1 potential 601:609 considerable potential 588:609 considerable potential 588:609 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 8 25 theme molecular 1311:1319 arg1 structures 1321:1330 320 unique molecular structures 1300:1330 320 unique molecular structures 1300:1330 Further secondary screening which identified 320 unique molecular structures or validated hits was performed. 25267679 0 26 theme Small-molecule 0:13 arg1 inhibitors 15:24 Small-molecule inhibitors 0:24 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 0:126 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 6 27 theme 100,000-compound 936:951 arg1 library 953:959 a 100,000-compound library 934:959 a 100,000-compound library 934:959 Using a 100,000-compound library, 1,773 hits that exhibited a 40% threshold inhibition at a 10 μM concentration were identified by HTS. 25267679 4 28 dep H. 686:687 arg1 persistence 713:723 persistence 713:723 persistence 713:723 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 4 28 dep H. 686:687 arg1 colonization 696:707 colonization 696:707 colonization 696:707 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 4 28 dep H. 686:687 arg1 pylori 689:694 H. pylori colonization and persistence 686:723 H. pylori colonization and persistence in the host 686:735 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 5 29 theme biosynthetic 783:794 arg1 enzymes 796:802 the five Pse biosynthetic enzymes 770:802 the five Pse biosynthetic enzymes 770:802 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 6 30 theme μM 1023:1024 arg1 concentration 1026:1038 a 10 μM concentration 1018:1038 a 10 μM concentration 1018:1038 Using a 100,000-compound library, 1,773 hits that exhibited a 40% threshold inhibition at a 10 μM concentration were identified by HTS. 25267679 9 31 theme distinct 1630:1637 arg1 cluster 1648:1654 a distinct chemical cluster 1628:1654 a distinct chemical cluster 1628:1654 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 32 from list 1482:1485 arg1 analysis 1433:1440 analysis 1433:1440 analysis of selected inhibitors from our refined list of 320 compounds 1433:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 32 from list 1482:1485 arg1 inhibitors 1454:1463 selected inhibitors 1445:1463 selected inhibitors from our refined list of 320 compounds 1445:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 2 33 theme flagellin 282:290 arg1 proteins 292:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 5 34 dep in 881:882 arg1 silico 884:889 silico 884:889 silico 884:889 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 9 35 theme chemical 1639:1646 arg1 cluster 1648:1654 a distinct chemical cluster 1628:1654 a distinct chemical cluster 1628:1654 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 2 36 theme structural 271:280 arg1 proteins 292:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 9 37 theme 320 1490:1492 arg1 compounds 1494:1502 320 compounds 1490:1502 320 compounds 1490:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 4 38 theme considerable 588:599 arg1 target 636:641 an antivirulence drug target 614:641 an antivirulence drug target 614:641 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 4 38 theme considerable 588:599 arg1 potential 601:609 considerable potential 588:609 considerable potential 588:609 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 0 39 theme acid 45:48 arg1 pathway 63:69 the pseudaminic acid biosynthetic pathway 29:69 the pseudaminic acid biosynthetic pathway 29:69 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 9 40 theme inhibitors 1454:1463 arg1 analysis 1433:1440 analysis 1433:1440 analysis of selected inhibitors from our refined list of 320 compounds 1433:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 2 41 gly glycosylated 323:334 arg1 proteins 292:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins 261:299 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 9 42 theme 50 1548:1549 arg1 % 1550:1550 % 1550:1550 % 1550:1550 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 6 43 theme threshold 994:1002 arg1 inhibition 1004:1013 a 40% threshold inhibition 988:1013 a 40% threshold inhibition 988:1013 Using a 100,000-compound library, 1,773 hits that exhibited a 40% threshold inhibition at a 10 μM concentration were identified by HTS. 25267679 0 44 theme pseudaminic 33:43 arg1 pathway 63:69 the pseudaminic acid biosynthetic pathway 29:69 the pseudaminic acid biosynthetic pathway 29:69 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 5 45 theme high-throughput 845:859 arg1 HTS 872:874 HTS 872:874 HTS 872:874 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 5 45 theme high-throughput 845:859 arg1 screening 861:869 high-throughput screening 845:869 high-throughput screening (HTS) 845:875 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 9 46 theme Following 1365:1373 arg1 studies 1383:1389 Following kinetic studies 1365:1389 Following kinetic studies 1365:1389 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 7 47 theme 1.6-million 1096:1106 arg1 library 1117:1123 a 1.6-million compound library 1094:1123 a 1.6-million compound library directed at two pathway enzymes 1094:1155 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 9 48 from analysis 1433:1440 arg1 list 1482:1485 our refined list 1470:1485 our refined list of 320 compounds 1470:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 6 49 theme % 992:992 arg1 inhibition 1004:1013 a 40% threshold inhibition 988:1013 a 40% threshold inhibition 988:1013 Using a 100,000-compound library, 1,773 hits that exhibited a 40% threshold inhibition at a 10 μM concentration were identified by HTS. 25267679 0 50 theme pathway 63:69 arg1 inhibitors 15:24 Small-molecule inhibitors 0:24 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 0:126 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 7 51 theme pathway 1141:1147 arg1 enzymes 1149:1155 two pathway enzymes 1137:1155 two pathway enzymes 1137:1155 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 9 52 with inhibitors 1532:1541 arg1 IC50s 1579:1583 IC50s 1579:1583 IC50s 1579:1583 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 52 with inhibitors 1532:1541 arg1 concentrations 1563:1576 50% inhibitory concentrations 1548:1576 50% inhibitory concentrations (IC50s) of approximately 14 μM 1548:1607 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 53 theme inhibitory 1552:1561 arg1 IC50s 1579:1583 IC50s 1579:1583 IC50s 1579:1583 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 9 53 theme inhibitory 1552:1561 arg1 concentrations 1563:1576 50% inhibitory concentrations 1548:1576 50% inhibitory concentrations (IC50s) of approximately 14 μM 1548:1607 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 8 54 theme unique 1304:1309 arg1 structures 1321:1330 320 unique molecular structures 1300:1330 320 unique molecular structures 1300:1330 Further secondary screening which identified 320 unique molecular structures or validated hits was performed. 25267679 3 55 dep H. 489:490 arg1 pylori 492:497 H. pylori 489:497 H. pylori flagellar assembly 489:516 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 0 56 theme biosynthetic 50:61 arg1 pathway 63:69 the pseudaminic acid biosynthetic pathway 29:69 the pseudaminic acid biosynthetic pathway 29:69 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 7 57 theme hits 1171:1174 arg1 4 1177:1177 4 1177:1177 4 1177:1177 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 7 57 theme hits 1171:1174 arg1 hits 1171:1174 80 hits 1168:1174 80 hits 1168:1174 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 9 58 theme compounds 1494:1502 arg1 list 1482:1485 our refined list 1470:1485 our refined list of 320 compounds 1470:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 3 59 theme flagellin 449:457 arg1 glycosylation 459:471 flagellin glycosylation 449:471 flagellin glycosylation 449:471 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 9 60 dep studies 1383:1389 arg1 analysis 1433:1440 analysis 1433:1440 analysis of selected inhibitors from our refined list of 320 compounds 1433:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 8 61 theme validated 1335:1343 arg1 hits 1345:1348 validated hits 1335:1348 validated hits 1335:1348 Further secondary screening which identified 320 unique molecular structures or validated hits was performed. 25267679 1 62 theme Helicobacter 128:139 arg1 pylori 141:146 Helicobacter pylori 128:146 Helicobacter pylori 128:146 Helicobacter pylori is motile by means of polar flagella, and this motility has been shown to play a critical role in pathogenicity. 25267679 4 63 theme biosynthetic 560:571 arg1 pathway 573:579 the Pse biosynthetic pathway 552:579 the Pse biosynthetic pathway 552:579 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 3 64 theme flagellar 499:507 arg1 assembly 509:516 H. pylori flagellar assembly 489:516 H. pylori flagellar assembly 489:516 This glycan is unique to microorganisms, and the process of flagellin glycosylation is required for H. pylori flagellar assembly and consequent motility. 25267679 5 65 theme screening 900:908 arg1 [VS 910:912 virtual screening [VS 892:912 virtual screening [VS 892:912 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 5 65 theme screening 900:908 arg1 approaches 916:925 in silico (virtual screening [VS]) approaches 881:925 in silico (virtual screening [VS]) approaches 881:925 This report describes screening the five Pse biosynthetic enzymes for small-molecule inhibitors using both high-throughput screening (HTS) and in silico (virtual screening [VS]) approaches. 25267679 4 66 theme Pse 556:558 arg1 pathway 573:579 the Pse biosynthetic pathway 552:579 the Pse biosynthetic pathway 552:579 As such, the Pse biosynthetic pathway offers considerable potential as an antivirulence drug target, especially since motility is required for H. pylori colonization and persistence in the host. 25267679 0 67 theme targeting 72:80 arg1 motility 82:89 targeting motility 72:89 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 0:126 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 7 68 theme μM 1228:1229 arg1 concentration 1231:1243 a 10 μM concentration 1223:1243 a 10 μM concentration 1223:1243 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 0 69 dep inhibitors 15:24 arg1 motility 82:89 targeting motility 72:89 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 0:126 Small-molecule inhibitors of the pseudaminic acid biosynthetic pathway: targeting motility as a key bacterial virulence factor. 25267679 9 70 theme refined 1474:1480 arg1 list 1482:1485 our refined list 1470:1485 our refined list of 320 compounds 1470:1502 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 2 71 theme nonulosonate 345:356 arg1 sugar 358:362 the nonulosonate sugar 341:362 the nonulosonate sugar 341:362 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 2 71 theme nonulosonate 345:356 arg1 acid 377:380 pseudaminic acid 365:380 pseudaminic acid (Pse) 365:386 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 2 72 theme pseudaminic 365:375 arg1 Pse 383:385 Pse 383:385 Pse 383:385 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 2 72 theme pseudaminic 365:375 arg1 sugar 358:362 the nonulosonate sugar 341:362 the nonulosonate sugar 341:362 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 2 72 theme pseudaminic 365:375 arg1 acid 377:380 pseudaminic acid 365:380 pseudaminic acid (Pse) 365:386 The major structural flagellin proteins have been shown to be glycosylated with the nonulosonate sugar, pseudaminic acid (Pse). 25267679 9 73 theme Gram-negative 1684:1696 arg1 membrane 1703:1710 the Gram-negative cell membrane 1680:1710 the Gram-negative cell membrane 1680:1710 Following kinetic studies and structure-activity relationship (SAR) analysis of selected inhibitors from our refined list of 320 compounds, we demonstrated that three inhibitors with 50% inhibitory concentrations (IC50s) of approximately 14 μM, which belonged to a distinct chemical cluster, were able to penetrate the Gram-negative cell membrane and prevent formation of flagella. 25267679 7 74 theme compound 1108:1115 arg1 library 1117:1123 a 1.6-million compound library 1094:1123 a 1.6-million compound library directed at two pathway enzymes 1094:1155 In addition, VS efforts using a 1.6-million compound library directed at two pathway enzymes identified 80 hits, 4 of which exhibited reasonable inhibition at a 10 μM concentration in vitro. 25267679 6 75 theme 40 990:991 arg1 % 992:992 % 992:992 % 992:992 Using a 100,000-compound library, 1,773 hits that exhibited a 40% threshold inhibition at a 10 μM concentration were identified by HTS. 28711789 8 0 from metabolism 1241:1250 arg1 wall 1264:1267 the cell wall 1255:1267 the cell wall 1255:1267 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 6 1 theme wall 848:851 arg1 processes 853:861 cell wall processes 843:861 cell wall processes 843:861 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 8 2 theme drought 1201:1207 arg1 stress 1209:1214 drought stress 1201:1214 drought stress 1201:1214 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 10 3 gly glycoproteins 1468:1480 arg1 glycoproteins 1468:1480 the glycoproteins 1464:1480 the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1464:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 4 theme mechanisms 1604:1613 arg1 understanding 1577:1589 further understanding 1569:1589 further understanding of molecular mechanisms of drought response in this important legume 1569:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 5 5 gly glycoproteins 680:692 arg1 glycoproteins 680:692 Thirty five glycoproteins 668:692 Thirty five glycoproteins 668:692 Thirty five glycoproteins were changed in abundance in leaves of common bean under drought. 28711789 2 6 theme post-translational 250:267 arg1 modification 269:280 very important post-translational modification 235:280 very important post-translational modification that is involved in many physiological functions and biological pathways 235:353 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 1 7 theme abiotic 141:147 arg1 conditions 156:165 the major abiotic stress conditions 131:165 the major abiotic stress conditions limiting crop growth and productivity 131:203 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 8 8 theme biochemical 1229:1239 arg1 metabolism 1241:1250 biochemical metabolism 1229:1250 biochemical metabolism in the cell wall 1229:1267 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 2 9 theme many 302:305 arg1 functions 321:329 many physiological functions 302:329 many physiological functions 302:329 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 1 10 theme conditions 156:165 arg1 one 124:126 one 124:126 one 124:126 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 1 10 theme conditions 156:165 arg1 conditions 156:165 the major abiotic stress conditions 131:165 the major abiotic stress conditions limiting crop growth and productivity 131:203 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 7 11 from role 1055:1058 arg1 modification 1073:1084 cell wall modification 1063:1084 cell wall modification 1063:1084 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 10 12 theme drought 1618:1624 arg1 response 1626:1633 drought response 1618:1633 drought response in this important legume 1618:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 5 13 theme common 733:738 arg1 bean 740:743 common bean 733:743 common bean under drought 733:757 Thirty five glycoproteins were changed in abundance in leaves of common bean under drought. 28711789 1 14 theme crop 176:179 arg1 growth 181:186 crop growth 176:186 crop growth 176:186 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 8 15 theme cell 1259:1262 arg1 wall 1264:1267 the cell wall 1255:1267 the cell wall 1255:1267 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 7 16 theme cell 1063:1066 arg1 modification 1073:1084 cell wall modification 1063:1084 cell wall modification 1063:1084 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 10 17 theme important 1643:1651 arg1 legume 1653:1658 this important legume 1638:1658 this important legume 1638:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 18 gly N-glycoproteins 96:110 arg1 N-glycoproteins 96:110 N-glycoproteins 96:110 N-glycoproteins 96:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 10 19 theme present 1425:1431 arg1 study 1433:1437 The present study 1421:1437 The present study 1421:1437 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 20 theme common 53:58 arg1 bean 60:63 common bean 53:63 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 2 21 theme proteins 223:230 arg1 Glycosylation 206:218 Glycosylation 206:218 Glycosylation of proteins 206:230 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 7 22 theme cell 1018:1021 arg1 metabolism 1028:1037 cell wall metabolism 1018:1037 cell wall metabolism 1018:1037 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 10 23 from level 1539:1543 arg1 stress 1501:1506 drought stress 1493:1506 drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1493:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 23 from level 1539:1543 arg1 bean 1518:1521 common bean 1511:1521 common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1511:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 3 24 from involvement 374:384 arg1 mechanism 412:420 the mechanism 408:420 the mechanism of drought response in leaves of common bean 408:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 0 25 from changes 42:48 arg1 bean 60:63 common bean 53:63 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 10 26 theme common 1511:1516 arg1 bean 1518:1521 common bean 1511:1521 common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1511:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 27 theme case 82:85 arg1 study 87:91 A case study 80:91 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 9 28 theme mannose 1362:1368 arg1 structures 1343:1352 structures 1343:1352 structures of high mannose, complex and hybrid types of N-glycans 1343:1407 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 3 29 gly N-glycoproteins 389:403 arg1 N-glycoproteins 389:403 N-glycoproteins 389:403 N-glycoproteins 389:403 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 30 theme N-glycoproteins 389:403 arg1 involvement 374:384 the involvement 370:384 the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean 370:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 31 theme response 433:440 arg1 mechanism 412:420 the mechanism 408:420 the mechanism of drought response in leaves of common bean 408:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 9 32 theme complex 1371:1377 arg1 structures 1343:1352 structures 1343:1352 structures of high mannose, complex and hybrid types of N-glycans 1343:1407 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 10 33 theme proteome 1530:1537 arg1 important 1555:1563 important 1555:1563 important 1555:1563 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 33 theme proteome 1530:1537 arg1 level 1539:1543 the proteome level 1526:1543 the proteome level 1526:1543 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 34 theme better 10:15 arg1 understanding 17:29 a better understanding 8:29 a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins 8:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 9 35 theme hybrid 1383:1388 arg1 types 1390:1394 hybrid types 1383:1394 hybrid types of N-glycans 1383:1407 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 6 36 theme related 909:915 arg1 proteins 900:907 proteins 900:907 proteins related to proteolysis 900:930 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 3 37 theme common 455:460 arg1 bean 462:465 common bean 455:465 common bean 455:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 8 38 theme general 1135:1141 arg1 concept 1143:1149 the general concept 1131:1149 the general concept of the stress response in plants 1131:1182 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 6 39 theme proteins 782:789 arg1 majority 764:771 The majority 760:771 The majority of these proteins 760:789 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 0 40 theme changes 42:48 arg1 understanding 17:29 a better understanding 8:29 a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins 8:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 6 41 theme related 879:885 arg1 proteins 887:894 defence/stress related proteins 864:894 defence/stress related proteins 864:894 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 3 42 theme proteomic 470:478 arg1 approach 480:487 a proteomic approach 468:487 a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS 468:547 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 8 43 theme stress 1158:1163 arg1 response 1165:1172 the stress response 1154:1172 the stress response 1154:1172 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 10 44 from response 1626:1633 arg1 legume 1653:1658 this important legume 1638:1658 this important legume 1638:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 4 45 theme label 632:636 arg1 approach 658:665 a label free quantification approach 630:665 a label free quantification approach 630:665 Quantification of N-glycoproteins was performed using MaxQuant with a label free quantification approach. 28711789 3 46 theme affinity 502:509 arg1 chromatography 511:524 lectin affinity chromatography 495:524 lectin affinity chromatography 495:524 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 46 theme affinity 502:509 arg1 LC-MS/MS 540:547 LC-MS/MS 540:547 LC-MS/MS 540:547 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 46 theme affinity 502:509 arg1 SDS-PAGE 527:534 SDS-PAGE 527:534 SDS-PAGE 527:534 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 6 47 theme cell 843:846 arg1 processes 853:861 cell wall processes 843:861 cell wall processes 843:861 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 10 48 from legume 1653:1658 arg1 mechanisms 1604:1613 molecular mechanisms 1594:1613 molecular mechanisms of drought response in this important legume 1594:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 49 theme molecular 1594:1602 arg1 mechanisms 1604:1613 molecular mechanisms 1594:1613 molecular mechanisms of drought response in this important legume 1594:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 1 50 theme major 135:139 arg1 conditions 156:165 the major abiotic stress conditions 131:165 the major abiotic stress conditions limiting crop growth and productivity 131:203 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 6 51 theme functional 812:821 arg1 groups 823:828 functional groups 812:828 functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis 812:930 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 2 52 theme physiological 307:319 arg1 functions 321:329 many physiological functions 302:329 many physiological functions 302:329 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 1 53 theme stress 149:154 arg1 conditions 156:165 the major abiotic stress conditions 131:165 the major abiotic stress conditions limiting crop growth and productivity 131:203 Drought is one of the major abiotic stress conditions limiting crop growth and productivity. 28711789 10 54 theme response 1626:1633 arg1 mechanisms 1604:1613 molecular mechanisms 1594:1613 molecular mechanisms of drought response in this important legume 1594:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 55 theme N-glycoproteins 96:110 arg1 study 87:91 A case study 80:91 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 4 56 theme quantification 643:656 arg1 approach 658:665 a label free quantification approach 630:665 a label free quantification approach 630:665 Quantification of N-glycoproteins was performed using MaxQuant with a label free quantification approach. 28711789 2 57 gly Glycosylation 206:218 arg1 proteins 223:230 proteins 223:230 proteins 223:230 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 2 58 theme biological 335:344 arg1 pathways 346:353 biological pathways 335:353 biological pathways 335:353 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 10 59 from stress 1501:1506 arg1 bean 1518:1521 common bean 1511:1521 common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1511:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 59 from stress 1501:1506 arg1 important 1555:1563 important 1555:1563 important 1555:1563 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 59 from stress 1501:1506 arg1 level 1539:1543 the proteome level 1526:1543 the proteome level 1526:1543 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 60 from bean 60:63 arg1 understanding 17:29 a better understanding 8:29 a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins 8:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 7 61 theme drought 1092:1098 arg1 stress 1100:1105 drought stress 1092:1105 drought stress 1092:1105 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 9 62 theme N-glycans 1288:1296 arg1 structures 1274:1283 The structures 1270:1283 The structures of N-glycans 1270:1296 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 7 63 theme wall 1068:1071 arg1 modification 1073:1084 cell wall modification 1063:1084 cell wall modification 1063:1084 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 4 64 dep label 632:636 arg1 free 638:641 free 638:641 free 638:641 Quantification of N-glycoproteins was performed using MaxQuant with a label free quantification approach. 28711789 2 65 theme important 240:248 arg1 modification 269:280 very important post-translational modification 235:280 very important post-translational modification that is involved in many physiological functions and biological pathways 235:353 Glycosylation of proteins is very important post-translational modification that is involved in many physiological functions and biological pathways. 28711789 4 66 gly N-glycoproteins 580:594 arg1 N-glycoproteins 580:594 N-glycoproteins 580:594 N-glycoproteins 580:594 Quantification of N-glycoproteins was performed using MaxQuant with a label free quantification approach. 28711789 8 67 from concept 1143:1149 arg1 plants 1177:1182 plants 1177:1182 plants 1177:1182 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 0 68 from understanding 17:29 arg1 bean 60:63 common bean 53:63 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 3 69 from mechanism 412:420 arg1 leaves 445:450 leaves 445:450 leaves 445:450 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 10 70 theme related 1482:1488 arg1 glycoproteins 1468:1480 the glycoproteins 1464:1480 the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1464:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 7 71 theme wall 1023:1026 arg1 metabolism 1028:1037 cell wall metabolism 1018:1037 cell wall metabolism 1018:1037 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 9 72 theme high 1357:1360 arg1 mannose 1362:1368 high mannose 1357:1368 high mannose 1357:1368 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 10 73 theme drought 1493:1499 arg1 stress 1501:1506 drought stress 1493:1506 drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume 1493:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 0 74 dep bean 60:63 arg1 study 87:91 A case study 80:91 common bean under drought: A case study of N-glycoproteins 53:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 4 75 theme N-glycoproteins 580:594 arg1 Quantification 562:575 Quantification 562:575 Quantification of N-glycoproteins 562:594 Quantification of N-glycoproteins was performed using MaxQuant with a label free quantification approach. 28711789 7 76 from increase 969:976 arg1 abundance 981:989 abundance 981:989 abundance 981:989 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 0 77 theme protein 34:40 arg1 changes 42:48 protein changes 34:48 protein changes in common bean under drought: A case study of N-glycoproteins 34:110 Towards a better understanding of protein changes in common bean under drought: A case study of N-glycoproteins. 28711789 9 78 theme types 1390:1394 arg1 structures 1343:1352 structures 1343:1352 structures of high mannose, complex and hybrid types of N-glycans 1343:1407 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 3 79 theme drought 425:431 arg1 response 433:440 drought response 425:440 drought response in leaves of common bean 425:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 9 80 theme N-glycans 1399:1407 arg1 complex 1371:1377 complex 1371:1377 complex 1371:1377 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 9 80 theme N-glycans 1399:1407 arg1 types 1390:1394 hybrid types 1383:1394 hybrid types of N-glycans 1383:1407 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 9 80 theme N-glycans 1399:1407 arg1 mannose 1362:1368 high mannose 1357:1368 high mannose 1357:1368 The structures of N-glycans were determined manually from spectra, where structures of high mannose, complex and hybrid types of N-glycans were found. 28711789 3 81 theme lectin 495:500 arg1 chromatography 511:524 lectin affinity chromatography 495:524 lectin affinity chromatography 495:524 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 81 theme lectin 495:500 arg1 LC-MS/MS 540:547 LC-MS/MS 540:547 LC-MS/MS 540:547 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 3 81 theme lectin 495:500 arg1 SDS-PAGE 527:534 SDS-PAGE 527:534 SDS-PAGE 527:534 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 7 82 theme highest 961:967 arg1 increase 969:976 the highest increase 957:976 the highest increase in abundance 957:989 Beta-glucosidase showed the highest increase in abundance among proteins involved in cell wall metabolism, suggesting its role in cell wall modification under drought stress. 28711789 8 83 theme response 1165:1172 arg1 concept 1143:1149 the general concept 1131:1149 the general concept of the stress response in plants 1131:1182 These results fit with the general concept of the stress response in plants and suggest that drought stress might affect biochemical metabolism in the cell wall. 28711789 6 84 theme defence/stress 864:877 arg1 proteins 887:894 defence/stress related proteins 864:894 defence/stress related proteins 864:894 The majority of these proteins were classified into functional groups that include cell wall processes, defence/stress related proteins and proteins related to proteolysis. 28711789 3 85 from response 433:440 arg1 leaves 445:450 leaves 445:450 leaves 445:450 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 28711789 10 86 from mechanisms 1604:1613 arg1 legume 1653:1658 this important legume 1638:1658 this important legume 1638:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 10 87 theme further 1569:1575 arg1 understanding 1577:1589 further understanding 1569:1589 further understanding of molecular mechanisms of drought response in this important legume 1569:1658 The present study provided an insight into the glycoproteins related to drought stress in common bean at the proteome level, which is important for further understanding of molecular mechanisms of drought response in this important legume. 28711789 3 88 from leaves 445:450 arg1 mechanism 412:420 the mechanism 408:420 the mechanism of drought response in leaves of common bean 408:465 To understand the involvement of N-glycoproteins in the mechanism of drought response in leaves of common bean, a proteomic approach using lectin affinity chromatography, SDS-PAGE and LC-MS/MS was applied. 24651839 1 0 theme IgA 262:264 arg1 complexes 273:281 IgA immune complexes 262:281 IgA immune complexes 262:281 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 4 1 theme serum 612:616 arg1 IgA1 618:621 serum IgA1 612:621 serum IgA1 from this patient 612:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 10 2 theme IgA1 1623:1626 arg1 formation 1588:1596 the formation 1584:1596 the formation of unusually polymerized IgA1 1584:1626 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 1 3 theme complexes 273:281 arg1 deposition 248:257 the deposition 244:257 the deposition of IgA immune complexes in the glomerular region 244:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 6 4 theme IgA1 925:928 arg1 profiling 912:920 Glycan profiling 905:920 Glycan profiling of IgA1 by the lectin microarray 905:953 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 5 5 theme additional 806:815 arg1 N-glycosylation 817:831 additional N-glycosylation distinctive 806:843 additional N-glycosylation distinctive 806:843 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 7 6 theme glycan 1124:1129 arg1 profile 1131:1137 the distinct glycan profile 1111:1137 the distinct glycan profile 1111:1137 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 0 7 theme IgA 0:2 arg1 nephropathy 4:14 IgA nephropathy 0:14 IgA nephropathy 0:14 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 0 8 theme aberrant 62:69 arg1 N-glycosylation 71:85 aberrant N-glycosylation 62:85 aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease 62:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 4 9 theme mIgA 685:688 arg1 disorder 702:709 mIgA plasma cell disorder 685:709 mIgA plasma cell disorder 685:709 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 10 10 theme divergent 1641:1649 arg1 mechanisms 1651:1660 divergent mechanisms 1641:1660 divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation 1641:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 10 10 theme divergent 1641:1649 arg1 alterations 1692:1702 multiple structural alterations 1672:1702 multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation 1672:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 0 11 with patient 92:98 arg1 disease 142:148 monoclonal immunoglobulin deposition disease 105:148 monoclonal immunoglobulin deposition disease 105:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 10 12 from results 1471:1477 arg1 case 1503:1506 the rare case 1494:1506 the rare case of a patient with monoclonal immunoglobulin deposition disease 1494:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 0 13 theme immunoglobulin 116:129 arg1 disease 142:148 monoclonal immunoglobulin deposition disease 105:148 monoclonal immunoglobulin deposition disease 105:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 6 14 theme Wisteria 986:993 arg1 WFA 1018:1020 WFA 1018:1020 WFA 1018:1020 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 6 14 theme Wisteria 986:993 arg1 agglutinin 1006:1015 Wisteria floribunda agglutinin 986:1015 Wisteria floribunda agglutinin (WFA) 986:1021 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 0 15 from N-glycosylation 71:85 arg1 patient 92:98 a patient 90:98 a patient with monoclonal immunoglobulin deposition disease 90:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 8 16 theme polymerized 1311:1321 arg1 state 1323:1327 This unusually polymerized state 1296:1327 This unusually polymerized state of IgA1 1296:1335 This unusually polymerized state of IgA1 would cause an increase of the binding avidity for lectins. 24651839 10 17 theme structural 1681:1690 arg1 alterations 1692:1702 multiple structural alterations 1672:1702 multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation 1672:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 18 theme IgA1 1089:1092 arg1 conformation 1073:1084 the native conformation 1062:1084 the native conformation of IgA1 1062:1092 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 1 19 theme glomerulonephritis 208:225 arg1 nephropathy 168:178 Immunoglobulin A nephropathy 151:178 Immunoglobulin A nephropathy (IgAN) 151:185 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 19 theme glomerulonephritis 208:225 arg1 form 192:195 a form 190:195 a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region 190:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 20 from deposition 248:257 arg1 region 301:306 the glomerular region 286:306 the glomerular region 286:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 10 21 with patient 1513:1519 arg1 disease 1563:1569 monoclonal immunoglobulin deposition disease 1526:1569 monoclonal immunoglobulin deposition disease 1526:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 10 22 from case 1503:1506 arg1 results 1471:1477 Our results 1467:1477 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease 1467:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 4 23 theme healthy 741:747 arg1 volunteers 749:758 healthy volunteers 741:758 healthy volunteers 741:758 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 4 24 theme cell 697:700 arg1 disorder 702:709 mIgA plasma cell disorder 685:709 mIgA plasma cell disorder 685:709 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 5 25 from N-glycosylation 817:831 arg1 patient 853:859 this patient 848:859 this patient 848:859 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 1 26 theme A 166:166 arg1 IgAN 181:184 IgAN 181:184 IgAN 181:184 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 26 theme A 166:166 arg1 nephropathy 168:178 Immunoglobulin A nephropathy 151:178 Immunoglobulin A nephropathy (IgAN) 151:185 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 26 theme A 166:166 arg1 form 192:195 a form 190:195 a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region 190:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 5 27 theme IgA1 796:799 arg1 polymerization 778:791 unusual polymerization 770:791 unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies 770:902 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 8 28 theme binding 1368:1374 arg1 avidity 1376:1382 the binding avidity 1364:1382 the binding avidity for lectins 1364:1394 This unusually polymerized state of IgA1 would cause an increase of the binding avidity for lectins. 24651839 7 29 theme additional 1239:1248 arg1 N-glycans 1250:1258 additional N-glycans 1239:1258 additional N-glycans on both the heavy and light chains 1239:1293 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 3 30 theme mesangioproliferative 422:442 arg1 glomerulonephritis 444:461 mesangioproliferative glomerulonephritis 422:461 mesangioproliferative glomerulonephritis 422:461 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 10 31 theme rare 1498:1501 arg1 case 1503:1506 the rare case 1494:1506 the rare case of a patient with monoclonal immunoglobulin deposition disease 1494:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 5 32 theme unusual 770:776 arg1 polymerization 778:791 unusual polymerization 770:791 unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies 770:902 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 10 33 theme glycans 1707:1713 arg1 alterations 1692:1702 multiple structural alterations 1672:1702 multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation 1672:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 34 theme glycan 1207:1212 arg1 conformation 1214:1225 the glycan conformation 1203:1225 the glycan conformation detected as additional N-glycans on both the heavy and light chains 1203:1293 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 10 35 theme monoclonal 1526:1535 arg1 disease 1563:1569 monoclonal immunoglobulin deposition disease 1526:1569 monoclonal immunoglobulin deposition disease 1526:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 1 36 theme glomerular 290:299 arg1 region 301:306 the glomerular region 286:306 the glomerular region 286:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 6 37 theme Glycan 905:910 arg1 profiling 912:920 Glycan profiling 905:920 Glycan profiling of IgA1 by the lectin microarray 905:953 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 10 38 theme patient 1513:1519 arg1 case 1503:1506 the rare case 1494:1506 the rare case of a patient with monoclonal immunoglobulin deposition disease 1494:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 39 theme IgA1 1187:1190 arg1 alteration 1173:1182 the conformational alteration 1154:1182 the conformational alteration 1154:1182 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 0 40 theme unusual 26:32 arg1 polymerization 34:47 unusual polymerization 26:47 unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease 26:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 3 41 theme deposition 507:516 arg1 disease 518:524 monoclonal immunoglobulin deposition disease 481:524 monoclonal immunoglobulin deposition disease associated with monoclonal IgA1 481:556 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 5 42 theme distinctive 833:843 arg1 N-glycosylation 817:831 additional N-glycosylation distinctive 806:843 additional N-glycosylation distinctive 806:843 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 10 43 theme deposition 1552:1561 arg1 disease 1563:1569 monoclonal immunoglobulin deposition disease 1526:1569 monoclonal immunoglobulin deposition disease 1526:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 10 44 theme mesangium 1757:1765 arg1 proliferation 1767:1779 mesangium proliferation 1757:1779 mesangium proliferation 1757:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 4 45 from analyses 600:607 arg1 patient 633:639 this patient 628:639 this patient 628:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 4 46 theme IgA1 618:621 arg1 analyses 600:607 the detailed analyses 587:607 the detailed analyses of serum IgA1 from this patient 587:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 3 47 theme monoclonal 542:551 arg1 IgA1 553:556 monoclonal IgA1 542:556 monoclonal IgA1 542:556 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 9 48 theme polymerized 1432:1442 arg1 IgA1 1461:1464 highly polymerized and glycosylated IgA1 1425:1464 highly polymerized and glycosylated IgA1 1425:1464 WFA specifically recognized highly polymerized and glycosylated IgA1. 24651839 0 49 theme IgA1 52:55 arg1 polymerization 34:47 unusual polymerization 26:47 unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease 26:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 2 50 theme multiple 343:350 arg1 mechanisms 352:361 multiple mechanisms 343:361 multiple mechanisms 343:361 The cause of IgAN is unknown, but multiple mechanisms have been suggested. 24651839 9 51 theme glycosylated 1448:1459 arg1 IgA1 1461:1464 highly polymerized and glycosylated IgA1 1425:1464 highly polymerized and glycosylated IgA1 1425:1464 WFA specifically recognized highly polymerized and glycosylated IgA1. 24651839 6 52 theme lectin 937:942 arg1 microarray 944:953 the lectin microarray 933:953 the lectin microarray 933:953 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 4 53 theme detailed 591:598 arg1 analyses 600:607 the detailed analyses 587:607 the detailed analyses of serum IgA1 from this patient 587:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 1 54 theme immune 266:271 arg1 complexes 273:281 IgA immune complexes 262:281 IgA immune complexes 262:281 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 10 55 theme polymerized 1611:1621 arg1 IgA1 1623:1626 unusually polymerized IgA1 1601:1626 unusually polymerized IgA1 1601:1626 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 56 theme distinct 1115:1122 arg1 profile 1131:1137 the distinct glycan profile 1111:1137 the distinct glycan profile 1111:1137 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 5 57 theme known 887:891 arg1 etiologies 893:902 known etiologies 887:902 known etiologies 887:902 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 0 58 theme deposition 131:140 arg1 disease 142:148 monoclonal immunoglobulin deposition disease 105:148 monoclonal immunoglobulin deposition disease 105:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 6 59 theme floribunda 995:1004 arg1 WFA 1018:1020 WFA 1018:1020 WFA 1018:1020 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 6 59 theme floribunda 995:1004 arg1 agglutinin 1006:1015 Wisteria floribunda agglutinin 986:1015 Wisteria floribunda agglutinin (WFA) 986:1021 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 3 60 with patient 468:474 arg1 disease 518:524 monoclonal immunoglobulin deposition disease 481:524 monoclonal immunoglobulin deposition disease associated with monoclonal IgA1 481:556 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 1 61 theme chronic 200:206 arg1 glomerulonephritis 208:225 chronic glomerulonephritis 200:225 chronic glomerulonephritis 200:225 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 10 62 theme multiple 1672:1679 arg1 alterations 1692:1702 multiple structural alterations 1672:1702 multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation 1672:1779 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 63 theme native 1066:1071 arg1 conformation 1073:1084 the native conformation 1062:1084 the native conformation of IgA1 1062:1092 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 6 64 theme intense 967:973 arg1 signal 975:980 an intense signal 964:980 an intense signal for Wisteria floribunda agglutinin (WFA) 964:1021 Glycan profiling of IgA1 by the lectin microarray revealed an intense signal for Wisteria floribunda agglutinin (WFA). 24651839 7 65 from N-glycans 1250:1258 arg1 light 1282:1286 light 1282:1286 light 1282:1286 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 7 65 from N-glycans 1250:1258 arg1 heavy 1272:1276 heavy 1272:1276 heavy 1272:1276 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 4 66 theme renal 719:723 arg1 involvement 725:735 renal involvement 719:735 renal involvement 719:735 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 4 67 with volunteers 749:758 arg1 disorder 702:709 mIgA plasma cell disorder 685:709 mIgA plasma cell disorder 685:709 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 8 68 theme IgA1 1332:1335 arg1 state 1323:1327 This unusually polymerized state 1296:1327 This unusually polymerized state of IgA1 1296:1335 This unusually polymerized state of IgA1 would cause an increase of the binding avidity for lectins. 24651839 3 69 theme monoclonal 481:490 arg1 disease 518:524 monoclonal immunoglobulin deposition disease 481:524 monoclonal immunoglobulin deposition disease associated with monoclonal IgA1 481:556 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 4 70 from patient 633:639 arg1 analyses 600:607 the detailed analyses 587:607 the detailed analyses of serum IgA1 from this patient 587:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 4 70 from patient 633:639 arg1 IgA1 618:621 serum IgA1 612:621 serum IgA1 from this patient 612:639 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 1 71 theme Immunoglobulin 151:164 arg1 IgAN 181:184 IgAN 181:184 IgAN 181:184 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 71 theme Immunoglobulin 151:164 arg1 nephropathy 168:178 Immunoglobulin A nephropathy 151:178 Immunoglobulin A nephropathy (IgAN) 151:185 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 1 71 theme Immunoglobulin 151:164 arg1 form 192:195 a form 190:195 a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region 190:306 Immunoglobulin A nephropathy (IgAN) is a form of chronic glomerulonephritis characterized by the deposition of IgA immune complexes in the glomerular region. 24651839 5 72 with polymerization 778:791 arg1 N-glycosylation 817:831 additional N-glycosylation distinctive 806:843 additional N-glycosylation distinctive 806:843 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 4 73 theme plasma 690:695 arg1 disorder 702:709 mIgA plasma cell disorder 685:709 mIgA plasma cell disorder 685:709 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 5 74 from etiologies 893:902 arg1 different 872:880 different 872:880 different 872:880 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 10 75 theme analysis 1482:1489 arg1 results 1471:1477 Our results 1467:1477 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease 1467:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 3 76 theme immunoglobulin 492:505 arg1 disease 518:524 monoclonal immunoglobulin deposition disease 481:524 monoclonal immunoglobulin deposition disease associated with monoclonal IgA1 481:556 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 0 77 theme monoclonal 105:114 arg1 disease 142:148 monoclonal immunoglobulin deposition disease 105:148 monoclonal immunoglobulin deposition disease 105:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 0 78 gly N-glycosylation 71:85 arg1 patient 92:98 a patient 90:98 a patient with monoclonal immunoglobulin deposition disease 90:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 0 78 gly N-glycosylation 71:85 arg1 polymerization 34:47 unusual polymerization 26:47 unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease 26:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 24651839 8 79 theme avidity 1376:1382 arg1 increase 1352:1359 an increase 1349:1359 an increase of the binding avidity for lectins 1349:1394 This unusually polymerized state of IgA1 would cause an increase of the binding avidity for lectins. 24651839 3 80 theme glomerulonephritis 444:461 arg1 case 414:417 a rare case 407:417 a rare case of mesangioproliferative glomerulonephritis 407:461 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 10 81 from analysis 1482:1489 arg1 case 1503:1506 the rare case 1494:1506 the rare case of a patient with monoclonal immunoglobulin deposition disease 1494:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 7 82 dep heavy 1272:1276 arg1 chains 1288:1293 chains 1288:1293 chains 1288:1293 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 7 82 dep heavy 1272:1276 arg1 the 1268:1270 the 1268:1270 the 1268:1270 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 10 83 theme IgA1 1737:1740 arg1 deposition 1742:1751 IgA1 deposition 1737:1751 IgA1 deposition 1737:1751 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 3 84 theme rare 409:412 arg1 case 414:417 a rare case 407:417 a rare case of mesangioproliferative glomerulonephritis 407:461 We previously reported a rare case of mesangioproliferative glomerulonephritis in a patient with monoclonal immunoglobulin deposition disease associated with monoclonal IgA1. 24651839 4 85 with patients 671:678 arg1 disorder 702:709 mIgA plasma cell disorder 685:709 mIgA plasma cell disorder 685:709 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 7 86 theme conformational 1158:1171 arg1 alteration 1173:1182 the conformational alteration 1154:1182 the conformational alteration 1154:1182 This signal was reduced by disrupting the native conformation of IgA1, suggesting that the distinct glycan profile was reflecting the conformational alteration of IgA1, including the glycan conformation detected as additional N-glycans on both the heavy and light chains. 24651839 10 87 theme immunoglobulin 1537:1550 arg1 disease 1563:1569 monoclonal immunoglobulin deposition disease 1526:1569 monoclonal immunoglobulin deposition disease 1526:1569 Our results of analysis in the rare case of a patient with monoclonal immunoglobulin deposition disease suggest that the formation of unusually polymerized IgA1 is caused by divergent mechanisms including multiple structural alterations of glycans, which contributes to IgA1 deposition and mesangium proliferation. 24651839 4 88 with comparison 644:653 arg1 those 660:664 those 660:664 those 660:664 In this study, we performed the detailed analyses of serum IgA1 from this patient in comparison with those from patients with mIgA plasma cell disorder without renal involvement and healthy volunteers. 24651839 9 89 gly glycosylated 1448:1459 arg1 IgA1 1461:1464 highly polymerized and glycosylated IgA1 1425:1464 highly polymerized and glycosylated IgA1 1425:1464 WFA specifically recognized highly polymerized and glycosylated IgA1. 24651839 2 90 theme IgAN 322:325 arg1 unknown 330:336 unknown 330:336 unknown 330:336 The cause of IgAN is unknown, but multiple mechanisms have been suggested. 24651839 2 90 theme IgAN 322:325 arg1 cause 313:317 The cause 309:317 The cause of IgAN 309:325 The cause of IgAN is unknown, but multiple mechanisms have been suggested. 24651839 5 91 gly N-glycosylation 817:831 arg1 polymerization 778:791 unusual polymerization 770:791 unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies 770:902 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 5 91 gly N-glycosylation 817:831 arg1 patient 853:859 this patient 848:859 this patient 848:859 We found unusual polymerization of IgA1 with additional N-glycosylation distinctive in this patient, which was different from known etiologies. 24651839 0 92 with polymerization 34:47 arg1 N-glycosylation 71:85 aberrant N-glycosylation 62:85 aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease 62:148 IgA nephropathy caused by unusual polymerization of IgA1 with aberrant N-glycosylation in a patient with monoclonal immunoglobulin deposition disease. 25282312 2 0 theme homobifunctional 654:669 arg1 linkers 693:699 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 631:699 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 631:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 1 from reaction 593:600 arg1 reactivity 534:543 the reactivity 530:543 the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 530:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 1 2 theme glycoconjugate 267:280 arg1 vaccine 282:288 a glycoconjugate vaccine 265:288 a glycoconjugate vaccine against Tuberculosis (TB) 265:314 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 2 3 theme 2-iminomethoxyethyl 631:649 arg1 linkers 693:699 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 631:699 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 631:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 7 4 theme glycoconjugates 1898:1912 arg1 preparation 1851:1861 the preparation 1847:1861 the preparation of highly pure and homogeneous neo glycoconjugates 1847:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 4 5 theme porous 1152:1157 arg1 extraction 1190:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 2 6 theme 4-nitrophenyl 672:684 arg1 ester 686:690 4-nitrophenyl ester 672:690 4-nitrophenyl ester 672:690 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 7 7 theme homogeneous 1882:1892 arg1 glycoconjugates 1898:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 4 8 theme carbon 1171:1176 arg1 extraction 1190:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 1 9 theme pilot 212:216 arg1 study 218:222 a pilot study 210:222 a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier 210:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 4 10 theme electrospray 1277:1288 arg1 spectrometry 1295:1306 electrospray mass spectrometry 1277:1306 electrospray mass spectrometry (ESI-MS(n)) 1277:1318 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 4 10 theme electrospray 1277:1288 arg1 ESI-MS 1309:1314 ESI-MS 1309:1314 ESI-MS(n) 1309:1317 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 7 11 theme pure 1873:1876 arg1 glycoconjugates 1898:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 0 12 theme novel 135:139 arg1 glycovaccine 141:152 a novel glycovaccine 133:152 a novel glycovaccine for protection against tuberculosis 133:188 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 1 13 theme selected 400:407 arg1 carrier 417:423 a selected protein carrier 398:423 a selected protein carrier 398:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 2 14 theme acids 566:570 arg1 reactivity 534:543 the reactivity 530:543 the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 530:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 15 theme antigenic 782:790 arg1 protein 792:798 the simplest antigenic protein 769:798 the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB) 769:845 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 15 theme antigenic 782:790 arg1 TB10.4 761:766 TB10.4 761:766 TB10.4 761:766 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 3 16 dep degree 918:923 arg1 composition 951:961 the glycoform composition 937:961 the glycoform composition 937:961 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 3 16 dep degree 918:923 arg1 the 900:902 the 900:902 the 900:902 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 1 17 theme rational 232:239 arg1 design 241:246 the rational design 228:246 the rational design 228:246 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 2 18 dep homobifunctional 654:669 arg1 ester 686:690 4-nitrophenyl ester 672:690 4-nitrophenyl ester 672:690 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 4 19 theme hydrophilic 1215:1225 arg1 chromatography 1246:1259 hydrophilic interaction liquid chromatography 1215:1259 hydrophilic interaction liquid chromatography 1215:1259 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 2 20 from reactivity 534:543 arg1 reaction 593:600 the glycosylation reaction 575:600 the glycosylation reaction with monosaccharides 575:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 4 21 theme liquid 1239:1244 arg1 chromatography 1246:1259 hydrophilic interaction liquid chromatography 1215:1259 hydrophilic interaction liquid chromatography 1215:1259 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 2 22 theme ribonuclease 729:740 arg1 protein 720:726 the model protein 710:726 the model protein 710:726 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 22 theme ribonuclease 729:740 arg1 A 742:742 ribonuclease A 729:742 ribonuclease A (RNase A) 729:752 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 22 theme ribonuclease 729:740 arg1 A 751:751 RNase A 745:751 RNase A 745:751 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 5 23 theme different 1336:1344 arg1 specificity 1351:1361 different site specificity 1336:1361 different site specificity 1336:1361 Significantly, different site specificity and glycosylation efficiency were demonstrated for the two linkers, resulting in structurally diverse glycoconjugates. 25282312 1 24 theme well-defined 345:356 arg1 glycans 358:364 well-defined glycans 345:364 well-defined glycans 345:364 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 0 25 theme spectrometry 27:38 arg1 characterization 51:66 Liquid chromatography-mass spectrometry structural characterization 0:66 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins 0:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 7 26 theme experimental 1819:1830 arg1 conditions 1832:1841 suitable experimental conditions 1810:1841 suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates 1810:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 3 27 theme Intact 848:853 arg1 analysis 863:870 Intact protein analysis 848:870 Intact protein analysis 848:870 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 0 28 theme rational 100:107 arg1 design 109:114 the rational design 96:114 the rational design 96:114 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 3 29 theme protein 1091:1097 arg1 residues 1099:1106 protein residues 1091:1106 protein residues 1091:1106 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 2 30 theme surface 552:558 arg1 acids 566:570 the surface amino acids 548:570 the surface amino acids in the glycosylation reaction with monosaccharides 548:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 4 31 theme mass 1290:1293 arg1 spectrometry 1295:1306 electrospray mass spectrometry 1277:1306 electrospray mass spectrometry (ESI-MS(n)) 1277:1318 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 4 31 theme mass 1290:1293 arg1 ESI-MS 1309:1314 ESI-MS 1309:1314 ESI-MS(n) 1309:1317 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 1 32 theme amino 383:387 arg1 acids 389:393 non-antigenic amino acids 369:393 non-antigenic amino acids in a selected protein carrier 369:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 7 33 theme rational 1749:1756 arg1 design 1758:1763 a rational design 1747:1763 a rational design of experiments 1747:1778 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 4 34 theme graphitized 1159:1169 arg1 extraction 1190:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 2 35 theme methods 471:477 arg1 combination 428:438 A combination 426:438 A combination of ESI-MS and LC-MS analytical methods 426:477 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 0 36 theme Liquid 0:5 arg1 spectrometry 27:38 Liquid chromatography-mass spectrometry 0:38 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins 0:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 2 37 theme LC-MS 454:458 arg1 methods 471:477 LC-MS analytical methods 454:477 LC-MS analytical methods 454:477 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 38 theme ESI-MS 443:448 arg1 combination 428:438 A combination 426:438 A combination of ESI-MS and LC-MS analytical methods 426:477 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 1 39 from acids 389:393 arg1 carrier 417:423 a selected protein carrier 398:423 a selected protein carrier 398:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 3 40 theme glycoform 941:949 arg1 composition 951:961 the glycoform composition 937:961 the glycoform composition 937:961 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 5 41 theme glycosylation 1367:1379 arg1 efficiency 1381:1390 glycosylation efficiency 1367:1390 glycosylation efficiency 1367:1390 Significantly, different site specificity and glycosylation efficiency were demonstrated for the two linkers, resulting in structurally diverse glycoconjugates. 25282312 3 42 theme neo 983:985 arg1 glycoconjugates 987:1001 all the prepared neo glycoconjugates 966:1001 all the prepared neo glycoconjugates 966:1001 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 0 43 theme neo 71:73 arg1 glycoproteins 75:87 neo glycoproteins 71:87 neo glycoproteins 71:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 5 44 theme diverse 1457:1463 arg1 glycoconjugates 1465:1479 structurally diverse glycoconjugates 1444:1479 structurally diverse glycoconjugates 1444:1479 Significantly, different site specificity and glycosylation efficiency were demonstrated for the two linkers, resulting in structurally diverse glycoconjugates. 25282312 3 45 theme pronase 1010:1016 arg1 digests 1035:1041 pronase and chymotriptic digests 1010:1041 pronase and chymotriptic digests 1010:1041 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 6 46 theme epitope 1542:1548 arg1 formation 1550:1558 the epitope formation 1538:1558 the epitope formation in TB10.4 1538:1568 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 0 47 gly glycoproteins 75:87 arg1 glycoproteins 75:87 neo glycoproteins 71:87 neo glycoproteins 71:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 3 48 dep map 1060:1062 arg1 reactivity 1077:1086 the reactivity 1073:1086 the reactivity of protein residues 1073:1106 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 6 49 theme acids 1520:1524 arg1 analysis 1498:1505 A computational analysis 1482:1505 A computational analysis of the amino acids involved in the epitope formation in TB10.4 1482:1568 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 1 50 theme vaccine 282:288 arg1 synthesis 252:260 synthesis 252:260 synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier 252:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 1 50 theme vaccine 282:288 arg1 design 241:246 the rational design 228:246 the rational design 228:246 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 3 51 theme chymotriptic 1022:1033 arg1 digests 1035:1041 pronase and chymotriptic digests 1010:1041 pronase and chymotriptic digests 1010:1041 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 2 52 theme simplest 773:780 arg1 protein 792:798 the simplest antigenic protein 769:798 the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB) 769:845 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 52 theme simplest 773:780 arg1 TB10.4 761:766 TB10.4 761:766 TB10.4 761:766 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 7 53 theme conditions 1832:1841 arg1 identification 1792:1805 the identification 1788:1805 the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates 1788:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 4 54 theme solid-phase 1178:1188 arg1 extraction 1190:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 6 55 theme 2-iminomethoxyethyl-saccharide 1594:1623 arg1 activation 1625:1634 2-iminomethoxyethyl-saccharide activation 1594:1634 2-iminomethoxyethyl-saccharide activation 1594:1634 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 0 56 theme glycovaccine 141:152 arg1 synthesis 120:128 synthesis 120:128 synthesis 120:128 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 0 56 theme glycovaccine 141:152 arg1 design 109:114 the rational design 96:114 the rational design 96:114 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 4 57 theme on-line 1144:1150 arg1 extraction 1190:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 on-line porous graphitized carbon solid-phase extraction 1144:1199 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 1 58 theme protein 409:415 arg1 carrier 417:423 a selected protein carrier 398:423 a selected protein carrier 398:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 2 59 theme amino 560:564 arg1 acids 566:570 the surface amino acids 548:570 the surface amino acids in the glycosylation reaction with monosaccharides 548:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 4 60 theme interaction 1227:1237 arg1 chromatography 1246:1259 hydrophilic interaction liquid chromatography 1215:1259 hydrophilic interaction liquid chromatography 1215:1259 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 4 61 gly glycopeptides 1113:1125 arg2 glycopeptides 1113:1125 Neo glycopeptides 1109:1125 Neo glycopeptides 1109:1125 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 6 62 theme selective 1674:1682 arg1 conjugation 1684:1694 a more targeted and selective conjugation 1654:1694 a more targeted and selective conjugation preserving the protein antigenicity 1654:1730 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 2 63 attach isolated 800:807 arg2 TB10.4 761:766 TB10.4 761:766 TB10.4 761:766 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 63 attach isolated 800:807 arg2 protein 792:798 the simplest antigenic protein 769:798 the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB) 769:845 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 63 attach isolated 800:807 arg1 MTB 842:844 MTB 842:844 MTB 842:844 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 63 attach isolated 800:807 arg1 tuberculosis 828:839 Mycobacterium tuberculosis 814:839 Mycobacterium tuberculosis (MTB) 814:845 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 6 64 theme targeted 1661:1668 arg1 conjugation 1684:1694 a more targeted and selective conjugation 1654:1694 a more targeted and selective conjugation preserving the protein antigenicity 1654:1730 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 2 65 theme glycosylation 579:591 arg1 reaction 593:600 the glycosylation reaction 575:600 the glycosylation reaction with monosaccharides 575:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 1 66 theme glycans 358:364 arg1 coupling 333:340 site-specific coupling 319:340 site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier 319:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 3 67 theme glycosylation 904:916 arg1 degree 918:923 glycosylation degree 904:923 glycosylation degree 904:923 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 2 68 theme systematic 499:508 arg1 characterization 510:525 the systematic characterization 495:525 the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 495:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 3 69 theme protein 855:861 arg1 analysis 863:870 Intact protein analysis 848:870 Intact protein analysis 848:870 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 7 70 theme neo 1894:1896 arg1 glycoconjugates 1898:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 highly pure and homogeneous neo glycoconjugates 1866:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 1 71 theme non-antigenic 369:381 arg1 acids 389:393 non-antigenic amino acids 369:393 non-antigenic amino acids in a selected protein carrier 369:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 7 72 theme suitable 1810:1817 arg1 conditions 1832:1841 suitable experimental conditions 1810:1841 suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates 1810:1912 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 2 73 theme reactivity 534:543 arg1 characterization 510:525 the systematic characterization 495:525 the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers 495:699 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 0 74 theme chromatography-mass 7:25 arg1 spectrometry 27:38 Liquid chromatography-mass spectrometry 0:38 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins 0:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 3 75 theme residues 1099:1106 arg1 reactivity 1077:1086 the reactivity 1073:1086 the reactivity of protein residues 1073:1106 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 6 76 theme protein 1711:1717 arg1 antigenicity 1719:1730 the protein antigenicity 1707:1730 the protein antigenicity 1707:1730 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 2 77 theme analytical 460:469 arg1 methods 471:477 LC-MS analytical methods 454:477 LC-MS analytical methods 454:477 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 78 theme model 714:718 arg1 protein 720:726 the model protein 710:726 the model protein 710:726 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 78 theme model 714:718 arg1 A 742:742 ribonuclease A 729:742 ribonuclease A (RNase A) 729:752 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 5 79 theme site 1346:1349 arg1 specificity 1351:1361 different site specificity 1336:1361 different site specificity 1336:1361 Significantly, different site specificity and glycosylation efficiency were demonstrated for the two linkers, resulting in structurally diverse glycoconjugates. 25282312 0 80 theme structural 40:49 arg1 characterization 51:66 Liquid chromatography-mass spectrometry structural characterization 0:66 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins 0:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 7 81 theme experiments 1768:1778 arg1 design 1758:1763 a rational design 1747:1763 a rational design of experiments 1747:1778 Additionally, a rational design of experiments lead to the identification of suitable experimental conditions for the preparation of highly pure and homogeneous neo glycoconjugates. 25282312 6 82 theme computational 1484:1496 arg1 analysis 1498:1505 A computational analysis 1482:1505 A computational analysis of the amino acids involved in the epitope formation in TB10.4 1482:1568 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 6 83 from formation 1550:1558 arg1 TB10.4 1563:1568 TB10.4 1563:1568 TB10.4 1563:1568 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 0 84 theme glycoproteins 75:87 arg1 characterization 51:66 Liquid chromatography-mass spectrometry structural characterization 0:66 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins 0:87 Liquid chromatography-mass spectrometry structural characterization of neo glycoproteins aiding the rational design and synthesis of a novel glycovaccine for protection against tuberculosis. 25282312 1 85 theme site-specific 319:331 arg1 coupling 333:340 site-specific coupling 319:340 site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier 319:423 Hereby we describe a pilot study for the rational design and synthesis of a glycoconjugate vaccine against Tuberculosis (TB) by site-specific coupling of well-defined glycans to non-antigenic amino acids in a selected protein carrier. 25282312 3 86 theme glycoconjugates 987:1001 arg1 profile 929:935 profile 929:935 profile 929:935 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 3 86 theme glycoconjugates 987:1001 arg1 degree 918:923 glycosylation degree 904:923 glycosylation degree 904:923 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 3 87 theme prepared 974:981 arg1 glycoconjugates 987:1001 all the prepared neo glycoconjugates 966:1001 all the prepared neo glycoconjugates 966:1001 Intact protein analysis was carried out to quantify the glycosylation degree and profile the glycoform composition of all the prepared neo glycoconjugates, while pronase and chymotriptic digests were analyzed to map and rank the reactivity of protein residues. 25282312 2 88 from acids 566:570 arg1 reaction 593:600 the glycosylation reaction 575:600 the glycosylation reaction with monosaccharides 575:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 89 theme RNase 745:749 arg1 A 742:742 ribonuclease A 729:742 ribonuclease A (RNase A) 729:752 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 2 89 theme RNase 745:749 arg1 A 751:751 RNase A 745:751 RNase A 745:751 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB). 25282312 4 90 theme Neo 1109:1111 arg1 glycopeptides 1113:1125 Neo glycopeptides 1109:1125 Neo glycopeptides 1109:1125 Neo glycopeptides were purified by on-line porous graphitized carbon solid-phase extraction, separated by hydrophilic interaction liquid chromatography and analyzed by electrospray mass spectrometry (ESI-MS(n)). 25282312 6 91 theme amino 1514:1518 arg1 acids 1520:1524 the amino acids 1510:1524 the amino acids involved in the epitope formation in TB10.4 1510:1568 A computational analysis of the amino acids involved in the epitope formation in TB10.4 addressed the choice to 2-iminomethoxyethyl-saccharide activation, that resulted in a more targeted and selective conjugation preserving the protein antigenicity. 25282312 2 92 with reaction 593:600 arg1 monosaccharides 607:621 monosaccharides 607:621 monosaccharides 607:621 A combination of ESI-MS and LC-MS analytical methods was applied for the systematic characterization of the reactivity of the surface amino acids in the glycosylation reaction with monosaccharides towards 2-iminomethoxyethyl or homobifunctional (4-nitrophenyl ester) linkers, both on the model protein, ribonuclease A (RNase A) and on TB10.4, the simplest antigenic protein isolated from Mycobacterium tuberculosis (MTB).